Construct design for efficient, effective and high-throughput gene silencing in plants

Post-transcriptional silencing of plant genes using anti-sense or co-suppression constructs usually results in only a modest proportion of silenced individuals. Recent work has demonstrated the potential for constructs encoding self-complementary 'hairpin' RNA (hpRNA) to efficiently silence genes. In this study we examine design rules for efficient gene silencing, in terms of both the proportion of independent transgenic plants showing silencing, and the degree of silencing. Using hpRNA constructs containing sense/anti-sense arms ranging from 98 to 853 nt gave efficient silencing in a wide range of plant species, and inclusion of an intron in these constructs had a consistently enhancing effect. Intron-containing constructs (ihpRNA) generally gave 90-100% of independent transgenic plants showing silencing. The degree of silencing with these constructs was much greater than that obtained using either co-suppression or anti-sense constructs. We have made a generic vector, pHANNIBAL, that allows a simple, single PCR product from a gene of interest to be easily converted into a highly effective ihpRNA silencing construct. We have also created a high-throughput vector, pHELLSGATE, that should facilitate the cloning of gene libraries or large numbers of defined genes, such as those in EST collections, using an in vitro recombinase system. This system may facilitate the large-scale determination and discovery of plant gene functions in the same way as RNAi is being used to examine gene function in Caenorhabditis elegans.

Carrot red leaf and carrot mottle viruses : observations on the composition of the particles in single and mixed infections

Particles of carrot red leaf virus (CRLV; luteovirus group) purified from chervil (Anthriscus cerefolium) contain a single ssRNA species of mol. wt. about 1.8 x 106 and a major protein of mol. wt. about 25000. CRLV acts as a helper for aphid transmission of carrot mottle virus (CMotV; ungrouped) from mixedly infected plants. Virus preparations purified from such plants possess the infectivity of both viruses but contain particles indistinguishable from those of CRLV; some of the particles are therefore thought to consist of CMotV RNA packaged in CRLV coat protein. When RNA from such preparations was electrophoresed in agarose/polyacrylamide gels, CMotV infectivity was associated with an RNA band that migrated ahead of the CRLV RNA band and had an estimated mol. wt. of about 1.5 x 106, similar to that previously found for the infective ssRNA extracted directly from Nicotiana clevelandii leaves infected with CMotV alone. Preparations of dsRNA from CMotV-infected N. clevelandii leaves contained two species: one of mol. wt. about 3.2 x 106, presumably the replicative form of the infective ssRNA, and the other, mol. wt. about 0.9 x 106, of unknown origin and function. The infective agent in buffer extracts of CMotV-infected N. clevelandii was resistant to RNase (although the enzyme acted as a reversible inhibitor of infection at high concentrations) and is therefore not unprotected RNA. It may be protected within the approximately 52 nm enveloped structures previously reported.

Development of the Rapid Assessment, Prioritisation and Referral Tool (RAPaRT) for multidisciplinary teams in emergency care settings

Objective This investigation utilised the expertise of allied members of multidisciplinary teams working in emergency care settings to develop and validate a Rapid Assessment Prioritisation and Referral Tool (RAPaRT). This instrument is intended for use among patients (with non-life threatening acuity) presenting to emergency care settings to indicate when referral to an allied member of the multidisciplinary team is warranted. Method This three stage instrument development and validation study included: a Delphi panel process to determine key criteria to guide instrument development and identify potential items to be carried forward for testing (stage 1); a prospective cohort of consecutive admissions (n=153) to investigate item sensitivity and specificity and retain only the most suitable items (stage 2); then final consultation with the Delphi panel to ensure the final instrument was clinically amenable (stage 3). Results 23 potential items were identified following stage 1. At the completion of item sensitivity and specificity analysis and in consultation with the Delphi panel, seven items were retained in the instrument. Area under the receiver operating characteristic curve was 0.803 for these seven items in predicting when a referral was warranted. Final consultation with the Delphi panel members also resulted in the addition of an open ended (eighth) item to allow description of any infrequent, but important, reason for referral. Conclusions The RAPaRT has demonstrated substantial promise as an efficient clinically amenable instrument to assist multidisciplinary teams in emergency care settings. Further research to investigate the wider implementation of the RAPaRT is warranted.

Re-presenting the past : authenticity and the historical novel

This project involved writing Turrwan (great man), a novel set in Queensland in the nineteenth century, and an investigation into the way historical novels portray the past. Turrwan tells the story of Tom Petrie, who was six when he arrived with his family at the notorious Moreton Bay Penal Colony in 1837. The thesis examines historical fiction as a genre with particular focus on notions of historical authenticity. It analyses the complexities involved in a non-Indigenous person writing about the Australian Aboriginal people, and reflects on the process of researching, planning and writing a historical novel.

