Search results for " Methods"

showing 10 items of 4102 documents

Increased PD-1 Expression and Altered T Cell Repertoire Diversity Predict Mortality in Patients with Septic Shock: A Preliminary Study

2017

Sepsis causes impairment of innate and adaptive immunity by multiple mechanisms, including depletion of immune effector cells and T cell exhaustion. Although lymphocyte dysfunction is associated with increased mortality and potential reactivation of latent viral infection in patients with septic shock, the relation between viral reactivation and lymphocyte dysfunction is obscure. The objectives of this study were 1) to determine the relation of lymphocyte dysfunction to viral reactivation and mortality, and 2) to evaluate recovery of lymphocyte function during septic shock, including T cell receptor (TCR) diversity and the expression of programmed death 1 (PD-1). In 18 patients with septic …

0301 basic medicineMaleLymphocyteReceptor expressionProgrammed Cell Death 1 Receptorlcsh:MedicineCytomegalovirusGene ExpressionArtificial Gene Amplification and ExtensionPathology and Laboratory MedicineImmune ReceptorsBiochemistryPolymerase Chain ReactionMonocytesWhite Blood Cells0302 clinical medicineSpectrum Analysis TechniquesAnimal CellsT-Lymphocyte SubsetsMedicine and Health SciencesLymphocyteslcsh:ScienceAged 80 and overMultidisciplinaryImmune System ProteinsT CellsMiddle AgedAcquired immune systemFlow CytometryPrognosisShock Septicmedicine.anatomical_structurePhenotypeSpectrophotometryShock (circulatory)Cytomegalovirus InfectionsFemaleCytophotometrymedicine.symptomCellular TypesResearch ArticleSignal TransductionT cellImmune CellsImmunologyReceptors Antigen T-CellBiologyResearch and Analysis MethodsMicrobiologyImmunophenotypingSepsis03 medical and health sciencesImmune systemSigns and SymptomsDiagnostic MedicineSepsisVirologymedicineHumansMolecular Biology TechniquesMolecular BiologyAgedBlood CellsSeptic shocklcsh:RBiology and Life SciencesProteins030208 emergency & critical care medicineCell BiologyHLA-DR Antigensmedicine.diseaseViral ReplicationT Cell Receptors030104 developmental biologyCase-Control StudiesImmunologylcsh:QBiomarkersPLoS ONE
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Use of Cepheid Xpert Carba-R® for Rapid Detection of Carbapenemase-Producing Bacteria in Abdominal Septic Patients Admitted to Intensive Care Unit.

2016

Abstract Early institution of effective antibiotic therapy and source control are pivotal to improve survival of abdominal septic patients. Xpert® Carba-R is a real time polymerase chain reaction assay for rapid detection and differentiation of five genes (blaKPC, blaVIM, blaOXA-48, blaIMP-1, blaNDM) responsible for carbapenem resistance. We performed an observational study investigating the clinical usefulness and applicability of Xpert® Carba-R to detect carbapenem resistance in abdominal septic patients admitted to intensive care unit. We compared the results of Xpert® Carba-R with standard microbiological culture. We collected a set of two rectal/stomia swabs and two swabs from abdomina…

0301 basic medicineMaleMicrobiological cultureAntibioticslcsh:MedicineArtificial Gene Amplification and ExtensionPathology and Laboratory MedicinePolymerase Chain Reactionlaw.inventionKlebsiella Pneumoniae0302 clinical medicinelawAntibioticsKlebsiellaEpidemiologymultidrug resistance sepsis intensive care unitAbdomenMedicine and Health SciencesMedicine030212 general & internal medicinelcsh:ScienceMultidisciplinaryAntimicrobialsCepheid Xpert Carba-R®DrugsMicrobial CulturesMiddle AgedIntensive care unitHospitalsBacterial PathogensIntensive Care UnitsAbdominal SurgeryMedical MicrobiologyFemaleBiological CulturesPathogensResearch ArticleDNA Bacterialmedicine.medical_specialtymedicine.drug_class030106 microbiologySurgical and Invasive Medical ProceduresResearch and Analysis MethodsReal-Time Polymerase Chain ReactionRapid detectionMicrobiologySensitivity and Specificitybeta-Lactamases03 medical and health sciencesAntibiotic resistanceBacterial ProteinsEnterobacteriaceaeDiagnostic MedicineInternal medicineIntensive careMicrobial ControlSepsisDrug Resistance BacterialHumansMED/41 - ANESTESIOLOGIAMolecular Biology TechniquesMicrobial PathogensMolecular BiologyAgedPharmacologyBacteriabusiness.industrylcsh:ROrganismsRectumBiology and Life SciencesSurgeryHealth CareCarbapenemsHealth Care FacilitiesAntibiotic Resistancelcsh:QAntimicrobial ResistanceReagent Kits DiagnosticbusinessAbdominal surgery
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Odiparcil, a potential glycosaminoglycans clearance therapy in mucopolysaccharidosis VI—Evidence from in vitro and in vivo models

