Gas chromatographic determination of residual ethylene oxide in the endotracheal tube

An accurate and simplified method has been developed for determination of residual ethylene oxide (EO) in the endotracheal tube using gas chromatography with a head space sampler. The endotracheal tube made of polyvinyl chloride was sterilized with EO and aerated. Residual EO after aeration was 6630 +/- 1915 ppm (mean +/- SD). It decreased to 72 +/- 46 ppm on 7th day. The endotracheal tube should be left unused for more than 7 days after sterilization and aeration. The safety level of residual EO in the endotracheal tube by U.S.FDA regulation is less than 250 ppm. The concentration of residual EO should be lower than 250 ppm, if this rule is observed.     

Rinsing morselized allografts improves bone and tissue ingrowth

Bone defects in revision hip surgery can be reconstructed with impacted morselized bone grafts. Rinsing these trabecular allografts may enhance graft incorporation by washing out immunogenic factors present in blood, marrow, and fat. However, it has been proposed that impaction of the graft releases biologically active factors, which can provide sufficient activity to stimulate new bone formation. Rinsing before impaction could enhance bone allograft incorporation, but rinsing after impaction could diminish the incorporation process of impacted bone graft. To study the effect of rinsing and impaction of morselized bone grafts on bone ingrowth, a bone chamber study was done in goats. Autografts and allografts were divided into three treatment groups: (A) impacted; (B) rinsed and impacted; and (C) rinsed, impacted, rinsed, and impacted again. Ten goats received three bone chambers in each proximal tibia. The chambers were filled with either allograft or autograft, yielding six different implants per goat. After 6 weeks, histologic analyses were done and bone and tissue ingrowth were measured. New bone and total tissue ingrowth were higher in autografts than in allografts, especially in the nonrinsed group. With rinsing, total tissue ingrowth increased in the allograft group to approach that of autografts. Rinsing after impaction did not additionally alter bone ingrowth. The current findings show that incorporation of allografts can be improved by rinsing the grafts before impaction.     

Ethylene Oxide in Dialyzer Rinsing Fluid: Effect of Rinsing Technique, Dialyzer Storage Time, and Potting Compound

Ethylene oxide (ETO) is recognized as one of the main causes of dialyzer-associated hypersensitivity reactions. We studied the amount of ETO in the rinsing fluid of ETO-sterilized hollow-fiber dialyzers as a function of rinsing technique, dialyzer storage time, and the amount of potting compound (known to be an ETO reservoir) in the dialyzer. The results suggested that the initial 500 ml of rinsing fluid removes much of the residual ETO in the dialyzer. Ethylene oxide extraction was enhanced substantially by rinsing at 37 degrees C versus 5 degrees C. However, considerable amounts of ETO remained in the dialyzer after an initial 500 ml rinse, some of which could be removed by rinsing with an additional 1,500 ml. High concentrations of ETO were measured in fluid that had been recirculated through the dialyzer for 10 min or longer and in fluid that had been allowed to remain in the dialyzer for 10 min under zero-flow conditions. The amount of ETO in the rinsing fluid decreased markedly as the dialyzer storage time was increased from 4 to 8 weeks and in dialyzers in which a portion of the potting compound had been replaced with a polycarbonate ring. Our results suggest that the dose of ETO administered to the patient at the outset of dialysis can be minimized by rinsing the dialyzer with 2 L of fluid at 37 degrees C and by avoiding administration of rinsing fluid that has been allowed to remain in contact with the dialyzer for more than several minutes. Use of a long storage interval and use of dialyzers containing reduced amounts of potting material will also reduce the ETO load.     

Use of an amphoteric lavage solution for emergency treatment of eye burns. First animal type experimental clinical considerations

Purpose: Severe eye burns occur rarely, but are related to a poor prognosis in rehabilitation. As emergency treatment has been identified as decisive factor for the prognosis of eye burns, new first aid rinsing solutions should be considered carefully in their clinical action. In a first approach, the new drug Diphoterine^® was subjected to a comparison with saline solution to evaluate the effects in a model of severe eye burns.

