Th.M. De Reijke

Embryonic stem cell-like cells derived from adult human testis

BACKGROUND Given the significant drawbacks of using human embryonic stem (hES) cells for regenerative medicine, the search for alternative sources of multipotent cells is ongoing. Studies in mice have shown that multipotent ES-like cells can be derived from neonatal and adult testis. Here we report the derivation of ES-like cells from adult human testis. METHODS Testis material was donated for research by four men undergoing bilateral castration as part of prostate cancer treatment. Testicular cells were cultured using StemPro medium. Colonies that appeared sharp edged and compact were collected and subcultured under hES-specific conditions. Molecular characterization of these colonies was performed using RT-PCR and immunohistochemistry. (Epi)genetic stability was tested using bisulphite sequencing and karyotype analysis. Directed differentiation protocols in vitro were performed to investigate the potency of these cells and the cells were injected into immunocompromised mice to investigate their tumorigenicity. RESULTS In testicular cell cultures from all four men, sharp-edged and compact colonies appeared between 3 and 8 weeks. Subcultured cells from these colonies showed alkaline phosphatase activity and expressed hES cell-specific genes (Pou5f1, Sox2, Cripto1, Dnmt3b), proteins and carbohydrate antigens (POU5F1, NANOG, SOX2 and TRA-1-60, TRA-1-81, SSEA4). These ES-like cells were able to differentiate in vitro into derivatives of all three germ layers including neural, epithelial, osteogenic, myogenic, adipocyte and pancreatic lineages. The pancreatic beta cells were able to produce insulin in response to glucose and osteogenic-differentiated cells showed deposition of phosphate and calcium, demonstrating their functional capacity. Although we observed small areas with differentiated cell types of human origin, we never observed extensive teratomas upon injection of testis-derived ES-like cells into immunocompromised mice. CONCLUSIONS Multipotent cells can be established from adult human testis. Their easy accessibility and ethical acceptability as well as their non-tumorigenic and autogenic nature make these cells an attractive alternative to human ES cells for future stem cell therapies.

Prognostic Factors and Percutaneous Nephrolithotomy Morbidity: A Multivariate Analysis of a Contemporary Series Using the Clavien Classification

PURPOSE We stratified factors affecting treatment morbidity, compared the outcomes of percutaneous nephrolithotomy procedures from a single department and provided evidence of treatment benefits when percutaneous nephrolithotomy is performed in an expert setting. MATERIALS AND METHODS Since the department became a dedicated endourological center in 2002 we grouped all percutaneous nephrolithotomy procedures into those performed before 2002 (group 1) and after 2002 (group 2). The modified Clavien classification was used to score morbidity. Independent variables with an influence on complications were studied including stone size, operating time, operative complications, dilation device, urine culture, group allocation and lithotripsy device. Contingency and logistic regression were used for univariate and multivariate analysis. RESULTS Of the 244 percutaneous nephrolithotomy procedures 68 comprised group 1 and 176 formed group 2. Statistical preoperative differences were patient age, the use of anticoagulants and positive urinary cultures. Group 1 had a complication rate of 56.8% and group 2 had a complication rate of 37.2%. There were significant differences between the groups (p = 0.007). Almost all complications were grade 1 to 2. On univariate analysis the influence variables were urine culture (OR 1.69), group allocation (OR 2.20), stone size (OR 2.28), dilation device (OR 4.8), lithotripsy device (OR 1.22), perioperative complications (OR 2.83) and surgical time (OR 1.87). On multivariate analysis the independent factors in the complicated outcome were stone size (OR 1.25), type of lithotripsy device (OR 1.35) and incidence of perioperative complications (OR 3.71). CONCLUSIONS The dedicated setting for percutaneous nephrolithotomy at our center resulted in decreased operative time, more uneventful procedures and decreased hospitalization time. The modified Clavien morbidity score is a reliable tool for more objective outcome comparisons after renal stone treatment.

New developments in castrate-resistant prostate cancer

Castrate‐resistant prostate cancer (CRPC) occurs when disease progresses in the presence of castrate levels of androgens and remains sensitive to further hormonal manipulation. For many years the treatment of CRPC was limited to the use of docetaxel for metastatic disease. However, this has recently changed with the approval of several new agents. Sipuleucel‐T, an immunotherapeutic vaccine, is now available in the US for patients with non‐metastatic CRPC and abiraterone, an oral enzyme inhibitor of androgen biosynthesis, as well as cabazitaxel, a cytotoxic chemotherapeutic, have been approved for the treatment of metastatic CRPC. Also, denosumab, a subcutaneous antibody, is now an option for the treatment of patients with CRPC with bone metastases, in addition to zoledronic acid, an intravenous bisphosphonate.

