Application of an asymmetric helical tube reactor for fast identification of gene transcripts of pathogenic viruses by micro flow-through PCR.

We have established a fast PCR-based micro flow-through process consisting of a helical constructed tube reactor. By this approach we can detect transcripts of measles and human papilloma virus (HPV) by continuous flow allowing for reverse transcription (RT) and amplification of cDNA. The micro reaction system consisted of two columnar reactors for thermostating the different reaction zones of the RT process and the amplification. The PCR reactor was built by asymmetric heating sections thus realizing different residence times and optimal conditions for denaturation, annealing and elongation. The system concept is based on low electrical power consumption (50-120 W) and is suited for portable diagnostic applications. The samples were applied in form of micro fluidic segments with single volumes between 65 and 130 nL injected into an inert carrier liquid inside a Teflon FEP tube with an inner diameter of 0.5 mm. Optimal amplification for template lengths of 292 bp (lambda-DNA), 127 bp (measles virus) and 95 bp (HPV) was achieved by maximal cycle times of 75 s.

Direct comparison of enzyme measurements from dried blood and leukocytes from male and female Fabry disease patients.

Anderson-Fabry disease is an X-linked disorder that is caused by deficiency of the lysosomal enzyme alpha-galactosidase A. Symptoms include chronic progressive painful small-fibre neuropathy, cornea verticillata, renal failure and heart disease. Interestingly, female heterozygous patients may also show severe symptoms. After clinical suspicion, usually the determination of alpha-galactosidase activity in leukocytes is requested first. Alternatively, an enzymatic assay using dried blood specimens has been described. Dried blood samples require less material and are substantially more stable (several months at room temperature) than whole-blood specimens. To validate the new method and to asses its usefulness for diagnosis of female patients, enzyme activities of alpha-galactosidase, beta-galactosidase and beta-glucuronidase from 78 known Fabry patients were compared (29 males, 47 females) between both materials. In summary, the determination of alpha-galactosidase activity using dried blood and leukocytes as well as the ratio of alpha-galactosidase to beta-glucuronidase in dried blood can improve the diagnostic specificity in cases of female patients who are difficult to identify when only leukocyte enzyme activities are considered.

Allogeneic retrovirally transduced, IL-2- and IFN-gamma-secreting cancer cell vaccine in patients with hormone refractory prostate cancer--a phase I clinical trial.

BACKGROUND: The purpose of this vaccine study was to determine the safety and feasibility of vaccination with an allogeneic prostate carcinoma cell line, LNCaP, expressing recombinant interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) and to evaluate the efficacy of inducing tumor-specific immune responses in HLA-A2-matched patients with hormone refractory prostate cancer (HRPC). METHODS: In a dose-escalating phase I study, HLA-A2-matched HRPC patients received four vaccinations of irradiated allogeneic LNCaP cells retrovirally transduced to secrete IL-2 and IFN-gamma at study day 1, 15, 29 and 92 and subsequently every 91 days unless tumor progression was evident. RESULTS: Three patients receiving the first dose level (7.5 million cells) showed no evidence of dose-limiting toxicity or vaccine-related adverse events including autoimmunity. One of three patients receiving the second dose level (15 million cells) developed a transient self-limiting grade 3 local injection site reaction (ulceration) after the eighth vaccination. Vaccine-induced immune responses against a broad array of prostate tumor associated antigens were detected in all six patients. Two of the three patients receiving the higher dose showed a decline in serum prostate-specific antigen (PSA) values of more than 50%, with one patient remaining on protocol for 3 years. CONCLUSIONS: Immunisation with the allogeneic LNCaP/IL-2/IFN-gamma vaccine is safe and feasible without any dose-limiting toxicity or autoimmunity. A 50% PSA decline was achieved in two of the six patients. This encouraging data provides the scientific rationale for further investigation of the vaccine in a phase II trial.

[Abdominal colic caused by mature cystic teratoma of the ovary]. / Kolikartige Unterbauchbeschwerden durch reifes Ovarialteratom.

Colicky lower abdominal pain can have several causes. Besides the typical urological reasons, differential diagnosis should include surgical, internal, and gynecological causes. In particular, in cases of ambiguous calcifications on plain abdominal films - as shown in the presented case - a mature teratoma of the ovary should be taken into consideration.

Age, comorbidity and hypertensive co-medication do not affect cardiovascular tolerability of 10 mg alfuzosin once daily.

