The oncogene Gankyrin is expressed in testicular cancer and contributes to cisplatin sensitivity in embryonal carcinoma cells.

BACKGROUND: Testicular germ cell cancer (TGCC) develops from pre-malignant germ neoplasia in situ (GCNIS) cells. GCNIS originates from fetal gonocytes (POU5F1+/MAGE-A4-), which fail to differentiate to pre-spermatogonia (POU5F1-/MAGE-A4+) and undergo malignant transformation. Gankyrin is an oncogene which has been shown to prevent POU5F1 degradation and specifically interact with MAGE-A4 in hepatocellular carcinoma (HCC) cells. We aimed to investigate the role of Gankyrin in progression from gonocyte to pre-invasive GCNIS and subsequent invasive TGCC. METHODS: We determined Gankyrin expression in human fetal testicular tissue (gestational weeks 9-20; n = 38), human adult testicular tissue with active spermatogenesis (n = 9), human testicular tissue with germ cell maturation delay (n = 4), testicular tissue from patients with pre-invasive GCNIS (n = 6), and invasive TGCC including seminoma (n = 6) and teratoma (n = 7). Functional analysis was performed in-vitro by siRNA knock-down of Gankyrin in the NTera2 cells (derived from embryonal carcinoma). RESULTS: Germ cell expression of Gankyrin was restricted to a sub-population of prespermatogonia in human fetal testes. Nuclear Gankyrin was also expressed in GCNIS cells of childhood and adult pre-invasive TGCC patients, and in GCNIS from seminoma and non-seminoma patients. Cytoplasmic expression was observed in seminoma tumour cells and NTera2 cells. Gankyrin knock-down in NTera2 cells resulted in an increase in apoptosis mediated via the TP53 pathway, whilst POU5F1 expression was unaffected. Furthermore, Gankyrin knock-down in NTera2 cells increased cisplatin sensitivity with an increase in cell death (13%, p < 0.05) following Gankyrin knock-down, when compared to cisplatin treatment alone, likely via BAX and FAS. Our results demonstrate that Gankyrin expression changes in germ cells during normal transition from gonocyte to prespermatogonia. In addition, changes in Gankyrin localisation are associated with progression of pre-invasive GCNIS to invasive TGCC. Furthermore, we found that Gankyrin is involved in the regulation of NTera2 cell survival and that a reduction in Gankyrin expression can modulate cisplatin sensitivity. CONCLUSIONS: These results suggest that manipulation of Gankyrin expression may reduce the cisplatin dose required for the treatment of TGCC, with benefits in reducing dose-dependent side effects of chemotherapy. Further studies are required in order to assess the effects of modulating Gankyrin on GCNIS/TGCC using in vivo models.

'Real-life experience': recurrence rate at 3 years with Hexvix® photodynamic diagnosis-assisted TURBT compared with good quality white light TURBT in new NMIBC-a prospective controlled study.

PURPOSE: To compare the recurrence rate at 3 years (RR-3y) for non-muscle invasive bladder cancer (NMIBC) between good quality (GQ) PDD-TURBT and GQWL-TURBT where PDD is used in routine practice for all new tumours. METHODS: All new, consecutive, NMIBC that received "good quality" criteria first TURBT across a university hospital service were prospectively recruited to this study over a 4-year period. Data were prospectively collected on all WL-TURBTs performed in 2007/8 and compared with PDD-TURBT from 2009/10. Only resection meeting strict "good quality criteria" were included from each cohort to control for resection quality, then cases were further matched 1:1 based on demographic and pathological criteria. The primary outcome was overall and risk group-specific recurrence rate at 3 years. RESULTS: Of 808 patients recruited, 345 had GQ-TURBT for NMIBC and were included. RR-3y was significantly less for GQ-PDD overall [RR-3y: GQ-PDD: 57/146 (39.0%), GQ-WL: 72/135 (53.3%) OR = 0.56 (0.35-0.90) p = 0.02] and on a 1:1 matched pair basis [RR GQ-PDD: 29/118 (24.6) vs. 59/118 (50.0) OR 0.33 (0.19-0.57) p < 0.001)]. Benefit was most marked in high-risk patients: RR-3y in high-risk patients treated with GQ-PDD was 25/48 (52.1%) vs. 28/35 (80%) for GQ-WL [OR 0.27 (0.10-0.74) p = 0.01]. CONCLUSION: When adopted for all new bladder tumour resections in routine practice, PDD appears to be associated with significantly reduced recurrence rates at 3 years in our "real life" experience, particularly in high-risk patients.

