1   871 134 CHRONIC ALCOHOL EXPOSURE AFFECTS PANCREATIC ACINAR MITOCHONDRIAL THIAMIN PYROPHOSPHATE UPTAKE: STUDIES WITH MOUSE 266-6 CELL LINE AND PRIMARY CELLS. THIAMIN IS ESSENTIAL FOR NORMAL METABOLIC ACTIVITY OF ALL MAMMALIAN CELLS, INCLUDING THOSE OF THE PANCREAS. CELLS OBTAIN THIAMIN FROM THEIR SURROUNDINGS AND ENZYMATICALLY CONVERT IT INTO THIAMIN PYROPHOSPHATE (TPP) IN THE CYTOPLASM; TPP IS THEN TAKEN UP BY MITOCHONDRIA VIA A SPECIFIC CARRIER THE MITOCHONDRIAL TPP TRANSPORTER (MTPPT; PRODUCT OF THE SLC25A19 GENE). CHRONIC ALCOHOL EXPOSURE NEGATIVELY IMPACTS THE HEALTH OF PANCREATIC ACINAR CELLS (PAC), BUT ITS EFFECT ON PHYSIOLOGICAL/MOLECULAR PARAMETERS OF MTPPT IS NOT KNOWN. WE ADDRESSED THIS ISSUE USING MOUSE PANCREATIC ACINAR TUMOR CELL LINE 266-6 AND PRIMARY PAC OF WILD-TYPE AND TRANSGENIC MICE CARRYING THE SLC25A19 PROMOTER THAT WERE FED ALCOHOL CHRONICALLY. CHRONIC ALCOHOL EXPOSURE OF 266-6 CELLS (BUT NOT TO ITS NONOXIDATIVE METABOLITES ETHYL PALMITATE AND ETHYL OLEATE) LED TO A SIGNIFICANT INHIBITION IN MITOCHONDRIAL TPP UPTAKE, WHICH WAS ASSOCIATED WITH A DECREASED EXPRESSION OF MTPPT PROTEIN, MRNA, AND ACTIVITY OF THE SLC25A19 PROMOTER. SIMILARLY, CHRONIC ALCOHOL FEEDING OF MICE LED TO A SIGNIFICANT INHIBITION IN EXPRESSION OF MTPPT PROTEIN, MRNA, HETEROGENEOUS NUCLEAR RNA, AS WELL AS IN ACTIVITY OF SLC25A19 PROMOTER IN PAC. WHILE CHRONIC ALCOHOL EXPOSURE DID NOT AFFECT DNA METHYLATION OF THE SLC25A19 PROMOTER, A SIGNIFICANT DECREASE IN HISTONE H3 EUCHROMATIN MARKERS AND AN INCREASE IN H3 HETEROCHROMATIN MARKER WERE OBSERVED. THESE FINDINGS SHOW, FOR THE FIRST TIME, THAT CHRONIC ALCOHOL EXPOSURE NEGATIVELY IMPACTS PANCREATIC MTPPT, AND THAT THIS EFFECT IS EXERTED, AT LEAST IN PART, AT THE LEVEL OF SLC25A19 TRANSCRIPTION AND APPEARS TO INVOLVE EPIGENETIC MECHANISM(S).	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
2  1787  57 EFFECT OF CHRONIC ALCOHOL EXPOSURE ON GUT VITAMIN B7 UPTAKE: INVOLVEMENT OF EPIGENETIC MECHANISMS AND EFFECT OF ALCOHOL METABOLITES. VITAMIN B7 (BIOTIN) IS ESSENTIAL FOR NORMAL HEALTH AND ITS DEFICIENCY/SUBOPTIMAL LEVELS OCCUR IN A VARIETY OF CONDITIONS INCLUDING CHRONIC ALCOHOLISM. MAMMALS, INCLUDING HUMANS, OBTAIN BIOTIN FROM DIET AND GUT-MICROBIOTA VIA ABSORPTION ALONG THE INTESTINAL TRACT. THE ABSORPTION PROCESS IS CARRIER MEDIATED AND INVOLVES THE SODIUM-DEPENDENT MULTIVITAMIN TRANSPORTER (SMVT; SLC5A6). WE HAVE PREVIOUSLY SHOWN THAT CHRONIC ALCOHOL EXPOSURE SIGNIFICANTLY INHIBITS INTESTINAL/COLONIC BIOTIN UPTAKE VIA SUPPRESSION OF SLC5A6 TRANSCRIPTION IN ANIMAL AND CELL LINE MODELS. HOWEVER, LITTLE IS KNOWN ABOUT THE TRANSCRIPTIONAL/EPIGENETIC FACTORS THAT MEDIATE THIS SUPPRESSION. IN ADDITION, THE EFFECT OF ALCOHOL METABOLITES (GENERATED VIA ALCOHOL METABOLISM BY GUT MICROBIOTA AND HOST TISSUES) ON BIOTIN UPTAKE IS STILL UNKNOWN. TO ADDRESS THESE QUESTIONS, WE FIRST DEMONSTRATED THAT CHRONIC ALCOHOL EXPOSURE INHIBITS SMALL INTESTINAL AND COLONIC BIOTIN UPTAKE AND SMVT EXPRESSION IN HUMAN DIFFERENTIATED ENTEROID AND COLONOID MONOLAYERS. WE THEN SHOWED THAT CHRONIC ALCOHOL EXPOSURES OF BOTH, CACO-2 CELLS AND MICE, ARE ASSOCIATED WITH A SIGNIFICANT SUPPRESSION IN EXPRESSION OF THE NUCLEAR FACTOR KLF-4 (NEEDED FOR SLC5A6 PROMOTER ACTIVITY), AS WELL AS WITH EPIGENETIC ALTERATIONS (HISTONE MODIFICATIONS). WE ALSO FOUND THAT CHRONIC EXPOSURE OF NCM460 HUMAN COLONIC EPITHELIAL CELLS AS WELL AS HUMAN DIFFERENTIATED COLONOID MONOLAYERS, TO ALCOHOL METABOLITES (ACETALDEHYDE, ETHYL PALMITATE, ETHYL OLEATE) SIGNIFICANTLY INHIBITED BIOTIN UPTAKE AND SMVT EXPRESSION. THESE FINDINGS SHED LIGHT ONTO THE MOLECULAR/EPIGENETIC MECHANISMS THAT MEDIATE THE INHIBITORY EFFECT OF CHRONIC ALCOHOL EXPOSURE ON INTESTINAL BIOTIN UPTAKE. THEY FURTHER SHOW THAT ALCOHOL METABOLITES ARE ALSO CAPABLE OF INHIBITING BIOTIN UPTAKE IN THE GUT.NEW & NOTEWORTHY USING COMPLEMENTARY MODELS, INCLUDING HUMAN DIFFERENTIATED ENTEROID AND COLONOID MONOLAYERS, THIS STUDY SHOWS THE INVOLVEMENT OF MOLECULAR AND EPIGENETIC MECHANISMS IN MEDIATING THE INHIBITORY EFFECT OF CHRONIC ALCOHOL EXPOSURE ON BIOTIN UPTAKE ALONG THE INTESTINAL TRACT. THE STUDY ALSO SHOWS THAT ALCOHOL METABOLITES (GENERATED BY GUT MICROBIOTA AND HOST TISSUES) CAUSE INHIBITION IN GUT BIOTIN UPTAKE.	2021	
                                                                                                                                                                                                                    
