1 364 123 AMELIORATION OF UREMIC TOXIN INDOXYL SULFATE-INDUCED OSTEOBLASTIC CALCIFICATION BY SET DOMAIN CONTAINING LYSINE METHYLTRANSFERASE 7/9 PROTEIN. BACKGROUND: VASCULAR CALCIFICATION (VC) IS A VERY COMMON PHENOMENON IN PATIENTS WITH CHRONIC KIDNEY DISEASE (CKD). IT HAS BEEN REPORTED THAT SOME HISTONE METHYLATION PLAY A ROLE IN VC AS AN EPIGENETIC REGULATOR. INDOXYL SULFATE (IS) IS A PROTEIN-BOUND UREMIC TOXIN THAT HAS BEEN PROVEN AS ONE OF THE MAJOR RISK FACTORS OF CARDIOVASCULAR DISEASE IN CKD. SET DOMAIN CONTAINING LYSINE METHYLTRANSFERASE 7/9 (SET7/9) IS ONE OF THE IMPORTANT HISTONE METHYLTRANSFERASES. OBJECTIVES: THIS STUDY AIMED TO DETERMINE THE EFFECT OF IS ON THE EXPRESSION OF SET7/9 AND THE ROLE OF SET7/9 IN IS-INDUCED OSTEOBLASTIC DIFFERENTIATION AND CALCIFICATION OF VASCULAR SMOOTH MUSCLE CELLS (VSMCS). METHODS: VSMCS WERE INCUBATED WITH VARIOUS CONCENTRATIONS OF IS FOR DIFFERENT DURATIONS TO ASSESS OSTEOBLASTIC DIFFERENTIATION AND EXPRESSION OF SET7/9. WESTERN BLOT ANALYSIS AND QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION WERE PERFORMED TO ASSESS THE PROTEIN AND MRNA LEVELS OF SET7/9 RESPECTIVELY. THE CALCIUM CONTENT WAS MEASURED TO EVALUATE CALCIFICATION. RESULTS: OSTEOBLASTIC DIFFERENTIATION AND CALCIFICATION OF VSMCS AND DOWNREGULATION OF THE EXPRESSION OF SET7/9 WERE OBSERVED AFTER IS TREATMENT. THE AUTOPHAGY WAS ACTIVATED AFTER TREATMENT WITH IS, WHEREAS THE INHIBITION OF THE AUTOPHAGY PARTIALLY ATTENUATED THE EFFECT OF IS ON BOTH THE STIMULATION OF THE EXPRESSION OF RUNT-RELATED TRANSCRIPTION FACTOR 2 AND CALCIUM DEPOSITION. CONCLUSIONS: OUR DATA DEMONSTRATED THAT SET7/9 DOWNREGULATION AND AUTOPHAGY ACTIVATION MAY BE THE KEY MECHANISM OF IS-INDUCED VC IN CKD. 2019 2 4644 20 NEUROPATHIC PAIN TREATMENT: STILL A CHALLENGE. NEUROPATHIC PAIN (NP) IS THE RESULT OF A SERIES OF CONDITIONS CAUSED BY DISEASES OR LESIONS TO THE SOMATOSENSORY SYSTEM. DUE TO THE BETTER UNDERSTANDING OF NP PATHOPHYSIOLOGY PREVIOUSLY UNEXPLORED THERAPIES HAVE BEEN USED WITH ENCOURAGING RESULTS. IN THIS GROUP, ACETYL-L-CARNITINE, ALPHA-LIPOIC-ACID, CANNABINOIDS, CLONIDINE, EMA401, BOTULINUM TOXIN TYPE A AND NEW VOLTAGE-GATED SODIUM CHANNEL BLOCKERS, CAN BE INCLUDED. BESIDES, CHANGING PARADIGMS MAY OCCUR WITH THE ADVENT OF OPTOGENETICS AND A BETTER UNDERSTANDING OF EPIGENETIC REGULATION. WE REVIEWED THE PUBLISHED LITERATURE ON THE PHARMACOLOGICAL TREATMENT OF NP. DESPITE THE INTERESTING RESULTS, RANDOMIZED CONTROLLED TRIALS ARE DEMANDED THE MAJORITY OF THE THERAPIES PREVIOUSLY MENTIONED. IN SPITE OF SEVERAL STUDIES FOR THE RELIEF OF NP, PAIN CONTROL CONTINUES BEING A CHALLENGE. 2016 3 1881 23 EMERGING TREATMENTS FOR NEUROPATHIC PAIN. NEUROPATHIC PAIN IS A SERIES OF WELL-KNOWN CONDITIONS CAUSED BY DISEASES OR LESIONS TO THE SOMATOSENSORY SYSTEM. DUE TO THE BETTER UNDERSTANDING OF THE PATHOPHYSIOLOGY OF NEUROPATHIC PAIN, PREVIOUSLY UNEXPLORED THERAPIES HAVE BEEN USED WITH ENCOURAGING RESULTS. AS SUCH, ACETYL-L-CARNITINE (ALC), ALPHA-LIPOIC-ACID (ALA), CANNABINOIDS, CLONIDINE, EMA401, BOTULINUM TOXIN TYPE A, AND NEW VOLTAGE-GATED SODIUM CHANNEL BLOCKERS, CAN BE CITED. FURTHERMORE, NEW MODALITIES IN NEUROMODULATION SUCH AS HIGH-FREQUENCY SPINAL CORD STIMULATION, BURST STIMULATION, DORSAL ROOT GANGLION STIMULATION, TRANSCRANIAL DIRECT CURRENT STIMULATION, AND MANY OTHERS HAVE BEEN SHOWING EXCITING RESULTS. BESIDES, CHANGING PARADIGMS MAY OCCUR WITH THE ADVENT OF OPTOGENETICS AND A BETTER UNDERSTANDING OF EPIGENETIC REGULATION. THIS ARTICLE REVIEWS THE PUBLISHED LITERATURE ON THE TREATMENT OF NP. DESPITE THE INTERESTING RESULTS, RANDOMIZED CONTROLLED TRIALS ARE DEMANDED FOR THE MAJORITY OF THE THERAPIES PREVIOUSLY MENTIONED. 