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 首席研究员/项目总监：Tannenbaum, Steven R. 核心 1. 分析、细胞培养和一氧化氮输送。 本计划中项目的一个共同点是定量，成功很大程度上取决于准确的 以及可重复的生物学和生物分析实验。具体需求包括可靠的交付 氧化损伤剂，例如一氧化氮或过氧亚硝酸盐，稳定可靠的细胞培养物，以及 物质的表征和定量分析，包括小极性化合物，例如 甲醛通过核苷、肽和核酸寡聚物转化为完整的蛋白质。不仅是核心1 将这些作为服务提供，确保一致性和连续性，但也直接与 Project 合作 当新方法成为必要或熟悉的方法需要改进时，研究人员就会这样做。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合作开发 核心 1 和项目 1 之间）、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，再次确保实验室之间 一致性，例如在处理细胞系或进行 LC/MS 实验时。核心 1 被全部四个使用 项目（尽管在项目 4 的情况下，这是通过与项目 2 和 3 的交互间接实现的）。项目1 重点关注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 开发和使用输送系统；项目 2 和 3 也使用这些设施，但也使 广泛使用生物分析设施，例如串联四极杆质谱法进行定量 项目 2 和电喷雾飞行时间质谱法用于表征合成和改性 项目 3 中的 DNA 寡聚物。关键人员包括 John S. Wishnok 博士，担任核心和 Laura Trudel 女士负责监督细胞培养和一氧化氮输送设施。