Maltose Dextrin Research Articles

In Vivo Administration of Phosphatidic Acid, a Direct Alcohol Target Rescues Fetal Growth Restriction and Maternal Uterine Artery Dysfunction in Rat FASD Model.

Fetal growth restriction is a hallmark of Fetal Alcohol Syndrome (FAS) and is accompanied by maternal uterine circulatory maladaptation. FAS is the most severe form of Fetal Alcohol Spectrum Disorder (FASD), a term for the range of conditions that can develop in a fetus when their pregnant mother consumes alcohol. Alcohol exerts specific direct effects on lipids that control fundamental developmental processes. We previously demonstrated that direct in vitro application of phosphatidic acid (PA, the simplest phospholipid and a direct target of alcohol exposure) to excised uterine arteries from alcohol-exposed rats improved vascular function, but it is unknown if PA can rescue end organ phenotypes in our FASD animal model. Pregnant Sprague-Dawley rats (n = 40 total dams) were gavaged daily from gestational day (GD) 5 to GD 19 with alcohol or maltose dextrin, with and without PA supplementation, for a total of four unique groups. To translate and assess the beneficial effects of PA, we hypothesized that in vivo administration of PA concomitant with chronic binge alcohol would reverse uterine artery dysfunction and fetal growth deficits in our FASD model. Mean fetal weights and placental efficiency were significantly lower in the binge alcohol group compared with those in the control (p < 0.05). However, these differences between the alcohol and the control groups were completely abolished by auxiliary in vivo PA administration with alcohol, indicating a reversal of the classic FAS growth restriction phenotype. Acetylcholine (ACh)-induced uterine artery relaxation was significantly impaired in the uterine arteries of chronic in vivo binge alcohol-administered rats compared to the controls (p < 0.05). Supplementation of PA in vivo throughout pregnancy reversed the alcohol-induced vasodilatory deficit; no differences were detected following in vivo PA administration between the pair-fed control and PA alcohol groups. Maximal ACh-induced vasodilation was significantly lower in the alcohol group compared to all the other treatments, including control, control PA, and alcohol PA groups (p < 0.05). When analyzing excitatory vasodilatory p1177-eNOS, alcohol-induced downregulation of p1177-eNOS was completely reversed following in vivo PA supplementation. In summary, these novel data utilize a specific alcohol target pathway (PA) to demonstrate a lipid-based preventive strategy and provide critical insights important for the development of translatable interventions.

Sex Differences in Alcohol-related Cardiomyopathy

Alcohol-related Cardiomyopathy (ACM) manifests in humans with a significant history of alcohol use and is characterized by ventricular dilation and cardiac function impairment. Although the most common age group for ACM is males between the ages of 30-55, women require a lesser lifetime alcohol exposure to develop ACM than men. The objective of this study is to assess sexual differences in a mouse model of chronic plus binge alcohol consumption. We hypothesize that female mice exposed to chronic plus binge alcohol will not develop as severe cardiac dysfunction as their male counterparts. In this study, male (n=19) and female (n=14) C57BL/6J mice were provided chronic plus binge alcohol feeding. After 5 days of acclimation to the liquid diet, mice are fed ad libitum 5% ethanol (EtOH) liquid or isocaloric control liquid diet for 30 days. At 10 and 30 days, mice received an oral binge dose of EtOH (5 g/kg body weight), or isocaloric maltose dextrin solution (9 g/kg body weight). Noninvasive cardiac structural and functional data were obtained via echocardiography at baseline and following binges. After 30 days, cardiac hemodynamics was measured by open chest left ventricle catheterization. Two-way ANOVA with Tukey’s post hoc analysis was performed for each cardiac parameter, and significance was defined as p&lt;0.05. Male EtOH mice exhibited a significant decrease in stroke work (1119 ± 297 mmHg/mL when compared to male controls, 1622 ± 365 mmHg/mL; p= 0.03). Female EtOH mice had a nonsignificant decrease in stroke work (1242 ± 374 mmHg/mL when compared to female controls, 1423 ± 469 mmHg/mL; p= 0.80). Maximum left ventricular pressure was decreased 20% in male EtOH mice (p=0.003) but increased 10% in female EtOH mice (p=0.46). The maximum rate of pressure change within the ventricle was 24% lower in the male EtOH mice (p=0.02) and 9% higher in female EtOH mice (p=0.87). The minimum rate of pressure change within the ventricle was 24% higher in male EtOH mice (p=0.02) and 17% lower in female ETOH mice (p=0.44). After 30 days of EtOH exposure, male mice showed significant systolic and diastolic dysfunction. Female mice on the other hand, did not show substantial cardiac dysfunction after chronic plus binge ethanol exposure. Further experiments involving qPCR of proinflammatory and profibrotic markers will elucidate the molecular mechanism of sex differences in ACM development. R21AA029747 (JG &amp; CB) &amp; T32AA007577. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