Methods: In a double-masked experiment 16 rabbits underwent a severe eye burn of one cornea followed by immediate rinsing with 0.9% sodium–chlorine solution (n=8) or Diphoterine^® (n=8). During 16 days after burn, an irrigation therapy with 0.9% saline solution three times daily 160 ml was applied in both groups following the recommendation of prolonged irrigation therapy performed in our clinic. In a similar setup, 16 eyes were subjected alkali burns with measurements of aqueous humor pH within 30 s after burn and after a period of 5 min rinsing with 500 ml saline 0.9% or Diphoterine^®, respectively.

Results: The result of the severe eye burn with an opaque cornea was similar in both groups. During rinsing no fibrin precipitates occurred in the Diphoterine^® rinsed group whereas this was detectable in all eyes rinsed with saline solution. After 16 days there was no difference between both groups indicating no harmful effect of Diphoterine^® as emergency treatment compared to saline 0.9%. After 30 s of burn with 1N NaOH and rinsing with 500 ml of the specified solutions the anterior chamber pH was 10±0 in the saline group and 9.35±0.3 in the Diphoterine^® group showing efficacy of the buffering capacity of Diphoterine^®.

Conclusion: Diphoterine^® proves to be efficient in the primary treatment of burns. The anterior chamber pH could be lowered by 5 min of rinsing. No harmful effects of Diphoterine^® could be observed compared to rinsing with saline solution in the course of an severe alkali burn of the cornea.     

内生式置换液预冲透析器的应用效果观察

目的探讨内生式置换液代替无菌生理盐水冲洗透析器的临床应用效果。方法将 1 0 0 0套血路管、透析器随机分为实验组和观察组各 5 0 0套 ,实验组采用GAMBROAK 2 0 0ULTRA血液净化机产生的内生式置换液进行透析前预冲洗 ,对照组则用无菌生理盐水进行冲洗。对冲洗后的透析器进行细菌培养和内毒素检测。结果两组透析器均未检出细菌和内毒素 ;实验组透析器冲洗、排气能一次完成 ,冲管工序简单、节省人力、费用少 ,与对照组比较 ,差异有显著性意义 (P <0 .0 5 ,P <0 .0 1 )。结论采用内生式置换液对血路管、透析器进行冲洗 ,安全 ,成本低     

Hemodynamic and morphological changes in the dog kidney after injection of 5% ethanolamine oleate into the superior vena cava

The effect of the sclerosant 5% ethanolamine oleate (EO) on renal circulation was evaluated in 20 mongrel dogs into which we injected 5% EO (0.5 ml/kg) into the superior vena cava. There was a marked hemolysis and a significant decrease in creatinine clearance from 104.4 +/- 17.1 (mean +/- SD) to 40.7 +/- 5.0 ml/min (p less than 0.01) at 120 min. The renal arterial blood flow (RAF) decreased biphasically; from 75.3 +/- 13.1 to 9.8 +/- 9.3 ml/min during an average of 2.3 min, immediately after the injection (p less than 0.01) and gradually decreased after reaching the pretreatment level to 43.2 +/- 8.6 ml/min at 120 min (p less than 0.01). Cardiac output significantly decreased from 1.86 +/- 0.08 to 1.54 +/- 0.08 l/min (p less than 0.01). Tubular necrosis was histologically evidenced in the tissues examined at 6 h after the injection of EO. Biphasic decrease in renal arterial blood flow can be explained by the possible occurrence of spasm of the peripheral renal arteries in the acute phase and tubular necrosis in the late phase. This study suggests to us that the tubular necrosis induced by decreases in RAF as well as hemolytic nephropathy play a significant role in cases of renal dysfunction following the endoscopic injection of 5% EO to sclerose esophageal varices.     

