Identification and validation of the methylated TWIST1 and NID2 genes through real-time methylation-specific polymerase chain reaction assays for the noninvasive detection of primary bladder cancer in urine samples.

BACKGROUND: Accumulating evidence suggests that DNA methylation markers could serve as sensitive and specific cancer biomarkers. OBJECTIVE: To determine whether a panel of methylated genes would have the potential to identify primary bladder cancer (BCa) in voided urine samples. DESIGN, SETTING, AND PARTICIPANTS: A pharmacologic unmasking reexpression analysis in BCa cell lines was initially undertaken to unveil candidate methylated genes, which were then evaluated in methylation-specific polymerase chain reaction (MSP) assays performed on DNA extracted from noncancerous and cancerous bladder tissues. The most frequently methylated genes in cancerous tissues, with 100% specificity, were retained for subsequent MSP analysis in DNA extracted from urine samples to build and validate a panel of potential methylated gene markers. Urine samples were prospectively collected at three urologic centres from patients with histologically proven BCa and processed for use in real-time MSP and cytologic analysis. Patients with nonmalignant urologic disorders were included as controls. MEASUREMENTS: A urine sample was classified as valid when > or = 10 copies of the gene encoding ß-actin were measured in the urine sediment genomic DNA. Sensitivity, specificity, and predictive values of the MSP and cytology tests were assessed and compared. RESULTS AND LIMITATIONS: MSP assays performed on 466 of the 496 (94%) valid urine samples identified two genes, TWIST1 and NID2, that were frequently methylated in urine samples collected from BCa patients, including those with early-stage and low-grade disease. The sensitivity of this two-gene panel (90%) was significantly better than that of cytology (48%), with comparable specificity (93% and 96%, respectively). The positive predictive value and negative predictive value of the two-gene panel was 86% and 95%, respectively. CONCLUSIONS: Detection of the methylated TWIST1 and NID2 genes in urine sediments using MSP provides a highly (> or = 90%) sensitive and specific, noninvasive approach for detecting primary BCa. TRIAL REGISTRATION: BlCa-001 study - EudraCt 2006-003303-40.

TVT-O for the treatment of female stress urinary incontinence: results of a prospective study after a 3-year minimum follow-up.

OBJECTIVES: Medium-term results of transobturator tapes for the treatment of female stress urinary incontinence (SUI) are largely unknown. We analyzed the 3-yr results of a prospective, observational study designed to evaluate the safety and efficacy of the TVT-O procedure. METHODS: Preoperative and postoperative evaluations included physical examination, uroflowmetry and postvoid residual (PVR) measurement, and urinary symptoms and quality of life (QoL) questionnaires. Data were compared by means of the Wilcoxon matched pairs test. RESULTS: Between March 2003 and December 2003, 102 consecutive patients with clinical and urodynamic diagnoses of SUI who fulfilled inclusion and exclusion criteria underwent the TVT-O procedure; the latter was associated with pelvic organ prolapse treatment in 16 patients (15.7%). Three-year minimum follow-up (median, 40 mo) was available for 91 patients (89.2%). No erosion or persistent pain was noted. Four patients required tape release or section. Disappearance and improvement of SUI were observed in 88.4% and 9.3% of the patients, respectively. These cure rates were similar to those obtained 1 yr after the operation (p=0.55). Frequency and urge symptoms were improved at 3 yr (p<0.005). Whereas maximum flow rates were somewhat decreased (p=0.01), the severity of obstructive symptoms and PVR volumes were not statistically different (p=0.11 and p=0.32, respectively). Incontinence severity and QoL scale scores were largely better than preoperative ones (p<0.001) and did not differ from those reported at 1 yr (p=0.15 and p=0.08, respectively). CONCLUSIONS: The TVT-O procedure is a safe and efficient treatment of female SUI, with maintenance of high cure rates after a 3-yr minimum follow-up.