[Clinical application of a supplement to the extracorporal circulation to produce a pulsatile flow (author's transl)]. Two groups of patients with atherosclerotic coronary artery disease, who underwent aortocoronary bypass operation, were perfused with nonpulsatile flow during extracorporeal circulation (ECC) using membrane oxygenators. One group (MO) was used as a control, while for the other group (PAD) a Pulsatile Assist Device in the arterial line was employed. This apparatus consists of a balloon of 80 ml placed inside a rigid housing. The balloon is compressed by pressurized air or expanded by vacuum supplied by a driving console. The apparatus produced pulse amplitudes between 30 and 50 mm of mercury. Other than a very short-lasting fall in mean arterial pressure, thus showing diminished peripheral resistance, no perceptable advantages were found. Base excess and pH-changes showed no differences, also the given amount of sodium bicarbonate in both groups was the same. On the other hand significantly higher hemolyses took place, increasing with the duration of pulsation. The application of the apparatus as an arterial counterpulsator was possible with limitation in only 5 of 15 patients. In all other patients after a short time massive blood foaming developed in the PAD and the attempts had to be stopped because of the risk of gas embolism. In our opinion this apparatus is an unnecessary supplement to the ECC and as an arterial counterpulsator it seems too dangerous.


Regulation of the induction of alkaline phosphatase in choriocarcinoma cells by sodium butyrate. Alkaline phosphatase is induced in human choriocarcinoma cells by short-chain fatty acids, especially sodium butyrate. This fatty acid increases the phosphatase activity immediately and in a nearly linear fashion. Only phosphatase with an alkaline pH optimum is induced. Both the induced alkaline phosphatase and the basal enzyme are precipitated by antiserum against term-placental alkaline phosphatase, but the choriocarcinoma phosphatase is less stable to heating than is the term-placental enzyme. The induction of alkaline phosphatase activity requires cellular synthesis of protein, RNA and DNA. The regulation of induction probably occurs at the transcriptional level.


Modification of response of mouse neuroblastoma cells in culture by serum type. The effect of various types of serum on morphological and biochemical changes in mouse neuroblastoma cells (clone NBP2) in culture was studied. The extent of spontaneous morphological differentiation varied markedly depending upon the type of serum and was maximal in agammaglobulin calf serum (CS). The extent of morphological differentiation after treatment of cells with cyclic AMP-stimulating agents was also dependent upon serum type and was least pronounced in fetal calf serum. The doubling time and extent of clumping varied with the type of serum. The activity of tyrosine hydroxylase (TH) in NB cells was dependent upon serum type and it was highest in newborn CS and agammaglobulin CS. Although elevation of intracellular levels of cyclic AMP in NBP2 clone invariably stimulates neurite formation and TH activity, these functions were increased in certain sera without a significant increase in the cellular cyclic AMP levels. The present study shows that neurite formation, growth rate and TH activity are regulated by more than one mode, one of which is mediated by cyclic AMP. The above changes are independently regulated in the sense that the expression of one can be increased in the absence of others.


Rabbit anti-EL4 serum. A reagent which discriminates between murine cytotoxic and suppressor cells. Antisera against the C57B1 (H-2b) mouse lymphoma, EL4, were prepared in rabbits. After absorption with mouse liver, red cells and thymocytes, the antisera appeared to be cytotoxic for a subpopulation of peripheral T cells. The absorbed antisera blocked the immunosuppressive function observed when spleen cells from mice undergoing a graft versus host reaction were added to cells responding in vitro to sheep erythrocytes. This antiserum was unreactive against the cytotoxic cells also induced in this system. These results substantiate our previous finding that these antisera are specific for T-suppressor as opposed to T-helper or cytotoxic lymphocyte subpopulations.


Trypsin and chymotrypsin inhibitor from chick peas. Selective chemical modifications of the inhibitor and isolation of two isoinhibitors. The trypsin and chymotrypsin inhibitor from chick peas (CI) is stable in HCl 0.001 M -- 0.01 M and in KOH 0.01 M -- 0.05 M even after 24 h. Increased KOH concentrations decrease considerably the inhibitory activity already after 1 h. Maleyation and succinylation of the inhibitor resulted in almost full loss of its trypsin-inhibitory activity but had no effect on the chymotrypsin-inhibitory activity. A series of modifications directed towards tyrosyl residues showed that iodination influenced only the chymotrypsin-inhibitory activity; however, nitration and arsanilation affected not only the chymotrypsin-inhibitory activity but also the trypsin-inhibitory activity. Treatment of the inhibitor with CNBr and chloramine T resulted only in a decrease in the chymotrypsin-inhibitory activity indicating that the only methionine is involved in the chymotrypsin-inhibitory activity. When CI-fragment A, previously treated with trypsin at pH 3.75, was further treated with carboxypeptidase B, a release of three lysyl residues per mole protein was found. CI was separated by equilibrium chromatography on SP-Sephadex column into two isoinhibitors, CII and CIII, respectively. Both inhibited trypsin and chymotrypsin with the same specific activity as CI. They differed from each other only in a glutamyl, aspartyl, glycyl and alanyl residue.


A new arylating agent, 2-carboxy-4,6-dinitrochlorobenzene. Reaction with model compounds and bovine pancreatic ribonuclease. The reagent 2-carboxy-4,6-dinitrochlorobenzene (CDNCB) reacts with the imino, amino and sulfhydryl groups of model compounds. At pH 8.2, sulfhydryl groups react much faster than do amines. N alpha-Acetylhistidine, N alpha-acetyltyrosine and N alpha-acetyltryptophan do not react. Poly(L-Lysine) and poly(DL-lysine) react about 50 times as fast as does N alpha-acetyllysine. A dichloroanalog, 6-carboxy-2,4-dinitro-1,3-dichlorobenzene, shows stepwise reactivity with amines. With bovine pancreatic ribonuclease, which contains no sulfhydryl, CDNCB reacts preferentially with the epsilon-amino of Lys-41 at 450 times the rate with the epsilon-amino of N alpha-acetyllysine. The preferential reactivity at Lys-41 is discussed in relation to the pK of Ly-41, the cationic character of the active site cleft, and the mechanism of RNAase action on substrates.


Solid phase synthesis of the protected 27--42 hexadecapeptide of the heavy chain from myeloma immunoglobulin M603. Elimination of side reactions associated with glycyl-2-oxypropionyl-resin. A fully protected 27--42 hexadecapeptide of the variable region of myeloma immunoglobulin M603 was synthesized on a 2-bromopropionyl-resin by the solid phase method. Side reactions due to cyclization of glycyl-2-oxypropionyl-resin were studied under different reaction conditions. The loss of peptide chains at the dipeptide and tripeptide stages due to diketopeperazine formation was also examined. These side reactions were circumvented by using a combination of fragment and stepwise coupling methods. The synthesized protected peptide was removed from the resin in 85% yield by photolysis, and purified by crystallization and by chromatography on a Sephadex LH-60 column.


Studies on trypsin inhibitors. Part IX. Synthesis and trypsin inhibitory activity of the duopentacontapeptide corresponding to the amino acid sequence of porcine pancreatic secretory trypsin inhibitor II (Kazal). The synthesis of the protected duopentacontapeptide corresponding to the entire amino acid sequence I-52 of porcine pancreatic secretory trypsin inhibitor II (Kazal type) is described. The benzyloxycarbonyltetradecapeptide tert-butyloxycarbonylhydrazide (sequence 1-14) was selectively deblocked with trifluoroacetic acid and used to acylate, by the azide procedure, the peptide free base corresponding to the sequence 15-52. The isolated material was purified by ion exchange chromatography and the protecting groups were removed by successive treatments with anhydrous hydrogen fluoride, 1 M piperidine and mercuric acetate. F02M phosphate buffer, pH8. Determination of the inhibitory capacity indicated that the synthetic material is about 50% effective, at 30:1 inhibitor:trypsin molar ratio in inhibiting the tryptic hydrolysis of Nalpha-benzoyl-DL-arginine-4-nitroanilide. Full inhibition was achieved at a higher inhibitor:trypsin molar ratio. The stability constants and the standard free energy of binding of the complex between trypsin and the synthetic inhibitor have been determined.


Effects on learning and memory of 2-week treatments with chlordiazepoxide lactam, N-desmethyldiazepam, oxazepam and methyloxazepam, alone or in combination with alcohol. A double-blind study with 40 healthy students was done in order to measure the effects of a 2-week treatment with chloridiazepoxide lactam (5 mg), nordiazepam (10 mg), oxazepam (15 mg) and methyloxazepam (20 mg) on immediate memory and associative learning. The drugs were administered t.i.d. and the tests were done after the very last capsule was given. It was ingested with a placebo drink and 0.5 g alcohol/kg body weight. Oxazepam and methyloxazepam alone behaved similar to the placebo. Immediate memory was significantly impaired following the treatment with nordiazepam, chlordiazepoxide lactam, alcohol, and after the simultaneous administration of nordiazepam and chlordiazepoxide lactam with alcohol. Chlordiazepoxide lactam was the only drug which alone impaired associative learning. Also alcohol alone, and all the drugs in combination with alcohol retarded learning acquisition.


[Influence of diagnosis, hospital and sex on the effects of bromperidol]. Based on the data analysis of the AMP-findings of 78 schizophrenic patients from 4 clinical trials, the following effects of Bromperidol could be demonstrated: a strong antipsychotic effect with an early onset (after 3 days), little sedation, good improvement of accompanying depressive symptoms, little autonomic but rather strong extrapyramidal side effects. No differences between the effects in men and women were noticed. Differences between the patient populations of two hospitals were small in comparison to the differences between the patients with catatonic and paranoid schizophrenia. In catatonic patients, Bromperidol had a stronger antipsychotic effect and showed an earlier onset of its effects.


Composition of cerebral fluids in goats adapted to high altitude. We explored the ionic composition of cerebral interstitial fluid (cISF) in six unanesthetized goats at sea level (SL) and again after 5 days at a simulated high altitude (HA) of 4,300 m. By measuring net transependymal fluxes of HCO3-, Cl-, and lactate during ventriculocisternal perfusions with lactate-free artificial cerebrospinal fluid (CSF) with various [HCO3-] and [Cl-], we determined [HCO3-] and [Cl-] in the inflowing perfusate that produced zero flux, which are estimates of the concentrations of these ions in cISF. Ventilatory acclimatization to HA was established in the goats with alkaline shift in cisternal CSF pH. At SL zero flux of HCO3- and of Cl- occurred when [HCO3-] and [Cl-] in the perfusate were equal to those in CSF. At HA Cl- flux again was zero when [Cl-] in perfusate and in the goat's own CSF were equal; however, for HCO3-, zero flux occurred at HA when [HCO3-] in perfusate was significantly lower than in CSF. Mean transependymal washout of lactate was 16 times larger at HA than at SL. We conclude that at SL [HCO3-] and [Cl-] in CSF were the same as in cISF. In goats adapted to HA [Cl-] in cISF and in CSF were again equal, whereas [HCO3-] in cISF was lower and [lactate] presumably higher than in CSF. The fluid surrounding the central chemoreceptors appears to be more acidic in goats acclimatized to HA than at SL despite the alkalosis in cisternal CSF. This may contribute to ventilatory acclimatization to HA.