Long-term survival of skin allografts in rats treated with topical cyclosporine

The use of topical cyclosporine (CsA) was studied in skin allografts from Buffalo to Lewis rats. CsA prepared in olive oil and dimethyl sulfoxide was administered in various dosages topically on allografts. Untreated allografts were rejected in 7.4 +/- 1.1 days but survived for 18.6 +/- 0.9, 29.3 +/- 1.8, or 40.6 +/- 2.2 days after 10, 20, or 28 days of topical CsA treatment (10 mg/rat/day), respectively. Long-term graft survival (greater than 100 days) was seen with continuous CsA treatment at 10 mg/rat/day, 10 mg/rat/2 days, and 5 mg/rat/day, as compared with rejection in 13.1 +/- 2.3 and 8.9 +/- 0.9 days with CsA 10 mg/rat/3 days and 5 mg/rat/2 days, respectively. The therapeutic blood level of CsA ranged from 250 to 500 ng/ml. Most grafts were rejected when CsA blood levels fell below 200 ng/ml. Direct administration of topical CsA onto the allografts resulted in longer survival compared with those applied on the normal recipient skin 6 cm distal to the allografts, with both high and low doses. Locally high concentrations of CsA in allografts may play an important role in prolongation of graft survival. Minimal cell infiltration and loss of hair follicles were the main histological features in long-surviving allografts (greater than 120 days).     

A modified University of Wisconsin preservation solution with high-NA+ low-K+ content reduces reperfusion injury of the pig kidney graft

BACKGROUND: Ischemia-reperfusion injury has been associated with both early and late effects on allografts in the form of delayed graft function and decreased graft survival. Recent studies demonstrated that functional parameters were influenced by cold storage conditions and particularly the ratio of Na+:K+ of the preservation solution. METHODS: We have extended this study to examine whether the high-Na+ low-K+ formulation of Belzer's solution (HEH) was efficient in an autotransplanted pig kidney model when compared with the classical low-Na+ high-K+ University of Wisconsin solution and the new high-Na+ low-K+ Celsior solution. Kidneys were harvested, cold flushed, and preserved for 24, 48, or 72 hr with HEH, Celsior solution, or University of Wisconsin solution and autotransplanted. Renal function was determined on days 1, 3, 7, and 14, and at 4 to 16 weeks after autotransplantation. Histologic changes and cell infiltration were assessed on kidney biopsy specimens taken after reperfusion (30-40 min), at days 5 and 14, and at 4 to 5 and 10 to 12 weeks after surgery. Peripheral benzodiazepine receptor (PBR), a structural mitochondrial protein, was also studied. RESULTS: Cold storage in HEH resulted in reduction of delayed graft function and renal damage, with a decrease in interstitial inflammation. HEH reduced interstitial fibrosis, tubular atrophy, and improved PBR expression. CONCLUSION: This study suggests that cold preservation in HEH has a beneficial action in in vivo renal preservation and reduces tubular necrosis, interstitial inflammation, and fibrosis in these groups. In addition, PBR detection was correlated to the level of preservation integrity.     

漂洗对脂肪活性的影响

目的比较不同漂洗液和不同漂洗次数对吸脂术所获得的脂肪组织活性的影响。方法通过肿胀吸脂术获得脂肪组织后,分别用生理盐水、平衡液各漂洗0、1、3、5、10次后,用葡萄糖转移实验检测其活性。结果生理盐水组脂肪组织的葡萄糖转移量大于平衡液组,差异具有统计学意义(P<0.05);两组中,漂洗0和1次时的葡萄糖转移量明显低于漂洗3次(P<0.01);生理盐水组中,漂洗3次和5次时的葡萄糖转移量无明显差异(P>0.05),但漂洗10次时的葡萄糖转移量明显低于漂洗3次和5次(P<0.05);平衡液组中,漂洗3次、5次和10次时的葡萄糖转移量无明显差异(P>0.05)。结论通过漂洗可以提高吸脂术所获得的脂肪组织的活性。本实验中,生理盐水的漂洗效果优于平衡液,以漂洗3次为佳。     

Enhancement of hypothermic heart preservation with fructose 1, 6-diphosphate.

