High level protein expression in plants through the use of a novel autonomously replicating geminivirus shuttle vector

We constructed a novel autonomously replicating gene expression shuttle vector, with the aim of developing a system for transiently expressing proteins at levels useful for commercial production of vaccines and other proteins in plants. The vector, pRIC, is based on the mild strain of the geminivirus Bean yellow dwarf virus (BeYDV-m) and is replicationally released into plant cells from a recombinant Agrobacterium tumefaciens Ti plasmid. pRIC differs from most other geminivirus-based vectors in that the BeYDV replication-associated elements were included in cis rather than from a co-transfected plasmid, while the BeYDV capsid protein (CP) and movement protein (MP) genes were replaced by an antigen encoding transgene expression cassette derived from the non-replicating A. tumefaciens vector, pTRAc. We tested vector efficacy in Nicotiana benthamiana by comparing transient cytoplasmic expression between pRIC and pTRAc constructs encoding either enhanced green fluorescent protein (EGFP) or the subunit vaccine antigens, human papillomavirus subtype 16 (HPV-16) major CP L1 and human immunodeficiency virus subtype C p24 antigen. The pRIC constructs were amplified in planta by up to two orders of magnitude by replication, while 50% more HPV-16 L1 and three- to seven-fold more EGFP and HIV-1 p24 were expressed from pRIC than from pTRAc. Vector replication was shown to be correlated with increased protein expression. We anticipate that this new high-yielding plant expression vector will contribute towards the development of a viable plant production platform for vaccine candidates and other pharmaceuticals. © 2009 Blackwell Publishing Ltd.

Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production

Background Insect baculovirus-produced Human immunodeficiency virus type 1 (HIV-1) Gag virus-like-particles (VLPs) stimulate good humoral and cell-mediated immune responses in animals and are thought to be suitable as a vaccine candidate. Drawbacks to this production system include contamination of VLP preparations with baculovirus and the necessity for routine maintenance of infectious baculovirus stock. We used piggyBac transposition as a novel method to create transgenic insect cell lines for continuous VLP production as an alternative to the baculovirus system. Results Transgenic cell lines maintained stable gag transgene integration and expression up to 100 cell passages, and although the level of VLPs produced was low compared to baculovirus-produced VLPs, they appeared similar in size and morphology to baculovirus-expressed VLPs. In a murine immunogenicity study, whereas baculovirus-produced VLPs elicited good CD4 immune responses in mice when used to boost a prime with a DNA vaccine, no boost response was elicited by transgenically produced VLPs. Conclusion Transgenic insect cells are stable and can produce HIV Pr55 Gag VLPs for over 100 passages: this novel result may simplify strategies aimed at making protein subunit vaccines for HIV. Immunogenicity of the Gag VLPs in mice was less than that of baculovirus-produced VLPs, which may be due to lack of baculovirus glycoprotein incorporation in the transgenic cell VLPs. Improved yield and immunogenicity of transgenic cell-produced VLPs may be achieved with the addition of further genetic elements into the piggyBac integron.

An investigation into the use of human papillomavirus type 16 virus-like particles as a delivery vector system for foreign proteins: N- and C-terminal fusion of GFP to the L1 and L2 capsid proteins

Development of vaccine strategies against human papillomavirus (HPV), which causes cervical cancer, is a priority. We investigated the use of virus-like particles (VLPs) of the most prevalent type, HPV-16, as carriers of foreign proteins. Green fluorescent protein (GFP) was fused to the N or C terminus of both L1 and L2, with L2 chimeras being co-expressed with native L1. Purified chimaeric VLPs were comparable in size (∼55 nm) to native HPV VLPs. Conformation-specific monoclonal antibodies (Mabs) bound to the VLPs, thereby indicating that they possibly retain their antigenicity. In addition, all of the VLPs encapsidated DNA in the range of 6-8 kb. © 2007 Springer-Verlag.

