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Isotonic but not hypertonic ethanol stimulates LHRH secretion from perifused rat median eminence.

We utilized luteinizing hormone-releasing hormone (LHRH) secretion by perifused minced rat hypothalamic median eminence (ME) tissue to evaluate whether isotonic ethanol would stimulate neurosecretion as it does secretion from pituitary cells. Isotonic ethanol induced a dose-dependent burst of LHRH secretion which was maximal at 2-3 min and returned to near baseline by 10 min. In Ca(2+)-depleted media (< 2 microM Ca2+), stimulation of secretion by isotonic ethanol was enhanced, but secretion induced by depolarizing 30 mM K+ was abolished. Hypertonic ethanol was ineffective in stimulating LHRH secretion in either normal or Ca(2+)-free media. The secretory response of hypothalamic LHRH-secreting cells to ethanol and its negative modulation by medium Ca2+ is thus identical to that of normal anterior pituitary cells and presumably is caused by cell swelling resulting from influx of permeant ethanol molecules across the plasmalemma.

Animals↗

Origin of hepatic triglyceride fatty acids: quantitative estimation of the relative contributions of linoleic acid by diet and adipose tissue in normal and ethanol-fed rats.

The present study demonstrates that the rat liver obtains most of its triglyceride fatty acids from dietary sources. The dietary and adipose tissue contributions of linoleic acid for hepatic triglyceride esterification were shown to be 50.42 and 13.85 micro moles, respectively, during a 4-day period. When ethanol provided 40% of the caloric intake, fatty liver developed and hepatic triglyceride content increased threefold. Under these conditions, the dietary and adipose tissue contributions of linoleic acid were estimated at 192.85 and 10.73 micro moles, respectively. This increase in dietary fatty acid utilization was sufficient to account for the entire increase in esterified hepatic linoleic acid. Any explanation of these observations must include the high dietary fatty acid utilization in both control and ethanol-treated animals. One possibility is that most dietary lipids first enter a rapidly turning over pool in adipose tissue from which most hepatic triglyceride fatty acids are derived. Another is that dietary fatty acids, incorporated into chylomicrons, are stored separately and used preferentially by the liver as compared with lipids derived from adipose tissue and bound to albumin. The pros and cons of these possibilities are discussed.

Adipose Tissue↗

Gas-liquid chromatographic analysis of ethanol and acetaldehyde in blood with minimal artifactual acetaldehyde formation.

A gas-liquid chromatographic procedure utilizing headspace gas analysis is described for the determination of ethanol and its metabolite, acetaldehyde, in a 100-microliters sample of blood from the rat, guinea pig, sheep, or human. Artifactual formation of ethanol-derived acetaldehyde is minimized during sample preparation by using a chemical solution containing perchloric acid and sodium azide in saline, and thiourea. Aqueous standards of ethanol and acetaldehyde are used to calibrate the procedure, and 1-propanol is used as the internal standard of the method. The recovery of ethanol and acetaldehyde from spiked blood samples is quantitative and reproducible, with a within-day coefficient of variation of less than 7% for ethanol and less than 9% for acetaldehyde. The lower limit of quantitative sensitivity is 0.006 mg/ml ethanol and 0.10 microgram/ml acetaldehyde. The instrumental analysis time is less than 3 min, which enables high sample throughput.

Acetaldehyde↗

Chronic ethanol administration alters hepatic rates of glycerol phosphorylation and glycerol 3-phosphate oxidation: a dynamic in vivo 31P magnetic resonance spectroscopy study.

