Multiple components in an epidermal growth factor-stimulated protein kinase cascade. In vitro activation of a myelin basic protein/microtubule-associated protein 2 kinase.

Epidermal growth factor stimulates the activity of several cytosolic serine/threonine protein kinases in quiescent Swiss 3T3 cells. Two of these, which use myelin basic protein (MBP) as substrate, act as kinase kinases in that they are able to activate a separate peptide kinase activity in vitro by a mechanism involving protein phosphorylation. In this study, we have identified two activities from extracts of epidermal growth factor-treated cells that stimulate an ATP-dependent activation of both of the MBP kinases, derived in their inactive precursor forms from extracts of untreated cells. The resulting MBP kinase activities are stable to further purification and can be inactivated with either tyrosine or serine/threonine protein phosphatases and then reactivated to their original levels of activity. Thus, we propose that the in vitro activation involves protein phosphorylation, stimulated by the action of novel MBP kinase activating factors that represent intermediate components in a growth factor-stimulated kinase cascade.     

Kinetic properties of a L-cysteine desulfhydrase-deficient mutant in the enzymatic formation of L-cysteine from D,L-ATC

Summary A mutant strain lacking in activity of L-cysteine desulfhydrase, a L-cysteine-decomposing enzyme, was screened after UV-treatment ofPseudomonas sp. CU6. The properties of the two strains, original and mutant, were compared on the basis of parameter values estimated from kinetic simulations of the enzymatic formation of L-cysteine from D,L-ATC. Both strains suffered from product inhibition, though inhibition was less for the mutant strain.     

Expression of glucoamylase gene usingSUC2 promoter inSaccharomyces cerevisiae

Summary The cloning of glucoamylase geneSTA using theSUC2 promoter intoSaccharomyces cerevisiae was performed. The signal sequence ofSTA gene was used for the secretion of glucoamylase protein. The plasmid constructed in this way was named YEpSUCSTA and its expression was identified. The expression of YEpSUCSTA was repressed in the presence of glucose in growth medium, but derepressed when glucose became depleted. YEpSUCSTA showed the similar efficiency of glucoamylase secretion as YEpSTA-F which has the entireSTA gene. Glucoamylase activity in starch-glucose medium was largely increased because cell mass and plasmid stability were high in biosynthesis phase compared to extracellular glucoamylase activities in media which starch or glucose was the only carbon source.     

Ammonium ingestion prevents depletion of hepatic energy metabolites induced by acute ammonium intoxication

Ingestion of an ammonium containing diet produces hyperammonemia and protects rats against acute ammonium intoxication. Acute ammonium toxicity has been attributed to the depletion of energy metabolite intermediates. We show here that hyperammonemia affords considerable protection against depletion of hepatic energy metabolites evoked by ammonium acetate injection. In control rats there were marked decreases in the content of acetoacetate, beta-hydroxybutyrate, ATP, 2-oxoglutarate, lactate, and pyruvate while phosphoenolpyruvate increased markedly. In hyperammonemic rats beta-hydroxybutyrate, ATP, 2-oxoglutarate, and lactate were not significantly affected while pyruvate increased markedly and phosphoenolpyruvate slightly. These results suggest that in controls the activity of pyruvate kinase is inhibited after ammonium injection while in hyperammonemic rats it is not inhibited. The content of alanine (an inhibitor of pyruvate kinase) reached 2.8 mumol/g in controls and 1.6 mumol/g in hyperammonemic rats, 15 min after ammonium injection. This could explain the different effects of ammonium injection on control and hyperammonemic rats.     

13/14 translocation in a man with reproductive failure

Summary An inherited (maternal origin) 13/14 translocation with the nucleolus organizer regions eliminated and one centromere was found in an oligospermic man whose wife had had two spontaneous abortions. Meiotic studies revealed all stages of spermatogenesis with a trivalent configuration in diakinesis. Possible reasons for his subfertility and his mother's normal fertility are discussed.     