2020

International audience; Mucopolysaccharidoses are a class of lysosomal storage diseases, characterized by enzymatic deficiency in the degradation of specific glycosaminoglycans (GAG). Pathological accumulation of excess GAG leads to multiple clinical symptoms with systemic character, most severely affecting bones, muscles and connective tissues. Current therapies include periodic intravenous infusion of supplementary recombinant enzyme (Enzyme Replacement Therapy-ERT) or bone marrow transplantation. However, ERT has limited efficacy due to poor penetration in some organs and tissues. Here, we investigated the potential of the β-D-xyloside derivative odiparcil as an oral GAG clearance therap…

0301 basic medicineMaleMucopolysaccharidosis type VIRespiratory SystemAdministration OralGlycosaminoglycanRats Sprague-DawleyWhite Blood CellsMice0302 clinical medicineOral administrationAnimal CellsMedicine and Health SciencesGlycosidesCells CulturedConnective Tissue CellsGlycosaminoglycansMultidisciplinaryMucopolysaccharidosis VIChemistryChondroitin SulfatesQRMucopolysaccharidosis VIAnimal Models3. Good healthTracheamedicine.anatomical_structureExperimental Organism SystemsConnective Tissue[SDV.SP.PHARMA] Life Sciences [q-bio]/Pharmaceutical sciences/PharmacologyMedicineFemaleBiological CulturesCellular TypesAnatomyCellular Structures and OrganellesResearch Articlemedicine.medical_specialtyImmune CellsScienceImmunologyDermatan SulfateMouse ModelsIn Vitro TechniquesResearch and Analysis Methods03 medical and health sciencesModel OrganismsIn vivoInternal medicinemedicineAnimalsHumansBlood CellsCartilageBiology and Life SciencesEndothelial CellsKidneysCell BiologyRenal SystemFibroblastsCell CulturesIn vitroMice Mutant StrainsRatsMice Inbred C57BLDisease Models Animal030104 developmental biologyEndocrinologyBiological TissueCartilageCell cultureAnimal Studies[SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/PharmacologyCattleLysosomes030217 neurology & neurosurgery
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Oligodendrocytes support axonal transport and maintenance via exosome secretion

2020

Neurons extend long axons that require maintenance and are susceptible to degeneration. Long-term integrity of axons depends on intrinsic mechanisms including axonal transport and extrinsic support from adjacent glial cells. The mechanisms of support provided by myelinating oligodendrocytes to underlying axons are only partly understood. Oligodendrocytes release extracellular vesicles (EVs) with properties of exosomes, which upon delivery to neurons improve neuronal viability in vitro. Here, we show that oligodendroglial exosome secretion is impaired in 2 mouse mutants exhibiting secondary axonal degeneration due to oligodendrocyte-specific gene defects. Wild-type oligodendroglial exosomes …

0301 basic medicineMaleMutantHippocampusCentrifugationExosomesAxonal TransportHippocampusMass SpectrometryAnalytical ChemistryMiceMyelin0302 clinical medicineNerve FibersSpectrum Analysis TechniquesAnimal CellsMedicine and Health SciencesBiology (General)Myelin SheathNeuronsLiquid ChromatographyGeneral NeuroscienceChromatographic TechniquesBrainCell biologyChemistrySeparation ProcessesOligodendrogliamedicine.anatomical_structureCell ProcessesPhysical SciencesFemaleCellular TypesCellular Structures and OrganellesAnatomyGeneral Agricultural and Biological SciencesNeurogliaResearch ArticleSignal TransductionMaintenanceQH301-705.5Liquid Chromatography-Mass SpectrometryBiologyResearch and Analysis MethodsExosomeGeneral Biochemistry Genetics and Molecular Biology03 medical and health sciencesExtracellular VesiclesmedicineAnimalsHumansSecretionVesiclesGeneral Immunology and MicrobiologyWild typeBiology and Life SciencesCell BiologyIn vitroAxonsMicrovesiclesMice Inbred C57BL030104 developmental biologyHEK293 Cellsnervous systemCellular NeuroscienceAxoplasmic transportNeuronUltracentrifugation030217 neurology & neurosurgeryNeuroscience
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Different behavior of myeloperoxidase in two rodent amoebic liver abscess models.

