Advanced vacuolation indicates propagation of various salmonid alphavirus type 2 isolates in Acholeplasma-infected BF-2 cells.

During previous routine inspections of bluegill fry (BF-2) and rainbow trout gonad (RTG-2) cells incubated with organ samples from asymptomatic Arctic char Salvelinus alpinus, brook trout Salvelinus fontinalis, and rainbow trout Oncorhynchus mykiss, a distinctive, reproducible cytopathic effect (CPE) appeared. The striking CPE, involving progressive vacuolation turning into slowly proceeding pyknotic degeneration, was originally attributed exclusively to enhanced growth of Acholeplasma sp. However, at a recent re-examination of re-infected BF-2 cells using electron microscopy (EM), conventional PCR, and quantitative PCR (qPCR), a virus was also detected. Two days post inoculation (dpi), EM revealed characteristic virions inside cytoplasmic vacuoles and next to bacteria outside the cells. The nucleotide sequences of the viral nsP3 gene fragment obtained from supernatants of infected cells were 100% identical and representative for salmonid alphavirus type 2 (SAV 2). The 16S RNA gene (16S rDNA) fragment sequences of the Mollicutes-specific PCR product obtained from SAV-infected as well as virus-free BF-2 control cells were identical with Acholeplasma laidlawii. In addition, qPCR results indicated enhanced propagation of virus and bacteria increasing with vacuolation between 5 and 8 dpi. Advanced vacuolation can be regarded as a CPE of both SAV and A. laidlawii, suggesting a viral impact on the bacterial infection that turns a latent intracellular stage into an apparent degenerative condition.

Comparison of adjustable male slings and artificial urinary sphincter in the treatment of male urinary incontinence: a retrospective analysis of patient selection and postoperative continence status.

PURPOSE: To analyze and compare preoperative patient characteristics and postoperative results in men with stress urinary incontinence (SUI) selected for an adjustable male sling system or an artificial urinary sphincter (AUS) in a large, contemporary, multi-institutional patient cohort. METHODS: 658 male patients who underwent implantation between 2010 and 2012 in 13 participating institutions were included in this study (n = 176 adjustable male sling; n = 482 AUS). Preoperative patient characteristics and postoperative outcomes were analyzed. For statistical analysis, the independent T test and Mann-Whitney U test were used. RESULTS: Patients undergoing adjustable male sling implantation were less likely to have a neurological disease (4.5% vs. 8.9%, p = 0.021), a history of urethral stricture (21.6% vs. 33.8%, p = 0.024) or a radiation therapy (22.7% vs. 29.9%, p = 0.020) compared to patients that underwent AUS implantation. Mean pad usage per day (6.87 vs. 5.82; p < 0.00) and the ratio of patients with a prior incontinence surgery were higher in patients selected for an AUS implantation (36.7% vs. 22.7%; p < 0.001). At maximum follow-up, patients that underwent an AUS implantation had a significantly lower mean pad usage during daytime (p < 0.001) and nighttime (p = 0.018). Furthermore, the patients' perception of their continence status was better with a subjective complete dry rate of 57.3% vs. 22.0% (p < 0.001). CONCLUSIONS: Patients selected for an AUS implantation showed a more complex prior history and pathogenesis of urinary incontinence as well as a more severe grade of SUI. Postoperative results reflect a better continence status after AUS implantation, favoring the AUS despite the more complicated patient cohort.

First confirmation of salmonid alphavirus infection in Arctic char Salvelinus alpinus and in Austria.

To date, sleeping disease (SD) caused by salmonid alphavirus 2 (SAV 2) has been reported in freshwater rainbow trout Oncorhynchus mykiss and Atlantic salmon Salmo salar. This study describes for the first time the occurrence of SD in farm-reared Arctic char Salvelinus alpinus and the occurrence of SAV in Austria. Clinical symptoms were indicative of the disease, and the diagnosis was confirmed by histopathology, infectivity in first passages of CHSE-214 cells and PCR. The phylogenetic analysis of the amplified SAV-nonstructural protein-3 (nsP3) fragment revealed the affiliation to the SAV 2 genotype.

