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Core: Analytical, Cell Culture, Nitric Oxide Delivery

Core: Analytical, Cell Culture, Nitric Oxide Delivery
核心:分析、细胞培养、一氧化氮输送
批准号:
8555149
负责人:
JOHN S. WISHNOK
金额:
$28.7万
依托单位国家:
美国
项目类别:
财政年份:
1997
资助国家:
美国
项目状态:
已结题
起止时间:
1997-01-15 至 2013-12-31
关键词:
3-nitrotyrosineAlbuminsAmmoniumAnimal ModelAntibodiesAreaAtmospheric PressureBehaviorBiochemistryBiologicalBiological MarkersBiological ModelsBuffersCell Culture TechniquesCell LineChemicalsChromatographyClinicalCluster AnalysisCollaborationsCollectionComplex MixturesComputer softwareDNADataData AnalysesDeoxyriboseDetectionDevelopmentDigestionDiseaseDithiothreitolEthanolEvaluationExposure toFormaldehydeFormalinFreezingFundingGene ExpressionGoalsHandHeatingHigh Pressure Liquid ChromatographyHistocompatibility TestingHumanHuman ResourcesHydrolysisIceImageryImmersion Investigative TechniqueImmunohistochemistryIn VitroIncubatedIndividualInflammationIonsLaboratoriesLeadLicensingLiquid substanceMass Spectrum AnalysisMeasuresMethodsMicroscopeMicrotomyMolecular ProfilingNitratesNitric OxideNitritesNucleic AcidsNucleosidesOrganic solvent productOutcomeParaffinParaffin EmbeddingPathologyPeptide Nucleic AcidsPeptidesPeroxonitritePhysiologicalPotassiumPreparationProceduresProcessProteinsReactionRecoveryRelative (related person)RelianceReportingResearch PersonnelResolutionResourcesRunningSamplingScanningSensitivity and SpecificitySeriesSerum AlbuminServicesSignal TransductionSlideSodiumSolutionsSourceSupervisionSystemTechniquesTechnologyTimeTissue SampleTissuesTrainingTrypsinTyrosineWaterWorkXyleneadductanalytical methodanimal tissueaqueousbasecarcinogenesisdesigndityrosineimprovedindexinginsightinstrumentinterestion sourceionizationliquid chromatography mass spectrometrymanmass spectrometermethod developmentmicrobial alkaline proteinase inhibitormultiple reaction monitoringnitrationoxidationoxidative damageprogramsprotein aminoacid sequenceprotein expressionresearch studysample fixationsoftware developmentspiroiminodihydantoinstatisticssuccesstissue fixingurinary

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中文摘要
翻译
这个项目的一个共同点是定量,成功在很大程度上取决于准确和可重复的生物和生物分析实验。具体需求包括氧化损伤剂(如一氧化氮或过氧亚硝酸盐)的可靠输送,稳定可靠的细胞培养,以及从小的极性化合物(如甲醛)到核苷、多肽和核酸低聚物,再到完整蛋白质的表征和定量分析。核心1不仅提供这些服务,确保一致性和连续性,而且当新方法成为必要或熟悉的方法需要改进时,还直接与项目研究人员合作。设施包括独特的一氧化氮输送系统(在之前的资助期间作为核心1和项目1之间的合作开发),集中式细胞培养设施以及最先进的色谱和质谱实验室。核心还提供培训和监督,再次确保实验室到实验室的一致性,例如在处理细胞系或进行LC/MS实验时。核心1由所有四个项目使用(尽管在项目4的情况下,这是通过与项目2和3的交互间接使用的)。项目1侧重于炎症相关活性物种的行为,使用细胞培养设施,开发和使用传递系统;项目2和项目3也使用这些设备,但也广泛使用生物分析设备,例如,项目2中用于定量的串联四极杆质谱和项目3中用于表征合成和修饰DNA低聚物的电喷雾飞行时间质谱。主要人员包括John S. Wishnok博士作为核心的总主管,Laura Trudel女士负责监督细胞培养和一氧化氮输送设施。
英文摘要
A thread common to the projects in this Program is quantitation, and success depends strongly on accurate and reproducible biological and bioanalytical experiments. The specific needs include dependable delivery of oxidative damage agents, e.g., nitric oxide or peroxynitrite, stable and reliable cell cultures, and both characterization and quantitative analysis of substances ranging from small, polar compounds such as formaldehyde through nucleosides, peptides, and nucleic-acid oligomers, to intact proteins. Core 1 not only provides these as services, assuring both consistency and continuity, but also works directly with Project researchers as new methods become necessary or familiar methods require improvement. Facilities include unique nitric oxide delivery systems (that were developed during previous funding periods as a collaboration between Core 1 and Project 1), centralized cell-culture facilities, and state-of-the-art chromatography and mass spectrometry laboratories. The Core also provides training and supervision, again assuring lab-to-lab consistency, e.g., in handling cell-lines, or in conducting LC/MS experiments. Core 1 is used by all four Projects (although this is indirect via interaction with Projects 2 and 3 in the case of Project 4). Project 1 focuses on the behavior of inflammation-related active species and uses the cell culture facilities and both develops and uses the delivery systems; Projects 2 and 3 use these facilities as well, but also make extensive use of the bioanalytical facilities, e.g., tandem quadrupole mass spectrometry for quantitation in Project 2 and electrospray time-of-flight mass spectrometry for characterization of synthetic and modified DNA oligomers in Project 3. Key personnel include Dr. John S. Wishnok as overall director of the Core and Ms. Laura Trudel who oversees the cell culture and nitric oxide delivery facilities.
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Core: Analytical, Cell Culture, Nitric Oxide Delivery
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