2015 4 3654 64 INDOXYL SULFATE ACCELERATES VASCULAR SMOOTH MUSCLE CELL CALCIFICATION VIA MICRORNA-29B DEPENDENT REGULATION OF WNT/BETA-CATENIN SIGNALING. VASCULAR CALCIFICATION (VC) IS A VERY COMMON PHENOMENON IN PATIENTS WITH CHRONIC KIDNEY DISEASE(CKD) AND IT INCREASES THE INCIDENCE OF CARDIOVASCULAR DISEASE AND LEADS TO HIGH MORTALITY IN CKD PATIENTS. IT HAS BEEN REPORTED THAT SOME MICRORNAS (MIRS) PLAY ROLES IN VASCULAR CALCIFICATION AS AN EPIGENETIC REGULATOR. INDOXYL SULFATE (IS) IS A PROTEIN-BOUND UREMIC TOXIN WHICH HAS BEEN PROVEN AS ONE OF THE MAJOR RISK FACTORS OF CARDIOVASCULAR DISEASE IN CKD. HERE WE INVESTIGATED WHETHER MICRORNA-29B (MIR-29B) IS INVOLVED IN IS-INDUCED VASCULAR CALCIFICATION. WE FOUND THAT VASCULAR MIR-29B WAS DOWN-REGULATED IN RADIAL ARTERIES OF PATIENTS WITH END-STAGE RENAL DISEASE. CONSISTENTLY, IS ALSO DECREASED MIR-29B EXPRESSION IN HUMAN AORTIC SMOOTH MUSCLE CELLS (HASMCS) AND POTENTIATED THEIR CALCIFICATION. MIR-29B MIMICS SIGNIFICANTLY SUPPRESSED, WHILE MIR-29B ANTI-MIR MARKEDLY ENHANCED, IS-INDUCED RUNT-RELATED TRANSCRIPTION FACTOR 2 AND OSTEOPONTIN EXPRESSION. THE EXPRESSION OF WNT7B/BETA-CATENIN IN RADIAL ARTERIES WAS HIGHER IN END STAGE RENAL DISEASE THAN IN CONTROL GROUP, AND IS INCREASED WNT7B/BETA-CATENIN EXPRESSION IN HASMCS AS EARLY AS 3DAYS AFTER STIMULATION. FURTHERMORE, MIR-29B MIMICS POTENTLY REPRESSED WNT7B/BETA-CATENIN PROTEIN EXPRESSION IN HASMCS, WHEREAS MIR-29B ANTI-MIR INCREASED THEIR EXPRESSION, INDICATING MIR-29B INDEED NEGATIVELY REGULATES WNT7B/BETA-CATENIN SIGNALING. DICKKOPF-1 PROTEIN, THE WNT/BETA-CATENIN SIGNALING INHIBITOR, SUPPRESSED ANTI-MIR-29B-ENHANCED HASMCS CALCIFICATION. OUR DATA THUS INDICATE THAT MIR-29B DOWNREGULATION AND WNT/BETA-CATENIN SIGNALING ACTIVATION MAY BE THE KEY MECHANISM OF IS INDUCED VASCULAR CALCIFICATION IN CHRONIC KIDNEY DISEASE. 2018 5 2358 28 EPIGENETIC REGULATION OF RAC1 INDUCES SYNAPTIC REMODELING IN STRESS DISORDERS AND DEPRESSION. DEPRESSION INDUCES STRUCTURAL AND FUNCTIONAL SYNAPTIC PLASTICITY IN BRAIN REWARD CIRCUITS, ALTHOUGH THE MECHANISMS PROMOTING THESE CHANGES AND THEIR RELEVANCE TO BEHAVIORAL OUTCOMES ARE UNKNOWN. TRANSCRIPTIONAL PROFILING OF THE NUCLEUS ACCUMBENS (NAC) FOR RHO GTPASE-RELATED GENES, WHICH ARE KNOWN REGULATORS OF SYNAPTIC STRUCTURE, REVEALED A SUSTAINED REDUCTION IN RAS-RELATED C3 BOTULINUM TOXIN SUBSTRATE 1 (RAC1) EXPRESSION AFTER CHRONIC SOCIAL DEFEAT STRESS. THIS WAS ASSOCIATED WITH A REPRESSIVE CHROMATIN STATE SURROUNDING THE PROXIMAL PROMOTER OF RAC1. INHIBITION OF CLASS 1 HISTONE DEACETYLASES (HDACS) WITH MS-275 RESCUED BOTH THE DECREASE IN RAC1 TRANSCRIPTION AFTER SOCIAL DEFEAT STRESS AND DEPRESSION-RELATED BEHAVIOR, SUCH AS SOCIAL AVOIDANCE. WE FOUND A SIMILAR REPRESSIVE CHROMATIN STATE SURROUNDING THE RAC1 PROMOTER IN THE NAC OF SUBJECTS WITH DEPRESSION, WHICH CORRESPONDED WITH REDUCED RAC1 TRANSCRIPTION. VIRAL-MEDIATED REDUCTION OF RAC1 EXPRESSION OR INHIBITION OF RAC1 ACTIVITY IN THE NAC INCREASES SOCIAL DEFEAT-INDUCED SOCIAL AVOIDANCE AND ANHEDONIA IN MICE. CHRONIC SOCIAL DEFEAT STRESS INDUCES THE FORMATION OF STUBBY EXCITATORY SPINES THROUGH A RAC1-DEPENDENT MECHANISM INVOLVING THE REDISTRIBUTION OF SYNAPTIC COFILIN, AN ACTIN-SEVERING PROTEIN DOWNSTREAM OF RAC1. OVEREXPRESSION OF CONSTITUTIVELY ACTIVE RAC1 IN THE NAC OF MICE AFTER CHRONIC SOCIAL DEFEAT STRESS REVERSES DEPRESSION-RELATED BEHAVIORS AND PRUNES STUBBY SPINES. TAKEN TOGETHER, OUR DATA IDENTIFY EPIGENETIC REGULATION OF RAC1 IN THE NAC AS A DISEASE MECHANISM IN DEPRESSION AND REVEAL A FUNCTIONAL ROLE FOR RAC1 IN RODENTS IN REGULATING STRESS-RELATED BEHAVIORS. 2013 6 6612 32 ULTRA-LOW-DOSE NALOXONE ENHANCES THE ANTINOCICEPTIVE EFFECT OF MORPHINE IN PTX-TREATED RATS: REGULATION ON GLOBAL HISTONE METHYLATION. OBJECTIVE: EPIGENETIC REPROGRAMMING MAY HAVE A POSSIBLE ROLE IN NEUROPATHIC PAIN DEVELOPMENT; THE PRESENT STUDY EXAMINED THE GLOBAL PATTERNS OF LYSINE HISTONE MODIFICATION. IN THIS SERIAL STUDY WE ANALYZED THE LEVELS OF HISTONE 3 LYSINE 4 MONOMETHYLATION, HISTONE 3 LYSINE 4 DIMETHYLATION, AND HISTONE 3 LYSINE 9 TRIMETHYLATION IN PERTUSSIS TOXIN (PTX)-INDUCED THERMAL HYPERALGESIC RAT SPINAL CORDS. METHODS: MALE WISTAR RATS IMPLANTED WITH AN INTRATHECAL CATHETER RECEIVED A SINGLE INTRATHECAL PTX (1 MUG IN 5 MUL SALINE) INJECTION. FOUR DAYS LATER, THEY WERE RANDOMLY ASSIGNED TO RECEIVE EITHER A SINGLE INJECTION OF SALINE, OR ULTRA-LOW-DOSE NALOXONE (15 NG IN 5 MUL SALINE), FOLLOWED BY MORPHINE (10 MUG IN 5 MUL SALINE) INJECTION 30 MINUTES LATER. RESULTS: THE RESULTS SHOWED THAT PTX INJECTION INDUCED THERMAL HYPERALGESIA AND SIGNIFICANT INCREASE OF GLOBAL HISTONE METHYLATION IN THE SPINAL CORDS. INTRATHECAL MORPHINE ALONE DID NOT AFFECT THE THERMAL HYPERALGESIA AND GLOBAL HISTONE METHYLATION. IN CONTRAST, INTRATHECAL ADMINISTRATION OF ULTRA-LOW-DOSE NALOXONE PLUS MORPHINE SIGNIFICANTLY ATTENUATED THE PTX-INDUCED THERMAL HYPERALGESIA AND DOWN-REGULATED THE GLOBAL HISTONE METHYLATION. CONCLUSION: THE RESULTS SUGGEST THAT ULTRA-LOW-DOSE NALOXONE MIGHT BE CLINICAL VALUABLE FOR NEUROPATHIC PAIN MANAGEMENT VIA REGULATING GLOBAL HISTONE MODIFICATION. 