3   873  75 CHRONIC ALCOHOL EXPOSURE INHIBITS BIOTIN UPTAKE BY PANCREATIC ACINAR CELLS: POSSIBLE INVOLVEMENT OF EPIGENETIC MECHANISMS. CHRONIC EXPOSURE TO ALCOHOL AFFECTS DIFFERENT PHYSIOLOGICAL ASPECTS OF PANCREATIC ACINAR CELLS (PAC), BUT ITS EFFECT ON THE UPTAKE PROCESS OF BIOTIN IS NOT KNOWN. WE ADDRESSED THIS ISSUE USING MOUSE-DERIVED PANCREATIC ACINAR 266-6 CELLS CHRONICALLY EXPOSED TO ALCOHOL AND WILD-TYPE AND TRANSGENIC MICE (CARRYING THE HUMAN SLC5A6 5'-PROMOTER) FED ALCOHOL CHRONICALLY. FIRST WE ESTABLISHED THAT BIOTIN UPTAKE BY PAC IS NA(+) DEPENDENT AND CARRIER MEDIATED AND INVOLVES SODIUM-DEPENDENT MULTIVITAMIN TRANSPORTER (SMVT). CHRONIC EXPOSURE OF 266-6 CELLS TO ALCOHOL LED TO A SIGNIFICANT INHIBITION IN BIOTIN UPTAKE, EXPRESSION OF SMVT PROTEIN, AND MRNA AS WELL AS IN THE ACTIVITY OF THE SLC5A6 PROMOTER. SIMILARLY, CHRONIC ALCOHOL FEEDING OF WILD-TYPE AND TRANSGENIC MICE CARRYING THE SLC5A6 PROMOTER LED TO A SIGNIFICANT INHIBITION IN BIOTIN UPTAKE BY PAC, AS WELL AS IN THE EXPRESSION OF SMVT PROTEIN AND MRNA AND THE ACTIVITY OF THE SLC5A6 PROMOTERS EXPRESSED IN THE TRANSGENIC MICE. WE ALSO FOUND THAT CHRONIC ALCOHOL FEEDING OF MICE IS ASSOCIATED WITH A SIGNIFICANT INCREASE IN THE METHYLATION STATUS OF CPG ISLANDS PREDICTED TO BE IN THE MOUSE SLC5A6 PROMOTERS AND A DECREASE IN THE LEVEL OF EXPRESSION OF TRANSCRIPTION FACTOR KLF-4, WHICH PLAYS AN IMPORTANT ROLE IN REGULATING SLC5A6 PROMOTER ACTIVITY. THESE RESULTS DEMONSTRATE, FOR THE FIRST TIME, THAT CHRONIC ALCOHOL EXPOSURE NEGATIVELY IMPACTS BIOTIN UPTAKE IN PAC AND THAT THIS EFFECT IS EXERTED (AT LEAST IN PART) AT THE LEVEL OF TRANSCRIPTION OF THE SLC5A6 GENE AND MAY INVOLVE EPIGENETIC/MOLECULAR MECHANISMS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
4  3727  77 INHIBITION OF PANCREATIC ACINAR MITOCHONDRIAL THIAMIN PYROPHOSPHATE UPTAKE BY THE CIGARETTE SMOKE COMPONENT 4-(METHYLNITROSAMINO)-1-(3-PYRIDYL)-1-BUTANONE. THIAMIN IS ESSENTIAL FOR NORMAL METABOLISM IN PANCREATIC ACINAR CELLS (PAC) AND IS OBTAINED FROM THEIR MICROENVIRONMENT THROUGH SPECIFIC PLASMA-MEMBRANE TRANSPORTERS, CONVERTED TO THIAMIN PYROPHOSPHATE (TPP) IN THE CYTOPLASM, FOLLOWED BY UPTAKE OF TPP BY MITOCHONDRIA THROUGH THE MITOCHONDRIAL TPP (MTPP) TRANSPORTER (MTPPT; PRODUCT OF SLC25A19 GENE). TPP IS ESSENTIAL FOR NORMAL MITOCHONDRIAL FUNCTION. WE EXAMINED THE EFFECT OF LONG-TERM/CHRONIC EXPOSURE OF PAC IN VITRO (PANCREATIC ACINAR 266-6 CELLS) AND IN VIVO (WILD-TYPE OR TRANSGENIC MICE CARRYING THE SLC25A19 PROMOTER) OF THE CIGARETTE SMOKE TOXIN, 4-(METHYLNITROSAMINO)-1-(3-PYRIDYL)-1-BUTANONE (NNK), ON THE MTPP UPTAKE PROCESS. OUR IN VITRO AND IN VIVO FINDINGS DEMONSTRATE THAT NNK NEGATIVELY AFFECTS MTPP UPTAKE AND REDUCED EXPRESSION OF MTPPT PROTEIN, MTPPT MRNA, AND HETEROGENOUS NUCLEAR RNA, AS WELL AS SLC25A19 PROMOTER ACTIVITY. THE EFFECT OF NNK ON SLC25A19 TRANSCRIPTION WAS NEITHER MEDIATED BY CHANGES IN EXPRESSION OF TRANSCRIPTIONAL FACTOR NFY-1 (KNOWN TO DRIVE SLC25A19 TRANSCRIPTION), NOR DUE TO CHANGES IN METHYLATION PROFILE OF THE SLC25A19 PROMOTER. RATHER, IT APPEARS TO BE DUE TO CHANGES IN HISTONE MODIFICATIONS THAT INVOLVE SIGNIFICANT DECREASES IN HISTONE H3K4-TRIMETHYLATION AND H3K9-ACETYLATION (ACTIVATION MARKERS). THE EFFECT OF NNK ON MTPPT FUNCTION IS MEDIATED THROUGH THE NONNEURONAL ALPHA7-NICOTINIC ACETYLCHOLINE RECEPTOR (ALPHA7-NACHR), AS INDICATED BY BOTH IN VITRO (USING THE NACHR ANTAGONIST MECAMYLAMINE) AND IN VIVO (USING AN ALPHA7-NACHR(-/-) MOUSE MODEL) STUDIES. THESE FINDINGS DEMONSTRATE THAT CHRONIC EXPOSURE OF PAC TO NNK NEGATIVELY IMPACTS PAC MTPP UPTAKE. THIS EFFECT APPEARS TO BE EXERTED AT THE LEVEL OF SLC25A19 TRANSCRIPTION, INVOLVE EPIGENETIC MECHANISM(S), AND IS MEDIATED THROUGH THE ALPHA7-NACHR.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
5  3556  28 IMPACT OF ALCOHOL CONSUMPTION ON MALE FERTILITY POTENTIAL: A NARRATIVE REVIEW. ALCOHOL ABUSE DISORDER IS A SERIOUS CONDITION, IMPLICATING MORE THAN 15 MILLION PEOPLE AGED 12 YEARS AND OLDER IN 2019 IN THE UNITED STATES. ETHANOL (OR ETHYL ALCOHOL) IS MAINLY OXIDIZED IN THE LIVER, RESULTING IN THE SYNTHESIS OF ACETALDEHYDE AND ACETATE, WHICH ARE TOXIC AND CARCINOGENIC METABOLITES, AS WELL AS IN THE GENERATION OF A REDUCTIVE CELLULAR ENVIRONMENT. MOREOVER, ETHANOL CAN INTERACT WITH LIPIDS, GENERATING FATTY ACID ETHYL ESTERS AND PHOSPHATIDYLETHANOL, WHICH INTERFERE WITH PHYSIOLOGICAL CELLULAR PATHWAYS. THIS NARRATIVE REVIEW SUMMARIZES THE IMPACT OF EXCESSIVE ALCOHOL CONSUMPTION ON MALE FERTILITY BY DESCRIBING ITS METABOLISM AND HOW ETHANOL CONSUMPTION MAY INDUCE CELLULAR DAMAGE. FURTHERMORE, THE IMPACT OF ALCOHOL CONSUMPTION ON HORMONAL REGULATION, SEMEN QUALITY, AND GENETIC AND EPIGENETIC REGULATIONS IS DISCUSSED BASED ON EVIDENCE FROM ANIMAL AND HUMAN STUDIES, FOCUSING ON THE CONSEQUENCES ON THE OFFSPRING. FINALLY, THE LIMITATIONS OF THE CURRENT EVIDENCE ARE DISCUSSED. OUR REVIEW HIGHLIGHTS THE ASSOCIATION BETWEEN CHRONIC ALCOHOL CONSUMPTION AND POOR SEMEN QUALITY, MAINLY DUE TO THE DEVELOPMENT OF OXIDATIVE STRESS, AS WELL AS ITS GENOTOXIC IMPACT ON HORMONAL REGULATION AND DNA INTEGRITY, AFFECTING THE OFFSPRING'S HEALTH. NEW LANDSCAPES OF INVESTIGATION ARE PROPOSED FOR THE IDENTIFICATION OF MOLECULAR MARKERS FOR ALCOHOL-ASSOCIATED INFERTILITY, WITH A FOCUS ON ADVANCED OMICS-BASED APPROACHES APPLIED TO THE ANALYSIS OF SEMEN SAMPLES.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
