1  4275 132 MICROGLIA ACTIVATION IN THE MIDBRAIN OF THE HUMAN NEONATE: THE EFFECT OF PERINATAL HYPOXIC-ISCHEMIC INJURY. PERINATAL HYPOXIA-ISCHEMIA (PHI) IS A MAJOR RISK FACTOR FOR THE DEVELOPMENT OF NEUROPSYCHIATRIC DEFICITS LATER IN LIFE. WE PREVIOUSLY REPORTED THAT AFTER PROLONGED PHI, THE DOPAMINERGIC NEURONS OF THE HUMAN NEONATE SHOWED A DRAMATIC REDUCTION OF TYROSINE HYDROXYLASE (TH) IN THE SUBSTANTIA NIGRA, WITHOUT IMPORTANT SIGNS OF NEURONAL DEGENERATION DESPITE THE SIGNIFICANT REDUCTION IN THEIR CELL SIZE. SINCE MICROGLIA ACTIVATION COULD PRECEDE NEURONAL DEATH, WE NOW INVESTIGATED 2 MICROGLIA ACTIVATION MARKERS, IONIZED CALCIUM-BINDING ADAPTER MOLECULE 1 (IBA1), AND THE PHAGOCYTOSIS MARKER CD68. THE HIGHEST IBA1 IMMUNOREACTIVITY WAS FOUND IN NEONATES WITH NEUROPATHOLOGICAL LESIONS OF SEVERE/ABRUPT PHI, WHILE THE LOWEST IN SUBJECTS WITH MODERATE/PROLONGED OR OLDER PHI. SUBJECTS WITH VERY SEVERE/PROLONGED OR CHRONIC PHI SHOWED AN INCREASED IBA1 EXPRESSION AND VERY ACTIVATED MICROGLIAL MORPHOLOGY. HEAVY ATTACHMENT OF MICROGLIA ON TH NEURONS AND REMARKABLE EXPRESSION OF CD68 WERE ALSO OBSERVED INDICATING PHAGOCYTOSIS IN THIS GROUP. FEMALES APPEAR TO EXPRESS MORE IBA1 THAN MALES, SUGGESTING A GENDER DIFFERENCE IN MICROGLIA MATURATION AND IMMUNE REACTIVITY AFTER PHI INSULT. PHI-INDUCED MICROGLIAL "PRIMING" DURING THE SENSITIVE FOR BRAIN DEVELOPMENT PERINATAL/NEONATAL PERIOD, IN COMBINATION WITH GENETIC OR OTHER EPIGENETIC FACTORS, COULD PREDISPOSE THE SURVIVORS TO NEUROPSYCHIATRIC DISORDERS LATER IN LIFE, POSSIBLY THROUGH A SEXUALLY DIMORPHIC WAY.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
2  1619  38 DNA METHYLTRANSFERASE REGULATES NITRIC OXIDE HOMEOSTASIS AND VIRULENCE IN A CHRONICALLY ADAPTED PSEUDOMONAS AERUGINOSA STRAIN. OPPORTUNISTIC PATHOGENS SUCH AS PSEUDOMONAS AERUGINOSA ADAPT THEIR GENOMES RAPIDLY DURING CHRONIC INFECTIONS. UNDERSTANDING THEIR EPIGENETIC REGULATION MAY PROVIDE BIOMARKERS FOR DIAGNOSIS AND REVEAL NOVEL REGULATORY MECHANISMS. WE PERFORMED SINGLE-MOLECULE REAL-TIME SEQUENCING (SMRT-SEQ) TO CHARACTERIZE THE METHYLOME OF A CHRONICALLY ADAPTED P. AERUGINOSA CLINICAL STRAIN, TBCF10839. TWO N(6)-METHYLADENINE (6MA) METHYLATION RECOGNITION MOTIFS (RCCANNNNNNNTGAR AND TRGANNNNNNTGC [MODIFICATION SITES ARE IN BOLD]) WERE IDENTIFIED AND PREDICTED AS NEW TYPE I METHYLATION SITES USING REBASE ANALYSIS. WE CONFIRMED THAT THE MOTIF TRGANNNNNNTGC WAS METHYLATED BY THE METHYLTRANSFERASE (MTASE) M.PAETBCFII, ACCORDING TO METHYLATION SENSITIVITY ASSAYS IN VIVO AND VITRO. TRANSCRIPTOMIC ANALYSIS SHOWED THAT A DELTAPAETBCFIIM KNOCKOUT MUTANT SIGNIFICANTLY DOWNREGULATED NITRIC OXIDE REDUCTASE (NOR) REGULATION AND EXPRESSION OF CODING GENES SUCH AS NOSR AND NORB, WHICH CONTAIN METHYLATED MOTIFS IN THEIR PROMOTERS OR CODING REGIONS. THE DELTAPAETBCFIIM STRAIN EXHIBITED REDUCED INTERCELLULAR SURVIVAL CAPACITY IN NO-PRODUCING RAW264.7 MACROPHAGES AND ATTENUATED VIRULENCE IN A GALLERIA MELLONELLA INFECTION MODEL; THE COMPLEMENTED STRAIN RECOVERED THESE DEFECTIVE PHENOTYPES. FURTHER PHYLOGENETIC ANALYSIS DEMONSTRATED THAT HOMOLOGS OF M.PAETBCFII OCCUR FREQUENTLY IN P. AERUGINOSA AS WELL AS OTHER BACTERIAL SPECIES. OUR WORK THEREFORE PROVIDED NEW INSIGHTS INTO THE RELATIONSHIP BETWEEN DNA METHYLATION, NO DETOXIFICATION, AND BACTERIAL VIRULENCE, LAYING A FOUNDATION FOR FURTHER EXPLORING THE MOLECULAR MECHANISM OF DNA METHYLTRANSFERASE IN REGULATING THE PATHOGENICITY OF P. AERUGINOSA. IMPORTANCE PSEUDOMONAS AERUGINOSA IS AN OPPORTUNISTIC PATHOGEN WHICH CAUSES ACUTE AND CHRONIC INFECTIONS THAT ARE DIFFICULT TO TREAT. COMPARATIVE GENOMIC ANALYSIS HAS SHOWED BROAD GENOME DIVERSITY AMONG P. AERUGINOSA CLINICAL STRAINS AND REVEALED THEIR DIFFERENT REGULATORY TRAITS COMPARED TO THE LABORATORY STRAINS. WHILE CURRENT INVESTIGATION OF THE EPIGENETICS OF P. AERUGINOSA IS STILL LACKING, UNDERSTANDING EPIGENETIC REGULATION MAY PROVIDE BIOMARKERS FOR DIAGNOSIS AND FACILITATE DEVELOPMENT OF NOVEL THERAPIES. DENITRIFICATION CAPABILITY IS CRITICAL FOR MICROBIAL VERSATILITY IN RESPONSE TO DIFFERENT ENVIRONMENTAL STRESS CONDITIONS, INCLUDING THE BACTERIAL INFECTION PROCESS, WHERE NITRIC OXIDE (NO) CAN BE GENERATED BY PHAGOCYTIC CELLS. THE DENITRIFICATION REGULATION MECHANISMS HAVE BEEN STUDIED INTENSIVELY AT GENETIC AND BIOCHEMICAL LEVELS. HOWEVER, THERE IS VERY LITTLE EVIDENCE ABOUT THE EPIGENETIC REGULATION OF BACTERIAL DENITRIFICATION MECHANISM. P. AERUGINOSA TBCF10839 IS A CHRONICALLY HOST-ADAPTED STRAIN ISOLATED FROM A CYSTIC FIBROSIS (CF) PATIENT WITH SPECIAL ANTIPHAGOCYTOSIS CHARACTERISTICS. HERE, WE INVESTIGATED THE REGULATORY EFFECT OF AN ORPHAN DNA MTASE, M.PAETBCFII, IN P. AERUGINOSA TBCF10839. WE DEMONSTRATED THAT THE DNA MTASE REGULATES THE TRANSCRIPTION OF DENITRIFICATION GENES REPRESENTED BY NOR AND AFFECTS ANTIPHAGOCYTIC ABILITY IN BACTERIA. IN SILICO ANALYSIS SUGGESTED THAT DNA METHYLATION MODIFICATION MAY ENHANCE GENE EXPRESSION BY AFFECTING THE BINDING OF TRANSACTING FACTORS SUCH AS DNR AND RPON. OUR FINDINGS NOT ONLY DEEPEN THE UNDERSTANDING OF THE ROLE OF DNA MTASE IN TRANSCRIPTIONAL REGULATION IN P. AERUGINOSA BUT ALSO PROVIDE A THEORETICAL FOUNDATION FOR THE IN-DEPTH STUDY OF THE MOLECULAR MECHANISM OF THE EPIGENETIC REGULATION ON DENITRIFICATION, VIRULENCE, AND HOST-PATHOGEN INTERACTION.	2022	

