[Malignant Sertoli cell tumors of the testes]. / Maligner Sertoli-Zell-Tumor des Hodens.

Sertoli cell tumors are a rare malignancy which account for approximately 1.5 % of all testicular tumors. Although malignant Sertoli cell tumors are uncommon, they are associated with a poor prognosis. So far 36 cases of malignant courses of disease have been described. We present a patient with a lymphogenic metastasized Sertoli cell tumor, who 24 months after orchiectomy and extended retroperitoneal lymphadenectomy is relapse-free.

Evaluation of perioperative complications in open and laparoscopic surgery for renal cell cancer with tumor thrombus involvement using the Clavien-Dindo classification.

OBJECTIVE: To identify risk factors for perioperative complications and morbidity in renal cell cancer (RCC) surgery with tumor thrombus invasion (TTI). PATIENTS AND METHODS: Retrospective single-center analysis of 128 patients who underwent open (n = 97) or laparoscopic (n = 31) radical nephrectomy (NT) for RCC between 1999 and 2010. TTI was at Mayo-Level 0, I, II, III, IV in 88, 7, 10, 4, and 19 cases, respectively. Cavotomy was performed in 27, liver mobilisation in 20, and cardiovascular bypass in 17 patients. RESULTS: The rate of any early postoperative complication (PC) by Clavien-Dindo classification was 58.6%, while the severe early PC rate was 29.7%. There was a statistically significant difference in multivariate analysis in the incidence of any early PC and of severe early PC by Charlson score (OR:1.584 (95%CI:1.141-2.199), p = 0.006; OR:3.065 (95%CI:1.218-7.714), p = 0.017) and by tumor thrombus level TNM-UICC 2010 T3a/T3c (OR:10.668 (95%CI:1.266-89.871), p = 0.029; OR:10.502 (95%CI:2.981-36.992), p < 0.001). In pT3a cases open NT was associated with a higher early (57.9% vs. 25.8%) and severe (24.6% vs. 9.7%) PC rate compared to laparoscopic NT. The 30-day mortality rate was 0%. The 90-day mortality rate was 6.3% but 100% cancer-related. In Cox regression analysis tumor thrombus level was not predictive for overall survival. CONCLUSIONS: The strongest risk factor for early and severe PC in patients with TTI is a supradiaphragmatic tumor thrombus. In cases with severe PC, this fact persists when comparing Mayo-Levels II-III and Level IV. In pT3a cases open NT shows a 2-fold higher early PC rate compared to laparoscopic NT.

[Propensity score comparison of the various radical surgical techniques for high-risk prostate cancer]. / Propensity Score Vergleich der verschiedenen radikalen Operationstechniken beim high risk Prostatakarzinom.

INTRODUCTION: The optimal surgical treatment of patients with a high risk prostate cancer (PCa) in terms of radical prostatectomy (RP) is still controversial: open retropubic RP (RRP), laparoscopic RP (LRP), or robot-assisted (RARP). We aimed to investigate the influence of the different surgical techniques on pathologic outcome and biochemical recurrence. PATIENTS AND METHODS: A total of 805 patients with a high risk PCa (PSA >20 ng/mL, Gleason Score ≥8, or clinical stage ≥cT2c) were included. A comparison of 407 RRP patients with 398 minimally invasive cases (LRP+RARP) revealed significant confounders. Therefore all 110 RARP cases were propensity score (PS) matched 1:1 with LRP and RRP patients. PS included age, clinical stage, preoperative PSA, biopsy Gleason score, surgeon's experience and application of a nerve sparing technique. Comparison of overall survival (OS) and recurrence-free survival (RFS) was done with the log rank test. Predictors of RFS were analyzed by means of Cox regression models. RESULTS: Within the post-matching cohort of 330 patients a pathologic Gleason score < 7, = 7 and > 7 was found in 1.8, 55.5 and 42.7% for RARP, in 8.2, 36.4, 55.5% for LRP and in 0, 60.9 and 39.1% for RRP (p=0.004 for RARP vs. LRP and p=0.398 for RARP vs. RRP). Differences in histopathologic stages were not statistically significant. The overall positive surgical margin rate (PSM) as well as PSM for ≥ pT3 were not different. PSM among patients with pT2 was found in 15.7, 14.0 and 20.0% for RARP, LRP and RRP (statistically not significant). The respective mean 3-year RFS rates were 41.4, 77.9, 54.1% (p<0.0001 for RARP vs. LRP and p=0.686 for RARP vs. RRP). The mean 3-year OS was calculated as 95.4, 98.1 and 100% respectively (statistically not significant). CONCLUSION: RARP for patients with a high risk PCa reveals similar pathologic and oncologic outcomes compared with LRP and RRP.