Intratubular germ cell neoplasia of the human testis: heterogeneous protein expression and relation to invasive potential.

Testicular germ cell cancer develops from premalignant intratubular germ cell neoplasia, unclassified cells that are believed to arise from failure of normal maturation of fetal germ cells from gonocytes (OCT4(+)/MAGEA4(-)) into pre-spermatogonia (OCT4(-)/MAGEA4(+)). Intratubular germ cell neoplasia cell subpopulations based on stage of germ cell differentiation have been described, however the importance of these subpopulations in terms of invasive potential has not been reported. We hypothesized that cells expressing an immature (OCT4(+)/MAGEA4(-)) germ cell profile would exhibit an increased proliferation rate compared with those with a mature profile (OCT4(+)/MAGEA4(+)). Therefore, we performed triple immunofluorescence and stereology to quantify the different intratubular germ cell neoplasia cell subpopulations, based on expression of germ cell (OCT4, PLAP, AP2γ, MAGEA4, VASA) and proliferation (Ki67) markers, in testis sections from patients with preinvasive disease, seminoma, and non-seminoma. We compared these subpopulations with normal human fetal testis and with seminoma cells. Heterogeneity of protein expression was demonstrated in intratubular germ cell neoplasia cells with respect to gonocyte and spermatogonial markers. It included an embryonic/fetal germ cell subpopulation lacking expression of the definitive intratubular germ cell neoplasia marker OCT4, that did not correspond to a physiological (fetal) germ cell subpopulation. OCT4(+)/MAGEA4(-) cells showed a significantly increased rate of proliferation compared with the OCT4(+)/MAGEA4(+) population (12.8 versus 3.4%, P<0.0001) irrespective of histological tumor type, reflected in the predominance of OCT4(+)/MAGEA4(-) cells in the invasive tumor component. Surprisingly, OCT4(+)/MAGEA4(-) cells in patients with preinvasive disease showed significantly higher proliferation compared to those with seminoma or non-seminoma (18.1 versus 10.2 versus 7.2%, P<0.05, respectively). In conclusion, this study has demonstrated that OCT4(+)/MAGEA4(-) cells are the most frequent and most proliferative cell population in tubules containing intratubular germ cell neoplasia, which appears to be an important factor in determining invasive potential of intratubular germ cell neoplasia to seminomas.

How painful is adult circumcision? A prospective, observational cohort study.

PURPOSE: Men are particularly concerned about pain after circumcision. Concerns about pain can be a reason to refuse surgery. We assessed the severity of postoperative pain and investigated factors that may influence postoperative pain. MATERIALS AND METHODS: We performed a prospective, observational cohort study in patients undergoing circumcision. Patients were asked to complete a questionnaire using a visual analog scale for pain (severity range 0 to 10) on days 1 to 3, 7 and 21, and record the analgesia used, complications and time off work. Other data recorded were patient age, clinical indication for surgery, foreskin retractility, presence of adhesions and histology. RESULTS: Of 211 questionnaires 112 were returned (53.1%). Mean patient age was 46.4 years. The most common clinical indication for circumcision was phimosis (75% of patients). Postoperative pain was scored as mild to moderate, including a mean of 2.4 on days 1 to 3, 2.1 on day 7 and 0.5 on day 21. Patients younger than 35 years (p = 0.025) and patients with wound infection (p = 0.036) had higher pain scores. Only 11 patients (9.8%) had severe pain at any time during recovery, including 8 with wound problems. Average ± SD time off work in the employed population was 6.6 ± 6.5 days, including 5 days for light work and 11 days for heavy physical activity. CONCLUSIONS: Pain is mild to moderate after circumcision in adults under general anesthesia with an intraoperative penile block. Severe pain is rare and mostly related to complications. Younger patients generally have more discomfort.