[Tularemia in a jogger woman after the attack by a common buzzard (Buteo buteo): A "One Health" case report]. / Tularämie nach Angriff eines Mäusebussards auf eine Joggerin: Ein "One Health"-Fallbericht.

INTRODUCTION: A female jogger was attacked by a common buzzard (Buteo buteo) and was scratched lightly at the back of the head. One week later she was taken ill with high fever and headache which was later diagnosed as ulcero-glandular tularemia in regional lymph nodes, caused by Francisella tularensis. Recovery was only achieved after several weeks of systemic antibiotic treatment (Gentamicin/ Ciprofloxacine). Tularemia is a well known zoonotic disease, called "rabbit fever", mainly affecting rabbits and hares, but also small rodents. Human infection occurs often following tick bites or bloodsucking insects, or in hunters or slaughterers handling infected animals. Bites by mice have also been reported as a cause of tularemia. For the first time we report this case of tularemia as a result of an attack by a bird of prey. We assume that the bird acted as a vector just carrying the F. tularensis on its claws or beak, but we cannot exclude an infection of the bird itself. Several other joggers had also been attacked by a common buzzard in the same area shortly after the above described event and one of these also became infected with F. tularensis.

[Urological aspects in patients with liver cirrhosis]. / Urologische Aspekte bei Patienten mit Leberzirrhose.

BACKGROUND: Patients with liver cirrhosis (LC) are multimorbid and have urological diseases that occur more often than in the general population. The short life expectancy and poor general health complicate diagnosis and treatment. Due to the rising prevalence of LC, the number of patients from this group will increasingly present to urological practices. TOPICS: There are only a few studies about urological diseases in LC which describe the diagnostic and therapeutic challenges in this patient group. LC patients have altered baseline PSA values, which must be considered in PSA monitoring as part of preventative prostate care. In addition, difficulties arise in diagnosis and treatment of bladder dysfunction, genitourinary infections/sepsis, hypogonadism, erectile dysfunction, and urological malignancies. The implementation of invasive/tumor surgical interventions depends on LC severity. The safest surgical approach possible should be used. CONCLUSION: The worldwide prevalence of LC continues to rise but urological diagnosis and therapy in affected patients is often complicated due to their poor performance status.

Butanol production from concentrated lactose/whey permeate: use of pervaporation membrane to recover and concentrate product.

In these studies, butanol (acetone butanol ethanol or ABE) was produced from concentrated lactose/whey permeate containing 211 g L(-1) lactose. Fermentation of such a highly concentrated lactose solution was possible due to simultaneous product removal using a pervaporation membrane. In this system, a productivity of 0.43 g L(-1) h(-1) was obtained which is 307 % of that achieved in a non-product removal batch reactor (0.14 g L(-1) h(-1)) where approximately 60 g L(-1) whey permeate lactose was fermented. The productivity obtained in this system is much higher than that achieved in other product removal systems (perstraction 0.21 g L(-1) h(-1) and gas stripping 0.32 g L(-1) h(-1)). This membrane was also used to concentrate butanol from approximately 2.50 g L(-1) in the reactor to 755 g L(-1). Using this membrane, ABE selectivities and fluxes of 24.4-44.3 and 0.57-4.05 g m(-2) h(-1) were obtained, respectively. Pervaporation restricts removal of water from the reaction mixture thus requiring significantly less energy for product recovery when compared to gas stripping.

The inter-individual variability outperforms the intra-individual variability of differentially expressed proteins prior and post irradiation in lymphoblastoid cell lines.

