1 1429 150 DIFFERENTIAL EXPRESSION OF MICRORNAS IN THE HIPPOCAMPI OF MALE AND FEMALE RODENTS AFTER CHRONIC ALCOHOL ADMINISTRATION. BACKGROUND: WOMEN ARE MORE VULNERABLE THAN MEN TO THE NEUROTOXICITY AND SEVERE BRAIN DAMAGE CAUSED BY CHRONIC HEAVY ALCOHOL USE. IN ADDITION, BRAIN DAMAGE DUE TO CHRONIC HEAVY ALCOHOL USE MAY BE ASSOCIATED WITH SEX-DEPENDENT EPIGENETIC MODIFICATIONS. THIS STUDY AIMED TO IDENTIFY MICRORNAS (MIRNAS) AND THEIR TARGET GENES THAT ARE DIFFERENTIALLY EXPRESSED IN THE HIPPOCAMPI OF MALE AND FEMALE ANIMAL MODELS IN RESPONSE TO ALCOHOL. METHODS: AFTER CHRONIC ALCOHOL ADMINISTRATION (3~3.5 G/KG/DAY) IN MALE (CONTROL, N = 10; ALCOHOL, N = 12) OR FEMALE (CONTROL, N = 10; ALCOHOL, N = 12) SPRAGUE-DAWLEY RATS FOR 6 WEEKS, WE MEASURED BODY WEIGHTS AND DOUBLECORTIN (DCX; A NEUROGENESIS MARKER) CONCENTRATIONS AND ANALYZED UP- OR DOWNREGULATED MIRNAS USING GENECHIP MIRNA 4.0 ARRAYS. THE DIFFERENTIALLY EXPRESSED MIRNAS AND THEIR PUTATIVE TARGET GENES WERE VALIDATED BY RT-QPCR. RESULTS: ALCOHOL ATTENUATED BODY WEIGHT GAIN ONLY IN THE MALE GROUP. ON THE OTHER HAND, ALCOHOL LED TO INCREASED SERUM AST IN FEMALE RATS AND DECREASED SERUM TOTAL CHOLESTEROL CONCENTRATIONS IN MALE RATS. THE EXPRESSION OF DCX WAS SIGNIFICANTLY REDUCED IN THE HIPPOCAMPI OF MALE ALCOHOL-TREATED RATS. NINE MIRNAS WERE SIGNIFICANTLY UP- OR DOWNREGULATED IN MALE ALCOHOL-TREATED RATS, INCLUDING UPREGULATION OF MIR-125A-3P, LET-7A-5P, AND MIR-3541, AND DOWNREGULATION OF THEIR TARGET GENES (PRDM5, SUV39H1, PTPRZ1, MAPK9, ING4, WT1, NKX3-1, DAB2IP, RNF152, RIPK1, LIN28A, APBB3, NRAS, AND ACVR1C). ON THE OTHER HAND, 7 MIRNAS WERE SIGNIFICANTLY UP- OR DOWNREGULATED IN ALCOHOL-TREATED FEMALE RATS, INCLUDING DOWNREGULATION OF MIR-881-3P AND MIR-504 AND UPREGULATION OF THEIR TARGET GENES (NAA50, CLOCK, CBFB, ARIH1, UBE2G1, AND GNG7). CONCLUSIONS: THESE RESULTS SUGGEST THAT CHRONIC HEAVY ALCOHOL USE PRODUCES SEX-DEPENDENT EFFECTS ON NEUROGENESIS AND MIRNA EXPRESSION IN THE HIPPOCAMPUS AND THAT SEX DIFFERENCES SHOULD BE CONSIDERED WHEN DEVELOPING MIRNA BIOMARKERS TO DIAGNOSE OR TREAT ALCOHOLICS.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
2 2674  35 ETHOSUXIMIDE REDUCES EPILEPTOGENESIS AND BEHAVIORAL COMORBIDITY IN THE GAERS MODEL OF GENETIC GENERALIZED EPILEPSY. PURPOSE: ETHOSUXIMIDE (ESX) IS A DRUG OF CHOICE FOR THE SYMPTOMATIC TREATMENT OF ABSENCE SEIZURES. CHRONIC TREATMENT WITH ESX HAS BEEN REPORTED TO HAVE DISEASE-MODIFYING ANTIEPILEPTOGENIC ACTIVITY IN THE WAG/RIJ RAT MODEL OF GENETIC GENERALIZED EPILEPSY (GGE) WITH ABSENCE SEIZURES. HERE WE EXAMINED WHETHER CHRONIC TREATMENT WITH ESX (1) POSSESSES ANTIEPILEPTOGENIC EFFECTS IN THE GENETIC ABSENCE EPILEPSY RATS FROM STRASBOURG (GAERS) MODEL OF GGE, (2) IS ASSOCIATED WITH A MITIGATION OF BEHAVIORAL COMORBIDITIES, AND (3) INFLUENCES GENE EXPRESSION IN THE SOMATOSENSORY CORTEX REGION WHERE SEIZURES ARE THOUGHT TO ORIGINATE. METHODS: GAERS AND NONEPILEPTIC CONTROL (NEC) RATS WERE CHRONICALLY TREATED WITH ESX (IN DRINKING WATER) OR CONTROL (TAP WATER) FROM 3 TO 22 WEEKS OF AGE. SUBSEQUENTLY, ALL ANIMALS RECEIVED TAP WATER ONLY FOR ANOTHER 12 WEEKS TO ASSESS ENDURING EFFECTS OF TREATMENT. SEIZURE FREQUENCY AND ANXIETY-LIKE BEHAVIORS WERE SERIALLY ASSESSED THROUGHOUT THE EXPERIMENTAL PARADIGM. TREATMENT EFFECTS ON THE EXPRESSION OF KEY COMPONENTS OF THE EPIGENETIC MOLECULAR MACHINERY, THE DNA METHYLTRANSFERASE ENZYMES, WERE ASSESSED USING QUANTITATIVE POLYMERASE CHAIN REACTION (QPCR). KEY FINDINGS: ESX TREATMENT SIGNIFICANTLY REDUCED SEIZURES IN GAERS DURING THE TREATMENT PHASE, AND THIS EFFECT WAS MAINTAINED DURING THE 12-WEEK POSTTREATMENT PHASE (P < 0.05). FURTHERMORE, THE ANXIETY-LIKE BEHAVIORS PRESENT IN GAERS WERE REDUCED BY ESX TREATMENT (P < 0.05). MOLECULAR ANALYSIS REVEALED THAT ESX TREATMENT WAS ASSOCIATED WITH INCREASED EXPRESSION OF DNA METHYLTRANSFERASE ENZYME MESSENGER RNA (MRNA) IN CORTEX. SIGNIFICANCE: CHRONIC ESX TREATMENT HAS DISEASE-MODIFYING EFFECTS IN THE GAERS MODEL OF GGE, WITH ANTIEPILEPTOGENIC EFFECTS AGAINST ABSENCE SEIZURES AND MITIGATION OF BEHAVIORAL COMORBIDITIES. THE CELLULAR MECHANISM FOR THESE EFFECTS MAY INVOLVE EPIGENETIC MODIFICATIONS.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
3 2978  41 GENETIC ASSOCIATION AND EXPRESSION ANALYSES OF THE PHOSPHATIDYLINOSITOL-4-PHOSPHATE 5-KINASE (PIP5K1C) GENE IN ALCOHOL USE DISORDER-RELEVANCE FOR PAIN SIGNALING AND ALCOHOL USE. BACKGROUND: THE GENE ENCODING PHOSPHATIDYLINOSITOL-4-PHOSPHATE 5-KINASE (PIP5K1C) HAS BEEN RECENTLY IMPLICATED IN PAIN REGULATION. INTERESTINGLY, A RECENT CROSS-TISSUE AND CROSS-PHENOTYPIC EPIGENETIC ANALYSIS IDENTIFIED THE SAME GENE IN ALCOHOL USE DISORDER (AUD). GIVEN THE HIGH COMORBIDITY BETWEEN AUD AND CHRONIC PAIN, WE HYPOTHESIZED THAT GENETIC VARIATION IN PIP5K1C MIGHT CONTRIBUTE TO SUSCEPTIBILITY TO AUD. METHODS: WE CONDUCTED A CASE-CONTROL ASSOCIATION STUDY OF GENETIC VARIANTS IN PIP5K1C. ASSOCIATION ANALYSES OF 16 COMMON PIP5K1C SINGLE NUCLEOTIDE POLYMORPHISMS (SNPS) WERE