Ethanol Supplementation Research Articles

Effect of tween 40 and ethanol on the secretion, structure and antioxidant activities of exopolysaccharides from Inonotus rickii.

In this study, the effects of Tween 40 and ethanol supplementation on the secretion, structure and antioxidant activities of exopolysaccharide (EPS) from Inonotus rickii were investigated. It was observed that Tween 40 and ethanol displayed a stimulatory effect on EPS secretion. The EPSs obtained by the addition of Tween 40 (EPS-T), ethanol (EPS-E) and control (EPS-C) were purified by Sepharose CL-6B gel chromatography and molecular weights of EPS-T, EPS-E and EPS-C were estimated to be 22.1, 30.0, and 40.5 kDa, respectively. Monosaccharide composition analysis indicated that EPS-T, EPS-E and EPS-C were mainly composed of mannose and glucose. Furthermore, EPS-E exhibited better OH• and DPPH scavenging activities than those of EPS-C and EPS-T, which might be associated with its molecular characterization.

Single and simultaneous effects of acrylamide and ethanol on bone microstructure of mice after one remodeling cycle

BackgroundThis study aimed to examine femoral bone microstructure of mice after single and simultaneous administration to acrylamide and ethanol since both substances are often consumed separately and/or together by humans. Interactive effects of these toxins were analysed after one remodeling cycle.MethodsTwenty clinically healthy adult mice were randomly divided into four groups following 2 weeks administration of toxins: A group - mice were fed with acrylamide (40 mg/kg bw); E group - mice were ethanol-fed (15% ethanol); AE group - mice were simultaneously fed with both toxins, and a C group – control (without acrylamide and/or ethanol supplementation). Generally, 2D and 3D imaging methods were used to determine cortical and trabecular bone tissues microstructure. Biochemical analyses of plasma parameters were also realized using commercially available ELISA tests and spectrophotometrically.ResultsSingle and simultaneous exposure to acrylamide and ethanol affected only cortical bone microstructure. No significant changes in trabecular bone morphometry were detected among all groups. In mice from the A group, increased endocortical remodeling associated with a higher level of serum calcium and vasoconstriction of primary osteon’s vascular canals (POVC) were identified. On the contrary, increased cortical porosity consistent with a decreased relative bone volume, bone mineral density (BMD) and lower levels of alkaline phosphatase (ALP), glutathione (GSH), calcium in plasma and also with vasodilation of POVC were observed in the E group. In the AE group, the highest density of secondary osteons associated with a lower BMD and decreased levels of ALP, GSH were documented. The parameters of POVC and Haversian canals approximated to the C group. In addition, single and simultaneous exposure to both toxins caused liver disease consistent with a higher values of alanine aminotransferase (ALT), aspartate aminotransferase (AST) in plasma of all experimental groups.ConclusionsSingle administration to acrylamide and ethanol had negative effects on cortical bone structure of mice after one remodeling cycle. However, we identified possible antagonistic impact of these toxins on the structure of the cortical bone.

Effect of ethanol supplementation on the transcriptional landscape of bionanocellulose producer Komagataeibacter xylinus E25

Ethanol exerts a strong positive effect on the cellulose yields from the widely exploited microbial producers of the Komagataeibacter genus. Ethanol is postulated to provide an alternative energy source, enabling effective use of glucose for cellulose biosynthesis rather than for energy acquisition. In this paper, we investigate the effect of ethanol supplementation on the global gene expression profile of Komagataeibacter xylinus E25 using RNA sequencing technology (RNA-seq). We demonstrate that when ethanol is present in the culture medium, glucose metabolism is directed towards cellulose production due to the induction of genes related to UDP-glucose formation and the repression of genes involved in glycolysis and acetan biosynthesis. Transcriptional changes in the pathways of cellulose biosynthesis and c-di-GMP metabolism are also described. The transcript level profiles suggest that Schramm-Hestrin medium supplemented with ethanol promotes bacterial growth by inducing protein biosynthesis and iron uptake. We observed downregulation of genes encoding transposases of the IS110 family which may provide one line of evidence explaining the positive effect of ethanol supplementation on the genotypic stability of K. xylinus E25. The results of this study increase knowledge and understanding of the regulatory effects imposed by ethanol on cellulose biosynthesis, providing new opportunities for directed strain improvement, scaled-up bionanocellulose production, and wider industrial exploitation of the Komagataeibacter species as bacterial cellulose producers.

