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.
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