2016

The protozoan Entamoeba histolytica is the etiological agent of amoebiasis, which can spread to the liver and form amoebic liver abscesses. Histological studies conducted with resistant and susceptible models of amoebic liver abscesses (ALAs) have established that neutrophils are the first cells to contact invasive amoebae at the lesion site. Myeloperoxidase is the most abundant enzyme secreted by neutrophils. It uses hydrogen peroxide secreted by the same cells to oxidize chloride ions and produce hypochlorous acid, which is the most efficient microbicidal system of neutrophils. In a previous report, our group demonstrated that myeloperoxidase presents amoebicidal activity in vitro. The ai…

0301 basic medicineMalePathologyNeutrophilslcsh:MedicineGene ExpressionPathology and Laboratory MedicineWhite Blood Cells0302 clinical medicineAnimal CellsCricetinaeMedicine and Health SciencesAmoebaslcsh:ScienceImmune ResponseDisease ResistanceMammalsProtozoansMice Inbred BALB CMultidisciplinaryAmoebic liver abscessbiologyChemistryAnimal ModelsLiverExperimental Organism SystemsMyeloperoxidaseHost-Pathogen InteractionsVertebratesLiver Abscess AmebicHamstersmedicine.symptomCellular TypesResearch Articlemedicine.medical_specialtyImmune CellsImmunologyMouse ModelsResearch and Analysis MethodsRodentsMicrobiologyLesionEntamoeba Histolytica03 medical and health sciencesEntamoeba histolyticaModel OrganismsSigns and SymptomsIn vivoDiagnostic MedicineParasite GroupsmedicineGeneticsAnimalsAmoebiasisTrophozoitesPeroxidaseInflammationBlood Cellslcsh:ROrganismsBiology and Life SciencesCell Biologybiology.organism_classificationmedicine.diseaseIn vitroParasitic ProtozoansDisease Models Animal030104 developmental biologyAmniotesbiology.proteinlcsh:QParasitologyLeukocyte ElastaseApicomplexa030215 immunologyLiver abscessPloS one
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Antibody trapping: A novel mechanism of parasite immune evasion by the trematode Echinostoma caproni

2017

Background Helminth infections are among the most prevalent neglected tropical diseases, causing an enormous impact in global health and the socioeconomic growth of developing countries. In this context, the study of helminth biology, with emphasis on host-parasite interactions, appears as a promising approach for developing new tools to prevent and control these infections. Methods/Principal findings The role that antibody responses have on helminth infections is still not well understood. To go in depth into this issue, work on the intestinal helminth Echinostoma caproni (Trematoda: Echinostomatidae) has been undertaken. Adult parasites were recovered from infected mice and cultured in vi…

0301 basic medicineMalePhysiologyAntibody ResponsePathogenesisPathology and Laboratory MedicineBiochemistryMiceImmune PhysiologyEchinostomaMedicine and Health SciencesParasite hostingEnzyme-Linked ImmunoassaysMicroscopy ImmunoelectronImmune ResponseEchinostomiasisImmune System Proteinsbiologylcsh:Public aspects of medicineProteases030108 mycology & parasitologyEnzymesInfectious DiseasesHelminth InfectionsHost-Pathogen InteractionsTrematodaAntibodyEchinostomaCellular Structures and OrganellesResearch ArticleProtein BindingProteaseslcsh:Arctic medicine. Tropical medicinelcsh:RC955-962ImmunologyAntibodies HelminthContext (language use)Research and Analysis MethodsAntibodies03 medical and health sciencesImmune systemParasitic DiseasesAnimalsSecretionVesiclesImmunoassaysImmune EvasionPublic Health Environmental and Occupational HealthBiology and Life SciencesProteinslcsh:RA1-1270Cell Biologybiology.organism_classificationVirologyDisease Models Animal030104 developmental biologyMicroscopy FluorescenceProteolysisbiology.proteinImmunologic TechniquesEnzymologyPLoS Neglected Tropical Diseases
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High Throughput Sequencing Identifies Misregulated Genes in the Drosophila Polypyrimidine Tract-Binding Protein (hephaestus) Mutant Defective in Sper…

