1  1472 127 DISTINCT MECHANISMS REGULATE IL1B GENE TRANSCRIPTION IN LYMPHOID CD4 T CELLS AND MONOCYTES. INTERLEUKIN 1BETA IS A PRO-INFLAMMATORY CYTOKINE IMPORTANT FOR BOTH NORMAL IMMUNE RESPONSES AND CHRONIC INFLAMMATORY DISEASES. THE REGULATION OF THE 31 KDA PROIL-1BETA PRECURSOR CODED BY THE IL1B GENE HAS BEEN EXTENSIVELY STUDIED IN MYELOID CELLS, BUT NOT IN LYMPHOID-DERIVED CD4 T CELLS. SURPRISINGLY, WE FOUND THAT SOME CD4 T CELL SUBSETS EXPRESS HIGHER LEVELS OF PROIL-1BETA THAN UNSTIMULATED MONOCYTES, DESPITE RELATIVELY LOW IL1B MRNA LEVELS. WE OBSERVED A SIGNIFICANT INCREASE IN IL1B TRANSCRIPTION AND TRANSLATION IN CD4 T CELLS UPON EX VIVO CD3/CD28 ACTIVATION, AND A SIMILAR ELEVATION IN THE CCR5+ EFFECTOR MEMORY POPULATION COMPARED TO CCR5- T CELLS IN VIVO. THE RAPID AND VIGOROUS INCREASE IN IL1B GENE TRANSCRIPTION FOR STIMULATED MONOCYTES HAS PREVIOUSLY BEEN ASSOCIATED WITH THE PRESENCE OF SPI-1/PU.1 (SPI1), A MYELOID-LINEAGE TRANSCRIPTION FACTOR, PRE-BOUND TO THE PROMOTER. IN THE CASE OF CD4 T CELLS, THIS INCREASE OCCURRED DESPITE THE LACK OF DETECTABLE SPI1 AT THE IL1B PROMOTER. ADDITIONALLY, WE FOUND ALTERED EPIGENETIC REGULATION OF THE IL1B LOCUS IN CD3/CD28-ACTIVATED CD4 T CELLS. UNLIKE MONOCYTES, ACTIVATED CD4 T CELLS POSSESS BIVALENT H3K4ME3+/H3K27ME3+ NUCLEOSOME MARKS AT THE IL1B PROMOTER, REFLECTING LOW TRANSCRIPTIONAL ACTIVITY. THESE RESULTS SUPPORT A MODEL IN WHICH THE IL1B GENE IN CD4 T CELLS IS TRANSCRIBED FROM A LOW-ACTIVITY BIVALENT PROMOTER INDEPENDENT OF SPI1. ACCUMULATED CYTOPLASMIC PROIL-1BETA MAY ULTIMATELY BE CLEAVED TO MATURE 17 KDA BIOACTIVE IL-1BETA, REGULATING T CELL POLARIZATION AND PATHOGENIC CHRONIC INFLAMMATION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
2  3456  32 HYPOMETHYLATION OF IL1RN AND NFKB1 GENES IS LINKED TO THE DYSBALANCE IN IL1BETA/IL-1RA AXIS IN FEMALE PATIENTS WITH TYPE 2 DIABETES MELLITUS. INFLAMMATION HAS RECEIVED CONSIDERABLE ATTENTION IN THE PATHOGENESIS OF TYPE 2 DIABETES MELLITUS (T2DM). SUPPORTING THIS CONCEPT, ENHANCED EXPRESSION OF INTERLEUKIN (IL)-1BETA AND INCREASED INFILTRATION OF MACROPHAGES ARE OBSERVED IN PANCREATIC ISLETS OF PATIENTS WITH T2DM. ALTHOUGH IL-1 RECEPTOR ANTAGONIST (IL-1RA) PLAYS A MAJOR ROLE IN CONTROLLING OF IL-1BETA-MEDIATED INFLAMMATION, ITS COUNTERACTION EFFECTS AND EPIGENETIC ALTERATIONS IN T2DM ARE LESS STUDIED. THUS, WE AIMED TO ANALYZE THE DNA METHYLATION STATUS IN IL1RN, RELA (P65) AND NFKB1 (P50) GENES IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) FROM TREATED T2DM PATIENTS (N = 35) AND AGE-/SEX- MATCHED HEALTHY CONTROLS (N = 31). PRODUCTION OF IL-1BETA AND IL-1RA WAS ANALYZED IN PLASMA AND SUPERNATANTS FROM LPS-INDUCED PBMCS. IMMUNOMODULATORY EFFECTS OF IL-1BETA AND IL-1RA WERE STUDIED ON THP-1 CELLS. AVERAGE DNA METHYLATION LEVEL OF IL1RN AND NFKB1 GENE PROMOTERS WAS SIGNIFICANTLY DECREASED IN T2DM PATIENTS IN COMPARISON WITH HEALTHY CONTROLS (P< 0.05), WHICH WAS ASSOCIATED WITH THE INCREASED IL-1RA (P< 0.001) AND IL-1BETA (P = 0.039) PLASMA LEVELS IN T2DM PATIENTS. NEGATIVE ASSOCIATION BETWEEN AVERAGE METHYLATION OF IL1RN GENE AND IL-1RA PLASMA LEVELS WERE OBSERVED IN FEMALE T2DM PATIENTS. METHYLATION OF NFKB1 GENE WAS NEGATIVELY CORRELATED WITH IL-1RA LEVELS IN THE PATIENTS AND POSITIVELY WITH IL-1BETA LEVELS IN FEMALE PATIENTS. LPS-STIMULATED PBMCS FROM FEMALE PATIENTS FAILED TO RAISE IL-1BETA PRODUCTION, WHILE THE CELLS FROM HEALTHY FEMALES INCREASED IL-1BETA PRODUCTION IN COMPARISON WITH UNSTIMULATED CELLS (P< 0.001). TAKEN TOGETHER, THE FINDINGS SUGGEST THAT HYPOMETHYLATION OF IL1RN AND NFKB1 GENE PROMOTERS MAY PROMOTE THE INCREASED IL-1BETA/IL-1RA PRODUCTION AND REGULATE CHRONIC INFLAMMATION IN T2DM. FURTHER STUDIES ARE NECESSARY TO ELUCIDATE THE CAUSAL DIRECTION OF THESE ASSOCIATIONS AND POTENTIAL ROLE OF IL-1RA IN ANTI-INFLAMMATORY PROCESSES IN TREATED PATIENTS WITH T2DM.	2020	

3  6111  29 THE EPIGENETIC ARCHITECTURE AT GENE PROMOTERS DETERMINES CELL TYPE-SPECIFIC LPS TOLERANCE. SYNOVIAL FIBROBLASTS (SF) DRIVE INFLAMMATION AND JOINT DESTRUCTION IN CHRONIC ARTHRITIS. HERE WE SHOW THAT SF POSSESS A DISTINCT TYPE OF LPS TOLERANCE COMPARED TO MACROPHAGES AND OTHER TYPES OF FIBROBLASTS. IN SF AND DERMAL FIBROBLASTS, GENES THAT WERE NON-TOLERIZABLE AFTER REPEATED LPS STIMULATION INCLUDED PRO-INFLAMMATORY CYTOKINES, CHEMOKINES AND MATRIX METALLOPROTEINASES, WHEREAS ANTI-VIRAL GENES WERE TOLERIZABLE. IN MACROPHAGES, ALL MEASURED GENES WERE TOLERIZABLE, WHEREAS IN GINGIVAL AND FORESKIN FIBROBLASTS THESE GENES WERE NON-TOLERIZABLE. REPEATED STIMULATION OF SF WITH LPS RESULTED IN LOSS OF ACTIVATING HISTONE MARKS ONLY IN PROMOTERS OF TOLERIZABLE GENES. THE EPIGENETIC LANDSCAPE AT PROMOTERS OF TOLERIZABLE GENES WAS SIMILAR IN UNSTIMULATED SF AND MONOCYTES, WHEREAS THE BASAL CONFIGURATION OF HISTONE MARKS PROFOUNDLY DIFFERED IN GENES THAT WERE NON-TOLERIZABLE IN SF ONLY. OUR DATA SUGGEST THAT THE EPIGENETIC CONFIGURATION AT GENE PROMOTERS REGULATES CELL-SPECIFIC LPS-INDUCED RESPONSES AND PRIMES SF TO SUSTAIN THEIR INFLAMMATORY RESPONSE IN CHRONIC ARTHRITIS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
