Ethanol Treatment Research Articles

Effect of microwave and enzymatic pretreatment and type of solvent on kinetics of ultrasound assisted extraction of bioactive compounds from ripe papaya peel

AbstractThe study was aimed to investigate the effect of pretreatment by microwave and enzyme and type of solvent on kinetics of ultrasound‐assisted extraction (UAE) of bioactive compounds and antioxidant activity from papaya peels. The enzyme used was viscozyme and the solvents used were water, ethanol, and their combination (1:1). The effect of combination of pretreatments was also studied. The data from the experiments were fitted to second order kinetic model and Langmuir model. The second order rate constant (k) varied from 0.0005 to 0.0134 for total phenolic content (TPC), 0.0016–0.0080 for total flavonoid content, and 0.0004–0.0228 for antioxidant activity (2,2‐diphenyl‐1‐picrylhydrazyl‐radical scavenging activity). The comparative kinetic assessment illustrated that the progression of the extraction for TPC was higher in enzyme‐water (E‐W) treatment (98.91 mg Gallic acid equivalent (GAE)/100 g), followed by microwave‐enzyme water (ME‐W) treatment (93.98 mg GAE/100 g) while microwave‐water:ethanol (M‐WEt) treatment gave the lowest yield (42.56 mg GAE/100 g), respectively. The present study also investigated influences of ultrasonication on extraction kinetics and their yield. Ultrasound was clearly influencing the yield (over 72.95% increase in yield of polyphenols). Antioxidant activity of the extracts confirmed that the UAE extraction is suitable for preparing antioxidant‐rich plant extracts. Ultrasound and pretreatment before extraction could provide many advantages like increased yield and quality, energy efficiency, faster extraction, and low solvent consumption.Practical ApplicationsPapaya peels are waste products generated during processing of papaya and are valuable sources of polyphenols and natural antioxidants such as caffeic acid, rutin, ferulic acid, p‐coumaric acid, and many more bioactive compounds that have significant biological activity. These bioactive compounds of papaya peel can be extracted and utilized in food and pharmaceutical industry for development of functional food or nutraceuticals. Ultrasound extraction is regarded as a green technology as it is environment friendly and energy‐efficient process. When ultrasound technology is combined with microwave or enzymatic treatment, the efficiency of such process may be further improved. The present study is therefore useful as it can provide a solution for utilization of papaya peel for extraction of valuable compounds using green technology.

Maternal Ethanol Exposure-Induced Cardiac Fibrosis is Associated with Changes in TGF-β and SIRT1/FOXO3a Signaling in Male Rat Offspring: A Three-Month Follow-up Study.

Ethanol exposure during pregnancy induces cardiac fibrosis inthe fetalheart. However, the mechanisms by which consumption of ethanol induces fibrotic changes are not known. Pregnant rats were received ethanol 4.5g/kg BW once per day from the 7th day of pregnancy (GD7) throughout lactation. Our findings demonstrated that, area of fibrosis increased in cardiac tissue in the pups on both postnatal day twenty one (PN21) and postnatal day ninety (PN90) after prenatal and early postnatal period ethanol treatment compared with the controls. It was accompanied by a decline in the expression of SIRT1 protein along with the elevation of FOXO3a and TGF-β protein expressions which weredetermined by western blot. Overall, our data reveal that prenatal alcohol usage increase in fibrotic regions in the pup hearts possibly by regulating TGF-β, FOXO3a and SIRT1 protein levels. These are potential therapeutic molecular targets that can be modulated to protect heart against maternal ethanol exposure.

Anchoring and reactivation of single-site Co–porphyrin over TiO2 for the efficient photocatalytic CO2 reduction

