Structure and Function Relationship of the Autotransport and Proteolytic Activity of EspP from Shiga Toxin-Producing Escherichia coli

BACKGROUND: The serine protease autotransporter EspP is a proposed virulence factor of Shiga toxin-producing Escherichia coli (STEC). We recently distinguished four EspP subtypes (EspPalpha, EspPbeta, EspPgamma, and EspPdelta), which display large differences in transport and proteolytic activities and differ widely concerning their distribution within the STEC population. The mechanisms underlying these functional variations in EspP subtypes are, however, unknown. METHODOLOGY/PRINCIPAL FINDINGS: The structural basis of proteolytic and autotransport activity was investigated using transposon-based linker scanning mutagenesis, site-directed mutagenesis and structure-function analysis derived from homology modelling of the EspP passenger domain. Transposon mutagenesis of the passenger domain inactivated autotransport when pentapeptide linker insertions occurred in regions essential for overall correct folding or in a loop protruding from the beta-helical core. Loss of proteolytic function was limited to mutations in Domain 1 in the N-terminal third of the EspP passenger. Site-directed mutagenesis demonstrated that His(127), Asp(156) and Ser(263) in Domain 1 form the catalytic triad of EspP. CONCLUSIONS/SIGNIFICANCE: Our data indicate that in EspP i) the correct formation of the tertiary structure of the passenger domain is essential for efficient autotransport, and ii) an elastase-like serine protease domain in the N-terminal Domain 1 is responsible for the proteolytic phenotype. Lack of stabilizing interactions of Domain 1 with the core structure of the passenger domain ablates proteolytic activity in subtypes EspPbeta and EspPdelta

Unravelling the Stability of Nightingale Song Over Time and Space Using Open, Citizen Science and Shared Data

Open science approaches enable and facilitate the investigation of many scientific questions in bioacoustics, such as studies on the temporal and spatial evolution of song, as in vocal dialects. In contrast to previous dialect studies, which mostly focused on songbird species with a small repertoire, here we studied the common nightingale (Luscinia megarhynchos), a bird species with a complex and large repertoire. To study dialects on the population level in this species, we used recordings from four datasets: an open museum archive, a citizen science platform, a citizen science project, and shared recordings from academic researchers. We conducted the to date largest temporal and geographic dialect study of birdsong including recordings from 1930 to 2019 and from 13 European countries, with a geographical coverage of 2,652 km of linear distance. To examine temporal stability and spatial dialects, a catalog of 1,868 song types of common nightingales was created. Instead of dialects, we found a high degree of stability over time and space in both, the sub-categories of song and in the occurrence of song types. For example, the second most common song type in our datasets occurred over nine decades and across Europe. In our case study, open and citizen science data proved to be equivalent, and in some cases even better, than data shared by an academic research group. Based on our results, we conclude that the combination of diverse and open datasets was particularly useful to study the evolution of song in a bird species with a large repertoire.Peer Reviewe

Unravelling the Stability of Nightingale Song Over Time and Space Using Open, Citizen Science and Shared Data

Open science approaches enable and facilitate the investigation of many scientific questions in bioacoustics, such as studies on the temporal and spatial evolution of song, as in vocal dialects. In contrast to previous dialect studies, which mostly focused on songbird species with a small repertoire, here we studied the common nightingale (Luscinia megarhynchos), a bird species with a complex and large repertoire. To study dialects on the population level in this species, we used recordings from four datasets: an open museum archive, a citizen science platform, a citizen science project, and shared recordings from academic researchers. We conducted the to date largest temporal and geographic dialect study of birdsong including recordings from 1930 to 2019 and from 13 European countries, with a geographical coverage of 2,652 km of linear distance. To examine temporal stability and spatial dialects, a catalog of 1,868 song types of common nightingales was created. Instead of dialects, we found a high degree of stability over time and space in both, the sub-categories of song and in the occurrence of song types. For example, the second most common song type in our datasets occurred over nine decades and across Europe. In our case study, open and citizen science data proved to be equivalent, and in some cases even better, than data shared by an academic research group. Based on our results, we conclude that the combination of diverse and open datasets was particularly useful to study the evolution of song in a bird species with a large repertoire

Community engagement and data quality: best practices and lessons learned from a citizen science project on birdsong