CONDUCTED IN CASES AND CONTROLS OF AFRICAN (427 CASES AND 137 CONTROLS) AND EUROPEAN ANCESTRY (488 CASES AND 324 CONTROLS) USING STANDARD METHODS. IN ADDITION, GIVEN THE PROMINENT ROLE OF THE OPIOID SYSTEM IN PAIN SIGNALING, WE INVESTIGATED THE EFFECTS OF ACUTE ALCOHOL EXPOSURE ON PIP5K1C EXPRESSION IN HUMANIZED TRANSGENIC MICE FOR THE MU-OPIOID RECEPTOR THAT INCLUDED THE OPRM1 A118G POLYMORPHISM, A WIDELY USED MOUSE MODEL TO STUDY ANALGESIC RESPONSE TO OPIOIDS IN PAIN. PIP5K1C EXPRESSION WAS MEASURED IN THE THALAMUS AND BASOLATERAL AMYGDALA (BLA) IN MICE AFTER SHORT-TERM ADMINISTRATION (SINGLE 2 G/KG DOSE) OF ALCOHOL OR SALINE USING IMMUNOHISTOCHEMISTRY AND ANALYZED BY 2-WAY ANALYSIS OF VARIANCE. RESULTS: IN THE CASE-CONTROL ASSOCIATION STUDY USING AN NIAAA DISCOVERY SAMPLE, 8 SNPS IN PIP5K1C WERE SIGNIFICANTLY ASSOCIATED WITH AUD IN THE AFRICAN ANCESTRY (AA) GROUP (P < 0.05 AFTER CORRECTION; RS4807493, RS10405681, RS2074957, RS10432303, RS8109485, RS1476592, RS10419980, AND RS4432372). HOWEVER, A REPLICATION ANALYSIS USING AN INDEPENDENT SAMPLE (N = 3,801) FOUND NO SIGNIFICANT ASSOCIATIONS AFTER CORRECTION FOR MULTIPLE TESTING. IN THE HUMANIZED TRANSGENIC MOUSE MODEL WITH THE OPRM1 POLYMORPHISM, PIP5K1C EXPRESSION WAS SIGNIFICANTLY DIFFERENT BETWEEN ALCOHOL AND SALINE-TREATED MICE, REGARDLESS OF GENOTYPE, IN BOTH THE THALAMUS (P < 0.05) AND BLA (P < 0.01). CONCLUSIONS: OUR DISCOVERY SAMPLE SHOWS THAT GENETIC VARIANTS IN PIP5K1C ARE ASSOCIATED WITH AUD IN THE AA GROUP, AND ACUTE ALCOHOL EXPOSURE LEADS TO UP-REGULATION OF PIP5K1C, POTENTIALLY EXPLAINING A MECHANISM UNDERLYING THE INCREASED RISK FOR CHRONIC PAIN CONDITIONS IN INDIVIDUALS WITH AUD.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
4 3494  39 IDENTIFICATION OF MICRORNAS WITH DYSREGULATED EXPRESSION IN STATUS EPILEPTICUS INDUCED EPILEPTOGENESIS. THE INVOLVEMENT OF MIRNA IN MESIAL TEMPORAL LOBE EPILEPSY (MTLE) PATHOGENESIS HAS INCREASINGLY BECOME A FOCUS OF EPIGENETIC STUDIES. DESPITE ADVANCES, THE NUMBER OF KNOWN MIRNAS WITH A CONSISTENT EXPRESSION RESPONSE DURING EPILEPTOGENESIS IS STILL SMALL. ADDRESSING THIS SITUATION REQUIRES ADDITIONAL MIRNA PROFILING STUDIES COUPLED TO DETAILED INDIVIDUAL EXPRESSION ANALYSES. HERE, WE PERFORM A MIRNA MICROARRAY ANALYSIS OF THE HIPPOCAMPUS OF WISTAR RATS 24 HOURS AFTER INTRA-HIPPOCAMPAL PILOCARPINE-INDUCED STATUS EPILEPTICUS (H-PILO SE). WE IDENTIFIED 73 MIRNAS THAT UNDERGO SIGNIFICANT CHANGES, OF WHICH 36 WERE UP-REGULATED AND 37 WERE DOWN-REGULATED. TO VALIDATE, WE SELECTED 5 OF THESE (10A-5P, 128A-3P, 196B-5P, 352 AND 324-3P) FOR RT-QPCR ANALYSIS. OUR RESULTS CONFIRMED THAT MIR-352 AND 196B-5P LEVELS WERE SIGNIFICANTLY HIGHER AND MIR-128A-3P LEVELS WERE SIGNIFICANTLY LOWER IN THE HIPPOCAMPUS OF H-PILO SE RATS. WE ALSO EVALUATED WHETHER THE 3 MIRNAS SHOW A DYSREGULATED HIPPOCAMPAL EXPRESSION AT THREE TIME PERIODS (0H, 24H AND CHRONIC PHASE) AFTER SYSTEMIC PILOCARPINE-INDUCED STATUS EPILEPTICUS (S-PILO SE). WE DEMONSTRATE THAT MIR-128A-3P TRANSCRIPTS ARE SIGNIFICANTLY REDUCED AT ALL TIME POINTS COMPARED TO THE NAIVE GROUP. MOREOVER, MIR-196B-5P WAS SIGNIFICANTLY HIGHER ONLY AT 24H POST-SE, WHILE MIR-352 TRANSCRIPTS WERE SIGNIFICANTLY UP-REGULATED AFTER 24H AND IN CHRONIC PHASE (EPILEPTIC) RATS. FINALLY, WHEN WE COMPARED HIPPOCAMPI OF EPILEPTIC AND NON-EPILEPTIC HUMANS, WE OBSERVED THAT TRANSCRIPT LEVELS OF MIRNAS SHOW SIMILAR TRENDS TO THE ANIMAL MODELS. IN SUMMARY, WE SUCCESSFULLY IDENTIFIED TWO NOVEL DYSREGULATED MIRNAS (196B-5P AND 352) AND CONFIRMED MIR-128A-3P DOWNREGULATION IN SE-INDUCED EPILEPTOGENESIS. FURTHER FUNCTIONAL ASSAYS ARE REQUIRED TO UNDERSTAND THE ROLE OF THESE MIRNAS IN MTLE PATHOGENESIS.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
5 1622  35 DNA METHYLTRANSFERASES IN MALAR MELASMA AND THEIR MODIFICATION BY SUNSCREEN IN COMBINATION WITH 4% NIACINAMIDE, 0.05% RETINOIC ACID, OR PLACEBO. BACKGROUND: MALAR MELASMA HAS A CHRONIC AND RECURRENT CHARACTER THAT MAY BE RELATED TO EPIGENETIC CHANGES. OBJECTIVE: TO RECOGNIZE THE EXPRESSION AND DNA METHYLATION OF DNA METHYLTRANSFERASES (DNMTS) IN MALAR MELASMA AND PERILESIONAL SKIN, AS WELL AS THE CHANGES IN DNMTS AFTER THEIR TREATMENT WITH SUNSCREEN IN COMBINATION WITH 4% NIACINAMIDE, 0.05% RETINOIC ACID, OR PLACEBO. METHODS: THIRTY FEMALE PATIENTS WERE CLINICALLY EVALUATED FOR THE EXPRESSION OF DNMT1 AND DNMT3B USING REAL-TIME PCR AND IMMUNOFLUORESCENCE. THESE INITIAL RESULTS WERE COMPARED TO RESULTS AFTER EIGHT WEEKS OF TREATMENT WITH SUNSCREEN IN COMBINATION WITH NIACINAMIDE, RETINOIC ACID, OR PLACEBO. RESULTS: THE RELATIVE EXPRESSION OF DNMT1 WAS SIGNIFICANTLY ELEVATED IN MELASMA COMPARED WITH UNAFFECTED SKIN IN ALL SUBJECTS, INDICATING DNA HYPERMETHYLATION. AFTER TREATMENT, IT WAS DECREASED IN ALL GROUPS: NIACINAMIDE (7 VERSUS 1; P<0.01), RETINOIC ACID (7 VERSUS 2; P<0.05), AND PLACEBO (7 VERSUS 3; P<0.05), WHICH CORRELATES WITH CLINICAL IMPROVEMENT. DNMT3B WAS NOT OVEREXPRESSED IN LESIONAL SKIN BUT REDUCED IN ALL GROUPS. CONCLUSIONS: WE FOUND DNA HYPERMETHYLATION IN MELASMA LESIONS. ENVIRONMENTAL FACTORS SUCH AS SOLAR RADIATION MAY INDUCE CELLULAR CHANGES THAT TRIGGER HYPERPIGMENTATION THROUGH THE ACTIVATION OF PATHWAYS REGULATED BY EPIGENETIC MODIFICATIONS. HOWEVER, LIMITING OR DECREASING DNA METHYLATION THROUGH SUNSCREEN, NIACINAMIDE, AND RETINOIC ACID TREATMENTS THAT PROVIDE PHOTOPROTECTION AND GENETIC TRANSCRIPTION CAN COUNTERACT THIS.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