6  6666  72 UPTAKE OF ASCORBIC ACID BY PANCREATIC ACINAR CELLS IS NEGATIVELY IMPACTED BY CHRONIC ALCOHOL EXPOSURE. VITAMIN C (ASCORBIC ACID, AA) IS INDISPENSABLE FOR NORMAL METABOLISM OF ALL MAMMALIAN CELLS INCLUDING PANCREATIC ACINAR CELLS (PACS). PACS OBTAIN AA FROM THEIR SURROUNDINGS VIA TRANSPORT ACROSS THE CELL MEMBRANE. CHRONIC ALCOHOL EXPOSURE NEGATIVELY AFFECTS BODY AA HOMEOSTASIS; IT ALSO INHIBITS UPTAKE OF OTHER MICRONUTRIENTS INTO PACS, BUT ITS EFFECT ON AA UPTAKE IS NOT CLEAR. WE EXAMINED THIS ISSUE USING BOTH IN VITRO (266-6 CELLS) AND IN VIVO (MICE) MODELS OF CHRONIC ALCOHOL EXPOSURE. FIRST, WE DETERMINED THE RELATIVE EXPRESSION OF THE AA TRANSPORTERS 1 AND 2 [I.E., SODIUM-DEPENDENT VITAMIN C TRANSPORTER-1 (SVCT-1) AND SVCT-2] IN MOUSE AND HUMAN PACS AND FOUND SVCT-2 TO BE THE PREDOMINANT TRANSPORTER. CHRONIC EXPOSURE OF 266-6 CELLS TO ALCOHOL SIGNIFICANTLY INHIBITED AA UPTAKE AND CAUSED A MARKED REDUCTION IN SVCT-2 EXPRESSION AT THE PROTEIN, MRNA, AND HETEROGENEOUS NUCLEAR RNA (HNRNA) LEVELS. SIMILARLY, CHRONIC ALCOHOL FEEDING OF MICE SIGNIFICANTLY INHIBITED AA UPTAKE AND CAUSED A MARKED REDUCTION IN LEVEL OF EXPRESSION OF THE SVCT-2 PROTEIN, MRNA, AND HNRNA. THESE FINDINGS SUGGEST POSSIBLE INVOLVEMENT OF TRANSCRIPTIONAL MECHANISM(S) IN MEDIATING CHRONIC ALCOHOL EFFECT ON AA UPTAKE BY PACS. WE ALSO OBSERVED SIGNIFICANT EPIGENETIC CHANGES (HISTONE MODIFICATIONS) IN THE SLC23A2 GENE (REDUCTION IN H3K4ME3 LEVEL AND AN INCREASE IN H3K27ME3 LEVEL) IN THE ALCOHOL-EXPOSED 266-6 CELLS. THESE FINDINGS SHOW THAT CHRONIC ALCOHOL EXPOSURE INHIBITS PAC AA UPTAKE AND THAT THE EFFECT IS MEDIATED, IN PART, AT THE LEVEL OF TRANSCRIPTION OF THE SLC23A2 GENE AND MAY INVOLVE EPIGENETIC MECHANISM(S).	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
7  6086  39 THE EFFECTS OF ACETALDEHYDE EXPOSURE ON HISTONE MODIFICATIONS AND CHROMATIN STRUCTURE IN HUMAN LUNG BRONCHIAL EPITHELIAL CELLS. AS THE PRIMARY METABOLITE OF ALCOHOL AND THE MOST ABUNDANT CARCINOGEN IN TOBACCO SMOKE, ACETALDEHYDE IS LINKED TO A NUMBER OF HUMAN DISEASES ASSOCIATED WITH CHRONIC ALCOHOL CONSUMPTION AND SMOKING INCLUDING CANCERS. IN ADDITION TO DIRECT DNA DAMAGE AS A RESULT OF THE FORMATION OF ACETALDEHYDE-DNA ADDUCTS, ACETALDEHYDE MAY ALSO INDIRECTLY IMPACT PROPER GENOME FUNCTION THROUGH THE FORMATION OF PROTEIN ADDUCTS. HISTONE PROTEINS ARE THE MAJOR COMPONENT OF THE CHROMATIN. POST-TRANSLATIONAL HISTONE MODIFICATIONS (PTMS) ARE CRITICALLY IMPORTANT FOR THE MAINTENANCE OF GENETIC AND EPIGENETIC STABILITY. HOWEVER, LITTLE IS KNOWN ABOUT HOW ACETALDEHYDE-HISTONE ADDUCTS AFFECT HISTONE MODIFICATIONS AND CHROMATIN STRUCTURE. THE RESULTS OF PROTEIN CARBONYL ASSAYS SUGGEST THAT ACETALDEHYDE FORMS ADDUCTS WITH HISTONE PROTEINS IN HUMAN BRONCHIAL EPITHELIAL BEAS-2B CELLS. THE LEVEL OF ACETYLATION FOR N-TERMINAL TAILS OF CYTOSOLIC HISTONES H3 AND H4, AN IMPORTANT MODIFICATION FOR HISTONE NUCLEAR IMPORT AND CHROMATIN ASSEMBLY, IS SIGNIFICANTLY DOWNREGULATED FOLLOWING ACETALDEHYDE EXPOSURE IN BEAS-2B CELLS, POSSIBLY DUE TO THE FORMATION OF HISTONE ADDUCTS AND/OR THE DECREASE IN THE EXPRESSION OF HISTONE ACETYLTRANSFERASES. NOTABLY, THE LEVEL OF NUCLEOSOMAL HISTONES IN THE CHROMATIN FRACTION AND AT MOST OF THE GENOMIC LOCI WE TESTED ARE LOW IN ACETALDEHYDE-TREATED CELLS AS COMPARED WITH THE CONTROL CELLS, WHICH IS SUGGESTIVE OF INHIBITION OF CHROMATIN ASSEMBLY. MOREOVER, ACETALDEHYDE EXPOSURE PERTURBS CHROMATIN STRUCTURE AS EVIDENCED BY THE INCREASE IN GENERAL CHROMATIN ACCESSIBILITY AND THE DECREASE IN NUCLEOSOME OCCUPANCY AT GENOMIC LOCI FOLLOWING ACETALDEHYDE TREATMENT. OUR RESULTS INDICATE THAT REGULATION OF HISTONE MODIFICATIONS AND CHROMATIN ACCESSIBILITY MAY PLAY IMPORTANT ROLES IN ACETALDEHYDE-INDUCED PATHOGENESIS. ENVIRON. MOL. MUTAGEN. 59:375-385, 2018. (C) 2018 WILEY PERIODICALS, INC.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
8  3619  36 IN VIVO ACUTE ON CHRONIC ETHANOL EFFECTS IN LIVER: A MOUSE MODEL EXHIBITING EXACERBATED INJURY, ALTERED METABOLIC AND EPIGENETIC RESPONSES. CHRONIC ALCOHOLICS WHO ALSO BINGE DRINK (I.E., ACUTE ON CHRONIC) ARE PRONE TO AN EXACERBATED LIVER INJURY BUT ITS MECHANISM IS NOT UNDERSTOOD. WE THEREFORE INVESTIGATED THE IN VIVO EFFECTS OF CHRONIC AND BINGE ETHANOL INGESTION AND COMPARED TO CHRONIC ETHANOL FOLLOWED BY THREE REPEAT BINGE ETHANOL ON THE LIVER OF MALE C57/BL6 MICE FED ETHANOL IN LIQUID DIET (4%) FOR FOUR WEEKS FOLLOWED BY BINGE ETHANOL (INTRAGASTRIC ADMINISTRATION, 3.5 G/KG BODY WEIGHT, THREE DOSES, 12H APART). CHRONIC FOLLOWED BY BINGE ETHANOL EXACERBATED FAT ACCUMULATION, NECROSIS, DECREASE IN HEPATIC SAM AND SAM:SAH RATIO, INCREASE IN ADENOSINE LEVELS, AND ELEVATED CYP2E1 LEVELS. HISTONE H3 LYSINE ACETYLATION (H3ACK9), DUALLY MODIFIED PHOSPHOACETYLATED HISTONE H3 (H3ACK9/PS10), AND PHOSPHORYLATED H2AX INCREASED AFTER BINGE WHEREAS