1 6514 126 TRANSCRIPTIONAL ACTIVATION OF THE GP91PHOX NADPH OXIDASE SUBUNIT BY TPA IN HL-60 CELLS. THE EXPOSURE TO EPIGENETIC EFFECTORS CAPABLE OF INDUCING COPIOUS PRODUCTION OF REACTIVE OXYGEN SPECIES (ROS) HAS BEEN ASSOCIATED WITH CHRONIC INFLAMMATION, TUMOR INITIATION, AND PROMOTION. THE OBJECTIVE OF THIS STUDY WAS TO EXAMINE THE REGULATION OF GP91PHOX, THE CATALYTIC SUBUNIT OF THE NADPH OXIDASE, AND THE KINETICS OF ROS PRODUCTION IN PROMYELOCYTIC LEUKEMIA HL-60 CELLS INDUCED WITH 12-O-TETRADECONYLPHORBOL-13-ACETATE (TPA). THE TREATMENT OF HL-60 CELLS WITH TPA (0.1 MICROM) INDUCED CELLULAR DIFFERENTIATION, WHICH WAS FOLLOWED AFTER 48 H BY A TENFOLD INCREASE IN CHEMILUMINESCENCE FROM LUCIGENIN AND A 2.5-FOLD INCREASE IN THE INTRACELLULAR OXIDATION OF 2',7'-DICHOLOROFLUORESCIN (DCFH). WHEREAS HIGHER CONCENTRATIONS (1.0 MICROM) OF TPA DID NOT STIMULATE FURTHER ROS PRODUCTION, REPEATED STIMULATION WITH 0.1 MICROM TPA OF DIFFERENTIATED CELLS INDUCED A MODEST (1.2-FOLD) BUT RAPID (15 MIN) INCREASE IN CHEMILUMINESCENCE. IN CELLS TREATED WITH TPA, THE BURST IN ROS AT 48 H WAS PRECEDED BY ACCUMULATION AT 12 H OF GP91PHOX (8.8-FOLD) AND P47PHOX MRNA (THREEFOLD), WHEREAS UNTREATED CELLS CONTAINED STEADY-STATE LEVELS OF BOTH TRANSCRIPTS. TIME-COURSE EXPERIMENTS WITH ACTINOMYCIN D TO INHIBIT TRANSCRIPTION REVEALED THAT TPA DID NOT IMPROVE THE STABILITY OF GP91PHOX. IN TRANSIENT TRANSFECTIONS, LUCIFERASE REPORTER ACTIVITY DIRECTED FROM A 1.5-KB GP91PHOX PROMOTER FRAGMENT WAS ENHANCED THREEFOLD UPON TREATMENT WITH TPA FOR 24 H. WE CONCLUDE THAT TPA CAN COMMIT HL-60 CELLS TO DIFFERENTIATION AND ELICIT TRANSCRIPTION FROM THE PROXIMAL GP91PHOX PROMOTER. 2001 2 1426 25 DIFFERENTIAL EFFECTS OF BUTYRIC ACID ON MOUSE SKIN TUMORIGENESIS. WE STUDIED THE EFFECTS OF BUTYRIC ACID (BA) ON MOUSE SKIN TUMORIGENESIS USING CHRONIC ANIMAL BIOASSAYS. TOPICAL APPLICATION OF BA IMMEDIATELY AFTER EACH TREATMENT WITH 12-0-TETRADECANOYLPHORBOL-13-ACETATE (TPA) PROMOTER-INHIBITED SKIN TUMORS. THE EFFECT WAS DEPENDENT ON THE DOSE OF BA APPLIED. BA SHOWED NO MARKED INHIBITORY EFFECT ON EITHER SKIN TUMOR INITIATION OR COMPLETE TUMORIGENESIS INDUCED BY DIMETHYLBENZANTHRACENE (DMBA). SINCE TUMOR PROMOTION REPORTEDLY INVOLVES EPIGENETIC EVENTS WHEREAS TUMOR INITIATION OR COMPLETE TUMORIGENESIS TAKES PLACE THROUGH GENETIC PATHWAYS, IT IS POSSIBLE THAT BA EXERTS ITS ANTITUMORIGENIC EFFECTS MAINLY BY ALTERING THE EPIGENETIC EVENTS RESPONSIBLE FOR TUMOR PROMOTION. THE RESULTS OF THE STUDY COULD FURTHER BE USED TO STUDY THE MECHANISM OF ACTION AND MODIFICATION OF ANTITUMORIGENIC EFFECTS OF BA IN COMBINATION WITH OTHER SUBSTANCES. 1997 3 2887 22 GADD45A TRANSCRIPTIONAL INDUCTION ELICITED BY THE AURORA KINASE INHIBITOR MK-0457 IN BCR-ABL-EXPRESSING CELLS IS DRIVEN BY OCT-1 TRANSCRIPTION FACTOR. THE ADVANTAGE OF AURORA KINASE (AK) INHIBITORS IN CHRONIC MYELOID LEUKEMIA (CML) THERAPY MOSTLY ARISES FROM "OFF-TARGET" EFFECTS ON TYROSINE KINASE (TK) ACTIVITY OF WILD TYPE (WT) OR MUTATED BCR-ABL PROTEINS WHICH DRIVE THE DISEASE RESISTANCE TO IMATINIB (IM). WE PROVED THAT THE AK INHIBITOR MK-0457 INDUCES THE GROWTH ARREST DNA DAMAGE-INDUCIBLE (GADD) 45A THROUGH RECRUITMENT OF OCTAMER-BINDING (OCT)-1 TRANSCRIPTION FACTOR AT A CRITICAL PROMOTER REGION FOR GENE TRANSCRIPTION AND COVALENT MODIFICATIONS OF HISTONE H3 (LYSINE 14 ACETYLATION, LYSINE 9 DE-METHYLATION). SUCH EPIGENETIC CHROMATIN MODIFICATIONS MAY DEPICT A GENERAL MECHANISM PROMOTING THE RE-ACTIVATION OF TUMOR SUPPRESSOR GENES SILENCED BY BCR-ABL. 2012 4 4048 28 MAINTENANCE AND PHARMACOLOGIC TARGETING OF ROR1 PROTEIN LEVELS VIA UHRF1 IN T(1;19) PRE-B-ALL. EXPRESSION OF THE TRANSMEMBRANE PSEUDOKINASE ROR1 IS REQUIRED FOR SURVIVAL OF T(1;19)-PRE-B-CELL ACUTE LYMPHOBLASTIC LEUKEMIA (T(1;19) PRE-B-ALL), CHRONIC LYMPHOCYTIC LEUKEMIA, AND MANY SOLID TUMORS. HOWEVER, TARGETING ROR1 WITH SMALL-MOLECULES HAS BEEN CHALLENGING DUE TO THE ABSENCE OF ROR1 KINASE ACTIVITY. TO IDENTIFY GENES THAT REGULATE ROR1 EXPRESSION AND MAY, THEREFORE, SERVE AS SURROGATE DRUG TARGETS, WE EMPLOYED AN SIRNA SCREENING APPROACH AND DETERMINED THAT THE EPIGENETIC REGULATOR AND E3 UBIQUITIN LIGASE, UHRF1, IS REQUIRED FOR T(1;19) PRE-B-ALL CELL VIABILITY IN A ROR1-DEPENDENT MANNER. UPON UHRF1 SILENCING, ROR1 PROTEIN IS REDUCED WITHOUT ALTERING ROR1 MRNA, AND ECTOPICALLY EXPRESSED UHRF1 IS SUFFICIENT TO INCREASE ROR1 LEVELS. ADDITIONALLY, PROTEASOME INHIBITION RESCUES LOSS OF ROR1 PROTEIN AFTER UHRF1 SILENCING, SUGGESTING A ROLE FOR THE PROTEASOME IN THE UHRF1-ROR1 AXIS. FINALLY, WE SHOW THAT ROR1-POSITIVE CELLS ARE TWICE AS SENSITIVE TO THE UHRF1-TARGETING DRUG, NAPHTHAZARIN, AND UNDERGO INCREASED APOPTOSIS COMPARED TO ROR1-NEGATIVE CELLS. NAPHTHAZARIN ELICITS REDUCED EXPRESSION OF UHRF1 AND ROR1, AND COMBINATION OF NAPHTHAZARIN WITH INHIBITORS OF PRE-B CELL RECEPTOR SIGNALING RESULTS IN FURTHER REDUCTION OF CELL SURVIVAL COMPARED WITH EITHER INHIBITOR ALONE. THEREFORE, OUR WORK REVEALS A MECHANISM BY WHICH UHRF1 STABILIZES ROR1, SUGGESTING A POTENTIAL TARGETING STRATEGY TO INHIBIT ROR1 IN T(1;19) PRE-B-ALL AND OTHER MALIGNANCIES. 