Low temperature nutrient removal from municipal wastewater by purple phototrophic bacteria (PPB)

This study evaluated the performance of purple phototrophic bacteria (PPB) at temperatures of 22 °C and 11 °C in a membrane photobioreactor. PPB performance and Monod parameters were evaluated at 11 °C and were compared to PPB performance at 22 °C. At 22 °C, the optimum HRT with respect to meeting target limits of TCOD < 50 mg/L, TN < 10 mg/L, and TP < 1 mg/L was 9 h with ethanol supplementation at 300 mgCODL−1 noting that ethanol was only needed to meet the TN limit. However, at 11 °C, the photobioreactor achieved effluent TCOD, TN, and TP concentrations of 70 mg/L, 10 mg/L, and 1.2 mg/L respectively at 9 h HRT and 300 mgCOD/L ethanol addition. Monod kinetic parameters Ks, K, Y, and Kd were determined to be 20 mgCOD/L, 1.8 mgCOD/(mgVSS.d), 0.54 mgVSS/mgCOD, and 0.09 d−1 at 22 °C and 31 mgCOD/L, 1.2 mgCOD/(mgVSS.d), 0.51 mgVSS/mgCOD, and 0.07 d−1 at 11 °C respectively.

&lt;i&gt;In vivo&lt;/i&gt; influence of S-adenosylmethionine supplementation on ethanol teratogenicity in the rat

Studies have shown that maternal alcohol consumption during pregnancy negatively impacts on prenatal development. Ethanol-induced alteration in S-adenosylmethionine (SAMe) metabolism and subsequent depletion of SAMe may be a mechanism involved in fetal ethanol effects. The aim of the present study was to determine the teratogenicity of ethanol and to investigate protective effects of SAMe. Pregnant Sprague-Dawley rats, weighing 220-250 g were used. Pregnant animals were treated from day 6 through 12 of gestation with ethanol alone (6.4% ethanol) or ethanol plus SAMe (200 mg/kg). The groups for the day-20 and day-12 experiments included: Group I (ethanol liquid diet: unrestricted-fed); Group II (isocaloric maltose-dextrin liquid diet: restricted-fed); and Group III (ethanol liquid diet + SAMe: unrestricted-fed). Embryos and fetuses were recovered on gestational day-12 or day-20, respectively, and were quantitatively and qualitatively assessed for developmental anomalies. The data on continuous variables were analyzed using paired T-Test and categorical embryonic development data were processed using Chi-Square Analysis. In the present study, prenatal growth and developmental retardations such as significantly reduced crown-rump length, anomalies of the central nervous system, the heart, limbs and the skeleton were observed in embryos and fetuses of ethanol treated rats. SAMe supplementation of the ethanol treated pregnant rats significantly improved litter weight, from 20.9 + 1.6 g to 23.6 + 0.8 g, and crown-rump length from 25.8 + 0.8 mm to 27.6 + 0.4 mm, and decreased the incidence of resorptions from 1.9 + 0.6 to 0.5 + 0.8 in day-20 fetuses. It also significantly increased embryonic protein content from 173 + 36 μg to 251 + 29 μg and significantly improved the development of brain vesicles and closure of the posterior neuropore in day-12 embryos. The present study clearly showed that ethanol is teratogenic in vivo . SAMe supplementation of ethanol treated pregnant rats partially ameliorated the teratogenic effects of ethanol. This indicated that the observed ethanol related growth and developmental abnormalities may have been, at least partly, due to ethanol-induced SAMe deficiency. Further studies should consider extending the treatment period to the entire gestational period instead of limiting it to only the days covering the period of organogenesis. Keywords: ethanol teratogenicity, Sprague-Dawley rats, S-adenosylmethionine, embryos, fetuses

