The role of the human RNase H2 for the mobilization of non-LTR-retrotransposons

Im Rahmen der vorliegenden Dissertation wurden mit Hilfe des CRISPR-Cas9-Systems monoklonale RNase H2-defiziente, humane Zelllinien generiert. Diese wiesen signifikante Proliferationsdefizite im Vergleich zu ihren Parentalzellen auf, was auf eine Arretierung des Zellzyklus in der G2-Phase zurückzuführen war. Dieses erste Anzeichen für das Vorliegen von DNA-Schäden konnte unter Verwendung verschiedener Methoden bestätigt werden. So waren RNASEH2A-defiziente Zellen von vermehrten Strangbrüchen, cytoplasmatischen DNA-haltigen Strukturen (sog. Mikronuklei) sowie höheren Mutationsraten betroffen, was als Ausdruck für genomische Instabilität gedeutet wurde. Des Weiteren gab es Hinweise auf eine leicht vermehrte Inkorporation von Ribonukleotiden in die genomische DNA der RNase H2-knockout-Zellen. Darüber hinaus konnte in dieser Arbeit erstmalig festgestellt werden, dass die RNase H2 einen positiven Einfluss auf die Mobilisierung von non-LTR-Retrotransposonen ausübt.Within the framework of this dissertation, monoclonal RNase H2-deficient human cell lines were generated by means of the CRISPR-Cas9 system. They featured a significant proliferative deficit compared to their parental cells that was ascribed to a G2-dependent cell cycle arrest. This first hint towards the existence of DNA-damage could subsequently be confirmed by the use of various techniques. The RNASEH2A-deficient cells were affected by increased numbers of strand breaks as well as cytoplasmic DNA-containing structures called micronuclei plus higher mutation rates, which was interpreted as a signal for elevated genomic instability. In addition to that, a lightly increased incorporation rate of ribonucleotides within the genomic DNA of RNase H2 knockout cells could be detected.Beyond that, it could be proved for the first time within this work that the RNase H2 exerts a positive influence on the mobilisation of non-LTR-retrotransposons

Mittel- und langfristige Finanzierung der Pflegevorsorge

Das österreichische Modell der Pflegevorsorge steht durch den demografischen Wandel sowie die verstärkte Individualisierung der Gesellschaft vor einer immer dringlicher werdenden Herausforderung. Einerseits ist durch die steigende Alterung der Bevölkerung eine wachsende Belastung der öffentlichen Budgets zu erwarten. Andererseits kann durch die steigende Erwerbsbeteiligung von Frauen sowie aufgrund des Rückgangs der Kinderanzahl eine Erhöhung der Nachfrage nach außerfamiliären Pflegeleistungen erwartet werden. Pflegebedürftig zu werden, stellt ein Risiko dar, welches vor allem BezieherInnen von geringeren Pensionen finanziell überfordern kann. Die Daten über die soziale Struktur der PflegegeldbezieherInnen zeigen, dass das Pflegegeld vor allem an BezieherInnen niedriger Pensionen ausgezahlt wird. Diese Studie zielt auf die Analyse der Finanzströme der österreichischen Pflegevorsorge ab. Der erste Teil stellt eine Status-quo-Erhebung des österreichischen Pflegesystems dar. Dabei werden der Finanzierungsaufwand sowie die Finanzierungsstruktur der Langzeitpflege in Österreich erfasst. Der zweite Teil der Studie beschäftigt sich mit der mittel- und langfristigen Kostenentwicklung der Pflegevorsorge aufgrund der derzeit geltenden Regelungen. Im dritten und letzten Teil der Studie werden Pflegesysteme dreier europäischer Länder untersucht: Deutschland, Dänemark und die Niederlande

