First Clinical Experience with Extracorporeally Induced Destruction of Kidney Stones by Shock Waves.

We performed extracorporeally induced destruction of kidney stones on 72 patients. No complications have resulted from the tissue exposure to high energy shock waves. Clearance studies before and after the shock wave treatment indicate no changes in renal function. The method was used successfully in all patients with stones in the renal pelvis. In none of these patients was an open operation required. Two patients with ureteral stones also were treated with shock waves but had to be operated upon because of insufficient destruction of the stone.

Stone/tissue differentiation for Holmium laser lithotripsy using autofluorescence: Clinical proof of concept study.

BACKGROUND AND OBJECTIVES: Holmium laser lithotripsy is the gold standard for intracorporeal fragmentation of urinary calculi. Usually, a visible beam is superimposed on the IR treatment laser as an aiming beam to guide the surgeon. In vitro tests showed that this aiming beam (532 nm, power <1 mW) excites strong fluorescence on human calculi. Tissue, in contrast, emitted much weaker fluorescence. If this is verified in vivo, the fluorescence signal induced by the aiming beam could be used to implement a feedback loop, preventing the Holmium laser being fired on tissue. MATERIALS AND METHODS: Fluorescence signals of 67 tissue and 68 stone spots were measured in a clinical proof of concept study with eight patients. For this, a modulated excitation/detection scheme (lock-in technique) was implemented. A frequency-doubled, diode-pumped solid-state laser module (532 nm, modulation frequency 66 Hz, average power 0.3 mW) was coupled via a dichroic mirror with the Holmium lithotripsy laser into the treatment fiber. The fluorescence signal entering the treatment fiber was detected via another dichroic mirror with a photodiode and a lock-in amplifier. RESULTS: In most instances (94%), the calculus of a patient gave a signal which was at least twice the maximum signal of ureteral tissue. CONCLUSION: The results of our proof of concept study indicate that measuring the fluorescence signal of a green aiming beam could be used to implement a feedback loop for Holmium laser lithotripsy. Preventing the laser being fired on tissue, this would increase the safety of the procedure. Lasers Surg. Med. 49:361-365, 2017. © 2016 Wiley Periodicals, Inc.

Stone/tissue differentiation during intracorporeal lithotripsy using diffuse white light reflectance spectroscopy: In vitro and clinical measurements.

BACKGROUND AND OBJECTIVE: Holmium laser lithotripsy is the 'gold standard' for intracorporeal fragmentation of stones. However, there is a risk of damaging and perforating the ureter wall when the laser is accidentally fired while the fiber is in contact with tissue. The aim of this study was to evaluate if white illumination light, diffusely reflected back into the treatment fiber and spectrally analyzed, can be used for differentiating between stone and tissue. STUDY DESIGN/MATERIALS AND METHODS: Firstly, in vitro reflectance spectra (Xenon light source, wavelength range λ = 350-850 nm) of 38 human kidney stones, porcine renal calix and ureter tissue were collected. Secondly, in an in vivo study with 8 patients, 72 ureter and 49 stone reflectance signals were recorded during endourological interventions. The spectra were analyzed to discriminate between stone and tissue by the absence or presence of minima due to hemoglobin absorption at λ1 = 542nm and λ3 = 576nm. RESULTS: In vitro, all stone and tissue signals could correctly be identified by calculating the ratio R = I (λ1 = 542 nm)/I (λ2 = 475 nm): Because of the hemoglobin absorption at λ1 , R is smaller for tissue than for calculi. In vivo, only 75% tissue spots could correctly be identified utilizing this method. Using the more sophisticated evaluation of looking for minima in the diffuse reflectance spectra at λ1 = 542 nm and λ3 = 576 nm, 62 out of 64 tissue spots were correctly identified (sensitivity 96.9%). This was also the case for 39 out of 43 stone spots. Taking into account the number of measured spectra, a tissue detection probability of 91% and a stone detection probability of 77% was achieved (significance level 5%). CONCLUSION: White light diffusely reflected off the treatment zone into the fiber can be used to strongly improve the safety of Holmium laser lithotripsy by implementing an automatic feedback control algorithm that averts mispositioning the fiber.

