Oxygen uptake and lung function in mice infected with Streptococcus pneumoniae, influenza virus, or Mycoplasma pulmonis. Model systems of respiratory infection in mice were established with Streptococcus pneumoniae, influenza virus, and Mycoplasma pulmonis. The LT50 for S. pneumoniae was 2 1/2 days, for lethal influenza 6 days, and for M. pulmonis 5 days. Morbidity in sublethal influenza infections reached a peak during days 5 to 10, with recovery indicated by the third week. The course of each pulmonary infection was followed by use of the animal's maximal ability to consume oxygen (VO2max by determining the weight, compliance, and stability of the excised lung, and in some cases by following O2 consumption of minced tissue. Depression of VO2max began early in each infection; reductions ranged from 9% at the peak of sublethal influenza infection to 50% 12 to 48 hr before the LT50 of fatal infections. The depressions were not relieved by 100% O2. The noninvasive VO2max test, evoked by cold air, was simple, rapid, and reproducible and appeared to serve as a quantitative measure of over-all function during infection. Each type of infection caused an increase in lung weight, with the largest noted during fatal Mycoplasma illness and lethal influenza. The effects on lungs by influenza and M. pulmonis infections were similar but could be differentiated from those with S. pneumoniae. With sublethal influenza, CL was reduced 30% between days 5 to 10, with recovery by the third week. Ctis was not affected. M. pulmonis infections and lethal influenza caused depressions in CL of over 60% by day 4 but only a 30% decrease in Ctis. The data suggest that the decreased compliance in influenza and M. pulmonis infections was due primarily to increased surface tension. In contrast, S. pneumoniae did not affect compliance.


Glucose 6-phosphate dehydrogenase variants: Gd (+) Alexandra associated with neonatal jaundice and Gd (-) Camperdown in a young man with lamellar cataracts. Two male subjects are described, with unusual clinical presentations and with hitherto undescribed G6PD variants. The first, of Italian extraction, suffered from severe neonatal jaundice following maternal ingestion of fresh broad beans (Vicia fava) both prenatally and postnatally: the expression of the enzymatic defect was much more severe in the neonatal period than on retesting in adolescence, when biochemical characterization showed unique features which justify designation as a new variant Gd(+) Alexandra. The second patient, a boy of Maltese extraction who was found to have bilateral lamellar cataracts at the age of 4 years, was identified as G6PD deficient only as a result of a survey of children of Mediterranean origin with unexplained cataract formation; he has approximately 15% of normal enzyme activity, with another unique combination of biochemical characteristics which has led to its designation as Gd(-) Camperdown. Although this association may be coincidental, it prompts further attention to the possibility that under certain circumstances G6PD deficiency may favor cataract formation. The two cases illustrate the value of characterization of the mutant enzyme whenever unexpected clinical or laboratory results are obtained.


Isolation and characterization of a phospholipase A2 from an inflammatory exudate. Sterile peritoneal exudates produced in rabbits injected with 1% glycogen contain a phospholipase A activity in a cell-free supernatant fraction that hydrolyzed a synthetic phospholipid (1,2-diacyl-sn-glycero-3-phospho-ethanolamine) and phospholipids of autoclaved Escherichia coli. This phospholipase activity (phosphatidylacylhydrolase EC 3.1.1.4) exhibited an apparent bimodal pH optimum (pH 6.0 and pH 7.5) and was Ca(2+)-dependent; Mg(2+) and monovalent cations (Na(+) and K(+)) did not substitute for Ca(2+) in the reaction; EDTA was a potent inhibitor. The phospholipase hydrolyzed 1-[1-(14)C]palmitoyl-2-acyl-sn-glycero-3-phosphoethanolamine to form only radio-active lysophosphatidylethanolamine as the product, indicating that the enzyme had phospholipase A(2) specificity. The phospholipase A(2) was purified 302-fold by two successive chromatographic steps on carboxymethyl Sephadex. Gel filtration (Sephadex G75) of the purified enzyme resulted in a single peak of biological activity with a molecular weight of approximately 14,800. The same estimate of molecular weight was obtained by SDS-polyacrylamide gel electrophoresis, which yielded a single band. Polyacrylamide gel electrophoresis of this fraction at pH 4.3 revealed a single protein band migrating beyond lysozyme, with the dye front, suggesting that this protein was more basic than lysozyme (pI 10.5). The enzymatic and physical-chemical characteristics of this soluble enzyme were remarkably similar to a recently described phospholipase A(2) of rabbit polymorphonuclear leukocytes derived from glycogen-induced peritoneal exudates. The possible origin and physiological role of this soluble enzyme are discussed.


CO2-inhibition of the amplitude of bending of triton-demembranated sea urcin sperm flagella. Demembranated sea urchin spermatozoa were reactivated in solutions containing KHCO3 and observed in a covered well slide. Although KHCO3 itself causes a small inhibition of flagellar beat frequency, the results confirm previous observations of a direct inhibition of flagellar bend angle by CO2 with no effect of CO2 on frequency. Observation of the effect of pH on the inhibition of bend angle in solutions containing KHCO3 indicates that a given concentration of OH- has a similar effect to the same concentration of HCO3-, as would be expected if CO2- inhibition results from reaction of CO2 with protein-NH3+ groups to form carbamates. CO2 may interfere with a control mechanism which selectively suppresses dynein cross-bridge activity in order to generate rhythmic bending. This control mechanism may incorporate a feedback control involving a measure of flagellar amplitude, which fails to operate successfully when the amplitude is reduced below a critical level.


Differences in the outcomes of acute episodes of care provided by various types of family practitioners. This study was designed to compare the outcomes achieved in a series of acute care episodes by different levels of family practice providers working in the clinic setting. The study utilizes a method which depends upon the provider to estimate level of function expected and earliest date of recovery for each episode. When the patients are viewed as a single group, those patients treated by the medex appear to fare considerably better and those seen by a faculty member do worse; however, when each functional status group is examined separately, only the asymptomatic but clinically ill patients (45 cases) show a statistically significant difference in outcomes among the providers, with the medex having good results and the faculty poor results.


Sodium and calcium movements in dog red blood cells. Determinants of 45Ca influx, 45Ca efflux, and 22Na efflux were examined in dog red blood cells. 45Ca influx is strongly influenced by the Na concentration on either side of the membrane, being stimulated by intracellular Na and inhibited by extracellular Na. A saturation curve is obtained when Ca influx is plotted as a function of medium Ca concentration. The maximum Ca influx is a function of pH (increasing with greater alkalinity) and cell volume (increasing with cell swelling). Quinidine strongly inhibits Ca influx. Efflux of 45Ca is stimulated by increasing concentrations of extracellular Na. 22Na efflux is stimulated by either Ca or Na in the medium, and the effects of the two ions are mutually exclusive rather than additive. Quinidine inhibits Ca-activated 22Na efflux. The results are considered in terms of a model for Ca-Na exchange, and it is concluded that the system shows many features of such a coupled ion transport system. However, the stoichiometric ratio between Ca influx and Ca-dependent Na efflux is highly variable under different experimental conditions. Because the Ca fluxes may reflect a combination of ATP-dependent, outward transport and Na-linked passive movements, the true stoichiometry of an exchanger may not be ascertainable in the absence of a specific Ca pump inhibitor. The meaning of these observations for Ca-dependent volume regulation by dog red blood cells is discussed.


The stimulating effect of fatty acids and amino acid derivatives on the labellar sugar receptor of the fleshfly. Seven D-amino acids, including D-valine, D-phenylalanine, D-leucine, D-isoleucine, D-tryptophan, D-methionine, and D-alpha-aminobutyric acid, are markedly less stimulative than the corresponding L-isomers that can stimulate the labellar sugar receptor of the fleshfly. A distinct effect of len;th of the amino acid side chain is clearly observed. Esterification and amidation of the alpha-carboxyl group, as well as substitution by hydroxyl and methyl groups, result in extremely decreased responses. Amino acids whose amino groups are located at a position other than the alpha are almost ineffective. With all these rigid stereospecificities of the sugar receptor for amino acids, certain replacement of the alpha-amino group with the hydroxyl or carbonyl group shows a slight increase of the response at neutral pH. Furthermore, certain fatty acids can stimulate the sugar receptor once the solutions are buffered at neutral pH. This observation was further supported by the presence of a remarkable similarity of stimulating effectiveness between amino acids that can stimulate the sugar receptor and those fatty acids. The similarity was shown by testing the response concentration relationships, the stimulating effect of fatty acid derivatives, the effect of treatment with p-chloromercuribenzoate, the behavioral response, and so on.


On the effects of divalent cations and ethylene glycol-bis-(beta-aminoethyl ether) N,N,N',N'-tetraacetate on action potential duration in frog heart. Resting and action potentials were recorded from superfused strips of frog ventricle. Reducing the bathing calcium concentration ([Ca2+]0) with or without ethylene glycol-bis(beta-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA) prolongs the action potential (AP). The change in the duration of the AP extends over many minutes, but is rapidly reversed by restoring calcium ions. Other changes (e.g., in resting potential and overshoot) are, however, only more slowly reversed. Reducing [Ca2+]0 with 0.2, 2, or 5 mM EGTA produces progressively greater prolongation of AP; maximum values were well in excess of 1 min. This prolongation can be reversed by other divalent cations in EGTA (Mg2+, Sr2+) or Ca-free (Mn2+) solutions, or by acetylcholine. Barium ions increase AP duration in keeping with their known effect on potassium conductance. D600, which blocks the slow inward current in cardiac muscle, is without effect on the action potentials recorded in EGTA solutions, or on the time course and extent of the recovery to normal duration upon restoring calcium ions. It is concluded that divalent cations exert an influence on membrane potassium conductance extracellularly in frog heart. The cell membrane does not become excessively "leaky" in EGTA solutions.


Characterization of L-aspartate uptake by Streptomyces hydrogenans. Multiple transport systems for L-aspartic acid exist in Steptomyces hydrogenans. The intracellular accumulation of L-aspartate against a concentration gradient was immediately inhibited by proton conductors, such as carbonyl cyanide p-trifluoromethoxyphenylhydrazone, 2,4-dinitrophenol or nigericin. Transport activity was gradually lost when inhibitors of protein synthesis were added. L-Aspartate transport had two pH optima at 6.5 and 4.5. At pH 6.5, two saturable transport components with different Km and Vmax values could be resolved by kinetic studies. A high-affinity system (system I) preferred the L-isomers of the anionic forms of aspartic and glutamic acid. At the same pH, a second, low-affinity system (system II) operated, which was presumably less specific than system I and also able to accept, at high concentrations, neutral amino acids. At pH 4.5, the Lineweaver-Burk plot revealed only a single catalytic component, with Km and Vmax values similar to those of system II. Again, in contrast to system I, this component showed high affinity for neutral amino acids. The data suggest that L-aspartic acid and L-glutamic acid are transported by this system as neutral zwitterionic molecules.


