1  4989 117 PCSK9 IS INCREASED IN CEREBROSPINAL FLUID OF INDIVIDUALS WITH ALCOHOL USE DISORDER. BACKGROUND: RECENT STUDIES HAVE SHOWN THAT ALCOHOL USE AFFECTS THE REGULATION AND EXPRESSION OF PROPROTEIN CONVERTASE SUBTILISIN/KEXIN 9 (PCSK9). WHILE A MAJOR ROLE OF PCSK9 IN HEPATIC FUNCTION AND LIPID REGULATION HAS BEEN CLEARLY ESTABLISHED, OTHER PLEIOTROPIC EFFECTS REMAIN POORLY UNDERSTOOD. EXISTING RESEARCH SUGGESTS A POSITIVE ASSOCIATION BETWEEN PCSK9 EXPRESSION IN THE BRAIN AND PSYCHOPATHOLOGY, WITH INCREASED LEVELS OF PCSK9 IN THE CEREBROSPINAL FLUID (CSF) OF INDIVIDUALS WITH DEMENTIA AND EPIGENETIC MODIFICATIONS OF PCSK9 ASSOCIATED WITH ALCOHOL USE DISORDER (AUD). IN THIS STUDY, WE HYPOTHESIZED THAT CHRONIC ALCOHOL USE WOULD INCREASE PCSK9 EXPRESSION IN CSF. METHODS: PCSK9 LEVELS IN CSF WERE MEASURED IN INDIVIDUALS WITH AUD (N = 42) ADMITTED TO AN INPATIENT REHABILITATION PROGRAM AND CONTROLS (N = 25). CSF SAMPLES IN AUD WERE ASSESSED AT 2 TIME POINTS, AT DAY 5 AND DAY 21 AFTER ADMISSION. FURTHERMORE, PLASMA SAMPLES WERE COLLECTED AND MEASURED FROM THE INDIVIDUALS WITH AUD. RESULTS: PCSK9 IN CSF WAS SIGNIFICANTLY INCREASED IN THE AUD GROUP AT DAY 5 AND DAY 21 COMPARED TO THE CONTROLS (P < 0.0001). PLASMA PCSK9 LEVELS WERE CORRELATED POSITIVELY WITH CSF PCSK9 LEVELS IN AUD (P = 0.0493). CONCLUSIONS: OUR DATA SUGGEST THAT PCSK9 IS ELEVATED IN THE CSF OF INDIVIDUALS WITH AUD, WHICH MAY INDICATE A POTENTIAL ROLE OF PCSK9 IN AUD. ADDITIONAL STUDIES ARE NECESSARY TO FURTHER ELUCIDATE THE FUNCTIONS OF PCSK9 IN THE BRAIN.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
2  5285  39 PROPROTEIN CONVERTASE SUBTILISIN/KEXIN TYPE 9 (PCSK9) IN THE BRAIN AND RELEVANCE FOR NEUROPSYCHIATRIC DISORDERS. PROPROTEIN CONVERTASE SUBTILISIN/KEXIN TYPE 9 (PCSK9) HAS LONG BEEN STUDIED IN THE LIVER DUE TO ITS REGULATION OF PLASMA LOW-DENSITY LIPOPROTEIN CHOLESTEROL (LDL-C) AND ITS CAUSAL ROLE IN FAMILIAL HYPERCHOLESTEROLEMIA. ALTHOUGH PCSK9 WAS FIRST DISCOVERED IN CEREBELLAR NEURONS UNDERGOING APOPTOSIS, ITS FUNCTION IN THE CENTRAL NERVOUS SYSTEM (CNS) IS LESS CLEAR. PCSK9 HAS BEEN SHOWN TO BE INVOLVED IN NEURONAL DIFFERENTIATION, LDL RECEPTOR FAMILY METABOLISM, APOPTOSIS, AND INFLAMMATION IN THE BRAIN, BUT IN VITRO AND IN VIVO STUDIES OFFER CONTRADICTORY FINDINGS. PCSK9 EXPRESSION IN THE ADULT BRAIN IS LOW BUT IS HIGHLY UPREGULATED DURING DISEASE STATES. CEREBRAL SPINAL FLUID (CSF) PCSK9 CONCENTRATIONS ARE CORRELATED WITH NEURAL TUBE DEFECTS AND NEURODEGENERATIVE DISEASES IN HUMAN PATIENTS. EPIGENETIC STUDIES REVEAL THAT CHRONIC ALCOHOL USE MAY MODULATE METHYLATION OF THE PCSK9 GENE AND GENETIC STUDIES SHOW THAT PATIENTS WITH GAIN-OF-FUNCTION PCSK9 VARIANTS HAVE HIGHER LDL-C AND AN INCREASED RISK OF ISCHEMIC STROKE. EARLY SAFETY STUDIES OF THE PCSK9 INHIBITORS EVOLOCUMAB AND ALIROCUMAB, USED TO TREAT HYPERCHOLESTEROLEMIA, HINTED THAT PCSK9 INHIBITION MAY NEGATIVELY IMPACT COGNITION BUT MORE RECENT, LONGER-TERM CLINICAL TRIALS FOUND NO ADVERSE NEUROCOGNITIVE EVENTS. THE PURPOSE OF THIS REVIEW IS TO ELUCIDATE THE ROLE OF PCSK9 IN THE BRAIN, PARTICULARLY ITS ROLE IN DISEASE PATHOGENESIS.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
3  4640  34 NEURONAL NUCLEAR CALCIUM SIGNALING SUPPRESSION OF MICROGLIAL REACTIVITY IS MEDIATED BY OSTEOPROTEGERIN AFTER TRAUMATIC BRAIN INJURY. BACKGROUND: TRAUMATIC BRAIN INJURY (TBI) IS CHARACTERIZED BY MASSIVE CHANGES IN NEURONAL EXCITATION, FROM ACUTE EXCITOTOXICITY TO CHRONIC HYPER- OR HYPOEXCITABILITY. NUCLEAR CALCIUM SIGNALING PATHWAYS ARE INVOLVED IN TRANSLATING CHANGES IN SYNAPTIC INPUTS AND NEURONAL ACTIVITY INTO DISCRETE TRANSCRIPTIONAL PROGRAMS WHICH NOT ONLY AFFECT NEURONAL SURVIVAL AND SYNAPTIC INTEGRITY, BUT ALSO THE CROSSTALK BETWEEN NEURONS AND GLIAL CELLS. HERE, WE REPORT THE EFFECTS OF BLUNTING NEURONAL NUCLEAR CALCIUM SIGNALS IN THE CONTEXT OF TBI. METHODS: WE USED AAV VECTORS TO EXPRESS THE GENETICALLY ENCODED AND NUCLEAR-TARGETED CALCIUM BUFFER PARVALBUMIN (PV.NLS.MCHERRY) OR THE CALCIUM/CALMODULIN BUFFER CAMBP4.MCHERRY IN NEURONS ONLY. UPON TBI, THE EXTENT OF NEUROINFLAMMATION, NEURONAL DEATH AND SYNAPTIC LOSS WERE ASSESSED BY IMMUNOHISTOCHEMISTRY AND TARGETED TRANSCRIPTOME ANALYSIS. MODULATION OF THE OVERALL LEVEL OF NEURONAL ACTIVITY WAS ACHIEVED BY PSAM/PSEM CHEMOGENETICS TARGETED TO PARVALBUMIN INTERNEURONS. THE FUNCTIONAL IMPACT OF NEURONAL NUCLEAR CALCIUM BUFFERING IN TBI WAS ASSESSED BY QUANTIFICATION OF SPONTANEOUS WHISKING. RESULTS: BUFFERING NEURONAL NUCLEAR CALCIUM UNEXPECTEDLY RESULTED IN A MASSIVE AND LONG-LASTING INCREASE IN THE RECRUITMENT OF REACTIVE MICROGLIA TO THE