PHOSPHORYLATION OF HISTONE H3 SER 10 (H3S10) AND H3 SER 28 (H3S28) INCREASED AFTER CHRONIC ETHANOL-BINGE. HISTONE H3 LYSINE 4 AND 9 DIMETHYLATION INCREASED WITH A MARKED DIMETHYLATION IN H3K9 IN CHRONIC ETHANOL BINGE GROUP. TRIMETHYLATED HISTONE H3 LEVELS DID NOT CHANGE. NUCLEAR LEVELS OF HISTONE ACETYL TRANSFERASE GCN5 AND HISTONE DEACETYLASE HDAC3 WERE ELEVATED WHEREAS PHOSPHO-CREB DECREASED IN A DISTINCTIVE MANNER. TAKEN TOGETHER, ACUTE ON CHRONIC ETHANOL INGESTION CAUSED AMPLIFICATION OF LIVER INJURY AND ELICITED CHARACTERISTIC PROFILES OF HISTONE MODIFICATIONS, METABOLIC ALTERATIONS, AND CHANGES IN NUCLEAR PROTEIN LEVELS. THESE FINDINGS DEMONSTRATE THAT CHRONIC ETHANOL EXPOSURE RENDERS LIVER MORE SUSCEPTIBLE TO REPEAT ACUTE/BINGE ETHANOL INDUCED ACCELERATION OF ALCOHOLIC LIVER DISEASE.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
9  1837  38 EFFECTS OF PALMITATE ON GENOME-WIDE MRNA EXPRESSION AND DNA METHYLATION PATTERNS IN HUMAN PANCREATIC ISLETS. BACKGROUND: CIRCULATING FREE FATTY ACIDS ARE OFTEN ELEVATED IN PATIENTS WITH TYPE 2 DIABETES (T2D) AND OBESE INDIVIDUALS. CHRONIC EXPOSURE TO HIGH LEVELS OF SATURATED FATTY ACIDS HAS DETRIMENTAL EFFECTS ON ISLET FUNCTION AND INSULIN SECRETION. ALTERED GENE EXPRESSION AND EPIGENETICS MAY CONTRIBUTE TO T2D AND OBESITY. HOWEVER, THERE IS LIMITED INFORMATION ON WHETHER FATTY ACIDS ALTER THE GENOME-WIDE TRANSCRIPTOME PROFILE IN CONJUNCTION WITH DNA METHYLATION PATTERNS IN HUMAN PANCREATIC ISLETS. TO DISSECT THE MOLECULAR MECHANISMS LINKING LIPOTOXICITY TO IMPAIRED INSULIN SECRETION, WE INVESTIGATED THE EFFECTS OF A 48 H PALMITATE TREATMENT IN VITRO ON GENOME-WIDE MRNA EXPRESSION AND DNA METHYLATION PATTERNS IN HUMAN PANCREATIC ISLETS. METHODS: GENOME-WIDE MRNA EXPRESSION WAS ANALYZED USING AFFYMETRIX GENECHIP((R)) HUMAN GENE 1.0 ST WHOLE TRANSCRIPT-BASED ARRAY (N = 13) AND GENOME-WIDE DNA METHYLATION WAS ANALYZED USING INFINIUM HUMANMETHYLATION450K BEADCHIP (N = 13) IN HUMAN PANCREATIC ISLETS EXPOSED TO PALMITATE OR CONTROL MEDIA FOR 48 H. A NON-PARAMETRIC PAIRED WILCOXON STATISTICAL TEST WAS USED TO ANALYZE MRNA EXPRESSION. APOPTOSIS WAS MEASURED USING APO-ONE((R)) HOMOGENEOUS CASPASE-3/7 ASSAY (N = 4). RESULTS: WHILE GLUCOSE-STIMULATED INSULIN SECRETION WAS DECREASED, THERE WAS NO SIGNIFICANT EFFECT ON APOPTOSIS IN HUMAN ISLETS EXPOSED TO PALMITATE. WE IDENTIFIED 1,860 DIFFERENTIALLY EXPRESSED GENES IN PALMITATE-TREATED HUMAN ISLETS. THESE INCLUDE CANDIDATE GENES FOR T2D, SUCH AS TCF7L2, GLIS3, HNF1B AND SLC30A8. ADDITIONALLY, GENES IN GLYCOLYSIS/GLUCONEOGENESIS, PYRUVATE METABOLISM, FATTY ACID METABOLISM, GLUTATHIONE METABOLISM AND ONE CARBON POOL BY FOLATE WERE DIFFERENTIALLY EXPRESSED IN PALMITATE-TREATED HUMAN ISLETS. PALMITATE TREATMENT ALTERED THE GLOBAL DNA METHYLATION LEVEL AND DNA METHYLATION LEVELS OF CPG ISLAND SHELVES AND SHORES, 5'UTR, 3'UTR AND GENE BODY REGIONS IN HUMAN ISLETS. MOREOVER, 290 GENES WITH DIFFERENTIAL EXPRESSION HAD A CORRESPONDING CHANGE IN DNA METHYLATION, FOR EXAMPLE, TCF7L2 AND GLIS3. IMPORTANTLY, OUT OF THE GENES DIFFERENTIALLY EXPRESSED DUE TO PALMITATE TREATMENT IN HUMAN ISLETS, 67 WERE ALSO ASSOCIATED WITH BMI AND 37 WERE DIFFERENTIALLY EXPRESSED IN ISLETS FROM T2D PATIENTS. CONCLUSION: OUR STUDY DEMONSTRATES THAT PALMITATE TREATMENT OF HUMAN PANCREATIC ISLETS GIVES RISE TO EPIGENETIC MODIFICATIONS THAT TOGETHER WITH ALTERED GENE EXPRESSION MAY CONTRIBUTE TO IMPAIRED INSULIN SECRETION AND T2D.	2014	

10  2120  32 EPIGENETIC HISTONE MODIFICATIONS IN A CLINICALLY RELEVANT RAT MODEL OF CHRONIC ETHANOL-BINGE-MEDIATED LIVER INJURY. PURPOSE: ETHANOL BINGE AUGMENTS LIVER INJURY AFTER CHRONIC ETHANOL CONSUMPTION IN HUMANS, BUT THE MECHANISM BEHIND THE ENHANCED LIVER INJURY BY ETHANOL BINGE IS NOT KNOWN. IN THIS STUDY WE USED A CLINICALLY RELEVANT RAT MODEL IN WHICH LIVER INJURY IS AMPLIFIED BY BINGE AFTER CHRONIC ETHANOL TREATMENT AND INVESTIGATED THE IMPORTANCE OF HISTONE MODIFICATIONS. METHODS: EIGHT-WEEK-OLD SPRAGUE-DAWLEY RATS WERE FED ETHANOL IN A LIQUID DIET FOR 4 WEEKS. CONTROL RATS WERE FED AN ISOCALORIC LIQUID DIET. THIS WAS FOLLOWED BY THREE BINGE ADMINISTRATIONS OF ETHANOL (INTRAGASTRIC 5 G/KG BODY WEIGHT, 12 H APART). IN THE CONTROL, ETHANOL WAS REPLACED BY WATER. FOUR HOURS AFTER THE LAST BINGE ADMINISTRATION, LIVER SAMPLES WERE ANALYZED FOR HISTONE MODIFICATIONS AND PARAMETERS OF LIVER INJURY. RESULTS: CHRONIC ETHANOL ADMINISTRATION ALONE CAUSED AN INCREASE IN HISTONE H3 SER10 AND SER28 (H3S10 OR S28) PHOSPHORYLATION, AND BINGE ETHANOL REDUCED THEIR LEVELS. LEVELS OF DUALLY MODIFIED PHOSPHOACETYLATED HISTONE H3 (H3ACK9/PS10) INCREASED AFTER ACUTE BINGE ETHANOL AND REMAINED SAME AFTER CHRONIC ETHANOL BINGE. IN CONTRAST, HISTONE H3 LYSINE-9 ACETYLATION (H3ACK9) WAS NOT INCREASED AFTER CHRONIC ETHANOL BUT INCREASED SIGNIFICANTLY AFTER ACUTE BINGE AND CHRONIC ETHANOL BINGE. INCREASE IN HISTONE ACETYLATION WAS ACCOMPANIED BY INCREASED PHOSPHO-ERK1/2 IN THE NUCLEAR EXTRACTS. INCREASED ACETYLATION AFTER CHRONIC ETHANOL BINGE WAS ALSO ACCOMPANIED BY INCREASED PROTEIN LEVELS OF GCN5 HISTONE ACETYL TRANSFERASE AND A MODEST INCREASE IN HDAC3 IN THE NUCLEUS. HISTONE LYSINE-9 DIMETHYLATION WAS SIGNIFICANTLY INCREASED AFTER CHRONIC ETHANOL BINGE. CHRONIC ETHANOL BINGE ALSO RESULTED IN A DECREASE IN THE SAM:SAH RATIO WITH A RELATIVE DECREASE OF SAM LEVELS AND A CORRESPONDING INCREASE IN SAH LEVELS. CONCLUSIONS: ETHANOL BINGE AFTER CHRONIC ETHANOL ALTERED THE PROFILE OF SITE-SPECIFIC HISTONE MODIFICATIONS AND MAY UNDERLIE THE MECHANISM OF AUGMENTED LIVER INJURY BY CHRONIC-ETHANOL-BINGE-TREATED RATS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