4  3916  31 LINE1 CPG-DNA HYPOMETHYLATION IN GRANULOSA CELLS AND BLOOD LEUKOCYTES IS ASSOCIATED WITH PCOS AND RELATED TRAITS. CONTEXT: ALTERED GLOBAL DNA METHYLATION IS INDICATIVE OF EPIGENOMIC INSTABILITY CONCERNING CHRONIC DISEASES. INVESTIGATING ITS INCIDENCE AND ASSOCIATION WITH POLYCYSTIC OVARY SYNDROME (PCOS) IS ESSENTIAL TO UNDERSTAND THE ETIOPATHOGENESIS OF THIS DISORDER. OBJECTIVES: WE ASSESSED GLOBAL DNA METHYLATION DIFFERENCES IN PERIPHERAL BLOOD LEUKOCYTES (PBLS) AND CUMULUS GRANULOSA CELLS (CGCS) OF CONTROLS AND WOMEN WITH PCOS; AND THEIR ASSOCIATION WITH PCOS AND ITS TRAITS. DESIGN, SETTING, PARTICIPANTS, MAIN OUTCOME MEASURE: THIS STUDY INCLUDED A TOTAL OF 102 CONTROLS AND WOMEN WITH PCOS. FORTY-ONE WOMEN UNDERGOING CONTROLLED OVARIAN HYPERSTIMULATION (COH) AND 61 WOMEN NOT UNDERGOING COH WERE RECRUITED FROM IN VITRO FERTILIZATION (IVF) AND INFERTILITY CLINICS. DNA METHYLATION WAS MEASURED BY ELISA FOR 5'-METHYL-CYTOSINE CONTENT AND BISULFITE SEQUENCING OF 5'-UNTRANSLATED REGION (5'-UTR) OF LONG INTERSPERSED NUCLEOTIDE ELEMENT-1 (LINE1/L1). RESULTS: TOTAL 5'-METHYL-CYTOSINE AND L1 METHYLATION LEVELS IN PBLS AND CGCS WERE SIMILAR BETWEEN CONTROLS AND WOMEN WITH PCOS. METHYLATION ASSESSED AT CPG SITES OF L1 5'-UTR REVEALED A SINGLE CPG-SITE (CPG-4) TO BE CONSISTENTLY HYPOMETHYLATED IN PBLS OF BOTH PCOS GROUPS AND CGCS OF STIMULATED PCOS GROUP. IN UNSTIMULATED WOMEN, HYPOMETHYLATION AT CPG-4 WAS STRONGLY ASSOCIATED WITH PCOS SUSCEPTIBILITY, WHEREAS IN STIMULATED GROUP IT SHOWED STRONG ASSOCIATIONS WITH PCOS AND ITS HORMONAL TRAITS. FURTHERMORE, CGCS DEMONSTRATED CONSISTENT GLOBAL AND CPG-DNA HYPOMETHYLATION RELATIVE TO PBLS, IRRESPECTIVE OF NORMAL OR DISEASE STATES. CONCLUSION: OUR STUDY REVEALED STRONG ASSOCIATION OF SINGLE HYPOMETHYLATED CPG-SITE WITH PCOS. IDENTIFICATION AND CHARACTERIZATION OF MORE SUCH METHYL-CPG SIGNATURES IN REPETITIVE ELEMENTS IN LARGER STUDY POPULATIONS WOULD PROVIDE VALUABLE EPIGENETIC INSIGHTS INTO PCOS.	2017	
                                                                                                                                                  
5   169  35 ABNORMALITIES OF AMPK ACTIVATION AND GLUCOSE UPTAKE IN CULTURED SKELETAL MUSCLE CELLS FROM INDIVIDUALS WITH CHRONIC FATIGUE SYNDROME. BACKGROUND: POST EXERTIONAL MUSCLE FATIGUE IS A KEY FEATURE IN CHRONIC FATIGUE SYNDROME (CFS). ABNORMALITIES OF SKELETAL MUSCLE FUNCTION HAVE BEEN IDENTIFIED IN SOME BUT NOT ALL PATIENTS WITH CFS. TO TRY TO LIMIT POTENTIAL CONFOUNDERS THAT MIGHT CONTRIBUTE TO THIS CLINICAL HETEROGENEITY, WE DEVELOPED A NOVEL IN VITRO SYSTEM THAT ALLOWS COMPARISON OF AMP KINASE (AMPK) ACTIVATION AND METABOLIC RESPONSES TO EXERCISE IN CULTURED SKELETAL MUSCLE CELLS FROM CFS PATIENTS AND CONTROL SUBJECTS. METHODS: SKELETAL MUSCLE CELL CULTURES WERE ESTABLISHED FROM 10 SUBJECTS WITH CFS AND 7 AGE-MATCHED CONTROLS, SUBJECTED TO ELECTRICAL PULSE STIMULATION (EPS) FOR UP TO 24H AND EXAMINED FOR CHANGES ASSOCIATED WITH EXERCISE. RESULTS: IN THE BASAL STATE, CFS CULTURES SHOWED INCREASED MYOGENIN EXPRESSION BUT DECREASED IL6 SECRETION DURING DIFFERENTIATION COMPARED WITH CONTROL CULTURES. CONTROL CULTURES SUBJECTED TO 16 H EPS SHOWED A SIGNIFICANT INCREASE IN BOTH AMPK PHOSPHORYLATION AND GLUCOSE UPTAKE COMPARED WITH UNSTIMULATED CELLS. IN CONTRAST, CFS CULTURES SHOWED NO INCREASE IN AMPK PHOSPHORYLATION OR GLUCOSE UPTAKE AFTER 16 H EPS. HOWEVER, GLUCOSE UPTAKE REMAINED RESPONSIVE TO INSULIN IN THE CFS CELLS POINTING TO AN EXERCISE-RELATED DEFECT. IL6 SECRETION IN RESPONSE TO EPS WAS SIGNIFICANTLY REDUCED IN CFS COMPARED WITH CONTROL CULTURES AT ALL TIME POINTS MEASURED. CONCLUSION: EPS IS AN EFFECTIVE MODEL FOR ELICITING MUSCLE CONTRACTION AND THE METABOLIC CHANGES ASSOCIATED WITH EXERCISE IN CULTURED SKELETAL MUSCLE CELLS. WE FOUND FOUR MAIN DIFFERENCES IN CULTURED SKELETAL MUSCLE CELLS FROM SUBJECTS WITH CFS; INCREASED MYOGENIN EXPRESSION IN THE BASAL STATE, IMPAIRED ACTIVATION OF AMPK, IMPAIRED STIMULATION OF GLUCOSE UPTAKE AND DIMINISHED RELEASE OF IL6. THE RETENTION OF THESE DIFFERENCES IN CULTURED MUSCLE CELLS FROM CFS SUBJECTS POINTS TO A GENETIC/EPIGENETIC MECHANISM, AND PROVIDES A SYSTEM TO IDENTIFY NOVEL THERAPEUTIC TARGETS.	2015	
                                  