Neuroprotective effect of Annona glabra extract against ethanol-induced apoptotic neurodegeneration in neonatal rats

The present study aimed to investigate the neuroprotective effect of Annona glabra extract (AGE) against ethanol-induced neurodegeneration in neonatal rats. AGE is known to contain various pharmacological and therapeutic properties. Phytochemical analysis of AGE was performed to understand the presence of vital therapeutic components. Neonatal rats were assigned to the following groups: group I (normal control rats receiving normal saline), group II (control rats receiving ethanol), and group III (treated rats receiving ethanol-AGE). The lipid peroxidation, reduced glutathione (GSH), glutathione peroxidase (Gpx), superoxide dismutase (SOD), catalase, and acetylcholine esterase (AChE) levels were determined. Behavioral parameters, histological features, neuronal cell viability, and apoptosis were also investigated. The presence of flavonoids, terpenoid, glycosides, steroids, saponins, tannins, anthraquinones, and acidic compounds was noted in the AGE. Ethanol supplementation drastically increased the malondialdehyde (MDA) content to 52.17 nmol/g in the control rats (group II). However, the MDA content was reduced to 27.34 nmol/g in ethanol-AGE-treated neonatal rats (group III) compared with control rats. The GSH content was substantially reduced, to 33.68 mg/g, in control rats compared with in normal control rats. However, the GSH content was significantly increased, to 59.32 mg/g, following ethanol-AGE supplementation. Gpx, SOD, catalase, and AChE enzyme activities were increased in treated neonatal rats compared with their respective controls. Locomotor activities, such as crossing, grooming, rearing, and sniffing, were increased in ethanol-AGE-treated neonatal rats compared with controls. Reduced levels of intact pyramidal cells and cells with degenerative alterations appeared in the control rats. However, ethanol-AGE supplementation reduced degenerative alterations and hippocampal damage. Reduced cultured hippocampal neuron cell viability and increased apoptosis were noted in the control rats, whereas these impacts were significantly recovered following ethanol-AGE supplementation. Based on all these data, we concluded that the supplementation of AGE was very effective against ethanol-induced neurodegeneration in neonatal rats.

Environmentally triggered genomic plasticity and capsular polysaccharide formation are involved in increased ethanol and acetic acid tolerance in Kozakia baliensis NBRC 16680

BackgroundKozakia baliensis NBRC 16680 secretes a gum-cluster derived heteropolysaccharide and forms a surface pellicle composed of polysaccharides during static cultivation. Furthermore, this strain exhibits two colony types on agar plates; smooth wild-type (S) and rough mutant colonies (R). This switch is caused by a spontaneous transposon insertion into the gumD gene of the gum-cluster, resulting in a heteropolysaccharide secretion deficient, rough phenotype. To elucidate, whether this is a directed switch triggered by environmental factors, we checked the number of R and S colonies under different growth conditions including ethanol and acetic acid supplementation. Furthermore, we investigated the tolerance of R and S strains against ethanol and acetic acid in shaking and static growth experiments. To get new insights into the composition and function of the pellicle polysaccharide, the polE gene of the R strain was additionally deleted, as it was reported to be involved in pellicle formation in other acetic acid bacteria.ResultsThe number of R colonies was significantly increased upon growth on acetic acid and especially ethanol. The morphological change from K. baliensis NBRC 16680 S to R strain was accompanied by changes in the sugar contents of the produced pellicle EPS. The R:ΔpolE mutant strain was not able to form a regular pellicle anymore, but secreted an EPS into the medium, which exhibited a similar sugar monomer composition as the pellicle polysaccharide isolated from the R strain. The R strain had a markedly increased tolerance towards acetic acid and ethanol compared to the other NBRC 16680 strains (S, R:ΔpolE). A relatively high intrinsic acetic acid tolerance was also observable for K. baliensis DSM 14400T, which might indicate diverse adaptation mechanisms of different K. baliensis strains in altering natural habitats.ConclusionThe results suggest that the genetically triggered R phenotype formation is directly related to increased acetic acid and ethanol tolerance. The polE gene turned out to be involved in the formation of a cell-associated, capsular polysaccharide, which seems to be essential for increased ethanol/acetic tolerance in contrast to the secreted gum-cluster derived heteropolysaccharide. The genetic and morphological switch could represent an adaptive evolutionary step during the development of K. baliensis NBRC 16680 in course of changing environmental conditions.

