1 6768 117 ZOLEDRONIC ACID EPIGENETICALLY ALLEVIATES HIGH-GLUCOSE-SUPPRESSED OSTEOGENIC DIFFERENTIATION OF MC3T3-E1 CELLS. OBJECTIVE: DUE TO THE IMPACT OF EXCESSIVE GLUCOSE ON OSTEOGENIC DIFFERENTIATION, DIABETIC OSTEOPATHY FREQUENTLY RESULTS IN SKELETAL FRAGILITY, OSTEOPOROSIS, AND BONE PAIN. ZOLEDRONIC ACID, A BISPHOSPHONATE (BP) THAT EFFECTIVELY INHIBITS OSTEOCLASTIC BONE RESORPTION IS GIVEN YEARLY TO IMPROVE BONE MINERAL DENSITY (BMD) IN PATIENTS WITH OSTEOPOROSIS. HOWEVER, THE DETAILED MOLECULAR MECHANISMS REMAINED UNCLEAR. THIS STUDY INVESTIGATES THE POSSIBLE PATHWAYS BY WHICH ZOLEDRONIC ACID REGULATES OSTEOGENESIS WHEN BLOOD GLUCOSE LEVELS ARE HIGH. MATERIALS AND METHODS: MC3T3-E1 CELLS WERE TREATED WITH ONE MM ZOLEDRONIC ACID OR NOT IN A STANDARD OR HIGH GLUCOSE CULTURE MEDIUM. A QUANTITATIVE POLYMERASE CHAIN REACTION (QPCR) ASSAY WAS UTILIZED TO ASSESS THE EXPRESSION OF THE TARGET CANDIDATE GENES, INCLUDING RUNX2, MALAT1, MIR-133, MIR-20A, AND MIR-204. RESULTS: IN A HIGH-GLUCOSE CONDITION, ZOLEDRONIC ACID TREATMENT SIGNIFICANTLY LOWERED MALAT1 (P < 0.0001) AND MIR-20A (P < 0.0001) EXPRESSION. CONVERSELY, IN A HIGH-GLUCOSE CONDITION, RUNX2, MIR-133, AND MIR-204 EXPRESSIONS WERE FOUND TO BE SIGNIFICANTLY INCREASED IN THE ZOLEDRONIC ACID TREATMENT GROUP AS COMPARED TO NO TREATMENT (ALL P < 0.0001). CONCLUSIONS: IN CONCLUSION, UNDER A HIGH-GLUCOSE ENVIRONMENT, ZOLEDRONIC ACID CAN MODULATE THE EXPRESSION OF THE RUNX2 TRANSCRIPTION FACTOR THROUGH EPIGENETIC REGULATION.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
2 1144  33 CONCOMITANT HETEROCHROMATINISATION AND DOWN-REGULATION OF GENE EXPRESSION UNVEILS EPIGENETIC SILENCING OF RELB IN AN AGGRESSIVE SUBSET OF CHRONIC LYMPHOCYTIC LEUKEMIA IN MALES. BACKGROUND: THE SENSITIVITY OF CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) CELLS TO CURRENT TREATMENTS, BOTH IN VITRO AND IN VIVO, RELIES ON THEIR ABILITY TO ACTIVATE APOPTOTIC DEATH. CLL CELLS RESISTANT TO DNA DAMAGE-INDUCED APOPTOSIS DISPLAY DEREGULATION OF A SPECIFIC SET OF GENES. METHODS: MICROARRAY HYBRIDIZATION (HUMAN GENECHIP, AFFYMETRIX), IMMUNOFLUORESCENT IN SITU LABELING COUPLED WITH VIDEO-MICROSCOPY RECORDING/ANALYSES, CHROMATIN-IMMUNOPRECIPITATION (CHIP), POLYMERASE CHAIN REACTIONS (PCR), REAL-TIME QUANTITATIVE PCR (RT-QPCR) AND BISULFITE GENOME SEQUENCING WERE THE MAIN METHODS APPLIED. STATISTICAL ANALYSES WERE PERFORMED BY APPLYING GCRMA AND SAM ANALYSIS (MICROARRAY DATA) AND STUDENT'S T-TEST OR MANN & WHITNEY'S U-TEST. RESULTS: HEREIN WE SHOW THAT, REMARKABLY, IN A RESISTANT MALE CLL CELLS THE VAST MAJORITY OF GENES WERE DOWN-REGULATED COMPARED WITH SENSITIVE CELLS, WHEREAS THIS WAS NOT THE CASE IN CELLS DERIVED FROM FEMALES. THIS GENE DOWN-REGULATION WAS FOUND TO BE ASSOCIATED WITH AN OVERALL GAIN OF HETEROCHROMATIN AS EVIDENCED BY IMMUNOFLUORESCENT LABELING OF HETEROCHROMATIN PROTEIN 1ALPHA (HP-1), TRIMETHYLATED HISTONE 3 LYSINE 9 (3METH3K9), AND 5-METHYLCYTIDINE (5METC). NOTABLY, 17 GENES WERE FOUND TO BE COMMONLY DEREGULATED IN RESISTANT MALE AND FEMALE CELL SAMPLES. AMONG THESE, RELB WAS IDENTIFIED AS A DISCRIMINATORY CANDIDATE GENE REPRESSED IN THE MALE AND UPREGULATED IN THE FEMALE RESISTANT CELLS. CONCLUSION: THE MOLECULAR DEFECTS IN THE SILENCING OF RELB INVOLVE AN INCREASE IN H3K9- BUT NOT CPG-ISLAND METHYLATION IN THE PROMOTER REGIONS. INCREASE IN ACETYL-H3 IN RESISTANT FEMALE BUT NOT MALE CLL SAMPLES AS WELL AS A DECREASE OF TOTAL CELLULAR LEVEL OF RELB AFTER AN INHIBITION OF HISTONE DEACETYLASE (HDAC) BY TRICHOSTATIN A (TSA), FURTHER EMPHASIZE THE ROLE OF EPIGENETIC MODIFICATIONS WHICH COULD DISCRIMINATE TWO CLL SUBSETS. TOGETHER, THESE RESULTS HIGHLIGHTED THE EPIGENETIC RELB SILENCING AS A NEW MARKER OF THE PROGRESSIVE DISEASE IN MALES.	2010	
                                                                                 
3 3473  27 IDENTIFICATION OF A NOVEL, METHYLATION-DEPENDENT, RUNX2 REGULATORY REGION ASSOCIATED WITH OSTEOARTHRITIS RISK. OSTEOARTHRITIS (OA) IS A COMMON, MULTIFACTORIAL AND POLYGENIC SKELETAL DISEASE THAT, IN ITS SEVEREST FORM, REQUIRES JOINT REPLACEMENT SURGERY TO RESTORE MOBILITY AND TO RELIEVE CHRONIC PAIN. USING TISSUES FROM THE ARTICULATING JOINTS OF 260 PATIENTS WITH OA AND A RANGE OF IN VITRO EXPERIMENTS, INCLUDING CRISPR-CAS9, WE HAVE CHARACTERIZED AN INTERGENIC REGULATORY ELEMENT. HERE, GENOTYPE AT AN OA RISK LOCUS CORRELATES WITH DIFFERENTIAL DNA METHYLATION, WITH ALTERED GENE EXPRESSION OF BOTH A TRANSCRIPTIONAL REGULATOR (RUNX2), AND A CHROMATIN REMODELLING PROTEIN (SUPT3H). RUNX2 IS A STRONG CANDIDATE FOR OA SUSCEPTIBILITY, WITH ITS ENCODED PROTEIN BEING ESSENTIAL FOR SKELETOGENESIS AND HEALTHY JOINT FUNCTION. THE OA RISK LOCUS INCLUDES SINGLE NUCLEOTIDE POLYMORPHISMS (SNPS) LOCATED