Conventional in situ hybridization (CISH) can be used for detection of human papillomavirus (HPV) DNA, enabling preservation of the tissue morphology and assessment of the physical state of viral DNA, but has low sensitivity. This study compared the sensitivity and efficiency of in situ hybridization with tyramide signal amplification (ISH TSA) with those of CISH. The HPV status of 77 cases with early stage cervical carcinoma was evaluated with CISH, using biotinylated probes for HPV types 6/11, 16/18 and 31/33/51, and with ISH TSA using probes for HPV types 6/11, 16/18 and 31/33 or 31/33/51. The HPV DNA was detected in 26 (33.8%) cases using CISH, and in 45 (58.4%) cases using ISH TSA. By adding the TSA step, the sensitivity of CISH was enhanced by 24.7%, thus enabling detection of 20 new HPV-positive cases. Multiple HPV infections were detected in four cases. A dot signal pattern was present in 68.9% (31/45) and more than five positive nuclei per sample were found in 82.2% (37/45) of the cases. We found that the ISH TSA system is a fast and simple method for detection of HPV DNA in cervical carcinoma compared to CISH, and is more sensitive and efficient in the detection and typing of HPV, assessment of HPV DNA physical state and evaluation of the number of positive cells than CISH.

We show that our results with urinary diversion are promising in patients requiring radical cystoprostatectomy. The two methods preferred in our institution offer a sufficient protection of the upper urinary tract with a low complication rate, good voiding function and continence.

706 Cobalamin-dependent methionine synthase and related metabolites in mouse colon tumour model mBlackburnl, M. C. Bibby2, A. Nicolaoul

Aggressive Non-Hodgkin lymphoma (NHL) is heterogeneous group with respect to clinical, histopatological and evaluative features. The International Prognostic Index (IPI) has provided a widely accepted prognostic set of criteria to design therapy. However, IPI dose not determine whether routine phenotypic features, beside IPI, may influence survival. We evaluate the prognostic significance of Bcl-2 and Ki67 protein expression in relation to clinical presentation and outcome. In the period 1989–2002,two hundred eleven patients with newly diagnosed aggressive NHL were recorded in our Department. In this study we included patients with available biopsy sample. Protein expression was analyzed on paraffin embedded tumor tissue by imunohistochemistry in relation to clinical factor and outcome. The expression of more than 20% of neoplastic cell was considered positive for bcl-2, and more than 60% for Ki-67. Comparison was made by x2 test. Survival curves were considered by Kaplan-Meyer method and compared by the long-rank test. Sixty seven patients were recorded and their characteristic were: median age 53 years, stage III and IV, 67%; present B symptoms 44%,; bone marrow infiltration 29%, elevated LDH 58%, performas status more then 1– 26%; extra nodal sites more than 2 –34%. All 62 patients received antracyclin based combination chemotherapy, 72% CHOP regiment. Overall survival was 52% with a follow up 6–183 months. Bcl-2 expression was observed in 35,5% (22pts) and Ki67 positivity over 60% in 20 pts (30%). Overall survival was influenced by bcl-2 and Kli67 expression. Bcl-2 positive cases were significantly associated with lower overall survival (20% vs. 58%; p<0.01). Overall survuval was significantly worse in patients with Ki67 positivity over 60% (27% vs. 49%; p<0.05) Bcl-2 and Ki67 monoclonal antibody immunostaging appears to be a simple and reproducible method of determining biological potential of tumor cells and provides useful prognostic information in patients with aggressive NHL.