2012 7 3655 37 INDOXYL SULFATE ENHANCE THE HYPERMETHYLATION OF KLOTHO AND PROMOTE THE PROCESS OF VASCULAR CALCIFICATION IN CHRONIC KIDNEY DISEASE. CHRONIC KIDNEY DISEASE (CKD) IS A STATE OF KLOTHO DEFICIENCY. THE KLOTHO EXPRESSION MAY BE SUPPRESSED DUE TO DNA HYPERMETHYLATION IN CANCER CELLS SO WE HAVE INVESTIGATED THE EFFECTS AND POSSIBLE MECHANISMS BY WHICH KLOTHO EXPRESSION IS REGULATED IN HUMAN AORTIC SMOOTH MUSCLE CELLS (HASMCS). THE VASCULAR KLOTHO HYPERMETHYLATION IN RADIAL ARTERIES OF PATIENTS WITH END-STAGE RENAL DISEASE WAS DESCRIBED. CULTURED HASMCS AND 5/6-NEPHRECTOMIZED SPRAGUE DAWLEY (SD) RATS TREATED WITH INDOXYL SULFATE (IS) WERE USED AS IN VITRO AND IN VIVO MODELS, RESPECTIVELY. IS INCREASED CPG HYPERMETHYLATION OF THE KLOTHO GENE AND DECREASED KLOTHO EXPRESSION IN HASMCS, AND POTENTIATED HASMCS CALCIFICATION. THE EXPRESSION OF DNA METHYLTRANSFERASE (DNMT) 1 AND 3A IN HASMCS TREATED WITH IS WAS SIGNIFICANTLY INCREASED AND SPECIFIC INHIBITION OF DNA METHYLTRANSFERASE 1 BY 5-AZA-2'-DEOXYCYTIDINE(5AZA-2DC) CAUSED DEMETHYLATION OF THE KLOTHO GENE AND INCREASED KLOTHO EXPRESSION. IN RATS, INJECTION OF IS POTENTIATED VASCULAR CALCIFICATION, INCREASED CPG HYPERMETHYLATION OF THE KLOTHO GENE AND DECREASED KLOTHO EXPRESSION IN THE AORTIC MEDIAL LAYER AND ALL OF THESE CHANGES COULD BE REVERTED BY 5AZA-2DC TREATMENT. TRANSCRIPTIONAL SUPPRESSION OF VASCULAR KLOTHO GENE EXPRESSION BY IS AND EPIGENETIC MODIFICATION OF KLOTHO BY IS MAY BE AN IMPORTANT PATHOLOGICAL MECHANISM OF VASCULAR CALCIFICATION IN CKD. 2016 8 1850 28 ELECTROACUPUNCTURE AMELIORATES DEPRESSION-LIKE BEHAVIORS COMORBID TO CHRONIC NEUROPATHIC PAIN VIA TET1-MEDIATED RESTORATION OF ADULT NEUROGENESIS. ALTHOUGH ELECTROACUPUNCTURE (EA) STIMULATION IS A WIDELY USED THERAPY FOR CHRONIC PAIN AND COMORBID PSYCHIATRIC DISORDERS, ITS LONG-TERM EFFECTS ON CHRONIC NEUROPATHIC PAIN-INDUCED DEPRESSION AND THE UNDERLYING MECHANISMS REMAIN ELUSIVE. IN THE PRESENT STUDY, WE FOUND THAT EA STIMULATION WAS ABLE TO RESTORE ADULT NEUROGENESIS IN THE VENTRAL DENTATE GYRUS (DG), BY BOTH INCREASING NEURONAL DIFFERENTIATION AND RESTORING THE NORMAL MORPHOLOGY OF NEWBORN DENDRITES, IN MICE WITH SPARED NERVE INJURY SURGERY. BY ABLATING THE NESTIN+ NEURAL STEM CELLS (NSCS) VIA DIPHTHERIA TOXIN FRAGMENT A EXPRESSION, WE FURTHER PROVED THAT NEUROGENESIS IN THE VENTRAL DG WAS CRUCIAL TO THE LONG-TERM, BUT NOT THE IMMEDIATE ANTIDEPRESSANT EFFECT OF EA, NOR WAS IT ASSOCIATED WITH NOCICEPTION. FURTHERMORE, WE FOUND THAT THE RESTORATION OF NEUROGENESIS WAS DEPENDENT ON TET1-MEDIATED EPIGENETIC MODIFICATION UPON EA TREATMENT. TET1 COULD BIND TO THE PROMOTER OF THE PROX1 GENE, THUS CATALYZING ITS DEMETHYLATION AND FACILITATING ITS EXPRESSION, WHICH FINALLY CONTRIBUTED TO THE RESTORATION OF NEUROGENESIS AND AMELIORATION OF DEPRESSION-LIKE BEHAVIORS INDUCED BY CHRONIC NEUROPATHIC PAIN. THUS, WE CONCLUDE THAT EA STIMULATION RESTORES INHIBITED TET1 EXPRESSION IN HIPPOCAMPAL NSCS OF MICE WITH CHRONIC NEUROPATHIC PAIN, AND INCREASED TET1 EXPRESSION AMELIORATES HYPERMETHYLATION OF PROX1 AND RESTORES NORMAL ADULT NEUROGENESIS IN THE VENTRAL DG, WHICH CONTRIBUTES TO THE LONG-TERM ANTIDEPRESSANT EFFECT OF EA. 2023 9 3887 28 KLOTHO METHYLATION IS LINKED TO UREMIC TOXINS AND CHRONIC KIDNEY DISEASE. EPIGENETIC REGULATION PLAYS A MAJOR ROLE IN UREMIC TOXIN-INDUCED CHRONIC KIDNEY DISEASE (CKD) PROGRESSION. THE KLOTHO PROTEIN IS A KEY MODULATOR OF HOMEOSTASIS IN RENAL FUNCTION. UREMIC TOXIN ACCUMULATION CAN INDUCE DNA METHYLTRANSFERASE (DNMT) PROTEIN EXPRESSION, WHICH IS INVOLVED IN THE SILENCING OF KLOTHO THROUGH HYPERMETHYLATION. TREATMENT WITH DNMT INHIBITORS CAN INDUCE A HYPERMETHYLATED