WITHIN AND FLANKING THE DIFFERENTIALLY METHYLATED REGION (DMR). THE OA ASSOCIATION SNP, RS10948172, DEMONSTRATES PARTICULARLY STRONG CORRELATION WITH METHYLATION, AND TWO INTERGENIC SNPS FALLING WITHIN THE DMR (RS62435998 AND RS62435999) DEMONSTRATE GENETIC AND EPIGENETIC EFFECTS ON THE REGULATORY ACTIVITY OF THIS REGION. WE THEREFORE POSIT THAT THE OA SIGNAL MEDIATES ITS EFFECT BY MODULATING THE METHYLATION OF THE REGULATORY ELEMENT, WHICH THEN IMPACTS ON GENE EXPRESSION, WITH RUNX2 BEING THE PRINCIPAL TARGET. OUR STUDY HIGHLIGHTS THE INTERPLAY BETWEEN DNA METHYLATION, OA GENETIC RISK AND THE DOWNSTREAM REGULATION OF GENES CRITICAL TO NORMAL JOINT FUNCTION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
4 5273  39 PROMOTER METHYLATION AND BDNF AND DAT1 GENE EXPRESSION PROFILES IN PATIENTS WITH DRUG ADDICTION. BACKGROUND: DRUG ADDICTION IS A BRAIN DISORDER THAT HAS NEGATIVE CONSEQUENCES FOR INDIVIDUALS AND SOCIETY. ADDICTIONS ARE CHRONIC RELAPSING DISEASES OF THE BRAIN THAT ARE CAUSED BY DIRECT DRUG-INDUCED EFFECTS AND PERSEVERING NEUROADAPTATIONS AT THE EPIGENETIC, NEUROPEPTIDE AND NEUROTRANSMITTER LEVELS. BECAUSE THE DOPAMINERGIC SYSTEM HAS A SIGNIFICANT ROLE IN DRUG ABUSE, THE PURPOSE OF THIS STUDY WAS TO ANALYZE THE METHYLATION AND EXPRESSION PROFILE OF BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) AND DOPAMINE TRANSPORTER (DAT1) GENES IN INDIVIDUALS WITH DRUG ADDICTION. MATERIALS AND METHODS: BDNF AND DAT1 PROMOTER METHYLATION WERE INVESTIGATED WITH A METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN BLOOD SAMPLES FROM 75 INDIVIDUALS WITH DRUG ADDICTION AND 65 HEALTHY CONTROLS. THE EXPRESSION LEVELS OF BDNF AND DAT1 WERE ASSESSED IN 12 MRNA SAMPLES FROM THE BLOOD OF PATIENTS AND COMPARED TO THE SAMPLES OF HEALTHY CONTROLS (N = 12) WITH REAL-TIME QUANTITATIVE REVERSE TRANSCRIPTION PCR. RESULTS: NO SIGNIFICANT DIFFERENCES WERE FOUND IN THE METHYLATION OF BDNF AND DAT1 BETWEEN PATIENTS AND CONTROLS, BUT THE RELATIVE LEVELS OF EXPRESSION OF BDNF AND DAT1 MRNA DIFFERED SIGNIFICANTLY IN THE PATIENTS COMPARED TO CONTROLS (P < 0.0001). CONCLUSION: THESE RESULTS SHOWED THAT THE METHYLATION STATUS OF THE BDNF AND DAT1 GENES HAD NO SIGNIFICANT FUNCTION IN THE PROCESSES OF DRUG ADDICTION.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
5 1297  37 DECREASED MRNA EXPRESSION LEVELS OF DNA METHYLTRANSFERASES TYPE 1 AND 3A IN SYSTEMIC LUPUS ERYTHEMATOSUS. OBJECTIVES: SYSTEMIC LUPUS ERYTHEMATOSUS (SLE) IS A CHRONIC RELAPSING AUTOIMMUNE DISEASE CHARACTERIZED BY THE PRESENCE OF AUTOANTIBODIES DIRECTED AGAINST NUCLEAR ANTIGENS AND BY CHRONIC INFLAMMATION. ALTHOUGH THE ETIOLOGY OF SLE REMAINS UNCLEAR, THE INFLUENCE OF ENVIRONMENT FACTORS, WHICH IS LARGELY REFLECTED BY THE EPIGENETIC MECHANISMS, WITH DNA METHYLATION CHANGES IN PARTICULAR, IS GENERALLY CONSIDERED AS MAIN PLAYERS IN THE PATHOGENESIS OF SLE. WE STUDIED DNA METHYLTRANSFERASES' (DNMTS) TYPE 1, 3A AND 3B TRANSCRIPT LEVELS IN PERIPHERAL BLOOD MONONUCLEAR CELLS FROM PATIENTS DIAGNOSED WITH SYSTEMIC LUPUS ERYTHEMATOSUS AND FROM THE HEALTHY CONTROL SUBJECTS. FURTHERMORE, THE ASSOCIATION OF DNMT1, DNMT3A, AND DNMT3B MRNA LEVELS WITH GENDER, AGE, AND MAJOR CLINICAL MANIFESTATIONS WAS ANALYZED. METHODS: PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) WERE ISOLATED FROM 32 SLE PATIENTS AND 40 HEALTHY CONTROLS. REVERSE TRANSCRIPTION AND REAL-TIME QUANTITATIVE POLYMERASE CHAIN REACTION (RT-QPCR) ANALYSES WERE USED TO DETERMINE DNMT1, DNMT3A, AND DNMT3B MRNA EXPRESSION LEVELS. RESULTS: SIGNIFICANTLY LOWER DNMT1 (P = 0.015543) AND DNMT3A (P = 0.003652) TRANSCRIPT LEVELS IN SLE PATIENTS WERE OBSERVED COMPARED WITH HEALTHY CONTROLS. NEVERTHELESS, THE DNMT3B MRNA EXPRESSION LEVELS WERE MARKEDLY LOWER COMPARED WITH DNMT1 AND DNMT3A, BOTH IN PBMCS FROM AFFECTED PATIENTS AND THOSE FROM CONTROL SUBJECTS. FURTHERMORE, THE DNMT1 TRANSCRIPT LEVELS WERE POSITIVELY CORRELATED WITH SLE DISEASE ACTIVITY INDEX (SLEDAI) (R (S) = 0.4087, P = 0.020224), WHILE THE DNMT3A TRANSCRIPT LEVELS WERE NEGATIVELY CORRELATED WITH PATIENTS AGE (R (S) = -0.3765, P = 0.03369). CONCLUSIONS: OUR ANALYSES CONFIRMED THE IMPORTANCE OF EPIGENETIC ALTERATIONS IN SLE ETIOLOGY. MOREOVER, OUR RESULTS SUGGEST THAT THE PRESENCE OF SOME CLINICAL MANIFESTATIONS, SUCH AS PHOTOTOSENSITIVITY AND ARTHRITIS, MIGHT BE ASSOCIATED WITH THE DYSREGULATION OF DNA METHYLTRANSFERASES' MRNA EXPRESSION LEVELS.	2017	
                                                                                                                                                                           