6 4360  39 MIR-6769B-5P TARGETS CCND-1 TO REGULATE PROLIFERATION IN CADMIUM-TREATED PLACENTAL TROPHOBLASTS: ASSOCIATION WITH THE IMPAIRMENT OF FETAL GROWTH. ENVIRONMENTAL CADMIUM (CD) IS POSITIVELY ASSOCIATED WITH PLACENTAL IMPAIRMENT AND FETAL GROWTH RETARDATION. NEVERTHELESS, ITS POTENTIAL MECHANISMS REMAIN UNCLEAR. MICRORNAS (MIRNAS) ARE KNOWN TO INFLUENCE PLACENTAL DEVELOPMENT AND FETAL GROWTH. THIS WORK WAS AIMED TO DETERMINE WHICH MIRNAS ARE INVOLVED IN CD-IMPAIRED PLACENTAL AND FETAL DEVELOPMENT BASED ON THE MRNA AND MIRNA EXPRESSION PROFILES ANALYSIS. AS A RESULT, GESTATIONAL CD EXPOSURE DECEASED FETAL AND PLACENTAL WEIGHT, AND REDUCED THE PROTEIN LEVEL OF PCNA IN HUMAN AND MOUSE PLACENTAE. FURTHERMORE, THE RESULTS OF MRNA MICROARRAY SHOWED THAT CD-DOWNREGULATED MRNAS WERE PREDICTIVELY CORRELATED WITH SEVERAL BIOLOGICAL PROCESSES, INCLUDING CELL PROLIFERATION, DIFFERENTIATION AND MOTILITY. IN ADDITION, THE RESULTS OF MIRNA MICROARRAY AND QPCR ASSAY DEMONSTRATED THAT CD SIGNIFICANTLY INCREASED THE LEVEL OF MIR-6769B-5P, MIR-146B-5P AND MIR-452-5P. INTEGRATED ANALYSIS OF CD-UPREGULATED MIRNAS PREDICTED TARGET GENES AND CD-DOWNREGULATED MRNAS FOUND THAT OVERLAPPING MRNAS, SUCH AS CCND1, CDK13, RINT1 AND CDC26 WERE ALSO SIGNIFICANTLY ASSOCIATED WITH CELL PROLIFERATION. FURTHER EXPERIMENTS SHOWED THAT MIR-6769B-5P INHIBITOR, BUT NOT MIR-146B-5P AND MIR-452-5P, MARKEDLY REVERSED CD-DOWNREGULATED THE EXPRESSION OF PROLIFERATION-RELATED MRNAS, AND THEREBY RESTORED CD-DECREASED THE PROTEINS LEVEL OF CCND1 AND PCNA IN HUMAN PLACENTAL TROPHOBLASTS. DUAL LUCIFERASE REPORTER ASSAY FURTHER REVEALED THAT MIR-6769B-5P DIRECTLY TARGETS CCND1. FINALLY, THE CASE-CONTROL STUDY DEMONSTRATED THAT INCREASED MIR-6769B-5P LEVEL AND IMPAIRED CELL PROLIFERATION WERE OBSERVED IN SMALL-FOR-GESTATIONAL-AGE HUMAN PLACENTAE. IN CONCLUSION, MIR-6769B-5P TARGETS CCND-1 TO REGULATE PROLIFERATION IN CD-TREATED PLACENTAL TROPHOBLASTS, WHICH IS ASSOCIATED WITH THE IMPAIRMENT OF FETAL GROWTH. OUR FINDINGS IMPLY THAT PLACENTAL MIR-6769B-5P MAY BE USED AS AN EPIGENETIC MARKER FOR ENVIRONMENTAL POLLUTANTS-CAUSED FETAL GROWTH RESTRICTION AND ITS LATE-ONSET CHRONIC DISEASES.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
7 5205  29 PRENATAL STRESS CHANGES THE GLYCOPROTEIN GPM6A GENE EXPRESSION AND INDUCES EPIGENETIC CHANGES IN RAT OFFSPRING BRAIN. PRENATAL STRESS (PS) EXERTS STRONG IMPACT ON FETAL BRAIN DEVELOPMENT AND ON ADULT OFFSPRING BRAIN FUNCTIONS. PREVIOUS WORK DEMONSTRATED THAT CHRONIC STRESS ALTERS THE MRNA EXPRESSION OF GPM6A, A NEURONAL GLYCOPROTEIN INVOLVED IN FILOPODIUM EXTENSION. IN THIS WORK, WE ANALYZED THE EFFECT OF PS ON GPM6A EXPRESSION AND THE EPIGENETIC MECHANISMS INVOLVED. PREGNANT WISTAR RATS RECEIVED RESTRAINT STRESS DURING THE LAST WEEK OF GESTATION. MALE OFFSPRING WERE SACRIFICED ON POSTNATAL DAYS 28 AND 60. HIPPOCAMPUS AND PREFRONTAL CORTEX SAMPLES WERE ANALYZED FOR GENE EXPRESSION (QPCR FOR MRNAS AND MICRORNAS), METHYLATION STATUS (BISULFITE CONVERSION) AND PROTEIN LEVELS. HIPPOCAMPAL NEURONS IN CULTURE WERE USED TO ANALYZE MICRORNA OVEREXPRESSION EFFECTS. PRENATAL STRESS INDUCED CHANGES IN GPM6A LEVELS IN BOTH TISSUES AND AT BOTH AGES ANALYZED, INDICATING A PERSISTENT EFFECT. TWO CPG ISLANDS IN THE GPM6A GENE WERE IDENTIFIED. VARIATIONS IN THE METHYLATION PATTERN AT THREE SPECIFIC CPGS WERE FOUND IN HIPPOCAMPUS, BUT NOT IN PFC SAMPLES FROM PS OFFSPRING. MICRORNAS PREDICTED TO TARGET GPM6A WERE IDENTIFIED IN SILICO. QPCR MEASUREMENTS SHOWED THAT PS MODIFIED THE EXPRESSION OF SEVERAL MICRORNAS IN BOTH TISSUES, BEING MICRORNA-133B THE MOST SIGNIFICANTLY ALTERED. FURTHER STUDIES OVEREXPRESSING THIS MICRORNA IN NEURONAL CULTURES SHOWED A REDUCTION IN GMP6A MRNA AND PROTEIN LEVEL. MOREOVER FILOPODIUM DENSITY WAS ALSO REDUCED, SUGGESTING THAT GPM6A FUNCTION WAS AFFECTED. GESTATIONAL STRESS AFFECTED GPM6A GENE EXPRESSION IN OFFSPRING LIKELY THROUGH CHANGES IN METHYLATION STATUS AND IN POSTTRANSCRIPTIONAL REGULATION BY MICRORNAS. THUS, OUR FINDINGS PROPOSE GPM6A AS A NOVEL TARGET FOR EPIGENETIC REGULATION DURING PRENATAL STRESS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
