Core: Analytical, Cell Culture, Nitric Oxide Delivery

核心：分析、细胞培养、一氧化氮输送

• 批准号: 7514464
• 负责人: JOHN S. WISHNOK
• 金额: $ 28.96万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7514464
• 关键词: 3-nitrotyrosine; Albumins; Ammonium; Animal Model; Antibodies; Area; Arts; Atmospheric Pressure; Behavior; Biochemistry; Biological; Biological Markers; Biological Models; Buffers; Cell Culture Techniques; Cell Line; Chemicals; Chromatography; Clinical; Cluster Analysis; Collaborations; Collection; Complex Mixtures; Computer software; DNA; Data; Data Analyses; Deoxyribose; Detection; Development; Digestion; Disease; Dithiothreitol; Ethanol; Evaluation; Exposure to; Formaldehyde; Formalin; Freezing; Funding; Gene Expression; Goals; Hand; Heating; High Pressure Liquid Chromatography; Histocompatibility Testing; Human; Human Resources; Hydrolysis; Ice; Imagery; Immersion Investigative Technique; Immunohistochemistry; In Vitro; Incubated; Individual; Inflammation; Ions; Laboratories; Lead; Licensing; Liquid substance; Mass Spectrum Analysis; Measures; Methods; Microscope; Microtomy; Molecular Profiling; Nitrates; Nitric Oxide; Nitrites; Nucleic Acids; Nucleosides; Organic solvent product; Outcome; Paraffin; Paraffin Embedding; Pathology; Peptide Nucleic Acids; Peptides; Peroxonitrite; Physiological; Potassium; Preparation; Principal Investigator; Procedures; Process; Proteins; Reaction; Recovery; Relative (related person); Reliance; Reporting; Research Personnel; Resolution; Resources; Running; Sampling; Scanning; Sensitivity and Specificity; Series; Serum Albumin; Services; Signal Transduction; Slide; Sodium; Solutions; Source; Supervision; System; Techniques; Technology; Time; Tissue Sample; Tissues; Training; Trypsin; Tyrosine; Water; Work; Xylene; adduct; analytical method; animal tissue; aqueous; base; carcinogenesis; design; dityrosine; improved; indexing; insight; instrument; interest; ion source; ionization; liquid chromatography mass spectrometry; man; mass spectrometer; method development; microbial alkaline proteinase inhibitor; multiple reaction monitoring; nitration; numb protein; oxidation; oxidative damage; programs; protein aminoacid sequence; protein expression; research study; sample fixation; software development; spiroiminodihydantoin; statistics; success; tissue fixing; urinary

Continuation Page Principal Investigator/ProgramDirector: Tannenbaum, Steven R. Core 1. Analytical, Cell Culture, and Nitric Oxide Delivery. 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 1and 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.

续页首席调查员/方案主任：Tannenbaum，Steven R. 核心1.分析、细胞培养和一氧化氮输送。 这个项目的一个共同主线是量化，而成功在很大程度上取决于准确性 以及可重复的生物和生物分析实验。具体需求包括可靠的交付 氧化损害剂，如一氧化氮或过氧亚硝酸盐，稳定和可靠的细胞培养，以及两者 表征和定量分析各种物质，从小的、极性的化合物，如 甲醛通过核苷、多肽和核酸低聚物，到达完整的蛋白质。核心1不仅 将这些作为服务提供，确保一致性和连续性，但也直接与Project协作 研究人员作为新的方法变得必要或熟悉的方法需要改进。设施包括 独特的一氧化氮传递系统(在以前的供资期间作为合作开发 在核心1和项目1之间)，集中式细胞培养设施和最先进的层析和 质谱学实验室。核心还提供培训和监督，再次确保实验室到实验室 一致性，例如在处理细胞系时，或在进行LC/MS实验时。核心1被所有四个人使用 项目(尽管这是通过与项目2和项目3的交互间接进行的，在项目4的情况下)。项目1 专注于与炎症相关的活性物种的行为，并使用细胞培养设施和两者 开发和使用交付系统；项目2和3也使用这些设施，但也会 广泛使用生物分析设施，例如串联四极杆质谱仪，用于对 项目2和电喷雾飞行时间质谱仪用于表征合成和改性 项目3中的DNA寡聚体。主要人员包括约翰·S·威什诺克博士担任核心和 负责细胞培养和一氧化氮输送设施的劳拉·特鲁德尔女士。