We used dynamic in vivo 31P magnetic resonance spectroscopy to noninvasively study the metabolism of glycerol by the liver in living rats, as a means of detecting subtle metabolic changes induced by chronic ethanol consumption. Rats subjected to chronic ethanol consumption and their pair-fed controls were given a metabolic load of glycerol (0.75 or 1.3 mL glycerol x kg body mass(-1), i.p. or i.v) under normoxic or hyperoxic (98% O2) conditions. Changes in the level of glycerol 3-phosphate were followed in situ by monitoring the hepatic 31P phosphomonoester resonance every 7 or 13 min for up to 330 min. When challenged with a large dose of glycerol, chronic ethanol-treated rats exhibited less accumulation of glycerol 3-phosphate than controls, independent of the route of administration of the glycerol or whether the two groups were fasted or fed. For example, 1.3 mL glycerol x kg(-1) i.v. under normoxic conditions resulted in a two-fold increase in phosphomonoester in ethanol-treated rats compared with a five-fold increase in controls. The ethanol-treated rats also showed a slower rate of phosphorylation of glycerol and slower oxidation of glycerol 3-phosphate than controls, indicating decreased activities of the glycerol kinase and glycerol 3-phosphate dehydrogenase steps, and hence slower glycerol utilization. The rate of glycerol utilization was dose and oxygen concentration dependent. Kinetic analysis indicated that the chronic ethanol-induced decrease in the glycerol 3-phosphate dehydrogenase reaction was due to a decreased rate of NADH reoxidation in the liver, likely owing to a decrease in oxygen supply or utilization in the ethanol-treated rats. These observations support the hypothesis of pre-existing hypoxia in rat liver after chronic ethanol administration. This study demonstrates the utility of dynamic in vivo 31P magnetic resonance spectroscopy in following the metabolism of a glycerol load as a sensitive, nonperturbing, and potentially clinically applicable test of liver function.

Adenosine Triphosphate↗

Effects of heat and alcohol treatments of soybean meal on nitrogen utilization by sheep.

Soybean meal (SBM) was treated with aqueous solutions of ethanol or propanol at room temperature or at 80 C to study treatment effects on SBM-N solubility and utilization by sheep. Soybean meal was soaked in an excess of 70% (v/v) ethanol at 80 C (ET-80), 70% ethanol at 23 C (ET-23) or 70% propanol at 80 C (PR-80). Nontreated SBM and nontreated SBM heated at 80 C without alcohol treatment (NT-80) served as controls. Nitrogen solubility in McDougall's buffer was lowest (P less than .05) for PR-80 and ET-80 (2.2 and 4.7% of total N, respectively), intermediate (P less than .05) for ET-23 (9.0%), greater (P less than .05) for nontreated SBM (36.2%) and highest for NT-80 (40.2%). In an situ study using three ruminally cannulated cows and two bags per treatment per animal per removal time, more (P less than .05) N remained in in situ bags after 3, 6, 9 and 12 h incubation for ET-23, ET-80 and PR-80 than for nontreated SBM and NT-80. A lamb metabolism trial, using 15 lambs in each of two periods, compared nontreated SBM, ET-23, ET-80, PR-80 and urea as N supplements. Nitrogen retention was higher (P less than .02) for lambs fed SBM treatments compared with urea. When the same N supplements were fed to wethers in a 5 X 5 Latin square experiment and duodenal N flow was measured, non-ammonia non-bacterial N flow was higher (P less than .07) for wethers fed SBM treatments than for wethers fed urea.(ABSTRACT TRUNCATED AT 250 WORDS)

1-Propanol↗

Central nucleus of the amygdala and the effects of alcohol and alcohol-drinking behavior in rodents.

