1 5568 158 ROLE OF MARIJUANA COMPONENTS ON THE REGENERATIVE ABILITY OF STEM CELLS. STEM CELL THERAPY PROMOTES TISSUE REGENERATION AND WOUND HEALING. EFFORTS HAVE BEEN MADE TO PRIME STEM CELLS TO ENHANCE THEIR REGENERATIVE ABILITIES. CERTAIN MARIJUANA COMPONENTS, NAMELY THE NON-PSYCHOACTIVE CANNABIDIOL (CBD) AND PSYCHOACTIVE TETRAHYDROCANNABINOL (THC), ARE DEFINED AS IMMUNOMODULATORS.(9) WE TEST WHETHER TWO SOURCES OF STEM CELLS, PRIMED WITH CBD OR THC, WOULD DEMONSTRATE IMPROVED REGENERATIVE ABILITIES. HUMAN ADIPOSE-DERIVED STEM CELLS (ASCS) AND BONE MARROW-DERIVED STEM CELLS (BMDSCS), NOT OBTAINED FROM THE SAME INDIVIDUAL, WERE TREATED WITH LOW (300 NM) OR HIGH (3 MUM) CONCENTRATION CBD. PORCINE ASCS AND BMDSCS WERE ISOLATED FROM A SINGLE PIG, AND TREATED WITH EITHER LOW OR HIGH CONCENTRATIONS OF CBD OR THC. TRANSWELL MIGRATION AND MTT PROLIFERATION ASSAYS WERE PERFORMED ON THE HUMAN ASCS AND BMDSCS. ALSO, TRANSWELL MIGRATION ASSAY WAS PERFORMED ON THE PORCINE ASCS AND BMDSCS. FINALLY, A WOUND HEALING SCRATCH ASSAY IN PORCINE PRIMARY FIBROBLASTS (PFS) WAS PERFORMED, CO-CULTURED WITH THE CANNABINOID-TREATED ASCS. CBD PRIMING AT LOW CONCENTRATION INDUCES MIGRATION BY 180% (P < .01) IN PORCINE ASCS, AND BY ONLY 93% (P < .02) IN PORCINE BMDSCS. IN PORCINE STEM CELLS, THC PRIMING AT LOW CONCENTRATION INDUCES MIGRATION BY 91.6% (P < .01) IN ASCS BUT BY ONLY 44.3% (P < .03) IN BMDSCS. COMPARED TO PFS CO-CULTURED WITH UNTREATED ASCS, PFS CO-CULTURED WITH LOW CBD-PRIMED ASCS HAD 75% FASTER WOUND CLOSURE AT 18 HOURS (P < .01). CBD AND THC PRIMING OF ASCS AND BMDSCS, PARTICULARLY AT LOWER DOSES, ENHANCES A NUMBER OF REGENERATIVE PARAMETERS, SUGGESTING THAT THESE MAJOR MARIJUANA COMPONENTS MAY IMPROVE STEM CELL-BASED THERAPIES. SIGNIFICANCE OF THE STUDY: OUR STUDY DEMONSTRATES THAT CANNABINOIDS CAN ENHANCE THE REGENERATIVE CAPACITY OF TWO MAJOR SOURCES OF STEM CELLS, ADIPOSE- AND BONE MARROW-DERIVED, FROM HUMAN AND PORCINE DONORS. STEM CELL ISOLATION AND EXPANSION IS INVASIVE, COSTLY AND TIME CONSUMING. STEM CELLS WITH IMPROVED REGENERATIVE PROPERTIES MAY BE EFFECTIVE IN THE TREATMENT OF ACUTE OR CHRONIC WOUNDS. THIS IS THE FIRST STUDY TO COMPARE THE PRIMING POTENTIAL OF TWO SOURCES OF STEM CELLS FROM THE SAME ANIMAL, WITH THE SAME GENETIC AND EPIGENETIC PROFILE, AS WELL AS THE FIRST TO PRIME WITH THC.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
2 5743  35 SMOKING SUPPRESSES THE THERAPEUTIC POTENTIAL OF ADIPOSE STEM CELLS IN CROHN'S DISEASE PATIENTS THROUGH EPIGENETIC CHANGES. PATIENTS WITH CROHN'S DISEASE (CD) WHO SMOKE ARE KNOWN TO HAVE A WORSE PROGNOSIS THAN NEVER-SMOKERS AND A HIGHER RISK FOR POST-SURGICAL RECURRENCE, WHEREAS PATIENTS WHO QUIT SMOKING AFTER SURGERY HAVE SIGNIFICANTLY LOWER POST-OPERATIVE RECURRENCE. THE HYPOTHESIS WAS THAT SMOKING INDUCES EPIGENETIC CHANGES THAT IMPAIR THE CAPACITY OF ADIPOSE STEM CELLS (ASCS) TO SUPPRESS THE IMMUNE SYSTEM. IT WAS ALSO QUESTIONED WHETHER THIS IMPAIRMENT REMAINS IN EX-SMOKERS WITH CD. ASCS WERE ISOLATED FROM NON-SMOKERS, SMOKERS AND EX-SMOKERS WITH CD AND THEIR INTERACTIONS WITH IMMUNE CELLS WERE STUDIED. THE ASCS FROM BOTH SMOKERS AND EX-SMOKERS PROMOTED MACROPHAGE POLARIZATION TO AN M1 PRO-INFLAMMATORY PHENOTYPE, WERE NOT ABLE TO INHIBIT T- AND B-CELL PROLIFERATION IN VITRO AND ENHANCED THE GENE AND PROTEIN EXPRESSION OF INFLAMMATORY MARKERS INCLUDING INTERLEUKIN-1B. GENOME-WIDE EPIGENETIC ANALYSIS USING TWO DIFFERENT BIOINFORMATIC APPROACHES REVEALED SIGNIFICANT CHANGES IN THE METHYLATION PATTERNS OF GENES THAT ARE CRITICAL FOR WOUND HEALING, IMMUNE AND METABOLIC RESPONSE AND P53-MEDIATED DNA DAMAGE RESPONSE IN ASCS FROM SMOKERS AND EX-SMOKERS WITH CD. IN CONCLUSION, CIGARETTE SMOKING INDUCES A PRO-INFLAMMATORY EPIGENETIC SIGNATURE IN ASCS THAT LIKELY COMPROMISES THEIR THERAPEUTIC POTENTIAL.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
