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Screening of thiol compounds: depolarization-induced release of glutathione and cysteine from rat brain slices.

Superfusates from rat brain slices were screened for thiol compounds after derivatization with monobromobimane by reversed-phase HPLC. Only glutathione and cysteine were detected. The Ca(2+)-dependent release of these compounds from slices of different regions of rat brain was investigated, applying a highly sensitive and reproducible quantification method, based on reduction of superfusates with dithiothreitol, reaction of thiols with iodoacetic acid, precolumn derivatization with o-phthalaldehyde reagent solution, and analysis with reversed-phase HPLC. This methodology allowed determination of reduced and total thiols in aliquots of the same superfusates. Mostly reduced glutathione and cysteine were released upon K+ depolarization and the Ca2+ dependency suggests that they originate from a neuronal compartment. The GSH release was most prominent in the mesodiencephalon, cortex, hippocampus, and striatum and lowest in the pons-medulla and cerebellum. This underscores a physiologically significant role for glutathione in CNS neurotransmission.

Animals↗

Killing of cells in bacterial colonies.

Bacterial colonies grown on membrane filters and transferred to plates of inhibitor-containing medium decrease in number of total viable colony-forming cells. The decrease in viable cell count occurs in direct proportion to the concentration of inhibitor present and the length of time the colonies are exposed to the inhibitor, suggesting that the kinetics of cell kill approximate a first-order chemical reaction.

Anti-Bacterial Agents↗

Effect of lowered intracellular ATP and GTP concentrations on purine ribonucleotide synthesis and interconversion.

The effects of lowered intracellular ATP and GTP concentrations on enzymes of purine ribonucleotide synthesis and intercoversion were studied using intact Ehrlich ascites tumor cells. The apparent rates of phosphoribosyl pyrophosphate synthetase (EC 2.7.6.1) and of inosinate dehydrogenase (EC 1.2.1.14) were increased in cells containing lowered purine nucleotide concentrations, but apparent activities of amidophosphoribosyltransferase (EC 2.4.2.14), the purine phosphoribosyltransferases, and other enzymes of purine ribonucleotide interconversion were not affected.

Adenine Phosphoribosyltransferase↗

Hypoxanthine-guanine phosphoribosyltransferase variant associated with accelerated purine synthesis.

We have previously described a 14-yr-old boy with hyperuricemia, renal failure, and accelerated purine production resistant in vivo and in vitro to purine analogs. This patient demonstrated normal red cell hypoxanthine-guanine phosphoribosyltransferase (HPRT) heat stability, electrophoresis at high pH, and activity at standard substrate levels. In the present report an abnormal HPRT enzyme was demonstrated by enzyme kinetic study with phosphoribosylpyrophosphate (PRPP) as the variable substrate and inhibitory studies with sodium fluoride. Apparently normal HPRT activity in a patient with hyperuricemia and gout does not exclude a functionally significant HPRT mutation.

Adolescent↗

Trypsinized BHK21 cells aggregate in the presence of metabolic inhibitors and in the absence of divalent cations.

The rapid formation of adhesions in suspension by lightly trypsinized BHK21 cells is not dependent on protein synthesis, and only in part on cellular metabolism, although it is completely inhibited by heat- and aldehyde-fixation of the cells. A requirement for protein synthesis becomes evident only if cells are exposed to high levels of trypsin for long periods. Formation of adhesions does not require addition to the medium of divalent cations, although it is increased by divalent manganese and cobalt ions. It is promoted by cytochalasin B and by cyclic AMP and is not inhibited by p-mercuriphenylsulphonate. We discuss a possible relationship between aggregation and the formation of gap junctions.

4-Chloromercuribenzenesulfonate↗

Hypertrophic foci of pancreatic acinar cells in rats.

A morphologically distinctive type of pancreatic acinar cell foci, different from hyperplastic nodules and adenomas, in rats has been recognized for two decades. The lesions have been observed to occur spontaneously and to be induced experimentally. They consist of enlarged acinar cells with abundant cytoplasm of altered staining characteristics and prominent nuclei. There is, however, a wide divergence of opinion among investigators regarding the nature of the lesions. As a result of different interpretations and classifications, many terms have been given to them. Based on morphologic characteristics, the author has designated the lesions as hypertrophic foci, a descriptive morphological term. The biologic significance with particular reference to age and the relationship with acinar cell neoplasia is discussed. Also included in the review are similar lesions in other rodent species.

2-Acetylaminofluorene↗

Regulation of de novo purine biosynthesis in human lymphoblasts. Coordinate control of proximal (rate-determining) steps and the inosinic acid branch point.

Purine nucleotide synthesis de novo has been studied in a permanent tissue culture line of human splenic lymphoblasts with particular attention to coordination of control of the proximal (rate-determining) steps with the distal branch point of the pathway. An assay was used which permits simultaneous determination of the overall rate of labeling of all intracellular purines with sodium [14C]formate, as well as the distribution of isotope into all intracellular guanine- and adenine-containing compounds. The guanine to adenine labeling ratio was used as an index of IMP branch point regulation. It was found that exogenous adenine and guanine produce feedback-controlling effects not only on the first step in the de novo pathway, but also on the IMP branch point. Concentrations of adenine which produce less than 40% inhibition of the overall rate of de novo purine synthesis do so by selectively inhibiting adenine nucleotide synthesis de novo by 50 to 70% while stimulating guanine nucleotide synthesis de novo by up to 20%. A reciprocal effect is seen with exogenous guanine. The adenosine analog 6-methylmercaptopurine ribonucleoside selectivity inhibits adenine nucleotide synthesis via the de novo pathway but not from exogenous hypoxanthine. Thus, the reactions of purine nucleotide interconversion, in particular adenylosuccinate synthetase, may be regulated differently in cells deriving their purine nucleotides solely from de novo synthesis than when deriving them via "salvage" of preformed hypoxanthine.

Azaserine↗