Oxidation of D-thiazolidine-4-carboxylic acid by hog kidney D-aminoacid oxidase. D-Thiazolidine-4-carboxylic acid is a good substrate for hog kidney D-aminoacid oxidase. Data are presented showing that the only oxidation product is delta 3-thiazoline-4-carboxylic acid, which does not undergo further spontaneous degradation. Thus, the oxidation of D-thiazolidine-4-carboxylic acid by D-aminoacid oxidase differs considerably from the oxidation of its L-isomer catalyzed by rat liver mitochondria which gives as final product N-formylcystine, possibly through the intermediate delta 2-thiazoline-4-carboxylic acid.


Main properties of ox kidney uricase.

Main properties of ox kidney uricase. The main properties of ox kidney uricase have been characterized. Optimum pH and temperature, molecular weight, isoelectric point and u.v. absorption spectrum have been determined, and the behaviour with inhibitors, as well as with antibodies, has been studied. The enzyme is very similar to pig liver uricase, so that the possibility of their complete identity must be taken in consideration.


The effect of changes in endolymphatic ion concentrations on the tectorial membrane. The development of a new preparation technique has allowed the effects of fluid-substitution experiments on the tectorial membrane (t.m.) to be studied morphologically and not only, as previously, electrophysiologically. The organ of Corti and t.m. were examined in situ, unfixed and with in vivo-like ionic conditions under light-microscopical control (differential-interference-contrast, magnification 400--800X). An irreversible shrinkage of the marginal and middle zones of the t.m. was observed under the influence of Na+ ions (substitution of endolymph with artificial perilymph). This shrinkage is also seen in specimens prepared for scanning electron microscopy after perfusion of scale media with perilymph. If endolymph is replaced by an isotonic solution of EDTA (ethylenediaminetetraacetate) the t.m. can swell in its vestibulo-tympanal extent, e.g. from 39 to 72 microns. This swelling is reversible on addition of CaCl2. Although the size of the t.m. is not influenced to any great extent by changes in fluid osmolarity, reduction of H+ ion concentration leads to a minimal state of hydration of the protobril system at pH 4.3 (i.e. region of isoelectric point). The mechanism of Na+ and Ca2+ effects are discussed. These results indicate that the subtectorial space is morphologically separated from the scala media proper by the marginal zone.


Separation and partial characterization of two deoxyribonucleic acid polymerases from Spiroplasma citri. The separation and partial characterization of two deoxyribonucleic acid polymerases from Spiroplasma citri have been achieved. The two enzymes had different elution properties on diethylaminoethyl (DEAE) cellulose and differed in their sensitivity to N-ethylmaleimide (NEM), preference for different template-primers, and sedimentation velocity in linear glycerol gradients. The first enzyme activity, ScA, was retained on DEAE-cellulose and was not inhibited by NEM. Activated deoxyribonucleic acid and poly(dA)-oligo(dT12) were the preferred template-primers. Arabinosyl-cytidine triphosphate had no effect. The sedimentation coefficient of ScA was 6.3s. The second activity, ScB, was not retained on DEAE-cellulose and was inhibited by NEM. Poly(dA)-oligo(dT12) was the preferred template-primer, whereas activated DNA was only poorly utilized. ScB was not affected by arabinosyl-cytidine triphosphate, and its sedimentation coefficient was 4.4s. The polymerization activities of the two enzymes were maximum at 37 to 40 degrees C.


Isolation and properties of the protease from the wild-type and mutant strains of Pseudomonas fragi. A simplified procedure for the purification of the extracellular protease of Pseudomonas fragi was developed. The enzyme was isolated from a derepressed mutant producing 40 times the enzyme level of the parental organism. It was collected from culture filtrates by ammonium sulfate precipitation, and it was obtained in pure form by single chromatography on a column of diethylaminoethyl cellulose. The protease had a molecular weight of 52,000 as estimated by sodium dodecyl sulfate-gel electrophoresis and had properties of a classical neutral endopeptidase with the exception of its substrate specificity. Mutants of P. fragi producing proteases of altered substrate specificities were isolated from plates containing elastin as the sole carbon source. The SP-Sephadex elution patterns of enzymes extracted from each mutant examined were complex, suggesting that either the enzyme was autodigested or several active forms could be generated from a common precursor. The substrate specificities of the mutant enzymes were different from that produced by the parental strain.


Allantoate transport in Saccharomyces cerevisiae. Allantoate uptake appears to be mediated by an energy-dependent active transport system with an apparent Michaelis constant of about 50 microM. Cells were able to accumulate allantoate to greater than 3,000 times the extracellular concentration. The rate of accumulation was maximum at pH 5.7 to 5.8. The energy source for allantoate uptake is probably different from that for uptake of the other allantoin pathway intermediates. The latter systems are inhibited by arsenate, fluoride, dinitrophenol, and carboxyl cyanide-m-chlorophenyl hydrazone, whereas allantoate accumulation was sensitive to only dinitrophenol and carboxyl cyanide-m-chlorophenyl hydrazone. Efflux of preloaded allanotate did not occur at detectable levels. However, exchange of intra- and extracellular allantoate was found to occur very slowly. The latter two characteristics are shared with the allantoin uptake system and may result from the sequestering of intracellular allantoate within the cell vacuole. During the course of these studies, we found that, contrary to earlier reports, the reaction catalyzed by allantoinase is freely reversible.


Manganese, an essential trace element for N2 fixation by Rhodospirillum rubrum and Rhodopseudomonas capsulata: role in nitrogenase regulation. Nitrogenase (N(2)ase) from the photosynthetic bacterium Rhodospirillum rubrum can exist in two forms, an unregulated form (N(2)ase A) and a regulatory form (N(2)ase R), the latter being identified in vitro by its need for activation by a Mn(2+)-dependent N(2)ase activating system. The physiological significance of this Mn(2+)-dependent N(2)ase activating system was suggested here by observations that growth of R. rubrum and Rhodopseudomonas capsulata on N(2) gas (a condition that produces active N(2)ase R) required Mn(2+), but growth on ammonia or glutamate did not. Manganese could not be shown to be required for the biosynthesis of either nitrogenase or glutamine synthetase or for glutamine synthetase turnover, but it was required for the in vitro activation of N(2)ases from N(2) and glutamate-grown R. rubrum and R. capsulata cells. Chromatium N(2)ase, in contrast, was always fully active and did not require Mn(2+) activation, suggesting that only the purple nonsulfur bacteria are capable of controlling their N(2)ase activity by this new type of regulatory system. Although R. rubrum could not substitute Fe(2+) for Mn(2+) in the in vivo N(2) fixation process, Fe(2+) and, to a lesser extent, Co(2+) could substitute for Mn(2+) in the in vitro activation of N(2)ase. Electron paramagnetic resonance spectroscopy of buffer-washed R. rubrum chromatophores showed lines characteristic of Mn(2+). Removal of the Mn(2+)-dependent N(2)ase activating factor by a salt wash of the chromatophores removed 90% of the Mn(2+), which suggested a specific coupling of this metal to the activating factor. The data presented here all indicate that Mn(2+) plays an important physiological role in regulating the N(2) fixation process by these photosynthetic bacteria.


Studies on regulatory functions of malic enzymes. VII. Structural and functional characteristics of sulfhydryl groups in NADP-linked malic enzyme from Escherichia coli W. NADP-linked malic enzyme from Escherichia coli W contains 7 cysteinyl residues per enzyme subunit. The reactivity of sulfhydryl (SH) groups of the enzyme was examined using several SH reagents, including 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and N-ethylmaleimide (NEM). 1. Two SH groups in the native enzyme subunit reacted with DTNB (or NEM) with different reaction rates, accompanied by a complete loss of the enzyme activity. The second-order modification rate constant of the "fast SH group" with DTNB coincided with the second-order inactivation rate constant of the enzyme by the reagent, suggesting that modification of the "fast SH group" is responsible for the inactivation. When the enzyme was denatured in 4 M guanidine HCl, all the SH groups reacted with the two reagents. 2. Althoug the inactivation rate constant was increased by the addition of Mg2+, an essential cofactor in the enzyme reaction, the modification rate constant of the "fast SH group" was unaffected. The relationship between the number of SH groups modified with DTNB or NEM and the residual enzyme activity in the absence of Mg2+ was linear, whereas that in the presence of Mg2+ was concave-upwards. These results suggest that the Mg2+-dependent increase in the inactivation rate constant is not the result of an increase in the rate constant of the "fast FH group" modification. 3. The absorption spectrum of the enzyme in the ultraviolet region was changed by addition of Mg2+. The dissociation constant of the Mg2+-enzyme complex obtained from the Mg2+- dependent increment of the difference absorption coincided with that obtained from the Mg2+- dependent enhancement of NEM inactivation. 4. Both the inactivation rate constant and the modification rate constant of the "fast SH group" were decreased by the addition of NADP+. The protective effect of NADP+ was increased by the addition of Mg2+. Based on the above results, the effects of Mg2+ on the SH-group modification are discussed from the viewpoint of conformational alteration of the enzyme.


Phosphoenolpyruvate carboxylase of Escherichia coli. Affinity labeling with bromopyruvate. Phosphoenolpyruvate carboxylase [EC 4.1.1.31] from Escherichia coli W was alkylated by incubation with bromopyruvate, substrate analog, leading to irreversible inactivation. The reaction followed pseudo-first-order kinetics. Mg2+, an essential cofactor for catalysis, enhanced the inactivation, and the enhancing effect increased as the pH increased. The inactivation rate showed a tendency to saturate with increasing concentrations of bromopyruvate, indicating that an enzyme-bromopyruvate complex was formed prior to the alkylation. DL-Phospholactate, a potent competitive inhibitor with respect to phosphoenolpyruvate, protected the enzyme from inactivation in a competitive manner. Examination of the acid hydrolysate of the enzyme modified with [14C]bromopyruvate by paper chromatography showed that radioactivity was solely incorporated into carboxyhydroxyethyl cysteine. In addition, determination of sulfhydryl groups of the native and modified enzymes with 5,5'-dithiobis(2-nitrobenzoate) showed that inactivation occurred concomitant with the modification of one cysteinyl residue per subunit. The results indicate that bromopyruvate reacted with the enzyme as an active-site-directed reagent.


