New mutants resistant to glucose repression affected in the regulation of the NADH reoxidation

Spontaneous mutants resistant to vanadate, arsenate or thiophosphate were isolated from a haploid strain of Saccharomyces cerevisiae. These three anions have an inhibitory effect on some mitochondrial functions and at the level of glyceraldehyde 3-phosphate dehydrogenase, a glycolysis enzyme. All the selected mutants had the same phenotype: they were deficient in alcohol dehydrogenase I, the terminal enzyme of the glycolysis, and possessed a high content of cytochrome c oxidase, the terminal enzyme of the respiratory chain. Moreover, cytochrome c oxidase biosynthesis had become insensitive to the catabolite repression, while the biosynthesis of the other enzymes sensitive to this phenomenon were always inhibited by glucose. Metabolic effects of this pleiotropic mutation manifested themselves in the following ways. 1. Growth rate and final cell mass were enhanced, compared to the wild type, when cells were grown on glucose or on glycerol, but not on lactate or ethanol. 2. Growth under anaerobiosis was nil and mutants did not ferment. 3. Mitochondrial respiration of the mutant strains was identical to the wild type with succinate or 2-oxo-glutarate as substrate, and weak with ethanol. But with added NADH, respiration rate of the mutants was higher than that of the wild type and partially insensitive to antimycin, even when cells were grown in repression conditions. It is postulated that in mutants strains, NADH produced at the level of glyceraldehyde 3-phosphate dehydrogenase, failing to be reoxidized via alcohol dehydrogenase, could be reoxidized with a high turnover owing to the enhancement of the amount of cytochrome c oxidase. Since NADH reoxidation is partially insensitive to antimycin, a secondary pathway going from external NADH dehydrogenase to cytochrome c oxidase is suggested.     

Differential bioavailability of soil-sorbed naphthalene to two bacterial species.

Prediction of the fate of hydrophobic organic contaminants in soils is complicated by the competing processes of sorption and biodegradation. To test the hypothesis that sorbed naphthalene is unavailable to degradative microorganisms, we developed a simple kinetic method to examine the rates and extents of naphthalene degradation in soil-free and soil-containing systems in a comparison of two bacterial species. The method is predicated on the first-order dependence of the initial mineralization rate on the naphthalene concentration when the latter is below the Michaelis-Menten half-saturation constant (Km) for naphthalene for the organism under study. Rates and extents of mineralization were estimated by nonlinear regression analysis of data by using both a simple first-order model and a three-parameter, coupled degradation-desorption model described for the first time here. Bioavailability assays with two bacterial species (Pseudomonas putida ATCC 17484 and a gram-negative soil isolate, designated NP-Alk) gave dramatically different results. For NP-Alk, sorption limited both the rate and extent of naphthalene mineralization, in accordance with values predicted on the basis of the equilibrium aqueous-phase naphthalene concentrations. For strain 17484, both the rates and extents of naphthalene mineralization exceeded the predicted values and resulted in enhanced rates of naphthalene desorption from the soils. We conclude that there are important organism-specific properties which make generalizations regarding the bioavailability of sorbed substrates inappropriate.     

Biochemical studies carried out on different groups ofCandida parapsilosis andCandida rhagii strains by comparing some cellular and mitochondrial activities

A comparative biochemical study was performed on some strains ofCandida rhagii and on strains belonging to different subgroups ofCandida parapsilosis. Measurements of alcohol dehydrogenase activity, resistance to drugs and occurrence of an alternative pathway enabled us to confirm the classification between several subgroups within theC. parapsilosis species.     

Le prétendu rhinocéros de Nouvelle Calédonie est un marsupial (Zygomaturus diahotensis nov. sp.): Solution d'une énigmeet conséquences paléogéographiques

The first mammalian remain ever found in NewCaledonia is an upper tooth found by golddiggers in the Plio-Pleistocene terrace from the Diahot river. This tooth, given to the Muséum national d'Histoire naturelle (Paris) in 1876, was determined as a rhinoceros tooth and then completely forgotten. Its detailed study shows that it belongs to Zygomaturus, a large marsupial diprotodontid genus whose story is rather complicated. The Diahot tooth represents a new species of Zygomaturus, Z. diahotensis nov. sp., close to Z. trilobus from the Australian Pleistocene. That kinship suggests a Plio-Pleistocene land connection between Australia and New Caledonia, whereas till now New Caledonia was supposed to be separated from Australia since the end of the Cretaceous, because of the total absence of indigenous mammals, fossil or recent, in New Caledonia. The latest geological studies in the East Pacific do not contradict our hypothesis.     