This article will review key literature on the effects of alcohol on the amygdala and the involvement of the amygdala in regulating alcohol drinking in mice and rats. Special emphasis will be placed on the central nucleus of the amygdala (CeA) because this nucleus is a major component of the extended amygdala, which has been implicated in regulating alcohol-drinking behavior. Immunocytochemical and in situ hybridization studies indicate that acute high-dose ethanol administration increases c-fos expression in GABAergic neurons within the CeA of the rat, suggesting activation of these neurons by ethanol. A similar high-dose (4 g/kg ethanol) effect on c-fos expression in the CeA of C57 mice was also observed, whereas the DBA mice showed increased c-fos expression in the CeA in the dose range of 1.25-4.0 g/kg. Studies with DBA x C57 F2 intercross mice suggest that there may be a relationship between the neuronal activating effects of ethanol in the CeA and the locomotor stimulating effects of ethanol. Studies with rats examining the effects of acute ethanol or chronic alcohol drinking on local cerebral glucose utilization (LCGU) rates (as a measure of synaptic activity) indicated that (a) acute ethanol (0.25-2.0 g/kg) had little effect on LCGU rates in the CeA; (b) basal LCGU rates were reduced in the CeA as a result of chronic alcohol drinking; and (c) oral self-administration of ethanol increased LCGU values within the CeA. Microdialysis studies demonstrated that acute ethanol (2 g/kg) injection increased dopamine (DA) and serotonin (5-HT) release in the CeA. Microinjection studies indicate that GABA(A) receptors within the CeA are involved in oral ethanol self-administration. Overall, the findings from the various studies support a role for the CeA in mediating the stimulating actions of alcohol in mice and regulating alcohol-drinking behavior in mice and rats.

Alcohol Drinking↗

Acute effects of ethanol on the perfused rat liver. Studies on lipid and carbohydrate metabolism, substrate cycling and perfusate amino acids.

1. Livers from fed rats were perfused in situ with whole rat blood containing glucose labelled uniformly with (14)C and specifically with (3)H at positions 2, 3 or 6. 2. When ethanol was infused at a concentration of 24mumol/ml of blood the rate of utilization was 2.8mumol/min per g of liver. 3. Ethanol infusion raised perfusate glucose concentrations and caused a 2.5-fold increase in hepatic glucose output. 4. Final blood lactate concentrations were decreased in ethanol-infused livers, but the mean uptake of lactate from erythrocyte glycolysis was unaffected. 5. Production of ketone bodies (3-hydroxybutyrate+3-oxobutyrate) and the ratio [3-hydroxybutyrate]/[3-oxobutyrate] were raised by ethanol. 6. Formation of (3)H(2)O from specifically (3)H-labelled glucoses increased in the order [6-(3)H]<[3-(3)H]<[2-(3)H]. Production of (3)H(2)O from [2-(3)H]glucose was significantly greater than that from [3-(3)H]glucose in both control and ethanol-infused livers. Ethanol significantly decreased (3)H(2)O formation from all [(3)H]glucoses. 7. Liver glycogen content was unaffected by ethanol infusion. 8. Production of very-low-density lipoprotein triacylglycerols was inhibited by ethanol and there was a small increase in liver triacylglycerols. Very-low-density-lipoprotein secretion was negatively correlated with the ratio [3-hydroxybutyrate]/[3-oxobutyrate]. Perfusate fatty acid concentrations and molar composition were unaffected by perfusion with ethanol. 9. Ethanol decreased the incorporation of [U-(14)C]glucose into fatty acids and cholesterol. 10. The concentration of total plasma amino acids was unchanged by ethanol, but the concentrations of alanine and glycine were decreased and ([glutamate]+[glutamine]) was raised. 11. It is proposed that the observed effects of ethanol on carbohydrate metabolism are due to an increased conversion of lactate into glucose, possibly by inhibition of pyruvate dehydrogenase. The increase in gluconeogenesis is accompanied by diminished substrate cycling at glucose-glucose 6-phosphate and at fructose 6-phosphate-fructose 1,6-bisphosphate.

Amino Acids↗

Effects of the Turkish subspecies of Viscum album on macrophage-derived cytokines.

