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.

[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 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.

[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 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.

[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.

[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.

[Significance of neoadjuvant therapy before radical prostatectomy]. / Stellenwert der neoadjuvanten Therapie vor radikaler Prostatektomie.

Neoadjuvant therapy before radical prostatectomy should increase survival in patients. This is necessary especially in patients with adverse prognostic factors for locally advanced disease, because in this stage radical prostatectomy as the only treatment results in a significantly reduced rate of progression-free survival. The aim of neoadjuvant therapy protocols is to increase local tumor control because of possible downstaging effects of the tumor and to improve systemic control because of elimination of circulating tumor cells and possible micrometastases. This review discusses the present and future aspects of neoadjuvant therapies in detail. The neoadjuvant hormonal therapy prior to radical prostatectomy results in a significant downstaging that does not translate into prolonged disease-free survival. This observation was made for short-term (3 months) and long-term (8 months) hormonal therapy. Therefore, neoadjuvant hormonal therapy has only a cosmetic effect on the pathological results and should not be advocated any more. Newer protocols have shown that neoadjuvant chemotherapy or hormone chemotherapy is feasible. The results obtained in non-randomized trials with small numbers of patients do not allow analyzing the efficacy of these protocols. Theoretically, neoadjuvant chemotherapy, especially a taxane-based protocol, which has shown efficacy in hormone-refractory disease, could improve disease outcome. Clinical trials are underway to prove this hypothesis. In the future, new therapeutic strategies could also be used in the neoadjuvant setting. It can only be speculated if antibody protocols or gene therapy will be used in this respect. In conclusion, there is no standard neoadjuvant protocol prior to radical prostatectomy. Whether chemotherapy will set a new standard for care has to be elucidated by the ongoing clinical trials.

[Pseudoaneurysm of a subsegmental renal artery after percutaneous nephrolitholapaxy]. / Pseudoaneurysma eines Nierenarterienastes nach perkutaner Nephrolitholapaxie.

At postoperative day 10 of a percutaneous nephrolitholapaxy for a large stone in the right renal pelvis of a 78-year-old female patient, persistent gross hematuria, requiring blood transfusions, occurred. Selective renovasography showed a pseudoaneurysm of the branch of a segmental artery close to the lower renal calices. Subsequent selective placement of two "steel-coils" in the feeding artery through a coaxial catheter system resulted in complete and stable occlusion without loss of renal parenchyma. Thereafter, bleeding immediately stopped and the patient suffered no further complications. This tool of interventional radiology should be kept in mind before open surgery is considered.