Chronic kidney disease after liver, cardiac, lung, heart–lung, and hematopoietic stem cell transplant

Patient survival after cardiac, liver, and hematopoietic stem cell transplant (HSCT) is improving; however, this survival is limited by substantial pretransplant and treatment-related toxicities. A major cause of morbidity and mortality after transplant is chronic kidney disease (CKD). Although the majority of CKD after transplant is attributed to the use of calcineurin inhibitors, various other conditions such as thrombotic microangiopathy, nephrotic syndrome, and focal segmental glomerulosclerosis have been described. Though the immunosuppression used for each of the transplant types, cardiac, liver and HSCT is similar, the risk factors for developing CKD and the CKD severity described in patients after transplant vary. As the indications for transplant and the long-term survival improves for these children, so will the burden of CKD. Nephrologists should be involved early in the pretransplant workup of these patients. Transplant physicians and nephrologists will need to work together to identify those patients at risk of developing CKD early to prevent its development and progression to end-stage renal disease

Diversity of peripheral blood mononuclear cells as revealed by a novel multiple microgel 'comet assay'

Multiple microgel comet assay (MMCA) is a methodological adaptation of the single-cell gel electrophoresis assay in which we have introduced the use of standard agarose plug molds in an attempt to improve and expand the applications of the assay. We focused on the study of the heterogeneity of peripheral blood mononuclear cells (PBMC) at the level of the basal single-strand breakage and the DNA damage induction caused by ionizing radiation. Differences among subpopulations were also investigated at the level of chromatin organization and methylation after Notl digestion of microgel-embedded cells. In parallel experiments, the Notl-digested nucleoids were also analyzed with the use of pulsed-field gel electrophoresis (PFGE) and the DNA migration patterns were compared with the corresponding patterns from the MMCA. Significant heterogeneity in the distribution of the oxidative DNA damage, as well as intracellular variations in the Notl digestion patterns were observed in the cell population of PBMC. The combined use of both the comet assay and PFGE provides a useful model for analysis of variation in DNA damage in individual cells as well as information on size of DNA fragments. (C) 2000 Wiley-Liss, Inc

Diversity of peripheral blood mononuclear cells as revealed by a novel multiple microgel 'comet assay'

Multiple microgel comet assay (MMCA) is a methodological adaptation of the single-cell gel electrophoresis assay in which we have introduced the use of standard agarose plug molds in an attempt to improve and expand the applications of the assay. We focused on the study of the heterogeneity of peripheral blood mononuclear cells (PBMC) at the level of the basal single-strand breakage and the DNA damage induction caused by ionizing radiation. Differences among subpopulations were also investigated at the level of chromatin organization and methylation after Notl digestion of microgel-embedded cells. In parallel experiments, the Notl-digested nucleoids were also analyzed with the use of pulsed-field gel electrophoresis (PFGE) and the DNA migration patterns were compared with the corresponding patterns from the MMCA. Significant heterogeneity in the distribution of the oxidative DNA damage, as well as intracellular variations in the Notl digestion patterns were observed in the cell population of PBMC. The combined use of both the comet assay and PFGE provides a useful model for analysis of variation in DNA damage in individual cells as well as information on size of DNA fragments. (C) 2000 Wiley-Liss, Inc

Effect of cyclosporine therapy with low doses of corticosteroids on idiopathic nephrotic syndrome

Cyclosporine (CyA) has an immunosuppressive effect that might suggest a therapeutic role in idiopathic glomerular conditions. We focused on the optimization of CyA treatment control in patients with idiopathic nephrotic syndrome by using trough-level CyA measurements (C0) and the 2-h postdose levels (C2). Twenty-two patients (14 male, 8 female) with idiopathic nephrotic syndrome and the mean age of 51 ± 18 months (mean [M] ± standard deviation [SD]) were enrolled in our study during a period of 10 months (range: 3-18 months). All of the patients received CyA (2-3 mg/kg) in combination with methylprednisolone. In the present study protocol CyA concentrations (C0, C2), renal function, lipid profile, and degree of proteinuria were determined. The mean proteinuria of our patients before treatment was 11 972 ± 7953 mg/24 H (±SD) and the mean creatinine level (Cr) was 0.99 ± 0.37 mg/dL (±SD). Proteinuria decreased significantly already from the first month of therapy with CyA to 3578 ± 2470 mg/24 H (M± SD), and during the whole study period this reduction was significant (0.56 ± 0.37 gr/24 H (M ± SD), P < 0.05). At the same time renal function preserved, 1.09 ± 0.48 mg/dL (M ± SD). The blood levels of C0 were 135.10 ± 97.36 ng/mL (M ± SD) and the blood levels of C2 were 725 ± 256 ng/mL (M ± SD) at the first month of therapy. At the same time renal function preserved, 1.09 ± 0.48 mg/dL (M ± SD). Total cholesterol levels reduced significantly during study period (276.89 ± 45.57 to 200.67 ± 40.27 mg/dL [M ± SD]). The mean number of antihypertensive medication remained the same. The whole therapeutic protocol did not provoke any kind of side effects and CyA was quite tolerated by our patients. Treatment of idiopathic nephrotic syndrome with low doses of CyA with methylprednisolone leads to remission of proteinuria without deterioration of renal function. Blood levels of C0 for monitoring and treatment of nephrotic syndrome agrees with recent literature, while our study focus on establishing the proper levels of C2 for the treatment of nephrotic syndrome. The efficacy of CyA is combined with safety and tolerance