Search results for "blotting"

showing 10 items of 899 documents

Corynebacterium parvum (Propionibacterium acnes): an inducer of tumor necrosis factor-alpha in human peripheral blood mononuclear cells and monocytes…

1990

The present study investigates the potential capacity of the immunostimulant Corynebacterium parvum (C.p.) to induce tumor necrosis factor-alpha (TNF-alpha) in human peripheral blood mononuclear cells (PBMC) and blood monocytes (BMo) in vitro. Both at the mRNA and protein level, stimulation of PBMC and BMo upon C.p. induces TNF-alpha. Compared to the hitherto used TNF-alpha inducers in vitro such as Sendai virus, phytohemagglutinin or lipopolysaccharide the C.p. stimulus displayed a threefold stronger induction of TNF-alpha production (p less than 0.001). Using C.p. as an inducer it was possible to demonstrate that TNF-alpha production is regulated by prostaglandin E2; preincubation of the …

Lipopolysaccharidesmedicine.drug_classLymphocyteImmunologyEnzyme-Linked Immunosorbent AssayBiologyIn Vitro TechniquesPeripheral blood mononuclear cellImmunostimulantDinoprostoneMonocytesInterferon-gammamedicineImmunology and AllergyHumansInterferon gammaInducerPropionibacterium acnesProstaglandin E2Cells CulturedDose-Response Relationship DrugTumor Necrosis Factor-alphaMonocyteBlotting NorthernMolecular biologymedicine.anatomical_structureImmunologyLeukocytes MononuclearRNATumor necrosis factor alphaImmunizationDNA Probesmedicine.drugEuropean journal of immunology
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Protection by nitric oxide against liver inflammatory injury in animals carrying a nitric oxide synthase-2 transgene

2001

22 pages, 7 figures, 1 table.

Lipopolysaccharidesmedicine.medical_specialtyLipopolysaccharideTransgeneBlotting WesternNitric Oxide Synthase Type IIApoptosisGalactosamineMice TransgenicLipopolysaccharideNitric OxideBiochemistryLiver cellsProinflammatory cytokineNitric oxidechemistry.chemical_compoundMiceInternal medicineGeneticsmedicineAnimalsTransgenesPromoter Regions GeneticMolecular BiologyLiver injurybiologyTumor Necrosis Factor-alphaNF-kappa BNitric oxide synthase 2medicine.diseaseEndotoxinsEndocrinologychemistryLiverbiology.proteinTumor necrosis factor alphaNitric Oxide SynthasePhosphoenolpyruvate carboxykinaseFood DeprivationBiotechnologyInterleukin-1
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A human renal cancer line as a new antigen source for the detection of antibodies to cytoplasmic and nuclear antigens in sera of patients with Wegene…

1991

Autoantibodies directed against cytoplasmic antigens of neutrophils (ANCA), especially proteinase 3 (C-ANCA), have proved to be a useful clinical tool to support the diagnosis or to monitor disease activity in Wegener's granulomatosis (WG). Till now, human neutrophil granulocytes have represented the major antigen source used to detect antibodies in WG by the immunofluorescence technique (IFT). We have tested serum samples of 164 patients with different connective tissue diseases (50 suffering from clinically active WG) performing IFT on a human renal cancer line (SK-RC11) and have found antibodies against the nuclear and cytoplasmic antigens in 39 patients. C-ANCA+ sera displayed a charact…

Liver CirrhosisTime Factorsmedicine.drug_classNeutrophilsImmunologyBlotting WesternFluorescent Antibody TechniqueImmunofluorescenceMonoclonal antibodyAutoantigensMonocytesSerologyCell LineArthritis RheumatoidScleroderma LocalizedAntigenProteinase 3medicineImmunology and AllergyHumansLupus Erythematosus SystemicAnti-neutrophil cytoplasmic antibodyAutoantibodiesMixed Connective Tissue Diseasemedicine.diagnostic_testbiologyTumor Necrosis Factor-alphaGranulomatosis with PolyangiitisBiological Transportmedicine.diseaseVirologyMolecular biologyKidney NeoplasmsSjogren's SyndromeAntibodies Antinuclearbiology.proteinInterferonsAntibodyGranulomatosis with polyangiitisGranulocytesJournal of immunological methods
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Antigen-presenting function and B7 expression of murine sinusoidal endothelial cells and Kupffer cells.