The mode of action of N-(n-Dodecyl)diethanolamine with particular reference to the effect of protonation on uptake by Escherichia coli. In a homologous series of N-(n-alkyl)diethanolamines antimicrobial activity was related to surface activity and increasing octanol-water partition coefficient. Maximum activity was exhibited by the dodecyl-, tetradecyl- and hexadecyl-derivatives. Dodecyldiethanolamine (DDE) displayed a broad spectrum of activity. Towards Escherichia coli NCIB8277, its bacteriostatic and bactericidal activity increased as the degree of protonation lessened, and may have been influenced by the formation of micelles. Uptake of DDE by washed suspensions of E. coli was more rapid and more extensive at pH 7.0 than pH 4.0. Within this pH range, bacterial uptake, the octanol-water partition coefficient (lipid solubility) and the proportion of unprotonated DDE all increased. Uptake isotherms at pH values in the range 4.0 to 8.0 are interpreted as signifying different uptake mechanisms for the protonated and unprotonated forms.


Comparative digestibility of acid detergent fiber by laboratory albino and wild Polynesian rats. The effects of adding 5%, 10%, and 15% acid detergent fiber to a nonfibrous basal diet were examined in a comparative feeding study with Polynesian rats (Rattus exulans) and laboratory rats. Digestibility coefficients for dry matter, crude protein, and gross energy declined significantly in both species as fiber content increased, but averaged significantly lower in the Polynesian than in the laboratory rat. Fiber digestibility was not significantly affected by fiber level but was by species, with the Polynesian rat having the higher digestibility coefficients.


Fetal hypertension and the development of increased pulmonary vascular smooth muscle: a possible mechanism for persistent pulmonary hypertension of the newborn infant. Chronic pulmonary arterial hypertension was produced in six fetal lambs. In four (126 to 139 days' gestation) unilateral fetal renal artery constriction caused systemic arterial mean blood pressure elevations. In another fetus, constriction of the umbilical artery caused a systemic mean blood pressure elevation; in the sixth, partial occlusion of the ductus arteriosus caused isolated pulmonary arterial hypertension. The right lung of each fetus was perfused with fixative at the in vivo mean arterial pressure and the amount of smooth muscle in the fifth generation (resistance) vessels analyzed using the medial width/external diameter ratio. There was a significant increase in the medial width/external diameter ratio in the six experimental animals as compared to that in six normal fetuses. In separate fetuses the increased ratios were due to a decreased external diameter, increased smooth muscle, or both these factors. The total number of resistance vessels was counted in the right lung of each fetus and no significant difference from normal was observed. We postulate that either fetal systemic hypertension or constriction of the ductus arteriosus causes fetal pulmonary hypertension in utero and that this produces increased smooth muscle development in pulmonary arterial resistance vessels; this may be a pathogenic mechanism for the syndrome of persistent pulmonary hypertension of the newborn infant.


Monitoring critically ill newborn infants with digital capillary blood samples: an alternative. Capillary blood samples obtained from a warmed distal phalanx of the right hand were compared with either temporal or right radial arterial blood samples for PO2, PCO2, and pH in 33 critically ill newborn infants. The blood pressure and skin temperatures of each infant and the ambient oxygen concentration were recorded at the time the blood was sampled. Sixty-eight paired PO2 analyses yielded a regression line close to the line of identity. The mean difference between digital capillary and arterial PO2 was 11.3 +/- 1.4 mm Hg (r = 0.92). The results were similar for the paired PCO2 analyses (r = 0.84) and for the paired pH analyses (r = 0.94). The correlation between arterial PO2 and digital capillary PO2 deteriorated when the systolic blood pressure of the patient was below 35 mm Hg. There was no correlation between skin temperature and capillary-arterial PO2 differences. The frequency of retrolental fibroplasia leading to blindness was not different from that in nurseries that sample umbilical arterial blood routinely.


Physicochemical properties of glycyrrhizic acid in aqueous media II: Effect on flocculation-deflocculation behavior of suspensions of sulfathiazole and graphite. The flocculation-deflocculation behavior of sulfathiazole and graphite in aqueous solutions of glycyrrhizic acid was studied by measuring the sedimentation volume and turbidity of supernates. The dispersing effect of glycyrrhizic acid on suspension of sulfathiazole showed a maximum in the pH 3-4 region, the same pH region where the zeta-potential of sulfathiazole particles showed a negative maximum. The results were explained by the variation of degrees of ionization of glycyrrhizic acid and sulfathiazole with pH. With graphite suspensions, the pH region where the dispersing effect of glycyrrhizic acid showed a maximum shifted to a higher pH compared with sulfathiazole. This result can be attributed to the fact that graphite is a nonpolar substance so the surface properties are not affected by a pH change. Hence, the adsorption of glycyrrhizic acid occurs even in a fairly high pH range.


Hydrolytic degradation of 2,6-dichlorobenzylthiopseudourea hydrochloride. Degradation of 2,6-dichlorobenzylthiopseudourea hydrochloride was followed in basic medium (pH 7.5) to isolate and characterize all possible degradation products. IR, Raman, and NMR spectroscopy, TLC, and elemental analysis were used to identify the products. Degradation of base-hydrolyzed 2,6-dichlorobenzylthiopseudourea hydrochloride produced 2,6-dichlorobenzylthiol and cyanamide and was followed by oxidation (air) to produce bis(2,6-dichlorobenzyl) disulfide, dimerization to give cyanoguanidine, and hydrolysis to yield urea. The kinetics of hydrolysis at 22.5 degrees (pH 7.0 and 7.5) and at 37 degrees (pH 7.0) revealed a pseudo-first-order reaction with respect to the substrate. Apparent first-order rate constants and energy of activation, entropy of activation, and half-life values were determined.


Improved colorimetric determination of aspirin and salicylic acid concentrations in human plasma. An improvement in a previously described method for the determination of plasma salicylic acid and aspirin levels in humans is described. The procedure was simplified by employing only one plasma sample for both salicylates. More accurate estimation of salicylates, particularly aspirin, was achieved by using two different calibration curves. Salicylic acid was estimated by reaction with an aqueous solution of the Folin-Ciocalteu phenol reagent. Absorbance of the blue-colored complex, which formed on addition of sodium hydroxide, was measured at 670 nm. The influence of alkalinity in the formation of the colored complex is discussed. The average recovery of aspirin added to plasma was 94.61%; it was 214.72% by the previous method.


An investigation of the ionic mechanism of intracellular pH regulation in mouse soleus muscle fibres. 1. Intracellular pH (pH(i)) of surface fibres of the mouse soleus muscle was measured in vitro by recessed-tip pH-sensitive micro-electrodes. pH(i) was displaced in an acid direction by removal of external (NH(4))(2)SO(4) after a short exposure, and the mechanism of recovery from this acidification was investigated.2. Removal of external K caused a very slow acidification (probably due to the decreasing Na gradient) but had no effect on the rate of pH(i) recovery following acidification. This indicates that K(+)-H(+) exchange is not involved in the pH(i) regulating system.3. Short applications of 10(-4)M ouabain had no obvious effect on pH(i) and did not alter the rate of pH(i) recovery following acidification. This suggests that there is no direct connexion between the regulation of pH(i) and the Na pump.4. Reduction of external Ca from 10 to 1 mM caused a transient fall in pH(i), but the rate of pH(i) recovery following acidification was unaffected. This suggests that Ca(2+)-H(+) exchange is not involved in the pH(i) regulating system.5. An 11% reduction in external Na caused a significant slowing of pH(i) recovery following acidification. 90% or complete removal of external Na almost stopped pH(i) recovery. This suggests that Na(+)-H(+) exchange is involved in pH(i) regulation.6. Amiloride (10(-4)M) reversibly reduced the rate of pH(i) recovery to much the same extent as removal of external Na. Its effect was not additive to that of removal of external Na.7. Internal Na ion concentration ([Na(+)](i)), measured using Na(+)-sensitive micro-electrodes, fell on application of (NH(4))(2)SO(4) and increased on its removal. The increase transiently raised [Na(+)](i) above the level recorded before (NH(4))(2)SO(4) application. This overshoot of [Na(+)](i) was almost completely inhibited by amiloride. This is consistent with the involvement of Na(+)-H(+) exchange in the pH(i) regulating system.8. Removal of external CO(2) or application of SITS (10(-4)M) caused some slowing of the rate of pH(i) recovery following acidification by removal of (NH(4))(2)SO(4). The effect of SITS was additive to that of Na-free Ringer or amiloride. These results suggest that Cl(-)-HCO(3) (-) exchange is also involved in the pH(i) regulating system and that it is a separate mechanism. Under the conditions used, Cl(-)-HCO(3) (-) exchange formed about 20% of the pH(i) regulating system.9. Decreasing the temperature from 37 to 28 degrees C not only caused an increase in pH(i), but also considerably slowed the rate of pH(i) recovery following acidification. We have calculated a Q(10) for Na(+)-H(+) exchange of 1.4 and for Cl(-)-HCO(3) (-) exchange, 6.9.10. We conclude that the pH(i) regulating system is comprised of two separate ionic exchange mechanisms. The major mechanism is Na(+)-H(+) exchange, which is probably driven by the transmembrane Na gradient. The other mechanism is Cl(-)-HCO(3) (-) exchange, which probably requires metabolic energy.


The role of bicarbonate, chloride and sodium ions in the regulation of intracellular pH in snail neurones. 1. Intracellular pH (pH(i)), Cl(-) and Na(+) levels were recorded in snail neurones using ion-sensitive micro-electrodes, and the mechanism of the pH(i) recovery from internal acidification investigated.2. Reducing the external HCO(3) (-) concentration greatly inhibited the rate of pH(i) recovery from HCl injection.3. Reducing external Cl(-) did not inhibit pH(i) recovery, but reducing internal Cl(-), by exposing the cell to sulphate Ringer, inhibited pH(i) recovery from CO(2) application.4. During pH(i) recovery from CO(2) application the internal Cl(-) concentration decreased. The measured fall in internal Cl(-) concentration averaged about 25% of the calculated increase in internal HCO(3) (-).5. Removal of external Na inhibited the pH(i) recovery from either CO(2) application or HCl injection.6. During the pH(i) recovery from acidification there was an increase in the internal Na(+) concentration ([Na(+)](i)). The increase was larger than that occurring when the Na pump was inhibited by K-free Ringer.7. The increase in [Na(+)](i) that occurred during pH(i) recovery from an injection of HCl was about half of that produced by a similar injection of NaCl.8. The inhibitory effects of Na-free Ringer and of the anion exchange inhibitor SITS on pH(i) recovery after HCl injection were not additive.9. It is concluded that the pH(i) regulating system involves tightly linked Cl(-)-HCO(3) (-) and Na(+)-H(+) exchange, with Na entry down its concentration gradient probably providing the energy to drive the movement inwards of HCO(3) (-) and the movement outward of Cl(-) and H(+) ions.


