Core: Analytical, Cell Culture, Nitric Oxide Delivery

核心：分析、细胞培养、一氧化氮输送

基本信息

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
项目类别：
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

项目摘要

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。