STATUS OF KLOTHO AND SUPPRESS MRNA AND PROTEIN EXPRESSION. EPIGENETIC TARGETING OF SPECIFIC GENES MAY BECOME AN EFFECTIVE STRATEGY TO PREVENT PROGRESSION OF UREMIA-RELATED CKD. 2012 10 1980 27 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 11 1800 32 EFFECT OF HISTONE DEACETYLASE INHIBITOR ON ETHANOL WITHDRAWAL-INDUCED HYPERALGESIA IN RATS. BACKGROUND: INCREASED PAIN SENSITIVITY IS OBSERVED FOLLOWING ALCOHOL WITHDRAWAL, AND ATTEMPTS TO ALLEVIATE THIS HYPERALGESIA CAN CONTRIBUTE TO THE CYCLE OF ADDICTION. THE AIM OF THIS STUDY WAS TO DETERMINE IF ALCOHOL WITHDRAWAL-INDUCED HYPERALGESIA WAS OBSERVED IN A CHRONIC ETHANOL EXPOSURE MODEL AND IF THIS PAIN WAS AFFECTED BY HISTONE DEACETYLASE INHIBITORS, THUS REVEALING AN EPIGENETIC MECHANISM. METHODS: ADULT MALE SPRAGUE DAWLEY RATS RECEIVED LIEBER-DECARLI LIQUID CONTROL OR ETHANOL (9% V/V) DIET FOR 15 DAYS. MECHANICAL SENSITIVITY WAS MEASURED WITH VON FREY HAIR STIMULATION OF THE HINDPAW DURING ETHANOL ADMINISTRATION AND 24- AND 72-HOUR WITHDRAWAL. RESULTS: ETHANOL WITHDRAWAL PRODUCED SEVERE AND SUSTAINED MECHANICAL HYPERALGESIA, AN EFFECT NOT OBSERVED IN THE CONTROL OR ETHANOL-MAINTAINED GROUPS. FURTHERMORE, THIS HYPERALGESIA WAS ATTENUATED BY THE HISTONE DEACETYLASE INHIBITOR, SUBEROYLANILIDE HYDROXAMIC ACID TREATMENT. CONCLUSIONS: HEIGHTENED PAIN SENSITIVITY WAS OBSERVED FOLLOWING WITHDRAWAL FROM CHRONIC ETHANOL EXPOSURE, AND HISTONE DEACETYLASE INHIBITORS COULD BE NOVEL TREATMENTS FOR THIS ALCOHOL WITHDRAWAL-INDUCED HYPERALGESIA. 2019 12 872 33 CHRONIC ALCOHOL EXPOSURE DIFFERENTIALLY ALTERS ONE-CARBON METABOLISM IN RAT LIVER AND BRAIN. BACKGROUND: EPIGENETIC MECHANISMS SUCH AS DNA METHYLATION PLAY AN IMPORTANT ROLE IN REGULATING THE PATHOPHYSIOLOGY OF ALCOHOLISM. CHRONIC ALCOHOL EXPOSURE LEADS TO BEHAVIORAL CHANGES AS WELL AS DECREASED EXPRESSION OF GENES ASSOCIATED WITH SYNAPTIC PLASTICITY. IN THE LIVER, IT HAS BEEN DOCUMENTED THAT CHRONIC ALCOHOL EXPOSURE IMPAIRS METHIONINE SYNTHASE (MS) ACTIVITY LEADING TO A DECREASE IN S-ADENOSYL METHIONINE/S-ADENOSYL HOMOCYSTEINE (SAM/SAH) RATIO WHICH RESULTS IN DNA HYPOMETHYLATION; HOWEVER, IT IS NOT KNOWN WHETHER SIMILAR ALTERATIONS OF SAM AND SAH LEVELS ARE ALSO PRODUCED IN BRAIN. METHODS: MALE ADULT SPRAGUE DAWLEY RATS WERE FED CHRONICALLY WITH LIEBER-DECARLI ETHANOL (ETOH) (9% V/V) OR CONTROL DIET. THE ETOH-DIET-FED RATS WERE WITHDRAWN FOR 0 AND 24 HOURS. THE CEREBELLUM AND LIVER TISSUES WERE DISSECTED AND USED TO INVESTIGATE CHANGES IN ONE-CARBON METABOLISM, SAM, AND SAH LEVELS. RESULTS: WE FOUND THAT CHRONIC ETOH EXPOSURE DECREASED SAM LEVELS, SAM/SAH RATIO, MS, METHYLENE TETRAHYDROFOLATE REDUCTASE, AND BETAINE HOMOCYSTEINE METHYLTRANSFERASE (BHMT) EXPRESSION AND INCREASED METHIONINE ADENOSYLTRANSFERASE-2B (MAT2B) BUT NOT MAT2A EXPRESSION IN THE LIVER. IN CONTRAST, CHRONIC ETOH EXPOSURE DECREASED SAH LEVELS, INCREASED SAM/SAH RATIO AND THE EXPRESSION OF MAT2A AND S-ADENOSYL HOMOCYSTEINE HYDROLASE, WHILE THE LEVELS OF SAM OR BHMT EXPRESSION IN CEREBELLUM REMAINED UNALTERED. HOWEVER, IN BOTH LIVER AND CEREBELLUM, CHRONIC ETOH EXPOSURE DECREASED THE EXPRESSION OF MS AND INCREASED MAT2B EXPRESSION. ALL CHRONIC ETOH-INDUCED CHANGES OF ONE-CARBON METABOLISM IN CEREBELLUM, BUT NOT LIVER, RETURNED TO NEAR-NORMAL LEVELS DURING ETOH WITHDRAWAL. CONCLUSIONS: THESE RESULTS INDICATE A DECREASED "METHYLATION INDEX" IN LIVER AND AN INCREASED "METHYLATION INDEX" IN CEREBELLUM. THE OPPOSING CHANGES OF THE "METHYLATION INDEX" SUGGEST ALTERED DNA METHYLATION IN LIVER AND CEREBELLUM, THUS IMPLICATING ONE-CARBON METABOLISM IN THE PATHOPHYSIOLOGY OF ALCOHOLISM. 