Preparation and characterization of an active lysozyme derivative: Kyn 62-lysozyme. A novel method for the preparation of Kyn 62-lysozyme, in which tryptophan 62 is replaced by kynurenine, is reported. Hen egg-white lysozyme was ozonized in aqueous solution to yield one N'-formylkynurenine residue and deformylated with hydrochloric acid in frozen solution at -10 degrees C. Crude Kyn 62-lysozyme was purified by affinity and Bio Rex 70 chromatography successively. Kyn 62-lysozyme retains affinity for chitin and is essentially an active enzyme with a slightly weakened but distinct catalytic activity. After this modification, the enzyme activity was changed differently depending on the kind of substrate. At the individual optimum pH's, lytic activity was largely retained (80% active), but the catalytic efficiency for hydrolyzing glycol chitin was relatively low (30% active). Lysis of M. lysodeikticus cell suspensions was optimally catalyzed by Kyn 62-lysozyme at pH 6.2 and at 0.088 ionic strength. These values are lower by 1.3 pH unit and 0.04 ionic strength, respectively, than those of intact lysozyme. The optimum pH and ionic strength for the hydrolysis of neutral substrates were scarcely affected. These results suggest the significance of electrostatic interaction in the lysis of lysozyme. Relatively limited loss of activity induced by modification of the 62nd residue, which is thought to participate directly in the binding of the substrate at subsite C, is discussed on the basis of the similarity of side chain structure in tryptophan and kynurenine.


A neutral proteinase of monkey liver microsomes. Solubilization, partial purification, and properties. Conditions for the solubilization of membrane-bound neutral proteinase associated with monkey liver microsomes were investigated. Among the reagents tested, deoxycholate, cholate, and some nonionic detergents, including Triton X-100, with hydrophilic-lipophilic balance values of around 13, were effective. The solubilization profile indicated that the enzyme is bound to the microsomal membranes by strong hydrophobic interaction. The enzyme was partially purified from monkey liver microsomal fraction, previously washed with 1 M KCl and 0.05% sodium dodecyl sulfate, by Triton X-100 extraction, followed by chromatography on columns of hydroxylapatite and Sepharose CL-6B. The apparent molecular weight of the enzyme was estimated to be about 88,000 from the elution position on Sepharose CL-6B column chromatography in the presence of 0.5% sodium cholate. It was optimally active at pH 8.0 with heat-denatured casein as a substrate. It was strongly inhibited by diisopropyl phosphorofluoridate and phenylmethanesulfonyl fluoride, indicating that the enzyme is a serine proteinase. EDTA, EGTA, and chymostatin also inhibited the enzyme strongly. Among urea-denatured protein substrates tested, calf thymus histone was hydrolyzed most rapidly, followed by casein, hemoglobin, and bovine serum albumin, whereas practically no hydrolysis occurred with denatured ovalbumin, fibrinogen, and gamma-globulin as substrates.


Electron microscopic analysis of tropomyosin paracrystals. Negatively stained images of divalent cation-induced tropomyosin paracrystals show polymorphic patterns which are almost bipolar. Although these bipolar forms are naturally due to antiparallel arrays of molecules, the precise molecular arrangements have not been clarified yet except in the case of one type of these polymorphic paracrystals by Stewart and McLachlan [(1976) J. Mol. Biol. 103, 251--269]. In the previous paper we showed that the lead-induced polar paracrystal is a parallel and in-register array of tropomyosin molecules. Moreover, we have made it possible to locate a given residue on the staining pattern. By overlapping two photographic transparencies of the polar paracrystal antiparallel, directly observed images of polymorphic bipolar paracrystals could be synthesized photographically with fidelity. The overlap length between N-terminals of antiparallel pairs of molecules could be easily determined without any assumptions. Next, we considered the stabilizing forces involved in the morphogenesis of such polymorphic paracrystals. The cation-bridged attractive forces already proposed by some groups were insufficient to account for the stability of some specific forms of tropomyosin paracrystals. From the primary amino acid sequence of tropomyosin, we calculated the changes of repulsive forces between the basic residues with changes of molecular overlap length between the N-terminals of antiparallel pairs. By setting the values of charge appropriately, we could account well for the stability of the polymorphic structures observed by electron microscopy.


Enzymatic properties of neuraminidases from Arthrobacter ureafaciens. Neuraminidase I and neuraminidase II from Arthrobacter ureafaciens were characterized. As determined by gel filtration on Ultrogel AcA 44, the molecular weights of neuraminidases I and II were 51,000 and 39,000, respectively. Neuraminidases I and II were similar to each other in their enzymatic properties except for the substrate specificities towards gangliosides and erythrocyte stroma. Their optimal pHs were between 5.0 and 5.5 with N-acetylneuraminosyl-lactose or bovine submaxillary mucin as substrates, but with colominic acid as a substrate, the pH optimum was between 4.3 and 4.5. They were most active around 53 degrees C, were stable between pH 6.0 and 9.0, and were thermostable up to 50 degrees C. They did not require Ca2+ for activity and were not inhibited by EDTA. They were inhibited only slightly or not at all by p-chloromercuribenzoic acid of Hg2+. Both neuraminidases I and II were able to hydrolyze the alpha-ketosidic linkage of N-glycolylneuraminic acid as well as that of N-acetylneuraminic acid, and were able to liberate substantially all of the sialic acid from various kinds of substrates. However, they cleaved only about 50% of the sialic acid from bovine submaxillary mucin. The saponification of bovine submaxillary mucin by mild alkali treatment, on the other hand, resulted in an increased susceptibility to the neuraminidases and brought about the complete liberation of sialic acid. Remarkable differences were observed between neuraminidases I and II as regards substrate specificities on gangliosides; the initial rate of hydrolysis by neuraminidase I was 74 times, and its maximum velocity constant was 91 times those of neuraminidase II. The addition of sodium cholate markedly stimulated the enzymatic hydrolysis of gangliosides, and increased the maximum velocity constant of neuraminidase I twofold and that of neuraminidase II 143-fold. Although neuraminidases I and II were able to hydrolyze (alpha,2-3), (alpha,2-6), and (alpha,2-8) linkages, the initial rate of hydrolysis of N-acetylneuraminosyl-alpha,2-6-lactose was greater than that of the alpha,2-3-isomer.


The contractile basis of ameboid movement. VI. The solation-contraction coupling hypothesis. The contracted pellets derived from a high-speed supernate of Dictyostelium discoideum (S3) were investigated to determine the functional activity associated with this specific subset of the cellular motile apparatus. A partially purified model system of gelation and contraction (S6) was prepared from the contracted pellets, and the presence of calcium- and pH-sensitive gelation and contraction in this model demonstrated that a functional cytoskeletal-contratile complex remained at least partially associated with the actin and myosin during contraction. Semi-quantitative assays of gelation and solation in the myosin-free preparation S6 included measurements of turbidity, relative viscosity, and strain birefringence. The extent of gelation was optimal at pH 6.8 and a free calcium ion concentration of approximately 3.0 x 10(-8) M. Solation was favored when the free calcium ion concentration was greater than 7.6 x 10(-7) M or when the pH was increased or decreased from pH 6.8. Gelation was reversibly inhibited by increasing the free calcium ion concentration to approxomately 4.6 x 10(-6) M at pH 6.8. The solation-gelation process of this model has been interpreted to involve the reversible cross-linking of actin filaments. The addition of purified D. discoideum myosin to S6 served to reconstitute calcium- and pH-regulated contraction. The results from this study indicate that contraction is coupled functionally to the local breakdown (solation) of the gel. Therefore, solation has been identified as a structural requirement for extensive shortening during contraction. We have called this concept the solation-contraction coupling hypothesis. Fractionation of a preparation derived from the contracted pellets yielded a fraction consisting of actin and a 95,000-dalton polypeptide that exhibited calcium-sensitive gelation at 28 degrees C and a fraction composed of actin and 30,000- and 18,000-dalton polypeptides that demonstrated calcium-sensitive genlation at 0 degrees C.


An evaluation of 12 methods for the demonstration of penicillinase. Twelve methods for the demonstration of bacterial penicillinase production by strains of Haemophilus influenzae and Staphylococcus aureus are compared, and their suitability for routine clinical laboratory use is evaluated. The acidometric agar plate method is recommended.


Anatomy and ultrastructure of the axons and terminals of neurons R3-R14 in Aplysia. Using light and electron microscopy and autoradiography, we have traced the axons of neurons R3-R14 in the parietovisceral ganglion (PVG) of Aplysia to terminal fields associated with vascular tissue. The axons are identified by their large size (15-30 micrometer diameter), extensive glial infolding, characteristic dense core vesicles (DCV; approximately 180 nm diameter), and specific, rapid uptake of 3H-glycine. Each neuron in this homogeneous group sends an axon via the branchial nerve to the pericardial region surrounding the junction of the efferent gill vein and the heart. R14 also sends axons to major arteries near the PVG. The R3-R14 axons branch extensively; we estimate that there are at least several hundred per cell. Branches along axons in the branchial nerve exit the nerve, subdivide, and end blindly in the sheath which is bathed by hemolymph. Similar blind endings from R3R14 occur in the sheath of the PVG (Coggeshall, '67). Axonal branches in the pericardial region and the special R14 axons in the arterial walls form both varicose endings near and terminals in contact with vasvular smooth muscle. All R3-R14 endings are free of glia, packed with DCV, show occasional omega-shaped profiles and rapidly take up 3H-glycine. R3-R14 manufacture specific low molecular weight peptides (Gainer and Wollberg, '74), and both the cell bodies (Iliffe et al., '77) and the germinals contain unusually high concentrations of glycine. The presence of peptides as putative neurohormones and sheath endings (neurohormonal release areas) are consistent with R3-R14 being neurosecretory (Coggeshall et al., '66). While glycine could not be a circulating hormone due to its high circulating levels (Iliffe et al., '77), glycine could act as a local chemical messenger between R3-R14 and smooth muscle. The terminal morphology of R3-R14 is consistent with these neurons having both synaptic-type and neurosecretory-type functions.


Reversal of innate aversions: attempts to induce a preference for chili peppers in rats. Although humans frequently develop preferences for innately unpalatable bitter or irritant substances, such preferences are extremely rare in animals. An attempt was made to understand the nature of this difference by systematic experiments with laboratory rats, with chili pepper as the unpalatable substance. In parallel with major aspects of the human experience with chili pepper, rats were exposed to it as a flavoring in all their food for periods up to 11 mo from birth, without significant preference enhancement. Gradual introduction of chili into the diet also had no effect, nor did a series of poisoning and safety experiences designed to teach the rats that only chili-flavored foods were safe to eat. A sequence of seven pairings of chili-flavored diet with prompt recovery from thiamine deficiency did significantly attenuate the innate aversion and may have induced a chili preference in at least one case. Extensive experience with chili did not reliably make rats much less sensitive to its oral effects. The only reliable way to eliminate chili aversion in rats is to destroy their chemical irritant sense, which was accomplished in one group of rats. It is concluded that in contrast to humans, it is extremely difficult to reverse innate aversions in rats.