INJURY SITE, WHICH WAS CHARACTERIZED BY A DISEASE-ASSOCIATED AND PHAGOCYTIC PHENOTYPE. THIS EFFECT WAS ACCOMPANIED BY A SUBSTANTIAL SURGE IN SYNAPTIC LOSS AND SIGNIFICANTLY REDUCED WHISKING ACTIVITY. TRANSCRIPTOME ANALYSIS REVEALED A COMPLEX EFFECT OF TBI IN THE CONTEXT OF NEURONAL NUCLEAR CALCIUM BUFFERING, WITH UPREGULATION OF COMPLEMENT FACTORS, CHEMOKINES AND INTERFERON-RESPONSE GENES, AS WELL AS THE DOWNREGULATION OF SYNAPTIC GENES AND EPIGENETIC REGULATORS COMPARED TO CONTROL CONDITIONS. NOTABLY, NUCLEAR CALCIUM BUFFERING LED TO A SUBSTANTIAL LOSS IN NEURONAL OSTEOPROTEGERIN (OPG), WHEREAS STIMULATION OF NEURONAL FIRING INDUCED OPG EXPRESSION. VIRAL RE-EXPRESSION OF OPG RESULTED IN DECREASED MICROGLIAL RECRUITMENT AND SYNAPTIC LOSS. OPG UPREGULATION WAS ALSO OBSERVED IN THE CSF OF HUMAN TBI PATIENTS, UNDERSCORING ITS TRANSLATIONAL VALUE. CONCLUSION: NEURONAL NUCLEAR CALCIUM SIGNALS REGULATE THE DEGREE OF MICROGLIAL RECRUITMENT AND REACTIVITY UPON TBI VIA, AMONG OTHERS, OSTEOPROTEGERIN SIGNALS. OUR FINDINGS SUPPORT A MODEL WHEREBY NEURONAL ACTIVITY ALTERED AFTER TBI EXERTS A POWERFUL IMPACT ON THE NEUROINFLAMMATORY CASCADE, WHICH IN TURN CONTRIBUTES TO THE OVERALL LOSS OF SYNAPSES AND FUNCTIONAL IMPAIRMENT.	2022	
                                                                                                                                                                                                                                                                                                             
4  4257  41 METHYLOMIC PROFILING AND REPLICATION IMPLICATES DEREGULATION OF PCSK9 IN ALCOHOL USE DISORDER. ALCOHOL USE DISORDER (AUD) IS A COMMON AND CHRONIC DISORDER WITH SUBSTANTIAL EFFECTS ON PERSONAL AND PUBLIC HEALTH. THE UNDERLYING PATHOPHYSIOLOGY IS POORLY UNDERSTOOD BUT STRONG EVIDENCE SUGGESTS SIGNIFICANT ROLES OF BOTH GENETIC AND EPIGENETIC COMPONENTS. GIVEN THAT ALCOHOL AFFECTS MANY ORGAN SYSTEMS, WE PERFORMED A CROSS-TISSUE AND CROSS-PHENOTYPIC ANALYSIS OF GENOME-WIDE METHYLOMIC VARIATION IN AUD USING SAMPLES FROM 3 DISCOVERY, 4 REPLICATION, AND 2 TRANSLATIONAL COHORTS. WE IDENTIFIED A DIFFERENTIALLY METHYLATED REGION IN THE PROMOTER OF THE PROPROTEIN CONVERTASE SUBTILISIN/KEXIN 9 (PCSK9) GENE THAT WAS ASSOCIATED WITH DISEASE PHENOTYPES. BIOLOGICAL VALIDATION SHOWED THAT PCSK9 PROMOTER METHYLATION IS CONSERVED ACROSS TISSUES AND POSITIVELY CORRELATED WITH EXPRESSION. REPLICATION IN AUD DATASETS CONFIRMED PCSK9 HYPOMETHYLATION AND A TRANSLATIONAL MOUSE MODEL OF AUD SHOWED THAT ALCOHOL EXPOSURE LEADS TO PCSK9 DOWNREGULATION. PCSK9 IS PRIMARILY EXPRESSED IN THE LIVER AND REGULATES LOW-DENSITY LIPOPROTEIN CHOLESTEROL (LDL-C). OUR FINDING OF ALCOHOL-INDUCED EPIGENETIC REGULATION OF PCSK9 REPRESENTS ONE OF THE UNDERLYING MECHANISMS BETWEEN THE WELL-KNOWN EFFECTS OF ALCOHOL ON LIPID METABOLISM AND CARDIOVASCULAR RISK, WITH LIGHT ALCOHOL USE GENERALLY BEING PROTECTIVE WHILE CHRONIC HEAVY USE HAS DETRIMENTAL HEALTH OUTCOMES.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
5  6302  26 THE PUTATIVE ROLE OF VIRUSES, BACTERIA, AND CHRONIC FUNGAL BIOTOXIN EXPOSURE IN THE GENESIS OF INTRACTABLE FATIGUE ACCOMPANIED BY COGNITIVE AND PHYSICAL DISABILITY. PATIENTS WHO PRESENT WITH SEVERE INTRACTABLE APPARENTLY IDIOPATHIC FATIGUE ACCOMPANIED BY PROFOUND PHYSICAL AND OR COGNITIVE DISABILITY PRESENT A SIGNIFICANT THERAPEUTIC CHALLENGE. THE EFFECT OF PSYCHOLOGICAL COUNSELING IS LIMITED, WITH SIGNIFICANT BUT VERY SLIGHT IMPROVEMENTS IN PSYCHOMETRIC MEASURES OF FATIGUE AND DISABILITY BUT NO IMPROVEMENT ON SCIENTIFIC MEASURES OF PHYSICAL IMPAIRMENT COMPARED TO CONTROLS. SIMILARLY, EXERCISE REGIMES EITHER PRODUCE SIGNIFICANT, BUT PRACTICALLY UNIMPORTANT, BENEFIT OR PROVOKE SYMPTOM EXACERBATION. MANY SUCH PATIENTS ARE AFFORDED THE EXCLUSIONARY, NON-SPECIFIC DIAGNOSIS OF CHRONIC FATIGUE SYNDROME IF RUDIMENTARY TESTING FAILS TO DISCOVER THE CAUSE OF THEIR SYMPTOMS. MORE SOPHISTICATED INVESTIGATIONS OFTEN REVEAL THE PRESENCE OF A RANGE OF PATHOGENS CAPABLE OF ESTABLISHING LIFE-LONG INFECTIONS WITH SOPHISTICATED IMMUNE EVASION STRATEGIES, INCLUDING PARVOVIRUSES, HHV6, VARIANTS OF EPSTEIN-BARR, CYTOMEGALOVIRUS, MYCOPLASMA, AND BORRELIA BURGDORFERI. OTHER PATIENTS HAVE A HISTORY OF CHRONIC FUNGAL OR OTHER BIOTOXIN EXPOSURE. HEREIN, WE EXPLAIN THE EPIGENETIC FACTORS THAT MAY RENDER SUCH INDIVIDUALS SUSCEPTIBLE TO THE CHRONIC PATHOLOGY INDUCED BY SUCH AGENTS, HOW SUCH AGENTS INDUCE PATHOLOGY, AND, INDEED, HOW SUCH PATHOLOGY CAN PERSIST AND EVEN AMPLIFY EVEN WHEN INFECTIONS HAVE CLEARED OR WHEN BIOTOXIN EXPOSURE HAS CEASED. THE PRESENCE OF ACTIVE, REACTIVATED, OR EVEN LATENT HERPES VIRUS COULD BE A POTENTIAL SOURCE OF INTRACTABLE FATIGUE ACCOMPANIED BY PROFOUND PHYSICAL AND OR COGNITIVE DISABILITY IN SOME PATIENTS, AND THE SAME MAY BE TRUE OF PERSISTENT PARVOVIRUS B12 AND MYCOPLASMA INFECTION. A HISTORY OF CHRONIC MOLD EXPOSURE IS A FEASIBLE EXPLANATION FOR SUCH SYMPTOMS, AS IS THE PRESENCE OF B. BURGDORFERI. THE COMPLEX TROPISM, LIFE CYCLES, GENETIC VARIABILITY, AND LOW TITER OF MANY OF THESE PATHOGENS MAKES THEIR DETECTION IN BLOOD A CHALLENGE. EXAMINATION OF LYMPHOID TISSUE OR CSF IN SUCH CIRCUMSTANCES MAY BE WARRANTED.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