3  2300  29 EPIGENETIC REGULATION OF BDNF EXPRESSION IN THE PRIMARY SENSORY NEURONS AFTER PERIPHERAL NERVE INJURY: IMPLICATIONS IN THE DEVELOPMENT OF NEUROPATHIC PAIN. BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) IS KNOWN TO BE UP-REGULATED IN THE DORSAL ROOT GANGLION (DRG) AFTER PERIPHERAL NERVE INJURY, AND TO CONTRIBUTE TO NEUROPATHIC PAIN. HERE, WE FOUND THAT THERMAL HYPERALGESIA AND MECHANICAL ALLODYNIA AT DAY 7 POST-INJURY WERE INHIBITED ONLY WHEN ANTI-BDNF ANTIBODY WAS INTRATHECALLY ADMINISTRATED AT DAY 2 POST-INJURY. CONSISTENT WITH BEHAVIORAL RESULTS, WESTERN BLOT ANALYSIS SHOWED THAT THE EXPRESSION LEVELS OF BDNF PROTEIN IN THE SPINAL DORSAL HORN WERE MARKEDLY INDUCED DURING EARLY STAGE POST-INJURY. MOREOVER, THE MAXIMAL INCREASE IN BDNF MRNA EXPRESSION IN THE DRG WAS OBSERVED AT DAY 1 POST-INJURY, AND SIGNIFICANTLY ELEVATED LEVELS WERE SUSTAINED FOR AT LEAST 14 DAYS. FOUR OF FIVE BDNF MRNA TRANSCRIPTS WERE UP-REGULATED AFTER NERVE INJURY, AND THE MOST INDUCIBLE TRANSCRIPT WAS EXON I. USING A CHROMATIN IMMUNOPRECIPITATION (CHIP) ASSAY, WE FOUND THAT NERVE INJURY PROMOTES HISTONE H3 AND H4 ACETYLATION, TRANSCRIPTIONALLY ACTIVE MODIFICATIONS, AT BDNF PROMOTER I AT DAY 1 POST-INJURY, AND THE LEVELS OF HISTONE ACETYLATION REMAIN ELEVATED FOR AT LEAST 7 DAYS. TAKEN TOGETHER, OUR FINDINGS SUGGEST THAT AN INITIAL INCREASE IN BDNF EXON I EXPRESSION CONTROLLED BY EPIGENETIC MECHANISMS MIGHT HAVE A CRUCIAL ROLE IN THE DEVELOPMENT OF NEUROPATHIC PAIN.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
4  5823  26 STRESS INCREASES DNA METHYLATION OF THE NEURONAL PAS DOMAIN 4 (NPAS4) GENE. NEURONAL PER ARNT SIM DOMAIN 4 (NPAS4), A BRAIN-SPECIFIC HELIX-LOOP-HELIX TRANSCRIPTION FACTOR, WAS RECENTLY SHOWN TO REGULATE THE DEVELOPMENT OF GABAERGIC INHIBITORY NEURONS. NPAS4 MRNA EXPRESSION LEVELS WERE DECREASED IN THE HIPPOCAMPUS OF MICE EXPOSED TO STRESS, WHICH WAS ACCOMPANIED BY BRAIN DYSFUNCTION. WE HAVE SUGGESTED THAT TRANSIENT STRESS REDUCED NPAS4 TRANSCRIPTION THROUGH THE GLUCOCORTICOID RECEPTOR. IN THE PRESENT REPORT, WE INVESTIGATED THE POTENTIAL CONTRIBUTION OF EPIGENETIC MODIFICATIONS INDUCED BY STRESS ON NPAS4 GENE EXPRESSION. THE NPAS4 PROMOTER REGION CONTAINS TWO CPG ISLANDS; IN THE HIPPOCAMPUS, CHRONIC RESTRAINT STRESS INCREASES THE DNA METHYLATION LEVELS OF BOTH OF THESE CPG ISLANDS. IN THE NEURO2A CELL LINE, TREATMENT WITH A DNA METHYLTRANSFERASE INHIBITOR, 5-AZA-2'-DEOXYCYTIDINE, INCREASED NPAS4 MRNA LEVELS AND MARKEDLY REDUCED THE DNA METHYLATION LEVELS OF CPG ISLAND 2 IN THE NPAS4 PROMOTER. THE DNA METHYLATION SITES IN CPG ISLAND 2 OVERLAP WITH TWO CYCLIC ADENOSINE MONOPHOSPHATE RESPONSE ELEMENT (CRE) SEQUENCES. MUTATION OF THESE CRE SEQUENCES REDUCED NPAS4 PROMOTER ACTIVITY. THESE RESULTS SUGGEST THAT TRANSCRIPTION OF THE NPAS4 GENE IS DOWNREGULATED BY STRESS THROUGH DNA METHYLATION OF ITS PROMOTER.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
5  4699  26 NFATC2-DEPENDENT EPIGENETIC UPREGULATION OF CXCL14 IS INVOLVED IN THE DEVELOPMENT OF NEUROPATHIC PAIN INDUCED BY PACLITAXEL. BACKGROUND: THE MAJOR DOSE-LIMITING TOXICITY OF PACLITAXEL, ONE OF THE MOST COMMONLY USED DRUGS TO TREAT SOLID TUMOR, IS PAINFUL NEUROPATHY. HOWEVER, THE MOLECULAR MECHANISMS UNDERLYING PACLITAXEL-INDUCED PAINFUL NEUROPATHY ARE LARGELY UNCLARIFIED. METHODS: PAW WITHDRAWAL THRESHOLD WAS MEASURED IN THE RATS FOLLOWING INTRAPERITONEAL INJECTION OF PACLITAXEL. THE QPCR, WESTERN BLOTTING, PROTEIN OR CHROMATIN IMMUNOPRECIPITATION, CHIP-SEQ IDENTIFICATION OF NFATC2 BINDING SITES, AND MICROARRAY ANALYSIS WERE PERFORMED TO EXPLORE THE MOLECULAR MECHANISM. RESULTS: WE FOUND THAT PACLITAXEL TREATMENT INCREASED THE NUCLEAR EXPRESSION OF NFATC2 IN THE SPINAL DORSAL HORN, AND KNOCKDOWN OF NFATC2 WITH NFATC2 SIRNA SIGNIFICANTLY ATTENUATED THE MECHANICAL ALLODYNIA INDUCED BY PACLITAXEL. FURTHER BINDING SITE ANALYSIS UTILIZING CHIP-SEQ ASSAY COMBINING WITH GENE EXPRESSION PROFILE REVEALED A SHIFT OF NFATC2 BINDING SITE CLOSER TO TTS OF TARGET GENES IN DORSAL HORN AFTER PACLITAXEL TREATMENT. WE FURTHER FOUND THAT NFATC2 OCCUPANCY MAY DIRECTLY UPREGULATE THE CHEMOKINE CXCL14 EXPRESSION IN DORSAL HORN, WHICH WAS MEDIATED BY ENHANCED INTERACTION BETWEEN NFATC2 AND P300 AND CONSEQUENTLY INCREASED ACETYLATION OF HISTONE H4 IN CXCL14 PROMOTER REGION. ALSO, KNOCKDOWN OF CXCL14 IN DORSAL HORN SIGNIFICANTLY ATTENUATED MECHANICAL ALLODYNIA INDUCED BY PACLITAXEL. CONCLUSION: THESE RESULTS SUGGESTED THAT ENHANCED INTERACTION BETWEEN P300 AND NFATC2 MEDIATED THE EPIGENETIC UPREGULATION OF CXCL14 IN THE SPINAL DORSAL HORN, WHICH CONTRIBUTED TO THE CHEMOTHERAPEUTIC PACLITAXEL-INDUCED CHRONIC PAIN.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
6  3625  35 IN VIVO GENOME AND METHYLOME ADAPTATION OF CAG-NEGATIVE HELICOBACTER PYLORI DURING EXPERIMENTAL HUMAN INFECTION. MULTIPLE STUDIES HAVE DEMONSTRATED RAPID BACTERIAL GENOME EVOLUTION DURING CHRONIC INFECTION WITH HELICOBACTER PYLORI IN CONTRAST, LITTLE WAS KNOWN ABOUT GENETIC CHANGES DURING THE FIRST STAGES OF INFECTION, WHEN SELECTIVE PRESSURE IS LIKELY TO BE HIGHEST. USING SINGLE-MOLECULE, REAL-TIME (SMRT) AND ILLUMINA SEQUENCING TECHNOLOGIES, WE ANALYZED GENOME AND METHYLOME EVOLUTION DURING THE FIRST 10 WEEKS OF INFECTION BY COMPARING THE CAG PATHOGENICITY ISLAND (CAGPAI)-NEGATIVE H. PYLORI CHALLENGE STRAIN BCS 100 WITH PAIRS OF H. PYLORI REISOLATES FROM GASTRIC ANTRUM AND CORPUS BIOPSY SPECIMENS OF 10 HUMAN VOLUNTEERS WHO HAD BEEN INFECTED WITH THIS STRAIN AS PART OF A VACCINE TRIAL. MOST GENETIC CHANGES DETECTED