BACKGROUND: We hypothesized that the addition of fructose 1, 6-diphosphate (FDP) to a hypothermic heart preservation solution could improve metabolic recovery because it has several beneficial effects. MATERIALS AND METHODS: Twenty adult Sprague-Dawley rats were used to study hypothermic heart preservation. The hearts were removed under general anesthesia and preserved at 4 degrees C in Euro-Collins solution (30 ml/kg) for 8 h. In the study group (N = 10), FDP (5 mM) was added to the Euro-Collins solution. In the control group (N = 10), no FDP was added. Heart function was studied after preservation using a working heart model. The ability of various concentrations of fructose 1,6-phosphate to passively diffuse through an egg phosphatidylcholine multilamellar vesicle (MLV) membrane bilayer was examined. RESULTS: Cardiac output ranged from 17.0 +/- 1.9 to 24.9 +/- 1.6 ml/min in the study group vs 2.0 +/- 1.0-12.3 +/- 1.7 ml/min for controls, average aortic flow was 10. 8 +/- 1.4 ml/min in the study group vs -1.3 +/- 1.6 ml/min for controls, and maximum LV generated power was 22.8 +/- 1.7 J/min vs 10.1 +/- 1.6 J/min for controls. Coronary flow, left ventricular stroke volume and stroke work, and myocardial oxygen consumption were much higher in the study group than in the control group. Coronary vascular resistance was lower in the study group than in the control group. Electron microscopic study indicated that many myocytes displayed patches of swollen mitochondria in the control group, but was rarely observed in the study group. The addition of 50 mM FDP caused substantial changes in MLV permeability. No dose of sucrose buffers outside the vesicles resulted in a significant changes of MLV permeability. CONCLUSIONS: Our results indicate that the addition of FDP to Euro-Collins solution significantly improves hypothermic rat heart preservation, and FDP appeared to cross the membrane bilayer.     

Liver preservation prior to transplantation: Past,present, and future

Since Dr. Thomas Starzl performed the first series of successful liver transplants(LTs), important advances have been made in immunosuppression, operative techniques, and postoperative care. In 1988, Belzer's group reported the first successful LT using the University of Wisconsin preservation solution(UW).Since then, UW has replaced EuroCollins solution and allowed prolonged and safer preservation of liver, kidney, and pancreas allografts, thus contributing to the improvement of transplant outcomes. Although UW is still considered the standard of care in the United States and in several countries worldwide, a recent meta-analysis revealed similar LT outcomes among UW, Celsior solution, and the Institut Georges Lopez-1 preservation solution, which were slightly superior to those obtained with histidine-tryptophan-ketoglutarate preservation solution.Dynamic preservation has been recently developed, and liver allografts are preserved mainly through the following methods: hypothermic machine perfusion, normothermic machine perfusion, and subnormothermic machine perfusion. Their use has the potential advantage of improving clinical results in LT involving extended criteria donor allografts. Although associated with increased costs, techniques employing machine perfusion of liver allografts have been considered clinically feasible. This editorial focuses on recent advances and future perspectives in liver allograft preservation.     

Prevention of spinal anaesthesia-induced hypotension in the elderly: i.m. methoxamine or combined hetastarch and crystalloid

We have compared two methods of reducing hypotension during spinal anaesthesia in elderly patients, 6% hetastarch and crystalloid or methoxamine 10 mg i.m., in terms of haemodynamic stability and requirements for additional vasopressors. Sixty-two patients (aged 60- 97 yr) undergoing surgical fixation of fractured neck of femur were allocated randomly to receive 6% hetastarch (Hespan) 500 ml followed by Hartmann's solution 500 ml (group HS, n = 32) or a bolus injection of methoxamine 10 mg i.m. (group MX, n = 30), 10 min before induction of spinal anaesthesia with 0.5% hyperbaric bupivacaine 2.25-3.0 ml. Arterial pressure was measured non-invasively by an oscillotonometer at 2-min intervals from 0 to 40 min and at 5-min intervals thereafter. Methoxamine 2 mg i.v. was given if systolic arterial pressure (SAP) decreased to < 100 mm Hg. Hypotension was defined as a 25% decrease from baseline SAP or mean arterial pressure (MAP). Patient data, sensory level and blood loss were similar in the two groups. SAP and MAP increased initially from baseline until induction of spinal anaesthesia and then decreased for 30 min in both groups, but remained higher in group MX (P < 0.05). Heart rate (HR) decreased from baseline in group MX (P < 0.05) and was less than in group HS at all times from 2 to 60 min (P < 0.01). The incidence of SAP hypotension (47% vs 75%; P = 0.03, odds ratio (OR) = 3.43) and MAP hypotension (47% vs 67%; P = 0.09, OR = 2.51) was less in group MX than in group HS. Requirements for rescue methoxamine i.v. (27% vs 53%, P = 0.04, OR = 3.11) was less in group MX than in group HS but the dose of rescue methoxamine given (mean 6.3 (95% confidence intervals 3.0-9.6) vs 8.9 (5.6-12.2) mg) and time to onset of hypotension (20.7 (14.5-26.7) vs 17.3 (11.4-23.1) min) were similar in groups MX and HS, respectively. We conclude that methoxamine 10 mg i.m., given 10 min before induction of spinal anaesthesia in normovolaemic elderly patients, reduced subsequent SAP and MAP hypotension, HR and requirements for rescue vasopressor therapy compared with a combination of 6% hetastarch 500 ml and crystalloid 500 ml. The previously reported benefit of such volume administration may not extend to the elderly.      