2018 5 3620 20 IN VIVO AND IN VITRO GENOTOXIC AND EPIGENETIC EFFECTS OF TWO TYPES OF COLA BEVERAGES AND CAFFEINE: A MULTIASSAY APPROACH. THE AIM OF THIS WORK WAS TO ASSESS THE BIOLOGICAL AND FOOD SAFETY OF TWO DIFFERENT BEVERAGES: CLASSIC COCA COLA (CCC) AND CAFFEINE-FREE COCA COLA (CFCC). TO THIS END, WE DETERMINED THE GENOTOXICOLOGICAL AND BIOLOGICAL EFFECTS OF DIFFERENT DOSES OF LYOPHILISED CCC AND CFCC AND CAFFEINE (CAF), THE MAIN DISTINCTIVE CONSTITUENT. THEIR TOXIC/ANTITOXIC, GENOTOXIC/ANTIGENOTOXIC, AND CHRONIC TOXICITY (LIFESPAN ASSAY) EFFECTS WERE DETERMINED IN VIVO USING THE DROSOPHILA MODEL. THEIR CYTOTOXIC ACTIVITIES WERE DETERMINED USING THE HL-60 IN VITRO CANCER MODEL. IN ADDITION, CLASTOGENIC DNA TOXICITY WAS MEASURED USING INTERNUCLEOSOMAL FRAGMENTATION AND SCGE ASSAYS. THEIR EPIGENETIC EFFECTS WERE ASSESSED ON THE HL-60 METHYLATION STATUS USING SOME REPETITIVE ELEMENTS. THE EXPERIMENTAL RESULTS SHOWED A SLIGHT CHEMOPREVENTIVE EFFECT OF THE TWO COLA BEVERAGES AGAINST HL-60 LEUKAEMIA CELLS, PROBABLY MEDIATED BY NONAPOPTOTIC MECHANISMS. FINALLY, CCC AND CAF INDUCED A GLOBAL GENOME HYPOMETHYLATION EVALUATED IN LINE-1 AND ALU M1 REPETITIVE ELEMENTS. OVERALL, WE DEMONSTRATED FOR THE FIRST TIME THE SAFETY OF THIS FAMOUS BEVERAGE IN IN VIVO AND IN VITRO MODELS. 2016 6 967 30 CHRONIC NICOTINE EXPOSURE AUGMENTS RENAL OXIDATIVE STRESS AND INJURY THROUGH TRANSCRIPTIONAL ACTIVATION OF P66SHC. BACKGROUND: CHRONIC NICOTINE (CH-NIC) EXPOSURE EXACERBATES ISCHEMIA/REPERFUSION (I/R)-INDUCED OXIDATIVE STRESS AND ACUTE KIDNEY INJURY (AKI), AND MITOCHONDRIAL PRODUCTION OF REACTIVE OXYGEN SPECIES (ROS) IN CULTURED RENAL PROXIMAL TUBULE CELLS (RPTCS). BECAUSE SER36-PHOSPHORYLATED P66SHC MODULATES MITOCHONDRIAL ROS PRODUCTION AND INJURY OF RPTCS, WE HYPOTHESIZED THAT CH-NIC EXACERBATES AKI BY INCREASING STRESS-INDUCED PHOSPHORYLATION OF P66SHC. METHODS: WE FIRST TESTED WHETHER CH-NIC AUGMENTS I/R-AKI-INDUCED EXPRESSION AND PHOSPHORYLATION OF P66SHC IN VIVO. WE THEN EXAMINED WHETHER KNOCKING DOWN P66SHC, OR IMPAIRING ITS SER36 PHOSPHORYLATION OR BINDING TO CYTOCHROME C, ALTERS THE EFFECTS OF CH-NIC ON OXIDATIVE STRESS (H(2)O(2))-INDUCED PRODUCTION OF ROS, MITOCHONDRIAL DEPOLARIZATION AND INJURY IN RPTCS IN VITRO. RESULTS: WE FOUND THAT CH-NIC INCREASED THE EXPRESSION OF P66SHC IN THE CONTROL AND ISCHEMIC KIDNEYS, BUT ONLY INCREASED ITS SER36 PHOSPHORYLATION AFTER RENAL I/R. KNOCKING DOWN P66SHC OR IMPAIRING PHOSPHORYLATION OF ITS SER36 RESIDUE, VIA THE S36A MUTATION (BUT NOT THE PHOSPHOMIMETIC S36D MUTATION), BLUNTED CH-NIC + H2O2-DEPENDENT ROS PRODUCTION, MITOCHONDRIAL DEPOLARIZATION AND INJURY IN RPTCS. ADDITIONALLY, CH-NIC + H2O2-DEPENDENT BINDING OF P66SHC TO MITOCHONDRIAL CYTOCHROME C WAS ATTENUATED BY S36A MUTATION OF P66SHC, AND IMPAIRING CYTOCHROME C BINDING (VIA W134F MUTATION) ABOLISHED ROS PRODUCTION, MITOCHONDRIAL DEPOLARIZATION AND INJURY, WHILE ECTOPIC OVEREXPRESSION OF P66SHC (WHICH MIMICS CH-NIC TREATMENT) AUGMENTED OXIDANT INJURY. WE DETERMINED THAT CH-NIC STIMULATES THE P66SHC PROMOTER THROUGH P53- AND EPIGENETIC MODIFICATION (PROMOTER HYPOMETHYLATION). CONCLUSIONS: CH-NIC WORSENS OXIDATIVE STRESS-DEPENDENT ACUTE RENAL INJURY BY INCREASING EXPRESSION AND CONSEQUENT OXIDATIVE STRESS-DEPENDENT SER36 PHOSPHORYLATION OF P66SHC. THUS, TARGETING THIS PATHWAY MAY HAVE THERAPEUTIC RELEVANCE IN PREVENTING/AMELIORATING TOBACCO-RELATED KIDNEY INJURY. 