Low strength anthralin in psoriasis.

Low strength anthralin in psoriasis. Anthralin in low concentrations (0.01%--0.03%) was applied in an ointment form once per day in the hospital to 130 psoriasis patients. The treatment program included also the use of UVB light, emollients and bath oil and an antihistamine. Clearing of psoriasis was achieved in all patients in an average time of 4 weeks. The involution of the lesions was carefully studied by correlated clinical observations, light microscopy, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Striking healing of the holes in the stratum corneum and of the gaps in the basement membrane occurred. Low strength anthralin, in addition to its effectiveness, did not produce irritation, staining or side-effects.


pH gradient elution of human IgG1, IgG2 and IgG4 from protein A-sepharose. Pooled human serum having a normal IgG subclass content was chromatographed on a column of protein A-Sepharose. The immunoglobulins that bound to the column at pH 7.0 were eluted with a pH gradient generated by 3 equal volumes of citrate/phosphate buffer at pH 5.0, 4.5 and 2.2. The elution pattern consisted of two major overlapping peaks centered 0.4 pH units apart in the pH gradient; on average the first peak centered at pH 4.7, and the second centered at pH 4.3. Upon second passage of each component, single major peaks centered at the appropriate pH were seen. The subclass distributions of the re-chromatographed peaks were as follows: the high pH-eluting IgG contained less than 1% IgG1, 95% IgG2 and 5% IgG4; the low pH-eluting IgG contained 90% IgG1, 6% Ig2 and 5% IgG4. IgG3 does not bind to protein A and was thus absent from the pH gradient fractions. Chromatography on protein A-Sepharose provides a means for separating normal human IgG1 from IgG2 and may therefore prove useful as an additional tool for studying the relative biological role of these IgG subclasses.


An experimental investigation into the possible origin of pancreatic islet cells from rhombencephalic neurectoderm. To determine whether or not any pancreatic islet cell type arises from rhombencephalic levels of neurectoderm, lengths of presumptive rhombencephalon (containing potential neural crest) of Black Australorp chick embryos at 6- to 9-somite stages were replaced isotopically and isochronically by neural tube of Japanese quail embryos. Some transplants included mesencephalic regions. In some cases various levels of the rhombencephalon were deleted and not replaced. The quail nuclear marker was detected in cranial ganglia in operated embryos sacrificed at 3 3/4 days of incubation and in enteric ganglia and cells accompanying some pancreatic nerves, in embryos killed at 7 days of incubation. This provided evidence of normal migration of crest cells from the grafts. Dopa was administered to the younger embryos, which were submitted to the formaldehyde-induced fluorescence procedure to demonstrate APUD (Amine Precursor Uptake and Decarboxylation) cells. No pancreatic APUD cells exhibited the quail nuclear marker. In 9- to 11-day embryos, A and B cells were identified by specific light and electron microscopic features. None showed the quail marker. The marker was also absent from those D cells seen and from cells of an as yet unidentified type, but not enough of these were found to warrant a conclusion. All islet cell types were found in embryos from which various levels of the rhombencephalon had been deleted. It is concluded that at least A and B islet cells are not derived from the rhombencephalic neurectoderm and probably not from mesencephalic levels. Their most likely origin remains the endoderm, which was the accepted source until recently.


Phenotypically determined resistance of Neisseria gonorrhoeae to normal human serum: environmental factors in subcutaneous chambers in guinea pigs. Some gonococci obtained from human urethral exudate or from subcutaneously implanted chambers in guinea pigs show a resistance to killing by human serum which is lost on sub-culture in vitro after a few generations. The environmental factors which may influence the phenotypic expression of resistance to serum killing were investigated in guinea pig chambers and in chamber fluid in vitro. The redox potential in chambers before and after infection was lower than that of heart blood but conditions were not anaerobic; H2O2 increased the redox potential but did not decrease gonococcal serum resistance. The chambers were slightly alkaline before and after infection. When the concentration of glucose (depleted in infected chambers by the abundant polymorphonuclear cells) was restored to excess, the serum resistance of the gonococci was unaffected. Concentrations of free amino acids in chambers changed little during infection. Gonococci adapted to growth in chambers and subsequently rendered serum-sensitive by growing once on agar reverted to serum-resistance after 0.5 to 1 h incubation in chamber fluid in vitro at 37 degrees C but not at 25 degrees C or 4 degrees C. After 16 to 24 h growth at 37 degrees C, resistance was again lost. The reversion to serum resistance did not occur in a complex laboratory medium. Examination of the chamber fluid after growth of gonococci in vitro showed depletion of lactate, glutamine and proline.


Nigericin-induced death of an acidophilic bacterium. At an external pH of 3.5, nigericin (which catalyses an electroneutral H+/K+ exchange) abolished the transmembrane proton gradient (delta pH) of Bacillus acidocaldarius, causing a rapid acidification of the cytoplasm from approximately pH 6.0 to pH 3.5. A pronounced loss of viability and fine-structural changes rapidly followed treatment with nigericin. A marked decline in respiration and an even more rapid decrease in cytoplasmic ATP were observed. Activity of at least one cytoplasmic enzyme decreased more slowly. There was no generalized loss in the integrity of the cytoplasmic membrane, as assayed by permeability to inulin or Na+ or by release of ultraviolet light-absorbing compounds. The loss of viability upon treatment with carbonyl cyanide m-chlorophenylhydrazone was similar to what observed with nigericin, so proton influx alone, rather than together with K+ efflux, was probably involved in the death of the organism. Moreover, acidification of the cytoplasm rather than abolition of the delta pH was the lethal event, since no loss of viability was observed when the delta pH was abolished by elevation of the external pH.


Cleavage of thiamine by chlorine in tap water. We have clarified that thiamine is cleaved by residual chlorine in proportion to the rise in temperature, pH and concentration of residual chlorine. When rice was boiled in an electric rice cooker, the thiamine in the rice was cleaved by residual chlorine. It is assumed that thiamine is cleaved into hydroxymethylpyrimidine and the thiazole moiety.


Partial purification and properties of proteases from defatted soybean flour. Four chromatographically different proteases were partially purified from defatted soybean flour, and their pH optima were around 5.0 to 5.6 using casein as the substrate. These soybean proteases were designated S1, S2, S3 and S4 according to their order of elution from a DEAE-cellulose column. Each gave a single peak of caseinolytic activity on a Sephadex G-200 column chromatogram, and corresponded to the molecular weights of about 50,000(S1), 35,000(S2), 60,000(S3) and 200,000(S4). The proteases could hydrolyze casein and poly-Glu. alpha-Casein was more rapidly hydrolyzed than beta-casein, but the esters or dipeptide could not be hydrolyzed. Aliquots of 10(-3) M Hg2+, Cu2+ and Zn2+ inhibited the caseinolytic activities by 70% to 90%, while other cations, Mn2+, Mg2+, Ca2+ and Ni2+, at the same concentration did not. SPI (10(-5) M) inhibited 80--90% of their activities, and EPNP (10(-5) M) inhibited their activities 30--60%, but DFP (10(-3) M), SSI (10(-3) M), PCMB (10(-4) M), NEM (10(-3) M) and EDTA (10(-3) M) were not inhibitory. The above results indicate that proteases S1, S2, S3 and S4 from defatted soybean flour can be classified as acid proteases.


Regulation of chloride in quiescent sheep-heart Purkinje fibres studied using intracellular chloride and pH-sensitive micro-electrodes. 1. The intracellular Cl activity, alpha iCl was measured inside quiescent sheep cardiac Purkinje fibres, bathed in normal Tyrode at pH 7.40, buffered with approximately 22 mM-bicarbonate/approximately 5% CO2 + 95% O2. The measurements were made using liquid ion-exchanger Cl-sensitive micro-electrodes. 2. After internal Cl levels had been depleted by prolonged exposure to Cl-free media (glururonate-substituted) when external Cl was restored, there was a rapid re-accumulation of Cl inside the fibres to levels that were much higher than those expected for a passive Cl distribution. Such a process can be conveniently defined as an active inward Cl pump. 3. The inward-pumping was noticeably temperature-sensitive (Q10 approximately 2.6), its rate was reduced about eighteenfold in the nominal absence of external bicarbonate/CO2 and it was substantially inhibited by the drug SITS (4-acetamido-4'-isothiocyanato-stilbene-2,2'-disulphonic acid). 4. The fall of alpha iCl in Cl-free solution was slow and was also equally temperature-sensitive and substantially inhibited by SITS, but was only slightly impaired in the nominal absence of external bicarbonate/CO2. 5. pHi was measured using recessed-tip pH-sensitive micro-electrodes, and in some experiments both pHi and alpha iCl were monitored simultaneously. When alpha iCl slowly declined in Cl-free solution then pHi slowly became alkaline. Upon restoring external Cl, then there was, as usual, a rapid recovery of a high alpha iCl and this was accompanied by a rapid re-acidification of pHi. Both the recovery of alpha iCl and pHi were exponential with virtually the same time constant. 6. Both the slow alkalinization of pHi in Cl-free solution and the rapid re-acidification upon restoring external Cl were substantially inhibited by the drug SITS. 7. When [k]O was raised to 45 mM or more (by removing equivalent amounts of [Na]O), there was a large depolarization of Em and a slow rise of alpha iCl, which was not accompanied by a large change of pHi. The rise of alpha iCl appeared to be unaffected by SITS. 8. It is suggested that a Cl/CHO-3 exchange mechanism can operate reversibly across the membrane of quiescent Purkinje fibres, and that it can account, at least in part, for the high levels of alpha iCl measured in the resting state. It is also concluded that Cl can cross the membrane in other ways, especially in high-K solution possibly by moving passively through conductance channels that are open under these conditions.


Studies on the mechanism of action of acetylcholine antagonists on rat parasympathetic ganglion cells. The mode of action of ACh antagonists on the parasympathetic neurones of the submandibular ganglion of the rat was studied by means of a two-micro-electrode voltage-clamp technique. The currents produced by various agonists (carbachol, ACh, suberylcholine) were studied in steady state and after voltage steps, before and after perfusion of various antagonists. 2. For three antagonists (tubocurarine, hexamethonium, decamethonium) the blocking action increases with hyperpolarization. For three other antagonists (surugatoxin, trimetaphan, mecamylamine) the effects observed at low concentrations appear to be independent of membrane potential, although in some cases voltage dependence of the block was observed for mecamylamine. 3. The blocks the 'open' channel-reception complex. The block produced by tubocurarine, hexamethonium and decamethonium increases with the agonist concentration, an observation which supports a 'sequential' scheme in which the antagonist blocks the 'open' channel-receptor complex. The block produced by trimetaphan and mecamylamine decreases slightly with increased agonist concentration, which in turn suggests that these two compounds are competitive antagonists, preventing binding of the agonists to the closed channel-receptor complex. 4. In the cases where the block is voltage dependent, voltage jumps trigger slow relaxations which are not present in control conditions. In the case of tubocurarine and hexamethonium, the relaxation following a hyperpolarizing voltage jump corresponds to a decrease in conductance. In the case of decamethonium, the slow relaxation is in the opposite direction. 5. The slow relaxations observed with tubocurarine and hexamethonium are speeded by an increase of the antagonist concentration; the slow relaxations observed with decamethonium are slowed by an increase of the decamethonium concentration. 6. The steady-state observations and the relaxations can be interpreted in terms of a scheme in which tubocurarine, hexamethonium and decamethonium act mainly by blocking the channels opened by the cholinergic agonists. 7. The two types of slow relaxation are those predicted if tubocurarine and hexamethonium dissociate slowly from the channel, and decamethonium rapidly. 8. An additional effect of tubocurarine is described, which consists of a potentiation of the rising phase of the response to an ionophoretic pulse. Possible mechanisms of this effect are discussed.