CONTEXT: Radio-sensitivity in normal tissue is characterized by heterogeneity throughout the population and the absence of pre-diagnostic biomarkers. OBJECTIVE: We conducted a proteomic approach to search for radiation characteristic protein regulation. MATERIALS AND METHODS: Cell lines were 10 Gy irradiated and analysed by 2D-DIGE after 24 h. RESULTS were analysed intra- and inter-individually. The principal component analysis and hierarchical clustering was applied to all datasets. RESULTS: Differences in intra-individual spot abundance prior and post irradiation exactly show the separation of sample classes in two groups: sham-irradiated and irradiated. The inter-individual datasets clustered according to the cell line. The intra-individual differences on protein level after gamma-irradiation are very low, compared with the inter-individual differences among cell lines derived from the same tissue. CONCLUSION: The application of 2-D DIGE may offer a realistic chance for a better molecular characterization of radio-sensitivity and for the discovery of candidate biomarkers.

[Urological comorbidities in patients with rheumatoid arthritis : literature review]. / Urologische Begleiterkrankungen bei Patienten mit rheumatoider Arthritis : Literaturübersicht.

Patients with rheumatoid arthritis (RA) have an increased risk of urolithiasis which is further negatively impacted by a reduced bone density. Interstitial cystitis also tends to occur more often in patients with rheumatic diseases. The high incidence of bacterial urogenital infections is influenced by the use of immunomodulating drugs. Many RA patients have to undergo numerous tests until a diagnosis is reached and are then treated as outpatients on a tightly controlled schedule. Despite a closely controlled rheumatological follow-up, urological screening and determination of a baseline prostate-specific antigen (PSA) value (in men over 45 years old) should not be neglected. In patients with an increased risk of renal and bladder neoplasms or when such a diagnosis is known, the benefit of long-term use of high doses of non-steroidal anti-inflammatory drugs (NSAID, aspirin type) should be carefully weighed up with a risk profile and after specialist urological assessment. Patients who suffer from sexual dysfunction due to physical limitations and prolonged medical therapy should undergo urological and gynecological assessment to exclude contributing causes. The use of aphrodisiacs and erection-enhancing drugs (e.g. PDE5 inhibitors, local injection with prostaglandins and vacuum therapy) require prior approval by a medical specialist and also cardiovascular stability. Acute urinary retention is more common in chronic inflammatory musculoskeletal diseases.

Subdiffusion supports joining of correct ends during repair of DNA double-strand breaks.

The mobility of damaged chromatin regions in the nucleus may affect the probability of mis-repair. In this work, live-cell observation and distance tracking of GFP-tagged DNA damage response protein MDC1 was used to study the random-walk behaviour of chromatin domains containing radiation-induced DNA double-strand breaks (DSB). Our measurements indicate a subdiffusion-type random walk process with similar time dependence for isolated and clustered DSBs that were induced by 20 MeV proton or 43 MeV carbon ion micro-irradiation. As compared to normal diffusion, subdiffusion enhances the probability that both ends of a DSB meet, thus promoting high efficiency DNA repair. It also limits their probability of long-range movements and thus lowers the probability of mis-rejoining and chromosome aberrations.

[Fever and lymphadenopathy. Report of 4 cases of tularemia]. / Fieber und Lymphadenopathie. Bericht über 4 Tularämiefälle.

We report on 4 patients with different types of tularemia acquired in Switzerland or nearby countries. All patients presented with fever, moderate to highly elevated signs of inflammation, and local lymphadenopathy. Additionally, 3 patients did not respond to empirical antimicrobial therapy with aminopenicillins. A tick bite was identified as mode of transmission in 2 patients, while 1 patient showed a possible connection to a tick bite. The route of transmission for the fourth patient remained unknown. The diagnosis of tularemia was either based on positive serology, on a positive polymerase chain reaction (PCR) from the lymph node samples or on positive blood cultures. The treatment in adult patients was ciprofloxacin 500-750 mg twice daily orally for 3 weeks. The pediatric patient was treated with gentamicin 4 mg/kg i.v. once daily for 1 week and ciprofloxacin 15 mg/kg twice daily orally for another 2 weeks. All patients recovered completely. Due to the increasing incidence of tularemia in Switzerland, this infection should be considered in patients with fever and lymph node enlargement particularly after tick bite. We recommend treatment with ciprofloxacin orally for 14-12 days.