MiR-141/200c contributes to ethanol-mediated hepatic glycogen metabolism

ObjectiveHepatic glucose metabolism is profoundly perturbed by excessive alcohol intake. miR-141/200c expression is significantly induced by chronic ethanol feeding. This study aimed at identifying the role of miR-141/200c in glucose homeostasis during chronic ethanol exposure. MethodsWT and miR-141/200c KO mice were fed a control or an ethanol diet for 30 days, followed by a single binge of maltose dextrin or ethanol, respectively. Untargeted metabolomics analysis of hepatic primary metabolites was performed along with analyses for liver histology, gene expression, intracellular signaling pathways, and physiological relevance. Primary hepatocytes were used for mechanistic studies. ResultsmiR-141/200c deficiency rewires hepatic glucose metabolism during chronic ethanol feeding, increasing the abundance of glucose intermediates including G6P, an allosteric activator for GS. miR-141/200c deficiency replenished glycogen depletion during chronic ethanol feeding accompanied by reduced GS phosphorylation in parallel with increased expression of PP1 glycogen targeting subunits. Moreover, miR-141/200c deficiency prevented ethanol-mediated increases in AMPK and CaMKK2 activity. Ethanol treatment reduced glycogen content in WT-hepatocytes, which was reversed by dorsomorphin, a selective AMPK inhibitor, while KO-hepatocytes displayed higher glycogen content than WT-hepatocytes in response to ethanol treatment. Furthermore, treatment of hepatocytes with A23187, a calcium ionophore activating CaMKK2, lowered glycogen content in WT-hepatocytes. Notably, the suppressive effect of A23187 on glycogen deposition was reversed by dorsomorphin, demonstrating that the glycogen depletion by A23187 is mediated by AMPK. KO-hepatocytes exhibited higher glycogen content than WT-hepatocytes in response to A23187. Finally, miR-141/200c deficiency led to improved glucose tolerance and insulin sensitivity during chronic ethanol feeding. ConclusionsmiR-141/200c deficiency replenishes ethanol-mediated hepatic glycogen depletion through the regulation of GS activity and calcium signaling coupled with the AMPK pathway, improving glucose homeostasis and insulin sensitivity. These results underscore miR-141/200c as a potential therapeutic target for the management of alcohol intoxication.

Liposomal Glutathione Augments Immune Defenses against Respiratory Syncytial Virus in Neonatal Mice Exposed in Utero to Ethanol.

We previously reported that maternal alcohol use increased the risk of sepsis in premature and term newborns. In the neonatal mouse, fetal ethanol (ETOH) exposure depleted the antioxidant glutathione (GSH), which promoted alveolar macrophage (AM) immunosuppression and respiratory syncytial virus (RSV) infections. In this study, we explored if oral liposomal GSH (LGSH) would attenuate oxidant stress and RSV infections in the ETOH-exposed mouse pups. C57BL/6 female mice were pair-fed a liquid diet with 25% of calories from ethanol or maltose-dextrin. Postnatal day 10 pups were randomized to intranasal saline, LGSH, and RSV. After 48 h, we assessed oxidant stress, AM immunosuppression, pulmonary RSV burden, and acute lung injury. Fetal ETOH exposure increased oxidant stress threefold, lung RSV burden twofold and acute lung injury threefold. AMs were immunosuppressed with decreased RSV clearance. However, LGSH treatments of the ETOH group normalized oxidant stress, AM immune phenotype, the RSV burden, and acute lung injury. These studies suggest that the oxidant stress caused by fetal ETOH exposure impaired AM clearance of infectious agents, thereby increasing the viral infection and acute lung injury. LGSH treatments reversed the oxidative stress and restored AM immune functions, which decreased the RSV infection and subsequent acute lung injury.

Effects of moderate ethanol consumption as a function of n-6:n-3 dietary ratio on lipid profile, inflammation, and liver function in mice

ObjectiveIt is critical to understand how moderate ethanol exposure interacts with dietary components such as essential fatty acids to influence inflammatory processes underlying CVD pathogenesis. The purpose of this study was to examine the effects of moderate ethanol consumption and dietary n-6:n-3 fatty acid composition on markers associated with CVD in mice. MethodsTwenty-three C57BL/6J mice consumed an 18% ethanol solution or 26.9% maltose dextrin solution (isocaloric control) for 12 weeks. Within each group, the mice were fed either a high n-6 (n-6:n-3 = 50:1) diet or a balanced n-3 (n-6:n-3 = 1:1) diet ad libitum. Following the exposure period, serum samples were analyzed to assess lipid profile, inflammatory markers, antioxidant capacity, DNA damage, and liver function enzyme activity. ResultsThe control group gained more weight than the ethanol group (P = 0.020). In ethanol-exposed mice, HDL was significantly increased (P = 0.009). C-reactive protein (CRP; P < 0.001), high mobility group box 1 protein (HMGB1; P = 0.011), 8-oxo-deoxyguanosine (8-oxo-dG; P = 0.019), ALT (P = 0.002) and AP (P = 0.021) were lower in the ethanol group. There was a significant main effect of the n-3 diet on total antioxidant capacity (TAC; P < 0.001) and 8-oxo-dG (P = 0.047). ConclusionThese findings indicate that moderate ethanol consumption and a balanced n-6:n-3 diet improve several inflammatory and lipid markers associated with CVD. Observed differences in weight gain between groups should be considered when interpreting these results.

