Euro-Collins Solution Research Articles

Effects of Eurocollins Solution as Aortic Flush for the Procurement of Human Pancreas

Belzer solution is considered to be the best preservation media used for pancreas transplantation; however, its high cost accounts for approximately 14.5% of all resources allocated by the Brazilian government toward each pancreatic transplant. The objective of the present study was to test a reduction of Belzer solution during pancreas harvest, thereby lowering procedural cost. The patients received pancreas-kidney transplantations during the period from January 2003 to August 2004. Patients were divided into two groups. Patients assigned to Group A (n=30) received only Belzer solution (2 L through the aorta artery), whereas patients in Group B (n=16) were perfused first with 1 L of Eurocollins solution followed by 1 L of Belzer solution. The two groups were assessed for differences in the following clinical parameters: the need for insulin replacement or antifungal and anticytomegalovirus treatment, pancreatitis, acute cellular rejection, graft vascular thrombosis, fistulas, intra-abdominal collection, graft loss, deaths, pancreatic ischemia time, and average hospitalization time. No statistically significant differences were observed in any of the parameters analyzed (P<0.05). The use of Eurocollins solution, followed by Belzer solution during pancreas harvesting, did not result in differences in graft survival or functionality, postsurgical complications, or patient survival and hospitalization time, when compared to the use of Belzer solution alone. Perfusion with 1 L of Eurocollins solution followed by 1 L of Belzer solution during pancreas harvesting seems to be a simple and efficient alternative for reducing the costs of the harvesting process.

Downregulation of Cytokine Release by Heat Preconditioning of Livers Used for Transplantation in Rats

We evaluated the changes in the levels of released cytokines following heat preconditioning of the livers used in rat liver transplantation. The donor rats in the heat preconditioning (HP) group were subjected to heat preconditioning 48 h before graft harvesting. The liver isografts were preserved in Euro-Collins solution for 8 h, and then transplanted orthotopically. The one-week survival rate of the HP group was significantly better than that of the control (C) group. The serum levels of interleukin-6 and interleukin-10 were significantly lower in the HP group than in the C group. Histological staining revealed that the stagnation of red blood cells and infiltration of neutrocytes were reduced in the HP group. The expression of intercellular adhesion molecule-1 was decreased around the central vein in the HP group, as revealed by immunohistochemistry. These results indicate that heat preconditioning downregulates cytokine release and reduces the frequency of microcirculation disorders.

Calcium channel blocker and renal mitochondrial function in warm renal ischemia

Ions, particularly calcium ions, play an important role in ischemia-reperfusion cell injury. In this study, we investigated the action of verapamil on the mitochondrial function of kidneys submitted to ischemia without blood reperfusion in order to study isolated early and late ischemic effects. 44 rats were submitted to bilateral warm renal ischemia for 30 minutes. The kidneys were then immediately reperfused with saline or Euro-Collins (EC) solution, with and without previous administration of 0.35 mg/kg of verapamil. Mitochondrial function was assessed at the end of renal perfusion and after 24 hours of cold preservation. In kidneys perfused with saline, verapamil allowed a significant early preservation of state III mitochondrial respiration, a result that was no longer evident after 24 hours. In kidneys perfused with EC solution, verapamil did not change state III for either early or late evaluations. Comparison of the groups showed that the results obtained for kidneys perfused with EC were always superior to those obtained for the saline group, except for the initial analysis of kidneys treated with saline and verapamil, which showed results similar to those obtained with EC perfusion alone. Administration of verapamil before warm ischemia provides partial and short-lasting functional protection of the mitochondrial function in kidneys perfused with sodium rich saline. With Euro-Collins solution, verapamil did not show any additional beneficial effect. This fact permits us to conclude that protective action is effective only under conditions that facilitate increased sodium uptake and/or potassium loss.