An intensive 5-year-long influenza vaccination campaign is effective among doctors but not nurses.

OBJECTIVE: To double the vaccination rates of hospital employees from 20 to 40% by specific interventions over a 5-year period (2003-2007). The secondary endpoint was to compare the effects of the avian influenza in 2005 (intervention period) and the H1N1 influenza pandemic in 2009 (follow-up period, 2008-2009) on vaccination rates. DESIGN, SETTING, AND PARTICIPANTS: Free vaccination and its intensive propagation from 2003-2007 in a 400-bed teaching hospital with 1,687 hospital employees. Annual vaccination rates were obtained from 2003 through 2009. MAIN OUTCOME MEASUREMENTS: Yearly vaccination rates for the intervention period from 2003-2007 and the observational follow-up period of 2008-2009. RESULTS: The overall rate for seasonal influenza vaccination changed non-significantly during the intervention period from 20% in 2003 to 27% in 2007. At the end of the follow-up period in 2009, the vaccination rate was 26%, which was not significantly higher compared with that in 2003. Physicians interestingly increased from 34% in 2003 to 62% in 2007 and to 66% in 2009 (p < 0.001), while nurses dropped non-significantly from an already low proportion of 18% in 2003 to 15% in 2007 and to 16% in 2009 for seasonal influenza vaccination. The difference between nurses and doctors in 2007 is highly significant (p < 0.001). In the year of the avian influenza threat (2005), a significant increase was observed (30 vs. 20%, p < 0.001). This observation was seen again in 2009 (influenza A/H1N1v pandemic), during which the H1N1 vaccine uptake was 33% (p < 0.001, compared to seasonal flu vaccine in 2003). CONCLUSIONS: Overall, the vaccination rates did not increase over the 7-year study period. Interventions were successful for physicians but not for nurses. The vaccine uptake was significantly higher during the threat of avian influenza and the influenza A/H1N1v pandemic.

Kidney biopsy in patients with diabetes mellitus.

BACKGROUND: The clinical consequences of the results obtained by kidney biopsy in patients with diabetes mellitus Type 1 or Type 2 have been controversial. Our study was conducted to assess clinical symptoms and histological diagnoses in patients with diabetes mellitus Type 1 and Type 2 undergoing kidney biopsy. DESIGN, SETTING AND PATIENTS: Observational study. The study included data from 567 consecutive renal biopsies of patients with diabetes mellitus Type 1 or 2 and chronic kidney disease (CKD) examined by standard histopathological procedures. The main outcome measures were incidence of diabetic nephropathy (DN) and glomerulonephritis (GN), predictors for the presence of both DN or GN. RESULTS: Approximately 70% of patients with diabetes mellitus Type 1 or 2 and evidence for CKD had DN. Glomerular diseases present in approximately 30% of patients with diabetes were predominantly immune complex GN and secondary focal glomerulosclerosis, followed by IgA-GN, which was associated with microhematuria (p = 0.01) and hypertension (p = 0.04). Only a minority had membranous GN, which was associated with nephrotic syndrome (p = 0.004). Progressive CKD predicted the presence of GN in diabetes mellitus Type 2 (r = -0.98; p = 0.02). CONCLUSION: GN is not uncommon in patients with diabetes and evidence for CKD. Kidney biopsy should therefore be considered in patients with diabetes and progressive CKD.

The effectiveness of 20 mev protons at nanosecond pulse lengths in producing chromosome aberrations in human-hamster hybrid cells.