Optimizing evaluation of split renal function in a living kidney donor using scintigraphy and calculation of the geometric mean: a case report.

Within the evaluation process of living kidney donors, split renal function is usually evaluated by renal scintigraphy. Since split renal function measured by conventional posterior scans depends on the position of the kidney, actual suitable donors may be rejected because of an inaccurate examination technique. We report the case of a 28-year-old male living kidney donor. Due to a complex vascular anatomy of the right kidney, only his left kidney was considered eligible for transplantation. In conventional posterior Tc99m-mercapto-acetyltriglycine scintigraphy, the left kidney had a relative function of 60%. A second scintigraphy using anterior and posterior dimercaptosuccinic acid scans with calculation of the geometric mean showed an adapted relative function of the left kidney of 53%, now meeting the inclusion criteria for living kidney donation. This case shows that the geometric mean method using simultaneous anterior and posterior views obtained with a dual-head gamma camera can be a very helpful approach to determine split renal function of potential living kidney donors. Further investigation is necessary to prove the benefit of a general bilateral scan before living kidney donation.

Outcome of expanded criteria donor kidneys that were transplanted at other Eurotransplant centers after being rejected by our institution.

PURPOSE: There is a growing discrepancy between the demand for renal transplants and the number of transplants conducted. For the many patients on the renal transplant waiting list, this means increased dialysis-associated morbidity, mortality and a reduced quality of life. The aim of this study was to ascertain whether it is justifiable for transplant centers to reject cadaveric donor organs on hand of marginal organ quality. METHODS: We identified 110 kidneys that were primarily rejected for transplantation at Charité Universitätsmedizin Berlin, Campus Mitte, and later transplanted at another center within the Eurotransplant zone. Using data from the Collaborative Transplant Study, we analyzed various demographic donor data including cold ischemia times, as well as graft and recipient outcomes. RESULTS: The median follow-up was 54 months. The cold ischemia time averaged 16 h. The organs that were primarily rejected by our center and then transplanted at other Eurotransplant centers showed 31 % of recipients had creatinine levels under 1.47 mg/dl and 94 % had levels under 2.97 mg/dl at 3-year follow-up. The mean death-censored graft survival was 71.4 months. The mean renal transplant recipient survival was 87.5 months. CONCLUSIONS: Based on our findings, we propose that acceptance criteria for marginal donor kidneys need to be widened.

[Introduction of interdisciplinary prostate cancer centers based on the recommendations of the German Cancer Society. A cost-benefit analysis 3 years after accreditation]. / Etablierung von interdisziplinären Prostatakrebszentren nach den Empfehlungen der DKG. Eine Kosten-Nutzen-Analyse 3 Jahre nach Zertifizierung.

The introduction of prostate cancer treatment centers according to the criteria of the German Cancer Society ("Deutsche Krebsgesellschaft", DKG) aims at improving the quality of care for patients with prostate cancer. Systematic analyses of the effects and costs are lacking as yet. Three years after certification of the Interdisciplinary Prostate Cancer Center at the Charité Hospital Berlin we observed a decrease in the rate of positive surgical margins (tumor stage pT2), but other parameters of treatment quality including patient satisfaction remained unchanged. A survey among urologists of the region showed a high acceptance of prostate cancer centers in general. The majority of participating urologists appreciated the work of the Charité center, in particular the treatment recommendations given by the center were mostly followed and the majority of urologists regularly use educational activities of the center. However, only 30% of the participating urologists confirmed short-term improvements in the quality of patient care. Yearly additional costs for the Charité prostate cancer center are estimated at 205,000 euro (precertification phase and certification) and 138,000 euro (monitoring phase), despite the initial drop in mean treatment costs per case (radical prostatectomy). The introduction of prostate cancer treatment centers certified by the DKG is cost intensive, increases in treatment efficiency notwithstanding. Short-term improvements in quality of care cannot be unequivocally demonstrated. Prostate cancer centers serve an important role in counseling and medical education and may thus help disseminate evidence-based treatment strategies.