2017 13 3331 40 HISTONE DEACETYLASE INHIBITOR SUBERANILOHYDROXAMIC ACID TREATMENT REVERSES HYPOSENSITIVITY TO GAMMA-AMINOBUTYRIC ACID IN THE VENTRAL TEGMENTAL AREA DURING ETHANOL WITHDRAWAL. BACKGROUND: THE VENTRAL TEGMENTAL AREA (VTA) IS IMPORTANT FOR ALCOHOL-RELATED REWARD AND REINFORCEMENT. MOUSE VTA NEURONS ARE HYPOSENSITIVE TO GAMMA-AMINOBUTYRIC ACID (GABA) DURING ETHANOL (ETOH) WITHDRAWAL, AND GABA RESPONSIVENESS IS NORMALIZED BY IN VITRO TREATMENT WITH HISTONE DEACETYLASE INHIBITORS (HDACI). THE PRESENT STUDY EXAMINED THE EFFECT OF A SYSTEMICALLY ADMINISTERED HDACI, SUBERANILOHYDROXAMIC ACID (SAHA) ON GABA SENSITIVITY, AND RELATED MOLECULAR CHANGES IN VTA NEURONS DURING WITHDRAWAL AFTER CHRONIC ETOH INTAKE IN RATS. METHODS: SPRAGUE DAWLEY MALE ADULT RATS WERE FED WITH LIEBER-DECARLI DIET (9% ETOH OR CONTROL DIET) FOR 16 DAYS. EXPERIMENTAL GROUPS INCLUDED CONTROL DIET-FED AND ETOH DIET-FED (0- OR 24-HOUR WITHDRAWAL) RATS TREATED WITH EITHER SAHA OR VEHICLE INJECTION. SINGLE-UNIT RECORDINGS WERE USED TO MEASURE THE RESPONSE OF VTA NEURONS TO GABA. IMMUNOHISTOCHEMISTRY WAS PERFORMED TO EXAMINE LEVELS OF HDAC2, ACETYLATED HISTONE H3 LYSINE 9 (ACH3K9), AND GABA(A) RECEPTOR ALPHA1 AND ALPHA5 SUBUNITS IN THE VTA; QUANTITATIVE POLYMERASE CHAIN REACTION WAS PERFORMED TO EXAMINE THE MRNA LEVELS OF HDAC2 AND GABA(A) RECEPTOR SUBUNITS. RESULTS: VTA NEURONS FROM THE WITHDRAWAL GROUP EXHIBITED GABA HYPOSENSITIVITY. IN VIVO SAHA TREATMENT 2 HOURS BEFORE SACRIFICE NORMALIZED THE SENSITIVITY OF VTA NEURONS TO GABA. ETOH WITHDRAWAL WAS ASSOCIATED WITH INCREASED HDAC2 AND DECREASED ACH3K9 PROTEIN LEVELS; SAHA TREATMENT NORMALIZED ACH3K9 LEVELS. INTERESTINGLY, NO SIGNIFICANT CHANGE WAS OBSERVED IN THE MRNA LEVELS OF HDAC2. THE MRNA LEVELS, BUT NOT PROTEIN LEVELS, OF GABA(A) RECEPTOR ALPHA1 AND ALPHA5 SUBUNITS WERE INCREASED DURING WITHDRAWAL. CONCLUSIONS: WITHDRAWAL FROM CHRONIC ETOH EXPOSURE RESULTS IN A DECREASE IN GABA-MEDIATED INHIBITION, AND THIS GABA HYPOSENSITIVITY IS NORMALIZED BY IN VIVO SAHA TREATMENT. DISRUPTION OF SIGNALING IN THE VTA PRODUCED BY ALTERATION OF GABA NEUROTRANSMISSION COULD BE 1 NEUROADAPTIVE PHYSIOLOGICAL PROCESS LEADING TO CRAVING AND RELAPSE. THESE RESULTS SUGGEST THAT HDACI PHARMACOTHERAPY WITH AGENTS LIKE SAHA MIGHT BE AN EFFECTIVE TREATMENT FOR ALCOHOLISM. 2018 14 4683 36 NEW PERSPECTIVES ON FOLATE TRANSPORT IN RELATION TO ALCOHOLISM-INDUCED FOLATE MALABSORPTION--ASSOCIATION WITH EPIGENOME STABILITY AND CANCER DEVELOPMENT. FOLATES ARE MEMBERS OF THE B-CLASS OF VITAMINS, WHICH ARE REQUIRED FOR THE SYNTHESIS OF PURINES AND PYRIMIDINES, AND FOR THE METHYLATION OF ESSENTIAL BIOLOGICAL SUBSTANCES, INCLUDING PHOSPHOLIPIDS, DNA, AND NEUROTRANSMITTERS. FOLATES CANNOT BE SYNTHESIZED DE NOVO BY MAMMALS; HENCE, AN EFFICIENT INTESTINAL ABSORPTION PROCESS IS REQUIRED. INTESTINAL FOLATE TRANSPORT IS CARRIER-MEDIATED, PH-DEPENDENT AND ELECTRONEUTRAL, WITH SIMILAR AFFINITY FOR OXIDIZED AND REDUCED FOLIC ACID DERIVATIVES. THE VARIOUS TRANSPORTERS, I.E. REDUCED FOLATE CARRIER, PROTON-COUPLED FOLATE TRANSPORTER, FOLATE-BINDING PROTEIN, AND ORGANIC ANION TRANSPORTERS, ARE INVOLVED IN THE FOLATE TRANSPORT PROCESS IN VARIOUS TISSUES. ANY IMPAIRMENT IN UPTAKE OF FOLATE CAN LEAD TO A STATE OF FOLATE DEFICIENCY, THE MOST PREVALENT VITAMIN DEFICIENCY IN WORLD, AFFECTING 10% OF THE POPULATION IN THE USA. SUCH IMPAIRMENTS IN FOLATE TRANSPORT OCCUR IN A VARIETY OF CONDITIONS, INCLUDING CHRONIC USE OF ETHANOL, SOME INBORN HEREDITARY DISORDERS, AND CERTAIN DISEASES. AMONG THESE, ETHANOL INGESTION HAS BEEN THE MAJOR CONTRIBUTOR TO FOLATE DEFICIENCY. ETHANOL-ASSOCIATED FOLATE DEFICIENCY CAN DEVELOP BECAUSE OF DIETARY INADEQUACY, INTESTINAL MALABSORPTION, ALTERED HEPATOBILIARY METABOLISM, ENHANCED COLONIC METABOLISM, AND INCREASED RENAL EXCRETION. ETHANOL REDUCES THE INTESTINAL AND RENAL UPTAKE OF FOLATE BY ALTERING THE BINDING AND TRANSPORT KINETICS OF FOLATE TRANSPORT SYSTEMS. ALSO, ETHANOL REDUCES THE EXPRESSION OF FOLATE TRANSPORTERS IN BOTH INTESTINE AND KIDNEY, AND THIS MIGHT BE A CONTRIBUTING FACTOR FOR FOLATE MALABSORPTION, LEADING TO FOLATE DEFICIENCY. THE MAINTENANCE OF INTRACELLULAR FOLATE HOMEOSTASIS IS ESSENTIAL FOR THE ONE-CARBON TRANSFER REACTIONS NECESSARY FOR DNA SYNTHESIS AND BIOLOGICAL METHYLATION REACTIONS. DNA METHYLATION IS AN IMPORTANT EPIGENETIC DETERMINANT IN GENE EXPRESSION, IN THE MAINTENANCE OF DNA INTEGRITY AND STABILITY, IN CHROMOSOMAL MODIFICATIONS, AND IN THE DEVELOPMENT OF MUTATIONS. ETHANOL, A TOXIN THAT IS CONSUMED REGULARLY, HAS BEEN FOUND TO AFFECT THE METHYLATION OF DNA. IN ADDITION TO ITS EFFECT ON DNA METHYLATION DUE TO FOLATE DEFICIENCY, ETHANOL COULD DIRECTLY EXERT ITS EFFECT THROUGH ITS INTERACTION WITH ONE-CARBON