2013 7 671 30 BOTHROPS MOOJENI L-AMINO ACID OXIDASE INDUCES APOPTOSIS AND EPIGENETIC MODULATION ON BCR-ABL(+) CELLS. BACKGROUND: RESISTANCE TO APOPTOSIS IN CHRONIC MYELOID LEUKEMIA (CML) IS ASSOCIATED WITH CONSTITUTIVE TYROSINE KINASE ACTIVITY OF THE BCR-ABL ONCOPROTEIN. THE DEREGULATED EXPRESSION OF APOPTOSIS-RELATED GENES AND ALTERATION IN EPIGENETIC MACHINERY MAY ALSO CONTRIBUTE TO APOPTOSIS RESISTANCE IN CML. TYROSINE KINASE INHIBITORS TARGET THE BCR-ABL ONCOPROTEIN AND ARE USED IN CML TREATMENT. THE RESISTANCE OF CML PATIENTS TO TYROSINE KINASE INHIBITORS HAS GUIDED THE SEARCH FOR NEW COMPOUNDS THAT MAY INDUCE APOPTOSIS IN BCR-ABL(+) LEUKEMIC CELLS AND IMPROVE THE DISEASE TREATMENT. METHODS: IN THE PRESENT STUDY, WE INVESTIGATED WHETHER THE L-AMINO ACID OXIDASE ISOLATED FROM BOTHROPS MOOJENI SNAKE VENOM (BMOOLAAO-I) (I) WAS CYTOTOXIC TO BCR-ABL(+) CELL LINES (HL-60.BCR-ABL, K562-S, AND K562-R), HL-60 (ACUTE PROMYELOCYTIC LEUKEMIA) CELLS, THE NON-TUMOR CELL LINE HEK-293, AND PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMC); AND (II) AFFECTED EPIGENETIC MECHANISMS, INCLUDING DNA METHYLATION AND MICRORNAS EXPRESSION IN VITRO. RESULTS: BMOOLAAO-I INDUCED ROS PRODUCTION, APOPTOSIS, AND DIFFERENTIAL DNA METHYLATION PATTERN OF REGULATORY APOPTOSIS GENES. THE TOXIN UPREGULATED EXPRESSION OF THE PRO-APOPTOTIC GENES BID AND FADD AND DOWNREGULATED DFFA EXPRESSION IN LEUKEMIC CELL LINES, AS WELL AS INCREASED MIR-16 EXPRESSION - WHOSE MAJOR PREDICTED TARGET IS THE ANTI-APOPTOTIC GENE BCL2 - IN BCR-ABL(+) CELLS. CONCLUSION: BMOOLAAO-I EXERTS SELECTIVE ANTITUMOR ACTION MEDIATED BY H(2)O(2) RELEASE AND INDUCES APOPTOSIS, AND ALTERATIONS IN EPIGENETIC MECHANISMS. THESE RESULTS SUPPORT FUTURE INVESTIGATIONS ON THE EFFECT OF BMOOLAAO-I ON IN VIVO MODELS TO DETERMINE ITS POTENTIAL IN CML THERAPY. 2020 8 2311 27 EPIGENETIC REGULATION OF CYTOSOLIC PHOSPHOLIPASE A2 IN SH-SY5Y HUMAN NEUROBLASTOMA CELLS. GROUP IVA CYTOSOLIC PHOSPHOLIPASE A2 (CPLA2 OR PLA2G4A) IS A KEY ENZYME THAT CONTRIBUTES TO INFLAMMATION VIA THE GENERATION OF ARACHIDONIC ACID AND EICOSANOIDS. WHILE MUCH IS KNOWN ABOUT REGULATION OF CPLA2 BY POSTTRANSLATIONAL MODIFICATION SUCH AS PHOSPHORYLATION, LITTLE IS KNOWN ABOUT ITS EPIGENETIC REGULATION. IN THIS STUDY, TREATMENT WITH HISTONE DEACETYLASE (HDAC) INHIBITORS, TRICHOSTATIN A (TSA), VALPROIC ACID, TUBACIN AND THE CLASS I HDAC INHIBITOR, MS-275, WERE FOUND TO INCREASE CPLA2ALPHA MESSENGER RNA (MRNA) EXPRESSION IN SH-SY5Y HUMAN NEUROBLASTOMA CELLS. CO-TREATMENT OF THE HISTONE ACETYLTRANSFERASE (HAT) INHIBITOR, ANACARDIC ACID, MODULATED UPREGULATION OF CPLA2ALPHA INDUCED BY TSA. SPECIFIC INVOLVEMENT OF CLASS I HDACS AND HAT IN CPLA2ALPHA REGULATION WAS FURTHER SHOWN, AND A TIP60-SPECIFIC HAT INHIBITOR, NU9056, MODULATED THE UPREGULATION OF CPLA2ALPHA INDUCED BY MS-275. IN ADDITION, CO-TREATMENT OF WITH HISTONE METHYLTRANSFERASE (HMT) INHIBITOR, 5'-DEOXY-5'-METHYLTHIOADENOSINE (MTA) SUPPRESSED TSA-INDUCED CPLA2ALPHA UPREGULATION. THE ABOVE CHANGES IN CPLA2 MRNA EXPRESSION WERE REFLECTED AT THE PROTEIN LEVEL BY WESTERN BLOTS AND IMMUNOCYTOCHEMISTRY. CHROMATIN IMMUNOPRECIPITATION (CHIP) SHOWED TSA INCREASED BINDING OF TRIMETHYLATED H3K4 TO THE PROXIMAL PROMOTER REGION OF THE CPLA2ALPHA GENE. CELL INJURY AFTER TSA TREATMENT AS INDICATED BY LACTATE DEHYDROGENASE (LDH) RELEASE WAS MODULATED BY ANACARDIC ACID, AND A ROLE OF CPLA2 IN MEDIATING TSA-INDUCED INJURY SHOWN, AFTER CO-INCUBATION WITH THE CPLA2 SELECTIVE INHIBITOR, ARACHIDONOYL TRIFLUOROMETHYL KETONE (AACOCF3). TOGETHER, RESULTS INDICATE EPIGENETIC REGULATION OF CPLA2 AND THE POTENTIAL OF SUCH REGULATION FOR TREATMENT OF CHRONIC INFLAMMATION. 2016 9 1038 27 CLINACANTHUS NUTANS EXTRACTS MODULATE EPIGENETIC LINK TO CYTOSOLIC PHOSPHOLIPASE A2 EXPRESSION IN SH-SY5Y CELLS AND PRIMARY CORTICAL NEURONS. CLINACANTHUS NUTANS LINDAU (C. NUTANS), COMMONLY KNOWN AS SABAH SNAKE GRASS IN SOUTHEAST ASIA, IS WIDELY USED IN FOLK MEDICINE DUE TO ITS ANALGESIC, ANTIVIRAL, AND ANTI-INFLAMMATORY PROPERTIES. OUR RECENT STUDY PROVIDED EVIDENCE FOR THE REGULATION OF CYTOSOLIC PHOSPHOLIPASE A2 (CPLA2) MRNA EXPRESSION BY EPIGENETIC FACTORS (TAN ET AL. IN MOL NEUROBIOL. DOI: 10.1007/S12035-015-9314-Z , 2015). THIS ENZYME CATALYZES THE RELEASE OF ARACHIDONIC ACID FROM GLYCEROPHOSPHOLIPIDS, AND FORMATION OF PRO-INFLAMMATORY EICOSANOIDS OR TOXIC LIPID PEROXIDATION PRODUCTS SUCH AS 4-HYDROXYNONENAL. IN THIS STUDY, WE EXAMINED THE EFFECTS OF C. NUTANS ETHANOL LEAF EXTRACTS ON EPIGENETIC REGULATION OF CPLA2 MRNA EXPRESSION IN SH-SY5Y HUMAN NEUROBLASTOMA CELLS AND MOUSE PRIMARY CORTICAL NEURONS. C. NUTANS MODULATED INDUCTION OF CPLA2 EXPRESSION IN SH-SY5Y CELLS BY HISTONE DEACETYLASE (HDAC) INHIBITORS, MS-275, MC-1568, AND TSA. C. NUTANS EXTRACTS ALSO INHIBITED HISTONE ACETYLASE (HAT) ACTIVITY. LEVELS OF CPLA2 MRNA EXPRESSION WERE INCREASED IN PRIMARY CORTICAL NEURONS SUBJECTED TO 0.5-H OXYGEN-GLUCOSE DEPRIVATION