6 3468  33 HYPOXIA-INDUCED DNA HYPERMETHYLATION IN HUMAN PULMONARY FIBROBLASTS IS ASSOCIATED WITH THY-1 PROMOTER METHYLATION AND THE DEVELOPMENT OF A PRO-FIBROTIC PHENOTYPE. BACKGROUND: PULMONARY FIBROSIS IS A DEBILITATING AND LETHAL DISEASE WITH NO EFFECTIVE TREATMENT OPTIONS. UNDERSTANDING THE PATHOLOGICAL PROCESSES AT PLAY WILL DIRECT THE APPLICATION OF NOVEL THERAPEUTIC AVENUES. HYPOXIA HAS BEEN IMPLICATED IN THE PATHOGENESIS OF PULMONARY FIBROSIS YET THE PRECISE MECHANISM BY WHICH IT CONTRIBUTES TO DISEASE PROGRESSION REMAINS TO BE FULLY ELUCIDATED. IT HAS BEEN SHOWN THAT CHRONIC HYPOXIA CAN ALTER DNA METHYLATION PATTERNS IN TUMOUR-DERIVED CELL LINES. THIS EPIGENETIC ALTERATION CAN INDUCE CHANGES IN CELLULAR PHENOTYPE WITH PROMOTER METHYLATION BEING ASSOCIATED WITH GENE SILENCING. OF PARTICULAR RELEVANCE TO IDIOPATHIC PULMONARY FIBROSIS (IPF) IS THE OBSERVATION THAT THY-1 PROMOTER METHYLATION IS ASSOCIATED WITH A MYOFIBROBLAST PHENOTYPE WHERE LOSS OF THY-1 OCCURS ALONGSIDE INCREASED ALPHA SMOOTH MUSCLE ACTIN (ALPHA-SMA) EXPRESSION. THE INITIAL AIM OF THIS STUDY WAS TO DETERMINE WHETHER HYPOXIA REGULATES DNA METHYLATION IN NORMAL HUMAN LUNG FIBROBLASTS (CCD19LU). AS IT HAS BEEN REPORTED THAT HYPOXIA SUPPRESSES THY-1 EXPRESSION DURING LUNG DEVELOPMENT WE ALSO STUDIED THE EFFECT OF HYPOXIA ON THY-1 PROMOTER METHYLATION AND GENE EXPRESSION. METHODS: CCD19LU WERE GROWN FOR UP TO 8 DAYS IN HYPOXIA AND ASSESSED FOR GLOBAL CHANGES IN DNA METHYLATION USING FLOW CYTOMETRY. REAL-TIME PCR WAS USED TO QUANTIFY EXPRESSION OF THY-1, ALPHA-SMA, COLLAGEN I AND III. GENOMIC DNA WAS BISULPHITE TREATED AND METHYLATION SPECIFIC PCR (MSPCR) WAS USED TO EXAMINE THE METHYLATION STATUS OF THE THY-1 PROMOTER. RESULTS: SIGNIFICANT GLOBAL HYPERMETHYLATION WAS DETECTED IN HYPOXIC FIBROBLASTS RELATIVE TO NORMOXIC CONTROLS AND WAS ACCOMPANIED BY INCREASED EXPRESSION OF MYOFIBROBLAST MARKERS. THY-1 MRNA EXPRESSION WAS SUPPRESSED IN HYPOXIC CELLS, WHICH WAS RESTORED WITH THE DEMETHYLATING AGENT 5-AZA-2'-DEOXYCYTIDINE. MSPCR REVEALED THAT THY-1 BECAME METHYLATED FOLLOWING FIBROBLAST EXPOSURE TO 1% O2. CONCLUSION: THESE DATA SUGGEST THAT GLOBAL AND GENE-SPECIFIC CHANGES IN DNA METHYLATION MAY PLAY AN IMPORTANT ROLE IN FIBROBLAST FUNCTION IN HYPOXIA.	2012	

7 6794  32 [EFFECT OF BENZO(A)PYRENE ON THE EXPRESSION OF AHR-REGULATED MICRORNA IN FEMALE AND MALE RAT LUNGS]. SMOKING IS THE MAIN RISK FACTOR FOR LUNG CANCER, MAINLY DUE TO PRESENCE OF NITROSAMINES AND POLYCYCLIC AROMATIC HYDROCARBONS, INCLUDING BENZO[A]PYRENE (BP) IN TOBACCO SMOKE COMPOSITION. THE GENOTOXIC EFFECT OF BP IS BASED ON THE HIGH DNA-BINDING ABILITY OF ITS METABOLITES, WHILE THE EPIGENETIC EFFECTS ARE MEDIATED BY A CHANGE IN THE EXPRESSION OF CANCER RELATED GENES OR REGULATORY RNAS. IT HAS BEEN SHOWN THAT WOMEN HAVE A HIGHER RISK TO DEVELOP LUNG CANCER UPON SMOKING RATHER THAN MEN. WE HYPOTHESIZED THAT CROSSTALK BETWEEN SIGNALING PATHWAYS ACTIVATED BY BP AND ESTROGENS COULD UNDERLIE THE SEX-DEPENDENT DIFFERENCES IN MIRNAS EXPRESSION. TO TEST THIS HYPOTHESIS, MALE AND FEMALE RATS WERE SUBJECTED TO SHORT-TERM OR LONG-TERM BP EXPOSURE. USING IN SILICO ANALYSIS, MIRNAS CONTAINING THE ER- AND AHR-BINDING SITES IN THE PROMOTERS OF THE GENES (OR HOST GENES) WERE SELECTED. DURING CHRONIC EXPOSURE OF BP THE EXPRESSION OF MIR-22-3P, -29A-3P, -126A-3P, -193B-5P IN THE LUNGS OF MALE RATS WERE SIGNIFICANTLY INCREASED, WHILE THE LEVEL OF MIRNA-483-3P WERE DECREASED. EXPRESSION OF MIRNA-483-3P WAS UP-REGULATED DURING CHRONIC BP EXPOSURE IN THE LUNGS OF FEMALE RATS AND THE LEVELS OF OTHER STUDIED MIRNAS WERE UNCHANGED. IN TURN, CHANGES IN THE EXPRESSION OF MIRNAS WERE FOLLOWED BY CHANGES IN THE EXPRESSION OF THEIR TARGET GENES, INCLUDING PTEN, EMP2, IGF1, ITGA6, SLC34A2, AND THE OBSERVED CHANGES IN FEMALE AND MALE RAT LUNGS WERE VARIED. THUS, OUR RESULTS SUGGEST THAT SEX-DEPENDENT EPIGENETIC EFFECTS OF BP MAY BE BASED ON DIFFERENT EXPRESSION OF AHR- AND ER- REGULATED MIRNAS.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
8 2024  35 EPIGENETIC CHANGES CAUSED BY DIABETES AND THEIR POTENTIAL ROLE IN THE DEVELOPMENT OF PERIODONTITIS. AIMS/INTRODUCTION: PERIODONTAL DISEASE, A CHRONIC INFLAMMATION INDUCED BY BACTERIA, IS CLOSELY LINKED WITH DIABETES MELLITUS. MANY COMPLICATIONS ASSOCIATED WITH DIABETES ARE RELATED TO EPIGENETIC CHANGES. HOWEVER, THE EXACT EPIGENETIC CHANGES WHEREBY DIABETES AFFECTS PERIODONTAL DISEASE REMAIN LARGELY UNKNOWN. THUS, WE SOUGHT TO INVESTIGATE THE ROLE OF DIABETES-DEPENDENT EPIGENETIC CHANGES OF GINGIVAL TISSUE IN THE SUSCEPTIBILITY TO PERIODONTAL DISEASE. MATERIALS AND METHODS: WE STUDIED THE EFFECT OF STREPTOZOTOCIN-INDUCED DIABETES IN MINIPIGS ON GINGIVAL MORPHOLOGICAL AND EPIGENETIC TISSUE CHANGES. ACCORDINGLY, WE RANDOMLY DIVIDED SIX MINIPIGS INTO TWO GROUPS: STREPTOZOTOCIN-INDUCED DIABETES GROUP, N = 3; AND NON-DIABETES HEALTHY CONTROL GROUP, N = 3. AFTER 85 DAYS, ALL