A perfusionist’s role in lung transplant preservation

The major obstacles of organ transplantation are the shortage of available organs from donors and the limited time an organ remains viable. The scarcity of organs has increased the importance of effective preservation technology. The optimal lung preservation techniques are controversial and in a state of evolution. Recent areas of research include the ideal perfusate solution, the benefit of different pharmacological additives and the best techniques for organ preservation. Our technique for lung preservation has yielded highly successful results. The perfusionist delivers 4-5 L of modified Euro-Collins solution and 500 microg of prostaglandin E1 through a circuit. The designed circuit allows the perfusate to be delivered cold and at a pressure less than 40 mmHg via the use of a manually operated centrifugal pump. In 2002, the operative mortality for all lung transplants was 2.7%. One-year and two-year survival for recipients was 79% and 68% respectively; the national average for one-year survival was around 77% with a 9% mortality. As organ transplant surgery continues to grow, effective organ preservation techniques and technology will expand as well.

Hyperinflation during lung preservation and increased reperfusion injury 1

Reperfusion injury after lung transplantation remains a perplexing and unpredictable problem. Most surgeons preserve the lung inflated, but the amount of inflation that should be used is not well documented. Therefore, we studied the effect of high inflation during organ preservation on lung function during reperfusion. Our hypothesis is that donor lung hyperinflation during storage contributes to early allograft dysfunction during reperfusion. To test our hypothesis we used an isolated, blood-perfused, ventilated rabbit lung model. Group I lungs (control) underwent immediate reperfusion after harvest. Group II lungs (low-inflation, maintained at 6 mmHg airway pressure) and group III lungs (high-inflation, maintained at 20 mmHg airway pressure) were stored for 4 h in 4 degrees C Euro-Collins solution after harvest. All lungs were then reperfused with whole blood for 1 h, and measurements of arterial oxygenation (PO2, mmHg), pulmonary artery pressure (PAP, mmHg), peak inspiratory pressure (PIP, cm H2O), and wet-to-dry weight ratio (WTD) were obtained. Throughout the 1 h reperfusion period group III lungs had significantly lower oxygenation compared to groups I and II. In addition, throughout reperfusion, group III lungs showed significantly higher PAP and PIP compared to group II. WTD did not differ significantly between groups, however, there was a trend toward increased edema in group III. These results indicate that high inflation during cold storage results in acute pulmonary dysfunction. Careful monitoring of airway inflation pressure during storage, especially to prevent hyperinflation, should be maintained in the current practice for lung transplantation.

Assessment of renal ischemia by optical spectroscopy 1,2

No reliable method currently exists for quantifying the degree of warm ischemia in kidney grafts before transplantation. We describe a method for evaluating pretransplant warm ischemia time using optical spectroscopic methods. Lewis rat kidney vascular pedicles were clamped unilaterally in vivo for 0, 5, 10, 20, 30, 60, 90, or 120 min; eight animals were studied at each time point. Injured and contralateral control kidneys were then flushed with Euro-Collins solution, resected, and placed on ice. 335 nm excitation autofluorescence as well as cross-polarized light scattering images were then taken of each injured and control kidney using filters of various wavelengths. The intensity ratio of the injured to normal kidneys was compared to ischemia time. Autofluorescence intensity ratios through a 450-nm filter and light scattering intensity ratios through an 800-nm filter both decreased significantly with increasing ischemia time (P < 0.0001 for each method, one-way analysis of variance). All adjacent and nonadjacent time points between 0 and 90 min were distinguishable using one of these two modalities by Fisher's protected least significant difference. Optical spectroscopic methods correlate with warm ischemia time in kidneys that have been subsequently hypothermically preserved. Further studies are needed to correlate results with physiological damage and posttransplant performance.

Kidney transplantation from non–heart-beating donors: Analysis of organ procurement and outcome

Introduction Most donors in Japan have been non–heart-beating donors (NHBD), so-called “marginal donors.” In Western countries kidney transplants from NHBD have also been increasing. We analyzed 120 kidneys harvested from NHBD with regard to organ procurement, renal function, graft survival, and the donor factors that affected graft survival. Methods Donors were moved into the operating room after cardiac arrest. A double-balloon catheter was inserted into the abdominal aorta via laparotomy. In situ cooling by Euro-Collins solution was started at 500 mL/min. We did not performed cannulation into the femoral artery or vein prior to cardiac arrest. Results Warm ischemia time (WIT) was 18.6 minutes. Among 108 kidneys (90%) used for transplantation, 102 kidneys functioned. There were no cases of bilateral nonfunctioning kidneys. The delayed graft function (DGF) rate was 86%; however, the death-censored graft survival was 80.0% at 5 years and 62.9% at 10 years. Kidneys implanted after more than 24 hours of total ischemia time required a significantly longer period of hemodialysis. Donor risk factors that affected graft survival included WIT ≥ 20 minutes, donor age ≥ 50 years, and serum creatinine level at admission > 1.0 mg/dL. Conclusions Organ procurement without cannulation prior to cardiac arrest entailed a long WIT and a high DGF rate. However, the graft survival was good. It has been necessary to use grafts from NHBD despite the inherent risk factors. It is important to reduce kidney damage both at the organ procurement and during the posttransplant management.

