1  2907 148 GENE EXPRESSION MODIFICATIONS IN THE LIVER CAUSED BY BINGE DRINKING AND S-ADENOSYLMETHIONINE FEEDING. THE ROLE OF EPIGENETIC CHANGES. CHRONIC ETHANOL INGESTION, ACHIEVED BY FEEDING ETHANOL AT A CONSTANT RATE USING INTRAGASTRIC TUBE FEEDING, ALTERS THE EXPRESSION OF GENES IN THE LIVER. THIS IS DONE BY EPIGENETIC MECHANISMS, WHICH DEPEND ON THE BLOOD ALCOHOL LEVELS AT THE TIME OF KILLING. HOWEVER, ACUTE BOLUS FEEDING OF ETHANOL CHANGES GENE EXPRESSION WITHOUT LASTING EPIGENETIC CHANGES. THIS OCCURS WITH HISTONE 3 METHYLATION AND ACETYLATION MODIFICATIONS. THE GENE EXPRESSION RESPONSE TO AN ACUTE BOLUS OF ETHANOL MIGHT BE MODIFIED BY FEEDING S-ADENOSYLMETHIONINE (SAME), A METHYL DONOR. IN THE PRESENT STUDY, RATS WERE GIVEN A BOLUS OF ETHANOL (6 G/KG BODY WEIGHT (BW), SAME (1 G/KG BW), ETHANOL + SAME, OR ISOCALORIC GLUCOSE. THE GROUP OF RATS (N = 3) WERE KILLED AT 3 AND 12 H POST BOLUS, AND GENE MICROARRAY ANALYSIS WAS PERFORMED ON THEIR LIVER CELLS. SAME REDUCED THE 3 H BLOOD ETHANOL LEVELS AND INCREASED THE ALT LEVELS AT 3 H. VENN DIAGRAMS SHOWED THAT ALCOHOL CHANGED THE EXPRESSION OF 646 GENES AT 3 H POST BOLUS AND 586 GENES AT 12 H. SAME CHANGED THE EXPRESSION OF 1,012 GENES WHEN FED WITH ETHANOL 3 H POST ETHANOL BOLUS AND 554 GENES AT 12 H POST ETHANOL BOLUS. SAME ALONE CHANGED THE EXPRESSION OF 1,751 GENES AT 3 H AND 1,398 AT 12 H. THERE WERE MORE CHANGES IN GENE EXPRESSION AT 3 H THAN AT 12 H POST ETHANOL WHEN ETHANOL ALONE WAS COMPARED TO THE DEXTROSE CONTROL. THE SAME WAS TRUE WHEN SAME WAS COMPARED TO SAME + ETHANOL. ETHANOL UP REGULATED GENE EXPRESSION IN MOST FUNCTIONAL PATHWAYS AT 3 H. HOWEVER, WHEN SAME WAS FED WITH ETHANOL AT 3 H, MOST PATHWAYS WERE DOWN REGULATED. AT 12 H, HOWEVER, WHEN ETHANOL WAS FED, THE PATHWAYS WERE HALF UP REGULATED AND HALF DOWN REGULATED. THE SAME WAS TRUE WHEN SAME + ETHANOL WAS FED. THE EXPRESSION OF EPIGENETICALLY IMPORTANT GENES, SUCH AS BHMT AND FOXN3, WAS UP REGULATED 3 H POST ALCOHOL BOLUS. AT 3 H, SAME DOWN REGULATED THE EXPRESSION OF GENES, SUCH AS BHMT, MAT2A, JUN, TNFRS9, AHCY 1, TGFBR1 AND 2, AND PCAF. AT 12 H, THE INSULIN SIGNALING PATHWAYS WERE HALF DOWN REGULATED BY ETHANOL, WHICH WAS PARTLY PREVENTED BY SAME. THE MAPK PATHWAY WAS UP REGULATED BY ETHANOL, BUT SAME DID NOT PREVENT THIS. IN CONCLUSION, PROFOUND CHANGES IN GENE EXPRESSION EVOLVED BETWEEN 3 H AND 12 POST ETHANOL BOLUS. SAME DOWN REGULATED THESE CHANGES IN GENE EXPRESSION AT 3 H, AND LESS SO AT 12 H.	2010	

2  1456  38 DISCRIMINATING THE MOLECULAR BASIS OF HEPATOTOXICITY USING THE LARGE-SCALE CHARACTERISTIC MOLECULAR SIGNATURES OF TOXICANTS BY EXPRESSION PROFILING ANALYSIS. PREDICTING THE POTENTIAL HUMAN HEALTH RISK POSED BY CHEMICAL STRESSORS HAS LONG BEEN A MAJOR CHALLENGE FOR TOXICOLOGISTS, AND THE USE OF MICROARRAYS TO MEASURE RESPONSES TO TOXICOLOGICALLY RELEVANT GENES, AND TO IDENTIFY SELECTIVE, SENSITIVE BIOMARKERS OF TOXICITY IS A MAJOR APPLICATION OF PREDICTIVE AND DISCOVERY TOXICOLOGY. TO INVESTIGATE THIS POSSIBILITY, WE INVESTIGATED WHETHER CARCINOGENS (AT DOSES KNOWN TO INDUCE LIVER TUMORS IN CHRONIC EXPOSURE BIOASSAYS) DEREGULATE CHARACTERISTIC SETS OF GENES IN MICE. MALE C3H/HE MICE WERE DOSED WITH TWO HEPATOCARCINOGENS (VINYL CHLORIDE (VC, 50-25 MG/KG), ALDRIN (AD, 0.8-0.4 MG/KG)), OR TWO NON-HEPATOCARCINOGENS (COPPER SULFATE (CS, 150-60 MG/KG), 2,4,5-TRICHLOROPHENOXYACETIC ACID (2,4,5-T, 150-60 