Community-based asylum seekers’ use of primary health care services in Melbourne

Objective: To investigate primary health care service utilisation and health presentations among asylum seekers living in Melbourne. Design and setting: Retrospective audit of files of people who attended three Melbourne asylum-seeker health clinics between 1 July 2005 and 30 June 2006. Main outcome measures: Rates of reasons for the encounter, diagnostic tests or investigations required, treatments prescribed and referrals. Results: Data were collected from 998 consultations corresponding to 341 people. Eighty-eight per cent of visits involved people with no Medicare access, owing to their visa status. The most common reasons for the encounter were general and unspecified symptoms or problems (rate, 59.9 per 100 encounters; 95% CI, 55–65), followed by musculoskeletal conditions (27.1; 95% CI, 24–30), and psychological problems (26.5; 95% CI, 23–30). The rate of referrals was 18.3 per 100 encounters (95% CI, 16–21). Conclusions: The three clinics providing services to asylum seekers in Melbourne are delivering care to a considerable number of people with complex health needs. A substantial number of asylum seekers present to clinics with psychological and social problems. Most cannot access government-subsidised health care. This must be addressed urgently by policy change at the federal and state and territory levels.

Rating the Audience : the Business of Media

Knowing, measuring and understanding media audiences have become a multi-billion dollar business. But the convention that underpins that business, audience ratings, is in crisis. Rating the Audience is the first book to show why and how audience ratings research became a convention, an agreement, and the first to interrogate the ways that agreement is now under threat. Taking a historical approach, the book looks at the evolution of audience ratings and the survey industry. It goes on to analyse today's media environment, looking at the role of the internet and the increased difficulties it presents for measuring audiences. The book covers all the major players and controversies, such as Facebook's privacy rulings and Google's alliance with Nielsen. Offering the first real comparative study, it will be critical for media students and professionals.

ACE research vignette 021 : does the use of “bricolage” make start-ups more innovative?

This series of research vignettes is aimed at sharing current and interesting research findings from our team of international Entrepreneurship researchers. In this vignette, Professor Per Davidsson and Associate Professor Paul Steffens consider the links between entrepreneurial “bricolage” and innovation.

Is there a place for pixels in our practice? Dietitians' attitudes towards the use of photographic dietary records within the nutrition care process

Photographic records of dietary intake (PhDRs) are an innovative method for the dietary assessment and may alleviate the burden of recording intake compared to traditional methods of recording intake. While the performance of PhDRs has been evaluated, no investigation into the application of this method had occurre within dietetic practice. This study examined the attitudes of dietitians towards the use of PhDRs in the provision of nutrition care. A web-based survey on the practices and beliefs with regards to technology use among Dietitians Association of Australia members was conducted in August 2011. Of the 87 dietitians who responded, 86% assessed the intakes of clients as part of individualised medical nutrition therapy, with the diet history the most common method used. The majority (91%) of dietitians surveyed believed that a PhDR would be of use in their current practice to estimate intake. Information contained in the PhDR would primarily be used to obtain a qualitative evaluation of diet (84%) or to supplement an existing assessment method (69%), as opposed to deriving an absolute measure of nutrient intake (31%). Most (87%) indicated that a PhDR would also be beneficial in both the delivery of the intervention and to evaluate and monitor goals and outcomes, while only 46% felt that a PhDR would assist in determining the nutrition diagnosis. This survey highlights the potential for the use of PhDRs within practice. Future endeavours lie in establishing resources which support the inclusion of PhDRs within the nutrition care process.

Adaption and use of a compressible flow code for turbomachinery design

The present paper presents and discusses the use of dierent codes regarding the numerical simulation of a radial-in ow turbine. A radial-in ow turbine test case was selected from published literature [1] and commercial codes (Fluent and CFX) were used to perform the steady-state numerical simulations. An in-house compressible- ow simulation code, Eilmer3 [2] was also adapted in order to make it suitable to perform turbomachinery simulations and preliminary results are presented and discussed. The code itself as well as its adaptation, comprising the addition of terms for the rotating frame of reference, programmable boundary conditions for periodic boundaries and a mixing plane interface between the rotating and non-rotating blocks are also discussed. Several cases with dierent orders of complexity in terms of geometry were considered and the results were compared across the dierent codes. The agreement between these results and published data is also discussed.

Use of focus group technique to investigate infection control practice

Use of focus groups as a technique of inquiry is gaining attention in the area of health-care research. This paper will report on the technique of focus group interviewing to investigate the role of the infection control practitioner. Infection control is examined as a specialty area of health-care practice that has received little research attention to date. Additionally, it is an area of practice that is expanding in response to social, economic and microbiological forces. The focus group technique in this study helped a group of infection control practitioners from urban, regional and rural areas throughout Queensland identify and categorise their daily work activities. The outcomes of this process were then analysed to identify the growth in breadth and complexity of the role of the infection control practitioner in the contemporary health-care environment. Findings indicate that the role of the infection control practitioner in Australia has undergone changes consistent with and reflecting changing models of health-care delivery.