6  6747  28 WHOLE GENOME 5'-METHYLCYTOSINE LEVEL QUANTIFICATION IN CIRRHOTIC HCV-INFECTED EGYPTIAN PATIENTS WITH AND WITHOUT HEPATOCELLULAR CARCINOMA. DNA METHYLATION IS AN EPIGENETIC MECHANISM USED BY CELLS TO CONTROL GENE EXPRESSION. DNA METHYLATION IS A COMMONLY USED EPIGENETIC SIGNALING TOOL THAT CAN HOLD GENES IN THE "OFF" POSITION. CHRONIC INFECTION WITH HEPATITIS C VIRUS (HCV) IS CONSIDERED A MAJOR RISK FOR CHRONIC LIVER IMPAIRMENT. IT IS THE MOST COMMON LEADING CAUSE OF HCC. THE PRESENT WORK IS AIMED AT STUDYING WHOLE GENOME 5'-METHYLCYTOSINE LEVELS IN CIRRHOTIC HCV-INFECTED EGYPTIAN PATIENTS. IN THE PRESENT STUDY, 120 EGYPTIAN ADULTS WERE INCLUDED. THEY WERE DIVIDED INTO TWO GROUPS: GROUP CAPITAL I, UKRAINIAN (40 APPARENTLY HEALTHY CONTROL SUBJECTS) AND GROUP CAPITAL I, UKRAINIANCAPITAL I, UKRAINIAN (80 HCV-INFECTED PATIENTS). FURTHERMORE, GROUP II WAS SUBDIVIDED INTO 2 SUBGROUPS ACCORDING TO THE PRESENCE OF HCC IN HCV-INFECTED SUBJECTS. TO ALL STUDIED SUBJECTS, THE LEVEL OF 5-MC% WAS MEASURED IN PERIPHERAL BLOOD. IN THE PRESENT STUDY, THE MEDIAN OF 5'-METHYLCYTOSINE% IN THE CONTROL GROUP (GROUP I) WAS 2.5, IN THE HCV GROUP (GROUP IIA) WAS 2.45, AND IN THE HCC GROUP (GROUP II B) WAS 2.25. A STEPWISE DECREASE IN 5'-METHYLCYTOSINE% FROM THE CONTROL (GROUP I) TOWARD HCC (GROUP IIB) WAS OBSERVED, TAKING INTO CONSIDERATION THAT THE STEPWISE GLOBAL HYPOMETHYLATION WAS NOT STATISTICALLY SIGNIFICANT (P = 0.811). THERE WAS A NEGATIVE CORRELATION BETWEEN ALT AND 5'-METHYLCYTOSINE% (P = -0.029). FROM THIS STUDY, WE CAN CONCLUDE THAT GLOBAL DNA 5'-METHYLCYTOSINE% DOES NOT DIFFER IN HCV-INFECTED CIRRHOTIC PATIENTS AND HCC PATIENTS WHEN COMPARED TO NORMAL CONTROLS. CONSECUTIVELY, WE HAD CONCLUDED THAT THERE IS NO IMPACT OF 5'-METHYLCYTOSINE% ON THE DEVELOPMENT OF LIVER CIRRHOSIS OR HCC. MOREOVER, THE NEGATIVE CORRELATION BETWEEN 5'-METHYLCYTOSINE% AND SERUM ALT LEVEL DENOTES A TREND OF DECREASE IN 5'-METHYLCYTOSINE% WITH MORE LIVER DAMAGE.	2020	
                                                                                                                                                           
7  3460  31 HYPOMETHYLATION OF THE IL8 PROMOTER IN NASAL EPITHELIAL CELLS OF PATIENTS WITH CHRONIC RHINOSINUSITIS WITH NASAL POLYPS. BACKGROUND: IL-8 IS AN IMPORTANT CHEMOKINE IMPLICATED IN THE PATHOGENESIS OF CHRONIC RHINOSINUSITIS (CRS), BUT LITTLE IS KNOWN ABOUT EPIGENETIC REGULATION OF IL8 IN THE PATHOGENESIS OF CRS. OBJECTIVE: WE SOUGHT TO INVESTIGATE THE RELATIONSHIP BETWEEN THE DNA METHYLATION LEVEL IN THE IL8 PROXIMAL PROMOTER AND CRS IN HAN CHINESE SUBJECTS. METHODS: PATIENTS WITH CHRONIC RHINOSINUSITIS WITH NASAL POLYPS (CRSWNP; N = 187), PATIENTS WITH CHRONIC RHINOSINUSITIS WITHOUT NASAL POLYPS (CRSSNP; N = 89), AND CONTROL SUBJECTS (N = 57) WERE ENROLLED IN 2 INDEPENDENT COHORTS. PURIFIED HUMAN NASAL EPITHELIAL CELLS FROM EACH PARTICIPANT WERE ASSESSED FOR PERCENTAGE DNA METHYLATION OF CPG SITES IN THE IL8 PROXIMAL PROMOTER BY USING BISULFITE PYROSEQUENCING AND FOR FUNCTIONAL ASPECTS OF METHYLATION STATUS BY USING IN VITRO ASSAYS. RESULTS: DNA METHYLATION OF CPG SITES 1, 2, AND 3, RESPECTIVELY, IN THE IL8 PROXIMAL PROMOTER WAS SIGNIFICANTLY DECREASED IN HUMAN NASAL EPITHELIAL CELLS OF PATIENTS WITH CRSWNP COMPARED WITH THAT IN PATIENTS WITH CRSSNP (P < .001) AND CONTROL SUBJECTS (P < .001). PERCENTAGE OF DNA METHYLATION OF THE CPG3 SITE WAS CORRELATED NEGATIVELY WITH BOTH TISSUE EOSINOPHILIC CATIONIC PROTEIN (P < .01) AND MYELOPEROXIDASE (P < .05) LEVELS. IL-1BETA (P < .001) AND TNF-ALPHA (P < .01) SIGNIFICANTLY INCREASED IL8 EXPRESSION ACCOMPANIED BY A REDUCTION IN METHYLATION AT THE CPG3 SITE (P < .001). ELECTROPHORETIC MOBILITY SHIFT ASSAYS DEMONSTRATED THAT METHYLATION STATUS OF CPG3 CHANGED THE BINDING OF OCTAMER-BINDING TRANSCRIPTION FACTOR 1 AND NUCLEAR FACTOR KAPPAB. CONCLUSION: DECREASED DNA METHYLATION OF PARTICULARLY CPG SITES IN THE IL8 PROXIMAL PROMOTER MIGHT PLAY A ROLE IN THE PATHOGENESIS OF CRSWNP.	2019	
                                                                                                                                                                                                                                                                          