INJURY (OGD). THIS INCREASE WAS SIGNIFICANTLY INHIBITED BY C. NUTANS TREATMENT. TREATMENT OF PRIMARY NEURONS WITH THE HDAC INHIBITOR MS-275 AUGMENTED OGD-INDUCED CPLA2 MRNA EXPRESSION, AND THIS INCREASE WAS MODULATED BY C. NUTANS EXTRACTS. OGD-STIMULATED INCREASE IN CPLA2 MRNA EXPRESSION WAS ALSO REDUCED BY A TIP60 HAT INHIBITOR, NU9056. IN VIEW OF A KEY ROLE OF CPLA2 IN THE PRODUCTION OF PRO-INFLAMMATORY EICOSANOIDS AND FREE RADICAL DAMAGE, AND THE FACT THAT EPIGENETIC EFFECTS ON GENES ARE OFTEN LONG-LASTING, RESULTS SUGGEST A ROLE FOR C. NUTANS AND PHYTOCHEMICALS TO INHIBIT THE PRODUCTION OF ARACHIDONIC ACID-DERIVED PRO-INFLAMMATORY EICOSANOIDS AND CHRONIC INFLAMMATION, THROUGH EPIGENETIC REGULATION OF CPLA2 EXPRESSION. 2016 10 5298 23 PROTEIN ARGININE METHYLTRANSFERASE 5 SUPPRESSES THE TRANSCRIPTION OF THE RB FAMILY OF TUMOR SUPPRESSORS IN LEUKEMIA AND LYMPHOMA CELLS. THE PROPER EPIGENETIC MODIFICATION OF CHROMATIN BY PROTEIN ARGININE METHYLTRANSFERASES (PRMTS) IS CRUCIAL FOR NORMAL CELL GROWTH AND HEALTH. THE HUMAN SWI/SNF-ASSOCIATED PRMT5 IS INVOLVED IN THE TRANSCRIPTIONAL REPRESSION OF TARGET GENES BY DIRECTLY METHYLATING H3R8 AND H4R3. TO FURTHER UNDERSTAND THE IMPACT OF PRMT5-MEDIATED HISTONE METHYLATION ON CANCER, WE ANALYZED ITS EXPRESSION IN NORMAL AND TRANSFORMED HUMAN B LYMPHOCYTES. OUR FINDINGS REVEAL THAT PRMT5 PROTEIN LEVELS ARE ENHANCED IN VARIOUS HUMAN LYMPHOID CANCER CELLS, INCLUDING TRANSFORMED CHRONIC LYMPHOCYTIC LEUKEMIA (B-CLL) CELL LINES. PRMT5 OVEREXPRESSION IS CAUSED BY THE ALTERED EXPRESSION OF THE PRMT5-SPECIFIC MICRORNAS 19A, 25, 32, 92, 92B, AND 96 AND RESULTS IN THE INCREASED GLOBAL SYMMETRIC METHYLATION OF H3R8 AND H4R3. AN EVALUATION OF BOTH EPIGENETIC MARKS AT PRMT5 TARGET GENES SUCH AS RB1 (P105), RBL1 (P107), AND RBL2 (P130) SHOWED THAT PROMOTERS H3R8 AND H4R3 ARE HYPERMETHYLATED, WHICH IN TURN TRIGGERS POCKET PROTEIN TRANSCRIPTIONAL REPRESSION. FURTHERMORE, REDUCING PRMT5 EXPRESSION IN WAC3CD5 B-CLL CELLS ABOLISHES H3R8 AND H4R3 HYPERMETHYLATION, RESTORES RBL2 EXPRESSION, AND INHIBITS CANCER CELL PROLIFERATION. THESE RESULTS INDICATE THAT PRMT5 OVEREXPRESSION EPIGENETICALLY ALTERS THE TRANSCRIPTION OF KEY TUMOR SUPPRESSOR GENES AND SUGGEST A CAUSAL ROLE OF THE ELEVATED SYMMETRIC METHYLATION OF H3R8 AND H4R3 AT THE RBL2 PROMOTER IN TRANSFORMED B-LYMPHOCYTE PATHOLOGY. 2008 11 6610 25 UBIQUITIN-SPECIFIC PROTEASE 2-69 IN MACROPHAGES POTENTIALLY MODULATES METAINFLAMMATION. MACROPHAGES PLAY A CRITICAL ROLE IN CHRONIC INFLAMMATION AND METABOLIC DISEASES. WE IDENTIFIED A LONGER SPLICE VARIANT OF UBIQUITIN SPECIFIC PROTEASE (USP) 2-69 AS A NOVEL MOLECULE THAT MODULATES PATHWAYS IMPLICATED IN METABOLIC DISORDERS. EXPRESSION LEVELS OF AP2/FABP4 AND PAI-1/SERPINE1 GENES WERE INCREASED BY 4- AND 1.8-FOLD, RESPECTIVELY, AFTER SHORT HAIRPIN RNA-MEDIATED KNOCKDOWN (KD) OF THE USP2 GENE, AND SUCH EXPRESSION WAS ALLEVIATED BY OVEREXPRESSION OF USP2-69 IN HUMAN MYELOID CELL LINES. SUPERNATANTS DERIVED FROM USP2-KD CELLS INDUCED IL6 ( APPROXIMATELY 6-FOLD) AND SAA3 ( APPROXIMATELY 15-FOLD) IN 3T3-L1 ADIPOCYTES TO SUGGEST THE ANTI-INFLAMMATORY PROPERTIES OF USP2. IN ADDITION, WE OBSERVED A 30% DECREASE IN THE NUMBER OF MACROPHAGES IN MESENTERIC ADIPOSE TISSUE DERIVED FROM USP2-69 TRANSGENIC MICE FED A HIGH-FAT DIET FOR 14 WK COMPARED WITH THAT IN THEIR C57BL/6 LITTERMATES (P<0.01), WHICH WAS CONSISTENT WITH A APPROXIMATELY 40% DECREASE IN TRANSCRIPTION OF AP2 AND PAI-1. THE AP2 LOCUS EXHIBITED ELEVATED CHROMATIN ACCESSIBILITY (>2.1-FOLD), METHYLATION OF HISTONE H3 LYSINE 4 (>4.5-FOLD), AND ACETYLATION OF HISTONE H4 (>2.5-FOLD) IN USP2-KD CELLS. TRANSFECTION OF ISOPEPTIDASE-MUTATED USP2-69 DID NOT ALTER CHROMATIN CONFORMATION ON THE AP2 LOCUS IN USP2-KD CELLS. OUR RESULTS SUGGEST THAT USP2-69 SUPPRESSES META-INFLAMMATORY MOLECULES INVOLVED IN THE DEVELOPMENT OF TYPE-2 DIABETES. 