3 1663  46 DOWNREGULATION OF DNA METHYLTRANSFERASE-3A AMELIORATES THE OSTEOGENIC DIFFERENTIATION ABILITY OF ADIPOSE-DERIVED STEM CELLS IN DIABETIC OSTEOPOROSIS VIA WNT/BETA-CATENIN SIGNALING PATHWAY. BACKGROUND: DIABETES-RELATED OSTEOPOROSIS (DOP) IS A CHRONIC DISEASE CAUSED BY THE HIGH GLUCOSE ENVIRONMENT THAT INDUCES A METABOLIC DISORDER OF OSTEOCYTES AND OSTEOBLAST-ASSOCIATED MESENCHYMAL STEM CELLS. THE PROCESSES OF BONE DEFECT REPAIR AND REGENERATION BECOME EXTREMELY DIFFICULT WITH DOP. ADIPOSE-DERIVED STEM CELLS (ASCS), AS SEED CELLS IN BONE TISSUE ENGINEERING TECHNOLOGY, PROVIDE A PROMISING THERAPEUTIC APPROACH FOR BONE REGENERATION IN DOP PATIENTS. THE OSTEOGENIC ABILITY OF ASCS IS LOWER IN A DOP MODEL THAN THAT OF CONTROL ASCS. DNA METHYLATION, AS A MECHANISM OF EPIGENETIC REGULATION, MAY BE INVOLVED IN DNA METHYLATION OF VARIOUS GENES, THEREBY PARTICIPATING IN BIOLOGICAL BEHAVIORS OF VARIOUS CELLS. EMERGING EVIDENCE SUGGESTS THAT INCREASED DNA METHYLATION LEVELS ARE ASSOCIATED WITH ACTIVATION OF WNT/BETA-CATENIN SIGNALING PATHWAY. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE THE INFLUENCE OF THE DIABETIC ENVIRONMENT ON THE OSTEOGENIC POTENTIAL OF ASCS, TO EXPLORE THE ROLE OF DNA METHYLATION ON OSTEOGENIC DIFFERENTIATION OF DOP-ASCS VIA WNT/BETA-CATENIN SIGNALING PATHWAY, AND TO IMPROVE THE OSTEOGENIC DIFFERENTIATION ABILITY OF ASCS WITH DOP. METHODS: DOP-ASCS AND CONTROL ASCS WERE ISOLATED FROM DOP C57BL/6 AND CONTROL MICE, RESPECTIVELY. THE MULTIPOTENCY OF DOP-ASCS WAS CONFIRMED BY ALIZARIN RED-S, OIL RED-O, AND ALCIAN BLUE STAINING. REAL-TIME POLYMERASE CHAIN REACTION (RT-PCR), IMMUNOFLUORESCENCE, AND WESTERN BLOTTING WERE USED TO ANALYZE CHANGES IN MARKERS OF OSTEOGENIC DIFFERENTIATION, DNA METHYLATION, AND WNT/BETA-CATENIN SIGNALING. ALIZARIN RED-S STAINING WAS ALSO USED TO CONFIRM CHANGES IN THE OSTEOGENIC ABILITY. DNMT SMALL INTERFERING RNA (SIRNA), SHRNA-DNMT3A, AND LVRNA-DNMT3A WERE USED TO ASSESS THE ROLE OF DNMT3A IN OSTEOGENIC DIFFERENTIATION OF CONTROL ASCS AND DOP-ASCS. MICRO-COMPUTED TOMOGRAPHY, HEMATOXYLIN AND EOSIN STAINING, AND MASSON STAINING WERE USED TO ANALYZE CHANGES IN THE OSTEOGENIC CAPABILITY WHILE DOWNREGULATING DNMT3A WITH LENTIVIRUS IN DOP MICE IN VIVO. RESULTS: THE PROLIFERATIVE ABILITY OF DOP-ASCS WAS LOWER THAN THAT OF CONTROL ASCS. DOP-ASCS SHOWED A DECREASE IN OSTEOGENIC DIFFERENTIATION CAPACITY, LOWER WNT/BETA-CATENIN SIGNALING PATHWAY ACTIVITY, AND A HIGHER LEVEL OF DNMT3A THAN CONTROL ASCS. WHEN DNMT3A WAS DOWNREGULATED BY SIRNA AND SHRNA, OSTEOGENIC-RELATED FACTORS RUNT-RELATED TRANSCRIPTION FACTOR 2 AND OSTEOPONTIN, AND ACTIVITY OF WNT/BETA-CATENIN SIGNALING PATHWAY WERE INCREASED, WHICH RESCUED THE POOR OSTEOGENIC POTENTIAL OF DOP-ASCS. WHEN DNMT3A WAS UPREGULATED BY LVRNA-DNMT3A, THE OSTEOGENIC ABILITY WAS INHIBITED. THE SAME RESULTS WERE OBTAINED IN VIVO. CONCLUSIONS: DNMT3A SILENCING RESCUES THE NEGATIVE EFFECTS OF DOP ON ASCS AND PROVIDES A POSSIBLE APPROACH FOR BONE TISSUE REGENERATION IN PATIENTS WITH DIABETIC OSTEOPOROSIS.	2022	

4 1980  31 EPIGENETIC ALTERATIONS IN CYTOCHROME P450 OXIDOREDUCTASE (POR) IN SPERM OF RATS EXPOSED TO TETRAHYDROCANNABINOL (THC). AS MARIJUANA LEGALIZATION IS INCREASING, RESEARCH REGARDING POSSIBLE LONG-TERM RISKS FOR USERS AND THEIR OFFSPRING IS NEEDED. LITTLE DATA EXISTS ON EFFECTS OF PATERNAL TETRAHYDROCANNABINOL (THC) EXPOSURE PRIOR TO REPRODUCTION. THIS STUDY DETERMINED IF CHRONIC THC EXPOSURE ALTERS SPERM DNA METHYLATION (DNAM) AND IF SUCH EFFECTS ARE INTERGENERATIONALLY TRANSMITTED. ADULT MALE RATS UNDERWENT ORAL GAVAGE WITH THC OR VEHICLE CONTROL. DIFFERENTIALLY METHYLATED (DM) LOCI IN MOTILE SPERM WERE IDENTIFIED USING REDUCED REPRESENTATION BISULFITE SEQUENCING (RRBS). ANOTHER COHORT WAS INJECTED WITH VEHICLE OR THC, AND SPERM DNAM WAS ANALYZED. FINALLY, THC-EXPOSED AND CONTROL ADULT MALE RATS WERE MATED WITH THC-NAIVE FEMALES. DNAM LEVELS OF TARGET GENES IN BRAIN TISSUES OF THE OFFSPRING WERE DETERMINED BY PYROSEQUENCING. RRBS IDENTIFIED 2,940 DM CPGS MAPPING TO 627 GENES. SIGNIFICANT HYPERMETHYLATION WAS CONFIRMED (P < 0.05) FOLLOWING ORAL THC ADMINISTRATION FOR CYTOCHROME P450 OXIDOREDUCTASE (POR), INVOLVED IN TOXIN PROCESSING AND DISORDERS OF SEXUAL DEVELOPMENT. POR HYPERMETHYLATION WAS NOT OBSERVED AFTER THC INJECTION OR IN THE SUBSEQUENT GENERATION. THESE RESULTS SUPPORT THAT THC ALTERS DNAM IN SPERM AND THAT ROUTE OF EXPOSURE CAN HAVE DIFFERENTIAL EFFECTS. ALTHOUGH WE DID NOT OBSERVE EVIDENCE OF INTERGENERATIONAL TRANSMISSION OF THE DNAM CHANGE, LARGER STUDIES ARE REQUIRED TO DEFINITIVELY EXCLUDE THIS POSSIBILITY.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