8   969  34 CHRONIC NON-BACTERIAL OSTEOMYELITIS IS ASSOCIATED WITH IMPAIRED SP1 SIGNALING, REDUCED IL10 PROMOTER PHOSPHORYLATION, AND REDUCED MYELOID IL-10 EXPRESSION. CHRONIC NON-BACTERIAL OSTEOMYELITIS (CNO) IS AN AUTO-INFLAMMATORY DISORDER THAT AFFECTS THE SKELETAL SYSTEM. INTERLEUKIN (IL-)10 IS AN IMMUNE-MODULATORY CYTOKINE THAT CONTROLS INFLAMMATION, AND LIMITS INFLAMMATORY CYTOKINE RESPONSES. DYSREGULATION OF IL-10 EXPRESSION HAS BEEN SHOWN TO RESULT IN AUTOIMMUNE AND INFECTIOUS DISEASES. WE INVESTIGATED IL-10 EXPRESSION BY MONOCYTIC CELLS FROM CNO PATIENTS AND CONTROLS. IN RESPONSE TO STIMULATION WITH LPS, IL-10 EXPRESSION FROM CNO MONOCYTES WAS REDUCED (P<0.001). THIS WAS INDEPENDENT OF IL10 PROMOTER POLYMORPHISMS. THUS, WE INVESTIGATED SP1 RECRUITMENT TO THE IL10 PROMOTER AND SAW MARKEDLY REDUCED BINDING IN CNO MONOCYTES. THIS WAS ACCOMPANIED WITH REDUCED PHOSPHORYLATION OF HISTONE H3 SERINE 10 (H3S10), AN ACTIVATING EPIGENETIC MARK. IMPAIRED RECRUITMENT OF SP1 TO THE IL10 PROMOTER, AND REDUCED H3S10 PHOSPHORYLATION, MAY BE A REFLECTION OF DEFICIENT MAPK SIGNALING IN CNO MONOCYTES IN RESPONSE TO LPS STIMULATION. THUS, WE HAVE DISCOVERED A MECHANISM THAT MAY BE CENTRAL IN THE PATHOPHYSIOLOGY OF CNO.	2011	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
9  6589  30 TUMOR NECROSIS FACTOR-ALPHA GENE PROMOTER METHYLATION IN JAPANESE ADULTS WITH CHRONIC PERIODONTITIS AND RHEUMATOID ARTHRITIS. BACKGROUND AND OBJECTIVE: OVER-EXPRESSION OF TUMOR NECROSIS FACTOR-ALPHA (TNF-ALPHA) PLAYS A PATHOLOGICAL ROLE IN CHRONIC PERIODONTITIS (CP) AND RHEUMATOID ARTHRITIS (RA), WHICH MIGHT BE REGULATED BY THE EPIGENETIC MECHANISM. THE AIM OF THE PRESENT STUDY WAS TO EVALUATE WHETHER THERE IS A UNIQUE METHYLATION PROFILE OF THE TNF-ALPHA GENE PROMOTER IN BLOOD CELLS OF INDIVIDUALS WITH CP AND RA. MATERIAL AND METHODS: THE STUDY PARTICIPANTS CONSISTED OF 30 JAPANESE ADULTS WITH RA (RA GROUP), 30 RACE-MATCHED ADULTS WITH CP ONLY (CP GROUP) AND 30 RACE-MATCHED HEALTHY CONTROLS (H GROUP). GENOMIC DNA ISOLATED FROM PERIPHERAL BLOOD WAS MODIFIED BY SODIUM BISULFITE AND ANALYZED, BY DIRECT SEQUENCING, TO INVESTIGATE DNA METHYLATION OF THE TNF-ALPHA GENE PROMOTER REGION. THE LEVEL OF TNF-ALPHA PRODUCED IN MONONUCLEAR CELLS STIMULATED WITH PORPHYROMONAS GINGIVALIS LIPOPOLYSACCHARIDE WAS DETERMINED USING ELISA. RESULTS: TWELVE CYTOSINE-GUANINE DINUCLEOTIDE (CPG) MOTIFS WERE IDENTIFIED IN THE TNF-ALPHA PROMOTER FRAGMENT FROM -343 TO +57 BP. THE CP GROUP SHOWED A SIGNIFICANTLY HIGHER METHYLATION RATE AND FREQUENCY AT -72 BP THAN THE H GROUP (P < 0.01). THE RA GROUP EXHIBITED SIGNIFICANTLY HIGHER METHYLATION RATES AT SEVEN CPG MOTIFS (-302, -163, -119, -72, -49, -38 AND +10 BP), AND SIGNIFICANTLY HIGHER METHYLATION FREQUENCIES AT SIX CPG MOTIFS (-163, -119, -72, -49, -38 AND +10 BP), THAN THE H GROUP (P < 0.01 FOR ALL COMPARISONS). THE LEVELS OF TNF-ALPHA PRODUCED WERE SIGNIFICANTLY DIFFERENT BETWEEN INDIVIDUALS WITH AND WITHOUT METHYLATION AT -163 BP (P = 0.03). CONCLUSION: THESE RESULTS SUGGEST THAT THE HYPERMETHYLATED STATUS OF CPG MOTIFS IN THE TNF-ALPHA GENE PROMOTER IN BLOOD CELLS MAY BE UNIQUE TO JAPANESE ADULTS WITH CP AND RA.	2016	
                                                                                                                                                                                                                                                
10  3199  36 HDAC1 INTERACTS WITH THE P50 NF-?B SUBUNIT VIA ITS NUCLEAR LOCALIZATION SEQUENCE TO CONSTRAIN INFLAMMATORY GENE EXPRESSION. THE NF-?B P50 SUBUNIT IS AN IMPORTANT REGULATOR OF INFLAMMATION, WITH RECENT EXPERIMENTAL EVIDENCE TO SUPPORT IT ALSO HAVING A TUMOR SUPPRESSOR ROLE. CLASSICALLY, P50 FUNCTIONS IN HETERODIMERIC FORM WITH THE RELA (P65) NF-?B SUBUNIT TO ACTIVATE INFLAMMATORY GENES. HOWEVER, P50 ALSO FORMS HOMODIMERS WHICH ACTIVELY REPRESS NF-?B-DEPENDENT INFLAMMATORY GENE EXPRESSION AND EXERT AN IMPORTANT BRAKE ON THE INFLAMMATORY PROCESS. THIS REPRESSIVE ACTIVITY OF P50:P50 IS THOUGHT TO BE IN PART MEDIATED BY AN INTERACTION WITH THE EPIGENETIC REPRESSOR PROTEIN HISTONE DEACETYLASE 1 (HDAC1). HOWEVER, NEITHER THE INTERACTION OF P50 WITH HDAC1 NOR THE REQUIREMENT OF HDAC1 FOR THE REPRESSIVE ACTIVITIES OF P50 HAS BEEN WELL DEFINED. HERE WE EMPLOYED IN SILICO PREDICTION WITH IN VITRO ASSAYS TO MAP SITES OF INTERACTION OF HDAC1 ON THE P50 PROTEIN. DIRECTED MUTAGENESIS OF ONE SUCH REGION RESULTED IN ALMOST COMPLETE LOSS OF HDAC1 BINDING TO P50. TRANSFECTED MUTANT P50 PROTEIN LACKING THE PUTATIVE HDAC1 DOCKING MOTIF RESULTED IN ENHANCED CYTOKINE AND CHEMOKINE EXPRESSION WHEN COMPARED WITH CELLS EXPRESSING A TRANSFECTED WILD TYPE P50. IN ADDITION, EXPRESSION OF THIS MUTANT P50 WAS ASSOCIATED WITH ENHANCED CHEMOATTRACTION OF NEUTROPHILS AND ACETYLATION OF KNOWN INFLAMMATORY GENES DEMONSTRATING THE LIKELY IMPORTANCE OF THE P50:HDAC1 INTERACTION FOR CONTROLLING INFLAMMATION. THESE NEW INSIGHTS PROVIDE AN ADVANCE ON CURRENT KNOWLEDGE OF THE MECHANISMS BY WHICH NF-?B-DEPENDENT GENE TRANSCRIPTION ARE REGULATED AND HIGHLIGHT THE POTENTIAL FOR MANIPULATION OF P50:HDAC1 INTERACTIONS TO BRING ABOUT EXPERIMENTAL MODULATION OF CHRONIC INFLAMMATION AND PATHOLOGIES ASSOCIATED WITH DYSREGULATED NEUTROPHIL ACCUMULATION AND ACTIVATION.	2018	
                                                                                                                                                                                                                                                                      