Alternative Finanzierungsformen der Pflegevorsorge

Die Studie diskutiert unterschiedliche Organisations- und Finanzierungsvarianten für die Pflegevorsorge in Österreich. Die Analyse der ökonomischen Wirkungen einer Steuer- versus einer Beitragslösung zeigt, dass sowohl die Verteilungswirkungen als auch die Wirkungen auf Wachstum und Beschäftigung eine Steuerlösung nahe legen. Bei einer unzureichenden Ergiebigkeit der Steuerfinanzierung wäre ein Mischsystem denkbar. Aus organisatorischer Sicht wird auf die Notwendigkeit einer weiteren Angleichung der unterschiedlichen Systeme in den Bundesländern hingewiesen. Die verwaltungstechnische Problematik der Heranziehung des Vermögens der zu Pflegenden könnte mit vermögensbezogenen Steuern ausgeglichen werden. Können sich Bund und Länder auf eine Vereinfachung der Organisation einigen und werden unterschiedliche Kanäle der Finanzierung herangezogen, bietet sich als organisatorischer Überbau für Geld- und Sachleistungen eine Fondslösung an, die hinsichtlich Ergiebigkeit, Volatilität, Verteilungseffekte und anderer ökonomischer Effekte optimiert und an sich ändernden Bedingungen angepasst werden kann

Umverteilung im Wohlfahrtsstaat

Mit der Zunahme der Ungleichheit in der Verteilung der Markteinkommen in den letzten Jahrzehnten wurden sowohl in den ökonomischen Analysen als auch in der wirtschaftspolitischen Diskussion Verteilungsfragen wieder aktuell. Dazu kommen in den nächsten Jahren die Folgen der tiefen Krise, die zum einen die Bevölkerung in unterschiedlichem Ausmaß trifft und zum anderen Fragen nach den Verteilungswirkungen der Steuern und Staatsausgaben neu aufwerfen wird, wenn die budgetären Folgekosten der Krisenbekämpfung zu finanzieren sein werden. Die vorliegende Studie untersucht die direkten Umverteilungswirkungen der Aktivitäten des öffentlichen Sektors auf die „Wohlfahrt“ der privaten Haushalte für die Jahre 2000 und 2005. Sie schließt an die bisherigen Studien des WIFO (Guger, 1987, 1996A) zu diesem Thema in möglichst vergleichbarer Form an.The increasing inequality in the distribution of market incomes in recent decades has made distribution issues topical again, both in economic analyses and in the discourse on economic policy. The consequences of the current severe economic crisis will further enliven the discussion: on the one hand, the crisis affects different sectors of the population to varying extents, and on the other hand it will again raise questions about the redistribution effects of taxes when it comes to financing the follow-up costs of combating the crisis. This study examines the direct redistributive effects of the activities of the public sector on the ‘welfare’ of private households in the years 2000 and 2005. It follows up the previous studies by the austrian institute for economic Research (wifo) on this theme (Guger, 1987, 1996a) in as comparable a form as possible

Mucus detachment by host metalloprotease meprin \beta requires shedding of its inactive pro-form, which is abrogated by the pathogenic protease RgpB

The host metalloprotease meprin β is required for mucin 2 (MUC2) cleavage, which drives intestinal mucus detachment and prevents bacterial overgrowth. To gain access to the cleavage site in MUC2, meprin β must be proteolytically shed from epithelial cells. Hence, regulation of meprin β shedding and activation is important for physiological and pathophysiological conditions. Here, we demonstrate that meprin β activation and shedding are mutually exclusive events. Employing ex vivo small intestinal organoid and cell culture experiments, we found that ADAM-mediated shedding is restricted to the inactive pro-form of meprin β and is completely inhibited upon its conversion to the active form at the cell surface. This strict regulation of meprin β activity can be overridden by pathogens, as demonstrated for the bacterial protease Arg-gingipain (RgpB). This secreted cysteine protease potently converts membrane-bound meprin β into its active form, impairing meprin β shedding and its function as a mucus-detaching protease

Bovine Herpesvirus Type 1 (BHV-1) UL49.5 Luminal Domain Residues 30 to 32 Are Critical for MHC-I Down-Regulation in Virus-Infected Cells

