Core: Analytical, Cell Culture, Nitric Oxide Delivery

该计划项目的一个共同主线是定量，而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。具体需求包括可靠地提供氧化损害剂，例如一氧化氮或过氧亚硝酸盐，稳定和可靠的细胞培养，以及从小的极性化合物，如甲醛，到核苷、多肽和核酸低聚物，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，而且当新方法变得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(在之前的供资期间作为核心1和项目1之间的合作而开发)、中央细胞培养设施以及最先进的层析和质谱仪实验室。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，在处理细胞系方面，或在进行LC/MS实验时。所有四个项目都使用核心1(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重点研究与炎症相关的活性物种的行为，使用细胞培养设施并开发和使用输送系统；项目2和项目3也使用这些设施，但也广泛使用生物分析设施，例如项目2中用于定量的串联四极杆质谱仪和项目3中用于表征合成和修饰的DNA低聚物的电喷雾飞行时间质谱仪。主要人员包括核心中心的总主任John S.Wishnok博士和负责细胞培养和一氧化氮输送设施的Laura Trudel女士。

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.