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PubMed1994

Ethanol potentiates the clastogenicity of pan masala--an in vitro experience.

The significance of the interaction between alcohol and tobacco in causing head and neck cancers is well documented. Our previous reports on in vitro studies using aqueous and organic extracts as well as cytogenetic studies among pan masala consumers have conclusively shown the genotoxic potential of pan masala--a dry mixture of the areca nut, lime, catechu, unspecified flavouring agents, etc., often containing tobacco in it and is widely consumed in India. Now in the present report, the clastogenic effect of ethanol and pan masala in different combinations was evaluated on Chinese hamster ovary cells utilizing chromosome aberration (CA) frequency as an endpoint. An ethanol concentration of up to 2.0% had no effect on CA/cell value. The low-dose continuous treatment and high-dose short-term pre-, post- and simultaneous treatment of ethanol and aqueous extract of pan masala with and without tobacco yielded dose-dependent elevations in CA frequency, compared to any of these two substances alone. Thus, these results provide evidence that alcohol consumption may potentially increase the risk of oral cancer among pan masala chewers.

Animals↗
PubMed2001

The effect of ethanol and acetate on protein expression in Pichia pastoris.

Pichia pastoris is an excellent host for high-level heterologous gene expression, but there is still much interest in improving the productivity of recombinant protein production. P. pastoris produces a small amount of ethanol as a by-product during the glycerol fed-batch phase and the mixed-feed induction phase (glycerol-methanol) of high cell density fermentations, regardless of the phenotype (Mut+, Mut(s), or Mut-). We have nvestigated ethanol repression of the AOX1 promoter using strains, GS115 (Mut+) and MC100-3 (Mut-), expressing an AOX1-lacZ fusion. The addition of 10 mg l(-1) ethanol at the start of methanol induction delayed beta-galactosidase production and methanol utilization for four hours in shake flask experiments. When ethanol and acetate were added together, all of the ethanol was converted to acetate, which also represses the AOX1 promoter. The effects of ethanol and acetate on protein expression in P. pastoris at shake flask and fermentor conditions are discussed.

Journal Article↗
PubMed2003

Evolutionary engineering of Saccharomyces cerevisiae for anaerobic growth on xylose.

Xylose utilization is of commercial interest for efficient conversion of abundant plant material to ethanol. Perhaps the most important ethanol-producing organism, Saccharomyces cerevisiae, however, is incapable of xylose utilization. While S. cerevisiae strains have been metabolically engineered to utilize xylose, none of the recombinant strains or any other naturally occurring yeast has been able to grow anaerobically on xylose. Starting with the recombinant S. cerevisiae strain TMB3001 that overexpresses the xylose utilization pathway from Pichia stipitis, in this study we developed a selection procedure for the evolution of strains that are capable of anaerobic growth on xylose alone. Selection was successful only when organisms were first selected for efficient aerobic growth on xylose alone and then slowly adapted to microaerobic conditions and finally anaerobic conditions, which indicated that multiple mutations were necessary. After a total of 460 generations or 266 days of selection, the culture reproduced stably under anaerobic conditions on xylose and consisted primarily of two subpopulations with distinct phenotypes. Clones in the larger subpopulation grew anaerobically on xylose and utilized both xylose and glucose simultaneously in batch culture, but they exhibited impaired growth on glucose. Surprisingly, clones in the smaller subpopulation were incapable of anaerobic growth on xylose. However, as a consequence of their improved xylose catabolism, these clones produced up to 19% more ethanol than the parental TMB3001 strain produced under process-like conditions from a mixture of glucose and xylose.

Aldehyde Reductase↗
PubMed1984

Interaction between ethanol and low dietary zinc during gestation and lactation in the rat.

