Search results for "methods"

showing 10 items of 4526 documents

MET-EGFR dimerization in lung adenocarcinoma is dependent on EGFR mtations and altered by MET kinase inhibition

2017

Advanced lung cancer has poor survival with few therapies. EGFR tyrosine kinase inhibitors (TKIs) have high response rates in patients with activating EGFR mutations, but acquired resistance is inevitable. Acquisition of the EGFR T790M mutation causes over 50% of resistance; MET amplification is also common. Preclinical data suggest synergy between MET and EGFR inhibitors. We hypothesized that EGFR-MET dimerization determines response to MET inhibition, depending on EGFR mutation status, independently of MET copy number. We tested this hypothesis by generating isogenic cell lines from NCI-H1975 cells, which co-express L858R and T790M EGFR mutations, namely H1975L858R/T790M (EGFR TKI resista…

0301 basic medicineLung NeoplasmsKinase InhibitorsCancer Treatmentlcsh:MedicinePhysical ChemistryBiochemistryFluorophotometryT790MSpectrum Analysis Techniques0302 clinical medicineFluorescence Resonance Energy TransferMedicine and Health SciencesPhosphorylationEnzyme Inhibitorslcsh:ScienceExtracellular Signal-Regulated MAP KinasesEGFR inhibitorsStainingMice Inbred BALB CMultidisciplinaryFluorescent in Situ HybridizationPhysicsCell StainingProto-Oncogene Proteins c-metPrecipitation TechniquesErbB ReceptorsChemistryOncologySpectrophotometry030220 oncology & carcinogenesisPhysical SciencesErlotinibDimerizationProtein BindingResearch Articlemedicine.drugChemical physicsMice NudeMolecular Probe TechniquesAdenocarcinoma of LungAdenocarcinomaBiologyResearch and Analysis Methods03 medical and health sciencesGefitinibGrowth factor receptorCell Line TumormedicineAnimalsHumansImmunoprecipitationMolecular Biology TechniquesLung cancerProtein Kinase InhibitorsMolecular BiologyCell ProliferationCell growthlcsh:RReproducibility of ResultsBiology and Life SciencesDimers (Chemical physics)medicine.diseaseMolecular biologyIsogenic human disease modelsProbe Hybridizationrespiratory tract diseasesHEK293 Cells030104 developmental biologyChemical PropertiesSpecimen Preparation and TreatmentFocal Adhesion Protein-Tyrosine KinasesMutationEnzymologylcsh:QProtein MultimerizationProto-Oncogene Proteins c-aktCytogenetic TechniquesPLOS ONE
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Cellular complexity in MAPK signaling in plants: Questions and emerging tools to answer them

2018

International audience; Mitogen activated protein kinase (MAPK) cascades play an important role in many aspects of plant growth, development, and environmental response. Because of their central role in many important processes, MAPKs have been extensively studied using biochemical and genetic approaches. This work has allowed for the identification of the MAPK genes and proteins involved in a number of different signaling pathways. Less well developed, however, is our understanding of how MAPK cascades and their corresponding signaling pathways are organized at subcellular levels. In this review, we will provide an overview of plant MAPK signaling, including a discussion of what is known a…

0301 basic medicineMAPK/ERK pathwayactivity sensorsPlant growth[SDV]Life Sciences [q-bio]plantComputational biologyPlant ScienceReviewlcsh:Plant culture03 medical and health sciences[SDV.BV]Life Sciences [q-bio]/Vegetal Biologylcsh:SB1-1110biologyphosphorylationsignaling cascade;MAPK;phosphorylation;plant;microscopy;activity sensorsSubcellular localizationMAPKMapk signaling030104 developmental biologyMitogen-activated protein kinasebiology.proteinmicroscopyPhosphorylationSignal transductionExperimental methodssignaling cascade
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Effects of Ranolazine on Astrocytes and Neurons in Primary Culture.

2016

Ranolazine (Rn) is an antianginal agent used for the treatment of chronic angina pectoris when angina is not adequately controlled by other drugs. Rn also acts in the central nervous system and it has been proposed for the treatment of pain and epileptic disorders. Under the hypothesis that ranolazine could act as a neuroprotective drug, we studied its effects on astrocytes and neurons in primary culture. We incubated rat astrocytes and neurons in primary cultures for 24 hours with Rn (10-7, 10-6 and 10-5 M). Cell viability and proliferation were measured using trypan blue exclusion assay, MTT conversion assay and LDH release assay. Apoptosis was determined by Caspase 3 activity assay. The …