Porphyrin and its derivatives have been often used for electrochemical and photocatalysis combined with first-row transition metals for sustainable applications. Cobalt–tetrakis(4-carboxyphenyl)porphyrin (Co–TCPP) was covalently anchored onto TiO2. Under CO2, H2, and UV–visible light irradiation, Co–TCPP anchored on TiO2 produced CO at a rate of 2.5 μmol h−1 (using 100 mg of catalyst and UV–visible light of 118 mW cm−2) when the Co–TCPP content was 2.5 wt %; however, the CO formation rate dropped to 0.3 μmol h−1 (using 100 mg of catalyst and the same intensity of light). The photoactivity drop to 12 % was solved by removing the hydroxy ligands coordinated to the central Co ions through ethanol treatment for 5 min of the Co–TCPP–TiO2 photocatalysts. This resulted in an increased CO photoformation rate to 7.3 μmol h−1 (using 100 mg of catalyst and the same intensity of light) because the dispersion of the J- and/or H-aggregates to the isolated Co–TCPP unit proceeded more with ethanol treatment and under UV–visible light. Conversely, to use ethanol as reductant was not effective because it reduced and deactivated Co–TCPP.

Optimal Solvents for Interfacial Solution Engineering of Perovskite Solar Cells

Interfacial engineering is extensively used to reduce the interfacial loss caused by surface recombination, improve the crystallinity of the active absorption layer, and enhance the long‐term stability of perovskite solar cells (PSCs). Solution processing techniques, such as spin coating and dip coating, are commonly used to deposit the interfacial layer because of their cost‐effectiveness and simplicity. Although determining suitable solutes for use in these processes is important, selecting appropriate solvents is also crucial. Herein, commonly used solvents are investigated to determine optimal solvents for solution processing by categorizing them into nonpolar and polar groups. The results suggest that the efficiency of the PSCs can be increased by simple solvent treatment. In particular, the efficiencies of systems subjected to hexane (nonpolar) and ethanol (polar) treatment are significantly improved (17.31% and 17.44%, respectively) compared with that of a control device (16.24%). Herein, the effects of pure solvents on the SnO2–perovskite interface are confirmed and an important direction for investigations that adopt solution processing to improve the efficiency of PSCs, such as research on interlayers and self‐assembled monolayers, is suggested.

γ-Glutamylcysteine alleviates ethanol-induced hepatotoxicity via suppressing oxidative stress, apoptosis, and inflammation.

Alcohol abuse is a major cause of alcoholic liver disease (ALD) and can result in fibrosis and cirrhosis. γ-glutamylcysteine (γ-GC) is a precursor of glutathione (GSH) with antioxidant and anti-inflammatory properties. Our research aimed to explore the protective impact of γ-GC on ALD and its potential mechanisms of efficiency in vitro and in vivo. L02 cells were pretreated with γ-GC (20, 40, and 80 μM) for 2h and exposed to ethanol for 24 h. Cell viability, apoptosis, oxidative stress, and inflammatory levels were measured. The expression of protein cleaved caspase-3 and cleaved PARP and flow cytometry results indicated that γ-GC decreases apoptosis on L02 cells after ethanol treatment. Moreover, γ-GC also attenuated oxidative stress and mitochondrial damage in hepatocytes caused by ethanol via increasing cellular GSH, SOD activity, and mitochondrial membrane potential. In vivo experiments, γ-GC effectively reduced the AST, ALT, and TG levels in mice. The inflammation of ALD was alleviated by γ-GC both in vivo and in vitro. Additionally, histopathological examination demonstrated that γ-GC treatment lessened lipid droplet formation and inflammatory damage. In conclusion, these results showed that γ-GC has anti-inflammatory and anti-apoptotic effects on ALD because it could help hepatocytes maintain sufficient GSH levels to combat the excess reactive oxygen species (ROS) generated during ethanol metabolism. PRACTICAL APPLICATIONS: Alcohol intake is the fifth highest risk factor for premature death and disability among all risk variables. However, few medicines are both safe and effective for the treatment of ALD. As a direct precursor of GSH, γ-GC has a broad variety of potential antioxidant and anti-inflammatory applications for the treatment of numerous medical conditions. In conclusion, these results showed that γ-GC could protect cells from ALD via suppressing oxidative stress, alleviating inflammation, and preventing apoptosis.

Kumagawa and Soxhlet Solvent Fractionation of Lignin: The Impact on the Chemical Structure.

