Immunosuppression for liver transplantation in HCV-infected patients: Mechanism-based principles

We retrospectively analyzed 42 hepatitis C virus (HCV)-infected patients who underwent cadaveric liver transplantation under two strategies of immunosuppression: (1) daily tacrolimus (TAC) throughout and an initial cycle of high-dose prednisone (PRED) with subsequent gradual steroid weaning, or (2) intraoperative antithymocyte globulin (ATG) and daily TAC that was later space weaned. After 36 ± 4 months, patient and graft survival in the first group was 18/19 (94.7%) with no examples of clinically serious HCV recurrence. In the second group, the three-year patient survival was 12/23 (52%), and graft survival was 9/23 (39%); accelerated recurrent hepatitis was the principal cause of the poor results. The data were interpreted in the context of a recently proposed immunologic paradigm that is equally applicable to transplantation and viral immunity. In the framework of this paradigm, the disparate hepatitis outcomes reflected different equilibria reached under the two immunosuppression regimens between the relative kinetics of viral distribution (systemically and in the liver) and the slowly recovering HCV-specific T-cell response. As a corollary, the aims of treatment of the HCV-infected liver recipients should be to predict, monitor, and equilibrate beneficial balances between virus distribution and the absence of an immunopathologic antiviral T-cell response. In this view, favorable equilibria were accomplished in the nonweaned group of patients but not in the weaned group. In conclusion, since the anti-HCV response is unleashed when immunosuppression is weaned, treatment protocols that minimize disease recurrence in HCV-infected allograft recipients must balance the desire to reduce immunosuppression or induce allotolerance with the need to prevent antiviral immunopathology. Copyright © 2005 by the American Association for the Study of Liver Diseases

Alemtuzumab pre-conditioning with tacrolimus monotherapy in pediatric renal transplantation

We employed antibody pre-conditioning with alemtuzumab and posttransplant immunosuppression with low-dose tacrolimus monotherapy in 26 consecutive pediatric kidney transplant recipients between January 2004 and December 2005. Mean recipient age was 10.7 ± 5.8 years, 7.7% were undergoing retransplantation, and 3.8% were sensitized, with a PRA >20%. Mean donor age was 32.8 ± 9.2 years. Living donors were utilized in 65% of the transplants. Mean cold ischemia time was 27.6 ± 6.4 h. The mean number of HLA mismatches was 3.3 ± 1.3. Mean follow-up was 25 ± 8 months. One and 2 year patient survival was 100% and 96%. One and 2 year graft survival was 96% and 88%. Mean serum creatinine was 1.1 ± 0.6 mg/dL, and calculated creatinine clearance was 82.3 ± 29.4 mL/min/1.73 m 2. The incidence of pre-weaning acute rejection was 11.5%; the incidence of delayed graft function was 7.7%. Eighteen (69%) of the children were tapered to spaced tacrolimus monotherapy, 10.5 ± 2.2 months after transplantation. The incidence of CMV, PTLD and BK virus was 0%; the incidence of posttransplant diabetes was 7.7%. Although more follow-up is clearly needed, antibody pre-conditioning with alemtuzumab and tacrolimus monotherapy may be a safe and effective regimen in pediatric renal transplantation. © 2007 The Authors

Analysis of Antimicrobial-Triggered Membrane Depolarization Using Voltage Sensitive Dyes

The bacterial cytoplasmic membrane is a major inhibitory target for antimicrobial compounds. Commonly, although not exclusively, these compounds unfold their antimicrobial activity by disrupting the essential barrier function of the cell membrane. As a consequence, membrane permeability assays are central for mode of action studies analysing membrane-targeting antimicrobial compounds. The most frequently used in vivo methods detect changes in membrane permeability by following internalization of normally membrane impermeable and relatively large fluorescent dyes. Unfortunately, these assays are not sensitive to changes in membrane ion permeability which are sufficient to inhibit and kill bacteria by membrane depolarization. In this manuscript, we provide experimental advice how membrane potential, and its changes triggered by membrane-targeting antimicrobials can be accurately assessed in vivo. Optimized protocols are provided for both qualitative and quantitative kinetic measurements of membrane potential. At last, single cell analyses using voltage-sensitive dyes in combination with fluorescence microscopy are introduced and discussed

Chronic liver allograft rejection in a population treated primarily with tacrolimus as baseline immunosuppression: Long-term follow-up and evaluation of features for histopathological staging

Background: Predisposing factors, long-term occurrence, and histopathological changes associated with recovery or progression to allograft failure from chronic rejection (CR) were studied in adult patients treated primarily with tacrolimus. Methods: CR cases were identified using stringent criteria applied to a retrospective review of computerized clinicopathological data and slides. Results: After 1973 days median follow- up, 35 (3.3%) of 1049 primary liver allograft recipients first developed CR between 16 and 2532 (median 242) days. The most significant risk factors for CR were the number (P40 years (P<0.03). Other demographic and matching parameters were not associated with CR in this cohort. Ten patients died with, but not of, CR. Eight required retransplantation because of CR at a median of 268 days. Ten resolved either histologically or by normalization of liver injury tests over a median of 548 days. CR persisted for 340 to 2116 days in the remaining seven patients. More extensive bile duct loss (P<0.01), small arterial loss (p<0.03), foam cell clusters (P<0.01) and higher total bilirubin (p<0.02) and aspartate aminotransferase (p<0.03) were associated with allograft failure from CR. Conclusions: Early chronic liver allograft rejection is potentially reversible and a combination of histological, clinical and laboratory data can be use to stage CR. Unique immunological and generative properties of liver allografts, which lead to low incidence and reversibility of early CR, can provide insights into transplantation biology

Renal homotransplantation in pediatric patients.

