Ethanol Treatment Research Articles

Hepatoprotective effect of Trigonella foenum graecum against ethanol-induced cell death in human liver cells (HepG2 and Huh7).

The plant Trigonella foenum graecum, also known as fenugreek, has been shown to have anticancer, antidiabetic, anti-inflammatory, and antioxidant properties. In this study, the hepatoprotective effect of fenugreek seed extract (FSE) against ethanol-induced cell death was investigated in human liver cells (HepG2 and Huh7). The cytotoxic effect of FSE and ethanol on cells was evaluated by exposing the cells at different concentrations. Following that, the cells were pre-incubated with 5-25μg/ml FSE, followed by a cytotoxic concentration (0.5mM) of ethanol. MTT and neutral red uptake assays were performed in treated cells to assess the ability of FSE to protect cells from the cytotoxic effects of ethanol. When compared to controls, ethanol treatment significantly reduced the viability of HepG2 and Huh7 cells and altered the cell morphology, whereas treatment with FSE significantly increased cell viability and reversed ethanol-induced morphological changes. Furthermore, pretreatment with FSE dose-dependently reduced lactate dehydrogenate (LDH) leakage, lipid peroxidation (LPO) level, and catalase activities while increasing glutathione (GSH) level induced by ethanol. Pretreatment with FSE also reduced the generation of reactive oxygen species (ROS), caspase enzyme activities, and protein expression of caspase-3 and -9. In HepG2 cells, ethanol-induced apoptosis was observed, whereas FSE treatment reduced apoptosis by downregulating the expression of pro-apoptotic marker genes and upregulating the antiapoptotic gene. In conclusion, this study reports on the mechanistic details of the hepatoprotective potential of FSE. The results also suggest that fenugreek seeds may be useful in preventing liver diseases caused by toxicants such as ethanol.

Effects of Different Treatments on Physicochemical Characteristics of ‘Kyoho’ Grapes during Storage at Low Temperature

Low temperature storage is widely used to maintain the postharvest quality of table grape. However, grape clusters easily undergo deterioration without treatment during the storage time. The main goal of this study was to evaluate the effect of postharvest 1-methylcyclopropene (1-MCP), calcium chloride (1%) and ethanol (16%), and the combination of 1-MCP with calcium chloride and ethanol treatments on maintenance of quality of table grapes ‘Kyoho’ (Vitis vinifera × Vitis labrusca) under 5 °C and 0 °C storage. Changes in decay incidence, weight loss, rachis browning and quality indexes of grape clusters were investigated. The results were as follows: all treatments significantly reduced the decay incidence, weight loss, rachis browning at both low temperatures storage; 1-MCP had positive effect for reducing the decay incidence in early stage, but no effect in late stage; there are no significant variations of taste and color quality indexes under two low temperatures storage, regardless of the treatments. Overall findings suggested that the combination of 1-MCP with calcium chloride and ethanol treatment is suitable for short-term 0 °C storage, while for long-term 0 °C storage, calcium chloride (1%) and ethanol (16%) treatment should be selected.

Ethanol Treatment Enhances Physiological and Biochemical Responses to Mitigate Saline Toxicity in Soybean.

Soil salinity, a major environmental concern, significantly reduces plant growth and production all around the world. Finding solutions to reduce the salinity impacts on plants is critical for global food security. In recent years, the priming of plants with organic chemicals has shown to be a viable approach for the alleviation of salinity effects in plants. The current study examined the effects of exogenous ethanol in triggering salinity acclimatization responses in soybean by investigating growth responses, and numerous physiological and biochemical features. Foliar ethanol application to saline water-treated soybean plants resulted in an enhancement of biomass, leaf area, photosynthetic pigment contents, net photosynthetic rate, shoot relative water content, water use efficiency, and K+ and Mg2+ contents, leading to improved growth performance under salinity. Salt stress significantly enhanced the contents of reactive oxygen species (ROS), malondialdehyde, and electrolyte leakage in the leaves, suggesting salt-induced oxidative stress and membrane damage in soybean plants. In contrast, ethanol treatment of salt-treated soybean plants boosted ROS-detoxification mechanisms by enhancing the activities of antioxidant enzymes, including peroxidase, ascorbate peroxidase, catalase, and glutathione S-transferase. Ethanol application also augmented the levels of proline and total free amino acids in salt-exposed plants, implying a role of ethanol in maintaining osmotic adjustment in response to salt stress. Notably, exogenous ethanol decreased Na+ uptake while increasing K+ and Mg2+ uptake and their partitioning to leaves and roots in salt-stressed plants. Overall, our findings reveal the protective roles of ethanol against salinity in soybean and suggest that the use of this cost-effective and easily accessible ethanol in salinity mitigation could be an effective approach to increase soybean production in salt-affected areas.