Inter-relationship of sodium, chloride, bicarbonate and acetate transport by the colon of the pig. 1. Net transport of Na, Cl, HCO3 and acetate was examined in the temporarily isolated colon of conscious pigs weighing 46 +/- 8 kg. 2. The entire colon absorbs 4.1 ml. H2O, 0.8 m-equiv Na, 1.3 m-equiv acetate and secretes 0.5 m-equiv HCO3/min with a solution comparable to the normal contents. The absorptive capacity of the proximal and distal halves of the colon was comparable per unit dry weight of mucosa when each segment was presented with the same solution. 3. A series of studies using ion replacement solutions showed that net Na absorption and net HCO3 accumulation in the lumen solution were both increased in the presence of acetate. Cl absorption was independent of Na absorption and was accompanied by an equivalent net secretion of HCO3 in the absence of Na. When NaCl in the perfusion solution was replaced with Na2SO4, Na and HCO3 were absorbed at equal rates. 4. Final pCO2 values observed in NaCl and Na2SO4 solutions were greater than those observed in plasma while the pCO2 of the Na acetate solution after perfusion was reduced to values below plasma concentrations. 5. Results are consistent with the hypothesis that hydration of CO2 in the lumen solution or mucosal cell provides a continuous source of H ions for absorption of the more permeable undissociated acid. The evidence also suggests an additional source of H ions may be provided by a Na-H exchange process located in one of the limiting cell membranes.


Characteristics of saxitoxin binding to the sodium channel of sarcolemma isolated from rat skeletal muscle. 1. The characteristics of saxitoxin (STX) binding to the mammalian Na channel have been studied in purified sarcolemma isolated from rat skeletal muscle. 2. STX binds specifically to isolated sarcolemma with a Kd of 1.43 x 10(-9) M and Bmax of 7-8 p-mole STX bound/mg membrane protein at 0 degrees C in the presence of 140 mM-NaCl. In rat muscle homogenate under the same conditions the corresponding values are Kd = 1.53 x 10(-9) M and Bmax = 0.15-0.20 p-mole/mg protein (18-20 p-mole/g wet wt.). Membrane purification produced a fortyfold increase in STX binding site concentration per milligram protein. Calculated binding site density in isolated sarcolemma was about 30 sites/micron 2 of membrane surface. 3. Denervation (10-14 days) results in a 43% reduction in the density of high-affinity STX binding sites in purified sarcolemma, but the Kd for this class of sites is not changed. 4. In sarcolemma, the apparent Kd for STX binding is dependent on temperature pH and ionic strength. The Q10 for Kd between 0 and 40 degrees C is 1.3. Protonation of a group having a pK of 6.0 markedly raises Kd without affecting Bmax. Apparent Kd increases eightfold when ionic strength is raised from 20 to 600 mM. 5. Dissociation and association rate constants for STX binding are temperature dependent with Q10 of 2.6 and 1.9 respectively between 0 and 20 degrees C. 6. STX binding is competitively inhibited by monovalent and divalent cations under conditions of constant total ionic strength. An affinity sequence of Tl+ greater than Li+ greater than Na+ greater than K+ greater than Rb+ greater than Cs+ is seen for the monovalent cation-binding site. 7. The STX binding site is relatively stable to heat and to enzymic degradation. A specific modifier of carboxyl residues inactivates subsequent STX binding. This process can be prevented by the presence of STX during the reaction. 8. Characteristics of the STX binding site in isolated sarcolemma are compared to those reported for other isolated excitable membranes and for studies of whole muscle and muscle homogenate. Sarcolemma provides a potential source of enriched Na channels for further purification efforts in a mammalian system.


Inhibin activity in ram rete testis fluid: depression of plasma FSH and LH in the castrated and cryptorchid ram. Ram "rete testis" fluid (RTF) routinely collected throughout the year has been used as a source of inhibin. The mean flow rate and mean concentration of spermatozoa in the fluid remained constant during the first 12 days of cannulation. More than 50 castrated or cryptorchid rams have been treated with low doses of steroid-free RTF over a 25-h blood sampling period. Human serum albumin was injected as a control. RTF depressed both FSH and LH plasma levels although the pattern was different for each hormone. There was no change in prolactin secretion. LH secretion was affected first while FSH remained unchanged in castrated and in cryptorchid rams. Thereafter, the maximum depression of FSH plasma levels occurred at a time when LH started to return or had returned to preinjection levels in the cryptorchid and castrated animals respectively. In the cryptorchid rams, RTF suppressed pulsatile LH secretion which was present before treatment but in the castrated animals, RTF lowered LH plasma levels which were constant and showed no pulsatile changes before treatment. Both FSH and LH inhibitory activities have been found in all active fractions obtained by purification of RTF. These activities are papain-sensitive and active fractions have a high apparent molecular weight (greater than or equal to 100 000) as shown by gel filtration and ultrafiltration. These and other results in the literature have lead to a re-definition of inhibin as a protein factor of gonadal origin able to depress plasma levels of FSH and LH, even at low doses.


Biological investigations in alcohol research. The biological mechanisms examined in this paper cover only a small portion of those that may be involved in the pathogenesis of alcoholism. Certain areas on which a great deal of work has already been done have been entirely omitted. Among these are studies on brain acetylcholine and gamma-aminobutyric acid metabolism in alcohol-related conditions; the effect of alcohol on brain proteins and nucleotides and the relationship of changes in these to the development of tolerance and physical dependence; and a variety of other areas involving biochemical and physiological parameters. The omission of any of these areas in no way suggests that they are less significant than those that have been covered. It is just that I have attempted to present a cohesive and coherent review of some areas of biological research which thus far appear to throw light on the clinical development and phenomenology of the alcoholism syndrome. In order to present such a thesis, I have inevitably crowded the evidence to fit some of my pet hypotheses. However, in controversial areas (which are many), I have tried to present some of the evidence on both sides and, hopefully, have succeeded in offering a fair overview of the state of the science as it exists today.


Uridine diphosphate glucose dehydrogenase in normal human synovial cells in culture. Extracts containing uridine diphosphate (UDP) glucose dehydrogenase (EC 1.1.1.22) activity were prepared from 5 normal human synovial cell lines and sources of variation in the method determined. The mean catalytic activity of UDP-glucose dehydrogenase from the 5 extracts was 12.0 +/- 2.4 x 10(-3) International Units/mg protein. The KmUDP-glucose was estimated as 3.90 +/- 1.56 x 10(-5) M and the KmNAD+ was estimated as 1.72 +/- 0.60 x 10(-4) M. Maximum catalytic activity occurred in a temperature range of 55 degrees C-68 degrees C and in a pH range of 8.1-8.4. The mechanistic implications of these data in the normal human diarthrodial joint are discussed.


6-Aryl-4H-s-triazolo[4,3-a][1,4]benzodiazepines. Influence of 1-substitution on pharmacological activity. A series of 1-substituted 6-aryl-4H-s-triazolo[4,3-a][1,4]benzodiazepines was prepared and evaluated for central nervous system activity. It was found that electronegative substituents, such as trifluoromethyl, were detrimental to activity in this series. On the other hand, many compounds with electron-donating substituents at C-1 had interesting activity. In addition to showing anxiolytic potential, some were also active in tests useful for detecting antidepressant and antipsychotic activity. Several analogues with 4-methyl-1-piperazinyl and 4-morpholinyl substituents at C-1 were of particular interest.


Smectite clays in Mars soil: evidence for their presence and role in Viking biology experimental results. Various chemical, physical and geological observations indicate that smectite clays are probably the major components of the Martian soil. Satisfactory ground-based chemical simulation of the Viking biology experimental results was obtained with the smectite clays nontronite and montmorillonite when they contained iron and hydrogen as adsorbed ions. Radioactive gas was released from the medium solution used in the Viking Labeled Release (LR) experiment when interacted with the clays, at rates and quantities similar to those measured by Viking on Mars. Heating of the active clay (mixed with soluble salts) to 160 degrees C in CO2 atmosphere reduced the decomposition activity considerably, again, as was observed on Mars. The decomposition reaction in LR experiment is postulated to be iron-catalyzed formate decomposition on the clay surface. The main features of the Viking Pyrolytic Release (PR) experiment were also simulated recently (Hubbard, 1979) which the iron clays, including a relatively low '1st peak' and significant '2nd peak'. The accumulated observations on various Martian soil properties and the results of simulation experiments, thus indicate that smectite clays are major and active components of the Martian soil. It now appears that many of the results of the Viking biology experiments can be explained on the basis of their surface activity in catalysis and adsorption.


Hypothalamic cathepsin D: assay and isoenzyme composition. A sensitive and convenient method of endopeptidase assay using as substrate globin modified with pyridoxal-5-phosphate was used for determination of acid proteinases in bovine hypothalamus separated by isoelectric focusing. The soluble protein fraction of hypothalamus upon elution from Sephadex gave five peaks of proteinase activity at pH 3.2. The properties indicate that these peaks of endopeptidase activity are isoenzyme forms of cathepsin D.


Effects of cortical ablation on the neurotoxicity and receptor binding of kainic acid in striatum. Lesions of the cerebral cortex alter striatal neuronal vulnerability to locally injected kainic acid. Whereas extensive lesions involving the frontal-parietal-occipital cortex are most effective, lesions limited to the frontal or to the dorsal-lateral parietal cortex offer partial protection. The extensive cortical lesions are associated with selective, marked reductions in the presynaptic markers for glutamatergic afferents in striatum. The protective effects of decortication appear between 6 and 24 hours after the lesion and are maintained up to 30 days after decortication. Whereas decortication results in only a transient reduction of specific receptor binding of [3H]kainic acid to striatal membranes, lesion of striatal intrinsic neurons with kainic acid causes a delayed but marked reduction in specific binding of the ligand. Coadministration of L-glutamic acid (1 mumole) with kainic acid (9 nmoles) partially restores the neurotoxic action of kainic acid in the decorticate striatum; GABA, alanine, and proline (1 mumole) are ineffective with regard to restoring kainate's toxicity for striatal GABAergic neurons. These results suggest that afferent input exerts a permissive effect on the neurotoxic action of kainic acid and that neurotoxicity may involve a cooperative interaction between kainic acid at specific receptors on vulnerable neurons and synaptically released endogenous neurotransmitters, in particular L-glutamic acid.