MG/KG)). LARGE-SCALE MOLECULAR CHANGES ELICITED BY THESE FOUR HEPATOTOXICANTS IN LIVER TISSUES WERE ANALYZED USING DNA MICROARRAY. THREE DAYS AFTER ADMINISTRATION, NO SIGNIFICANT PHENOTYPIC CHANGES WERE INDUCED BY THESE FOUR DIFFERENT HEPATOTOXICANTS IN TERMS OF HISTOLOGICAL EXAMINATION OR BLOOD BIOCHEMICAL ASSAY. HOWEVER, UNSUPERVISED HIERARCHICAL ANALYSIS OF GENE EXPRESSIONAL CHANGES INDUCED BY HEPATOTOXICANTS RESULTED IN TWO MAJOR GENE SUBCLUSTERS ON DENDROGRAM, I.E., A CARCINOGEN (VN, AD) AND NON-CARCINOGEN GROUP (CS, 2,4,5-T), AND ALSO REVEALED THAT DISTINCT MOLECULAR SIGNATURES EXIST. THESE SIGNATURES WERE FOUNDED ON WELL-DEFINED FUNCTIONAL GENE CATEGORIES AND MAY DIFFERENTIATE GENOTOXIC AND NON-GENOTOXIC CARCINOGENS. FURTHERMORE, VENN DIAGRAM ANALYSIS ALLOWED US TO IDENTIFY CARCINOGEN AND NON-CARCINOGEN-ASSOCIATED MOLECULAR SIGNATURES. USING STATISTICAL METHODS, WE ANALYZED OUTLIER GENES FOR FOUR DIFFERENT CLASSES (GENOTOXIC-, NON-GENOTOXIC-CARCINOGEN, GENOTOXIC-, NON-GENOTOXIC NON-CARCINOGEN) IN TERMS OF THEIR POTENTIAL TO PREDICT DIFFERENT MODES-OF-ACTION. IN CONCLUSION, THE IDENTIFICATION OF LARGE-SCALE MOLECULAR CHANGES IN DIFFERENT HEPATOCARCINOGEN EXPOSURE MODELS REVEALED THAT DIFFERENT TYPES OF HEPATOTOXICANTS ARE ASSOCIATED WITH DIFFERENT EPIGENETIC CHANGES AND MOLECULAR PATHWAYS AND THAT THESE LARGE-SCALE CHARACTERISTIC MOLECULAR CHANGES COULD BE USED AS PREDICTABLE TOXICITY MARKERS.	2008	
                                                                                                                             
3  5609  60 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
4  2120  47 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	
                                                                                                                                                                                                                                                                                                                                                   
5   894  59 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
6   872  42 CHRONIC ALCOHOL EXPOSURE DIFFERENTIALLY ALTERS ONE-CARBON METABOLISM IN RAT LIVER AND BRAIN. BACKGROUND: EPIGENETIC MECHANISMS SUCH AS DNA METHYLATION PLAY AN IMPORTANT ROLE IN REGULATING THE PATHOPHYSIOLOGY OF ALCOHOLISM. CHRONIC ALCOHOL EXPOSURE LEADS TO BEHAVIORAL CHANGES AS WELL AS DECREASED EXPRESSION OF GENES ASSOCIATED WITH SYNAPTIC PLASTICITY. IN THE LIVER, IT HAS BEEN DOCUMENTED THAT CHRONIC ALCOHOL EXPOSURE IMPAIRS METHIONINE SYNTHASE (MS) ACTIVITY LEADING TO A DECREASE IN S-ADENOSYL METHIONINE/S-ADENOSYL HOMOCYSTEINE (SAM/SAH) RATIO WHICH RESULTS IN DNA HYPOMETHYLATION; HOWEVER, IT IS NOT KNOWN WHETHER SIMILAR ALTERATIONS OF SAM AND SAH LEVELS ARE ALSO PRODUCED IN BRAIN. METHODS: MALE ADULT SPRAGUE DAWLEY RATS WERE FED CHRONICALLY WITH LIEBER-DECARLI ETHANOL (ETOH) (9% V/V) OR CONTROL DIET. THE ETOH-DIET-FED RATS WERE WITHDRAWN FOR 0 AND 24 HOURS. THE CEREBELLUM AND LIVER TISSUES WERE DISSECTED AND USED TO INVESTIGATE CHANGES IN ONE-CARBON METABOLISM, SAM, AND SAH LEVELS. RESULTS: WE FOUND THAT CHRONIC ETOH EXPOSURE DECREASED SAM LEVELS, SAM/SAH RATIO, MS, METHYLENE TETRAHYDROFOLATE REDUCTASE, AND BETAINE HOMOCYSTEINE METHYLTRANSFERASE (BHMT) EXPRESSION AND INCREASED METHIONINE ADENOSYLTRANSFERASE-2B (MAT2B) BUT NOT MAT2A EXPRESSION IN THE LIVER. IN CONTRAST, CHRONIC ETOH