Could titanium oxide coating from a sol-gel process make stone baskets more resistant to laser radiation at 2.1 µm?

BACKGROUND: Stone baskets could be easily destroyed by Holmium:YAG-laser at an endourologic treatment, with respect to this, we try to improve the resistance by coating them with a titanium oxide layer. The layer was established by a sol-gel-process. MATERIALS AND METHODS: Six new baskets (Equadus, Opi Med, Ettlingen, Germany) were used: 1.8 Ch. with 4 wires (diameter 0.127 mm). Three baskets were coated with a layer of titanium oxide established by a sol-gel process at the BioCerEntwicklungs GmbH in Bayreuth (~100 nanometres thickness). The lithotripter was a Holmium:YAG laser (Auriga XL, Starmedtec, Starnberg, Germany). 10 uncoated and 10 coated wires were tested with 610 mJ (the minimal clinical setting) and 2 uncoated and 2 coated wires were tested with 110 mJ. The wires were locked in a special holding instrument under water and the laser incident angle was 90°. The endpoint was gross visible damage to the wire and loss of electric conduction. RESULTS: Only two coated wires resisted two pulses (one in the 610 mJ and one in the 110 mJ setting). All other wires were destroyed after one pulse. CONCLUSION: This was the first attempt at making stone baskets more resistant to a Holmium:YAG laser beam. Titanium oxide deposited by a sol-gel-process on a titanium-nickel alloy did not result in better resistance to laser injuries.

Laparoscopic adrenalectomy in urological centres - the experience of the German Laparoscopic Working Group.

OBJECTIVE: To evaluate the safety and feasibility of laparoscopic adrenalectomy (LA) performed in several German centres with different laparoscopic experience, as LA has become the gold-standard approach for benign surgical adrenal disorders; however, for solitary metastasis or primary adrenal cancer its precise role is uncertain. PATIENTS AND METHODS: The data of 363 patients who underwent a LA were prospectively collected in 23 centres. All centres were stratified into three groups according to their experience: group A (<10 LAs/year), group B (10-20 LAs/year) and group C (>20 LAs/year). In all, 15 centres used a transperitoneal approach, four a retroperitoneal approach and four both approaches. Demographic data, perioperative and postoperative variables, including operating time, surgical approach, tumour size, estimated blood loss, complications, hospital stay and histological tumour staging, were collected and analysed. RESULTS: The transperitoneal approach was used in 281 cases (77.4%) and the retroperitoneal approach was used in 82 patients (22.6%). In all, 263 of 363 lesions (72.5%) were benign and 100 (27.5%) were malignant. The mean (sd) operating time was 127.22 (55.56) min and 130.16 (49.88) min after transperitoneal and retroperitoneal LA, respectively. The mean complication rates for transperitoneal and retroperitoneal LA were 5% and 10.9%, respectively. CONCLUSION: LAs performed by urologists experienced in laparoscopy is safe for the removal of benign and malignant adrenal masses. LA for malignant adrenal tumours should be performed only in high-volume centres by a surgeon performing at least >10 LAs/year.

Comparison between a 1.92-µm fiber laser and a standard HF-dissection device for nephron-sparing kidney resection in a porcine in vivo study.

Nephron-sparing surgery was performed in a porcine model with a 1.92-µm fiber laser dissection device in comparison to a standard high-frequency dissection device. In nine pigs, general anesthesia and a median laparotomy were performed to expose both kidneys. On six kidneys (three HF and three laser) a partial renal parenchyma resection of the lower pole without opening of the renal pelvis was performed (group A). On 12 kidneys (four HF and eight laser), a hemi nephrectomy with opening of the renal pelvis was performed (group B). Total resection time including hemostasis of the remaining tissue was 501 ± 394 s in group "A-laser " vs. 176 ± 139 s in group "A-HF". For the group "B", the total resection time was 1174 ± 501 s (B laser) vs. 960 ± 407 s (B-HF). Blood loss was 28 ± 22 ml in group "A laser " vs. 15 ± 15 ml in group "A-HF". In group "B", the blood loss was 98 ± 73 ml (B laser) vs. 137 ± 118 ml (B-HF). No ischemic time for the kidneys was needed in group "A" for both dissection devices. In group "B", ischemia of the kidneys was performed three times during the eight laser procedures (420 ± 60 s) and only once at the four HF procedures (1,260 s). Healing process was observed over 4-6 weeks, survival rate was 100%, and no renal fistulas were found after the survival period. In conclusion, no significant differences were found between the compared dissection devices. However, the laser system with the flexible transmission fiber may have an advantage for a laparoscopic approach by steerable instruments.