[Evaluation of obstructive interval, presence of sperm granulomas and patient age as predictors of spermatogenesis in a cohort of men undergoing vasectomy reversal]. / Evaluation von Vasektomieintervall, Spermagranulom und Patientenalter als Prädiktoren der spermatogenetischen Aktivität vasektomierter Männer.

INTRODUCTION: For more than 50 years vasectomy reversal is a routinely performed procedure in the field of urology. However, there is no scientific agreement about morphological changes of the testes caused by vasectomy. The existing evidence for reduced fertility rates following vasectomy reversal demands a clear statement regarding potential histological changes and impaired spermatogenesis following vasectomy. Thus far there is little knowledge about potential histological changes of the testis caused by vasectomy. MATERIAL AND METHODS: 330 consecutive patients who underwent vasectomy reversal had bilateral testicular biopsies which were evaluated utilising semi-thin sections. The number of mature spermatids per seminiferous tubule was considered the variable of interest as it represents an objective and reproducible parameter of spermatogenesis. The number of mature spermatids per tubule was correlated with patient age, obstructive interval and the presence or absence of sperm granulomas via the chi-square test. RESULTS: Overall, 570 sections of 285 patients were eligible for evaluation. The mean patient age was 41.2 years (range 27-63 years, SD +/- 6.5 years) with a mean obstructive interval of 105.9 months (range 12-328, SD +/- 66.1). 56 patients (19.6 %) had a sperm granuloma on the right and 22 (11.6 %) on the left ductus deferens. There was no statistically significant correlation between the presence of a sperm granuloma with the number of mature spermatids per tubule (p = 0.717). Furthermore, there was neither an association of obstructive interval (p = 0.144) nor patient age (p = 0.168) with spermatogenesis. CONCLUSION: Regular spermatogenetic activity in all examined samples with development of mature spermatids was shown. Furthermore, for the first time we were able to demonstrate in a large cohort of patients that neither patient age nor obstructive interval nor sperm granuloma have a significant impact on spermatogenesis.

[What is most important is what comes across : Urological guidelines from the target group's point of view]. / Entscheidend ist, was ankommt : Urologische Leitlinien aus Sicht der Zielgruppe.

BACKGROUND: Guidelines are developed to improve the quality of patient care. The effect of German urologic guidelines has not been evaluated so far. Therefore, we aimed to systematically investigate the acceptance, use, and quality of the published guidelines from a user's perspective. METHODS: A link to an online questionnaire concerning use and barriers to the application of guidelines was distributed via e-mail by the German Society of Urology (DGU). German urologists' opinions on differences in national guideline quality were evaluated regarding prostate cancer (PCA), bladder cancer, germ cell tumors (GCT), renal cell carcinomas, and erectile dysfunction. RESULTS: Four hundred sixty-seven German urologists participated. More than 90% of the participants considered guidelines to be helpful. The Internet as the main tool for guideline distribution was favored by 28.4%, followed by publication in Urologe A. The main barrier to guideline usage was attributed to the lack of up-to date clinical data. Guidelines for GCT scored best in all quality categories and reached the highest level of use (65.8%), and 40.5% of participating urologists considered the additional establishment of comprehensive care centers for GCT as more effective for quality improvement than guideline development alone. For the other urologic tumors, especially PCA, guideline development was favored as a tool for quality improvement. CONCLUSION: More than 90% of participating urologists accept clinical guidelines as useful instruments in clinical practice and for therapeutic decisions. Our results should be integrated into guideline dissemination and implementation strategies in order to achieve a higher degree of treatment conformation to guidelines.

Impact of obstructive interval and sperm granuloma on patency and pregnancy after vasectomy reversal.

The objective of this study was to determine the effect of the obstructive interval and the presence of a sperm granuloma on vas patency and pregnancy rate following vasectomy reversal. We identified 334 patients with complete follow-up who met the inclusion criteria. There were significant associations between the obstructive interval and procedure performed as well as with patient age. Patients with longer obstructive intervals were more often older (p < 0.001) and more likely to have a vaseoepididymostomy performed (p < 0.001). There was no association between the presence of a sperm granuloma or the length of the obstructive interval with post-operative vas patency and pregnancy rates. The only independent predictor of post-operative fertility was age of the female partner (p = 0.015). Our data clearly demonstrates that when state of the art surgical techniques are used, neither the presence of a sperm granuloma nor the obstructive interval serve as prognosticators of post-operative vas patency and pregnancy rates. However, when counselling patients and their female partners, it is of utmost importance to stress that the age of the female partner is an independent predictor of successful vasectomy reversal.

