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OBJECTIVES: Type 1 diabetes mellitus is an emerging epidemic worldwide and results from autoimmune destruction of insulin-producing ß cells. Islet transplanting is a potential treatment for type 1 diabetes mellitus. MATERIALS AND METHODS: The Shiraz Organ Transplant Center is a leading center for organ transplants, especially pancreatic transplants, in Iran. For this reason, we want to establish an islet transplanting program. Here, we briefly describe our experience with islet isolation on 6 pancreata from deceased donors. We discussed the necessary equipment required for this procedure, as well as the professionals needed and a specially planned facility. RESULTS: Islet yield was ≤ 100 000 (islet equivalent), viability 40% to 45%, and the purity was 30% to 45%. We do not have a refrigerated COBE processor for purification; therefore, the yield was low. Our experience shows that we should improve things, so as to acquire more islets for developing clinical grade cell therapy. CONCLUSIONS: Overall, isolation costs are high, and accessing a safer, more economic, and persistent source of material and reagents will improve this technique.

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BACKGROUND: Hepatocytes are used as an in vitro model to evaluate drug metabolism. Human hepatocyte transplant has been considered as the temporary treatment of acute liver failure. Optimization freezing methods is very important to preserve both cell viability and function which are achieved by cryopreservation mostly always. OBJECTIVES: The present study aimed to investigate the cryoprotective effect of DTT and fructose on primary rat hepatocytes and HepG2 cells. MATERIALS AND METHODS: Both fresh rat hepatocytes and HepG2 cell line were incubated with fructose (100 and 200 mM) and dithiothreitol (DTT) (25, 50, 100, 250, and 500 µM) at 37°C for 1 and 3 hours, respectively. The preincubated hepatocytes were cryopreserved for two weeks. Hepatocytes viability and function were determined post thawing and the results were compared with the control group. RESULTS: The viability of both rat hepatocytes and HepG2 cells were significantly increased after one hour preincubation with fructose 200 mM. Preincubation with DTT (50 µM, 100 µM. 250 µM and 500 µM) improved the viability and function upon thawing in both cell types (P < 0.001). In rat hepatocytes, no significant change was observed in albumin, urea production, and LDH leakage after preincubation with fructose or DTT. In HepG2 cells, albumin and urea production were significantly increased after preincubation with DTT (500 µM, 1 hour). The GSH content was significantly increased in DTT (250 and 500 µM, 1 hour) groups in both rat hepatocyte and HepG2 cells. CONCLUSIONS: Incubation of hepatocytes with fructose and DTT prior to the cryopreservation can increase the cell viability and function after thawing.

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OBJECTIVES: Nitric oxide is a major mediator in vascular biology and regulator of regional blood flow. Its production is catalyzed by the enzyme endothelial nitric oxide synthase. Protective actions of nitric oxide in ischemia and reperfusion are due to its potential as an antioxidant and anti-inflammatory agent, along with its inhibitory effects on cell signaling pathways of nuclear proteins, such as NF-kB. The endothelial nitric oxide synthase gene polymorphisms affect endothelial nitric oxide synthase activity and are associated with endothelial dysfunction. This study sought to examine the association between single nucleotide polymorphisms in endothelial nitric oxide synthase gene (rs 2070744, 27VNTR, and rs1799983) and the development of acute rejection in renal transplant patients. MATERIALS AND METHODS: Sixty-six renal transplant recipients (33 patients with an episode of acute rejection and 33 recipients an episode of acute rejection), between June 2010 and March 2011, were included. The polymorphism was determined by simple polymerase chain reaction and polymerase chain reaction-restriction fragment-length polymorphism analysis. RESULTS: There was only a significant association of endothelial nitric oxide synthase -786T allele and acute rejection (P = .03). Recessive model of T-786C alleles (TT vs TC+CC) and acute rejection confirmed a significant association (odds ratio: 3.12; 95% CI: 0.01-9.83; P = .025). Haplotype CbG was higher in recipients without rejection as compared to rejection group (OR: 0.42, 95% CI: 0.16-1.13; P < .05). Respecting the endothelial nitric oxide synthase gene 894G/T single nucleotide polymorphisms and 27VNTR, no significant association between the allele/genotype and acute rejection was seen. CONCLUSION: Recipient endothelial nitric oxide synthase gene polymorphisms do not alter the risk of acute rejection after a renal transplant. Rejection is a complex immunologic event. Therefore, finding associated genetic variants demands a multicentric larger sample size.