6  4348  37 MIR-146A DYSREGULATES ENERGY METABOLISM DURING NEUROINFLAMMATION. ALZHEIMER'S DISEASE (AD) AND OTHER NEURODEGENERATIVE DISEASES ARE CHARACTERIZED BY CHRONIC NEUROINFLAMMATION AND A REDUCTION IN BRAIN ENERGY METABOLISM. AN IMPORTANT ROLE HAS EMERGED FOR SMALL, NON-CODING RNA MOLECULES KNOWN AS MICRORNAS (MIRNAS) IN THE PATHOPHYSIOLOGY OF MANY NEURODEGENERATIVE DISORDERS. AS EPIGENETIC REGULATORS, MIRNAS POSSESS THE CAPACITY TO REGULATE AND FINE TUNE PROTEIN PRODUCTION BY INHIBITING TRANSLATION. SEVERAL MIRNAS, WHICH INCLUDE MIR-146A, ARE ELEVATED IN THE BRAIN, CSF, AND PLASMA OF AD PATIENTS. MIR-146A PARTICIPATES IN PATHWAYS THAT REGULATE IMMUNE ACTIVATION AND HAS SEVERAL MRNA TARGETS WHICH ENCODE FOR PROTEINS INVOLVED IN CELLULAR ENERGY METABOLISM. AN ADDITIONAL ROLE FOR EXTRACELLULAR VESICLES (EVS) HAS ALSO EMERGED IN THE PROGRESSION AD, AS EVS CAN TRANSFER FUNCTIONALLY ACTIVE PROTEINS AND RNAS FROM DISEASED TO HEALTHY CELLS. IN THE CURRENT STUDY, WE EXPOSED VARIOUS CELL TYPES PRESENT WITHIN THE CNS TO IMMUNOMODULATORY MOLECULES AND OBSERVED SIGNIFICANT UPREGULATION OF MIR-146A EXPRESSION, BOTH WITHIN CELLS AND WITHIN THEIR SECRETED EVS. FURTHER, WE ASSESSED THE EFFECTS OF MIR-146A OVEREXPRESSION ON BIOENERGETIC FUNCTION IN PRIMARY RAT GLIAL CELLS AND FOUND SIGNIFICANT REDUCTIONS IN OXIDATIVE PHOSPHORYLATION AND GLYCOLYSIS. LASTLY, WE CORRELATED MIR-146A EXPRESSION LEVELS WITHIN VARIOUS REGIONS OF THE AD BRAIN TO DISEASE STAGING AND FOUND SIGNIFICANT, POSITIVE CORRELATIONS. THESE NOVEL RESULTS DEMONSTRATE THAT THE MODULATION OF MIR-146A IN RESPONSE TO NEUROINFLAMMATORY STIMULI MAY MEDIATE THE LOSS OF MITOCHONDRIAL INTEGRITY AND FUNCTION IN CELLS, THEREBY CONTRIBUTING TO THE PROGRESSION OF BETA-AMYLOID AND TAU PATHOLOGY IN THE AD BRAIN. MULTIPLE INFLAMMATORY STIMULI CAN UPREGULATE MIRNA-146A EXPRESSION WITHIN NEURONS, MIXED GLIAL CELLS, AND BRAIN ENDOTHELIAL CELLS, WHICH IS EITHER RETAINED WITHIN THESE CELLS OR RELEASED FROM THEM AS EXTRACELLULAR VESICLE CARGO. THE UPREGULATION OF MIR-146A DISRUPTS CELLULAR BIOENERGETICS IN MIXED GLIAL CELLS. THIS MECHANISM MAY PLAY A CRITICAL ROLE IN THE NEUROINFLAMMATORY RESPONSE OBSERVED DURING ALZHEIMER'S DISEASE.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
7  5094  37 PLASMA EXTRACELLULAR VESICLE SUBTYPES MAY BE USEFUL AS POTENTIAL BIOMARKERS OF IMMUNE ACTIVATION IN PEOPLE WITH HIV. BACKGROUND: EXTRACELLULAR VESICLES (EVS) ARE INTERCELLULAR MESSENGERS WITH EPIGENETIC POTENTIAL SINCE THEY CAN SHUTTLE MICRORNA (MIRNA). EVS AND MIRNA PLAY A ROLE IN HUMAN IMMUNODEFICIENCY VIRUS (HIV) INFECTION IMMUNOPATHOGENESIS. CHRONIC IMMUNE ACTIVATION AND SYSTEMIC INFLAMMATION DURING HIV INFECTION DESPITE EFFECTIVE ANTIRETROVIRAL THERAPY (ART) ARE ASSOCIATED WITH NON-ACQUIRED IMMUNODEFICIENCY SYNDROME (AIDS) COMORBIDITIES IN PEOPLE LIVING WITH HIV (PLWH). ANALYSIS OF PLASMA EVS AND THEIR MIRNA CONTENT MAY BE USEFUL AS IMMUNE ACTIVATION OR INFLAMMATORY BIOMARKERS IN PLWH RECEIVING ART. IN THIS STUDY, WE HYPOTHESIZED THAT THE NUMBER, SIZE, AND MIRNA OF LARGE AND SMALL EVS COULD REFLECT IMMUNE ACTIVATION ASSOCIATED WITH AN ELEVATED CD8 T-CELL COUNT OR A LOW CD4/CD8 RATIO IN PLWH. METHODS: PLASMA EVS SUBTYPE PURIFIED FROM PLWH AND UNINFECTED CONTROLS WERE SIZED USING DYNAMIC LIGHT SCATTERING AND QUANTIFIED USING FLOW CYTOMETRY AND ACETYLCHOLINE ESTERASE (ACHE) ACTIVITY. EXPRESSION OF MATURE MIRNAS MIR-92, MIR-155, MIR-223 WAS MEASURED BY QUANTITATIVE REVERSE-TRANSCRIPTASE POLYMERASE CHAIN REACTION IN EVS AND LEUCOCYTES. RESULTS: HIV INFECTION INDUCES INCREASED PRODUCTION OF SMALL EVS IN PLASMA. EV SUBTYPES WERE DIFFERENTIALLY ENRICHED IN MIR-92, MIR-155, AND MIR-223. POSITIVE CORRELATIONS BETWEEN CD8 T-CELL COUNT AND LARGE EVS ABUNDANCE AND SMALL EVS ACHE ACTIVITY WERE OBSERVED. CD4/CD8 RATIO WAS NEGATIVELY CORRELATED WITH SMALL EV ACHE ACTIVITY, AND MIRNA-155 LEVEL PER SMALL EV WAS NEGATIVELY CORRELATED WITH CD8 T-CELL COUNT. CONCLUSIONS: THESE FINDINGS SUGGEST THAT QUANTIFYING LARGE OR SMALL EVS AND PROFILING MIRNA CONTENT PER EV MIGHT PROVIDE NEW FUNCTIONAL BIOMARKERS OF IMMUNE ACTIVATION AND INFLAMMATION.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