IN THE REISOLATES AFFECTED GENES WITH A SURFACE-RELATED ROLE OR A PREDICTED FUNCTION IN PEPTIDE UPTAKE. APART FROM PHENOTYPIC CHANGES OF THE BACTERIAL ENVELOPE, A DUPLICATION OF THE CATALASE GENE WAS OBSERVED IN ONE REISOLATE, WHICH RESULTED IN HIGHER CATALASE ACTIVITY AND IMPROVED SURVIVAL UNDER OXIDATIVE STRESS CONDITIONS. THE METHYLOMES ALSO VARIED IN SOME OF THE REISOLATES, MOSTLY BY ACTIVITY SWITCHING OF PHASE-VARIABLE METHYLTRANSFERASE (MTASE) GENES. THE OBSERVED IN VIVO MUTATION SPECTRUM WAS REMARKABLE FOR A VERY HIGH PROPORTION OF NONSYNONYMOUS MUTATIONS. ALTHOUGH THE DATA SHOWED SUBSTANTIAL WITHIN-STRAIN GENOME DIVERSITY IN THE CHALLENGE STRAIN, MOST ANTRUM AND CORPUS REISOLATES FROM THE SAME VOLUNTEERS WERE HIGHLY SIMILAR TO EACH OTHER, INDICATING THAT THE CHALLENGE INFECTION REPRESENTS A MAJOR SELECTIVE BOTTLENECK SHAPING THE TRANSMITTED POPULATION. OUR FINDINGS SUGGEST RAPID IN VIVO SELECTION OF H. PYLORI DURING EARLY-PHASE INFECTION PROVIDING ADAPTATION TO DIFFERENT INDIVIDUALS BY COMMON MECHANISMS OF GENETIC AND EPIGENETIC ALTERATIONS.IMPORTANCE EXCEPTIONAL GENETIC DIVERSITY AND VARIABILITY ARE HALLMARKS OF HELICOBACTER PYLORI, BUT THE BIOLOGICAL ROLE OF THIS PLASTICITY REMAINS INCOMPLETELY UNDERSTOOD. HERE, WE HAD THE RARE OPPORTUNITY TO INVESTIGATE THE MOLECULAR EVOLUTION DURING THE FIRST WEEKS OF H. PYLORI INFECTION BY COMPARING THE GENOMES AND EPIGENOMES OF H. PYLORI STRAIN BCS 100 USED TO CHALLENGE HUMAN VOLUNTEERS IN A VACCINE TRIAL WITH THOSE OF BACTERIA REISOLATED FROM THE VOLUNTEERS 10 WEEKS AFTER THE CHALLENGE. THE DATA PROVIDE MOLECULAR INSIGHTS INTO THE PROCESS OF ESTABLISHMENT OF THIS HIGHLY VERSATILE PATHOGEN IN 10 DIFFERENT HUMAN INDIVIDUAL HOSTS, SHOWING, FOR EXAMPLE, SELECTION FOR CHANGES IN HOST-INTERACTION MOLECULES AS WELL AS CHANGES IN EPIGENETIC METHYLATION PATTERNS. THE DATA PROVIDE IMPORTANT CLUES TO THE EARLY ADAPTATION OF H. PYLORI TO NEW HOST NICHES AFTER TRANSMISSION, WHICH WE BELIEVE IS VITAL TO UNDERSTAND ITS SUCCESS AS A CHRONIC PATHOGEN AND DEVELOP MORE EFFICIENT TREATMENTS AND VACCINES.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
7   949  25 CHRONIC METHAMPHETAMINE TREATMENT REDUCES THE EXPRESSION OF SYNAPTIC PLASTICITY GENES AND CHANGES THEIR DNA METHYLATION STATUS IN THE MOUSE BRAIN. METHAMPHETAMINE (METH) IS A HIGHLY ADDICTIVE PSYCHOSTIMULANT THAT MAY CAUSE LONG-LASTING SYNAPTIC DYSFUNCTION AND ABNORMAL GENE EXPRESSION. WE AIMED TO EXPLORE THE DIFFERENTIAL EXPRESSION OF SYNAPTIC PLASTICITY GENES IN CHRONIC METH-TREATED MOUSE BRAIN. WE USED THE RT(2) PROFILER PCR ARRAY AND THE REAL-TIME QUANTITATIVE PCR TO CHARACTERIZE DIFFERENTIALLY EXPRESSED SYNAPTIC PLASTICITY GENES IN THE FRONTAL CORTEX AND THE HIPPOCAMPUS OF CHRONIC METH-TREATED MICE COMPARED WITH NORMAL SALINE-TREATED MICE. WE FURTHER USED PYROSEQUENCING TO ASSESS DNA METHYLATION CHANGES IN THE CPG REGION OF THE FIVE IMMEDIATE EARLY GENES (IEGS) IN CHRONIC METH-TREATED MOUSE BRAIN. WE DETECTED SIX DOWNREGULATED GENES IN THE FRONTAL CORTEX AND THE HIPPOCAMPUS OF CHRONIC METH-TREATED MICE, INCLUDING FIVE IEGS (ARC, EGR2, FOS, KLF10, AND NR4A1) AND ONE NEURONAL RECEPTOR GENE (GRM1), COMPARED WITH NORMAL SALINE-TREATED GROUP, BUT ONLY FOUR GENES (ARC, EGR2, FOS, AND NR4A1) WERE CONFIRMED TO BE DIFFERENT. FURTHERMORE, WE FOUND SEVERAL CPG SITES OF THE ARC AND THE FOS THAT HAD SIGNIFICANT CHANGES IN DNA METHYLATION STATUS IN THE FRONTAL CORTEX OF CHRONIC METH-TREATED MICE, WHILE THE KLF10 AND THE NR4A1 THAT HAD SIGNIFICANT CHANGES IN THE HIPPOCAMPUS. OUR RESULTS SHOW THAT CHRONIC ADMINISTRATION OF METH MAY LEAD TO SIGNIFICANT DOWNREGULATION OF THE IEGS EXPRESSION IN BOTH THE FRONTAL CORTEX AND THE HIPPOCAMPUS, WHICH MAY PARTLY ACCOUNT FOR THE MOLECULAR MECHANISM OF THE ACTION OF METH. FURTHERMORE, THE CHANGES IN DNA METHYLATION STATUS OF THE IEGS IN THE BRAIN INDICATE THAT AN EPIGENETIC MECHANISM-DEPENDENT TRANSCRIPTIONAL REGULATION MAY CONTRIBUTE TO METH ADDICTION, WHICH WARRANTS ADDITIONAL STUDY.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
8  5760  28 SOLUBLE URIC ACID PRIMES TLR-INDUCED PROINFLAMMATORY CYTOKINE PRODUCTION BY HUMAN PRIMARY CELLS VIA INHIBITION OF IL-1RA. OBJECTIVES: THE STUDY OF THE PROINFLAMMATORY ROLE OF URIC ACID HAS FOCUSED ON THE EFFECTS OF ITS CRYSTALS OF MONOSODIUM URATE (MSU). HOWEVER, LITTLE IS KNOWN WHETHER URIC ACID ITSELF CAN DIRECTLY HAVE PROINFLAMMATORY EFFECTS. IN THIS STUDY, WE INVESTIGATE THE PRIMING EFFECTS OF URIC ACID EXPOSURE ON THE CYTOKINE PRODUCTION OF PRIMARY HUMAN CELLS UPON STIMULATION WITH GOUT-RELATED STIMULI. METHODS: PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) WERE HARVESTED FROM PATIENTS WITH GOUT AND HEALTHY VOLUNTEERS. CELLS WERE PRETREATED WITH OR WITHOUT URIC ACID IN SOLUBLE FORM FOR 24 H AND THEN STIMULATED FOR 24 H WITH TOLL-LIKE RECEPTOR (TLR)2 OR TLR4 LIGANDS IN THE PRESENCE OR ABSENCE OF MSU CRYSTALS. CYTOKINE PRODUCTION WAS MEASURED BY ELISA; MRNA LEVELS WERE ASSESSED USING QPCR. RESULTS: THE PRODUCTION OF INTERLEUKIN (IL)-1BETA AND IL-6 WAS HIGHER IN PATIENTS COMPARED WITH CONTROLS AND THIS CORRELATED WITH SERUM URATE LEVELS. PROINFLAMMATORY CYTOKINE PRODUCTION WAS SIGNIFICANTLY POTENTIATED WHEN CELLS FROM HEALTHY SUBJECTS WERE PRETREATED WITH URIC ACID. SURPRISINGLY, THIS WAS ASSOCIATED WITH A SIGNIFICANT DOWNREGULATION OF THE ANTI-INFLAMMATORY CYTOKINE IL-1 RECEPTOR ANTAGONIST (IL-1RA). THIS EFFECT WAS SPECIFIC TO STIMULATION BY URIC ACID AND WAS EXERTED AT THE LEVEL OF GENE TRANSCRIPTION. EPIGENETIC REPROGRAMMING AT THE LEVEL OF HISTONE METHYLATION BY URIC ACID WAS INVOLVED IN THIS EFFECT. CONCLUSIONS: IN THIS STUDY WE DEMONSTRATE A MECHANISM THROUGH WHICH HIGH CONCENTRATIONS OF URIC ACID (UP TO 50 MG/DL) INFLUENCE INFLAMMATORY RESPONSES BY FACILITATING IL-1BETA PRODUCTION IN PBMCS. WE SHOW THAT A MECHANISM FOR THE AMPLIFICATION OF IL-1BETA CONSISTS IN THE DOWNREGULATION OF IL-1RA AND THAT THIS EFFECT COULD BE EXERTED VIA EPIGENETIC MECHANISMS SUCH AS HISTONE METHYLATION. HYPERURICAEMIA CAUSES A SHIFT IN THE IL-1BETA/IL-1RA BALANCE PRODUCED BY PBMCS AFTER EXPOSURE TO MSU CRYSTALS AND TLR-MEDIATED STIMULI, AND THIS PHENOMENON IS LIKELY TO REINFORCE THE ENHANCED STATE OF CHRONIC INFLAMMATION.