Pregnant rats were fed individual liquid diets throughout gestation and lactation. The diets contained either 2 or 10 micrograms zinc/ml diet with or without 30% of the kilocalories supplied from ethanol. The low zinc diet produced a moderate zinc deficiency in dams evidenced by decreases in tissue zinc content, serum alkaline phosphatase activity and urinary zinc concentration. Despite the presence of high zinc content in the diet, ethanol antagonized the maternal zinc status to a level typical of that produced by the low zinc diet. The lowest zinc status, however, was found when low dietary zinc and ethanol were combined. The maternal interaction between ethanol and zinc also depressed offspring serum zinc and alkaline phosphatase activity in a similar manner. Ethanol, however, did not affect tissue content of calcium, magnesium or phosphorus, which indicates that ethanol is a specific antagonist of zinc utilization during gestation and lactation.

Alkaline Phosphatase↗
PubMed2006

Application of heated inlet extensions to the TSI 3306/3321 system: comparison with the Andersen cascade impactor and next generation impactor.

Pharmaceutical aerosol size distribution analysis based on multi-stage inertial impaction is well accepted, though laborious. The TSI 3306 Impactor Inlet/3321 time-of-flight (TOF) Aerodynamic Particle Size Analyzer (APS) has been evaluated for its ease of use and potential for time savings during product development. However, instrument inlet modifications may be necessary for increased correlation with equivalent measurements obtained by inertial impaction following pharmacopeial methods. A heated inlet extension tube was located between the USP/Ph.Eur. throat and the Single-Stage Impactor (SSI) to promote evaporation of residual ethanol from aerosol droplets, generated from two formulations containing ethanol as semi-volatile solubilizer (8 and 20% w/w) for the active pharmaceutical ingredient. As temperature and extension length increased, the SSI-measured fine particle fraction (aerosol < 4.7 microm aerodynamic diameter) also increased, for the aerosols used in this study. These values correlated quite closely with equivalent measures made by multi-stage cascade impactor equipped with the same throat. Particle size distribution profiles measured with the APS for either formulation did not significantly change utilizing the heated extensions, suggesting that ethanol evaporation was largely complete at any condition by the time the aerosol entered the measurement zone of the TOF analyzer. The addition of a heated inlet extension may be useful to facilitate evaporation of residual semi-volatile species, especially when an agreement of APS-derived particle size mass distribution data from the SSI with multi-stage cascade impactors is desired. However, complete evaporation of the semi-volatile species may not be necessary for SSI-generated mass distribution to match conventionally used cascade impactors.

Administration, Inhalation↗
PubMed1991

Suppression of immune responsiveness: sex differences in prenatal ethanol effects.

Exposure to ethanol in utero results in changes in the offspring's developing immune system, including thymus lymphocyte subpopulation shifts and functional lymphocyte changes that persist in adult animals. The present study was designed to define further the extent of changes in the immune system that result from fetal ethanol exposure and to compare effects in male and female offspring. In adulthood, male and female offspring from Sprague-Dawley dams fed an ethanol-containing liquid diet (alcohol, A), an isocaloric liquid control diet (pair-fed, PF), or laboratory chow and water (control, C) during pregnancy were tested for several measures of immune competency. Prenatal ethanol exposure differentially affected male and female offspring. Fetal ethanol-exposed males exhibited a decrease in thymocyte number as well as a decreased splenic lymphocyte proliferative response to the T-cell mitogen, concanavalin A (Con A), with a concomitant decrease in recoverable blast cells, when compared with PF and C males. Further, the defect in T-cell proliferation of A males was not due to an inability to produce the critical growth factor, interleukin-2 (IL-2), but to an inability of lymphoblasts to utilize exogenous IL-2. Fetal ethanol-exposed females showed some suggestion of lower thymocyte counts and decreased splenic T-cell proliferative responses to Con A compared to PF and C females. For most of the immune measures, however, no significant differences occurred among A, PF, and C females. In utero ethanol exposure did not significantly alter spleen cell counts or IL-2 production, splenic B-cell proliferation to bacterial lipopolysaccharide (LPS), or thymocyte response to IL-2 in animals of either sex.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1982

Antimycin inhibition as a probe of mitochondrial function in isolated rat hepatocytes. Effects of chronic ethanol consumption.