In vivo studies on the availability and toxicity of antisense oligonucleotides in bladder cancer.

BACKGROUND: The urinary bladder is an ideal organ for topical treatment. A substantial number of bladder cancer patients are resistant to conventional intravesical therapy. In search of new agents, antisense oligonucleotides (AS-ON) may be interesting candidates. The availability and toxicity as well as the effectivity of AS-ON after intravesical instillation in different rodent models were examined. MATERIALS AND METHODS: Acute toxicity of AS-ON was tested by intravenous application (215-1,000 mg/kg body weight (bw)) in NMRI mice (n=30). The uptake and distribution of isotope-labelled AS-ON in bladder tissue was determined in Sprague Dawley rats (n=12) by radioactivity after intravesical application (2.5 mg/kg bw 3H-labelled AS-ON). Additionally, uptake and effectivity studies of AS-ON in tumors were performed in MB-49 bladder cancer-bearing C57/B16 mice (n=6) by immunohistochemistry and fluorescence microscopy. RESULTS: No systematic side-effects were noticed after intravenous application of physiological doses of AS-ON in NMRI mice. The mortality rate was 20% at the highest dose of 1,000 mg/kg bw. The highest AS-ON availability after intravesical application in rats was noticed in the bladder wall (12.3 microg/g), while the systemic concentration was low (1.1 microg/g). In fluorescence microscopy analysis, AS-ON were detected in the outer cells of the bladder wall and around vessels. AS-ON accumulated in the cytoplasm and in the nuclei. Immunohistochemical analysis demonstrated a reduction of the Ki-67 positivity after treatment with AS-ON (43%) compared to the untreated controls (58%). CONCLUSION: These preclinical experiments have shown that intravesical antisense oligonucleotides are safe and accumulate in the bladder and in bladder tumors, whereas systemic concentrations remain low. These data are the basis of a first clinical phase I study with intravesical instillation of Ki-67 antisense oligonucleotides.

[Physician health: What makes us ill? What keeps us healthy?] / Arztgesundheit: Was macht uns krank ­ Was hält uns gesund?

The medical profession is a particular health risk. Internal reasons for this are the common practice of self-diagnosis and self-therapy, presenteeism (work despite illness), and increased risks of addiction and suicide. External reasons include infectious diseases, violence against health professionals, the increasingly difficult working conditions - due to economisation, among other things - and the posttraumatic stress syndrome. Therefore, it is increasingly important to proactively take care of one`s own health. Science has shown that the underlying mechanisms leading to this problem are the high self-demand and lack of self-care observed in medical professionals. These starting points and available programs (cognitive-emotional reflection, stress management, relaxation techniques, conflict resolution techniques, mindfulness, priming, framing, meditation, embodiment, etc.) to strengthen resilience offer instruments to optimise the salutogenesis of health professionals. This review article presents interdependencies and concepts aiming to strengthen the resilience of health professionals.

Antisense-mediated inhibition of survivin, hTERT and VEGF in bladder cancer cells in vitro and in vivo.

