Kesavan Esuvaranathan

Predictive value of p53 and pRb expression in superficial bladder cancer patients treated with BCG and interferon‐alpha

Nuclear p53 and retinoblastoma protein (pRb) were reported to be poor prognostic indicators for transitional cell carcinoma of the bladder. The authors sought to determine the prognostic value of nuclear p53 and pRb in superficial bladder transitional cell carcinoma patients who were treated with intravesical bacille Calmette‐Guerin (BCG) or BCG with interferon‐alpha (IFN‐α).

Stones in Horseshoe Kidneys: Results of Treatment by Extracorporeal Shock Wave Lithotripsy and Endourology

We report the results of 7 patients with calculi in a horseshoe kidney treated by extracorporeal shock wave lithotripsy (ESWL, 9 renal units) and percutaneous nephrolithotripsy (3 renal units) during a 3-year period. In the ESWL only group complete stone clearance was achieved in only 3 patients (50%) after an average of 3 sessions of therapy. On the other hand, complete stone clearance was achieved by percutaneous nephrolithotripsy with minimal complications. The poorer results with ESWL were due to difficulty in ultrasonographic localization of stones as well as poor drainage in these abnormal kidneys. Our experience with the Edap LT01 and the Sonolith 2000 lithotriptors suggests that while reasonable results are possible, treatment probably will require multiple sessions and the eventual outcome is less predictable than in normal kidneys. In contrast, the treatment of complicated stones in a horseshoe kidney presents no additional difficulty.

A Community Based Study of Prostatic Symptoms in Singapore

PURPOSE We describe the prevalence of prostatic symptoms, bother and related quality of life, as well as health seeking behavior in men 40 years old or older in Singapore. MATERIALS AND METHODS A community based cross-sectional study was conducted in men 40 years old or older in Queenstown, Singapore. The International Prostate Symptom Score for benign prostatic hyperplasia was used to score symptom severity objectively. Results obtained were compared to those from United States, Scottish and Japanese populations. RESULTS The prevalence of moderate to severe symptomatology was 10% after age adjustment to match the 1990 Singapore population. The most prevalent symptoms were frequency (22.5% of cases) and nocturia (21.5%) with consistently lower prevalence for bother (6.9% for frequency and nocturia). The prevalence of prostatic symptoms was approximately 3 or more times less than in the Scottish, United States and Japanese populations, while the prevalence of bother was approximately 10 times less. For symptomatic individuals there was poor correlation between symptom severity and bother scores. Bother scores correlated better with quality of life scores (r = 0.50) and were more closely associated with health seeking behavior (p = 0.03) than symptom severity scores (r = 0.39, p = 0.07). CONCLUSIONS The prevalence of prostatic symptoms, severity and bothersomeness were all relatively low in Singapore. Bother was not analogous to symptom severity and should be considered independently in clinical decision making.

Antitumour Immunity of Bacillus Calmette-Guerin and Interferon Alpha in Murine Bladder Cancer

Intravesical Bacillus Calmette-Guerin (BCG) immunotherapy is currently the optimal choice for aggressive superficial bladder cancer, with a 70% response rate. This study investigated whether the antitumour response elicited by BCG could be improved by the addition of recombinant interferon alpha (IFN alpha) in the subcutaneous murine MB49 bladder tumour model. The combination of BCG and IFN alpha had superior and earlier antitumour activity than BCG alone for MB49 cells in culture. A total of 14/15 BCG plus interferon-treated mice and 8/16 BCG-treated mice became tumour free after treatment. BCG or the combination treatment significantly raised the T-helper 1 (Th1) cytokine IFN gamma levels compared with levels in all other groups. Whilst BCG therapy alone increased CD4+ and CD8+ populations in spleens, the combination of BCG and IFN alpha also increased alpha beta+ T cells significantly. Our results suggest that the combination of BCG and IFN alpha may represent a more efficacious therapeutic than BCG alone for superficial bladder cancer.