5  387  30 AN INDIVIDUALITY OF RESPONSE TO CANNABINOIDS: CHALLENGES IN SAFETY AND EFFICACY OF CANNABIS PRODUCTS. SINCE LEGALIZATION, CANNABIS/MARIJUANA HAS BEEN GAINING CONSIDERABLE ATTENTION AS A FUNCTIONAL INGREDIENT IN FOOD. ?-9 TETRAHYDROCANNABINOL (THC), CANNABIDIOL (CBD), AND OTHER CANNABINOIDS ARE KEY BIOACTIVE COMPOUNDS WITH HEALTH BENEFITS. THE ORAL CONSUMPTION OF CANNABIS TRANSPORTS MUCH LESS HAZARDOUS CHEMICALS THAN SMOKING. NEVERTHELESS, THE RESPONSE TO CANNABIS IS BIPHASICALLY DOSE-DEPENDENT (HORMESIS; A LOW-DOSE STIMULATION AND A HIGH-DOSE INHIBITION) WITH WIDE INDIVIDUALITY IN RESPONSES. THUS, THE EXACT SAME DOSE AND PREPARATION OF CANNABIS MAY BE BENEFICIAL FOR SOME BUT TOXIC TO OTHERS. THE PURPOSE OF THIS REVIEW IS TO HIGHLIGHT THE CONCEPT OF INDIVIDUAL VARIATIONS IN RESPONSE TO CANNABINOIDS, WHICH LEADS TO THE CHALLENGE OF ESTABLISHING STANDARD SAFE DOSES OF CANNABIS PRODUCTS FOR THE GENERAL POPULATION. THE MECHANISMS OF ACTIONS, ACUTE AND CHRONIC TOXICITIES, AND FACTORS AFFECTING RESPONSES TO CANNABIS PRODUCTS ARE UPDATED. BASED ON THE LITERATURE REVIEW, WE FOUND THAT THE RESPONSE TO CANNABIS PRODUCTS DEPENDS ON EXPOSURE FACTORS (DELIVERY ROUTE, DURATION, FREQUENCY, AND INTERACTIONS WITH FOOD AND DRUGS), INDIVIDUAL FACTORS (AGE, SEX), AND SUSCEPTIBILITY FACTORS (GENETIC POLYMORPHISMS OF CANNABINOID RECEPTOR GENE, N-ACYLETHANOLAMINE-HYDROLYZING ENZYMES, THC-METABOLIZING ENZYMES, AND EPIGENETIC REGULATIONS). OWING TO THE INDIVIDUALITY OF RESPONSES, THE SAFEST WAY TO USE CANNABIS-CONTAINING FOOD PRODUCTS IS TO START LOW, GO SLOW, AND STAY LOW.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
6 1519  31 DNA METHYLATION AT ATP11A CG11702988 IS A BIOMARKER OF LUNG DISEASE SEVERITY IN CYSTIC FIBROSIS: A LONGITUDINAL STUDY. CYSTIC FIBROSIS (CF) IS A CHRONIC GENETIC DISEASE THAT MAINLY AFFECTS THE RESPIRATORY AND GASTROINTESTINAL SYSTEMS. NO CURATIVE TREATMENTS ARE AVAILABLE, BUT THE FOLLOW-UP IN SPECIALIZED CENTERS HAS GREATLY IMPROVED THE PATIENT LIFE EXPECTANCY. ROBUST BIOMARKERS ARE REQUIRED TO MONITOR THE DISEASE, GUIDE TREATMENTS, STRATIFY PATIENTS, AND PROVIDE OUTCOME MEASURES IN CLINICAL TRIALS. IN THE PRESENT STUDY, WE OUTLINE A STRATEGY TO SELECT PUTATIVE DNA METHYLATION BIOMARKERS OF LUNG DISEASE SEVERITY IN CYSTIC FIBROSIS PATIENTS. IN THE DISCOVERY STEP, WE SELECTED SEVEN POTENTIAL BIOMARKERS USING A GENOME-WIDE DNA METHYLATION DATASET THAT WE GENERATED IN NASAL EPITHELIAL SAMPLES FROM THE METHYLCF COHORT. IN THE REPLICATION STEP, WE ASSESSED THE SAME BIOMARKERS USING SPUTUM CELL SAMPLES FROM THE METHYLBIOMARK COHORT. OF INTEREST, DNA METHYLATION AT THE CG11702988 SITE (ATP11A GENE) POSITIVELY CORRELATED WITH LUNG FUNCTION AND BMI, AND NEGATIVELY CORRELATED WITH LUNG DISEASE SEVERITY, P. AERUGINOSA CHRONIC INFECTION, AND THE NUMBER OF EXACERBATIONS. THESE RESULTS WERE REPLICATED IN PROSPECTIVE SPUTUM SAMPLES COLLECTED AT FOUR TIME POINTS WITHIN AN 18-MONTH PERIOD AND LONGITUDINALLY. TO CONCLUDE, (I) WE IDENTIFIED A DNA METHYLATION BIOMARKER THAT CORRELATES WITH CF SEVERITY, (II) WE PROVIDED A METHOD TO EASILY ASSESS THIS BIOMARKER, AND (III) WE CARRIED OUT THE FIRST LONGITUDINAL ANALYSIS OF DNA METHYLATION IN CF PATIENTS. THIS NEW EPIGENETIC BIOMARKER COULD BE USED TO STRATIFY CF PATIENTS IN CLINICAL TRIALS.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
7 5722  39 SISTER CHROMATID EXCHANGE AND PROLIFERATIVE RATE INDEX IN THE LONGITUDINAL RISK ASSESSMENT OF OCCUPATIONAL EXPOSURE TO PESTICIDES. AT PRESENT, THERE ARE MORE THAN 1,000 CHEMICALS CLASSIFIED AS PESTICIDES AND MANY REPORTS HAVE SHOWN THAT SOME OF THEM HAVE GENOTOXIC PROPERTIES. IN THE PRESENT LONGITUDINAL STUDY, POSSIBLE GENETIC DAMAGE ON A POPULATION OF WORKERS OCCUPATIONALLY EXPOSED TO A MIXTURE OF PESTICIDES BY USING SISTER CHROMATID EXCHANGE (SCE) ANALYSIS HAS BEEN EVALUATED. AS AN ADDITIONAL CYTOGENETIC PARAMETER, THE PROPORTION OF LYMPHOCYTES THAT UNDERGO ONE, TWO OR THREE CELL DIVISIONS AS WELL AS PROLIFERATIVE RATE INDEX HAVE BEEN DETERMINED. THIS STUDY WAS PERFORMED ON THE EXPOSED GROUP OF WORKERS EMPLOYED IN PESTICIDE PRODUCTION, SIMULTANEOUSLY EXPOSED TO A COMPLEX MIXTURE OF PESTICIDES (ATRAZINE, ALACHLOR, CYANAZINE, 2,4-DICHLOROPHENOXYACETIC ACID, AND MALATHION). THE BLOOD SAMPLES OF THE EXPOSED