ANIMALS WERE KILLED, AND GINGIVAL TISSUE WAS COLLECTED FOR HISTOLOGY, DEOXYRIBONUCLEIC ACID METHYLATION ANALYSIS AND IMMUNOHISTOCHEMISTRY. RESULTS: A DIABETES MELLITUS MODEL WAS SUCCESSFULLY CREATED, AS EVIDENCED BY SIGNIFICANTLY INCREASED BLOOD GLUCOSE LEVELS, REDUCTION OF PANCREATIC INSULIN-PRODUCING BETA-CELLS AND HISTOPATHOLOGICAL CHANGES IN THE KIDNEYS. THE GINGIVAL TISSUES IN THE DIABETES GROUP PRESENTED ACANTHOSIS OF BOTH GINGIVAL SQUAMOUS EPITHELIUM AND SULCULAR/JUNCTIONAL EPITHELIUM, AND A SIGNIFICANT REDUCTION IN THE NUMBER AND LENGTH OF RETE PEGS. DEOXYRIBONUCLEIC ACID METHYLATION ANALYSIS SHOWED A TOTAL OF 1,163 AFFECTED GENES, OF WHICH 599 AND 564 WERE SIGNIFICANTLY HYPERMETHYLATED AND HYPOMETHYLATED, RESPECTIVELY. IMMUNOHISTOCHEMISTRY STAINING SHOWED THAT THE HYPOMETHYLATED GENES - TUMOR NECROSIS FACTOR-ALPHA AND INTERLEUKIN-6 - WERE POSITIVELY EXPRESSED UNDER THE JUNCTIONAL EPITHELIUM AREA IN THE DIABETES GROUP. CONCLUSIONS: DIABETES MELLITUS INDUCES MORPHOLOGICAL AND EPIGENETIC CHANGES IN PERIODONTAL TISSUE, WHICH MIGHT CONTRIBUTE TO THE INCREASED SUSCEPTIBILITY OF PERIODONTAL DISEASES IN PATIENTS WITH DIABETES.	2021	
                                                                                                                                                                                                                             
9 1504  29 DNA METHYLATION AND HISTONE DEACETYLATION REGULATING INSULIN SENSITIVITY DUE TO CHRONIC COLD EXPOSURE. IN THIS STUDY, WE INVESTIGATED THE CAUSAL RELATIONSHIP BETWEEN CHRONIC COLD EXPOSURE AND INSULIN RESISTANCE AND THE MECHANISMS OF HOW DNA METHYLATION AND HISTONE DEACETYLATION REGULATE COLD-REDUCED INSULIN RESISTANCE. 46 ADULT MALE MICE FROM POSTNATAL DAY 90-180 WERE RANDOMLY ASSIGNED TO CONTROL GROUP AND COLD-EXPOSURE GROUP. MICE IN COLD-EXPOSURE GROUP WERE PLACED AT TEMPERATURE FROM -1 TO 4 DEGREES C FOR 30 DAYS TO MIMIC CHRONIC COLD ENVIRONMENT. THEN, FASTING BLOOD GLUCOSE, BLOOD INSULIN LEVEL AND INSULIN RESISTANCE INDEX WERE MEASURED WITH ENZYMATIC METHODS. IMMUNOFLUORESCENT LABELING WAS CARRIED OUT TO VISUALIZE THE INSULIN RECEPTOR SUBSTRATE 2 (IRS2), OBESE RECEPTOR (OB-R, A LEPTIN RECEPTOR), VOLTAGE-DEPENDENT ANION CHANNEL PROTEIN 1 (VDAC1), CYTOCHROME C (CYTC), 5-METHYLCYTOSINE (5-MC) POSITIVE CELLS IN HIPPOCAMPAL CA1 AREA. FURTHERMORE, THE EXPRESSIONS OF SOME PROTEINS MENTIONED ABOVE WERE DETECTED WITH WESTERN BLOT. THE RESULTS SHOWED: 1 IN CIRCLE CHRONIC COLD EXPOSURE COULD REDUCE THE INSULIN RESISTANCE INDEX (P < 0.01) AND INCREASE THE NUMBER OF IRS2 POSITIVE CELLS AND OB-R POSITIVE CELLS IN HIPPOCAMPUS (P < 0.01). 2 IN CIRCLE THE EXPRESSIONS OF MITOCHONDRIAL ENERGY-RELATIVE PROTEINS, VDAC1 AND CYTC, WERE HIGHER IN COLD-EXPOSURE GROUP THAN IN CONTROL GROUP WITH BOTH IMMUNOHISTOCHEMICAL STAINING AND WESTERN BLOT (P < 0.01). 3 IN CIRCLE CHRONIC COLD EXPOSURE INCREASED DNA METHYLATION AND HISTONE DEACETYLATION IN THE PYRAMIDAL CELLS OF CA1 AREA AND LED TO AN INCREASE IN THE EXPRESSION OF HISTONE DEACETYLASE 1 (HDAC1) AND DNA METHYLATION RELATIVE ENZYMES (P < 0.01). IN CONCLUSION, CHRONIC COLD EXPOSURE CAN IMPROVE INSULIN SENSITIVITY, WITH THE INVOLVEMENT OF DNA METHYLATION, HISTONE DEACETYLATION AND THE REGULATION OF MITOCHONDRIAL ENERGY METABOLISM. THESE EPIGENETIC MODIFICATIONS PROBABLY FORM THE BASIC MECHANISM OF COLD-REDUCED INSULIN RESISTANCE.	2017	
                                                                                                                                                                                                                                                                 
10 5841  27 STRUCTURAL CHROMATIN ALTERATIONS IN PERIPHERAL BLOOD LEUKOCYTES OF ALCOHOL-DEPENDENT INDIVIDUALS DURING DETOXIFICATION THERAPY. BACKGROUND/AIM: THE AIM OF THIS STUDY WAS TO INVESTIGATE THE STATE OF CHROMATIN CONDENSATION IN PERIPHERAL BLOOD LEUKOCYTES OF ALCOHOLICS, DURING THE EARLY DETOXIFICATION PERIOD, IN ORDER TO HIGHLIGHT STRUCTURAL MODIFICATIONS, INDICATING EPIGENETIC MECHANISMS REGULATED BY ALCOHOL. MATERIALS AND METHODS: BLOOD SAMPLES WERE OBTAINED FROM ALCOHOLIC PATIENTS, WHO WERE ADMITTED FOR DETOXIFICATION ON AN INPATIENT BASIS, AND FROM HEALTHY CONTROLS. THE LEVEL OF CONDENSED HETEROCHROMATIN AND DE-CONDENSED EUCHROMATIN WERE DETECTED THROUGH THE RATIO OF LYSINE TO ARGININE RESIDUES, BY THE APPLICATION OF THE AMMONIACAL SILVER REACTION (ASR) STAINING ON LEUKOCYTE PELLETS, AND THROUGH IMMUNOHISTOCHEMICAL LOCALIZATION OF HISTONE H1 ON PERIPHERAL BLOOD SMEARS. RESULTS: LYMPHOCYTES AND NEUTROPHILS WITH RELAXED DE-CONDENSED CHROMATIN WERE FOUND, INDICATING A MORE REACTIVE GENOME IN ALCOHOLICS, EVEN AT THE STAGE OF DETOXIFICATION. CONCLUSION: THE RESULTS UNDERLINE THE IMPORTANCE OF CHROMATIN STRUCTURE OF LEUKOCYTES AS A SENSITIVE, PERIPHERAL, BIOLOGICAL MARKER FOR EPIGENETIC STUDIES IN LIVING CHRONIC ALCOHOLICS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
