Core: Analytical, Cell Culture, Nitric Oxide Delivery

该计划中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的可靠递送，例如，一氧化氮或过氧亚硝酸盐，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、肽和核酸低聚物到完整蛋白质的物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，而且还直接与项目研究人员合作，因为新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输送系统（在以前的资助期间作为核心1和项目1之间的合作开发），集中细胞培养设施，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的一致性，例如，在处理细胞系或进行LC/MS实验中。所有四个项目都使用核心1（尽管在项目4中，这是通过与项目2和项目3的互动间接使用的）。项目1侧重于炎症相关活性物质的行为，并使用细胞培养设施，开发和使用递送系统;项目2和3也使用这些设施，但也广泛使用生物分析设施，例如，在项目2中，串联四极杆质谱用于定量，在项目3中，电喷雾飞行时间质谱用于表征合成和修饰的DNA寡聚体。主要人员包括约翰·S。Wishnok担任核心的总经理，Laura Trudel女士负责监督细胞培养和一氧化氮输送设施。

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.