Robert Koch InstituteBerlin

Robert, Germany

Robert Koch InstituteBerlin

Robert, Germany
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Ivanusic D.,Robert Koch InstituteBerlin | Ivanusic D.,Freie University BerlinBerlin | Heinisch J.J.,University of Osnabrück | Eschricht M.,Robert Koch InstituteBerlin | And 2 more authors.
BioTechniques | Year: 2015

Yeast-based methods are still the workhorse for the detection of proteinprotein interactions (PPIs) in vivo. Yeast two-hybrid (Y2H) systems, however, are limited to screening for a specific group of molecules that interact in a particular cell compartment. For this reason, the splitubiquitin system (SUS) was developed to allow screening of cDNA libraries of full-length membrane proteins for protein-protein interactions in Saccharomyces cerevisiae. Here we demonstrate that a modification of the widely used membrane SUS involving the transmembrane (TM) domain of the yeast receptor Wsc1 increases the stringency of screening and improves the selectivity for proteins localized in the plasma membrane (PM). © 2015 Eaton Publishing Company. All rights reserved.

Anoh A.E.,Alassane Ouattara University | Akoua-Koffi C.,Alassane Ouattara University | Couacy-Hymann E.,Laboratoire Central Of Pathologie Animale | Pauly M.,Robert Koch InstituteBerlin | And 10 more authors.
Virology Journal | Year: 2015

Background: Cytomegaloviruses (CMVs) are herpesviruses that infect many mammalian species, including humans. Infection generally passes undetected, but the virus can cause serious disease in individuals with impaired immune function. Human CMV (HCMV) is circulating with high seroprevalence (60-100 %) on all continents. However, little information is available on HCMV genoprevalence and genetic diversity in subsaharan Africa, especially in rural areas of West Africa that are at high risk of human-to-human HCMV transmission. In addition, there is a potential for zoonotic spillover of pathogens through bushmeat hunting and handling in these areas as shown for various retroviruses. Although HCMV and nonhuman CMVs are regarded as species-specific, potential human infection with CMVs of non-human primate (NHP) origin, shown to circulate in the local NHP population, has not been studied. Findings: Analysis of 657 human oral swabs and fecal samples collected from 518 individuals living in 8 villages of Côte d'Ivoire with generic PCR for identification of human and NHP CMVs revealed shedding of HCMV in 2.5 % of the individuals. Determination of glycoprotein B sequences showed identity with strains Towne, AD169 and Toledo, respectively. NHP CMV sequences were not detected. Conclusions: HCMV is actively circulating in a proportion of the rural Côte d'Ivoire human population with circulating strains being closely related to those previously identified in non-African countries. The lack of NHP CMVs in human populations in an environment conducive to cross-species infection supports zoonotic transmission of CMVs to humans being at most a rare event. © 2015 Anoh et al.

Pilatz A.,Justus Liebig University | Hossain H.,Justus Liebig University | Kaiser R.,University of Cologne | Mankertz A.,Robert Koch InstituteBerlin | And 6 more authors.
European Urology | Year: 2015

Background Acute epididymitis is a common infectious disease of unknown etiology in about 30% of cases with guidelines based on studies published >15 yr ago. Objective To investigate the etiology of acute epididymitis using state-of-the-art methods and to provide rational data for antimicrobial therapy and clinical management. Design, setting, and participants Between 2007 and 2013, 237 patients (150 antimicrobially naive and 87 antibiotically pretreated) with acute epididymitis underwent comprehensive investigation comprising microbiologic cultures, polymerase chain reaction (PCR) for sexually transmitted infections (STIs), 16S ribosomal DNA (rDNA) analysis, and PCR detection of 23 viruses. Clinical management followed international guidelines. Outcome measures and statistical analysis Etiology, clinical management, and outcome after 3 mo were assessed. Results and limitations A causative pathogen, predominantly Escherichia coli (56%), was identified in 132 antibiotic-naive patients (88%) and 44 pretreated patients (51%); 16S rDNA analysis increased the detection rate by 10%. STIs were present in 34 cases (14%) (25 patients with Chlamydia trachomatis) and were not restricted to a specific age group. Enteroviruses were found in only two patients (1%). In naive patients, cultured bacteria were susceptible to fluoroquinolones and group 3 cephalosporins in >85% of cases (preateted patients: 42% and 67%, respectively). Primary empirical therapy was continued in 88% of naive patients for 11 d and in 77% of pretreated patients for 13 d with indwelling urinary catheters, rendering patients as high risk for switching. Only six patients (2.5%) underwent semicastration. Prostate-specific antigen levels halved within 3 mo, except in patients who were antibiotic naive and without detected pathogens. Study limitations included a lack of susceptibility testing in cases of STIs. Conclusions Even in antimicrobially pretreated patients, acute epididymitis is mainly of bacterial origin. STIs are not limited to patients aged <35 yr. Viral epididymitis seems a rare condition. Current guideline recommendations on empirical antimicrobial therapy are adequate. Patient summary Patients with acute epididymitis should receive appropriate diagnostics and antimicrobial therapy for safe conservative management. © 2014 European Association of Urology.

Walter J.,Robert Koch InstituteBerlin | Haller S.,Robert Koch InstituteBerlin | Blank H.,Robert Koch InstituteBerlin | Eckmanns T.,Robert Koch InstituteBerlin | And 2 more authors.
Eurosurveillance | Year: 2015

Voluntary surveillance systems in Germany suggest a recent decline in the incidence of infections (subsequent to at least 2010) with meticillin-resistant Staphylococcus aureus (MRSA) from various types of specimens and settings. We asked whether this decline is reflected by data from the mandatory national surveillance system for invasive MRSA infections. Our analysis is based on the population in Germany in 2010 to 2014. Cases were identified from passive reporting by microbiological laboratories of the diagnosis of MRSA from blood culture or cerebrospinal fluid. Respective clinical data were subsequently added to the notification. We calculated risk ratios (RR) between consecutive years, stratifying cases by sex, age and federal state of residence. The national incidence increased from 4.6 episodes per 100,000 persons in 2010 to 5.6 in 2012 (2011 vs 2010: RR: 1.13, 95% confidence interval (CI): 1.08–1.18; 2012 vs 2011: RR: 1.08, 95% CI: 1.04–1.13). It stagnated at 5.4 per 100,000 in 2013 (RR: 0.97, 95% CI: 0.93–1.01) before declining to 4.8 in 2014 (RR: 0.88, 95% CI: 0.84–0.91). This trend was observed in most, but not all federal states and strata of sex and age groups. Only 204 of 20,679 (1%) episodes of infection were notified as belonging to an outbreak. Our analysis corroborate previous findings that the incidence of invasive MRSA infections in Germany may be declining. © 2015, European Centre for Disease Prevention and Control (ECDC). All rights reserved.

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