8  1118  27 COMPARATIVE GENOTYPING AND PHENOTYPING OF ASPERGILLUS FUMIGATUS ISOLATES FROM HUMANS, DOGS AND THE ENVIRONMENT. BACKGROUND: ASPERGILLUS FUMIGATUS IS A UBIQUITOUS SAPROTROPHIC FUNGUS AND AN OPPORTUNISTIC PATHOGEN OF HUMANS AND ANIMALS. HUMANS AND ANIMALS CAN INHALE HUNDREDS OF A. FUMIGATUS SPORES DAILY. NORMALLY THIS IS HARMLESS FOR HUMANS, BUT IN CASE OF IMMUNODEFICIENCY, INVASIVE PULMONARY ASPERGILLOSIS (IPA) CAN DEVELOP WITH A HIGH MORTALITY RATE. A. FUMIGATUS ALSO CAUSES NON-INVASIVE MYCOSES LIKE SINO-NASAL ASPERGILLOSIS (SNA) IN DOGS. RESULTS: IN THIS STUDY WE COMPARED A. FUMIGATUS ISOLATES FROM HUMANS WITH SUSPECTED IPA, DOGS WITH SNA, AND A SET OF ENVIRONMENTAL ISOLATES. PHYLOGENETIC INFERENCE BASED ON CALMODULIN (CAM) AND BETA-TUBULIN (BENA) SEQUENCES DID NOT REVEAL A. FUMIGATUS SUB-GROUPS LINKED TO THE ORIGIN OF THE ISOLATES. GENOTYPING AND MICROSATELLITE ANALYSIS SHOWED THAT EACH DOG WAS INFECTED BY ONE A. FUMIGATUS GENOTYPE, WHEREAS HUMAN PATIENTS HAD MIXED INFECTIONS. AZOLE RESISTANCE WAS DETERMINED BY ANTIFUNGAL SUSCEPTIBILITY TESTING AND SEQUENCING OF THE CYP51A GENE. A TOTAL OF 12 OUT OF 29 HUMAN ISOLATES AND 1 OUT OF 27 ENVIRONMENTAL ISOLATES WERE AZOLE RESISTANT. OF THE AZOLE RESISTANT STRAINS, 11 HUMAN ISOLATES SHOWED TR(34)/L98H (N = 6) OR TR46/Y121F/T289A (N = 5). PHENOTYPICALLY, ISOLATES FROM DOGS WERE MORE VARIABLE IN GROWTH SPEED AND MORPHOLOGY WHEN COMPARED TO THOSE ISOLATED FROM HUMAN AND THE ENVIRONMENT. CONCLUSIONS: 1. A. FUMIGATUS FROM DOGS WITH SNA ARE PHENOTYPICALLY VERY DIVERSE IN CONTRAST TO THEIR ENVIRONMENTAL AND HUMAN COUNTERPARTS. 2. PHENOTYPIC VARIABILITY CAN BE INDUCED DURING THE CHRONIC INFECTION PROCESS IN THE SINUS OF THE DOGS. THE BASIS OF THIS HETEROGENEITY MIGHT BE DUE TO GENOMIC DIFFERENCES AND/OR EPIGENETIC VARIATIONS. 3. DIFFERENCES IN DOGS IS A COULD BE A RESULT OF WITHIN-HOST ADAPTION AND MIGHT BE TRIGGERED BY ENVIRONMENTAL FACTORS IN THE SINUS, HOWEVER THIS HYPOTHESIS STILL NEEDS TO BE TESTED.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
9  5682  34 SHORT-TERM CHANGES IN GLOBAL METHYLATION AND HYDROXYMETHYLATION DURING ALCOHOL DETOXIFICATION. ALCOHOL DEPENDENCE IS A COMMON PUBLIC HEALTH PROBLEM AND EPIGENETICS MAY OFFER NEW ASPECTS IN UNDERSTANDING THE BIOLOGICAL AND GENETIC UNDERPINNINGS AND IMPROVE TREATMENT OF THIS COMPLEX DISEASE. SUPPOSEDLY, METHYLATION AND HYDROXYMETHYLATION ARE ALTERED IN BRAIN TISSUES AND IN SYNAPSE-RELATED GENES DUE TO CHRONIC ALCOHOL INTAKE AND DURING WITHDRAWAL. TO ASSESS POTENTIAL EPIGENETIC CHANGES AFTER CESSATION OF CHRONIC ALCOHOL INTAKE, WE COMPARED 23 ALCOHOL-DEPENDENT INDIVIDUALS DURING INPATIENT ALCOHOL DETOXIFICATION WITH 13 CAREFULLY MATCHED CONTROLS. BLOOD SAMPLES WERE TAKEN ON THE DAY OF ADMISSION, AFTER ONE AND AFTER TWO WEEKS AT THE END OF INPATIENT TREATMENT. GENOME-WIDE GLOBAL METHYLATION AND GLOBAL DNA HYDROXYMETHYLATION WERE COMPARED ACROSS GROUPS. THERE WERE SIGNIFICANT DIFFERENCES IN GLOBAL METHYLATION ACROSS TIME FROM ADMISSION TO ONE AND TWO WEEKS OF INPATIENT WITHDRAWAL (P < 0.001). THESE FINDINGS WERE PARALLELED TO CHANGES IN GLOBAL DNA HYDROXYMETHYLATION ACROSS TIME WHEN AGE WAS EMPLOYED AS A COFACTOR (P < 0.001). SEVERAL POTENTIALLY INFLUENCING VARIABLES LIKE SEVERITY OF WITHDRAWAL, DOSE OF WITHDRAWAL MEDICATION AND ALCOHOL INTAKE BEFORE ADMISSION DID NOT YIELD SIGNIFICANT INFLUENCE ON EPIGENETIC CHANGES. THE RESULTS CONFIRM PREVIOUS FINDINGS OF SIGNIFICANT ALTERATIONS OF EPIGENETIC PATTERNS DURING ALCOHOL INTOXICATION AND PRESENT FOR THE FIRST TIME HYDROXYMETHYLATION CHANGES IN THESE INDIVIDUALS.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
10  6039  38 THE CHEMOKINE RECEPTOR CXCR2 SUPPORTS NOCICEPTIVE SENSITIZATION AFTER TRAUMATIC BRAIN INJURY. CHRONIC PAIN AFTER TRAUMATIC BRAIN INJURY (TBI) IS VERY COMMON, BUT THE MECHANISMS LINKING TBI TO PAIN AND THE PAIN-RELATED INTERACTIONS OF TBI WITH PERIPHERAL INJURIES ARE POORLY UNDERSTOOD. CHEMOKINE RECEPTORS PLAY AN IMPORTANT ROLE IN BOTH PAIN AND BRAIN INJURY. IN THE CURRENT WORK, WE PURSUED THE HYPOTHESIS THAT THE EPIGENETICALLY REGULATED CXC CHEMOKINE RECEPTOR 2 (CXCR2) IS A CRUCIAL MODULATOR OF NOCICEPTIVE SENSITIZATION INDUCED BY TBI. FOR THESE STUDIES, WE USED THE RAT LATERAL FLUID PERCUSSION MODEL OF TBI. HISTONE ACTYLTRANSFERASE ACTIVITY WAS BLOCKED USING ANACARDIC ACID BEGINNING IMMEDIATELY FOLLOWING INJURY, OR DELAYED