EXPOSURE DECREASED SAH LEVELS, INCREASED SAM/SAH RATIO AND THE EXPRESSION OF MAT2A AND S-ADENOSYL HOMOCYSTEINE HYDROLASE, WHILE THE LEVELS OF SAM OR BHMT EXPRESSION IN CEREBELLUM REMAINED UNALTERED. HOWEVER, IN BOTH LIVER AND CEREBELLUM, CHRONIC ETOH EXPOSURE DECREASED THE EXPRESSION OF MS AND INCREASED MAT2B EXPRESSION. ALL CHRONIC ETOH-INDUCED CHANGES OF ONE-CARBON METABOLISM IN CEREBELLUM, BUT NOT LIVER, RETURNED TO NEAR-NORMAL LEVELS DURING ETOH WITHDRAWAL. CONCLUSIONS: THESE RESULTS INDICATE A DECREASED "METHYLATION INDEX" IN LIVER AND AN INCREASED "METHYLATION INDEX" IN CEREBELLUM. THE OPPOSING CHANGES OF THE "METHYLATION INDEX" SUGGEST ALTERED DNA METHYLATION IN LIVER AND CEREBELLUM, THUS IMPLICATING ONE-CARBON METABOLISM IN THE PATHOPHYSIOLOGY OF ALCOHOLISM.	2017	
                                                                                                                                                                                                                                                                                                                                                                                        
7  3619  42 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
8  2792  37 FAT10 IS AN EPIGENETIC MARKER FOR LIVER PRENEOPLASIA IN A DRUG-PRIMED MOUSE MODEL OF TUMORIGENESIS. THERE IS CLINICAL EVIDENCE THAT CHRONIC LIVER DISEASES IN WHICH MDBS (MALLORY DENK BODIES) FORM PROGRESS TO HEPATOCELLULAR CARCINOMA. THE PRESENT STUDY PROVIDES EVIDENCE THAT LINKS MDB FORMATION INDUCED BY CHRONIC DRUG INJURY, WITH PRENEOPLASIA AND LATER TO THE FORMATION OF TUMORS, WHICH DEVELOP LONG AFTER DRUG WITHDRAWAL. EVIDENCE INDICATED THAT THIS LINK WAS DUE TO AN EPIGENETIC CELLULAR MEMORY INDUCED BY CHRONIC DRUG INGESTION. MICROARRAY ANALYSIS SHOWED THAT THE EXPRESSIONS OF MANY MARKERS OF PRENEOPLASIA (UBD, ALPHA FETOPROTEIN, KLF6 AND GLUTATHIONE-S-TRANSFERASE MU2) WERE INCREASED TOGETHER WHEN THE DRUG DDC WAS REFED. THESE CHANGES WERE SUPPRESSED BY S-ADENOSYLMETHIONINE FEEDING, INDICATING THAT THE DRUG WAS AFFECTING DNA AND HISTONES METHYLATION IN AN EPIGENETIC MANNER. THE LINK BETWEEN MDB FORMATION AND NEOPLASIA FORMATION WAS LIKELY DUE TO THE OVER EXPRESSION OF UBD (ALSO CALLED FAT10), WHICH IS UP REGULATED IN 90% OF HUMAN HEPATOCELLULAR CARCINOMAS. IMMUNOHISTOCHEMICAL STAINING OF DRUG-PRIMED MOUSE LIVERS SHOWED THAT FAT10 POSITIVE LIVER CELLS PERSISTED UP TO 4 MONTHS AFTER DRUG WITHDRAWAL AND THEY WERE STILL FOUND IN THE LIVERS OF MICE, 14 MONTHS AFTER DRUG WITHDRAWAL. THE REFEEDING OF DDC INCREASED THE PERCENT OF FAT10 HEPATOCYTES.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
9  6310  46 THE REGULATION OF NON-CODING RNA EXPRESSION IN THE LIVER OF MICE FED DDC. MALLORY-DENK BODIES (MDBS) ARE FOUND IN THE LIVER OF PATIENTS WITH ALCOHOLIC AND CHRONIC NONALCOHOLIC LIVER DISEASE, AND HEPATOCELLULAR CARCINOMA (HCC). DIETHYL 1,4-DIHYDRO-2,4,6,-TRIMETHYL-3,5-PYRIDINEDICARBOXYLATE (DDC) IS USED AS A MODEL TO INDUCE THE FORMATION OF MDBS IN MOUSE LIVER. PREVIOUS STUDIES IN THIS LABORATORY SHOWED THAT DDC INDUCED EPIGENETIC MODIFICATIONS IN DNA AND HISTONES. THE COMBINATION OF THESE MODIFICATIONS CHANGES THE PHENOTYPE OF THE MDB FORMING HEPATOCYTES, AS INDICATED BY THE MARKER FAT10. THESE EPIGENETIC MODIFICATIONS ARE PARTIALLY PREVENTED BY ADDING TO THE DIET S-ADENOSYLMETHIONINE (SAME) OR BETAINE, BOTH METHYL DONORS. THE