PURPOSE: We assessed in real-life practice the impact of age, cardiovascular comorbidity and co-medication on the tolerability and efficacy of 10 mg alfuzosin OD in men with lower urinary tract symptoms suggestive of benign prostatic obstruction. MATERIALS AND METHODS: A total of 6,523 men with a mean age of 64.7 years were enrolled in a 6-month open label study of 10 mg alfuzosin OD. They were stratified by age quartile (younger than 60, 60 to 64, 65 to 70 and older than 70 years), comorbidity (hypertension, ischemic heart disease and diabetes) and antihypertensive co-medication (diuretics, beta-blockers, angiotensin converting enzyme inhibitors, angiotensin II inhibitors and calcium channel antagonists). RESULTS: Alfuzosin was effective and well tolerated. Despite an increased prevalence of cardiovascular comorbidity and antihypertensive co-medication with age changes in blood pressure did not significantly differ among age groups. In controls, ie those with no cardiovascular comorbidity or co-medication, alfuzosin produced minimal decreases in sitting systolic (mean -2.6 to -2.8 mm Hg) and diastolic (mean -1.7 to -1.8 mm Hg) blood pressure. In men with cardiovascular comorbidity mean decreases in systolic (-3.5 to 5.8 mm Hg) and diastolic (-2.0 to -3.3 mm Hg) blood pressure remained marginal. Of the 6,523 exposed patients 19.3% withdrew from the study, mainly for adverse events (6.4%) or a lack of efficacy (5.3%), while 229 (3.5%) experienced serious adverse events and 1,558 (23.9%) reported at least 1 treatment emergent adverse event. The most commonly reported adverse event was dizziness/postural dizziness (4.8%). Hypotension/postural hypotension was uncommon (0.7%). Age, cardiovascular comorbidity and antihypertensive co-medication had no impact on the safety profile of 10 mg alfuzosin OD. CONCLUSIONS: Alfuzosin (10 mg) OD is effective and well tolerated, and it has marginal effects on blood pressure, including in elderly patients and those with hypertension, ischemic heart disease or diabetes and those receiving antihypertensive agents.

[Penile cancer--aftercare with results. How much is necessary?]. / Peniskarzinom - Nachsorge mit Konsequenz. Wie viel ist notwendig?

Penile cancer is a rare tumor entity but penile carcinoma is characterized by a high recurrence rate regarding local, lymphatic, and hematogenous recurrence. The critical period for tumor recurrence is in the first 5 years. Therapeutic options for tumor recurrence can be differentiated by the type of recurrence and the preceding therapy. The prognosis of local or small lymphatic recurrence-if detected early and diligently diagnosed-can be improved significantly by radical surgery. On the other hand, systemic therapy of advanced lymphatic recurrences and hematogenous metastases will influence disease progression only marginally. Based on these considerations, the follow-up of penile cancer should be risk adapted but close as suggested by our algorithm. With a reduced, but close follow-up we can offer our patients aftercare with the consequence of improved prognosis.

Nerve replacement strategies for cavernous nerves.

OBJECTIVE: This article reviews novel restorative therapies for cavernous nerves that may be used to replace resected cavernous nerves at the time of pelvic surgery. METHODS: A literature-based presentation (Medline search) on current nerve replacement strategies was conducted with emphasis on neurobiological factors contributing to the restoration of erectile function after cavernous nerve injuries. RESULTS: A promising alternative to autologous nerve grafts for extending the length of successful nerve regeneration are artificial nerve guides. The addition of neurotrophic factors, extracellular matrix components and Schwann cells has been shown to promote cavernous nerve regeneration. Neurotrophic factors can be incorporated in the scaffold or can be supplied by cells seeded into the stroma. The regenerative capacity of these cells can be further enhanced by genetic modification with neurotrophic factor encoding genes. CONCLUSIONS: Artificial nerve guides, especially biodegradable ones containing growth-promoting factors or cells, are a promising option for the repair of cavernous nerve lesions.

[Incidental carcinoma of the prostate: can we and should we recommend radical prostatectomy?]. / Inzidentelles Prostatakarzinom: Kann und soll die radikale Prostatektomie danach empfohlen werden?

