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Ischemia reperfusion injury (IRI) contributes significantly to posttransplant graft dysfunction. An emphasis, therefore, has been directed toward the identification of novel renoprotective agents. In this study, the renoprotective effect of tetrodotoxin (TTX) alone, or in combination with a thromboxane synthetase inhibitor (OKY-046), was investigated in a 60-min warm ischemia, 72-h reperfusion, IRI rodent model. Unilateral nephrectomized rats were treated with the test vehicle alone, 1, 2, or 4 microgram/kg of TTX or 2 mg/kg of OKY-046 intravenously, either 15 min pre- or postischemia, or 2 microgram/kg TTX administered simultaneously with OKY-046 (2 mg/kg), following the ischemic interval. Baseline, 24, and 72 h mean plasma creatinine (Cr) and urea nitrogen (BUN) were compared. Maximal renoprotection was demonstrated by significantly improved 72-h Cr and BUN levels with the 2 microgram/kg of TTX or with 2 mg/kg of OKY-046, each administered after ischemia (ischemic control Cr = 8. 01 +/- 1.07 mg/dl vs TTX = 3.84 +/- 0.80 mg/dl, P = 0.008; vs OKY-046 = 4.0 +/- 1.5, P + 0.008; ischemic control BUN = 241.3 mg/dl +/- 32.8 vs TTX = 85.7 mg/dl +/- 18.7, P < 0.008; vs OKY-046 = 52.6 +/- 22.5, P = 0.008). The combination therapy utilizing TTX with OKY-046 resulted in reduced animal survival, demonstrating no renoprotection as measured with the biochemical parameters. These results support the renoprotective effects of TTX in a severe, rodent IRI model. The exact mechanism of action, as well as the therapeutic potential of TTX in preservation/transplantation, warrants further study., (Copyright 1999 Academic Press.)

Free radical mediated lipid peroxidation (LPO) has been implicated in the pathogenesis of ischemic-reperfusion injury (IRI). To address the renoprotective effect(s) of LPO inhibition, the efficacy of the 21 aminosteroid U74389G was evaluated in three IRI models. In Model 1 51 unilateral nephrectomized rats that underwent 60 min of warm ischemia followed by a 72-hr reperfusion interval were treated with the test vehicle only, or 3, 6, or 12 mg/kg of U74389G intravenously, 5 min pre- or postischemia. In Model 2 Sprague-Dawley rats underwent sham operation (n=9), or 45 min of warm ischemia and 10 min of reperfusion with U74389G (6 mg/kg; n=10) or test vehicle only (n=10) administered intravenously over 10 min beginning 5 min prior to clamp release. After reperfusion, LPO was determined by assay of snap frozen tissue for thiobarbituric acid (TBA) concentrations (nmol/g tissue weight). In Model 3 domestic lean maid pigs (14-18 kg) underwent left nephrectomy with 30 min of warm ischemia, Collins C-4 flush, and 24 hr of cold storage preservation. Heterotopic autotransplantation and immediate contralateral nephrectomy was then performed in Group A-nonischemic controls (n=4), Group B-ischemic controls (n=5), and Group C-U74389G (6 mg/kg) administered preischemia and at autotransplantation (n=5). In Model 1 maximal renoprotection was demonstrated with the 6 mg/kg dose of U74389G administered after ischemia (ischemic control 72-hr serum creatinine (Cr) = 8.01+/-1.1 mg% vs. 3.32+/-0.96 mg%; ischemic control creatinine clearance = 0.069+/-0.03 ml/min vs. 0.206+/-0.04 ml/min; P<0.05). In Model 2 TBA levels were significantly lower in U74389G treated animals (88.5+/-10.0 vs. ischemic controls = 296.8+/-81.4; P=0.02). In Model 3 graft survivals were 100%, 0%, and 60% respectively. Peak Cr and BUN (mg%) were significantly greater in Group C vs. Group A, (Group A Cr = 8.59+/-0.63 vs. Group C = 12.8+/-1.01; Group A BUN = 64.1+/-2.73 vs. Group C = 104.9+/-12.21)--however, by day 10, thee were no significant differences in renal function: (Group A Cr = 2.15+/-0.3 vs. Group C = 2.10+/-0.06; Group A BUN = 27.0+/-6.0 vs. Group C = 31.1+/-6.4). These results support the beneficial effects of LPO inhibitors in models of ischemia-reperfusion, as well as preservation/transplantation, and suggest that this renoprotection correlates with decreased membrane lipid peroxidation.

