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INVESTIGATOR: Farnell, Y. Z.

PERFORMING INSTITUTION:
MISSISSIPPI STATE UNIV
MISSISSIPPI STATE, MISSISSIPPI 39762

THE ULTRACENTRIFUGE SYSTEM: A CRITICAL TOOL TO ENHANCE THE PERFORMANCE OF AGRICULTURAL RELATED RESEARCH

NON-TECHNICAL SUMMARY: Researchers in agriculture-related fields often must work not only with plant and animal cells, but also with the biological structures and chemical components within those cells. Ultracentrifugation is a fundamental and widely used tool to separate these tiny structures both from the cells themselves and from each other according a property called differential density. This way, as the centrifuge spins a liquid, it will force different structures with different density to either stay in the liquid or sediment at the bottom of the tube into what's known as a pellet. Ultracentrifugation provides a simple but versatile method to isolate structures of interest, like extracellular vesicles (exosomes), viruses, subcellular organelles, and large pieces of DNA, making it an essential platform to isolate specific molecules as well as quantify them. This proposal seeks funds to help support replacement of an aged and mechanically unsound ultracentrifuge used by a diverse group of researchers. Projects that will benefit from the replacement of the ultracentrifuge range from studies of human metabolic disorders to cattle and poultry diseases to the creation of drought-resistant crops. Obtaining a state-of-the-art ultracentrifuge system (in this case, the proposed Beckman Optima XPN ultracentrifuge) will have an immediate and profound impact on current and planned research into a variety of important agricultural problems. The device will improve routine analyses of small RNAs and proteins associated with exosomes, molecular markers reflective of plant and animal health, and pathogens of livestock and crops. Replacement of the current ultracentrifuge, which is increasingly unreliable and unserviceable, will also reduce a significant bottleneck that limits the rate of progress on several important projects. Lastly, it will allow these research teams to explore new approaches and projects that require a high-quality method of ultracentrifugation.

OBJECTIVES: Project #1. Identification of the exosomal biomarkers for cardiovascular disease in a porcine model with the metabolic syndrome.1) to purify exosomes from conditioned media of primary cardiac fibroblasts from both male MetS/diabetic pigs and nondiabetic control littermates; 2) to examine their characteristics by transmission electron microscope and Western Blot analysis; and 3) to compare the differences of exosomal protein profiles from cardiac fibroblasts of diabetic and nondiabetic pigs by mass spectrometric analysis.Project #2. Development of Molecular Markers to Determine the Sex of a Chick Embryo. 1) To see if there are exosomes present in amniotic fluid of a chick embryo; 2) Identify exosomse by their phenotypes and biochemical composions.