Assessment of the Learning Process After Introducing a New Liver Transplantation Procedure in a Center With Extensive Experience in Kidney Transplantation.

BACKGROUND: Liver transplantation (LTx) is one of the most complex transplant procedures. The aim of the present study was to determine whether the learning process can be observed after the introduction of LTx in a center with extensive previous experience in renal transplantation. METHODS: This retrospective analysis included 264 primary LTx procedures performed with the piggyback technique (2005-2016). The procedures were divided into 4 equal groups. The characteristics of the recipients, data related to the surgery, and the postoperative course and complications were analyzed. RESULTS: We observed a significant reduction in surgical time and in the anhepatic phase duration between Group 1 and the other groups (median surgical time was 455 minutes vs 415 minutes, 410 minutes and 387 minutes, respectively, P < .05; median anhepatic phase duration was 75 min vs 60 min, 62 min, 60 min, respectively, P < .05). There was a decrease in the number of transfused blood units (median in Group 1 of 6 packs vs 3 packs in Group 4, P < .05) and a decrease in blood recovered from the operating field using the Cell Saver system (median in Group 1 of 1570 mL vs 1057 mL, 1123 mL, and 1045 mL, respectively, P < .05). A significant reduction in the number of hemorrhages was found (1.5% in Group 4 vs 13.6%, 10.6%, and 7.6% in the other groups P < .05). The remaining studied parameters were not statistically significant. CONCLUSIONS: Extensive previous transplantation experience affected the lack of typical features of the learning process.

Influence of Experience Acquired by a Liver Transplantation Center on Extension of Donor Acceptance Criteria.

BACKGROUND: Liver transplantation (LTx) is the only effective treatment for end-stage liver failure. Due to the ongoing lack of organs available for transplantation, there is a tendency to extend liver donor selection criteria. The aim of the study was to determine whether extension of donor acceptance criteria with increasing experience in LTx occurred at our transplant center. METHODS: This retrospective analysis included 288 donors harvested between 2005 and 2016. The donors were divided chronologically into 4 equally sized groups. They were assessed in subsequent groups according to sex, age, height, body mass index (BMI), cause of death, amount of days spent in the intensive care unit, number of episodes of cardiac arrest before organ removal, and results of laboratory and virologic tests. RESULTS: A statistically significant increase in the age of accepted donors was observed between group 2 and group 4 (median 40 vs 45 years, P < .05). There was a significant increase in the acceptance of anti-HBc-positive donors (0% in group 1 vs 7% in group 4). The remaining parameters did not show statistically significant differences. CONCLUSION: Experience acquired by our transplant center during the period of analysis did not lead to extension of liver donor acceptance criteria. Statistically significant differences for liver donor age and virologic profile (anti-HBc) between groups were observed; however, overall analysis did not confirm a clear tendency to extend liver donor acceptance criteria at this center.

Comparison of Vascular Endothelial Growth Factor Concentration in Porcine Kidneys Removed From Living Donors After Cardiac and Brain Death.

The aim of this paper was to describe the differences in vascular endothelial growth factor (VEGF) concentration in porcine kidneys removed from living donors (group I), donors after prior induction of brain death by brain herniation (group II), and donors after cardiopulmonary arrest (group III). The groups consisted of 6 animals which underwent dual renal removal procedures; kidneys were rinsed, stored for 24 hours at 4°C and rinsed again. Renal specimens (4g) were collected before and after perfusion (time 0 and 1), after 12 hours (time 2), and after reperfusion (time 3). A Western blot was used to evaluate VEGF concentration in collected tissues homogenates. Additionally, the levels of VEGF, interleukin 1ß, tumor necrosis factor α, and endothelial nitric oxide synthase (eNOS) were detected with enzyme-linked immunosorbent assays. Directly after the removal procedure, no significant differences in VEGF levels (IOD) were observed depending on the donor (moderate levels were observed in all groups: 1.51 in group I, 1.48 in group II, and 1.35 in group III). As a consequence of perfusion and 12 hours of storage, a stable concentration in groups I and III was observed with a gradual increase of VEGF levels in group II (1.23, 2.08, and 1.67 in the respective groups at time 1; 1.49, 2.12, and 1.63 in the respective groups at time 2). After the following 12 hours, a statistically significant (P < .05) higher level of VEGF was observed in group II (2.34) in comparison to groups I and III (1.58 and 1.81, respectively). In group I, a correlation between VEGF concentration and IL-1ß was observed, while in group II there was correlation between VEGF and eNOS levels.