SUBJECTS WERE COLLECTED IN THREE DIFFERENT PERIODS: BEFORE THE BEGINNING OF THE NEW PESTICIDE PRODUCTION PERIOD, AFTER 8 MONTHS OF EVERYDAY WORK IN THE PESTICIDE PRODUCTION, AND 8 MONTHS AFTER THE REMOVAL OF SUBJECTS OUT OF THE PRODUCTION. IN ALL THREE SAMPLINGS, THE MEAN VALUE OF SCE AND NUMBER OF CELLS WITH HIGH SISTER CHROMATID EXCHANGE FREQUENCY (HFC) IN THE EXPOSED GROUP WAS SIGNIFICANTLY HIGHER IN THE COMPARISON WITH THE CONTROL GROUP. THERE WERE NO DIFFERENCES IN THE PROLIFERATIVE RATE INDEX (PRI) BETWEEN THE CONTROL AND EXPOSED GROUP, REGARDLESS OF THE SAMPLING PERIOD. IN BOTH GROUPS EXAMINED, THE MAJORITY OF LYMPHOCYTES WERE FOUND IN THE SECOND CELL DIVISION, FOLLOWING CULTIVATION. THESE RESULTS SUGGEST THAT THE INCREASE IN THE NUMBER OF SCE FOUND IN THE EXPOSED SUBJECTS IS NOT THE RESULT OF EITHER CYTOTOXIC OR EPIGENETIC ACTION OF PESTICIDE MIXTURE, BUT CHRONIC OCCUPATIONAL EXPOSURE TO MIXTURE OF PESTICIDES.	2002	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
8 4528  21 MULTIGENERATIONAL EFFECTS OF CADMIUM ON THE LIFESPAN AND FERTILITY OF DROSOPHILA MELANOGASTER. ALTHOUGH THE DAMAGE AND TOLERANCE MECHANISMS OF CD STRESS ARE KNOWN, THE DATA ON GENETIC RISK ARE LIMITED. THE AIM OF THIS STUDY WAS TO ASSESS THE CHRONIC TOXICITY OF CD, GENETIC RESPONSES, AND MULTIGENERATIONAL EFFECTS IN FIVE GENERATIONS OF DROSOPHILA MELANOGASTER. FOR EACH GENERATION, LIFESPAN AND FERTILITY WERE STATISTICALLY ANALYSED AND THE EXPRESSION OF APOPTOSIS- (P53 AND CASPASE-3) AND EPIGENESIS-RELATED (DDNMT2 AND DMBD2/3) GENES WAS EXAMINED. LIFESPAN AND FERTILITY SIGNIFICANTLY DECLINED UNDER CD STRESS AND THESE EFFECTS WERE MAINTAINED FOR TWO GENERATIONS AND ONE GENERATION, RESPECTIVELY, WHEN CD STRESS WAS REMOVED. THE EXPRESSION OF P53 AND CASPASE-3 WAS SIGNIFICANTLY UP-REGULATED AFTER EXPOSURE, SUGGESTING THAT APOPTOSIS CONTRIBUTES TO THE RESISTANCE MECHANISM. THEIR ALTERED EXPRESSION WAS RETAINED FOR TWO GENERATIONS. FURTHERMORE, HIGH EXPRESSION OF DDNMT2 AND DMBD2/3 ACCOMPANIED CD EXPOSURE, WHICH WAS PASSED ON TO THREE GENERATIONS, SUGGESTING THAT GENETIC MODIFICATIONS IN APOPTOSIS-RELATED GENES ARE CARRIED TO THE OFFSPRING THROUGH EPIGENETIC REGULATION.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
9 2908  32 GENE EXPRESSION PROFILE OF NORMAL BREAST TISSUE AND BODY MASS INDEX. BACKGROUND: IN HUMAN BREAST, ADIPOSE TISSUE REPRESENTS UP TO 80% OF THE TOTAL VOLUME AND PLAYS A CRITICAL ROLE IN MAMMARY GLAND REMODELING. GIVEN THE EMERGING ROLE OF OBESITY IN BREAST CANCER GROWTH AND DEVELOPMENT, WE EXPLORED THE RELATIONSHIP BETWEEN BODY MASS INDEX (BMI), AS A PROXY OF WOMAN'S OBESITY STATUS, AND THE EXPRESSION IN NORMAL BREAST TISSUE FROM HEALTHY WOMEN OF A SELECTED PANEL OF GENES, KNOWN TO BE INVOLVED IN MAMMARY GLAND HOMEOSTASIS. METHODS: TWO INDEPENDENT PUBLICLY AVAILABLE DATASETS, COMPOSED OF 180 SPECIMENS OF NORMAL BREAST TISSUE FROM REDUCTION MAMMOPLASTY WERE INTERROGATED. DIFFERENTIAL GENE EXPRESSION AMONG BMI CLASSES WAS EVALUATED BY ANOVA, AND PARTIAL CORRELATION COEFFICIENT WAS USED TO ASSAY THE CORRELATION BETWEEN GENES CONTROLLING FOR BMI. RESULTS: DESPITE THE DIFFERENCES IN MICROARRAY PLATFORMS AND ANALYTICAL PROCEDURES, THE TWO DATASETS SHARED A CORE OF 9 GENES DIFFERENTIALLY EXPRESSED IN BMI CLASSES AND SIGNIFICANTLY CORRELATED WITH BMI. FOUR (44%) OF THESE GENES BELONG TO THE FUNCTIONAL CLASS OF CYTOKINES AND CYTOKINE RECEPTORS (IL1R1, IL2RA, IL12A, AND IL12RB2). THE OTHERS BELONG TO THE FUNCTIONAL CLASS OF THE EPIGENETIC REGULATION (MEDAG AND SETD7), SIGNAL TRANSDUCTION (STAT1), CELL ADHESION (ITGAV), AND ENZYMATIC ACTIVITY (STS). CONCLUSIONS: ALTHOUGH EXPLORATORY, PRESENT FINDINGS ARE IN AGREEMENT WITH THE ROLE OF INFLAMMATION MODULATORS IN THE HOMEOSTASIS OF NORMAL BREAST TISSUE AND THE BELIEVE THAT AN INCREASE IN BODY ADIPOSE TISSUE MAY HAVE A POTENTIALLY DANGEROUS LOCAL EFFECT, THROUGH THE INCREASED EXPRESSION OF INFLAMMATION-RELATED GENES AND THE ESTABLISHMENT OF A LOW-GRADE CHRONIC INFLAMMATION.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
10 1272  40 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