11  5417  37 REGULATION OF DNA METHYLATION IN RHEUMATOID ARTHRITIS SYNOVIOCYTES. RHEUMATOID ARTHRITIS (RA) IS A CHRONIC INFLAMMATORY DISEASE IN WHICH FIBROBLAST-LIKE SYNOVIOCYTES (FLS) EXHIBIT AN AGGRESSIVE PHENOTYPE. ALTHOUGH THE MECHANISMS RESPONSIBLE ARE NOT WELL DEFINED, EPIGENETIC DETERMINANTS SUCH AS DNA METHYLATION MIGHT CONTRIBUTE. DNA METHYLTRANSFERASES (DNMTS) ARE CRITICAL ENZYMES THAT ESTABLISH AND MAINTAIN DNA METHYLATION. WE EVALUATED WHETHER PROINFLAMMATORY CYTOKINES MIGHT CONTRIBUTE TO DIFFERENTIAL DNA METHYLATION PREVIOUSLY DESCRIBED IN RA FLS THROUGH ALTERED DNMT EXPRESSION. FLS WERE OBTAINED FROM RA AND OSTEOARTHRITIS (OA) SYNOVIUM AT THE TIME OF TOTAL JOINT REPLACEMENT. GENE EXPRESSION WAS DETERMINED BY QUANTITATIVE REAL-TIME PCR AND PROTEIN EXPRESSION BY WESTERN BLOT ANALYSIS. DNMT ACTIVITY WAS MEASURED WITH A FUNCTIONAL ASSAY, AND GLOBAL METHYLATION WAS DETERMINED BY AN IMMUNOASSAY THAT DETECTS METHYLCYTOSINE. RESTING EXPRESSION OF DNMT1, -3A, AND -3B MRNA WERE SIMILAR IN RA AND OA FLS. WESTERN BLOT SHOWED ABUNDANT DNMT1 AND DNMT3A PROTEIN. EXPOSURE TO IL-1 DECREASED DNMT1 AND DNMT3A MRNA EXPRESSION IN FLS. DOSE RESPONSES DEMONSTRATED DECREASED DNMT EXPRESSION AT CONCENTRATIONS AS LOW AS 1 PG/ML OF IL-1. DNMT MRNA LEVELS DECREASED RAPIDLY, WITH SIGNIFICANT SUPPRESSION AFTER 2-8 H OF IL-1 STIMULATION. IL-1 STIMULATION OF OA FLS DID NOT AFFECT METHYLATION OF LINE1 SITES BUT LED TO DEMETHYLATION OF A CHI3L1 LOCUS THAT IS HYPOMETHYLATED IN RA FLS. CHRONIC IL-1 STIMULATION ALSO MIMICKED THE EFFECT OF A DNMT INHIBITOR ON FLS GENE EXPRESSION. EXPOSURE TO PROINFLAMMATORY MEDIATORS REVERSIBLY ALTERS DNA METHYLATION IN FLS BY DECREASING DNMT EXPRESSION AND FUNCTION. THESE DATA SUGGEST THAT IL-1 CAN POTENTIALLY IMPRINT CELLS IN CHRONIC INFLAMMATORY DISEASES.	2013	
                                                                                                                                                                                                                                                                                                                                     
12  3792  30 INTERLEUKIN-1BETA INCREASES THE RISK OF GASTRIC CANCER THROUGH INDUCTION OF ABERRANT DNA METHYLATION IN A MOUSE MODEL. INTERLEUKIN-1BETA (IL-1BETA) HAS A SIGNIFICANT ROLE IN CHRONIC GASTRIC INFLAMMATION AND MANIFESTATIONS OF GASTRIC DISEASES. THE PRESENT STUDY AIMED TO ELUCIDATE THE SPECIFIC ROLE OF IL-1BETA IN INDUCTION OF DNA METHYLATION USING IL-1 RECEPTOR TYPE 1 KNOCKOUT (IL-1R1(-)/(-)) MICE. IN THE PRESENT STUDY, WILD-TYPE (WT) AND IL-1R1(-)/(-) MICE WERE INJECTED WITH IL-1BETA (5 MICROG/KG/DAY). SERUM LEVELS OF IL-1BETA, INTERLEUKIN-6 (IL-6) AND NITRIC OXIDE (NO) WERE MEASURED BY ENZYME-LINKED IMMUNOSORBENT OR NO ASSAYS. E-CADHERIN (E-CAD) METHYLATION STATUS AND MESSENGER (M)RNA EXPRESSION OF IL-1BETA, IL-6, E-CAD AND INDUCIBLE NITRIC OXIDE SYNTHASE (INOS) WERE ANALYZED. RESULTS FROM THE PRESENT STUDY INDICATED SIGNIFICANTLY HIGHER IL-1BETA MRNA EXPRESSION (P<0.001) IN WT MICE COMPARED WITH IL-1R1(-)/(-) MICE. IL-1BETA AND IL-6 RELEASE WAS SIGNIFICANTLY INCREASED IN TREATED WT MICE COMPARED WITH IL-1R1(-)/(-) MICE AT 1 H, 4 H AND 8 H (ALL P<0.005). IL-1BETA RELEASE WAS ONLY DETECTED IN WT MICE FOLLOWING A SECOND DOSE MEASURED AT DAY 3, WEEK 1 AND WEEK 2 WHEN COMPARED WITH IL-1R1(-)/(-) MICE. PROMOTER METHYLATION OF E-CAD AND A DECREASE IN GENE EXPRESSION WAS OBSERVED IN TREATED WT MICE. MRNA EXPRESSION OF INOS IN WT MICE WAS SIGNIFICANTLY INCREASED AT WEEK 1 COMPARED WITH IL-1R1(-)/(-) MICE (P=0.0411). FURTHERMORE, A SIGNIFICANTLY INCREASED LEVEL OF NO PRODUCTION WAS OBSERVED IN TREATED WT MICE (P<0.005 AT 8 H AND WEEK 1; P<0.001 AT 4 H AND DAY 3) WHEN COMPARED WITH IL-1R1(-)/(-) MICE. THE PRESENT RESULTS INDICATED THAT IL-1BETA WAS ABLE TO DIRECTLY INDUCE DNA METHYLATION, WHICH MAY LINK INFLAMMATION-INDUCED EPIGENETIC CHANGES AND THE DEVELOPMENT OF GASTRIC DISEASES.	2016	
                                                                                                                                                                                                                                                                                                                   