Results of Transplantation of Kidneys Procured From Donors After Brain Death Aged 60 Years and Older.

OBJECTIVE: To analyze results of transplantation of kidneys procured from donors after brain death aged 60 years and older (hereafter denoted by "≥60") compared to kidneys procured from donors after brain death aged 40-59 years (hereafter denoted by "40-59") in medium-term follow-up period, and to assess factors that affect recipient and kidney graft survival. MATERIAL AND METHODS: 92 transplant recipients of kidneys procured from donors after brain death ≥60 were enrolled into the study. The control group were 363 recipients of kidneys procured from donors after brain death 40-59. RESULTS: Mean values of serum creatinine were higher in recipients of kidneys procured from donors after brain death ≥60 compared to control after 3 years: 168.2 ± 57.5 (n = 59) vs 147.9 ± 65.7 (n = 294), P < .05; and after 5 years: 196.2 ± 95.3 (n = 38) vs 157.3 ± 80.0 µmol/L (n = 211), P < .01. Restricted mean recipient survival time was 56.4 (95% confidence interval: 55.0-57.8) and 52.0 (48.0-56.1) months, P < .05; and kidney graft survival time was 51.6 (49.6-53.5) and 43.9 (39.0-48.9) months, P < .01 in recipients who received kidneys from donors after brain death 40-59 and from donors after brain death ≥60 respectively. In Cox regression, donor death due to cardiovascular disease proved to be the factor increasing risk of kidney graft loss (hazard ratio 1.553, P < .001). CONCLUSIONS: The survival and function of kidneys procured from donors after brain death ≥60 at medium-term follow-up remain worse compared to kidneys procured from donors after brain death 40-59, and the donor dependent risk factor of kidney graft loss is cardiovascular disease, which caused donor death.

Effect of the Second Warm Ischemia Time and Its Components on Early and Long-term Kidney Graft Function.

BACKGROUND: During kidney transplantation, the total time of organ ischemia consists of first warm ischemia time (WIT1), cold ischemia time (CIT), and a second WIT (WIT2). Rising graft temperature during WIT2, which comprises the creation of vascular anastomoses, increases oxygen demand and tissue damage, especially in the kidney tubular cells. The aim of this study was to analyze the influence of WIT2 on early and long-term kidney graft function. METHODS: We performed a retrospective analysis of 554 consecutive adult recipients, who received their first kidney graft from a deceased donor between 2003 and 2013. RESULTS: Mean WIT2 was 25.2 min. Donors' sex, age, presence of hypertension, body mass index (BMI), and the cause of brain death showed no effect on WIT2. Weak positive correlations were found between the duration of WIT2 and both recipients' age (r = 0.11; P < .01) and BMI (r = 0.14; P < .01). Multivariate regression analysis confirmed the independent influence of age (ß = 0.107 [95% confidence interval, 0.017 to 0.197] per year; P = .02) but not BMI (P = .09). WIT2 influenced early graft function and was significantly longer in patients with primary graft nonfunction than in other recipients (35.3 vs 24.9 min; P < .01). According to receiver-operating characteristic curve analysis, a WIT2 value >26 min was predictive of primary graft nonfunction, with 64% specificity and 58% sensitivity. No correlations were found between WIT2 and estimated glomerular filtration rate in the long-term follow-up period. CONCLUSIONS: This study found that WIT2 may significantly influence the early graft function. We also found that the creation time of vascular anastomoses does not affect the long-term kidney graft excretory function.

Early and Long-Term Outcomes of Kidney Grafts Procured From Multiple-Organ Donors and Kidney-Only Donors.

