A Gammaherpesviral Internal Repeat Contributes to Latency Amplification

BACKGROUND: Gammaherpesviruses cause important infections of humans, in particular in immunocompromised patients. The genomes of gammaherpesviruses contain variable numbers of internal repeats whose precise role for in vivo pathogenesis is not well understood. METHODOLOGY/PRINCIPAL FINDINGS: We used infection of laboratory mice with murine gammaherpesvirus 68 (MHV-68) to explore the biological role of the 40 bp internal repeat of MHV-68. We constructed several mutant viruses partially or completely lacking this repeat. Both in vitro and in vivo, the loss of the repeat did not substantially affect lytic replication of the mutant viruses. However, the extent of splenomegaly, which is associated with the establishment of latency, and the number of ex vivo reactivating and genome positive splenocytes were reduced. Since the 40 bp repeat is part of the hypothetical open reading frame (ORF) M6, it might function as part of M6 or as an independent structure. To differentiate between these two possibilities, we constructed an N-terminal M6STOP mutant, leaving the repeat structure intact but rendering ORF M6 unfunctional. Disruption of ORF M6 did neither affect lytic nor latent infection. In contrast to the situation in lytically infected NIH3T3 cells, the expression of the latency-associated genes K3 and ORF72 was reduced in the latently infected murine B cell line Ag8 in the absence of the 40 bp repeat. CONCLUSIONS/SIGNIFICANCE: These data suggest that the 40 bp repeat contributes to latency amplification and might be involved in the regulation of viral gene expression

Biografieforschung: theoretische Perspektiven und methodologische Konzepte für eine re-konstruktive Geschlechterforschung

Die Biografieforschung bezeichnet einen komplexen Forschungsansatz, der auf eine lange Geschichte des wissenschaftlichen Interesses an "persönlichen Dokumenten" verweisen kann. Sie ist eine voraussetzungsvolle Forschungsperspektive, die sich in zentralen Aspekten ihres Vorgehens auf Biografien als theoretisches Konzept, als historisch-empirischen Gegenstand und als komplexe methodologische Strategie bezieht. Andere Begriffe, welche oftmals synonym gebraucht, in der Biografieforschung aber systematisch unterschieden werden, sind "Lebensgeschichte" und "Lebenslauf". Die Autorin skizziert die Perspektiven einer rekonstruktiven Geschlechterforschung innerhalb der Biografieforschung, wozu sie auf die Differenzierungen empirischer Forschung, die methodologischen Prinzipien sowie auf Datenerhebung und Datenanalyse eingeht. Sie hebt insbesondere drei Kontextrelationen bei der Interpretation eines biografischen Textes hervor: Biografie, Interaktion, kulturelle Muster und soziale Regeln. Das skizzierte Konzept von Biografieforschung begreift sie als ein offenes Programm, das vielfältige Anknüpfungspunkte zu aktuellen theoretischen Diskussionen in der Geschlechterforschung aufweist. (ICI2