Increasing Bioethanol Yield from Fermentation of Sweet Sorghum (Sorghum bicolor L. Moench) Sap by Mixed Culture Composed of Two Yeast Strains

Objectives: To evaluate the performance of the mixed culture of yeast strains to increase ethanol fermentation yield of sweet sorghum sap. Methods/Statistical Analysis:The yeast strains used were OUT7096, OUT7913, and OUT7921 while sweet sorghum saps utilized were KCS105 and FS501. Fermentation was performed in batch process and concentration of ethanol and sugars were determined by HPLC. Performance of each culture was indicated by ethanol yield and sugar conversion efficiency. The best culture was indicated by the best ethanol yield determined based on the one way ANOVA and Least Significant Difference test. Findings: The results showed that the composition of sugars and minerals in the two sweet sorghum saps are good for ethanol fermentation but supplementation with a bit of nitrogen and phosphorus may necessay for optimum yield. On the fermentation of KCS105 sap, all the three mixed culture especially OUT7913/ OUT7921 (1:1) provided higher performance than when used in single. On the fermentation of FS501 sap, the mixed culture of OUT7096/OU79013 (1:1) showed the highest performance among all of the cultures used, despite the data are not significant. The high value of SCE in both fermentation of KCS105 and FS501 saps suggested the oocurence of sugars other than sucrose, glucose and fructose. It was also found that mixed culture of OUT7913/OUT7921 (1:1) was more glucophilic than the mixed culture of OUT7096/OU79013 (1:1) . On the contrary, OUT7913/OUT7921 (1:1) culture was more fructophilic than the culture of OUT7913/OUT7921 (1:1) . Applications/Improvements: Mixed culture may improve ethanol yield of fermentation of substrate containing types of sugar like sweet sorghum sap.

Effects of end products on fermentation profiles in Clostridium carboxidivorans P7 for syngas fermentation

Clostridium carboxidivorans P7 is a strict anaerobic bacterium capable of converting syngas to biofuels. However, its fermentation profiles is poorly understood. Here, various end-products, including acetic acid, butyric acid, hexanoic acid, ethanol and butanol were supplemented to evaluate their effects on fermentation profiles in C. carboxidivorans at two temperatures. At 37°C, fatty acids addition likely led to more corresponding alcohols production. At 25°C, C2 and C4 fatty acids supplementation resulted in more corresponding higher fatty acids, while supplemented hexanoic acid increased yields of C2 and C4 fatty acids and hexanol. Supplementation of ethanol or butanol caused increased production of C2 and C4 acids at both temperatures; however, long-chain alcohols were still more likely produced at lower temperature. In conclusion, fermentation profiles of C. carboxidivorans can be changed in respond to pre-added end-products and carbon flow may be redirected to desired products by controlling culture conditions.