11  658  24 BLOOD DNA METHYLATION PREDICTS DIABETIC KIDNEY DISEASE PROGRESSION IN HIGH FAT DIET-FED MICE. DIABETIC KIDNEY DISEASE (DKD) PROGRESSES AT DIFFERENT RATES AMONG PATIENTS WITH TYPE 2 DIABETES MELLITUS (T2D). EARLY IDENTIFICATION OF PATIENTS WITH A HIGHER RISK OF DKD PROGRESSION IS ESSENTIAL TO IMPROVE PROGNOSIS. EPIGENETIC MODIFICATIONS, PARTICULARLY DNA METHYLATION, HAVE BEEN INDEPENDENTLY IMPLICATED IN T2D AND CHRONIC KIDNEY DISEASE. THE CURRENT STUDY AIMED TO DETERMINE CHANGES IN BLOOD DNA METHYLATION THAT REFLECTS AND PREDICTS DKD PROGRESSION. C57BL/6 MICE WERE FED A HIGH-FAT DIET (HFD) FROM WEANING AND SUBCLASSIFIED INTO TWO GROUPS, HFD-1 AND HFD-2, ACCORDING TO URINARY KIDNEY INJURY MARKER KIM-1/CREATININE RATIOS (LOW VS. HIGH) AND HISTOLOGICAL ABNORMALITIES (MILD-MODERATE VS. ADVANCED). DNA METHYLATION PROFILES WERE DETERMINED BY REDUCED REPRESENTATIVE BISULFIDE SEQUENCING (RRBS). OUR RESULTS CONFIRMED EARLY AND ESTABLISHED DKD AT WEEK 9 AND WEEK 32, RESPECTIVELY. AT WEEK 32, ADVANCED KIDNEY INJURY WAS ASSOCIATED WITH DYSREGULATION OF METHYLATION AND DEMETHYLATION ENZYMES IN THE KIDNEY. BLOOD RRBS REVEALED 579 AND 203 DIFFERENTIALLY METHYLATED SITES (DMS) BETWEEN HFD-1 AND HFD-2 ANIMALS AT WEEK 32 AND WEEK 9, RESPECTIVELY, AMONG WHICH 11 WERE COMMON. THE DMS IN BLOOD AND KIDNEY AT WEEK 32 WERE BOTH RELATED TO ORGAN DEVELOPMENT, NEUROGENESIS, CELL JUNCTION, AND WNT SIGNALLING, WHILE THE DMS IN BLOOD AT WEEK 9 SUGGESTED A SPECIFIC ENRICHMENT OF KIDNEY DEVELOPMENT PROCESSES. IN CONCLUSION, OUR DATA STRONGLY SUPPORT THE IMPLICATION OF EARLY BLOOD DNA METHYLATION MODIFICATIONS AND DKD PROGRESSION IN T2D THAT COULD BE USED TO IMPROVE THE DISEASE'S PROGNOSTICATION.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
12 1729  30 DYSREGULATION OF MIR-155 EXPRESSION IN PROFESSIONAL MIXED MARTIAL ARTS (MMA) FIGHTERS. PSYCHOLOGICAL AND PHYSICAL STRESS CAN INDUCE DYSREGULATION OF GENE EXPRESSION VIA CHANGES IN DNA METHYLATION AND MICRORNA (MIRNA) EXPRESSION. SUCH EPIGENETIC MODIFICATIONS ARE YET TO BE INVESTIGATED IN PROFESSIONAL MIXED MARTIAL ARTS (MMA) FIGHTERS SUBJECT TO HIGHLY STRESSFUL TRAINING INVOLVING REPETITIVE HEAD IMPACTS. THIS STUDY EXAMINED DIFFERENCES IN DNA METHYLATION AND MIRNA EXPRESSION IN ELITE MMA FIGHTERS COMPARED TO ACTIVE CONTROLS. GLOBAL METHYLATION DIFFERENCES BETWEEN GROUPS WERE ASSESSED VIA A LINE-1 ASSAY. AT THE SAME TIME, PCR ARRAYS WERE USED TO ESTIMATE DIFFERENTIAL EXPRESSION IN SAMPLES OF 21 FIGHTERS AND 15 CONTROLS FOR 192 DIFFERENT MIRNAS ASSOCIATED WITH INFLAMMATORY DISEASES. AN INDEPENDENT-SAMPLES T-TEST FOUND NO SIGNIFICANT DIFFERENCE IN LINE-1 METHYLATION BETWEEN GROUPS. HOWEVER, AN INDEPENDENT-SAMPLES MANN-WHITNEY U TEST REVEALED A SIGNIFICANT UPREGULATION IN THE EXPRESSION OF MIR-155 IN MMA FIGHTER PLASMA. SINCE MIR-155 HAS BEEN RECOGNIZED AS AN IMPORTANT REGULATOR OF NEUROINFLAMMATION, THIS DYSREGULATION SUGGESTS A POSSIBLE EPIGENETIC MECHANISM RESPONSIBLE FOR CHRONIC INFLAMMATION ASSOCIATED WITH PROFESSIONAL-LEVEL MMA TRAINING. CONSISTENT WITH OTHER PUBLISHED WORKS, THIS STUDY HIGHLIGHTS THE POTENTIAL OF MIR-155 NOT ONLY AS A BIOMARKER FOR MONITORING LONG-TERM HEALTH RISKS LINKED TO HEAD TRAUMA BUT ALSO AS A TARGET TO REMEDIATE THE IMPACT OF CHRONIC NEUROINFLAMMATION.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
13 3866  34 JHDM1D AND HDAC1-3 MRNA EXPRESSION LEVELS IN PERIPHERAL BLOOD MONONUCLEAR CELLS OF PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS. BACKGROUND: SYSTEMIC LUPUS ERYTHEMATOSUS (SLE) IS A CHRONIC RELAPSING AUTOIMMUNE DISEASE CHARACTERIZED BY PRODUCTION OF AUTOANTIBODIES AGAINST A SERIES OF NUCLEAR ANTIGENS AND BY CHRONIC INFLAMMATION. THE ETIOLOGY OF SLE IS THE RESULT OF INTERACTIONS BETWEEN GENETIC, EPIGENETIC, HORMONAL, AND ENVIRONMENTAL FACTORS. CHANGES IN HISTONE ACETYLATION AND METHYLATION CONTRIBUTE TO STRUCTURAL CHROMATIN MODIFICATIONS. OBJECTIVE: WE STUDIED THE HISTONE DEMETHYLASE JHDM1D AND HISTONE DEACETYLASES HDAC1, HDAC2, AND HDAC3 TRANSCRIPT LEVELS IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) FROM PATIENTS DIAGNOSED WITH SYSTEMIC LUPUS ERYTHEMATOSUS (SLE). FURTHERMORE, THE ASSOCIATION OF JHDM1D, HDAC1, HDAC2, AND HDAC3 TRANSCRIPT LEVELS WITH GENDER, AGE, AND MAJOR CLINICAL MANIFESTATIONS WERE ANALYZED. MATERIALS AND METHODS: REAL-TIME QUANTITATIVE POLYMERASE CHAIN REACTION (RQ-PCR) ANALYSIS WAS USED TO DETERMINE JHDM1D, HDAC1, HDAC2, AND HDAC3 MRNA EXPRESSION LEVELS IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) FROM 30 PATIENTS WITH SLE AND 36 HEALTHY