Laser accelerated radiotherapy is a potential cancer treatment with proton and carbon-ion beams that is currently under development. Ultra-fast high-energy laser pulses will accelerate ion beams that deliver their dose to a patient in a "pulsed mode" that is expected to differ from conventional irradiation by increasing the dose delivery rate to a tissue voxel by approximately 8 orders of magnitude. In two independently performed experiments at the ion microprobe SNAKE of the 14 MV Munich tandem accelerator, A(L) cells were exposed either to protons with 1-ns pulse durations or to protons applied over 150 ms in continuous irradiation mode. A slightly but consistently lower aberration yield was observed for the pulsed compared to the continuous mode of proton irradiation. This difference was not statistically significant when each aberration type was analyzed separately (P values between 0.61 and 0.85 in experiment I and P values between 0.32 and 0.64 in experiment II). However, excluding the total aberrations, which were not analyzed as independent radiation-induced effects, the mean ratio of the yields of dicentrics, centric rings and excess acentrics scored together showed (with 95% CI) a significant difference of 0.90 (0.81; 0.98) between the pulsed and the continuous irradiation modes. A similar tendency was also determined for the corresponding RBE values relative to 70 kV X rays. Since the different findings for the comparisons of individual chromosome aberration types and combined comparisons could be explained by different sample sizes with the consequence that the individual comparisons had less statistical power to identify a difference, it can be concluded that 20 MeV protons may be slightly less effective in the pulsed mode.

A case study of female genital tuberculosis in a Western European setting.

We have studied five women with genital tuberculosis (TB), which is an uncommon disease in Western European countries. However, young women that have emigrated from a country with high TB-prevalence and have complaints of unexplained infertility and nonspecific abdominal or pelvic symptoms should be checked for this disease. Because genital tuberculosis frequently causes infertility, early diagnosis and treatment are crucial, necessitating rapid microscopic, histological, and microbiological (culture) testing in suspected cases.

The WST survival assay: an easy and reliable method to screen radiation-sensitive individuals.

An easy, fast and reliable method was developed to screen hundreds of Epstein-Barr virus-transformed cell lines (lymphoblastoid cell lines, LCLs) for radiation sensitivity that were generated from lymphocytes isolated from young lung cancer patients. The WST-1 test explores the metabolic activity of the mitochondria as an indicator for the vital status of cells. Cell proliferation as well as indirect cell death can be quantified by this method on a large scale in microtiter plates. Cell survival was measured at 24- and 48-h post-irradiation with 10 Gy ((137)Cs source) by the WST-1 assay and Trypan blue staining. To set up the experimental screening conditions and to establish a positive and a negative control, an ATM-mutated cell line from a radiation-sensitive ATM patient and an ATM proficient cell line from a healthy brother were compared. An optimal differentiation between the two cell lines was demonstrated for 10 Gy and 24- and 48-h cell growth after irradiation. Upon screening 120 LCLs of young lung cancer patients under these conditions, 5 of them were found to be radiation sensitive to a high degree of statistical significance. The results have been confirmed by a second laboratory by means of Trypan blue testing. The WST-1 test represents an efficient and reliable method by means of screening for radiation-sensitive cell lines.

Heterogeneity of gene expression in stromal fibroblasts of human breast carcinomas and normal breast.

Breast carcinoma invasion is associated with prominent alterations in stromal fibroblasts. Carcinoma-associated fibroblasts (CAF) support and promote tumorigenesis, whereas normal mammary fibroblasts (NF) are thought to suppress tumor progression. Little is known about the difference in gene expression between CAF and NF or the patient-to-patient variability in gene expression. Paired CAF and NF were isolated from six primary human breast carcinoma specimens. RNA was extracted from low-passage cultures of CAF and NF and analyzed with Affymetrix Human Genome U133 Plus 2.0 arrays. The array data were examined with an empirical Bayes model and filtered according to the posterior probability of equivalent expression and fold difference in expression. Twenty-one genes (27 probe sets) were up-regulated in CAF, as compared with NF. Known functions of these genes relate to paracrine or intracellular signaling, transcriptional regulation, extracellular matrix and cell adhesion/migration. Ten genes (14 probe sets) were down-regulated in CAF, including the pluripotency transcription factor KLF4. Quantitative RT-PCR analysis of 10 genes validated the array results. Immunohistochemical staining for three gene products confirmed stromal expression in terms of location and relative quantity. Surprisingly, the variability of gene expression was slightly higher in NF than in CAF, suggesting inter-individual heterogeneity of normal stroma.