ELISA法检测组织工程产品中残留牛血清蛋白含量

目的探讨定量检测牛血清白蛋白（Bull serum albumin，BSA）酶联免疫试剂盒用于组织工程医疗产品（Medical products of tissue engineering，TEMPs）残留BSA检测的适用性，以及清洗程序对产品BSA残留量的影响。材料和方法检测了三种组织工程医疗产品（组织工程骨、组织工程肌腱以及组织工程角膜贴片）中BSA的残留量，并比较了经不同清洗程序清洗后角膜贴片中残留BSA含量。结果（1）组织工程骨、组织工程肌腱以及组织工程角膜贴片中BSA残留量分别为31．8ng／ml，4．9ng／ml和低于2．5ng／ml，且测定值与其稀释倍数呈正相关；（2）生理盐水及无血清的MEM培养基平均OD值与试剂盒中零浓度标准品接近．表明生理盐水或无血清MEM作为样品浸提介质对最终的实验结果无明显影响；（3）样品复孔比均小于1．24，符合试剂盒有效性要求。经清洗程序2处理后，角膜贴片中残留BSA较清洗程序1明显减少。结论（1）该试剂盒适用于组织工程医疗产品中BSA残留量的检测。（2）对于组织工程医疗产品而言，不同的清洗程序对BSA的残留量有非常明显的影响。生产商应根据其预期用途制定有效的清洗程序，并在说明书中给予足够的阐述。     

Viruses adsorbed on musculoskeletal allografts are inactivated by terminal ethylene oxide disinfection.

In 1987 it was anticipated that unsterilized tissues would transmit virus diseases such as hepatitis and HIV-1 from infected donors so a freeze-drying process for musculoskeletal tissue was developed to include terminal ethylene oxide (EO) exposure for 14 h. We found no studies of EO efficacy when viruses were associated with human allografts so we studied the antiviral effect of terminal EO disinfection using all but the final freeze-drying phase of this clinical processing protocol (CPP). Specifically we looked at EO inactivation of HIV-1, a human hepatitis B surrogate and test viruses known to be highly resistant to disinfecting agents, including irradiation. Freeze-drying, ordinarily required after EO disinfection and part of the CPP, was not done. Suspensions of HIV-1, Bovine viral diarrhea, Reovirus type 3, Duck hepatitis B, Poliomyelitis and Canine parvovirus were adsorbed on glass, demineralized bone powder, and preprocessed strips of femoral cortex, iliac wedges, cancellous blocks and patellar bone-tendon-bone preparations and subjected to EO disinfection. Test viruses were inactivated at the end of 7 h of EO disinfection, providing a safety factor in the CPP of at least 100%. Because allografts can transmit viruses, terminal EO disinfection should provide safer musculoskeletal allografts than non-disinfected tissues or those irradiated with a standard irradiation dose. New spontaneously appearing viruses would probably be inactivated with this terminal EO disinfection but they and viral bioweapons will require individual validation to assure viral inactivation.     