Oxidative damage and histopathological changes in lung of rat chronically exposed to nicotine alone or associated to ethanol

Smoking is the most important preventable risk factor of chronic obstructive pulmonary disease and lung cancer. This study was designed to investigate oxidative damage and histopathological changes in lung tissue of rats chronically exposed to nicotine alone or supplemented with ethanol. Twenty-four male Wistar rats divided into three groups were used for the study. The nicotine group received nicotine (2.5mg/kg/day); the nicotine-ethanol group was given simultaneously same dose of nicotine plus ethanol (0.2g/kg/day), while the control group was administered only normal saline (1ml/kg/day). The treatment was administered by subcutaneous injection once daily for a period of 18 weeks. Chronic nicotine administration alone or combined to ethanol caused a significant increase in malondialdehyde (MDA) level, superoxide dismutase (SOD) activity and catalase (CAT) activity in lung tissue compared to control rats suggesting an oxidative damage. However, these increases were mostly prominent in nicotine group. The histopathological examination of lung tissue of rats in both treated groups revealed many alterations in the pulmonary structures such as emphysema change (disappearance of the alveolar septa, increased irregularity and size of air sacs) and marked lymphocytic infiltration in perivascular and interstitial areas. However, the changes characterized in the nicotine group (pulmonary congestion, hemorrhage into alveoli and interstitial areas, edema) were more drastic than those observed in the nicotine-ethanol group, and they can be attributed to a significant degree of capillary endothelial permeability and microvascular leak. Conversely, the ethanol supplementation caused an appearance of fatty change and fibrosis in pulmonary tissue essentially due to a metabolism of ethanol. Finally, the lung damage illustrated in nicotine group was more severe than that observed in the nicotine-ethanol group. We conclude that the combined administration of nicotine and ethanol may moderate the effect of nicotine administered independently by counteractive interactions between these two drugs.

Assessment of bacterial acyltransferases for an efficient lipid production in metabolically engineered strains of E. coli

Microbially produced lipids like triacylglycerols or fatty acid ethyl esters are currently of great interest as fuel replacements or other industrially relevant compounds. They can even be produced by non-oleaginous microbes, like Escherichia coli, upon metabolic engineering. However, there is still much room for improvement regarding the yield for a competitive microbial production of lipids or biofuels. We genetically engineered E. coli by expressing fadD, fadR, pgpB, plsB and ‘tesA in combination with atfA from Acinetobacter baylyi. A total fatty acid contents of up to 16% (w/w) was obtained on complex media, corresponding to approximately 9% (w/w) triacylglycerols and representing the highest titers of fatty acids and triacylglycerols obtained in E. coli under comparable cultivation conditions, so far. To evaluate further possibilities for an optimization of lipid production, ten promising bacterial wax ester synthase/acyl-Coenzyme A:diacylglycerol acyltransferases were tested and compared. While highest triacylglycerol storage was achieved with AtfA, the mutated variant AtfA-G355I turned out to be most suitable for fatty acid ethyl ester biosynthesis and enabled an accumulation of approx. 500mg/L without external ethanol supplementation.

The Effect of Nicotine and its Interaction with Ethanol on Biochemical Parameters, Oxidative Damage and Histological Changes in the Rat’s Liver

In the present study we have evaluated the effect of chronic nicotine exposure and ethanol supplementation on biochemical parameters and oxidative stress in serum and liver of rats, respectively. We also performed histological analysis in liver. The nicotine-treated rats showed a weak effect on the activities of markers of liver function and a significant increase in the level of lactate dehydrogenase (LDH). The combined exposure significantly increased the level of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH). Moreover, the results showed that nicotine significantly increased lipid peroxidation, superoxide dismutase (SOD) activity and enhanced catalase (CAT) activity in liver. The combination of nicotine plus ethanol revealed the same effect in the level of lipid peroxidation and SOD activity, whereas the level of CAT activity was significantly raised. Histological studies showed morphological alterations in liver rat in both treated groups compared to control rats. Also, comparing the results of the nicotine-group to nicotine ethanol-group, we concluded that nicotine had prejudicial effects of less intensity than the association of nicotine and ethanol. These finding suggest that chronic nicotine administration provoked harmful effects to the liver; this hepatotoxicity can be enhanced by supplementation of ethanol and this association certainly increase the risk to develop liver diseases. Finally, the liver toxicity induced by nicotine alone or supplemented with ethanol was revealed by increasing both lipid peroxidation and oxidative stress in the liver, disturbance of indicators of liver function, enhance in the level of LDH and change of histological parameters.

