Genetic aspects of ethanol tolerance and production bySaccharomyces cerevisiae

We developed a method of hybrid selection between homothallic wild-type and heterothallic strains. The hybrids obtained were used to study the heredity of ethanol tolerance and production. Both characters segregated independently, but no ethanol-sensitive strains were able to produce high levels of ethanol. At least four genes are implicated in ethanol tolerance.     

Selection of Wine Yeasts for Growth and Fermentation in the Presence of Ethanol and Sucrose

To optimize the conversion of carbohydrates to ethanol, strains of several Saccharomyces species were examined for the ability to grow and ferment in a range of sucrose and ethanol concentrations. A total of 632 wine yeasts, most of them isolated from wineries in Andalusia and Extremadura, southwestern Spain, were subjected to screening and selection. Growth and fermentative capacity in different ethanol and sucrose concentrations varied from one strain to another. There was no correlation between growth and fermentative capacity. The best 35 strains grew in 15% ethanol and fermented in 18% ethanol. Ethanol accumulated, although at a reduced rate, after the cells stopped growing. Most yeast strains were highly fermentative in 50% sucrose. Some of them effectively utilized the carbohydrates of the culture, yielding final ethanol concentrations of > 14%. Of the 35 selected strains, 16 were promising for genetic analysis and breeding because of their capacity to sporulate. These strains were homothallic, and their spores were viable. The meiotic products analyzed so far were also homothallic.     

Generation of an internal matrix in mature avian erythrocyte nuclei during reactivation in cytoplasts

When fused with mouse L-cell cytoplasts, chick erythrocyte nuclei enlarge, take up proteins from the host cytoplasm, and recommence RNA synthesis. We found that during this transition the erythrocyte nuclei gain an internal nuclear matrix, thus providing a novel approach to questions concerning the nature of the salt-resistant intranuclear skeleton. A new method for preparation and examination of the nuclear matrix in situ is also described.     

Study of Chromosomes in the Newborn after Ultrasonic Fetal Heart Monitoring in Labour

The effect of continuous-wave ultrasound on the chromosomes of newborn infants has been investigated. Twenty-four women were studied during labour. The fetal heart was monitored by a Sonicaid FM2 monitor applied to the abdomen, and continuous monitoring undertaken for intervals varying from 1 hour 5 minutes to 9 hours 25 minutes. There was no increase in the number of chromosome aberrations in cultures of blood taken from the insonated babies when compared with controls.     

Adrenomedullary Secretion of DOPA, Catecholamines, Catechol Metabolites, and Neuropeptides

Abstract: Catecholamines and their metabolites have been proposed as markers of sympathetic nervous system stimulation. However, the adrenal medulla is a rich source of catecholamines and catecholamine metabolites and may play a significant role in plasma levels of these compounds. In addition to adrenal catecholamine metabolite efflux, the role of the catecholamine precursor 3,4-dihydroxyphenylalanine (DOPA) has not been fully evaluated. The simultaneous effluxes of catecholamines, metabolites, DOPA, and neuropeptides were measured in perfusates from isolated dog adrenals. The relative abundance of compounds detected consistently during unstimulated conditions was epinephrine ≫ norepinephrine > 3,4-dihydroxyphenylglycol > metanephrine > normetanephrine > dopamine > 3,4-dihydroxyphenylacetic acid > 3-methoxy-4-hydroxyphenylglycol ≥ DOPA ≫ [Met]enkephalin ≫ neuropeptide Y. Effluxes of analytes were not affected by cocaine and the ratios of catecholamines to metabolites increased dramatically with carbachol stimulation, consistent with negligible reuptake into adrenal cells. Thus, most of the 3,4-dihydroxyphenylglycol is expected to be derived from epinephrine and norepinephrine subsequent to translocation from chromaffin vesicles into the cytosol. The efflux of DOPA increased dramatically during stimulation with 30 µ M carbachol in a calcium-dependent manner. Efflux of DOPA during the initial stabilization period of the perfusion preparation declined exponentially, in parallel with the effluxes of the catecholamines and neuropeptides but not with metabolites. Evoked release of DOPA was Ca²⁺-dependent. These data suggest that DOPA can be stored and released exocytotically from chromaffin granules.     