11 5894  36 T CELL EPIGENETIC REMODELING AND ACCELERATED EPIGENETIC AGING ARE LINKED TO LONG-TERM IMMUNE ALTERATIONS IN CHILDHOOD CANCER SURVIVORS. BACKGROUND: CANCER TREATMENTS HAVE SUBSTANTIALLY IMPROVED CHILDHOOD CANCER SURVIVAL BUT ARE ACCOMPANIED BY LONG-TERM COMPLICATIONS, NOTABLY CHRONIC INFLAMMATORY DISEASES. WE HYPOTHESIZE THAT CANCER TREATMENTS COULD LEAD TO LONG-TERM EPIGENETIC CHANGES IN IMMUNE CELLS, RESULTING IN INCREASED PREVALENCE OF INFLAMMATORY DISEASES IN CANCER SURVIVORS. RESULTS: TO TEST THIS HYPOTHESIS, WE ESTABLISHED THE EPIGENETIC AND TRANSCRIPTOMIC PROFILES OF IMMUNE CELLS FROM 44 CHILDHOOD CANCER SURVIVORS (CCS, > 16 YEARS OLD) ON FULL REMISSION (> 5 YEARS) WHO HAD RECEIVED CHEMOTHERAPY ALONE OR IN COMBINATION WITH TOTAL BODY IRRADIATION (TBI) AND HEMATOPOIETIC STEM CELL TRANSPLANT (HSCT). WE FOUND THAT MORE THAN 10 YEARS POST-TREATMENT, CCS TREATED WITH TBI/HSCT SHOWED AN ALTERED DNA METHYLATION SIGNATURE IN T CELL, PARTICULARLY AT GENES CONTROLLING IMMUNE AND INFLAMMATORY PROCESSES AND OXIDATIVE STRESS. DNA METHYLATION REMODELING IN T CELL WAS PARTIALLY ASSOCIATED WITH CHRONIC EXPRESSION CHANGES OF NEARBY GENES, INCREASED FREQUENCY OF TYPE 1 CYTOKINE-PRODUCING T CELL, ELEVATED SYSTEMIC LEVELS OF THESE CYTOKINES, AND OVER-ACTIVATION OF RELATED SIGNALING PATHWAYS. SURVIVORS EXPOSED TO TBI/HSCT WERE FURTHER CHARACTERIZED BY AN EPIGENETIC-AGING-SIGNATURE OF T CELL CONSISTENT WITH ACCELERATED EPIGENETIC AGING. TO INVESTIGATE THE POTENTIAL CONTRIBUTION OF IRRADIATION TO THESE CHANGES, WE ESTABLISHED TWO CELL CULTURE MODELS. WE IDENTIFIED THAT RADIATION PARTIALLY RECAPITULATED THE IMMUNE CHANGES OBSERVED IN SURVIVORS THROUGH A BYSTANDER EFFECT THAT COULD BE MEDIATED BY CIRCULATING FACTORS. CONCLUSION: CANCER TREATMENTS, IN PARTICULAR TBI/HSCT, ARE ASSOCIATED WITH LONG-TERM IMMUNE DISTURBANCES. WE PROPOSE THAT EPIGENETIC REMODELING OF IMMUNE CELLS FOLLOWING CANCER THERAPY AUGMENTS INFLAMMATORY- AND AGE-RELATED DISEASES, INCLUDING METABOLIC COMPLICATIONS, IN CHILDHOOD CANCER SURVIVORS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
12 6564  34 TRANSIENT EXPOSURE TO ELEVATED GLUCOSE LEVELS CAUSES PERSISTENT CHANGES IN DERMAL MICROVASCULAR ENDOTHELIAL CELL RESPONSES TO INJURY. BACKGROUND: THE PURPOSE OF THIS STUDY WAS TO DETERMINE WHETHER ELEVATED GLUCOSE CAN INDUCE A DERMAL MICROVASCULAR ENDOTHELIAL CELL METABOLIC MEMORY, THUS AFFECTING ANGIOGENESIS IN THE REPAIR PROCESS OF MAMMALIAN CUTANEOUS WOUND. WE HYPOTHESIZED THAT TRANSIENT ELEVATED GLUCOSE LEVELS CAUSE SUSTAINED ALTERATION OF ENDOTHELIAL CELL RESPONSES TO INJURY AND PERSISTENT EPIGENETIC CHANGES IN GENE EXPRESSION. METHODS: HUMAN DERMAL MICROVASCULAR ENDOTHELIAL CELLS WERE EXPOSED TO EXPERIMENTAL CONDITIONS WITH OR WITHOUT 30 MM D-GLUCOSE. THE CONTROL GROUP WAS MAINTAINED AT 5 MM D-GLUCOSE; WHILE IN THE TRANSIENT GLUCOSE GROUP, AFTER BEING EXPOSED TO 30 MM D-GLUCOSE FOR TWO DAYS, THEN BEING PUT UNDER THE CONTROL CONDITIONS DURING THE EXPERIMENT. BESIDES, IN THE WHOLE PROCESS OF THE EXPERIMENT, THE CHRONIC GLUCOSE GROUP WAS KEPT IN THE CONDITION WITH 30 MM D-GLUCOSE. PROLIFERATION, MIGRATION, TUBE FORMATION, GENE EXPRESSION AND HISTONE METHYLATION WERE ASSESSED FOR INDIVIDUAL CONDITIONS. RESULTS: TRANSIENT ELEVATED GLUCOSE CAUSED SUSTAINED EFFECTS ON ENDOTHELIAL CELL MIGRATION, TUBE FORMATION AND TIMP3 GENE EXPRESSION. THE EFFECTS ON TIMP3 EXPRESSION WERE ASSOCIATED WITH PERSISTENT CHANGES IN HISTONE MODIFICATION AT THE 5' END OF THE TIMP3 GENE, SUGGESTING AN EPIGENETIC EFFECT. CONCLUSIONS: HYPERGLYCEMIA INDUCED METABOLIC MEMORY COULD PROMOTE THE REGULATION OF TIMP3, AND IT CAN BE USED AS A POSSIBLE INNOVATIVE MOLECULAR TARGET FOR THERAPEUTIC INTERVENTION IN THE TREATMENT OF CHRONIC NON-HEALING DIABETIC WOUNDS.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