Since cancer cells are characterised by multiple genetic alterations the single inhibition of one tumour- associated gene might not be sufficient as a therapeutic strategy. We examined the effects of a combined inhibition of survivin, human telomerase reverse transcriptase (hTERT) and vascular endothelial growth factor (VEGF) with antisense oligodeoxynucleotides (AS-ODNs) and small interfering RNAs (siRNAs) in EJ28 and 5637 bladder cancer (BCa) cells. Following verification of the uptake of intraperitoneally applied fluorescence-labelled AS-ODNs and siRNAs in subcutaneous BCa xenografts, the target-directed constructs were tested as single agents in SCID mice bearing subcutaneous EJ28. Simultaneous inhibition of two of the selected transcripts significantly enhanced cell viability reduction compared to the controls consisting of a target directed construct and an appropriate control construct without any homology to the human genome. The uptake of both antisense inhibitor types in the subcutaneous BCa was achieved even without a carrier. In vivo studies with 9 consecutive intraperitoneal injections with 20 mg/kg AS-ODNs or 4.6 mg/kg siRNAs revealed the biocompatibility of both antisense inhibitor types and showed anti-tumoural activity of the AS-ODNs used.

Prostate cancer vaccines: current status and future potential.

Standard systemic treatment of prostate cancer today is comprised of antihormonal and cytostatic agents. Vaccine therapy of prostate cancer is principally attractive because of the presence of tumor-associated antigens such as prostate-specific antigen (PSA), prostatic acid phosphatase (PAP), prostate-specific membrane antigen (PSMA), and others. Most prostate cancer vaccine trials have demonstrated some activation of the immune system, limited clinical success, and few adverse effects.One strategy to overcome the problem of limited clinical success of vaccine therapies in prostate cancer could be strict patient selection. The clinical course of patients with prostate cancer (even in those with PSA relapse following surgery or radiotherapy with curative intention, or those with metastatic disease) can vary significantly. In patients with organ-confined prostate cancer, the most promising immunotherapeutic approach would be an adjuvant therapy following surgery or radiotherapy. Patients with PSA relapse following surgery or radiotherapy could also benefit from immunotherapy because tumor burden is usually low. However, most patients in prostate cancer vaccine trials had metastatic hormone-refractory prostate cancer (HRPC). High tumor burden correlates with immune escape phenomena. Nevertheless, 2 years ago, it was demonstrated, for the first time, that a tumor vaccine can prolong survival compared with placebo in patients with HRPC. This was demonstrated with the vaccine sipuleucel-T (APC-8015; Provenge), a mixture of cells obtained from the patient's peripheral blood by leukapheresis followed by density centrifugation and exposition. The Biologics License Application for this vaccine was denied by the US FDA in mid 2007, however, because the trial had failed to reach the primary endpoint (prolongation of time to tumor progression). Nevertheless, clinical trials with sipuleucel-T are ongoing, and the approach still looks promising. Another interesting approach is a vaccine made from whole tumor cells: GVAX. This vaccine is presently being studied in phase III trials against, and in combination with, docetaxel. The results from these trials will become available in the near future. Besides the precise definition of the disease status of patients with prostate cancer, combinations of vaccine therapy with radiotherapy, chemotherapy, and/or hormonal therapy are approaches that look promising and deserve further investigation.

Drug evaluation: Therion's rV-PSA-TRICOM + rF-PSA-TRICOM prime-boost prostate cancer vaccine.

Therion Biologics Corp is developing PROSTVAC-VF-TRICOM, a prime-boost vaccine regimen that consists of a priming injection with a recombinant attenuated vaccinia virus expressing PSA and TRICOM (the company's proprietary triad of costimulatory molecules: ICAM-1, B7.1 and lymphocyte function-associated antigen-3), and a booster injection with a fowlpox virus expressing the same combination, for the potential treatment of prostate cancer. Phase II clinical trials are underway.

Bi-layer sandwich film for antibacterial catheters.

Background: Approximately one quarter of all nosocomial infections can be attributed to the urinary tract. The infections are supposed to be mainly caused by implantations of urethral catheters and stents. A new catheter design is introduced with the aim to lower the high number of nosocomial urethral infections. In order to avoid limitations to use, the design is first applied to conventional commercially available balloon catheters. Results: The main feature of the design is a sandwich layer on both sides of the catheter wall, which is composed of a fragmented base layer of silver capped by a thin film of poly(p-xylylene). This top layer is mainly designed to release a controlled amount of Ag+ ions, which is bactericidal, but not toxic to humans. Simultaneously, the lifetime is prolonged to at least one year. The base layer is electrolessly deposited applying Tollens' reagens, the cap layer is deposited by using chemical vapor deposition. Conclusion: The three main problems of this process, electroless deposition of a fragmented silver film on the surface of an electrically insulating organic polymer, irreproducible evaporation during heating of the precursor, and exponential decrease of the layer thickness along the capillary, have been solved trough the application of a simple electrochemical reaction and two standard principles of physics: Papin's pot and the principle of Le Chatelier.

