Determination of deuterium-labeled tryptophan in proteins by means of high-performance liquid chromatography and thermospray mass spectrometry

In order to develop direct methods for determining the extent of metabolic incorporation of isotopically labeled amino acids into a protein, the determination of deuterated tryptophan in [2H5]tryptophan-bacteriorhodopsin was investigated. The isotopically modified protein was subjected to alkaline hydrolysis. After phenyl isothiocyanate derivatization of the hydrolysate, the mixture was separated by reversed-phase liquid chromatography. Field desorption mass spectrometry and thermospray mass spectrometry were investigated for their ability to determine the ratio between [2H5]tryptophan and total tryptophan in the collected fractions. In order to check the procedure a set of known tryptophan/[2H5]tryptophan mixtures were passed through the same derivatization, HPLC separation, and lyophilization procedure as used for the biological samples.     

Determination of the structural integrity of DNA from pathogenic enterobacteria

Among the enterobacterial strains under study, more organisms in the stationary phase of growth have been found to have nicks in their DNA than those in the exponential phase. Bacteria less sensitive to ultraviolet irradiation have the least number of nicks in each phase of growth. The number of nicks in different strains belonging to the serovar is sufficiently stable. Virulent and avirulent forms show no difference in this characteristic.     

Cl- transport in apical plasma membrane vesicles isolated from bovine tracheal epithelium

Gradually altered synthetic entities were employed as molecular probes, and arachidonic acid, ADP, human alpha-thrombin and the Ca2+ ionophore A23187 as aggregation-inducing agents, in a comprehensive study on the response profile of human blood platelets with an emphasis on the effects of exogenous and increased intracellular Ca2+. Corroborating further previous conclusions, some representative carbamoylpiperidine derivatives, at concentrations effecting substantial inhibition of ADP-induced aggregation, failed to retain that effect when 5.0 mM Ca2+ was introduced into the otherwise identical test medium; reference compounds chlorpromazine and propranolol registered corresponding inhibitory patterns. At increased concentrations the compounds' inhibitory potency was regenerated even in the presence of 5 mM Ca2+. In fact, in sufficiently high concentrations, the compounds were even capable of inhibiting aggregation elicited by 15 microM of the ionophore A23187; so did chlorpromazine and propranolol. Another set of congeners revealed the striking sensitivity of ionophore A23187-induced human blood platelet aggregation to the surface active potencies of inhibitor molecules. The loss in inhibitory potency was directly related to the lesser hydrophobic character of the molecule.     

Regulation of glutaminase by exogenous glutamate,ammonia and 2-oxoglutarate in synaptosomal enriched preparation from rat brain

Phosphate activated glutaminase in synaptosomal enriched preparation from rat brain is very sensitive to inhibition by low concentration of glutamate, ammonia and 2-oxoglutarate when added to the incubation medium at pH 7.6. By increasing the concentration of either of these compounds up to 0.5 mM a pronounced initial inhibition is followed by little or no further effect when the concentration is increased beyond this level. By lowering the pH of the reaction mixture to 7.0, the inhibition by glutamate is almost abolished and that of ammonia reduced. Glutamate inhibits mainly the N-ethylmaleimide-sensitive fraction of glutaminase which previously is suggested to be localized to the outer phase of the mitochondrial inner membrane, whereas ammonia inhibits both the N-ethylmaleimidesensitive and-insensitive fraction. Evidence has been produced to show that the inhibition by 2-oxoglutarate is caused by glutamate formation by aminotransferase reactions. Since 2-oxoglutarate is produced by the tricarboxylic acid cycle, the operation of this cycle may regulate the glutaminase reaction by controlling glutamate formation via the aminotransferase reactions.Abbreviations used NEM N-ethylmaleimide - PAG phosphate activated glutaminase - AOA aminooxyacetic acid     

On tannic acid fixation and staining

Tannic acid was found to fix and stain glycocalyx heavily. After removal of the major component of surface glycopeptides by trypsin, the surface coat was stained vaguely, and after the treatment with collagenase, the surface coat was moderately stained. It is concluded that tannic acid stained non-specific surface glycopeptides.     

Zur Taxonomie von Rhodopseudomonas palustris

Zusammenfassung Eine Reihe von Rhodopseudomonas palustris-Stämmen aus verschiedenen Herkünften wurden vergleichend unter Verwendung folgender Merkmale untersucht: Substratverwertung, in vivo-Absorptionsspektrum und Serologie der O-Antigene. Die gegen 2 Stämme gerichteten Antiseren zeigen hohe Spezifität. Die Verwendbarkeit der serologischen Kreuzreaktion für taxonomische Untersuchungen bei photosynthetischen Bakterien wird diskutiert.

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On the taxonomy of Rhodopseudomonas palustris

Summary Strains of Rhodopseudomonas palustris isolated from different habitats were compared with respect to their taxonomic features. All strains grew very well on formiate, acetate, propionate, butyrate, aspartate, inositol, ethanol, fructose, and p-amino-benzoate, respectively, as single carbon source. Most of the strains were able to use benzoic acid or glucose, too. But alanine was not found to be a good substrate. The maxima of the bacteriochlorophyll in-vivo-absorption spectra were estimated to be 376, 589, 802–805, and 858–875 nm. The shift of the infrared peak in the different strains is loosely correlated with the change of the carotenoid in vivo spectrum, the maxima of which were measured to be 470–480 nm (shoulder) 495–505 nm, and 520–545 nm (shoulder). Antisera were prepared against the strains 1e5 and 11/1. It was demonstrated that these antisera were directed against the lipopolysaccharides (O-antigen) of these bacteria. The antigen of 1e5 does not cross react with the antigen of 11/1. Strain 1e5 is the only one of 17 strains tested which is sensitive to the bacteriophage Rp1. The antigen of this strain cross reacted only with the antigen of strain K1. In contrast, the antigen of strain 11/1 cross reacted in some degree with most of the tested strains of Rps. palustris. No or very weak cross reaction was observed between the antigens of Rps. palustris (1e5, 11/1) and Rps. capsulata, Rps. spheroides, or R. rubrum, respectively. In contrast to 11/1 only heat-killed cells of strain 1e5 were agglutinated by anti-1e5.

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Im Text verwendete Abkürzungen LPS Lipopolysaccharid - R Rhodospirillum - Rps. Rhodopseudomonas - i.m. intramuskulär - s.c. subcutan - i.v. intravenös     

Reactivity of neurons of the orbito-frontal cortex of the cat during exposure to the scent or sight of food

Reactivity of neurones in orbitofrontal cortex of the cat to the action of light or sound was studied in consecutive stages of alimentary behaviour conditioned by the smell and then the sight of food. Changes were found of the character of neuronal reactions to the light and sound stimuli at the change of smell to the sight of food. A conclusion is drown, that polysensory properties of the neurones of the orbitofrontal cortex provide integral organization of brain sensory function at separate stages of alimentary behaviour.