Abstract: Many studies have indicated that nuclear DNA content evaluation can be used to predict biological behavior of transitional cell carcinoma (TCC) of the bladder. Some authors also indicated that static cytometry is more useful in DNA content analysis than flow cytometry. The aim of the present study was to evaluate the prognostic significance of DNA ploidy in TCC of the bladder, performed by using static cytometry with an image analyzer, and monitoring patients at 10 years follow-up. Thirty-one consecutive patients underwent transurethral or open surgery for bladder tumors, and neoplastic tissue samples taken from each patient were imprinted on glass slides and sent for histopathological and DNA content evaluation. DNA ploidy evaluation was performed using a CAS 200 image analyzer. Nuclear DNA content evaluation was compared to patient follow-up on recurrence, progression or survival performed 10 years after surgery. Pathological evaluation demonstrated the presence of superficial TCC in 23 patients, while 8 had an invasive bladder tumor. Twenty-nine tumor samples were adequate for DNA content measurement, with 13 showing diploid DNA content and 16 with aneuploid DNA content. At 10 years follow-up, all patients with aneuploid DNA content demonstrated a lower survival time (p=0.049) and higher recurrence rate (p=0.0346). A log-rank test demonstrated that stage, grade and nuclear DNA content are the most useful prognostic parameters for predicting the biological behavior of TCC of the bladder. These results confirm that static cytometry is a good and reliable method to evaluate DNA tumor content and considered a useful prognostic parameter for predicting recurrence rate, disease progression or survival in patients affected by bladder tumors.

Abstract: OBJECTIVE: The aim of this pilot study is to analyze the safety and short-term efficacy of gemcitabine (GEM) as salvage intravesical therapy in a very selected population of bacille Calmette-Guérin (BCG)-resistant T1G3 patients. METHODS: 9 recurrent BCG-refractory pT1G3 patients, unsuitable for radical treatment, were treated with GEM, and compared with 10 pT1G3 patients previously treated with at least two courses of transurethral resection plus BGC, with further conservative endovesical BCG administration. RESULTS: Both intravesical administrations of GEM and BCG were generally well tolerated: no severe adverse events were reported. Of the 9 patients treated with GEM, 3 were recurrence-free after 13, 17 and 21 months and 7 kept an intact bladder, with an overall survival rate of 9 of 9. Among 10 patients treated with BCG instillation, 1 was recurrence-free after 27 months and 6 kept their bladders, with a survival rate of 8 of 10. CONCLUSIONS: Our experience confirms the high risk of tumor recurrence and progression of BCG-refractory pT1G3 transitional cell carcinoma. In this case, further BCG courses seem to be unsuitable, resulting in a high risk of tumor progression and mortality. The use of GEM in BCG-refractory pT1G3 patients has to be considered experimental until multicentric randomized studies with adequate follow-up are able to confirm the preliminary results of this pilot study.

Abstract: PURPOSE: Several urinary markers have been recently introduced in clinical practice for improving the noninvasive diagnosis of transitional cell carcinoma. Although microsatellite analysis must be considered the best method in terms of results, its cost and method time are unacceptable for daily use. We validated a more rapid and inexpensive method of determination using rapid DNA extraction and automatic multiplex polymerase chain reaction amplification. MATERIALS AND METHODS: A total of 120 patients who presented consecutively to a urological office, including 73 with transitional cell carcinoma and 43 who served as controls, were selected for study. Microsatellite analysis was performed in the blood/urine pair using 3 multiplex polymerase chain reactions per patient. Urine sediment inflammatory cells were assessed by urine dipstick test. Ten microsatellite loci were investigated. Numerical data collected during electrophoresis of the amplified segment in an ABI Prism 310 Genetic Analyzer were used to calculate the cutoff for allelic imbalance. Method sensitivity, specificity, and positive and negative predictive values were calculated. RESULTS: A total of 66 patients had microsatellite analysis alterations in urine sediment, of whom 59 had transitional cell carcinoma, while 7 had other urological diseases. Test sensitivity and specificity were 80.8% and 85.1%, respectively. Statistical analysis did not indicate any significant influence of inflammatory status on microsatellite analysis diagnostic performance. In the control group the allelic imbalance on chromosome 9 was significantly lower than on other chromosomes (p = 0.0143). This could confirm that chromosome 9 has a specific role in transitional cell carcinoma. The multiplex microsatellite analysis method was low cost and not time-consuming. CONCLUSIONS: Multiplex microsatellite analysis is a noninvasive, rapid, inexpensive and reproducible method for screening for and monitoring superficial transitional cell carcinoma. It should be considered an alternative method to urinary cytology and it should also be considered in the presence of urine sediment inflammatory cells.