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
9  3201  24 HDAC2 IN PRIMARY SENSORY NEURONS CONSTITUTIVELY RESTRAINS CHRONIC PAIN BY REPRESSING ALPHA2DELTA-1 EXPRESSION AND ASSOCIATED NMDA RECEPTOR ACTIVITY. ALPHA2DELTA-1 (ENCODED BY THE CACNA2D1 GENE) IS A NEWLY DISCOVERED NMDA RECEPTOR-INTERACTING PROTEIN AND IS THE THERAPEUTIC TARGET OF GABAPENTINOIDS (E.G., GABAPENTIN AND PREGABALIN) FREQUENTLY USED FOR TREATING PATIENTS WITH NEUROPATHIC PAIN. NERVE INJURY CAUSES SUSTAINED ALPHA2DELTA-1 UPREGULATION IN THE DORSAL ROOT GANGLION (DRG), WHICH PROMOTES NMDA RECEPTOR SYNAPTIC TRAFFICKING AND ACTIVATION IN THE SPINAL DORSAL HORN, A HALLMARK OF CHRONIC NEUROPATHIC PAIN. HOWEVER, LITTLE IS KNOWN ABOUT HOW NERVE INJURY INITIATES AND MAINTAINS THE HIGH EXPRESSION LEVEL OF ALPHA2DELTA-1 TO SUSTAIN CHRONIC PAIN. HERE, WE SHOW THAT NERVE INJURY CAUSED HISTONE HYPERACETYLATION AND DIMINISHED ENRICHMENT OF HISTONE DEACETYLASE-2 (HDAC2), BUT NOT HDAC3, AT THE CACNA2D1 PROMOTER IN THE DRG. STRIKINGLY, HDAC2 KNOCKDOWN OR CONDITIONAL KNOCKOUT IN DRG NEURONS IN MALE AND FEMALE MICE CONSISTENTLY INDUCED LONG-LASTING MECHANICAL PAIN HYPERSENSITIVITY, WHICH WAS READILY REVERSED BY BLOCKING NMDA RECEPTORS, INHIBITING ALPHA2DELTA-1 WITH GABAPENTIN OR DISRUPTING THE ALPHA2DELTA-1-NMDA RECEPTOR INTERACTION AT THE SPINAL CORD LEVEL. HDAC2 DELETION IN DRG NEURONS INCREASED HISTONE ACETYLATION LEVELS AT THE CACNA2D1 PROMOTER, UPREGULATED ALPHA2DELTA-1 IN THE DRG, AND POTENTIATED ALPHA2DELTA-1-DEPENDENT NMDA RECEPTOR ACTIVITY AT PRIMARY AFFERENT CENTRAL TERMINALS IN THE SPINAL DORSAL HORN. CORRESPONDINGLY, HDAC2 KNOCKDOWN-INDUCED PAIN HYPERSENSITIVITY WAS BLUNTED IN CACNA2D1 KNOCKOUT MICE. THUS, OUR FINDINGS REVEAL THAT HDAC2 FUNCTIONS AS A PIVOTAL TRANSCRIPTIONAL REPRESSOR OF NEUROPATHIC PAIN VIA CONSTITUTIVELY SUPPRESSING ALPHA2DELTA-1 EXPRESSION AND ENSUING PRESYNAPTIC NMDA RECEPTOR ACTIVITY IN THE SPINAL CORD. HDAC2 ENRICHMENT LEVELS AT THE CACNA2D1 PROMOTER IN DRG NEURONS CONSTITUTE A UNIQUE EPIGENETIC MECHANISM THAT GOVERNS ACUTE-TO-CHRONIC PAIN TRANSITION.SIGNIFICANCE STATEMENT EXCESS ALPHA2DELTA-1 PROTEINS PRODUCED AFTER NERVE INJURY DIRECTLY INTERACT WITH GLUTAMATE NMDA RECEPTORS TO POTENTIATE SYNAPTIC NMDA RECEPTOR ACTIVITY IN THE SPINAL CORD, A PROMINENT MECHANISM OF NERVE PAIN. BECAUSE ALPHA2DELTA-1 UPREGULATION AFTER NERVE INJURY IS LONG LASTING, GABAPENTINOIDS RELIEVE PAIN SYMPTOMS ONLY TEMPORARILY. OUR STUDY DEMONSTRATES FOR THE FIRST TIME THE UNEXPECTED ROLE OF INTRINSIC HDAC2 ACTIVITY AT THE ALPHA2DELTA-1 GENE PROMOTER IN LIMITING ALPHA2DELTA-1 GENE TRANSCRIPTION, NMDA RECEPTOR-DEPENDENT SYNAPTIC PLASTICITY, AND CHRONIC PAIN DEVELOPMENT AFTER NERVE INJURY. THESE FINDINGS CHALLENGE THE PREVAILING VIEW ABOUT THE ROLE OF GENERAL HDAC ACTIVITY IN PROMOTING CHRONIC PAIN. RESTORING THE REPRESSIVE HDAC2 FUNCTION AND/OR REDUCING HISTONE ACETYLATION AT THE ALPHA2DELTA-1 GENE PROMOTER IN PRIMARY SENSORY NEURONS COULD LEAD TO LONG-LASTING RELIEF OF NERVE PAIN.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
10  3037  30 GENOME AND METHYLOME VARIATION IN HELICOBACTER PYLORI WITH A CAG PATHOGENICITY ISLAND DURING EARLY STAGES OF HUMAN INFECTION. BACKGROUND & AIMS: HELICOBACTER PYLORI IS REMARKABLE FOR ITS GENETIC VARIATION; YET, LITTLE IS KNOWN ABOUT ITS GENETIC CHANGES DURING EARLY STAGES OF HUMAN INFECTION, AS THE BACTERIA ADAPT TO THEIR NEW ENVIRONMENT. WE ANALYZED GENOME AND METHYLOME VARIATIONS IN A FULLY VIRULENT STRAIN OF H PYLORI DURING EXPERIMENTAL INFECTION. METHODS: WE PERFORMED A RANDOMIZED PHASE I/II, OBSERVER-BLIND, PLACEBO-CONTROLLED STUDY OF 12 HEALTHY, H PYLORI-NEGATIVE ADULTS IN GERMANY FROM OCTOBER 2008 THROUGH MARCH 2010. THE VOLUNTEERS WERE GIVEN A PROPHYLACTIC VACCINE CANDIDATE (N = 7) OR PLACEBO (N = 5) AND THEN CHALLENGED WITH H PYLORI STRAIN BCM-300. BIOPSY SAMPLES WERE COLLECTED AND H PYLORI WERE ISOLATED. GENOMES OF THE CHALLENGE STRAIN AND 12 REISOLATES, OBTAINED 12 WEEKS AFTER (OR IN 1 CASE, 62 WEEKS AFTER) INFECTION WERE SEQUENCED BY SINGLE-MOLECULE, REAL-TIME TECHNOLOGY, WHICH, IN PARALLEL, PERMITTED DETERMINATION OF GENOME-WIDE METHYLATION PATTERNS FOR ALL STRAINS. FUNCTIONAL EFFECTS OF GENETIC CHANGES OBSERVED IN H PYLORI STRAINS DURING HUMAN INFECTION WERE ASSESSED BY MEASURING RELEASE OF INTERLEUKIN 8 FROM AGS CELLS (TO DETECT CAG PATHOGENICITY ISLAND FUNCTION), NEUTRAL RED UPTAKE (TO DETECT VACUOLATING CYTOTOXIN ACTIVITY), AND ADHESION ASSAYS. RESULTS: THE OBSERVED MUTATION RATE WAS IN AGREEMENT WITH RATES PREVIOUSLY DETERMINED FROM PATIENTS WITH CHRONIC H PYLORI INFECTIONS, WITHOUT EVIDENCE OF A MUTATION BURST. A LOSS OF CAG PATHOGENICITY ISLAND FUNCTION WAS OBSERVED IN 3 REISOLATES. IN ADDITION, 3 REISOLATES FROM THE VACCINE GROUP ACQUIRED MUTATIONS IN THE VACUOLATING CYTOTOXIN GENE VACA, RESULTING IN LOSS OF VACUOLIZATION ACTIVITY. WE OBSERVED INTERSTRAIN VARIATION IN METHYLOMES DUE TO PHASE VARIATION IN GENES ENCODING METHYLTRANSFERASES. CONCLUSIONS: WE ANALYZED ADAPTATION OF A FULLY VIRULENT STRAIN OF H PYLORI TO 12 DIFFERENT VOLUNTEERS TO OBTAIN A ROBUST ESTIMATE OF THE FREQUENCY OF GENETIC AND EPIGENETIC CHANGES IN THE ABSENCE OF INTERSTRAIN RECOMBINATION. OUR FINDINGS INDICATE THAT THE LARGE AMOUNT OF GENETIC VARIATION IN H PYLORI POSES A CHALLENGE TO VACCINE DEVELOPMENT. CLINICALTRIALS.GOV NO: NCT00736476.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