Viscum album L. herbs are used for the treatment of various diseases. Apart from their immunostimulatory, antitumor, and hypotension inducing activities, anti-inflammatory effects has also been recorded in the literature. Since the immunostimulatory activity of mistletoe is mainly attributed to a stimulation of the mononuclear phagocytic system and to an induction of inflammation by macrophage-derived cytokines, utilization based on anti-inflammatory activity seems in contrast with its use as immunostimulant. The inhibitory effects of 80% ethanol extracts and subfractions obtained by petroleum ether, diethylether, ethylacetate and n-butanol from three Turkish subspecies of V. album L. (ssp. album, ssp. abietis, ssp. austriacum) against interleukins (IL-1alpha and IL-1beta) and tumor necrosis factor-alpha were studied. Ethanolic (80%) extracts of all three subspecies exhibited almost no significant inhibitory activity on the proinflammatory cytokines, at least in the concentrations applied in this study. In contrast, they induce a concentration dependent stimulation of these cytokines. On the other hand, moderate to weak, dose dependent inhibitory effects were observed on fractionation, which may be attributed to the existence of less polar components with anti-inflammatory activity. Based on our results, it was concluded that the utilization of ethanol extracts for anti-inflammatory purposes would not be beneficial probably due to the partial extraction of the low-molecular-weight polypeptides, i.e. viscotoxins, which induce an inflammatory response. Moreover, for anti-inflammatory effects, extracts of Viscum album ssp. austriacum may be preferred, since higher inhibitory ratios were obtained against inflammatory cytokines than with the other two subspecies.

Dose-Response Relationship, Drug↗

Effect of ethanol on nucleolar structure: a cytological indication of change in RNA/protein synthesis.

The effects of chronic ethanol ingestion on an organelle known to be involved in protein synthesis were studied cytologically in nerve cells of the adult hamster. Twenty-six animals were administered standard laboratory chow and either tap water (controls) or a 15% ethanol solution (experimentals) for a period of 7 weeks. Brains were perfusion-fixed, sectioned transversely, and stained with buffered thionin for microscopic analysis. Reported here are changes in an RNA-rich intranucleolar body (INB) seen in facial motor neurons and cerebellar Purkinje cells of the golden hamster. After chronic ethanol ingestion, the size and frequency of the INB increased significantly in both cell populations. Theoretical considerations are discussed concerning the correlation between this apparent storing of nucleolar RNA/RNP and the biochemical evidence of other investigators for ethanol-induced alterations in RNA/protein synthesis and utilization in neurons.

Animals↗

trans-dominant mutations in the GPR1 gene cause high sensitivity to acetic acid and ethanol in the yeast Yarrowia lipolytica.

Acetate non-utilizing strains harbouring trans-dominant mutations in the GPR1 gene (GPR1(d)) of the dimorphic yeast Yarrowia lipolytica have been selected and characterized. These mutants are highly sensitive to low concentrations of acetic acid and ethanol, even in presence of glucose. The toxic effect of acetic acid is pH-dependent and has the strongest effect at low pH. In contrast, the action of ethanol is pH-independent. One GPR1(d) mutant has been detected that was highly sensitive to acetic acid but could still grow on ethanol, which indicates putative differences in the function of the GPR1 gene product in the sensitivity to acetic acid and ethanol. The GPR1(d) mutants exhibit a complex pleiotropic phenotype. The mutations cause changed colony morphology as well as dimorphism of cells, and induce early cell death during growth on glucose, even without the presence of dicarbon compounds. Composition of intracellular membranes, as well as morphology of vacuole and mitochondria, were strongly changed. Back-crosses with wild-type strains and analysis of recombinant strains have shown that the expression of the pleiotropic phenotype depends on the site of mutation in the GPR1 gene, as well as on the genetic background of the strain harbouring the responsive mutation. Our data suggest that Gpr1p is involved in a general response of cells to the toxic action of dicarbon compounds like acetic acid and ethanol.

Acetic Acid↗

Kinetic analysis of ethanol production from glucose fermentation by yeast cells immobilized onto ceramic supports.

Porous ceramic supports have been developed and utilized for the immobilization of yeast cells to produce ethanol by the fermentation of glucose. The relationship between the porous structure of the ceramic support and the quantity of yeast cells immobilized and the production of ethanol by the fermentation have been investigated. A comparison of the properties of the ceramic supports with those of a calcium alginate gel indicated that the ceramics are the better of the two types of material and have potential for industrial application.