11  2156  43 EPIGENETIC MECHANISMS ARE INVOLVED IN THE REGULATION OF ETHANOL CONSUMPTION IN MICE. BACKGROUND: REPEATED ALCOHOL EXPOSURE IS KNOWN TO INCREASE SUBSEQUENT ETHANOL CONSUMPTION IN MICE. HOWEVER, THE UNDERLYING MECHANISMS HAVE NOT BEEN FULLY ELUCIDATED. ONE POSTULATED MECHANISM INVOLVES EPIGENETIC MODIFICATIONS, INCLUDING HISTONE MODIFICATIONS AND DNA METHYLATION OF RELEVANT GENES SUCH AS NR2B OR BDNF. METHODS: TO INVESTIGATE THE ROLE OF EPIGENETIC MECHANISMS IN THE DEVELOPMENT OF ALCOHOL DRINKING BEHAVIOR, AN ESTABLISHED CHRONIC INTERMITTENT ETHANOL EXPOSURE REINFORCED ETHANOL DRINKING MOUSE MODEL WITH VAPOR INHALATION OVER TWO 9-DAY TREATMENT REGIMENS WAS USED. THE DNA METHYLTRANSFERASE INHIBITOR, 5-AZACYTIDINE OR THE HISTONE DEACETYLASE INHIBITOR, TRICHOSTATIN A WAS ADMINISTERED (INTRAPERITONEALLY) TO C57BL/6 MICE 30 MIN BEFORE DAILY EXPOSURE TO CHRONIC INTERMITTENT ETHANOL. CHANGES IN ETHANOL CONSUMPTION WERE MEASURED USING THE 2-BOTTLE CHOICE TEST. RESULTS: THE RESULTS INDICATED THAT SYSTEMIC ADMINISTRATION OF TRICHOSTATIN A (2.5 MICROG/G) FACILITATED CHRONIC INTERMITTENT ETHANOL-INDUCED ETHANOL DRINKING, BUT SYSTEMIC ADMINISTRATION OF 5-AZACYTIDINE (2 MICROG/G) DID NOT CAUSE THE SAME EFFECT. HOWEVER, WHEN 5-AZACYTIDINE WAS ADMINISTERED BY INTRACEREBROVENTRICULAR INJECTION, IT FACILITATED CHRONIC INTERMITTENT ETHANOL-INDUCED ETHANOL DRINKING. FURTHERMORE, THE INCREASED DRINKING CAUSED BY CHRONIC INTERMITTENT ETHANOL WAS PREVENTED BY INJECTION OF A METHYL DONOR, S-ADENOSYL-L-METHIONINE. TO PROVIDE EVIDENCE THAT CHRONIC INTERMITTENT ETHANOL- OR TRICHOSTATIN A-INDUCED DNA DEMETHYLATION AND HISTONE MODIFICATIONS OF THE NR2B PROMOTER MAY UNDERLIE THE ALTERED ETHANOL CONSUMPTION, WE EXAMINED EPIGENETIC MODIFICATIONS AND NR2B EXPRESSION IN THE PREFRONTAL CORTEX OF THESE MICE. CHRONIC INTERMITTENT ETHANOL OR TRICHOSTATIN A DECREASED DNA METHYLATION AND INCREASED HISTONE ACETYLATION IN THE NR2B GENE PROMOTER, AS WELL AS MRNA LEVELS OF NR2B IN THESE MICE. CONCLUSIONS: TAKEN TOGETHER, THESE RESULTS INDICATE THAT EPIGENETIC MODIFICATIONS ARE INVOLVED IN REGULATING ETHANOL DRINKING BEHAVIOR, PARTIALLY THROUGH ALTERING NR2B EXPRESSION.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                 
12  4943  33 PATERNAL PRECONCEPTION ALCOHOL EXPOSURE IMPARTS INTERGENERATIONAL ALCOHOL-RELATED BEHAVIORS TO MALE OFFSPRING ON A PURE C57BL/6J BACKGROUND. WHILE ALCOHOL USE DISORDER (AUD) IS A HIGHLY HERITABLE CONDITION, THE BASIS OF AUD IN FAMILIES WITH A HISTORY OF ALCOHOLISM IS DIFFICULT TO EXPLAIN BY GENETIC VARIATION ALONE. EMERGING EVIDENCE SUGGESTS THAT PARENTAL EXPERIENCE PRIOR TO CONCEPTION CAN AFFECT INHERITANCE OF COMPLEX BEHAVIORS IN OFFSPRING VIA NON-GENOMIC (EPIGENETIC) MECHANISMS. FOR INSTANCE, MALE C57BL/6J (B6) MICE EXPOSED TO CHRONIC INTERMITTENT VAPOR ETHANOL (CIE) PRIOR TO MATING WITH STRAIN 129S1/SVIMJ ETHANOL-NAIVE FEMALES PRODUCE MALE OFFSPRING WITH REDUCED ETHANOL-DRINKING PREFERENCE, INCREASED ETHANOL SENSITIVITY, AND INCREASED BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) EXPRESSION IN THE VENTRAL TEGMENTAL AREA (VTA). IN THE PRESENT STUDY, WE TESTED THE HYPOTHESIS THAT THESE INTERGENERATIONAL EFFECTS OF PATERNAL CIE ARE REPRODUCIBLE IN MALE OFFSPRING ON AN INBRED B6 BACKGROUND. TO THIS END, B6 MALES WERE EXPOSED TO 6 WEEKS OF CIE (OR ROOM AIR AS A CONTROL) BEFORE MATING WITH ETHANOL-NAIVE B6 FEMALES TO PRODUCE ETHANOL (E)-SIRED AND CONTROL (C)-SIRED MALE AND FEMALE OFFSPRING. WE OBSERVED A SEX-SPECIFIC EFFECT, AS E-SIRED MALES EXHIBITED DECREASED TWO-BOTTLE FREE-CHOICE ETHANOL-DRINKING PREFERENCE, INCREASED SENSITIVITY TO THE ANXIOLYTIC EFFECTS OF ETHANOL, AND INCREASED VTA BDNF EXPRESSION; NO DIFFERENCES WERE OBSERVED IN FEMALE OFFSPRING. THESE FINDINGS CONFIRM AND EXTEND OUR PREVIOUS RESULTS BY DEMONSTRATING THAT THE EFFECTS OF PATERNAL PRECONCEPTION ETHANOL ARE REPRODUCIBLE USING GENETICALLY IDENTICAL, INBRED B6 ANIMALS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