IN SILICO PROOFS FOR PHLORIDZIN, NARINGENIN, AND CINNAMIC ACID AS ALPHA-AMYLASE ACTIVATORS, WHICH IS IMPORTANT IN INDUSTRIAL MICROBIOLOGY OR BIOCHEMICAL ENGINEERING

Enzymes are commonly defined as biological catalysts, regulating particular biochemical reactions. α-Amylase (EC 3.2.1.1) is one of the industrially important enzymes, which are extensively used in starch hydrolyzing processes, such as brewing, fermentation, detergent production, food processing, etc. This enzyme breaks down α-1,4 glycosidic bonds in amylose or amylopectin. The end products from amylose are maltotriose and maltose. Maltose, glucose, and limit dextrin are formed from amylopectin. There are many studies in the literature regarding the α-amylase inhibitors, which have the potentials of being used in diabetes and obesity. However, there is a very limited number of studies in the literature about the activation of this enzyme, which could be harmful to such diseases. This study aims to support the activation activity of phloridzin, naringenin, and cinnamic acid for α-amylase, which was previously proved experimentally, with some in silico tests.

Morphological alteration in rat hippocampal neuronal dendrites following chronic binge prenatal alcohol exposure

Prenatal alcohol exposure (PAE) may result in Fetal Alcohol Spectrum Disorders (FASD). The hippocampus has been recognized as a vulnerable target to alcohol-induced developmental damage. However, the effect of prenatal exposure to alcohol on dendritic morphological adaptations throughout the hippocampal fields in the developing brain still remains largely unknown in the context of FASD. We hypothesized that chronic binge alcohol exposure during pregnancy alters dendrite arborization throughout the developing rat hippocampus. Pregnant Sprague-Dawley rats were assigned to either a pair-fed control (PF-Cont) or a binge alcohol (Alcohol) treatment group. Alcohol dams were acclimatized via a once-daily orogastric gavage of 4.5 g/kg alcohol from gestational day (GD) 5–10 and progressed to 6 g/kg alcohol from GD 11–21. Pair-fed dams similarly received isocaloric maltose dextrin. After parturition, all dams received an ad libitum diet and nursed their offspring until postnatal day (PND) 10 when the pup brains were collected for morphological analysis. PAE increased dendritic arborization and complexities of CA1, CA2/3, and DG neurons in the PND 10 rat hippocampus. The number of primary dendrites, total dendritic length, and number of dendritic branches were significantly increased following PAE, and Sholl analysis revealed significantly more intersections of the dendritic processes in PND 10 offspring following PAE compared with those in the PF-Cont group. We conclude that chronic binge PAE significantly alters hippocampal dendritic morphology in the developing hippocampus. We conjecture that this morphological alteration in postnatal rat hippocampal dendrites following chronic binge prenatal alcohol exposure may play a critical role in FASD neurobiological phenotypes.

Effects of Moderate Ethanol Consumption with a n‐3 Diet on Inflammation and Total Antioxidant Capacity in Mice

Excessive alcohol intake is known to elevate risk for cardiovascular diseases (CVD); however, several studies have reported that moderate alcohol consumption may not contribute additional risk or may provide a protective effect against CVD. It is critical to understand how moderate ethanol exposure interacts with other essential dietary components such as n-3 fatty acids to influence inflammation underlying CVD pathogenesis. The purpose of this study was to examine the effects of moderate ethanol consumption as a function of dietary n-6:n-3 fatty acid composition on markers associated with inflammation and total antioxidant capacity (TAC) in mice. Twenty-three mice (12 male, 11 female) consumed an 18% ethanol solution or 26.9% maltose dextrin solution (non-ethanol isocaloric control) for 12 weeks. In each group, half of the mice were fed either a high n-6 (n-6:n-3 = 50:1) diet or a balanced n-3 (n-6:n-3 = 1:1) diet ad libitum. There were no differences in initial body weight between the two groups; however, the control group gained significantly more weight than the ethanol group (P = 0.020) associated with higher maltose dextrin fluid intake (P < 0.001). C-reactive protein and high mobility group box 1 protein were significantly lower in the ethanol group compared to the control group (P < 0.001 and P = 0.029, respectively). There was a significant main effect of n-3 diet on total antioxidant capacity (TAC; P < 0.001) and no significant effect of ethanol on TAC. These findings indicate that moderate consumption of alcohol may improve inflammatory markers associated with CVD in mice; however, the results should be interpreted with caution due to the differences in weight gain between groups.