Anisotropic silk film inspired by reed leaves for biomedical application

The anisotropic topography of film surfaces plays an important role in regulating cell attachment, morphology, and differentiation. Herein, anisotropic silk film with two different ridge/groove microstructures was created by using reed leaves as templates. The ridge width of silk films derived from the positive and negative of reed leaf were about 279.8 μm and 242.2 μm, respectively. The silk film features a stable β-sheet structure after ethanol treatment. The U87 cells grown on the anisotropic and flat silk film show orientation angles of 13.6° and 46.1°, indicating a directional growth of cells along with the ridge.

A Modified Sequential Deposition Route for High-Performance Carbon-Based Perovskite Solar Cells under Atmosphere Condition.

Carbon-based hole transport material (HTM)-free perovskite solar cells have exhibited a promising commercialization prospect, attributed to their outstanding stability and low manufacturing cost. However, the serious charge recombination at the interface of the carbon counter electrode and titanium dioxide (TiO2) suppresses the improvement in the carbon-based perovskite solar cells’ performance. Here, we propose a modified sequential deposition process in air, which introduces a mixed solvent to improve the morphology of lead iodide (PbI2) film. Combined with ethanol treatment, the preferred crystallization orientation of the PbI2 film is generated. This new deposition strategy can prepare a thick and compact methylammonium lead halide (MAPbI3) film under high-humidity conditions, which acts as a natural active layer that separates the carbon counter electrode and TiO2. Meanwhile, the modified sequential deposition method provides a simple way to facilitate the conversion of the ultrathick PbI2 capping layer to MAPbI3, as the light absorption layer. By adjusting the thickness of the MAPbI3 capping layer, we achieved a power conversation efficiency (PCE) of 12.5% for the carbon-based perovskite solar cells.

Conditioned social preference and reward value of activating oxytocin-receptor-expressing ventral tegmental area neurons following repeated daily binge ethanol intake.

Individuals with alcohol use disorder (AUD) exhibit a disruption of social behavior and dysregulation of oxytocin signaling in the brain, possibly reflecting decreased activation of oxytocin receptors (OxTRs) in reward pathways in response to social stimuli. We hypothesize that daily binge ethanol intake causes a deficit in social reward and oxytocin signaling in the ventral tegmental area (VTA). After 9weeks of daily binge ethanol intake (blood ethanol concentration >80mg%), OxTR-cre mice underwent conditioned place preference for social reward. Separate groups of mice were tested for the effects of binge ethanol on voluntary social interactions, food reward, locomotion, and anxiety-like behaviors. A subset of mice underwent transfection of OxTR-expressing VTA neurons (VTAOxtr ) with a light-sensitive opsin, followed by operant training to respond to light delivered to VTA. Ethanol-naïve male mice increased the time spent on the side previously paired with novel mice while ethanol-treated mice did not. Binge ethanol did not affect conditioned place preference for food reward in males, but this response was weakened in ethanol-treated females. Ethanol treatment also caused a sex-specific impairment of voluntary social interactions with novel mice. There were minimal differences between groups in measures of anxiety and locomotion. Ethanol-naïve mice had significantly greater operant responding for activation of VTAOxtr than sham-transfected mice but ethanol-treated mice did not. There was no difference in the number of VTAOxtr after binge ethanol. Daily binge ethanol causes social reward deficits that cannot be explained by nonspecific effects on other behaviors, at least in males. Only ethanol-naïve mice exhibited positive reinforcement caused by activation of VTAOxtr while daily binge ethanol did not alter the number of VTAOxtr in either males or females. Thus, subtle dysregulation of VTAOxtr function may be related to the social reward deficits caused by daily binge ethanol.