Fifty-seven patients were treated with renal homotransplantation from 1½ to 7⅔ years ago; 23 patients were 12 years or younger and the other 34 patients were 13 to 18. Family members (usually parents) were the primary donors in 45 cases. Unrelated volunteers or cadavers donated the other 12 homografts. Immunosuppression was with azathioprine and prednisone, and in some cases also with ALG. Forty-two of the 57 recipients survived for at least 1 year. Additional deaths occurred at 17½ and 19 months leaving 40 recipients (70.2%) alive. Six survivors had successful retransplantation following late failure of their original homografts. Control of rejection was not particularly different than in adult cases. “Homograft glomerulonephritis” was found in chronically tolerated transplants, but no more frequently than in older patients. Many postoperative problems in the pediatric age group were the consequence of retardation of growth caused either by pre-existing uremia or by the need for high dose postoperative steroid therapy, orthopedic accidents such as femoral and vertebral fractures, and psychiatric complications which led to two suicides. In spite of these difficulties, the meaningful rehabilitation that was obtained in the chronic survivors makes us regard pediatric patients as favorable candidates for therapy with renal transplantation

Islet isolation assessment in man and large animals

Recent progress in islet isolation from the pancreas of large mammals including man, accentuated the need for the development of precise and reproducible techniques to assess islet yield. In this report both quantitative and qualitative criteria for islet isolation assessment were discussed, the main topics being the determination of number, volume, purity, morphologic integrity and in vitro and in vivo function tests of the final islet preparations. It has been recommended that dithizone should be used as a specific stain for immediate detection of islet tissue making it possible to estimate both the total number of islets (dividing them into classes of 50 μ diameter range increments) and the purity of the final preparation. Appropriate morphological assessment should include confirmation of islet identification, assessment of the morphological integrity and of the purity of the islet preparation. The use of fluorometric inclusion and exclusion dyes together have been suggested as a viability assay to simultaneously quantitate the proportion of cells that are intact or damaged. Perifusion of islets with glucose provides a dynamic profile of glucose-mediated insulin release and of the ability of the cells to down regulate insulin secretion after the glycemic challenge is interrupted. Although perifusion data provides a useful guide to islet viability the quantity and kinetics of insulin release do not necessarily predict islet performance after implantation. Therefore, the ultimate test of islet viability is their function after transplantation into a diabetic recipient. For this reason, in vivo models of transplantation of an aliquot of the final islet preparation into diabetic nude (athymic) rodents have been suggested. We hope that these general guidelines will be of assistance to standardize the assessment of islet isolations, making it possible to better interpret and compare procedures from different centers. © 1990 Casa Editrice il Ponte

Quantitative Microscopy of Hepatic Changes Induced by Phenethyl Isothiocyanate in Fischer-344 Rats Fed Either a Cereal-Based Diet or a Purified Diet

Hepatic changes induced by phenethyl isothiocyanate (PEITC) in the liver of rats were determined by quantitative microscopy. Groups of male Fischer-344 rats were fed either a standard, cereal-based diet (Wayne rodent meal) or a purified diet (AIN-76A) containing PEITC at concentrations of 0.75 and 6.0 mmol/kg for 13 wk. Severe hepatic lipidosis was observed in control rats fed the purified diet. Addition of PEITC to the purified diet significantly reduced lipid content in hepatocytes. In contrast, lipid content in the liver of the rats fed the cereal-based diet containing PEITC was greater than in control rats maintained on the same diet. In addition, dose-related reductions in hepatocyte, lipid droplet, peroxisome, and mitochondrial volumes were observed in PEITC-treated rats fed the cereal-based diet. These results indicate that PEITC exerts differential effects on the liver of rats fed either the cereal-based or purified diet.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/68493/2/10.1177_019262339502300602.pd

Discrete Source Survey of 6 GHz OH emission from PNe & pPNe and first 6 GHz images of K 3-35

The aim of this study is to investigate the physical properties of molecular envelopes of planetary nebulae in their earliest stages of evolution. Using the 100m telescope at Effelsberg, we have undertaken a high sensitivity discrete source survey for the first excited state of OH maser emission (J=5/2, 2PI3/2 at 6GHz) in the direction of planetary and proto-planetary nebulae exhibiting 18cm OH emission (main and/or satellite lines), and we further validate our detections using the Nan\c{c}ay radio telescope at 1.6-1.7GHz and MERLIN interferometer at 1.6-1.7 and 6GHz. Two sources have been detected at 6035MHz (5cm), both of them are young (or very young) planetary nebulae. The first one is a confirmation of the detection of a weak 6035MHz line in Vy 2-2. The second one is a new detection, in K 3-35, which was already known to be an exceptional late type star because it exhibits 1720MHz OH emission. The detection of 6035MHz OH maser emission is confirmed by subsequent observations made with the MERLIN interferometer. These lines are very rarely found in evolved stars. The 1612MHz masers surround but are offset from the 1720 and 6035MHz masers which in turn lie close to a compact 22GHz continuum source embedded in the optical nebula.Comment: 9 pages, 7 figures, published in A&