Design and construction of an in-plant activation cassette for transgene expression and recombinant protein production in plants

Virus-based transgene expression systems have become particularly valuable for recombinant protein production in plants. The dual-module in-plant activation (INPACT) expression platform consists of a uniquely designed split-gene cassette incorporating the cis replication elements of Tobacco yellow dwarf geminivirus (TYDV) and an ethanol-inducible activation cassette encoding the TYDV Rep and RepA replication-associated proteins. The INPACT system is essentially tailored for recombinant protein production in stably transformed plants and provides both inducible and high-level transient transgene expression with the potential to be adapted to diverse crop species. The construction of a novel split-gene cassette, the inducible nature of the system and the ability to amplify transgene expression via rolling-circle replication differentiates this system from other DNA- and RNA-based virus vector systems used for stable or transient recombinant protein production in plants. Here we provide a detailed protocol describing the design and construction of a split-gene INPACT cassette, and we highlight factors that may influence optimal activation and amplification of gene expression in transgenic plants. By using Nicotiana tabacum, the protocol takes 6-9 months to complete, and recombinant proteins expressed using INPACT can accumulate to up to 10% of the leaf total soluble protein.

A History of Popular Indigenous Music

SINCE THE INVENTION OF recording technologies like the phonograph in the late 1800s, Indigenous music has been performed and recorded across Australia for a wide range of audiences. In the early twentieth century, for instance, music was recorded by anthropologists keen to capture the sounds of a culture that was believed to be in rapid decline (Thomas). Individual performers were not considered important in these recordings; their music was produced for scientific posterity rather than popular pleasure. And even though Aboriginal participation in local music festivals, touring vaudeville shows, and community gatherings was well documented throughout the twentieth century, it was not until the 1950s that Indigenous “pop stars” began to sell records for mass consumption(Dunbar-Hall and Gibson). Yet, with the persistence of recording artists like Jimmy Little over the past sixty years, Indigenous musicians have steadily gained prominence in Australia’s mainstream. This has been particularly true of the past twenty years, especially since the Sydney Olympics, where promotional strategies have brought about a new popular pride in musical achievements, based upon a celebrated history of diverse sounds and voices.

Evaluation of a pilot-scale oil extraction from microalgae for biodiesel production

Biodiesel derived from microalgae is one of a suite of potential solutions to meet the increasing demand for a renewable, carbon-neutral energy source. However, there are numerous challenges that must be addressed before algae biodiesel can become commercially viable. These challenges include the economic feasibility of harvesting and dewatering the biomass and the extraction of lipids and their conversion into biodiesel. Therefore, it is essential to find a suitable extraction process given these processes presently contribute significantly to the total production costs which, at this stage, inhibit the ability of biodiesel to compete financially with petroleum diesel. This study focuses on pilot-scale (100 kg dried microalgae) solvent extraction of lipids from microalgae and subsequent transesterification to biodiesel. Three different solvents (hexane, isopropanol (IPA) and hexane + IPA (1:1)) were used with two different extraction methods (static and Soxhlet) at bench-scale to find the most suitable solvent extraction process for the pilot-scale. The Soxhlet method extracted only 4.2% more lipid compared to the static method. However, the fatty acid profiles of different extraction methods with different solvents are similar, suggesting that none of the solvents or extraction processes were biased for extraction of particular fatty acids. Considering the cost and availability of the solvents, hexane was chosen for pilot-scale extraction using static extraction. At pilot-scale the lipid yield was found to be 20.3% of total biomass which is 2.5% less than from bench scale. Extracted fatty acids were dominated by polyunsaturated fatty acids (PUFAs) (68.94±0.17%) including 47.7±0.43 and 17.86±0.42% being docosahexaenoic acid (DHA) (C22:6) and docosapentaenoic acid (DPA) (C22:5, ω-3), respectively. These high amounts of long chain poly unsaturated fatty acids are unique to some marine microalgae and protists and vary with environmental conditions, culture age and nutrient status, as well as with cultivation process. Calculated physical and chemical properties of density, viscosity of transesterified fatty acid methyl esters (FAMEs) were within the limits of the biodiesel standard specifications as per ASTM D6751-2012 and EN 14214. The calculated cetane number was, however, significantly lower (17.8~18.6) compared to ASTM D6751-2012 or EN 14214-specified minimal requirements. We conclude that the obtained microalgal biodiesel would likely only be suitable for blending with petroleum diesel to a maximum of 5 to 20%.