2015

The Drosophila polypyrimidine tract-binding protein (dmPTB or hephaestus) plays an important role during spermatogenesis. The heph2 mutation in this gene results in a specific defect in spermatogenesis, causing aberrant spermatid individualization and male sterility. However, the array of molecular defects in the mutant remains uncharacterized. Using an unbiased high throughput sequencing approach, we have identified transcripts that are misregulated in this mutant. Aberrant transcripts show altered expression levels, exon skipping, and alternative 5' ends. We independently verified these findings by reverse-transcription and polymerase chain reaction (RT-PCR) analysis. Our analysis shows m…

0301 basic medicineMalePhysiologyMutantGene Expressionlcsh:MedicineArtificial Gene Amplification and ExtensionPolymerase Chain ReactionBiochemistryConserved sequence0302 clinical medicineSequencing techniquesReproductive PhysiologyAnimal CellsInvertebrate GenomicsMedicine and Health SciencesDrosophila ProteinsProtein IsoformsCell Cycle and Cell Divisionlcsh:ScienceConserved SequencePhylogenyGeneticsRegulation of gene expressionMultidisciplinarybiologyChromosome BiologyDrosophila MelanogasterMessenger RNAHigh-Throughput Nucleotide SequencingRNA sequencingAnimal ModelsGenomicsSpermatidsInsectsNucleic acidsMeiosisCell ProcessesDrosophilaDrosophila melanogasterTranscription Initiation SiteCellular TypesDrosophila ProteinPolypyrimidine Tract-Binding ProteinResearch ArticleArthropodaMolecular Sequence DataReal-Time Polymerase Chain ReactionResearch and Analysis Methods03 medical and health sciencesModel OrganismsGeneticsAnimalsPolypyrimidine tract-binding proteinRNA MessengerSpermatogenesisMolecular Biology TechniquesMolecular BiologyBinding SitesBase SequenceGene Expression Profilinglcsh:ROrganismsBiology and Life SciencesCell BiologyReverse Transcriptase-Polymerase Chain Reactionbiology.organism_classificationInvertebratesExon skippingSpermGene expression profiling030104 developmental biologyGene OntologyGerm CellsGene Expression RegulationAnimal GenomicsMutationbiology.proteinRNAlcsh:QTranscriptome030217 neurology & neurosurgeryPLoS ONE
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Lamivudine/Adefovir Treatment Increases the Rate of Spontaneous Mutation of Hepatitis B Virus in Patients.

2016

The high levels of genetic diversity shown by hepatitis B virus (HBV) are commonly attributed to the low fidelity of its polymerase. However, the rate of spontaneous mutation of human HBV in vivo is currently unknown. Here, based on the evolutionary principle that the population frequency of lethal mutations equals the rate at which they are produced, we have estimated the mutation rate of HBV in vivo by scoring premature stop codons in 621 publicly available, full-length, molecular clone sequences derived from patients. This yielded an estimate of 8.7 × 10-5 spontaneous mutations per nucleotide per cell infection in untreated patients, which should be taken as an upper limit estimate becau…

0301 basic medicineMaleRNA virusesMutation ratelcsh:Medicinemedicine.disease_causeBiochemistryPolymerasesAdefovirFrameshift Mutationlcsh:SciencePathology and laboratory medicineeducation.field_of_studyMultidisciplinaryMicrobial MutationLamivudineMedical microbiologyResistance mutation3. Good healthLamivudineVirusesFemalePathogensSequence AnalysisImmunosuppressive Agentsmedicine.drugResearch ArticleHepatitis B virusSubstitution MutationPopulationOrganophosphonatesBiologyResearch and Analysis MethodsPolymorphism Single NucleotideMicrobiologyFrameshift mutation03 medical and health sciencesHepatitis B ChronicDrug Resistance ViralDNA-binding proteinsmedicineGeneticsHumanseducationMolecular Biology TechniquesSequencing TechniquesMolecular BiologyHepatitis B virusMedicine and health sciencesPoint mutationAdeninelcsh:RViral pathogensOrganismsBiology and Life SciencesProteinsVirologyMolecular biologyHepatitis virusesMicrobial pathogens030104 developmental biologyMutationlcsh:QCloningPLoS ONE
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Case-based surveillance of measles in Sicily during 2012-2017: The changing molecular epidemiology and implications for vaccine strategies.