METABOLISM, IMPAIRMENT OF METHYL GROUP SYNTHESIS, AND AFFECTING THE ENZYMES REGULATING THE SYNTHESIS OF S-ADENOSYLMETHIONINE, THE PRIMARY METHYL GROUP DONOR FOR MOST BIOLOGICAL METHYLATION REACTIONS. THUS, ETHANOL PLAYS AN IMPORTANT ROLE IN THE PATHOGENESIS OF SEVERAL DISEASES THROUGH ITS POTENTIAL ABILITY TO MODULATE THE METHYLATION OF BIOLOGICAL MOLECULES. THIS REVIEW DISCUSSES THE UNDERLYING MECHANISM OF FOLATE MALABSORPTION IN ALCOHOLISM, THE MECHANISM OF METHYLATION-ASSOCIATED SILENCING OF GENES, AND HOW THE INTERACTION BETWEEN ETHANOL AND FOLATE DEFICIENCY AFFECTS THE METHYLATION OF GENES, THEREBY MODULATING EPIGENOME STABILITY AND THE RISK OF CANCER. 2009 15 2156 39 EPIGENETIC MECHANISMS ARE INVOLVED IN THE REGULATION OF ETHANOL CONSUMPTION IN MICE. BACKGROUND: REPEATED ALCOHOL EXPOSURE IS KNOWN TO INCREASE SUBSEQUENT ETHANOL CONSUMPTION IN MICE. HOWEVER, THE UNDERLYING MECHANISMS HAVE NOT BEEN FULLY ELUCIDATED. ONE POSTULATED MECHANISM INVOLVES EPIGENETIC MODIFICATIONS, INCLUDING HISTONE MODIFICATIONS AND DNA METHYLATION OF RELEVANT GENES SUCH AS NR2B OR BDNF. METHODS: TO INVESTIGATE THE ROLE OF EPIGENETIC MECHANISMS IN THE DEVELOPMENT OF ALCOHOL DRINKING BEHAVIOR, AN ESTABLISHED CHRONIC INTERMITTENT ETHANOL EXPOSURE REINFORCED ETHANOL DRINKING MOUSE MODEL WITH VAPOR INHALATION OVER TWO 9-DAY TREATMENT REGIMENS WAS USED. THE DNA METHYLTRANSFERASE INHIBITOR, 5-AZACYTIDINE OR THE HISTONE DEACETYLASE INHIBITOR, TRICHOSTATIN A WAS ADMINISTERED (INTRAPERITONEALLY) TO C57BL/6 MICE 30 MIN BEFORE DAILY EXPOSURE TO CHRONIC INTERMITTENT ETHANOL. CHANGES IN ETHANOL CONSUMPTION WERE MEASURED USING THE 2-BOTTLE CHOICE TEST. RESULTS: THE RESULTS INDICATED THAT SYSTEMIC ADMINISTRATION OF TRICHOSTATIN A (2.5 MICROG/G) FACILITATED CHRONIC INTERMITTENT ETHANOL-INDUCED ETHANOL DRINKING, BUT SYSTEMIC ADMINISTRATION OF 5-AZACYTIDINE (2 MICROG/G) DID NOT CAUSE THE SAME EFFECT. HOWEVER, WHEN 5-AZACYTIDINE WAS ADMINISTERED BY INTRACEREBROVENTRICULAR INJECTION, IT FACILITATED CHRONIC INTERMITTENT ETHANOL-INDUCED ETHANOL DRINKING. FURTHERMORE, THE INCREASED DRINKING CAUSED BY CHRONIC INTERMITTENT ETHANOL WAS PREVENTED BY INJECTION OF A METHYL DONOR, S-ADENOSYL-L-METHIONINE. TO PROVIDE EVIDENCE THAT CHRONIC INTERMITTENT ETHANOL- OR TRICHOSTATIN A-INDUCED DNA DEMETHYLATION AND HISTONE MODIFICATIONS OF THE NR2B PROMOTER MAY UNDERLIE THE ALTERED ETHANOL CONSUMPTION, WE EXAMINED EPIGENETIC MODIFICATIONS AND NR2B EXPRESSION IN THE PREFRONTAL CORTEX OF THESE MICE. CHRONIC INTERMITTENT ETHANOL OR TRICHOSTATIN A DECREASED DNA METHYLATION AND INCREASED HISTONE ACETYLATION IN THE NR2B GENE PROMOTER, AS WELL AS MRNA LEVELS OF NR2B IN THESE MICE. CONCLUSIONS: TAKEN TOGETHER, THESE RESULTS INDICATE THAT EPIGENETIC MODIFICATIONS ARE INVOLVED IN REGULATING ETHANOL DRINKING BEHAVIOR, PARTIALLY THROUGH ALTERING NR2B EXPRESSION. 2014 16 6445 28 THERAPEUTIC BENEFITS OF THE METHYL DONOR S-ADENOSYLMETHIONINE ON NERVE INJURY-INDUCED MECHANICAL HYPERSENSITIVITY AND COGNITIVE IMPAIRMENT IN MICE. DESPITE CONSIDERABLE ADVANCES IN UNDERSTANDING MECHANISMS INVOLVED IN CHRONIC PAIN, EFFECTIVE TREATMENT REMAINS ELUSIVE. COMORBID CONDITIONS INCLUDING ANXIETY, DEPRESSION, AND COGNITIVE IMPAIRMENT FURTHER IMPACT QUALITY OF LIFE. CHRONIC PAIN IS ASSOCIATED WITH REVERSIBLE CHANGES IN BRAIN ANATOMY AND FUNCTION AND WITH LONG-TERM CHANGES IN GENE EXPRESSION. EPIGENETIC MECHANISMS, INCLUDING DNA METHYLATION, CONTRIBUTE TO WIDE-SPREAD AND LONG-LASTING REPROGRAMMING OF GENE EXPRESSION. WE PREVIOUSLY REPORTED DECREASES IN GLOBAL DNA METHYLATION IN THE MOUSE FRONTAL CORTEX 6 MONTHS AFTER INDUCTION OF NEUROPATHIC PAIN USING THE SPARED NERVE INJURY (SNI) MODEL. HERE, WE EXAMINED THE THERAPEUTIC EFFECT OF INCREASING DNA METHYLATION USING THE METHYL DONOR S-ADENOSYLMETHIONINE (SAM). S-ADENOSYLMETHIONINE IS MARKETED AS A NUTRITIONAL SUPPLEMENT FOR A RANGE OF CONDITIONS INCLUDING LIVER DISEASE, DEPRESSION, OSTEOARTHRITIS, FIBROMYALGIA, AND DEMENTIA. THREE MONTHS AFTER SNI OR SHAM SURGERY, ANIMALS WERE TREATED WITH SAM (20 MG/KG, 3X/WEEK) OR SALINE ORALLY FOR 4 MONTHS, AND THE IMPACT ON SENSORY, MOTOR, MOTIVATIONAL, AND COGNITIVE INDICES WAS