Absorption↗

Adaptations in adenosine signaling in drug dependence: therapeutic implications.

Adenosine is an important endogenous purine neuromodulator in the central nervous system that modulates many important cellular processes in neurons. The physiological effects of adenosine are transduced through four pharmacologically classified receptor types i.e., A1, A2A, A2B and A3. All adenosine receptors are G-protein coupled receptors (GPCR) of the type 1 variety. Adaptations in adenosine signaling have been implicated in a wide range of pathophysiological processes, such as epilepsies, sleep disorders, pain, and drug addictions. Knowledge relating to the etiology of addictive processes is far from complete, and as a result the therapeutic options to deal with drug dependence issues are limited. Drugs of abuse mediate their effects through many distinct cellular effectors, such as neurotransmitter transporters, ion channels, and receptor proteins. However, a unifying feature of the major drugs of abuse-i.e., opiates, cocaine, and alcohol-is that they all directly or indirectly modulate adenosine signaling in neurons. Agents targeting adenosine receptors may therefore offer novel avenues for the development of therapies to manage or treat addictions. A consistent cellular adaptation to long-term drug use is the up- or down-regulation of signaling pathways driven by adenylyl cyclase/cyclic AMP (cAMP) in several brain regions linked to addiction. Withdrawal from mu-opioids or cocaine following their chronic administration leads to an upregulation of adenylyl cyclase-mediated signaling, resulting in high levels of cAMP. Cyclic AMP produced in this way acts as a substrate for the endogenous production of adenosine. Increased levels of endogenous adenosine interact with presynaptic A1 receptors to inhibit the excessive neuronal excitation often seen during morphine/cocaine withdrawal. These pre-clinical findings fit well with other data indicating that drugs which boost endogenous adenosine levels or directly interact with inhibitory A1 receptors can alleviate many of the negative consequences of opioid/cocaine withdrawal. Ethanol interacts directly with the adenosine system by blocking nucleoside transporters in the cell membrane. The effect of this inhibition is an increase in extracellular adenosine levels and adenosine receptor activation. Depending on the time course of ethanol exposure and the receptor population present, cAMP levels are either reduced or increased. Chronic ethanol treatment tends to reduce cAMP levels as a consequence of the desensitization of stimulatory GPCRs (such as A2-type receptors) seen following prolonged receptor activation. Unlike opiates and cocaine, adenosine receptor activation worsens the behavioral effects of drug ingestion, and evidence indicates that agents that negatively modulate adenosine receptor function have some utility in attenuating the effects of ethanol use. Taken together, these data suggest that pharmacological manipulation of adenosine signaling represents a potentially useful means of managing drug dependence.

Adenosine↗

I. Functional consequences of intragastrically administered ethanol in rats as measured by the 2-[14C]deoxyglucose method.

The 2-[14C] deoxyglucose (2DG) method was used to examine the effects of the acute, intragastric (IG) administration of ethanol on rates of local cerebral glucose utilization (LCGU) in male Sprague-Dawley rats. The 2DG technique was initiated 3 min after the administration of 0.25, 1.0, or 2.0 g/kg ethanol. Each dose of ethanol examined produced a discrete pattern of change in functional activity. The administration of 0.25 g/kg ethanol increased LCGU in structures of the mesocorticolimbic and nigrostriatal circuits. The administration of 1.0 and 2.0 g/kg ethanol, however, produced a more limited pattern of changes in LCGU, with increases observed primarily in the mesocorticolimbic circuit. The increases in functional activity observed after the administration of the 0.25 g/kg ethanol dose were more widespread, and generally of larger magnitude, as compared to the 1.0 and 2.0 g/kg ethanol doses. These data indicate that structures of the mesocorticolimbic system are particularly sensitive to the effects of both low and high doses of ethanol when the effects are measured on the ascending limb of the blood ethanol curve.