[Testicular cancer - explanatory models for high cisplatin chemosensibility and new therapeutic options]. / Testikuläre Keimzelltumoren - Erklärungsmodelle für die hohe Ansprechrate auf Cisplatin-basierte Chemotherapien und neue Therapieoptionen.

This article offers a review about the current facts of chemotherapy in testicular cancer. Besides a short presentation of the guideline-standard therapy the authors deal with the question as to why testicular cancer shows an extraordinarily high chemosensibility compared to other tumours. Furthermore, the current data on alternative chemotherapies as well as of molecular, molecular-genetic and pharmacogenetic therapeutic concepts are explored. Data were obtained from researches in Medline of the Pubmed database.

[Expression levels of the IAP antagonists XAF1, Smac/DIABLO and HtrA2 in testicular germ cell tumours]. / Expression der pro-apoptotischen Apoptoseinhibitor-(IAP)Antagonisten XAF1, Smac/DIABLO und HtrA2 in Keimzelltumoren des Hodens.

PURPOSE: Down-regulation of the IAP antagonistis XAF1, Smac/DIABLO and HtrA2, has been related to the onset and progression of various malignancies. We examined the mRNA-expression of these pro-apoptotic parameters in testicular germ cell tumors (TGCT) and normal testicular tissue and correlated their expression levels to clinicopathological tumour features. MATERIAL AND METHODS: Real-time RT-PCR was used to quantify the mRNA-expression of XAF1, Smac/DIABLO and HtrA2 in normal testicular tissue (n = 18), carcinoma in situ (n = 4), seminomas (n = 64), and non-seminomatous germ cell tumors (n = 35). RESULTS: Compared to normal testicular tissue, the expression levels of XAF1 were increased in TGCT (p < 0.001), whereas those of Smac/DIABLO and HtrA2 were decreased (p < 0.001 and p < 0.001). Smac/DIABLO expression levels showed a significant trend towards a gradual decrease from normal testicular tissue to CIS and seminomas and finally to NSGCT (p < 0.001). Moreover, XAF1 and HtrA2 expression levels gradually increased with progression of clinical tumour stage in seminoma patients (p = 0.001 and p = 0.018), their expression levels being strongly intercorrelated (Spearman rho correlation coefficient: 0.674; p < 0.001). CONCLUSION: These data suggest that a down-regulation of Smac/DIABLO and HtrA2 is implicated in the development and progression of TGCT, whereas overexpression of XAF1 in TGCT might contribute to their extraordinary sensitivity to chemotherapy. Regarding the additional correlation of XAF1 and HtrA2 expression with clinical tumour stage in seminoma patients, it appears reasonable to further evaluate these three IAP antagonists as molecular parameters for the prediction of treatment response and prognosis of TGCT patients.

Expression parameters of the polycomb group proteins BMI1, SUZ12, RING1 and CBX7 in urothelial carcinoma of the bladder and their prognostic relevance.

BACKGROUND: The polycomb group (PCG) proteins are epigenetic transcriptional repressors involved in the control of cellular proliferation and oncogenesis. This study aimed at examining whether mRNA tumor levels of the PCG family members BMI1, SUZ12, RING1, and CBX7 relate to histopathological parameters in urothelial carcinomas of the bladder and whether they may provide prognostic information following tumor resection. METHODS: The relative gene expression of BMI1, SUZ12, RING1, and CBX7 was analyzed by real-time RT-PCR in tumor tissue obtained from 93 patients with urothelial carcinoma of the bladder undergoing surgical treatment. Expression data was correlated with pathological variables and outcome. RESULTS: PCG family members BMI1, SUZ12, RING1, and CBX7 are commonly expressed in urothelial carcinomas of the bladder. The relative CBX7 mRNA expression levels gradually decreased from superficial (pTa) to invasive (pT1) and finally to muscle-invasive (> or =pT2) tumors (p = 0.008). Furthermore, CBX7 expression was statistically significantly correlated with tumor grade (p = 0.04). No correlation of mRNA levels with histopathological tumor features or tumor recurrence was observed for the other PCG components investigated. CONCLUSION: Expression levels of CBX7 inversely correlate with the progression of tumor stage and grade in urothelial carcinomas of the bladder, suggesting that downregulation of CBX7 indicates aggressive urothelial carcinoma phenotype.