Oyster hydrolysate ameliorates ethanol diet‐induced alcoholic fatty liver by regulating lipid metabolism in rats

SummaryWe evaluated the effect of oyster hydrolysate, which was prepared using transglutaminase, protamex and neutrase (TGPN), on alcoholic fatty liver. Sprague–Dawley rats were fed a diet containing 5% (v/v) ethanol or an isocaloric amount of dextrin–maltose for 6 weeks and orally administered TGPN or silymarin for another 4 weeks. TGPN significantly decreased the liver index (liver weight/body weight), serum and hepatic lipid levels and increased the serum adiponectin level. Based on histological analysis, TGPN reduced the number of lipid droplets and lipid accumulation in liver tissue. TGPN enhanced the expression of AMP‐activated protein kinase and activated its downstream pathway which controls lipid metabolism. Additionally, TGPN decreased the levels of liver function markers (aspartate aminotransferase and alanine aminotransferase) and tumour necrosis factor alpha. Consequently, TGPN ameliorated alcoholic fatty liver by inhibiting fatty acid and cholesterol synthesis, promoting fatty acid oxidation and decreasing liver cell damage.

Aspects and Recent Trends in Microbial α-Amylase: a Review.

α-Amylases are the oldest and versatile starch hydrolysing enzymes which can replace chemical hydrolysis of starch in industries. It cleaves the α-(1,4)-D-glucosidic linkage of starch and other related polysaccharides to yield simple sugars like glucose, maltose and limit dextrin. α-Amylase covers about 30% shares of the total enzyme market. On account of their superior features, α-amylase is the most widely used among all the existing amylases for hydrolysis of polysaccharides. Endo-acting α-amylase of glycoside hydrolase family 13 is an extensively used biocatalyst and has various biotechnological applications like in starch processing, detergent, textile, paper and pharmaceutical industries. Apart from these, it has some novel applications including polymeric material for drug delivery, bioremediating agent, biodemulsifier and biofilm inhibitor. The present review will accomplish the research gap by providing the unexplored aspects of microbial α-amylase. It will allow the readers to know about the works that have already been done and the latest trends in this field. The manuscript has covered the latest immobilization techniques and the site-directed mutagenesis approaches which are readily being performed to confer the desirable property in wild-type α-amylases. Furthermore, it will state the inadequacies and the numerous obstacles coming in the way of its production during upstream and downstream steps and will also suggest some measures to obtain stable and industrial-grade α-amylase.

New strain of Pediococcus pentosaceus alleviates ethanol-induced liver injury by modulating the gut microbiota and short-chain fatty acid metabolism.

BACKGROUNDIntestinal dysbiosis has been shown to be associated with the pathogenesis of alcoholic liver disease (ALD), which includes changes in the microbiota composition and bacterial overgrowth, but an effective microbe-based therapy is lacking. Pediococcus pentosaceus (P. pentosaceus) CGMCC 7049 is a newly isolated strain of probiotic that has been shown to be resistant to ethanol and bile salts. However, further studies are needed to determine whether P. pentosaceus exerts a protective effect on ALD and to elucidate the potential mechanism.AIMTo evaluate the protective effect of the probiotic P. pentosaceus on ethanol-induced liver injury in mice.METHODSA new ethanol-resistant strain of P. pentosaceus CGMCC 7049 was isolated from healthy adults in our laboratory. The chronic plus binge model of experimental ALD was established to evaluate the protective effects. Twenty-eight C57BL/6 mice were randomly divided into three groups: The control group received a pair-fed control diet and oral gavage with sterile phosphate buffered saline, the EtOH group received a ten-day Lieber-DeCarli diet containing 5% ethanol and oral gavage with phosphate buffered saline, and the P. pentosaceus group received a 5% ethanol Lieber-DeCarli diet but was treated with P. pentosaceus. One dose of isocaloric maltose dextrin or ethanol was administered by oral gavage on day 11, and the mice were sacrificed nine hours later. Blood and tissue samples (liver and gut) were harvested to evaluate gut barrier function and liver injury-related parameters. Fresh cecal contents were collected, gas chromatography–mass spectrometry was used to measure short-chain fatty acid (SCFA) concentrations, and the microbiota composition was analyzed using 16S rRNA gene sequencing.RESULTSThe P. pentosaceus treatment improved ethanol-induced liver injury, with lower alanine aminotransferase, aspartate transaminase and triglyceride levels and decreased neutrophil infiltration. These changes were accompanied by decreased levels of endotoxin and inflammatory cytokines, including interleukin-5, tumor necrosis factor-α, granulocyte colony-stimulating factor, keratinocyte-derived protein chemokine, macrophage inflammatory protein-1α and monocyte chemoattractant protein-1. Ethanol feeding resulted in intestinal dysbiosis and gut barrier disruption, increased relative abundance of potentially pathogenic Escherichia and Staphylococcus, and the depletion of SCFA-producing bacteria, such as Prevotella, Faecalibacterium, and Clostridium. In contrast, P. pentosaceus administration increased the microbial diversity, restored the relative abundance of Lactobacillus, Pediococcus, Prevotella, Clostridium and Akkermansia and increased propionic acid and butyric acid production by modifying SCFA-producing bacteria. Furthermore, the levels of the tight junction protein ZO-1, mucin proteins (mucin [MUC]-1, MUC-2 and MUC-4) and the antimicrobial peptide Reg3β were increased after probiotic supplementation.CONCLUSIONBased on these results, the new strain of P. pentosaceus alleviated ethanol-induced liver injury by reversing gut microbiota dysbiosis, regulating intestinal SCFA metabolism, improving intestinal barrier function, and reducing circulating levels of endotoxin and proinflammatory cytokines and chemokines. Thus, this strain is a potential probiotic treatment for ALD.