Acute and sub-chronic toxicity study of Rang Chuet (Thunbergia laurifolia Lindl.) extracts and its antioxidant activities

Rang Chuet (Thunbergia laurifolia Lindl.) is a Thai medicinal plant with pharmacological properties: it can be used as an antidote, for decreasing body temperature and it is addictive. This study investigated total phenolic contents and antioxidant activities of Rang Chuet extracts, and its acute and sub-chronic toxicities of Rang Chuet extracts. To investigate the acute toxicity of Rang Chuet, male and female Wistar rats were orally gavaged with a single dose of 2000 and 15,000 mg/kg body weight of Rang Chuet leaf extracts prepared by three different extraction solvents, namely water, ethanol, and acetone. The Rang Chuet water extract showed the highest total phenolic contents at 2643 ± 195.05 mg GAE/100 g while the Rang Chuet acetone extract showed the lowest IC50 at 52.91 mg/ml by DPPH assay. The sub-chronic toxicity study was performed using Wistar rats of both sexes which were gavaged with ethanol and water Rang Chuet extracts for 90 days. Rats were gavaged with the equivalent dose of Rang Chuet in a typical consumer drink (to be taken 3 times a day, at dose 1460 mg/kg/day for water extract and 1025 mg/kg/day for ethanol extract), 3000 and 5000 mg/kg. The satellite group was given the same dose of both extracts for 90 days and observed thereafter for 14 days in order to study the reversibility of the adverse effects. The results revealed that none of the Rang Chuet extracts altered the general behavior or mortality or changes in the gross morphology and any histology of the rats’ visceral organs. For sub-chronic toxicity, the result showed that the treatment of ethanol and water Rang Chuet extracts had no significant effect on average body weight, relative organ weights, histopathology of organs, clinical biochemistry, hematological parameters or liver enzymes. This analysis of by-products of a lipid peroxidation study suggested a trend of decreasing malondialdehyde levels in most of the Rang Chuet treated groups. In conclusion, the safety value of Rang Chuet water extract and Rang Chuet ethanol extract in rats is 50 mg/kg body weight which indicate safe dose of Rang Chuet dried powder is 10.27 g/60 kg body weight per day.

Global transcriptomic analysis of ethanol tolerance response in Salmonella Enteritidis

Adaptation to sublethal amounts of ethanol enables Salmonella Enteritidis to survive under normally lethal ethanol conditions, which is referred to as the ethanol tolerance response (ETR). To uncover mechanisms underlying this adaptative response, RNA-seq and RT-qPCR techniques were employed to reveal global gene expression patterns in S. Enteritidis after sublethal ethanol treatment. It was observed that 811 genes were significantly differentially expressed in ethanol-treated cells compared with control cells, among which 328 were up-regulated and 483 were down-regulated. Functional analysis revealed that these genes were enriched in different pathways, including signal transduction, membrane transport, metabolism, transcription, translation, and cell motility. Specifically, a couple of genes encoding histidine kinases and response regulators in two-component systems were up-regulated to activate sensing and signaling pathways. Membrane function was also influenced by ethanol treatment since ABC transporter genes for transport of glutamate, phosphate, 2-aminoethylphosphonate, and osmoprotectant were up-regulated, while those for transport of iron complex, manganese, and ribose were down-regulated. Accompanied with this, diverse gene expression alterations related to the metabolism of amino acids, carbohydrates, vitamins, and nucleotides were observed, which suggested nutritional requirements for S. Enteritidis to mount the ETR. Furthermore, genes associated with ribosomal units, bacterial chemotaxis, and flagellar assembly were generally repressed as a possible energy conservation strategy. Taken together, this transcriptomic study indicates that S. Enteritidis employs multiple genes and adaptation pathways to develop the ETR.

Structural changes and exposed amino acids of ethanol-modified whey proteins isolates promote its antioxidant potential

Whey protein isolates (WPI) were treated with different ethanol level (20, 40, 60, and 80%, v/v) to promote structural unfolding and subsequent aggregation. In general, protein aggregation gradually increased with increasing ethanol level in a dose-dependent manner, which was implied by notably increased turbidity and gradually decreased solubility. The formation of aggregates, which were confirmed by the results of circular dichroism spectrum and total sulfhydryl content, were promoted mainly through disulfide bonds and intra-molecular hydrogen bonds. Moreover, ethanol treated WPI (E-WPI) had significantly enhanced antioxidant activities over native WPI, which was mainly attribute to the higher contents of specific amino acids (such as hydrophobic amino acids, aromatic amino acids, and sulfur-containing amino acids), and E-WPI prepared with moderate ethanol concentration (40% in our present study) exhibited the highest antioxidant activities. These results reveal that antioxidant activities of WPI can be increased by ethanol treatment and are possibly achieved through molecular unfolding of native WPI.