PRESERVATION OF EMBRYONIC KIDNEYS FOR TRANSPLANTATION

P30 Aims: Long-term storage of embryonic kidneys is crucial for the organisation of transplantation and organ banking. To date, ex vivo preservation of rat embryonic kidneys has proved feasible for three days in ice cold University of Wisconsin solution with added growth factors. In this present study, the effects of controlled rate freezing and ice-free vitrification on metanephroi (mn) viability were investigated. Vitrification is amorphous solidification of a supercooled liquid and it is achieved by adjusting the solute composition and the cooling rate such that nucleation and growth of ice crystals is prevented. Vitrification typically employs high concentrations of cryoprotectants to prevent ice formation with the risk of cryoprotectant-induced cytotoxicity. Methods: Metanephroi were isolated from 15-day (E15) timed pregnant Lewis rats, these were then randomly divided between three groups: control (C), controlled rate freezing (CRF) and vitrification (V). CRF metanephroi were cryopreserved using a DMSO/FCS/RPMI solution and controlled rate freezing at –0.3°C/min to -40°C prior to storage at -135°C for 48 hours. Mn were re-warmed by agitation in a 37°C waterbath until thawed, cryoprotectant was them removed and mn washed prior to re-suspension in media and subsequent viability analysis. Vitrified samples were cooled to -100°C in cryoprotectant (VS55, consisting of an 8.4M mixture of DMSO, formamide, and propanediol in EuroCollins solution) using an isopentane bath, then vitrified to -120°C and stored for 48 hours at -135°C. After storage mn were thawed to -35°C in a 30% DMSO bath then the cryoprotectant was diluted out by addition of mannitol in Unisol. The supernatant was removed and mn were resuspended in media prior to viability analysis. Mn viability was analysed for cell viability via an AlamarBlue assay, histology (light microscopy, T.E.M. and cryosubstitution) and transplantation to adult Lewis recipients. Results: Following normalisation, there was statistically no difference in embryonic kidney metabolic activity of either of the cryopreserved mn group relative to the control untreated group. However, cryosubstitution demonstrated the presence of significant ice formation during controlled-rate freezing resulting in ‘scattered’ tissue and water inside the cells post-thawing. In contrast, the amount of ice was significantly reduced by vitrification. This was supported by light microscopy demonstrating significant vacuolation of the cytoplasm of control-rate frozen metanephroi following re-warming. By comparison, the re-warmed vitrified metanephroi had little cytoplasmic disruption. However, T.E.M demonstrated mitochondrial and nuclear injury at the cellular level. Transplantation of the mn is the ultimate test of viability, statistically less CRF metanephroi grew compared with control and vitrified metanephroi (p<0.05). Conclusions: There is a need for long-term storage of organs to make mn transplantation a realistic possibility. This study demonstrates that standard control-rate freezing methods are unsuitable for this purpose. Vitrification of mn using a protocol developed for blood vessels yielded more promising results but further development of the vitrification solution, exposure time and loading steps are required to reduce the observed toxicity and subsequent reduced viability.

Plasmolemmal potassium gradient does not affect lung protection by an ATP-regulated potassium channel opener.