Use of condition-specific patient-reported outcome measures in clinical trials among patients with wrist osteoarthritis : a systematic review

Background. This paper aimed to identify condition-specific patient-reported outcome measures used in clinical trials among people with wrist osteoarthritis and summarise empirical peer-reviewed evidence supporting their reliability, validity, and responsiveness to change. Methods. A systematic review of randomised controlled trials among people with wrist osteoarthritis was undertaken. Studies reporting reliability, validity, or responsiveness were identified using a systematic reverse citation trail audit procedure. Psychometric properties of the instruments were examined against predefined criteria and summarised. Results. Thirteen clinical trials met inclusion criteria. The most common patient-reported outcome was the disabilities of the arm, shoulder, and hand questionnaire (DASH). The DASH, the Michigan Hand Outcomes Questionnaire (MHQ), the Patient Evaluation Measure (PEM), and the Patient-Reported Wrist Evaluation (PRWE) had evidence supporting their reliability, validity, and responsiveness. A post-hoc review of excluded studies revealed the AUSCAN Osteoarthritis Hand Index as another suitable instrument that had favourable reliability, validity, and responsiveness. Conclusions. The DASH, MHQ, and AUSCAN Osteoarthritis Hand Index instruments were supported by the most favourable empirical evidence for validity, reliability, and responsiveness. The PEM and PRWE also had favourable empirical evidence reported for these elements. Further psychometric testing of these instruments among people with wrist osteoarthritis is warranted.

Use of a clay modeling task to reduce chocolate craving

Elaborated Intrusion theory (EI theory; Kavanagh, Andrade, & May, 2005) posits two main cognitive components in craving: associative processes that lead to intrusive thoughts about the craved substance or activity, and elaborative processes supporting mental imagery of the substance or activity. We used a novel visuospatial task to test the hypothesis that visual imagery plays a key role in craving. Experiment 1 showed that spending 10 min constructing shapes from modeling clay (plasticine) reduced participants' craving for chocolate compared with spending 10 min 'letting your mind wander'. Increasing the load on verbal working memory using a mental arithmetic task (counting backwards by threes) did not reduce craving further. Experiment 2 compared effects on craving of a simpler verbal task (counting by ones) and clay modeling. Clay modeling reduced overall craving strength and strength of craving imagery, and reduced the frequency of thoughts about chocolate. The results are consistent with EI theory, showing that craving is reduced by loading the visuospatial sketchpad of working memory but not by loading the phonological loop. Clay modeling might be a useful self-help tool to help manage craving for chocolate, snacks and other foods.

Reproductive health : findings from the Australian Longitudinal Study on Women’s Health. Chapter 3: Use of contraception

Chapter 3: Use of contraception. p15-32 Key findings: This section examines trends in the use of contraception between 1996 and 2006 by women who participated in the surveys for the 1973-1978 cohort of the Australian Longitudinal Study on Women’s Health. 1. The oral contraceptive pill was the most commonly used method of contraception at each survey, but its use decreased over time. 2.Of women who consistently used contraception, 40% used the oral contraceptive pill as their only method of contraception in at least three out of four surveys. 3. The proportion of women using condoms as their only method of contraception remained steady over time (15-18%) but only 3% of all women used condoms only at every survey. 4. The proportion of women using both condoms and the oral contraceptive pill remained steady at 13-14% of all women from Survey 1 to 3, but decreased to 8% of all women at Survey 4. 5. The use of methods other than the oral contraceptive pill and/or condoms increased at Survey 4. 6. The proportion of women using an implant (e.g. Implanon) remained steady between Surveys 3 and 4, with 3% of women using an implant only. Around one third of implant users at Survey 3 continued to use this method at Survey 4. 7. The main reasons for not using contraception at Surveys 3 and 4 were pregnancy, trying to conceive, or no male sexual partners. 8. Women who used contraception were more likely to be in de facto relationships or single, be up to date with Pap tests and have had two or more births. 9. Women who did not use contraception were more likely to be non-drinkers and/or do low levels of exercise, have had one birth and have experienced miscarriage. 10. Contraception changed in expected ways according to reproductive events: women who reported only miscarriages between surveys also stopped using contraception in the same period; most women who did not report reproductive events continued to use the same method of contraception; and women who had a termination tended to switch methods.