Sexually Dimorphic Responses to a High-Refined Carbohydrate Diet in a Nonalcoholic Fatty Liver Disease Mouse Model

Abstract Objectives Nonalcoholic fatty liver disease (NAFLD) incidence and prevalence have been reported to be higher in men than women, however, the effects of sexual dimorphism on NAFLD risk and progression have not been adequately examined. Our lab has previously shown that a liquid high-refined carbohydrate diet (HRCD) induced more severe hepatic steatosis compared to an isocaloric high fat diet in male mice. Also, HRCD-induced reduction in sirtuin 1 (SIRT1), an NAD-dependent deacetylase protein, has previously been implicated in NAFLD pathogenesis. Therefore, we investigated whether there were sexually dimorphic responses to a liquid high-refined carbohydrate diet (HRCD) in male and female, wildtype and SIRT1-deficient mice. Methods Male and female 10–12-week-old wildtype (SIRT1 +/+: n = 12; M = 6, F = 6) and mice carrying a heterozygous H355Y SIRT1 point mutation (SIRT1 +/y: n = 14; M = 7, F = 7) were both fed a HRCD (Lieber-DeCarli liquid diet supplemented with maltose dextrin; 47% energy from refined carbohydrate, Dyets, #710,260) for 5 weeks and 9 weeks. Hepatic gene expression was examined using qRT-PCR. Plasma ALT (alanine transaminase) and hepatic MDA (malondialdehyde) levels were determined using colorimetric assay kits. Hepatic steatosis scoring was conducted by analyzing Hematoxylin and Eosin (H&amp;E) stains. Results 9 weeks of HRCD induced significantly less hepatic steatosis in female mice irrespective of genotype compared to male mice as determined by grading of H&amp;E stains (P &amp;lt; 0.05). Furthermore, liver expression of several fatty acid oxidation genes (CPT1, ACOX1) was significantly higher in females (P &amp;lt; 0.05), which potentially suggests increased fatty acid oxidation. Additionally, female mice had significantly increased antioxidant gene expression (GPX4, SOD1, SOD2, Catalase) and significantly lower hepatic MDA (P &amp;lt; 0.05), which indicate an increased capacity to mitigate oxidative stress. Lastly, plasma ALT levels were significantly lower in females compared to males after 9 weeks of HRCD (P &amp;lt; 0.05). Conclusions Collectively, these data indicate that female mice are moderately protected against HRCD-induced NAFLD compared to male mice, potentially through increased hepatic fatty acid oxidation and superior mitigation of oxidative stress due to increased antioxidant system gene expression in the liver. Funding Sources HNRCA, USDA/ARS Grants.

Effects of Moderate Ethanol Consumption with a n‐3 Diet on Lipid Profile and Liver Function in Mice

Several studies have found a non‐linear relationship between alcohol consumption and cardiovascular disease (CVD) risk, suggesting that low or moderate consumption of alcohol may provide cardioprotective effects, while excessive alcohol consumption increases the risk of CVD. Studies also suggest that increased dietary consumption of n‐3 fatty acids may reduce CVD risk. The purpose of this study is to examine the effects of moderate ethanol consumption as a function of dietary n‐6:n‐3 fatty acid composition on markers associated with cardiovascular disease and liver dysfunction in mice. Twenty‐three mice (12 male, 11 female) consumed an 18% ethanol solution or 26.9% maltose dextrin solution (non‐ethanol isocaloric control) for 12 weeks. In each group, half of the mice were fed either a high n‐6 (n‐6:n‐3 = 50:1) diet or a balanced n‐3 (n‐6:n‐3 = 1:1) diet ad libitum. There were no differences in initial body weight between the two groups; however, the control group gained significantly more weight than the ethanol group (P = 0.020) associated with higher maltose dextrin fluid intake (P &lt; 0.001). In the control group, the balanced n‐3 diet resulted in significantly lower aspartate aminotransferase (AST) levels (P = 0.010) compared to the high n‐6 diet. In the ethanol group, the balanced n‐3 diet resulted in lower levels of serum triglycerides (P = 0.086), total cholesterol (P= 0.096), and low‐density lipoprotein (LDL) levels (P = 0.053) that were approaching significance. These findings indicate that a diet with a balanced n‐6 to n‐3 ratio may improve several lipid profile markers associated with CVD in mice who concurrently consume moderate amounts of alcohol.Support or Funding InformationNIH AA023291