2013 12 2134 29 EPIGENETIC INACTIVATION OF THE MIR129-2 IN HEMATOLOGICAL MALIGNANCIES. BACKGROUND: MIR129-2 HAS BEEN SHOWN TO BE A TUMOR SUPPRESSOR MICRORNA HYPERMETHYLATED IN EPITHELIAL CANCERS. PATIENTS AND METHODS: EPIGENETIC INACTIVATION OF MIR129-2 WAS STUDIED BY METHYLATION-SPECIFIC PCR (MSP) IN 13 CELL LINES (EIGHT MYELOMA AND FIVE LYMPHOMA), 15 NORMAL CONTROLS AND 344 PRIMARY SAMPLES INCLUDING ACUTE MYELOID LEUKEMIA (AML), ACUTE LYMPHOBLASTIC LEUKEMIA (ALL), CHRONIC MYELOID LEUKEMIA (CML), CHRONIC LYMPHOCYTIC LEUKEMIA (CLL), NON-HODGKIN'S LYMPHOMA (NHL), MULTIPLE MYELOMA (MM) AT DIAGNOSIS, MM AT RELAPSE/PROGRESSION, AND MONOCLONAL GAMMOPATHY OF UNDETERMINED SIGNIFICANCE (MGUS). EXPRESSION OF MIR129 AND ITS TARGET, SOX4, IN CELL LINES WAS MEASURED BEFORE AND AFTER HYPOMETHYLATING TREATMENT AND MIR129 OVEREXPRESSION. MIR129 EXPRESSION WAS CORRELATED WITH MIR129-2 METHYLATION STATUS IN PRIMARY LYMPHOMA SAMPLES. TUMOR SUPPRESSOR FUNCTION OF MIR129 WAS DEMONSTRATED BY MTT AND TRYPAN BLUE EXCLUSION ASSAY AFTER MIR129 OVEREXPRESSION. RESULTS: THE SENSITIVITY OF THE METHYLATED-MSP WAS ONE IN 10(3). DIFFERENT MSP STATUSES, INCLUDING COMPLETE METHYLATION, PARTIAL METHYLATION, AND COMPLETE UNMETHYLATION, WERE VERIFIED BY QUANTITATIVE BISULFITE PYROSEQUENCING. ALL FIVE LYMPHOMA AND SEVEN OF EIGHT MYELOMA CELL LINES SHOWED COMPLETE AND PARTIAL MIR129-2 METHYLATION. IN PRIMARY SAMPLES, MIR129-2 METHYLATION WAS ABSENT IN AML AND CML, BUT DETECTED IN 5% ALL, 45.9% CLL, 49.5% MM AT DIAGNOSIS, AND 59.1% NHL. IN CLL, MIR129-2 METHYLATION ADVERSELY IMPACTED ON SURVIVAL (P=0.004). IN MM, MIR129-2 METHYLATION INCREASED FROM 27.5% MGUS TO 49.5% MM AT DIAGNOSIS AND 41.5% AT RELAPSE/PROGRESSION (P=0.023). IN NHL, MIR129-2 METHYLATION WAS ASSOCIATED WITH MIR124-1 AND MIR203 METHYLATION (P<0.001), AND LOWER MIR129 EXPRESSION (P=0.009). HYPOMETHYLATION TREATMENT OF JEKO-1, HOMOZYGOUSLY METHYLATED FOR MIR129-2, LED TO MIR129-2 DEMETHYLATION AND MIR129 RE-EXPRESSION, WITH DOWNREGULATION OF SOX4 MRNA. MOREOVER, MIR129 OVEREXPRESSION IN BOTH MANTLE CELL LINES, JEKO-1 AND GRANTA-519, INHIBITED CELLULAR PROLIFERATION AND ENHANCED CELL DEATH, WITH CONCOMITANT SOX4 MRNA DOWNREGULATION. CONCLUSIONS: MIR129-2 IS A TUMOR SUPPRESSIVE MICRORNA FREQUENTLY METHYLATED IN LYMPHOID BUT NOT MYELOID MALIGNANCIES, LEADING TO REVERSIBLE MIR129-2 SILENCING. IN CLL, MIR129-2 METHYLATION WAS ASSOCIATED WITH AN INFERIOR SURVIVAL. IN MM, MIR129-2 METHYLATION MIGHT BE ACQUIRED DURING PROGRESSION FROM MGUS TO SYMPTOMATIC MM. IN NHL, MIR129-2 METHYLATION MIGHT COLLABORATE WITH MIR124-1 AND MIR203 METHYLATION IN LYMPHOMAGENESIS. 2013 13 5677 31 SHORT AIP1 (ASK1-INTERACTING PROTEIN-1) ISOFORM LOCALIZES TO THE MITOCHONDRIA AND PROMOTES VASCULAR DYSFUNCTION. OBJECTIVE: VASCULAR ENDOTHELIAL CELLS (ECS) NORMALLY MAINTAIN VASCULAR HOMEOSTASIS AND ARE REGULATED BY PROINFLAMMATORY CYTOKINES AND REACTIVE OXYGEN SPECIES. A HUMAN GENOME-WIDE ASSOCIATION STUDY IDENTIFIED THAT AIP1 (ASK1 [APOPTOSIS SIGNAL-REGULATING KINASE 1]-INTERACTING PROTEIN-1; ALSO IDENTIFIED AS DAB2IP) GENE VARIANTS CONFER SUSCEPTIBILITY TO CARDIOVASCULAR DISEASE, BUT THE UNDERLYING MECHANISM IS UNKNOWN. APPROACH AND RESULTS: WE DETECTED A NORMAL AIP1 FORM (NAMED AIP1A) IN THE HEALTHY AORTA, BUT A SHORTER FORM OF AIP1 (NAMED AIP1B) WAS FOUND IN DISEASED AORTAE THAT CONTAINED ATHEROSCLEROTIC PLAQUES AND GRAFT ARTERIOSCLEROSIS. AIP1B TRANSCRIPTION IN RESTING ECS WAS SUPPRESSED THROUGH EPIGENETIC INHIBITION BY RIF1 (RAP1 [RAS-RELATED PROTEIN 1]-INTERACTING FACTOR 1)/H3K9 (HISTONE H3 LYSINE 9) METHYLTRANSFERASE-MEDIATED H3K9 TRIMETHYLATION, AND THIS INHIBITION WAS RELEASED BY PROINFLAMMATORY CYTOKINES. AIP1A, BUT NOT AIP1B, WAS DOWNREGULATED BY PROTEOLYTIC DEGRADATION THROUGH A SMURF1 (SMAD [SUPPRESSOR OF MOTHERS AGAINST DECAPENTAPLEGIC MISCELLANEOUS] UBIQUITYLATION REGULATORY FACTOR 1)-DEPENDENT PATHWAY IN ECS UNDER INFLAMMATION. THEREFORE, AIP1B WAS THE MAJOR FORM PRESENT DURING INFLAMMATORY CONDITIONS. AIP1B, WHICH LACKS THE N-TERMINAL PLECKSTRIN HOMOLOGY DOMAIN OF AIP1A, LOCALIZED TO THE MITOCHONDRIA AND AUGMENTED TNFALPHA (TUMOR NECROSIS FACTOR ALPHA)-INDUCED MITOCHONDRIAL REACTIVE OXYGEN SPECIES GENERATION AND EC ACTIVATION. AIP1B-ECTG (EC-SPECIFIC AIP1B TRANSGENIC) MICE EXHIBITED AUGMENTED REACTIVE OXYGEN SPECIES PRODUCTION, EC ACTIVATION, AND NEOINTIMA FORMATION IN VASCULAR REMODELING MODELS. CONCLUSIONS: OUR CURRENT STUDY SUGGESTS THAT A SHIFT FROM ANTI-INFLAMMATORY AIP1A TO PROINFLAMMATORY AIP1B DURING CHRONIC INFLAMMATION PLAYS A KEY ROLE IN INFLAMMATORY VASCULAR DISEASES. 