Sunjay Jain

Apoptosis and caspase-3 in long-term renal ischemia/reperfusion injury in rats and divergent effects of immunosuppressants.

Background. Caspase-3 plays a key role in apoptosis, but the involvement of apoptosis and caspase-3 in mediating long-term ischemia/reperfusion (I/R) and immunosuppressive injury are not fully defined. The present study was undertaken to investigate apoptosis and caspase-3 in a renal I/R injury rat model with or without immunosuppression. Methods. The right renal pedicle was clamped for 45 minutes and left nephrectomy was induced. Cyclosporin A (CsA), tacrolimus (Tac), rapamycin (Rap), or mycophenolate mofetil (MMF) were administered daily. Animals were killed at 16 weeks, and the levels of apoptosis (with in situ end-labeling fragmented DNA), caspase-3 protein (with immunohistochemistry, Western blotting, and activity assay), and messenger RNA (mRNA; with quantitative reverse-transcriptase polymerase chain reaction) were evaluated. Results. Kidneys with I/R injury showed increased apoptosis in tubular and interstitial areas compared with control kidneys. Tacrolimus, Rap, and MMF significantly reduced apoptosis, but CsA did not. Distribution of full-length caspase-3 widened in I/R–injured kidneys from normal distal tubules and collecting ducts to dilated proximal tubules and expanded interstitium, whereas active caspase-3 was mainly scattered in damaged tubules and interstitium. Active caspase-3 staining and 24-kDa active caspase-3 protein was enhanced in I/R–injured and CsA-treated kidneys, but decreased by Tac, Rap, and MMF. These results were also consistent with changes in caspase-3 activity. Although caspase-3 mRNA levels were significantly increased in uninephrectomy and I/R–injured kidneys, they were not significantly affected by the immunosuppressants. In addition, all changes detected were positively correlated with renal structure and function. Conclusion. Apoptosis and caspase-3 are not only involved in the long-term renal I/R injury, but also mediate the divergent effects of immunosuppression in this model.

Rapamycin reduces expression of fibrosis-associated genes in an experimental model of renal ischaemia reperfusion injury

CHRONIC ALLOGRAFT nephropathy (CAN) remains a major cause of renal transplant failure. It is characterised by a progressive functional deterioration, which is associated histologically with graft fibrosis. Cyclosporin has no impact on the development of CAN, and its nephrotoxicity is a contributor to the functional and histological deterioration found in this condition. The new immunosuppressive agent rapamycin inhibits growth factordriven proliferation in a number of cells crucial to the development of fibrosis including smooth muscle cells, endothelial cells, and fibroblasts. It therefore has a potential role in the therapy of CAN. In this experiment, to further explore this role both cyclosporin and rapamycin were administered to a rodent model of renal ischaemia reperfusion injury, which has previously been shown to be associated with up-regulation of fibrosis-associated genes.

Sub‐clinical acute rejection detected using protocol biopsies in patients with delayed graft function

Abstract Acute rejection in renal transplants is difficult to diagnose when patients have delayed graft function (DGF) in the early post‐transplant period. In this study protocol, renal transplant biopsies were performed in an attempt to detect sub‐clinical acute rejection episodes. Eighty‐three patients were eligible for the study, of whom 33 had DGF. All had protocol renal transplant biopsies performed under ultrasound control at 7 days post‐transplant, and those with DGF had further biopsies weekly until the graft functioned. All histologically confirmed acute rejection episodes were treated. Sub‐clinical acute rejection was detected in 6/33 (18%) patients with DGF compared to 2/50 (4 %) in the other patients (P < 0.05). Borderline rejection was present in 4/33 (12 %) and 4/50 (8 %) patients, respectively. Because of the high detection rate of sub‐clinical acute rejection and the low morbidity of renal transplant biopsies, their use is recommended in patients with DGF.