Abstract: OBJECTIVES: To determine the tolerability and efficacy after 1 year of weekly intravesical gemcitabine therapy in patients with intermediate-risk and high-risk superficial transitional cell carcinoma. METHODS: A total of 116 patients with intermediate-risk and high-risk bladder cancer who had undergone transurethral resection were treated with one cycle (once a week for 6 weeks) of gemcitabine 2000 mg. Local and systemic tolerability and efficacy were evaluated. RESULTS: In terms of the tolerability of gemcitabine, 14 patients (12.0%) reported urgency, 6 (5.1%) dizziness and slight fever (less than 38 degrees C), 1 (0.8%) severe abdominal pain, with ulcerative lesions of the bladder mucosa at cystoscopy, and 1 (0.8%) parosmia. The remaining 94 patients (81.3%) did not report any local side effects during the treatment period. In terms of efficacy, recurrence developed in 29 patients (25.4%) a mean of 7 months after transurethral resection; 85 patients (74.6%) were disease free after 12 months. The univariate analysis showed a greater level of efficacy in patients with a first occurrence (P = 0.0408), patients who had had no previous treatment (P = 0.0368), and patients with Stage pTa superficial transitional cell carcinoma (P = 0.0018). The multivariate analysis did not reveal any significant data. No significant differences were found between the intermediate-risk and high-risk patients in tolerability or efficacy. No recurrence developed in 18 (75%) of 24 intermediate-risk bacille Calmette-Guérin-refractory or 7 (43.7%) of 16 high-risk bacille Calmette-Guérin-refractory patients. CONCLUSIONS: The results of our study have confirmed the good tolerability and 1 year efficacy of intravesical gemcitabine. The treatment schedule proposed resulted in high patient compliance, and the results can be compared with the results of studies using other intravesical treatments.

Abstract: A giant stone in enterocystoplasty is a very rare condition; only 5 cases have been reported in the literature. A 50-year-old female with an enterocystoplasty due to congenital myelomeningocele presented at our institution for an asymptomatic 10-cm bladder stone, incidentally detected during an imaging procedure for uterine fibroma. The patient was treated with a laparoscopic hysterectomy with bilateral annexectomy, neocystotomy and stone removal under general anesthesia. The case report and an accurate literature review are reported. The incidence, risk factors, pathophysiology and treatment options are analyzed.

Abstract: It has recently been shown that allelic abnormalities, detected by microsatellite analysis of the DNA extracted from urine sediment, can be successfully used for the detection of transitional cell carcinoma (TCC) of the bladder. The diagnostic accuracy of urinary cytology, urinary bladder cancer (UBC) marker, bladder tumor antigen (BTA) and microsatellite sequence alterations was compared in 42 patients who were recruited for the study. Of them, 30 had been diagnosed with TCC at cystoscopy plus biopsy (group A). Seven patients without any apparent lesions after trans-urethral resection (TUR) and 6 subsequent weeks of endovesical administration of bacillus Calmette-Guerin (BCG), had irritative symptoms. None of them had positive cytology or TCC bladder mucosa biopsies (group B). In the control group were 5 other subjects who were affected by benign prostatic hypertrophy and candidates for prostatectomy (group C). Urine and blood samples were obtained from all of the patients before surgery. Tumor tissue and normal mucosa samples were taken from groups A and C during surgery. Different urinary sediment analyses were performed by using both nuclear medicine and molecular tests. UBC and BTA-t analyses were carried out using monoclonal antibody tests while microsatellite analyses were performed using extracted DNA and electrophoresis of polymerase chain reaction (PCR) products on 13 different primers. Urinary cytological examinations were carried out using the Autocyte Preparation System(R). Urinary cytology confirmed the presence of TCC in 13.3% of patients. The BTA-t marker allowed the identification of 73.3% of cancers with 50% specificity; the UBC marker identified 63.3% of the cases with 41.6% specificity. Microsatellite analysis permitted the identification of 83.3% of the tumors with 100% specificity. DNA analysis demonstrated high sensitivity in patients affected by superficial (81.4%) or G1 (80%) tumors, even when cytological studies demonstrated little or no sensitivity. Microsatellite analysis is a highly-sensitive and specific marker for TCC diagnosis and its monitoring, especially in patients with low-stage and low-grade tumors. Other testing procedures failed to increase urinary cytological diagnostic significance.