MEASURED. S-ADENOSYLMETHIONINE ATTENUATED SNI-INDUCED MECHANICAL HYPERSENSITIVITY AND REDUCED ACTIVE AVOIDANCE OF MECHANICAL STIMULI BUT HAD NO EFFECT ON COLD SENSITIVITY OR MOTOR CAPACITY. S-ADENOSYLMETHIONINE COMPLETELY BLOCKED NERVE INJURY-INDUCED COGNITIVE IMPAIRMENT AND ATTENUATED SNI-INDUCED DECREASES IN GLOBAL DNA METHYLATION IN THE FRONTAL CORTEX. IN SUMMARY, CHRONIC ORAL ADMINISTRATION OF THE METHYL DONOR, SAM, ATTENUATED SENSORY AND COGNITIVE SYMPTOMS ASSOCIATED WITH NERVE INJURY IN MICE. THESE EFFECTS MAY BE MEDIATED, IN PART, THROUGH MODULATION OF DNA METHYLATION IN THE CENTRAL NERVOUS SYSTEM BY SYSTEMIC ADMINISTRATION OF THE METHYL DONOR SAM. 2017 17 3727 34 INHIBITION OF PANCREATIC ACINAR MITOCHONDRIAL THIAMIN PYROPHOSPHATE UPTAKE BY THE CIGARETTE SMOKE COMPONENT 4-(METHYLNITROSAMINO)-1-(3-PYRIDYL)-1-BUTANONE. THIAMIN IS ESSENTIAL FOR NORMAL METABOLISM IN PANCREATIC ACINAR CELLS (PAC) AND IS OBTAINED FROM THEIR MICROENVIRONMENT THROUGH SPECIFIC PLASMA-MEMBRANE TRANSPORTERS, CONVERTED TO THIAMIN PYROPHOSPHATE (TPP) IN THE CYTOPLASM, FOLLOWED BY UPTAKE OF TPP BY MITOCHONDRIA THROUGH THE MITOCHONDRIAL TPP (MTPP) TRANSPORTER (MTPPT; PRODUCT OF SLC25A19 GENE). TPP IS ESSENTIAL FOR NORMAL MITOCHONDRIAL FUNCTION. WE EXAMINED THE EFFECT OF LONG-TERM/CHRONIC EXPOSURE OF PAC IN VITRO (PANCREATIC ACINAR 266-6 CELLS) AND IN VIVO (WILD-TYPE OR TRANSGENIC MICE CARRYING THE SLC25A19 PROMOTER) OF THE CIGARETTE SMOKE TOXIN, 4-(METHYLNITROSAMINO)-1-(3-PYRIDYL)-1-BUTANONE (NNK), ON THE MTPP UPTAKE PROCESS. OUR IN VITRO AND IN VIVO FINDINGS DEMONSTRATE THAT NNK NEGATIVELY AFFECTS MTPP UPTAKE AND REDUCED EXPRESSION OF MTPPT PROTEIN, MTPPT MRNA, AND HETEROGENOUS NUCLEAR RNA, AS WELL AS SLC25A19 PROMOTER ACTIVITY. THE EFFECT OF NNK ON SLC25A19 TRANSCRIPTION WAS NEITHER MEDIATED BY CHANGES IN EXPRESSION OF TRANSCRIPTIONAL FACTOR NFY-1 (KNOWN TO DRIVE SLC25A19 TRANSCRIPTION), NOR DUE TO CHANGES IN METHYLATION PROFILE OF THE SLC25A19 PROMOTER. RATHER, IT APPEARS TO BE DUE TO CHANGES IN HISTONE MODIFICATIONS THAT INVOLVE SIGNIFICANT DECREASES IN HISTONE H3K4-TRIMETHYLATION AND H3K9-ACETYLATION (ACTIVATION MARKERS). THE EFFECT OF NNK ON MTPPT FUNCTION IS MEDIATED THROUGH THE NONNEURONAL ALPHA7-NICOTINIC ACETYLCHOLINE RECEPTOR (ALPHA7-NACHR), AS INDICATED BY BOTH IN VITRO (USING THE NACHR ANTAGONIST MECAMYLAMINE) AND IN VIVO (USING AN ALPHA7-NACHR(-/-) MOUSE MODEL) STUDIES. THESE FINDINGS DEMONSTRATE THAT CHRONIC EXPOSURE OF PAC TO NNK NEGATIVELY IMPACTS PAC MTPP UPTAKE. THIS EFFECT APPEARS TO BE EXERTED AT THE LEVEL OF SLC25A19 TRANSCRIPTION, INVOLVE EPIGENETIC MECHANISM(S), AND IS MEDIATED THROUGH THE ALPHA7-NACHR. 2016 18 893 38 CHRONIC ETHANOL EXPOSURE AND WITHDRAWAL IMPAIR SYNAPTIC GABA(A) RECEPTOR-MEDIATED NEUROTRANSMISSION IN DEEP-LAYER PREFRONTAL CORTEX. BACKGROUND: THE PREFRONTAL CORTEX (PFC) ACTS AS AN INTEGRATIVE HUB FOR THE PROCESSING OF CORTICAL AND SUBCORTICAL INPUT INTO MEANINGFUL EFFERENT SIGNALING, PERMITTING COMPLEX ASSOCIATIVE BEHAVIORS. PFC DYSFUNCTION IS CONSISTENTLY OBSERVED WITH ETHANOL (ETOH) DEPENDENCE AND IS A CORE COMPONENT OF THE PATHOLOGY OF ALCOHOL USE DISORDERS IN CURRENT MODELS OF ADDICTION. WHILE INTRACORTICAL GAMMA-AMINOBUTRYRIC ACID (GABA)ERGIC NEUROTRANSMISSION IS UNDERSTOOD TO BE ESSENTIAL FOR MAINTAINING COORDINATED NETWORK ACTIVITY WITHIN THE CORTEX, RELATIVELY LITTLE IS KNOWN REGARDING FUNCTIONAL GABAERGIC ADAPTATIONS IN PFC DURING ETOH DEPENDENCE. METHODS: IN THE PRESENT STUDY, MALE AND FEMALE (> POSTNATAL DAY 60) SPRAGUE-DAWLEY RATS WERE ADMINISTERED ETOH (5.0 G/KG; INTRAGASTRIC GAVAGE) FOR 14 TO 15 CONSECUTIVE DAYS. TWENTY-FOUR HOURS AFTER THE FINAL ADMINISTRATION, ANIMALS WERE SACRIFICED AND BRAINS EXTRACTED FOR ELECTROPHYSIOLOGICAL RECORDINGS OF ISOLATED GABA(A) RECEPTOR-MEDIATED CURRENTS OR ANALYSIS OF GABA(A) RECEPTOR SUBUNIT PROTEIN EXPRESSION IN DEEP-LAYER PFC NEURONS. RESULTS: CHRONIC ETOH EXPOSURE SIGNIFICANTLY ATTENUATED ACTIVITY-DEPENDENT SPONTANEOUS GABA(A) RECEPTOR-MEDIATED INHIBITORY POSTSYNAPTIC CURRENT (IPSC) FREQUENCY WITH NO EFFECT ON AMPLITUDE. FURTHERMORE, ANALYSIS OF IPSC DECAY KINETICS REVEALED A SIGNIFICANT ENHANCEMENT OF IPSC DECAY TIME THAT WAS