EXPRESSION OF THREE IMPRINTED NCRNA GENES WAS FOUND TO CHANGE IN MDB FORMING HEPATOCYTES, WHICH IS THE SUBJECT OF THIS REPORT. NCRNA EXPRESSION WAS QUANTITATED BY REAL-TIME PCR AND RNA FISH IN LIVER SECTIONS. MICROARRAY ANALYSIS SHOWED THAT THE EXPRESSION OF THREE NCRNAS WAS REGULATED BY DDC: UP REGULATION OF H19, ANTISENSE IGF2R (AIR), AND DOWN REGULATION OF GTL2 (ALSO CALLED MEG3). S-ADENOSYLMETHIONINE (SAME) FEEDING PREVENTED THESE CHANGES. BETAINE, ANOTHER METHYL GROUP DONOR, PREVENTED ONLY H19 AND AIR UP REGULATION INDUCED BY DDC, ON MICROARRAYS. THE RESULTS OF THE SAME AND BETAINE GROUPS WERE CONFIRMED BY REAL-TIME PCR, EXCEPT FOR AIR EXPRESSION. AFTER 1 MONTH OF DRUG WITHDRAWAL, THE EXPRESSION OF THE THREE NCRNAS TENDED TOWARD CONTROL LEVELS OF EXPRESSION. LIVER TUMORS THAT DEVELOPED ALSO SHOWED UP REGULATION OF H19 AND AIR. THE RNA FISH APPROACH SHOWED THAT THE MDB FORMING CELLS' PHENOTYPE CHANGED THE LEVEL OF EXPRESSION OF AIR, H19 AND GTL2, COMPARED TO THE SURROUNDING CELLS. FURTHERMORE, OVER EXPRESSION OF H19 AND AIR WAS DEMONSTRATED IN TUMORS FORMED IN MICE WITHDRAWN FOR 9 MONTHS. THE DYSREGULATION OF NCRNA IN MDB FORMING LIVER CELLS HAS BEEN OBSERVED FOR THE FIRST TIME IN DRUG-PRIMED MICE ASSOCIATED WITH LIVER PRENEOPLASTIC FOCI AND TUMORS.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                              
10  3841  39 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	
                                                                                                                                                                                                                                                                                                                                                                                                                               
11   286  38 AGING AND ALCOHOL INTERACT TO ALTER HEPATIC DNA HYDROXYMETHYLATION. BACKGROUND: AGING AND CHRONIC ALCOHOL CONSUMPTION ARE BOTH MODIFIERS OF DNA METHYLATION, BUT IT IS NOT YET KNOWN WHETHER CHRONIC ALCOHOL CONSUMPTION ALSO ALTERS DNA HYDROXYMETHYLATION, A NEWLY DISCOVERED EPIGENETIC MARK PRODUCED BY OXIDATION OF METHYLCYTOSINE. FURTHERMORE, IT HAS NOT BEEN TESTED WHETHER AGING AND ALCOHOL INTERACT TO MODIFY THIS EPIGENETIC PHENOMENON, THEREBY HAVING AN INDEPENDENT EFFECT ON GENE EXPRESSION. METHODS: OLD (18 MONTHS) AND YOUNG (4 MONTHS) MALE C57BL/6 MICE WERE PAIR-FED EITHER A LIEBER-DECARLI LIQUID DIET WITH ALCOHOL (18% OF ENERGY) OR AN ISOCALORIC LIEBER-DECARLI CONTROL DIET FOR 5 WEEKS. GLOBAL DNA HYDROXYMETHYLATION AND DNA METHYLATION WERE ANALYZED FROM HEPATIC DNA USING A NEW LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY METHOD. HEPATIC MRNA EXPRESSION OF THE TET ENZYMES WERE MEASURED VIA QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION. RESULTS: IN YOUNG MICE, MILD CHRONIC ALCOHOL EXPOSURE SIGNIFICANTLY REDUCED GLOBAL DNA HYDROXYMETHYLATION COMPARED WITH CONTROL MICE (0.22 +/- 0.01 VS. 0.29 +/- 0.06%, P = 0.004). ALCOHOL DID NOT SIGNIFICANTLY ALTER HYDROXYMETHYLCYTOSINE LEVELS IN OLD MICE. OLD MICE FED THE CONTROL DIET SHOWED DECREASED GLOBAL DNA HYDROXYMETHYLATION COMPARED WITH YOUNG MICE FED THE CONTROL DIET (0.24 +/- 0.02 VS. 0.29 +/- 0.06%, P = 0.04). THIS MODEL SUGGESTS AN INTERACTION BETWEEN AGING AND ALCOHOL IN DETERMINING DNA HYDROXYMETHYLATION (PINTERACTION = 0.009). EXPRESSION OF TET2 AND TET3 WAS DECREASED IN THE OLD MICE RELATIVE TO THE YOUNG (P < 0.005). CONCLUSIONS: THE OBSERVATION THAT ALCOHOL ALTERS DNA HYDROXYMETHYLATION INDICATES A NEW EPIGENETIC EFFECT OF ALCOHOL. THIS IS THE FIRST STUDY DEMONSTRATING THE INTERACTIVE EFFECTS OF CHRONIC ALCOHOL CONSUMPTION AND AGING ON DNA HYDROXYMETHYLATION.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
