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The results were expressed as picomoles of CO per milligram of tissue. Human kidney grafts that were deemed nontransplantable were obtained from our local OPO and used for control and CO-UW static storage experiments, which was approved by the Committee for Oversight of Research Involving the Dead (CORID) of the University of Pittsburgh and the Center for Organ Recovery & Education (CORE). As soon as the nontransplantable kidney graft became available for research, a baseline sample was obtained. A catheter was inserted into the upper renal arterial branch, and 100?ml CO-UW (10%) was perfused, while the lower part of the kidney was perfused with 100?ml control UW. The kidney graft was split into upper and lower portions, and stored http://www.selleck.cn/products/s-gsk1349572.html for 24?h in CO-UW and control UW solution respectively. After 24?h cold static http://www.selleckchem.com/products/ly2157299.html storage, kidney graft samples were obtained for CO contents, histopathology, Western blot, and real time RT-PCR. In addition, explants were obtained by precisely cutting kidney grafts into 2.0?��?5.0?��?5.0?mm cubes using straight adjustable tissue cutting channel TM-S-9 (Braintree Scientific, Inc. Braintree, MA, USA), and incubated in 12-well plates with 1.0?ml of Dulbecco��s Modified Eagle��s Medium, supplemented with 4.5?g/l glucose, 2?mM, L-glutamine, 5% FBS, 0.4?IE/ml insulin, 20?mM HEPES, and 50?��g/ml gentamycin. Plates were cultured for 3, 6, and 12?h at 37?��C under gentle shaking in a conventional 5% CO2 incubator [21]. In this human kidney study, we used one http://www.selleckchem.com/products/z-vad-fmk.html kidney graft for both control UW and CO-UW to have the same baseline values, as histopathological findings were different between two kidneys from the same donor. Proper perfusion with control UW and CO-UW was confirmed by measuring tissue CO levels. Data are represented as the mean?��?SD. Comparisons between groups at different time points were performed using Student��s t-test or anova. Graft survivals were plotted with the Kaplan�CMeier method, and the difference among groups was analyzed using the log-rank test. Differences were considered significant at a P-value?