EFFECTS OF BACILLUS CALMETTE-GUERIN AND INTERFERON alpha-2B ON CYTOKINE PRODUCTION IN HUMAN BLADDER CANCER CELL LINES

PURPOSE To determine the effects of live BCG, autoclaved BCG and interferon alpha-2b on cytokine production in human bladder cancer cell lines. MATERIALS AND METHODS The release of nine cytokines from the human bladder cancer cell lines, RT4, RT112, SD, MGH and J82, was measured by ELISA assay. The mRNA level of IL-6 and GM-CSF was determined by RT-PCR. RESULTS BCG and/or interferon alpha-2b differentially increased IL-1beta, IL-6, IL-8, GM-CSF and TNF-alpha production in the bladder cancer cells. High grade cell lines were more responsive to BCG whereas low grade lines were more sensitive to interferon alpha-2b. This correlated with cytotoxicity and growth inhibition induced by these agents. BCG could also induce low levels of IFN-alpha production in all the cell lines. Compared with live BCG, autoclaved BCG had no antiproliferative effect on MGH cells and was less effective in stimulating the production of IL-6, IL-8 and GM-CSF. However, autoclaved BCG was as effective as live BCG in inhibiting growth and stimulating IL-6 and TNF-alpha production of J82 cells. The combination of BCG and interferon alpha-2b also completely suppressed TGF-beta1 production in the MGH and RT112 cell lines. CONCLUSIONS The combination of BCG and interferon alpha-2b has additive effects in cytokine production from bladder cancer cells. This correlates with cytotoxicity and growth inhibition induced by these agents.

Effects of bacillus Calmette-Guérin and interferon-α-2B on human bladder cancer in vitro

The cytolytic and anti‐proliferative effects of bacillus Calmette‐Guérin (BCG) and/or interferon‐α‐2b (IFN‐α‐2b) on 5 human bladder carcinoma cell lines, RT4, RT112, MGH, SD and J82, were determined. The cell lines showed different sensitivities to BCG and IFN‐α‐2b. BCG had direct dose‐dependent cytolytic and anti‐proliferative effects on MGH, J82 and SD (grade 3 cell lines), whereas RT4 and RT112 (grades 1 and 2, respectively) were less sensitive. Surprisingly, higher concentrations of BCG enhanced cell growth of RT4. IFN‐α‐2b also had cytolytic and anti‐proliferative effects on all 5 cell lines. Thus, the RT4 and RT112 cell lines that were not sensitive to BCG were highly sensitive to IFN‐α‐2b. A combination of BCG and IFN‐α‐2b had additive anti‐proliferative effects on MGH, J82 and RT112. Interleukin‐6 (IL‐6) and tumor necrosis factor‐α (TNF‐α) production by these 5 cell lines was measured after stimulation with BCG and/or IFN‐α‐2b, by ELISA immunoassays. Production of IL‐6 and TNF‐α was significantly increased in MGH and J82 cell lines by the combination of BCG and IFNα‐2b. The enhanced cytolytic and anti‐proliferative effects of the combination of BCG and IFN‐α‐2b may be related to the induction of cytokines.Int. J. Cancer 71: 851‐857, 1997.

Metabonomic Profiling of Bladder Cancer

Early diagnosis and life-long surveillance are clinically important to improve the long-term survival of bladder cancer patients. Currently, a noninvasive biomarker that is as sensitive and specific as cystoscopy in detecting bladder tumors is lacking. Metabonomics is a complementary approach for identifying perturbed metabolic pathways in bladder cancer. Significant progress has been made using modern metabonomic techniques to characterize and distinguish bladder cancer patients from control subjects, identify marker metabolites, and shed insights on the disease biology and potential therapeutic targets. With its rapid development, metabonomics has the potential to impact the clinical management of bladder cancer patients in the future by revolutionizing the diagnosis and life-long surveillance strategies and stratifying patients for diagnostic, surgical, and therapeutic clinical trials. An introduction to metabonomics, typical metabonomic workflow, and critical evaluation of metabonomic investigations in identifying biomarkers for the diagnosis of bladder cancer are presented.