Analysis of the diagnostic criteria for Creutzfeldt-Jakob disease in Germany

Die Creutzfeldt-Jakob-Erkrankung (CJK) gehört zum Formenkreis der Prionerkrankungen. Bei einer durchschnittlichen Inzidenz für eine Prionerkrankung von 1,5 Fällen pro 1.000.0000 Einwohner erkranken jährlich ca. 120 Menschen an der sporadischen Form einer CJK (97% aller Prionerkrankungen). Die Patienten leiden an einer rasch fortschreitenden Demenz und Bewegungsstörung mit infauster Prognose. Da es keine kurative Therapie für die CJK gibt, ist eine zuverlässige Diagnose von grundlegender Bedeutung. Die vorliegende Arbeit analysiert die Diagnosekriterien in Deutschland (Stand 2012) in einer retrospektiven Fall-Kontroll Untersuchung im Hinblick auf Sensitivität und Spezifität der einzelnen klinischen und apparativen diagnostischen Kriterien sowie der Diagnosekriterien im Ganzen. Limitationen der Studie wurden aufgezeigt und die Auswirkungen auf die vorliegende Untersuchung wurden umfassend diskutiert. In der Analyse konnten Schwächen der Diagnosekriterien zum Zeitpunkt der Liquorpunktion (Sensitivität von 75%, Spezifität von 69%) aufgezeigt werden. Auf Grundlage der Ergebnisse wurden Verbesserungsmöglichkeiten mittels einer logistischen Regressionsanalyse eruiert, auch unter Einbeziehung der Real-Time Quaking-Induced Conversion (RT-QuIC) als ein neueres Testverfahren. Die Analyse ergab, dass durch Ergänzung des RT-QuIC und weitere Modifikationen eine Verbesserung der Diagnosekriterien auf 92% Sensitivität und 84% Spezifität erreicht werden kann. Dadurch wird die Diagnostik der CJK vereinfacht und verlässlicher gemacht werden kann.The Creutzfeldt-Jakob disease (CJD) is the most common prion disease. With an incident of 1.5 cases per 1,000,0000 inhabitants in Germany around 120 people (97% of all prion disease cases) are diagnosed with this disease every year. Patients suffer from a rapid progressiv dementia and movment disorder with poor prognosis. As there is no cure for the CJK a reliable diagnosis is fundamental. Therefore the present work analyzes the diagnostic criteria in Germany (as of 2012) in a retrospective case-controll study with regard to the sensitivity and specificity of the individual clinical and apparatus-based diagnostic criteria as well as the diagnostic criteria as a whole. Limitations of the study were identified and effects on the present study were discussed. The analysis revealed weaknesses in the diagnostic criteria at the time of the CSF puncture (sensitivity of 75%, specificity of 69%). On the basis of the results, possible improvement were determined by means of a logistic regression analysis, also taking into account Real-Time Quaking-Induced Conversion (RT-QuIC) as a more recent test procedure. The analysis showed that by supplementing the RT-QuIC and further modifications, a improvement of the diagnostic criteria to 92 % sensitivity and 84% specificity can be achieved.2021-12-0

Stroke Angel: Effect of Telemedical Prenotification on In-Hospital Delays and Systemic Thrombolysis in Acute Stroke Patients

Introduction: Door-to-CT scan time (DCT) and door-to-needle time (DNT) are important process measures in acute ischemic stroke (AIS) patients undergoing intravenous thrombolysis (IVT). We examined the impact of a telemedical prenotification by emergency medical service (EMS) (called the “Stroke Angel” program) on DCT and DNT and IVT rate compared to standard of care. Patients and Methods: Two prospective observational studies including AIS patients admitted via EMS from 2011 to 2013 (cohort I; n = 496) and from January 1, 2015 to May 31, 2018 (cohort II; n = 349) were conducted. After cohort I, the 4-Item Stroke Scale and a digital thrombolysis protocol were added. Multivariable logistic and linear regression analysis was performed. Results: In cohort I, DCT was lower in the intervention group (13 vs. 26 min using standard of care; p < 0.001), but no significant difference in median DNT (35 vs. 39 min; p = 0.24) was observed. In cohort II, a reduction of DCT (8 vs. 15 min; p < 0.001) and DNT (25 vs. 29 min p = 0.003) was observed in the intervention group. Compared to standard of care, the likelihood of DCT ≤10 min or DNT ≤20 min in the intervention group was 2.7 (adjusted odds ratio [aOR] 2.7; 95% CI: 2.1–3.5) and 1.8 (aOR 1.8; 95% CI: 1.1–2.9), respectively. In cohort II, IVT rate was higher (aOR 1.4; 95% CI: 1.1–1.9) in the intervention group. Conclusion: Although the positive effects of Stroke Angel in AIS provided a rationale for implementation in routine care, larger studies of practice implementation will be needed. Using Stroke Angel in the prehospital management of AIS impacts on important process measures of IVT delivery

The project "GND for cultural data" (GND4C)