Previous studies have established that hepatic mitochondria and submitochondrial particles from rats, fed ethanol chronically, display diminished respiratory activities and alterations in the contents of specific electron transfer chain components. The latter include a decrease of about 50% in cytochrome b content. Titrations of respiratory activity in submitochondrial particles with antimycin, a stoichiometric inhibitor of electron flow through the cytochrome b-c1 region of the respiratory chain, indicated a comparable decrease (35%) in the amount of antimycin required to elicit maximal inhibition ('titer') after chronic ethanol treatment. Measurements of antimycin binding to submitochondrial particles by fluorescence quenching demonstrated a similar diminution in the number of tight binding sites per mg protein. By contrast, hepatocytes isolated from control and ethanol-fed rats exhibited nearly identical rates of oxygen utilization under a variety of conditions. However, antimycin titrations of respiratory activity in isolated hepatocytes revealed a 60% decrease in the antimycin titer, but no change in the maximal extent of inhibition after chronic ethanol treatment. Direct measurements of cytochrome b which could be reduced in the presence of antimycin in hepatocytes confirmed a comparable decrease (42%) after chronic ethanol treatment. The results demonstrate that molecular alterations in the cytochrome b region of the respiratory chain caused by ethanol feeding are present in intact liver cells, but suggest that substrate accessibility, rather than the respiratory chain, limits the rate of oxygen utilization in isolated hepatocytes. The data also suggest that mitochondria account for at least 80% of total oxygen utilization by liver cells from both control and ethanol-fed rats.

Alcoholism↗
PubMed2004

Fermentation of biomass-generated producer gas to ethanol.

The development of low-cost, sustainable, and renewable energy sources has been a major focus since the 1970s. Fuel-grade ethanol is one energy source that has great potential for being generated from biomass. The demonstration of the fermentation of biomass-generated producer gas to ethanol is the major focus of this article in addition to assessing the effects of producer gas on the fermentation process. In this work, producer gas (primarily CO, CO(2), CH(4), H(2), and N(2)) was generated from switchgrass via gasification. The fluidized-bed gasifier generated gas with a composition of 56.8% N(2), 14.7% CO, 16.5% CO(2), 4.4% H(2), and 4.2% CH(4). The producer gas was utilized in a 4-L bioreactor to generate ethanol and other products via fermentation using a novel clostridial bacterium. The effects of biomass-generated producer gas on cell concentration, hydrogen uptake, and acid/alcohol production are shown in comparison with "clean" bottled gases of similar compositions for CO, CO(2), and H(2). The successful implementation of generating producer gas from biomass and then fermenting the producer gas to ethanol was demonstrated. Several key findings following the introduction of producer gas included: (1) the cells stopped growing but were still viable, (2) ethanol was primarily produced once the cells stopped growing (ethanol is nongrowth associated), (3) H(2) utilization stopped, and (4) cells began growing again if "clean" bottled gases were introduced following exposure to the producer gas.

Bioreactors↗
PubMed1983

The interaction of ethanol and exogenous arachidonic acid in the formation of leukotrienes and prostaglandin D2 in mastocytoma cells.