13 2653  36 EPIGENOTYPING IN PERIPHERAL BLOOD CELL DNA AND BREAST CANCER RISK: A PROOF OF PRINCIPLE STUDY. BACKGROUND: EPIGENETIC CHANGES ARE EMERGING AS ONE OF THE MOST IMPORTANT EVENTS IN CARCINOGENESIS. TWO ALTERATIONS IN THE PATTERN OF DNA METHYLATION IN BREAST CANCER (BC) HAVE BEEN PREVIOUSLY REPORTED; ACTIVE ESTROGEN RECEPTOR-ALPHA (ER-ALPHA) IS ASSOCIATED WITH DECREASED METHYLATION OF ER-ALPHA TARGET (ERT) GENES, AND POLYCOMB GROUP TARGET (PCGT) GENES ARE MORE LIKELY THAN OTHER GENES TO HAVE PROMOTER DNA HYPERMETHYLATION IN CANCER. HOWEVER, WHETHER DNA METHYLATION IN NORMAL UNRELATED CELLS IS ASSOCIATED WITH BC RISK AND WHETHER THESE IMPRINTS CAN BE RELATED TO FACTORS WHICH CAN BE MODIFIED BY THE ENVIRONMENT, IS UNCLEAR. METHODOLOGY/PRINCIPAL FINDINGS: USING QUANTITATIVE METHYLATION ANALYSIS IN A CASE-CONTROL STUDY (N = 1,083) WE FOUND THAT DNA METHYLATION OF PERIPHERAL BLOOD CELL DNA PROVIDES GOOD PREDICTION OF BC RISK. WE ALSO REPORT THAT INVASIVE DUCTAL AND INVASIVE LOBULAR BC IS CHARACTERIZED BY TWO DIFFERENT SETS OF GENES, THE LATTER PARTICULAR BY GENES INVOLVED IN THE DIFFERENTIATION OF THE MESENCHYME (PITX2, TITF1, GDNF AND MYOD1). FINALLY WE DEMONSTRATE THAT ONLY ERT GENES PREDICT ER POSITIVE BC; LACK OF PERIPHERAL BLOOD CELL DNA METHYLATION OF ZNF217 PREDICTED BC INDEPENDENT OF AGE AND FAMILY HISTORY (ODDS RATIO 1.49; 95% CONFIDENCE INTERVAL 1.12-1.97; P = 0.006) AND WAS ASSOCIATED WITH ER-ALPHA BIOACTIVITY IN THE CORRESPONDING SERUM. CONCLUSION/SIGNIFICANCE: THIS FIRST LARGE-SCALE EPIGENOTYPING STUDY DEMONSTRATES THAT DNA METHYLATION MAY SERVE AS A LINK BETWEEN THE ENVIRONMENT AND THE GENOME. FACTORS THAT CAN BE MODULATED BY THE ENVIRONMENT (LIKE ESTROGENS) LEAVE AN IMPRINT IN THE DNA OF CELLS THAT ARE UNRELATED TO THE TARGET ORGAN AND INDICATE THE PREDISPOSITION TO DEVELOP A CANCER. FURTHER RESEARCH WILL NEED TO DEMONSTRATE WHETHER DNA METHYLATION PROFILES WILL BE ABLE TO SERVE AS A NEW TOOL TO PREDICT THE RISK OF DEVELOPING CHRONIC DISEASES WITH SUFFICIENT ACCURACY TO GUIDE PREVENTIVE MEASURES.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
14 6488  38 TP53 R72P POLYMORPHISM MODULATES DNA METHYLATION IN HEPATOCELLULAR CARCINOMA. BACKGROUND: HEPATOCELLULAR CARCINOMA (HCC) IS CHARACTERIZED BY WIDESPREAD EPIDEMIOLOGICAL AND MOLECULAR HETEROGENEITY. PREVIOUS WORK SHOWED THAT IN THE WESTERN PART OF NORTH AFRICA, A REGION OF LOW INCIDENCE OF HCC, MUTATIONS ARE SCARCE FOR THIS TUMOR TYPE. AS EPIGENETIC CHANGES ARE CONSIDERED POSSIBLE SURROGATES TO MUTATIONS IN HUMAN CANCERS, WE DECIDED, THUS, TO CHARACTERIZE DNA METHYLATION IN HCC FROM NORTH-AFRICAN PATIENTS. METHODS: A SET OF 11 LOCI WAS INVESTIGATED IN A SERIES OF 45 TUMOR SPECIMENS USING METHYLATION-SPECIFIC AND COMBINED-BISULFITE RESTRICTION ASSAY PCR. RESULTS OBTAINED ON CLINICAL SAMPLES WERE SUBSEQUENTLY VALIDATED IN LIVER CANCER CELL LINES. RESULTS: DNA METHYLATION AT TUMOR SUPPRESSOR LOCI IS SIGNIFICANTLY HIGHER IN SAMPLES DISPLAYING CHROMOSOME INSTABILITY. MORE IMPORTANTLY, DNA METHYLATION WAS SIGNIFICANTLY HIGHER IN ARG/ARG WHEN COMPARED TO PRO/PRO GENOTYPE CARRIERS AT CODON 72 RS1042522 OF TP53 (65% VS 20% METHYLATED LOCI, P = 0.0006), A POLYMORPHISM ALREADY KNOWN TO AFFECT SOMATIC MUTATION RATE IN HUMAN CARCINOMAS. IN VITRO EXPERIMENTS IN CELL LINES INDICATED THAT ENZYMES CONTROLLING DNA METHYLATION WERE DIFFERENTIALLY REGULATED BY CODON 72 ARG OR PRO ISOFORMS OF P53. FURTHERMORE, THE ARG72-CARRYING VERSION OF P53 WAS SHOWN TO RE-METHYLATE DNA MORE RAPIDLY THAN THE PRO-HARBORING ISOFORM. FINALLY, PRO-CARRYING CELL LINES WERE SHOWN TO BE SIGNIFICANTLY MORE RESISTANT TO DECITABINE TREATMENT (TWO-FOLD, P = 0.005). CONCLUSIONS: OUR DATA SUGGEST THAT ARG72PRO POLYMORPHISM IN A WT P53 CONTEXT MAY ACT AS A PRIMARY DRIVER OF EPIGENETIC CHANGES IN HCC. IT SUGGESTS, IN ADDITION, THAT RS1042522 GENOTYPE MAY PREDICT SENSITIVITY TO EPIGENETIC-TARGETED THERAPY. THIS MODEL OF LIVER TUMORIGENESIS THAT ASSOCIATES LOW PENETRANCE GENETIC PREDISPOSITION TO EPIGENETIC CHANGES EMERGES FROM A REGION OF LOW HCC INCIDENCE AND IT MAY, THEREFORE, APPLY ESSENTIALLY TO POPULATION LIVING IN SIMILAR AREAS. SURVEYS ON POPULATIONS SUBMITTED TO HIGHLY MUTAGENIC CONDITIONS AS PERINATALLY-ACQUIRED CHRONIC HEPATITIS B OR AFLATOXIN B1 EXPOSURE REMAINED TO BE CONDUCTED TO VALIDATE OUR OBSERVATIONS AS A GENERAL MODEL.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
15 4359  39 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