Further evidence for adrenergic transmission in the human vas deferens. 1. Isolated portions of human vas deferens responded to field stimulation of the intramural nerve fibres or to exogenously applied noradrenaline with rhythmical contractions of both longitudinal and circular muscle layers. 2. In guinea-pig and rabbit vas deferens field stimulation produced an initial rapid 'twitch' response which was not found with human vasa. 3. Responses of the human vas to field stimulation were depressed by the alpha-adrenoceptor blocking agents phentolamine and yohimbine. 4. It is concluded that the motor innervation of the human vas deferens is adrenergic and the relevance of this to the physiological operation of the tissue is discussed.


The effect of foreign cations, pH and pharmacological agents on the ionic permeability of an excitatory glutamate synapse. 1. Voltage clamp studies of the post-synaptic membrane of the insect neuromuscular junction have shown that normal amplitude glutamate currents could be recorded for a limited time when external Na was completely replaced by Ca, Li, ammonium, methylamine and guanidine. No change in the reversal potential of the glutamate current was observed when Na was replaced by these ions. It is suggested that the glutamate ionic channel has a similar permeability to Na and to these foreign cations, although the foreign ions cause a longer-term block of the permeability increase or receptor function. 2. Procaine, pentobarbitone, 2-4-6-triaminopyrimidine, high and low pH and low temperature reduced the synaptic ionic permeability but did not alter the ratio of the conductance increase of Na to K (delta g Na/delta gK). 4-Aminopyridine and TEA did not reduce the synaptic ionic permeability. 3. The 3. The properties of the synaptic ionic channels resemble the tight junction transepithelial ionic channels of the mammalian gall-bladder, but are very different from the Na and K non-synaptic channels of axons. It is suggested that Na and K normally pass through a single relatively large channel containing strong proton accepting acidic ligands which render the channel cation selective.


Study of the structural requirements for dopa potentiation and oxotremorine antagonism by L-prolyl-L-leucylglycinamide. A number of analogs of the tripeptide L-prolyly-L-leucylglycinamide (1) were synthesized and evaluated in the Dopa potentiation and oxotremorine antagonism tests. The replacement of the glycinamide residue with either the glycine methylamide, glycine, aminoacetonitrile, amino-2-propanone, semicarbazide, or beta-alaninamide residues resulted in a loss of activity in both tests. A 1:1 mixture of L-prolyl-L-leucyl-(-)-thiazolidine-2-carboxamide (8) and L-prolyl-L-leucyl-(+)-thiazolidine-2-carboxamide (9) showed marked activity in the Dopa potentiation test but was unable to antagonize the tremors induced by oxotremorine. L-Prolyl-L-leucyl-L-prolinamide (11), on the other hand, was active in the oxotremorine antagonism test but inactive in the Dopa potentiation test. The replacement of the pyrrolidine ring of 1 with either a thiazolidine or cyclopentane ring system caused a loss of activity. The cyclopentanecarboxylic acid analogue 13, however, was found to have moderate activity in the serotonin potentiation test.


The ability of smooth and rough strains of Streptococcus pneumoniae to activate human complement by the alternative pathway. Three capsulate pneumococcal strains of serotypes 1, 2 ans 3, and one non-capsulate strain of serotype 47, were found to activate human complement by the alternative pathway to a similar extent over the concentration range examined. Nevertheless, the capsulate strains, in contrast to the non-capsulate, are known to require complement attachment for phagocytosis and it is therefore postulated that the toxic by-products released cause the wave of oedema characteristic of pneumococcal lobar pneumonia.


The kinetic mechanism of action of an uncoupler of oxidative phosphorylation. The chemiosmotic hypothesis predicts that the mechanism by which weak acids uncouple oxidative phosphorylation in mitochondria is identical to the mechanism by which they transport hydrogen ions across artificial bilayer membranes. We report here the results of a kinetic study of uncoupler-mediated hydrogen ion transport across bilayer membranes. We made electrical relaxation measurements on black lipid membranes exposed to the substituted benzimidazole 5,6-dichloro-2-trifluoromethylbenzimidazole. The simplest model consistent with our experimental data allowed us to deduce values for adsorption coefficients and rate constants. Our major conclusions are that the back diffusion of the neutral species is the rate limiting step for the steady state transport of hydrogen ions, that both the neutral and charged forms of the uncoupler adsorb strongly to the interfaces, and that the reactions at the membrane-solution interfaces occur sufficiently rapidly for equilibrium to be maintained. Independent measurements of the adsorption coefficients of both the neutral and anionic forms of the weak acid and also of the permeability of the membrane to the neutral form agreed well with the values deduced from the kinetic study.


Endotoxin alters spontaneous transmitter release at the frog neuromuscular junction. The direct neurotoxic effects of E. coli endotoxin (ETX) on spontaneous transmitter release were tested at the frog sartorius muscle neuromuscular junction. Spontaneous transmitter release was monitored by intracellularly recording miniature end-plate potentials (MEPPs). Junctions were continuously exposed to standard concentrations of 10 microgram/ml of 3 ETX samples, 2 of which produced a significant elevation of MEPP frequency followed by a decline of frequency to very low rates. The third ETX sample, known to have a decreased canine lethality, was without effect on MEPP frequency. No significant changes in MEPP amplitude were evident. The rate of change in MEPP frequency, but not the peak frequency, was reduced by lowering ETX concentrations. Alterations in MEPP frequency induced by ETX were prevented by removing Ca++ and antagonized by high [K+]out. The results suggest that ETX alters ion conductance channels, particularly those for Ca++, in the presynaptic terminal membrane.


Biochemical and morphological characterization of mycobacteriophage R1. Large-scale propagation of mycobacteriophage R1 in broth culture has allowed the isolation of quantities of virus sufficient for characterization of its nucleic acid and lipid components as well as investigation of its ultrastructural attributes. Analysis of R1 DNA indicates that it is double stranded and possesses a molecular weight of 2.5 X 10(7) and a guanine-plus-cytosine content of 65.7 +/- 0.5%. The lipid fraction of R1 accounts for 14% of the total dry weight of the virus, 20% of which was identified as free or esterified sterols. A rapid loss of viral titer occurred after seconds of exposure to organic solvents. This result suggests that the lipid fractions of R1 is essential for its infectivity.  Electron microscopic investigation of solvent-extracted R1 showed extensive deterioration of its normal morphology, including nucleocapsid disintegration and base plate separation. Routine phosphotungstate preparations demonstrated a particle with an oval head and a noncontractile tail. Altering the pH of the phosphotungstate negative stain from neutrality damage the viral particles. Uranyl formate-contrasted specimens displayed an elongated hexagonal nucleocapsid with a neck region; the cross-striated tail possessed a starlike base plate.


Transfection in pneumococcus: single-strand intermediates in the formation of infective centers. Transfection has been found and characterized in pneumococcus. For replicating omega3 phage DNA extracted from infected cells, transfection was relatively efficient and rose linearly with DNA concentration and quadratically with time, according to T(T - 3.5) min(2). For mature DNA extracted from phage particles, transfection was hardly detectable below 1 mug/ml but increased about as the cube of the DNA concentration up to 100 mug/ml, and was still rising at concentrations over 200 mug/ml. The kinetics suggest a dependence on a mixed cubic function of the time of exposure of cells to mature DNA. Cell and phage DNAs competed with each other for transformation and transfection. Transfection was reduced much more strongly than transformation in cells that were deficient in the membrane-bound endonuclease required for conversion of donor duplex DNA to intracellular single strands; these data agree with the kinetic data in implying that independent entry of segments of two strands is necessary for transfection by replicating omega3 phage DNA and entry of at least three strands is necessary for transfection by mature DNA. To reconcile differing DNA concentration dependences of transfection and transformation with a common entry path, it was necessary to reexamine data on transformation and to recognize that this process continued to rise slowly through the concentration region usually described as "plateau." These results and the transfection data reflect multiple binding and nicking events that occurred on the cell surface before entry. Our conclusion is that transfection in pneumococcus occurs by association inside the cell of segments of single strands of phage DNA that have entered independently, creating gapped structures that need repair synthesis to create infective centers. Physical recombination is therefore automatically a prerequisite to transfection.


XXY Klinefelter's syndrome with bilateral cryptochidism, obesity, multiple capillary hemangiomas and telangiectasia. A case of XXY Klinefelter's syndrome associated with bilateral cryptochidism, -ultiple capillary hemangiomas and vesical telangiectasia is reported. Testicular biopsy revealed Leydig cell hyperplasia with hyalinization of the seminiferous tubular membrane. The patient presented with recurrent hematuria. Excretory urography, urethroscopy and renal angiography were normal. Cystoscopy revealed multiple telangiectasia, which was responsible for the hematuria. To our knowledge this is the first report of multiple capillary hemangiomas and vesical telangiectasia associated with calssical Klinefelter's syndrome.


Acetohydroxamic acid: clinical studies of a urease inhibitor in patients with staghorn renal calculi. The hydrolysis of urea by the bacterial enzyme urease pathologically increase urinary ammonia, bicarbonate, carconate and alkalinity. These factors contribute to the formation of urinary stones and to the virulence of bacteria. Acetohydroxamic acid, a potent inhibitor of urease, has been administered to 23 patients with staghorn renal calculi and urea-splitting urinary infection. Urinary ammonia and alkalinity has been reduced in every patient. A dose of 1.0 gm. acetohydroxamic acid daily has been well tolerated and effective for 2 to 12 months, even in patients with impaired renal function.


[Determination of the individual changes in cardiovascular system reactivity in prescribing adrenergic blockaders]. The changes occurring in hemodynamics due to the effect of alpha-adreno-blocking agent pyrroxan and beta-blocking agent obzidan were studied in 20 patients with neurocirculatory dystonia and in 20 patients with stage IIB hypertensive disease. Forty healthy persons formed the control group. The results of the investigation allow for the assumption that a pharmacological test with a record of the transthoracic rheogram and the derivatives of the central pulse tracings help in the proper choice of the drug and its dosage with the state of hemodynamics and the inotropic myocardial reaction taken into account.


Age related changes in muscle protein degradation. In vitro autolytic degradation of sarcoplasmic proteins in red, white and cardiac muscles increases with advance in age and with increase in temperature, the rate varying with age. Higher activity is seen in the alkaline range in all age groups. Ca2+ activated proteolytic activity also increases with advance in age.