AN AUDIT OF IMPLANTED PENILE PROSTHESES IN THE UK

Associate Editor

Sequential protocol biopsies from renal transplant recipients show an increasing expression of active TGF β

Abstract Chronic allograft nephropathy (CAN) is a major cause of graft loss after renal transplantation. Implicated in the pathogenesis of this complication is overproduction of the cytokine transforming growth factor beta (TGF β). In this study we measured changes in CANs expression in stable patients early after transplantation, and studied links with established risk factors for CAN, such as delayed graft function, acute rejection, and cyclosporine exposure. We took biopsies from 40 renal allografts at time of transplantation (pre‐perfusion), and then, using ultrasound guidance, at 1 week and 6 months after transplantation. An immunofluorescence technique was used to stain sections for active TGF β. These were then assessed by semi‐quantitative scanning laser confocal microscopy. There was very little variation in active TGF‐β expression among patients in their pre‐perfusion biopsies. Expression had increased by 1 week and then very significantly by 6 months (P<0.0001). Patients who suffered delayed graft function had increased TGF‐β expression at both time points. There was no difference regarding donor type, acute rejection, and immunosuppressive drug (cyclosporine or tacrolimus). There was no correlation between the amount of TGF‐β expression at any time‐point and isotope glomerular filtration rate (GFR) at 12 months. This study demonstrated that in a group of stable renal allograft recipients, TGF‐β expression in the kidney increased after transplantation. As the study used protocol biopsies, this increase is unlikely to be due to acute events, and probably represents a genuine increase.

Caspase-7, Fas and FasL in Long-Term Renal Ischaemia/Reperfusion and Immunosuppressive Injuries in Rats

Background/Aims: Ischaemia/reperfusion (I/R) injury is important in kidney transplantation. We have previously demonstrated that long-term I/R injury and immunosuppression affect apoptosis and inflammation, but the underlying mechanisms are far from clear. In this study, the involvement of caspase-7, Fas and FasL was further investigated. Methods: The right renal pedicle was clamped for 45 min followed by left nephrectomy in 40 rats. Cyclosporine (CsA), tacrolimus (Tac), rapamycin (Rap) or mycophenolate mofetil (MMF) were administered daily for 16 weeks. Caspase-7, Fas and FasL expression, and their correlations with caspase-3, apoptosis, inflammation, renal structure and function were evaluated. Results: Active caspase-7 was significantly increased in I/R and CsA-treated kidneys and decreased by Tac, Rap and MMF, while the caspase-7 precursor was enhanced by Rap. Active caspase-7-stained cells were scattered throughout the tubulointerstitium and often had apoptotic features. Fas, but not FasL, was increased in I/R and CsA-treated kidneys and decreased by Rap and MMF. Fas and FasL proteins were mainly located in dilated tubules. There were close correlations among caspase-7, Fas, caspase-3, apoptosis, inflammation, renal structure and function. Conclusion: Caspase-7, associated with caspase-3, apoptosis and inflammation, might be involved in long-term I/R and immunosuppressive injury, at least in part through the Fas-signalling pathway.

The role of photodynamic diagnosis in the contemporary management of superficial bladder cancer

5-ALA is generally administered intravesically 2 h before cystoscopy through a urethral catheter. The procedure requires special telescopes and a specific light source (D-Light, Karl Storz, Germany). Using a foot pedal or a push-button on the camera it is possible to switch between white or blue light during cystoscopy and resection. Papillary tumours appear intensely red when viewed under blue light and red mucosal patches may represent carcinoma in situ (CIS) (Fig. 2).

Surgical Atlas Insertion of an inflatable penile prosthesis

Penile prostheses were first developed 30 years ago [1]; despite ongoing advances in the pharmacotherapy for erectile dysfunction (ED), prosthetic surgery continues to occupy an important therapeutic role and ≈ 1% of all patients with ED eventually have a penile prosthesis inserted. This equates to ≈ 25 000 penile prostheses implanted annually worldwide. In Europe the ratio of semi-rigid to inflatable penile prostheses (IPPs) is 2 : 3, as a result of fiscal restraints by different healthcare systems because patients prefer the cosmetic qualities of IPPs. American Medical Systems (AMS, Minnetonka, NI, USA) and Mentor Medical Ltd. (Santa Barbara, CA, USA) manufacture both types of devices.

Simulation in urology to train non-technical skills in ward rounds

To report our experience of an exercise designed to train newly appointed urology trainees in non‐technical skills on ward rounds as a part of a simulation ‘boot camp’, through a qualitative analysis of participant feedback on the utility of this method of training.