Cells from stationary-phase cultures of two strains of Saccharomyces cerevisiae (3 and 20) failed to flocculate when grown in a complex or a chemically defined medium, while those of two other strains (11 and 13) flocculated when grown in either medium. Strain 30 flocculated when grown in complex but not defined medium and harvested from stationary-phase cultures. pH-electrophoretic mobility measurements on all five strains showed that mobility attributable to carboxyl groups usually increased as cultures progressed from the exponential to the stationary phase, while that caused by phosphate groups tended to decline. Acquisition of flocculating ability was accompanied in strains 11 and 30 by a slight increase in amidase activity, and greater increases compared with nonflocculent populations in activities of leucine aminopeptidase. alpha-mannosidase, and proteinase C. Activities of proteinases A and B showed no correlation with acquisition of flocculating ability.
Saccharomyces cerevisiae, grown aerobically or anaerobically under conditions which induce a requirement for a sterol and an unsaturated fatty acid, synthesized approximately the same amounts of neutral lipid and intracellular low-density vesicles, although the neutral lipids in aerobically-grown cells contained more esterified sterol and less triacylglycerol than those in anaerobically-grown cells. Kluyveromyces fragilis synthesized much less neutral lipid and a smaller quantity of low-density vesicles than S. cerevisiae whether grown at 30 degrees C (generation time 1.1 h) or 20 degrees C (generation time 2.1 h). Both yeasts synthesized highly saturated triacylglycerols, relatively unsaturated phospholipids, and esterified sterols with an intermediate degree of unsaturation irrespective of the conditions under which they were grown. Free sterols in the yeasts were rich in ergosterol and 22(24)-dehydroergosterol, while the esterified sterol fractions were richer in zymosterol.
1. A simple in vitro technique that predicts drug transfer into breast milk is described. 2. Drugs of differing physical and chemical characteristics were tested. 3. The technique provides an experimental system for studying plasma to milk transfer with changing milk composition. 4. A mechanism proposing a role of milk proteins in controlling drug entry into milk is discussed.
A mixture of small (0.43-Am diameter) and large (0.62-,um diameter) lowdensity vesicles from spheroplasts of Saccharomyces cereuisiae was fractionated by rate centrifugation in a gradient of 0 to 8% (wt/vol) Ficoll to yield fractions rich (90 to 95%) in small or large vesicles. The large, but not small, vesicles swelled when diluted into mannitol solutions containing less than 0.4 M mannitol. The pH-electrophoretic mobility curve of the large vesicles showed that they are probably enclosed in a phospholipid-protein membrane. The dyes neutral red and toluidine blue, accumulated into large vesicles by intact cells and spheroplasts, were largely lost from large vesicles when these were separated from stained spheroplasts. Sudan black III stained small and large vesicles, both classes of vesicle retaining the stain on separation. Fractions rich in large vesicles contained proportionately more phospholipid and less free sterols, diacylglycerols, and free fatty acids compared with those enriched in small vesicles. The two classes of vesicles contained about the same proportions of esterified sterols and triacylglycerols. The free fatty acids in both small and large vesicles were free from unsaturated fatty-acyl residues; diacylglycerols and triacylglycerols contained appreciable proportions of unsaturated fatty-acyl residues. Small vesicles were richer in lipase activity, whereas the larger vesicles contained greater f3-glucanase and a-mannosidase activities. Phospholipase activity could not be detected in any of the fractions.
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