We investigated the effects of solvent fractionation on the chemical structures of two commercial technical lignins. We compared the effect of Soxhlet and Kumagawa extraction. The aim of this work was to compare the impact of the methods and of the solvents on lignin characteristics. Our investigation confirmed the potentialities of fractionation techniques in refining lignin properties and narrowing the molecular weight distribution. Furthermore, our study revealed that the Kumagawa process enhances the capacity of oxygenated solvents (ethanol and tetrahydrofuran) to extract lignin that contains oxidized groups and is characterized by higher average molecular weights. Furthermore, the use of tetrahydrofuran after ethanol treatment enabled the isolation of lignin with a higher ratio between carbonyl and other oxidized groups. This result was confirmed by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), 13C NMR and two-dimensional (2D) NMR spectroscopies, gel permeation chromatography (GPC), and analytical pyrolysis-gas chromatography–mass spectrometry (Py-GC–MS) analysis. Ultraviolet–visible (UV–vis) spectra evidenced the enrichment in the most conjugated species observed in the extracted fractions. Elemental analyses pointed at the cleavage of C-heteroatom bonds enhanced by the Kumagawa extraction.

Effects of Ethanol on Expression of Coding and Noncoding RNAs in Murine Neuroblastoma Neuro2a Cells.

Excessive use of alcohol can induce neurobiological and neuropathological alterations in the brain, including the hippocampus and forebrain, through changes in neurotransmitter systems, hormonal systems, and neuroimmune processes. We aimed to investigate the effects of ethanol on the expression of coding and noncoding RNAs in a brain-derived cell line exposed to ethanol. After exposing Neuro2a cells, a neuroblastoma cell line, to ethanol for 24 and 72 h, we observed cell proliferation and analyzed up- and downregulated mRNAs and long noncoding RNAs (lncRNAs) using total RNA-Seq technology. We validated the differential expression of some mRNAs and lncRNAs by RT-qPCR and analyzed the expression of Cebpd and Rnu3a through knock-down of Cebpd. Cell proliferation was significantly reduced in cells exposed to 100 mM ethanol for 72 h, with 1773 transcripts up- or downregulated by greater than three-fold in ethanol-treated cells compared to controls. Of these, 514 were identified as lncRNAs. Differentially expressed mRNAs and lncRNAs were mainly observed in cells exposed to ethanol for 72 h, in which Atm and Cnr1 decreased, but Trib3, Cebpd, and Spdef increased. On the other hand, lncRNAs Kcnq1ot1, Tug1, and Xist were changed by ethanol, and Rnu3a in particular was greatly increased by chronic ethanol treatment through inhibition of Cebpd. Our results increase the understanding of cellular and molecular mechanisms related to coding and noncoding RNAs in an in vitro model of acute and chronic exposure to ethanol.

Surface Treatment With Hydrophobic Coating Reagents (Organosilanes) Strongly Reduces the Bioactivity of Synthetic Amorphous Silica in vitro.

Synthetic amorphous silica (SAS) is industrially relevant material whose bioactivity in vitro is strongly diminished, for example, by protein binding to the particle surface. Here, we investigated the in vitro bioactivity of fourteen SAS (pyrogenic, precipitated, or colloidal), nine of which were surface-treated with organosilanes, using alveolar macrophages as a highly sensitive test system. Dispersion of the hydrophobic SAS required pre-wetting with ethanol and extensive ultrasonic treatment in the presence of 0.05% BSA (Protocol 1). Hydrophilic SAS was suspended by moderate ultrasonic treatment (Protocol 2) and also by Protocol 1. The suspensions were administered to NR8383 alveolar macrophages under serum-free conditions for 16 h, and the release of LDH, GLU, H2O2, and TNFα was measured in cell culture supernatants. While seven surface-treated hydrophobic SAS exhibited virtually no bioactivity, two materials (AEROSIL® R 504 and AEROSIL® R 816) had minimal effects on NR8383 cells. In contrast, non-treated SAS elicited considerable increases in LDH, GLU, and TNFα, while the release of H2O2 was low except for CAB-O-SIL® S17D Fumed Silica. Dispersing hydrophilic SAS with Protocol 1 gradually reduced the bioactivity but did not abolish it. The results show that hydrophobic coating reagents, which bind covalently to the SAS surface, abrogate the bioactivity of SAS even under serum-free in vitro conditions. The results may have implications for the hazard assessment of hydrophobic surface-treated SAS in the lung.