Changes of adrenal catecholamines and their synthesizing enzymes during ontogenesis and aging in rats. Male Wistar specific-pathogen-free rats aged 2, 7, 17, 30, 60, 120, 200, 360 and 600 days, all killed in experiment on the same day, were examined. The body weight significantly increased until the 200th day, the weight of adrenals until the 120th day and the adrenal protein content until the 30th day of life. The adrenaline content of the adrenals increased continuously during the 600 days studied. Adrenal noradrenaline content increased rapidly over the first 17 days, remained at a stable level until the 120th day, and rose to a higher level after 200 days. The activity of adrenal catecholamine-synthesizing enzymes also increased with age: tyrosine hydroxylase gradually increased until the 360th day, dopamine-beta-hydroxylase and phenylethanolamine-N-methyl transferase until the 200th day. Our results demonstrate that, in the rat, during development there is a gradual increase of adrenal weight, adrenaline content, tyrosine hydroxylase and phenylethanolamine-N-methyl transferase activity until maturation (120th day), whereas the adrenal noradrenaline content reaches the adult values earlier, around the 17th day. During aging, adrenal catecholamines significantly increase when compared to young-adult rats (120-day-old), probably due to the elevated activity of the adrenal catecholamine-synthesizing enzymes. The increased adrenal catecholamine levels in old animals might be connected with a higher incidence of cardiovascular diseases in aged.


Meningitis: the influence of routine otolaryngologic consultation on morbidity and mortality in 290 cases. A prospective study was designed to obtain data on all meningitis patients admitted to a large county Communicable Disease Unit. The epidemiologic and clinical features of 290 cases were analyzed to ascertain the effect of routine O.R.L. consultation on the morbidity and mortality of this disease. The results of physical examination provided current statistics as to the existence of ear, nose, and throat diseases associated with meningitis.


Renal function in patients with multiple myeloma. Renal tubular and glomerular functions were evaluated in 35 consecutive patients with multiple myeloma and were correlated with changes in renal histopathology and myeloma protein patterns. All nine patients without Bence Jones proteinuria had CCr greater than 50 ml/min. In contrast 16/26 patients with Bence Jones proteinuria had CCr less than 50 ml/min and the magnitude of the Bence Jones proteinuria correlated well with the degree of renal insufficiency. Frequent abnormalities in renal tubular acidifying and concentrating ability were observed only in patients with Bence Jones proteinuria and occurred in the absence of significant reductions of glomerular filtration rate. Severely deranged renal histology was seen only in patients with Bence Jones proteinuria and consisted primarily of tubular atrophy and degeneration; glomeruli appeared normal. These data suggest that Bence Jones proteins exert a direct nephrotoxic effect at the tubular level with resultant tubular dysfunction and tubular atrophy. Glomerular filtration rate remains relatively preserved despite the significant abnormalities of tubular function. Although obstructing tubular casts were observed only in patients with severely impaired glomerular filtration rate, many patients with similarly impaired renal function had no evidence of such casts. Instead, tubular atrophy and degeneration correlated best with renal dysfunction.


X-linked recessive (Duchenne) muscular dystrophy (DMD) and purine metabolism: effects of oral allopurinol and adenylate. Data are presented which suggest that Duchenne muscular dystrophy (DMD) may have some origin in a severe deficiency of total muscle adenine nucleotides. Using double-blind techniques, this possibility was tested in 16 DMD patients by giving oral allopurinol, a synthetic inhibitor of the purine catabolic enzyme xanthine oxidase. Sublingual procaine adenylate was also briefly tested. Instances of clinical improvement quickly occurred which were statistically significant; they were accompanied by a significant increase in physical strength. These improvements have been maintained for more than 6 mo by administration of a small amount of allopurinol daily. Procaine adenylate had little effect. These results support the above view of DMD and seem to indicate that existing purines, retained and recycled after allopurinol, can sustain such improvement, and that additional adenylate is unnecessary.


[Kinetics of constitutive synthesis of polymethylgalacturonases by Penicillium digitatum as a function of the carbon source and pH]. Constitutive biosynthesis of polymethylgalacturonases by Penicillium digitatum is most active at the postexponential growth phase. Under certain conditions, the enzymes are synthesized at the constant rate which does not depend on the conditions of nutrition of the producing culture. An increase in the concentration of H+ ions at pH 3.0 changes the character of the fungal growth, prolongs the lag phase, decreases the rate of growth and increases the rate of the enzyme synthesis. The growth rate of Penicillium digitatum does not determine the rate of the enzyme synthesis. The enzyme synthesis does not depend on the conditions of the cultural growth.


A controlled comparative trial of clorazepate (Tranxene) and diazepam (Valium) for anxiety. Two groups of anxious patients were treated with either clorazepate (n = 27, 15 mg at night), or diazepam (n = 27, 5 mg three times a day), and the anxiolytic effects of the treatments compared during a 22 day period. Both drugs were effective antianxiety agents (as assessed by the Hamilton Anxiety Scale, Analogue and Rapid Symptom Check List scales). The response to drug treatment did not differ according to the type of anxiety--psychological or somatic. No significant difference was observed between the two drug treatments. Separation of patients according to their anxiety type did not change this finding. Clorazepate, when administered as a single dose at night, is an effective short term anxiolytic.


[Relationship between spore formation and synthesis of extracellular protheases in Bacillus mesentericus]. Two phases were established in the synthesis of extracellular proteases by Bacillus mesentericus 73. When the growth rate was high, the synthesis of proteases was small. The maximum rate of the enzyme synthesis was observed when the growth decelerated and the production of spores started. The synthesis of alkaline protease and the formation of spores are susceptible to nitrogen-metabolite and catabolite repression. The ability to produce spores was not found in the S variant of Bacillus mesentericus that did not synthesize extracellular proteases.


[Energy characteristics of the stages of photo-dependent transport of 14C-alanine in Halobacterium halobium R1 cells]. The energy of light is utilized by Halobacterium halobium R1 only at the very beginning of illumination at the same time as the transport of protons into the cells, as follows from comparing the data of photophosphorylation and light-dependent transport. The release of protons by the cells on further illumination suggests that the photoprocess and the accumulation of energy by the cells are being uncoupled. The primary transport of 15C-alanine in the course of illumination is caused directly by electrochemical gradient (i) whereas the secondary transport in the dark involves the energy liberated on hydrolysis of ATP which has been formed in the process of photophosphorylation (II). An indirect technique of assaying ATP from the transport of 14C-alanine into the cells makes it possible to characterize the distribution of utilized light energy between photophosphorylation (II) and transport processes (I). The bulk of energy is accumulated as ATP. The incorporation of 14C-alanine into the cells is stimulated twofold by illumination for a long time under physiological conditions in the presence of respiration. Therefore, light plays an important role in the energy balance of H. halobium R1.


[Cyclic pathway of formaldehyde oxidation in Pseudomonas oleovorans]. Operation of the dissimilatory hexulose phosphate cycle of formaldehyde oxidation in Pseudomonas oleovarans has been directly confirmed. Extracts of this typical facultative methylotroph catalyze the formation of 14CO2 from 14C-formaldehyde. The process depends on the presence of ribose-5-phosphate, NADP or NAD, and is linear in time. Oxidation of formaldehyde via formate to CO2 is of a minor importance in the energy metabolism of the methylotroph.


Increased and decreased sensitivity to carbon catabolite repression of enzymes of acetate metabolism in mutants of Aspergillus nidulans. The creA204, creB15 and creC27 mutations have been shown to cause carbon catabolite derepression of acetly CoA synthase and isocitrate lyase in Aspergillus nidulans. A recessive mutation, cre-34, which is linked to the creC gene, results in these enzymes being more sensitive than cre or wildtype strains to catabolite repression. The acetamidase levels of strains containing cre mutations have been investigated and provide support for the hypothesis that an acetate metabolite, rather than acetamide, induces this enzyme.


[Clobazam (Frisium) in everyday practice. An open six-months' study of its efficacy and tolerance (author's transl)]. Fifty patients suffering principally from neurotic and psychosomatic disturbances were treated for 6 months with an average of 30 mg clobazam (Frisium) per day. For documentation of the efficacy and tolerance a set of assessment scales and procedures from the ECDEU system (Early Clinical Drug Evaluation Unit) was compiled. The results showed unanimously the anxiolytic efficacy and good tolerance of clobaza. After the second week of treatment there was a significant reduction in the anxiety symptoms in the judgement of the doctor and patient (p less than 0.01). Parallel to the reduction of anxiety symptoms the general condition also improved. The good tolerance of clobazam was shown in the overall clinical assessment and in the recording of specific side effects, which could never be called severe in any case. Moreover, there were no suggestions of changes in tolerance of of dependency.


Efficiency and cost of primary care by nurses and physician assistants. We conducted a prospective study in a prepaid primary-care practice (health-maintenance organization) of a system in which nurses and physician assistants used protocols, and compared the efficiency and costs of this "new-health-practitioner" protocol system to a physician-only nonprotocol system. In five months, we studied 472 patients with any of four common acute complaints--respiratory infections, urinary and vaginal infections, headache, and abdominal pain; a subset of 203 patients was randomly allocated between the two systems. In the new-health-practitioner system physician time per patient was reduced by 92 per cent, from 11.8 to 0.9 minutes, and average visit costs--including practitioner time and charges for laboratory tests and medications--were 20 per cent less (P = 0.01). We conclude that this protocol system saves physician time and reduces costs.


Analysis of the beta-receptor mediated effect on fast-contracting skeletal muscle in vitro. Subtetanic contractions of the isolated extensor digitorum longus (EDL) of the guinea-pig, a fast-contracting muscle, were evoked by transmural field stimulation. Isoprenaline, adrenaline, terbutaline, and noradrenaline each caused a dose-dependent increase in the force of contraction, their potencies decreasing in that order. Tyramine was without effect in this respect. Curare depressed the contractions of EDL by about 20% but did not appreciably change the response to the beta-adrenoceptor agonists. The effects of isoprenaline and noradrenaline were blocked by propranolol (unselective) and H 35/25 (1-(p-tolyl)-2-isopropylamino-1-propanol, beta2-selective) but not by practolol (beta1-selective). Moreover, the increase in the force of subtetanic contractions of EDL produced by noradrenaline was unaffected by phentolamine. It is concluded that the adrenoceptor mediating the increase in the force of contraction of the isolated EDL is of the beta2-type and that the site of action is direct on the muscle. Its similarity to the receptor mediating the inverse effect on the slow-contracting soleus-muscle is pointed out.