Opuntia ficus indica f. inermis fruit juice alleviates ethanol-induced kidney injury in rats

The present work studied the protective effect of Opuntia ficus indica f. inermis fruit juice (OFIJ) against ethanol-induced renal dysfunction in rats. Chronic ethanol administration (3 g/kg b.w) during 12 weeks to Wistar rats, significantly (p < 0.01) increased the serum and kidney malondialdehyde (MDA), the serum contents of urea, creatinine, gamma glutamyl transferase (GGT) and the urinary glucose and protein levels. Whereas, copious ethanol intoxication significantly (p < 0.01) decreased the vitamin E and C levels in serum and kidney tissue as well as the glutathione content (GSH) and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and caused a severe renal histological injuries. Rivalry pre-treatment of ethanol-fed rats with OFIJ (20 and 40 ml/kg b.w., orally), interestingly attenuated the increases in serum and kidney MDA, and enhanced the antioxidant status of the rats by increasing the levels of SOD, CAT, (GSH-Px), GSH and vitamins. The biochemical findings positively corroborate with the amelioration of kidney histological morphology. The results suggest that the renal protective effect of Opuntia ficus indica f. inermis fruit juice (OFIJ) is by attenuating oxidative stress induced by copious ethanol supplement.

Improvement of ethanol production by ethanol-tolerant Saccharomyces cerevisiae UVNR56.

Ethanol tolerance is one of the important characteristics of ethanol-producing yeast. This study focused on the improvement of ethanol tolerance of Saccharomyces cerevisiae NR1 for enhancing ethanol production by random UV-C mutagenesis. One ethanol-tolerant mutant, UVNR56, displayed a significantly improved ethanol tolerance in the presence of 15% (v/v) ethanol and showed a considerably higher viability during ethanol fermentation from sugarcane molasses and sugarcane molasses with initial ethanol supplementation. A maximum ethanol concentration produced from molasses medium at 37°C by UVNR56 was 10.3% (v/v), productivity of 1.7 g/l/h and a theoretical yield of 98.7%, while the corresponding values for the wild-type were 8.6% (v/v), 1.4 g/l/h and 83.3%, respectively. In addition, during molasses fermentation under initial supplementation of 5% (v/v) ethanol, the maximum ethanol concentration and productivity of UVNR56 was 25.7% and 42.9% higher than the wild-type, respectively.

Endogenous Ethanol Affects Biopolyester Molecular Weight in Recombinant Escherichia coli

In biopolyester synthesis, polyhydroxyalkanoate (PHA) synthase (PhaC) catalyzes the polymerization of PHA in bacterial cells, followed by a chain transfer (CT) reaction in which the PHA polymer chain is transferred from PhaC to a CT agent. Accordingly, the frequency of CT reaction determines PHA molecular weight. Previous studies have shown that exogenous alcohols are effective CT agents. This study aimed to clarify the effect of endogenous ethanol as a CT agent for poly[(R)-3-hydroxybutyrate] [P(3HB)] synthesis in recombinant Escherichia coli, by comparing with that of exogenous ethanol. Ethanol supplementation to the culture medium reduced P(3HB) molecular weights by up to 56% due to ethanol-induced CT reaction. NMR analysis of P(3HB) polymers purified from the culture supplemented with (13)C-labeled ethanol showed the formation of a covalent bond between ethanol and P(3HB) chain at the carboxyl end. Cultivation without ethanol supplementation resulted in the reduction of P(3HB) molecular weight with increasing host-produced ethanol depending on culture aeration. On the other hand, production in recombinant BW25113(ΔadhE), an alcohol dehydrogenase deletion strain, resulted in a 77% increase in molecular weight. Analysis of five E. coli strains revealed that the estimated number of CT reactions was correlated with ethanol production. These results demonstrate that host-produced ethanol acts as an equally effective CT agent as exogenous ethanol, and the control of ethanol production is important to regulate the PHA molecular weight.