8 2758  28 EXPRESSION OF HORMONAL CARCINOGENESIS GENES AND RELATED REGULATORY MICRORNAS IN UTERUS AND OVARIES OF DDT-TREATED FEMALE RATS. THE INSECTICIDE DICHLORODIPHENYLTRICHLOROETHANE (DDT) IS A NONMUTAGENIC XENOBIOTIC COMPOUND ABLE TO EXERT ESTROGEN-LIKE EFFECTS RESULTING IN ACTIVATION OF ESTROGEN RECEPTOR-ALPHA (ERALPHA) FOLLOWED BY CHANGED EXPRESSION OF ITS DOWNSTREAM TARGET GENES. IN ADDITION, STUDIES PERFORMED OVER RECENT YEARS SUGGEST THAT DDT MAY ALSO INFLUENCE EXPRESSION OF MICRORNAS. HOWEVER, AN IMPACT OF DDT ON EXPRESSION OF ER, MICRORNAS, AND RELATED TARGET GENES HAS NOT BEEN FULLY ELUCIDATED. HERE, USING REAL-TIME PCR, WE ASSESSED CHANGES IN EXPRESSION OF KEY GENES INVOLVED IN HORMONAL CARCINOGENESIS AS WELL AS POTENTIALLY RELATED REGULATORY ONCOGENIC/TUMOR SUPPRESSOR MICRORNAS AND THEIR TARGET GENES IN THE UTERUS AND OVARIES OF FEMALE WISTAR RATS DURING SINGLE AND CHRONIC MULTIPLE-DOSE DDT EXPOSURE. WE FOUND THAT APPLYING DDT RESULTS IN ALTERED EXPRESSION OF MICRORNAS-221, -222, -205, -126A, AND -429, THEIR TARGET GENES (PTEN, DICER1), AS WELL AS GENES INVOLVED IN HORMONAL CARCINOGENESIS (ESR1, PGR, CCND1, CYP19A1). NOTABLY, CYP19A1 EXPRESSION SEEMS TO BE ALSO REGULATED BY MICRORNAS-221, -222, AND -205. THE DATA SUGGEST THAT EPIGENETIC EFFECTS INDUCED BY DDT AS A POTENTIAL CARCINOGEN MAY BE BASED ON AT LEAST TWO MECHANISMS: (I) ACTIVATION OF ERALPHA FOLLOWED BY ALTERED EXPRESSION OF THE TARGET GENES ENCODING RECEPTOR PGR AND CCND1 AS WELL AS IMPAIRED EXPRESSION OF CYP19A1, AFFECTING, THEREBY, CELL HORMONE BALANCE; AND (II) CHANGED EXPRESSION OF MICRORNAS RESULTING IN IMPAIRED EXPRESSION OF RELATED TARGET GENES INCLUDING REDUCED LEVEL OF CYP19A1 MRNA.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
9 3657  33 INDUCTION AND RECOVERY OF CPG SITE SPECIFIC METHYLATION CHANGES IN HUMAN BRONCHIAL CELLS AFTER LONG-TERM EXPOSURE TO CARBON NANOTUBES AND ASBESTOS. INTRODUCTION: INHALATION OF ASBESTOS INDUCES LUNG CANCER VIA DIFFERENT CELLULAR MECHANISMS. TOGETHER WITH THE INCREASED PRODUCTION OF CARBON NANOTUBES (CNTS) GROWS THE CONCERN ABOUT ADVERSE EFFECTS ON THE LUNGS GIVEN THE SIMILARITIES WITH ASBESTOS. WHILE IT HAS BEEN ESTABLISHED THAT CNT AND ASBESTOS INDUCE EPIGENETIC ALTERATIONS, IT IS CURRENTLY NOT KNOWN WHETHER ALTERATIONS AT EPIGENETIC LEVEL REMAIN STABLE AFTER WITHDRAWAL OF THE EXPOSURE. IDENTIFICATION OF DNA METHYLATION CHANGES AFTER A LOW DOSE OF CNT AND ASBESTOS EXPOSURE AND RECOVERY CAN BE USEFUL TO DETERMINE THE FIBRE/PARTICLE TOXICITY AND ADVERSE OUTCOME. METHODS: HUMAN BRONCHIAL EPITHELIAL CELLS (16HBE) WERE TREATED WITH A LOW AND NON-CYTOTOXIC DOSE (0.25 MICROG/ML) OF MULTI-WALLED CARBON NANOTUBES (MWCNTS-NM400) OR SINGLE-WALLED CARBON NANOTUBES (SWCNTS-SRM2483) AND 0.05 MICROG/ML AMOSITE (BROWN) ASBESTOS FOR THE COURSE OF FOUR WEEKS (SUB-CHRONIC EXPOSURE). AFTER THIS TREATMENT, THE CELLS WERE FURTHER INCUBATED (WITHOUT PARTICLE/FIBRE) FOR TWO WEEKS, ALLOWING RECOVERY FROM THE EXPOSURE (RECOVERY PERIOD). NUCLEAR DEPOSITIONS OF THE CNTS WERE ASSESSED USING FEMTOSECOND PULSED LASER MICROSCOPY IN A LABEL-FREE MANNER. DNA METHYLATION ALTERATIONS WERE ANALYSED USING MICROARRAYS THAT ASSESS MORE THAN 850 THOUSAND CPG SITES IN THE WHOLE GENOME. RESULTS: AT NON-CYTOTOXIC DOSES, CNTS WERE NOTED TO BE INCORPORATED WITH IN THE NUCLEUS AFTER A FOUR WEEKS PERIOD. EXPOSURE TO MWCNTS INDUCED A SINGLE HYPOMETHYLATION AT A CPG SITE AND GENE PROMOTER REGION. NO CHANGE IN DNA METHYLATION WAS OBSERVED AFTER THE RECOVERY PERIOD FOR MWCNTS. EXPOSURE TO SWCNTS OR AMOSITE INDUCED HYPERMETHYLATION AT CPG SITES AFTER SUB-CHRONIC EXPOSURE WHICH MAY INVOLVE IN 'TRANSCRIPTION FACTOR ACTIVITY' AND 'SEQUENCE-SPECIFIC DNA BINDING' GENE ONTOLOGIES. AFTER THE RECOVERY PERIOD, HYPERMETHYLATION AND HYPOMETHYLATION WERE NOTED FOR BOTH SWCNTS AND AMOSITE. HIPPOCALCINLIKE 1 (HPCAL1), PROTEASE SERINE 3 (PRSS3), KALLIKREIN-RELATED PEPTIDASE 3 (KLK3), KRUPPEL LIKE FACTOR 3 (KLF3) GENES WERE HYPERMETHYLATED AT DIFFERENT TIME POINTS IN EITHER SWCNT-EXPOSED OR AMOSITE-EXPOSED CELLS. CONCLUSION: THESE RESULTS SUGGEST THAT THE SPECIFIC SWCNT (SRM2483) AND AMOSITE FIBRES STUDIED INDUCE HYPO- OR HYPERMETHYLATION ON CPG SITES IN DNA AFTER VERY LOW-DOSE EXPOSURE AND RECOVERY PERIOD. THIS EFFECT WAS NOT SEEN FOR THE STUDIED MWCNT (NM400).	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
10 6835  27 [INFLUENCE OF AGE OF PATIENTS WITH BRONCHOPULMONARY PATHOLOGY ON LOW-MOLECULAR DNA CONCENTRATION IN BLOOD PLASMA.]. THE AIM OF THE WORK WAS TO DETERMINE THE CONCENTRATION OF LOW-MOLECULAR-WEIGHT PLASMA DNA (LMDNA) IN PATIENTS WITH COPD AND CHRONIC NON-OBSTRUCTIVE BRONCHITIS (CNONB) OF TWO AGE GROUPS - 34-59 AND 60-80 YEARS. THE LEVELS OF LMDNA IN HEALTHY DONORS, PATIENTS WITH CNONB, HEALTHY RELATIVES OF PATIENTS WITH COPD DID NOT DIFFER, WHILE THE CONCENTRATION OF LMDNA IN PATIENTS WITH COPD WAS SIGNIFICANTLY LOWER. IN COPD PATIENTS AGED 34-59 YEARS, THE LEVEL OF LMDNA WAS REDUCED BY MORE THAN 7 TIMES, AND IN COPD PATIENTS WHO SURVIVED TO 60-80 YEARS, IT WAS 3 TIMES LOWER COMPARED TO THE VALUE OF THIS BIOCHEMICAL INDICATOR IN HEALTHY DONORS OF THE SAME AGE. THE REDUCTION OF LMDNA REFLECTED A REDUCED SYSTEMIC APOPTOTIC ACTIVITY IN THE BODY OF PATIENTS WITH COPD. A SIGNIFICANT DIFFERENCE IN THE CONCENTRATION OF LMDNA IN PATIENTS WITH COPD AND CNONB IN REMISSION CAN BE USED FOR DIFFERENTIAL DIAGNOSIS OF THE DEVELOPMENT OF THESE PATHOLOGICAL PROCESSES. AN INCREASE IN THE LOW LEVEL OF LMDNA IN COPD PATIENTS DURING AGING MAY INDICATE THE INVOLVEMENT OF EPIGENETIC MECHANISMS IN LIFE EXTENSION.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