We have previously shown that metabolic arrest induced with ATP-regulated potassium channel openers (PCOs) can improve lung preservation by adding Aprikalim (a PCO, Rhone-Poulene Roher) to modified Euro-Collins solution for pulmonary artery flush. Because the membrane hyperpolarizing effects of a PCO potentially competes with the depolarizing effects of a hyperkalemic solution, this study evaluated the effects of the potassium gradient on PCO-mediated lung protection. Twenty rabbits underwent lung protection in four groups. Group 1 underwent harvest and reperfusion as a "no ischemia" control. Groups 2, 3, and 4 underwent harvest followed by 18 hours of cold ischemic storage before reperfusion. Groups 1 and 4 received Euro Collins as the pulmonary flush at induction of ischemia. Group 2 received Euro Collins plus Aprikalim (100 microM); and group 3 received lactated Ringer's plus Aprikalim. After ischemic storage, the lungs were reperfused with autologous blood for 2 hours. Every 30 minutes, the lungs were given a 10-minute 100% fractional inspired oxygen (F(i)O(2)) challenge to measure maximal gas exchange as an indication of graft function. Repeated measures ANOVA showed Aprikalim improved graft function after 18 hours of cold ischemia (p < 0.0001). No significant differences were found when Aprikalim was used in either Euro-Collins (group 2) or lactated Ringer's (group 3) solution. The ability of the PCO Aprikalim to preserve gas exchange in a model of hypothermic pulmonary ischemia-reperfusion injury was not affected by the plasmolemmal potassium gradient. This is consistent with recent findings in myocardial protection studies that the protective effects of PCOs may be intracellular.

Alteration of cellular electrophysiologic properties in porcine pulmonary microcirculation after preservation with University of Wisconsin and Euro-Collins solutions

Background The effect of cold storage of porcine pulmonary microvessels in University of Wisconsin (UW) and Euro-Collins (EC) solutions on the cellular electrophysiologic properties remains unknown. Methods The pulmonary microarteries (PA, 381.6 ± 62.8 μm; n = 60) and microveins (PV, 360.8 ± 54.5 μm; n = 60) were incubated with Krebs (control), UW, or EC solution at 4°C for 4 hours in a myograph. The resting membrane potential and the endothelium-derived hyperpolarizing factor–mediated hyperpolarization to bradykinin (0.1 μmol/L) in the presence of inhibitors of nitric oxide and prostacyclin, N ω-nitro-l-arginine, hemoglobin, and indomethacin, in a single smooth muscle cell were directly measured. Results The resting membrane potential (−60.8 ± 1.3 mV in PA and −48.1 ± 0.7 mV in PV, n = 6) was depolarized after exposure to UW solution (to −18.4 ± 0.7 mV in PA and −13.6 ± 0.8 mV in PV; n = 8; p < 0.001). The amplitude of endothelium-derived hyperpolarizing factor–mediated hyperpolarization to bradykinin was also decreased (from 7.4 ± 0.7 mV to 2.6 ± 0.7 mV in PA and from 4.6 ± 0.5 mV to 0.9 ± 0.4 mV in PV; p < 0.001). In comparison, EC depolarized the membrane potential to a lesser extent (to −28.3 ± 0.9 mV in PA and to −21.3 ± 0.8 mV in PV; n = 8; p < 0.001) and almost abolished the hyperpolarization to bradykinin. After washout, hyperpolarization was partially restored (UW, 4.9 ± 0.7 mV in PA and 2.0 ± 0.3 mV in PV. p < 0.01; EC, 2.3 ± 0.5 mV in PA and 1.0 ± 0.3 mV in PV. p < 0.01). Conclusions Cold storage of porcine PA and PV with UW or EC solution impairs the electrophysiologic properties (hyperpolarization) related to endothelium–smooth muscle interaction. The alteration is more profound with EC than UW solution and in veins than in arteries. The findings urge further studies on lung preservation solutions.

Time-dependent morphological alterations of cold-stored small bowel in Euro-Collins and Ringer's lactate solutions

Small bowel is one of the organs that can in principle be transplanted. Optimum preservation of the organ is essential for the success of transplantation. The aim of the present study is the investigation of time-related morphological changes of rat small bowel during preservation in hypothermic Euro-Collins (EC) and Ringer's lactate (RL) solution using light microscopy and transmission electron microscopy to evaluate the integrity of intercellular complexes of mucosal epithelium, one of the tissues of the intestine that is most susceptible to ischemia. Small bowels were perfused with either EC, RL solution or physiological saline solution and were placed in the different preservation solutions at 4°C for 0, 3, 6 and 12 h. The results of our study suggest that both preservation solutions are suitable for short-term preservation of the small bowel although RL solution is more effective than EC solution. However, we conclude that further improvement of preservation solutions and/or techniques are needed to perform long-term preservation.