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Management of renal transplant patients requires periodic measurement of renal function especially in early post transplant period. This is usually assessed by measuring the creatinine clearance, but because of its limitations, it is not an ideal marker for assessing the renal function. Serum cystatin C (sCyC) appears to be an endogenous marker of glomerular filtration rate (GFR). To assess the use of sCyC as a marker of renal function in kidney transplant patients, we compared it with serum creatinine (sCr) and 24-hour urine creatinine clearance (CrCl) in the first week post-transplantation. Among 60 patients (62.8% men, 37.2% women) undergoing kidney transplantation (average age: 44.87 +/- 13.37 years), we determined renal function at 1, 3, 5, and 7 days after kidney transplantation using: sCr, sCyC and CrCl in a 24-hours urine specimen. During the first 5 days following transplantation, there was a progressive decline in sCr levels. In the first 5 days, post transplantation we could not find good correlation between CrC and sCyC, and the sCyC increased during these 5 days, but after that in day 7, there was a good correlation between CrC and sCyC which is coinciding with decreasing the dose of steroid (r= .625). Therefore, we recommend using sCyC may be used as a marker of renal function after one-week post kidney transplantation.

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OBJECTIVE: Transplantation of renal grafts is an established treatment for renal failure in a variety of medical conditions. Acute allograft rejection remains an important cause of morbidity after kidney transplantation, and has been shown to be a crucial determinant of long-term graft function. Although rejection is mediated by recipient lymphocytes, both donor and recipient factors contribute to the local environment that influences the severity of rejection response. Because cytokines are the main components of immune responses, we evaluate single nucleotide polymorphisms (SNP) of several cytokine genes that may influence the production of a given cytokine and therefore the features of immune reactions. MATERIAL AND METHODS: The aim of this study was to determine the impact of the cytokine gene polymorphism of pro and anti-inflammatory cytokines on the development of acute allograft rejection, which could be used in pretransplant patient assessment. Three SNPs including IL-10 (-1082 G/A), TNFA (-308 G/A), and INFG (+874 T/A) were analyzed in 46 patients with acute allograft rejection, 54 patients with stable graft function and their kidney donors by PCR-ARMS method. RESULTS: We are unable to find statistically significant association between any of the studies polymorphisms and clinical outcomes. CONCLUSION: We have found no evidence to suggest that either recipient or donor cytokines polymorphisms determine the incidence of acute rejection after renal transplantation. Our observation, however, is based on few cases, and this may mask a possible favorable effect. It is recommended that several functionally related genes should be tested in similar studies, since this approach has a higher chance to detect genetic risk factors than the screening of single genetic variants.

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Hepatitis B virus is a hepatotropic virus that causes acute and chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma; it is responsible for more than one million deaths annually worldwide despite hepatitis B vaccination. Until now, there are eight known genotypes (A-H).Clinical course of chronic hepatitis B varies according to the genotype of Hepatitis B virus. Liver biopsy becomes necessary to judge the degree of liver lesions and to make the final diagnosis, especially to make the diagnosis for latent liver damage and early-compensated cirrhosis. Genotype is very important for prognostication, but it has not yet been reported on liver tissues. Sometimes, it can be helpful to do genotyping of Hepatitis B virus of the liver tissue; such conditions include research programs, when serum is not available or when serum is negative for Hepatitis B virus DNA. In this study, we wanted to evaluate the feasibility of a simple method for genotyping of liver tissue samples. We performed genotyping of the liver biopsies and intended to find out a simple and reliable method for genotyping in the paraffin- embedded formalin- fixed liver tissue.Genotype D was the only isolated genotype in all the liver biopsies. The tissue genotype was just the same as that found in serum. The procedure was easy and good for large scale studies.Genotyping in the paraffin-embedded formalin-fixed stored liver tissue can be done with the same accuracy of the serum.

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