Establishing a reference range for penile length in Caucasian British men: a prospective study of 609 men.

OBJECTIVES: • To establish a reference range for adult male genital size in the UK using penile length measurements. • To compare the reference ranges for normal penile length reported from several different countries and the anthropometric differences noted between different nationalities and ethnic backgrounds. METHODS AND MATERIALS: • Over 20 months, genital measurements were taken from all men undergoing routine examination in clinics (n= 499) and in operating theatres during examination under anaesthetic (n= 110). • Using a rigid metric ruler three penile measurements were taken: flaccid pendulous penile length, flaccid penopubic penile length (to the pubic arch) and stretched flaccid penopubic length. In addition, testicular size was measured using an orchidometer. • The patient's age and the reason for referral were recorded. • Statistical analysis was carried out using Pearson correlation analysis. RESULTS: • Measurements from 610 patients aged 16-90 years were available for analysis. • The mean penile lengths were: pendulous length 8.7 cm (sd 1.6 cm), penopubic length 10.2 cm (sd 1.4 cm) and stretched length 14.3 cm (sd 1.7 cm). The mean testicular volume was 19.8 mL (sd 5.4 mL) for both left and right testicles. • Men with penile disease (including phimosis and Peyronie's disease) had slightly reduced penile length (pendulous -3.3 mm, P= 0.014; penopubic -2.3 mm, P= 0.029; stretched -5.1 mm, P < 0.001) compared with other referral groups (erectile dysfunction, testicular disease, prostate and bladder disease). • There was no significant correlation between penile length and age or testicular size CONCLUSION: • These data establish a reference range for adult male genital size in the UK, which should be helpful for urologists when counselling patients.

Expression of Intermediate Filaments in the Developing Testis and Testicular Germ Cell Cancer.

Cytokeratin and desmin expression have been associated with Sertoli cell maturity and the development of testicular germ cell cancer (TGCC). Thus, the present study aimed to characterize the expression of these intermediate filaments in normal testis development and TGCC. Cytokeratin and desmin were determined by immunohistochemistry and immunofluorescence in human fetal, and adult testis and tissue from patients with pre-invasive germ cell neoplasia in-situ (GCNIS) or invasive TGCC. Desmin was expressed in Sertoli cells of the human fetal testis, and the proportion of desmin expressing Sertoli cells was significantly reduced in the second trimester, compared with the first trimester (31.14% vs. 6.74%, p = 0.0016). Additionally, Desmin was expressed in the majority of Sertoli cells in the adult testis and TGCC samples. Cytokeratin was detected in Sertoli cells of human fetal testis but was not expressed in Sertoli cells of human adult testis. In patients with TGCC, cytokeratin was not expressed in Sertoli cells in tubules with active spermatogenesis but was detected in Sertoli cells in tubules containing GCNIS cells in patients with both pre-invasive and invasive TGCC. In conclusion, desmin was not associated with Sertoli cell maturation or progression to TGCC. However, cytokeratin appeared to be an indicator of impaired Sertoli cell maturation.

Evaluating the Safety of Performing Flexible Cystoscopy When Urinalysis Suggests Presence of "Infection": Results of a Prospective Clinical Study in 2350 patients.

