*2.3. Surgical and Experimental Procedure*

The transplant procedure used in these experiments is shown in Figure 1. In the BN donor rat, after double ligation of the aorta, ligation of the right renal artery and vein, and surgical section of the left renal vein, the left rat kidney was perfused in situ using 5 mL of room temperature UW Solution (over a 15 s period). The perfusate was either UW Solution alone (for the "0 h" and the "20 h No Treatment" groups), or UW Solution to which crystalline PrC-210, to achieve 30 mM [17], had been added, dissolved immediately, and then pH adjusted to the starting UW Solution pH of 7.4 by adding 0.0619 μL 5N NaOH per μmol of PrC-210 HCL salt (FW: 220). The half-life of PrC-210 thiol (active form) is approximately 3.5 h in physiologic pH solutions such as UW Solution and human blood [14]. Following in situ perfusion, the left BN kidney was surgically removed and then sutured by blunt anastomosis of vessels and ureter into the vacated left kidney site of the LEW recipient rat. The right LEW kidney was ligated and removed immediately before. Five minutes after surgical closure of the LEW rat, the rat received a systemic PrC-210 dose (121 ug PrC-210 HCl per gm body weight, which equals 0.24 X Maximum Tolerated Dose) by intraperitoneal injection. As shown in the Figure 1 schematic, the rat also received intraperitoneal injection doses of PrC-210 (0.24 MTD) at +4 h and +8 h following the transplant. Rats were euthanized at +20 h following transplant, and kidneys and plasma samples were collected for analysis. There were a minimum of five rats in each treatment group.