Bovine herpesvirus type 1 (BHV-1) UL49.5 inhibits transporter associated with antigen processing (TAP) and down-regulates cell-surface expression of major histocompatibility complex (MHC) class I molecules to promote immune evasion. We have constructed a BHV-1 UL49.5 cytoplasmic tail (CT) null and several UL49.5 luminal domain mutants in the backbone of wild-type BHV-1 or BHV-1 UL49.5 CT- null viruses and determined their relative TAP mediated peptide transport inhibition and MHC-1 down-regulation properties compared with BHV-1 wt. Based on our results, the UL49.5 luminal domain residues 30–32 and UL49.5 CT residues, together, promote efficient TAP inhibition and MHC-I down-regulation functions. In vitro, BHV-1 UL49.5 Δ30–32 CT-null virus growth property was similar to that of BHV-1 wt and like the wt UL49.5, the mutant UL49.5 was incorporated in the virion envelope and it formed a complex with gM in the infected cells

Epithelial RNase H2 Maintains Genome Integrity and Prevents Intestinal Tumorigenesis in Mice

BACKGROUND & AIMS: RNase H2 is a holoenzyme, composed of 3 subunits (ribonuclease H2 subunits A, B, and C), that cleaves RNA:DNA hybrids and removes mis-incorporated ribonucleotides from genomic DNA through ribonucleotide excision repair. Ribonucleotide incorporation by eukaryotic DNA polymerases occurs during every round of genome duplication and produces the most frequent type of naturally occurring DNA lesion. We investigated whether intestinal epithelial proliferation requires RNase H2 function and whether RNase H2 activity is disrupted during intestinal carcinogenesis. METHODS: We generated mice with epithelial-specific deletion of ribonuclease H2 subunit B (H2bΔIEC) and mice that also had deletion of tumor-suppressor protein p53 (H2b/p53ΔIEC); we compared phenotypes with those of littermate H2bfl/fl or H2b/p53fl/fl (control) mice at young and old ages. Intestinal tissues were collected and analyzed by histology. We isolated epithelial cells, generated intestinal organoids, and performed RNA sequence analyses. Mutation signatures of spontaneous tumors from H2b/p53ΔIEC mice were characterized by exome sequencing. We collected colorectal tumor specimens from 467 patients, measured levels of ribonuclease H2 subunit B, and associated these with patient survival times and transcriptome data. RESULTS: The H2bΔIEC mice had DNA damage to intestinal epithelial cells and proliferative exhaustion of the intestinal stem cell compartment compared with controls and H2b/p53ΔIEC mice. However, H2b/p53ΔIEC mice spontaneously developed small intestine and colon carcinomas. DNA from these tumors contained T>G base substitutions at GTG trinucleotides. Analyses of transcriptomes of human colorectal tumors associated lower levels of RNase H2 with shorter survival times. CONCLUSIONS: In analyses of mice with disruption of the ribonuclease H2 subunit B gene and colorectal tumors from patients, we provide evidence that RNase H2 functions as a colorectal tumor suppressor. H2b/p53ΔIEC mice can be used to study the roles of RNase H2 in tissue-specific carcinogenesis

The Priority position paper: protecting Europe's food chain from prions

International audienceBovine spongiform encephalopathy (BSE) created a global European crisis in the 1980s and 90s, with very serious health and economic implications. Classical BSE now appears to be under control, to a great extent as a result of a global research effort that identified the sources of prions in meat and bone meal (MBM) and developed new animal-testing tools that guided policy. Priority ( www.prionpriority.eu ) was a European Union (EU) Framework Program 7 (FP7)-funded project through which 21 European research institutions and small and medium enterprises (SMEs) joined efforts between 2009 and 2014, to conduct coordinated basic and applied research on prions and prion diseases. At the end of the project, the Priority consortium drafted a position paper ( www.prionpriority.eu/Priority position paper) with its main conclusions. In the present opinion paper, we summarize these conclusions. With respect to the issue of re-introducing ruminant protein into the feed-chain, our opinion is that sustaining an absolute ban on feeding ruminant protein to ruminants is essential. In particular, the spread and impact of non-classical forms of scrapie and BSE in ruminants is not fully understood and the risks cannot be estimated. Atypical prion agents will probably continue to represent the dominant form of prion diseases in the near future in Europe. Atypical L-type BSE has clear zoonotic potential, as demonstrated in experimental models. Similarly, there are now data indicating that the atypical scrapie agent can cross various species barriers. More epidemiological data from large cohorts are necessary to reach any conclusion on the impact of its transmissibility on public health. Re-evaluations of safety precautions may become necessary depending on the outcome of these studies. Intensified searching for molecular determinants of the species barrier is recommended, since this barrier is key for important policy areas and risk assessment. Understanding the structural basis for strains and the basis for adaptation of a strain to a new host will require continued fundamental research, also needed to understand mechanisms of prion transmission, replication and how they cause nervous system dysfunction and death. Early detection of prion infection, ideally at a preclinical stage, also remains crucial for development of effective treatment strategies