Effect of 5-hydroxytryptophan on prolactin and growth hormone release in the infant rat: evidence for different neurotransmitter mediation. In 10 day-old female and male rats, administration of 5-hydroxytryptophan (5-HTP) induced a prompt elevation in plasma prolactin (Prl) and growth hormone (GH) levels. Pretreatment with 2 serotonin (5-HT) receptor blockers, methysergide (Meth) and metergoline (MCE), markedly reduced the 5-HTP-induced Prl rise but failed to alter the GH response to 5-HTP. Administration of 2 selective inhibitors of presynaptic 5-HT reuptake, 3-(p-trifluoromethylphenoxy)-N-methyl-3-phenyl-propylamine and chlorimipramine (CIM), potentiated the 5-HTP-stimulated Prl rise but significantly reduced the 5-HTP-induced GH release. Blockade of dopaminergic or alpha-adrenergic receptors by pretreatment with pimozide (Pim) or phentolamine (Phent), respectively, or central sympathectomy by intraventricularly (i.vt.) injected 6-hydroxydopamine (6-OHDA) were capable of reducing the 5-HTP-induced GH release without affecting the 5-HTP-induced Prl rise. These data indicate that in the infant rat the 5-HTP-induced Prl release is mediated via the brain 5-HT system and that a nonspecific activation of the catecholaminergic system is responsible for the GH response to the drug.


[New anti-migraine drugs used in the treatment of children and adolescents]. In recent years a number of new preparations from the group of antiserotonin agents (including cyproheptadine and carbazochrome) as well as beta-adrenergic blocking agents (propranolol) and clonidine have been introduced for prevention of migraine and similar headaches. The efficiency of these drugs was studied in a group of 9 selected cases--adolescents with severe migraine refractory to other methods of treatment. In 7 cases the improvement was noted. The results were compared with own observations on larger groups of adults and data from literature. Too small number of observed patients made impossible definite conclusions. It seems, however that in selected cases the use of these drugs in headaches of adolescents is useful.


[The human factor as a cause of flight accidents]. Four causes of flying accidents due to human factors are discussed: physical and psycological unfitness, insufficient physical and psychological training, and flight fatigue. Ways in which these factors can be eliminated or their effects diminished are indicated, including the establishment of a national aeronautical health service manned by physicians specialising in aerospace medicine.


Enzymic detection of metachromatic leukodystrophy patients and heterozygotes. Two unrelated families with metachromatic leukodystrophy have been examined for the leukocyte enzyme arylsufatase A. The enzyme activities clearly reflect an autosomal recessive mode of inherence. All four parents showed heterozygote enzyme levels 40-60 percent of the control range while the two affected children had less than 20 percent normal activity. The two sibs of one affected child were shown to be heterozygote carriers. A simple screening method for sulfatase activity in tears has been developed which distinguished between metachromatic leukodystrophy patients and a control population which included other neurological disorders. Enzyme screening on tears may also be used to detect other lysosomal storage diseases including Tay-Sachs and Fabry disease.


The treatment of peptic ulcer.

The treatment of peptic ulcer. Recent evaluation of the time honoured use of antacids and anticholinergics in the treatment of peptic ulcer disease has not revealed persuasive evidence of efficacy above that of placebo. New drugs becoming available as medical treatments of peptic ulcer disease, viz H2-receptor antagonists and E group prostaglandin analogues, show considerable promise but require more time for adequate evaluation. The surgical treatment of peptic ulcer disease has similarly evolved with the development of effective operations with minimal post surgical morbidity, but again await the test of time for adequate evaluation.


Localization of substance P-like immunoreactivity in nerves in the tooth pulp. The occurrence of substance P (SP)-like immunoreactivity was studied in dental pulps of the cat. In untreated animals SP-positive fibres were found in all areas of the pulp. Most fibres were seen in central parts of the pulp but they were also observed in relation to the odontoblasts. Single, possibly unmyelinated, or fine caliber fibres or small bundles of them were seen running close to large non-fluorescent myelinated nerves, to blood vessels or without any obvious association with either of these structures. Fourteen days after transection of the inferior alveolar nerve no SP-positive fibres were observed in pulps on the denervated side. Transection of the cervical sympathetic ganglion did not change the occurrence of SP-positive fibres. The results indicate the existence of at least two types of afferent fibres in the dental pulp of the cat. Since the tooth pulp has been demonstrated to give rise only to pain sensation when stimulated, the results give morphological support for a role of SP neurones in pain transmission.


Gamete development in malaria parasites: bicarbonate-dependent stimulation by pH in vitro. Gametogenesis in Plasmodium gallinaceum involves bicarbonate-dependent processes and requires a continuous supply of glucose (presumably as an energy source). Emergence and exflagellation of gametocytes, in vitro, occur independently of the CO2 tension but are rigidly correlated with the pH of the external medium. In bicarbonate-saline gametogenesis is initiated only if the pH exceeds 7.7. Our results suggest that gamete development of malaria parasites is stimulated when infected blood is exposed to air because the decrease in the CO2 tension of the blood causes its pH to rise.


[The sympathetic nervous system inhibition in the antihypertensive effect of beta-blockers (author's transl)]. The decrease of sympathetic activity by the beta-blocking drug, as demonstrated by the decreased electric activity of the splanchnic nerve and by the decreased urinary catecholamine reponse to tilt as well as by the decreased levels of plasma dopamine beta-hydroxylase exists not only in hypertension with elevated PRA but also in hypertension with normal or low PRA. In these latter cases the antihypertensive effect is better explained by the decrease in the sympathetic nervous system activity than by the decrease of PRA. This effect seems to be indirect and probably, as suggested by Lewis, as a result of damping sensory input to the central nervous system from the heart, whose capacity to respond to exercice and stress is blunted by beta-adreno-receptor blockade.


Haemoglobin oxygen dissociation curve in patients on regular haemodialysis. Changes in the haemoglobin-oxygen dissociation curve (Hb-O2) and the factors which influence its position were studied before and after dialysis in 7 patients on regular haemodialysis during 20 dialyses. Seven normal subjects were used as controls. Haemoglobin showed a lower than normal affinity for O2 in uraemic patients before haemodialysis (p50in vivo = 33.09 +/- 0.92 mmHg and p507.4 = 31.51 +/- 0.73 mmHg, P less than 0.001), and this could be considered as a protection against tissue anoxia. After dialysis Hb-O2 affinity at the patient's pH (p50 in vivo = 27.97 +/- 0.57 mmHg, P less than 0.001). This probably eliminates the benefits of the predialysis balance of tissue oxygenation, producing a degree of hypoxia, and may play a role in the genesis of post-dialysis symptoms. Measures should be taken to improve oxygenation of high risk patients with latent heart failure or respiratory disturbances during and after dialysis.


The effect of lesions of raphe nuclei on the cataleptic action of neuroleptics. The effect of electrolytic lesions of dorsal raphe nucleus (DRN) and median raphe nucleus (MRN), produced with two methods, was tested on catalepsy produced by spiperone, pimozide and haloperidol. Using two methods of evaluating catalepsy (acc. to Delini-Stula and Morpurgo and to Costall et al.) it has been found that both kinds of MRN lesions increase the catalepsy produced by neuroleptics, with the exception of the action of haloperidol as assayed with the Delini-Stula and Morpurgo method, and pimozide as assayed with the method of Costall et al. A lesion of DRN either slightly antagonized or did not influence the cataleptogenic action of neuroleptics.


Ionised calcium during haemodialysis.

Ionised calcium during haemodialysis. In spite of the popularity of high calcium dialysate internationally, most centres in the United Kingdom continue to use a dialysate calcium of 1.5 to 1.6 mmol/L. In eleven patients studied throughout dialysis against such dialysate we conclude that it is sufficient to raise ionised calcium. Patients so treated have little elevation of PTH pre-dialysis and it is further suppressed during dialysis.


[Nicotinamide coenzymes at the early stages of light induction of carotenogenesis in the Neurospora crassa mycelium]. Changes in the concentration of NAD+, NADH, NADP+ and NADPH in the mycelium of the NADase free mutant of Neurospora crassa were studied during the latent stage of light induction of carotenogenesis. A 30 minute illumination by visible light brought about a stable decrease in the NADH/NAD+NADH ratio, exerting no effect on the NADPH/NADP+NADPH ratio. At the same time the NADP+/NAD+ ratio increased. These changes occurred only when illumination induced carotenoid accumulation in the N. crassa mycelium.


Peptic ulceration in kidney transplantation. Gastroduodenal ulceration occurred in 45 patients during the post-transplantation period in a series of 500 transplantations of 434 patients. The mortality rate of this complication was high, 42%. Bleeding and perforation were the main problems. These complications occurred frequently during treatment for acute rejection. Present day prophylaxis, which is based on the use of antacids, seems to be inadequate for controlling these complications. Other possibilities for reducing the incidence of gastroduodenal ulceration in transplant patients are discussed. Since increased serum gastrin concentrations are often observed in these patients, prophylactic treatment should be based on preoperative evaluation of gastric secretion and serum gastrin determinations. The new histamine (H2) blocking agents should be evaluated in these patients.


Dependence of the rates of dissolution of the Fe4S4 clusters of Chromatium vinosum high-potential iron protein and ferredoxin on cluster oxidation state. The influence of oxidation state on the pH dependence of the dissolution of the Fe(4)S(4) clusters of Chromatium vinosum ferredoxin and high-potential iron protein (HIPIP) has been studied. The first-order rate constants (k(obs)) for dissolution of both the Fe(4)S(4)(S-Cys)(4) (2-) and Fe(4)S(4)(S-Cys)(4) (3-) clusters of the ferredoxin follow the same overall kinetic equation but with differing specific rate and equilibrium constants. The dependence of rate and equilibrium constants upon oxidation state may be rationalized on the basis of the accompanying change in electrostatic affinity of a cluster toward H(+) and HO(-). A more drastic change in the pH dependence of the kinetics of dissolution of the Fe(4)S(4) cluster of the HIPIP accompanies its change in oxidation state. Whereas the values of k(obs) for dissolution of HIPIP containing the Fe(4)S(4)(S-Cys)(4) (2-) cluster are strictly second order to [H(+)] and [HO(-)], the pH dependence for dissolution of the HIPIP Fe(4)S(4)(S-Cys)(4) (1-) cluster indicates a first-order dependence upon [H(+)], a second-order dependence upon [HO(-)], and a spontaneous or water rate. These reactivity differences may be related to changes in cluster charge density. Mechanisms of dissolution involve preequilibrium protonation at acidic pH and preequilibrium ligand exchange at basic pH.