INTRODUCTION: There is controversy regarding tumor control of incidental prostate cancer (PC). We evaluated in a large cohort if we can recommend radical prostatectomy after TURP. MATERIAL AND METHOD: In 52 (4.3%) from a total of 1207 patients undergoing radical prostatectomy the diagnosis had been made by TURP. In a retrospective analysis we evaluated morbidity, histopathological results, and tumor control of pT1a/b tumors. RESULTS: The number of incidentally detected PC decreased with time. In 5.8% in the TURP group and in 0.5% of the needle biopsy group, there was no residual tumor found (p<0.001). Morbidity was similar +/- TURP with the exception of operation time (206 vs 188 min) and catheter duration (19.3 vs 17.3 days). Postoperative continence was identical. There was no difference in tumor control for local recurrence-free survival and PSA-free survival with and without TURP. CONCLUSIONS: The rate of incidentally detected PC by TURP decreases over time, but in almost all cases we found clinically relevant cancer. TURP is not an adverse prognostic factor and morbidity is similar compared with patients who were diagnosed by needle biopsy. Our data confirm that we should recommend radical prostatectomy to patients who are candidates for further curative therapy.

[Nerve repair strategies for restoration of erectile function after radical pelvic surgery]. / Nerveninterponate zur Wiederherstellung der erektilen Funktion bei radikalen Beckenoperationen.

Iatrogenic cavernous nerve lesions occurring during radical pelvic surgery often lead to irreversible erectile dysfunction. The nerve defects after excision of the neurovascular bundles must be reconstructed by interposition grafting to supply a permissive scaffold for oriented axonal regrowth. The use of autologous nerve grafts for the repair of human cavernous nerves during radical prostatectomy has been controversial regarding the limited success achieved with bilateral nerve grafting. Artificial nerve guides consisting of natural or synthetic materials have been successfully used for bridging peripheral nerve defects. The combination with Schwann cells, neurotrophic factors and extracellular matrix components has been shown to promote cavernous nerve regeneration.

[Transurethral resection for benign prostatic hyperplasia. current developments]. / Transurethrale Resektion bei benigner Prostatahyperplasie. Aktuelle Entwicklungen.

Transurethral resection of the prostate still represents the "gold standard" in surgical treatment of symptomatic benign prostatic enlargement (BPE). New demands were made on transurethral electrosurgery with the appearance of alternative, minimal invasive procedures. Bleeding complications had to be reduced. Different attempts were made to improve the conventional TURP. Band loops and roller bars were developed in order to gain a more effective coagulation during the cutting process by a larger tissue contact. Major advances could be achieved by modifications of the high-frequency generator. Improved procedures like the "Coagulating-Intermittent-Cutting" (CIC), the "Instant-Response-", as well as the "Dry-Cut-Technology" combine cutting and coagulating effects to realize lowered bleeding and an efficient resection process. The introduction of bipolar currency flow opens the possibility of conventional TUR-P with saline solutions and thus to the theoretical avoidance of the TUR-syndrome. By the neoadjuvant use of 5alpha-reductase-inhibitors before planned TUR-P, bleeding- and irrigation fluid absorption should be reduced by lowering perfusion and the size of the prostate. Regarding these innovative approaches which improve the surgical standard and minimize the risk of perioperative complications, transurethral electrosurgery remains the method of choice in instrumental therapy of symptomatic BPE. In the future efficacy and costs will be comparable to those of the transurethral laser-resection or laser-vaporisation.

Postoperative acute confusional state in typical urologic population: incidence, risk factors, and strategies for prevention.

OBJECTIVES: To determine the incidence of, and predictors for, the acute confusional state (ACS) in older patients after urologic surgery. ACS is among the most common complications after surgery in older patients. It is associated with increased postoperative morbidity, longer hospital stays, and greater mortality. Agitation caused by ACS might have deleterious consequences in a large proportion of older patients, especially after urologic surgery. Only a few studies, of highly selected urologic procedures, have been reported, and all showed an astonishingly low percentage of patients with this distressing condition. METHODS: We examined 100 consecutive, older patients (age 60 years or older), prospectively, before and after urologic surgery, to determine both the incidence of, and the predictors for, ACS. RESULTS: Only 7 of the 100 patients developed postoperative ACS. The risk factors identified were preoperative cognitive deficits, pre-existing depression, impaired vision, and the operative time. CONCLUSIONS: These results suggest that postoperative ACS is relatively rare after urologic surgical procedures; however, patients who are likely to develop ACS can be identified, prompting consideration for prophylactic antidelirium care.