16 1117  30 COMPARATIVE AND EXPERIMENTAL STUDIES ON THE GENES ALTERED BY CHRONIC HYPOXIA IN HUMAN BRAIN MICROENDOTHELIAL CELLS. BACKGROUND : HYPOXIA INDUCIBLE FACTOR 1 ALPHA (HIF1A) IS A MASTER REGULATOR OF ACUTE HYPOXIA; HOWEVER, WITH CHRONIC HYPOXIA, HIF1A LEVELS RETURN TO THE NORMOXIC LEVELS. IMPORTANTLY, THE GENES THAT ARE INVOLVED IN THE CELL SURVIVAL AND VIABILITY UNDER CHRONIC HYPOXIA ARE NOT KNOWN. THEREFORE, WE TESTED THE HYPOTHESIS THAT CHRONIC HYPOXIA LEADS TO THE UPREGULATION OF A CORE GROUP OF GENES WITH ASSOCIATED CHANGES IN THE PROMOTER DNA METHYLATION THAT MEDIATES THE CELL SURVIVAL UNDER HYPOXIA. RESULTS : WE EXAMINED THE EFFECT OF CHRONIC HYPOXIA (3 DAYS; 0.5% OXYGEN) ON HUMAN BRAIN MICRO ENDOTHELIAL CELLS (HBMEC) VIABILITY AND APOPTOSIS. HYPOXIA CAUSED A SIGNIFICANT REDUCTION IN CELL VIABILITY AND AN INCREASE IN APOPTOSIS. NEXT, WE EXAMINED CHRONIC HYPOXIA ASSOCIATED CHANGES IN TRANSCRIPTOME AND GENOME-WIDE PROMOTER METHYLATION. THE DATA OBTAINED WAS COMPARED WITH 16 OTHER MICROARRAY STUDIES ON CHRONIC HYPOXIA. NINE GENES WERE ALTERED IN RESPONSE TO CHRONIC HYPOXIA IN ALL 17 STUDIES. INTERESTINGLY, HIF1A WAS NOT ALTERED WITH CHRONIC HYPOXIA IN ANY OF THE STUDIES. FURTHERMORE, WE COMPARED OUR DATA TO THREE OTHER STUDIES THAT IDENTIFIED HIF-RESPONSIVE GENES BY VARIOUS APPROACHES. ONLY TWO GENES WERE FOUND TO BE HIF DEPENDENT. WE SILENCED EACH OF THESE 9 GENES USING CRISPR/CAS9 SYSTEM. DOWNREGULATION OF EGLN3 SIGNIFICANTLY INCREASED THE CELL DEATH UNDER CHRONIC HYPOXIA, WHEREAS DOWNREGULATION OF ERO1L, ENO2, ADRENOMEDULLIN, AND SPAG4 REDUCED THE CELL DEATH UNDER HYPOXIA. CONCLUSIONS : WE PROVIDE A CORE GROUP OF GENES THAT REGULATES CELLULAR ACCLIMATIZATION UNDER CHRONIC HYPOXIC STRESS, AND MOST OF THEM ARE HIF INDEPENDENT.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
17 5785  30 SPONTANEOUS NEOPLASTIC TRANSFORMATION OF WB-F344 RAT LIVER EPITHELIAL CELLS. SEVERAL STUDIES HAVE SHOWN THAT CULTURED RAT LIVER EPITHELIAL CELLS TRANSFORM SPONTANEOUSLY AFTER CHRONIC MAINTENANCE IN A CONFLUENT STATE IN VITRO. IN THE PRESENT STUDY, MULTIPLE INDEPENDENT LINEAGES OF LOW-PASSAGE WB-F344 RAT LIVER EPITHELIAL STEM-LIKE CELLS WERE INITIATED AND SUBJECTED IN PARALLEL TO SELECTION FOR SPONTANEOUS TRANSFORMATION TO DETERMINE WHETHER SPONTANEOUS ACQUISITION OF TUMORIGENICITY WAS THE RESULT OF EVENTS (GENETIC OR EPIGENETIC) THAT OCCURRED INDEPENDENTLY AND STOCHASTICALLY, OR REFLECTED THE EXPRESSION OF A PRE-EXISTING ALTERATION WITHIN THE PARENTAL WB-F344 CELL LINE. TEMPORAL ANALYSIS OF THE SPONTANEOUS ACQUISITION OF TUMORIGENICITY BY WB-F344 CELLS DEMONSTRATED LINEAGE-SPECIFIC DIFFERENCES IN THE TIME OF FIRST EXPRESSION OF THE TUMORIGENIC PHENOTYPE, FREQUENCIES AND LATENCIES OF TUMOR FORMATION, AND TUMOR DIFFERENTIATIONS. ALTHOUGH SPONTANEOUSLY TRANSFORMED WB-F344 CELLS PRODUCED DIVERSE TUMOR TYPES (INCLUDING HEPATOCELLULAR CARCINOMAS, CHOLANGIOCARCINOMAS, HEPATOBLASTOMAS, AND OSTEOGENIC SARCOMAS), INDIVIDUAL LINEAGES YIELDED TUMORS WITH CONSISTENT AND SPECIFIC PATTERNS OF DIFFERENTIATION. THESE RESULTS PROVIDE SUBSTANTIAL EVIDENCE THAT THE STOCHASTIC ACCUMULATION OF INDEPENDENT TRANSFORMING EVENTS DURING THE SELECTION REGIMEN IN VITRO WERE RESPONSIBLE FOR SPONTANEOUS NEOPLASTIC TRANSFORMATION OF WB-F344 CELLS. FURTHERMORE, CELL LINEAGE COMMITMENT TO A SPECIFIC DIFFERENTIATION PROGRAM WAS STABLE WITH TIME IN CULTURE AND WITH SITE OF TRANSPLANTATION. THIS IS THE FIRST REPORT OF A COHORT OF RELATED, BUT INDEPENDENT, RAT LIVER EPITHELIAL CELL LINES THAT COLLECTIVELY PRODUCE A SPECTRUM OF TUMOR TYPES BUT INDIVIDUALLY REPRODUCE A SPECIFIC TUMOR TYPE. THESE CELL LINES WILL PROVIDE VALUABLE REAGENTS FOR INVESTIGATION OF THE MOLECULAR MECHANISMS INVOLVED IN THE DIFFERENTIATION OF HEPATIC STEM-LIKE CELLS AND FOR EXAMINATION OF POTENTIAL CAUSAL RELATIONSHIPS IN SPONTANEOUSLY TRANSFORMED RAT LIVER EPITHELIAL CELL LINES BETWEEN MOLECULAR/CELLULAR ALTERATIONS AND THE ABILITY TO PRODUCE TUMORS IN SYNGENEIC ANIMALS.	1998	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
18 5614  42 SAFETY AND EFFICACY OF EPIGENETICALLY CONVERTED HUMAN FIBROBLASTS INTO INSULIN-SECRETING CELLS: A PRECLINICAL STUDY. TYPE 1 DIABETES MELLITUS (T1DM) IS A CHRONIC DISEASE THAT LEADS TO LOSS OF INSULIN SECRETING BETA-CELLS, CAUSING HIGH LEVELS OF BLOOD GLUCOSE. EXOGENOUS INSULIN ADMINISTRATION IS NOT SUFFICIENT TO MIMIC THE NORMAL FUNCTION OF BETA-CELLS AND, CONSEQUENTLY, DIABETES MELLITUS OFTEN PROGRESSES AND CAN LEAD TO MAJOR CHRONIC COMPLICATIONS AND MORBIDITY. THE PHYSIOLOGICAL CONTROL OF GLUCOSE LEVELS CAN ONLY BE RESTORED BY REPLACING THE BETA-CELL MASS.WE RECENTLY DEVELOPED A NEW STRATEGY THAT ALLOWS FOR EPIGENETIC CONVERSION OF DERMAL