The present studies were undertaken to examine the hypothesis that ethanol could affect cellular biosynthesis in the murine mastocytoma cell of prostaglandins and leukotrienes, oxidative metabolites of arachidonic acid, at concentrations that could be encountered in vivo as well as during in vitro experiments. The effects of ethanol which encompass these concentration ranges (200-1000 mg%) can be summarized as follows: first in the absence of exogenous arachidonic acid, ethanol caused a dose dependent decrease in the production of leukotrienes which was statistically significant at 200 mg%. At 1000 mg%, ethanol caused a 20-50% decrease in leukotrienes and a 21% decrease in the amount of prostaglandin D2 (PGD2) formed in these cells. Secondly, when cells were incubated with exogenous arachidonic acid (14 micrograms/ml), large increases in both PGD2 and leukotrienes occurred. Under these conditions, ethanol caused a further increase in the amount of leukotrienes and a small increase in the amount of PGD2 formed. This stimulatory effect was specific for ethanol since neither t-butanol nor n-butanol caused the enhanced production of leukotrienes with exogenous arachidonic acid. Thus, these experiments suggest that ethanol affects metabolism of arachidonic acid at reasonably low doses (200-400 mg%) of ethanol in a manner dependent on the free arachidonic acid in the tissue. Also, in vitro experiments in which ethanol is used as a solvent for arachidonic acid could be greatly affected by high levels of ethanol (500-1000 mg%) which are frequently utilized.

1-Butanol↗
PubMed2006

The alcohol-preferring AA and alcohol-avoiding ANA rats: neurobiology of the regulation of alcohol drinking.

The AA (alko, alcohol) and ANA (alko, non-alcohol) rat lines were among the earliest rodent lines produced by bidirectional selection for ethanol preference. The purpose of this review is to highlight the strategies for understanding the neurobiological factors underlying differential alcohol-drinking behavior in these lines. Most early work evaluated functioning of the major neurotransmitter systems implicated in drug reward in the lines. No consistent line differences were found in the dopaminergic system either under baseline conditions or after ethanol challenges. However, increased opioidergic tone in the ventral striatum and a deficiency in endocannabinoid signaling in the prefrontal cortex of AA rats may comprise mechanisms leading to increased ethanol consumption. Because complex behaviors, such as ethanol drinking, are not likely to be controlled by single factors, system-oriented molecular-profiling strategies have been used recently. Microarray based expression analysis of AA and ANA brains and novel data-mining strategies provide a system biological view that allows us to formulate a hypothesis on the mechanism underlying selection for ethanol preference. Two main factors appear active in the selection: a recruitment of signal transduction networks, including mitogen-activated protein kinases and calcium pathways and involving transcription factors such as Creb, Myc and Max, to mediate ethanol reinforcement and plasticity. The second factor acts on the mitochondrion and most likely provides metabolic flexibility for alternative substrate utilization in the presence of low amounts of ethanol.

Alcohol Drinking↗
PubMed2003

[Toxicity of the ethanol extract of Magonia pubescens on larvae Aedes aegypti].

Crude ethanol extracted from Magonia pubescens trunks was utilized to prove its effect in the intestinal tract of Aedes aegypti 3rd instar larvae. Exposure times (2, 4, 6, 8, 10, 12, and 13 hours) were tested to verify when the morphological alterations begin to occur, in the intestinal tract of the larvae. The toxic effect of the extract was mainly in the midgut, beginning at the anterior midgut and followed through to posterior midgut. The main alterations observed were partial or total cell destruction, high citoplasmatic vacuolization, increase of subperitrophic space, cell hypertrophy and the epithelium did not maintain its monolayer appearance. The alterations began after four hours of exposure to M. pubescens extract. Observations of tissue sections from larvae treated for different lengths of time revealed a wide variation in the degree of damage between exposure periods and midgut larvae region. The present study provides evidence regarding the mode of action of the M. pubescens extract and suggests its potential utilization as a larvicide to control Aedes aegypti mosquito.

Aedes↗
PubMed2000

Factors affecting the biodegradation of cyanide by Burkholderia cepacia strain C-3.