13  6177  31 THE HISTONE METHYLTRANSFERASE G9A MEDIATES STRESS-REGULATED ALCOHOL DRINKING. THE EPIGENETIC ENZYME G9A IS A HISTONE METHYLTRANSFERASE THAT DIMETHYLATES LYSINE 9 ON HISTONE H3 (H3K9ME2), AND IN THE ADULT NUCLEUS ACCUMBENS (NAC), G9A REGULATES MULTIPLE BEHAVIORS ASSOCIATED WITH SUBSTANCE USE DISORDER. WE SHOW HERE THAT CHRONIC INTERMITTENT ETHANOL (CIE) EXPOSURE IN MALE MICE REDUCED BOTH G9A AND H3K9ME2 LEVELS IN THE ADULT NAC, BUT NOT DORSAL STRIATUM. VIRAL-MEDIATED REDUCTION OF G9A IN THE NAC HAD NO EFFECTS ON BASELINE VOLITIONAL ETHANOL DRINKING OR ESCALATED ALCOHOL DRINKING PRODUCED BY CIE EXPOSURE; HOWEVER, NAC G9A WAS REQUIRED FOR STRESS-REGULATED CHANGES IN ETHANOL DRINKING, INCLUDING POTENTIATED ALCOHOL DRINKING PRODUCED BY ACTIVATION OF THE KAPPA-OPIOID RECEPTOR. IN ADDITION, WE OBSERVED THAT CHRONIC SYSTEMIC ADMINISTRATION OF A G9A INHIBITOR, UNC0642, ALSO BLOCKED STRESS-POTENTIATED ALCOHOL DRINKING. TOGETHER, OUR FINDINGS SUGGEST THAT CHRONIC ALCOHOL USE, SIMILAR TO OTHER ABUSED SUBSTANCES, PRODUCES A NAC-SELECTIVE REDUCTION IN G9A LEVELS THAT SERVES TO LIMIT STRESS-REGULATED ALCOHOL DRINKING. MOREOVER, OUR FINDINGS SUGGEST THAT PHARMACOLOGICAL INHIBITION OF G9A MIGHT PROVIDE A NOVEL THERAPEUTIC APPROACH TO TREAT STRESS-INDUCED ALCOHOL DRINKING, WHICH IS A MAJOR TRIGGER OF RELAPSE IN INDIVIDUALS SUFFERING FROM AUD.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
14   894  32 CHRONIC ETHANOL FEEDING ALTERS HEPATOCYTE MEMORY WHICH IS NOT ALTERED BY ACUTE FEEDING. BACKGROUND: GENE EXPRESSION CHANGES IN THE LIVER AFTER ACUTE BINGE DRINKING MAY DIFFER FROM THE CHANGES SEEN IN CHRONIC ETHANOL FEEDING IN THE RAT. THE CHANGES IN GENE EXPRESSION AFTER CHRONIC ETHANOL FEEDING MAY SENSITIZE THE LIVER TO ALCOHOL-INDUCED LIVER DAMAGE, WHICH IS NOT SEEN AFTER ACUTE BINGE DRINKING. METHODS: TO TEST THIS HYPOTHESIS, GENE MICROARRAY ANALYSIS WAS PERFORMED ON THE LIVERS OF RATS (N = 3) FED AN ACUTE BINGE DOSE OF ETHANOL (6 G/KG BODY WT) AND KILLED AT 3 AND 12 HOURS AFTER ETHANOL BY GAVAGE. THE GENE MICROARRAYS WERE COMPARED WITH THOSE MADE ON THE LIVER OF RATS FROM A PREVIOUS STUDY, IN WHICH THE RATS WERE FED ETHANOL BY INTRAGASTRIC TUBE FOR 1 MONTH (36% OF CALORIES DERIVED FROM ETHANOL). RESULTS: MICROARRAY ANALYSIS DATA VARIED BETWEEN THE ACUTE AND CHRONIC MODELS IN SEVERAL IMPORTANT RESPECTS. GROWTH FACTORS INCREASED MAINLY IN THE CHRONIC ALCOHOL FED RAT. CHANGES IN ENZYMES INVOLVED IN OXIDATIVE STRESS WERE NOTED ONLY WITH CHRONIC ETHANOL FEEDING. GENE EXPRESSION OF FAT METABOLISM WAS INCREASED ONLY WITH CHRONIC ETHANOL FEEDING. MOST IMPORTANTLY, EPIGENETIC RELATED ENZYMES AND ACETYLATION AND METHYLATION OF HISTONES CHANGED ONLY AFTER CHRONIC ETHANOL FEEDING. CONCLUSIONS: THE RESULTS SUPPORT THE CONCEPT THAT CHRONIC ETHANOL INGESTION INDUCES ALTERED GENE EXPRESSION AS A RESULT OF CHANGES IN EPIGENETIC MECHANISMS, WHERE ACETYLATION AND METHYLATION OF HISTONES WERE ALTERED.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
15  5609  41 S-ADENOSYLMETHIONINE DECREASES THE PEAK BLOOD ALCOHOL LEVELS 3 H AFTER AN ACUTE BOLUS OF ETHANOL BY INDUCING ALCOHOL METABOLIZING ENZYMES IN THE LIVER. INTRODUCTION: AN ALCOHOL BOLUS CAUSES THE BLOOD ALCOHOL LEVEL (BAL) TO PEAK AT 1-2 H POST INGESTION. THE ETHANOL ELIMINATION RATE IS REGULATED BY ALCOHOL METABOLIZING ENZYMES, PRIMARILY ALCOHOL DEHYDROGENASE (ADH1), ACETALDEHYDE DEHYDROGENASE (ALDH), AND CYTOCHROME P450 (CYP2E1). RECENTLY, S-ADENOSYLMETHIONINE (SAME) WAS FOUND TO REDUCE ACUTE BALS 3 H AFTER AN ALCOHOL BOLUS. THE QUESTION, THEN, WAS: WHAT IS THE MECHANISM INVOLVED IN THIS REDUCTION OF BAL BY FEEDING SAME? TO ANSWER THIS QUESTION, WE INVESTIGATED THE CHANGES IN ETHANOL METABOLIZING ENZYMES AND THE EPIGENETIC CHANGES THAT REGULATE THE EXPRESSION OF THESE ENZYMES DURING ACUTE BINGE DRINKING AND CHRONIC DRINKING. METHODS: RATS WERE FED A BOLUS OF ETHANOL WITH OR WITHOUT SAME, AND WERE SACRIFICED AT 3 H OR 12 H AFTER THE BOLUS. RESULTS: RT-PCR AND WESTERN BLOT ANALYSES SHOWED THAT SAME SIGNIFICANTLY INDUCED ADH1 LEVELS IN THE 3 H LIVER SAMPLES. HOWEVER, SAME DID NOT AFFECT THE CHANGES IN ADH1 PROTEIN LEVELS 12 H POST BOLUS. SINCE SAME IS A METHYL DONOR, IT WAS POSTULATED THAT THE ADH1 GENE EXPRESSION UP REGULATION AT 3 H WAS DUE TO A HISTONE MODIFICATION INDUCED BY METHYLATION FROM METHYL TRANSFERASES. DIMETHYLATED HISTONE 3 LYSINE 4 (H3K4ME2), A MODIFICATION RESPONSIBLE FOR GENE EXPRESSION ACTIVATION, WAS FOUND TO BE SIGNIFICANTLY INCREASED BY SAME AT 3 H POST BOLUS. CONCLUSION: THESE RESULTS CORRELATED WITH THE LOW BAL FOUND AT 3 H POST BOLUS, AND SUPPORT THE CONCEPT THAT SAME INCREASED THE GENE EXPRESSION TO INCREASE THE ELIMINATION RATE OF ETHANOL IN BINGE DRINKING BY INCREASING H3K4ME2.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