Die zunehmende Präsenz von Museen, Archiven, Forschungs- und anderen Kulturgut verwahrenden Einrichtungen mit ihren Sammlungsobjekten im Internet verstärkt das Bedürfnis nach Vernetzung der Daten. Eine Voraussetzung für die semantische Verknüpfung von Datensätzen sind gemeinsam verwendete Normdaten. Die Gemeinsame Normdatei (GND), geführt von der Deutschen Nationalbibliothek, ist ein weithin anerkanntes Vokabular für die Beschreibung und das Information­Retrieval in der Bibliothekswelt. Veröffentlicht als Linked Open Data, unterstützt die GND semantische Interoperabilität und Nachnutzung von Daten. Objekte aus verschiedenen Häusern können zusammen aufgefunden werden und disziplinenübergreifende Forschung wird erleichtert. Doch um die GND sparten­ und fächerübergreifend öffnen zu können, muss sie an die neuen Anforderungen angepasst und aktiv zwischen den Communities geteilt werden. In den vier Teilen des Beitrags geben Mitarbeiterinnen und Mitarbeiter des DFG­geförderten Projekts „GND für Kulturdaten“ (GND4C) einen Einblick in den Stand der Untersuchungen, welche konzeptuellen Veränderungen erforderlich sein werden und wo sich die GND bereits heute als flexibel genug für die Wünsche der neuen Anwendungen erweist. Der erste Teil des Beitrags erläutert Ziel und Vorgehensweise des Projektes in vier thematischen Schwerpunkten: Organisationsstruktur des GND­Netzwerks, Datenmodell und Regelwerk der GND, technische Infrastruktur sowie Erfordernisse eines Dialogs über die Grenzen der Bibliothekswelt hinaus. Der zweite Teil beschreibt die Bedingungen und Erfordernisse für die Integration von Normdaten für Personen und Sachbegriffe in die GND, soweit diese nicht durch die Erschließungsarbeit der Bibliotheken vorgegeben sind. Daraufhin werden die Möglichkeiten des bestehenden Datenmodells und der bisher geltenden Regeln hinsichtlich ihres Anpassungspotenzials für historische Geografika sowie Bauwerke untersucht. Als ortsgebundene Entitäten haben Bauwerke Eigenschaften mit natürlichen Geografika gemeinsam, teilen als künstliche Objekte aber auch Eigenschaften der von Menschen geschaffenen Werke. Diese Dualität ortsgebundender Objekte gilt es, in der GND darstellbar zu machen. Im letzten Beitragsteil wird erläutert, warum es nur einen geringen Bedarf an einer konzeptionellen Änderung des Basisdatenmodells der GND gibt. Allerdings werden auch spezifische Eigenschaften identifiziert, die für Objekte aus Museen oder Archiven relevant sind und nicht vom aktuellen GND­Datenmodell abgedeckt werden. Diese zusätzlichen Anforderungen werden durch die Spezifikation von CORE­ und PLUS­Bereichen erfüllt. Das CORE­PLUS­Modell unterscheidet zwischen Minimalanforderungen an einen Normdatensatz in einem CORE­Modell und zusätzlichen Regeln für einen jeweils community­spezifischen PLUS­Bereich. Ausblick und Fazit zeigen die noch offenen Fragen auf, deren Lösung in der zweiten Hälfte der Projektlaufzeit ansteht.The growing online presence of museums, archives, research and other cultural heritage institutions requires efficient ways to semantically interlink the collection assets of our cultural treasures. One precondition for interlinking datasets is the shared use of authority files and controlled vocabularies. The Integrated Authority File (GND), managed by the German National Library, is a widely recognized vocabulary for description and information retrieval in library communities. Published as linked open data, the GND fosters semantic interoperability and re­use of data. Objects kept in different collections can be retrieved together, and cross­disciplinary research is facilitated. However, in order to open the GND for interdisciplinary and cross­domain use, it needs to be adapted and actively shared among the communities. In four parts of this article, members of the publicly funded project “GND for cultural data” (GND4C) give an insight into the state of the work, focusing on the question if, and to what extent the GND must adapt to the new requirements. The first part of this article outlines the objectives and approaches of the project according to its four focal points: the organizational structure of the GND network, the data model and rule set for the GND, the technical infrastructure as well as the requirements for community-building and communication beyond the library community. The second part describes the conditions and requirements for incorporating authority data for persons and concepts into the GND, as far as they are not predefined by the cataloging needs of libraries. The third part of this article looks at the expressiveness of the existing data model and rule set with regard to the cataloging of historic geographical entities as well as buildings and other human­made structures. Being immovable, buildings share properties with natural geographical entities. As artificial objects, they have properties in common with human­made works of any kind. Representing this dual nature of location­bound objects is one of the challenges for modelling. In the last part the authors outline why there is little need for conceptual changes to the basic GND data model. However, they identify specific properties pertinent to museum and archival objects which are not yet covered by the current model. These additional demands are met by specifying CORE and PLUS rules. The CORE­PLUS model distinguishes between minimum rules to define an authority entity in the CORE model, and additional rules in the particular community­specific PLUS rule set. The conclusion gives an overview of the issues still to be resolved in the second half of the GND4C­project

Latent Membrane Protein 1 of Epstein-Barr Virus Induces CD83 by the NF-κB Signaling Pathway