[New polymorphisms in the PSA promoter in patients with prostate carcinoma]. / Neue polymorphismen im PSA-promoter bei prostatakrebs.

INTRODUCTION: PSA is still the most important parameter in the diagnosis and follow-up of prostate cancer. We searched for single nucleotide polymorphisms (SNP) in four different parts of the PSA promoter, which harbour binding sites for major transcriptional regulators using PCR-based combined SSCP and sequence analysis. MATERIALS AND METHODS: Lymphocyte DNA samples from 279 prostate cancer patients and 55 age-matched controls were subjected to SSCP analysis after PCR amplification of four approximately 200-bp fragments selected to contain the known AREs I-III or the transcriptional start site, respectively. Conspicuous PCR fragments with variant SSCP patterns were subsequently cloned and sequenced. Computer-assisted comparison with published sequences of the promoter region was performed to reveal polymorphic sites. RESULTS: In 66.6 % of the carcinoma cases DNA displayed polymorphisms in ARE I-, whereas the benign cases showed this SNP only in 14.5 %. This difference was highly significant (p < 0.0001). In addition, we found novel SNPs with lower frequency at positions - 179, - 230, - 233 (ARE I) and - 356 (ARE II). CONCLUSION: The present study confirmed that the otherwise already described polymorphism in the position - 158 is highly significantly more frequent in prostate cancer patients than in the control group. There is no significant correlation to the clinical stage of the disease. Furthermore we described for the first time four rare, previously unknown polymorphisms.

mRNA expression profiles of methylated APAF-1 and DAPK-1 tumor suppressor genes uncover clear cell renal cell carcinomas with aggressive phenotype.

PURPOSE: We determined the relation between promoter methylation and mRNA expression of the p53 target genes APAF-1 and DAPK-1 in 55 consecutive tumor and corresponding normal tissue samples. MATERIALS AND METHODS: Tissue was taken from nonmetastatic clear cell renal cell carcinoma at a median followup of 75 months. Quantitative methylation specific real-time polymerase chain reaction and quantitative real-time reverse transcriptase-polymerase chain reaction were used. RESULTS: The mRNA expression levels of APAF-1 and DAPK-1 were significantly higher in tumor vs nontumor tissue from the same kidney (p = 0.003 and 0.0001, respectively). Expression levels of the 2 genes did not correlate with tumor stage but they were significantly lower in high grade (G3) tumors (p = 0.018 and 0.05, respectively). In patients with positive lymph node staging mRNA expression of APAF-1 and DAPK-1 was significantly lower. On matched pair analysis of tumor tissue the methylation level of the APAF-1 gene correlated inversely with the mRNA expression level (p = 0.02). In tumor and normal kidney tissue from patients with lesions 4 cm or greater mRNA expression levels of DAPK-1 were significantly lower in those who later had metastatic disease (p = 0.04 and 0.02, respectively). CONCLUSIONS: These data demonstrate decreased tumor suppressor gene expression in more aggressive subtypes of clear cell renal cell carcinoma. The lower mRNA level of the DAPK-1 gene in tumor and normal tissue from patients with an unfavorable clinical outcome suggests the organ specific loss of tumor suppressor gene expression, predisposing to metastatic tumor disease and shorter overall survival.

Long-term efficacy of tolterodine and patient compliance in pediatric patients with neurogenic detrusor overactivity.