BACKGROUND: The deceased-donor kidney pool consists of 2 different populations: multiple-organ donors (MOD) and kidney donors alone (KDA). In MOD, more complicated procedure and lowest priority for kidney procurement may affect graft survival. On the other hand, poor donor status and higher comorbidity are more frequent in KDA transplants. The aim of this study was to provide detailed characteristics of the 2 groups of kidney donors (KDA vs MOD) in our center and to analyze the potential influence of the donor type on the early and long-term kidney graft function and recipient outcome. METHODS: We performed a retrospective analysis of 729 first cadaveric kidney transplant recipients: 499 of them received the organ from MOD, 230 from KDA. RESULTS: The frequency of delayed graft function (DGF) was higher in KDA than in MOD transplants (38.7 vs 25.1%; P < .001). Multivariate logistic regression analysis revealed that donor age, KDA, and early acute rejection independently increased the risk of DGF occurrence, whereas recipient age and cold ischemia time increased the risk of primary graft nonfunction. Kidney excretory function was significantly worse in KDA up to 10 years after transplantation. There were no differences in kidney graft and patient survivals, frequency of proteinuria, acute rejection, and cytomegalovirus episodes, and post-transplantation diabetes. CONCLUSIONS: (1) The use of a kidney from KDA negatively affects early and late kidney graft function compared with MOD. (2) The long-term kidney graft and patient survivals are not affected by the type of organ procurement.

Donation and Transplantation of Kidneys Harvested From Deceased Donors Over the Age of 60 Years in the Upper Silesia Region.

BACKGROUND: Population aging and shortage of organs for transplantation result in increasing numbers of kidneys retrieved from elderly donors. The aim of this study was to analyze donation of kidneys from donors after brain death (DBD) over the age of 60 years (≥60), comorbidities that affect decisions on retrieval, and early results of kidney transplantation. METHODS: Ninety-six potential DBD ≥60 and 309 aged 40-59 years (40-59) reported in Upper Silesia, Poland, from 2004 to 2013 were enrolled in the study. RESULTS: DBD >60 presented a higher rate of coexisting hypertension (53% vs 34%), limb ischemia (10% vs 1%), and past stroke (6% vs 1%) compared with DBD 40-59 (P < .05), but no differences were observed in serum creatinine concentration (85 vs 84 µmol/L), coexisting coronary disease (14% vs 6%), or diabetes (10% vs 4%). The decision of withdrawal from retrieval was more frequent in DBD ≥60 (16% vs 7%; P < .05). Twelve months after kidney transplantation, serum creatinine concentration was higher in recipients of kidneys from DBD ≥60 compared with DBD 40-59 (169 vs 138 µmol/L; P < .001). The survivals of recipients (93% vs 95%) and kidney grafts (90% vs 93%) as well as rates of proteinuria >1.0 g/24 h (6% vs 2%) did not differ between the groups. CONCLUSIONS: A higher rate of comorbidities in potential kidney DBD ≥60 results in a lower retrieval rate in these donors. The function of kidneys harvested from DBD ≥60 12 months after transplantation is worse than those from DBD 40-59, but still acceptable.

Past Cardiovascular Episodes Deteriorate Quality of Life of Patients With Type 1 Diabetes and End-stage Kidney Disease After Kidney or Simultaneous Pancreas and Kidney Transplantation.

BACKGROUND: The beneficial influence of kidney (KTx) or simultaneous pancreas and kidney transplantation (SPK) on quality of life (QOL) in patients with end-stage kidney disease caused by type 1 diabetes mellitus was confirmed in many studies. The aim of this study was to identify factors that influence QOL of patients in long-term follow-up after SPK or KTx. METHODS: Twenty-seven SPK and 26 KTx patients with good function of transplanted organs at least 1 year after transplantation were enrolled into the analysis. To estimate QOL of the recipients the Kidney Disease and Quality of Life Short Form was applied. RESULTS: Within the whole analyzed group, the necessity of exogenous insulin administration correlated (P < .05) with symptom/problem list (γ = -0.35), effects of kidney disease (-0.38), cognitive function (-0.47), sleep (-0.42), overall health (-0.47), physical functioning (-0.61), role-physical (-0.32), pain (-0.50), general health (-0.32), emotional well-being (-0.31), role-emotional (-0.36), social function (-0.33), energy/fatigue (-0.44), and the SF-12 physical composite (-0.44). History of cardiovascular episode correlated (P < .05) with symptom/problem list (γ = -0.59), effects of kidney disease (-0.46), burden of kidney disease (-0.56), sleep (-0.54), social support (-0.51), physical functioning (-0.55), role-physical (-0.70), pain (-0.60), general health (-0.57), emotional well-being (-0.45), role-emotional (-0.95), social function (-0.58), energy/fatigue (-0.59), SF-12 physical composite (-0.45), and SF-12 mental composite (-0.83). CONCLUSIONS: Exogenous insulin administration and history of cardiovascular episode are the most important factors influencing QOL in patients after SPK or KTx, particularly worsening its physical components.