The utilization of cyanide as a nitrogen source for growth in a liquid minimal medium by Burkholderia cepacia strain C-3 isolated from soil was demonstrated. The effects of various parameters on the biodegradation of cyanide by the strain were investigated. Growth at the expense of cyanide occurred over a pH range from 8 to 10, and was optimal at pH 10. Growth and cyanide biodegradation were optimal at 30 degrees C. The presence of Cu2+ or Fe2+ in the liquid medium at a concentration of 1 mM inhibited both the growth of the bacterium and its cyanide degradation. The effects of Ni2+ and Co2+, and to a lesser extent Mn2+ and Mo2+, on cyanide degradation rate were concentration dependent. Addition of the cyanide-containing waste contaminants ethanol and methanol reduced the cyanide utilization of the isolate, but phenol was completely inhibitory. Microbial depletion of cyanide occurred even in the presence of other organic and inorganic nitrogen sources. Fructose, glucose, and mannose were the preferred carbon sources for cyanide biodegradation. The highest rate of cyanide degradation by B. cepacia C-3, 1.85 mg CN h(-1), was observed with 0.25% (w/v) fructose; the degradation rate decreased progressively thereafter with increasing fructose concentration.

Journal Article↗
PubMedDate not supplied

[Glucose consumption by the rat brain in ethanol intoxication and the abstinence syndrome].

The effect of acute and chronic alcohol intoxication and of the syndrome of ethanol withdrawal on the consumption of glucose by the brain of rats was studied by means of intravascular ethanol infusion. Infusion of ethanol into the internal carotid artery had no effect on glucose consumption by the brain, while its infusion into the femoral vein reduced consumption twofold. The effect was completely removed by the inhibitor of alcohol dehydrogenase pyrazole. Chronic intoxication also caused a twofold decrease of glucose utilization by the brain of rats. Infusion of ethanol into the internal carotid artery of rats who were in a state of alcoholic intoxication led to increase of glucose consumption by the brain to the control level. Infusion of ethanol into the femoral vein in this case had no effect on glucose consumption by the rat brain. Utilization of glucose by the brain diminished to an equal degree in rats suffering from the syndrome of ethanol withdrawal and in animals who were in a state of alcoholic intoxication. Infusion of ethanol, both intraarterial and intravenous, had no effect on glucose consumption by the brain. Activation and inhibition of the function of external respiration were encountered in equal concentrations of ethanol in blood flowing from the brain, whatever the method of its infusion.

Alcoholic Intoxication↗
PubMed1999

Ethanol synthesis by genetic engineering in cyanobacteria.

Cyanobacteria are autotrophic prokaryotes which carry out oxygenic photosynthesis and accumulate glycogen as the major form of stored carbon. In this research, we introduced new genes into a cyanobacterium in order to create a novel pathway for fixed carbon utilization which results in the synthesis of ethanol. The coding sequences of pyruvate decarboxylase (pdc) and alcohol dehydrogenase II (adh) from the bacterium Zymomonas mobilis were cloned into the shuttle vector pCB4 and then used to transform the cyanobacterium Synechococcus sp. strain PCC 7942. Under control of the promoter from the rbcLS operon encoding the cyanobacterial ribulose-1, 5-bisphosphate carboxylase/oxygenase, the pdc and adh genes were expressed at high levels, as demonstrated by Western blotting and enzyme activity analyses. The transformed cyanobacterium synthesized ethanol, which diffused from the cells into the culture medium. As cyanobacteria have simple growth requirements and use light, CO2, and inorganic elements efficiently, production of ethanol by cyanobacteria is a potential system for bioconversion of solar energy and CO2 into a valuable resource.

Journal Article↗
PubMed1996

Metabolic mapping of the effects of chronic voluntary ethanol consumption in rats.

The 2-[14C]deoxyglucose method was used to examine the effects of chronic, voluntary ethanol consumption on rates of local cerebral glucose utilization (LCGU). LCGU was measured in male Long-Evans rats immediately following the completion of a 60-min schedule-induced polydipsia drinking session. Three groups of animals were examined: animals with a history of ethanol consumption that received ethanol on the test day (ethanol-ethanol), animals with a similar ethanol history that were presented with water on the test day (ethanol-water), and a control group that received water throughout the experiment (water-water). Ethanol consumption on the test day resulted in a highly discrete pattern of metabolic changes, with significant decreases in glucose utilization in the hippocampal complex, habenula, anterior ventral thalamus, and mammillary bodies, whereas increases were observed in the nucleus accumbens and locus coeruleus. Rates of LCGU in the ethanol-water group were increased throughout all regions of the central nervous system examined, indicating that the long-term consumption of moderate ethanol doses that do not produce physical dependence can cause significant changes in functional brain activity.