The efficacies of two scintigraphic and two sonographic techniques and resultant index values, as markers of renal allograft pathology, were assessed. Index values of 183 combined scintigraphic and sonographic examinations in 47 graft recipients were compared to the pathological diagnosis of transplant biopsies and subsequent clinical outcome. All recipients were studied with baseline imaging techniques postoperatively, again when indicated by predefined clinical criteria, and prior to graft biopsy. The scintigraphic technique involved the calculation of indices of thrombotic activity and cortical graft perfusion. Ultrasound involved determination of the Doppler resistance index of Pourcelot and estimations of graft volume from real time images. A decreased cortical perfusion index was, overall, the most sensitive index of acute or chronic graft pathology, but it lacked specificity. Increased thrombotic and resistance indices were 96% and 86% sensitive for acute vascular rejection and were 82% and 76% specific. Jointly increased thrombotic and resistance indices improved the specificity for acute vascular rejection to 98%. An increase in graft volume of more than 50% over stable values was 100% sensitive and 92% specific for acute interstitial rejection, and 95% specific when paired with a normal thrombotic index. A marked increase in the thrombotic index was 100% sensitive for cyclosporine-induced thrombotic microangiopathy, but only 49% specific. The specificity of a markedly increased thrombotic index for thrombotic microangiopathy improved to 93% when the Doppler resistance index remained normal or was only marginally elevated. None of the scintigraphic or ultrasound indices were helpful for the diagnosis of acute tubular necrosis, chronic rejection, recurrent glomerulopathy, or graft infection.

To determine the value of scintigraphy to detect posttransplantation complications of the allografted pancreas, we retrospectively reviewed 209 scintigrams obtained with 99mTc-sulfur colloid (99mTc-SC) and 99mTc-glucoheptonate (99mTc-GH). The scintigraphic studies were performed in 37 recipients of simultaneous renal and pancreatic allografts harvested from the same donor. 99mTc-SC was used as an indicator of thrombotic vasculitis; pancreatic perfusion and blood-pool parameters were monitored with 99mTc-GH. In 11 of the 37 recipients, scintigraphic abnormalities suggested posttransplantation infarction. Recurrent episodes of acute rejection of the pancreatic allograft, which always coincided with acute rejection of the renal allograft, were monitored in 24 recipients. Rejection-induced ischemic pancreatitis was suggested in 12 of the 24 recipients and persisted in 10 recipients for several weeks after improvement of renal allograft rejection. Pancreatic atrophy was suggested scintigraphically in 16 of the 24 recipients with recurrent episodes of rejection. Spontaneous pancreatic-duct obstruction and obstructive pancreatitis were associated with a scintigraphic pattern similar to that of rejection-induced ischemic pancreatitis. We concluded that the specific radionuclides used in this series are useful for the surveillance and assessment of posttransplantation pancreatic infarction, acute rejection, pancreatitis, and atrophy.

The isolated perfused organ is more sensitive to the toxicity of oxygen since hypothermia reduces the activities of enzymes responsible for minimizing oxygen toxicity. To protect the organ under these conditions reducing agents must be added to the perfusate. Quantitation of the resulting reduction is best obtained by measurement of the oxidation-reduction potential of the perfusate. A device was designed for this purpose and, by electrochemical principle, controlled reduction of the oxidized form of the oxidation-reduction couple was affected. Kidneys were perfused with cryoprecipitated plasma. With the electrochemical cell in the circuit, the oxidation-reduction potential of the perfusate was adjusted by the addition of ascorbic acid and glutathione and the cell was driven by a battery-powered potentiostat. Kidneys subjected to 60 minutes of warm ischemia had optimal survival at -20 mV. Preservation for six days in a monitored group had no survivors, whereas kidneys with oxidation-reduction support maintained life. Optimal oxidation-reduction support maintained life. Optimal oxidation-reduction was at or near -17 mV. These data show a requirement of an optimal oxidation-reduction potential to reverse warm ischemia damage and to prolong the period of ex vivo preservation of isolated perfused organ.