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Biomedical subjects

K Watson

Publications and source records attributed to K Watson.

At least 145 records · Page 8Linked to original sources
PubMed1976

Leucosporidium yeasts: obligate psychrophiles which alter membrane-lipid and cytochrome composition with temperature.

The temperature limits of growth of three psychorphilic yeasts, Leucosporidium frigidum, L. gelidum and L. nivalis, were examined. All species grew well as subzero temperatures (-I degrees C). The maximum temperature of growth was significantly higher with glucose as substrate (18 to 20 degrees C) than with ethanol (17 to 18 degrees C). There was a positive correlation between the growth temperature and the degree of fatty-acid unsaturation of the cell lipids; the lower the temperature, the greater the degree of fatty-acid unsaturation. At subzero temperatures (-I degrees C) with ethanol as substrate, 90% of the total fatty acid was unsaturated with linolenic (35 to 50%) and linoleic (25 to 30%) acids predominating. At temperatures close to the maximum for growth, linolenic acid accounted for less than 20% of the total fatty acid and oleic (20 to 40%) and linoleic (30 to 50%) acids were the major components. Difference spectra of intact cells showed marked changes in the rations and amounts of individual cytochromes as a function of growth temperature. In L.frigidum with glucose as substrate, the ratios of cytochromes a +a3:b:c at 8 and 19 degrees C were I:I-I:2-9 and I:2-3:16-7 respectively. Similar changes in cytochrome ratios were noted for L.gelidum, but changes were much less marked for L. nivalis. The temperature effects were interpreted as supporting the view that membrane structure and composition are fundamental to temperature adaptation in psychrophilic yeasts.

Basidiomycota↗
PubMed1976

Thermal adaptation in yeast: growth temperatures, membrane lipid, and cytochrome composition of psychrophilic, mesophilic, and thermophilic yeasts.

The temperature limits of growth of a number of yeast species were examined, and on this basis the organisms were classified into different thermal categories. The following species were examined: Leucosporidium frigidum and Leucosporidium nivalis, psychrophilic, temperature limits of growth, -2 to 20 degrees C; Canadian lipolytica mesophilic, temperature limits of growth, 5 to 35 degrees Candida parapsilosis and Saccharomyces telluris, thermotolerant, temperature limits of growth, 8 to 42 degrees C; Torulopsis bovina and Candida slooffi, thermophilic, temperature limits of growth, 25 to 45 degrees C and 28 to 45 degrees C, respectively. The membrane lipid and cytochrome composition of mitochrondrial fractions isolated from these yeasts were compared. There was a direct correlation between the growth temperature and the degree of membrane of lipid unsaturation; the lower the temperature, the greater the degree of lipid unsaturation. The membrane lipid composition of the thermophilic yeasts were distinguished by the high percentage (30 to 40%) of saturated fatty acid, as compared with the mesophilic and psychrophilic yeasts. The latter contained approximately 90% unsaturated fatty acid, 55% of which was linolenic acid, C alpha-18:3. Changes in phospholipid composition in relation to temperature were also noted. The respiratory-deficient thermophile, C. slooffi, was characterized by the absence of cardiolipin (sensitivity 0.1 mug of phosphorus) and cytochrome aa3. The absence of conventional mitochondrial structures in this thermophilic microorganism is tentatively suggested although low concentrations of cytochromes b, c, and c1 were detected by low-temperature spectroscopy. On the other hand, the respiratory-competent thermophile, T. bovina, was characterized by a high cardiolipin (25% of the total phospholipid) and cytochrome aa3 content (1 nmol/mg of mitochrondrial protein). Low-temperature spectra showed the presence of one b-type cytochrome in the thermophilic yeasts, two b-type cytochromes in the mesophilic yeasts, and three b-type cytochromes in the psychrophilic yeasts. It was concluded that a knowledge of the properties of the biological membrane is fundamental to an understanding of the ability of a microorganism to grow and reproduce in different temperature environments.

Adaptation, Physiological↗
PubMed1976

Isolation and identification of paracetamol metabolites.

A comparison has been made of the urinary metabolites of volunteers who had taken therapeutic doses of paracetamol with those of persons who had taken an overdose in an attempt to highlight the metabolic changes associated with massive doses. The main technique for examining urine samples was two-dimensional thin layer chromatography. Other chromatographic techniques were used for the isolation and purification of metabolites. The urinary metabolites after a therapeutic dose of paracetamol were identified as free paracetamol, paracetamol sulphate, 3-hydroxy-paracetamol-3-sulphate, 3-methoxy-paracetamol sulphate, paracetamol glucuronide, 3-methoxy-paracetamol glucuronide, paracetamol 3-cysteine conjugate and paracetamol 3-mercapturate. The same metabolites were also present in urine following overdosage but the proportions were quite different. There was particularly a big increase in the relative amounts of cysteine and mercapturic acid conjugates excreted. No new metabolites were found. The significance of these findings is briefly discussed in relation to the metabolism and toxicology of paracetamol.