Efficacy of silver/hydrophilic poly(p-xylylene) on preventing bacterial growth and biofilm formation in urinary catheters.

Catheter associated urinary tract infections (CAUTI), caused by several strains of bacteria, are a common complication for catheterized patients. This may eventually lead to a blockage of the catheter due to the formation of a crystalline or amorphous biofilm. Inhibiting bacteria should result in a longer application time free of complaints. This issue has been investigated using an innovative type of silver-coated catheter with a semipermeable cap layer to prevent CAUTI. In this work, two different types of silver catheters were investigated, both of which were capped with poly(p-xylylene) (PPX-N) and exhibited different surface properties that completely changed their wetting conduct with water. The contact angle of conventionally deposited PPX-N is approximately 80°. After O2 plasma treatment, the contact angle drops to approximately 30°. These two systems, Ag/PPX-N and Ag/PPX-N-O2, were tested in synthetic urine at a body temperature of 37 °C. First, the optical density and the inhibition zones of both bacteria strains (Escherichia coli and Staphylococcus cohnii) were examined to confirm the antibacterial effect of these silver-coated catheters. Afterward, the efficacy of silver catheters with different treatments of biofilm formed by E. coli and S. cohnii were tested with crystal violet staining assays. To estimate the life cycles of silver/PPX-catheters, the eluted amount of silver was assessed at several time intervals by anodic stripping voltammetry. The silver catheter with hydrophilic PPX-N coating limited bacterial growth in synthetic urine and prevented biofilm formation. The authors attribute the enhanced bacteriostatic effect to increased silver ion release detected under these conditions. With this extensive preparatory analytic work, the authors studied the ability of the two different cap layers (without silver), PPX-N and oxygen plasma treated PPX-N, to control the growth of a crystalline biofilm by measuring the concentrations of the Ca2+ and Mg2+ ions after exposure of the catheters to saturated urine for 24 h. The higher concentrations of Ca2+ and Mg2+ in the precipitates on the PPX-N catheters indicates that the hydrophilic PPX-N coating is superior to the simple PPX-N coating, with regard to the formation of a crystalline biofilm. Moreover, hydrophilic PPX-N as a cap layer may promote wettability and increase silver ion release rate and thus reduce the adhesion of suspended crystals to the catheter. Reduced bacterial growth and reduced adhesion may help to prevent CAUTI.

Results of multiparametric transrectal ultrasound-based focal high-dose-rate dose escalation combined with supplementary external beam irradiation in intermediate- and high-risk localized prostate cancer patients.

PURPOSE: Clinical results of a biologic information-based focused dose escalation combined with dose de-escalation for the whole organ in external beam radiotherapy + high-dose-rate brachytherapy (HDR-BT) boost application for localized prostate cancer in a consecutively treated patient cohort. METHODS AND MATERIALS: One hundred thirty patients were treated with external beam radiotherapy (50 Gy) complementary to two multiparametric transrectal ultrasound-guided 15 Gy HDR-BT fractions. Real-time multiparametric transrectal ultrasound-based biologic planning for high-dose-rate boost dose planning used the summation of gray scale and Doppler sonography imaging + biopsy information. Target subvolumes received HDR-BT dose escalation up to 60 Gy/fraction. Dose-volume histogram parameters, organ at risks doses, and toxicity results were investigated. RESULTS: The median followup was 4.3 years, the median age was 68.62 years, and the mean initial prostate-specific antigen was 18.69 ng/mL. Low-, intermediate-, and high-risk constituted 69%, 21%, and 10% of the patients, respectively. The mean peripheral dose was 3.9 Gy per fraction. Prostate-specific antigen nadir was in 93% of the patients ≤1 ng/mL. Quality parameters were as follows: D90: 6.58 Gy, V100: 30.36%, V150: 9.96%, V200: 3.16%, uD0.1: 7.34 Gy, uD2: 9.34 Gy, rD01: 10.56 Gy, and rD2: 8.32 Gy, respectively. We observed G1, G2, G3 urinary toxicity in 17/130, 11/130, and 2/130 patients, respectively. Rectal toxicity: G1 and G2 occurred in 19/130 and 2/130 patients with mean dose values G1: 8.2 Gy and G2: 8.76 Gy. Analysis of variance test resulted in no correlation between toxicities and any other investigated factors. CONCLUSIONS: Focused extreme dose escalation with low prostate mean peripheral dose results in excellent long-term outcome data and very high focal boost doses and is causing no enhancement in late treatment toxicity.