Alcohol Drinking↗
PubMedDate not supplied

Alcohol-nutrition interaction (1983).

Alcohol remains a prevailing cause of malnutrition resulting in a variety of deficiency states secondary to decreased intake of nutrients. In addition to various well-described primary malnutrition syndromes, secondary malnutrition may result from the interference of ethanol with nutrient digestion, absorption or utilization. Some of the latter alcohol-nutrient interactions have been recently defined and their pathogenesis is discussed here. Included are interactions with thiamin, folic acid, vitamin A and disorders secondary to amino acid imbalances. The rationale for various forms of therapy is reviewed, including the pitfalls of excess nutrient administration (particularly as it pertains to pyridoxine, vitamin A and amino acids). Desirability of recognizing early precirrhotic stages of alcoholic fibrosis is emphasized in order to start therapy prior to the medical and/or social disintegration of the alcoholic.

Alcoholism↗
PubMed1983

Ethanol-induced male reproductive tract pathology as a function of ethanol dose and duration of exposure.

Chronic ethanol ingestion by the male results in several manifestations of reproductive dysfunction. However, little is known concerning 1) the level of ethanol or duration of ethanol exposure necessary to produce pathological changes in the male reproductive tract; or 2) mechanism(s) by which ethanol exerts its toxic effects. The purpose of the present study was to develop an animal model which describes ethanol-induced male reproductive failure and to define conditions of ethanol exposure which alter male reproductive tract function. Proven breeder male C57Bl/6J mice were maintained on either a 5% (v/v) ethanol liquid diet for periods of 5, 10 or 20 weeks or a 6% (v/v) ethanol diet for 5 weeks. Daily blood ethanol levels peaked at 125 to 200 and 268 mg/100 ml in animals consuming the 5 and 6% ethanol diets, respectively. Plasma testosterone levels were depressed throughout all treatments, an effect which appeared reversible. Plasma testosterone was poorly correlated (r = 0.28) with blood ethanol levels during treatment. After treatment, reproductive tract pathology included reduced testicular and accessory sex organ weights, decreased spermatogenesis, increased desquamation of immature germ cells into the seminiferous tubule lumina and increased frequency of inactive seminiferous tubules. Whereas minimal changes were noted after 5 weeks treatment with the 5% ethanol diet, deficiencies became increasingly evident after either longer exposure periods or treatment with 6% ethanol for the same period. None of the treatment regimens resulted in altered plasma levels of hepatic enzyme markers or in altered histological appearance. The data indicate that male reproductive function is sensitive to both ethanol dose and duration of exposure. The animal model described in the present study should have utility in the evaluation of putative mechanisms of ethanol-induced male reproductive dysfunction.

Alcoholism↗
PubMed1983

Preparation and stabilization of aqueous/ethanolic superoxide solutions.

Methods for the generation and stabilization of alkaline aqueous and/or ethanolic superoxide solutions have been developed utilizing either high energy ionizing radiation or vacuum-uv photolysis. These solutions can be used for the study of the reactivity of superoxide radical (O-2) and its conjugate acid the perhydroxyl radical (HO2) with other compounds over the entire pH range. Frozen, they can be kept at -196 degrees C for prolonged time periods with little loss in superoxide radical. As these solutions can be rendered free of molecular oxygen, studies of the reactivity of HO2/O-2 with oxygen-sensitive compounds under virtually anaerobic conditions are possible.

Chemical Phenomena↗