[Tissue engineering of erectile nerves]. / Tissue Engineering erektiler Nerven.

Dissection of the cavernous nerves eliminates spontaneous erections and may lead to irreversible erectile dysfunction due to degeneration of cavernous tissue. Novel procedures to reconstruct penile innervation include cavernous nerve interposition grafting and neurotrophic treatments to revitalize penile neural input, evaluated thus far in various preclinical models of cavernous nerve injury. Schwann cells crucially contribute to successful axonal regeneration by mechanical and paracrine mechanisms in the injured nerve, and Schwann cells seeded into guidance channels have been successfully employed to support regeneration in animal models of cavernous nerve injury. Gene therapy, tissue engineering, and reconstructive techniques have been combined to deliver neurotrophic factors and recover erectile function.

[The influence of different wound drainage systems in radical retropubic prostatectomy--a prospective randomized study]. / Der Einfluss verschiedener Drainagesysteme bei der radikalen retropubischen Prostatektomie--eine prospektiv randomisierte Studie.

INTRODUCTION: Wound drainage after radical prostatectomy is used to reduce postoperative hematomas and lymphoceles and to drain any leakage at the vesicourethral anastomosis. We evaluated two different suction-drainage systems regarding their efficacy. MATERIAL AND METHOD: Fifty-seven patients with radical retropubic prostatectomy and pelvic lymphadenectomy were prospectively randomized. Two drains were placed bilaterally, using the Ulmer drain (16, F, Unoplast A/S, Maersk Medical) with suction. Patients with Ulmer drain were randomized to removal with and without prior shortening of the drain. The third system was the Blake silicon drain with J-VAC system (19F, Ethicon) with continuous suction. RESULTS: Of the 57 patients, 19 were treated by J-VAC drainage (J-VAC), 19 received the Ulmer drain with drain shortening (Ulmer-with) prior to removal and 19 received the Ulmer drain without any drain shortening (Ulmer-without). Total drainage volume (mean) was statistically significantly different (p < 0.001) with 760 cc for J-VAC, 309 cc for Ulmer-without and 234 cc for Ulmer-with. Ultrasonography demonstrated 11 lymphoceles, with 5 occurring with J-VAC, 3 with Ulmer-with and 3 with Ulmer-without. These differences did not reach statistical significance (p = 0.67). Intervention was needed in 3 of the 11 patients with lymphoceles (2 with J-VAC, 1 with Ulmer-without). The mean drainage time was 3.8 days for J-VAC, 2.9 days for Ulmer-with and 2.5 days for Ulmer-without, which was statistically significant different (p = 0.005). CONCLUSIONS: Perioperative wound drainage after radical prostatectomy with pelvic lymphadenectomy is useful to reduce perioperative complications. Different wound drainage systems demonstrate different efficacy regarding drainage volume, drainage duration and lymphocele formation. Our data are in favor of the Ulmer drain system with shortening of the drain prior to removal.

Improved urinary continence after radical retropubic prostatectomy with preparation of a long, partially intraprostatic portion of the membraneous urethra: an analysis of 1013 consecutive cases.

PURPOSE: To improve the rate of full continence in our patients, we performed, since June 1997, a careful preparation of the distally intraprostatic part of the membranous urethra to obtain a long urethral stump for the vesicourethral anastomosis. PATIENTS AND METHODS: In all, 610 patients without (group 1) and 403 patients with (group 2) a long intraprostatic stump of the urethra were asked by a self-administered questionnaire about their continence status. The rate of positive surgical margins were compared as a marker of local tumour control. RESULTS: Full continence (no pads) was achieved in 76.02% in group 1 and in 88.84%, of all patients in group 2. Stress incontinence (SIC) I degrees was found in 12.46% and 7.44% respectively, SIC II degrees was noted in 8.69 and 3.72% and complete incontinence was seen in 2.79% in group 1 and in two patients (0.5%) in group 2. Also the time to reach the final continence status was statistically and highly significantly (P<0.001) shortened. The rate of positive margins decreased in group 2, despite intraprostatic preparation. CONCLUSIONS: The preparation of a long, partially intraprostatic portion of the membranous urethra for vesicourethral anastomosis in radical retropubic prostatectomy leads to a statistically highly significant improvement of full continence and earlier continence in prostate cancer patients without compromising local tumour control.