PCP (phencyclidine): an update.

PCP (phencyclidine): an update. The steady rise in the promiscuous use of phencyclidine (PCP) as a "recreational" drug has recently gained nationwide attention because of the numerous violent and/or bizarre incidents caused by the use of this drug. Because the media often exaggerate reports of bizarre and violent behavior to make a "good" story, the potential PCP user may be tempted to ignore the media warnings. In the case of PCP, however exaggerated the story, a real danger does exist. So, despite numerous newspaper, radio and television warnings about the possible consequences of PCP use and abuse, the incidence of toxic reactions continues to climb. In many cases PCP is sold as other drugs, particularly THC, and in various colored capsules, tablets, liquids and crystals which may explain the increased usage despite the numerous warnings against its use. The advances in laboratory techniques and chemical processess have enabled the clandestine chemist to prepare relatively pure PCP and thus eliminate many of the toxic side effects due to impurities in the drug. In addition, 30 or more psychoactive PCP analogues have been developed and are starting to make an appearance on the street. PCP is perhaps the most potent psychotomimetic compound known at the present time and is capable of inducing a psychosis which is clinically indistinguishable from schizophrenia. The psychosis-producing effects of PCP are the most common toxic effects seen in hospital emergency rooms; but as the amount of PCP taken and/or the simultaneous involvement of other drugs, particularly barbiturates, occurs, severe medical problems (e.g., coma, seizures, respiratory arrest) begin to appear. Death from high doses of PCP or PCP plus other drugs does occur, but the principal cause of death from PCP abuse is due to trauma, homicide or suicide (usually of the bizarre or violent form). Young adult males, persons predisposed to mental illness and naive drug users appear to be the most susceptible to the adverse effects of PCP. The fact that chronic PCP users are starting to increase in number is mute testimony that not all users experience "bad trips" with PCP. Unfortunately for the user, however, this does not guarantee that the next trip will not be a bad one. The effects of chronic use seem to be twofold: severe depression with suicidal thoughts and numerous violent, agitated behavioral patterns. Neither seems to be a suitable alternative. At the present time there is not specific antidote for toxic PCP reactions and the prolonged psychosis induced in some cases does not appear to respond to the standard antipsychotic medications as quickly as do the functional psychoses. The major improvement from a medical standpoint is the development of more sensitive laboratory techniques to confirm the presence of PCP in body fluids. This advance has undoubtedly led to the apparent increase in the number of PCP cases reported by hospitals and to the accuracy of clinical diagnosis by medical, drug or law enforcement communities...


The effect of morphine and some other narcotic analgesics on brain tryptophan concentrations. An acute dose of morphine increased brain tryptophan in mice. This effect was not prevented by naloxone nor was it produced by other narcotic analgesics. Dextrorphan, but not levorphanol, had a similar effect to morphine. A large dose of tryptophan had no effect on the antinociceptive action of morphine in mice. Morphine increased brain tryptophan in rats. This effect was prevented by naloxone. A large dose of tryptophan antagonised the antinociceptive action of morphine in the rat.


Effects of handling stress on plasma enzymes in harp seals, Phoca groenlandica. Three harp seal pups, Phoca groenlandica, were captured on the ice of the Gulf of St. Lawrence, and subjected to 3 h of transportation and handling stress. The activities of creatine kinase (CK), aspartate aminotransferase (AspAT), aldolase, alanine aminotransferase, gamma glutamyl transpeptidase, and leucine aminopeptidase were determined in serial blood samples collected for 4 d following the stress episode. Marked elevation of plasma CK activity was observed 3 h after capture. Values returned to normal in 12 h in two seals, and by 24 h in the third. Slight elevations in AspAT were also noted; the remaining enzymes were unaffected. Plasma CK is recommended as a sensitive indicator of handling stress in seals.


Effects of cadmium on the contractility of a frog cardiac muscle in relation to pH of external solution. In order to examine the pH dependency of Cd effects on cardiac muscle, the electrical and mechanical activities in the bullfrog heart were investigated in relation to various external pHs and concentrations of Cd. The amplitude and duration of the action potential or the spontaneously beating frequency in the atrium were not significantly affected during the 3-min administration of various concentrations of Cd in a range from pH 6 to 10, but the isotonic contraction at the end of the same period was greatly altered: the lower the external pH the more Cd decreased the contractility of the atrium. This pH dependency of Cd effects was also observed in SO4-Ringer's solution whose anions, SO4, were more impermeable than Cl. Decrease in contractility in Cd-Ringer's solution was counteracted by excess Ca. The lower the pH of the Cd-Ringer's solution, the more Ca was necessary to counteract an equal amount of Cd. The amount of Cd-uptake into the atrium was analyzed after soaking the atrium in various concentrations of Cd-Ringer's solution. The higher the concentration of external Cd, the larger the Cd-uptake into the atrium. No pH dependency, however, was observed in this relationship. This suggests that probably only a small fraction of total Cd-uptake interacts with Ca-binding sites which is specific to contraction, and causes the Cd-induced decrease in contractility. This process is considered to be pH-dependent. However, most Cd-uptake into cardiac muscle is pH-independent and may be nonspecific to contraction.


[Etiologic diagnosis of pneumoccocal pneumonia by counterimmunoelectrophoresis (author's transl)]. The antigenic composition of the pneumoccocal capsule and its easy detection by counterimmunoelectrophoresis led us to carry out a comparative study of this method and the results obtained from cultures. A series of 159 patientes with suspected bacterial pneumonia were included in the study. Blood cultures were prepared for all of the patients; sputum cultures were practiced on 35 and cultures of pleural fluid on 32. Serum from all of the patients, urine from 104, sputum from 34, and pleural fluid from 16 were analyzed by counterimmunoelectrophoresis for pneumoccocal antigen. This method was particularly useful for detecting the germ in sputum and pleural fluid. Furthermore, a higher number of positive indexes in serum were obtained by counterimmunoelectrophoresis than by blood cultures (p less than 0.01). This was also true for tests with sputum and pleural fluid. Urine tests resulted in an even greater number of detections. Counterimmunoelectrophoresis is thus a simple, quick method for detecting pneumococcus and is especially useful when accompanied by cultures tests, since they are complementary. We were able to discover a positive indication of penumoccocus in bacterial pneumonia in 39.6 percent of cases with this technique, as opposed to 15 percent using cultures alone.


Characterization of thymidine kinase and phosphorylation of deoxyribonucleosides in Chlamydomonas reinhardti. Using gel filtration chromatography, we find a single peak of deoxythymidine phosphorylating activity in Chlamydomonas reinhardti. This activity has characteristics of a thymidine kinase, in that (1) it will utilize ATP (or dATP) or CTP (or dCTP) as phosphoryl donor, but not AMP or phenyl phosphate, and (2) it is inhibited by dTTP (and less so by dTDP, dUTP, and dUDP) but is unaffected by 3'-5' cyclic AMP. Partially purified chlamydomonas thymidine kinase has a pH optimum near 8.5, and a molecular weight of 80,000 to 85,000 daltons. Kinetic studies indicate a ping-pong mechanism with a Km for thymidine of 1.5 x 10(-7) moles per liter. 5-Bromo- and 5-fluorodeoxyuridine, and to a lesser degree deoxyuridine, are competitive inhibitors, but significant phosphorylation of these nucleotides could not be demonstrated in vitro by thymidine kinase. While thymidine is phosphorylated to dTMP by crude Chlamydomonas extracts, greater than 80% of the product formed by the partially purified enzyme is dTTP. Further, the gel filtration elution position of the single deoxythymidylate kinase activity present in cell extracts coincides with that of thymidine kinase. These results suggest that a multifunctional enzyme, rather than three separate phosphorylating activities, may be responsible for dTTP formation.


Suppressor adherent phagocytic cells in solid tumors: a postulated escape mechanism. A localized graft-versus-host reaction was used to assess the cellular immunity of mice supporting progressive growth of an allogeneic tumor. It was found that early in tumor development anti-tumor-cellular-mediated immunity was hyper-responsive, and that adherent phagocytic cells isolated from the neoplasm would inhibit the specific immunological response. Based on these observations and indirect inference it was postulated that a major factor in the success of an immunogenic tumor is the coterminous development of a normal adherent phagocytic cell population which specifically nullifies, in situ, cellular-mediated anti-tumor immunity.


[Hepatitis-B-surface antigen and panarteritis (author's transl)]. A case of an acutely beginning histologically proved panarteritis is described which was initiated by hepatitis B caused by blood transfusions. After one year of steroid therapy the arteritis was no longer seen histologically, Australia-antigen became negative. Terminally the patient developed an apoplexy, renewed gastric bleeding, septicemia with obstructive jaundice, nose bleeding, increasing renal insufficiency, and cardiac failure. The Australia-antigen reappeared in the serum. It could be assumed that the panarteritis had progressed. Immune complexes of Australia-antigen and corresponding antibodies which are deposited in the vascular wall and cause an inflammatory reaction, are being held responsible for the panateritis. They were proved serologically and by immunofluorescence in the vascular wall. In cases of panarteritis of unknown origin Australia-antigen can be found in a high percentage, as was demonstrated by a second case.


Distribution and metabolic fate of adenosine nucleotides in the membrane of storage vesicles from bovine adrenal medulla. The reactions of adenosine 14C-and gamma 32P-labelled ATP with isolated membranes from catecholamine storage vesicles of the bovine adrenal medulla were studied. In presence of Mg2+ about twice as much of 32P-radioactivity combined with the membrane as 14C-adenosine compounds at 31 degrees C and also at 0 degrees C, while in the absence of Mg2+ the amounts of 14C and 32P incorporated were similar for both substances. Autoradiography of the SDS-polyacrylamide gel after electrophoresis of the 32P-ATP-treated membrane protein showed two distinct zones corresponding to protein bands. Sonication released twice as much 32P-ATP as 14C-ATP from the space within the membrane particles indicating that at least half of the ATP present in space did not contain its original terminal phosphate group. About 40--45% of the 32P-radioactivity was incorporated in the membrane lipids, whereas only small amounts of 14C-radioactivity were extracted with lipids. About 1/3 of the incorporated 14C-radioactivity was not extractable with acids. The same amount remained in the 32P-ATP treated preparation acid-stably bound after extraction of the lipids and hus must be firmly bound ATP. When the reaction of the membrane preparation with labelled ATP was performed at 0 degrees C the fractions of the acid-stably bound 32P- and 14C-radioactivity increased. About 1 nmole/mg of protein (10--15%) of the bound 32P-radioactivity was exchangeable against unlabelled ATP, while only a very small fraction (less than 0.5 nmol/mg protein) of the 14C-radioactivity was exchanged against unlabelled ATP. Preincubation of the membrane particles with ATP-Mg2+ at 0 degrees C induced 30% inhibition of the ATPase activity and abolition of the net uptake of catecholamines. Different Km values obtained from initial velocity studies of ATPase activity and the overall-incorporation of 32P-radioactivity indicated that a direct correlation between these processes did not exist. Different strong inhibitory effects exerted by ADP on the ATPase activity and net uptake of catecholamine at the one hand and the overall 32P-and 14C-incorporation at the other hand supported that view. It is concluded that small fractions of the observed 32P-and 14C-incorporation can be involved in the ATP hydrolyzing reaction.