Removal of anti-nutritional factors of rapeseed protein isolate (RPI) and toxicity assessment of RPI.

We prepared a detoxified rapeseed protein isolate (RPI) by phytase/ethanol treatment based on alkaline extraction and acidic precipitation. Contents of protein, fat, ash, moisture, crude fiber, glucosinolates, phytic acid, and phenolics and color were determined. To evaluate the safety of detoxified RPI, five groups of C57 mice (detoxified RPI [10 and 20 g kg-1]; commercial soybean protein isolate (SPI) [10 g kg-1]; non-detoxified RPI [10 g kg-1]; control) were used in the acute-toxicity test. Bodyweight and pathology parameters were recorded at different time points, followed by macroscopic examination, organ-weight measurement and microstructure examination. After pretreatment of rapeseed meals with phytase (enzyme : substrate ratio, 1 : 5 mg g-1) for 1.5 h and two-time ethanol extraction for precipitated protein, the chemical characteristics in RPI were protein (88.26%), fat (0.57%), ash (2.72%), moisture (1.90%), crude fiber (0.77%), glucosinolates (0 μmol g-1), phytic acid (0.17%), phenolics (0.36%) and whiteness (73.38). Treatment resulted in significant removal of anti-nutritional factors (ANFs) and increased whiteness in detoxified RPI compared with non-detoxified RPI, and lower than in cruciferin-rich canola protein isolate (Puratein®). Experimental-related effects on bodyweight, clinical observations, or clinicopathology, in mice treated with detoxified RPI were not observed except for a decreased thyroid gland/parathyroid gland index in mice treated with non-detoxified RPI. Furthermore, the no-observed-effect level (NOEL) was 10 g kg-1 of detoxified RPI, whereas the no-observed-adverse-effect-level (NOAEL) was the highest fed level of 20 g kg-1 of detoxified RPI. Overall, detoxified RPI prepared by the combined treatment of phytase and ethanol was considered safe under the conditions tested, in which the contents of the main ANFs were reduced significantly.

Intraoperative 96% ethanol as an adjuvant in Pterygium (Pt) surgery

AbstractPurposeThe aim of the study is to present how we use ethanol on the cornea next to the pterygium head to remove the corneal epithelium and create a dissection plane to remove the fibrovascular tissue in a more controlled way leaving a more regular corneal surface.MethodsTopical tetracaine + oxybuprocaine as topical anaesthetic, povidone iodine 5% is instilled and mepivacaine 2% is injected under the body of the pterygium. A cellulose sponge soaked in 96% ethanol is placed over the corneal surface 1‐2 mm around the head of the fibrovascular tissue for 10 seconds, then washed. Blunt lifting of the epithelium towards the head of the pterygium allows identification of the surgical plane for dissection of the underlying corneal adhesions. Excision of the corneal encroaching tissue, tennon's capsule over the bare sclera and under the conjunctival wound was followed by conjunctival autograft sealed with fibrin (tisseel).ResultsA retrospective review of all pterygium procedures performed at our institution from January 2017 to January 2020 was performed to find out if there were any intra‐ or post‐surgical complications related to the use of 96% ethanol. All patients had completed epithelialisation within 1 week and none had limbal deficiency. The recurrence rate was very low.ConclusionsIntrasurgical application of 96% ethanol on the cornea next to the pterygium head for 10 seconds just before surgical removal of the pterygium is an inexpensive, safe and effective adjuvant to the procedure. It may even be of more interest in recurrent pterygium where firm adhesions need to be addressed and creating a new smooth surface may prevent recurrences.BibliographyKaufman SC, Jacobs DS, Lee WB, Deng SX, Rosenblatt MI, Shtein RM. Options and adjuvants in surgery for pterygium: a report by the American Academy of Ophthalmology. Ophthalmology. 2013 Jan;120(1):201‐8. https://doi.org/10.1016/j.ophtha.2012.06.066. Epub 2012 Oct 11. PMID: 23062647.Chen KH, Hsu WM. Intraoperative ethanol treatment as an adjuvant therapy of pterygium excision. Int J Biomed Sci. 2006 Dec;2(4):414‐21. PMID: 23675010; PMCID: PMC3614651.Tsumi E, Levy J, Pitchkhadze A, Baidousi A, Lifshitz T. New approach for pterygium removal using 20 % ethanol. Int Ophthalmol. 2012 Oct;32(5):443‐8. https://doi.org/10.1007/s10792‐012‐9597‐2. Epub 2012 Jun 21. PMID: 22717949.