Administration, Oral↗

An unusual oxygen-sensitive, iron- and zinc-containing alcohol dehydrogenase from the hyperthermophilic archaeon Pyrococcus furiosus.

Pyrococcus furiosus is a hyperthermophilic archaeon that grows optimally at 100 degreesC by the fermentation of peptides and carbohydrates to produce acetate, CO2, and H2, together with minor amounts of ethanol. The organism also generates H2S in the presence of elemental sulfur (S0). Cell extracts contained NADP-dependent alcohol dehydrogenase activity (0.2 to 0.5 U/mg) with ethanol as the substrate, the specific activity of which was comparable in cells grown with and without S0. The enzyme was purified by multistep column chromatography. It has a subunit molecular weight of 48,000 +/- 1,000, appears to be a homohexamer, and contains iron ( approximately 1.0 g-atom/subunit) and zinc ( approximately 1.0 g-atom/subunit) as determined by chemical analysis and plasma emission spectroscopy. Neither other metals nor acid-labile sulfur was detected. Analysis using electron paramagnetic resonance spectroscopy indicated that the iron was present as low-spin Fe(II). The enzyme is oxygen sensitive and has a half-life in air of about 1 h at 23 degreesC. It is stable under anaerobic conditions even at high temperature, with half-lives at 85 and 95 degreesC of 160 and 7 h, respectively. The optimum pH for ethanol oxidation was between 9. 4 and 10.2 (at 80 degreesC), and the apparent Kms (at 80 degreesC) for ethanol, acetaldehyde, NADP, and NAD were 29.4, 0.17, 0.071, and 20 mM, respectively. P. furiosus alcohol dehydrogenase utilizes a range of alcohols and aldehydes, including ethanol, 2-phenylethanol, tryptophol, 1,3-propanediol, acetaldehyde, phenylacetaldehyde, and methyl glyoxal. Kinetic analyses indicated a marked preference for catalyzing aldehyde reduction with NADPH as the electron donor. Accordingly, the proposed physiological role of this unusual alcohol dehydrogenase is in the production of alcohols. This reaction simultaneously disposes of excess reducing equivalents and removes toxic aldehydes, both of which are products of fermentation.

Alcohol Dehydrogenase↗

Glycyl-glutamine in nucleus accumbens reduces ethanol intake in alcohol preferring (P) rats.

Opioid peptides and glycyl-glutamine (Gly-Gln) have been implicated in the control of ethanol consumption. A recognized beta-endorphin cleavage product, Gly-Gln, inhibits voluntary alcohol consumption when microinjected into the nucleus accumbens (AcbSh) of P rats. To evaluate the site-specific efficacy of Gly-Gln on ethanol consumption following AcbSh application, ethanol preferring (P) rats were allowed to establish individual baseline ethanol/water consumption utilizing a voluntary self-administration paradigm. Subsequent to baseline ethanol consumption being established, bilateral guide cannulae were stereotaxically implanted +1 mm dorsal to the AcbSh for subsequent Gly-Gln (100 nmol/microl) or saline vehicle (1 microl) injections. Alcohol intake, body weight, and water intake were measured at 24 h post-injection intervals. Unilateral Gly-Gln injections reduced ethanol consumption 35.6% (P < 0.05) from pre-established baseline consumption (6.24 +/- 0.64 g/kg to 4.06 +/- 0.28 g/kg). Bilateral Gly-Gln injections further reduced consumption to 51.9% (6.4 +/- 1.0 g/kg to 3.08 +/- 0.65 g/kg at 24 h (P < 0.01) below established baseline values within 24 h without significant changes in body weight or water consumption. Also, the amino acid constituents of the dipeptide had no influence on ethanol consumption behavior; however, Gly-Gln efficacy was shown to be comparable to central beta-endorphin-(1-27) or intraperitoneal (i.p.) naltrexone-induced suppression of ethanol intake. These data indicate that the AcbSh exhibits a site-specific sensitivity to the suppressive actions of Gly-Gln or beta-endorphin-(1-27) injections that modulate voluntary ethanol consumption in P rats. These findings support the broader concept that select forebrain opioid-responsive neural sites may influence the development or expression of alcohol abuse syndromes in animal models or humans.