Community-acquired pneumonia related to intracellular pathogens

Community-acquired pneumonia (CAP) is associated with high rates of morbidity and mortality worldwide; the annual incidence of CAP among adults in Europe has ranged from 1.5 to 1.7 per 1000 population. Intracellular bacteria are common causes of CAP. However, there is considerable variation in the reported incidence between countries and change over time. The intracellular pathogens that are well established as causes of pneumonia are Legionella pneumophila, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Chlamydophila psittaci, and Coxiella burnetii. Since it is known that antibiotic treatment for severe CAP is empiric and includes coverage of typical and atypical pathogens, microbiological diagnosis bears an important relationship to prognosis of pneumonia. Factors such as adequacy of initial antibiotic or early de-escalation of therapy are important variables associated with outcomes, especially in severe cases. Intracellular pathogens sometimes appear to cause more severe disease with respiratory failure and multisystem dysfunction associated with fatal outcomes. The clinical relevance of intracellular pathogens in severe CAP has not been specifically investigated. We review the prevalence, general characteristics, and outcomes of severe CAP cases caused by intracellular pathogens

Between East and West: Niederösterreichs adelige Grundherrschaft 1550-1750

The Jack of a clearly discernible border between the regions of Grund- and Gutsherrschaft as well as the actual overlapping of the two systems in East-Central Europe has necessitated a reassessment of regional differentiation and the postulation of transitional types of manorial systems. Pointing towards the fact, that the main characteristics of „second serfdom" do not apply to the regions of present day Austria, the author selects as his foremost criterion for the analysis of the position of the Lower and Upper Austrian variants of the manorial system the varying amount of the labour rent extracted by the landlord. The comparison of the two Austrian provinces based on material from the mid-eighteenth century shows that labour dues were of major importance only in a few regions at the eastern fringe of the provinces, without ever reaching the dimensions characteristic for the core zones of Gutsherrschaft east of the river Elbe and for Bohemia after the Thirty Years' War. On the other hand, the picture of manorial estates furnished with agrarian and industrial demesnes, monopolies as weil as rents in cash and kind differs so significantly from the ideal type of Grundherrschaft that the introduction of a transitional category, like Alfred Hoffmann's concept of „Wirtschaftsherrschaft", seems to be justified, although this system was neither limited to the Austrian lands nor was it systematically applied throughout them.The Jack of a clearly discernible border between the regions of Grund- and Gutsherrschaft as well as the actual overlapping of the two systems in East-Central Europe has necessitated a reassessment of regional differentiation and the postulation of transitional types of manorial systems. Pointing towards the fact, that the main characteristics of „second serfdom" do not apply to the regions of present day Austria, the author selects as his foremost criterion for the analysis of the position of the Lower and Upper Austrian variants of the manorial system the varying amount of the labour rent extracted by the landlord. The comparison of the two Austrian provinces based on material from the mid-eighteenth century shows that labour dues were of major importance only in a few regions at the eastern fringe of the provinces, without ever reaching the dimensions characteristic for the core zones of Gutsherrschaft east of the river Elbe and for Bohemia after the Thirty Years' War. On the other hand, the picture of manorial estates furnished with agrarian and industrial demesnes, monopolies as weil as rents in cash and kind differs so significantly from the ideal type of Grundherrschaft that the introduction of a transitional category, like Alfred Hoffmann's concept of „Wirtschaftsherrschaft", seems to be justified, although this system was neither limited to the Austrian lands nor was it systematically applied throughout them