Intestinal MiR194 Regulates Hepatic Bile Acids through Intestinal FXR‐FGF15 Pathway in Alcoholic Liver Disease in Mice

BackgroundAlcoholic liver disease (ALD) is characterized by gut dysbiosis, higher hepatic and circulating levels of bile acids (BAs) and intestinal epithelial dysfunction. BA homeostasis is modulated by hepatic and intestinal nuclear farnesoid X receptor (FXR, Nr1h4), a ligand activated transcription factor, through regulating key genes involved in bile acid synthesis and metabolism. BA synthesis regulation by intestinal FXR is through induction of fibroblast growth factor 15 (FGF15) that inhibits hepatic Cyp7a1, the key gene in BA de novo synthesis. Previous studies showed that FXR expression is suppressed miR194 in various tissues. In this study, we investigated the regulation of FXR by miR194 in intestinal tissues in mice with ALD.MethodsC57BL6 and intestinal specific FXR knockout (FXRΔIEC) mice were subjected to the Lieber DeCarli liquid diet containing 5% alcohol for 10 days, and one bonus of alcohol (5 g/kg) was gavaged on last day 9 hours before tissue collecting. Control mice were fed isocaloric maltose dextrin. A group of mice was supplemented with Lactobacillus rhamnosus GG (LGG) at a dose of 109 CFU/day for 10 days before alcohol exposure.ResultsAlcohol feeding significantly increased hepatic fat accumulation and serum ALT and AST levels. Interestingly, the alcohol‐induced effects were associated with an increased expression of miR194 and a decreased expression of FXR and FGF15 in the intestine, and with an upregulation of hepatic Cyp7a1 and BA concentration. When the mice were supplemented with probiotic LGG, intestinal miR194 expression was suppressed, FXR‐FGF15 signaling pathway was upregulated, and hepatic BA levels were decreased. Alcohol induced hepatic steatosis and injury were markedly prevented by LGG. More importantly, the beneficial effects of LGG on hepatic BA homeostasis in ALD mice were diminished when the intestinal epithelial FXR was deleted. In vitro study showed that alcohol treatment increased miR194 and decreased FXR mRNA, and inhibition of miR194 attenuated the suppression of FXR mRNA by alcohol.ConclusionsTaken together, our results demonstrate that intestinal FXR‐FGF15 signaling plays a pivotal role in maintaining hepatic BA homeostasis in ALD mice and probiotic LGG protects against ALD through maintaining BA homeostasis regulated by intestinal miR194‐FXR‐FGF15‐mediated signaling pathways.Support or Funding InformationSupported by grants from NIH.

Critical review on conventional spectroscopic α-amylase activity detection methods: merits, demerits, and future prospects.

α-Amylase is an endoenzyme that catalyses the hydrolysis of internal α-l,4 glycosidic linkages in polysaccharides to produce maltose, maltotriose, and α-limit dextrins. It is widely used in the laboratorial and industrial workflow for several applications. There are several methods utilizing different techniques and substrates to assess α-amylase activity, among which the spectroscopic methods have found widespread applicability due to their ease of use and cost-effectiveness. Depending upon the reaction principle, these assays are classified into four groups: reducing sugar, enzymatic, chromogenic, and amyloclastic methods. Despite the presence of numerous methods, there is no general reliable method to assess α-amylase activity. Each method is shown to have its own merits and demerits. Many improvements have been made to make the available methods more accurate, reliable, and easy. This communication briefly discusses the basic reaction mechanisms and critically reviews the advantages and shortcomings associated with each method. Further recommendations are made for future development. © 2020 Society of Chemical Industry.

Distribution of Phosphatidylethanol in Maternal and Fetal Compartments After Chronic Gestational Binge Alcohol Exposure.

Phosphatidylethanol (PEth) is a promising biomarker for gestational alcohol exposure. Studies show PEth accumulation in maternal and fetal blood following alcohol exposure; however, distribution of specific PEth homologues (16:0/18:1, 16:0/18:2, 16:0/20:4) in maternal and fetal blood is unknown. Additionally, PEth levels in highly vulnerable FASD targets in maternal and fetal compartments remain unexplored. We hypothesized that all 3 major PEth homologues will be detectable in the maternal and fetal blood, the maternal uterine artery (a reproductive tissue that delivers oxygen and nutrients to fetoplacental unit), and fetal brain regions following gestational binge alcohol exposure and that homologue distribution profiles will be tissue-specific. Pregnant rats received once-daily orogastric gavage of alcohol (Alcohol; BAC 216mg/dl@4.5g/kg/d; BAC 289mg/dl@6g/kg/d) or iso-caloric maltose dextrin (Pair-fed control) from gestation days (GD) 5 to 20 or 21. Following chronic exposure, maternal and fetal tissues were analyzed for PEth homologue concentrations utilizing LC-MS/MS technology. All 3 PEth homologues were detected in alcohol-exposed maternal blood, fetal blood, maternal uterine artery, and fetal brain regions (hippocampus, cerebral cortex, and cerebellum). In both maternal and fetal blood, respectively, PEth 16:0/18:2 was more abundant compared to 16:0/18:1 (p<0.0001,~66%,↑; p=0.0159, 20.4%↑) and 16:0/20:4 (p=0.0072,~25%↑; p=0.0187, 19.4%↑). Maternal PEth 16:0/20:4 was~42% higher than 16:0/18:1 (p=0.0015). Maternal PEth 16:0/18:2 and 16:0/20:4 were~25%↑ and~20%↑ higher than in fetal blood (p<0.05). No homologue differences were detected in the maternal uterine artery. In all fetal brain regions, PEth 16:0/18:1 was significantly higher (p<0.0001) than 16:0/18:2 (~48 to 78%↑) and 16:0/20:4 (~31 to 62%↑) concentrations. PEth 16:0/20:4 was~18% higher than 16:0/18:1 (p<0.05) in the fetal hippocampus and cortex. All major PEth homologues were detected in maternal and fetal blood following chronic gestational binge alcohol exposure; homologue distribution profiles were tissue-specific. This study also provides insights into PEth accumulation in critical FASD targets, specifically the maternal uterine artery and fetal brain.