Intraoperative total serum calcium levels,unlike intraoperative intact PTH levels,do not correlate with cure of hyperparathyroidism

BACKGROUND: Intraoperative intact parathyroid hormone (iPTH) monitoring is useful in the operative management of hyperparathyroidism. Recent studies suggest that measurement of intraoperative total serum calcium (TSC) levels may be a more cost effective and readily available method of intraoperative guidance during neck dissection than iPTH levels, the gold standard. We compared the accuracy of intraoperative TSC to iPTH in predicting surgical cure during parathyroidectomy. PATIENTS AND METHODS: From September 1, 2001 to October 31, 2002, 88 parathyroidectomies were performed. iPTH and TSC were measured at the start of the operation, and at 5 and 10 min after gland removal. Data were compared, and trends were analyzed with respect to removal of abnormal parathyroid tissue as confirmed by pathology. One-way analysis of variance was used to determine if decreases in TSC were significant. RESULTS: The mean baseline iPTH level (418 +/- 610 pg/ml) dropped by 70% 5 min after removal of the abnormal glands (86 +/- 102 pg/ml) and by 85% at 10 min (39 +/- 39 pg/ml). The mean baseline TSC level (10.0 +/- 0.8 mg/dl) dropped by 4% at 5 min after removal of the abnormal glands (9.6 +/- 0.9 mg/dl) and remained at 4% at 10 min (9.6 +/- 0.8 mg/dl). iPTH dropped by > or =50% in 73 patients (83%) at 5 min and in 87 patients (99%) at 10 min after gland resection. TSC decreased below baseline at 5 min and remained below baseline at 10 min in only 47 patients (54%). In the remaining patients, intraoperative TSC changes were less predictable and did not respond consistently to resection of abnormal glands. CONCLUSIONS: The decreases in TSC during parathyroidectomy, if present, are minimal. Unlike iPTH levels, TSC levels do not consistently decrease at 5 and 10 min after gland resection. While attractive in terms of cost and availability, intraoperative TSC levels are not clinically reliable in confirming removal of abnormal parathyroid tissue.     

Progressive coronary vasoconstriction during 25 hours of myocardial preservation in vitro impairs functional capacity following preservation

We have developed perfusion techniques for preserving rat hearts for 25 h and have quantified haemodynamic function after preservation to establish the relation between coronary vascular resistance during preservation and the quality of postpreservation pump function. Thirteen rat hearts underwent hypothermic (8°C), low-pressure (15 mmHg) perfusion with an hyperosmotic (385 mOsm/l) crystalloid preservation buffer for 25 h. During this period, the coronary flow rate decreased from 1.12±0.28 ml/min to 0.87±0.12 ml/min (±SD). Following the preservation period, the quality of pump function was tested in the isolated working heart model. At a fixed value of left atrial pressure (15 mmHg), the afterload was increased stepwise (5 mmHg) from 45 mmHg to 70 mmHg, making use of a Starling resistor in series with an air compliance. Each afterload step was maintained for 5 min to obtain stable readings of cardiac output and coronary flow. These measurements were compared with those from a control group of 10 rat hearts undergoing the same test protocol for haemodynamic function without previous preservation. The 13 hearts which underwent 25 h preservation had subnormal haemodynamic function: cardiac output was 50%±4% compared to 10 control hearts. If preserved hearts were divided into two groups based on coronary vascular resistance measured at the end of the preservation period lower than 18 mmHg · min per ml (group 1), and higher than or equal to 18 mmHg · min per ml (group 2), it appeared that the haemodynamic function of group 2 hearts was about half that of group 1 hearts. It is concluded that the haemodynamic function of rat hearts following preservation is negatively affected by coronary vasoconstriction occurring during preservation.     