11   579  28 BEHAVIORAL AND MOLECULAR NEUROEPIGENETIC ALTERATIONS IN PRENATALLY STRESSED MICE: RELEVANCE FOR THE STUDY OF CHROMATIN REMODELING PROPERTIES OF ANTIPSYCHOTIC DRUGS. WE HAVE RECENTLY REPORTED THAT MICE BORN FROM DAMS STRESSED DURING PREGNANCY (PRS MICE), IN ADULTHOOD, HAVE BEHAVIORAL DEFICITS REMINISCENT OF BEHAVIORS OBSERVED IN SCHIZOPHRENIA (SZ) AND BIPOLAR (BP) DISORDER PATIENTS. FURTHERMORE, WE HAVE SHOWN THAT THE FRONTAL CORTEX (FC) AND HIPPOCAMPUS OF ADULT PRS MICE, LIKE THAT OF POSTMORTEM CHRONIC SZ PATIENTS, ARE CHARACTERIZED BY INCREASES IN DNA-METHYLTRANSFERASE 1 (DNMT1), TEN-ELEVEN METHYLCYTOSINE DIOXYGENASE 1 (TET1) AND EXHIBIT AN ENRICHMENT OF 5-METHYLCYTOSINE (5MC) AND 5-HYDROXYMETHYLCYTOSINE (5HMC) AT NEOCORTICAL GABAERGIC AND GLUTAMATERGIC GENE PROMOTERS. HERE, WE SHOW THAT THE BEHAVIORAL DEFICITS AND THE INCREASED 5MC AND 5HMC AT GLUTAMIC ACID DECARBOXYLASE 67 (GAD1), REELIN (RELN) AND BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) PROMOTERS AND THE REDUCED EXPRESSION OF THE MESSENGER RNAS (MRNAS) AND PROTEINS CORRESPONDING TO THESE GENES IN FC OF ADULT PRS MICE IS REVERSED BY TREATMENT WITH CLOZAPINE (5 MG KG(-1) TWICE A DAY FOR 5 DAYS) BUT NOT BY HALOPERIDOL (1 MG KG(-1) TWICE A DAY FOR 5 DAYS). INTERESTINGLY, CLOZAPINE HAD NO EFFECT ON EITHER THE BEHAVIOR, PROMOTER METHYLATION OR THE EXPRESSION OF THESE MRNAS AND PROTEINS WHEN ADMINISTERED TO OFFSPRING OF NONSTRESSED PREGNANT MICE. CLOZAPINE, BUT NOT HALOPERIDOL, REDUCED THE ELEVATED LEVELS OF DNMT1 AND TET1, AS WELL AS THE ELEVATED LEVELS OF DNMT1 BINDING TO GAD1, RELN AND BDNF PROMOTERS IN PRS MICE SUGGESTING THAT CLOZAPINE, UNLIKE HALOPERIDOL, MAY LIMIT DNA METHYLATION BY INTERFERING WITH DNA METHYLATION DYNAMICS. WE CONCLUDE THAT THE PRS MOUSE MODEL MAY BE USEFUL PRECLINICALLY IN SCREENING FOR THE POTENTIAL EFFICACY OF ANTIPSYCHOTIC DRUGS ACTING ON ALTERED EPIGENETIC MECHANISMS. FURTHERMORE, PRS MICE MAY BE INVALUABLE FOR UNDERSTANDING THE ETIOPATHOGENESIS OF SZ AND BP DISORDER AND FOR PREDICTING TREATMENT RESPONSES AT EARLY STAGES OF THE ILLNESS ALLOWING FOR EARLY DETECTION AND REMEDIAL INTERVENTION.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
12  2405  31 EPIGENETIC RESPONSE IN MICE MASTITIS: ROLE OF HISTONE H3 ACETYLATION AND MICRORNA(S) IN THE REGULATION OF HOST INFLAMMATORY GENE EXPRESSION DURING STAPHYLOCOCCUS AUREUS INFECTION. BACKGROUND: THERE IS RENEWED INTEREST TOWARDS UNDERSTANDING THE HOST-PATHOGEN INTERACTION IN THE LIGHT OF EPIGENETIC MODIFICATIONS. ALTHOUGH EPITHELIAL TISSUE IS THE MAJOR SITE FOR HOST-PATHOGEN INTERACTIONS, THERE IS HANDFUL OF STUDIES TO SHOW HOW EPITHELIAL CELLS RESPOND TO PATHOGENS. BACTERIAL INFECTION IN THE MAMMARY GLAND PARENCHYMA INDUCES LOCAL AND SUBSEQUENTLY SYSTEMIC INFLAMMATION THAT RESULTS IN A COMPLEX DISEASE CALLED MASTITIS. GLOBALLY STAPHYLOCOCCUS AUREUS IS THE SINGLE LARGEST MASTITIS PATHOGEN AND THE INFECTION CAN ULTIMATELY RESULT IN EITHER SUBCLINICAL OR CHRONIC AND SOMETIMES LIFELONG INFECTION. RESULTS: IN THE PRESENT REPORT WE HAVE ADDRESSED THE DIFFERENTIAL INFLAMMATORY RESPONSE IN MICE MAMMARY TISSUE DURING INTRAMAMMARY INFECTION AND THE ALTERED EPIGENETIC CONTEXT INDUCED BY TWO CLOSELY RELATED STRAINS OF S. AUREUS, ISOLATED FROM FIELD SAMPLES. IMMUNOHISTOCHEMICAL AND IMMUNOBLOTTING ANALYSIS SHOWED STRAIN SPECIFIC HYPERACETYLATION AT HISTONE H3K9 AND H3K14 RESIDUES. GLOBAL GENE EXPRESSION ANALYSIS IN S. AUREUS INFECTED MICE MAMMARY TISSUE REVEALED A SELECTIVE SET OF UPREGULATED GENES THAT SIGNIFICANTLY CORRELATED WITH THE PROMOTER SPECIFIC, HISTONE H3K14 ACETYLATION. FURTHERMORE, WE HAVE IDENTIFIED SEVERAL DIFFERENTIALLY EXPRESSED KNOWN MIRNAS AND 3 NOVEL MIRNAS IN S. AUREUS INFECTED MICE MAMMARY TISSUE BY SMALL RNA SEQUENCING. BY EMPLOYING THESE GENE EXPRESSION DATA, AN ATTEMPT HAS BEEN MADE TO DELINEATE THE GENE REGULATORY NETWORKS IN THE STRAIN SPECIFIC INFLAMMATORY RESPONSE. APPARENTLY, ONE OF THE ISOLATES OF S. AUREUS ACTIVATED THE NF-KAPPAB SIGNALING LEADING TO DRASTIC INFLAMMATORY RESPONSE AND INDUCTION OF IMMUNE SURVEILLANCE, WHICH COULD POSSIBLY LEAD TO RAPID CLEARANCE OF THE PATHOGEN. THE OTHER STRAIN REPRESSED MOST OF THE INFLAMMATORY RESPONSE, WHICH MIGHT HELP IN ITS SUSTENANCE IN THE HOST TISSUE. CONCLUSION: TAKEN TOGETHER, OUR STUDIES SHED SUBSTANTIAL LIGHTS TO UNDERSTAND THE MECHANISMS OF STRAIN SPECIFIC DIFFERENTIAL INFLAMMATORY RESPONSE TO S. AUREUS INFECTION DURING MASTITIS. IN A BROADER PERSPECTIVE THIS STUDY ALSO PAVES THE WAY TO UNDERSTAND HOW CERTAIN BACTERIA CAN EVADE THE IMMUNE SURVEILLANCE AND CAUSE SUSTAINED INFECTION WHILE OTHERS ARE RAPIDLY CLEARED FROM THE HOST BODY.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