Strontium-89 chloride versus palliative local field radiotherapy in patients with hormonal escaped prostate cancer: A phase III study of the European organisation for research and treatment of cancer

OBJECTIVES To compare toxicity, subjective response rate, time to subjective progression and overall survival in patients with painful bone metastases of hormone-resistant prostate cancer (HRPC) treated with a single intravenous injection of 150MBq (4mCi) Strontium(89) Chloride (S) or palliative local field radiotherapy (R) with the usual radiotherapy regimen used at each centre. The costs of both treatments were also assessed. PATIENTS AND METHODS 101 patients were randomized to S and 102 to R. Time to event endpoints were compared with the Logrank test and Kaplan-Meier curves, in the intent-to-treat population (2-sided alpha=0.05). RESULTS Baseline characteristics of both groups were comparable. There was a borderline statistically significant difference in overall survival in favour of the local field radiotherapy (R: 11 months; S: 7.2 months; p=0.0457). There was no difference in progression-free survival or time to progression. Subjective response was seen in 34.7% in the S-arm and in 33.3% in the R-arm. A biochemical response was observed in 10% and 13% of the R- and S-groups, respectively. There was no difference in treatment toxicity between the two groups. CONCLUSION In symptomatic HRPC, pain treatment with local field radiotherapy is associated with a better overall survival compared to Strontium(89). The lower costs of local field radiotherapy also favour the use of this treatment in patients with HRPC. The reason for the apparent survival benefit of localised radiation treatment is not clear.

High-speed photographic evaluation of endoscopic lithotripsy devices

SummaryShock-wave generation and bubble formation occurring during endoscopic lithotripsy were studied using high-speed photography for various devices: a pulsed-dye laser, a Q-switched Nd: YAG lase and an electrohydraulic (EHL) apparatus. The three devices investigated generated gas bubbles that rapidly expanded and decayed. The maximal size of these bubbles was 5–8 mm for the pulsed-dye laser, 6 mm for the Q-switched Nd: YAG laser and up to 14 mm for the EHL device. The bubble size appeared to be governed mainly by the energy per pulse delivered by the lithotripsy device. The shock-wave pattern depended strongly on the type of device used; the 25-ns Q-switched Nd: YAG laser pulse generated a single pressure step, whereas the 1.5-μs pulsed-dye laser produced a train of shock fronts.

Optical Coherence Tomography in Bladder Cancer

The current diagnostic standard for grading and staging of non-muscle-invasive bladder cancer (NMIBC) is based on the histopathology obtained during transurethral resection of the bladder tumour (TURBT) or biopsies. Although considered reference standard, this technique has some limitations: First, no real-time intraoperative histological information is obtained on stage and grade. Real-time intraoperative histological information could be helpful when NMIBC is treated by electric coagulation or laser ablation. The urologist has to rely on his/her judgement of stage and grade since no tissue is harvested for histological confirmation. In addition, in many centres, patients with a history of low-grade, non-invasive NMIBC with small recurrent NMIBC are being followed up with regular cystoscopy, instead of having directly TURBT. Real-time histological confirmation during cystoscopy could be very helpful in these patients. Finally, carcinoma in situ (CIS) may appear as a red mucosal lesion, mimicking inflammation. Again, real-time histological information could help in discriminating between CIS and inflammation. CIS can easily be missed on cystoscopy due to its flat appearance. If urine cytology is suspicious for high-grade bladder cancer, random biopsies are taken from the bladder to confirm or exclude CIS. This method results in a high rate of unnecessary biopsies. Real-time histopathology can possibly guide the direction of biopsies and thus avoid taking unnecessary biopsies. These examples illustrate the need for a fast, effective minimally invasive tool for assessment of cancerous lesions in the bladder. Optical coherence tomography (OCT) is a technology that has the potential to provide real-time information on grade and stage of a bladder cancer lesion.

7 In vivo optisch coherente tomografie voor evaluatie van hogere urinewegurotheeltumoren – eerste resultaten van een pilotstudie

Tijdschrift voor Urologie mei 2013 nr. 3 Resultaten Zowel na behandeling met op TPCS2a gebaseerde fotodynamische therapie (PDT) alleen, als na behandeling met bleomycine of een van de controlechemotherapeutica alleen werd in alle 5 cellijnen een dosisafhankelijke inhibitie van celproliferatie gezien. Bij de combinatiebehandelingen werd enkel een significant (p < 0,001) synergistisch effect, dus een fotochemisch internalisatie-effect, geobserveerd voor PDT gecombineerd met bleomycine in de T24en de AY-27-cellijn.

Die Bedeutung der entzündlichen Reaktion bei der intravesikalen BCG-Therapie

Die intravesikale Bacillus Calmette-Guerin (BCG)-Therapie ist eine effiziente Behandlungsmethode bei Patienten mit Carcinoma in situ (Cis) bzw. oberflachlichem papillarem Blasentumor (Herr et al. 1986; Lamm et al 1991). Ziel der BCG-Therapie ist die Beseitigung von Tumorzellen mittels immunologischer Stimulation.