12  2156  49 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	
                                                                                                                                                                                                                                                                                                      
13  3081  34 GENOME-WIDE PROMOTER DNA METHYLATION PROFILING OF HEPATOCELLULAR CARCINOMAS ARISING EITHER SPONTANEOUSLY OR DUE TO CHRONIC EXPOSURE TO GINKGO BILOBA EXTRACT (GBE) IN B6C3F1/N MICE. EPIGENETIC MODIFICATIONS, SUCH AS DNA METHYLATION, PLAY AN IMPORTANT ROLE IN CARCINOGENESIS. IN A RECENT NTP STUDY, CHRONIC EXPOSURE OF B6C3F1/N MICE TO GINKGO BILOBA EXTRACT (GBE) RESULTED IN A HIGH INCIDENCE OF HEPATOCELLULAR CARCINOMAS (HCC). GENOME-WIDE PROMOTER METHYLATION PROFILING ON GBE-EXPOSED HCC (2000 MG/KG GROUP), SPONTANEOUS HCC (VEHICLE-CONTROL GROUP), AND AGE-MATCHED VEHICLE CONTROL LIVER WAS PERFORMED TO IDENTIFY DIFFERENTIALLY METHYLATED GENES IN GBE-EXPOSED HCC AND SPONTANEOUS HCC. DNA METHYLATION ALTERATIONS WERE CORRELATED TO THE CORRESPONDING GLOBAL GENE EXPRESSION CHANGES. COMPARED TO CONTROL LIVER, 1296 GENE PROMOTERS (719 HYPERMETHYLATED, 577 HYPOMETHYLATED) IN GBE-EXPOSED HCC AND 738 (427 HYPERMETHYLATED, 311 HYPOMETHYLATED) GENE PROMOTERS IN SPONTANEOUS HCC WERE SIGNIFICANTLY DIFFERENTIALLY METHYLATED, SUGGESTING AN IMPACT OF METHYLATION ON GBE-EXPOSED HCC. DIFFERENTIAL METHYLATION OF PROMOTER REGIONS IN RELEVANT CANCER GENES (CMYC, SPRY2, DUSP5) AND THEIR CORRESPONDING DIFFERENTIAL GENE EXPRESSION WAS VALIDATED BY QUANTITATIVE PYROSEQUENCING AND QRT-PCR, RESPECTIVELY. IN CONCLUSION, WE HAVE IDENTIFIED DIFFERENTIALLY METHYLATED PROMOTER REGIONS OF RELEVANT CANCER GENES ALTERED IN GBE-EXPOSED HCC COMPARED TO SPONTANEOUS HCC. FURTHER STUDY OF UNIQUE SETS OF DIFFERENTIALLY METHYLATED GENES IN CHEMICAL-EXPOSED MOUSE HCC COULD POTENTIALLY BE USED TO DIFFERENTIATE TREATMENT-RELATED TUMORS FROM SPONTANEOUS-TUMORS IN CANCER BIOASSAYS AND PROVIDE ADDITIONAL UNDERSTANDING OF THE UNDERLYING EPIGENETIC MECHANISMS OF CHEMICAL CARCINOGENESIS.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
14  4581  32 N-METHYL-D-ASPARTATE 2B RECEPTOR SUBTYPE (NR2B) PROMOTER METHYLATION IN PATIENTS DURING ALCOHOL WITHDRAWAL. NMDA RECEPTORS AND ESPECIALLY THE NR2B RECEPTOR SUBTYPE PLAY A CRUCIAL ROLE DURING CHRONIC ETHANOL CONSUMPTION AND ALCOHOL WITHDRAWAL. THEREFORE, THE NR2B RECEPTOR SUBTYPE EXPRESSION IN PERIPHERAL BLOOD CELLS OF 32 MALE PATIENTS SUFFERING FROM ALCOHOL DEPENDENCY WERE ASSESSED THROUGH QUANTITATIVE RT-PCR AND TO EXPLORE REGULATING EPIGENETIC MECHANISMS, A METHYLATION ANALYSIS WAS CONDUCTED USING BISULFITE SEQUENCING OF A FRAGMENT OF THE NR2B PROMOTER REGION. THE EXPRESSION OF THE NR2B RECEPTOR INCREASED SIGNIFICANTLY DURING THE FIRST 24 H OF WITHDRAWAL TREATMENT (DAY 1; T = 4.1, P = 0.001), AND ALSO ON AND DAY 3 (T = 2.4; P = 0.029). THE SEVERITY OF ALCOHOL DRINKING PATTERN, MEASURED BY LIFETIME DRINKING AND DAILY ETHANOL INTAKE, WAS NEGATIVELY CORRELATED WITH THE METHYLATION OF A DEFINED CLUSTER OF FIVE CPG-SITES WITHIN THE NR2B PROMOTER (LIFETIME DRINKING: SPEARMAN'S RHO = -0.55; P = 0.013; DAILY ETHANOL INTAKE: RHO = -0.46; P = 0.043). THESE FINDINGS MIGHT EXPLAIN THE OBSERVATION OF AN IMPACT OF ALCOHOL CONSUMPTION PATTERNS ON THE GRAVITY OF WITHDRAWAL SYMPTOMS.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
