The Analytical Core (Core B) will be organized to provide high quality, reproducible, low cost, automated high-throughput robotic analytical services to the four projects as well as the Animal Core (Core C). The core will be managed and staffed with experts who have demonstrated an ability to develop, engineer, and implement the assays necessary to support the proposed activities of the Analytical Core. The Analytical core will be organized around 4 major services/specific aims.
Specific Aim 1 : To provide high quality, reproducible automated assays to support Projects 1-3 and the Animal Core.
Specific Aim 2 : To provide genotyping services for projects #1 and #2. We will test for the presence of SNPs in GRK4, angiotensin converting enzyme (ACE l/D), aldosterone synthase, angiotensinogen, and ATIR using our previously published method {Bengra, 2002, 12446468} with a data reducfion method (Moore, Pub Med ID 12108579). Additional SNPs will be added as needs arise.
Specific Aim 3 : To provide cultured human renal proximal tubule cells for projects #1and #3 using a novel patent pending 3D cell culture method. We will also use our method to isolate individual renal proximal tubular cells (RPTCs) from fresh specimens of human urine in order to provide greater genetic diversity for studying human physiology, to correlate in vitro cellular responses to in vivo responses in the same patients as studied in Project 2.
Specific Aim 4 : To provide cellular imaging services to support projects #1 and #3. In addition, in support of project #2, we will perform human kidney slice culture and subsequent confocal analysis of DIR and DSR localization.

Public Health Relevance

Core B will develop and use novel analytical techniques to support the needs of the PPG, and to conduct its own hypothesis driven research. Undoubtedly, Core B will also publish its novel analytical developments in the form of patents and original publications.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Program Projects (P01)
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Heart, Lung, and Blood Initial Review Group (HLBP)
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University of Virginia
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Armando, Ines; Konkalmatt, Prasad; Felder, Robin A et al. (2015) The renal dopaminergic system: novel diagnostic and therapeutic approaches in hypertension and kidney disease. Transl Res 165:505-11
Lee, Hewang; Abe, Yoshifusa; Lee, Icksoo et al. (2014) Increased mitochondrial activity in renal proximal tubule cells from young spontaneously hypertensive rats. Kidney Int 85:561-9
Gildea, John J; Shah, Ishan T; Van Sciver, Robert E et al. (2014) The cooperative roles of the dopamine receptors, D1R and D5R, on the regulation of renal sodium transport. Kidney Int 86:118-26
Yu, Peiying; Sun, Min; Villar, Van Anthony M et al. (2014) Differential dopamine receptor subtype regulation of adenylyl cyclases in lipid rafts in human embryonic kidney and renal proximal tubule cells. Cell Signal 26:2521-9
Chen, Ken; Fu, Chunjiang; Chen, Caiyu et al. (2014) Role of GRK4 in the regulation of arterial AT1 receptor in hypertension. Hypertension 63:289-96
Gildea, John J; Seaton, Joscelyn E; Victor, Ken G et al. (2014) Exosomal transfer from human renal proximal tubule cells to distal tubule and collecting duct cells. Clin Biochem 47:89-94
Yang, Yu; Cuevas, Santiago; Yang, Sufei et al. (2014) Sestrin2 decreases renal oxidative stress, lowers blood pressure, and mediates dopamine D2 receptor-induced inhibition of reactive oxygen species production. Hypertension 64:825-32
Jiang, Xiaoliang; Konkalmatt, Prasad; Yang, Yu et al. (2014) Single-nucleotide polymorphisms of the dopamine D2 receptor increase inflammation and fibrosis in human renal proximal tubule cells. Hypertension 63:e74-80
Yu, Peiying; Han, Weixing; Villar, Van Anthony M et al. (2014) Unique role of NADPH oxidase 5 in oxidative stress in human renal proximal tubule cells. Redox Biol 2:570-9
Armando, Ines; Villar, Van Anthony M; Jones, John E et al. (2014) Dopamine D3 receptor inhibits the ubiquitin-specific peptidase 48 to promote NHE3 degradation. FASEB J 28:1422-34

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