13  4491  31 MONOSOMY 7 MYELOPROLIFERATIVE DISEASE IN CHILDREN WITH NEUROFIBROMATOSIS, TYPE 1: EPIDEMIOLOGY AND MOLECULAR ANALYSIS. LOSS OF CONSTITUTIONAL HETEROZYGOSITY IS A COMMON MOLECULAR FEATURE OF CANCERS IN WHICH INACTIVATION OF ONE OR MORE TUMOR SUPPRESSOR GENES IS THOUGHT TO CONTRIBUTE TO TUMORIGENESIS. RECENT EVIDENCE SUGGESTS THAT THE GENE RESPONSIBLE FOR NEUROFIBROMATOSIS, TYPE 1 (NF-1), BELONGS TO THIS CLASS OF HERITABLE CANCER GENES. CHILDREN WITH NF-1 SHOW AN INCREASED INCIDENCE OF MYELOID LEUKEMIA, INCLUDING JUVENILE CHRONIC MYELOGENOUS LEUKEMIA (JCML) AND, PERHAPS, THE MYELOPROLIFERATIVE SYNDROME (MPS) ASSOCIATED WITH BONE MARROW MONOSOMY 7 (MO 7). WE HAVE INVESTIGATED FIVE CHILDREN WITH MO 7: THREE WITH NF-1 AND TWO OTHERS WITH SUGGESTIVE EVIDENCE OF NF-1. SOUTHERN BLOTTING EXPERIMENTS PERFORMED IN FOUR PATIENTS SHOWED NO LOSS OF HETEROZYGOSITY IN BONE MARROW SPECIMENS USING PROBES LINKED TO THE NF-1 LOCUS ON THE LONG ARM OF CHROMOSOME 17. BOTH OF OUR PATIENTS WITH FAMILIAL NF-1 INHERITED THE DISEASE FROM THEIR MOTHERS, AS DID 14 OF 19 OTHER CASES OF MYELOID LEUKEMIA IN CHILDREN WITH FAMILIAL NF-1. SEVENTEEN OF THESE 21 CHILDREN WERE BOYS. MYELOID LEUKEMIA DEVELOPED IN 12 BOYS AND FOUR GIRLS WHO INHERITED NF-1 FROM THEIR MOTHERS, AND IN FIVE BOYS WHO INHERITED THE DISEASE FROM THEIR FATHERS. FATHER-TO-DAUGHTER TRANSMISSION WAS NOT OBSERVED. TAKEN TOGETHER, THE PRESENCE OF CHROMOSOME 7 DELETIONS IN THE LEUKEMIAS OF CHILDREN WITH NF-1, A PATTERN OF INHERITANCE FAVORING MATERNAL TRANSMISSION OF NF-1, AND THE MARKED PREDILECTION FOR BOYS TO DEVELOP JCML AND MO 7 SUGGEST A MULTISTEP MECHANISM OF ONCOGENESIS IN WHICH EPIGENETIC FACTORS MIGHT PLAY A ROLE. FURTHER INVESTIGATION IS REQUIRED TO DETERMINE IF THE NF-1 GENES IN THE LEUKEMIC BONE MARROWS OF THESE PATIENTS HAVE ACQUIRED POINT MUTATIONS OR SMALL DELETIONS.	1992	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
14   219  27 ACUTE IMMOBILIZATION STRESS FOLLOWING CONTEXTUAL FEAR CONDITIONING REDUCES FEAR MEMORY: TIMING IS ESSENTIAL. BACKGROUND: HISTONE ACETYLATION IS REGULATED IN RESPONSE TO STRESS AND PLAYS AN IMPORTANT ROLE IN LEARNING AND MEMORY. CHRONIC STRESS IS KNOWN TO DETERIORATE COGNITION, WHEREAS ACUTE STRESS FACILITATES MEMORY FORMATION. HOWEVER, WHETHER ACUTE STRESS FACILITATES MEMORY FORMATION WHEN IT IS APPLIED AFTER FEAR STIMULATION IS NOT YET KNOWN. THEREFORE, THIS STUDY AIMED TO INVESTIGATE THE EFFECT OF ACUTE STRESS APPLIED AFTER FEAR TRAINING ON MEMORY FORMATION, MRNA EXPRESSION OF BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF), EPIGENETIC REGULATION OF BDNF EXPRESSION, AND CORTICOSTERONE LEVEL IN MICE IN VIVO. METHODS: MICE WERE SUBJECTED TO ACUTE IMMOBILIZATION STRESS FOR 30 MIN AT 60 OR 90 MIN AFTER CONTEXTUAL FEAR CONDITIONING TRAINING, AND ACETYLATION OF HISTONE 3 AT LYSINE 14 (H3K14) AND LEVEL OF CORTICOSTERONE WERE MEASURED USING WESTERN BLOT ANALYSIS AND ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA), RESPECTIVELY. A FREEZING BEHAVIOR TEST WAS PERFORMED 24 H AFTER TRAINING, AND MRNA EXPRESSION OF BDNF WAS MEASURED USING REAL-TIME POLYMERASE CHAIN REACTIONS. DIFFERENT GROUPS OF MICE WERE USED FOR EACH TEST. RESULTS: FREEZING BEHAVIOR SIGNIFICANTLY DECREASED WITH THE DOWN-REGULATION OF BDNF MRNA EXPRESSION CAUSED BY ACUTE IMMOBILIZATION STRESS AT 60 MIN AFTER FEAR CONDITIONING TRAINING OWING TO THE REDUCTION OF H3K14 ACETYLATION. HOWEVER, BDNF MRNA EXPRESSION AND H3K14 ACETYLATION WERE NOT REDUCED IN ANIMALS SUBJECTED TO IMMOBILIZATION STRESS AT 90 MIN AFTER THE TRAINING. FURTHER, THE CORTICOSTERONE LEVEL WAS SIGNIFICANTLY HIGH IN MICE SUBJECTED TO IMMOBILIZATION STRESS AT 60 MIN AFTER THE TRAINING. CONCLUSION: ACUTE IMMOBILIZATION STRESS FOR 30 MIN AT 60 MIN AFTER FEAR CONDITIONING TRAINING IMPAIRED MEMORY FORMATION AND REDUCED BDNF MRNA EXPRESSION AND H3K14 ACETYLATION IN THE HIPPOCAMPUS OF MICE OWING TO THE HIGH LEVEL OF CORTICOSTERONE.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
15  1320  25 DEMETHYLATION REGULATION OF BDNF GENE EXPRESSION IN DORSAL ROOT GANGLION NEURONS IS IMPLICATED IN OPIOID-INDUCED PAIN HYPERSENSITIVITY IN RATS. REPEATED ADMINISTRATION OF MORPHINE MAY RESULT IN OPIOID-INDUCED HYPERSENSITIVITY (OIH), WHICH INVOLVES ALTERED EXPRESSION OF NUMEROUS GENES, INCLUDING BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) IN DORSAL ROOT GANGLION (DRG) NEURONS. YET, IT REMAINS UNCLEAR HOW BDNF EXPRESSION IS INCREASED IN DRG NEURONS AFTER REPEATED MORPHINE TREATMENT. DNA METHYLATION IS AN IMPORTANT MECHANISM OF EPIGENETIC CONTROL OF GENE EXPRESSION. IN THE CURRENT STUDY, WE HYPOTHESIZED THAT THE DEMETHYLATION REGULATION OF CERTAIN BDNF GENE PROMOTERS IN DRG NEURONS MAY CONTRIBUTE TO THE DEVELOPMENT OF OIH. REAL-TIME RT-PCR WAS USED TO ASSESS CHANGES IN THE MRNA TRANSCRIPTION LEVELS OF MAJOR BDNF EXONS INCLUDING EXON I, II, IV, VI, AS WELL AS TOTAL BDNF MRNA IN DRGS FROM RATS AFTER REPEATED MORPHINE ADMINISTRATION. THE LEVELS OF EXON IV AND TOTAL BDNF MRNA WERE SIGNIFICANTLY UPREGULATED BY REPEATED MORPHINE ADMINISTRATION, AS COMPARED TO THAT IN SALINE CONTROL GROUP. FURTHER, ELISA ARRAY AND IMMUNOCYTOCHEMISTRY STUDY REVEALED A ROBUST UPREGULATION OF BDNF PROTEIN EXPRESSION IN DRG NEURONS AFTER REPEATED MORPHINE EXPOSURE. CORRESPONDINGLY, THE METHYLATION LEVELS OF BDNF EXON IV PROMOTER SHOWED A SIGNIFICANT DOWNREGULATION BY MORPHINE TREATMENT. IMPORTANTLY, INTRATHECAL ADMINISTRATION OF A BDNF ANTIBODY, BUT NOT CONTROL IGG, SIGNIFICANTLY INHIBITED MECHANICAL HYPERSENSITIVITY THAT DEVELOPED IN RATS AFTER REPEATED MORPHINE TREATMENT. CONVERSELY, INTRATHECAL ADMINISTRATION OF AN INHIBITOR OF DNA METHYLATION, 5-AZA-2'-DEOXYCYTIDINE (5-AZA-DC) MARKEDLY UPREGULATED THE BDNF PROTEIN EXPRESSION IN DRG NEURONS AND ENHANCED THE MECHANICAL ALLODYNIA AFTER REPEATED MORPHINE EXPOSURE. TOGETHER, OUR FINDINGS SUGGEST THAT DEMETHYLATION REGULATION OF BDNF GENE PROMOTER MAY BE IMPLICATED IN THE DEVELOPMENT OF OIH THROUGH EPIGENETIC CONTROL OF BDNF EXPRESSION IN DRG NEURONS.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