13  4242  25 METHYLATION STATUS OF ALU AND LINE-1 INTERSPERSED REPETITIVE SEQUENCES IN BEHCET'S DISEASE PATIENTS. BEHCET'S DISEASE (BD) IS A MULTISYSTEM CHRONIC INFLAMMATORY DISEASE. THE PATHOLOGY IS BELIEVED TO INVOLVE BOTH GENETIC SUSCEPTIBILITY AND ENVIRONMENTAL FACTORS. HYPOMETHYLATION LEADING TO ACTIVATION OF INTERSPERSED REPETITIVE SEQUENCES (IRSS) SUCH AS LINE-1 AND ALU CONTRIBUTES TO THE PATHOLOGIES OF AUTOIMMUNE DISEASES AND CANCER. HEREIN, THE EPIGENETIC CHANGES OF IRSS IN BD WERE EVALUATED USING COMBINED BISULFITE RESTRICTION ANALYSIS-INTERSPERSED REPETITIVE SEQUENCES (COBRA-IRS). DNA FROM NEUTROPHILS AND PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) OF BD PATIENTS WITH OCULAR INVOLVEMENT THAT WERE IN ACTIVE OR INACTIVE STATES AND HEALTHY CONTROLS WERE USED TO ANALYZE LINE-1 AND ALU METHYLATION LEVELS. FOR ALU SEQUENCES, SIGNIFICANT DIFFERENCES WERE OBSERVED IN THE FREQUENCY OF (U)C(U)C ALLELES BETWEEN PBMCS OF PATIENTS AND CONTROLS (P = 0.03), AND BETWEEN INACTIVE PATIENTS AND CONTROLS (P = 0.03). FOR NEUTROPHILS, THE FREQUENCY OF (U)C(U)C WAS SIGNIFICANTLY HIGHER BETWEEN PATIENTS AND CONTROLS (P = 0.006) AND BETWEEN INACTIVE PATIENTS AND CONTROLS (P = 0.002). THE PARTIAL METHYLATION ((U)C(M)C + (M)C(U)C) FREQUENCIES OF ALU BETWEEN INACTIVE PATIENTS AND CONTROL SAMPLES ALSO DIFFERED (P = 0.02). NO STATISTICALLY SIGNIFICANT DIFFERENCES FOR LINE-1 WERE DETECTED. THUS, CHANGES IN THE METHYLATION LEVEL OF IRS ELEMENTS MIGHT CONTRIBUTE TO THE PATHOGENESIS OF BD. THE ROLE OF ALU TRANSCRIPTS IN BD SHOULD BE INVESTIGATED FURTHER.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
14   811  25 CHANGES IN DNA METHYLATION OF TANDEM DNA REPEATS ARE DIFFERENT FROM INTERSPERSED REPEATS IN CANCER. HYPOMETHYLATION OF DNA REPETITIVE ELEMENTS IS A COMMON FINDING IN CANCER, BUT VERY LITTLE IS KNOWN ABOUT THE DNA METHYLATION CHANGES OF DIFFERENT TYPES OF DNA REPETITIVE ELEMENTS, SUCH AS INTERSPERSED REPEATS (LINE1 AND ALU YB8) AND TANDEM REPEATS (SAT-ALPHA, NBL-2 AND D4Z4). WE USED BISULFITE-PCR PYROSEQUENCING TO QUANTITATIVELY MEASURE THE DNA METHYLATION OF FIVE DIFFERENT DNA REPETITIVE ELEMENTS IN NORMAL TISSUE AND CANCER. IN ALL WE STUDIED 10 DIFFERENT TISSUES FROM FOUR INDIVIDUALS UNDERGOING AUTOPSY, 34 PAIRED NORMAL AND TUMOR TISSUES FROM PATIENTS WITH BLADDER CANCER, 58 PATIENTS WITH CHRONIC MYELOGENOUS LEUKEMIA AND 23 PATIENTS WITH ACUTE PROMYELOCYTIC LEUKEMIA. WE FOUND THAT THE DNA METHYLATION OF INTERSPERSED REPEATS (LINE1 AND ALU YB8) WAS VERY CONSISTENT FROM PERSON TO PERSON AND TISSUE TO TISSUE WHILE TANDEM DNA REPEATS APPEARED MORE VARIABLE IN NORMAL TISSUES. IN BLADDER CANCER WE FOUND CLEAR HYPOMETHYLATION OF LINE1, ALU YB8, SAT-ALPHA AND NBL-2. CONVERSELY, WE FOUND AN INCREASE IN THE DNA METHYLATION LEVELS OF D4Z4 FROM NORMAL TO CANCER. IN CONTRAST LEUKEMIA SHOWED NO SIGNIFICANT CHANGES IN THE DNA METHYLATION OF LINE1 AND ALU YB8, BUT DNA METHYLATION INCREASES IN NBL-2 AND D4Z4 TANDEM REPEATS. OUR FINDINGS SHOW THAT THE CHANGES IN DNA METHYLATION LEVELS OF INDIVIDUAL DNA REPETITIVE ELEMENTS ARE UNIQUE FOR EACH REPETITIVE ELEMENT, WHICH MAY REFLECT DISTINCT EPIGENETIC FACTORS AND MAY HAVE IMPORTANT IMPLICATIONS IN THE USE OF DNA METHYLATION OF REPETITIVE ELEMENTS AS GLOBAL DNA METHYLATION BIOMARKERS.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
15  3065  28 GENOME-WIDE DNA METHYLATION PATTERNS IN CD4+ T CELLS FROM CHINESE HAN PATIENTS WITH RHEUMATOID ARTHRITIS. INTRODUCTION: RHEUMATOID ARTHRITIS (RA) IS AN AUTOIMMUNE DISEASE THAT CAUSES CHRONIC INFLAMMATION OF THE JOINTS. RECENT EVIDENCE INDICATED THE EPIGENETIC CHANGES MAY CONTRIBUTE TO THE PATHOGENESIS OF RA. METHOD: TO UNDERSTAND THE EXTENT AND NATURE OF DYSREGULATED DNA METHYLATION IN RA CD4T CELLS, WE PERFORMED A GENOME-WIDE DNA METHYLATION STUDY IN CD4 + T CELLS IN 12 RA PATIENTS COMPARED TO 12 MATCHED NORMAL HEALTHY CONTROLS. CYTOSINE METHYLATION STATUS WAS QUANTIFIED WITH ILLUMINA METHYLATION 450K MICROARRAY. RESULT: THE DNA METHYLATION PROFILING SHOWED 383 HYPER- AND 785 HYPO-METHYLATED GENES IN THE CD4 + T CELLS OF THE RA PATIENTS (P < 3.4 X 10(-7)). GENE ONTOLOGY ANALYSIS INDICATED TRANSCRIPT ALTERNATIVE SPLICING AND PROTEIN MODIFICATION MEDIATED BY DNA METHYLATION MIGHT PLAY AN IMPORTANT ROLE IN THE PATHOGENESIS OF RA. IN ADDITION, THE RESULT SHOWED THAT HUMAN LEUKOCYTE ANTIGEN (HLA) REGION INCLUDING HLA-DRB6, HLA-DQA1 AND HLA-E WAS FREQUENTLY HYPOMETHYLATED, BUT HLA-DQB1 HYPERMETHYLATED IN CPG ISLAND REGION AND HYPOMETHYLATED IN CPG SHELF REGION IN RA PATIENTS. OUTSIDE THE MHC REGION, HDAC4, NXN, TBCD AND TMEM61 WERE THE MOST HYPERMETHYLATED GENES, WHILE ITIH3, TCN2, PRDM16, SLC1A5 AND GALNT9 ARE THE MOST HYPOMETHYLATED GENES. CONCLUSION: GENOME-WIDE DNA METHYLATION PROFILE REVEALED SIGNIFICANT DNA METHYLATION CHANGE IN CD4 + T CELLS FROM PATIENTS WITH RA.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