ASSOCIATED WITH DECREMENTS IN EXPRESSION OF THE ALPHA1 GABA(A) RECEPTOR SUBUNIT, INDICATIVE OF FURTHER IMPAIRED PHASIC INHIBITION. THESE PHENOMENA OCCURRED IRRESPECTIVE OF NEURON PROJECTION DESTINATION AND SEX. BASED ON PREVIOUS OBSERVATIONS BY OUR LABORATORY OF AN EPIGENETIC MECHANISM FOR ETOH-INDUCED CHANGES IN CORTICAL GABA(A) RECEPTOR SUBUNIT EXPRESSION, THE HISTONE DEACETYLASE INHIBITOR TRICHOSTATIN A WAS ADMINISTERED TO WATER- AND ETOH-EXPOSED ANIMALS, AND PREVENTED ETOH-INDUCED CHANGES IN SPONTANEOUS IPSC FREQUENCY, IPSC DECAY KINETICS, AND GABA(A) RECEPTOR SUBUNIT EXPRESSION. CONCLUSIONS: TAKEN TOGETHER, THESE RESULTS DEMONSTRATE THAT CHRONIC ETOH EXPOSURE IMPAIRS SYNAPTIC INHIBITORY NEUROTRANSMISSION IN DEEP-LAYER PYRAMIDAL NEURONS OF THE MEDIAL PFC IN BOTH MALE AND FEMALE RATS. THESE MALADAPTATIONS OCCUR IN NEURONS PROJECTING TO NUMEROUS REGIONS IMPLICATED IN THE SEQUELAE OF ETOH DEPENDENCE, OFFERING A MECHANISTIC LINK BETWEEN THE MANIFESTATION OF PFC DYSFUNCTION AND NEGATIVE AFFECTIVE STATES OBSERVED WITH EXTENDED CONSUMPTION. 2019 19 4819 20 OCCURRENCE OF TOXICITY AND CELL PROLIFERATION AFTER A SINGLE GAVAGE ADMINISTRATION OF CHLOROFORM TO MALE F344 RATS. CHLOROFORM, AN INDUSTRIAL SOLVENT AND ONE OF THE MOST COMMON ENVIRONMENTAL CONTAMINANTS WHICH PRODUCES CARCINOGENIC EFFECTS IN THE LIVER AND KIDNEY OF RODENTS, IS NOT GENOTOXIC IN MOST TRADITIONAL BACTERIAL AND MAMMALIAN TEST SYSTEMS. ITS CARCINOGENIC POTENTIAL APPEARS ATTRIBUTABLE TO THE SUSTAINED CELL TURNOVER (REGENERATIVE HYPERPLASIA) WHICH RESULTS FROM CHRONIC CHLOROFORM TOXICITY. IN THIS PRESENT STUDY, CELL PROLIFERATION (REPLICATIVE DNA SYNTHESIS, RDS) AND HISTOPATHOLOGICAL CHANGES IN HEPATOCYTES AND RENAL TUBULAR EPITHELIAL CELLS WERE ASSESSED IN MALE F344 RATS FOLLOWING A SINGLE GAVAGE CHLOROFORM EXPOSURE (50, 150 OR 500 MG/KG). IN ADDITION, BIOCHEMICAL PARAMETERS (BUN, GOT, LDH AND NAG) WERE EXAMINED USING PLASMA AND URINE SAMPLES. CELL PROLIFERATION AND HISTOPATHOLOGICAL CHANGES (E.G. HYPERTROPHY, NECROSIS, VACUOLATION) WERE ONLY SEEN AT THE DOSE OF 500 MG/KG IN THE LIVER AND KIDNEY. AT THE SAME DOSE, ALL BIOCHEMICAL MARKERS WERE INCREASED AT THE 24 TO 48 HR TIME POINTS. THESE RESULTS OBTAINED ARE THUS IN LINE WITH EARLIER FINDINGS POINTING TO EPIGENETIC CARCINOGENICITY. 1998 20 3295 43 HIGH PHOSPHATE-INDUCED DOWNREGULATION OF PPARGAMMA CONTRIBUTES TO CKD-ASSOCIATED VASCULAR CALCIFICATION. MEDIAL ARTERIAL CALCIFICATION ASSOCIATED WITH HYPERPHOSPHATEMIA IS A MAIN CAUSE OF CARDIOVASCULAR MORTALITY IN PATIENTS WITH CHRONIC KIDNEY DISEASE (CKD), BUT THE MECHANISMS UNDERLYING HIGH PHOSPHATE-INDUCED VASCULAR CALCIFICATION REMAIN LARGELY UNKNOWN. HERE, WE OBSERVED A SIGNIFICANT DECREASE IN THE EXPRESSION OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-GAMMA (PPARGAMMA) IN CALCIFIED ARTERIES BOTH IN CKD PATIENTS AND IN A MOUSE MODEL OF CKD WITH HYPERPHOSPHATEMIA. IN VITRO, HIGH PHOSPHATE TREATMENT LED TO A DECREASED EXPRESSION OF PPARGAMMA IN MOUSE VASCULAR SMOOTH MUSCLE CELLS (VMSCS), ACCOMPANIED BY APPARENT OSTEOGENIC DIFFERENTIATION AND CALCIFICATION. PRETREATMENT WITH PPARGAMMA AGONIST ROSIGLITAZONE SIGNIFICANTLY REVERSED HIGH PHOSPHATE-INDUCED VSMCS CALCIFICATION. FURTHER INVESTIGATION SHOWED THAT METHYL-CPG BINDING PROTEIN 2 (MECP2)-MEDIATED EPIGENETIC REPRESSION WAS INVOLVED IN HIGH PHOSPHATE-INDUCED PPARGAMMA DOWNREGULATION. MOREOVER, THE EXPRESSION OF KLOTHO THAT HAS THE ABILITY TO INHIBIT VASCULAR CALCIFICATION BY REGULATING PHOSPHATE UPTAKE DECREASED WITH THE PPARGAMMA REDUCTION IN VSMCS AFTER HIGH PHOSPHATE TREATMENT, AND ROSIGLITAZONE FAILED TO INHIBIT HIGH PHOSPHATE-INDUCED CALCIFICATION IN VSMCS WITH KNOCKDOWN OF KLOTHO OR IN AORTIC RINGS FROM KLOTHO-DEFICIENT (KL/KL) MICE. FINALLY, AN IN VIVO STUDY DEMONSTRATED THAT ORAL ADMINISTRATION OF ROSIGLITAZONE COULD INCREASE KLOTHO EXPRESSION AND PROTECT AGAINST HIGH PHOSPHATE-INDUCED VASCULAR CALCIFICATION IN CKD MICE. THESE FINDINGS SUGGEST THAT THE INHIBITION OF PPARGAMMA EXPRESSION MAY CONTRIBUTE TO THE PATHOGENESIS OF HIGH PHOSPHATE-INDUCED VASCULAR CALCIFICATION, WHICH MAY PROVIDE A NEW THERAPEUTIC TARGET FOR VASCULAR CALCIFICATION IN CKD PATIENTS. 2018