16  1653  32 DOPAMINE TRANSPORTER KNOCKOUT RATS DISPLAY EPIGENETIC ALTERATIONS IN RESPONSE TO COCAINE EXPOSURE. (1) BACKGROUND: THERE IS AN URGENT NEED FOR EFFECTIVE TREATMENTS FOR COCAINE USE DISORDER (CUD), AND NEW PHARMACOLOGICAL APPROACHES TARGETING EPIGENETIC MECHANISMS APPEAR TO BE PROMISING OPTIONS FOR THE TREATMENT OF THIS DISEASE. DOPAMINE TRANSPORTER (DAT) TRANSGENIC RATS RECENTLY HAVE BEEN PROPOSED AS A NEW ANIMAL MODEL FOR STUDYING SUSCEPTIBILITY TO CUD. (2) METHODS: DAT TRANSGENIC RATS WERE TREATED CHRONICALLY WITH COCAINE (10 MG/KG) FOR 8 DAYS, AND THE EXPRESSION OF EPIGENETIC MODULATORS, LYSINE DEMETHYLASE 6B (KDM6B) AND BROMODOMAIN-CONTAINING PROTEIN 4 (BRD4), WAS EXAMINED IN THE PREFRONTAL CORTEX (PFC). (3) RESULTS: WE SHOW THAT ONLY FULL KNOCKOUT (KO) OF DAT IMPACTS BASAL LEVELS OF KDM6B IN FEMALES. ADDITIONALLY, COCAINE ALTERED THE EXPRESSION OF BOTH EPIGENETIC MARKERS IN A SEX- AND GENOTYPE-DEPENDENT MANNER. IN RESPONSE TO CHRONIC COCAINE, KDM6B EXPRESSION WAS DECREASED IN MALE RATS WITH PARTIAL DAT MUTATION (HET), WHILE NO CHANGES WERE OBSERVED IN WILD-TYPE (WT) OR KO RATS. INDEED, WHILE HET MALE RATS HAVE REDUCED KDM6B AND BRD4 EXPRESSION, HET FEMALE RATS SHOWED INCREASED KDM6B AND BRD4 EXPRESSION LEVELS, HIGHLIGHTING THE IMPACT OF SEX ON EPIGENETIC MECHANISMS IN RESPONSE TO COCAINE. FINALLY, BOTH MALE AND FEMALE KO RATS SHOWED INCREASED EXPRESSION OF BRD4, BUT ONLY KO FEMALES EXHIBITED SIGNIFICANTLY INCREASED KDM6B EXPRESSION IN RESPONSE TO COCAINE. ADDITIONALLY, THE MAGNITUDE OF THESE EFFECTS WAS BIGGER IN FEMALES WHEN COMPARED TO MALES FOR BOTH EPIGENETIC ENZYMES. (4) CONCLUSIONS: THIS PRELIMINARY STUDY PROVIDES ADDITIONAL SUPPORT THAT TARGETING KDM6B AND/OR BRD4 MAY POTENTIALLY BE THERAPEUTIC IN TREATING ADDICTION-RELATED BEHAVIORS IN A SEX-DEPENDENT MANNER.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
17   718  30 CALCIUM-DEPENDENT INTRACELLULAR SIGNAL PATHWAYS IN PRIMARY CULTURED ADIPOCYTES AND ANK3 GENE VARIATION IN PATIENTS WITH BIPOLAR DISORDER AND HEALTHY CONTROLS. BIPOLAR DISORDER (BD) IS A CHRONIC PSYCHIATRIC DISORDER OF PUBLIC HEALTH IMPORTANCE AFFECTING >1% OF THE SWEDISH POPULATION. DESPITE PROGRESS, PATIENTS STILL SUFFER FROM CHRONIC MOOD SWITCHES WITH POTENTIAL SEVERE CONSEQUENCES. THUS, EARLY DETECTION, DIAGNOSIS AND INITIATION OF CORRECT TREATMENT ARE CRITICAL. CULTURED ADIPOCYTES FROM 35 PATIENTS WITH BD AND 38 HEALTHY CONTROLS WERE ANALYSED USING SIGNAL PATHWAY REPORTER ASSAYS, THAT IS, PROTEIN KINASE C (PKC), PROTEIN KINASE A (PKA), MITOGEN-ACTIVATED PROTEIN KINASES (EXTRACELLULAR SIGNAL-REGULATED KINASE (ERK) AND C-JUN N-TERMINAL KINASE (JNK)), MYC, WNT AND P53. THE LEVELS OF ACTIVATED TARGET TRANSCRIPTIONAL FACTORS WERE MEASURED IN ADIPOCYTES BEFORE AND AFTER STIMULATION WITH LITHIUM AND ESCITALOPRAM. VARIATIONS WERE ANALYSED IN THE LOCI OF 25 DIFFERENT SINGLE-NUCLEOTIDE POLYMORPHISMS (SNPS). ACTIVATION OF INTRACELLULAR SIGNALS IN SEVERAL PATHWAYS ANALYSED WERE SIGNIFICANTLY HIGHER IN PATIENTS THAN IN HEALTHY CONTROLS UPON DRUG STIMULATION, ESPECIALLY WITH ESCITALOPRAM STIMULATION OF PKC, JNK AND MYC, AS WELL AS LITHIUM-STIMULATED PKC, WHEREAS NO MEANINGFUL DIFFERENCE WAS OBSERVED BEFORE STIMULATION. UNIVARIATE ANALYSES OF CONTINGENCY TABLES FOR 80 CATEGORICAL SNP RESULTS VERSUS DIAGNOSES SHOWED A SIGNIFICANT LINK WITH THE ANK3 GENE (RS10761482; LIKELIHOOD RATIO CHI(2)=4.63; P=0.031). IN A MULTIVARIATE ORDINAL LOGISTIC FIT FOR DIAGNOSIS, A BACKWARD STEPWISE PROCEDURE SELECTED ANK3 AS THE REMAINING SIGNIFICANT PREDICTOR. COMPARISON OF THE ESCITALOPRAM-STIMULATED PKC ACTIVITY AND THE ANK3 GENOTYPE SHOWED THEM TO ADD THEIR SHARE OF THE DIAGNOSTIC VARIANCE, WITH NO INTERACTION (15% OF VARIANCE EXPLAINED, P<0.002). THE STUDY IS CROSS-SECTIONAL WITH NO LONGITUDINAL FOLLOW-UP. COHORTS ARE RELATIVELY SMALL WITH NO MEDICATION-FREE PATIENTS, AND THERE ARE NO 'ILL PATIENT' CONTROLS. IT TAKES 3 TO 4 WEEKS OF CULTURE TO EXPAND ADIPOCYTES THAT MAY CHANGE EPIGENETIC PROFILES BUT REMOVE THE POSSIBILITY OF MEDICATION EFFECTS. ABNORMALITIES IN THE REACTIVITY OF INTRACELLULAR SIGNAL PATHWAYS TO STIMULATION AND THE ANK3 GENOTYPE MAY BE ASSOCIATED WITH PATHOGENESIS OF BD. ALGORITHMS USING BIOLOGICAL PATTERNS SUCH AS PATHWAY REACTIVITY TOGETHER WITH STRUCTURAL GENETIC SNP DATA MAY PROVIDE OPPORTUNITIES FOR EARLIER DETECTION AND EFFECTIVE TREATMENT OF BD.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