INTRODUCTION: We investigated the effects of standard oral anticholinergic treatment with tolterodine in children with neurogenic bladder over a 5-year follow-up period and focused on treatment satisfaction, patient compliance and urodynamic parameters. MATERIAL AND METHODS: The follow-up consisted of regular visits and urodynamic evaluation at least once a year. The patients or their parents were interviewed to evaluate voiding behavior, as well as factors leading to lower patient compliance and deterioration in urodynamic parameters. RESULTS: Of the 43 patients evaluated, 30 (70%) took their anticholinergic medication consistently and 13 (30%) sporadically. The mean bladder capacity was 354.7 ml in the first group but only 214.7 ml in the noncompliant group (p < 0.001). The mean maximal detrusor pressure decreased from 42.2 to 33.6 cm H(2)O in the compliant group (p < 0.001) and from 49.7 to 46.4 cm H(2)O in the noncompliant group (p = 0.21). The mean detrusor compliance increased from 18.9 to 19.3 ml/cm H(2)O in the compliant group (p = 0.63) and from 11.8 to 12.3 ml/cm H(2)O in the noncompliant group (p = 0.87). Side effects such as dry mouth (11/13) and dizziness (7/13) were common in the noncompliant group, whereas only 5/30 reported dry mouth in the compliant group. CONCLUSIONS: These data demonstrate the efficacy and tolerability of tolterodine over a long follow-up period. The results are promising in view of the fact that the patients will probably require life-long medication. Nevertheless, anticholinergic side effects still cause some patients to refuse regular medication, which results in a poorer urodynamic outcome.

EZH2 polycomb transcriptional repressor expression correlates with methylation of the APAF-1 gene in superficial transitional cell carcinoma of the bladder.

The EZH2 gene controls methylation of various EZH2 target promoters. The APAF-1, DAPK-1 und IGFBP-3 genes are frequently methylated in bladder cancer, and methylation of these genes is found in more aggressive tumor types. The aim of our study was to investigate a potential link between EZH2 mRNA expression and the extent of APAF-1, DAPK-1 and IGFBP-3 methylation in urothelial transitional cell carcinoma (TCC) and to correlate the data with histopathological parameters and follow-up data. EZH2 mRNA expression was measured by real-time reverse transcription polymerase chain reaction, and the methylation analysis was performed using methylation-specific real-time polymerase chain reaction. Tissue specimens were obtained from 35 patients with TCC. EZH2 mRNA expression was detected in all tumor specimens investigated. The EZH2 expression levels correlated well with the differentiation grade of the tumor specimens (p = 0.03), and the APAF-1 methylation correlated with tumor stage (p = 0.0001) and grade (p = 0.004). Matched pair analysis demonstrated a statistically significant correlation between elevated EZH2 mRNA expression and higher methylation levels of APAF-1 in superficial (p = 0.024) and well- differentiated (p = 0.04) TCC. In patients with recurrent TCC, APAF-1 and IGFBP-3 methylation levels were significantly higher (p = 0.03 and p = 0.01, respectively), which was not observed when EZH2 mRNA expression or DAPK-1 methylation levels were related to the clinical outcome. In conclusion, our data show that EZH2 expression and APAF-1 methylation are related to tumor progression and invasiveness. Moreover, these data present first evidence that APAF-1 methylation is related to transcriptional activity of EZH2 expression in early-stage tumor disease of the bladder.

Frequent epigenetic inactivation of p53 target genes in seminomatous and nonseminomatous germ cell tumors.

Hypermethylation of tumor-suppressor genes has been implicated in the pathogenesis of human cancers. This study was designed to examine the methylation profiles of a selected group of p53 target genes (APAF-1, CASP-8, DAPK-1, IGFBP-3) and to correlate the findings with the histopathological characterization of testicular germ cell tumors (TGCT). Promoter methylation status was analysed by highly sensitive real-time methylation-specific PCR in 46 primary TGCTs (26 seminomas and 20 nonseminomas) and 15 normal testicular tissue samples. APAF-1 methylation was detected in all of the seminomatous and nonseminomatous TGCTs as well as in 60% of normal testicular tissue. Methylation of DAPK-1 was frequent in seminomas (50%) and nonseminomas (20%), but not in normal testicular tissue (6%). The degree of DAPK-1 methylation correlated with the clinical stage of the disease (P=0.05) and was useful in differentiating seminomatous from nonseminomatous, and malignant from nonmalignant testicular tissue (P=0.04 and 0.02, respectively). The APAF-1 methylation index achieved a highly significant differentiation between seminomatous or nonseminomatous tissue and nonmalignant testicular tissue (P=0.0001). In testicular tumorigenesis, promoter methylation of specific p53 target genes occurs at early stage but to varying degrees. Methylation also occurs in normal testicular tissue, which is in contrast to findings in other urogenital malignancies. Further studies will be necessary to determine whether the methylation level may be used as marker for risk estimation, especially in clinical stage I disease.