Social networks in arid Australia : a review of concepts and evidence

Arid systems are markedly different from non-arid systems. This distinctiveness extends to arid-social networks, by which we mean social networks which are influenced by the suite of factors driving arid and semi-arid regions. Neither the process of how aridity interacts with social structure, nor what happens as a result of this interaction, is adequately understood. This paper postulates three relative characteristics which make arid-social networks distinct: that they are tightly bound, are hierarchical in structure and, hence, prone to power abuses, and contain a relatively higher proportion of weak links, making them reactive to crisis. These ideas were modified from workshop discussions during 2006. Although they are neither tested nor presented as strong beliefs, they are based on the anecdotal observations of arid-system scientists with many years of experience. This paper does not test the ideas, but rather examines them in the context of five arid-social network case studies with the aim of hypotheses building. Our cases are networks related to pastoralism, Aboriginal outstations, the ‘Far West Coast Aboriginal Enterprise Network’ and natural resources in both the Lake-Eyre basin and the Murray–Darling catchment. Our cases highlight that (1) social networks do not have clear boundaries, and that how participants perceive their network boundaries may differ from what network data imply, (2) although network structures are important determinants of system behaviour, the role of participants as individuals is still pivotal, (3) and while in certain arid cases weak links are engaged in crisis, the exact structure of all weak links in terms of how they place participants in relation to other communities is what matters.

NRM engagement between remote dryland communities and government agencies : success factors from Australia

The management of dryland environments involves the interaction of multiple government agencies and citizens, and is required to respond to a wide range of responsibilities and aspirations for a given region. This paper focuses on the characteristics of engagement between management agencies and citizens in a dryland region, presented here as a series of success factors. These factors are based on empirical research carried out in the Lake Eyre Basin in Australia, one of the world’s largest inwardly draining basins. The results reinforce generic and dryland-specific factors supporting successful community engagement. The former, such as building trust, working in partnership, supporting community champions, and maintaining transparency, are necessary but insufficient for achieving successful community engagement in the case study region. In addition, community engagement in the case study region also required respecting the extreme conditions and extraordinary variability of the Basin and committing to longer timeframes even if the outcomes of community engagement are slow to accrue, in order to take advantage of opportunities in more prosperous times.

Genuine community engagement in remote dryland regions : natural resource management in Lake Eyre Basin

Remote dryland regions are characterised by sparse populations and socially marginalised voices which pose particular challenges to natural resource management. This paper considers the issue of how to achieve community engagement in regions with these characteristics. In doing so, the paper contributes to an expanding international research agenda focusing on the distinct characteristics of arid and semi-arid regions under the heading of 'dryland syndrome'. The paper draws on government liaison officer and local community perspectives of successful engagement in the case-study region of Lake Eyre Basin, Australia. The results demonstrate that widely recognised characteristics of successful engagement are required but insufficient for genuine engagement in remote dryland regions. In addition to building trust through community ownership, being inclusive, effective communication, and adequate resources, genuine community engagement in drylands also requires respecting the extreme conditions and extraordinary variability of these areas. Residents of dryland regions seek genuine engagement yet engage opportunistically when seasons are conducive and when tangible outcomes are visible. © 2011 The Authors. Geographical Research © 2011 Institute of Australian Geographers.

Tools for successful NRM in the Lake Eyre Basin : achieving effective engagement

This chapter was developed as part of the ‘People, communities and economies of the Lake Eyre Basin’ project. It has been written for communities, government agencies and interface organisations involved in natural resource management (NRM) in the Lake Eyre Basin (LEB). Its purpose is to identify the key factors for successful community engagement processes relevant to the LEB and present tools and principles for successful engagement processes. The term ‘interface organisation’ is used here to refer to the diverse range of local and regional organisations (such as Catchment Committees or NRM Regional Bodies) that serve as linkages, or translators, between local communities and broader Australian and State Governments. The importance of fostering and harnessing effective processes of community engagement has been identified as crucial to building a prosperous future for rural and remote regions in Australia. The chapter presents an overview of the literature on successful community engagement processes for NRM, as well as an overview of the current NRM arrangements in the LEB. The main part of the chapter presents findings of the series of interviews conducted with the government liaison officers representing both state and federal organisations who are responsible for coordinating and facilitating regional NRM in the LEB, and with the members of communities of the LEB.

Terms of engagement : consensus or control in remote Australian resource management?

Public engagement is a defining feature of collaborative approaches to environmental management (Petts 2006, Whelan and Oliver 2005). Public engagement in this context is focused on incorporating residents and communities of interest in activities like ecological restoration, catchment management, and environmental conservation in a wide range of situations (Nelson and Pettit 2004, Petts 2007). Some authors consider public engagement to be a sign of healthy democratic functioning in society (Skocpol and Fiorina 1999). Others draw attention to overcoming widely noted practical limitations of top-down mechanisms, emphasising that public engagement results in programs being implemented more effectively (Broderick 2005, Leach et al. 1999).