FOR SEVEN DAYS PRIOR TO ADMINISTRATION. THE SELECTIVE CXCR2 ANTAGONIST SCH527123 ADMINISTERED SYSTEMICALLY OR INTRATHECALLY WAS USED TO PROBE THE ROLE OF CHEMOKINE SIGNALING ON MECHANICAL HINDPAW SENSITIZATION AFTER TBI. THE EXPRESSION OF THE CXCR2 RECEPTOR WAS ACCOMPLISHED USING REAL-TIME PCR, IMMUNOHISTOCHEMISTRY, AND WESTERN BLOTTING, WHILE EPIGENETIC REGULATION WAS ASSESSED USING CHROMATIN IMMUNOPRECIPITATION ASSAY. THE SPINAL LEVELS OF SEVERAL PAIN-RELATED MEDIATORS INCLUDING CXCL1, AN ENDOGENOUS LIGAND FOR CXCR2, AS WELL AS BRAIN-DERIVED NEUROTROPHIC FACTOR AND PRODYNORPHIN WERE MEASURED BY ENZYME-LINKED IMMUNOSORBENT ASSAY. WE OBSERVED THAT ANACARDIC ACID POTENTLY BLOCKED AND REVERSED MECHANICAL HINDPAW SENSITIZATION AFTER TBI. THE SAME DRUG WAS ABLE TO PREVENT THE UPREGULATION OF CXCR2 AFTER TBI, BUT DID NOT AFFECT THE SPINAL EXPRESSION OF OTHER PAIN MEDIATORS. ON THE OTHER HAND, BOTH SYSTEMICALLY AND INTRATHECALLY ADMINISTERED SCH527123 REVERSED HINDPAW ALLODYNIA AFTER TBI. MOST OF THE SPINAL CXCR2 APPEARED TO BE EXPRESSED BY SPINAL CORD NEURONS. CHROMATIN IMMUNOPRECIPITATION EXPERIMENTS DEMONSTRATED TBI-ENHANCED ASSOCIATION OF THE CXCR2 PROMOTER WITH ACETYLATED-H3K9 HISTONE PROTEIN THAT WAS ALSO REVERSIBLE USING ANACARDIC ACID. TAKEN TOGETHER, OUR FINDINGS SUGGESTED THAT TBI CAUSES THE UPREGULATION OF SPINAL CXCR2 THROUGH AN EPIGENETIC MECHANISM ULTIMATELY SUPPORTING NOCICEPTIVE SENSITIZATION. THE USE OF CXCR2 ANTAGONISTS MAY, THEREFORE, BE USEFUL IN PAIN RESULTING FROM TBI.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
11  5752  28 SOCIAL ISOLATION AND SOCIAL SUPPORT AT ADULTHOOD AFFECT EPIGENETIC MECHANISMS, BRAIN-DERIVED NEUROTROPHIC FACTOR LEVELS AND BEHAVIOR OF CHRONICALLY STRESSED RATS. EPIGENETIC MODULATION OF BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) PROVIDES ONE POSSIBLE EXPLANATION FOR THE DYSFUNCTIONS INDUCED BY STRESS, SUCH AS PSYCHIATRIC DISORDERS AND COGNITIVE DECLINE. INTERESTINGLY, SOCIAL SUPPORT CAN BE PROTECTIVE AGAINST SOME OF THESE EFFECTS, BUT THE MECHANISMS OF SOCIAL BUFFERING ARE POORLY UNDERSTOOD. CONVERSELY, EARLY ISOLATION EXACERBATES THE RESPONSES TO STRESSORS, ALTHOUGH ITS EFFECTS IN ADULTHOOD REMAIN UNCLEAR. THIS STUDY INVESTIGATED THE EFFECTS OF SOCIAL ISOLATION AND SOCIAL BUFFERING ON HIPPOCAMPAL EPIGENETIC MECHANISMS, BDNF LEVELS AND BEHAVIORAL RESPONSES OF CHRONICALLY STRESSED YOUNG ADULT RATS. MALE WISTAR RATS (3 MONTHS) WERE ASSIGNED TO ACCOMPANIED (PAIRED) OR ISOLATED HOUSING. AFTER ONE-MONTH HALF OF EACH GROUP WAS SUBMITTED TO A CHRONIC UNPREDICTABLE STRESS (CUS) PROTOCOL FOR 18 DAYS. AMONG ACCOMPANIED ANIMALS, ONLY ONE WAS EXPOSED TO STRESS. BEHAVIORAL ANALYSIS ENCOMPASSED THE OPEN FIELD, PLUS MAZE AND INHIBITORY AVOIDANCE TASKS. HIPPOCAMPAL H3K9 AND H4K12 ACETYLATION, HDAC5 EXPRESSION AND BDNF LEVELS WERE EVALUATED. ISOLATED HOUSING INCREASED HDAC5 EXPRESSION, DECREASED H3K9 AND H4K12 ACETYLATION, REDUCED BDNF LEVELS, AND IMPAIRED LONG-TERM MEMORY. STRESS AFFECTED WEIGHT GAIN, INDUCED ANXIETY-LIKE BEHAVIOR AND DECREASED ACK9H3 LEVELS. INTERACTIONS BETWEEN HOUSING CONDITIONS AND SOCIAL STRESS WERE SEEN ONLY FOR HDAC5 EXPRESSION, WHICH SHOWED A FURTHER INCREASE IN THE ISOLATED + CUS GROUP BUT REMAINED CONSTANT IN ACCOMPANIED ANIMALS. IN CONCLUSION, SOCIAL ISOLATION AT ADULTHOOD INDUCED EPIGENETIC ALTERATIONS AND EXACERBATED THE EFFECTS OF CHRONIC STRESS ON HDAC5. NOTWITHSTANDING, SOCIAL SUPPORT COUNTERACTED THE ADVERSE EFFECTS OF STRESS ON HDAC5 EXPRESSION.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
12  6418  40 THE TEMPORAL EXPRESSION OF CIRCULATING MICRORNAS AFTER ACUTE EXPERIMENTAL PAIN IN HUMANS. BACKGROUND: MICRORNAS (MIRNAS) CAN MODULATE SEVERAL BIOLOGICAL SYSTEMS, INCLUDING THE PAIN SYSTEM. THIS STUDY AIMED TO EVALUATE THE TEMPORAL EXPRESSION OF CIRCULATING MIRNAS IN THE PLASMA OF HEALTHY VOLUNTEERS AS A MARKER FOR EPIGENETIC CHANGES BEFORE AND AFTER AN ACUTE, EXPERIMENTAL, PAIN PROVOCATION BY INTRAMUSCULAR HYPERTONIC SALINE INJECTION. METHODS: TWENTY VOLUNTEERS WERE RANDOMLY ALLOCATED INTO TWO GROUPS AND RECEIVED EITHER HYPERTONIC (PAIN) OR ISOTONIC (CONTROL) SALINE INJECTION IN THE FIRST DORSAL INTEROSSEOUS MUSCLE OF THEIR DOMINANT HAND. PAIN INTENSITY WAS CONTINUOUSLY RECORDED FOR 20 MINUTES AFTER INJECTION ON A VAS SCALE FROM 0 TO 100 (0 INDICATES NO PAIN AND 100 THE WORST IMAGINABLE PAIN). BLOOD SAMPLES WERE TAKEN AT BASELINE, 30 MINUTES, 3 HOURS, AND 24 HOURS POST-INJECTION, AND PLASMA WAS SEPARATED. MIRNA EXTRACTS WERE USED FOR RNA SEQUENCING WITH THE ILLUMINA NEXTSEQ PLATFORM. MIRNA TRANSCRIPTS WERE COMPARED BETWEEN THE PAIN