BACKGROUND: There is significant underutilisation of allocated health service resources when a scheduled flexible cystoscopy (FC) is cancelled because a pre-cystoscopy urinalysis (PCU) suggests "infection", despite patients being asymptomatic for urinary tract infection (UTI). OBJECTIVE: To evaluate the risk of UTI or urinary sepsis when FC is performed in asymptomatic patients with a PCU positive for leucocyte esterase and/or nitrites. DESIGN SETTING AND PARTICIPANTS: A prospective cohort study was conducted in a high-volume UK centre recruiting all patients undergoing outpatient FC. INTERVENTION: A protocol was developed to guide response to PCU performed prior to FC, which was performed regardless of the result, unless patients were symptomatic for UTI. All patients completed a questionnaire to identify risk factors and were followed up via a telephone survey and a review of electronic clinical records. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Post-FC UTI was defined as hospital admission with UTI/urinary sepsis or if patients were symptomatic for UTI with receipt of antibiotics or with positive urine culture and sensitivity. An analysis of the association was performed. RESULTS AND LIMITATIONS: An initial pilot study confirmed the safety and feasibility of our protocol. Of 1996 patients, 136 (6.8%) developed a UTI by our definition, with 51 (2.6%) having a culture-proven infection. The risk was higher in patients with a positive PCU (odds ratio [OR] 1.61, 95% confidence interval [CI] = 1.07-2.40, p = 0.02), history of UTI (OR 1.72, 95% CI = 1.09-2.73, p = 0.02), or a bladder tumour on FC (OR 2.22, 95% CI = 1.27-3.90, p = 0.005). No patient with a positive PCU developed urinary sepsis. The main limitation of this study was the lack of pre-protocol control. CONCLUSIONS: We observed a clinically low and acceptable risk of UTI, with no incidence of sepsis, when FC was performed in asymptomatic patients with a PCU suggesting "infection". Routine cancellation of these patients is unnecessary and may worsen the burden on health service resources. PATIENT SUMMARY: We evaluated the safety of performing flexible cystoscopy when the urine dipstick on the day suggested presence of an "infection" but the patient had no symptoms of urinary tract infection (UTI). Our study in over 2000 patients demonstrated a low incidence of UTI, and none of these patients developed sepsis. We therefore recommend that flexible cystoscopy should not be cancelled automatically on the basis of the dipstick result alone, as it might delay a time-sensitive crucial diagnosis.

Screening for metabolic syndrome and testosterone deficiency in patients with erectile dysfunction: results from the first UK prospective study.

OBJECTIVE: To screen patients with erectile dysfunction (ED) for the presence of metabolic syndrome (MetS), testosterone deficiency and cardiovascular (CV) risk factors, in a secondary referral centre in the UK, as men with ED have a high incidence of CV risk factors that might amount to MetS, with obesity, increased risk of coronary heart disease and type II diabetes; testosterone deficiency has also been associated with both ED and MetS. PATIENTS AND METHODS: We assessed 124 men presenting with ED between March 2007 and August 2008. Data were collected prospectively for patient demographics, risk factors associated with MetS, and hypogonadism. MetS was assessed using the National Cholesterol Education Program Adult Treatment Panel III Criteria 2005 (based on three or more of five criteria: waist circumference, high triglycerides, low levels of high-density lipoprotein cholesterol, hypertension and impaired glucose tolerance). RESULTS: The mean (range) age of the men was 50 (16-76) years; 50 of 124 (40%) patients had MetS and 27% had hypogonadism. The latter was also associated with a low testicular volume and decreased libido. Ninety-seven patients (82%) were either overweight or obese, and 64 (52%) were current or ex-smokers. CONCLUSIONS: Our audit confirms a high incidence of MetS and hypogonadism in patients with ED in the UK. We recommend routine screening for CV risk factors, MetS and testosterone deficiency in all patients in the UK with ED.

Testicular cancer: improving outcomes with national quality performance indicators.

Testicular cancer is the most common malignancy in young adult men. The prognosis is excellent in limited disease and cure is possible even in advanced disease. Quality performance indicators (QPI) are used in many developed countries as a measure of healthcare performance. We report and discuss the development of a national set of QPIs in Scotland for testicular cancer as a method of gathering demographic data and driving improvement in nationwide testicular cancer outcomes.

Predicting Grade and Stage at Cystoscopy in Newly Presenting Bladder Cancers-a Prospective Double-Blind Clinical Study.