2020 14 5487 29 REVERSIBLE ALTERATION IN THE EXPRESSION OF THE GAP JUNCTIONAL PROTEIN CONNEXIN 32 DURING TUMOR PROMOTION IN RAT LIVER AND ITS ROLE DURING CELL PROLIFERATION. ALTHOUGH NUMEROUS BIOCHEMICAL MARKERS CAN IDENTIFY PUTATIVE PRENEOPLASTIC ALTERED HEPATIC FOCI (AHF) IN RAT LIVER, NO CONSISTENT PATTERN OF EXPRESSION DURING HEPATOCARCINOGENESIS HAS EMERGED. USING QUANTITATIVE STEREOLOGIC ANALYSES WE DEMONSTRATED THAT DECREASED EXPRESSION OF THE MAJOR HEPATOCYTE GAP JUNCTION PROTEIN, CONNEXIN 32 (CX32), IN RAT AHF IS A CONSISTENT OBSERVATION IN SEVERAL PROTOCOLS OF MULTISTAGE HEPATOCARCINOGENESIS. THIS CHANGE WAS OBSERVED AFTER INITIATION BY EITHER ETHYLNITROSOUREA (ENU) OR DIETHYLNITROSAMINE (DEN), FOLLOWED BY PROMOTION WITH PHENOBARBITAL (PB), DIOXIN, CHLORENDIC ACID, C.I. SOLVENT YELLOW, OR TAMOXIFEN. AHF GENERATED BY WY-14,643, CIPROFIBRATE, AND A CHOLINE/METHIONINE-DEFICIENT DIETARY REGIMEN ALSO SHOWED DECREASED CX32 EXPRESSION. THE DECREASE OF CX32 IN AHF WAS RAPIDLY REVERSIBLE AFTER WITHDRAWAL OF PB, AND THIS CHANGE PRECEDED A REDUCTION IN PLACENTAL ISOZYME OF GLUTATHIONE-S-TRANSFERASE (GST) EXPRESSION IN THE SAME AHF. WITHIN 20 DAYS OF WITHDRAWAL, FEWER THAN 4% OF GST-POSITIVE AHF WERE CX32 DEFICIENT, WHILE THE VOLUME OF TOTAL AHF DECREASED 30%. CHRONIC PB TREATMENT ALSO RESULTED IN A REVERSIBLE DECREASE IN CX32 SPECIFICALLY IN MID- AND CENTRO-LOBULAR HEPATOCYTES. CONTINUOUS THYMIDINE LABELING DEMONSTRATED THAT CX32 COULD BE UNCOUPLED FROM THE CELL CYCLE, SUGGESTING THAT SOME LIVER PROMOTERS MAY ACT DIRECTLY TO ALTER THE EXPRESSION OF CX32. THESE OBSERVATIONS SUGGEST THAT A DECREASE IN CX32 CONTENT WAS A RELATIVELY COMMON EPIGENETIC CHANGE IN AHF INDUCED DURING HEPATOCARCINOGENESIS BY A NUMBER OF INITIATING AND PROMOTING AGENTS BUT THAT THIS CHANGE WAS NOT SUFFICIENT FOR CARCINOGENESIS. THIS CHANGE, HOWEVER, MAY BE NECESSARY FOR THE MECHANISM(S) OF TUMOR PROMOTION, SINCE CX32-POSITIVE AHF DID NOT PROLIFERATE AS READILY AS CX32-DEFICIENT AHF. 1990 15 6256 23 THE MITOGEN AND STRESS-ACTIVATED PROTEIN KINASE 1 REGULATES THE RAPID EPIGENETIC TAGGING OF DORSAL HORN NEURONS AND NOCIFENSIVE BEHAVIOUR. PHOSPHORYLATION OF HISTONE H3 AT SERINE 10 (P-H3S10) IS A MARKER OF ACTIVE GENE TRANSCRIPTION. USING COGNITIVE MODELS OF NEURAL PLASTICITY, P-H3S10 WAS SHOWN TO BE DOWNSTREAM OF EXTRACELLULAR SIGNAL-REGULATED KINASE (ERK) SIGNALLING IN THE HIPPOCAMPUS. IN THIS STUDY, WE SHOW THAT NOCICEPTIVE SIGNALLING AFTER PERIPHERAL FORMALIN INJECTION INCREASED P-H3S10 EXPRESSION IN THE IPSILATERAL DORSAL HORN. THIS INCREASE WAS MAXIMAL 30 MINUTES AFTER FORMALIN INJECTION AND OCCURRED MAINLY WITHIN P-ERK-POSITIVE NEURONS. SPINAL P-H3S10-ENHANCED EXPRESSION WAS ALSO OBSERVED IN NEUROKININ 1 RECEPTOR (NK1R), C-FOS, AND ZIF268 POSITIVE NEURONS AND WAS INHIBITED BY ABLATION OF SEROTONERGIC DESCENDING CONTROLS. THE MITOGEN AND STRESS-ACTIVATED PROTEIN KINASE 1 (MSK1) IS DOWNSTREAM OF ERK AND CAN INDUCE P-H3S10. WE FOUND THAT, AFTER FORMALIN INJECTION, MOST PHOSPHO-MSK1 (P-MSK1)-POSITIVE CELLS (87% +/- 3%) EXPRESSED P-ERK AND THE MAJORITY OF P-H3S10-POSITIVE CELLS (85% +/- 5%) EXPRESSED P-MSK1. INHIBITION OF ERK ACTIVITY WITH THE MEK INHIBITOR SL327 REDUCED FORMALIN-INDUCED P-ERK, P-MSK1, AND P-H3S10, DEMONSTRATING THAT SPINAL P-MSK1 AND P-H3S10 WERE AT LEAST PARTLY DOWNSTREAM OF ERK SIGNALLING. CRUCIALLY, PHARMACOLOGICAL BLOCKADE OF SPINAL MSK1 ACTIVITY WITH THE NOVEL MSK1 INHIBITOR SB727651A INHIBITED FORMALIN-INDUCED SPINAL P-H3S10 AND NOCIFENSIVE BEHAVIOUR. THESE FINDINGS ARE THE FIRST TO ESTABLISH THE INVOLVEMENT OF P-H3S10 AND ITS MAIN KINASE, MSK1, IN ERK REGULATION OF NOCICEPTION. GIVEN THE GENERAL IMPORTANCE OF ERK SIGNALLING IN PAIN PROCESSING, OUR RESULTS SUGGEST THAT P-H3S10 COULD PLAY A ROLE IN THE RESPONSE TO INJURY. 2016 16 5017 28 PERSISTENT INFECTION OF CULTURED CELLS WITH MOUSE HEPATITIS VIRUS (MHV) RESULTS FROM THE EPIGENETIC EXPRESSION OF THE MHV RECEPTOR. THE A59 STRAIN OF MURINE CORONAVIRUS MOUSE HEPATITIS VIRUS (MHV) CAN CAUSE PERSISTENT INFECTION OF 17C1-1 CELLS AND OTHER MURINE CELL LINES. PERSISTENTLY INFECTED CULTURES RELEASED LARGE AMOUNTS OF VIRUS (10(7) TO 10(8) PFU/ML) AND WERE RESISTANT TO SUPERINFECTION WITH MHV BUT NOT TO INFECTION WITH UNRELATED SEMLIKI FOREST AND VESICULAR STOMATITIS VIRUSES. THE CULTURE MEDIUM FROM PERSISTENTLY INFECTED CULTURES DID NOT CONTAIN A SOLUBLE INHIBITOR SUCH AS INTERFERON THAT PROTECTED UNINFECTED CELLS FROM INFECTION BY MHV OR VESICULAR STOMATITIS VIRUS. THE PERSISTENT INFECTION WAS CURED IF FEWER THAN 100 CELLS WERE TRANSFERRED DURING SUBCULTURING, AND SUCH CURED CULTURES WERE SUSCEPTIBLE TO REINFECTION AND THE REESTABLISHMENT OF PERSISTENT INFECTION. CULTURES OF 17C1-1 CELLS THAT HAD BEEN NEWLY CLONED FROM SINGLE CELLS CONSISTED OF A MIXTURE OF MHV-RESISTANT AND -SUSCEPTIBLE CELLS. 