Alcohol-Induced Disorders, Nervous System↗

Evaluation of ethanol-fixed, paraffin-embedded tissues for proteomic applications.

We previously reported that ethanol fixation and paraffin embedding of tissues produce excellent histomorphology and good preservation of macromolecules. Here, we present a detailed evaluation of ethanol-fixed tissues for proteomic initiatives. When proteins were extracted from ethanol-fixed, paraffin-embedded prostate tissue, resolved by two-dimensional gel electrophoresis (2-DE), and stained by standard methods, several hundred protein molecules could be detected and successfully analyzed by mass spectrometry. Protein profiles obtained from ethanol-fixed tissues were highly similar to those observed from frozen tissues, in contrast to the poor protein recovery from formalin-fixed material. The protein content of specific cells that were microdissected from ethanol-fixed tissue sections using laser capture microdissection could also be successfully analyzed by 2-DE. We observed that eosin staining of tissue sections had a detrimental effect on protein separation, whereas hematoxylin staining had minimal consequence. In order to illustrate the applicability of ethanol-fixed tissues for proteomic discovery studies, we compared the protein profiles of patient-matched, normal prostatic epithelial cells and invasive adenocarcinoma cells obtained from ethanol-fixed, paraffin-embedded tissues. A number of differentially expressed proteins was discovered and identified by mass spectrometry. Immunohistochemical analyses performed on ethanol-fixed tissue sections were in agreement with the proteomic discovery findings. In light of these results, we conclude that ethanol-fixed tissues can be successfully utilized for proteomic analyses.

Electrophoresis, Gel, Two-Dimensional↗

Construction of a xylan-fermenting yeast strain through codisplay of xylanolytic enzymes on the surface of xylose-utilizing Saccharomyces cerevisiae cells.

Hemicellulose is one of the major forms of biomass in lignocellulose, and its essential component is xylan. We used a cell surface engineering system based on alpha-agglutinin to construct a Saccharomyces cerevisiae yeast strain codisplaying two types of xylan-degrading enzymes, namely, xylanase II (XYNII) from Trichoderma reesei QM9414 and beta-xylosidase (XylA) from Aspergillus oryzae NiaD300, on the cell surface. In a high-performance liquid chromatography analysis, xylose was detected as the main product of the yeast strain codisplaying XYNII and XylA, while xylobiose and xylotriose were detected as the main products of a yeast strain displaying XYNII on the cell surface. These results indicate that xylan is sequentially hydrolyzed to xylose by the codisplayed XYNII and XylA. In a further step toward achieving the simultaneous saccharification and fermentation of xylan, a xylan-utilizing S. cerevisiae strain was constructed by codisplaying XYNII and XylA and introducing genes for xylose utilization, namely, those encoding xylose reductase and xylitol dehydrogenase from Pichia stipitis and xylulokinase from S. cerevisiae. After 62 h of fermentation, 7.1 g of ethanol per liter was directly produced from birchwood xylan, and the yield in terms of grams of ethanol per gram of carbohydrate consumed was 0.30 g/g. These results demonstrate that the direct conversion of xylan to ethanol is accomplished by the xylan-utilizing S. cerevisiae strain.

Base Sequence↗

Larval responses to environmental ethanol in Drosophila melanogaster: variation within and among populations.

There is a latitudinal cline in attraction to ethanol of newly hatched D. melanogaster larvae; attraction decreases as one moves from the temperature regions to the Australian tropics. Tropical populations manifest high levels of heterogeneity compared with temperate regions, making tropical populations less dependent on ethanol. Since ethanol is a resource for D. melanogaster, an approach to the genetics of resource utilization in natural populations via behavior genetics of larval ethanol responses is feasible, using isofemale strains as experimental material.

Analysis of Variance↗