Impact of osmotic stress and ethanol inhibition in yeast cells on process oscillation associated with continuous very-high-gravity ethanol fermentation

BackgroundVHG fermentation is a promising process engineering strategy aiming at improving ethanol titer, and thus saving energy consumption for ethanol distillation and distillage treatment. However, sustained process oscillation was observed during continuous VHG ethanol fermentation, which significantly affected ethanol fermentation performance of the system.ResultsSustained process oscillation was investigated in continuous VHG ethanol fermentation, and stresses exerted on yeast cells by osmotic pressure from unfermented sugars and ethanol inhibition developed within the fermentation system were postulated to be major factors triggering this phenomenon. In this article, steady state was established for continuous ethanol fermentation with LG medium containing 120 g/L glucose, and then 160 g/L non-fermentable xylose was supplemented into the LG medium to simulate the osmotic stress on yeast cells under the VHG fermentation condition, but the fermentation process was still at steady state, indicating that the impact of osmotic stress on yeast cells was not the main reason for the process oscillation. However, when 30 g/L ethanol was supplemented into the LG medium to simulate the ethanol inhibition in yeast cells under the VHG fermentation condition, process oscillation was triggered, which was augmented with extended oscillation period and exaggerated oscillation amplitude as ethanol supplementation was increased to 50 g/L, but the process oscillation was gradually attenuated when the ethanol supplementations were stopped, and the steady state was restored. Furthermore, gas stripping was incorporated into the continuous VHG fermentation system to in situ remove ethanol produced by Saccharomyces cerevisiae, and the process oscillation was also attenuated, but restored after the gas stripping was interrupted.ConclusionsExperimental results indicated that ethanol inhibition rather than osmotic stress on yeast cells is one of the main factors triggering the process oscillation under the VHG fermentation condition, and in the meantime gas stripping was validated to be an effective strategy for attenuating the process oscillation.

Ethanol Promotes Arteriogenesis and Restores Perfusion to Chronically Ischemic Myocardium

Moderate alcohol consumption is known to be cardioprotective compared with either heavy drinking or complete abstinence. We assessed the hypothesis that ethanol supplementation would improve myocardial function in the setting of chronic ischemia. Sixteen male Yorkshire swine underwent placement of an ameroid constrictor into the left circumflex artery to induce chronic myocardial ischemia. Postoperatively, animals were supplemented with either 90 mL of ethanol (EtOH) daily (50%/V, EtOH) or 80 g of sucrose of equal caloric value (SUC), serving as controls. Seven weeks after ameroid placement, arteriolar density (1.74 ± 0.210% versus 3.11 ± 0.368% area of arterioles per low-powered field in sucrose (SUC) versus EtOH; P=0.004), myocardial perfusion (ratio of blood flow to the at-risk myocardium compared with the normal ventricle during demand pacing was 0.585 ± 0.107 versus 1.08 ± 0.138 for SUC versus EtOH; P=0.014), and microvascular reactivity were significantly increased in ethanol-treated animals compared with controls in the at-risk myocardium. Analysis of vascular endothelial growth factor and NOTCH pathway signaling suggested proneovascular and proliferative activity in the ischemic area. The average peak blood alcohol level in the treatment group was 40 ± 4 mg/dL, consistent with levels of moderate drinking in humans. Ethanol supplementation increased arteriolar density and significantly improved myocardial perfusion and endothelium-dependent vasorelaxation in chronically ischemic myocardium. These findings suggest that, at moderate doses, ethanol directly promotes vasculogenesis and improves microvascular function, resulting in significant improvements in myocardial perfusion in the setting of chronic ischemia.