Long-Term Outcomes of Kidney and Simultaneous Pancreas-Kidney Transplantation in Recipients With Type 1 Diabetes Mellitus: Silesian Experience.

BACKGROUND: Kidney transplantation (KTx) markedly reduces mortality in patients with end-stage kidney disease (ESKD) caused by type 1 diabetes mellitus (T1DM). The outstanding issue is whether transplantation should be limited only to KTx, with further insulinotherapy, or combined with pancreas transplantation in patients with ESKD/T1DM. The goal of this study was to compare the results of simultaneous pancreas-kidney transplantation (SPKTx) and deceased donor KTx and to identify factors affecting patient and kidney graft survival in patients with ESKD/T1DM. METHODS: Eighty-seven deceased donor KTx and 66 SPKTx operated on in the Silesia region of Poland between 1998 and 2013 were included in the retrospective analysis. RESULTS: During the mean 6.7 ± 3.6 years of follow-up, fewer cardiovascular episodes were observed in SPKTx recipients than in KTx recipients (1.5% vs 12.6%; P < .05). Five-year patient survival (80.7% in SPKTx vs 77.5% in KTx) and kidney graft survival (66.1% in SPKTx vs 70.4% in KTx) did not differ between study groups. There were no differences in patient survival (log-rank test, P = .99) or kidney graft survival (P = .99) based on Kaplan-Meier curves. Multivariable Cox proportional hazard analysis failed to identify factors explaining patient and kidney graft survival. Five-year pancreas graft survival was 58.9%. SPKTx recipients had significantly higher estimated glomerular filtration rates during the 7-year posttransplant period and less frequently developed proteinuria (6.1% vs 23%; P < .01). CONCLUSIONS: Pancreas transplantation reduced cardiovascular risk and prevented the development of proteinuria but did not improve patient and kidney graft survival in recipients with T1DM in the 7-year follow-up period.

Caspase-3 Expression and ALT, AST, and GGT Activity After 24 Hours of Porcine Liver Cold Storage, Depending on the Type of Transgenesis.

BACKGROUND: Because of an insufficient number of human organs for transplantation, xenotransplantation may become an effective alternative. We aimed to analyze if the type of transgenesis has an influence on the hepatic caspase-3 expression, the enzyme that executes apoptosis as well as ALT, AST, and GGT activity after 24 hours of cold storage. METHODS: The experiment was carried out on the 24 livers of Polish White Landrace pigs carrying human α1,2-fucosyltransferase and/or α-galactosidase (GAL) genes and livers without this genetic modification (control). Livers were perfused, stored for 24 hours in solution, and subsequently re-flushed. Hepatic concentration of the caspase-3 protein and its mRNA expression were measured just after the animal was killed as well as after 30 minutes of perfusion and after 24 hours of cold storage followed by 30 minutes of reperfusion. Caspase-3 mRNA level was detected with the RT-PCR method. Protein concentration (capsase-3 active and inactive) was assessed with the Western blotting technique. Kinetic methods were applied for the analysis of the ALT, AST, and GGT activity. RESULTS: The highest increase of the ALT activity after cold storage was observed in the group with GAL transgenesis, whereas the GGT activity was highest in the unmodified livers. There was no difference in the caspase-3 expression and AST activity after cold storage as compared with the respective initial results (P = .57 and P = .97, respectively). CONCLUSIONS: It appears that transgenesis does not aggravate ischemic injury of the liver.