Acetaminophen↗
PubMed1975

Phase transitions in yeast mitochondrial membranes. The effect of temperature on the energies of activation of the respiratory enzymes of Saccharomyces cerevisiae.

The effect of temperature on the activation energies of mitochondrial enzymes of the yeast Saccharomyces cerevisiae was examined. Non-linear Arrhenius plots with discontinuities in the temperature range 14-19 degrees C and 19-22 degrees C were observed for the respiratory enzymes and mitochondrial ATPase (adenosine triphosphatase) respectively. A straight-line Arrhenius plot was observed for the matrix enzyme, malate dehydrogenase. The activation energies of the enzymes associated with succinate oxidation, namely, succinate oxidase, succinate dehydrogenase and succinate-cytochrome c oxidoreductase, were in the range 60-85kJ/mol above the transition temperature and 90-160kJ/mol below the transition temperature. In contrast, the corresponding enzymes associated with NADH oxidation showed significantly lower activation energies, 20-35kJ/mol above and 40-85kJ/mol below the transition temperature. The discontinuities in the Arrhenius plots were still observed after sonication, treatment with non-ionic detergents or freezing and thawing of the mitochondrial membranes. Discontinuities for cytochrome c oxidase activity were only observed in freshly isolated mitochondria, and no distinct breaks were observed after storage at -20 degrees C. Mitochondrial ATPase activity still showed discontinuities after sonication and freezing and thawing, but a linear plot was observed after treatment with non-ionic detergents. The results indicate that the various enzymes of the respiratory chain are located in a similar lipid macroenvironment within the mitochondrial membrane.

Adenosine Triphosphatases↗
PubMed1975

Membrane-lipid unsaturation and mitochondrial function in Saacharomyces cerevisiae.

The lipid composition of yeast cells was manipulated by the use of an unsaturated fatty acid auxotroph of Saccharomyces cerevisiae. There was a 2-3-fold decrease in the concentration of cytochromes a+a3 when the unsaturated fatty acid content of the cells was decreased from 60-70% of the total fatty acid to 20-30%. The amounts of cytochromes b and c were also decreased under these conditions, but to a lesser extent. Further lipid depletion, to proportions of less than 20% unsaturated fatty acid, led to a dramatic decrease in the content of all cytochromes, particularly cytochromes a+a3. The ATPase (adenosine triphosphatase), succinate oxidase and NADH oxidase activities of the isolated mitochondria also varied with the degree of unsaturation of the membrane lipids. The lower the percentage of unsaturated fatty acid, the lower was the enzymic activity. Inhibition of mitochondrial ATPase by oligomycin, on the other hand, was not markedly influenced by the membrane-lipid unsaturation. Npn-linear Arrenius plots of mitochondrial membrane-bound enzymes showed transition temperatures that were dependent on the degree of membrane-lipid unsaturation. The greater the degree of lipid unsaturation, the lower was the transition temperature. It was concluded that the degree of unsaturation of the membrane lipids plays an important role in determining the properties of mitochondrial membrane-bound enzymes.

Adenosine Triphosphatases↗
PubMed1970

Biogenesis of mitochondria. 13. The isolation of mitochondrial structures from anaerobically grown Saccharomyces cerevisiae.

Morphologically intact structures have been isolated from anaerobically grown yeast cells which have many of the properties of yeast mitochondria. The structures are about 0.5 micro in diameter and contain malate dehydrogenase, succinate dehydrogenase, oligomycin-sensitive ATPase, and DNA of buoyant density 1.683 g/cc, characteristic of yeast mitochondria. The morphology of the structures is critically dependent on their lipid composition. When isolated from cells grown anaerobically in the presence of supplements of unsaturated fatty acid and ergosterol, their unsaturated fatty acid content is similar to that of mitochondria from aerobically grown cells. These lipid-complete structures consist pre-dominantly of double-membrane vesicles enclosing a dense matrix which contains a folded inner membrane system bordering electron-transparent regions which are somewhat different from the cristae of functional mitochondria. In contrast, the structures from cells grown without lipid supplements are much simpler in morphology; they have a dense granular matrix surrounded by a double membrane but have no obvious folded inner membrane system within the matrix. The lipid-depleted structures are very fragile and are only isolated in intact form from protoplasts that have been prefixed with glutaraldehyde

Adenosine Triphosphatases↗
PubMed1968

Respiratory components of Aspergillus niger mitochondria.

The respiratory components of tightly coupled mitochondria from the filamentous fungus Aspergillus niger were studied. Cytochromes a + a(3), b, and c + c(1) were identified by difference spectra. The cytochrome spectra were qualitatively similar to yeast and rat liver mitochondria. The mitochondria contained, per gram of protein, an average of 2.9 and 7.0 mumoles of ubiquinone and nicotinamide adenine dinucleotide, respectively.

Aspergillus↗