Voluntary Chronic Heavy Alcohol Consumption in Male Rhesus Macaques Suppresses Cancellous Bone Formation and Increases Bone Marrow Adiposity.

Chronic heavy alcohol consumption is an established risk factor for bone fracture, but comorbidities associated with alcohol intake may contribute to increased fracture rates in alcohol abusers. To address the specific effects of alcohol on bone, we used a nonhuman primate model and evaluated voluntary alcohol consumption on: (i) global markers of bone turnover in blood and (ii) cancellous bone mass, density, microarchitecture, turnover, and microdamage in lumbar vertebra. Following a 4-month induction period, 6-year-old male rhesus macaques (Macaca mulatta, n=13) voluntarily self-administered water or ethanol (EtOH; 4% w/v) for 22h/d, 7 d/wk, for a total of 12months. Control animals (n=9) consumed an isocaloric maltose-dextrin solution. Tetracycline hydrochloride was administered orally 17 and 3days prior to sacrifice to label mineralizing bone surfaces. Global skeletal response to EtOH was evaluated by measuring plasma osteocalcin and carboxyterminal collagen cross-links (CTX). Local response was evaluated in lumbar vertebra using dual-energy X-ray absorptiometry, microcomputed tomography, static and dynamic histomorphometry, and histological assessment of microdamage. Monkeys in the EtOH group consumed an average of 2.8±0.2 (mean±SE) g/kg/d of EtOH (30±2% of total calories), resulting in an average blood EtOH concentration of 88.3±8.8mg/dl 7hours after the session onset. Plasma CTX and osteocalcin tended to be lower in EtOH-consuming monkeys compared to controls. Significant differences in bone mineral density in lumbar vertebrae 1 to 4 were not detected with treatment. However, cancellous bone volume fraction (in cores biopsied from the central region of the third vertebral body) was lower in EtOH-consuming monkeys compared to controls. Furthermore, EtOH-consuming monkeys had lower osteoblast perimeter and mineralizing perimeter, no significant difference in osteoclast perimeter, and higher bone marrow adiposity than controls. No significant differences between groups were detected in microcrack density (2nd lumbar vertebra). Voluntary chronic heavy EtOH consumption reduces cancellous bone formation in lumbar vertebra by decreasing osteoblast-lined bone perimeter, a response associated with an increase in bone marrow adiposity.

Hippocampal transcriptome reveals novel targets of FASD pathogenesis.

IntroductionPrenatal alcohol exposure can contribute to fetal alcohol spectrum disorders (FASD), characterized by a myriad of developmental impairments affecting behavior and cognition. Studies show that many of these functional impairments are associated with the hippocampus, a structure exhibiting exquisite vulnerability to developmental alcohol exposure and critically implicated in learning and memory; however, mechanisms underlying alcohol‐induced hippocampal deficits remain poorly understood. By utilizing a high‐throughput RNA‐sequencing (RNA‐seq) approach to address the neurobiological and molecular basis of prenatal alcohol‐induced hippocampal functional deficits, we hypothesized that chronic binge prenatal alcohol exposure alters gene expression and global molecular pathways in the fetal hippocampus.MethodsTimed‐pregnant Sprague–Dawley rats were randomly assigned to a pair‐fed control (PF) or binge alcohol (ALC) treatment group on gestational day (GD) 4. ALC dams acclimatized from GDs 5–10 with a daily treatment of 4.5 g/kg alcohol and subsequently received 6 g/kg on GDs 11–20. PF dams received a once daily maltose dextrin gavage on GDs 5–20, isocalorically matching ALC counterparts. On GD 21, bilateral hippocampi were dissected, flash frozen, and stored at −80°C. Total RNA was then isolated from homogenized tissues. Samples were normalized to ~4nM and pooled equally. Sequencing was performed by Illumina NextSeq 500 on a 75 cycle, single‐end sequencing run.ResultsRNA‐seq identified 13,388 genes, of these, 76 genes showed a significant difference (p < 0.05, log2 fold change ≥2) in expression between the PF and ALC groups. Forty‐nine genes showed sex‐dependent dysregulation; IPA analysis showed among female offspring, dysregulated pathways included proline and citrulline biosynthesis, whereas in males, xenobiotic metabolism signaling and alaninine biosynthesis etc. were altered.ConclusionWe conclude that chronic binge alcohol exposure during pregnancy dysregulates fetal hippocampal gene expression in a sex‐specific manner. Identification of subtle, transcriptome‐level dysregulation in hippocampal molecular pathways offers potential mechanistic insights underlying FASD pathogenesis.

