Next-generation sequencing of cervical DNA detects human papillomavirus types not detected by commercial kits

Background Human papillomavirus (HPV) is the aetiological agent for cervical cancer and genital warts. Concurrent HPV and HIV infection in the South African population is high. HIV positive (+) women are often infected with multiple, rare and undetermined HPV types. Data on HPV incidence and genotype distribution are based on commercial HPV detection kits, but these kits may not detect all HPV types in HIV + women. The objectives of this study were to (i) identify the HPV types not detected by commercial genotyping kits present in a cervical specimen from an HIV positive South African woman using next generation sequencing, and (ii) determine if these types were prevalent in a cohort of HIV-infected South African women. Methods Total DNA was isolated from 109 cervical specimens from South African HIV + women. A specimen within this cohort representing a complex multiple HPV infection, with 12 HPV genotypes detected by the Roche Linear Array HPV genotyping (LA) kit, was selected for next generation sequencing analysis. All HPV types present in this cervical specimen were identified by Illumina sequencing of the extracted DNA following rolling circle amplification. The prevalence of the HPV types identified by sequencing, but not included in the Roche LA, was then determined in the 109 HIV positive South African women by type-specific PCR. Results Illumina sequencing identified a total of 16 HPV genotypes in the selected specimen, with four genotypes (HPV-30, 74, 86 and 90) not included in the commercial kit. The prevalence's of HPV-30, 74, 86 and 90 in 109 HIV positive South African women were found to be 14.6 %, 12.8 %, 4.6 % and 8.3 % respectively. Conclusions Our results indicate that there are HPV types, with substantial prevalence, in HIV positive women not being detected in molecular epidemiology studies using commercial kits. The significance of these types in relation to cervical disease remains to be investigated.

Stability studies of HIV-1 Pr55gag virus-like particles made in insect cells after storage in various formulation media

Background: HIV-1 Pr55gag virus-like particles (VLPs) expressed by baculovirus in insect cells are considered to be a very promising HIV-1 vaccine candidate, as they have been shown to elicit broad cellular immune responses when tested in animals, particularly when used as a boost to DNA or BCG vaccines. However, it is important for the VLPs to retain their structure for them to be fully functional and effective. The medium in which the VLPs are formulated and the temperature at which they are stored are two important factors affecting their stability. FINDINGS We describe the screening of 3 different readily available formulation media (sorbitol, sucrose and trehalose) for their ability to stabilise HIV-1 Pr55gag VLPs during prolonged storage. Transmission electron microscopy (TEM) was done on VLPs stored at two different concentrations of the media at three different temperatures (4[degree sign]C, --20[degree sign]C and -70[degree sign]C) over different time periods, and the appearance of the VLPs was compared. VLPs stored in 15% trehalose at -70[degree sign]C retained their original appearance the most effectively over a period of 12 months. VLPs stored in 5% trehalose, sorbitol or sucrose were not all intact even after 1 month storage at the temperatures tested. In addition, we showed that VLPs stored under these conditions were able to be frozen and re-thawed twice before showing changes in their appearance. Conclusions Although the inclusion of other analytical tools are essential to validate these preliminary findings, storage in 15% trehalose at -70[degree sign]C for 12 months is most effective in retaining VLP stability.

Setting up a platform for plant-based influenza virus vaccine production in South Africa

Background During a global influenza pandemic, the vaccine requirements of developing countries can surpass their supply capabilities, if these exist at all, compelling them to rely on developed countries for stocks that may not be available in time. There is thus a need for developing countries in general to produce their own pandemic and possibly seasonal influenza vaccines. Here we describe the development of a plant-based platform for producing influenza vaccines locally, in South Africa. Plant-produced influenza vaccine candidates are quicker to develop and potentially cheaper than egg-produced influenza vaccines, and their production can be rapidly upscaled. In this study, we investigated the feasibility of producing a vaccine to the highly pathogenic avian influenza A subtype H5N1 virus, the most generally virulent influenza virus identified to date. Two variants of the haemagglutinin (HA) surface glycoprotein gene were synthesised for optimum expression in plants: these were the full-length HA gene (H5) and a truncated form lacking the transmembrane domain (H5tr). The genes were cloned into a panel of Agrobacterium tumefaciens binary plant expression vectors in order to test HA accumulation in different cell compartments. The constructs were transiently expressed in tobacco by means of agroinfiltration. Stable transgenic tobacco plants were also generated to provide seed for stable storage of the material as a pre-pandemic strategy. Results For both transient and transgenic expression systems the highest accumulation of full-length H5 protein occurred in the apoplastic spaces, while the highest accumulation of H5tr was in the endoplasmic reticulum. The H5 proteins were produced at relatively high concentrations in both systems. Following partial purification, haemagglutination and haemagglutination inhibition tests indicated that the conformation of the plant-produced HA variants was correct and the proteins were functional. The immunisation of chickens and mice with the candidate vaccines elicited HA-specific antibody responses. Conclusions We managed, after synthesis of two versions of a single gene, to produce by transient and transgenic expression in plants, two variants of a highly pathogenic avian influenza virus HA protein which could have vaccine potential. This is a proof of principle of the potential of plant-produced influenza vaccines as a feasible pandemic response strategy for South Africa and other developing countries.