15   219  38 ACUTE IMMOBILIZATION STRESS FOLLOWING CONTEXTUAL FEAR CONDITIONING REDUCES FEAR MEMORY: TIMING IS ESSENTIAL. BACKGROUND: HISTONE ACETYLATION IS REGULATED IN RESPONSE TO STRESS AND PLAYS AN IMPORTANT ROLE IN LEARNING AND MEMORY. CHRONIC STRESS IS KNOWN TO DETERIORATE COGNITION, WHEREAS ACUTE STRESS FACILITATES MEMORY FORMATION. HOWEVER, WHETHER ACUTE STRESS FACILITATES MEMORY FORMATION WHEN IT IS APPLIED AFTER FEAR STIMULATION IS NOT YET KNOWN. THEREFORE, THIS STUDY AIMED TO INVESTIGATE THE EFFECT OF ACUTE STRESS APPLIED AFTER FEAR TRAINING ON MEMORY FORMATION, MRNA EXPRESSION OF BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF), EPIGENETIC REGULATION OF BDNF EXPRESSION, AND CORTICOSTERONE LEVEL IN MICE IN VIVO. METHODS: MICE WERE SUBJECTED TO ACUTE IMMOBILIZATION STRESS FOR 30 MIN AT 60 OR 90 MIN AFTER CONTEXTUAL FEAR CONDITIONING TRAINING, AND ACETYLATION OF HISTONE 3 AT LYSINE 14 (H3K14) AND LEVEL OF CORTICOSTERONE WERE MEASURED USING WESTERN BLOT ANALYSIS AND ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA), RESPECTIVELY. A FREEZING BEHAVIOR TEST WAS PERFORMED 24 H AFTER TRAINING, AND MRNA EXPRESSION OF BDNF WAS MEASURED USING REAL-TIME POLYMERASE CHAIN REACTIONS. DIFFERENT GROUPS OF MICE WERE USED FOR EACH TEST. RESULTS: FREEZING BEHAVIOR SIGNIFICANTLY DECREASED WITH THE DOWN-REGULATION OF BDNF MRNA EXPRESSION CAUSED BY ACUTE IMMOBILIZATION STRESS AT 60 MIN AFTER FEAR CONDITIONING TRAINING OWING TO THE REDUCTION OF H3K14 ACETYLATION. HOWEVER, BDNF MRNA EXPRESSION AND H3K14 ACETYLATION WERE NOT REDUCED IN ANIMALS SUBJECTED TO IMMOBILIZATION STRESS AT 90 MIN AFTER THE TRAINING. FURTHER, THE CORTICOSTERONE LEVEL WAS SIGNIFICANTLY HIGH IN MICE SUBJECTED TO IMMOBILIZATION STRESS AT 60 MIN AFTER THE TRAINING. CONCLUSION: ACUTE IMMOBILIZATION STRESS FOR 30 MIN AT 60 MIN AFTER FEAR CONDITIONING TRAINING IMPAIRED MEMORY FORMATION AND REDUCED BDNF MRNA EXPRESSION AND H3K14 ACETYLATION IN THE HIPPOCAMPUS OF MICE OWING TO THE HIGH LEVEL OF CORTICOSTERONE.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
16  5791  45 STABLE HISTONE METHYLATION CHANGES AT PROTEOGLYCAN NETWORK GENES FOLLOWING ETHANOL EXPOSURE. ALCOHOL USE DISORDER (AUD) IS A CHRONIC MENTAL ILLNESS IN WHICH PATIENTS OFTEN ACHIEVE PROTRACTED PERIODS OF ABSTINENCE PRIOR TO RELAPSE. EPIGENETIC MECHANISMS MAY PROVIDE AN EXPLANATION FOR THE PERSISTING GENE EXPRESSION CHANGES THAT CAN BE OBSERVED EVEN AFTER LONG PERIODS OF ABSTINENCE AND MAY CONTRIBUTE TO RELAPSE. IN THIS STUDY, WE EXAMINED TWO HISTONE MODIFICATIONS, HISTONE 3 LYSINE 4 TRI-METHYLATION (H3K4ME3) AND HISTONE 3 LYSINE 27 TRI-METHYLATION (H3K27ME3), IN THE PREFRONTAL CORTEX OF WITHDRAWAL SEIZURE RESISTANT (WSR) MICE 21 DAYS AFTER 72 H OF ETHANOL VAPOR EXPOSURE. THESE HISTONE MODIFICATIONS WERE SELECTED BECAUSE THEY ARE ASSOCIATED WITH ACTIVE PROMOTERS (H3K4ME3) AND REPRESSED GENE EXPRESSION IN A EUCHROMATIC ENVIRONMENT (H3K27ME3). WE PERFORMED A GENOME-WIDE ANALYSIS TO IDENTIFY DIFFERENCES IN H3K4ME3 