Intestinal HIF‐1α Deletion Exacerbates Alcoholic Liver Disease through Inducing Intestinal Dysbiosis and Barrier Dysfunction

ObjectivesAlcoholic liver disease (ALD) is characterized by gut dysbiosis and increased gut permeability. Hypoxia Induced Factor 1α (HIF‐1α) has been implicated in transcriptional regulation of intestinal barrier integrity and inflammation. We aimed to test the hypothesis that HIF‐1α plays a critical role in gut microbiota homeostasis and the maintenance of intestinal barrier integrity in a mouse model of ALD.MethodsWide type (WT) and intestinal epithelial‐specific HIF‐1α knockout mice (IEhif‐1α−/−) were pair‐fed modified Lieber‐DeCarli liquid diet containing 5% (w/v) alcohol or isocaloric maltose dextrin for 24 days. Serum levels of ALT and endotoxin were determined. Fecal microbiota were assessed. Liver steatosis and injury, and intestinal barrier integrity were evaluated.ResultsAlcohol feeding increased serum levels of ALT and LPS, hepatic triglyceride concentration, and liver injury in the WT mice. These deleterious effects were exaggerated in IEhif‐1α−/− mice. Alcohol exposure resulted in greater reduction of the expression of intestinal epithelial tight junction proteins, claudin‐1 and occludin, in IEhif‐1α−/− mice. In addition, cathelicidin‐related antimicrobial peptide (CRAMP) and intestinal trefoil factor (ITF) were further decreased by alcohol in IEhif‐1α−/− mice. Metagenomic analysis showed an increased gut dysbiosis with a significantly decreased firmicutes/bacteroidetes ratio in IEhif‐1α−/− mice compared to the WT mice exposed to alcohol. An increased abundance of Akkermansia and a decreased level of Lactobacillus in IEhif‐1α−/− mice were observed. Non‐absorbable antibiotics treatment reversed the liver steatosis in both WT and IEhif‐1α−/− mice.ConclusionIntestinal HIF‐1α is essential for the adaptation response to alcohol exposure‐induced changes in intestinal microbiota and barrier function associated with elevated endotoxemia and hepatic steatosis and injury.Support or Funding InformationNIHThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Detecting Neurodevelopmental Effects of Early-Gestation Ethanol Exposure: A Nonhuman Primate Model of Ethanol Drinking During Pregnancy.

Gestational ethanol (EtOH) exposure is associated with multiple developmental abnormalities, collectively termed fetal alcohol spectrum disorder (FASD). While the majority of women abstain from EtOH following knowledge of pregnancy, one contributing factor to the high FASD prevalence is that pregnancy is not detected until 4 to 6weeks. Thus, EtOH consumption continues during the initial stages of fetal development. An experimental protocol is described in which rhesus macaques self-administer 1.5g/kg/d EtOH (or isocaloric maltose dextrin) prior to pregnancy and through the first 60days of a 168-day gestation term. Menstrual cycles were monitored, including measurements of circulating estradiol and progesterone levels. The latency to consume 1.5g/kg EtOH and blood EtOH concentration (BEC) was measured. Twenty-eight fetuses (14 EtOH and 14 controls) were generated in this study. EtOH did not affect menstrual cycles or the probability of successful breeding. No EtOH-induced gross adverse effects on pregnancy were observed. Individual variability in latency to complete drinking translated into variability in BEC, measured 90minutes following session start. Drinking latencies in controls and EtOH drinkers were longer in the second gestational month than in the first. All pregnancies reached the planned experimental time point of G85, G110, or G135, when in utero MRIs were performed, fetuses were delivered by caesarean section, and brains were evaluated with exvivo procedures, including slice electrophysiology. Fetal tissues have been deposited to the Monkey Alcohol Tissue Research Resource. This FASD model takes advantage of the similarities between humans and rhesus macaques in gestational length relative to brain development, as well as similarities in EtOH self-administration and metabolism. The daily 1.5g/kg dose of EtOH through the first trimester does not influence pregnancy success rates. However, pregnancy influences drinking behavior during the second month of pregnancy. Future publications using this model will describe the effect of early-gestation EtOH exposure on anatomical and functional brain development at subsequent gestational ages.