Endothelial Nitric Oxide Synthase and Endothelin-1 Expression in the Early Post-Porcine Kidney Autotransplantation Period.

BACKGROUND: The aim of this study was the assessment of endothelial nitric oxide synthase (eNOS) and endothelin-1 (EDN-1) expression in porcine kidneys on the 14th and 30th days after the autotransplantation procedure. METHODS: The research was conducted on 12 animals that underwent a left renal transplantation procedure with further standardized rinsing and 24-hour storage in 4°C; subsequently, the kidneys were implanted in the right retroperitoneal space after right-sided nephrectomy. Removed kidneys were examined (group 0). Six randomly chosen animals (group 1) were under observation for 14 days and 6 others (group 2) for 30 days. RESULTS: After these observation periods, euthanasia was performed on the animals and 4-g samples were collected from the renal cortex and medulla. The Western blot technique was used to detect the eNOS and EDN-1 expression at the protein level. The obtained results are presented as absolute values of integrated optical density. Stable graft function was observed in all animals from the 2nd day after the procedure. eNOS in group 1 reached the mean value of 1.064 and was statistically significantly lower than in group 2 (2.085) or in the control group 0 (3.318). In the case of EDN-1 expression on 14th day after transplantation, the medium level was reported (0.248), which was similar to group 0 (0.216), whereas group 2 presented values 2 times higher (0.743). CONCLUSIONS: A lowered eNOS level in the organ was observed on the 14th day after autotransplantation of a pig kidney; further enzyme normalization is associated with increased EDN-1 expression.

Concentration of Tumor Necrosis Factor-α and Interleukin-1ß in Isolated Porcine Liver Depending on Type of Transgenesis.

BACKGROUND: Transgenic animals may serve as organ donors in human organ transplantation. However, the number of the studies addressing all doubts related to this issue is currently insufficient for the clinical application of this approach. The aim of this study was to analyze the hepatic tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) synthesis during a 24-hour cold preservation of the transgenic pig liver, depending on the type of transgenesis. MATERIALS AND METHODS: The study was carried out on wild-type and transgenic pig livers with transferred human α1,2-fucosyltransferase (FUT) and/or α-galactosidase (GAL) gene (four groups; n = 6). Harvested livers were perfused for 30 minutes and stored for 24 hours in Biolasol (Biochefa) solution at 4°C with a subsequent 30-minute reperfusion (reflush). TNF-α and IL-1ß concentrations were analyzed with an enzyme-linked immunosorbent assay. Perfusates were collected during the initial perfusion as well as after 24 hours of preservation and during the reperfusion. Tissue samples were harvested just after animal sacrifice, and after organ perfusion and reperfusion. RESULTS: A decrease in TNF-α concentration in homogenates was noted after both perfusion and reperfusion in all experimental groups. In contrast, there was a significant decrease in IL-1ß concentration in the group with combined human FUT and GAL transgenes. Concurrently, increases in TNF-α and IL-1ß concentrations were observed in the reperfusion perfusates in all groups. CONCLUSION: This study shows that IL-1ß is synthesized in the ischemic livers of the transgenic animals with both human α1,2-fucosyltransferase and α-galactosidase transgenes. Further analysis is required to determine the importance of this observation.

Biolasol: novel perfusion and preservation solution for kidneys.

INTRODUCTION: Biolasol solution (Pharmaceutical Research and Production Plant "Biochefa," Sosnowiec, Poland) is a novel extracellular perfusion and ex vivo hypothermic kidney preservation solution. It ensures maintenance of homeostasis, reduces tissue edema, has low viscosity, and allows the graft to preserve structural and functional integrity. It minimizes ischemia-reperfusion damage. METHODS: Perfundates from control and transplanted kidneys flushed with Biolasol or ViaSpan solutions (Arkas, Warszawa, Poland) were analyzed. Parameters of serum and urine collected from 12 pigs after auto-transplantation were also analyzed. Renal medulla was investigated for structural alterations by analyzing hematoxylin and eosin-stained slides. The mean survival time of pigs after the auto-transplantation procedure was the measure for the novel Biolasol solution effectiveness. RESULTS: We observed a statistically significant decrease in marker enzyme levels alanine transaminase, aspartate transaminase, lactic dehydrogenase, and ions (Na and K) in pigs with grafts flushed with Biolasol. Histopathologic examination revealed that the renal cortex structure was not damaged after the use of Biolasol solution. CONCLUSION: Biolasol solution protects kidneys against ischemia damage and does not differ significantly from the "golden standard" ViaSpan solution.