11 3907  30 LEUCOCYTIC DNA METHYLATION OF INTERLEUKIN-6 PROMOTER REDUCTION IN PRE-HYPERTENSIVE YOUNG ADULTS. BACKGROUND: PRE-HYPERTENSION IS ASSOCIATED WITH INCREASED RISK OF CARDIOVASCULAR DISEASE. CHRONIC INFLAMMATION PLAYS AN IMPORTANT ROLE IN THE PATHOPHYSIOLOGY OF ESSENTIAL HYPERTENSION, WITH EPIGENETIC DYSREGULATION INVOLVEMENT. NEVERTHELESS, THE ROLE OF DNA METHYLATION IN PREHYPERTENSIVE STATE IS UNKNOWN. THE AIM OF THIS STUDY WAS TO INVESTIGATE THE ASSOCIATION BETWEEN DNA METHYLATION LEVEL OF INTERLEUKIN-6 (IL-6) PROMOTER IN PRE-HYPERTENSIVE (PREHT) AND NORMOTENSIVE (NT) YOUNG ADULTS. METHODS: A TOTAL OF 80 NT AND 80 PREHT HEALTHY SUBJECTS AGED BETWEEN 18-45 YEARS WERE RECRUITED IN KUANTAN, PAHANG, MALAYSIA USING AN OBSERVATIONAL CROSS-SECTIONAL STUDY APPROACH. DNA METHYLATION LEVEL OF IL-6 PROMOTER IN PERIPHERAL LEUKOCYTES WERE MEASURED USING BISULPHITE CONVERSION AND METHYLIGHT ASSAY. RESULTS: THERE WAS NO SIGNIFICANT DIFFERENCE IN AGE BETWEEN NT AND PREHT (P = 0.655). THE MEAN BLOOD PRESSURE WAS 110(8)/73(5) MMHG IN NT AND 125(7)/82(5) MMHG IN PREHT SUBJECTS. THE IL-6 PROMOTER METHYLATION LEVEL WAS SIGNIFICANTLY LOWER IN PREHT COMPARED TO NT SUBJECTS (P < 0.001). CONCLUSION: THE CURRENT STUDY DEMONSTRATES THAT HYPOMETHYLATION OF IL-6 PROMOTER WAS ASSOCIATED WITH PRE-HYPERTENSION IN YOUNG ADULTS. THUS, IL-6 METHYLATION COULD BE USED AS AN EARLY INDICATOR FOR PREDICTING HYPERTENSION AND RELATED RISK OF CARDIOVASCULAR DISEASES IN PREHYPERTENSIVE SUBJECTS. GENE EXPRESSION AND LONGITUDINAL STUDIES ARE WARRANTED TO EXAMINE THE METHYLATION EFFECT ON IL-6 EXPRESSION OVER TIME.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
12 1187  29 COPD GWAS VARIANT AT 19Q13.2 IN RELATION WITH DNA METHYLATION AND GENE EXPRESSION. CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) IS AMONG THE MAJOR HEALTH BURDENS IN ADULTS. WHILE CIGARETTE SMOKING IS THE LEADING RISK FACTOR, A GROWING NUMBER OF GENETIC VARIATIONS HAVE BEEN DISCOVERED TO INFLUENCE DISEASE SUSCEPTIBILITY. EPIGENETIC MODIFICATIONS MAY MEDIATE THE RESPONSE OF THE GENOME TO SMOKING AND REGULATE GENE EXPRESSION. CHROMOSOME 19Q13.2 REGION IS ASSOCIATED WITH BOTH SMOKING AND COPD, YET ITS FUNCTIONAL ROLE IS UNCLEAR. OUR STUDY AIMED TO DETERMINE WHETHER RS7937 (RAB4B, EGLN2), A TOP GENETIC VARIANT IN 19Q13.2 REGION IDENTIFIED IN GENOME-WIDE ASSOCIATION STUDIES OF COPD, IS ASSOCIATED WITH DIFFERENTIAL DNA METHYLATION IN BLOOD (N = 1490) AND GENE EXPRESSION IN BLOOD (N = 721) AND LUNGS (N = 1087). WE COMBINED GENETIC AND EPIGENETIC DATA FROM THE ROTTERDAM STUDY (RS) TO PERFORM THE EPIGENOME-WIDE ASSOCIATION ANALYSIS OF RS7937. FURTHER, WE USED GENETIC AND TRANSCRIPTOMIC DATA FROM BLOOD (RS) AND FROM LUNG TISSUE (LUNG EXPRESSION QUANTITATIVE TRAIT LOCI MAPPING STUDY), TO PERFORM THE TRANSCRIPTOME-WIDE ASSOCIATION STUDY OF RS7937. RS7937 WAS SIGNIFICANTLY (FDR < 0.05) AND CONSISTENTLY ASSOCIATED WITH DIFFERENTIAL DNA METHYLATION IN BLOOD AT 4 CPG SITES IN CIS, INDEPENDENT OF SMOKING. ONE METHYLATION SITE (CG11298343-EGLN2) WAS ALSO ASSOCIATED WITH COPD (P = 0.001). ADDITIONALLY, RS7937 WAS ASSOCIATED WITH GENE EXPRESSION LEVELS IN BLOOD IN CIS (EGLN2), 42% MEDIATED THROUGH CG11298343, AND IN LUNG TISSUE, IN CIS AND TRANS (NUMBL, EGLN2, DNMT3A, LOC101929709 AND PAK2). OUR RESULTS SUGGEST THAT CHANGES OF DNA METHYLATION AND GENE EXPRESSION MAY BE INTERMEDIATE STEPS BETWEEN GENETIC VARIANTS AND COPD, BUT FURTHER CAUSAL STUDIES IN LUNG TISSUE SHOULD CONFIRM THIS HYPOTHESIS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
13 1067  31 CLINICAL UTILITY OF PDSS2 EXPRESSION TO STRATIFY PATIENTS AT RISK FOR RECURRENCE OF HEPATOCELLULAR CARCINOMA. IDENTIFICATION OF NOVEL GENETIC AND EPIGENETIC ALTERATIONS IS REQUIRED FOR OPTIMAL STRATIFICATION OF PATIENTS WITH HEPATOCELLULAR CARCINOMA (HCC) AT RISK FOR RECURRENCE AND ADVERSE PROGNOSIS. COENZYME Q10 (COQ10), WHICH MEDIATES APOPTOSIS, IS SYNTHESIZED BY PRENYL DIPHOSPHATE SYNTHASE SUBUNIT 2 (PDSS2). IN THE PRESENT STUDY WE EVALUATED THE CLINICAL SIGNIFICANCE AND REGULATORY MECHANISMS OF PDSS2 EXPRESSION IN HCC. PDSS2 EXPRESSION LEVELS AND THOSE OF GENES ENCODING POTENTIALLY INTERACTING PROTEINS AS WELL AS THE METHYLATION STATUS OF THE PDSS2 PROMOTER REGION WERE ANALYZED IN HCC CELL LINES. PDSS2 MRNA LEVELS IN 151 PAIRS OF RESECTED SPECIMENS WERE DETERMINED TO EVALUATE THE ASSOCIATION OF PDSS2 EXPRESSION AND CLINICOPATHOLOGICAL FACTORS. THE EXPRESSION AND DISTRIBUTION OF PDSS2 WERE DETERMINED USING IMMUNOHISTOCHEMISTRY. PDSS2 MRNA EXPRESSION WAS DECREASED IN SIX OF NINE HCC CELL LINES AND SIGNIFICANTLY CORRELATED WITH THOSE OF HEPATOCYTE NUCLEAR FACTOR 4ALPHA. PDSS2 TRANSCRIPTION IN HCC CELLS WITH DECREASED PDSS2 EXPRESSION ACCOMPANYING HYPERMETHYLATION WAS REACTIVATED AFTER TREATING THESE CELLS WITH A METHYLATION INHIBITOR. MEAN EXPRESSION LEVELS OF PDSS2 MRNA RELATIVE TO THAT OF UNINVOLVED LIVER DIMINISHED GRADUALLY IN THE ORDER OF CHRONIC HEPATITIS TO CIRRHOSIS, AND EACH WAS SIGNIFICANTLY HIGHER THAN THOSE OF HCCS. PDSS2 AND PDSS2 MRNA LEVELS WERE CONSISTENT. DECREASED PDSS2 MRNA LEVELS WERE DETECTED IN HCC TISSUES OF 56 PATIENTS, CORRELATED WITH SHORTER DISEASE-SPECIFIC SURVIVAL, AND WAS IDENTIFIED AS AN INDEPENDENT PROGNOSTIC FACTOR. PDSS2 IS A PUTATIVE TUMOR SUPPRESSOR, AND PROMOTER HYPERMETHYLATION IS A KEY REGULATORY MECHANISM IN HCC. DECREASED LEVELS OF PDSS2 MRNA EXPRESSION MAY REPRESENT A NOVEL BIOMARKER OF HCC.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