Drug evaluation: Degarelix--a potential new therapy for prostate cancer.

Ferring Research Ltd and licensee Astellas Pharma Inc are developing the gonadotropin-releasing hormone antagonist degarelix as a potential subcutaneous treatment for prostate cancer.

Silver ions eluted from partially protected silver nanoparticles.

The most prominent character of a new type of antibacterial urological catheters is the zebra-stripe pattern of a silver film, which is plated electroless on their interior wall and capped by a very thin semipermeable layer of parylene. This design effectively controls the release rate of Ag(+) ions in artificial urine, which has been measured as function of time with optical emission spectroscopy. By evaluating the minimum inhibitory concentration against certain strains of bacteria with solutions of AgNO3 of known concentration with the method of optical density and applying this analysis to the silver-eluting catheters, it was shown that this moderation prolongs the period of their application significantly. But to act as antibacterial agent in chlorine-containing solutions, as in urine, the presence of urea is required to avoid precipitation of AgCl and to meet or even exceed the minimum inhibitory concentration of Ag(+). The quality of the silver depot layer was further determined by the deposition rate and its morphology, which revealed that the film consisted of grains with a mean size of 150 nm.

Adjuvant autologous renal tumour cell vaccine and risk of tumour progression in patients with renal-cell carcinoma after radical nephrectomy: phase III, randomised controlled trial.

BACKGROUND: Organ-confined renal-cell carcinoma is associated with tumour progression in up to 50% of patients after radical nephrectomy. At present, no effective adjuvant treatment is established. We aimed to investigate the effect of an autologous renal tumour cell vaccine on risk of tumour progression in patients with stage pT2-3b pN0-3 M0 renal-cell carcinoma. METHODS: Between January, 1997, and September, 1998, 558 patients with a renal tumour scheduled for radical nephrectomy were enrolled at 55 institutions in Germany. Before surgery, all patients were centrally randomised to receive autologous renal tumour cell vaccine (six intradermal applications at 4-week intervals postoperatively; vaccine group) or no adjuvant treatment (control group). The primary endpoint of the trial was to reduce the risk of tumour progression, defined as progression or death. All patients were assessed after standardised diagnostic investigations at 6-month intervals for a minimum of 4.5 years. FINDINGS: By preoperative and postoperative inclusion criteria, 379 patients were assessable for the intention-to-treat analysis. At 5-year and 70-month follow-up, the hazard ratios for tumour progression were 1.58 (95% CI 1.05-2.37) and 1.59 (1.07-2.36), respectively, in favour of the vaccine group (p=0.0204, log-rank test). 5-year and 70-month progression-free survival rates were 77.4% and 72%, respectively, in the vaccine group and 67.8% and 59.3%, respectively, in the control group. The vaccine was well tolerated, with only 12 adverse events associated with the treatment. INTERPRETATION: Adjuvant treatment with autologous renal tumour cell vaccine in patients with renal-cell carcinoma after radical nephrectomy seems to be beneficial and can be considered in patients undergoing radical nephrectomy due to organ-confined renal-cell carcinoma of more than 2.5 cm in diameter.

Technology evaluation: Onyvax-P, Onyvax.

