Nucleotide pools of growing,synchronized and stressed cultures of Saccharomyces cerevisiae

High pressure liquid chromatography has been used to study the acid soluble nucleotide pool of Saccharomyces cerevisiae under different conditions of growth. ATP, ADP, AMP, NAD, GTP, UTP, UDP, CTP, CDP, and UDP-sugars plus UMP could be separated and were found in concentrations higher than 0.1 mumol per g yeast cell dry weight (= detection limit). During glucose-limited continuous culture the levels of individual nucleotides depended on the growth rate, which was most pronounced with pyrimidine (uridine, cytidine) nucleotides. The energy charge (E.C.) remained high (0.9) at all growth rates (0.07-0.3 h-1). During synchronized growth at a constant growth rate (0.11 h-1) almost all nucleotide levels and the E.C. remained at constant values with the only exception of UDP-sugars and UMP of which increased levels were found during the phase of budding. Under conditions of metabolic stress (addition of antimycin A, deoxyglucose plus iodoacetate) pronounced changes in the levels of purine (adenine and guanine) nucleotides and the E.C. were observed. All other nucleotides were less influenced by these conditions. Only under these conditions IMP accumulation was observed. The results strongly argue against the significance of purine nucleotide or E.C. measurements under viable conditions. In contrast, changes in the levels of pyrimidine nucleotides seem to be indicative of changes in the flux through the metabolic pathways where they act as coenzymes.     

Effects of potassium,lanthanum and black widow spider venom on miniature synaptic potentials in theTorpedo electroplax

Summary Using a method of focal drug application it is demonstrated that high potassium concentration, lanthanum, and black widow spider venom accelerate spontaneous transmitter release inTorpedo electric tissue.This investigation was supported by the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 138). Thanks are due to Dr. R. Martin and the staff of the Stazione Zoologica, Naples, for supplyingTorpedo, and Dr. N. Frontali, Rome, for a gift of frozen black widow spiders.     

The effect of energization on the apparent Michaelis–Menten constant for oxyge in nmitochondrial respiration

Lineweaver-Burk plots of 1/v against 1/[O(2)] for rat liver mitochondrial respiration with succinate or ascorbate+NNN'N'-tetramethyl-p-phenylenediamine as substrates are non-linear. In state 3u (uncoupled by trifluoromethoxycarbonyl cyanide phenylhydrazone) such plots tend to be concave upward, whereas in state 4 (energized) the plots were concave downward. The apparent K(m) for oxygen is larger in state 4 than in state 3u, despite the higher turnover in the latter system. It is postulated that at least one reversible reaction occurs between cytochrome c and cytochrome c oxidase, whose rate is increased on energization (reversed electron transfer); a model including such a reaction is proposed which accounts semiquantitatively for the observations.     

Effect of arterial perfusion on net water flux and active sodium transport across the isolated skin ofBufo bufo bufo (L.)

Summary A method has been developed which allows perfusion of the blood vessels in isolated pelvic skins ofB. bufo. The effect of various doses of vasotocin (AVT) on net water flux (inside medium 220 mOsM, outside medium 11 mOsM) and active sodium transport were compared in perfused and unperfused skins. The unstimulated water flux (Fig. 3) and the active sodium transport (Fig. 6) were unaffected by perfusion. The threshold for stimulation of water flux was between 0.01 and 0.1 nM vasotocin in perfused skins and between 0.1 and 1 nM in unperfused skins. The threshold for stimulation of active sodium transport was between 0.005 and 0.05 nM vasotocin in perfused skins and between 0.05 and 0.5 nM in unperfused skins. The maximal water flux through perfused skins, 4 l/cm²min, was obtained at 1 nM vasotocin. At 10 nM vasotocin the water flux was only 0.7 l/cm²min in unperfused skins. The maximal active sodium transport was approximately of the same magnitude in perfused and unperfused skins, at 0.5 mM and at 50 nM, respectively.     

Deformation of the Diastolic Left Ventricle: I. Nonlinear Elastic Effects

A linear incremental finite element model is used to analyze the mechanical behavior of the left ventricle. The ventricle is treated as a heterogeneous, non-linearly elastic, isotropic, thick-walled solid of revolution. A new triaxial constitutive relation for the myocardium is presented which exhibits the observed exponential length-passive tension behavior of left ventricular papillary muscle in the limit of uniaxial tension. This triaxial relation contains three parameters: (a) a “small strain” Young's modulus, (b) a Poisson's ratio, and (c) a parameter which characterizes the nonlinear aspect of the elastic behavior of heart muscle. The inner third and outer two-thirds of the ventricular wall are assumed to have small strain Young's moduli of 30 and 60 g/cm², respectively. The Poisson's ratio is assumed to be equal to 0.49 throughout the ventricular wall. In general, the results of this study indicate that while a linearly elastic model for the ventricle may be adequate in terms of predicting pressure-volume relationships, a linear model may have serious limitations with regard to predicting fiber elongation within the ventricular wall. For example, volumes and midwall equatorial circumferential strains predicted by the linear and nonlinear models considered in this study differ by approximately 20 and 90%, respectively, at a transmural pressure of 12 cm H₂O.     

Synthese der Hämolymphproteine und die Aufnahme der Dotterfraktion in die Oocyte unter Actinomycin-Einflu\ (Untersuchungen anMusca domestica)

Zusammenfassung Mit Hilfe von autoradiographischen und elektrophoretischen Methoden wurde die Dottereinlagerung in den wachsenden Oocyten vonMusca domestica untersucht. Sie beginnt nach 30 min im Autoradiogramm sichtbar zu werden. Durch ihre Färbbarkeit und Markierung konnte die Dotterfraktion im Pherogramm von Ovar und Hämolymphe eines mittleren Wachstumsstadiums (Stadium 3) nachgewiesen werden. Nach Abschlu\ der Vitellogenese tritt sie in der Hämolymphe nicht mehr auf. Die Einlagerung der Dotterproteine wird durch Actinomycin gestört, dagegen läuft ihre Synthese nahezu unbeeinflu\t weiter. Die Transporthemmung kann als bisher unbekannter Nebeneffekt des Actinomycins gedeutet werden.

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Synthesis of haemolymph proteine and the uptake of the yolk fraction in the oocyte during Actinomycin-treatment. (Studies onMusca domestica)

Summary By means of radioautographic and electrophoretic techniques yolk protein uptake in the growing oocytes ofMusca domestica was investigated. After 30 min yolk protein becomes visible in the radioautograms. By stainability and labeling the yolk fraction could be demonstrated in the pherogram of ovary and haemolymph in an intermediate developmental stage (stage 3). After the end of vitellogenesis it does not appear in the haemolymph. The yolk protein uptake is inhibited by Actinomycin, but the synthesis goes on nearly as normal. This inhibition can be interpretated as a new accessory effect of Actinomycin.