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.

[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.

Phosphatidylcholine-fatty acid membranes. I. Effects of protonation, salt concentration, temperature and chain-length on the colloidal and phase properties of mixed vesicles, bilayers and nonlamellar structures.

The phase and colloidal properties of phosphatidylcholine/fatty acid (PC/FA) mixed vesicles have been investigated by optical methods, acid-base titration, and theoretically as a function of temperature (5-80 degrees C), molar lipid ratio (0-1), lipid chain length (C14-C18), headgroup ionization (1.5 less than or equal to pH less than or equal to 10), vesicle concentration (0.05-32 mumol vesicle.dm-3, and ionic strength (0.005 less than or equal to J less than or equal to 0.25). Increasing the fatty acid concentration in PC bilayers causes the phase transition temperatures (at 4 less than or equal to pH less than or equal to 5) to rise until, for more than 2 FA molecules per PC molecule, the sample turbidity exhibits only two transitions corresponding to the chain-melting of the 1:2 stoichiometric complexes of PC/FA, and pure fatty acid. The former transition is into a nonlamellar phase and is accompanied by extremely rapid vesicle aggregation (with association rates on the order of Ca approximately 10(7) dm3.mol-1.s-1) and massive lipid precipitation. Fluid-phase vesicles with less than 2 FA per PC associate much more slowly (Ca approximately 10(3) dm3.mol-1.s-1), their aggregation being comparable to that of the ordered-phase liposomes. Under no conditions was the relation between the fatty acid concentration and the vesicle association rate for the fluid-phase vesicles linear. In contrast to the X-ray diffraction data, optical measurements reveal a 'pretransitional region' between the chain-melting temperature of the PC component and the temperature at which the gross transformation into a nonlamellar phase sets in. This is seen for all lipid mixtures investigated. On the relative temperature scale, lipids with different chain lengths behave qualitatively similarly; however, the effective association constants determined for samples of constant lipid concentration seem to decrease somewhat with the number of CH2 groups per chain. Fatty acid protonation, which yields electrically neutral bilayers, invariably increases the rate of vesicle association; we have measured, for example, Ca approximately 10(2) at pH approximately 7 and Ca approximately 10(7) dm3.mol-1.s-1 at pH approximately 4). Protonation of the phosphatidylcholine phosphate groups, which causes a net positive charge to accumulate on the lipid vesicles, initially increases (Ca approximately 10(8) dm3.mol-1.s-1) but ultimately decreases (Ca approximately 10(7) dm3.mol-1.s-1) the rate of association between PC/FA (1:2) mixed vesicles.(ABSTRACT TRUNCATED AT 400 WORDS)

[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.

[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.

[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.

Angiotensin II enhances insulin sensitivity in healthy volunteers under euglycemic conditions.

OBJECTIVE: It has been postulated that vasoconstrictors cause insulin resistance. This effect has been documented for epinephrine but not for angiotensin II (Ang II). The aim of this study was to investigate the effect of the latter on insulin sensitivity. DESIGN: In order to study the influence of subpressor doses of Ang II on insulin-mediated glucose uptake under euglycemic conditions, eight healthy volunteers were allocated in random order to sham infusion or infusion of Ang II (first 0.75 ng/kg per min and subsequently 1.5 ng/kg per min). In addition, in seven of the subjects Ang II was infused after 3 days of indomethacin pretreatment (150 mg/day). METHODS: Insulin-mediated glucose uptake (expressed as M value) was measured with the euglycemic clamp technique. Insulin levels were measured enzymatically, plasma renin activity, Ang II, aldosterone and C-peptide levels by radioimmunoassay, blood pressure by Dinamap and muscle blood flow by plethysmography. RESULTS: The M value after sham infusion was 7.81 +/- 1.52 mg/kg per min and after 1.5 ng/kg Ang II per min was 9.76 +/- 1.26 mg/kg per min (P < 0.001). Indomethacin pretreatment did not abolish the Ang II-induced rise in the M value. Mean arterial blood pressure during the euglycemic clamp was unchanged with sham infusion and the low dose of Ang II. It increased slightly with the higher dose of Ang II. Inferior limb muscle perfusion was higher after infusion of Ang II than after sham infusion; this effect was not obliterated by indomethacin pretreatment. CONCLUSIONS: Ang II increases insulin-mediated glucose uptake: that is, it enhances insulin sensitivity by mechanisms independent of prostaglandins. The observations are of potential relevance to the changes in insulin sensitivity in some forms of hypertension.

Contribution of polycyclic aromatic compounds to the carcinogenicity of sidestream smoke of cigarettes evaluated by implantation into the lungs of rats.