Alcohol consumption improves insulin signaling in the myocardium

In a previous study, we demonstrated that swine with metabolic syndrome treated with alcohol had improved insulin signaling. We developed a follow-up study to evaluate the effects of alcohol on ischemic myocardium in animals without metabolic syndrome. Fourteen Yorkshire swine underwent placement of an ameroid constrictor to induce chronic myocardial ischemia. Postoperatively, one group was supplemented with ethanol (ETOH), and one group was supplemented with sucrose (SUC) daily to normalize caloric intake. After 7 weeks, all animals underwent dextrose challenge and harvest of nonischemic and ischemic myocardium. Tissues were analyzed for protein expression and histologic analysis. There was no difference in body mass index, serum glucose or insulin levels. However, ethanol supplementation up-regulated phosphoinostitide 3-kinase, phosphorylated protein kinase B, protein kinase B, and phosphorylated Forkhead Box 01 expression, which may promote insulin signaling, and down-regulated inhibitors of insulin signaling pIRS1 and pIRS2. There was no difference in intramyocardial glycogen but there was increased GLUT4 expression in the ETOH group, which may promote glucose use. Despite similar serum glucose and insulin levels, alcohol consumption up-regulates the insulin signaling pathway in the absence of metabolic syndrome in both nonischemic and chronically ischemic myocardium. These results suggest that alcohol selectively up-regulates the insulin signaling pathway despite normoglycemia.

Synthesis of medium chain length fatty acid ethyl esters in engineered Escherichia coli using endogenously produced medium chain fatty acids

Microbial biosynthesis of fatty acid-derived biofuels from renewable carbon sources has attracted significant attention in recent years. Free fatty acids (FFAs) can be used as precursors for the production of micro-diesel. The expression of codon optimized two plants (Umbellularia californica and Cinnamomum camphora) medium-chain acyl–acyl carrier protein (ACP) thioesterase genes (ucFatB and ccFatB) in Escherichia coli resulted in a very high level of extractable medium-chain-specific hydrolytic activity and caused large accumulation of medium-chain free fatty acids. By heterologous co-expression of acyl-coenzyme A:diacylglycerol acyltransferase from Acinetobacter baylyi ADP1, specific plant thioesterases in E. coli, with supplementation of exogenous ethanol, resulted in drastic changes in fatty acid ethyl esters (FAEEs) composition ranging from 12:0 to 18:1. Through an optimized microbial shake-flask fermentation of two modified E. coli strains, yielded FFAs and FAEEs in the concentration of approximately 500mgL−1/250mgL−1 and 2.01mgg−1/1.99mgg−1, respectively. The optimal ethanol level for FAEEs yield in the two recombinant strains was reached at the 3% ethanol concentration, which was about 5.4-fold and 1.93-fold higher than that of 1% ethanol concentration.

Alcohol treatment enhancesStaphylococcus aureusbiofilm development

Staphylococcus aureus forms pathogenic biofilms. Previous studies have indicated that ethanol supplementation during S.aureus biofilm formation results in increased biofilm formation and changes in gene expression. However, the impact of alcohols on preformed S.aureus biofilms has not been studied. In this study, we formed S.aureus biofilms on PVC plastic plates and then treated these preformed biofilms with five different alcohols. We observed that alcohol treatment of preformed S.aureus biofilms led to significant increases in biofilm levels after 24h of treatment. Many bacteria within these biofilms were found to be alive and metabolically active. Alcohol treatment also resulted in increased transcription of the biofilm-promoting genes icaA and icaD, as well as several antibiotic resistance genes. These results demonstrate that treatment of S.aureus preformed biofilms with alcohols enhances biofilm levels if maintained for extended periods. Thus, alcohols might be of limited usefulness for the eradication of preformed S.aureus biofilms.