AND THE NO-PAIN, CONTROL GROUP AT EVERY TIME POINT. SIGNIFICANT DIFFERENCES WERE CONSIDERED WHEN FOLDS WERE >2 AND THE FALSE DISCOVERY RATE WAS P < 0.05. RESULTS: AFTER 30 MINUTES, 4 MIRNAS WERE SIGNIFICANTLY ALTERED IN THE PAIN GROUP COMPARED TO CONTROLS, WHICH INCREASED TO 24 AFTER 3 HOURS AND TO 42 AFTER 24 HOURS FROM BASELINE (P < 0.0001). TWO MIRNAS WERE CONSISTENTLY UPREGULATED THROUGHOUT THE EXPERIMENT. ENRICHMENT ANALYSIS SHOWED SIGNIFICANT MIRNAS INVOLVED IN BRAIN PERCEPTION OF PAIN, BRAIN SIGNALLING AND RESPONSE TO STIMULI. CONCLUSIONS: THIS EXPLORATORY STUDY IS THE FIRST TO REPORT ON THE TEMPORAL EXPRESSION OF CIRCULATING MIRNAS AFTER AN ACUTE, HUMAN EXPERIMENTAL MUSCLE PAIN MODEL. SIGNIFICANCE: THIS EXPLORATORY STUDY EVALUATED THE TEMPORAL PROFILE OF CIRCULATING MIRNAS IN THE PLASMA OF HEALTHY SUBJECTS AFTER ACUTE EXPERIMENTAL PAIN. SEVERAL MIRNAS WERE ALTERED IN SUBJECTS AT THE TIMES OF FOLLOW-UP AFTER THE ACUTE PAIN MODEL WHEN COMPARED TO CONTROLS. MIRNAS PREVIOUSLY ASSOCIATED WITH PAIN PROCESSES WERE ALTERED IN THE PAIN GROUP. OUR RESULTS, BY SHOWING THE FAST AND PROLONGED MODIFICATIONS OF MIRNA ELICITED BY THE ACUTE EXPERIMENTAL PAIN MODEL, ADD NEW PERSPECTIVES TO THE TOPIC OF EPIGENETICS AND PAIN.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
13  6108  31 THE ENRICHED ENVIRONMENT AMELIORATES CHRONIC UNPREDICTABLE MILD STRESS-INDUCED DEPRESSIVE-LIKE BEHAVIORS AND COGNITIVE IMPAIRMENT BY ACTIVATING THE SIRT1/MIR-134 SIGNALING PATHWAY IN HIPPOCAMPUS. BACKGROUND: CHRONIC UNPREDICTABLE MILD STRESS (CUMS) IS AN IMPORTANT RISK FACTOR FOR DEPRESSION AND COGNITIVE DEFICITS IN HUMANS. ENRICHED ENVIRONMENT (EE) SHOWED A BENEFICIAL EFFECT ON DEPRESSION AND COGNITION BY ENHANCING BRAIN DERIVED NEUROTROPHIC FACTOR (BDNF) EXPRESSION AND SYNAPTIC PLASTICITY. HOWEVER, IT IS STILL NOT CLEARLY UNDERSTOOD WHETHER AN EPIGENETIC MECHANISM IS INVOLVED IN THE BDNF MODULATION AND SYNAPTIC PLASTICITY THAT OCCURS AFTER EE TREATMENT FOR THE DEPRESSIVE-LIKE BEHAVIORS AND COGNITIVE DEFICITS ELICITED BY CUMS. IN THIS STUDY, WE INVESTIGATED THE POSSIBLE MECHANISM OF THE NEUROPROTECTIVE EFFECT OF EE. METHODS: ALL RATS WERE EXPOSED TO THE 5-WEEK CUMS PROCEDURE EXCEPT THE CONTROL GROUP. AFTER CUMS PROCEDURE, SOME RATS WERE STEREOTAXICALLY INJECTED WITH SIRT1 PHARMACOLOGIC INHIBITOR EX527 OR SIRT1 KNOCKING DOWN LENTIVIRUS (SH-SIRT1) IN THE HIPPOCAMPUS FOLLOWED BY EE TREATMENT FOR 3 WEEKS. OTHER RATS WERE DIRECTLY SUBJECTED TO EE TREATMENT WITHOUT STEREOTAXIC INJECTION. BEHAVIORAL TESTS WERE USED TO APPRAISE DEPRESSION AND COGNITION AFTER EE TREATMENT. THEN EPIGENETIC MOLECULES, SYNAPTIC PROTEINS, DENDRITIC SPINE DENSITY AND BRANCHES, AND SYNAPTIC MORPHOLOGY OF THE DORSAL HIPPOCAMPUS WERE DETERMINED. RESULTS: WE FOUND THAT CUMS INDUCED DEPRESSIVE-LIKE BEHAVIORS INCLUDING DECREASED SUCROSE PREFERENCE RATIO, PROLONGED IMMOBILITY AND REDUCED LOCOMOTOR AND EXPLORATORY ACTIVITY; COGNITIVE DEFICITS INCLUDING SPATIAL LEARNING AND MEMORY IMPAIRMENT; REDUCED DENDRITIC SPINE DENSITY AND NUMBER OF BRANCHES; THINNED POSTSYNAPTIC DENSITY; DOWNREGULATED SIRT1/MICRORNA-134 PATHWAY, DECREASED BDNF AND SYNAPTIC PROTEINS INCLUDING SYNAPTOPHYSIN (SYN) AND POSTSYNAPTIC DENSITY PROTEIN 95 (PSD95) EXPRESSION IN THE HIPPOCAMPUS. HOWEVER, THE CUMS-INDUCED DEPRESSIVE-LIKE BEHAVIORS, COGNITIVE DEFICITS, DENDRITIC SPINE DENSITY AND BRANCH NUMBER REDUCTION, POSTSYNAPTIC DENSITY THINNING, SIRT1/MICRORNA-134 PATHWAY DOWNREGULATION, BDNF AND SYNAPTIC PROTEINS REDUCTION, INCLUDING SYNAPTOPHYSIN (SYN) AND POSTSYNAPTIC DENSITY PROTEIN 95 (PSD95), WERE REVERSED BY EE TREATMENT. HOWEVER, DEPRESSIVE-LIKE BEHAVIORS AND COGNITIVE DEFICITS WERE OBSERVED AGAIN IN RATS SUBJECTED TO STEREOTAXIC INJECTION WITH EX527 OR SH-SIRT1. FURTHERMORE, THIS STUDY ALSO FOUND THAT SIRT1/MICRORNA-134 REGULATES THE DOWNSTREAM MOLECULES BDNF, AND THE SYNAPTIC PROTEINS SYN AND PSD95 IN PRIMARY CULTURED HIPPOCAMPAL NEURONS. CONCLUSIONS: THIS STUDY PROVIDES EVIDENCE FOR THE NEUROPROTECTIVE ROLE OF EE ON DEPRESSION AND COGNITIVE DEFICITS BY ACTIVATING THE SIRT1/MICRORNA-134 PATHWAY, WHICH ACCOUNTS FOR THE REGULATION OF SYNAPTIC PROTEINS, INCLUDING BDNF, PSD95 AND SYN, DENDRITIC REMODELING AND ULTRASTRUCTURE CHANGES OF SYNAPSES IN THE HIPPOCAMPUS.	2019	

14  5760  36 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