Production and standing stocks of the kelp Macrocystis laevis Hay at the Prince Edward Islands,Subantarctic

Summary The recently described species Macrocystis laevis Hay is endemic to the Prince Edward Islands. Aerial photographs of Marion Island were used to outline the distribution of the kelp and to assess its cover. M. laevis occurs along the lee shore of the island, between the 5 and 20 m isobaths. Plant densities and gross plant morphology were measured by divers during April/May 1988. Net production was estimated from growth measurements taken in April/May 1988 and 1989 and again during August 1989. The mean biomass of kelp was 0.67 kgC·m^–2 within the kelp beds. Net production was estimated at 7.7 gC·m^–2·d^–1 and 11.5 gC·m^–2d^–1 during the months of April and August respectively. M. laevis had a uniform frond-length frequency distribution, which suggests that only the oldest fronds are lost by wave action or senescence. Based on calculations for M. laevis and Durvillaea antarctica (the two species making up most of the macrophyte biomass) macrophytes are more productive per unit area than the phytoplankton but contribute less to the seas around the Prince Edward Islands by virtue of their small spatial coverage. Neither of the kelps lose much material as particulate or dissolved organic carbon through fragmentation. The extent of grazing on live M. laevis fronds is unknown, and only D. antarctica contributes to a macrofaunal detrital community. The contribution of M. laevis production to the nearshore ecology of the islands seems limited, as we suspect that almost all of its production is exported to the open ocean pelagic system.     

Plasmalemma Chloride Transport in Chara corallina: Inhibition by 4,4'-Diisothiocyano-2,2'-Disulfonic Acid Stilbene

Chloride transport, presumably via a Cl⁻-2H⁺ co-transport system, was investigated in Chara corallina. At pH 6.5, the control influx (3.1 picomoles per centimeter² per second) was stimulated 4-fold by an 18-hour Cl⁻ starvation. The stimulated influx was inhibited to 4.7 picomoles per centimeter² per second after a 60-minute pre-exposure to 0.5 millimolar 4,4′-diisothiocyano-2,2′-disulfonic acid stilbene (DIDS). This compares with a nonsignificant inhibition of the control under similar conditions. At 2 millimolar DIDS, both stimulated and control influx were inhibited to values of 1.1 and 2.2 picomoles per centimeter² per second, respectively; in all cases, DIDS inhibition was reversible. Over the pH range 4.8 to 8.5, the control and DIDS-inhibited influx showed only slight pH sensitivity; in contrast, the stimulated flux was strongly pH dependent (pH 6.5 optimum). Inasmuch as changes in pH alter membrane potential, N-ethylmaleimide was used to depolarize the membrane; this had no effect on Cl⁻ influx. A transient depolarization of the membrane (about 20 millivolts) was observed on restoration of Cl⁻ to starved cells. The membrane also depolarized transiently when starved cells were exposed to 0.5 millimolar DIDS, but the depolarization associated with Cl⁻ restoration was inhibited by a 40-minute pretreatment with DIDS. Exposure of control cells to DIDS caused only a small hyperpolarization (about 7 millivolts). DIDS may have blocked Cl⁻ influx by inhibiting the putative plasmalemma H⁺-translocating ATPase. Histochemical studies on intact cells revealed no observable effect of DIDS on plasmalemma ATPase activity. However, DIDS application after fixation resulted in complete inhibition of ATPase activity.

The differential sensitivity of the stimulated and control flux to inhibition by DIDS may reflect an alteration of transport upon stimulation, but could also result from differences in pretreatment. The stimulated cells were pretreated with DIDS in the absence of Cl⁻, in contrast to the presence of Cl⁻ during pretreatment of controls. The differential effect could result from competition between Cl⁻ and DIDS for a common binding site. Our histochemical ATPase results indicate that Cl⁻ transport and membrane ATPase are separate systems, and the latter is only inhibited by DIDS from the inside of the cell.

     

Measles virus inhibits acquisition of lymphocyte functions but not established effector functions

The effect of measles-virus infection on effector activities of human lymphocytes and on the generation of certain effector activities was studied in vitro. Addition of measles virus to allogeneic mixed lymphocyte cultures resulted in a strongly depressed cytolytic activity in a subsequent cell-mediated lympholysis assay. Late addition of measles virus did not inhibit cytotoxic effector function, although effector cells were probably infected. Similarly, measles-virus infection did not affect the ability of lymphocytes to mediate antibody-dependent cellular cytotoxicity. Addition of measles virus to lymphocytes with, or shortly after, exposure of the cells to the polyclonal activator pokeweed mitogen resulted in abolition of the synthesis of immunoglobulins in vitro. When the virus was added late, the rate of Ig secretion was only partially inhibited. Finally, when lymphocytes were cultured without stimulus in medium supplemented with fetal bovine serum, a population of inhibitory cells was generated. Measles virus was able to prevent the generation of such inhibitory cells. In conclusion, measles virus inhibited acquisition of various effector functions, but the activities of committed lymphocytes were generally not affected.     

Ectogalactosyltransferase studies in fibroblasts and concanavalin A- stimulated lymphocytes

In this communication, we have demonstrated that hydrolysis of the nucleotide sugar can cause errors in the detection of an ectoglycosyltransferase. Spleen cell suspensions can incorporate radioactivity when incubated with labeled UDP-galactose, but all the activity is due to decomposition of the nucleotide sugar and uptake of the free sugar. The fibroblast cell lines can incroporate carbohydrate directly from UDP-galactose. Several criteria are presented with can be used to demonstrate that a nucleotide sugar is the direct carbohydrate donor.