AND H3K27ME3 LEVELS IN POST-ETHANOL EXPOSURE VS. CONTROL MICE BY CHIP-SEQ. WE DETECTED A GLOBAL REDUCTION IN H3K4ME3 PEAKS AND INCREASE IN H3K27ME3 PEAKS IN POST-ETHANOL EXPOSURE MICE COMPARED TO CONTROLS, THESE CHANGES ARE CONSISTENT WITH PERSISTENT REDUCTIONS IN GENE EXPRESSION. PATHWAY ANALYSIS OF GENES DISPLAYING CHANGES IN H3K4ME3 AND H3K27ME3 REVEALED ENRICHMENT FOR GENES INVOLVED IN PROTEOGLYCAN AND CALCIUM SIGNALING PATHWAYS, RESPECTIVELY. MICROARRAY ANALYSIS OF 7,683 GENES AND QPCR ANALYSIS IDENTIFIED EIGHT GENES DISPLAYING CONCORDANT REGULATION OF GENE EXPRESSION AND H3K4ME3/H3K27ME3. WE ALSO COMPARED CHANGES IN H3K4ME3 AND/OR H3K27ME3 FROM OUR STUDY WITH CHANGES IN GENE EXPRESSION IN RESPONSE TO ETHANOL FROM PUBLISHED LITERATURE AND WE FOUND THAT THE EXPRESSION OF 52% OF THE GENES WITH ALTERED H3K4ME3 BINDING AND 40% OF GENES WITH H3K27ME3 DIFFERENCES ARE ALTERED BY ETHANOL EXPOSURE. THE CHROMATIN CHANGES ASSOCIATED WITH THE 21-DAY POST-EXPOSURE PERIOD SUGGEST THAT THIS PERIOD IS A UNIQUE STATE IN THE ADDICTION CYCLE THAT DIFFERS FROM ETHANOL INTOXICATION AND ACUTE WITHDRAWAL. THESE RESULTS PROVIDE INSIGHTS INTO THE ENDURING EFFECTS OF ETHANOL ON PROTEOGLYCAN AND CALCIUM SIGNALING GENES IN THE BRAIN.	2018	
                                                                                                                                                                                                                                                                                                               
17  2347  42 EPIGENETIC REGULATION OF MIR-124 UNDER ETHANOL DEPENDENCE AND WITHDRAWAL. WITHDRAWAL FROM CHRONIC ALCOHOL CAUSE THE PERSISTENT MOLECULAR ALTERATION, SUCH AS CHANGES IN THE RELEASE OF NEUROTRANSMITTER AND GENE EXPRESSION. THE ALTERATIONS ARE THOUGHT TO INCREASE IN THE RISK OF RELAPSE. RECENT STUDIES SUGGEST THAT THE GENE EXPRESSION REGULATED BY HISTONE ACETYLATION MAY PLAY AN IMPORTANT ROLE IN THE DEPENDENCE OF ABUSED DRUGS, INCLUDING OF ETHANOL. FURTHERMORE, MIRNA, ANOTHER REGULATOR OF GENE EXPRESSION, ARE ALSO IMPORTANT MOLECULES FOR THE DEPENDENCE. HOWEVER, CHANGES IN THE MOLECULES UNDER ETHANOL WITHDRAWAL AND ITS RELATIONSHIP ARE POORLY UNDERSTOOD. IN THE PRESENT STUDY, WE INVESTIGATED THE EXPRESSION OF ACETYLATED HISTONE H3 AND MIR-124 IN MOUSE BRAIN AT 3 DAYS AFTER ETHANOL WITHDRAWAL. 6-WEEK AGES OF C57BL/6J MICE WERE TREATED WITH LIQUID DIET CONTAINING ETHANOL FOR 10 DAYS. USING THE ESCALATING ETHANOL DOSAGE SCHEDULE, THE MICE WERE FED THE ETHANOL DIET AS FOLLOWS: 1ST DAY: 1 W/V%: 2ND AND 3RD DAY: 3 W/V%; 4TH AND 5TH DAY: 4 W/V% AND FROM THE 6TH TO 10TH DAY: 5 W/V% ETHANOL DIET, RESPECTIVELY. THE PAIR-FED CONTROL MICE WERE GIVEN THE SAME VOLUME OF ETHANOL-FREE LIQUID DIET WITH GLUCOSE SUBSTITUTED IN ISOCALORIC QUANTITIES FOR ETHANOL. AFTER FEEDING ALCOHOL LIQUID DIET, THE MICE SHOWED SEVERE WITHDRAWAL SIGNS. THE EXPRESSION OF ACETYLATED HISTONE H3 WAS SIGNIFICANTLY DECREASED IN LIMBIC FOREBRAIN AT 3 DAYS AFTER WITHDRAWAL. WE FOUND THAT MIR-124 ALSO DECREASED IN THE LIMBIC FOREBRAIN. IT HAS BEEN REPORTED THAT CDC42 REGULATES NEURONAL DEVELOPMENT AS A TARGET OF MIR-124. WE FOUND THAT CDC42 PROTEIN MARKEDLY INCREASED IN BOTH BRAIN REGIONS AT 3 DAYS AFTER WITHDRAWAL. OUR FINDINGS SUGGEST THAT CHANGES IN THE EXPRESSION OF MIR-124 VIA HISTONE ACETYLATION LEADS TO CHANGE THE CDC42 EXPRESSION UNDER ETHANOL WITHDRAWAL.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