FIBROBLASTS INTO INSULIN-SECRETING CELLS (EPICC), USING A BRIEF EXPOSURE TO THE DEMETHYLATING AGENT 5-AZA-CYTIDINE (5-AZA-CR), FOLLOWED BY A PANCREATIC INDUCTION PROTOCOL. THIS METHOD HAS NOTABLE ADVANTAGES COMPARED TO THE ALTERNATIVE AVAILABLE PROCEDURES AND MAY REPRESENT A PROMISING TOOL FOR CLINICAL TRANSLATION AS A THERAPY FOR T1DM. HOWEVER, A THOUGHT EVALUATION OF ITS THERAPEUTIC SAFETY AND EFFICACY IS MANDATORY TO SUPPORT PRECLINICAL STUDIES BASED ON EPICC TREATMENT.WE HERE REPORT THE DATA OBTAINED USING HUMAN FIBROBLASTS ISOLATED FROM DIABETIC AND HEALTHY INDIVIDUALS, BELONGING THE TWO GENDERS. EPICC WERE INJECTED INTO 650 DIABETIC SEVERE COMBINED IMMUNODEFICIENCY (SCID) MICE AND DEMONSTRATED TO BE ABLE TO RESTORE AND MAINTAIN GLYCEMIC LEVELS WITHIN THE PHYSIOLOGICAL RANGE. CELLS HAD THE ABILITY TO SELF-REGULATE AND NOT TO CAUSE HYPOGLYCEMIA, WHEN TRANSPLANTED IN HEALTHY ANIMALS. EFFICACY TESTS SHOWED THAT EPICC SUCCESSFULLY RE-ESTABLISHED NORMOGLYCEMIA IN DIABETIC MICE, USING A DOSE RANGE THAT APPEARED CLINICALLY RELEVANT TO THE CONCENTRATION 0.6 X 10(6) EPICC. NECROPSY AND HISTOPATHOLOGICAL INVESTIGATIONS DEMONSTRATED THE ABSENCE OF MALIGNANT TRANSFORMATION AND CELL MIGRATION TO ORGANS AND LYMPH NODES.THE PRESENT PRECLINICAL STUDY DEMONSTRATES SAFETY AND EFFICACY OF HUMAN EPICC IN DIABETIC MICE AND SUPPORTS THE USE OF EPIGENETIC CONVERTED CELLS FOR REGENERATIVE MEDICINE OF DIABETES MELLITUS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
19 2297  27 EPIGENETIC REGULATION OF ACUTE INFLAMMATORY PAIN. ACUTE PAIN IS ASSOCIATED WITH TISSUE DAMAGE, WHICH RESULTS IN THE RELEASE OF INFLAMMATORY MEDIATORS. RECENT STUDIES POINT TO THE INVOLVEMENT OF EPIGENETIC MECHANISMS (DNA METHYLATION) IN THE DEVELOPMENT OF PAIN. WE HAVE FOUND THAT DURING ACUTE INFLAMMATORY PAIN INDUCED BY THE APPLICATION OF 10% MUSTARD OIL ON THE TONGUES OF RATS, LEVELS OF DNMT3A AND 3B WERE ELEVATED MARKEDLY (36 AND 42 % RESPECTIVELY), WHEREAS THE LEVEL OF DNMT1 WAS NOT CHANGED SIGNIFICANTLY. PREVIOUS INJECTION OF XEFOCAM WITH 0,4 MG/KG DOSE DECREASED LEVELS OF DNMT3A AND 3B (25 AND 24% RESPECTIVELY). THE LEVEL OF DNMT1 WAS NOT CHANGED SIGNIFICANTLY COMPARED TO THE CONTROL GROUP. THE FINDINGS SUPPORT THE IDEA THAT INHIBITORS OF DNA-METHYLTRANSFERASES COULD BE USEFUL FOR PAIN MANAGEMENT. OUR DATA SUGGEST THAT NSAIDS (ALONE OR IN COMBINATION WITH DNMT INHIBITORS) MAY BE PROPOSED AS POSSIBLE EPIGENETIC REGULATORY AGENTS, WHICH MAY PLAY A ROLE IN EPIGENETIC MECHANISMS INDIRECTLY THROUGH ALTERING THE ACTIVITY OF INFLAMMATORY MEDIATORS INVOLVED IN PAIN DEVELOPMENT.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
20 1567  33 DNA METHYLATION OF THE KLF14 GENE REGION IN WHOLE BLOOD CELLS PROVIDES PREDICTION FOR THE CHRONIC INFLAMMATION IN THE ADIPOSE TISSUE. KRUPPEL-LIKE FACTOR 14 (KLF14) GENE, WHICH APPEARS TO BE A MASTER REGULATOR OF GENE EXPRESSION IN THE ADIPOSE TISSUE AND HAVE PREVIOUSLY BEEN ASSOCIATED WITH BMI AND TYPE 2 DIABETES (T2D) BY LARGE GENOME-WIDE ASSOCIATION STUDIES. IN ORDER TO FIND PREDICTIVE BIOMARKERS FOR THE DEVELOPMENT OF T2D, IT IS NECESSARY TO TAKE EPIGENOMIC CHANGES AFFECTED BY ENVIRONMENTAL FACTORS INTO ACCOUNT. THIS STUDY FOCUSES ON AGEING AND OBESITY, WHICH ARE T2D RISK FACTORS, AND EXAMINES EPIGENETIC CHANGES AND INFLAMMATORY CHANGES. WE INVESTIGATED DNA METHYLATION CHANGES IN THE KLF14 PROMOTER REGION IN DIFFERENT ORGANS OF MICE FOR COMPARING AGING AND WEIGHT. WE FOUND THAT METHYLATION LEVELS OF THESE SITES WERE INCREASED WITH AGING AND WEIGHT IN THE SPLEEN, THE ADIPOSE TISSUE, THE KIDNEY, THE LUNG, THE COLON AND THE WHOLE BLOOD CELLS. IN ADDITION, IN THE SPLEEN, THE ADIPOSE TISSUE AND THE WHOLE BLOOD, THESE EPIGENETIC CHANGES WERE ALSO SIGNIFICANTLY ASSOCIATED WITH INFLAMMATORY LEVELS. MOREOVER, NOT ONLY KLF14, BUT ALSO EXPRESSION LEVELS OF SOME DOWNSTREAM GENES WERE DECREASED WITH METHYLATION IN THE SPLEEN, THE ADIPOSE TISSUE AND THE WHOLE BLOOD CELLS. TAKEN TOGETHER, OUR RESULTS SUGGEST THAT METHYLATION CHANGES OF KLF14 IN THOSE TISSUES MAY BE ASSOCIATED WITH CHANGES IN GENE EXPRESSION AND INFLAMMATION ON THE ADIPOSE TISSUE OF OBESITY AND T2D. IN ADDITION, THE METHYLATION CHANGES IN THE WHOLE BLOOD CELLS MAY SERVE AS A PREDICTIVE EPIGENETIC BIOMARKER FOR THE DEVELOPMENT OF T2D.	2018