Particles and semivolatiles from sidestream smoke of cigarettes smoked on a smoking machine were collected by a filter combination consisting of a glass fibre filter and silanized polystyrene beads. The extract of the glass fibre filter was separated by a Sephadex LH-20 column chromatography into a fraction containing non-aromatic material plus polycyclic aromatic compounds (PAC) with 2 and 3 rings and a fraction consisting of PAC with 4 and more rings. To evaluate the carcinogenicity, both fractions as well as the semivolatiles were implanted into the lungs of Osborne-Mendel rats at a dose level of one cigarette per animal and compared with three dose levels of benzo[a]pyrene (BaP). The most pronounced carcinogenic effect of the sidestream smoke (100 ng BaP per cigarette) was caused by the fraction containing polycyclic aromatic hydrocarbons (PAH) with 4 and more rings (5 carcinomas of the lungs/35 rats). This fraction represents only 3.5% by weight of the total sidestream smoke condensate. By contrast, the semivolatile material did not provoke any tumors. Only a small contribution to the total carcinogenicity (1 carcinoma of the lungs/35 rats) was observed for the fraction containing non-aromatic material and 2- and 3-ring PAHs.

Urinary and faecal excretion of pyrene and hydroxypyrene by rats after oral, intraperitoneal, intratracheal or intrapulmonary application.

The urinary and faecal excretion of pyrene and 1-hydroxypyrene after oral (53.4%), intraperitoneal (3.1%), intratracheal (30-37%) and intrapulmonary application (0.003%) to rats has been determined by means of gas chromatography/mass spectrometry and the excretion rates were found to depend on the mode of application. With regard to the low urinary excretion rates, 1-hydroxypyrene seems not to be very suitable as a biological marker for PAH exposure to man.

Dermal oncogenicity study of 2-ethylhexyl acrylate by epicutaneous application in male C3H/HeJ mice.

A carcinogenicity bioassay of 2-ethylhexyl acrylate (2-EHA) was conducted by applying 25 microliters 86.5%, 21%, or 2.5% 2-EHA in acetone three times a week to the clipped dorsal skin of male C3H/HeJ mice (80 per group) over their lifetime. Another group was treated with a 43% 2-EHA solution for 24 weeks and thereafter observed for lifetime (stop-test). An untreated group and a group that received only the diluent acetone served as controls. Treatment-related changes in the skin indicative of irritation (scaling, scabbing, hyperkeratosis, hyperplasia) were found in all 2-EHA-treated groups. These lesions were reversible in the 43% group immediately after treatment was stopped, and in the 2.5% group after the 11th week of treatment. Only in the 86.5% and 21% test groups showing chronic irritative skin damage was there a high incidence of nepolastic skin lesions (papillomas, carcinomas, and melanomas) with no dose dependency. In contrast, no skin tumors were found in the control groups, in the group treated with 2.5% 2-EHA for lifetime or in the group treated with 43% 2-EHA for about 6 months and observed for lifetime.

Diagnostic value of additional systematic prostate biopsies in patients undergoing transurethral resection of the prostate.

OBJECTIVES: There are patients with obstructive voiding symptoms and suspicious screening parameters in whom prostate cancer (PC) cannot be excluded prior to transurethral resection of the prostate (TURP). The goal of our study was to assess the diagnostic value of systematic biopsies of the peripheral zone of the prostate performed during TURP. METHODS: Between 1990 and 1995, 132 patients (average age 69.5 +/- 7.4 years) with at least one suspicious screening parameter underwent a TURP and additional systematic prostate biopsies. Pathology reports were reviewed to verify whether PC was present in the TURP chips or in the biopsy cores. RESULTS: Histologic examination found benign prostatic hyperplasia in 52, prostatitis in 53, and PC in 27 patients. PC was detected only with TURP in 11 patients (40.8%). In 15 patients (55.6%), both TURP and prostate biopsies showed PC. There was only 1 patient (3.7%) with a positive biopsy in whom the examination of the resection chips did not detect PC. In patients with negative digital rectal examination and intermediate prostate-specific antigen levels, prostate-specific antigen density was not able to differentiate between benign and cancerous lesions. CONCLUSIONS: There seems to be a subgroup of patients where systematic and repeated prostate biopsies fail to detect PC prior to TURP. Although the increase in the detection rate through additional prostate biopsies of the peripheral zone is limited, we would recommend these biopsies in addition to TURP, especially if patients are eligible for further curative treatment options.

Rare metastases of signet ring cell carcinomas to the scrotum: report of two cases.

Metastases of signet ring cell carcinomas to the scrotum are rare. We present 2 patients with this kind of tumor. In 1 patient, the scrotal pathologic examination helped to detect an adenocarcinoma of the appendix with a signet ring cell component, with an extent that had not been apparent clinically. The other patient was seen at an advanced stage of signet ring cell carcinoma of the sigmoid colon following surgical therapy and palliative chemotherapy. The route of metastases seems to be via seeding along the testicular cord and via lymphatic dissemination.