CONTROLS. RESULTS: SIGNIFICANTLY LOWER HDAC2 TRANSCRIPT LEVELS (P = 0.006785) AND SIGNIFICANTLY HIGHER JHDM1D (P = 0.0000002) AND HDAC1 (P = 0.010581) TRANSCRIPT LEVELS IN SLE PATIENTS WERE OBSERVED COMPARED WITH HEALTHY CONTROLS. HIGHER JHDM1D MRNA EXPRESSION WAS DETECTED IN ACTIVE SLE PATIENTS WHEN COMPARED WITH INACTIVE PATIENTS (P = 0.005). FURTHERMORE, THE JHDM1D TRANSCRIPT LEVELS WERE POSITIVELY CORRELATED WITH DISEASE ACTIVITY (R(S) = 0.368, P = 0.045), WHILE HDAC2 MRNA EXPRESSION WAS POSITIVELY CORRELATED WITH DISEASE DURATION (R(S) = 0.502, P = 0.0047). CONCLUSION: OUR ANALYSES CONFIRMED THE IMPORTANCE OF EPIGENETIC ALTERATIONS (HISTONE DEMETHYLATION AND ACETYLATION) IN SLE ETIOLOGY. MOREOVER, OUR RESULTS SUGGEST THAT THE PRESENCE OF SOME CLINICAL MANIFESTATIONS, LIKE HEMATOLOGICAL DISEASE AND ANTI-RO ANTIBODY, MIGHT BE ASSOCIATED WITH THE DYSREGULATION OF HISTONE DEMETHYLASE AND DEACETYLASES MRNA EXPRESSION LEVELS.	2015	
                                                                                                                                      
14 4359  34 MIR-422A SUPPRESSES SMAD4 PROTEIN EXPRESSION AND PROMOTES RESISTANCE TO MUSCLE LOSS. BACKGROUND: LOSS OF MUSCLE MASS AND STRENGTH ARE IMPORTANT SEQUELAE OF CHRONIC DISEASE, BUT THE RESPONSE OF INDIVIDUALS IS REMARKABLY VARIABLE, SUGGESTING IMPORTANT GENETIC AND EPIGENETIC MODULATORS OF MUSCLE HOMEOSTASIS. SUCH FACTORS ARE LIKELY TO MODIFY THE ACTIVITY OF PATHWAYS THAT REGULATE WASTING, BUT TO DATE, FEW SUCH FACTORS HAVE BEEN IDENTIFIED. METHODS: THE EFFECT OF MIR-422A ON SMAD4 EXPRESSION AND TRANSFORMING GROWTH FACTOR (TGF)-BETA SIGNALLING WERE DETERMINED BY WESTERN BLOTTING AND LUCIFERASE ASSAY. MIRNA EXPRESSION WAS DETERMINED BY QPCR IN PLASMA AND MUSCLE BIOPSY SAMPLES FROM A CROSS-SECTIONAL STUDY OF PATIENTS WITH CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) AND A LONGITUDINAL STUDY OF PATIENTS UNDERGOING AORTIC SURGERY, WHO WERE SUBSEQUENTLY ADMITTED TO THE INTENSIVE CARE UNIT (ICU). RESULTS: MIR-422A WAS IDENTIFIED, BY A SCREEN, AS A MICRORNA THAT WAS PRESENT IN THE PLASMA OF PATIENTS WITH COPD AND NEGATIVELY ASSOCIATED WITH MUSCLE STRENGTH AS WELL AS BEING READILY DETECTABLE IN THE MUSCLE OF PATIENTS. IN VITRO, MIR-422A SUPPRESSED SMAD4 EXPRESSION AND INHIBITED TGF-BETA AND BONE MORPHOGENETIC PROTEIN-DEPENDENT LUCIFERASE ACTIVITY IN MUSCLE CELLS. IN MALE PATIENTS WITH COPD AND THOSE UNDERGOING AORTIC SURGERY AND ON THE ICU, A MODEL OF ICU-ASSOCIATED MUSCLE WEAKNESS, QUADRICEPS EXPRESSION OF MIR-422A WAS POSITIVELY ASSOCIATED WITH MUSCLE STRENGTH (MAXIMAL VOLUNTARY CONTRACTION R = 0.59, P < 0.001 AND R = 0.51, P = 0.004, FOR COPD AND AORTIC SURGERY, RESPECTIVELY). FURTHERMORE, PRE-SURGERY LEVELS OF MIR-422A WERE INVERSELY ASSOCIATED WITH THE AMOUNT OF MUSCLE THAT WOULD BE LOST IN THE FIRST POST-OPERATIVE WEEK (R = -0.57, P < 0.001). CONCLUSIONS: THESE DATA SUGGEST THAT DIFFERENCES IN MIR-422A EXPRESSION CONTRIBUTE TO THE SUSCEPTIBILITY TO MUSCLE WASTING ASSOCIATED WITH CHRONIC AND ACUTE DISEASE AND THAT AT LEAST PART OF THIS ACTIVITY MAY BE MEDIATED BY REDUCED TGF-BETA SIGNALLING IN SKELETAL MUSCLE.	2018	
                                                                                                                                                                                                                 
15 2395  31 EPIGENETIC REPROGRAMMING IN MIST1(-/-) MICE PREDICTS THE MOLECULAR RESPONSE TO CERULEIN-INDUCED PANCREATITIS. GENE EXPRESSION IS AFFECTED BY MODIFICATIONS TO HISTONE CORE PROTEINS WITHIN CHROMATIN. CHANGES IN THESE MODIFICATIONS, OR EPIGENETIC REPROGRAMMING, CAN DICTATE CELL FATE AND PROMOTE SUSCEPTIBILITY TO DISEASE. THE GOAL OF THIS STUDY WAS TO DETERMINE THE EXTENT OF EPIGENETIC REPROGRAMMING IN RESPONSE TO CHRONIC STRESS THAT OCCURS FOLLOWING ABLATION OF MIST1 (MIST1(-/-) ), WHICH IS REPRESSED IN PANCREATIC DISEASE. CHROMATIN IMMUNOPRECIPITATION FOR TRIMETHYLATION OF LYSINE RESIDUE 4 ON HISTONE 3 (H3K4ME3) IN PURIFIED ACINAR CELLS FROM WILD TYPE AND MIST1(-/-) MICE WAS FOLLOWED BY NEXT GENERATION SEQUENCING (CHIP-SEQ) OR CHIP-QPCR. H3K4ME3-ENRICHED GENES WERE ASSESSED FOR EXPRESSION BY QRT-PCR IN PANCREATIC TISSUE BEFORE AND AFTER INDUCTION OF CERULEIN-INDUCED PANCREATITIS. WHILE MOST OF H3K4ME3-ENRICHMENT IS RESTRICTED TO TRANSCRIPTIONAL START SITES, >25% OF ENRICHMENT SITES ARE FOUND WITHIN, DOWNSTREAM OR BETWEEN ANNOTATED GENES. LESS THAN 10% OF THESE SITES WERE ALTERED IN MIST1(-/-) ACINI, WITH MOST CHANGES IN H3K4ME3 ENRICHMENT NOT REFLECTING ALTERED GENE EXPRESSION. INGENUITY PATHWAY ANALYSIS OF GENES DIFFERENTIALLY-ENRICHED FOR H3K4ME3 REVEALED AN ASSOCIATION WITH PANCREATITIS AND PANCREATIC DUCTAL ADENOCARCINOMA IN MIST1(-/-) TISSUE. MOST OF THESE GENES WERE NOT DIFFERENTIALLY EXPRESSED BUT SEVERAL WERE READILY INDUCED BY ACUTE EXPERIMENTAL PANCREATITIS, WITH SIGNIFICANTLY INCREASED EXPRESSION IN MIST1(-/-) TISSUE RELATIVE TO WILD TYPE MICE. WE SUGGEST THAT THE CHRONIC CELL STRESS OBSERVED IN THE ABSENCE OF MIST1 RESULTS IN EPIGENETIC REPROGRAMMING OF GENES INVOLVED IN PROMOTING PANCREATITIS TO A POISED STATE, THEREBY INCREASING THE SENSITIVITY TO EVENTS THAT PROMOTE DISEASE.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                             