MyD88 in hepatic stellate cells promotes the development of alcoholic fatty liver via the AKT pathway.

Myeloid differentiation primary response gene 88 (MyD88), an adaptor protein in the Toll-like receptors (TLRs) signalling pathway, is expressed in various liver cells including hepatocytes, Kupffer cells and hepatic stellate cells (HSCs). And yet, the functional role of MyD88 in HSCs is poorly elucidated in alcoholic fatty liver (AFL). Here, to study the functional role of MyD88 in HSCs and the molecular mechanism related to the development of AFL, chronic-binge ethanol mouse models were established in mice with specific MyD88 knockout in quiescent (MyD88GFAP-KO) and activated HSCs (MyD88SMA-KO), respectively. Our results clearly showed an elevated expression of MyD88 in liver tissues of ethanol treated mouse model which harbours the wild type. Intriguingly, ethanol treatment profoundly inhibited inflammation in both MyD88GFAP-KO and MyD88SMA-KO mice, but the suppression of lipogenesis was only observed in MyD88GFAP-KO mice. Molecularly, our study indicated that MyD88 induced osteopontin (OPN) secretion in HSCs, which consequently resulted in activation of AKT signalling pathway and accumulation of fat in hepatocytes. Additionally, our data also suggested that OPN promoted inflammation by activating p-STAT1. Thus, targeting MyD88 may be a potentially represent a promising strategy for the prevention and treatment of AFL. KEY MESSAGES: The expression of MyD88 in HSCs was significantly increased in ethanol-induced liver tissues of wild-type mice. MyD88 deficiency in quiescent HSCs inhibited inflammation and lipogenesis under the ethanol feeding condition. MyD88 deficiency in activated HSCs only inhibited inflammation under the ethanol feeding condition. MyD88 promoted the OPN secretion of HSCs, which further activated the AKT signalling pathway of hepatocytes and upregulated lipogenic gene expression to promote fat accumulation. OPN also promotes inflammation by activating p-STAT1.

Effects of Ethanol Treatment on Storage Quality and Antioxidant System of Postharvest Papaya.

Papaya is the fourth most favored tropical fruit in the global market; it has rich nutrition and can be used for medicine and food processing. However, it will soften and mature in a short time after harvest, resulting in a lot of economic losses. In this study, papaya fruits were soaked in 0, 12.5, 25, 50, and 100 ml/L ethanol solutions for 2 h and stored at 25°C for 14 days, by which we explored the effects of ethanol treatment in papaya after harvest. At an optimal concentration of ethanol treatment, color changing of the papaya fruits was delayed for 6 days, and decay incidence and average firmness of the fruits were shown as 20% and 27.7 N, respectively. Moreover, the effect of ethanol treatment on antioxidant systems in the papaya fruits was explored. It was observed that ethanol treatment contributed to diminish the development of malondialdehyde (MDA), ethylene, and superoxide anions. Furthermore, the activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) were promoted than those of control group, while the activities of peroxidase (POD), phenylalanine ammonia-lyase (PAL), and polyphenol oxidase (PPO) were brought down. In addition, the principal component analysis (PCA) showed that PAL, ethylene, and superoxide anions were the main contributors for the maturity and senescence of postharvest papaya. In this experiment, ethanol treatment had the potential of delaying the ripening and maintaining the storage quality of papaya fruits.

Lethal effect of ethanol on Solenopsis invicta Buren and economic fish

AbstractThe red imported fire ant (RIFA), Solenopsis invicta Buren, is an invasive pest species worldwide in many countries and regions. At present, its control mainly depends on synthetic insecticides. Although synthetic insecticides have the advantages of efficacy and convenience, their widespread use has a negative impact on the diversity of local arthropods. Ethanol is widely used as a disinfectant and has been consumed by people, but its insecticidal effect has not yet been evaluated. Here, the control effect of ethanol on the RIFA and its toxicity on four fish species were evaluated. The results exhibited that mortalities of all tested adult castes reached 100% after soaking in 70% v/v ethanol solution for one hour. All reproductive and worker pupae survived at 60% v/v ethanol treatment. In the soaking plus fumigation experiment, all adult castes and brood except female pupae were sensitive to 30% v/v ethanol solution. Conversely, female pupae showed the highest resistance to ethanol. More than 60% v/v ethanol concentration was needed to cause its 100% mortality. In the field experiment, 25% v/v and 50% v/v ethanol solution resulted in 70% and 80% reduction of mound 14 d after drench treatment, respectively. The ethanol vapour in the nest soil was higher than 80 ppm after 72 h of mound drench in the field with 25% v/v ethanol solution. Moreover, 30% v/v ethanol solution did not adversely affect the survival of four fish species exposed to the solution for a short time. This study suggests that mound drench can be alternatively employed with ethanol as a green organic solvent to control RIFA.