17C1-1/#97 CELLS, WHICH WERE CURED BY SUBCLONING AFTER 97 PASSAGES OF A PERSISTENTLY INFECTED CULTURE OVER A 1-YEAR PERIOD, CONTAINED 5 TO 10% OF THEIR POPULATION AS SUSCEPTIBLE CELLS, WHILE 17C1-1/#402 CELLS, WHICH WERE CURED BY SUBCLONING AFTER 402 PASSAGES OVER A 3-YEAR PERIOD, HAD LESS THAN 1% SUSCEPTIBLE CELLS. SUSCEPTIBILITY TO INFECTION CORRELATED WITH THE EXPRESSION OF MHV RECEPTOR GLYCOPROTEIN (MHVR [BGP1A]). FLUORESCENCE-ACTIVATED CELL SORTER ANALYSIS WITH ANTIBODY TO MHVR SHOWED THAT 17C1-1/#97 CELLS CONTAINED A SMALL FRACTION OF MHVR-EXPRESSING CELLS. THESE MHVR-EXPRESSING CELLS WERE SELECTIVELY ELIMINATED WITHIN 24 H AFTER CHALLENGE WITH MHV-A59, AND PRETREATMENT OF 17C1-1/#97 CELLS WITH MONOCLONAL ANTIBODY CC1, WHICH BINDS TO THE N-TERMINAL DOMAIN OF MHVR, BLOCKED INFECTION. WE CONCLUDE THAT THE SUBPOPULATION OF MHVR-EXPRESSING CELLS WERE INFECTED AND KILLED IN ACUTELY OR PERSISTENTLY INFECTED CULTURES, WHILE THE SUBPOPULATION OF MHVR-NONEXPRESSING CELLS SURVIVED AND PROLIFERATED. THE SUBPOPULATION OF MHVR-NEGATIVE CELLS PRODUCED A SMALL PROPORTION OF PROGENY CELLS THAT EXPRESSED MHVR AND BECAME INFECTED, THEREBY MAINTAINING THE PERSISTENT INFECTION AS A STEADY-STATE CARRIER CULTURE. THUS, IN 17C1-1 CELL CULTURES, THE UNSTABLE OR EPIGENETIC EXPRESSION OF MHVR PERMITTED THE ESTABLISHMENT OF A PERSISTENT, CHRONIC INFECTION. 1995 17 1421 31 DIFFERENTIAL BRAIN ADRA2A AND ADRA2C GENE EXPRESSION AND EPIGENETIC REGULATION IN SCHIZOPHRENIA. EFFECT OF ANTIPSYCHOTIC DRUG TREATMENT. POSTSYNAPTIC ALPHA(2A)-ADRENOCEPTOR DENSITY IS ENHANCED IN THE DORSOLATERAL PREFRONTAL CORTEX (DLPFC) OF ANTIPSYCHOTIC-TREATED SCHIZOPHRENIA SUBJECTS. THIS ALTERATION MIGHT BE DUE TO TRANSCRIPTIONAL ACTIVATION, AND COULD BE REGULATED BY EPIGENETIC MECHANISMS SUCH AS HISTONE POSTTRANSLATIONAL MODIFICATIONS (PTMS). THE AIM OF THIS STUDY WAS TO EVALUATE ADRA2A AND ADRA2C GENE EXPRESSION (CODIFYING FOR ALPHA(2)-ADRENOCEPTOR SUBTYPES), AND PERMISSIVE AND REPRESSIVE HISTONE PTMS AT GENE PROMOTER REGIONS IN THE DLPFC OF SUBJECTS WITH SCHIZOPHRENIA AND MATCHED CONTROLS (N = 24 PAIRS). WE STUDIED THE EFFECT OF ANTIPSYCHOTIC (AP) TREATMENT IN AP-FREE (N = 12) AND AP-TREATED (N = 12) SUBGROUPS OF SCHIZOPHRENIA SUBJECTS AND IN RATS ACUTELY AND CHRONICALLY TREATED WITH TYPICAL AND ATYPICAL ANTIPSYCHOTICS. ADRA2A MRNA EXPRESSION WAS SELECTIVELY UPREGULATED IN AP-TREATED SCHIZOPHRENIA SUBJECTS (+93%) WHEREAS ADRA2C MRNA EXPRESSION WAS UPREGULATED IN ALL SCHIZOPHRENIA SUBJECTS (+53%) REGARDLESS OF ANTIPSYCHOTIC TREATMENT. ACUTE AND CHRONIC CLOZAPINE TREATMENT IN RATS DID NOT ALTER BRAIN CORTEX ADRA2A MRNA EXPRESSION BUT INCREASED ADRA2C MRNA EXPRESSION. BOTH ADRA2A AND ADRA2C PROMOTER REGIONS SHOWED EPIGENETIC MODIFICATION BY HISTONE METHYLATION AND ACETYLATION IN HUMAN DLPFC. THE UPREGULATION OF ADRA2A EXPRESSION IN AP-TREATED SCHIZOPHRENIA SUBJECTS MIGHT BE RELATED TO OBSERVED BIVALENT CHROMATIN AT ADRA2A PROMOTER REGION IN SCHIZOPHRENIA (DEPICTED BY INCREASED PERMISSIVE H3K4ME3 AND REPRESSIVE H3K27ME3) AND COULD BE TRIGGERED BY THE ENHANCED H4K16AC AT ADRA2A PROMOTER. IN CONCLUSION, EPIGENETIC PREDISPOSITION DIFFERENTIALLY MODULATED ADRA2A AND ADRA2C MRNA EXPRESSION IN DLPFC OF SCHIZOPHRENIA SUBJECTS. 2021 18 4527 24 MULTIGENERATIONAL EFFECTS OF 4-METHYLBENZYLIDENE CAMPHOR (4-MBC) ON THE SURVIVAL, DEVELOPMENT AND REPRODUCTION OF THE MARINE COPEPOD TIGRIOPUS JAPONICUS. ONE OF THE MOST WIDELY USED ORGANIC UV FILTERS, 4-METHYLBENZYLIDENE CAMPHOR (4-MBC), IS PRESENT AT HIGH CONCENTRATIONS IN OFFSHORE WATERS. THE MARINE COPEPOD TIGRIOPUS JAPONICUS WAS EXPOSED TO DIFFERENT CONCENTRATIONS OF 4-MBC (I.E., 0, 0.5, 1, 5 AND 10MUGL(-1)) FOR 4 CONSECUTIVE GENERATIONS (F0-F3) TO EVALUATE THE IMPACT OF 4-MBC ON MARINE ECOSYSTEMS. THE RESULTS SHOWED THAT IN THE F0 GENERATION, 4-MBC CAUSED SIGNIFICANT LETHAL TOXICITY IN T. JAPONICAS AT CONCENTRATIONS OF 5 AND 10MUGL(-1) AND THE NAUPLII WERE MORE SENSITIVE TO 4-MBC TOXICITY THAN THE ADULTS. HOWEVER IN THE F1-F3 GENERATIONS, 4-MBC EXPOSURE DID NOT AFFECT THE SURVIVAL RATE. THE HATCHING RATE AND THE DEVELOPMENTAL DURATION FROM THE NAUPLII TO THE COPEPODITE (N-C) AND FROM THE NAUPLII TO ADULT (N-A) DECREASED SIGNIFICANTLY IN THE F1-F2 GENERATIONS AND IN THE F2-F3 GENERATIONS, RESPECTIVELY, EVEN AT THE LOWEST EXPOSURE CONCENTRATION (0.5MUGL(-1)). IN THE SUBSEQUENT TWO GENERATIONS (I.E., THE F4-F5 GENERATIONS) OF RECOVERY EXPOSURE IN CLEAN SEAWATER, THE GROWTH RATES OF THE ORIGINAL 4-MBC EXPOSURE GROUPS WERE STILL FASTER THAN THE CONTROL IN BOTH THE N-C AND N-A STAGES, SUGGESTING POSSIBLE TRANSGENERATIONAL GENETIC AND/OR EPIGENETIC CHANGES UPON CHRONIC 4-MBC EXPOSURE. THE EXPRESSION OF THE ECDYSONE RECEPTOR GENE WAS UP-REGULATED BY 4-MBC, WHICH WAS CONSISTENT WITH THE DECREASE OF THE N-C/N-A DURATION. IN ADDITION, 4-MBC MAY INDUCE OXIDATIVE STRESS AND TRIGGER APOPTOSIS IN T. JAPONICAS, RESULTING IN DEVELOPMENTAL, REPRODUCTIVE AND EVEN LETHAL TOXICITY. A PRELIMINARY RISK ASSESSMENT SUGGESTED THAT UNDER ENVIRONMENTALLY REALISTIC CONCENTRATIONS, 4-MBC HAD SIGNIFICANT POTENTIAL TO POSE A THREAT TO MARINE CRUSTACEANS AND MARINE ECOSYSTEMS. 