Mechanisms of Ethanol Treatment on Maintaining Quality and Controlling Browning in Fresh-Cut Lotus Roots

Mechanisms of Ethanol Treatment on Maintaining Quality and Controlling Browning in Fresh-Cut Lotus Roots

Hepatocyte-specific deletion of cellular repressor of E1A-stimulated genes 1 exacerbates alcohol-induced liver injury by activating stress kinases.

Alcohol-associated liver disease (ALD) encompasses a wide range of pathologies from simple steatosis to cirrhosis and hepatocellular carcinoma and is a global health problem. Currently, there are no effective pharmacological treatments for ALD. We have previously demonstrated that aging exacerbates the pathogenesis of ALD, but the underlying mechanisms are still poorly understood. Cellular repressor of E1A-stimulated genes 1 protein (CREG1) is a recently identified small glycoprotein that has been implicated in aging process by promoting cellular senescence and activating stress kinases. Thus, the current study aimed to explore the role of aging associated CREG1 in ALD pathogenesis and CREG1 as a potential therapeutic target. Hepatic and serum CREG1 protein levels were elevated in ALD patients. Elevation of hepatic CREG1 protein and mRNA was also observed in a mouse model of Gao-binge alcohol feeding. Genetic deletion of the Creg1 gene in hepatocytes (Creg1∆hep) markedly exacerbated ethanol-induced liver injury, apoptosis, steatosis and inflammation. Compared to wild-type mice, Creg1∆hep mice had increased phosphorylation of hepatic stress kinases such as apoptosis signal-regulating kinase 1 (ASK1), c-Jun N-terminal kinase (JNK) and p38 but not TGF-β-activated kinase 1 (TAK1) or extracellular signal-regulated kinase (ERK) after alcohol feeding. In vitro, ethanol treatment elevated the phosphorylation of ASK1, JNK, and p38 in mouse hepatocyte AML-12 cells. This elevation was further enhanced by CREG1 knockdown but alleviated by CREG1 overexpression. Last, treatment with an ASK1 inhibitor abolished ethanol-induced liver injury and upregulated hepatic lipogenesis, proinflammatory genes and stress kinases in Creg1∆hep mice. Taken together, our data suggest that CREG1 protects against alcoholic liver injury and inflammation by inhibiting the ASK1-JNK/p38 stress kinase pathway and that CREG1 is a potential therapeutic target for ALD.

Oil Recovery from Dry Grind Ethanol Plant Coproducts Using Ethanol

Corn ethanol bio-refineries are seeking economic processing strategies for recovering oil from their coproducts. The addition of ethanol can be an efficient method to recover the oil from the coproducts as the industry has available ethanol. This study considered the effects of ethanol on oil recovery from distillers’ dried grains with solubles (DDGS) and oil partitioning from whole stillage (WS) on a laboratory scale. Ethanol was added with original and heavier fraction DDGS in different temperatures (room temperature ~20 °C, 30 °C, 40 °C, and 50 °C) and solids loadings (20%, 30%, and 40%), and their effects on oil recovery were evaluated. The whole stillage was incubated with ethanol at room temperature (~20 °C) and 50 °C separately to analyze WS’s oil distribution in the liquid and solid phases. The amount of recovered oil from the original and heavier fractions of DDGS varies from 25–45% and 45–70%, respectively, with an increment of temperature. Increasing solids loadings up to 30% had no effect on oil recovery from either DDGS sample. Ethanol treatment in WS resulted in 8–10% higher wet yield of liquid fraction and 17–20% of oil increase in liquid fraction than the control treatment. It is also notable that temperature positively impacted oil partitioning from WS. The results showed that ethanol could improve oil recovery from DDGS and oil partition in WS by varying different process conditions. This outcome is beneficial to ethanol plants to increase corn oil yield using their existing setup and in-situ product.

