Etablierung eines revers–genetischen Systems für feline Coronaviren

Im Rahmen dieser Arbeit erfolgte die Etablierung eines revers-genetischen Systems für ein Serotyp I felines Coronavirus (FCoV). Dabei wurden folgende Ergebnisse erzielt: 1) Als Grundlage für die Etablierung des Systems wurde die komplette Genomsequenz des FCoV Stammes Black bestimmt und anhand dieser Sequenz der Genomaufbau ermittelt. Es ließ sich der für Coronaviren typische Genomaufbau nachweisen. 2) Die Herstellung eines infektiösen Klons beinhaltete die Integration einer kompletten cDNA Kopie des FCoV Stammes Black in das Vaccinia Virus Genom, welches als Vektor diente. Die coronavirale cDNA wurde in vitro transkribiert und die synthetische RNA in Zellen elektroporiert. Rekombinante FCoV ließen sich nachweisen und somit war die synthetische RNA infektiös, also in der Lage, den coronaviralen Replikationszyklus zu initiieren und zu durchlaufen. Die Analyse der gewonnenen rekombinanten Viren erfolgte nach Infektion feliner Zellkulturen mit verschiedenen Ansätzen. Dabei konnte festgestellt werden, dass die rekombinanten Viren die gleichen Eigenschaften wie der ursprüngliche FCoV Stamm Black besitzen. 3) In nachfolgenden Versuchen erfolgte die Herstellung zweier Reportergenexprimierender FCoV mit dem etablierten revers-genetischen System. Dabei wurden im FCoV Stamm Black Genom die akzessorischen Gene 3abc durch das „green fluorescent protein“ (GFP) oder Renilla Luciferase ersetzt. Die stabile Expression der Reportergene konnte in felinen Zellkulturen nachgewiesen werden. 4) Feline Monozyten, Makrophagen und dendritische Zellen (DCs), die als Zielzellen sowohl von FCoV als auch von anderen Coronaviren gelten, wurden mit Wildtyp und den Reportergen-exprimierenden FCoV in vitro infiziert. Die Infektion von monozytären Zellen konnte ausschließlich mit Hilfe der GFP exprimierenden Mutanten nachgewiesen werden.In this study a reverse genetic system for a serotype I feline coronavirus was established. The following results were obtained: 1) As basis for the establishment of a reverse genetic system the complete genomic sequence of FCoV strain Black was determined. The sequence analysis revealed the typical genome organization for coronavirus. 2) For the generation of an infectious clone the full-length FCoV strain Black cDNA was introduced into vaccinia virus as cloning vector. The FCoV cDNA was used for in vitro transcription and the synthesized RNA electroporated into cells. Recombinant FCoV could be detected which showed that the in vitro transcribed RNA was infectious and could initiate the coronavirus replication cycle. The resulting recombinant virus was analyzed in feline cell culture using different approaches. It could be demonstrated that the properties of the recombinant virus are indistinguishable from those of FCoV wild type virus strain Black. 3) In subsequent experiments based on the newly established reverse genetic system, two reporter gene expressing recombinant FCoV were generated. The accessory genes 3abc were replaced by genes encoding the green fluorescent protein (GFP) and Renilla luciferase. The stable expression of these reporter genes was demonstrated in feline cell culture. 4) Feline monocytes, macrophages and dendritic cells (DCs) which are considered to be important target cells for FCoV and other coronaviruses were infected with the wildtype and the reporter gene expressing FCoV. The infection of the monocytic cells was exclusively shown with the GFP expressing mutant

Clostridium difficile Infections amongst Patients with Haematological Malignancies: A Data Linkage Study

OBJECTIVES: Identify risk factors for Clostridium difficile infection (CDI) and assess CDI outcomes among Australian patients with a haematological malignancy. METHODS: A retrospective cohort study involving all patients admitted to hospitals in Western Australia with a haematological malignancy from July 2011 to June 2012. Hospital admission data were linked with all hospital investigated CDI case data. Potential risk factors were assessed by logistic regression. The risk of death within 60 and 90 days of CDI was assessed by Cox Proportional Hazards regression. RESULTS: There were 2085 patients of whom 65 had at least one CDI. Twenty percent of CDI cases were either community-acquired, indeterminate source or had only single-day admissions in the 28 days prior to CDI. Using logistic regression, having acute lymphocytic leukaemia, neutropenia and having had bacterial pneumonia or another bacterial infection were associated with CDI. CDI was associated with an increased risk of death within 60 and 90 days post CDI, but only two deaths had CDI recorded as an antecedent factor. Ribotyping information was available for 33 of the 65 CDIs. There were 19 different ribotypes identified. CONCLUSIONS: Neutropenia was strongly associated with CDI. While having CDI is a risk factor for death, in many cases it may not be a direct contributor to death but may reflect patients having higher morbidity. A wide variety of C. difficile ribotypes were found and community-acquired infection may be under-estimated in these patients

Hospital-acquired infections in the COVID-19 pandemic: a systematic scoping review protocol

Amidst the ongoing COVID19 pandemic, infection prevention and control (IPC) guidelines aimed at preventing nosocomial transmission of the novel Sars-CoV-2 virus have involved even more rigorous interventions such as the screening of healthcare workers and visitors, universal masking policies and patient segregation, typically in addition to the previously described IPC strategies for hospital-acquired infections (HAI). It is unknown how these additional measures have influenced rates of HAIs during the Covid-19 pandemic. The aim of this systematic scoping review is to delineate the range and any changes in the incidence and prevalence of HAIs during the COVID-19 pandemic

Social and community networks influence dietary attitudes in regional New South Wales, Australia

INTRODUCTION Rural populations in Australia have a higher prevalence of obesity, cardiovascular disease, type II diabetes and some cancers. The purpose of the present study was to determine associations between socioeconomic characteristics (socioeconomic position, income, wealth, debt, occupation, social network diversity), dietary attitudes, and fruit and vegetable intake for people living rurally in Australia. METHOD A community based cross-sectional survey between February and July 2018 of 326 adults (median age 57 years, range 20-90 years, 64.4% female) who attended rural shows in four rural towns in south-eastern New South Wales, supplemented with data from patients attending general practices in two additional towns. Participants completed a questionnaire that recorded self-reported daily consumption of fruit and vegetables, a dietary attitude score, and items measuring social and economic circumstances. RESULTS Using multivariable regression analysis, the odds of meeting Australian fruit intake guidelines was 13% higher for each unit increase in dietary attitude score (odds ratio (OR)=1.13, 95% confidence interval (CI)=1.03-1.23). The odds of meeting vegetable intake guidelines were 19% higher for each unit increase in score (OR=1.19, 95%CI=1.09-1.31). Social and economic factors were not independently associated with fruit or vegetable intake. Dietary attitude score, in turn, increased on average by 0.07 points (95%CI=0.01-0.12) for each additional occupation type among the participants' social networks. For women who socialised regularly in small towns the score was 1.97 points higher (95%CI=0.93-3.00). Men in outer regional areas were more likely to meet vegetable intake guidelines than men in inner regional areas, whereas women in outer regional areas were more likely to meet fruit intake guidelines than women in inner regional areas. CONCLUSIONS Greater fruit and vegetable intake was predicted by healthier dietary attitudes which in turn were related to social and community connections, rather than economic factors

Most common diagnoses associated with perinatal admissions 29 days-1year.

<p>Most common diagnoses associated with perinatal admissions 29 days-1year.</p

Most common diagnoses associated with perinatal admissions 0–28 days.

<p>Most common diagnoses associated with perinatal admissions 0–28 days.</p