16  782  34 CELL-FREE MICRORNA-148A IS ASSOCIATED WITH RENAL ALLOGRAFT DYSFUNCTION: IMPLICATION FOR BIOMARKER DISCOVERY. BACKGROUND: CHRONIC ALLOGRAFT DYSFUNCTION (CAD), THE FOREMOST CAUSE OF RENAL GRAFT LOSS WORLDWIDE, IS A SERIOUS CHALLENGE FOR MOST OF THE RECIPIENTS. AS THE EPIGENETIC ERA IS EMERGING, EPIGENETIC BIOMARKERS ESPECIALLY MICRORNAS (MIRNAS) MAY REFLECT THE CURRENT STAGE OF THE DISEASE AND PATIENT'S THERAPY RESPONSE. THE CURRENT STUDY INVESTIGATED THE POTENTIAL SIGNIFICANCE OF CIRCULATING MIRNA-148A IN PREDICTING THE RENAL GRAFT FUNCTION. DESIGN AND METHODS: CIRCULATING MIRNAS WERE ISOLATED FROM 53 PLASMA SAMPLES OF RECIPIENTS WITH HISTOLOGICALLY VALIDATED INTERSTITIAL FIBROSIS AND TUBULAR ATROPHY (IFTA, N = 26), AND RECIPIENTS WITH STABLE GRAFT FUNCTION (SGF, N = 27), AND ALSO HEALTHY INDIVIDUALS ( N = 15). THE LEVEL OF MIRNA-148A WAS EVALUATED BY THE QUANTITATIVE POLYMERASE CHAIN REACTION (QPCR) AND CORRELATED WITH CLINICAL AND HISTOLOGICAL PARAMETERS. RESULTS: SIGNIFICANTLY, MIRNA-148A DECREASED IN IFTA COMPARED WITH SGF SUBJECTS (P < 0.001). MIRNA-148A LEVELS INDICATED A SIGNIFICANT ASSOCIATION WITH SERUM CREATININE LEVELS ( R = 0.451, P = 0.021) AND GLOMERULAR FILTRATION RATE ( R = -0.520, P = 0.006). MIRNA-148A EXPRESSION LEVELS COULD DISCRIMINATE IFTA CASES FROM SGF INDIVIDUALS WITH AN AREA UNDER THE CURVE OF 0.89 ( P < 0.001), 97% SENSITIVITY, AND 72% SPECIFICITY. A NUMBER OF PREDICTED TARGETS THAT MIGHT BE INVOLVED IN CAD BY MIRNA-148A WERE PREDICTED. CONCLUSION: PLASMA CELL-FREE MIRNA-148A CORRELATED WITH RENAL FUNCTION AND HISTOLOGICAL GRADES; THEREFORE, IT MAY BE FURTHER INVESTIGATED AS A NOVEL NONINVASIVE MOLECULAR MARKER OF THE PROGRESSION TO IFTA IN RENAL TRANSPLANT RECIPIENTS; MOREOVER, THE EMERGING BIOMARKER MAY BECOME A THERAPEUTIC TARGET IN THE FUTURE CLINIC.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                     
17 1011  27 CIGARETTE SMOKE CONDENSATE INDUCES DIFFERENTIAL EXPRESSION AND PROMOTER METHYLATION PROFILES OF CRITICAL GENES INVOLVED IN LUNG CANCER IN NL-20 LUNG CELLS IN VITRO: SHORT-TERM AND CHRONIC EXPOSURE. ESTABLISHING EARLY DIAGNOSTIC MARKERS OF HARM IS CRITICAL FOR EFFECTIVE PREVENTION PROGRAMS AND REGULATION OF TOBACCO PRODUCTS. THIS STUDY EXAMINED EFFECTS OF CIGARETTE SMOKE CONDENSATE (CSC) ON EXPRESSION AND PROMOTER METHYLATION PROFILE OF CRITICAL GENES (DAPK, ECAD, MGMT, AND RASSF1A) INVOLVED IN LUNG CANCER DEVELOPMENT IN DIFFERENT HUMAN LUNG CELL LINES. NL-20 CELLS WERE TREATED WITH 0.1-100 MUG/ML OF CSC FOR 24 TO 72 HRS FOR SHORT-TERM EXPOSURES. DAPK EXPRESSION OR METHYLATION STATUS WAS NOT SIGNIFICANTLY AFFECTED. HOWEVER, CSC TREATMENT RESULTED IN CHANGES IN EXPRESSION AND PROMOTER METHYLATION PROFILE OF ECAD, MGMT, AND RASSF1A. FOR CHRONIC STUDIES, CELLS WERE EXPOSED TO 1 OR 10 MUG/ML CSC UP TO 28 DAYS. CELLS SHOWED MORPHOLOGICAL CHANGES ASSOCIATED WITH TRANSFORMATION AND CHANGES IN INVASION CAPACITIES AND GLOBAL METHYLATION STATUS. THIS STUDY PROVIDES CRITICAL DATA SUGGESTING THAT EPIGENETIC CHANGES COULD SERVE AS AN EARLY BIOMARKER OF HARM DUE TO EXPOSURE TO CIGARETTE SMOKE.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
18 1272  41 CYTOTOXIC ACTIVITY OF VALPROIC ACID ON PRIMARY CHRONIC LYMPHOCYTIC LEUKEMIA CELLS. BACKGROUND: CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS THE MOST COMMON ADULT LEUKEMIA IN WESTERN CIVILIZATION. THE ACCUMULATION OF CD5+CD19+ B LYMPHOCYTES IN PERIPHERAL BLOOD IS DUE TO A DEFECT IN THE APOPTOTIC PATHWAY RATHER THAN EXCESSIVE PROLIFERATION IN THE BONE MARROW AND LYMPH NODES. DESPITE A NUMBER OF TREATMENTS, CLL REMAINS AN INCURABLE DISEASE. VALPROIC ACID (VPA) ACTIVITY, AS A HISTONE DEACETYLASE INHIBITOR, COULD RESTORE THE EPIGENETIC CHANGES UNDERLYING THE PATHOGENESIS OF CLL AND THUS INDUCE CELL DEATH. OBJECTIVES: IN THE PRESENT STUDY WE HYPOTHESIZED THAT VPA COULD INDUCE CLL PRIMARY CELLS DEATH THROUGH ACTIVATION OF APOPTOSIS. MATERIAL AND METHODS: PERIPHERAL BLOOD SAMPLES WERE OBTAINED FROM 53 CLL PATIENTS. PERIPHERAL BLOOD MONONUCLEAR CELLS WERE ISOLATED THROUGH DENSITY GRADIENT CENTRIFUGATION AND WERE THE SUBJECT OF A 24-HOUR CELL CULTURE WITH 10 MM OF VPA. THE CYTOTOXIC EFFECT OF VPA WAS EVALUATED WITH AN XTT TEST AND THEREAFTER