14 6465  19 TISSUE REMODELING IN ADULT VERNAL KERATOCONJUNCTIVITIS. OUR AIM IS TO DESCRIBE LOCAL TISSUE REMODELING IN A COHORT OF ADULT VKC PATIENTS. MALE PATIENTS DIAGNOSED WITH ACTIVE VKC WERE ENROLLED IN AN OPEN PILOT STUDY INTO TWO GROUPS ACCORDING DISEASE ONSET: CHILDHOOD CLASSIC VKC AND ADULT VKC. VISUAL ACUITY AND OCULAR SURFACE CLINICAL EXAMINATION FOCUSING ON CHRONIC INFLAMMATORY SEQUELAE AND IMPRESSION CYTOLOGY WERE PERFORMED IN ALL ENROLLED SUBJECTS. CONJUNCTIVAL IMPRINTS WERE PROCESSED FOR MOLECULAR, BIOCHEMICAL AND IMMUNOFLUORESCENT ANALYSIS FOR TISSUE REMODELING (TGFBETA1,2,3 AND ALPHASMA) AND EPIGENETIC (DNMT3A, KEAP1; NRF2) MARKERS AS WELL AS ANDROGEN RECEPTORS WERE INVESTIGATED AND COMPARED BETWEEN GROUPS. CLINICAL ASSESSMENT SHOWED INCREASED CONJUNCTIVAL SCARRING IN ADULT VKC COMPARED TO CLASSIC VKC. IMMUNOREACTIVITY FOR ALPHASMA AND EXPRESSION OF TGFBETA WERE HIGHER IN ADULT VKC GROUP. SIGNIFICANTLY HIGHER LEVELS OF TGFBETA3 (3.44 +/- 1.66; P < 0.05) WERE DETECTED IN ADULT VKC COMPARED TO CHILDHOOD VKC, ASSOCIATED WITH AN INCREASING TREND OF TGFBETA1 (1.58 +/- 0.25) AND TGFBETA2 (1.65 +/- 0.20) ISOFORMS LEVELS. MOLECULAR ANALYSIS SHOWED A RELATIVE INCREASE IN TISSUE REMODELING/FIBROGENIC TRANSCRIPTS (TGFBETA ISOFORMS AND ALPHASMA) ASSOCIATED TO A SIGNIFICANT INCREASE OF SELECTIVE EPIGENETIC TARGETS (DNMT3, NRF2 AND KEAP1) IN ADULT VKC PHENOTYPE. INCREASED LOCAL CONJUNCTIVAL ANDROGEN RECEPTORS WAS DETECTED IN PATIENTS WITH ADULT VARIANTS COMPARED TO CLASSIC CHILDHOOD VKC AND HEALTHY SUBJECTS. FINALLY, A DIRECT CORRELATION BETWEEN TGFBETA AND ANDROGEN RECEPTOR EXPRESSION WAS ALSO DETECTED. A PRO-FIBROTIC CLINICAL AND BIOMOLECULAR TRAIT WAS UNVEILED IN ADULT VARIANT OF VKC, WHICH CAUSES OCULAR SURFACE DISEASE AND VISUAL IMPAIRMENT.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
15  921  30 CHRONIC HYPOXIA-INDUCED CIRBP HYPERMETHYLATION ATTENUATES HYPOTHERMIC CARDIOPROTECTION VIA DOWN-REGULATION OF UBIQUINONE BIOSYNTHESIS. THERAPEUTIC HYPOTHERMIA IS COMMONLY USED DURING CARDIOPULMONARY BYPASS (CPB) TO PROTECT THE HEART AGAINST MYOCARDIAL INJURY IN CARDIAC SURGERY. PATIENTS WHO SUFFER FROM CHRONIC HYPOXIA (CH), SUCH AS THOSE WITH CERTAIN HEART OR LUNG CONDITIONS, ARE AT HIGH RISK OF SEVERE MYOCARDIAL INJURY AFTER CARDIAC SURGERY, BUT THE UNDERLYING MECHANISMS ARE UNKNOWN. THIS STUDY TESTED WHETHER CH ATTENUATES HYPOTHERMIC CARDIOPROTECTION DURING CPB. USING A RAT MODEL OF CPB, WE FOUND THAT HYPOTHERMIC CARDIOPROTECTION WAS IMPAIRED IN CH RATS BUT WAS PRESERVED IN NORMOXIC RATS. CARDIAC PROTEOMES SHOWED THAT COLD-INDUCIBLE RNA BINDING PROTEIN (CIRBP) WAS SIGNIFICANTLY (P = 0.03) DECREASED IN CH RATS DURING CPB. METHYLATION ANALYSIS OF NEONATAL RAT CARDIOMYOCYTES UNDER CH AND MYOCARDIUM SPECIMENS FROM PATIENTS WITH CH SHOWED THAT CH INDUCED HYPERMETHYLATION OF THE CIRBP PROMOTER REGION, RESULTING IN ITS DEPRESSION AND FAILURE TO RESPOND TO COLD STRESS. CIRBP-KNOCKOUT RATS SHOWED ATTENUATED HYPOTHERMIC CARDIOPROTECTION, WHEREAS CIRBP-TRANSGENIC RATS SHOWED AN ENHANCED RESPONSE. PROTEOMICS ANALYSIS REVEALED THAT THE CARDIAC UBIQUINONE BIOSYNTHESIS PATHWAY WAS DOWN-REGULATED DURING CPB IN CIRBP-KNOCKOUT RATS, RESULTING IN A SIGNIFICANTLY (P = 0.01) DECREASED CONCENTRATION OF UBIQUINONE (COQ(10)). CONSEQUENTLY, CARDIAC OXIDATIVE STRESS WAS AGGRAVATED AND ADENOSINE 5'-TRIPHOSPHATE PRODUCTION WAS IMPAIRED, LEADING TO INCREASED MYOCARDIAL INJURY DURING CPB. COQ(10)-SUPPLEMENTED CARDIOPLEGIC SOLUTION IMPROVED CARDIOPROTECTION IN RATS EXPOSED TO CH, BUT ITS EFFECT WAS LIMITED IN NORMOXIC RATS. OUR STUDY SUGGESTS THAT AN INDIVIDUALIZED CARDIOPROTECTION STRATEGY SHOULD BE USED TO FULLY COMPENSATE FOR THE CONSEQUENCES OF EPIGENETIC MODIFICATION OF CIRBP IN PATIENTS WITH CH WHO REQUIRE THERAPEUTIC HYPOTHERMIA.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