Are organ preservation solutions useful for the storage of isolated human islets?

Clinical islet transplantation invariably requires more than one donor per recipient. Delay between transplants could be reduced if islets were stored and transported between centers. This study assessed viability and response to glucose of isolated human islets after storage in tissue culture medium (TCM) 199 at 30°C (control), TCM 199 at 22°C (RT), University of Wisconsin solution @ 4°C (UW), or Eurocollins Solution at 4°C (EC) and compared 18 hours storage (group 1) or overnight culture followed by 4 hours storage (group 2). Insulin stimulation index (SI) (mean ± SD, n = 5), after 1 hour glucose static challenge was not significantly different ( P > .05) from islets in group 1 stored in RT 1.76 ± 1.08 or EC 1.14 ± 0.29 versus control 2.41 ± 1.13 or group 2, RT 1.73 ± 0.51, EC 2.07 ± 0.63 versus control 2.12 ± 0.58. However, SI UW was significantly lower ( P < .05) than the control in group 1 (1.19 ± 0.30) and group 2 (1.36 ± 0.34). Islet viability represented by the ATP/ADP ratio (mean ± SD, n = 5) was not significantly different after storage in RT 0.201 ± 0.159; EC 0.205 ± 0.123; or UW 0.611 ± 0.992 versus the control 0.223 ± 0.158 in group 1, and RT 0.178 ± 0.055; EC 0.137 ± 0.018; or UW 0.173 ± 0.085, compared with the control 0.199 ± 0.069 in group 2. We conclude, organ preservation solutions EC and UW do not have an advantage over TCM 199 for the storage of isolated human islets.

Effects of a bradykinin B 2 receptor antagonist on ischemia–reperfusion injury in a canine lung transplantation model

Background This study investigated the effects of a bradykinin B 2 receptor antagonist, FR173657 (FR), on ischemia–reperfusion (I/R) injury in a canine lung transplantation model. Methods Eighteen pairs of weight-matched dogs were randomly divided into 3 groups. Six pairs were assigned to the FR(D+R) group, in which FR (100 nmol/kg/h) was administered to the transplant donor continuously beginning 30 minutes before ischemia until the onset of ischemia, and FR was administered to the transplant recipient beginning 30 minutes before reperfusion and continuing for 2 hours after reperfusion. Another 6 pairs of dogs were assigned to the FR(R) group, in which FR was administered only to the recipient in the same manner as in the FR(D+R) group. The other pairs were assigned to the control group, in which vehicle alone was administered. Orthotopic left lung transplantation was performed after 12-hour cold storage in Euro-Collins solution. Fifteen minutes after reperfusion, the right pulmonary artery and the right stem bronchus were ligated. The animals were measured for 4 hours after reperfusion for left pulmonary vascular resistance (L-PVR), cardiac output (CO), arterial oxygen pressure (Pa o 2) and alveolar-arterial oxygen pressure difference (A-aD o 2). Lung specimens were harvested for measurement of the wet-to-dry lung weight ratio (WDR), histopathologic studies and polymorphonuclear neutrophil (PMN) count. Results Compared with the control group, Pa o 2, A-aD o 2, L-PVR and CO were all significantly ( p < 0.05) improved and WDR significantly ( p < 0.05) lower in both the FR(D+R) and FR(R) groups. Moreover, in the FR-treated groups, histologic tissue edema was mild, and PMN infiltration was significantly ( p < 0.05) reduced. Conclusions The bradykinin B 2 receptor antagonist, FR173657, ameliorates I/R injury in lung grafts, indicating that protection of lung grafts can be achieved by the administration of FR solely to the transplant recipient.