15  1875  41 EMERGING ROLE OF ONE-CARBON METABOLISM AND DNA METHYLATION ENRICHMENT ON DELTA-CONTAINING GABAA RECEPTOR EXPRESSION IN THE CEREBELLUM OF SUBJECTS WITH ALCOHOL USE DISORDERS (AUD). BACKGROUND: CEREBELLUM IS AN AREA OF THE BRAIN PARTICULARLY SENSITIVE TO THE EFFECTS OF ACUTE AND CHRONIC ALCOHOL CONSUMPTION. ALCOHOL EXPOSURE DECREASES CEREBELLAR PURKINJE CELL OUTPUT BY INCREASING GABA RELEASE FROM GOLGI CELLS ONTO EXTRASYNAPTIC ALPHA6/DELTA-CONTAINING GABAA RECEPTORS LOCATED ON GLUTAMATERGIC GRANULE CELLS. HERE, WE STUDIED WHETHER CHRONIC ALCOHOL CONSUMPTION INDUCES CHANGES IN GABAA RECEPTOR SUBUNIT EXPRESSION AND WHETHER THESE CHANGES ARE ASSOCIATED WITH ALTERATIONS IN EPIGENETIC MECHANISMS VIA DNA METHYLATION. METHODS: WE USED A COHORT OF POSTMORTEM CEREBELLUM FROM CONTROL AND CHRONIC ALCOHOLICS, HERE DEFINED AS ALCOHOL USE DISORDERS SUBJECTS (N=25/GROUP). S-ADENOSYL-METHIONINE/S-ADENOSYL-HOMOCYSTEINE WERE MEASURED BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY. MRNA LEVELS OF VARIOUS GENES WERE ASSESSED BY REVERSE TRANSCRIPTASE-QUANTITATIVE POLYMERASE CHAIN REACTION. PROMOTER METHYLATION ENRICHMENT WAS ASSESSED USING METHYLATED DNA IMMUNOPRECIPITATION AND HYDROXY-METHYLATED DNA IMMUNOPRECIPITATION ASSAYS. RESULTS: MRNAS ENCODING KEY ENZYMES OF 1-CARBON METABOLISM THAT DETERMINE THE S-ADENOSYL-METHIONINE/S-ADENOSYL-HOMOCYSTEINE RATIO WERE INCREASED, INDICATING HIGHER "METHYLATION INDEX" IN ALCOHOL USE DISORDER SUBJECTS. WE FOUND THAT INCREASED METHYLATION OF THE PROMOTER OF THE DELTA SUBUNIT GABAA RECEPTOR WAS ASSOCIATED WITH REDUCED MRNA AND PROTEIN LEVELS IN THE CEREBELLUM OF ALCOHOL USE DISORDER SUBJECTS. NO CHANGES WERE OBSERVED IN ALPHA1- OR ALPHA6-CONTAINING GABAA RECEPTOR SUBUNITS. THE EXPRESSION OF DNA-METHYLTRANSFERASES (1, 3A, AND 3B) WAS UNALTERED, WHEREAS THE MRNA LEVEL OF TET1, WHICH PARTICIPATES IN THE DNA DEMETHYLATION PATHWAY, WAS DECREASED. HENCE, INCREASED METHYLATION OF THE DELTA SUBUNIT GABAA RECEPTOR PROMOTER MAY RESULT FROM ALCOHOL-INDUCED REDUCTION OF DNA DEMETHYLATION. CONCLUSION: TOGETHER, THESE RESULTS SUPPORT THE HYPOTHESIS THAT ABERRANT DNA METHYLATION PATHWAYS MAY BE INVOLVED IN CEREBELLAR PATHOPHYSIOLOGY OF ALCOHOLISM. FURTHERMORE, THIS WORK PROVIDES NOVEL EVIDENCE FOR A CENTRAL ROLE OF DNA METHYLATION MECHANISMS IN THE ALCOHOL-INDUCED NEUROADAPTIVE CHANGES OF HUMAN CEREBELLAR GABAA RECEPTOR FUNCTION.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
16   169  38 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
17  2827  25 FLUOXETINE INCREASES HIPPOCAMPAL NEUROGENESIS AND INDUCES EPIGENETIC FACTORS BUT DOES NOT IMPROVE FUNCTIONAL RECOVERY AFTER TRAUMATIC BRAIN INJURY. THE SELECTIVE SEROTONIN REUPTAKE INHIBITOR FLUOXETINE INDUCES HIPPOCAMPAL NEUROGENESIS, STIMULATES MATURATION AND SYNAPTIC PLASTICITY OF ADULT HIPPOCAMPAL NEURONS, AND REDUCES MOTOR/SENSORY AND MEMORY IMPAIRMENTS IN SEVERAL CNS DISORDERS. IN THE SETTING OF TRAUMATIC BRAIN INJURY (TBI), ITS EFFECTS ON NEUROPLASTICITY AND FUNCTION HAVE YET TO BE THOROUGHLY INVESTIGATED. HERE WE EXAMINED THE EFFICACY OF FLUOXETINE AFTER A MODERATE TO SEVERE TBI, PRODUCED BY A CONTROLLED CORTICAL IMPACT. THREE DAYS AFTER TBI OR SHAM SURGERY, MICE WERE TREATED WITH FLUOXETINE (10 MG/KG/D) OR VEHICLE FOR 4 WEEKS. TO EVALUATE THE EFFECTS OF FLUOXETINE ON NEUROPLASTICITY, HIPPOCAMPAL NEUROGENESIS AND EPIGENETIC MODIFICATION WERE STUDIED. STEREOLOGIC ANALYSIS OF THE DENTATE GYRUS REVEALED A SIGNIFICANT INCREASE IN DOUBLECORTIN-POSITIVE CELLS IN BRAIN-INJURED ANIMALS TREATED WITH FLUOXETINE RELATIVE TO CONTROLS, A FINDING CONSISTENT WITH ENHANCED HIPPOCAMPAL NEUROGENESIS. EPIGENETIC MODIFICATIONS, INCLUDING AN INCREASE IN HISTONE 3 ACETYLATION AND INDUCTION OF METHYL-CPG-BINDING PROTEIN, A TRANSCRIPTION FACTOR INVOLVED IN DNA METHYLATION, WERE LIKEWISE SEEN BY IMMUNOHISTOCHEMISTRY AND QUANTITATIVE WESTERN IMMUNOBLOTS, RESPECTIVELY, IN BRAIN-INJURED ANIMALS TREATED WITH FLUOXETINE. TO DETERMINE IF FLUOXETINE IMPROVES NEUROLOGICAL OUTCOMES AFTER TBI, GAIT FUNCTION AND SPATIAL LEARNING AND MEMORY WERE ASSESSED BY THE CATWALK-ASSISTED GAIT TEST AND BARNES MAZE TEST, RESPECTIVELY. NO DIFFERENCES IN THESE PARAMETERS WERE SEEN BETWEEN FLUOXETINE- AND VEHICLE-TREATED ANIMALS. THUS WHILE FLUOXETINE ENHANCED NEUROPLASTICITY IN THE HIPPOCAMPUS AFTER TBI, ITS CHRONIC ADMINISTRATION DID NOT RESTORE LOCOMOTOR FUNCTION OR AMELIORATE MEMORY DEFICITS.	2011	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