18  4930  34 PATERNAL ALCOHOL EXPOSURE REDUCES ACQUISITION OF OPERANT ALCOHOL SELF-ADMINISTRATION AND AFFECTS BDNF DNA METHYLATION IN MALE AND FEMALE OFFSPRING. FAMILIAL TRANSMISSION OF ALCOHOL USE DISORDER REFLECTS GENETIC AND ENVIRONMENTAL FACTORS. PATERNAL ALCOHOL EXPOSURE MAY AFFECT RODENT OFFSPRING VIA EPIGENETIC MODIFICATIONS TRANSMITTED THROUGH THE MALE GERM LINE. WHILE SUCH EXPOSURE ALTERS ALCOHOL SENSITIVITY IN MOUSE OFFSPRING, NO STUDIES EXAMINED IF IT IMPACTS THE DEVELOPMENT OF OPERANT ALCOHOL SELF-ADMINISTRATION IN RATS. WE EXPOSED MALE (SIRES) WISTAR RATS TO CHRONIC INTERMITTENT ETHANOL IN VAPOUR CHAMBERS (16 H/DAY; 5 DAYS/WEEK) OR TO AIR FOR 6 WEEKS. EIGHT WEEKS LATER, RATS WERE MATED WITH ALCOHOL-NAIVE FEMALES. ADULT ALCOHOL- AND CONTROL-SIRED F1 OFFSPRING WERE ASSESSED IN ACQUISITION OF ALCOHOL SELF-ADMINISTRATION IN WHICH INCREASING ALCOHOL CONCENTRATIONS (2.5%, 5% AND 10%, V/V) WERE DELIVERED AFTER ONE LEVER PRESS (FIXED RATIO 1 OR FR1). PRIOR TO ALCOHOL SESSIONS, RATS WERE TRAINED TO LEVER PRESS FOR FOOD DELIVERY UNDER AN FR1 SCHEDULE OF REINFORCEMENT. DNA METHYLATION LEVELS OF THE BRAIN DERIVED NEUROTROPHIC FACTOR (BDNF) GENE WERE MEASURED IN SPERM, NUCLEUS ACCUMBENS (NAC) AND MEDIAL PREFRONTAL CORTEX (MPFC) IN SIRES AND IN OFFSPRING. ALCOHOL-EXPOSED SIRES HAD LOWER BDNF DNA METHYLATION LEVELS IN NAC AND GREATER METHYLATION LEVELS IN MPFC. ALTHOUGH THIS PATTERN WAS NOT RECAPITULATED IN OFFSPRING, ALCOHOL-SIRED OFFSPRING OF BOTH SEXES DID SHOW ABERRANT BDNF DNA METHYLATION PATTERNS COMPARED TO CONTROL-SIRED OFFSPRING. ALCOHOL-SIRED OFFSPRING SELF-ADMINISTERED LESS ALCOHOL (5% AND 10%) WITH NO GROUP DIFFERENCES IN FOOD RESPONDING. RESULTS INDICATE THAT PATERNAL ALCOHOL EXPOSURE PRIOR TO CONCEPTION PROTECTS AGAINST ALCOHOL'S INITIAL REINFORCING EFFECTS BUT THE PATTERN OF DYSREGULATED BDNF METHYLATION IN REWARD-RELATED CIRCUITRY DID NOT MIMIC CHANGES SEEN IN SIRES.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
19  1800  27 EFFECT OF HISTONE DEACETYLASE INHIBITOR ON ETHANOL WITHDRAWAL-INDUCED HYPERALGESIA IN RATS. BACKGROUND: INCREASED PAIN SENSITIVITY IS OBSERVED FOLLOWING ALCOHOL WITHDRAWAL, AND ATTEMPTS TO ALLEVIATE THIS HYPERALGESIA CAN CONTRIBUTE TO THE CYCLE OF ADDICTION. THE AIM OF THIS STUDY WAS TO DETERMINE IF ALCOHOL WITHDRAWAL-INDUCED HYPERALGESIA WAS OBSERVED IN A CHRONIC ETHANOL EXPOSURE MODEL AND IF THIS PAIN WAS AFFECTED BY HISTONE DEACETYLASE INHIBITORS, THUS REVEALING AN EPIGENETIC MECHANISM. METHODS: ADULT MALE SPRAGUE DAWLEY RATS RECEIVED LIEBER-DECARLI LIQUID CONTROL OR ETHANOL (9% V/V) DIET FOR 15 DAYS. MECHANICAL SENSITIVITY WAS MEASURED WITH VON FREY HAIR STIMULATION OF THE HINDPAW DURING ETHANOL ADMINISTRATION AND 24- AND 72-HOUR WITHDRAWAL. RESULTS: ETHANOL WITHDRAWAL PRODUCED SEVERE AND SUSTAINED MECHANICAL HYPERALGESIA, AN EFFECT NOT OBSERVED IN THE CONTROL OR ETHANOL-MAINTAINED GROUPS. FURTHERMORE, THIS HYPERALGESIA WAS ATTENUATED BY THE HISTONE DEACETYLASE INHIBITOR, SUBEROYLANILIDE HYDROXAMIC ACID TREATMENT. CONCLUSIONS: HEIGHTENED PAIN SENSITIVITY WAS OBSERVED FOLLOWING WITHDRAWAL FROM CHRONIC ETHANOL EXPOSURE, AND HISTONE DEACETYLASE INHIBITORS COULD BE NOVEL TREATMENTS FOR THIS ALCOHOL WITHDRAWAL-INDUCED HYPERALGESIA.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
20  4943  31 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