Effect of ethanol treatment on morphological and technical properties of corn starch

Solvent exchange is considered an effective method that changes the physicochemical properties of starch, especially the absorption. Ethanol concentration is one of the important influencing factors to form porous pores from the surface to inside starch granules. In this study, the effect of ethanol concentration on technical properties of starch treated by solvents were investigated.
 The ratio of ethanol/water was prepared at 1/9, 3/7, 5/5, 7/3 and 10/0 (w/w), respectively. Corn starch was treated by solvent at 8% concentration. Morphology, oil and water holding capacity, solubility, swelling power, viscosity, and transmittance were studied to elucidate the effect of the ethanol concentration on the morphological and technical properties of corn starch treated by solvent. As a result, starch treated by solvent with different concentrations tended to form wrinkles and pore from the surface to the inside of the starch granules during ethanol immersion. Therefore, the oil and water holding capacity, solubility, swelling and viscosity of the solvent-treated starch samples increase as the concentration of ethanol increases. Otherwise, the transmittance of starch glues tends to decrease when the ethanol concentration increases.

Protein Enrichment of Wheat Bread with Microalgae: Microchloropsis gaditana, Tetraselmis chui and Chlorella vulgaris.

Cell wall disrupted and dried Microchloropsis gaditana (Mg), Tetraselmis chui (Tc) and Chlorella vulgaris (Cv) microalgae biomasses, with or without ethanol pre-treatment, were added to wheat bread at a wheat flour substitution level of 12%, to enrich bread protein by 30%. Baking performance, protein quality and basic sensory properties were assessed. Compared to wheat, Mg, Tc and Cv contain higher amounts of essential amino acids and their incorporation markedly improved protein quality in the bread (DIAAS 57–66 vs. 46%). The incorporation of microalgae reduced dough strength and bread volume and increased crumb firmness. This was most pronounced for Cv and Tc but could be improved by ethanol treatment. Mg gave adequate dough strength, bread volume and crumb structure without ethanol treatment. To obtain bread of acceptable smell, appearance, and colour, ethanol treatment was necessary also for Mg as it markedly reduced the unpleasant smell and intense colour of all algae breads. Ethanol treatment reduced the relative content of lysine, but no other essential amino acids. However, it also had a negative impact on in vitro protein digestibility. Our results show that Mg had the largest potential for protein fortification of bread, but further work is needed to optimize pre-processing and assess consumer acceptance.

Hydro- and aerogels from ethanolic potato and whey protein solutions: Influence of temperature and ethanol concentration on viscoelastic properties, protein interactions, and microstructure

Denaturation, aggregation, and gelation of protein solutions can be induced through increased temperatures and the presence of organic solvents. Depending on the molecular features of the involved proteins, the denaturation conditions lead to very different gel properties. However, it is poorly understood how organic solvents in combination with heat treatments can be used to modify the textural properties of protein gels. In this study, the combined effects of heat and ethanol (EtOH) treatment on hydrogel formation by whey (WPI) and potato protein isolates (PPI) were investigated. The different protein hydrogels were subjected to a complete solvent exchange with EtOH and dried with supercritical CO2 to obtain aerogels with a high inner surface area. Increasing EtOH concentration during thermally induced hydrogel formation reduced the temperature of unfolding up to a point where denaturation occurred at room temperature. WPI and PPI formed very different gels in the presence of EtOH. WPI gels with an EtOH content below 10% (w/w) were elastic and mainly linked through disulfide bonds. Higher EtOH content led to weak gels, mainly linked through electrostatic and hydrogen bonds. For PPI gels, increasing EtOH content did not influence the protein interactions within the gels, and textural properties were very similar. The textural properties of the resulting aerogels were dependent on the type of protein interactions created in the hydrogels. This work provides insights into the way food proteins form gel networks and how these interactions can be manipulated to produce gels with tailor-made properties.

