To understand how neoplastic cells adapt to stresses commonly encountered in solid and hematopoietic malignancies, it is critical to establish how cells alter their metabolism in order to survive these stresses. The purpose of the Metabolism Core is to provide support to the Project Leaders for analyses of cellular metabolism under specific stresses that reflect the unique features of tumor environments: specifically, growth factor withdrawal, nutrient and/or oxygen depletion. The core will provide advice, training, and access to equipment and reagents essential to these inquiries. The core will provide assistance with assays for glucose uptake, glycolysis, NADH and NADPH production, oxygen consumption, and mitochondrial function. Many of the proposed experiments also depend on the use of customized work stations and CO2 incubators for cell culture under hypoxic and/ or anoxic conditions. Furthermore, sucrose gradient centrifugation for polysome profiling will be supervised by a research technician within the core. Therefore, the core will maintain and support the following equipment: A. oxymeter, B. fluorometer, C. hypoxia hoods and specialized hypoxia C02 incubators, D. gradient makers and rotors for sucrose gradient centrifugation. Funds for a second oxymeter, and an anoxia workstation capable of achieving O2 levels_<0.01 % O2, are also requested. The core director, Dr. Brian Keith, will supervise a research specialist in these endeavors. Both core personnel have multiple publications in the areas of cellular bioenergetics, hypoxia and metabolism. The goal of this core is to provide equipment shared by all three research programs and train research associates, postdoctoral fellows, graduate students, and technicians in the techniques described.
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