Synthesis, In Silico and Pharmacological Evaluation of New Thiazolidine-4-Carboxylic Acid Derivatives Against Ethanol-Induced Neurodegeneration and Memory Impairment

IntroductionSeveral studies revealed that alcohol utilization impairs memory in adults; however, the underlying mechanism is still unclear. The production of inflammatory markers and reactive oxygen species (ROS) plays a major role in neurodegeneration, which leads to memory impairment. Therefore, targeting neuroinflammation and oxidative distress could be a useful strategy for abrogating the hallmarks of ethanol-induced neurodegeneration. Moreover, several studies have demonstrated multiple biological activities of thiazolidine derivatives including neuroprotection.MethodsIn the current study, we synthesized ten (10) new thiazolidine-4-carboxylic acid derivatives (P1-P10), characterized their synthetic properties using proton nuclear magnetic resonance (1H-NMR) and carbon-13 NMR, and further investigated the neuroprotective potential of these compounds in an ethanol-induced neuroinflammation model.ResultsOur results suggested altered levels of antioxidant enzymes associated with an elevated level of tumor necrosis factor-alpha (TNF-α), nuclear factor-κB (p-NF-κB), pyrin domain-containing protein 3 (NLRP3), and cyclooxygenase-2 (COX-2) in ethanol-treated animals. Ethanol treatment also led to memory impairment in rats, as assessed by behavioral tests. To further support our notion, we performed molecular docking studies, and all synthetic compounds exhibited a good binding affinity with a fair bond formation with selected targets (NF-κB, TLR4, NLRP3, and COX-2).DiscussionOverall, our results revealed that these derivatives may be beneficial in reducing neuroinflammation by acting on different stages of inflammation. Moreover, P8 and P9 treatment attenuated the neuroinflammation, oxidative stress, and memory impairment caused by ethanol.

Rapid anticalcification treatment for glutaraldehyde-fixed autologous tissue in cardiovascular surgery

BackgroundGlutaraldehyde (GA)-fixed autologous tissues, including the pericardium, are widely used as patches and valve substitutes in cardiovascular surgery. However, GA treatment causes tissue calcification. No rapid anticalcification method has been established for use during surgery. Here, we aimed to establish a rapid anticalcification method using ethanol, as has already been demonstrated for bioprosthetic valves.MethodsThoracic aorta tissues were first fixed with GA for 3 min and then treated with ethanol for 0 (group 2), 10 (group 3), 20 (group 4), and 30 (group 5) min; untreated tissues (group 1) served as the control. The treated tissues were subdermally implanted into 3-week-old male Wistar rats and kept in place for 28 days. The calcification in each explant was semiquantitatively evaluated by annotating and measuring the area using virtual slides, and the data obtained were statistically analyzed.ResultsSemiquantitative analysis revealed that calcification of the implants from the untreated group (group 1; P = 0.0014) and groups 4 (P = 0.0014) and 5 (P = 0.0031) was significantly lower than that of implants from group 2. Moreover, implants from group 3 showed a tendency toward decreased calcification, although it was not significant (P = 0.0503).ConclusionsA rapid ethanol treatment prevents calcification of GA-fixed tissues in a rat model of subdermal implantation. This method may facilitate effective and rapid anticalcification of autologous tissues for use during cardiovascular surgery.