16  2392  33 EPIGENETIC REPRESSION OF INTERLEUKIN 2 EXPRESSION IN SENESCENT CD4+ T CELLS DURING CHRONIC HIV TYPE 1 INFECTION. THE MOLECULAR MECHANISMS FOR IL2 GENE-SPECIFIC DYSREGULATION DURING CHRONIC HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 (HIV-1) INFECTION ARE UNKNOWN. HERE, WE INVESTIGATED THE ROLE OF DNA METHYLATION IN SUPPRESSING INTERLEUKIN 2 (IL-2) EXPRESSION IN MEMORY CD4(+) T CELLS DURING CHRONIC HIV-1 INFECTION. WE OBSERVED THAT CPG SITES IN THE IL2 PROMOTER OF CD4(+) T CELLS WERE FULLY METHYLATED IN NAIVE CD4(+) T CELLS AND SIGNIFICANTLY DEMETHYLATED IN THE MEMORY POPULATIONS. INTERESTINGLY, WE FOUND THAT THE MEMORY CELLS THAT HAD A TERMINALLY DIFFERENTIATED PHENOTYPE AND EXPRESSED CD57 HAD INCREASED IL2 PROMOTER METHYLATION RELATIVE TO LESS DIFFERENTIATED MEMORY CELLS IN HEALTHY INDIVIDUALS. IMPORTANTLY, EARLY EFFECTOR MEMORY SUBSETS FROM HIV-1-INFECTED SUBJECTS EXPRESSED HIGH LEVELS OF CD57 AND WERE HIGHLY METHYLATED AT THE IL2 LOCUS. FURTHERMORE, THE INCREASED CD57 EXPRESSION ON MEMORY CD4(+) T CELLS WAS INVERSELY CORRELATED WITH IL-2 PRODUCTION. THESE DATA SUGGEST THAT DNA METHYLATION AT THE IL2 LOCUS IN CD4(+) T CELLS IS COUPLED TO IMMUNOSENESCENCE AND PLAYS A CRITICAL ROLE IN THE BROAD DYSFUNCTION THAT OCCURS IN POLYCLONAL T CELLS DURING HIV-1 INFECTION.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
17  6085  26 THE EFFECTS OF ACARBOSE ON CHEMOKINE AND CYTOKINE PRODUCTION IN HUMAN MONOCYTIC THP-1 CELLS. BACKGROUND AND OBJECTIVES: CHRONIC INFLAMMATION INDUCED BY PROINFLAMMATORY CYTOKINES AND CHEMOKINES IS POSTULATED TO BE INVOLVED IN INSULIN RESISTANCE AND BETA-CELL DYSFUNCTION IN TYPE 2 DIABETES MELLITUS (T2DM). ACARBOSE, THE ALPHA-GLUCOSIDASE INHIBITOR, IS AN ORAL ANTIDIABETIC DRUG FOR T2DM. ACARBOSE SUPPRESSES INFLAMMATORY CYTOKINE PRODUCTION IN PATIENTS WITH T2DM, THOUGH THE UNDERLYING MECHANISMS ARE UNCLEAR. IN THE PRESENT STUDY, WE AIMED TO INVESTIGATE THE ANTI-INFLAMMATORY EFFECTS AND THE EXACT MECHANISMS OF ACARBOSE IN HUMAN MONOCYTIC THP-1 CELLS. METHODS: THP-1 CELLS WERE PRETREATED WITH ACARBOSE AND THEN STIMULATED WITH LIPOPOLYSACCHARIDE (LPS). THE LEVELS OF TH1-RELATED CHEMOKINES, INCLUDING INTERFERON-GAMMA-INDUCIBLE PROTEIN-10 (IP-10), MONOCYTE CHEMOATTRACTANT PROTEIN-1 (MCP-1), TH2-RELATED CHEMOKINE MACROPHAGE-DERIVED CHEMOKINE (MDC), AND PROINFLAMMATORY CYTOKINE TUMOR NECROSIS FACTOR-ALPHA (TNF-ALPHA), WERE DETERMINED BY ENZYME-LINKED IMMUNOSORBENT ASSAY. INTRACELLULAR SIGNALING PATHWAYS WERE EXPLORED BY WESTERN BLOT ANALYSIS AND USING A CHROMATIN IMMUNOPRECIPITATION ASSAY. RESULTS: ACARBOSE SUPPRESSED THE LEVELS OF IP-10, MCP-1, MDC, AND TNF-ALPHA AND DOWNREGULATED PHOSPHORYLATION OF P38, C-JUN N-TERMINAL KINASE (JNK), EXTRACELLULAR SIGNAL-REGULATED KINASE (ERK), AND NUCLEAR FACTOR-KAPPA B-P65 (NF-KAPPAB-P65) IN LPS-STIMULATED THP-1 CELLS. ACARBOSE SUPPRESSED LPS-INDUCED ACETYLATION OF HISTONES H3 (H3) AND H4 IN THE IP-10 AND MCP-1 PROMOTER REGIONS. THESE FINDINGS REVEALED THE SUPPRESSIVE EFFECTS OF ACARBOSE ON IP-10, MCP-1, MDC, AND TNF-ALPHA PRODUCTION IN THP-1 CELLS VIA, AT LEAST PARTIALLY, THE P38, JNK, ERK, AND NF-KAPPAB-P65 PATHWAYS, AS WELL AS THROUGH EPIGENETIC REGULATION VIA HISTONE H3 AND H4 ACETYLATION. CONCLUSION: OUR STUDY POINTS TO THE THERAPEUTIC ANTI-INFLAMMATORY POTENTIAL OF ACARBOSE.	2019	
                                                                                                                                                                         
18  3783  27 INTERFERON-GAMMA PROMOTER HYPOMETHYLATION AND INCREASED EXPRESSION IN CHRONIC PERIODONTITIS. AIM: THE GOAL OF THIS INVESTIGATION WAS TO DETERMINE WHETHER EPIGENETIC MODIFICATIONS IN THE IFNG PROMOTER ARE ASSOCIATED WITH AN INCREASE OF IFNG TRANSCRIPTION IN DIFFERENT STAGES OF PERIODONTAL DISEASES. MATERIALS AND METHODS: DNA WAS EXTRACTED FROM GINGIVAL BIOPSY SAMPLES COLLECTED FROM 47 TOTAL SITES FROM 47 DIFFERENT SUBJECTS: 23 PERIODONTALLY HEALTHY SITES, 12 EXPERIMENTALLY INDUCED GINGIVITIS SITES AND 12 CHRONIC PERIODONTITIS SITES. LEVELS OF DNA METHYLATION WITHIN THE IFNG PROMOTER CONTAINING SIX CPG DINUCLEOTIDES WERE DETERMINED USING PYROSEQUENCING TECHNOLOGY. INTERFERON GAMMA MRNA EXPRESSION WAS ANALYSED BY QUANTITATIVE POLYMERASE CHAIN REACTIONS USING ISOLATED RNA FROM PART OF THE BIOLOGICAL SAMPLES MENTIONED ABOVE. RESULTS: THE METHYLATION LEVEL OF ALL SIX ANALYSED CPG SITES WITHIN THE IFNG PROMOTER REGION IN THE PERIODONTITIS BIOPSIES 52% [INTERQUARTILE RANGE, IQR (43.8%, 63%)] WAS SIGNIFICANTLY LOWER THAN PERIODONTALLY HEALTHY SAMPLES 62% [IQR (51.3%, 74%)], P=0.007 AND GINGIVITIS BIOPSIES 63% [IQR (55%, 74%)], P=0.02. THE TRANSCRIPTIONAL LEVEL OF IFNG IN PERIODONTITIS BIOPSIES WAS 1.96-FOLD AND SIGNIFICANTLY HIGHER THAN TISSUES WITH PERIODONTAL HEALTH (P=0.04). ALTHOUGH THE MRNA LEVEL FROM EXPERIMENTAL GINGIVITIS SAMPLES EXHIBITED AN 8.5-FOLD INCREASE AS COMPARED WITH PERIODONTALLY HEALTHY SAMPLES, NO SIGNIFICANT METHYLATION DIFFERENCE WAS OBSERVED IN EXPERIMENTAL GINGIVITIS SAMPLE. CONCLUSIONS: A HYPOMETHYLATION PROFILE WITHIN IFNG PROMOTER REGION IS RELATED TO AN INCREASE OF IFNG TRANSCRIPTION PRESENT IN THE CHRONIC PERIODONTITIS BIOPSIES, WHILE SUCH AN INCREASE OF IFNG IN EXPERIMENTALLY INDUCED GINGIVITIS SEEMS INDEPENDENT OF PROMOTER METHYLATION ALTERATION.	