CONFIRMED USING ANNEXIN V-FITC/PI STAINING AND FLOW CYTOMETRY TECHNIQUES. RESULTS: IN THIS STUDY, A MEDIAN VPA CYTOTOXICITY OF 13.88% WITH A RANGE OF 0-54.65% WAS OBSERVED. ANNEXIN V/PI STAINING CONFIRMED THAT THE DEMONSTRATED CYTOTOXICITY WAS CAUSED BY INCREASED APOPTOSIS IN THE VPA TREATED CELLS AS COMPARED TO CONTROL CELLS. STATISTICAL ANALYSIS SHOWED THAT VPA'S EFFECT ON CLL CELLS DEPENDS ON LACTATE DEHYDROGENASE SERUM LEVELS, BUT IS INDEPENDENT OF ALL OTHER PROGNOSTIC MARKERS. CONCLUSIONS: THE RESULTS OF THE PRESENT EXPERIMENTS FOUND THAT VPA AT A CLINICALLY APPLICABLE CONCENTRATION SIGNIFICANTLY INDUCES APOPTOSIS INDEPENDENTLY OF THE DISEASE STAGE AND MIGHT BE A VALUABLE THERAPEUTIC AGENT FOR ALL CLL PATIENTS.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
19 2714  31 EXERCISE-CONDITIONED PLASMA ATTENUATES NUCLEAR CONCENTRATIONS OF DNA METHYLTRANSFERASE 3B IN HUMAN PERIPHERAL BLOOD MONONUCLEAR CELLS. DNA METHYLATION IS MODIFIABLE BY ACUTE AND CHRONIC EXERCISE. DNA METHYLTRANSFERASES (DNMT) CATALYZE THIS PROCESS; HOWEVER, THERE IS A LACK OF LITERATURE CONCERNING THE SPECIFIC MECHANISMS BY WHICH EXERCISE-INDUCED MODIFICATIONS OCCUR. INTERLEUKIN 6 (IL-6) STIMULATION OF VARIOUS CELL LINES HAS BEEN SHOWN TO AUGMENT DNMT EXPRESSION AND NUCLEAR TRANSLOCATION, WHICH SUGGESTS A POSSIBLE PATHWAY BY WHICH EXERCISE IS ABLE TO ELICIT CHANGES IN EPIGENETIC ENZYMES. THE PRESENT STUDY SOUGHT TO ELUCIDATE THE RESPONSE OF THE DE NOVO METHYLTRANSFERASES DNMT3A AND DNMT3B TO CIRCULATORY FACTORS FOUND IN PLASMA ISOLATED FROM WHOLE BLOOD BEFORE AND AFTER 120-MIN OF TREADMILL RUNNING AT AN INTENSITY OF 60% OF INDIVIDUAL VELOCITY AT V O2MAX (VV O2MAX) INTERSPERSED WITH 30-SEC SPRINTS AT 90% OF VV O2MAX EVERY 10-MIN. PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) ISOLATED FROM A RESTING PARTICIPANT WERE INCUBATED WITH PLASMA ISOLATED FROM EXERCISING PARTICIPANTS (N = 10) OR RECOMBINANT IL-6 (RIL-6), FOLLOWED BY NUCLEAR PROTEIN EXTRACTION AND QUANTIFICATION OF DNMT3A AND DNMT3B CONCENTRATIONS. NUCLEAR CONCENTRATIONS OF DNMT3B SIGNIFICANTLY DECREASED FOLLOWING THE EXPERIMENTAL PROTOCOL (P = 0.03), WITH NO CHANGE OBSERVED IN DNMT3A (P = 0.514).VARIOUS CONCENTRATIONS OF RIL-6 CAUSED AN ELEVATION IN BOTH DNMT3A AND DNMT3B NUCLEAR CONCENTRATION COMPARED WITH THE BLANK CONTROL. THE CONFLICTING RESULTS BETWEEN EXERCISING AND RIL-6 CONDITIONS SUGGESTS THAT IL-6 DOES REGULATE DNMT NUCLEAR TRANSPORT, HOWEVER, OTHER PLASMA MEDIATORS MAY ALSO EXERT SIGNIFICANT INFLUENCE ON THE NUCLEAR CONCENTRATIONS OF THESE ENZYMES.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
20 1965  34 EPIGENETIC ALTERATION OF MITOCHONDRIAL BIOGENESIS REGULATORY GENES IN ARSENIC EXPOSED INDIVIDUALS (WITH AND WITHOUT SKIN LESIONS) AND IN SKIN CANCER TISSUES: A CASE CONTROL STUDY. CHRONIC ARSENIC TOXICITY HAS BECOME A GLOBAL CONCERN DUE TO ITS ADVERSE PATHOPHYSIOLOGICAL OUTCOME AND CARCINOGENIC POTENTIAL. IT IS ALREADY ESTABLISHED THAT ARSENIC INDUCED REACTIVE OXYGEN SPECIES ALTERS MITOCHONDRIAL FUNCTIONALITY. MAJOR REGULATORY GENES FOR MITOCHONDRIAL BIOGENESIS, I.E., PGC1ALPHA, TFAM, NRF1AND NRF2 ARE LOCATED IN THE NUCLEUS. AS A RESULT, MITOCHONDRIA-NUCLEUS CROSSTALK IS CRUCIAL FOR PROPER MITOCHONDRIAL FUNCTION. THIS PREVIOUS HYPOTHESIS LED US TO INVESTIGATEINVOLVEMENT OF EPIGENETIC ALTERATION BEHINDENHANCED MITOCHONDRIAL BIOGENESIS IN CHRONIC ARSENIC EXPOSURE. AN EXTENSIVE CASE-CONTROL STUDY WAS CONDUCTED WITH 390 STUDY PARTICIPANTS (UNEXPOSED, EXPOSED WITHOUT SKIN LESION, EXPOSED WITH SKIN LESION AND EXPOSED SKIN TUMOUR) FROM HIGHLY ARSENIC EXPOSED AREAS OFWEST BENGAL, INDIA. METHYLATION SPECIFIC PCRREVEALED SIGNIFICANT PROMOTER HYPOMETHYLATION OFTWO KEY BIOGENESIS REGULATORY GENES, PGC1ALPHAANDTFAM IN ARSENIC EXPOSED INDIVIDUALS AND ALSO IN SKIN TUMOUR TISSUES. LINEAR REGRESSION ANALYSIS INDICATED SIGNIFICANT NEGATIVE CORRELATION BETWEEN URINARY ARSENIC CONCENTRATION AND PROMOTER METHYLATION STATUS. INCREASED EXPRESSION OF BIOGENESIS REGULATORY GENES WASOBTAINED BY QUANTITATIVE REAL-TIME PCR ANALYSIS. MOREOVER, ALTERED MITOCHONDRIAL FUSION-FISSION REGULATORY GENE EXPRESSION WAS ALSO OBSERVED IN SKIN TUMOUR TISSUES. MIR663, HAVING TUMOUR SUPPRESSOR GENE LIKE FUNCTION WAS KNOWN TO BE EPIGENETICALLY REGULATED THROUGH MITOCHONDRIAL RETROGRADE SIGNAL. PROMOTER HYPERMETHYLATION WITH SIGNIFICANTLY DECREASED EXPRESSION OF MIR663 WAS FOUND IN SKIN CANCER TISSUES COMPARED TO NON-CANCEROUS CONTROL TISSUE. IN CONCLUSION, RESULTS INDICATED CRUCIAL ROLE OF EPIGENETIC ALTERATION IN ARSENIC INDUCED MITOCHONDRIAL BIOGENESIS AND ARSENICAL SKIN CARCINOGENESIS FOR THE FIRST TIME. HOWEVER, FURTHER MECHANISTIC STUDIES ARE NECESSARY FOR DETAILED UNDERSTANDING OF MITOCHONDRIA-NUCLEUS CROSSTALK IN ARSENIC PERTURBATION.	2020