Comparison of endothelial nitric oxide synthase and endothelin-1 levels in kidneys removed from living pigs after cardiac arrest and brain death.

OBJECTIVES: The aim of this paper was to describe differences between levels of endothelial nitric oxide synthase (NOS-3) and endothelin-1 (ET-1) in swine kidneys removed from living donors (group I) and after inducing brain death by brain herniation (group II) and cardiac arrest (group III). METHODS: Each group consisted of 3 animals who underwent dual renal removal procedure; kidneys were further rinsed according to standardized procedure with Biolasol perfusion liquid, stored for 24 hours (4°C), and rinsed again. Renal specimens of 4 g mass, including renal cortex and medulla, were collected before and after perfusion (times 0 and 1), after 12 hours (time 2), and after reperfusion (time 3). Enzyme-linked immunosorbent assay was used to describe levels of NOS-3 and ET-1 in collected tissues homogenates. Mann-Whitney U test was used to compare results in groups in relation to total protein content (ng/mg), and the correlation between the 2 substances was measured with the use of Spearman rho. RESULTS: Group I presented low and stable levels of NOS-3 in all time intervals (averages, 0.73, 0.99, 0.52, and 0.89, respectively). Level sof ET-1 were similar (0.87, 0.63, 0.69, and 0.86, respectively), and significant correlation between levels of the 2 substances was observed. Increased levels of NOS-3 (1.89 and 1.86) and ET-1 (1.38 and 1.49) were observed directly after removal in groups II and III and further maintained during organ storage. No correlation in group I was observed, and after perfusion significantly lower level of NOS-3 was observed in kidneys removed after brain death in relation to group III (1.77 vs 2.60). CONCLUSIONS: The lowest and stable levels of NOS-3 and ET1 during storage were observed in kidneys removed from living donors. Levels of analyzed substances in this group showed correlation in subsequent time intervals.

Toll-like receptors 2 and 4 in pigs' kidneys early after autotransplantation procedure.

OBJECTIVES: The aim of this paper was to evaluate mRNA expression of Toll-like receptors 2 (TLR2) and 4 (TLR4) and the adaptor protein myeloid differentiation primary-response protein 88 (MyD88) in pigs' kidneys 14 and 30 days after autotransplantation. METHODS: The research was conducted on 12 animals that underwent left renal transplantation procedure with further standardized rinsing with Biolasol solution and 24 hours' storage in 4°C; subsequently the kidneys were implanted in the right retroperitoneal space after right-side nephrectomy. Six randomly chosen animals (group I) were under observation for 14 days, the other 6 (group II) for 30 days. After these observation periods, the animals were killed and 4-g samples were collected from the renal cortex and medulla. RESULTS: Expression of mRNA in homogenates of collected samples were determined with the use of reverse-transcription polymerase chain reaction analysis. Obtained results in both groups, presented in relation to GAPDH, were compared with the use of Mann-Whitney U test. Stable graft function was observed in all animals from the 2nd day after the procedure. TLR2 in group I reached the mean value of 3.64 and was statistically significantly higher than in group II (2.19). Inverse proportion was observed in case of mRNA for TLR4: group II presented 2 times higher value than group I (0.25 vs 0.11). Similarly, significant difference was observed in MyD88 (group I, 0.067; group II, 0.45). CONCLUSIONS: At 14 days after autotransplantation of a pig kidney, mRNA expression for TLR2 is dominant; later, expression increases for TLR4 and MyD88.

Cytochrome P450 3 A expression in pigs livers after 24-hour preservation in Biolasol solution depending on the type of transgenesis.