2010	
                                                                                                                                                                                                                                                                                                                       
19  4831  36 OLIGOMERIC S100A4 IS ASSOCIATED WITH MONOCYTE INNATE IMMUNE MEMORY AND BYPASS OF TOLERANCE TO SUBSEQUENT STIMULATION WITH LIPOPOLYSACCHARIDES. OBJECTIVES: MOST DAMPS IN INFLAMMATORY DISEASES ARE TLR2- AND TLR4-LIGANDS AND ACCORDING TO THE CURRENT CONCEPT, REPEATED STIMULI WOULD RESULT IN TOLERANCE. AIMS OF THE STUDY WERE TO VERIFY THIS ASSUMPTION, TO INVESTIGATE WHETHER EPIGENETIC EFFECTORS ARE INVOLVED AND TO EXPLORE THE SITUATION IN RHEUMATOID ARTHRITIS (RA). METHODS: A TRAINED IMMUNITY (TI) AND TOLERANCE PROTOCOL WAS ESTABLISHED USING PERIPHERAL BLOOD MONOCYTES FROM HEALTHY DONORS, BETA-GLUCAN AND LIPOPOLYSACCHARIDE (LPS). THE TRAINING OR TOLERANCE CAPACITIES OF RA-RELEVANT DAMPS WERE TESTED. RESULTS: BETA-GLUCAN-, OS100A4-, HMBG1-, AND HSP90-PRETREATED MONOCYTES SHOWED INCREASED IL-6 RESPONSES TO LPS RE-STIMULATION. BETA-GLUCAN, OS100A AND TENASCIN C INDUCED TRAINING OF MONOCYTES TO RELEASE MORE TNFALPHA. IN COMPARISON TO BETA-GLUCAN, MOST DAMPS TESTED INDUCED LESS TI, WITH EXCEPTION OF OS100A4. MONOCYTES EXPOSED TO OS100A4 SHOWED INCREASED IL-1BETA, IL-6, AND TNFALPHA IN RESPONSE TO LPS, IN SPITE THAT BOTH STIMULATE TLR4. RNASEQ UPON BETA-GLUCAN OR OS100A4 REVEALED SIMILAR CHANGES IN CHEMOKINES/CYTOKINES AND EPIGENETIC EFFECTORS; 17 EPIGENETIC EFFECTORS CORRELATED WITH CHEMOKINE/CYTOKINE GENE EXPRESSION; PRDM8 WAS ASSOCIATED WITH MORE CHEMOKINE AND CYTOKINE TRANSCRIPTS. KNOCKDOWN OF PRDM8 ABOLISHED TI INDUCED BY OS100A4. IN RA, PLASMA S100A4 CORRELATED WITH INCREASED CSF2, AND INCREASED PRDM8 TRANSCRIPTION IN RA MONOCYTES WAS ASSOCIATED WITH INCREASED PLASMA CCL5 AND IL-6, AS WELL AS THERAPY-RESISTANCE. CONCLUSION: BYPASS OF TOLERANCE BY DAMPS MIGHT BE A PHENOMENON AS IMPORTANT AS TI, SINCE IT COULD EXPLAIN HOW CHRONIC INFLAMMATION CAN BE MAINTAINED IN SPITE OF AN ENVIRONMENT WITH MULTIPLE TLR2/TLR4-LIGANDS. IN RA MONOCYTES, A PRDM8-DEPENDENT TI MECHANISM COULD BE RESPONSIBLE FOR SUSTAINED CHEMOKINE/CYTOKINES LEVELS.	2019	
                                                                                                                                                           
20   428  27 ANTI-INFLAMMATORY ACTIVITY OF MIODESIN: MODULATION OF INFLAMMATORY MARKERS AND EPIGENETIC EVIDENCE. PURPOSE: TO INVESTIGATE THE EFFECTS OF A COMBINED HERBAL MEDICINE MIODESIN ON THE INFLAMMATORY RESPONSE OF KEY CELLS INVOLVED IN THE ACUTE AND CHRONIC INFLAMMATORY PROCESSES AS WELL AS THE POSSIBLE EPIGENETIC INVOLVEMENT. METHODS: AFTER THE ESTABLISHMENT OF THE IC(50) DOSE, THE CHONDROCYTE, KERATINOCYTE, AND MACROPHAGE CELL LINES WERE PRETREATED FOR 2 HOURS WITH MIODESIN (200 MUG/ML) AND STIMULATED WITH LPS (1 MUG/ML) FOR 24 HOURS. THE SUPERNATANT WAS USED TO MEASURE THE LEVELS OF CYTOKINES (IL-1BETA, IL-6, IL-8, AND TNF-ALPHA) AND CHEMOKINES (CCL2, CCL3, AND CCL5), AND THE CELLS WERE USED TO EXTRACT THE MRNA FOR THE TRANSCRIPTION FACTOR (NF-KAPPABETA), INFLAMMATORY ENZYMES (COX-1, COX-2, PLA2, AND INOS), AND CHEMOKINES (CCL2, CCL3, AND CCL5). RESULTS: MIODESIN INHIBITED THE RELEASE OF LPS-INDUCED CYTOKINES (IL-1BETA, IL-6, IL-8, AND TNF-ALPHA; P < 0.01) AND CHEMOKINES (CCL2, CCL3, AND CCL5; P < 0.01) AND THE EXPRESSION OF THE TRANSCRIPTION FACTOR (NF-KAPPABETA; P < 0.01), INFLAMMATORY ENZYMES (COX-1, COX-2, PLA2, INOS; P < 0.01), AND CHEMOKINES (CCL2, CCL3, AND CCL5; P < 0.01). IN ADDITION, THE EVALUATION OF EPIGENETIC MECHANISM REVEALED THAT MIODESIN DID NOT INDUCE CHANGES IN DNA METHYLATION, ASSURING THE GENETIC SAFENESS OF THE COMPOUND IN TERMS OF THE INFLAMMATORY RESPONSE. CONCLUSIONS: MIODESIN PRESENTS ANTI-INFLAMMATORY PROPERTIES, INHIBITING HYPERACTIVATION OF CHONDROCYTES, KERATINOCYTES, AND MACROPHAGES, INVOLVING EPIGENETICS IN SUCH EFFECTS.	2020