Abstract: OBJECTIVES: To validate an Italian version of the University of California-Los Angeles Prostate Cancer Index (UCLA-PCI). METHODS: Men with prostate cancer treated with radical retropubic prostatectomy (RRP) or external beam radiation (EBR) were retrospectively selected. Each subject was asked to complete the UCLA-PCI short form and the 36-item short-form health survey (SF-36) questionnaires (principal sample); a subgroup completed both questionnaires twice (retest sample). A linguistic translation and validation was performed. Psychometric properties were analyzed. RESULTS: A total of 595 men were included in the study: 394 from the Department of Urology (treated with RRP) and 201 from the Department of Radiotherapy (treated with EBR). Of these, 75 patients (50 from the RRP group and 25 from the EBR group) were selected as the retest cohort. Internal consistency reliability of the SF-36 and UCLA-PCI for the primary sample of the surgery and radiotherapy populations ranged from 0.82 to 0.94; the mean values of sexual function were superior in the principal urology group, whereas the principal radiotherapy group demonstrated superior mean values of urinary function and urinary bother. A strong correlation between urinary function and sexual and bowel function, and between urinary function and all bother scales, was reported only for patients treated with RRP. The expected correlation between sexual function and sexual bother was reported for the EBR group but not for the RRP group. CONCLUSIONS: Our study demonstrated the excellent psychometric properties of the Italian version of the UCLA-PCI; the Italian version of the UCLA-PCI questionnaire will allow cross-cultural comparative studies in men with prostate cancer.

Abstract: Many authors have shown that tumor hypoxia exerts its own influence on malignant progression by inducing angiogenetic factors and new blood vessels inside and around the tumor. This event usually suggests a poor prognosis and/or aggressive tumor behavior. The objective of the present study is to compare molecular analysis of angiogenetic factors with microvessel density (MVD) in bladder carcinoma. Twenty-nine consecutive patients underwent transurethral or open surgery for bladder tumors. Neoplastic tissue samples, normal-appearing bladder mucosa and blood samples were taken from each patient. All the tissues underwent mRNA extraction and Northern blot analysis, marked with specific probes for inducible nitric oxide sinthase (iNOS), cyclo-oxygenase-2 (cox-2), vascular endothelial growth factor (VEGF) and were evaluated by gel-electrophoresis. Microvessel density, a quantitative analysis for neoangiogenesis, was also evaluated by using CD31 immunohistochemical assay and compared with both molecular analysis and patient follow-up. Two follow-up for recurrence and/or progression were performed at 74 months and 10 years from surgery respectively. Pathological evaluation demonstrated the presence of superficial transitional cell carcinoma (sTCC) in 15 patients, while 14 had an invasive bladder tumor (iBT). At both 74 months and 10 years follow-up, all patients with lower MVD had a shorter survival time. No significant results were obtained by comparing disease progression or survival rate with VEGF, iNOS and COX-2 levels. A proportional increase of VEGF expression and MVD compared with poor prognosis was the expected outcome of our study. These results were disregarded at both the 1st and the 2nd follow-up. A strong association between MVD>20 and survival rate was noted both in sTCC (p=0.024) and iBT (p>0.001) patients. These results confirm that MVD could be considered a good prognostic factor. The angiogenetic cytokines overexpression found in control tissue samples of sTCC could have clinical significance, either as a macroscopically unidentified diffuse carcinogenetic process or the presence of a systemic immune-response against tumor cells.