16 2761  31 EXPRESSION OF TESTIS-SPECIFIC GENES, TEX101 AND ODF4, IN CHRONIC MYELOID LEUKEMIA AND EVALUATION OF TEX101 IMMUNOGENICITY. BACKGROUND AND OBJECTIVES: CANCER-TESTIS (CT) ANTIGENS ARE A GROUP OF ANTIGENS WITH A RESTRICTED EXPRESSION IN NORMAL TISSUES, EXCEPT TESTIS, AND THEY HAVE ABERRANT EXPRESSION IN DIFFERENT TUMORS. THIS PATTERN OF EXPRESSION HAS MADE THEM PROMISING TARGETS FOR IMMUNOTHERAPY AND CANCER DETECTION. OUR AIM WAS TO FIND NEW MEMBERS OF THIS GROUP THAT MIGHT BE USEFUL AS MARKERS IN THE DETECTION OF CANCER AND IMMUNOTHERAPY. DESIGN AND SETTING: A DESCRIPTIVE STUDY CONDUCTED IN REFERRAL CENTERS OF TEHRAN UNIVERSITY OF MEDICAL SCIENCE FROM JANUARY 2008 TO JANUARY 2009. PATIENTS AND METHODS: WE ANALYZED THE EXPRESSION OF TWO TESTIS-SPECIFIC GENES NAMED ODF4 (OUTER DENSE FIBER OF SPERM TAILS 4) AND TEX101 (TESTIS EXPRESSED 101) IN 20 CHRONIC MYELOID LEUKEMIA (CML) AND 20 NORMAL SAMPLES BY REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION AND SEQUENCING. IMMUNOGENICITY OF TEX101 WAS EVALUATED BY MEANS OF ENZYME-LINKED IMMUNOSORBENT ASSAY. RESULTS: THESE TWO GENES WERE EXPRESSED IN 30% OF CML PATIENTS BUT NOT IN ANY OF THE HEALTHY DONORS. HUMORAL RESPONSE AGAINST TEX101 WAS NOT DETECTED IN ANY SAMPLES. CONCLUSIONS: TEX101 AND ODF4 ARE CT GENES USEFUL FOR DETECTION OF CML. UNLIKE MANY CT GENES, OVEREXPRESSION OF TEX101 WAS NOT SHOWN TO INDUCE IMMUNOLOGIC RESPONSES IN THESE SAMPLES. ACCORDING TO THE PREVIOUS STUDIES, OVEREXPRESSION OF TEX101 LEADS TO SUPPRESSION OF CANCER INVASION AND METASTASIS; THUS, THE INDUCTION OF THE EXPRESSION OF TEX101 IN CANCER BY EPIGENETIC MECHANISMS MAY BE A TREATMENT STRATEGY.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
17  441  34 ANTISENSE-INDUCED DOWNREGULATION OF MAJOR CIRCADIAN GENES MODULATES THE EXPRESSION OF HISTONE DEACETYLASE-2 (HDAC-2) AND CREB-BINDING PROTEIN (CBP) IN THE MEDIAL SHELL REGION OF NUCLEUS ACCUMBENS OF MICE EXPOSED TO CHRONIC EXCESSIVE ALCOHOL CONSUMPTION. CIRCADIAN GENES IN THE MEDIAL ACCUMBAL SHELL (MNACSH) REGION REGULATE BINGE ALCOHOL CONSUMPTION. HERE, WE INVESTIGATED IF ANTISENSE-INDUCED KNOCKDOWN OF MAJOR CIRCADIAN GENES (PER1, PER2, AND NPAS2) IN THE MNACSH OF MICE EXPOSED TO INTERMITTENT ACCESS TWO-BOTTLE CHOICE (IA2BC) PARADIGM MODULATES THE EXPRESSION OF HISTONE DEACETYLASE-2 (HDAC-2) AND CREB-BINDING PROTEIN (CBP), KEY EPIGENETIC MODIFIERS ASSOCIATED WITH WITHDRAWAL-ASSOCIATED BEHAVIORS SUCH AS ANXIETY. ADULT MALE C57BL/6J MICE (N = 28), SURGICALLY IMPLANTED WITH BILATERAL GUIDE CANNULAS ABOVE THE MNACSH, WERE CHRONICALLY (4 WEEKS) EXPOSED TO ALCOHOL (20% V/V) OR SACCHARIN (0.03%) VIA IA2BC PARADIGM. IN THE FOURTH WEEK, A MIXTURE OF ANTISENSE (AS-ODNS; N = 14/GROUP) OR NONSENSE (NS-ODNS; N = 14/GROUP) OLIGODEOXYNUCLEOTIDES AGAINST CIRCADIAN GENES WERE BILATERALLY INFUSED INTO THE MNACSH. SUBSEQUENTLY, ALCOHOL/SACCHARIN CONSUMPTION AND PREFERENCE WERE MEASURED FOLLOWED BY EUTHANIZATION OF ANIMALS AND VERIFICATION OF MICROINJECTION SITES BY VISUAL INSPECTION AND THE EXPRESSION OF HDAC-2 AND CBP BY USING RT-PCR ALONG WITH THE VERIFICATION OF ANTISENSE-INDUCED DOWNREGULATION OF CIRCADIAN GENES IN THE MNACSH. AS COMPARED WITH NS-ODNS, AS-ODNS INFUSION SIGNIFICANTLY ATTENUATED THE ALCOHOL-INDUCED INCREASE IN HDAC-2 AND REDUCTION IN CBP EXPRESSION IN THE MNACSH ALONG WITH A SIGNIFICANT REDUCTION IN ALCOHOL CONSUMPTION AND PREFERENCE. NO SIGNIFICANT EFFECT WAS OBSERVED ON EITHER SACCHARIN CONSUMPTION OR PREFERENCE. OUR RESULTS SUGGEST THAT CIRCADIAN GENES IN THE MNACSH MAY HAVE A CAUSAL TO PLAY IN MEDIATING EPIGENETIC CHANGES OBSERVED AFTER CHRONIC ALCOHOL CONSUMPTION.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
18 6636  40 UNRAVELING A NEW PLAYER IN MULTIPLE SCLEROSIS PATHOGENESIS: THE RNA-BINDING PROTEIN HUR. BACKGROUND: ELAV-LIKE PROTEINS ARE A SMALL FAMILY OF RNA-BINDING PROTEINS THAT ARE FUNDAMENTAL PLAYERS IN POST-TRANSCRIPTIONAL MECHANISMS AND ARE INVOLVED IN THE PATHOGENESIS OF NEUROLOGIC AND PSYCHIATRIC DISORDERS. HUR, THE UBIQUITOUSLY EXPRESSED MEMBER OF THE FAMILY, IS ALSO IMPLICATED IN SUSTAINING INFLAMMATION AND INFLAMMATORY DISEASES, SUPPORTING THE PRODUCTION OF PRO-INFLAMMATORY CYTOKINES. INFLAMMATION PLAYS A CENTRAL ROLE IN MULTIPLE SCLEROSIS (MS), WHICH REPRESENTS THE MOST COMMON CAUSE OF PERMANENT PHYSICAL DISABILITY IN YOUNG ADULTS. MS IS A CHRONIC AUTOIMMUNE DISEASE AFFECTING THE CENTRAL NERVOUS SYSTEM, WITH A COMPLEX AETIOLOGY INVOLVING GENETIC, ENVIRONMENTAL AND EPIGENETIC FACTORS. NO DATA ARE AVAILABLE ON THE POTENTIAL ENTANGLEMENT OF HUR IN MS PATHOGENESIS IN PATIENTS. IN THE PRESENT WORK, WE AIMED AT EXPLORING HUR PROTEIN LEVELS IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) FROM MS PATIENTS, COMPARED TO HEALTHY CONTROLS. TO FURTHER ELUCIDATE THE POSSIBLE INVOLVEMENT OF HUR IN MS, WE ALSO INVESTIGATED THE RELATIONSHIP BETWEEN THIS SPECIFIC RNA-BINDING PROTEIN AND HSP70-2 PROTEIN, ALSO CONSIDERING THE HSP70-2 RS1061581 POLYMORPHISM, GIVEN THAT HSP70-2 MRNA HAS BEEN REPORTED AS A HUR TARGET AND THIS SPECIFIC POLYMORPHISM TO BE ASSOCIATED WITH MS RISK. METHODS: ALLELES AND GENOTYPES FOR HSP70-2 RS1061581 POLYMORPHISM WERE ASSESSED, BY USING A POLYMERASE CHAIN REACTION-RESTRICTION FRAGMENT LENGTH POLYMORPHISM, FOLLOWED BY DIGESTION WITH RESTRICTION ENZYME, IN MS PATIENTS AND HEALTHY CONTROLS. PBMCS FROM A SUBGROUP OF PATIENTS AND CONTROLS WERE USED TO EVALUATE HUR AND HSP70-2 PROTEIN CONTENT BY WESTERN BLOT. RESULTS: PBMCS FROM 52 MS