Study on Soda–Ethanol Delignification of Pine Sawdust for a Biorefinery

The soda–ethanol process was conceived as a sulfur-free pulping process, which may also be an alternative to conventional alkaline pulping, such as kraft or soda–AQ in the biorefinery context. An in-depth study using two experimental designs was conducted to establish the viability of soda–ethanol delignification of pine sawdust. At first, a simple factorial design involving the ethanol–water ratio (ethanol:water) and the alkaline load (AL, % over dry wood, odw) was applied to define the levels of these variables and their eventual interaction. Then, a 32 experimental design was performed to evaluate the ability of the process concerning the pulping of pine sawdust. The tested conditions were carefully selected to screen a broad range of cooking times (60, 100, and 140 min) and alkaline loads (19.0, 23.3, and 27.6 %odw) to obtain pulps with different extents of delignification (residual lignin contents). Finally, the kraft, soda–AQ, and soda–ethanol treatments were compared. Soda–ethanol pulping was shown to be a suitable delignification stage for a biorefinery scheme of Pinus elliottii and Pinus taeda sawdust. It has many advantages over traditional processes regarding its environmental impact, harmless chemicals, and selectivity. The tested conditions were similar to those frequently used in conventional pulping at an industrial scale, suggesting the technical feasibility of the soda–ethanol process for pine sawdust processing.

Chronic ethanol consumption plus an atherogenic diet cause metabolic steatohepatitis with advanced liver fibrosis in apolipoprotein E/low-density lipoprotein receptor double-knockout mice.

Nonalcoholic steatohepatitis is the inflammatory subtype of nonalcoholic fatty liver disease with a high risk of progression to liver fibrosis. We investigated metabolic steatohepatitis with advanced liver fibrosis in apolipoprotein E/low-density lipoprotein receptor double-knockout (AL) mice fed a co-diet of ethanol with a low-carbohydrate-high-protein-high-fat atherogenic diet (AD) for 16weeks. We also examined the underlying mechanisms, especially hepatic sympathetic activation, involved in the effects. We maintained 12-week-old male AL mice on AD and a standard chow diet (SCD) with or without ethanol treatment for 16weeks. Age-matched male C57BL/6J mice on SCD without ethanol treatment served as controls. We conducted blood biochemical, histopathological, and fluorescence immunohistochemical, and reverse transcriptase polymerase chain reaction studies. AL mice showed significant hyperlipidemia. AD induced increased body weight, hepatic steatosis, and hepatic damage; ethanol and the AD co-diet resulted in hepatic sympathetic activation accompanied by hepatic steatosis, lobular inflammation, bridging fibrosis, and hepatic damage. Hepatic Kupffer cells (KCs) and hepatic stellate cells (HSCs), which showed sympathetic activation, produced 4.4- to 9.4-fold more inflammatory factors (KC and KC-derived tumor necrosis factor-α, and chemokine [C-C motif] ligand 2) and 2.0- to 32.0-fold more fibrosis factors (HSC and HSC-derived transforming growth factor β1 and collagen 1a1); all p<0.05 vs. controls. We created a model of metabolic steatohepatitis with advanced liver fibrosis from coexisting hyperlipidemia and hepatic sympathetic activation in AL mice on a co-diet of ethanol and AD. KCs and HSCs became the cellular targets of hepatic sympathetic activation, which could play a role in the initiation and progression of metabolic steatohepatitis with advanced liver fibrosis.

Beer ethanol and iso-α-acid level affect microbial community establishment and beer chemistry throughout wood maturation of beer