Epstein-Barr virus (EBV) infects human resting B cells and transforms them in vitro into continuously growing lymphoblastoid cell lines (LCLs). EBV nuclear antigen 2 (EBNA2) is one of the first viral proteins expressed after infection. It is able to transactivate viral as well as cellular target genes by interaction with cellular transcription factors. EBNA2 target genes can be studied easily by using an LCL (ER/EB2-5) in which wild-type EBNA2 is replaced by an estrogen-inducible EBNA2. Since the cell surface molecule CD83, a member of the immunoglobulin superfamily and a marker for mature dendritic cells, appeared on the surface of ER/EB2-5 cells within 3 h after the addition of estrogen, we analyzed the regulation of CD83 induction by EBV in more detail. Despite its rapid induction, CD83 turned out to be an indirect target gene of EBNA2. We could show that the viral latent membrane protein 1 (LMP1) is responsible for the induction of CD83 by using an LCL expressing a ligand- or antibody-inducible recombinant nerve growth factor receptor-LMP1 fusion protein. The inducibility of the CD83 promoter by LMP1 was mediated by the activation of NF-κB, as seen by use of luciferase reporter assays using the CD83 promoter and LMP1 mutants. Additionally, fusion constructs of the transmembrane domain of LMP1 and the intracellular signaling domain of CD40, TNF-R1, and TNF-R2 likewise transactivated the CD83 promoter via NF-κB. Our studies show that CD83 is also a target of the NF-κB signaling pathway in B cells

Characterization of ACE inhibitors and AT1R antagonists with regard to their effect on ACE2 expression and infection with SARS-CoV-2 using a Caco-2 cell model

Blood-pressure-lowering drugs are proposed to foster SARS-CoV-2 infection by pharmacological upregulation of angiotensin-converting enzyme 2 (ACE2), the binding partner of the virus spike (S) protein, located on the surface of the host cells. Conversely, it is postulated that angiotensin–renin system antagonists may prevent lung damage caused by SARS-CoV-2 infection, by reducing angiotensin II levels, which can induce permeability of lung endothelial barrier via its interaction with the AT1 receptor (AT1R). Methods: We have investigated the influence of the ACE inhibitors (lisinopril, captopril) and the AT1 antagonists (telmisartan, olmesartan) on the level of ACE2 mRNA and protein expression as well as their influence on the cytopathic effect of SARS-CoV-2 and on the cell barrier integrity in a Caco-2 cell model. Results: The drugs revealed no effect on ACE2 mRNA and protein expression. ACE inhibitors and AT1R antagonist olmesartan did not influence the infection rate of SARS-CoV-2 and were unable to prevent the SARS-CoV-2-induced cell barrier disturbance. A concentration of 25 µg/mL telmisartan significantly reduced the virus replication rate. Conclusion: ACE inhibitors and AT1R antagonist showed neither beneficial nor detrimental effects on SARS-CoV-2-infection and cell barrier integrity in vitro at pharmacologically relevant concentrations

Nicotine delivery and relief of craving after consumption of European JUUL e-cigarettes prior and after pod modification

The emergence of e-cigarettes on the consumer market led to a tremendous rise in e-cigarette consumption among adolescents in the United States. The success of JUUL and other pod systems was linked to its high nicotine delivery capacity. In compliance with the European Tobacco Product directive, liquid nicotine contents in the European JUUL variants are limited to 20 mg/mL or below. A short time after launching the initial version in Europe, JUUL pods have been modified in terms of the wick material used. This modification has been demonstrated previously to lead to an elevated aerosol generation, consequently, to a larger amount of nicotine per puff generated. The present study was designed to assess whether the mentioned differences between the “initial” and “modified” JUUL versions may cause a significant difference during consumption, and how nicotine delivery compares with tobacco cigarettes. In this single-center three-arm study, nicotine pharmacokinetics and influence on urge to smoke/vape were compared for tobacco cigarettes, the “initial” version of the European JUUL, and the “modified” version of the European JUUL. Participants, 15 active smokers and 17 active e-cigarette users, were instructed to consume their study product according to a pre-directed puffing protocol. Venous blood was sampled for nicotine analysis to cover the acute phase and the first 30 min after starting. Nicotine delivery and the reduction of urge to smoke/vape upon usage of both European JUUL variants were lower in comparison to tobacco cigarettes. This suggests a lower addictive potential. Modification of the pod design did not result in significant differences at the first ten puffs, as confirmed by a vaping machine experiment. Apparently, the limitations by the initially used wick material only come into effect after longer usage time