Syrian hamster prion protein (PrPC) and a truncated Syrian hamster prion protein lacking the glycosylphosphatidylinositol (GPI) anchor C-terminal signal sequence (GPI-) were expressed in Chinese hamster ovary cells using a glutamine synthetase selection and amplification system. The CHO cell clones expressing the GPI- PrP secreted the majority of the protein into the media, whereas most of the PrP produced by clones expressing the full-length protein with the GPI anchor was located on the cell surface, as demonstrated by its release upon treatment with phosphatidylinositol-specific phospholipase C (PIPLC). A cell clone that expressed the highest levels of full length PrP was subcloned to obtain clone 30C3-1. PrP from clone 30C3-1 was shown to be sensitive to proteolysis by proteinase K and to react with monoclonal and polyclonal antibodies that recognize native PrPC. The recombinant PrP migrated as a diffuse band of 19-40 kDa but removal of the N-linked oligosaccharides with peptide N-glycosidase F (PNGase F) revealed three protein species of 19, 17 and 15 kDa. The 19 kDa band corresponding to deglycosylated full-length PrP was quantified and found to be expressed at a level approximately 14-fold higher than that of PrPC found in Syrian hamster brain.

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Background Antibiotic resistance is a major global threat to mankind. Inappropriate antibiotic use has been identified as one of the greatest contributors to antibiotic resistance. There is currently a paucity of data on antibiotic use in aged care facilities, globally and in the Australian setting. Aim To investigate the current antibiotic prescribing practices and use in aged care facilities in the Riverina, New South Wales (NSW), and metropolitan Australian Capital Territory (ACT). Method Care managers of aged care facilities in the ACT and Riverina, as well as pharmacists who service aged care facilities in regional NSW and ACT, were asked to complete a questionnaire relating to antibiotic use in aged care facilities. Data obtained were subjected to statistical analysis. Results Most facilities in the ACT (85%) and NSW (90%) had some form of protocol in place for the prescribing and obtaining of antibiotics for residents. The most commonly prescribed antibiotic across participating facilities was cephalexin and the most common indication for antibiotic use was urinary tract infection. No statistically significant differences in antibiotic use and antibiotic prescribing practices were observed between metropolitan ACT and regional NSW aged care facilities. There was a statistically significant association between facilities with residents taking antibiotics for ongoing conditions and those that have experienced situations in the past 12 months in which the initially prescribed antibiotic did not work and another had to be used (p = 0.004944). Conclusion The data from this study can inform further studies that may be used to inform the development of antimicrobial stewardship initiatives in aged care facilities.