Go with the flow : optimal AUV path planning in coastal environments

This paper describes a novel optimum path planning strategy for long duration AUV operations in environments with time-varying ocean currents. These currents can exceed the maximum achievable speed of the AUV, as well as temporally expose obstacles. In contrast to most other path planning strategies, paths have to be defined in time as well as space. The solution described here exploits ocean currents to achieve mission goals with minimal energy expenditure, or a tradeoff between mission time and required energy. The proposed algorithm uses a parallel swarm search as a means to reduce the susceptibility to large local minima on the complex cost surface. The performance of the optimisation algorithms is evaluated in simulation and experimentally with the Starbug AUV using a validated ocean model of Brisbane’s Moreton Bay.

Working knowledge for collaborative water planning in Australia���s Wet Tropics region

Cooperation between multiple environmental decision-makers and activities is necessary to address the impacts of diffuse sources of agricultural pollution on the water quality entering Australia���s Great Barrier Reef (GBR). Water planning efforts requires available knowledge to inform this co-operative water program implementation and reform. This paper uses knowledge sharing, translation and feedback features of collaboration as a way to assess knowledge work practices during key phases of the water planning process. This enabled a systematic review of knowledge work practices in partnership with collaborative water planning groups established to inform water quality program investment decisions in the GBR’s Wet Tropics region. This research builds on the growing academic and policy interest in the conditions required to enable different types of knowledge to be successfully used for policy-making by focusing on when, how and why knowledge work to meet these conditions is required.

Medieval knights amid the gum-trees : the experience of European heritage tourism in south-east Queensland, Australia

This paper is interested in the way in which the heritage of another place, time, and culture is repurposed for popular consumption in an experience economy, as well as the way in which the visitors experience their own past and the past of others. We trace the processes of engagement, education and nostalgia that occur when the European heritage is presented in a postcolonial context and an Australian environment. The information presented includes the results of qualitative and quantitative research conducted at the Abbey Museum over the December-Jan. period of 2012-13.

Working in partnership to develop engineering capability in energy efficiency

Energy efficiency is a complex topic to integrate into higher education curricula, with limited success internationally or in Australia. This paper discusses one of the successful initiatives within the Energy Efficiency Training Program, which was jointly managed and implemented by the New South Wales Office of Environment and Heritage and Department of Education and Communities. The state government initiative aimed to increase the knowledge and skills of the New South Wales workforce, help business to identify and implement energy efficiency projects, and provide professional development for the training providers. Key sectors targeted included property, construction, manufacturing and services. The Program was externally evaluated over the three years 2011 to 2013 and a range of insights were gained through these facilitated reflective opportunities, confirming and building upon literature on the topic to date. This paper presents lessons learned from the engineering part of the program (‘the project’), spanning government agencies, academic institutions, and academia. The paper begins with a contextual summary, followed by a synthesis of key learnings and implications for future training initiatives. It is intended that sharing these lessons will contribute to literature in the field, and assist other organisations in Australia and overseas planning similar initiatives.

Understanding the abnormal brain activity in epilepsy as a potential predictor of the onset of an epileptic seizure

The Brain Research Institute (BRI) uses various types of indirect measurements, including EEG and fMRI, to understand and assess brain activity and function. As well as the recovery of generic information about brain function, research also focuses on the utilisation of such data and understanding to study the initiation, dynamics, spread and suppression of epileptic seizures. To assist with the future focussing of this aspect of their research, the BRI asked the MISG 2010 participants to examine how the available EEG and fMRI data and current knowledge about epilepsy should be analysed and interpreted to yield an enhanced understanding about brain activity occurring before, at commencement of, during, and after a seizure. Though the deliberations of the study group were wide ranging in terms of the related matters considered and discussed, considerable progress was made with the following three aspects. (1) The science behind brain activity investigations depends crucially on the quality of the analysis and interpretation of, as well as the recovery of information from, EEG and fMRI measurements. A number of specific methodologies were discussed and formalised, including independent component analysis, principal component analysis, profile monitoring and change point analysis (hidden Markov modelling, time series analysis, discontinuity identification). (2) Even though EEG measurements accurately and very sensitively record the onset of an epileptic event or seizure, they are, from the perspective of understanding the internal initiation and localisation, of limited utility. They only record neuronal activity in the cortical (surface layer) neurons of the brain, which is a direct reflection of the type of electrical activity they have been designed to record. 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Working with Behavioural and Emotional Problems in Young People

This chapter outlines specific issues relating to behavioural and emotional problems in Aboriginal and Torres Strait Islander young people. It describes the most common disorders and their consequences, and how young Aboriginal people are at higher risk for developing such problems than other young Australians. The chapter also discusses the importance of psychosocial, cultural and environmental issues that need to be recognised in assessing and treating Aboriginal young people with behavioural and emotional problems. Issues concerning the delivery of both universal and culturally responsive prevention and intervention programs to address social and emotional wellbeing and mental health are discussed and possible interventions to enhance student engagement at school are provided. Finally, a range of mental health services for Aboriginal families which offer a culturally responsive approach to mental health treatment are listed.