Citizen Science (CS) is a research approach that has become popular in recent years and offers innovative potential for dialect research in ornithology. As the scepticism about CS data is still widespread, we analysed the development of a 3-year CS project based on the song of the Common Nightingale (Luscinia megarhynchos) to share best practices and lessons learned. We focused on the data scope, individual engagement, spatial distribution and species misidentifications from recordings generated before (2018, 2019) and during the COVID-19 outbreak (2020) with a smartphone using the ‘Naturblick’ app. The number of nightingale song recordings and individual engagement increased steadily and peaked in the season during the pandemic. 13,991 nightingale song recordings were generated by anonymous (64%) and non-anonymous participants (36%). As the project developed, the spatial distribution of recordings expanded (from Berlin based to nationwide). The rates of species misidentifications were low, decreased in the course of the project (10–1%) and were mainly affected by vocal similarities with other bird species. This study further showed that community engagement and data quality were not directly affected by dissemination activities, but that the former was influenced by external factors and the latter benefited from the app. We conclude that CS projects using smartphone apps with an integrated pattern recognition algorithm are well suited to support bioacoustic research in ornithology. Based on our findings, we recommend setting up CS projects over the long term to build an engaged community which generates high data quality for robust scientific conclusions.Gesellschaftliches Engagement und Datenqualität: Bewährte Praktiken und Erfahrungen aus einem bürgerwissenschaftlichen Projekt zum Vogelgesang Citizen Science (CS) ist eine Forschungsmethode, die in den letzten Jahren an Bedeutung gewonnen hat und innovatives Potenzial für die Dialektforschung in der Ornithologie bietet. Da die Vorbehalte gegenüber CS-Daten immer noch weit verbreitet sind, haben wir die Entwicklung eines dreijährigen CS-Projekts zum Gesang der Nachtigall (Luscinia megarhynchos) analysiert, um bewährte Praktiken und gewonnene Erfahrungen darzustellen. Wir fokussierten uns auf den Datenumfang, das individuelle Engagement von Teilnehmenden, die räumliche Verteilung und die Fehlbestimmungen von Arten aus Aufnahmen, die vor (2018, 2019) und während des COVID-19-Ausbruchs (2020) mit einem Smartphone unter Verwendung der "Naturblick" App erstellt wurden. Die Anzahl der Aufnahmen von Nachtigallgesängen und das individuelle Engagement stiegen stetig an und erreichten ihren Höhepunkt in der Saison während der Pandemie. 13.991 Aufnahmen von Nachtigallgesängen wurden von anonymen (64%) und nicht-anonymen Teilnehmenden (36%) erstellt. Im Laufe des Projekts weitete sich die räumliche Verteilung der Aufnahmen aus (von Berlin auf bundesweit). Die Rate der Fehlbestimmungen war gering, ging im Laufe des Projekts zurück (von 10% auf 1%) und wurde hauptsächlich von gesanglichen Ähnlichkeiten mit anderen Vogelarten beeinflusst. Unsere Studie zeigte außerdem, dass das gesellschaftliche Engagement und die Datenqualität nicht direkt von den durchgeführten Disseminationsaktivitäten beeinflusst wurden, sondern dass erstere von externen Faktoren abhingen und letztere von der App profitierte. Wir schließen daraus, dass CS-Projekte, die Smartphone-Apps mit einem integrierten Mustererkennungsalgorithmus verwenden, gut geeignet sind, um die bioakustische Forschung in der Ornithologie zu unterstützen. Auf der Grundlage unserer Ergebnisse empfehlen wir, CS-Projekte langfristig zu etablieren, um eine aktive Teilnehmergemeinschaft (Community) aufzubauen, die qualitativ hochwertige Daten für fundierte wissenschaftliche Schlussfolgerungen generiert

HIV pre-exposure prophylaxis was associated with no impact on sexually transmitted infection prevalence in a high-prevalence population of predominantly men who have sex with men, Germany, 2018 to 2019

Introduction: Despite increased use of pre-exposure prophylaxis (PrEP) in Germany, HIV infection rates are not declining and little is known about how this prevention method affects the prevalence of sexually transmitted infections (STI) among men who have sex with men (MSM). Aim: We studied, in a large multicentre cohort, STI point prevalence, co-infection rates, anatomical location and influence of PrEP. Methods: The BRAHMS study was a prospective cohort study conducted at 10 sites in seven major German cities that enrolled MSM reporting increased sexual risk behaviour. At screening visits, MSM were tested for Mycoplasma genitalium (MG), Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT) and Treponema pallidum (TP), and given a behavioural questionnaire. With binomial regression, we estimated prevalence ratios (PR) and 95% confidence intervals (CI) for the association of PrEP and STI. Results: We screened 1,043 MSM in 2018 and 2019, with 53.0% currently using PrEP. At screening, 370 participants (35.5%) had an STI. The most common pathogen was MG in 198 (19.0%) participants, followed by CT (n = 133; 12.8%), NG (n = 105; 10.1%) and TP (n = 37; 3.5%). Among the 370 participants with at least one STI, 14.6% (n = 54) reported STI-related symptoms. Infection prevalence was highest at anorectal site (13.4% MG, 6.5% NG, 10.2% CT). PrEP use was not statistically significant in adjusted models for STI (PR: 1.10; 95% CI: 0.91–1.32), NG/CT, only NG or only CT. Conclusions: Prevalence of asymptomatic STI was high, and PrEP use did not influence STI prevalence in MSM eligible for PrEP according to national guidelines.Peer Reviewe