Histamine receptors in the guinea pig ileum. Histamine and some related compounds acting selectively on H2-or H1 receptors were tested for their ability to contract the guinea pig ileum, in the usual whole ileum preparation and in the longitudinal muscle preparation. The concentrations elicited by histamine in both kinds of preparations were not potentiated by cimetidine or metiamide and were not inhibited by administration of H2 receptor selective agonists in doses which were subthreshold for contracting the guinea pig ileum; higher doses of the H2 agonists could actually potentiate the effect of histamine. The results obtained suggest that H2 receptors with relaxing effect do not occur in the guinea pig ileum or at least that they are not involved in the contraction of the longitudinal muscle layers. The possibility that a sub-type of H2 receptors with properties different from those of the "classical" H2 receptors so far known, exists in the guinea pig ileum, cannot be excluded.


Effect of fenfluramine administration on synaptosomal uptake of some neurotransmitters and on synaptosomal enzymes which metabolise GABA. Fenfluramine has been considered to deplete neuronal 5-hydroxytryptamine (5-HT). This compound is able to reduce the synaptosomal uptake of 5-HT and other neurotransmitters such as GABA and glutamic acid (Glu). The effects of fenfluramine on these three compounds considered as neurotransmitters are different. The inhibition is of competitive type for 5-HT and non-competitive for GABA and Glu. Concerning the enzymes involved in GABA synthesis and degradation, Fenfluramine increases Glutamic acid decarboxylase activity and decreases GABA-trasaminase activity in synaptosomes. Decreased synaptosomal GABA levels could be attributed to a lower uptake. An enzymatic regulating system may be responsible in restoring the GABA level. A similar mechanism concerning serotonin has been previously suggested (Costa et al., 1971).


Digoxin induced release of creatine kinase from isolated guinea-pig hearts. In isolated perfused guinea-pig hearts, digoxin produced a concentration dependent release of creatine kinase (ATP-creatine-transphosphorylase; CK). A corresponding decrease of the CK activity in the myocardium was obtained. The enzyme release seems to be a sign of glycoside intoxication, as its extent paralleled the severity of digoxin induced arrhythmias. Especially high CK activities were liberated when ventricular fibrillation occurred. Likewise, electrically induced fibrillation, in control hearts, led to enzyme release. However, the digoxin effect was not matched. Reserpine pretreatment antagonized the CK release by electrical fibrillation, whereas it increased excessively the enzyme liberating effect of higher digoxin concentrations. Also, propranolol decreased the enzyme release due to electrical fibrillation. The glycoside induced CK liberation, however, was not diminished, although the ventricular fibrillation was prevented. Increase of the potassium concentration of the perfusion fluid prevented the glycoside induced fibrillation, and reduced the enzyme release. The significance of the enzyme loss from the myocardium, and the mechanisms of enzyme release are discussed.


Iodine labelling of sea anemone toxin II, and binding to normal and denervated diaphragm. 1. Sea anemone toxin II (ATX II) which keeps the activated sodium channels open, can be labelled at its histidine residues with 125I up to a specific radioactivity of 500 Ci/mmole. Upon intraventricular injection in mice, ATX II causes acute, short-lasting hyperexcitation and convulsions. Its LD50 in mice is between 25 and 50 ng of the native peptide, and between 50 and 100 ng of the radioactive material per animal. 2. The labelled peptide is bound to mouse diaphragm from where it can be displaced by ATX II and, even better, by scorpion neurotoxin but not by other basic peptides, e.g., histone or aprotinin. Binding is not significantly influenced by 50 mM potassium, by replacing sodium with choline, by veratridine or tetrodotoxin. In contrast to binding of alpha-bungarotoxin, binding of ATX II is not changed by denervation of the diaphragm. ATX II binds not only to the muscular but also to the tendinous moiety of the mouse diaphragm. 3. ATX II lowers the surface tension of water. Further experiments are needed to establish the usefulness of 125I-ATX for labelling sodium channels in excitable membranes.


Metabolic turnover of pyridine nucleotides in ascites cells of sarcoma Sa 180 and in the liver tissue of rats before and after ionizing radiation. Metabolic turnover of NADP+ labeled with 14C in the ribose moiety of their molecules was determined in the ascites cells of sarcoma Sa 180 and in the rat liver tissue. The half-lives of NAD+ and NADP+ in the Sa 180 sarcoma cells were 60 min (NAD+) and 90 min (NADP+) and 120 min (NADP+). Experiments were done on animals to study the time-dependent effect of ionizing radiation on the values of pooled NAD+ and NADP+ after 0.129 C/kg (500 R) and 0.387 C/kg (1500 R) whole-body irradiation, and the metabolic turnover of these nucleotides 5 h after whole-body irradiation with 0.387 C/kg (1500 R). Exposure to 0.129 C/kg (500 R) whole-body irradiation induced no apparent changes compared to the controls. Within 5 h of irradiation the whole-body dose of 0.387 C/kg (1500 R) produced changes in rat liver cells characterized by a reduction of the nucleotide biological half-lives (NAD+ from 80 to 60 min and NADP+ from 120 to 70 min). No such changes in the pyridine nucleotide turnover were detected in Sa 180 ascites sarcoma cells.


Crop-sac response after systemic and intraventricular administration of neuroleptic drugs. Present experiments were aimed at studying in pigeons the effects of some neuroleptic agents given systemically or into the 3rd cerebral ventricle on PRL secretion and following morphological changes of the crop-sac mucosa both by classical histological methods and scanning electron microscopy. In addition, such changes were also evaluated on the basis of a semiquantitative method using a 1-4 rating scale. 3- or 5-day systemic treatment with reserpine, haloperidol and (ł/-)-sulpiride produced an intense crop-sac response consisting of a marked epithelial hyperplasia and presence of milk-like material. Similarly, a much lower dose of haloperidol, clozapine, and the two enantiomers of sulpiride given into the 3rd cerebral ventricle for 3 consecutive days produced a marked crop-sac response. The l-sulpiride was more active in comparison to the d-enantiomer. In conclusion, present experiments show that, similarly to mammals, in pigeons neuroleptic drugs are able to stimulate prolactin secretion and suggest that these effects are mediated through an action at the hypothalamic and/or pituitary level by removing a tonic dopaminergic inhibition.


Metastasis of a rectal carcinoid to the posterior fossa. We report the metastasis of a rectal carcinoid to the dura mater of the posterior fossa. The clinical presentation was unusual because of a 10-year latent period between resection of the primary rectal tumor and symptoms and signs of the posterior fossa metastasis; the radiological findings were atypical of a metastasis. Gastrointestinal carcinoids, apudomas, and the concept of the APUD endocrine cell family are reviewed.


[Pindolol in the treatment of arrhythmias caused by exertion]. In this study the anti-arrhythmic effectiveness of pindolol has been tested on ventricular effort arrhythmias in a group of 22 cardiopathic patients. The drug, administered for a week at the dose of 5 mg three times a day, obtained positive results due to its beta-blocking action and its intrinsic sympathomimetic activity.


[Functional respiratory and blood gas analytical studies of the effects of fenspiride, in oral and intramuscular administration, in chronic bronchopneumopathic subjects]. A group of 20 chronic bronchopneumopathics was treated for 15 days with fenspiride orally and i.m. The behaviour of a set of functional respiratory and haemogasanalytic parameters was monitored at various times (basic, 5th, 10th and 15th days). Progressive, significant improvements in VC, FEV1, RV and in related parameters were observed. These were attributed to the drug's anti-inflammatory effect in the respiratory ways as well as to its direct antibronchospastic action. Stress is laid on the excellent clinical tolerance of fenspiride following its oral and i.m. administration.


[Bone marrow transplantation in severe aplastic anemia and acute leukemia]. Much progress has been made in allogeneic bone marrow transplantation for severe aplastic anemia (SAA) and acute leukemia (AL). In SAA it was shown that hemopoietic chimerism and apparently permanent cures can be achieved in the majority of patients by conditioning with cyclophosphamide followed by bone marrow transplantation (BMT) from an HLA-identical sibling. The previous transfusion history is crucial for failure or success: untransfused patients do very well while graft rejection is an enormous problem in most polytransfused ones. We have shown that most patients without HLA-identical sibling donors can be adequately helped as well. After conditioning with ALG followed by transfusion of haploidentical marrow and low dose androgens there is partial to complete autologous hemopoietic reconstitution in virtually all patients. This points to the fact that most of these patients have pluripotent hemopoietic stem cells that are intact, but apparently unable to differentiate to mature cells, because they are inhibited by autoimmune mechanisms. The results of BMT in patients with endstage leukemia are modest. New pilotstudies with early marrow grafts, i.e. for ANLL in first remission and for ALL in second remission indicate that with this type of approach potentially over 50% of all patients with HLA-identical siblings can be cured. We recommend that HLA-typing should be performed early in families with SAA and AL and that the possibility of a marrow graft should be seriously considered before the patients have endstage disease. Marrow grafts are technically simple but they may pose enormous problems such as graft versus host reaction (GvH), interstitial pneumonia, graft rejection and leukemic recurrence. Therefore, the procedure should only be performed in highly specialized centers with much knowledge and experience in the immunobiology of bone marrow transplantation.


[Prophylactic alkalization of the urine during cytostatic tumor treatment with the oxazaphosphorine derivatives, cyclophosphamide and ifosfamide]. The so-called cystitis due to cyclophosphamide (Cytoxan) is caused by direct contact of the mucosa with alkylating metabolites in acid urine. These alkylating metabolites can be inactivated by instillation of cysteine into the urinary bladder. The cytostatically active metabolites of ifosfamide (Holoxan), a derivative of oxazaphosphorine, are eliminated by the kidneys as well. Their special toxicity is much higher than the toxicity of Cytoxan. The alkylating metabolites of ifosfamide cause urological complications essentially in supravesical areas (tubulopyelo-ureteritis). Some clinical trials demonstrate that increase of diuresis and alkalinization of urine by orally administered Uralyt-U are able to decrease concentration and aggressiveness of those metabolites.