18  1026  39 CIRCULATING MIRNAS EXPRESSION IN MYALGIC ENCEPHALOMYELITIS/CHRONIC FATIGUE SYNDROME. MYALGIC ENCEPHALOMYELITIS/CHRONIC FATIGUE SYNDROME (ME/CFS) IS A COMPLEX MULTIFACTORIAL DISEASE THAT CAUSES INCREASING MORBIDITY WORLDWIDE, AND MANY INDIVIDUALS WITH ME/CFS SYMPTOMS REMAIN UNDIAGNOSED DUE TO THE LACK OF DIAGNOSTIC BIOMARKERS. ITS ETIOLOGY IS STILL UNKNOWN, BUT INCREASING EVIDENCE SUPPORTS A ROLE OF HERPESVIRUSES (INCLUDING HHV-6A AND HHV-6B) AS POTENTIAL TRIGGERS. INTERESTINGLY, THE INFECTION BY THESE VIRUSES HAS BEEN REPORTED TO IMPACT THE EXPRESSION OF MICRORNAS (MIRNAS), SHORT NON-CODING RNA SEQUENCES WHICH HAVE BEEN SUGGESTED TO BE EPIGENETIC FACTORS MODULATING ME/CFS PATHOGENIC MECHANISMS. NOTABLY, THE PRESENCE OF CIRCULATING MIRNAS IN PLASMA HAS RAISED THE POSSIBILITY TO USE THEM AS VALUABLE BIOMARKERS FOR DISTINGUISHING ME/CFS PATIENTS FROM HEALTHY CONTROLS. THUS, THIS STUDY AIMED AT DETERMINING THE ROLE OF EIGHT MIRNAS, WHICH WERE SELECTED FOR THEIR PREVIOUS ASSOCIATION WITH ME/CFS, AS POTENTIAL CIRCULATING BIOMARKERS OF THE DISEASE. THEIR PRESENCE WAS QUANTITATIVELY EVALUATED IN PLASMA FROM 40 ME/CFS PATIENTS AND 20 HEALTHY CONTROLS BY SPECIFIC TAQMAN ASSAYS, AND THE RESULTS SHOWED THAT SIX OUT OF THE EIGHT OF THE SELECTED MIRNAS WERE DIFFERENTLY EXPRESSED IN PATIENTS COMPARED TO CONTROLS; MORE SPECIFICALLY, FIVE MIRNAS WERE SIGNIFICANTLY UPREGULATED (MIR-127-3P, MIR-142-5P, MIR-143-3P, MIR-150-5P, AND MIR-448), AND ONE WAS DOWNMODULATED (MIR-140-5P). MIRNA LEVELS DIRECTLY CORRELATED WITH DISEASE SEVERITY, WHEREAS NO SIGNIFICANT CORRELATIONS WERE OBSERVED WITH THE PLASMA LEVELS OF SEVEN PRO-INFLAMMATORY CYTOKINES OR WITH THE PRESENCE/LOAD OF HHV-6A/6B GENOME, AS JUDGED BY SPECIFIC PCR AMPLIFICATION. THE RESULTS MAY OPEN THE WAY FOR FURTHER VALIDATION OF MIRNAS AS NEW POTENTIAL BIOMARKERS IN ME/CFS AND INCREASE THE KNOWLEDGE OF THE COMPLEX PATHWAYS INVOLVED IN THE ME/CFS DEVELOPMENT.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
19  4647  38 NEUROPEPTIDE AND SMALL TRANSMITTER COEXISTENCE: FUNDAMENTAL STUDIES AND RELEVANCE TO MENTAL ILLNESS. NEUROPEPTIDES ARE AUXILIARY MESSENGER MOLECULES THAT ALWAYS CO-EXIST IN NERVE CELLS WITH ONE OR MORE SMALL MOLECULE (CLASSIC) NEUROTRANSMITTERS. NEUROPEPTIDES ACT BOTH AS TRANSMITTERS AND TROPHIC FACTORS, AND PLAY A ROLE PARTICULARLY WHEN THE NERVOUS SYSTEM IS CHALLENGED, AS BY INJURY, PAIN OR STRESS. HERE NEUROPEPTIDES AND COEXISTENCE IN MAMMALS ARE REVIEWED, BUT WITH SPECIAL FOCUS ON THE 29/30 AMINO ACID GALANIN AND ITS THREE RECEPTORS GALR1, -R2 AND -R3. IN PARTICULAR, GALANIN'S ROLE AS A CO-TRANSMITTER IN BOTH RODENT AND HUMAN NORADRENERGIC LOCUS COERULEUS (LC) NEURONS IS ADDRESSED. EXTENSIVE EXPERIMENTAL ANIMAL DATA STRONGLY SUGGEST A ROLE FOR THE GALANIN SYSTEM IN DEPRESSION-LIKE BEHAVIOR. THE TRANSLATIONAL POTENTIAL OF THESE RESULTS WAS TESTED BY STUDYING THE GALANIN SYSTEM IN POSTMORTEM HUMAN BRAINS, FIRST IN NORMAL BRAINS, AND THEN IN A COMPARISON OF FIVE REGIONS OF BRAINS OBTAINED FROM DEPRESSED PEOPLE WHO COMMITTED SUICIDE, AND FROM MATCHED CONTROLS. THE DISTRIBUTION OF GALANIN AND THE FOUR GALANIN SYSTEM TRANSCRIPTS IN THE NORMAL HUMAN BRAIN WAS DETERMINED, AND SELECTIVE AND PARALLEL CHANGES IN LEVELS OF TRANSCRIPTS AND DNA METHYLATION FOR GALANIN AND ITS THREE RECEPTORS WERE ASSESSED IN DEPRESSED PATIENTS WHO COMMITTED SUICIDE: UPREGULATION OF TRANSCRIPTS, E.G., FOR GALANIN AND GALR3 IN LC, PARALLELED BY A DECREASE IN DNA METHYLATION, SUGGESTING INVOLVEMENT OF EPIGENETIC MECHANISMS. IT IS HYPOTHESIZED THAT, WHEN EXPOSED TO SEVERE STRESS, THE NORADRENERGIC LC NEURONS FIRE IN BURSTS AND RELEASE GALANIN FROM THEIR SOMA/DENDRITES. GALANIN THEN ACTS ON SOMATO-DENDRITIC, INHIBITORY GALANIN AUTORECEPTORS, OPENING POTASSIUM CHANNELS AND INHIBITING FIRING. THE PURPOSE OF THESE AUTORECEPTORS IS TO ACT AS A 'BRAKE' TO PREVENT OVEREXCITATION, A BRAKE THAT IS ALSO PART OF RESILIENCE TO STRESS THAT PROTECTS AGAINST DEPRESSION. DEPRESSION THEN ARISES WHEN THE INHIBITION IS TOO STRONG AND LONG LASTING - A MALADAPTION, ALLOSTATIC LOAD, LEADING TO DEPLETION OF NA LEVELS IN THE FOREBRAIN. IT IS SUGGESTED THAT DISINHIBITION BY A GALANIN ANTAGONIST MAY HAVE ANTIDEPRESSANT ACTIVITY BY RESTORING FOREBRAIN NA LEVELS. A ROLE OF GALANIN IN DEPRESSION IS ALSO SUPPORTED BY A RECENT CANDIDATE GENE STUDY, SHOWING THAT VARIANTS IN GENES FOR GALANIN AND ITS THREE RECEPTORS CONFER INCREASED RISK OF DEPRESSION AND ANXIETY IN PEOPLE WHO EXPERIENCED CHILDHOOD ADVERSITY OR RECENT NEGATIVE LIFE EVENTS. IN SUMMARY, GALANIN, A NEUROPEPTIDE COEXISTING IN LC NEURONS, MAY PARTICIPATE IN THE MECHANISM UNDERLYING RESILIENCE AGAINST A SERIOUS AND COMMON DISORDER, MDD. EXISTING AND FURTHER RESULTS MAY LEAD TO AN INCREASED UNDERSTANDING OF HOW THIS ILLNESS DEVELOPS, WHICH IN TURN COULD PROVIDE A BASIS FOR ITS TREATMENT.	2018	
                                                                                                                  
20  4275  28 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