Sour beers produced by barrel-aging of conventionally fermented beers are becoming increasingly popular. However, as the intricate interactions between the wood, the microbes and the beer are still unclear, wood maturation often leads to inconsistent end products with undesired sensory properties. Previous research on industrial barrel-aging of beer suggests that beer parameters like the ethanol content and bitterness play an important role in the microbial community composition and beer chemistry, but their exact impact still remains to be investigated. In this study, an experimentally tractable lab-scale system based on an in-vitro community of four key bacteria (Acetobacter malorum, Gluconobacter oxydans, Lactobacillus brevis and Pediococcus damnosus) and four key yeasts (Brettanomyces bruxellensis, Candida friedrichii, Pichia membranifaciens and Saccharomyces cerevisiae) that are consistently associated with barrel-aging of beer, was used to test the hypotheses that beer ethanol and bitterness impact microbial community composition and beer chemistry. Experiments were performed using different levels of ethanol (5.2 v/v%, 8 v/v% and 11 v/v%) and bitterness (13 ppm, 35 ppm and 170 ppm iso-α-acids), and beers were matured for 60 days. Samples were taken after 0, 10, 20, 30 and 60 days to monitor population densities and beer chemistry. Results revealed that all treatments and the maturation time significantly affected the microbial community composition and beer chemistry. More specifically, the ethanol treatments obstructed growth of L. brevis and G. oxydans and delayed fungal growth. The iso-α-acid treatments hindered growth of L. brevis and stimulated growth of P. membranifaciens, while the other strains remained unaffected. Beer chemistry was found to be affected by higher ethanol levels, which led to an increased extraction of wood-derived compounds. Furthermore, the distinct microbial communities also induced changes in the chemical composition of the beer samples, leading to concentration differences in beer- and wood-derived compounds like 4-ethyl guaiacol, 4-ethyl phenol, cis-oak lactone, vanillin, furfural and 5-hydroxymethyl furfural. Altogether, our results indicate that wood-aging of beer is affected by biotic and abiotic parameters, influencing the quality of the final product. Additionally, this work provides a new, cost-effective approach to study the production of barrel-aged beers based on a simplified microbial community model.

A Comparative Genome-Wide Transcriptome Analysis of Glucocorticoid Responder and Non-Responder Primary Human Trabecular Meshwork Cells.

Aim: To investigate genes and pathways involved in differential glucocorticoid (GC) responsiveness in human trabecular meshwork (HTM) cells using RNA sequencing. Methods: Using paired human donor eyes, human organ-cultured anterior segment (HOCAS) was established in one eye to characterize GC responsiveness based on intra ocular pressure (IOP) change and, in the other eye, primary HTM cell culture was established. For RNA sequencing, total RNA extracted from GC-responder (GC-R) and non-responder (GC-NR) cells after dexamethasone (DEX) or ethanol (ETH) treatment for 7d was used. Differentially expressed genes (DEGs) were compared among five groups and validated. Results: In total, 616 and 216 genes were identified as significantly dysregulated in Group #1 and #2 (#1: ETH vs. DEX-treated GC-R; #2: ETH vs. DEX-treated GC-NR), respectively. Around 80 genes were commonly dysregulated in Group #3 (overlapping DEGs between #1 and #2), whereas 536 and 136 genes were uniquely expressed in GC-R (#4) and GC-NR HTM (#5) cells, respectively. Pathway analysis revealed that WNT signaling, drug metabolism cytochrome p450, cell adhesion, TGF-β signaling, and MAPK signaling were associated with GC responsiveness. Conclusion: This is the first study reporting distinct gene signatures and their associated pathways for GC-R and GC-NR HTM cells. WNT and MAPK signaling are potential therapeutic targets for the management of GC-induced glaucoma.

Time-Dependent Alterations in Liver Oxidative Stress due to Ethanol and Acetaldehyde

Binge drinking is a major public health issue and ethanol-related liver insult may play a major role in the pathology of alcoholic liver disease. However, the degree of oxidative stress, cell death and contribution of acetaldehyde to liver damage over a 24-h period has yet to be determined. Herein, we aimed to investigate the effect of acute alcohol and elevated acetaldehyde levels on hepatic oxidative damage, apoptosis, and antioxidant enzyme activity over a 24-h period. Male Wistar rats were divided into four groups and animals were pre-injected (intraperitonially [i.p.]) with either saline (0.15 mol/L) or cyanamide (5-mmol/kg body weight), followed by either saline (0.15 mol/L) or ethanol (75-mmol/kg bodyweight). After 2.5, 6 and 24 h, hepatic cytosolic and mitochondrial fractions were analysed for indices of oxidative stress. At 2.5 h, cytosolic glutathione and malondialdehyde levels were significantly reduced and increased, respectively, with alcohol treatment. Caspase-3 activity and cytochrome c levels were increased with alcohol treatment at 24 h. The combination of cyanamide and alcohol treatment at 24 h led to a significant increase in serum alanine aminotransferase levels, and reduced albumin and total protein levels. Furthermore, glutathione peroxidase activity and glutathione reductase activity were significantly decreased and increased, respectively. Finally, superoxide dismutase activity was decreased in cytosol and increased in the mitochondria after cyanamide and ethanol treatment, respectively. This study indicates a complex differential effect of alcohol and acetaldehyde, whereby alcohol toxicity in the mitochondria takes place throughout the 24-h period, but raised acetaldehyde has a further detrimental effect on liver function.