Bacterial periphyton of the Vltava River

Bacterial periphyton formed during 48 hours was studied by glass slide method and direct counting in Vltava River in Praha, Czechoslovakia. At water temperatures 8–11°C the numbers of rods ranged between 24,000 and 336,000 per 1 cm² and those of cocci between 30,000 and 228,000. The relation rods: cocci ranged between 0,9 and 2,4 with an average value of 1,7, whereas in a fishpond this average was 0,5. Among the periphyton 81,3% bacterial cells were active. The rods: cocci relation seems to be a good indicator of water pollution by organic matter, but numbers distinguishing the individual saprobic levels cannot be given yet.

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Zusammenfassung Es wurde der bakterielle Bewuchs von Glassplatten im Vltava-Fluss in Praha untersucht. Zweitägige Exposition erwies sich als am meisten geeignet, um den Bewuchs mikroskopisch durch direkte Zählung quantitativ zu erfassen. In Abhängigkeit von der Tiefe, Wassertemperatur, Verunreinigungsgrad, Sonnenstrahlung u.a. entwickelte sich der Bewuchs quantitativ unterschiedlich und zeigte auch eine unterschiedliche Relation von Stäbchen zu Kokken. Im beta-mesosaproben Bereich wurden im Herbst (Wassertemperaturen 8–11 °C) 24.000 bis 336.000 Stäbchen und 30.000 bis 228.000 Kokken per 1 cm² gefunden, mit den Mittelwerten 163.100 Stäbchen und 104.100 Kokken. Die Vergleichswerte von einem Fischteich in Motol lagen für Stäbchen im gleichen Bereich, während die Kokken, bis 254.000 erreichten.Die Relation Stäbchen: Kokken variierte im Vltava-Fluss zwischen 0,9 (nur einmal) und 2,4 mit dem Mittelwert 1,7, im Fischteich zwischen 0,4 ufnd 0,7 mit dem Mittelwert 0,5. Es hat sich erwiesen, dass diese Relation brauchbare Angaben über die organische Verunreinigung gibt. Leider liegen bisher zu wenige Ergebnisse vor, um eine Skale gegenüber den Saprobitätsstufen zu errichten.Unter den Bewuchsbakterien gab es in der Vltava durchschnittlich 81,3% aktive Zellen, wie durch Fluoreszenzanalyse festgestellt wurde.Fast alle Daten indizieren eine schwächere Verunreinigung des Wassers am linken Ufer als am rechten.

     

Metformin ameliorates olanzapine-induced disturbances in POMC neuron number,axonal projection,and hypothalamic leptin resistance

Antipsychotics have been widely accepted as a treatment of choice for psychiatric illnesses such as schizophrenia. While atypical antipsychotics such as aripiprazole are not associated with obesity and diabetes, olanzapine is still widely used based on the anticipation that it is more effective in treating severe schizophrenia than aripiprazole, despite its metabolic side effects. To address metabolic problems, metformin is widely prescribed. Hypothalamic proopiomelanocortin (POMC) neurons have been identified as the main regulator of metabolism and energy expenditure. Although the relation between POMC neurons and metabolic disorders is well established, little is known about the effects of olanzapine and metformin on hypothalamic POMC neurons. In the present study, we investigated the effect of olanzapine and metformin on the hypothalamic POMC neurons in female mice. Olanzapine administration for 5 days significantly decreased Pomc mRNA expression, POMC neuron numbers, POMC projections, and induced leptin resistance before the onset of obesity. It was also observed that coadministration of metformin with olanzapine not only increased POMC neuron numbers and projections but also improved the leptin response of POMC neurons in the olanzapine-treated female mice. These findings suggest that olanzapine-induced hypothalamic POMC neuron abnormality and leptin resistance, which can be ameliorated by metformin administration, are the possible causes of subsequent hyperphagia.