18  6368  26 THE ROLE OF MICRORNAS IN CHRONIC PSEUDOMONAS LUNG INFECTION IN CYSTIC FIBROSIS. BACKGROUND: CYSTIC FIBROSIS (CF) IS THE MOST COMMON LIFE LIMITING GENETIC DISORDER, CHARACTERIZED BY CHRONIC RESPIRATORY FAILURE SECONDARY TO INFLAMMATION AND CHRONIC BACTERIAL LUNG INFECTION. PSEUDOMONAS AERUGINOSA LUNG INFECTION IS ASSOCIATED WITH MORE SEVERE LUNG DISEASE AND RAPID PROGRESSION OF RESPIRATORY FAILURE WHEN COMPARED TO STAPHYLOCOCCUS AUREUS INFECTION. WE HYPOTHESIZED THAT A SPECIFIC SIGNATURE OF EPIGENETIC FACTORS TARGETING SPECIFIC GENE TRANSCRIPTS CONTRIBUTES TO THE INCREASED MORBIDITY SEEN IN CF PATIENTS WITH CHRONIC PSEUDOMONAS INFECTION. METHODS: WE COLLECTED EXHALED BREATH CONDENSATE (EBC) FROM 27 SUBJECTS AND EVALUATED MIRNA SIGNATURES IN THESE SAMPLES USING COMMERCIAL PCR ARRAY. WE IDENTIFIED PREDICTED MRNA TARGETS AND ASSOCIATED SIGNALING PATHWAYS USING INGENUITY PATHWAY ANALYSIS. RESULTS: WE FOUND 11 DIFFERENTIALLY EXPRESSED MIRNAS IN EBC OF PATIENTS INFECTED WITH PSEUDOMONAS AERUGINOSA COMPARED TO EBC FROM CF PATIENTS WHO WERE NOT CHRONICALLY INFECTED WITH PSEUDOMONAS AERUGINOSA (P < 0.05). SIX OF THESE MIRNAS (HSA-MIRNA-1247, HSA-MIRNA-1276, HSA-MIRNA-449C, HSA-MIRNA-3170, HSA-MIRNA-432-5P AND HSA-MIR-548) WERE SIGNIFICANTLY DIFFERENT IN THE CF PSEUDOMONAS POSITIVE GROUP WHEN COMPARED TO BOTH THE CF PSEUDOMONAS NEGATIVE GROUP AND HEALTHY CONTROL GROUP. INGENUITY PATHWAY ANALYSIS (IPA) REVEALED ORGANISMAL INJURY AND ABNORMALITIES, REPRODUCTIVE SYSTEM DISEASE AND CANCER AS THE TOP DISEASES AND BIO FUNCTIONS ASSOCIATED WITH THESE MIRNAS. IPA ALSO DETECTED RELA, JUN, TNF, IL-10, CTNNB1, IL-13, SERPINB8, CALM1, STARD3NL, SFI1, CD55, RPS6KA4, TTC36 AND HIST1H3D AS THE TOP TARGET GENES FOR THESE MIRNAS. CONCLUSION: OUR STUDY IDENTIFIED 6 MIRNAS AS EPIGENETIC FACTORS SPECIFICALLY ASSOCIATED WITH CHRONIC PSEUDOMONAS INFECTION IN PATIENTS WITH CF.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
19  5010  30 PEROXIDATION OF LINOLEIC, ARACHIDONIC AND OLEIC ACID IN RELATION TO THE INDUCTION OF OXIDATIVE DNA DAMAGE AND CYTOGENETIC EFFECTS. IN THE PRESENT STUDY, THE POSSIBLE ROLE OF THE POLYUNSATURATED FATTY ACIDS LINOLEIC AND ARACHIDONIC ACID IN THE CHEMICAL INDUCTION OF CARCINOGENESIS HAS BEEN INVESTIGATED. ANALYSIS OF 7,8-DIHYDRO-8-OXO-2'-DEOXYGUANOSINE (8-OXODG) LEVELS IN 2'-DEOXYGUANOSINE (DG) AND ISOLATED DNA HAS DEMONSTRATED THAT LINOLEIC AND ARACHIDONIC ACID ARE CAPABLE OF INDUCING THIS SPECIFIC GENOTOXIC DAMAGE. THIS EFFECT APPEARS TO BE RELATED TO THE DEGREE OF FATTY ACID UNSATURATION, SINCE IT WAS NOT INDUCED BY MONOUNSATURATED OLEIC ACID. ENZYMATIC PEROXIDATION OF LINOLEIC AND ARACHIDONIC ACID RESULTED IN A SIGNIFICANT INCREASE IN OXIDATIVE DNA DAMAGE. STUDIES ON THE INTERFERENCE OF RADICAL SCAVENGERS WITH THE INDUCTION OF 8-OXODG IN COMBINATION WITH ELECTRON SPIN RESONANCE SPECTROSCOPY DEMONSTRATED THAT THE SUPEROXIDE ANION WAS GENERATED DURING PEROXIDATION OF THESE FATTY ACIDS AND THAT SINGLET OXYGEN IS MOST LIKELY INVOLVED IN THE FORMATION OF OXIDATIVE DNA DAMAGE. THE LEVEL OF OXIDATIVE DAMAGE IN DG AND SINGLE-STRANDED DNA WAS HIGHER AS COMPARED TO THAT IN NATIVE DNA AFTER EQUIMOLAR TREATMENT. EXPOSURE OF HUMAN LYMPHOCYTES TO LINOLEIC OR ARACHIDONIC ACID DID NOT RESULT IN A SIGNIFICANT INCREASE IN LEVELS OF 8-OXODG. THIS MAY INDICATE THAT THE RATE OF INTRACELLULAR PEROXIDATION IS RELATIVELY LOW AND/OR THAT NUCLEAR DNA IN INTACT CELLS IS EFFECTIVELY PROTECTED AGAINST GENETIC DAMAGE INDUCED BY REACTIVE OXYGEN SPECIES. IT IS THEREFORE CONCLUDED THAT RELATIVELY SHORT PERIODS OF LINOLEIC OR ARACHIDONIC ACID ADMINISTRATION ARE NOT LIKELY TO IMPOSE A DIRECT GENOTOXIC RISK. IT CAN, HOWEVER, NOT BE EXCLUDED THAT CHRONIC EXPOSURE TO POLYUNSATURATED FATTY ACIDS INDUCES OXIDATIVE DNA DAMAGE OR IS RELATED TO CANCER RISK BY EPIGENETIC MECHANISMS, AS IS ALSO INDICATED BY THE OBSERVED CYTOTOXIC EFFECTS OF LINOLEIC AND ARACHIDONIC ACID.	1994	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
20  4303  31 MICRORNA-223 INHIBITS TISSUE FACTOR EXPRESSION IN VASCULAR ENDOTHELIAL CELLS. OBJECTIVE: ATHEROSCLEROSIS IS A CHRONIC INFLAMMATORY PROCESS, IN WHICH VASCULAR ENDOTHELIAL CELLS (ECS) BECOME DYSFUNCTIONAL OWING TO THE EFFECTS OF CHEMICAL SUBSTANCES, SUCH AS INFLAMMATORY FACTOR AND GROWTH FACTORS. TISSUE FACTOR (TF) EXPRESSION IS INDUCED BY THE ABOVE CHEMICAL SUBSTANCES IN ACTIVATED ECS. TF INITIATES THROMBOSIS ON DISRUPTED ATHEROSCLEROTIC PLAQUES WHICH PLAYS AN ESSENTIAL ROLE DURING THE ONSET OF ACUTE CORONARY SYNDROMES (ACS). INCREASING EVIDENCES SUGGEST THE IMPORTANT ROLE OF MICRORNAS AS EPIGENETIC REGULATORS OF ATHEROSCLEROTIC DISEASE. THE AIM OF OUR STUDY IS TO IDENTIFY IF MICRORNA-223 (MIR-223) TARGETS TF IN ECS. METHODS AND RESULTS: BIOINFORMATIC ANALYSIS SHOWED THAT TF IS A TARGET CANDIDATE OF MIR-223. WESTERN BLOTTING ANALYSIS REVEALED THAT TUMOR NECROSIS FACTOR ALPHA (TNF-ALPHA) INCREASED TF EXPRESSION IN AORTA OF C57BL/6J MICE AND CULTURED ECS (EA.HY926 CELLS AND HUVEC) AFTER 4 H TREATMENT. IN TNF-ALPHA TREATED ECS, TF MRNA WAS ALSO INCREASED MEASURED BY REAL-TIME PCR. REAL-TIME PCR RESULTS SHOWED THAT MIR-223 LEVELS WERE DOWNREGULATED IN TNF-ALPHA-TREATED AORTA OF C57BL/6J MICE AND CULTURED ECS. TRANSFECTION OF ECS WITH MIR-223 MIMIC OR MIR-223 INHIBITOR MODIFIED TF EXPRESSION BOTH IN MRNA AND PROTEIN LEVELS. LUCIFERASE ASSAYS CONFIRMED THAT MIR-223 SUPPRESSED TF EXPRESSION BY BINDING TO THE SEQUENCE OF TF 3'-UNTRANSLATED REGIONS (3'UTR). TF PROCOAGULANT ACTIVITY WAS INHIBITED BY OVEREXPRESSING MIR-223 WITH OR WITHOUT TNF-ALPHA STIMULATION. CONCLUSIONS: MIR-223-MEDIATED SUPPRESSION OF TF EXPRESSION PROVIDES A NOVEL MOLECULAR MECHANISM FOR THE REGULATION OF COAGULATION CASCADE, AND SUGGESTS A CLUE AGAINST THROMBOGENESIS DURING THE PROCESS OF ATHEROSCLEROTIC PLAQUE RUPTURE.	2014