Probing of beta-adrenergic receptors by novel fluorescent beta-adrenergic blockers. The synthesis of two high-affinity fluorescent beta-adrenergic blockers is described: dl-N(1)-[2-hydroxy-3-(1-naphthyloxy)propyl]-N(2)-(9-acridyl)-1,2-propanediamine (9-aminoacridylpropanolol, 9-AAP) and dl-N-[2-hydroxy-3-(1-naphthyloxy)propyl]-N'-dansylethylenediamine (dansyl analogue of propranolol, DAPN). Both 9-AAP and DAPN inhibit competitively the l-epinephrine-dependent adenylate cyclase activity [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] in turkey erythrocyte membranes without affecting the fluoride-stimulated adenylate cyclase activity. Similarly, 9-AAP and DAPN inhibit in a competitive manner the binding of [(125)I]-iodohydroxybenzylpindolol to these beta-adrenergic receptors. The two fluorescent beta-adrenergic blockers 9-AAP and DAPN probe specifically beta-adrenergic receptors in the central nervous system as well as in other organs when injected into rats. The fluorescence pattern can be monitored by fluorescence microscopy performed on cryostat slices of these organs. The appearance of the characteristic fluorescence pattern can be blocked in a stereospecific fashion by a prior injection of l-propranolol and not by a prior injection of d-propranolol. These compounds therefore offer a powerful means to map beta-adrenergic receptors in vivo. The stereospecific displacement of 9-AAP from the beta-adrenergic receptors of turkey erythrocyte membranes by l-propranolol and by l-epinephrine can be detected in vitro using front-face fluorescence. The potential use of these compounds to probe beta-receptors in vitro and in vivo is discussed.


Evidence for control of serotonin secretion from human platelets by hydroxyl ion transport and osmotic lysis. Serotonin secretion from human platelets, stimulated either by thrombin or the calcium ionophore A23187, was found to be inhibited by anion transport blocking drugs such as 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS), pyridoxal phosphate, probenecid, and suramin. These drugs have previously been shown to inhibit ATP-evoked release of epinephrine from isolated chromaffin granules by blocking chloride uptake and subsequent osmotic lysis. However, in contrast to granule release, platelet secretion was insensitive to chloride and, instead, was dependent on OH-. Platelet release was suppressed by low pH, and inhibition by the transport blocking drugs was competitive only with respect to OH-. Serotonin release from platelets was also suppressed by increasing extracellular osmotic strength, and the relationship between suppression and external osmotic strength was quantitatively similar to that observed in the case of chromaffin granules. We conclude that platelet exocytosis could occur when serotonergic granules are closely juxtaposed to the plasma membrane, thus exposing the granule anion transport site to the more alkaline medium. Secretion of serotonin could occur as a consequence of OH- transport and osmotic lysis of the granule-plasma membrane complex, analogous to the chemiosmotic mechanism of chloride-dependent epinephrine release from isolated chromaffin granules.


Experimental evidence for a kinetic and electrochemical memory in enzyme membranes. The existence of hysteresis phenomena in artificial enzyme membranes due to the coupling of simple kinetic enzyme properties with diffusion transport processes is reported. The intramembrane pH of a urease coating on the surface of a glass pH electrode exhibits a hysteresis loop when the pH of the bulk solution varies cyclically. The steady-state potential of a urease membrane, as a function of the substrate concentration in the bulk solution, also exhibits a memory effect. The influence of the membrane's history on its overall behavior is visualized by electron microscopy. We interpret the results in terms of a coupling between the enzyme reactions and diffusion processes, without taking into account molecular effects.


Identification of transmembrane bridging proteins in the plasma membrane of cultured mouse L cells. Studies were carried out to identify transmembrane bridging proteins in the plasma membrane of mouse L-929 cells. Cells grown in suspension culture were 125I-labeled by lactoperoxidase and allowed to ingest latex particles to produce inside-out membrane phagosome preparations. Phagosomes were isolated and the inner membrane surface was labeled with N-(5'-aminopentyl)-5-dimethylamino-1-naphthalenesulfonamide (dansylcadavarine) by a transglutaminase-catalyzed reaction. The phagosome membrane proteins were solubilized and dansylcadavarine-labeled proteins were isolated by anti-dansyl immunoadsorbent affinity chromatography. Dansylcadavarine-labeled proteins were analyzed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis and autoradiography for the presence of 125I-labeled material. By this technique, two iodinated proteins with molecular weights of approximately 50,000 and 80,000 appear to be selectively retained by the anti-dansyl immunoadsorbent, suggesting that these proteins span the plasma membrane.


A pH-conditional mutant of Escherichia coli. Mutants of Escherichia coli have been isolated that are able to grow on lactose at pH 7.0 but not at pH 8.1. One of these mutants was analyzed and shown to map in the Z region of the lactose operon. beta-Galactosidase (beta-D-galactoside galactohydrolase; EC 3.2.1.23) activity in toluenized mutant cells at pH 8.0 was one-tenth that at pH 7.0. Enzyme purified to near homogeneity from the pH-conditional mutant similarly exhibited pH-conditional activity under conditions where wild-type enzyme was unaffected over a pH range of 6.0-8.0. The pH-conditional beta-galactosidase was used in vivo as a probe for intracellular pH. We show that an internal pH of approximately 7.8-8.0 is maintained through an external pH range of 5.9-7.8. The phenotype of pH-conditional mutants was defined on medium with lactose as the sole carbon source. Under such conditions the gene product itself, beta-galactosidase, is required to maintain intracellular pH, since such maintenance is clearly energy-dependent. Therefore, we were able to recover a pH-conditional mutant in a cytoplasmic gene product. We predict that with any phenotype independent of energy production, however, pH-sensitive mutants will be recovered only in surface elements.


Antagonism of histamine-activated adenylate cyclase in brain by D-lysergic acid diethylamide. D-Lysergic acid diethylamide and D-2-bromolysergic acid diethylamide are competitive antagonists of the histamine activation of adenylate cyclase [ATP pyrophosphate-lyase (cyclizing); E.C. 4.6.1.1] in broken cell preparations of the hippocampus and cortex of guinea pig brain. The adenylate cyclase is linked to the histamine H2-receptor. Both D-lysergic acid diethylamide and D-2-bromolysergic acid diethylamide show topological congruency with potent H2-antagonists. D-2-Bromolysergic acid diethylamide is 10 times more potent as an H2-antagonist than cimetidine, which has been the most potent H2-antagonist reported, and D-lysergic acid diethylamide is about equipotent to cimetidine. Blockade of H2-receptors could contribute to the behavioral effects of D-2-bromolysergic acid diethylamide and D-lysergic acid diethylamide.


Heptadecapeptide gastrin in the vagal nerve. Immunoreactive gastrin was present in vagal nerves from cats, dogs, and human beings. The abdominal portion of the vagus contained gastrin in amounts ranging from 16 to 273 pmol/g of nerve tissue (wet weight). The thoracic and cervical portion of the vagi contained only minute amounts of gastrin. Gel chromatography of extracts of human, canine, and feline abdominal vagi monitored by region-specific antisera against heptadecapeptide gastrin and triacontatriapeptide cholecystokinin revealed that the vagal gastrin immunoreactivity predominantly consisted of heptadecapeptide gastrin. In addition, the vagi contained small amounts of the NH2-terminal tridecapeptide gastrin fragment as well as of the putative biosynthetic gastrin precursors, components I and II. No cholecystokinin-like molecules were demonstrable. Immunocytochemical studies demonstrated gastrin-containing nerves in the intestinal wall. The nerves were found to be most numerous in the large and distal small intestine. These findings suggest that heptadecapeptide gastrin may represent a new vagal neurotransmitter.


Prostaglandin release by slow reacting substance from guinea pig and human lung tissue. Slow reacting substance (SRS) injected into the pulmonary artery released prostaglandins E (PGE) and F2alpha (PGF2alpha) and the 15-keto-13, 14-dihydro PG metabolites from non-sensitized and ovalbumin sensitized, isolated, perfused guinea pig lungs. PGs were also released from lungs incubated with SRS. Sensitized lungs released more PGs in both types of preparations. Indomethacin inhibited the effect of SRS. Passively sensitized human lung fragments, in parallel to guinea pig lung, released PGE, PGF2alpha and the metabolites when incubated with SRS or antigen. In in vivo experiments, SRS and arachidonic acid given intravenously increased the airway insufflation pressure in anesthetized quinea pigs. These effects, but not the action of injected PGF2alpha and histamine, were abolished by indomethacin. The results indicate that one of the modes of SRS action is by release of PGs, and are consistent with the hypothesis that PGs are predominantly "secondary" mediators (in the temporal sense) of the antigen-antibody reaction.


Dynamic and structural features of a case of compulsive neurosis as revealed in dreams. Several examples of the manifest dreams reported by a patient with a compulsive neurosis are presented. An outstanding feature of the dreams was the sudden shift of the dreamer's self-representation from the role of criminal to the role of policeman. This shift is thought to parallel the oscillation between unconscious instinctual gratification and conscious attempts at reparation which is the main dynamic feature of the compulsive neurosis in waking life. The shift of orientation of the self reflects the operation of early defense mechanisms against the sadistic impulse. It also portrays graphically an early stage in the internalization process which leads ultimately to the creation of the superego. The repetitive nature of the dreams indicated that the patient had become fixated at this particular point in the course of his psychic development. It is interesting to note that like the Rat Man this patient suffered from a chronically undescended testicle. The real defect in the sexual apparatus in both patients may have been a factor which favored regression of the libido from the genital stage to the anal-sadistic stage.


[Mental illness and stress].

[Mental illness and stress]. Main purpose of my presentation was to give a review on the effects of the stress-factors on the psyche. This aspect has not been taken into consideration sufficiently yet in theory of stress. I like to stress once more that it is the excessive, too intensive stress reaction which is leading to psychic disturbances as well as functional disturbances. Stress which does not exceed a certain extent has to be considered physiologically: the organism needs it as stimulans to obtain its functions: it could not survive without this stimulans. The situation only becomes risky if the stress factors are causing pathogenous disturbances. This situation has been named "dis-stress" by Selye. The best guarantees to avoid "dis-stress" as well as psychic stress-disorders are changing of attitude of life, compensation of overstrain by sensible recreational activities, going in for sport, engagement for human and social environmental factors as well as a harmonic family-life and a conception of life in the sense of imperturbability and tolerance.


Control of extracellular slime accumulation in monokaryons and resultant dikaryons of Schizophyllum commune. Extracellular slime accumulation, as alcohol-precipitable material was measured after eight days of growth in glucose-asparagine-salts broth in twenty-two different monokaryons and six resultant dikaryons of Schizophyllum commune. The nutritional control of slime accumulation was also examined in monokaryotic mycelium. Slime occurred after growth in sucrose, glucose, fructose and xylose, with glycerol best. Low inorganic phosphates limited both slime and mycelial growth while limiting MgSO4 decreased growth and enhanced slime. In glucose-asparagine broth, various monokaryons differed widely in slime accumulation, ranging from none (e.g., strain 19) to nearly 800 mg per 100 ml filtrate (strain 1) after eight days growth, followed by a marked decline in slime (eleven days to twenty-one days). Resultant dikaryons all showed less slime accumulation, even when established from two high slime-accumulating monokaryons. In contrast, conditions which arrested dikaryotic fruit-body morphogenesis led to increased slime accumulation.