2018 19 3303 21 HIGH-FREQUENCY P16(INK) (4A) PROMOTER METHYLATION IS ASSOCIATED WITH HISTONE METHYLTRANSFERASE SETDB1 EXPRESSION IN SPORADIC CUTANEOUS MELANOMA. EPIGENETIC MECHANISMS PARTICIPATE IN MELANOMA DEVELOPMENT AND PROGRESSION. THE EFFECT OF HISTONE MODIFICATIONS AND THEIR CATALYSING ENZYMES OVER EUCHROMATIC PROMOTER DNA METHYLATION IN MELANOMA REMAINS UNCLEAR. THIS STUDY INVESTIGATED THE POTENTIAL ASSOCIATION OF P16(INK) (4A) PROMOTER METHYLATION WITH HISTONE METHYLTRANSFERASE SETDB1 EXPRESSION IN GREEK PATIENTS WITH SPORADIC MELANOMA AND THEIR CORRELATION WITH CLINICOPATHOLOGICAL CHARACTERISTICS. PROMOTER METHYLATION WAS DETECTED BY METHYLATION-SPECIFIC PCR IN 100 PERIPHERAL BLOOD SAMPLES AND 58 MELANOMA TISSUES FROM THE SAME PATIENTS. CELL PROLIFERATION (KI-67 INDEX), P16(INK) (4A) AND SETDB1 EXPRESSION WERE EVALUATED BY IMMUNOHISTOCHEMISTRY. HIGH-FREQUENCY PROMOTER METHYLATION (25.86%) WAS OBSERVED IN TISSUE SAMPLES AND CORRELATED WITH INCREASED CELL PROLIFERATION (P = 0.0514). P16(INK) (4A) PROMOTER METHYLATION WAS HIGHER IN VERTICAL GROWTH-PHASE (60%) MELANOMAS THAN IN RADIAL (40%, P = 0.063) AND THOSE DISPLAYING EPIDERMAL INVOLVEMENT (P = 0.046). IMPORTANTLY, P16(INK) (4A) METHYLATION CORRELATED WITH INCREASED MELANOMA THICKNESS ACCORDING TO BRESLOW INDEX (P = 0.0495) AND MARGINALLY WITH INCREASED CLARK LEVEL (I/II VS III/IV/V, P = 0.070). LOW (1-30%) P16(INK) (4A) EXPRESSION WAS DETECTED AT THE MAJORITY (19 OF 54) OF MELANOMA CASES (35.19%), BEING MARGINALLY CORRELATED WITH TUMOR LYMPHOCYTIC INFILTRATION (P = 0.078). SETDB1 NUCLEAR IMMUNOREACTIVITY WAS OBSERVED IN 47 OF 57 (82.46%) CASES, WHEREAS 27 OF 57 (47.37%) SHOWED CYTOPLASMIC IMMUNOEXPRESSION. CYTOPLASMIC SETDB1 EXPRESSION CORRELATED WITH HIGHER FREQUENCY OF P16(INK) (4A) METHYLATION AND P16(INK) (4A) EXPRESSION (P = 0.033, P = 0.011, RESPECTIVELY). INCREASED NUCLEAR SETDB1 LEVELS WERE ASSOCIATED WITH HIGHER MITOTIC COUNT (0-5/MM(2) VS >5/MM(2) , P = 0.0869), ADVANCED CLARK LEVEL (III-V, P = 0.0380), EPIDERMAL INVOLVEMENT (P = 0.0331) AND THE NON-CHRONIC SUN EXPOSURE-ASSOCIATED MELANOMA TYPE (P = 0.0664). OUR DATA DEMONSTRATE FOR THE FIRST TIME THE ASSOCIATION OF HISTONE METHYLTRANSFERASE SETDB1 WITH FREQUENT METHYLATION OF THE EUCHROMATIC P16(INK) (4A) PROMOTER AND SEVERAL PROGNOSTIC PARAMETERS IN MELANOMAS. 2014 20 2080 26 EPIGENETIC DNA METHYLATION OF EBI3 MODULATES HUMAN INTERLEUKIN-35 FORMATION VIA NFKB SIGNALING: A PROMISING THERAPEUTIC OPTION IN ULCERATIVE COLITIS. ULCERATIVE COLITIS (UC), A SEVERE CHRONIC DISEASE WITH UNCLEAR ETIOLOGY THAT IS ASSOCIATED WITH INCREASED RISK FOR COLORECTAL CANCER, IS ACCOMPANIED BY DYSREGULATION OF CYTOKINES. EPSTEIN-BARR VIRUS-INDUCED GENE 3 (EBI3) ENCODES A SUBUNIT IN THE UNIQUE HETERODIMERIC IL-12 CYTOKINE FAMILY OF EITHER PRO- OR ANTI-INFLAMMATORY FUNCTION. AFTER HAVING RECENTLY DEMONSTRATED THAT UPREGULATION OF EBI3 BY HISTONE ACETYLATION ALLEVIATES DISEASE SYMPTOMS IN A DEXTRAN SULFATE SODIUM (DSS)-TREATED MOUSE MODEL OF CHRONIC COLITIS, WE NOW AIMED TO EXAMINE A POSSIBLE FURTHER EPIGENETIC REGULATION OF EBI3 BY DNA METHYLATION UNDER INFLAMMATORY CONDITIONS. TREATMENT WITH THE DNA METHYLTRANSFERASE INHIBITOR (DNMTI) DECITABINE (DAC) AND TNFALPHA LED TO SYNERGISTIC UPREGULATION OF EBI3 IN HUMAN COLON EPITHELIAL CELLS (HCEC). USE OF DIFFERENT SIGNALING PATHWAY INHIBITORS INDICATED NFKAPPAB SIGNALING WAS NECESSARY AND PROPORTIONAL TO THE SYNERGISTIC EBI3 INDUCTION. MALDI-TOF/MS AND HPLC-ESI-MS/MS ANALYSIS OF DAC/TNFALPHA-TREATED HCEC IDENTIFIED IL-12P35 AS THE MOST PROBABLE BINDING PARTNER TO FORM A FUNCTIONAL PROTEIN. EBI3/IL-12P35 HETERODIMERS (IL-35) INDUCE THEIR OWN GENE UPREGULATION, SOMETHING THAT WAS INDEED OBSERVED IN HCEC CULTURED WITH MEDIA FROM PREVIOUSLY DAC/TNFALPHA-TREATED HCEC. THESE RESULTS SUGGEST THAT UNDER INFLAMMATORY AND DEMETHYLATING CONDITIONS THE UPREGULATION OF EBI3 RESULTS IN THE FORMATION OF ANTI-INFLAMMATORY IL-35, WHICH MIGHT BE CONSIDERED AS A THERAPEUTIC TARGET IN COLITIS. 2021