Use of pulsed Nd:YAG laser in the ureter.

An Nd-YAG laser of 1064-nm wavelength, 8-nanosecond pulse duration, and single-pulse energy of 20 to 80 mJ can create shock waves with a peak pressure of 1000 bar in less than 4 nanoseconds. The safety of this laser has been demonstrated in cultured cells, whole blood, and porcine urothelium. Clinically, ureteral calculi have been removed from 24 of 27 patients.

Utility of 24-h ambulatory blood pressure measurement in a routine clinical setting of patients with chronic renal disease.

The objective of the study was to investigate the utility of 24 h ambulatory blood pressure measurement (ABPM) in patients with chronic renal disease (CRD). A retrospective audit was performed in an academic nephrology department. ABPMs obtained from 95 consecutive outpatients with nondiabetic renal disease were analysed for their power to stratify disease severity and to predict progression of renal insufficiency during 3 years follow-up. The average of 10 consecutive office blood pressure (BP) readings performed at baseline was used as a reference. Correlations and tabulated statistics were calculated. Baseline mean diurnal BP was 138/87 mmHg by ABPM and 146/92 mmHg by average office BP (56.8 vs 25.3% normotensives, P < 0.01). Mean serum creatinine was 154 +/- 109 micromol/l. Patients without a circadian BP pattern (61%) had significantly higher serum creatinine, urinary albumin excretion and subsequent progression rates of renal failure (P < 0.05). Systolic and pulse pressure ABP values correlated significantly with serum creatinine levels at baseline and 3 years thereafter, independent of daytime (r = 0.23-0.60; P < 0.05). Abnormal nocturnal systolic and mean ABP, and pulse pressure but not average day office BP were significantly associated with abnormal serum creatinine and the presence of coronary heart disease at baseline (relative risk 1.6-1.7, P < 0.05). In conclusion, physicians considered ABPM preferentially in patients with borderline-controlled BP. ABPM provided significant additional information relevant to the clinical decision making process compared with average office BP alone thereby justifying its use in specific situations.

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.

Total radical-trapping antioxidative capacity of plasma and whole blood chemiluminescence in patients with inflammatory and autoimmune rheumatic diseases.

The total radical-trapping antioxidative capacity (TRAC) of plasma was evaluated in samples from patients suffering from various inflammatory and autoimmune rheumatic diseases (n=104) and correlated with the phorbol ester-stimulated chemiluminescence (CL) of neutrophils and monocytes in unseparated blood. Plasma and blood samples from age- and sex-matched healthy volunteers (n=25) and from patients with non-rheumatic internal diseases (n=31) served as controls. A 2 to 10 fold increase in whole blood chemiluminescence was found in rheumatic patients, which paralleled 50-80% decreased levels of plasma TRAC-values. While significant correlations between CL and TRAC were determined for patients with inflammatory arthritic diseases no correlations were found with patients suffering from connective tissue diseases. Prednisolone treatment of individual patients increased plasma TRAC-values substantially and decreased elevated levels of phagocytic CL generation to that of healthy controls. The main potential application of the assays described here is for the convenient assessment of disease activity and progression in individual patients with rheumatic diseases.

On the analysis of CO(2), H(2)- and CO, H(2)-mixtures by water-gas potentiometry with solid electrolyte cells.

The potentiometric analysis of CO(2), H(2) and CO, H(2)O-mixtures using oxide ion-conducting solid electrolytes requires the adjustment of the water-gas equilibrium without side reactions in the high-temperature galvanic cell. Conventional cell designs suitable for the analysis of H(2), H(2)O and CO, CO(2)-mixtures are not applicable due to the insufficient gas residence times in the cells and the insufficient catalytical activity of the platinum electrodes. Solid electrolyte cells have to be modified by integrating of suitable catalysts. Under optimized conditions of gas velocity and cell temperature both gas systems can be analyzed only by measuring the cell tension U (=-emf) and temperature in the favorized temperature range around 813 degrees C. Here systematical errors of the component ratio or the mole fraction were smaller than 6%. Several fundamental requirements for the application of catalysts in solid electrolyte cells for the analysis of reactive water-gas mixtures are pointed out.

Electrocautery: principles and practice.

We explain the basic physics of transurethral resection of the prostate (TURP). Modifications of electrodes or high-frequency units that should improve TURP are discussed. Finally, we introduce Coagulating Intermittent Cutting (CIC), which is a TURP using a modified high-frequency generator. Results of in vitro experiments and the first data from a multicenter trial are demonstrated.