Lipolytic enzymes of Trichophyton rubrum. Trichophyton rubrum cells contain lipase, phospholipases A and B and acyl CoA lysolecithin acyl transferase activities. This dermatophyte excretes lipase and phospholipase A into the growth medium when cultivated in Sabouraud's broth. Extracellular lipase has optimum activity at pH 8.0 whereas the intracellular lipase is maximally active at pH 8.0 whereas the intracellular lipase is maximally active at pH 7.0. The optimum pH of phospholipase A and B activities which are localized in 15000 g sedimentable cell fragments are 7.0 and 6.0 respectively. Supernatant obtained after removal of 1,005,000 g sedimentable fragments from cell extract contains acyl CoA lysolecithin acyl transferase which requires ATP, CoA, Mg2+ and an unsaturated fatty acid for its activity.


[The prevention of accidents in dental local anesthesia]. The relatively rare complications developing under local anaesthesia may be due to various causes. The possibilities of the development of accidents under local anaesthesia are outlined; and prophylactic and therapeutic measures are deduced. A schematic premedication cannot prevent the development of complications with a sufficient degree of certainty; for instance, in certain cases the administration of atropine alone will still worsen the course of a complication. On the contrary, an individual premedication should be aimed at, especially in risk patients; the dosage must take into consideration all important factors. Consequently, atropine is of less importance than, for example, psychopharmaca and beta-receptor blockers.


Uric acid calculi: results of conservative treatment. A total of 140 patients with uric acid diathesis have been treated for the last five years with a standardized mixture of sodium and potassium citrate and citric acid (Uralyt U) together with forced fluid intake and, if necessary, the addition of allopurinol. The results have been encouraging; 80 per cent of the patients showed complete dissolution of the stones and relief of symptoms.


Endocrine tests in phenotypic children with bilateral impalpable testes can reliably predict "congenital" anorchism. Congenital anorchism is a rare condition. Bilateral impa-pable undescended testes are relatively common by comparison. Surgical exploration has been regarded as the final arbiter between anorchism and bilateral cryptorchism. Exploration has not proved completely reliable in making this differentiation. Endocrine studies, particularly the human chorionic gonadotropin (HCG) stimulation test together with measurements of basal plasma gonadotropins, can reliably exclude "functioning" testicular tissue. Eleven fully evaluated and operated cases support this contention. In the specific clinical setting of a normal phenotypic male child with a 46XY karyotype and no müllerian structures palpable on rectal examination, nonfunctioning testes on endocrine testing means congenital anorchism and surgical confirmation is unnecessary. In contradistinction, a positive HCG test would appear to mandate through and extensive surgical exploration.


[Acid-base indices in ovine blood and cerebrospinal fluid]. By means of the Astrup apparatus (Blood Microsystem BMS 2) acid-base indices in blood and cerebro-spinal fluid of clinically healthy Merino sheep at the age of one year (15 head) and of two or three years (41 head) were determined. Blood collected anaerobically from v. jugularis and liquor collected by means of the suboccipital punction in heparinized syringes and/or caplaris and liquor collected by means of the suboccipital punction in heparinized syringes and/or capillaries were examined immediately after the collection. Following indices were obtained and re-counted according to nomograms: actual pH, partial CO2 pressure--pCO2 base excess--BE, base buffer--BB, standard bicarbonate--SB, actual bicarbonate--AB, total CO2--tCO2 (Tables I, II). Indices of pH, pCO2, BE and BB indicated statistically significant differences in values when comparing liquor with venous blood (in younger ewes). As the values of the sheep cerebro-spinal fluid indices examined are not to be found in literature, the above stated values can be recommended to be used as basic values for a given age group.


[Rat brain and liver glutamine synthetase and gamma-glutamyltransferase activity in alloxan diabetes (IV)]. Glutamine synthetase and gamma-glutamyltransferase activities of brain and liver homogenates of rats suffering from alloxan diabetes were determined in the soluble fraction (fraction 1) and in that obtained after treatment with 0.2 percent deoxycholate (fraction 2). The results obtained indicate that the activities of these enzymes in homogenates of brain and liver of diabetic animals does not differ from that of normal animals. gamma-Glutamyltransferase activity of brain is significantly reduced (about 5 fold) in the soluble fraction while glutamine synthetase activity is not much changed. The activities of glutamine-synthetase and gamma-glutamyltransferase of the 2-nd fraction obtained from rat brain and liver are very much higher than in the first fraction and are not considerably different from the activities observed in normal animals. In contrast to brain, glutamine synthetase and gamma-glutamyltransferase activities of liver of diabetic animals do not differ from the activities observed in normal animals, both in the homogenates and in the 1-st and 2-nd fractions.


[Mechanisms regulating citric acid metabolism in the brain]. The changes in the rates of citrate biosynthesis and utilization in rat brain, liver, kidney and heart, produced by hypoxia, action of 2,4-DNP and thyreotoxicosis, were compared with changes of some regulatory parameters under the same conditions. The comparison of citrate-synthase activities, citrate levels in tissues and 14C-incorporation from different precursors into citric acid permitted us to establish that the biosynthesis of citrate in brain was more intensive than in other tissues studied. The main source of acetyl-CoA for citrate-synthase reaction in brain is the oxidation of pyruvate. The ratio of adenine nucleotides plays an important role in the control of citrate-synthase activity in brain, where the oxaloacetate control is not as significant as in liver. NAD-specific isocitrate dehydrogenase reaction was found to be the dominant pathway for citrate oxidation in brain: more than 60 percent of brain citrate were oxidized by NAD-ICDH, while less than 10 percent of citric acid were utilized by this enzyme in other tissues studied. The existance of an adenine nucleotide control of NAD-ICDH activity in brain may be an additional mechanism for the regulation of the first steps of energy metabolism in brain.


Comparative metabolism of benorylate and an equivalent mixture of aspirin and paracetamol in neonate and adult rabbits. 1. Benorylate was well absorbed in rabbits, but more slowly than an equimolar mixture of aspirin and paracetamol. 2. Benorylate was extensively hydrolysed and converted to the typical metabolites of aspirin and paracetamol by both neonate and mature rabbits. 3. Absorption of either aspirin-paracetamol or benorylate was slower in neonate rabbits than in adult rabbits. 4. The excretion rate in adult rabbits was faster, for both aspirin and paracetamol metabolites, than in neonate rabbits.


Orofacial dyskinesia. Clinical features, mechanisms and drug therapy. Orofacial or tardive dyskinesias are involuntary repetitive movements of the mouth and face. In most cases, they occur in older psychotic patients who are in institutions and in whom long-term treatment with antipsychotic drugs of the phenothiazine and butyrophenone groups is being carried out. These dyskinesias are frequent in occurrence and characteristically are irreversible. Several biochemical mechanisms have been proposed as causes, including hypersensitivity or partially deneverated brain dopamine receptors and low affinity of the offending drugs for brain muscarinic cholinergic receptors. Clinical therapy has been attempted primarily with drugs that antagonize dopamine receptors or deplete brain dopamine. The benefits of drug treatment have been variable and lack of consistent improvement has been discouraging. Early recognition of dyskinesia should be attempted, and the dose reduced or the drug omitted at the first sign.


[Metabolic acidosis in chronic renal insufficiency of various etiology]. The renal acidosis may appear by a decrease of the number of nephrons able to function (decrease of the filtrate of the glomerulum) as well as by a selective alteration of the tubular acidification mechanism. In 66 patients with chronic renal insufficiency of different degrees of severity (32 patients with diabetic glomerulosclerosis, 18 patients with chronic glomerulonephritis and 16 patients with chronic pyelonephritis) studies of the parameters of the acid-base-state and the renal insufficiency were carried out. 53 of these patients had a pathologically changed acid-base-state which was most expressed in patients with chronic pyelonephritis. A characteristic relation between the renal function (creatinine clearance) and the change of the pH-values was observed. The regression curve of the pH-values was descending so that in clearance values below 25 ml/min in nearly all patients a pronounced acidosis was present. This could be proved in the patients with pyelonephritis already when higher clearance values were present. The forms of the development of the metabolic acidosis in chronic renal insufficiency are discussed.


[Use of the 25-point ocular grid plate in quantitative-morphological studies of eutopic and dystopic testes of children]. From plains histologic preparations of children's testicles the volume of seminiferous tubules and extratubular tissue were calculated by use of the points counting method. In ectopic testicles, the extratubular part is significantly larger than in normally descended testes. The range of the values measured by this method diminishes the higher the total count of test points is. The 25-points-test-net of VEB Carl Zeiss Jena in particular met all requirements for quantitative examinations of the morphology of testicles.


[Consideration of the mesenchyme-reaction in patients with cronical liver-diseases (author's transl)]. The lysosomal glycosidase N-Acetyl-beta-glucosaminidase (beta-NAG) is involved in the breakdown of connective tissue. The activity of this enzyme was determined in sera and biopsies of liver tissue from patients with chronic diseases of the liver of different aetiology. Elevation of enzyme activity was found to be related to an increase of the mesenchymeraction in the injured liver tissue. No correlation was found between activity of beta-NAG and of GPT, gammaGT and CHE.


gamma-Radiolyses of DNA in oxygenated aqueous solution. Structure of an alkali-labile site. Erythritol-1-d1 has been isolated from gamma-irradiated aqueous oxygenated solution of DNA after reductions with NaBD4, alkali and phosphatase treatment. It is concluded that this product stems from a D-erythrose 2,4-diphosphate unit in the DNA which is formed via a sequence of reactions following H-abstraction at C-2' by OH radicals.


Effect of ligands of ferric hemes on interaction between ferric and ferrous chains in partially oxidized hemoglobin A. Normal values of Bohr effect of oxygenation of partially oxidized hemoglobin A with ferrihemes liganded either with H2O and OH or with CN have been found in the range of pH values from 6.8 to 7.6 in 45 micrometer (Fe)-hemoglobin containing 36--38% of ferrihemes. As the changes of oxygen affinity of Hb A induced by changes of pH are due to the modifications of R state, this quaternary conformation is thought to be unchanged in the studied of R state, this quaternary conformation is thought to be unchanged in the studied forms of partially oxidized hemoglobin. It is suggested that interactions between ferric and ferrous hemes leading to the increased affinity of ferrous hemes to oxygen occur in deoxygenated form of partially oxidized hemoglobin. In partially oxidized hemoglobin with ferric hemes liganded with H2O asymmetry of oxygen binding curves has been noted, which is not observed in forms with ferric hemes liganded with OH ot CN. This shows the effect of ligands of ferric hemes on interactions between chains containing ferric and ferrous hemes.