Conservative treatment of chronic renal insufficiency in children. Conservative management of chronic renal failure in children is essentially based on dietary prescription including recommendations for high caloric intake and a certain limitation of protein intake according to GFR in order to avoid any extra loading with nitrogen wastes. Prescriptions for sodium potassium and water have to be adjusted on their residual output. Prevention of osteodystrophy needs supplement of calcium, chelation of phosphorus with aluminium hydroxide and the prescription of vitamin D or its active derivatives. High blood pressure when present must be carefully controlled. Drugs, when necessary, have to be given with a dosage taking into account the level of renal failure. Finally, the mode of life of the uremic child should be as close to normal as possible.


An experimental model of hyper-irritability in the trigeminal skin field of the rat. Cutaneous hyper-irritability was produced in the trigeminal sensory distribution of chronically prepared rats by injecting various central stimulants into the dorsal subarachnoid space overlying caudal medulla. Effective substances were classified into three groups according to the behavioral patterns induced. These were (1) picrotoxin and penicillin G-K, (2) strychnine and brucine, and (3) D,L-homocysteic acid. Of these substances, picrotoxin was the most effective. The hyper-irritability elicited by picrotoxin or penicillin G-K was associated with a relatively localized trigger zone in the face, while that elicited by strychnine or brucine spread beyond the trigeminal region to the first cervical dermatome. D,L-homocysteic acid evoked only spontaneous scratching and vocalization. Picrotoxin-induced hyper-irritability involved facilitation of impulse transmission from cutaneous afferent terminals to second-order neurons in the trigeminal nucleus caudalis.


Endogenous opioids and fever: a hypothesis. The explosion of knowledge set off by the characterization of endorphins and their receptors has created opportunities for new perspectives on a variety of phenomena. This brief essay examines one such phenomenon--the human febrile response--and proposes an alteration of the current model of the neuroendocrine circuitry controlling body temperature. Using behavioral thermoregulation as a starting point, we shall touch upon implications of this model for describing other forms of goal-directed motor activity.


Effect of an increase in HbO2 affinity on the calculated capillary recruitment of an isolated rat heart. The effect of an increase in hemoglobin O2 affinity on myocardial O2 delivery was studied in a blood perfused working rat heart preparation. In a first series of experiments P50 (PO2 for which saturation is 50%) was lowered by use of carbon monoxide. The heart was alternatively perfused with the blood sample of P50 = 32 mm Hg and the blood sample of P50 = 17 mm Hg. O2 capacity of both samples was kept the same by appropriate hemodilution. In a second serie of experiments change of P50 was obtained by the use of adult human erythrocytes containing hemoglobin creteil with a P50 of 13.6 mm Hg. As P50 decreased from 25 to 10 mm Hg, coronary sinus PO2 (PcsO2) diminished from 26 +/- 2 to 18 +/- 2 mm Hg (-29 +/- 2%), coronary sinus O2 content (CcsO2) increased by 15 +/- 3%, myocardial oxygen consumption did not change significantly. The percentage of increase of coronary flow was 23 +/- 4%. Analysis of these results with a simple mathematical model of O2 delivery suggest that increase in HbO2 affinity is corrected by a simultaneous increase in coronary flow and capillary recruitment.


Perivascular pH and pial arterial diameter during bicuculline induced seizures in cats. The aim of the present study was to correlate locally at the same pial artery the vascular reaction with the perivascular pH during the initial phase of functional hyperemia. As a model of functional hyperemia, bicuculline (3 mg/kg i.v.) induced seizure was taken. Normally, a strong increase of blood pressure occurs together with the start of seizure. Since a discrimination between metabolically induced and pressure dependent vascular reactions is not possible under such conditions, the cats (anesthetized with 40--50 mg/kg chloralose) received in addition 3 mg/kg phentolamine and 10 mg/kg pentobarbital. Under these conditions a significant increase of blood pressure started only 50 s after the onset of seizure. Perivascular pH was recorded using spear type pH microelectrodes in the subarachnoid space surrounding a pial artery. The diameter of the respective artery was measured continuously. After onset of seizure an immediate, increasing perivascular acidosis developed which was accompanied by an increase in pial arterial diameter. The maximal decrease of pH was 0.29 units and occurred 30 s after the start of seizure. These data show that a decrease in perivascular pH can be one factor mediating functional hyperemia in the brain.


[Tumours of the undescended testis (author's transl)]. It has now been clearly demonstrated that cryptorchidism is accompanied by a very marked increase in malignancy of the ectopic testis as well as in that in place. Amongst 80 patients undergoing surgery for carcinoma of the testis 14 (17.5%) had a past history of unilateral undescended. The risk of malignancy developing in an ectopic testis is 12 to 48 times greater than that of a testis which has descended normally. It would appear that the higher the position of the testis, the greater is the risk. Orchidopexy may decreased the risk if performed before the age of 11. When the tumour is situated in an ectopic testis, the clinical picture is often misleading. When the ectopic testis has been brought down the special feature of these tumours is the frequency of spread to inguinal lymph nodes. In terms of its basic principles the treatment of these tumours does not differ from that of those affecting normal testis. The aetiology of these tumours remains uncertain. There would seem to be a consensus in favour of a disgenetic origin, possible secondary to hormonal deficiency.


[The bacteriological diagnosis of pneumonia by transtracheal puncture. Value in medical intensive care (author's transl)]. Seventy five patients referred with a diagnosis of pneumonia underwent transtracheal puncture. In 76% of cases this examination led to discovery of an organism in infected patients. In 50% of cases, the bacteriological diagnosis was sufficiently accurately oriented by direct examination to permit rapid and effective treatment. There was virtually perfect agreement with the results of blood cultures when the latter were positive. The organisms most often responsible were Gram positive and above all the pneumococcus. These results led to narrow spectrum antibiotic treatment appropriate for the organism. This early treatment, based upon the results of tracheal puncture, was associated with a favourable course in 85% of cases, in particular in severely ill patients requiring intubation. No notable complications occurred.


Alkaline stability of guanosine and some of its derivatives modified by the carcinogen N-acetoxyacetylaminofluorene. The alkaline treatment of Guo, dGuo, dGMP and denatured DNA modified by N-acetoxyacetylaminofluorene (N-AcO-AAF) was performed in 0.1 M NaOH at 40 degrees C. The kinetics of the reaction were followed by ultraviolet absorption and by chromatographic methods and were found different for the four products under study. Circular dichroism spectra show differences in the environment of acetylaminofluorene residue in these products. The alkaline treatment of Guo-AAF (and dGuo-AAF) leads to the formation of three products. These products were separated by thin layer chromatography and by HPLC and were characterized by spectroscopic methods. One is the already known unstable Guo-AF (and respectively dGuo-AF) (1). The other two products are relatively stable products of the transformation of Guo-AF (or dGuo-AF). These last ones present almost identical ultraviolet absorption spectra, but very different circular dichroism spectra.


Effect of histamine H2-receptor antagonists on concentration of noradrenaline, dopamine, 3,4-dihydroxyphenylacetic acid, serotonin and 5-hydroxyindoleacetic acid in rat brain. Intraventricular (ivt) administration of 50 micrograms of burimamide (BUR) or 250 micrograms of either metiamide (MET) or cimetidine (CIM) decreased noradrenaline (NA) concentration in the rat hypothalamus by nearly 30%. CIM did not significantly influence either dopamine (DA) or 3,4-dihydroxyphenylacetic acid (DOPAC) levels in the striatum. Serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) contents in the hypothalamus were not significantly affected by either MET or CIM.


Two noradrenergic systems in the brain and their interactions with other monoaminergic neurons. Numerous anatomical, neurophysiological and pharmacological data indicate on the existence of two functionally different noradrenergic systems in the brain: the dorsal bundle and the ventral bundle. It is possible that dorsal bundle inhibits serotonergic neurons whilst tonically stimulates dopaminergic neurons. On the other hand, the ventral noradrenergic bundle seems to exert opposite influences. New theory of affective disorders is proposed.


[Utilization of maltose during shorttime parenteral nutrition (author's transl)]. Twelve metabolically healthy patients, who had to undergo cholecystectomy or gastric resection, were fed intermittently by the parenteral route (via veins of the upper limbs) post-operatively up to and including the 2nd post-operative day with a combined maltose-glucose solution in parallel with an 8% mixture of crystalline L-aminoacids. At an administration rate of 150 g maltose/day and 250 g glucose/day with the simultaneous administration of 80 g aminoacids, the mean carbohydrate utilisation was 82,5%. The glucose and maltose levels in the blood fluctuated in accordance with the infusion periods and the free fatty acids levels measured were their mirror image. On the whole free fatty acids were lowered significantly, which indicates adequate carbohydrate supply with consequent inhibition of lipolysis. The degree of tolerance was determined objectively by measurement of the acid-base parameter, gamma-GT, alkaline phosphatase, GOT, bilirubin, uric acid, creatinine and serum electrolytes. No changes were found which could be attributed to the administration of the infusion solution.


[Influences of different anaesthetic techniques on serum levels of hepatic enzymes and of bilirubin (author's transl)]. The influence of four different types of anaesthesia -- halothane and enflurane inhalation anaestheis, neurolept analgesia and epidural analgesia with single shot technique -- on the serum levels of GOT, GPT, GLDH, LDH, AP, LAP, gamma-GT, total protein and bilirubin was examined in 104 comparable cases undergoing gynaecological surgery. A significant increase in GOT, GPT, GLDH, AP, and gamma-GT levels was observed between the 6th and 9th postoperative day with all types of anaesthesia. The serum levels remained elevated until the 15th postoperative day. There was no significant difference between the four types of anaesthesia as regarded their effects on the serum constituents investigated. The same applied to the serum CHE activity which reached a minimum on the 3rd postoperative day followed by a steady rise. The results indicate that the postoperative changes of the liver enzyme pattern are not related to the type of anaesthesia used. Bilirubin was elevated on the first postoperative day, then dropped rapidly and stayed below the pre-operative values until the 12th postoperative day. There was no difference between the four types of anaesthesia. Glucuronisation of bilirubin may be inhibited initially by the anaesthetic agents. The subsequent rapid fall of the bilirubin levels may be the result of enzyme induction.


[The effects of "atraumatic" surgery in halothane anaesthesia and neurolept analgesia on the liver enzyme pattern (author's transl)]. The influence of general anaesthesia for operations devoid of substantial tissue traumas on the postoperative pattern of hepatic enzymes was studied in 40 patients undergoing ophthalmologic surgery. 20 had neurolept analgesia, 20 had halothane anaesthesia. The duration of anaesthesia, age of the patients and their previous history corresponded fairly closely to those of a group of patients who had gynaecological operations and were the subject of a previous study. In contrast to the latter group total protein, cholinesterase, GOT, GPT, LDH, GLDH, AP, LAP and gamma-GT remained normal up to 12 days after the operation. The exception were changes in the total bilirubin levels which were similar to those observed in the gynaecological cases. Possible causes are discussed.