BACKGROUND: An insufficient number of organs for transplantation shows the need for the development of new technologies. Xenotransplantation might be the answer. OBJECTIVE: To determine if the type of transgenesis influences the level of CYP3A4, which takes an active part in xenobiotics metabolism in livers after 24-hour storage, depending on the kind of solution used for preservation. MATERIALS AND METHODS: The experiment was carried out on 30 livers of Polish White Landrace divided into 5 groups depending on transgene type. The following human genes were transferred: α1,2-fucosyltransferase (groups I and II), α-galactosidase (III), combined α1,2-fucosyltransferase/α-galactosidase transgene (IV), and livers without modification (V). The livers were perfused and subsequently stored for 24 hours in Ringer's solution (group I) or Biolasol solution (II-V). Reperfusion/reflush was performed. CYP3A29 isomer concentration was analyzed in liver specimens collected twice: 30 minutes after perfusion and 30 minutes after reperfusion/reflush. Expression of mRNA CYP3A29 was marked using RT-PCR analysis and of protein CYP3A29 using Western blotting technique. RESULTS: The most significant decrease in protein CYP3A29 expression after 24-hour preservation was observed in group I (55.88% decrease), while the least significant was observed in group IV (10.44% decrease). mRNA expression evaluation was similar: the most significant decrease was observed in group I (87.8% decrease) and the least significant in group III (4.6% decrease). CONCLUSION: α1,2-Fcosyltransferase transgene seems to influence mRNA and protein CYP3A expression in case of liver grafting and preservation for transplantation. CYP3A expression was also influenced by the kind of preservation solution used.

Interleukin-6 concentration in the transgenic pig's liver preserved for 24 hours in Biolasol solution.

INTRODUCTION: Increasing the human lifespan contributes to a higher number of patients with end-stage organ failure, which in turn stimulates the search for alternative sources. Xenotransplantation seems to be a promising approach in this respect. OBJECTIVE: Analysis of changes in interleukin (IL)-6 concentration during 24-hour preservation of transgenic swine livers, depending on the kind of transgenesis and preservation solution used. MATERIALS AND METHODS: The experiment was carried out in swine livers with transferred human genes that were divided into 5 groups. The following human genes were transferred: α1,2-fucosyltransferase (group I and II), α-galactosidase (III), combined α1,2-fucosyltransferase/α-galactosidase transgene (IV), and livers without modification (V). The livers were perfused and subsequently stored for 24 hours in Ringer's (group I) or Biolasol solutions (II-V). Reflush was then performed. IL-6 concentration was analyzed in the solution samples collected at the beginning and end of perfusion, and after 24 hours of preservation. ELISA was used to evaluate IL-6 concentration. RESULTS: In liver homogenates from group I, IL-6 concentration after 24 hours of preservation increased by 8.24% compared to the levels observed after perfusion, whereas in the other groups IL-6 concentration decreased. The most significant decrease, 49.51%, was observed in group II; the least significant in group IV, 10.72%. In case of supernatants, a statistically significant increase of AUC0-30min level in relation to perfusion was observed in every group after 24-hour preservation and reperfusion. The highest values of AUC0-30min were observed in group I (α1,2-fucosyltransferase, Ringer's solution). CONCLUSION: The study indicates the hepatoprotective action of Biolasol solution.

Splenectomy attenuates the course of kidney ischemia-reperfusion injury in rats.

INTRODUCTION: Renal ischemia-reperfusion injury (IRI) initiates inflammatory response with synthesis of free oxygen radicals, chemokines, and cytokines which attract neutrophils and monocytes, which then differentiate into macrophages and dendritic cells, activating adaptive immune response. The spleen is the main source of both monocytes and lymphocytes. The aim of this study was to assess whether splenectomy performed before or upon IRI affects post-ischemic and long-term renal function. METHODS: Two weeks after right nephrectomy, the left kidney pedicle was clamped for 45 minutes in 24 rats. After the clip insertion, the spleen was removed in 12 animals and the remaining 12 rats underwent sham splenectomy. In the second experiment, splenectomy (n = 9) or sham procedure (n = 9) was performed simultaneously with right nephrectomy, 2 weeks before left kidney ischemia. The excretory function of the kidney was evaluated 48 hours and 7 days after ischemia. In the experimental model of chronic renal failure, 14 days before right nephrectomy, the prolonged 90-minute ischemia was induced in 32 rats with simultaneous splenectomy (n = 16) or sham procedure (n = 16). In long-term observation, the renal function and mortality rate was evaluated. RESULTS: Kidney function preservation was superior in rats that underwent splenectomy together with renal ischemia when compared to controls. This was further expressed with a 2 times lower mortality rate in splenectomized animals in 6 months observation after prolonged renal ischemia. Renoprotective effect was not observed when splenectomy was performed 2 weeks before IRI. CONCLUSIONS: The results suggest a detrimental influence of the spleen on the development of renal IRI.