OBJECTIVE: To assess urologists' ability to predict the grade and stage of new bladder cancers from the cystoscopic features alone. PATIENTS AND METHODS: We conducted a prospective clinical study on consecutive patients who underwent transurethral resection of bladder tumor (TURBT) for new bladder cancers. Using only cystoscopic tumor morphology at the time of initial TURBT, 3 urology consultants predicted the grade and stage, recording these on a proforma along with tumor features. Predictions were compared with assessments by uropathologists, blinded to the clinical prediction. We analyzed the accuracy in (1) predicting the exact grade and stage of the cancer; (2) differentiating between low- and high-grade cancers; and (3) discerning between Ta, T1, and T2 cancers. RESULTS: Of 248 patients, 224 were suitable for analysis. The positive predictive values for low- and high-grade cancers were 85.8% and 71.3%, respectively. The overall likelihood of a consultant predicting high-grade cancers as being low grade was 16/83 (19.3%). When tumors were large (>30 mm), this likelihood dropped significantly to 7.3% (4/55) (odds ratio = 3.1, 95% confidence interval = 1.0-9.7, P = .04). Non-muscle-invasive and muscle-invasive cancers were predicted accurately in 93.4% and 85.2% patients, respectively. Six of 161 (3.7%) tumors predicted to be non-muscle-invasive bladder cancer were actually muscle invasive on histology. CONCLUSION: For clinical purposes, in newly presenting patients with bladder cancer, urologists appear to reliably predict lower grade and muscle-invasive disease, confirming widely held belief. This allows for appropriate and efficient use of surgical expertise, available technology, and selection of participants for clinical trials on the basis of prehistology risk categories.

Experimentally induced testicular dysgenesis syndrome originates in the masculinization programming window.

The testicular dysgenesis syndrome (TDS) hypothesis, which proposes that common reproductive disorders of newborn and adult human males may have a common fetal origin, is largely untested. We tested this hypothesis using a rat model involving gestational exposure to dibutyl phthalate (DBP), which suppresses testosterone production by the fetal testis. We evaluated if induction of TDS via testosterone suppression is restricted to the "masculinization programming window" (MPW), as indicated by reduction in anogenital distance (AGD). We show that DBP suppresses fetal testosterone equally during and after the MPW, but only DBP exposure in the MPW causes reduced AGD, focal testicular dysgenesis, and TDS disorders (cryptorchidism, hypospadias, reduced adult testis size, and compensated adult Leydig cell failure). Focal testicular dysgenesis, reduced size of adult male reproductive organs, and TDS disorders and their severity were all strongly associated with reduced AGD. We related our findings to human TDS cases by demonstrating similar focal dysgenetic changes in testes of men with preinvasive germ cell neoplasia (GCNIS) and in testes of DBP-MPW animals. If our results are translatable to humans, they suggest that identification of potential causes of human TDS disorders should focus on exposures during a human MPW equivalent, especially if negatively associated with offspring AGD.

Real-life Experience: Early Recurrence With Hexvix Photodynamic Diagnosis-assisted Transurethral Resection of Bladder Tumour vs Good-quality White Light TURBT in New Non-muscle-invasive Bladder Cancer.