PATIENTS HAD A LOWER HUR AND HIGHER HSP70-2 PROTEIN CONTENT COMPARED TO 43 HEALTHY CONTROLS. AN INCREASE OF 100 UNITS OF HUR SIGNIFICANTLY DECREASED THE RISK OF DEVELOPING MS BY 9.8% (OR: 0.902, 95% CI: 0.83-0.98), CONTROLLING FOR HSP70-2 PROTEIN EXPRESSION, HSP70-2 RS1061581 GENOTYPE, AGE AND SEX. MOREOVER, HOLDING HUR LEVELS, AN INCREASE OF 100 UNITS OF HSP70-2 PROTEIN SIGNIFICANTLY INCREASED THE MS RISK BY 18.1% (OR: 1.181, 95% CI: 1.03-1.36) AND THE GENETIC SUSCEPTIBILITY OF DEVELOPING MS FOR HSP70-2 RS1061581 GG CARRIERS IS CONFIRMED. OF INTEREST, MS PATIENTS WITH A MODERATE TO SEVERE FORM OF MS (MSSS >/= 3) SHOWED A TREND TOWARDS A REDUCTION OF HUR PROTEIN LEVELS COMPARED TO PATIENTS WITH MILD DISEASE SEVERITY (MSSS < 3). CONCLUSIONS: HUR PROTEIN LEVELS ARE REDUCED IN MS PATIENTS COMPARED TO HEALTHY SUBJECTS, AND THE PROTEIN AMOUNT MAY CONTINUE TO DECLINE WITH DISEASE PROGRESSION, SUGGESTING A PUTATIVE ROLE OF THIS RNA-BINDING PROTEIN. MOREOVER, OUR RESULTS SUGGEST THAT MS PATHOLOGY MAY HAVE DISRUPTED THE LINK BETWEEN HUR AND ITS TARGET TRANSCRIPT HSP70-2. IT WILL BE IMPORTANT TO FURTHER EXPLORE THE EXACT ROLE OF HUR IN MS, CONSIDERING THE COMPLEX INTERPLAY WITH OTHER RNA-BINDING FACTORS AND TARGET MRNAS.	2020	

19 5914  25 TARGETED-BISULFITE SEQUENCE ANALYSIS OF THE METHYLATION OF CPG ISLANDS IN GENES ENCODING PNPLA3, SAMM50, AND PARVB OF PATIENTS WITH NON-ALCOHOLIC FATTY LIVER DISEASE. BACKGROUND & AIMS: THE PATHOGENESIS OF NON-ALCOHOLIC FATTY LIVER DISEASE (NAFLD) IS AFFECTED BY EPIGENETIC FACTORS AS WELL AS BY GENETIC VARIATION. METHODS: WE PERFORMED TARGETED-BISULFITE SEQUENCING TO DETERMINE THE LEVELS OF DNA METHYLATION OF 4 CPG ISLANDS (CPG99, CPG71, CPG26, AND CPG101) IN THE REGULATORY REGIONS OF PNPLA3, SAMM50, PARVB VARIANT 1, AND PARVB VARIANT 2, RESPECTIVELY. WE COMPARED THE LEVELS OF METHYLATION OF DNA IN THE LIVERS OF THE FIRST AND SECOND SETS OF PATIENTS WITH MILD (FIBROSIS STAGES 0 AND 1) OR ADVANCED (FIBROSIS STAGES 2 TO 4) NAFLD AND IN THOSE OF PATIENTS WITH MILD (F0 TO F2) OR ADVANCED (F3 AND F4) CHRONIC HEPATITIS C INFECTION. THE HEPATIC MRNA LEVELS OF PNPLA3, SAMM50, AND PARVB WERE MEASURED USING QPCR. RESULTS: CPG26, WHICH RESIDES IN THE REGULATORY REGION OF PARVB VARIANT 1, WAS MARKEDLY HYPOMETHYLATED IN THE LIVERS OF PATIENTS WITH ADVANCED NAFLD. CONVERSELY, CPG99 IN THE REGULATORY REGION OF PNPLA3 WAS SUBSTANTIALLY HYPERMETHYLATED IN THESE PATIENTS. THESE DIFFERENCES IN DNA METHYLATION WERE REPLICATED IN A SECOND SET OF PATIENTS WITH NAFLD OR CHRONIC HEPATITIS C. PNPLA3 MRNA LEVELS IN THE LIVER OF THE SAME SECTION OF A BIOPSY SPECIMEN USED FOR GENOMIC DNA PREPARATION WERE LOWER IN PATIENTS WITH ADVANCED NAFLD COMPARED WITH THOSE WITH MILD NAFLD AND CORRELATED INVERSELY WITH CPG99 METHYLATION IN LIVER DNA. MOREOVER, THE LEVELS OF CPG99 METHYLATION AND PNPLA3 MRNA WERE AFFECTED BY THE RS738409 GENOTYPE. CONCLUSIONS: HYPOMETHYLATION OF CPG26 AND HYPERMETHYLATION OF CPG99 MAY CONTRIBUTE TO THE SEVERITY OF FIBROSIS IN PATIENTS WITH NAFLD OR CHRONIC HEPATITIS C INFECTION.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
20 2946  29 GENETIC AND EPIGENETIC CHANGES IN THE EUTOPIC ENDOMETRIUM OF WOMEN WITH ENDOMETRIOSIS: ASSOCIATION WITH DECREASED ENDOMETRIAL ALPHAVBETA3 INTEGRIN EXPRESSION. ABOUT 40% OF WOMEN WITH INFERTILITY AND 70% OF WOMEN WITH PELVIC PAIN SUFFER FROM ENDOMETRIOSIS. THE PREGNANCY RATE IN WOMEN UNDERGOING IVF WITH LOW ENDOMETRIAL INTEGRIN ALPHAVBETA3 (LEI) EXPRESSION IS SIGNIFICANTLY LOWER COMPARED TO THE WOMEN WITH HIGH ENDOMETRIAL INTEGRIN ALPHAVBETA3 (HEI). MID-SECRETORY EUTOPIC ENDOMETRIAL BIOPSIES WERE OBTAINED FROM HEALTHY CONTROLS (C; N=3), AND WOMEN WITH HEI (N=4) AND LEI (N=4) AND ENDOMETRIOSIS. CHANGES IN GENE EXPRESSION WERE ASSESSED USING HUMAN GENE ARRAYS AND DNA METHYLATION DATA WERE DERIVED USING 385 K TWO-ARRAY PROMOTER ARRAYS. TRANSCRIPTIONAL ANALYSIS REVEALED THAT LEI AND C GROUPS CLUSTERED SEPARATELY WITH 396 DIFFERENTIALLY EXPRESSED GENES (DEGS) (P<0.01: 275 UP AND 121 DOWN) DEMONSTRATING THAT TRANSCRIPTIONAL AND EPIGENETIC CHANGES ARE DISTINCT IN THE LEI EUTOPIC ENDOMETRIUM COMPARED TO THE C AND HEI GROUP. IN CONTRAST, HEI VS C AND HEI VS LEI COMPARISONS ONLY IDENTIFIED 83 AND 45 DEGS, RESPECTIVELY. THE METHYLATION PROMOTER ARRAY IDENTIFIED 1304 DIFFERENTIALLY METHYLATED REGIONS IN THE LEI VS C COMPARISON. THE OVERLAP OF GENE AND METHYLATION ARRAY DATA IDENTIFIED 14 EPIGENETICALLY DYSREGULATED GENES AND QUANTITATIVE RT-PCR ANALYSIS VALIDATED THE TRANSCRIPTOMIC FINDINGS. THE ANALYSIS ALSO REVEALED THAT ARYL HYDROCARBON RECEPTOR (AHR) WAS HYPOMETHYLATED AND SIGNIFICANTLY OVEREXPRESSED IN LEI SAMPLES COMPARED TO C. FURTHER ANALYSIS VALIDATED THAT AHR TRANSCRIPT AND PROTEIN EXPRESSION ARE SIGNIFICANTLY (P<0.05) INCREASED IN LEI WOMEN COMPARED TO C. THE INCREASE IN AHR, TOGETHER WITH THE ALTERED METHYLATION STATUS OF THE 14 ADDITIONAL GENES, MAY PROVIDE A DIAGNOSTIC TOOL TO IDENTIFY THE SUBSET OF WOMEN WHO HAVE ENDOMETRIOSIS-ASSOCIATED INFERTILITY.	2021