The effect of HTK solution modification by addition of thyrotropin and corticotropin on biochemical indices reflecting ischemic damage to porcine kidney.

INTRODUCTION: The aim of our study was to evaluate the impact of perfusion with HTK (histidine-tryptophan-ketoglutarate, Custodiol®, Dr. Franz Kohler Chemie, Germany) solution, modified by the addition of porcine thyroid-stimulating hormone (TSH) and corticotropin (ACTH), on selected biochemical parameters of porcine renal damage within 24 and 48 hours after the onset of cold ischemia time. METHODS: Each study group consisted of 10 adult pigs. During harvesting the kidneys were rinsed with Ringer solution (group 1), HTK (group 2), HTK-TSH (1 µg/dL) or HTK-ACTH (1 µg/dL) in groups 3 and 4. The solutions were cooled to 4°C-6°C. Within 30 minutes of the first perfusion, the discharged fluid was clear and the kidneys cooled to 4°C. The levels of lactate dehydrogenase, asparagine and alanine aminotransferases, lactates, total protein, potassium, calcium, and pH were determined in the perfusate. After 24 and 48 hours the rinsing procedure and the above-mentioned tests were repeated. Differences between the means of 2 independent samples were tested with a nonparametric Mann-Whitney U test. RESULTS: As the result of hormone addition, in both time intervals it was possible to observe considerably lower protein concentrations (g/L) in perfusates compared with HTK solution, without an addition. At 24 hours, we measured following values: 36 ± 4, 8 ± 3 and 6 ± 1 versus 48 hours, 34 ± 1, 2 ± 1, and 4 ± 1 in groups 2, 3, and 4. A similar pattern was observed with LDH (U/L) at 48 hours: 662 ± 89, 374 ± 151, and 386 ± 111, respectively. Lactate concentrations (mmol/L) were then significantly higher: 1.4 ± 0.3 in the TSH group and 1.2 ± 0.5 in the ACTH group as opposed to 0.2 ± 0.1 in unmodified HTK group. CONCLUSION: We observed the possibility of cytoprotective actions of TSH and ACTH addition to the perfusion fluid during cold ischemia, positive effects that were especially visible upon prolonged 48-hour storage.

Impact of early lymph node procurement to facilitate histocompatibility testing on long-term cadaveric kidney graft survival.

BACKGROUND: Prolonged cold ischemia time (CIT) is a clinically important causes of delayed graft function (DGF) after kidney transplantation. As DGF has been previously shown to have a deleterious influence on long-term graft survival, in the present study we analyzed the impact of early lymph node (LN) procurement on CIT, HLA mismatches, and long-term kidney graft outcome. MATERIALS AND METHODS: We evaluated 394 consecutive cadaveric procedures performed from 2001 to 2006, including 289 recipients, in whom LN were obtained before kidney procurement seeking to shorten the total time for HLA typing and crossmatch procedures. RESULTS: During 58±6 months, 24 patients died (918 [8.3%] in the early and 6 [5.7%] in late procurement group, P=ns) and 52 lost their kidney grafts (31 [10.7%] vs 21 [20%]; P=.025). Early procurement of LN performed in 73.4% of all kidney graft recipients shortened CIT by almost 7 hours (22.9 vs 16.1 hours; P<.001), with a nonsignificantly lower incidence of DGF (32.2% vs 41.0%; P=.13). However, a Cox proportional hazards regression model revealed that early procurement reduced the risk of death-censored kidney graft loss by roughly 40% (log-rank, P=.013). CONCLUSION: Early LN procurement in significantly shorten CIT and subsequently reduced the risk of long-term kidney graft loss.