1996

Abstract BACKGROUND & AIMS: Inflammatory liver disease as well as rejection of liver allografts are thought to be mediated by resident antigen- presenting cells in the liver. At the same time, in vivo antigen presentation in the liver appears to be a more tolerogenic than systemic antigen challenge. The aim of this study was to show and characterize the antigen-presenting capability of sinusoidal endothelial cells and Kupffer cells. METHODS: Purified murine sinusoidal endothelial cells and Kupffer cells were studied for their ability to serve as accessory cells and antigen-presenting cells by proliferation assays. They were also studied for their expression of interleukin 1 and the B7 costi…

Liver cytologyKupffer CellsAntigen presentationMolecular Sequence DataAntigen-Presenting CellsBiologyLymphocyte ActivationPolymerase Chain ReactionMicemedicineAnimalsRNA MessengerAntigen-presenting cellInterleukin 3Antigen PresentationMice Inbred BALB CCD40HepatologyBase SequenceKupffer cellGastroenterologyBlotting NorthernCell biologyInterleukin-10RatsInterleukin 33medicine.anatomical_structureLiverImmunologybiology.proteinInterleukin 12B7-1 AntigenEndothelium VascularInterleukin-1Gastroenterology
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Centrifugation does not alter spatial distribution of `BEP4' mRNA in paracentrotus lividus EGG

1997

AbstractParacentrotus lividus unfertilized eggs were centrifuged in a sucrose gradient, so to split each into two parts: a nucleated light fragment and an anucleated heavy fragment. Northern blot analyses utilizing a bep4 probe as animal marker and H2A histone gene and 12S-mit RNA as controls indicate that the eggs are elongated along the animal-vegetal axis during centrifugation and thereafter split into an animal and a vegetal half. Treatment of the eggs with colchicine before centrifugation abolishes the animal localization of bep4 mRNA.

Localized mRNABiophysicsCentrifugationCell FractionationBiochemistryMicrotubulesParacentrotus lividuschemistry.chemical_compoundStructural BiologyBotanyA/V axisGeneticsCentrifugation Density GradientColchicineAnimalsCentrifugationNorthern blotRNA MessengerMolecular BiologyGeneIn Situ HybridizationOvumMessenger RNAbiologyUnfertilized EggsRNACell PolarityMembrane ProteinsCell Biologybiology.organism_classificationBlotting NorthernMolecular biologychemistrySea Urchinsembryonic structuresCortexColchicineFEBS Letters
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Upon oxidative stress, the antiapoptotic Hsp60/procaspase-3 complex persists in mucoepidermoid carcinoma cells.

2008

Hsp60, a mitochondrial chaperonin highly conserved during evolution, has been found elevated in the cytosol of cancer cells, both in vivo and in vitro, but its role in determining apoptosis during oxidative stress (OS) has not yet been fully elucidated. The aim of the present work was to study the effects of OS on Hsp60 levels and its interactions with procaspase- 3 (p-C3) and p53 in tumor cells. NCI-H292 (mucoepidermoid carcinoma) cells were exposed to various concentrations of hydrogen peroxide (H2O2) for 24 hours. Cell viability was determined by Trypan blue and MTT assays. DNA damage was assessed by the Comet assay, and apoptosis was measured by the AnnexinV cytofluorimetric test. Expos…

Lung Neoplasmsanimal structuresHistologyCell SurvivalDNA damageBlotting WesternBiophysicsHsp60;procaspase-3;mucoepidermoid carcinomaGene ExpressionTetrazolium SaltsApoptosisBiologymedicine.disease_causechemistry.chemical_compoundCell Line TumormedicineHumansChaperonin Hsp60 Cpn60 procaspase-3 caspase- 3 DNA damage p53 apoptosis.Viability assaylcsh:QH301-705.5FormazansCaspase 3Settore BIO/16 - Anatomia UmanaChaperonin 60DNAHydrogen PeroxideTrypan BlueCell BiologyImmunohistochemistryMolecular biologyComet assayOxidative Stresslcsh:Biology (General)chemistryApoptosisCancer cellCarcinoma MucoepidermoidHSP60Trypan blueComet AssayTumor Suppressor Protein p53Oxidative stressDNA Damage
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RGD motifs on the surface of baculovirus enhance transduction of human lung carcinoma cells.

2006

Baculovirus vectors have been shown to enter a variety of mammalian cell lines and gene transfer with wild-type baculovirus (WT) has been demonstrated both in vitro and in vivo. Different protein motifs have been displayed on the viral surface to serve as ligands for cell-specific receptor molecules. We have generated recombinant baculovirus vectors displaying an RGD-motif, recognized by alphaV integrin, on the viral surface. The RGD motifs within the C-terminus of coxsackie virus A9 and human parechovirus 1 VP1 proteins were fused to the N-terminus of the major envelope glycoprotein, gp64, of Autographa californica multiple nucleopolyhedrovirus. The recombinant RGD-presenting viruses bound…