The signalling pathway for BCG-induced interleukin-6 production in human bladder cancer cells

Intravesical bacillus Calmette-Guerin (BCG) is currently the therapy of choice for superficial bladder cancer with a 60-70% response rate. Induction of cytokine production (e.g. IL-6, etc.) by BCG has been found in patients urine in vivo as well as bladder cancer cell lines. However, the signalling mechanisms are still unclear. In this study, we investigated the effect of BCG on cAMP production and its role in regulating interleukin-6 expression in the human bladder cancer cell line, MGH. After 1 hr exposure to BCG, IL-6 gene expression in MGH cells increased by 2.5-3-fold and cAMP production increased by 8-10-fold in a time- and dose-dependent manner. BCG-induced cAMP production was inhibited by both antifibronectin antibody and an adenylate cyclase inhibitor, SQ22536 in a dose-dependent way. In the presence of SQ22536, IL-6 expression in MGH cells was also greatly reduced. Furthermore, cAMP-dependent kinase inhibitors H7 and HA1004 also inhibited BCG-induced IL-6 expression in MGH, with HA1004 being much less effective than H7. Thus, BCG induces cAMP production and may regulate interleukin-6 expression partially via a cAMP-dependent pathway in human bladder cancer cells.

NRAMP1 and hGPX1 Gene Polymorphism and Response to Bacillus Calmette-Guérin Therapy for Bladder Cancer

BACKGROUND The natural resistance-associated macrophage protein 1 (NRAMP1) gene is associated with susceptibility to Mycobacterium tuberculosis in humans and to bacillus Calmette-Guérin (BCG) in mice. The detoxification enzyme, human glutathione peroxidase 1 (hGPX1), is associated with recurrence of bladder cancer (BCa). OBJECTIVE To determine whether NRAMP1 and hGPX1 gene polymorphisms correlate with response to BCG immunotherapy for non-muscle-invasive BCa (NMIBC). DESIGN, SETTING, AND PARTICIPANTS DNA was obtained from the peripheral blood of 99 NMIBC patients who were prospectively randomized to receive postresection intravesical BCG (81 mg [n=50] or 27 mg [n=19]) or BCG (27 mg) with interferon alpha (IFN-α; n=30). The median follow-up time was 60 mo. INTERVENTION Intravesical BCG or BCG-IFN-α. MEASUREMENTS Restriction fragment length polymorphism (RFLP) analysis was performed to identify polymorphisms in the NRAMP1 promoter region (GT repeat number) and at position 543 (aspartate [D] and/or asparagine [N] expression) within the NRAMP1 protein (D543N) and position 198 (proline and/or leucine expression) within the hGPX1 protein (Pro198Leu). Data were analyzed using χ(2) analysis, multivariate analysis, and Kaplan-Meier curves. RESULTS AND LIMITATIONS On univariate analysis, the NRAMP1 D543N G:G genotype had decreased cancer-specific survival (CSS; p=0.036). The hGPX1 CT genotype (Pro-Leu) had decreased recurrence time (p=0.03) after BCG therapy. On multivariate analysis, patients with the NRAMP1 D543N G:G genotype and allele 3 (GT)n polymorphism had decreased recurrence time (p=0.014 and p=0.03) after BCG therapy. The limitation of this study was its small sample size. CONCLUSIONS Polymorphisms of the NRAMP1 and hGPX1 genes may be associated with recurrence of BCa after BCG immunotherapy.

Does Transurethral Laser Ureterolithotripsy Justify its Cost

A prospective consecutive series of 64 patients who underwent transurethral laser ureterolithotripsy using a 7.2F semirigid ureteroscope was compared to the immediately preceding consecutive series of 98 patients who had undergone ultrasound lithotripsy using rigid 9.5F or 12.5F ureteroscopes. The distribution of the calculi by size and composition in both series was similar. There was a higher proportion of upper ureteral calculi in the laser lithotripsy series. The success rate for a first attempt at laser lithotripsy was 92.2% versus 71.4% for the ultrasound series (p less than 0.01). When the stone could be reached ultrasound and laser lithotripsy had a fragmentation rate of 97%. The principal reason for the difference in results was the poorer ability to reach calculi when using the larger rigid ureteroscopes. One patient who had failed ultrasound lithotripsy was successfully treated with laser lithotripsy a year later. The overall morbidity was less for laser lithotripsy. The 3-year cost-benefit analysis revealed a smaller difference in cost than expected and the 5-year analysis was advantageous for laser lithotripsy because of its higher success rate. Savings were also realized in the laser series because of the higher proportion of subjects treated as outpatients, and a lower mean duration of hospitalization and time missed from work. For our center with an annual work load of approximately 100 cases laser lithotripsy achieved a superior cost-benefit ratio.