[The effect of thyroxine on the 2,3-diphosphoglycerate content of erythrocytes in vivo and in vitro]. After ending a continous treatment with thyroxine the average dropping of the 2,3 DPG level was 0.4 mumol/ml. T4 decreased on the average by 7.6 microgram/ml. One time application of 1 mg thyroxine p.o. led within 24 hours to an increase of the 2,3 DPG level of -chi = 0.2 mumol/ml, the pH in the erythrocytes increased by 0.02 on the average. Blood incubation with thyroxine added in a concentration of -chi = 24 microgram/100 ml showed no increase of 2,3 DPG, pH and phosphate, while there was a significant acidosis and increase of phosphate in the control blood. The lactate production was significantly lower and glucose consumption was significantly higher in the blood with thyroxine.


Characteristics of a new abnormal variant of G-6-PD in human red cells. Kinetic and electrophoretic properties were studied in 230--300 fold purified preparations of glucose-6-phosphate dehydrogenase (G-6-PD) from red cells of donors and patients with hemolytic anemia induced by G-6-PD deficiency. In abnormal variant of G-6-PD isolated from red cells of a patient with hemolytic anemia which had not before been described in the literature was found. The abnormal variant differs from the normal enzyme by a decreased Michaelis constant for G-6-P and NADP, by increased utilization of substrate-analogues (2-deoxy-G-6-P and deamino NADP in particular), by low heat stability, the character of pH dependence, and by the appearance of one band of G-6-PD activity during electrophoresis in polyacrylamide gel. The isolated abnormal variant of G-6-PD has been called "Kremenchug" according to the origin of the patient.


Regulation of NAD and NADP synthesis in human red cell. NAD is synthesized in red cell from nicotinic acid and PRPP through the formation of nicotinate mononucleotide and desamido-NAD. Synthesis of one mole of NAD requires two moles of ATP. NADP comes from NAD phosphorylation by NAD-kinase (EC.2.7.1.23). NAD and NADP analysis on a population with ATP level ranging from 800 to 2500 nmoles/ml red cells showed a close correlation between ATP and pyridine cofactors. Moreover, NADP level appeared to be dependent of the redox-state of NADP/NADPH couple. Subjects with low NADPH (G-6-PD) deficient red cells, Hb Köln) showed lower NADtot/NADPtot ratio, suggesting a NAD-kinase equilibrium shift toward NADP related to lower levels of the negative effector NADPH, as already described in rat liver.


[Glucose-6-phosphate dehydrogenase deficiency of erythrocytes in the GDR]. 34 persons with G-6-PD deficiency were diagnosed, and the pathological enzyme-variants of red blood cells were characterized according to the recommendations of WHO. We conclude from the differing residual G-6-PD-activities in red blood cells of the propositi and the differing reactivity of the enzyme in kinetic and physicochemical characterizations that a multiple variety of rare pathological G-6-PD variants exists in the GDR. Using the estimated enzymeparameters it was not possible in all cases to compare directly the newly demonstrated G-6-PD variants with cases already described in the literature. In addition, the differing combinations of parameters render a classification more difficult.


[Detection of female heterozygous glucose-6-phosphate dehydrogenase deficiency]. Diagnostics of heterozygotes are required for population studies, for the detection and consultation of persons with G-6-PD deficiency prone to hemolysis. The diagnostics of heterozygous females with the corresponding trait are problematic in families without hemizygous patients. 1. The determination of the activity is only applicable to the differentiation between heterozygotes and homozygotes if the activities are below the reference range. Heterozygous G-6-PD deficiency with normal activity cannot be identified by this method. 2. Existence of G-6-PD defects is demonstrated by mosaicism even in case of normactivity (Tönztest). 3. Incubation with and without NADP of stroma-free hemolysates involving heat labile enzyme mutants results in a marked decrease of activity within 20 min at 46 degrees C. 4. Electrophoresis on Cellogel demonstrates changes of charge in the mutated enzyme. 5. Family examination verifies suspicion of the heterozygous trait. A combination of parameters is recommended to obtain an improvement in the detection of persons with the heterozygous trait.


Mechanism of senescence of red blood cells. An integral hypothesis is submitted on the interaction of endogenous and exogenous factors in the mechanism of senescence of erythrocytes and on their selective phagocytosis by autologous macrophages. A method that allows quantitative determination and preparative yield of senescent erythrocytes is proposed.


[Sedimentation rate of erythrocytes as an indicator for phase transitions in the membrane]. Temperature dependent sedimentation measurements with human erythrocytes showed that the sedimentation velocity at 21 degrees C in Krebs-Ringer-solution, pH 7.4, has a peak shaped minimum [1]. In further investigations it could be shown that a phase transition of membrane lipids is the main reason of this peak. Membrane proteins influence the sedimentation behavior of the erythrocytes only by changing the interaction with membrane lipids.


Serum gamma-glutamyl transpeptidase: its clinical significance. Serum gamma-glutamyl transpeptidase (gamma-GT) level was estimated in 132 patients with different liver diseases (chronic persistent and chronic active hepatitis, postnecrotic cirrhosis, chronic alcholic hepatitis and alcoholic cirrhosis, cholestasis syndrome, fatty liver, Gilbert disease) and malignancies with and without liver involvement. The gamma-GT levels were compared with the values for serum bilirubin, transaminases (GOT, GPT) and alkaline phosphatase in the same patients. gamma-GT values were normal in chronic persistent hepatitis and increased in chronic active hepatitis. Very high activities were measured in chronic alcoholic cirrhosis in contrast to postnecrotic cirrhosis. gamma-GT proved to be more sensitive than alkaline phosphate as an index of cholestasis and liver involvement in malignancies. It is suggested that gamma-GT activity offers valuable aid in differential diagnostics of liver-diseases. gamma-GT being an inducible enzyme, its activity may be raised by enzyme inducing drugs also in subjects without liver disease.


[Painful ophthalmoplegia (author's transl)]. The authors begin by enumerating the various syndromes in which painful ophthalmoplegia may be observed (sphenoidal fissure syndrome, Collier's syndrome, syndromes involving the orbital apex, the cavernous sinus and parasellar syndromes; Raeder's syndrome, Gradenigo's syndrome and Fischer-Brugge syndrome). They then discuss the various causes that must be investigated in all cases of painful ophtalmoplegia. They consider in order: -- ophtalmoplegia due to general causes (especially diabetes) and neurological causes (e.g. multiple sclerosis); -- ophtalmoplegia due to common local canses space-occupying processes, vascular malformations, ear, nose and larynx infections); -- painful ophalmoplegia of unknown origin, which includes four entities of very differing importance (Gubler and Charcot's ophthalmoplegic migraine and Tolosa-Hunt syndrome of which the clinical symptoms and course are so different that they can be distinguished as two entities; and, secondarily, inflammatory pseudo-tumours of the orbit and the recurrent multiple cranial nerve palsies that are observed in South-East Asia).  (Acta nurol. belg., 1977, 77, 331-350).


Naphthylamidase used as a lysosome marker in the study of acute selective necrosis of the internal granular layer of cerebellum. Histochemical examination of the activity of naphthylamidase (LNAse) in the cerebellar cortex of 70 human autopsies consistantly revealed a marked activity mainly in the internal granular layer with pH optimum of 5.8. Slight enzyme activity was also localized in sites corresponding to lipofuscin deposits and areas of acid phosphatase activity in the Bergmann glial cells, Purkinje cells and in perivascular cells. The histochemical findings support the LNAse reaction as a lysosome marker. Differences in localization of LNAse and acid phosphatase could possibly be due to prior release of the latter enzyme from the internal granular layer. Significant correlation between demonstrable loss of granule cell nuclei (the so-called acute, selective necrosis of the granular layer) and low pH of the cerebellar tissue could be demonstrated in 21 cases. The present findings support the hypothesis that an enzymatic disintegration of the granule cells takes place in postmortem cerebella with low pH simulating a necrotic vital phenomenon.


Surface-charge characteristics of smooth and rough Salmonella typhimurium bacteria determined by aqueous two-phase partitioning and free zones electrophoresis. Aqueous biphasic partitioning of Salmonella typhimurium S and R bacteria in a system containing 6.2 per cent (w/w) dextran 500 and 4.4 per cent (w/w) poly(ethyleneglycol) 6000 (PEG) was similar to the partition of the corresponding surface lipopolysaccharide (LPS). Further partition analysis with charged PEG showed that S bacteria and their LPS exposed very little charge, whereas R bacteria and their LPS showed a conspicuous negative charge at neutral pH. Free zone electrophoresis also indicated that the S bacteria have a much lower surface charge density than the R bacteria and accordingly a different surface structure. Thus, the physico-chemical properties of the bacterial surface seem to be determined to a great extent by the characteristics of the cell surface LPS.


Physiological and pathophysiological aspects of gastrointestinal peptide hormones. The author made a review about the origin, the biochemistry the physiological and pathological roles of gastrointestinal peptide hormones. They originate from the APUD cell system, chemically from the ancient growth hormone, or placental lactogen. The theoretical prosecgastrin's first sequencies form the "secretin family", the tail sequencies form the "gastrin family". The author describes many details of their effects on the different gastrointestinal organs, they behave mainly antagonistic way to each other. Finally a discussions is given about their role in the development of peptic ulcer, in the WDHA syndrome and in malabsorption.


Maintenance of Glossina palpalis fed through bat's wing membrane on defibrinated blood. Teneral laboratory-bred Glossina palpalis flies were successfully fed through bat's wing membrane on defibrinated blood by means of a technique developed in this laboratory. Using this technique almost 100% of the flies engorged blood through the membrane. Ten to fifteen flies were kept in Standard 'Geigy 10' or 'Geigy 15' cages and records were kept of mortality and the weights of the pupae produced; over 78% of the flies were still after 60 days of the experiment. Over 75% of the membrane-fed G. palpalis females produced pupae in the first 30 days of the females' reproductive life. The mean weight of the pupae produced by the membrane-fed flies was 24.9 mg. The results indicate that membranes made from the African fruit bat wings are usable, and that this type of membrane would be valuable for a medium scale rearing programme for tsetse flies.


[The etiology of sylvatic trichinellosis in Switzerland]. Three Trichinella strains from Swiss wild foxes (Vulpes vulpes) were tested in crossbreeding experiments with Trichinella spiralis, T. nativa and T. nelsoni using white mice as laboratory animals. The results demonstrated that T. nelsoni Britov et Boev 1972 is the etiologic agent of sylvatic trichinellosis in Switzerland. Former isolations of these species were successful in materials from East and South Africa, USSR (Republics of Tadzhikistan, Turkmenia and Kazakhstan) and Bulgaria. The area of T. nelsoni covers probably the whole southern part of the Old World up to the 48th degree of northern latitude.