German evidence and consensus‐based (S3) guideline: Vaccination recommendations for the prevention of HPV‐associated lesions

Anogenital and oropharyngeal infections with human papilloma viruses (HPV) are common. Clinically manifest disease may significantly impact quality of life; the treatment of HPV-associated lesions is associated with a high rate of recurrence and invasive neoplasms, such as cervical, anal, vulvar, penile, and oropharyngeal cancers, which are characterized by significant morbidity and mortality. Vaccination against HPV is an effective and safe measure for the primary prevention of HPV-associated lesions, but immunization rates are still low in Germany. The present publication is an abridged version of the German evidence and consensus-based guideline "Vaccination recommendations for the prevention of HPV-associated lesions", which is available on the website of the German Association of the Scientific Medical Societies (AWMF). On the basis of a systematic review with meta-analyses, a representative panel developed and agreed upon recommendations for the vaccination of different populations against HPV. In addition, consensus-based recommendations were developed for specific issues relevant to everyday practice. Based on current evidence and a representative expert consensus, these recommendations are intended to provide guidance in a field in which there is often uncertainty and in which both patients and health care providers are sometimes confronted with controversial and emotionally charged points of view

Long-term Mortality in HIV-Positive Individuals Virally Suppressed for >3 Years With Incomplete CD4 Recovery

Virally suppressed HIV-positive individuals on combination antiretroviral therapy who do not achieve a CD4 count >200 cells/µL have substantially increased long-term mortality. The increased mortality was seen across different patient groups and for all causes of deat

Meat Authentication: A New HPLC–MS/MS Based Method for the Fast and Sensitive Detection of Horse and Pork in Highly Processed Food

Fraudulent blending of food products with meat from undeclared species is a problem on a global scale, as exemplified by the European horse meat scandal in 2013. Routinely used methods such as ELISA and PCR can suffer from limited sensitivity or specificity when processed food samples are analyzed. In this study, we have developed an optimized method for the detection of horse and pork in different processed food matrices using MRM and MRM³ detection of species-specific tryptic marker peptides. Identified marker peptides were sufficiently stable to resist thermal processing of different meat products and thus allow the sensitive and specific detection of pork or horse in processed food down to 0.24% in a beef matrix system. In addition, we were able to establish a rapid 2-min extraction protocol for the efficient protein extraction from processed food using high molar urea and thiourea buffers. Together, we present here the specific and sensitive detection of horse and pork meat in different processed food matrices using MRM-based detection of marker peptides. Notably, prefractionation of proteins using 2D-PAGE or off-gel fractionation is not necessary. The presented method is therefore easily applicable in analytical routine laboratories without dedicated proteomics background

Antiretrovirale Therapie der HIV-Infektion

Die antiretrovirale Therapie der HIV-Infektion hat sich in den letzten zwei Jahren aufgrund neuer Therapieoptionen und -strategien deutlich gewandelt. Die Empfehlungen zur antiretroviralen Therapie wurden daher in Abhängigkeit von klinischen, immunologischen und virologischen Parametern von einem deutsch-österreichischen Expertengremium überarbeitet. Eingebrachte Studienergebnisse betreffen insbesondere die Wertung der Kombinationstherapien von zwei nukleosidanalogen reverse Transkriptase-Inhibitoren (NRTI) in Kombination mit einem nichtnukleosidanalogen reverse Transkriptase-Inhibitor (NNRTI) oder einem Proteaseinhibitor und mit drei NRTI. Diese Kombinationen werden hinsichtlich ihrer therapeutischen Effektivität und ihres Nebenwirkungsprofils gewichtet. Für eine effektive Therapie sind eine Resistenztestung und eine Bestimmung von Plasmamedikamentenspiegeln bedeutsame Parameter, die für das Monitoring wichtig sind, sowie Faktoren, die einen Wechsel und eine Unterbrechung der Therapie indizieren, werden aufgezeigt.Antiretroviral Therapy in HIV-Infections During the last two years, antiretroviral therapy of HIV infections has changed dramatically. Therefore a board of German and Austrian physicians experienced in antiretroviral research and HIV patient care revised the guidelines for antiretroviral therapy and treatment and have strongly been influenced by data from latest studies on clinical, immunological and viral parameters. These studies mainly investigated combination therapies such as two nucleoside reverse transcritase inhibitor (NRTI) with one protease inhibitor or one non-nucleoside reverse transcriptase inhibitor with three NRTI. These combination therapies are evaluated in regard to their therapeutic effectiveness and their side effects. Resistance testing and drug monitoring are most important during therapy. Parameters which necessitate a change in or interruption of therapy are discussed