Postprandial blood glucose level in maintenance hemodialysis patients predicts post-transplant-diabetes-mellitus.

Post-transplant-diabetes-mellitus (PTDM) is a frequent complication after kidney transplantation. One-hundred-and-seven patients with kidney transplantation were screened for the occurrence of PTDM. Of these, full data sets from 49 subjects were available with documented glucose concentrations during maintenance hemodialysis (MHD) and regular clinical follow-up of 7-34 months. For assessment of glucose metabolism the response to a standard meal during MHD was used in normoglycemic patients based on fasting blood glucose. Abnormal postprandial blood glucose concentration was defined as >140 mg/dl 2 h after food intake.Twelve end stage renal disease patients had abnormal postprandial blood glucose on MHD. All 12 subjects but also four MHD patients with normal postprandial and fasting blood glucose values developed PTDM. Multivariate Cox-regression analysis revealed that abnormal postprandial blood glucose is a strong predictor for PTDM (Hazard ratio: 42.3 (IQR: 7.9-227.2); p<0.001). Fasting blood glucose (94 vs. 100 mg/dl) was not different between MHD patients who did (n=16) or did not (n=33) develop PTDM.This study suggests that measurement of postprandial blood glucose during MHD identifies patients who develop PTDM after kidney transplantation. It should be used for screening of patients at risk.

No evidence for a different RBE between pulsed and continuous 20 MeV protons.

To obtain greater insight into the future potential of tumor radiotherapy using proton beams generated from high-intensity lasers, it is important to characterize the ionization quality of the new beams by measuring the relative biological effectiveness (RBE) under conditions where the full dose at one irradiation site will be deposited by a few proton pulses less than 1 ns in duration. HeLa cells attached to a Mylar foil were irradiated with 70 kV X rays to obtain a reference dose-response curve or with 3 Gy of 20 MeV protons at the Munich tandem accelerator (Garching), either using a continuous mode where a cell sample was irradiated within a 100-ms time span or using a pulsed mode where radiation was given in a single proton pulse of about 1 ns. After irradiation cytochalasin B was added; 24 h later cells were fixed and stained with acridine orange and micronuclei were counted. The X-ray dose-response curve for the production of micronuclei in HeLa cells followed a linear-quadratic model. The corresponding RBE values for 20 MeV protons in pulsed and continuous irradiation modes were 1.07 +/- 0.08 and 1.06 +/- 0.10 in the first proton experiment and 1.09 +/- 0.08 and 1.05 +/- 0.11 in the second, respectively. There was no evidence for a difference in the RBE for pulsed and continuous irradiation of HeLa cells with 20 MeV protons.

Determination of glucose and ethanol in bioethanol production by near infrared spectroscopy and chemometrics.

The concentrations of glucose and ethanol in substrates from bioethanol processes have been modeled by near infrared (NIR) spectroscopy data. NIR spectra were acquired in the wavelength range of 1100-2300 nm by means of a transflectance probe for measurements in liquid samples. For building of regression models a genetic algorithm has been applied for variable selection, and partial least-squares (PLS) regression for creation of linear models. A realistic estimation of the prediction performance of the models was obtained by a repeated double cross-validation (rdCV). Reduced data sets with only 15 variables showed improved prediction qualities, in comparison with models containing 235 variables, particularly for the determination of the ethanol concentration in distillation residues (stillages). The squared correlation coefficient, R(2), between the concentrations obtained by HPLC analysis and the concentrations derived from NIR data (using 15 selected wavelengths, test set samples) was 0.999 for ethanol in stillage, and 0.977 for glucose in mash. The standard deviation of prediction errors, SEP, obtained from test set samples was 0.6 g L(-1) for ethanol (2% of the mean ethanol concentration), and 2.0 g L(-1) for glucose (9.6% of the mean glucose concentration).