0301 basic medicineMacroglial CellsPhysiologyInterleukin-1betaProtein ExpressionCell Culture Techniqueslcsh:MedicineApoptosisPharmacologyPathology and Laboratory Medicine0302 clinical medicineRanolazineAnimal CellsImmune PhysiologyMedicine and Health SciencesEnzyme assaysColorimetric assaysEnzyme-Linked Immunoassayslcsh:ScienceBioassays and physiological analysisImmune ResponseNeuronsInnate Immune SystemMultidisciplinaryMTT assayCell DeathCaspase 3medicine.anatomical_structureCell ProcessesCytokinesTumor necrosis factor alphaCellular TypesAstrocyteResearch ArticleProgrammed cell deathCell SurvivalImmunologyCaspase 3Glial CellsBiologyGene Expression Regulation EnzymologicMitochondrial Proteins03 medical and health sciencesSigns and SymptomsmedicineGene Expression and Vector TechniquesAnimalsMTT assayViability assayMolecular Biology TechniquesImmunoassaysMolecular BiologyInflammationMolecular Biology Assays and Analysis TechniquesSuperoxide DismutaseTumor Necrosis Factor-alphalcsh:RBiology and Life SciencesCell BiologyMolecular DevelopmentRatsPPAR gammaResearch and analysis methodsOxidative Stress030104 developmental biologyCell cultureApoptosisAstrocytesImmune SystemBiochemical analysisImmunologic Techniqueslcsh:QFisiologia humanaApoptosis Regulatory ProteinsCarrier Proteins030217 neurology & neurosurgeryDevelopmental BiologyPloS one
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Substantial deficiency of free sialic acid in muscles of patients with GNE myopathy and in a mouse model

2017

GNE myopathy (GNEM), also known as hereditary inclusion body myopathy (HIBM), is a late- onset, progressive myopathy caused by mutations in the GNE gene encoding the enzyme responsible for the first regulated step in the biosynthesis of sialic acid (SA). The disease is characterized by distal muscle weakness in both the lower and upper extremities, with the quadriceps muscle relatively spared until the late stages of disease. To explore the role of SA synthesis in the disease, we conducted a comprehensive and systematic analysis of both free and total SA levels in a large cohort of GNEM patients and a mouse model. A sensitive LC/MS/MS assay was developed to quantify SA in serum and muscle h…

0301 basic medicineMaleBiopsylcsh:MedicineMuscle ProteinsBiochemistryPathogenesischemistry.chemical_compoundMice0302 clinical medicineTandem Mass SpectrometryMedicine and Health Scienceslcsh:ScienceMusculoskeletal SystemMultidisciplinarymedicine.diagnostic_testOrganic CompoundsMusclesGastrocnemius MusclesAnimal ModelsMuscle AnalysisMiddle AgedChemistrymedicine.anatomical_structureBioassays and Physiological AnalysisBiochemistryExperimental Organism SystemsPhysical SciencesFemalemedicine.symptomAnatomyResearch ArticleMuscle tissueAdultmedicine.medical_specialtyAdolescentMuscle TissueMouse ModelsSurgical and Invasive Medical ProceduresCreatineResearch and Analysis Methods03 medical and health sciencesYoung AdultModel OrganismsInternal medicineBiopsymedicineAnimalsHumansMyopathyMuscle SkeletalAgedHereditary inclusion body myopathybusiness.industrylcsh:ROrganic ChemistryChemical CompoundsBiology and Life SciencesProteinsmedicine.diseaseCreatineN-Acetylneuraminic AcidSialic acidDistal MyopathiesDisease Models Animal030104 developmental biologyEndocrinologyBiological TissuechemistrySkeletal Muscleslcsh:QbusinessN-Acetylneuraminic acid030217 neurology & neurosurgeryBiomarkersChromatography LiquidPLoS ONE
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The maternal hormone in the male brain: Sexually dimorphic distribution of prolactin signalling in the mouse brain.

2018

Research of the central actions of prolactin is highly focused on females, but this hormone has also documented roles in male physiology and behaviour. Here, we provide the first description of the pattern of prolactin-derived signalling in the male mouse brain, employing the immunostaining of phosphorylated signal transducer and activator of transcription 5 (pSTAT5) after exogenous prolactin administration. Next, we explore possible sexually dimorphic differences by comparing pSTAT5 immunoreactivity in prolactin-supplemented males and females. We also assess the role of testosterone in the regulation of central prolactin signalling in males by comparing intact with castrated prolactin-supp…

0301 basic medicineMaleCell signalingPeptide HormonesSignal transductionBiochemistrychemistry.chemical_compoundMice0302 clinical medicineArcuate NucleusSTAT5 Transcription FactorMedicine and Health SciencesMorphogenesisTestosteroneLipid HormonesPhosphorylationTestosteroneNeuronsSex CharacteristicsMultidisciplinarySexual DifferentiationCerebrumReproductionQRBrainHormones esteroidesSTAT signalingmedicine.anatomical_structureCervell Localització de funcionsHypothalamusAndrogensMedicineFemaleAnatomyhormones hormone substitutes and hormone antagonistsResearch Articlemedicine.medical_specialtyendocrine systemCell biologyScienceHypothalamusBiologyResearch and Analysis MethodsAmygdala03 medical and health sciencesInternal medicinemedicineAnimalsCastrationImmunohistochemistry TechniquesSexual DimorphismProlactin receptorBiology and Life SciencesProlactinHormonesProlactinSexual dimorphismHistochemistry and Cytochemistry Techniques030104 developmental biologyEndocrinologyCastrationchemistryImmunologic Techniques030217 neurology & neurosurgeryHormoneDevelopmental BiologyPloS one
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Performance of a rapid diagnostic test for the detection of Cryptosporidium spp. in African children admitted to hospital with diarrhea