Characterizing the Real‐Time Endogenous Methane Release in a Rat for 26‐days: Effect of Diets, Fasting, and Ethanol Consumption Across Different Time Scales

IntroductionMethane is a byproduct of methanogenic archaea that oxidize H2 gas produced by other microflora in the gut. This methane gas is readily absorbed into the into systemic circulation and excreted in the breath down its concentration gradient (similar to CO2) or may be acutely expelled as flatus. While ruminant animals can generate over 100 liters of methane per day, technological limitations have precluded detailed measurements of methane production and release in humans and other non‐ruminants.MethodsWe paired a Promethion metabolic measurement system with a high‐precision (&lt;1ppb) methane analyzer capable of real‐time methane measurements in an individual mouse or rat. We then subjected a rat to multiple diet switches, 48h of fasting, and ethanol treatments and continuously measured food/water intake, metabolic rate, and methane release over 26 days.ResultsVCO2 responses were unaffected by diet changes and only decreased slightly during the fasting treatment. Methane release was diminished by some diet treatments and could be restored by others. Ethanol had no observable effects. We discovered that methane production generally followed a circadian cycle whereby total rates were ~50% higher and large flatus events were more frequent during the scotophase. Methane release during flatus varied by &gt;1,000‐fold ranging from as little as 0.1 up to 60μl/min. Integrating the continuous data stream (at 1 Hz) we confirmed that the vast majority of methane release occurred in the breath and not in the flatus as is often repeated in the literature. Finally, we provide evidence that the constitutive rate of methane release in the breath is positively correlated with rates of CO2 release, the latter of which is indicative of overall activity levels of the rat.ConclusionGiven the growing number of molecular‐based studies investigating how gut microbiota can alter physiological function at higher levels of biological organization the approach of using real‐time methane measurements promises to provide a powerful new tool to establish causal linkages between microbiome dynamics and whole‐animal responses.

6-Methoxyflavone and Donepezil Behavioral Plus Neurochemical Correlates in Reversing Chronic Ethanol and Withdrawal Induced Cognitive Impairment.

PurposeChronic ethanol exposure causes neurotoxicity and long-term learning and memory impairment along with hippocampal and frontal cortical dysfunction. Flavonoids possess antioxidant and anti-inflammatory properties believed to be contributory factors in reversing cognitive decline. 6-Methoxyflavone (6-MOF), a flavonoid occurring naturally in medicinal plants, has been reported to instigate neuroprotection by reversing cisplatin-induced hyperalgesia and allodynia. Consequently, this study was designed to investigate 6-MOF activity in models of chronic ethanol-induced cognitive impairment along with neurochemical correlates.MethodsMice were given ethanol orally (2.0 g/kg daily) for 24 days plus either saline, 6-MOF (25–75mg/kg) or donepezil (4mg/kg) and then ethanol was withdrawn for the next 6 days. Animals were subsequently assessed for their cognitive performance in several models on days 1, 12, and 24, during abstinence (Day-26) and on the 7th day of the washout period. Following behavioral assessment, post-mortem dopamine, noradrenaline and vitamin C concentrations were quantified in the frontal cortex, hippocampus and striatum, using HPLC with UV detection.ResultsChronic ethanol treatment suppressed locomotor activity and impaired cognitive tasks, which included novel object recognition, performance in the Morris water maze as well as the Y-maze, socialization and nest-building behavior throughout the protocol and during withdrawal. These behavioral deficits were at least partially restored by the co-administration of 6-MOF or donepezil with ethanol as were ethanol-induced deficits in frontal cortical and hippocampal dopamine plus noradrenaline, together with striatal dopamine. 6-MOF co-administration with ethanol also modestly restored striatal vitamin C levels.ConclusionIt is postulated that, apart from donepezil, 6-MOF may be useful not only in the treatment of ethanol withdrawal severity but also in the management of chronic ethanol withdrawal induced cognitive impairment.