PVA hydrogel sheet macroencapsulation for the bioartificial pancreas

We newly developed a sheet-type macroencapsulation device entrapping rat islets from 3% polyvinyl alcohol (PVA) dissolved in Euro-Collins solution containing 10% fetal bovine serum and 5% dimethyl sulfoxide (PVA+EC) using a freezing/thawing technique. The same encapsulation technique but with 3% PVA dissolved only in double-distilled water (PVA) and a culture of free islets were served as controls. After 14-day culture in the CMRL-1066 medium, the islet recovery rate, morphological changes, insulin content, and insulin secretion were evaluated in vitro to prove the feasibility of this method of encapsulation. We also xenotransplanted the device into the peritoneal cavity of diabetic C57BL/6 mice to check its function in vivo. After 1-day culture, the islet recovery rate and insulin content in the PVA group were significantly lower than that in the PVA+EC and free islet groups. After 14-day culture, only the islets in the PVA+EC group maintained a normal morphology and effective insulin secretory response to high glucose while the response was not observed in the PVA group after 1-day culture and no longer observed in the free islets after 7-day culture. After transplantation of rat islets encapsulated in the PVA+EC device to diabetic C57BL/6 mice, nonfasting blood glucose levels showed a rapid decrease from high glucose levels of pre-transplantation, maintaining significantly lower glucose levels during the whole course of study in comparison with the sham-operated group. Our results indicated that this freezing/thawing macroencapsulation technique using 3% PVA+EC was effective for xenotransplantation of islet cells.

Rejeição aguda em transplantação renal. Análise dos factores de risco e sua influência na disfunção crónica.

The aim of this study was to study the incidence and causes of acute rejection (AR) on cadaveric renal transplants under cyclosporine A-based immunosuppression, and to investigate the AR effect on the ocurrence of chronic dysfunction (CD) and on the outcome of transplantation. We analyzed 794 renal transplants from cadaver donor between December, 1985 and December, 1999. We examined the major donor, recipient and graft-related factors and their influence in graft outcome. The diagnosis of AR was in 65% based in clinical and laboratorial findings: fever, decrease of diuresis, graft pain and/or strained and a serum creatinine increase of at least 0,4 mg/dl, and in 35% the diagnosis of AR was biopsy-proven. Statistics included univariate and multivariate analysis. Graft and patient survival rates were calculated by Kaplan-Meier method (with log-rank test). In the 794 renal transplants included in the study, 498 (63%) didn't have AR and 296 (37%) had at least one episode of AR. This overall incidence of AR of 37% is decreasing and in the last two years, 1998 and 1999 remained in 24% and 25%, respectively. The AR was associated with cadaver donor-related factors [ non traumatic cause of death (p=0,018), perfusion with Eurocollins solution (p=0,008) and cold ischemia time > 24 hours (p= 0,032)] and with recipient-related factors [age < 45 years (p=0,000) and immunosuppression with antithymocyte globulin (ATG) + azathioprine (Aza) + prednisone (Pred) + cyclosporine A (CsA) (p=0,006)]. Concerning graft-related factors acute tubular necrosis (ATN) was related with higher incidence of AR (35% of AR without ATN versus 48% of AR with ATN, p= 0,008) and this, was responsible by a significative increase of chronic dysfunction (CD) (p= 0,000) and by the worst graft function at the end of the first year (p= 0,000). Our results also showed that CD as a cause of graft loss increased substantially (23% vs 49%) in the presence of AR. The 1, 3, 5, 10 and 15-year graft survival in the patients with AR were, 94%, 85%, 74%, 50% and 38%, respectively, and in the patients without AR, 97%, 91%, 87%, 77% and 67%, respectively. These results were statistically significant ( p= 0,000). The AR didn't influence patient survival (p= 0,814). Our overall incidence of AR (37%) is decreasing reaching in the last year 25%. The AR incidence increased significantly with grafts from cadaver donors with non-traumatic cause of death, preserved with Eurocollins solution, with cold ischemia times > 24 hours and in recipients with age < 45 years and with ATG+Aza+Pred+CsA immunosuppression regimen. ATN increased the AR incidence and this was associated with a higher ocurrence of CD and a worst graft function at 1 year. The graft lost by CD duplicate in the patients with AR. Graft survival was significantly worse in the patients with AR. The AR did not adversely affect patient survival.