OBJECTIVE: To compare early recurrence between good-quality white-light transurethral resection of bladder tumor (GQ-WLTURBT) and photodynamic diagnosis-assisted (PDD) transurethral resection of bladder tumor (TURBT) in a real-life controlled setting. METHODS: A prospective controlled study was conducted commencing with a planned prospective cohort of patients with new tumors undergoing white-light TURBT in 2007-2008. Previously defined principles of GQ-WLTURBT for standardization and comparison of TURBT techniques, which are (1) cystoscopic mapping using a bladder diagram, (2) documented complete resection of the tumor, (3) resection performed or supervised by an experienced surgeon, (4) presence of detrusor muscle in the specimen, and (5) patient receiving mitomycin C within 24 hours of the resection, were applied. This was followed by a prospective cohort of new patients undergoing PDD-TURBT in 2009-2011. Only patients with new non-muscle-invasive bladder cancer (NMIBC) deemed to have had complete first TURBT were included for analysis. Tumor features and findings at first check cystoscopy and early re-TURBT (in high-risk NMIBC) were evaluated. Early recurrence (for calculating recurrence rate at first follow-up cystoscopy) was defined as pathologically confirmed tumor on early re-TURBT or recurrence at the first check cystoscopy. Comparison was analyzed between GQ-WLTURBT and good-quality PDD-TURBT (GQ-PDDTURBT). RESULTS: A total of 808 patients were evaluated. The overall RRFFCs for GQ-WLTURBT and GQ-PDDTURBT were 30.9% and 13.6%, respectively (odds ratio = 2.9; 95% CI = 1.6-5.0; P <.001), with statistically significant lower recurrence rates in low- and intermediate-risk NMIBC after GQ-PDDTURBT. CONCLUSION: Hexvix PDD-assisted TURBT is associated with a significantly lower risk of early recurrence compared with GQ-WLTURBT in a real-life clinical setting.

Adult prepuceplasty: comparison of outcomes of standard prepuceplasty and foreskin Z-plasty.

OBJECTIVE: To compare the clinical and patient-reported outcomes from standard prepuceplasty and foreskin Z-plasty. METHODS: Consecutive standard prepuceplasty (n = 22) and Z-plasty (n = 12) procedures performed from September 2005 to December 2010 were analyzed. The patient and operative data were collated, together with the results of a patient questionnaire inquiring about the postoperative complications, postoperative pain, remaining foreskin tightness in the flaccid or erect penis, cosmetic appearance after surgery, and the need for later circumcision. RESULTS: The median follow-up period was 26 months for standard prepuceplasty and 16 months for Z-plasty. Of the 22 patients in the standard prepuceplasty group and 12 patients in the Z-plasty group, 5 (22.7%) and 1 (9.1%) requested circumcision (P = .33), respectively. One man in each group underwent revision standard prepuceplasty. Patients undergoing the 2 procedures reported no significant differences for the questions relating to postoperative pain and foreskin tightness in the flaccid or erect penis. However, the cosmetic appearance after Z-plasty, assessed using a Likert scale question, were "good" or "very good" compared with "acceptable" for the standard group (P = .005). CONCLUSION: Prepuceplasty can be used successfully in well-selected patients. In the present series, 82% of men avoided circumcision. Z-plasty resulted in fewer circumcisions and a significantly better cosmetic appearance than standard prepuceplasty.

Improved detection of urothelial carcinoma in situ with hexaminolevulinate fluorescence cystoscopy.

PURPOSE: In this European multicenter study we compared hexaminolevulinate (HAL) fluorescence cystoscopy and standard white light cystoscopy for the detection of carcinoma in situ (CIS) in patients suspected of having high risk bladder cancer. MATERIALS AND METHODS: This study was a prospective controlled, within-patient comparison of standard and HAL fluorescence cystoscopy. Eligible patients received an intravesical instillation of 50 ml HAL 8 mM solution. Cystoscopy was performed using a D light system, which provided white and blue light at 375 to 440 nm. The bladder wall was inspected and mapped, first under white light, followed by blue light. All tumors and suspicious areas identified under white light and by red fluorescence were resected or biopsied. Histological findings were assessed by an independent central pathologist blinded to the identity of the biopsies. RESULTS: Of 211 evaluable patients 83 (39%) had CIS, of whom 18 (22%) were detected by HAL cystoscopy only, 62 (75%) were detected by standard and HAL cystoscopy, 2 (2%) were detected by standard cystoscopy only and 1 (1%) was detected by nonguided biopsy. Therefore, HAL cystoscopy identified 28% more patients with CIS than standard cystoscopy. The side effects of HAL instillation were negligible and no unexpected events were reported. CONCLUSIONS: HAL fluorescence cystoscopy improves the detection of bladder CIS significantly, which has consequences for clinical management and may improve the patient prognosis. The procedure is easily implemented as an adjunct to standard cystoscopy and it adds no significant risk of complications.