Recombination hotspots and host susceptibility modulate the adaptive value of recombination during maize streak virus evolution

Background Maize streak virus -strain A (MSV-A; Genus Mastrevirus, Family Geminiviridae), the maize-adapted strain of MSV that causes maize streak disease throughout sub-Saharan Africa, probably arose between 100 and 200 years ago via homologous recombination between two MSV strains adapted to wild grasses. MSV recombination experiments and analyses of natural MSV recombination patterns have revealed that this recombination event entailed the exchange of the movement protein - coat protein gene cassette, bounded by the two genomic regions most prone to recombination in mastrevirus genomes; the first surrounding the virion-strand origin of replication, and the second around the interface between the coat protein gene and the short intergenic region. Therefore, aside from the likely adaptive advantages presented by a modular exchange of this cassette, these specific breakpoints may have been largely predetermined by the underlying mechanisms of mastrevirus recombination. To investigate this hypothesis, we constructed artificial, low-fitness, reciprocal chimaeric MSV genomes using alternating genomic segments from two MSV strains; a grass-adapted MSV-B, and a maize-adapted MSV-A. Between them, each pair of reciprocal chimaeric genomes represented all of the genetic material required to reconstruct - via recombination - the highly maize-adapted MSV-A genotype, MSV-MatA. We then co-infected a selection of differentially MSV-resistant maize genotypes with pairs of reciprocal chimaeras to determine the efficiency with which recombination would give rise to high-fitness progeny genomes resembling MSV-MatA. Results Recombinants resembling MSV-MatA invariably arose in all of our experiments. However, the accuracy and efficiency with which the MSV-MatA genotype was recovered across all replicates of each experiment depended on the MSV susceptibility of the maize genotypes used and the precise positions - in relation to known recombination hotspots - of the breakpoints required to re-create MSV-MatA. Although the MSV-sensitive maize genotype gave rise to the greatest variety of recombinants, the measured fitness of each of these recombinants correlated with their similarity to MSV-MatA. Conclusions The mechanistic predispositions of different MSV genomic regions to recombination can strongly influence the accessibility of high-fitness MSV recombinants. The frequency with which the fittest recombinant MSV genomes arise also correlates directly with the escalating selection pressures imposed by increasingly MSV-resistant maize hosts.

Children's Literature Digital Resource (CLDR)

Children’s Literature Digital Resources incorporates primary texts published from white settlement to 1945, including children’s and young adult fiction, poetry, short stories, and picture books. This collection is supported by selected secondary material. The objective is to provide a centralised access point for information about Australian children's literature and writers and a growing body of full-text primary resources. Four key aims are: * To establish an important digital facility for research, teaching, and information provision around Australian children’s literature; * To provide access to a wide range of high-quality full-text data, both primary and secondary resources; * To provide access to essential library and research information infrastructure and facilities for established and emerging researchers in the fields of Humanities and Education; To enable research while preserving important heritage material. The collection contains texts digitised for AustLit through cooperation with various Australian libraries. The collection includes children’s and young adult fiction, poetry, picture books, short stories, and critical articles relating to relevant primary texts. Authors of primary sources include Irene Cheyne, E. W. Cole, Richard Rowe, Lillian M. Pyke, and Dorothy Wall. Secondary sources include critical works by Clare Bradford, Heather Scutter, Kerry White, Sharyn Pearce, and Marcie Muir. These full-text materials are keyword searchable (both within individual texts and across the CLDR corpus) and can be downloaded for research purposes. As well as digitising primary and secondary material, the project locates and provides pathways to existing online resources or internet publications to enhance AustLit's Children's Literature subset. These resources include both primary and secondary texts.

Asian-Australian Children’s Literature and Publishing (AACLAP)

Led by Queensland University of Technology, the Asian-Australian Children’s Literature and Publishing (AACLAP)research project investigates and records details of Australian children’s literature that is set in Asia and/or that represents Asian-Australian cultures and experiences and literature that is published in selected Asian languages. This includes East Asia, Southeast Asia, South Asia, and the Bay of Bengal. The AACLAP dataset is a comprehensive collection of agent and work records related to ’Asia’, including, but not limited to, autobiography, fiction, criticism, poetry, drama, short stories, and picture books, published during a forty-year period from 1970 to 2010. The dataset provide valuable primary and secondary sources that are important for developing literature-focused educational programs in line with the national government’s push for Asia Literacy. AACLAP is a subset of AustLit, the virtual research environment and information resource for Australian literary, print, and narrative culture scholars, students, and the public.