Inhibition of c-Jun NH2-Terminal Kinase Activity Improves Ischemia/Reperfusion Injury in Rat Lungs

Although c-Jun NH(2)-terminal kinase (JNK) has been implicated in the pathogenesis of transplantation-induced ischemia/reperfusion (I/R) injury in various organs, its significance in lung transplantation has not been conclusively elucidated. We therefore attempted to measure the transitional changes in JNK and AP-1 activities in I/R-injured lungs. Subsequently, we assessed the effects of JNK inhibition by the three agents including SP600125 on the degree of lung injury assessed by means of various biological markers in bronchoalveolar lavage fluid and histological examination including detection of apoptosis. In addition, we evaluated the changes in p38, extracellular signal-regulated kinase, and NF-kappaB-DNA binding activity. I/R injury was established in the isolated rat lung preserved in modified Euro-Collins solution at 4 degrees C for 4 h followed by reperfusion at 37 degrees C for 3 h. We found that AP-1 was transiently activated during ischemia but showed sustained activation during reperfusion, leading to significant lung injury and apoptosis. The change in AP-1 was generally in parallel with that of JNK, which was activated in epithelial cells (bronchial and alveolar), alveolar macrophages, and smooth muscle cells (bronchial and vascular) on immunohistochemical examination. The change in NF-kappaB qualitatively differed from that of AP-1. Protein leakage, release of lactate dehydrogenase and TNF-alpha into bronchoalveolar lavage fluid, and lung injury were improved, and apoptosis was suppressed by JNK inhibition. In conclusion, JNK plays a pivotal role in mediating lung injury caused by I/R. Therefore, inhibition of JNK activity has potential as an effective therapeutic strategy for preventing I/R injury during lung transplantation.

Bronchoscopic surfactant administration preserves gas exchange and pulmonary compliance after single lung transplantation in dogs

Background Surfactant abnormalities have been implicated in reperfusion injury and respiratory failure in lung transplantation. Methods We investigated the efficacy of bronchoscopic administration of a bovine natural lung surfactant extract (Alveofact) to improve gas exchange and lung mechanics after heterologous left lung transplantation in foxhounds (+4°C ischemia for 24 hours, conservation with Euro-Collins solution). Animals received either no surfactant therapy (untreated controls, n = 6) or 50 mg/kg body weight (prior to explantation, only graft) and 200 mg/kg body weight Alveofact (immediately after reperfusion, both lungs, n = 6). After lung transplantation, separate but synchronized ventilation of each lung was performed in a volume-controlled, pressure-limited mode for 12 hours, with the animals prone. Small catheters were inserted into the pulmonary veins of both the graft and the recipient's native lung for separate blood gas analysis. In the control group, marked protein leakage, influx of neutrophils into the alveolar space, and pulmonary edema formation (extravascular lung water; wet/dry ratio) were encountered in the transplanted lung but only to a very minor extent in the recipient's native lung. Results Lung compliance values and arterial oxygenation progressively deteriorated in the transplanted but not in the native lungs. Pulmonary hemodynamics did not change significantly. Surfactant administration did not significantly influence the development of reperfusion edema, protein leakage, and neutrophil influx into the grafts. However, surfactant restored the surface activity and the gas exchange (Pa o 2/F io 2 of 201.2 ± 20.2 mm Hg vs 119.8 ± 21.7 mm Hg in controls; P < .05) in the transplanted lungs, and compliance was markedly improved in the surfactant-treated animals (18.8 ± 1.8 mL/mbar vs 11.5 ± 1.6 mL/mbar in the controls; P < .05). Conclusion Bronchoscopic surfactant administration does not prevent leukocyte influx or vascular leakage but does protect against respiratory failure and improves lung mechanics in single lung transplantation in dogs.

Surfactant abnormalities after single lung transplantation in dogs: impact of bronchoscopic surfactant administration