Onyvax is developing Onyvax-P, an allogeneic whole-cell vaccine, for the potential treatment of prostate cancer. The vaccine is currently undergoing phase II clinical trials.

Flavopiridol, an inhibitor of cyclin-dependent kinases, induces growth inhibition and apoptosis in bladder cancer cells in vitro and in vivo.

Flavopiridol is a semi-synthetic flavone analog of the alkaloid, rohitukine, a compound from an Indian tree, Dysoxylum binectariferum. It has been shown to inhibit cyclin-dependent kinases (CDKs), causing cell cycle arrest and growth inhibition. Flavopiridol is reported to have cytotoxic activity against a wide range of cancer cell lines and has demonstrated its efficacy in several clinical trials. Flavopiridol seems a well-suited potential new agent for the treatment of bladder cancer. We, therefore, evaluated whether flavopiridol inhibits growth and induces apoptosis in bladder cancer cells and additionally examined the toxicity and efficacy of this drug in vivo in a rat bladder cancer model. The in vitro experiments showed an IC20 of 50-100 nM in all cell lines tested. However, there was a difference in the response with regard to the grading of the tumor cells at higher doses. The IC50 was found to be 150-350 nM in the well-differentiated RT4 and RTI12 cell lines after treatment with flavopiridol, in comparison to a IC50 of 1000 nMfor the poorly-differentiated cell lines T24 and SUP. After exposure to flavopiridol, all tumor cell lines underwent significant apoptosis in comparison to untreated cells, beginning at a dose of 50 nM flavopiridol. At high concentrations (500 nM) of flavopiridol, 80-90% of all cells showed severe apoptotic alterations. The treatment of rat urinary bladder cancer with flavopiridol demonstrated the best efficacy with an intermittent treatment of 0.1 mg/kg, 3 times weekly over a total of 3 weeks, resulting in 7/12 animals tumor-free and a trend for the remaining tumors to have lower stage and grade. There seems to be a small advantage in intermittent versus daily application of flavopiridol. In summary, our results indicated that flavopiridol could be a useful therapeutic agent for bladder cancer, inhibiting tumor growth, malignant progression and inducing apoptosis.

Self-adjuvanted mRNA vaccination in advanced prostate cancer patients: a first-in-man phase I/IIa study.

BACKGROUND: CV9103 is a prostate-cancer vaccine containing self-adjuvanted mRNA (RNActive®) encoding the antigens PSA, PSCA, PSMA, and STEAP1. This phase I/IIa study evaluated safety and immunogenicity of CV9103 in patients with advanced castration-resistant prostate-cancer. METHODS: 44 Patients received up to 5 intra-dermal vaccinations. Three dose levels of total mRNA were tested in Phase I in cohorts of 3-6 patients to determine a recommended dose. In phase II, 32 additional patients were treated at the recommended dose. The primary endpoint was safety and tolerability, the secondary endpoint was induction of antigen specific immune responses monitored at baseline and at weeks 5, 9 and 17. RESULTS: The most frequent adverse events were grade 1/2 injection site erythema, injection site reactions, fatigue, pyrexia, chills and influenza-like illness. Possibly treatment related urinary retention occurred in 3 patients. The recommended dose was 1280 µg. A total of 26/33 evaluable patients treated at 1280 µg developed an immune response, directed against multiple antigens in 15 out of 33 patients. One patient showed a confirmed PSA response. In the subgroup of 36 metastatic patients, the Kaplan-Meier estimate of median overall survival was 31.4 months [95 % CI: 21.2; n.a]. CONCLUSIONS: The self-adjuvanted RNActive® vaccine CV9103 was well tolerated and immunogenic. The technology is a versatile, fast and cost-effective platform allowing for creation of vaccines. The follow-up vaccine CV9104 including the additional antigens prostatic acid phosphatase (PAP) and Muc1 is currently being tested in a randomized phase IIb trial to assess the clinical benefit induced by this new vaccination approach. TRIAL REGISTRATION: EU Clinical Trials Register: EudraCT number 2008-003967-37, registered 27 Jan 2009.