Open access to judgments : Creative Commons licences and the Australian courts

Internet technologies have fundamentally changed the way we obtain access to legal documents and information about the law. However, for judgments of courts and tribunals, copyright management and licensing practices have not kept pace with the digital and online technologies which are now ubiquitous in the web 2.0 era. Under the provisions of the Copyright Act 1968 and the licensing statements on the Australian courts’ websites, judgments may generally be read online, downloaded, reproduced and printed out for personal, non-commercial use or ”in house” use by an organisation. However, beyond these permitted acts, the extent to which judgments can be copied and distributed in digital form online remains unclear. Open content licences (in particular, the Creative Commons (CC) licences) offer an effective mechanism for managing copyright in judgments in a manner that supports their wide public dissemination and reuse while also protecting their integrity and accuracy.

Signal transduction mechanisms underlying growth hormone receptor action

Our understanding of the mechanisms of action of GH and its receptor, the GHR, has advanced significantly in the last decade and has provided some important surprises. It is now clear that the GH-GHR axis activates a number of inter-related signalling pathways, not all of which are dependent on the intracellular tyrosine kinase, JAK2 as originally postulated. JAK2-independent pathways, mediated via the Src family kinases, together with a number of negative regulators of GH signalling and emerging cross-talk mechanisms with other growth factor receptors, provide a complex array of mechanisms that are capable of fine-tuning responses to GH in a cell context dependent manner. Additionally, it is also now clear that GH and the GHR can translocate to the nucleus of target cells and initiate, as yet not well defined, nuclear responses. Continued emphasis on elucidation of these complex mechanisms is critical to provide further insights into the diverse physiological and pathophysiological effects of GH.

Socio-structural influences on the work participation of refugees : an exploratory systematic mixed studies review

This systematic mixed studies review aimed at synthesizing evidence from studies related to the influences on the work participation of people with refugee status (PWRS). The review focused on the role of proximal socio-structural barriers on work participation by PWRS while foregrounding related distal, intermediate, proximal, and meta-systemic influences. For the systematic search of the literature, we focused on databases that addressed work, well-being, and social policy in refugee populations, including, Medline, CINAHL, PsycInfo, Web of Science, Scopus, and Sociological Abstracts. Of the studies reviewed, 16 of 39 met the inclusion criteria and were retained for the final analysis. We performed a narrative synthesis of the evidence on barriers to work participation by PWRS, interlinking clusters of barriers potent to their effects on work participation. Findings from the narrative synthesis suggest that proximal factors, those at point of entry to the labor market, influence work participation more directly than distal or intermediate factors. Distal and intermediate factors achieve their effects on work participation by PWRS primarily through meta-systemic interlinkages, including host-country documentation and refugee administration provisions.

Study protocol : Screening and treatment of alcohol-related trauma (START): a randomised controlled trial

Background: The incidence of mandibular fractures in the Northern Territory of Australia is very high, especially among Indigenous people. Alcohol intoxication is implicated in the majority of facial injuries, and substance use is therefore an important target for secondary prevention. The current study tests the efficacy of a brief therapy, Motivational Care Planning, in improving wellbeing and substance misuse in youth and adults hospitalised with alcohol-related facial trauma. Methods and design: The study is a randomised controlled trial with 6 months of follow-up, to examine the effectiveness of a brief and culturally adapted intervention in improving outcomes for trauma patients with at-risk drinking admitted to the Royal Darwin Hospital maxillofacial surgery unit. Potential participants are identified using AUDIT-C questionnaire. Eligible participants are randomised to either Motivational Care Planning (MCP) or Treatment as Usual (TAU). The outcome measures will include quantity and frequency of alcohol and other substance use by Timeline Followback. The recruitment target is 154 participants, which with 20% dropout, is hoped to provide 124 people receiving treatment and follow-up. Discussion: This project introduces screening and brief interventions for high-risk drinkers admitted to the hospital with facial trauma. It introduces a practical approach to integrating brief interventions in the hospital setting, and has potential to demonstrate significant benefits for at-risk drinkers with facial trauma.