Objective Disturbances of the alveolar surfactant system have been implicated in the pathogenesis of reperfusion injury. The aim of this study was to evaluate the influence of exogenous surfactant administration on surfactant properties in a model of single lung transplantation. Methods We performed heterologous, left lung transplantation (+4°C ischemia; 24 hours, Euro-Collins solution) in 6 foxhounds (untreated) and in 6 animals that received calf lung surfactant extract (Alveofact) prior to explantation (only donor lung; 50 mg/kg body weight) and immediately after onset of reperfusion (both lungs, 200 mg/kg body weight). Separate but synchronized ventilation of each lung was performed, in a volume-controlled, pressure-limited mode, with animals in prone position. Bronchoalveolar lavage fluids were collected in pretransplantation lungs (control), after 24 hours of ischemia prior to transplantation (0 hours) and 6 and 12 hours after reperfusion in both the grafts and the recipient native lungs. Results Ischemic storage per se did not provoke any changes of the surfactant system; however, severe alterations occurred within 6 hours of reperfusion, resulting in a severe loss of surface activity, including a decrease in the percentage of the large surfactant aggregate fraction, reduction of the surfactant apoproteins SP-B and SP-C, the dipalmitoyl molecular species of phosphatidylcholine and phosphatidylglycerol within the large surfactant aggregate fraction. These abnormalities were restricted to the graft, with virtually normal surfactant function and composition being found in the recipient native lung. Surfactant administration fully normalized the biochemical and largely improved the biophysical surfactant properties, alongside maintenance of lung gas exchange properties. Conclusions Severe surfactant abnormalities occur exclusively in the graft when performing separate, synchronized ventilation of each lung to attenuate ventilator-induced lung injury. Bronchoscopic surfactant administration provides protection against these abnormalities and may be a therapeutic strategy in lung transplantation.

Trasplante renal pediatrico con injertos de donantes infantiles de corta edad (. 6 anos). complicaciones y evolucion funcional

Pediatric renal transplantation from young children donors (aged ≤ 6 years).complications and functional evolution GoalsTo evaluate functional results and complications in a group of pediatric patients who received kidney transplants from donors aged 6 years or less. Material & methodsThirty-four consecutive renal transplants were reviewed. Of 34 patients, 19 (55.8%) received kidneys from donors aged below 3 years (Group A) and 15 (44.2%) from donors aged 3-6 years (Group B). Long-term functional patient and graft survival results along with complications were compared in both groups. ResultsSeven patients in group A (36.7%) had vascular complications, in comparison to 3 in group B (20%, p 0.05). Kidneys preserved in Eurocollins (EC) solution had more vascular complications (35.3%) in comparison to University of Wisconsin (UW) solution kidneys (23.5%, p 0.05). Ten-year graft survival rate in group A was significantly lower in comparison to group B (35,5% vs 58,6%, p 0,05). In relation to preservation solution type used, 10-year graft survival rate was also significantly better in UW group (26.8% vs 63%, p 0.001). ConclusionThe exclusion of donors younger than 3 years and the utilization of optimal preservation solutions produces a decrease in figures of vascular complications and a significant improvement in graft survival rates.

Pravastatin decreases cold ischemia–but not alloantigen-dependent transplant arteriosclerosis

Background Alloantigen mismatch and cold ischemia have been shown to induce transplant arteriosclerosis. Pravastatin (PR) decreases arteriosclerosis probably related to an immunosuppressive effect. Statins posses other nonimmune properties that may be beneficial to transplantation. We studied the effect of PR on cold ischemia and alloantigen-induced transplant arteriosclerosis in syngeneic (SYN) and allogeneic (ALLO) aortic transplantation models. Methods Lewis rats served as the donors and recipients for SYN transplants and Brown Norway rats were donors for ALLO transplants. Aortic segments that had been preserved at 4°C in Euro-Collins solution for 0 or 24 hours were transplanted to the infrarenal aorta of the recipients PR (10 mg/kg/d) was administered for 12 weeks prior to morphometric studies. Areas of intimal thickness and its relation to total vessel area were calculated. Lipid levels were measured at 12 weeks. Results Aorta rings preserved for 24 hours showed marked intimal thickening compared to controls (SYN, CI 0 hours = 21.5% ± 16.5% vs SYN, CI 24 hours = 50.7 ± 9.5%, P < .05). PR significantly decreased thickening (SYN, CI 24 hours + PR = 41.7 ± 12.2 ( P < .05) vs SYN, CI 0 hours on SYN, CI 24 hours). There was a nonsignificant decrease in thickening among ALLO transplants treated with PR (ALLO = 31.4 ± 15.9 vs ALLO + PR = 23.8 ± 18.8; P > .05). PR had no effect on lipid levels. PR decreases cold ischemia induced transplant arteriosclerosis in this syngeneic aortic transplant model, but does not affect an alloantigen-mediated process. The beneficial effect of PR is not related to its lipid-lowering properties but probably to a nonimmune effect.