剖宫产术中输注葡萄糖对产妇及胎儿脐血血糖,胰岛素的影响

作者对剖宫产时产妇分娩前输入5g或25g糖时,发现在分娩前输入25g糖的产妇及胎儿血糖、胰岛素比输入5g糖的产妇及胎儿明显增高。     

Efficacy of Charcoal Hemoperfusion in Paraquat Poisoning

We studied the efficiency of in vitro and in vivo charcoal hemoperfusion for the removal of paraquat. At a flow rate of 200 ml/min, 93–99% of paraquat in 4 L of solution (5, 10, 100 ppm) was removed in less than 160 min. The disappearance half-time was 16 min 10 sec. At 100 ml/min, it was 49 min 30 sec.
Of 23 paraquat poisoning cases, 15 patients underwent hemoperfusion (HP) of which 10 died of respiratory failure within 28 days (7.6 ± 2.9) and 5 survived without pulmonary complications. Of eight patients who did not receive HP, six died of respiratory failure within 97 days (33.4 ± 18.8), even when their general condition was good upon admission. Two who vomited the ingested paraquat immediately survived without HP, and their paraquat concentration in urine was less than one ppm on admission. In one patient, whose paraquat concentration in blood was followed serially, 99% of the paraquat was removed from circulating blood by a single hemoperfusion.
We conclude that charcoal hemoperfusion is effective for the removal of paraquat from blood in vivo and from solution in vitro. Hemoperfusion may also improve survival after paraquat ingestion, but further data are needed.     

Donor dopamine pretreatment inhibits tubulitis in renal allografts subjected to prolonged cold preservation

BACKGROUND: In the present study, we used the Brown-Norway (BN) to Lewis model as a model for acute rejection, to test the hypothesis that dopamine (DA) treatment of BN donors significantly reduces the inflammatory response after renal transplantation. METHODS: BN and Lewis rats (isograft controls) were treated for 24 hr with DA (5 microg/kg/min) or NaCl (0.9%), respectively. After 24 hr of cold storage in University of Wisconsin (UW) solution, renal allografts were orthotopically transplanted into Lewis recipients. All recipients received immunosuppression until they were sacrificed. Allografts were harvested one, three, five, and 10 days after transplantation and analyzed by light microscopy, immunohistochemistry (CD3, major histocompatibility complex [MHC] class II, ED1, P-selectin and intercellular adhesion molecule [ICAM]-1) and by RNase protection assay for cytokine mRNA. RESULTS: Ten days after transplantation Banff tubulitis scores were significantly lower in DA-treated than in NaCl-treated allografts. No significant differences were found in Banff interstitial infiltration scores. The numbers of MHC class II+ and CD3+ cells were significantly decreased in DA-treated animals as assessed by immunohistochemistry. No differences were found in the number of ED1+, P-selectin+, and ICAM-1+ cells. The expression of Ltalpha, tumor necrosis factor, interleukin-1beta, and interleukin-2 mRNA was significantly reduced in DA-treated animals. CONCLUSION: Our data indicate that donor DA treatment significantly inhibits tubulitis in renal allografts subjected to prolonged cold preservation. A reduced number of infiltrating MHC class II+ and CD3+ cells together with decreased cytokine expression could diminish renal scarring, reduce allograft immunogenicity, and hence improve transplantation outcome.     

Local liberation of cytokines during liver preservation

In order to investigate locally produced mediators during the process of organ storage in liver transplantation, we collected the liver preservation solution effluent of 15 transplanted livers and compared it with serum samples taken preoperatively from donor and recipient, as well as 60 min after reperfusion. The mean ischemia time ± SEM was 10 h 10 min ± 53 min. Mean concentrations in University of Wisconsin preservation solution effluent were: interleukin-(IL-)1β 154 ± 77 pg/ml; IL-1 receptor antagonist (IL-1 ra) 1281 ± 309 pg/ml; IL-6 412 ± 90 pg/ml; and for tumor necrosis factor-(TNF-)α 74 ± 21 pg/ml. Cytokine levels in the donors were lower than those detected in the effluent. All measured cytokines showed higher concentrations in the effluent compared to those of the recipient prior to the operation. With respect to a comparison of donor and recipient values, no correlation is evident. Likewise, the ischemic time does not correlate with effluent values. Further development of liver preservation concepts requires information about the state of the graft before reperfusion. Data on cytokine liberation may serve as a helpful tool for the further development of preservation concepts because they enable an estimation of cell activation during preservation. Received: 2 June 1997 Accepted: 10 November 1998