Ethanol exposure of human pancreatic normal ductal epithelial cells induces EMT phenotype and enhances pancreatic cancer development in KC (Pdx1‐Cre and LSL‐KrasG12D) mice

Alcohol is a risk factor for pancreatic cancer. However, the molecular mechanism by which chronic alcohol consumption influences pancreatic cancer development is not well understood. We have recently demonstrated that chronic ethanol exposure of pancreatic normal ductal epithelial cells (HPNE) induces cellular transformation by generating cancer stem cells (CSCs). Here, we examined whether chronic ethanol treatment induces epithelial–mesenchymal transition in HPNE cells and promotes pancreatic cancer development in KC (Pdx1‐Cre, and LSL‐KrasG12D) mice. Our data demonstrate that chronic ethanol exposure of HPNE cells induces SATB2 gene and those cells became highly motile. Ethanol treatment of HPNE cells results in downregulation of E‐Cadherin and upregulation of N‐Cadherin, Snail, Slug, Zeb1, Nanog and BMI‐1. Suppression of SATB2 expression in ethanol‐transformed HPNE cells inhibits EMT phenotypes. KC mice fed with an ethanol‐containing diet show enhanced pancreatic cancer growth and development than those fed with a control diet. Pancreas isolated from KC mice fed with an ethanol‐containing diet show higher expression of stem cell markers (CD133, CD44, CD24), pluripotency‐maintaining factors (cMyc, KLF4, SOX‐2, and Oct‐4), N‐Cadherin, EMT‐transcription factors (Snail, Slug, and Zeb1), and lower expression of E‐cadherin than those isolated from mice fed with a control diet. Furthermore, pancreas isolated from KC mice fed with an ethanol‐containing diet show higher expression of inflammatory cytokines (TNF‐α, IL‐6, and IL‐8) and PTGS‐2 (COX‐2) gene than those isolated from mice fed with a control diet. These data suggest that chronic alcohol consumption may contribute to pancreatic cancer development by generating inflammatory signals and CSCs.

Neonatal ethanol causes profound reduction of cholinergic cell number in the basal forebrain of adult animals.

In animal models that mimic human third-trimester fetal development, ethanol causes substantial cellular apoptosis in the brain, but for most brain structures, the extent of permanent neuron loss that persists into adulthood is unknown. We injected ethanol into C57BL/6J mouse pups at postnatal day 7 (P7) to model human late-gestation ethanol toxicity, and then used stereological methods to investigate adult cell numbers in several subcortical neurotransmitter systems that project extensively in the forebrain to regulate arousal states. Ethanol treatment caused especially large reductions (34-42%) in the cholinergic cells of the basal forebrain, including cholinergic cells in the medial septal/vertical diagonal band nuclei (Ch1/Ch2) and in the horizontal diagonal band/substantia innominata/nucleus basalis nuclei (Ch3/Ch4). Cell loss was also present in non-cholinergic basal forebrain cells, as demonstrated by 34% reduction of parvalbumin-immunolabeled GABA cells and 25% reduction of total Nissl-stained neurons in the Ch1/Ch2 region. In contrast, cholinergic cells in the striatum were reduced only 12% by ethanol, and those of the brainstem pedunculopontine/lateral dorsal tegmental nuclei (Ch5/Ch6) were not significantly reduced. Similarly, ethanol did not significantly reduce dopamine cells of the ventral tegmental area/substantia nigra or serotonin cells in the dorsal raphe nucleus. Orexin (hypocretin) cells in the hypothalamus showed a modest reduction (14%). Our findings indicate that the basal forebrain is especially vulnerable to alcohol exposure in the late gestational period. Reduction of cholinergic and GABAergic projection neurons from the basal forebrain that regulate forebrain arousal may contribute to the behavioral and cognitive deficits associated with neonatal ethanol exposure.

Facile Fabrication of Silk Fibroin/Konjac Glucomannan Composite Membranes

Silk fibroin (SF)-based biocomposites have attracted considerable attention for several decades, due to excellent biocompatibility and mechanical toughness. However, insufficient hydrophilicity and mechanical brittleness limit its practical application, especially in tissue engineering. In this study, by blending konjac glucomannan (KGM) with SF, along with the water-insoluble treatment of ethanol and ammonia, SF/KGM composite membranes were generated with good flexibility and water adsorption capacity. Scanning electron microscopy (SEM) showed that KGM/SF membrane surfaces were uniform and dense. Fourier transform infrared spectroscopy (FTIR) showed that the silk II structure was dominant. Contact angle and mechanical testing showed that the use of KGM with SF in membranes helped to form more flexible and hydrophilic membranes for potential use in biomaterials and devices.