2020

Background Cryptosporidium is a protozoan parasite that causes mild to severe diarrhoeal disease in humans. To date, several commercial companies have developed rapid immunoassays for the detection of Cryptosporidium infection. However, the challenge is to identify an accurate, simple and rapid diagnostic tool for the estimation of cryptosporidiosis burden. This study aims at evaluating the accuracy of CerTest Crypto, a commercialized rapid diagnostic test (RDT) for the detection of Cryptosporidium antigens in the stool of children presenting with diarrhoea. Methods A cross-sectional study was conducted in four study sites in Sub-Saharan Africa (Gabon, Ghana, Madagascar, and Tanzania), from…

0301 basic medicineMaleCryptosporidium infectionCross-sectional studyRC955-962CryptosporidiosisArtificial Gene Amplification and ExtensionPathology and Laboratory MedicinePolymerase Chain ReactionENTAMOEBA-HISTOLYTICAGeographical LocationsFeces0302 clinical medicineArctic medicine. Tropical medicineMedicine and Health SciencesMedicineProtozoansRapid diagnostic testbiologyEukaryotaCryptosporidiumDiarrheaInfectious DiseasesCryptosporidium parvumChild PreschoolGIARDIA-LAMBLIAFemalemedicine.symptomPublic aspects of medicineRA1-1270BURDENPolymorphism Restriction Fragment LengthResearch ArticleDiarrheamedicine.medical_specialty030231 tropical medicineANTIGENCryptosporidiumGastroenterology and HepatologyResearch and Analysis MethodsSensitivity and Specificity03 medical and health sciencesSigns and SymptomsDiagnostic MedicineOOCYSTSInternal medicineparasitic diseasesParasitic DiseasesHumansGabonMolecular Biology TechniquesMolecular BiologyAfrica South of the Saharabusiness.industryPublic Health Environmental and Occupational HealthOrganismsCryptosporidium ParvumBiology and Life SciencesInfantGold standard (test)biology.organism_classificationmedicine.diseaseConfidence intervalParasitic Protozoans030104 developmental biologyCross-Sectional StudiesFECAL SAMPLESPeople and PlacesAfricabusiness
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Expression quantitative trait loci for PAX8 contributes to the prognosis of hepatocellular carcinoma

2017

Paired-box family member PAX8 encodes a transcription factor that has a role in cell differentiation and cell growth and may participate in the prognosis of hepatocellular carcinoma (HCC). By bioinformatics analysis, we identified several single nucleotide polymorphisms (SNPs) within a newly identified long non-coding RNA (lncRNA) AC016683.6 as expression quantitative trait loci (eQTLs) for PAX8. Hence, we hypothesized that PAX8eQTLs in lncRNA AC016683.6 may influence the HCC prognosis. We then performed a case-only study to assess the association between the two SNPs as well as the prognosis of HCC in 331 HBV-positive HCC patients without surgical treatment. Cox proportional hazard models …

0301 basic medicineMaleHeredityPaired BoxCancer Treatmentlcsh:MedicineBiochemistry0302 clinical medicineMathematical and Statistical TechniquesMedicine and Health Scienceslcsh:ScienceMultidisciplinaryPharmaceuticsLiver DiseasesLiver NeoplasmsMiddle AgedPrognosisNucleic acidsSurvival RateGenetic MappingOncology030220 oncology & carcinogenesisHepatocellular carcinomaPhysical SciencesRegression AnalysisFemaleLiver cancerStatistics (Mathematics)Research ArticleCarcinoma HepatocellularGenotypeQuantitative Trait LociSingle-nucleotide polymorphismVariant GenotypesGastroenterology and HepatologyResearch and Analysis MethodsCarcinomasPolymorphism Single Nucleotide03 medical and health sciencesPAX8 Transcription FactorProtein DomainsDrug TherapyDiagnostic MedicineGastrointestinal TumorsCarcinomamedicineGeneticsChemotherapyHumansGenetic Predisposition to DiseaseAlleleStatistical MethodsNon-coding RNASurvival rateSurvival analysisAllelesbusiness.industrylcsh:RCancers and NeoplasmsBiology and Life SciencesProteinsHepatocellular Carcinomamedicine.disease030104 developmental biologyExpression quantitative trait lociCancer researchLong non-coding RNAsRNAlcsh:QbusinessMathematicsPLoS ONE
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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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