2018

Following the indication of the World Health Organization, a national plan for the elimination of measles was approved in Italy and this included the improvement of the molecular surveil- lance of measles viruses and the interruption of indigenous transmission of the disease. Nevertheless, large outbreaks continue to occur in almost all regions of the country, includ- ing Sicily. Here we describe the epidemiology and molecular dynamics of measles viruses as a result of the measles surveillance activity carried out by the “Reference Laboratory for Measles and Rubella” in Sicily over a 5-year period. Biological samples of 259 suspected measles cases were tested for viral RNA detection and a t…

0301 basic medicineMaleRNA virusesViral DiseasesHeredityMeasles Surveillance Vaccine Epidemiology Molecular epidemiology Genotyping Sicily Italylcsh:MedicineSettore MED/42 - Igiene Generale E ApplicataPathology and Laboratory MedicineGeographical locationsEpidemiologyMedicine and Health SciencesPublic and Occupational HealthChildlcsh:ScienceSicilyData ManagementMolecular EpidemiologyMultidisciplinarybiologyTransmission (medicine)Database and informatics methodsSequence analysisPhylogenetic AnalysisVaccination and ImmunizationPhylogeneticsEuropeGenetic MappingInfectious DiseasesItalyMedical MicrobiologyChild PreschoolViral PathogensVirusesRNA ViralPathogensResearch ArticleAdultmedicine.medical_specialtyComputer and Information SciencesAdolescentBioinformaticsMeasles VaccineImmunologyNucleotide SequencingMeasles VirusVariant GenotypesRubellaMeaslesMicrobiologyMeasles virus03 medical and health sciencesYoung AdultmedicineGeneticsHumansEvolutionary SystematicsEuropean UnionMolecular Biology TechniquesSequencing TechniquesGenotypingMicrobial PathogensMolecular BiologyDNA sequence analysisRetrospective StudiesTaxonomyEvolutionary BiologyMolecular epidemiologyBiology and life scienceslcsh:ROrganismsOutbreakInfantbiology.organism_classificationmedicine.diseaseVirologyResearch and analysis methods030104 developmental biologyParamyxoviruseslcsh:QPreventive MedicinePeople and placesMeaslesPLoS ONE
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Longitudinal Analysis of Serum Autoantibody-Reactivities in Patients with Primary Open Angle Glaucoma and Optic Disc Hemorrhage.

2015

Background The aim of our current investigation was to analyze the autoantibody-reactivities of primary open angle glaucoma patients with optic disc hemorrhage as possibly correlated to disease progression by means of a protein microarray approach. Methods Sera of patients with primary open angle glaucoma and optic disc hemorrhage (n = 16) were collected directly after study inclusion (0 weeks) and after 2 weeks, 4 weeks and 12 weeks. As a control group patients with primary open angle glaucoma (n = 18) were used (0 weeks and 12 weeks). Microarrays were incubated and occurring antibody-antigen-reactions were visualized with fluorescence labeled anti-human-IgG secondary antibodies. To detect…

0301 basic medicineMaleRetinal Ganglion CellsSerum ProteinsVisual acuitygenetic structuresEye DiseasesMicroarraysVisionVisual AcuityGlaucomalcsh:MedicineSocial SciencesPathogenesisPathology and Laboratory MedicineVascular MedicineBiochemistryPathogenesis0302 clinical medicineAnimal CellsMedicine and Health SciencesPsychologyLongitudinal Studieslcsh:ScienceNeuronsMultidisciplinarybiologyMiddle AgedPrimary and secondary antibodiesmedicine.anatomical_structureBioassays and Physiological AnalysisDisease ProgressionFemaleSensory PerceptionAntibodymedicine.symptomCellular TypesAnatomyGlaucoma Open-AngleOptic discResearch Articlemedicine.medical_specialtyGanglion CellsOpen angle glaucomaOcular AnatomyProtein Array AnalysisHemorrhageResearch and Analysis MethodsOptic Disc03 medical and health sciencesSigns and SymptomsDiagnostic MedicineOcular SystemOphthalmologymedicineHumansAgedAutoantibodiesbusiness.industrylcsh:RAutoantibodyBiology and Life SciencesAfferent NeuronsProteinsGlaucomaCell Biologymedicine.diseaseeye diseasesOphthalmology030104 developmental biologyCellular Neuroscience030221 ophthalmology & optometrybiology.proteinlcsh:Qsense organsbusinessNeurosciencePloS one
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