16  4931  36 PATERNAL ALCOHOL EXPOSURE REDUCES ALCOHOL DRINKING AND INCREASES BEHAVIORAL SENSITIVITY TO ALCOHOL SELECTIVELY IN MALE OFFSPRING. ALCOHOL USE DISORDER (AUD) IS HERITABLE, BUT THE GENETIC BASIS FOR THIS DISEASE REMAINS POORLY UNDERSTOOD. ALTHOUGH NUMEROUS GENE VARIANTS HAVE BEEN ASSOCIATED WITH AUD, THESE VARIANTS ACCOUNT FOR ONLY A SMALL FRACTION OF THE TOTAL RISK. THE IDEA OF INHERITANCE OF ACQUIRED CHARACTERISTICS, I.E. "EPIGENETIC INHERITANCE," IS RE-EMERGING AS A PROVEN ADJUNCT TO TRADITIONAL MODES OF GENETIC INHERITANCE. WE HYPOTHESIZED THAT ALCOHOL DRINKING AND NEUROBIOLOGICAL SENSITIVITY TO ALCOHOL ARE INFLUENCED BY ANCESTRAL ALCOHOL EXPOSURE. TO TEST THIS HYPOTHESIS, WE EXPOSED MALE MICE TO CHRONIC VAPOR ETHANOL OR CONTROL CONDITIONS, MATED THEM TO ETHANOL-NAIVE FEMALES, AND TESTED ADULT OFFSPRING FOR ETHANOL DRINKING, ETHANOL-INDUCED BEHAVIORS, GENE EXPRESSION, AND DNA METHYLATION. WE FOUND THAT ETHANOL-SIRED MALE OFFSPRING HAD REDUCED ETHANOL PREFERENCE AND CONSUMPTION, ENHANCED SENSITIVITY TO THE ANXIOLYTIC AND MOTOR-ENHANCING EFFECTS OF ETHANOL, AND INCREASED BDNF EXPRESSION IN THE VENTRAL TEGMENTAL AREA (VTA) COMPARED TO CONTROL-SIRED MALE OFFSPRING. THERE WERE NO DIFFERENCES AMONG ETHANOL- AND CONTROL-SIRED FEMALE OFFSPRING ON THESE ASSAYS. ETHANOL EXPOSURE ALSO DECREASED DNA METHYLATION AT THE BDNFAEPROMOTER OF SIRE'S GERM CELLS AND HYPOMETHYLATION WAS MAINTAINED IN THE VTA OF BOTH MALE AND FEMALE ETHANOL-SIRED OFFSPRING. OUR FINDINGS SHOW THAT PATERNAL ALCOHOL EXPOSURE IS A PREVIOUSLY UNRECOGNIZED REGULATOR OF ALCOHOL DRINKING AND BEHAVIORAL SENSITIVITY TO ALCOHOL IN MALE, BUT NOT FEMALE, OFFSPRING. PATERNAL ALCOHOL EXPOSURE ALSO INDUCES EPIGENETIC ALTERATIONS (DNA HYPOMETHYLATION) AND GENE EXPRESSION CHANGES THAT PERSIST IN THE VTA OF OFFSPRING. THESE RESULTS PROVIDE NEW INSIGHT INTO THE INHERITANCE AND DEVELOPMENT OF ALCOHOL DRINKING BEHAVIORS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
17  5445  40 REPEATED VAPOR ETHANOL EXPOSURE INDUCES TRANSIENT HISTONE MODIFICATIONS IN THE BRAIN THAT ARE MODIFIED BY GENOTYPE AND BRAIN REGION. BACKGROUND: EMERGING RESEARCH IMPLICATES ETHANOL (ETOH)-INDUCED EPIGENETIC MODIFICATIONS IN REGULATING GENE EXPRESSION AND ETOH CONSUMPTION. HOWEVER, CONSENSUS ON SPECIFIC EPIGENETIC MODIFICATIONS INDUCED BY ETOH HAS NOT YET EMERGED, MAKING IT CHALLENGING TO IDENTIFY MECHANISMS AND DEVELOP TARGETED TREATMENTS. WE HYPOTHESIZED THAT CHRONIC INTERMITTENT ETOH (CIE) INDUCES PERSISTENT CHANGES IN HISTONE MODIFICATIONS ACROSS THE CEREBRAL CORTEX (CCX), NUCLEUS ACCUMBENS (NAC), AND PREFRONTAL CORTEX (PFC), AND THAT THESE HISTONE MODIFICATIONS ARE ALTERED IN A KNOCK-IN MOUSE STRAIN WITH ALTERED SENSITIVITY TO ETOH. METHODS: C57BL/6J (B6) MICE AND ALPHA1SHLA KNOCKIN MICE ON A B6 BACKGROUND WERE EXPOSED TO 16 H OF VAPOR ETOH OR ROOM AIR FOLLOWED BY 8 H OF ROOM AIR FOR 4 CONSECUTIVE DAYS AND SACRIFICED AT MULTIPLE TIME POINTS UP TO 72 H FOLLOWING EXPOSURE. HISTONE MODIFICATIONS WERE ASSESSED USING WESTERN BLOT AND DOT BLOT. RT-QPCR WAS USED TO STUDY EXPRESSION OF CHROMATIN MODIFYING ENZYMES IN NAC AND PFC. RESULTS: IN NAC, CIE SIGNIFICANTLY INCREASED ACETYLATION OF HISTONE SUBUNIT H3 AT LYSINE 9 (H3K9AC) BUT NOT LYSINE 14 (H3K14AC) OR LYSINE 27 (H3K27AC). IN PFC, CIE SIGNIFICANTLY INCREASED H3K9AC BUT NOT H3K14 OR H3K27AC. THERE WERE NO SIGNIFICANT CHANGES AT 8 OR 72 H AFTER ETOH EXPOSURE IN EITHER NAC OR PFC. CIE WAS ALSO ASSOCIATED WITH INCREASED EXPRESSION OF KAT2B, KAT5, AND TET1 IN NAC BUT NOT PFC. IN CCX, CIE HAD A SIGNIFICANT EFFECT ON LEVELS OF H3K18AC; THERE WAS ALSO A SIGNIFICANT EFFECT OF THE ALPHA1SHLA MUTATION ON LEVELS OF H3K27ME3, H3K14AC, AND H3K18AC AS WELL AS A TREND FOR H3S10PK14AC. CONCLUSIONS: THE ETOH-INDUCED HISTONE MODIFICATIONS OBSERVED WERE TRANSIENT AND VARIED SIGNIFICANTLY BETWEEN BRAIN REGIONS. A GENETIC MUTATION THAT ALTERED SENSITIVITY TO ETOH WAS ASSOCIATED WITH ALTERED INDUCTION OF HISTONE MODIFICATIONS DURING CIE. THESE RESULTS HAVE IMPLICATIONS FOR STUDYING ETOH-INDUCED HISTONE MODIFICATIONS AND ETOH SENSITIVITY.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
18  3841  33 IRON SUPPLEMENTATION REVERSES THE REDUCTION OF HYDROXYMETHYLCYTOSINE IN HEPATIC DNA ASSOCIATED WITH CHRONIC ALCOHOL CONSUMPTION IN RATS. BACKGROUND: ALCOHOL IS KNOWN TO AFFECT TWO EPIGENETIC PHENOMENA, DNA METHYLATION AND DNA HYDROXYMETHYLATION, AND IRON IS A COFACTOR OF TEN-ELEVEN TRANSLOCATION (TET) ENZYMES THAT CATALYZE THE CONVERSION FROM METHYLCYTOSINE TO HYDROXYMETHYLCYTOSINE. IN THE PRESENT STUDY WE AIMED TO DETERMINE THE EFFECTS OF ALCOHOL ON DNA HYDROXYMETHYLATION AND FURTHER EFFECTS OF IRON ON ALCOHOL ASSOCIATED EPIGENETIC CHANGES. METHODS: TWENTY-FOUR MALE SPRAGUE-DAWLEY RATS WERE FED EITHER LIEBER-DECARLI ALCOHOL DIET (36% CALORIES FROM ETHANOL) OR LIEBER-DECARLI CONTROL DIET ALONG WITH OR WITHOUT IRON SUPPLEMENTATION (0.6% CARBONYL IRON) FOR 8 WEEKS. HEPATIC NON-HEME IRON CONCENTRATIONS WERE MEASURED BY COLORIMETRIC ASSAYS. PROTEIN LEVELS OF HEPATIC FERRITIN AND TRANSFERRIN RECEPTOR WERE DETERMINED BY WESTERN BLOTTING. METHYLCYTOSINE, HYDROXYMETHYLCYTOSINE AND UNMODIFIED CYTOSINE IN DNA WERE SIMULTANEOUSLY MEASURED BY LIQUID CHROMATOGRAPHY/MASS SPECTROMETRY