Methylation of tumour suppressor genes APAF-1 and DAPK-1 and in vitro effects of demethylating agents in bladder and kidney cancer.

To examine the significance of the methylation level of the p53 target and tumour suppressor genes apoptotic protease activating factor-1 (APAF-1) and death-associated protein kinase-1 (DAPK-1) in 80 microdissected tumour samples from transitional cell carcinoma (TCC) of the bladder and 80 tumour samples from clear-cell renal cell carcinoma (RCC) as well as from non-tumourous bladder and kidney tissue. Growth-inhibitory effects of the demethylating agents 5-Aza-2'-deoxycytidine (5-Aza-CdR) and zebularine were investigated in TCC and RCC cell lines. The methylation frequency of APAF-1 (DAPK-1) was 100% (77%) in TCC and 100% (33%) in RCC. The methylation levels of APAF-1 could differentiate between the individual tumour stages in TCC as well as in RCC. The APAF-1 methylation levels in RCC were significantly higher in tumours larger than 4 cm and in high-grade tumours. The methylation frequencies in normal tissue for APAF-1 (DAPK-1) were 11% (8%) in bladder tissue and 9% (5%) in kidney tissue. The growth-inhibitory effect of the demethylating agents in TCC (RT4, T24) and RCC (A498, ClearCa-5) cell lines resulted in a 17-132% prolongation of the doubling time (DT). In RCC cell lines, zebularine was superior to 5-Aza-CdR in achieving a DT prolongation. Quantitative real time RT-PCR detected a re-expression of mRNA transcripts of APAF-1 or DAPK-1. In conclusion, demethylating agents effectively retard growth of TCC and RCC cell lines. Methylation level analysis of specific genes has the potential for further tumour characterisation in TCC and RCC.

[Rectal perforation as a late complication of ProACT implantation]. / Rektumperforation als Spätkomplikation einer ProACT-Implantation.

Operative approaches to solve severe urinary incontinence following radical prostatectomy include artificial urinary sphincters, bulking agents, urethral sling procedures, and the ProACT device (Uromedica Inc.). Previously reported complications of the ProACT procedure comprise ruptures and dislocations of balloons as well as erosions and perforations of the bladder or urethra. We report the case of a rectal perforation as a late complication of the ProACT procedure after prostatectomy and adjuvant radiotherapy. To the best of our knowledge this complication has not been described before.

Quantitative real-time RT-PCR of CD24 mRNA in the detection of prostate cancer.

BACKGROUND: Gene expression profiling has recently shown that the mRNA for CD24 is overexpressed in prostate carcinomas (Pca) compared to benign or normal prostate epithelial tissues. Immunohistochemical studies have reported the usefulness of anti-CD24 for detecting prostate cancer over the full range of prostate specimens encountered in surgical pathology, e.g. needle biopsies, transurethral resection of prostate chips, or prostatectomies. It is a small mucin-like cell surface protein and thus promises to become at least a standard adjunctive stain for atypical prostate biopsies. We tested the usefulness of real-time RT-PCR for specific and sensitive detection of CD24 transcripts as a supplementary measure for discriminating between malignant and benign lesions in prostatic tissues. METHODS: Total RNA was isolated from snap-frozen chips in 55 cases of benign prostatic hyperplasia (BPH) and from frozen sections in 59 prostatectomy cases. The latter contain at least 50% malignant epithelia. Relative quantification of CD24 transcripts was performed on the LightCycler instrument using hybridization probes for detection and porphobilinogen deaminase transcripts (PBGD) for normalization. RESULTS: Normalized CD24 transcript levels showed an average 2.69-fold increase in 59 Pca-cases (mean 0.21) when compared to 55 cases of BPH (mean 0.08). This difference was highly significant (p < 0.0001). The method has a moderate specificity (47.3%) but a high sensitivity (86.4%) if the cutoff is set at 0.0498. CD24 expression levels among Pca cases were not statistically associated with the tumor and lymph-node stage, the grading (WHO), the surgical margins, or the Gleason score. CONCLUSION: The present study demonstrates the feasibility of quantitative CD24 RNA transcript detection in prostatic tissues even without previous laser microdissection.