Lung NeoplasmsvirusesRecombinant Fusion ProteinsIntegrinBlotting WesternGenetic VectorsBioengineeringPlasma protein bindingTransfectionApplied Microbiology and Biotechnologylaw.inventionTransduction (genetics)lawCell Line TumorAnimalsHumansCells CulturedRGD motifMicroscopy ConfocalbiologyModels GeneticGeneral MedicineTransfectionMolecular biologyIntegrin alphaVbiology.proteinRecombinant DNALight emissionElectrophoresis Polyacrylamide GelBaculoviridaeOligopeptidesBiotechnologyProtein BindingJournal of biotechnology
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The synthesis of SNAT2 transporters is required for the hypertonic stimulation of system A transport activity

2004

AbstractIn cultured human fibroblasts incubated under hypertonic conditions, the stimulation of system A for neutral amino acid transport, associated to the increased expression of the mRNA for SNAT2 transporter, leads to an expanded intracellular amino acid pool and to the recovery of cell volume. A protein of nearly 60 kDa, recognized by an antiserum against SNAT2, is increased both in the pool of biotinylated membrane proteins and in the total cell lysate of hypertonically stressed cells. The increased level of SNAT2 transporters in hypertonically stressed cells is confirmed by immunocytochemistry. DRB, an inhibitor of transcription, substantially inhibits the increase of SNAT2 proteins …

LysisAmino Acid Transport System AProlineTranscription GeneticGlutamineBlotting WesternHypertonic SolutionsBiophysicsStimulationBiologyHuman fibroblastBiochemistrySubstrate SpecificityAmino acid starvationHypertonic stressCell volumeNeutral amino acid transportHumansBiotinylationRNA MessengerCells CulturedCell Sizechemistry.chemical_classificationRadioisotopesCell MembraneBiological TransportPhosphorusCell BiologyFibroblastsImmunohistochemistryAmino acidGlutamineMolecular WeightKineticschemistryBiochemistryMembrane proteinHypertonic StressIntracellularDichlororibofuranosylbenzimidazoleBiochimica et Biophysica Acta (BBA) - Biomembranes
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Optimized RNA extraction and northern hybridization in streptomycetes.

2010

Abstract Northern blot hybridization is a useful tool for analyzing transcript patterns. To get a picture of what really occurs in vivo, it is necessary to use a protocol allowing full protection of the RNA integrity and recovery and unbiased transfer of the entire transcripts population. Many protocols suffer from severe limitations including only partial protection of the RNA integrity and/or loss of small sized molecules. Moreover, some of them do not allow an efficient and even transfer in the entire sizes range. These difficulties become more prominent in streptomycetes, where an initial quick lysis step is difficult to obtain. We present here an optimized northern hybridization protoc…

LysisPopulationtotal RNA purificationComputational biologyBiologynorthern hybridizationSettore BIO/19 - Microbiologia GeneraleStreptomycesGeneral Biochemistry Genetics and Molecular BiologyRNA degradationNorthern bloteducationlcsh:QH301-705.5GeneticsRNA glyoxylationeducation.field_of_studylcsh:R5-920streptomyceteBiochemistry Genetics and Molecular Biology(all)alkaline blottingMethodologyRNAbiology.organism_classificationBlotNorthern hybridizationSettore BIO/18 - Geneticalcsh:Biology (General)RNA processingstreptomycetesRNA extractionlcsh:Medicine (General)Biological procedures online
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Cajaninstilbene acid (CSA) exerts cytoprotective effects against oxidative stress through the Nrf2-dependent antioxidant pathway.

2013

Cajaninstilbene acid (CSA), an active compound separated from pigeon pea leaves, possesses the highly efficient antioxidant activities. Transcription factor nuclear factor-erythroid 2-related factor 2 (Nrf2) is an important regulator of cellular oxidative stress. This study examined the role of Nrf2 in CSA-mediated antioxidant effects on human hepatocarcinoma (HepG2) cell line. The generation of reactive oxygen species (ROS) upon H2O2 and CSA treatment was lower than that of H2O2 alone. CSA activated Nrf2 as evaluated by Western blotting. A luciferase reporter assay also demonstrated that CSA-activated signaling resulted in the increased transcriptional activity of Nrf2 through binding to t…

MAPK/ERK pathwayAntioxidantNF-E2-Related Factor 2medicine.medical_treatmentBlotting WesternBiologyToxicologymedicine.disease_causeenvironment and public healthAntioxidantsStilbenesmedicineNAD(P)H Dehydrogenase (Quinone)HumansProtein kinase BTranscription factorPI3K/AKT/mTOR pathwaychemistry.chemical_classificationReactive oxygen speciesGeneral MedicineHep G2 Cellsrespiratory systemAntioxidant Response ElementsSalicylatesOxidative StressBiochemistrychemistryCytoprotectionNAD+ kinaseOxidative stressHeme Oxygenase-1Signal TransductionToxicology letters
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