A covalently bound photoisomerizable agonist. Comparison with reversibly bound agonists at electrophorus electroplaques

After disulphide bonds are reduced with dithiothreitol, trans-3- (α-bromomethyl)-3’-[α- (trimethylammonium)methyl]azobenzene (trans-QBr) alkylates a sulfhydryl group on receptors. The membrane conductance induced by this “tethered agonist” shares many properties with that induced by reversible agonists. Equilibrium conductance increases as the membrane potential is made more negative; the voltage sensitivity resembles that seen with 50 [mu]M carbachol. Voltage- jump relaxations follow an exponential time-course; the rate constants are about twice as large as those seen with 50 μM carbachol and have the same voltage and temperature sensitivity. With reversible agonists, the rate of channel opening increases with the frequency of agonist-receptor collisions: with tethered trans-Qbr, this rate depends only on intramolecular events. In comparison to the conductance induced by reversible agonists, the QBr-induced conductance is at least 10-fold less sensitive to competitive blockade by tubocurarine and roughly as sensitive to “open-channel blockade” bu QX-222. Light-flash experiments with tethered QBr resemble those with the reversible photoisomerizable agonist, 3,3’,bis-[α-(trimethylammonium)methyl]azobenzene (Bis-Q): the conductance is increased by cis {arrow} trans photoisomerizations and decreased by trans {arrow} cis photoisomerizations. As with Bis-Q, ligh-flash relaxations have the same rate constant as voltage-jump relaxations. Receptors with tethered trans isomer. By comparing the agonist-induced conductance with the cis/tans ratio, we conclude that each channel’s activation is determined by the configuration of a single tethered QBr molecule. The QBr-induced conductance shows slow decreases (time constant, several hundred milliseconds), which can be partially reversed by flashes. The similarities suggest that the same rate-limiting step governs the opening and closing of channels for both reversible and tethered agonists. Therefore, this step is probably not the initial encounter between agonist and receptor molecules.     

Phosphate transport in yeast mitochondria: purification and characterization of a mitoribosomal synthesis dependent proteolipid showing a high affinity for phosphate.

It is possible to obtain from yeast mitochondria a proteolipid able to bind phosphate, by two different procedures. One of them, generally used for lipid extraction, leads to the preparation of a more active crude proteolipid. This crude proteolipid has been purified by various chromatographic procedures and the active fraction, in phosphate binding, is always associated with cardiolipin. Its molecular weight seems to be close to 10000. The phosphate binding shows ligand saturation behavior and is inhibited by arsenate and N-ethylmaleimide; succinate is noninhibitory. This protein seems to be dependent on the mitoribosomal synthesis since it is not present in mitochrondria of mutant "petite colonie" and its amount largely decreases in mitochondria from yeast grown in the presence of chloramphenicol. It is possible to extract a proteolipid from the oligomycin sensitive ATPase, showing the same activity and properties. The hypothesis that this proteolipid acts as a part of the Pi carrier and constitutes the oligomycin-sensitive ATPase complex is discussed.     

Effects of various concentrations of glycerol on post-thaw motility and velocity of human spermatozoa

The percentage post-thaw motility and velocity of semen samples mixed one to one by volume with Ackerman protective medium, with final buffered glycerol concentrations of 6, 8, 12, 16, and 24%, and frozen in liquid nitrogen vapor were studied. Semen frozen in 12 and 16% glycerol gave better motility recovery than those frozen in the other concentrations considered. The changes in motility and kinetics of thawed samples, recorded at 15-min intervals, showed a significant drop 30 min after thawing for motility and after 1 hr for velocity. Results obtained with the photomicrographic method confirm the increase in percentage motility given with the SKM measures when the concentration of glycerol is raised from 8 to 12%. The physical method did not discrimate between progressing and nonprogressing spermatozoa motilities while recording the percentage motility of a population of spermatozoa.     

Lipid peroxidation by peroxidase-catalyzed bioactivation of tyrosine

Tyrosyl free radicals generated by the peroxidase-catalyzed oxidation of peptide tyrosyl residues are known to yield the stable cross-linked product dityrosine. In the present report, horseradish peroxidase is used as a model of peroxidase to study oxidative modifications of non-protein cellular components. Tyrosyl free radicals promote, as many free radicals, the decay of β-phycoerythrin fluorescence emission, they oxidize NADH and ascorbic acid and initiate arachidonic acid peroxidation with formation of hydroperoxides and dienes. These results suggest that tyrosyl free radicals generated when tyrosine residues in protein and peptides are activated in vivo by peroxidase-H₂O₂ might undergo the peroxidation of membrane lipids.