METHOD. RESULTS: IRON SUPPLEMENTATION SIGNIFICANTLY INCREASED HEPATIC NON-HEME IRON CONTENTS (P < 0.05) BUT ALCOHOL ALONE DID NOT. HOWEVER, BOTH ALCOHOL AND IRON SIGNIFICANTLY INCREASED HEPATIC FERRITIN LEVELS AND DECREASED HEPATIC TRANSFERRIN RECEPTOR LEVELS (P < 0.05). ALCOHOL REDUCED HEPATIC DNA HYDROXYMETHYLATION (0.21% +/- 0.04% VS. 0.33% +/- 0.04%, P = 0.01) COMPARED TO CONTROL, WHILE IRON SUPPLEMENTATION TO ALCOHOL DIET DID NOT CHANGE DNA HYDROXYMETHYLATION. THERE WAS NO SIGNIFICANT DIFFERENCE IN METHYLCYTOSINE LEVELS, WHILE UNMODIFIED CYTOSINE LEVELS WERE SIGNIFICANTLY INCREASED IN ALCOHOL-FED GROUPS COMPARED TO CONTROL (95.61% +/- 0.08% VS. 95.26% +/- 0.12%, P = 0.03), SUGGESTING THAT ALCOHOL FURTHER INCREASES THE CONVERSION FROM HYDROXYMETHYLCYTOSINE TO UNMODIFIED CYTOSINE. CONCLUSIONS: CHRONIC ALCOHOL CONSUMPTION ALTERS GLOBAL DNA HYDROXYMETHYLATION IN THE LIVER BUT IRON SUPPLEMENTATION REVERSES THE EPIGENETIC EFFECT OF ALCOHOL.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
19  6639  36 UNRAVELING THE EPIGENOMIC AND TRANSCRIPTOMIC INTERPLAY DURING ALCOHOL-INDUCED ANXIOLYSIS. POSITIVE EFFECTS OF ALCOHOL DRINKING SUCH AS ANXIOLYSIS AND EUPHORIA APPEAR TO BE A CRUCIAL FACTOR IN THE INITIATION AND MAINTENANCE OF ALCOHOL USE DISORDER (AUD). HOWEVER, THE MECHANISMS THAT LEAD FROM CHROMATIN REORGANIZATION TO TRANSCRIPTOMIC CHANGES AFTER ACUTE ETHANOL EXPOSURE REMAIN UNKNOWN. HERE, WE USED ASSAY FOR TRANSPOSASE-ACCESSIBLE CHROMATIN FOLLOWED BY HIGH THROUGHPUT SEQUENCING (ATAC-SEQ) AND RNA-SEQ TO INVESTIGATE EPIGENOMIC AND TRANSCRIPTOMIC CHANGES THAT UNDERLIE ANXIOLYTIC EFFECTS OF ACUTE ETHANOL USING AN ANIMAL MODEL. ANALYSIS OF ATAC-SEQ DATA REVEALED AN OVERALL OPEN OR PERMISSIVE CHROMATIN STATE THAT WAS ASSOCIATED WITH TRANSCRIPTOMIC CHANGES IN THE AMYGDALA AFTER ACUTE ETHANOL EXPOSURE. WE IDENTIFIED A CANDIDATE GENE, HIF3A (HYPOXIA-INDUCIBLE FACTOR 3, ALPHA SUBUNIT), THAT HAD 'OPEN' CHROMATIN REGIONS (ATAC-SEQ PEAKS), ASSOCIATED WITH SIGNIFICANTLY INCREASED ACTIVE EPIGENETIC HISTONE ACETYLATION MARKS AND DECREASED DNA METHYLATION AT THESE REGIONS. THE MRNA LEVELS OF HIF3A WERE INCREASED BY ACUTE ETHANOL EXPOSURE, BUT DECREASED IN THE AMYGDALA DURING WITHDRAWAL AFTER CHRONIC ETHANOL EXPOSURE. KNOCKDOWN OF HIF3A EXPRESSION IN THE CENTRAL NUCLEUS OF AMYGDALA ATTENUATED ACUTE ETHANOL-INDUCED INCREASES IN HIF3A MRNA LEVELS AND BLOCKED ANXIOLYSIS IN RATS. THESE DATA INDICATE THAT CHROMATIN ACCESSIBILITY AND TRANSCRIPTOMIC SIGNATURES IN THE AMYGDALA AFTER ACUTE ETHANOL EXPOSURE UNDERLIE ANXIOLYSIS AND POSSIBLY PRIME THE CHROMATIN FOR THE DEVELOPMENT OF AUD.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
20   312  42 ALCOHOL FEEDING IN MICE PROMOTES COLONIC HYPERPERMEABILITY AND CHANGES IN COLONIC ORGANOID STEM CELL FATE. BACKGROUND: ALCOHOL INCREASES INTESTINAL PERMEABILITY TO PROINFLAMMATORY MICROBIAL PRODUCTS THAT PROMOTE LIVER DISEASE, EVEN AFTER A PERIOD OF SOBRIETY. WE SOUGHT TO TEST THE HYPOTHESIS THAT ALCOHOL AFFECTS INTESTINAL STEM CELLS USING AN IN VIVO MODEL AND EX VIVO ORGANOIDS GENERATED FROM JEJUNUM AND COLON FROM MICE FED CHRONIC ALCOHOL. METHODS: MICE WERE FED A CONTROL OR AN ALCOHOL DIET. INTESTINAL PERMEABILITY, LIVER STEATOSIS-INFLAMMATION, AND STOOL SHORT-CHAIN FATTY ACIDS (SCFAS) WERE MEASURED. JEJUNUM AND COLONIC ORGANOIDS AND TISSUE WERE STAINED FOR STEM CELL, CELL LINEAGE, AND APICAL JUNCTION MARKERS WITH ASSESSMENT OF MRNA BY PCR AND RNA-SEQ. CHIP-PCR ANALYSIS WAS CARRIED OUT FOR NOTCH1 USING AN ANTIBODY SPECIFIC FOR ACETYLATED HISTONE 3. RESULTS: ALCOHOL-FED MICE EXHIBITED COLONIC (BUT NOT SMALL INTESTINAL) HYPERPERMEABILITY, STEATOHEPATITIS, AND DECREASED BUTYRATE/TOTAL SCFA RATIO IN STOOL. STEM CELL, CELL LINEAGE, AND APICAL JUNCTION MARKER STAINING IN TISSUE OR ORGANOIDS FROM JEJUNUM TISSUE WERE NOT IMPACTED BY ALCOHOL. ONLY CHROMOGRANIN A (CHGA) WAS INCREASED IN JEJUNUM ORGANOIDS BY QPCR. HOWEVER, COLONIC TISSUE AND ORGANOID STAINING EXHIBITED AN ALCOHOL-INDUCED SIGNIFICANT DECREASE IN CYTOKERATIN 20+ (KRT20+) ABSORPTIVE LINEAGE ENTEROCYTES, A DECREASE IN OCCLUDIN AND E-CADHERIN APICAL JUNCTION PROTEINS, AN INCREASE IN CHGA, AND AN INCREASE IN THE LGR5 STEM CELL MARKER. QPCR REVEALED AN ALCOHOL-INDUCED DECREASE IN COLONIC ORGANOID AND TISSUE NOTCH1, HES1, AND KRT20 AND INCREASED CHGA, SUPPORTING AN ALTERATION IN STEM CELL FATE DUE TO DECREASED NOTCH1 EXPRESSION. COLONIC TISSUE CHIP-PCR REVEALED ALCOHOL FEEDING SUPPRESSED NOTCH1 MRNA EXPRESSION (VIA DEACETYLATION OF HISTONE H3) AND DECREASED NOTCH1 TISSUE STAINING. CONCLUSIONS: DATA SUPPORT A MODEL FOR ALCOHOL-INDUCED COLONIC HYPERPERMEABILITY VIA EPIGENETIC EFFECTS ON NOTCH1, AND THUS HES1, SUPPRESSION THROUGH A MECHANISM INVOLVING HISTONE H3 DEACETYLATION AT THE NOTCH1 LOCUS. THIS DECREASED ENTEROCYTE AND INCREASED ENTEROENDOCRINE CELL COLONIC STEM CELL FATE AND DECREASED APICAL JUNCTIONAL PROTEINS LEADING TO HYPERPERMEABILITY.	2017