Characterisation Of Adeno-Associated Virus DNA contaminants and Development of a P5 promoter replacement, high purity Adeno-Associated Virus production system

Gene therapy is the transfer of nucleic acids for therapeutic benefit, a process potentially achieved through several delivery methods. One of the most common gene therapy delivery modalities is adeno associated virus (AAV). The availability of a system to produce large quantities of AAV, and of diverse AAV serotypes, that can infect different cell types makes AAV very attractive for widespread clinical implementation. Viruses like AAV that are manipulated to become highly useful delivery tools rely on complex biological systems for production. It is imperative that these systems are well characterised and designed to be highly safe and efficacious. This PhD thesis investigated nucleic acid contamination of AAV preparations, looking at the production system as a source, and the problems that this may pose for AAV infected cells. Specific regions of DNA from AAV producer plasmids were incorporated into virions more abundantly. These were found adjacent to the outside of the inverted terminal repeats (ITRs) that flank the expression cassette, and on the plasmid containing the replication and capsid genes, directly upstream of the P5 promoter which drives expression of the replication proteins, REP78 and REP68. Contaminant sequences were found to be transferred into transduced cells during AAV infection, persistent within cells and transcriptionally active. Investigation of P5 upstream contaminants showed potential for protein to be produced persistently from contaminant sequences both in vitro and in vivo. Finally, precise sequence regions causing DNA incorporation upstream of the P5 promoter were examined. This information was used to produce alternative production 3 reagents that, in their first iteration, provided proof of principle of P5-related contaminant removal from the final AAV product, and in additional iterations resulted in a modified P5 promoter (P5-HS). This promoter limited upstream contamination thereby resulting in higher purity AAV, whilst retaining vector yields equivalent to the original AAV production system

Is Financial Capability Related to the Effective Use of Debt in Australia?

Australians’ high use of personal debt is, in part, attributable to the relaxation of the financial services regulation. There is concern that while debt has the potential to increase a person’s wealth, if used ineffectively it can have the opposite effect. This paper details a study of 680 Australians to ascertain whether their financial capability is related to the effective use of personal debt. The findings suggest that it appears people with greater financial capability are more likely to use debt effectively

The iPad cohort of 2013: A multiple stakeholder view of the planning, design and implementation of a first year curriculum integrating tablets

Abstract In a quest to develop strategies to improve teaching and learning outcomes and student engagement across the business school, a pilot program was devised that involved the provision of tablet computers (iPads

A Scalable System for Production of Functional Pancreatic Progenitors from Human Embryonic Stem Cells

Development of a human embryonic stem cell (hESC)-based therapy for type 1 diabetes will require the translation of proof-of-principle concepts into a scalable, controlled, and regulated cell manufacturing process. We have previously demonstrated that hESC can be directed to differentiate into pancreatic progenitors that mature into functional glucose-responsive, insulin-secreting cells in vivo. In this study we describe hESC expansion and banking methods and a suspension-based differentiation system, which together underpin an integrated scalable manufacturing process for producing pancreatic progenitors. This system has been optimized for the CyT49 cell line. Accordingly, qualified large-scale single-cell master and working cGMP cell banks of CyT49 have been generated to provide a virtually unlimited starting resource for manufacturing. Upon thaw from these banks, we expanded CyT49 for two weeks in an adherent culture format that achieves 50–100 fold expansion per week. Undifferentiated CyT49 were then aggregated into clusters in dynamic rotational suspension culture, followed by differentiation en masse for two weeks with a four-stage protocol. Numerous scaled differentiation runs generated reproducible and defined population compositions highly enriched for pancreatic cell lineages, as shown by examining mRNA expression at each stage of differentiation and flow cytometry of the final population. Islet-like tissue containing glucose-responsive, insulin-secreting cells was generated upon implantation into mice. By four- to five-months post-engraftment, mature neo-pancreatic tissue was sufficient to protect against streptozotocin (STZ)-induced hyperglycemia. In summary, we have developed a tractable manufacturing process for the generation of functional pancreatic progenitors from hESC on a scale amenable to clinical entry

Editorial

The financial planning industry is at a critical phase of its development. The industry is still in its infancy and yet the impact of financial advice is far reaching in our society. After many years of rapid, yet dislocated, growth and various controversies, the industry is embarking on a fascinating journey towards becoming a profession. Recently the move to a fee for service basis of operation, mandatory professional indemnity insurance, formal codes of conduct and higher barriers to entry in regards to educational and professional experience requirements are key signs of development of a financial planning profession for the future

Financial Planning and Financial Instruments: 2013 in Review, 2014 in Prospect

The Global Financial Crisis (GFC); changes in regulation; issues in public perception and trust have contributed to a troubled 2013 for financial planners. As financial planning and wealth management providers seek to bolster their professional status, the Financial Planning Education Council’s National Curriculum and Accreditation Framework and ASIC’s minimum training requirements are also a space to watch. In prospect, 2014 will offer opportunities and challenges in the form of a changed government; regulatory changes; accreditation challenges and the hope of the recruitment of strong financial planning and wealth management professionals for the future

Editorial: Special Issue on Financial Planning

The special edition presents current research in the area of financial planning. With the continual upheaval in global financial markets (including Australia), the general trend towards self-funded retirement and less reliance on the state, financial crises and the continual regulatory changes in the financial markets, this issue is timely and topical. This is the third special edition on financial planning, providing an excellent outlet for quality research in financial planning

Factors that Influence Financial Capability and Effectiveness: Exploring Financial Counsellors’ Perspectives

The financial capability of the community has become a topical issue in recent years around the globe. The role financial education plays improving financial capability is recognised by the G20 countries. With a plethora of programs, workshops, seminars and other resources available in the community, we find that there is little research that measures the impact/usefulness of these programs including what should be included in such programs. This study seeks to contribute to the understanding of the underlying determinants of financial capability by exploring the factors that influence this. Focus groups were conducted with financial counsellors in Queensland, Australia to explore factors that inhibit and promote financial capability. This included factors that impede the development of capability and the others that impact on the development of intention to engage in financially effective behaviour. We report that financial counsellors view confidence, self-esteem and self-belief as equally important determinants of financial capability. Also, gender and family socio economic status influence an individual’s ability to engage in financially effective behaviour. The results also found that adopting a short-term focus, rather than future orientation, is a key inhibitor of financial effectiveness. Consequently it is suggested that those developing financial capability programs address these behavioural and contextual factors rather than concentrating purely on literacy