Recruiting and engaging new mothers in nutrition research studies : lessons from the Australian NOURISH randomised controlled trial

Background: Despite important implications for the budgets, statistical power and generalisability of research findings, detailed reports of recruitment and retention in randomised controlled trials (RCTs) are rare. The NOURISH RCT evaluated a community-based intervention for first-time mothers that promoted protective infant feeding practices as a primary prevention strategy for childhood obesity. The aim of this paper is to provide a detailed description and evaluation of the recruitment and retention strategies used. Methods: A two stage recruitment process designed to provide a consecutive sampling framework was used. First time mothers delivering healthy term infants were initially approached in postnatal wards of the major maternity services in two Australian cities for consent to later contact (Stage 1). When infants were about four months old mothers were re-contacted by mail for enrolment (Stage 2), baseline measurements (Time 1) and subsequent random allocation to the intervention or control condition. Outcomes were assessed at infant ages 14 months (Time 2) and 24 months (Time 3). Results: At Stage 1, 86% of eligible mothers were approached and of these women, 76% consented to later contact. At Stage 2, 3% had become ineligible and 76% could be recontacted. Of the latter, 44% consented to full enrolment and were allocated. This represented 21% of mothers screened as eligible at Stage 1. Retention at Time 3 was 78%. Mothers who did not consent or discontinued the study were younger and less likely to have a university education. Conclusions: The consent and retention rates of our sample of first time mothers are comparable with or better than other similar studies. The recruitment strategy used allowed for detailed information from non-consenters to be collected; thus selection bias could be estimated. Recommendations for future studies include being able to contact participants via mobile phone (particular text messaging), offering home visits to reduce participant burden and considering the use of financial incentives to support participant retention.

What makes an LMS effective : a synthesis of current literature

There is a growing number of organizations and universities now utilising e-learning practices in their teaching and learning programs. These systems have allowed for knowledge sharing and provide opportunities for users to have access to learning materials regardless of time and place. However, while the uptake of these systems is quite high, there is little research into the effectiveness of such systems, particularly in higher education. This paper investigates the methods that are used to study the effectiveness of e-learning systems and the factors that are critical for the success of a learning management system (LMS). Five major success categories are identified in this study and explained in depth. These are the teacher, student, LMS design, learning materials and external support.

Spatial clusters of suicide in Australia

Background: Understanding the spatial distribution of suicide can inform the planning, implementation and evaluation of suicide prevention activity. This study explored spatial clusters of suicide in Australia, and investigated likely socio-demographic determinants of these clusters. Methods: National suicide and population data at a statistical local area (SLA) level were obtained from the Australian Bureau of Statistics for the period of 1999 to 2003. Standardised mortality ratios (SMR) were calculated at the SLA level, and Geographic Information System (GIS) techniques were applied to investigate the geographical distribution of suicides and detect clusters of high risk in Australia. Results: Male suicide incidence was relatively high in the northeast of Australia, and parts of the east coast, central and southeast inland, compared with the national average. Among the total male population and males aged 15 to 34, Mornington Shire had the whole or a part of primary high risk cluster for suicide, followed by the Bathurst-Melville area, one of the secondary clusters in the north coastal area of the Northern Territory. Other secondary clusters changed with the selection of cluster radius and age group. For males aged 35 to 54 years, only one cluster in the east of the country was identified. There was only one significant female suicide cluster near Melbourne while other SLAs had very few female suicide cases and were not identified as clusters. Male suicide clusters had a higher proportion of Indigenous population and lower median socio-economic index for area (SEIFA) than the national average, but their shapes changed with selection of maximum cluster radii setting. Conclusion: This study found high suicide risk clusters at the SLA level in Australia, which appeared to be associated with lower median socio-economic status and higher proportion of Indigenous population. Future suicide prevention programs should focus on these high risk areas.

Iterative development of MobileMums : a physical activity intervention for women with young children.

Background To describe the iterative development process and final version of ‘MobileMums’: a physical activity intervention for women with young children (<5 years) delivered primarily via mobile telephone (mHealth) short messaging service (SMS). Methods MobileMums development followed the five steps outlined in the mHealth development and evaluation framework: 1) conceptualization (critique of literature and theory); 2) formative research (focus groups, n= 48); 3) pre-testing (qualitative pilot of intervention components, n= 12); 4) pilot testing (pilot RCT, n= 88); and, 5) qualitative evaluation of the refined intervention (n= 6). Results Key findings identified throughout the development process that shaped the MobileMums program were the need for: behaviour change techniques to be grounded in Social Cognitive Theory; tailored SMS content; two-way SMS interaction; rapport between SMS sender and recipient; an automated software platform to generate and send SMS; and, flexibility in location of a face-to-face delivered component. Conclusions The final version of MobileMums is flexible and adaptive to individual participant’s physical activity goals, expectations and environment. MobileMums is being evaluated in a community-based randomised controlled efficacy trial (ACTRN12611000481976).