A review on the production of levulinic acid and furanics from sugars

Sugarcane products represent an abundant and relatively low cost carbon resource that can be utilised to produce chemical intermediates such as levulinic acid and furanics. These chemicals can be easily upgraded to commodity and specialty chemicals and biofuels by high yielding and well established technologies. However, there are challenges and technical hurdles that need to be overcome before these chemical intermediates can be cost-effectively produced in commercial quantities. The paper reviews production of levulinic acid and furanics from sugars by homogeneous mineral acid catalysts, and reports on preliminary studies on the production of these compounds with environmentally friendly biodegradable sulfonic acids. The yields (>50% of theoretical) of levulinic acid, formic acid and furfural obtained with these organic acids are comparable to that of sulphuric acid currently used for their production.

Theory based production and project management

This paper gives an overview of an ongoing project endeavouring to advance theory-based production and project management, and the rationale for this approach is briefly justified. The status of the theoretical foundation of production management, project management and allied disciplines is discussed, with emphasis on metaphysical grounding of theories, as well as the nature of the heuristic solution method commonly used in these disciplines. Then, on-going work related to different aspects of production and project management is reviewed from both theoretical and practical orientation. Next, information systems agile project management is explored with a view to its re-use in generic project management. In production management, the consequences and implementation of a new, wider theoretical basis are analyzed. The theoretical implications and negative symptoms of the peculiarities of the construction industry for supply chains and supply chain management in construction are observed. Theoretical paths for improvements of inter-organisational relationships in construction which are fundamental for improvement of construction supply chains are described. To conclude, the observations made in this paper vis-à-vis production, project and supply chain management are related again to the theoretical basis of this paper, and finally directions for theory development and future research are given and discussed.

The design and development of music eScape: A new iPhone mood management music App

The physical, emotional, educational and social developmental challenges of adolescence can be associated with high levels of emotional vulnerability. Thus, the development of effective emotion-regulation strategies is crucial during this time period. Young people commonly use music to identify, express and regulate their emotions. Modern mobile technology provides an engaging, easily accessible means of assisting young people through music. A systematic contextual review identified 20 iPhone applications addressing emotions through music and two independent raters, using the Mobile App Rating Scale (MARS), evaluated the quality of the apps. Their characteristics, key features and overall quality will be presented. Three participatory design workshops (N=13, 6 males, 7 females; age 15-25) were conducted to explore young people’s use of music to enhance wellbeing. Young people were also asked to trial existing mood and music apps and to conceptualise their ultimate mood targeting music application. A thematic analysis of the participatory design workshops content identified the following music affect-regulation strategies: relationship building, modifying cognitions, modifying emotions, and immersing in emotions. The application of the key learnings from the mobile app review and participatory design workshops and the design and development of the music eScape app were presented and implications for future research was discussed.

'Music eScape': mHealth application

Research Background Young people’s avid use of mobile technologies in daily life has led to an increase in the design and research on mHealth (mobile health) interventions targeting young people. ‘Music eScape’ is a mobile based mood regulation app that uses an innovative approach to promoting young people’s wellbeing using music. Research Question The design, research, development and evaluation of ‘Music eScape’ addressed a number of research questions from across the fields of Psychology and Interactive and Visual Design. The specific design research question addressed was: How can interaction and visual design be utilized to promote and enable young people to effectively regulate their mood using music and how can the new design further promote their experience of empowerment, control and agency over actively directing their mood journey? Research Contribution Innovation and New Knowledge Through its unique visual interface design and interactivity, the application presents a novel approach to promoting young people’s wellbeing using music and a specific function that allows users to ‘draw’ their mood journey in order to generate a playlist. The mobile app is the first to contain a function that enables users to plan their mood journey and exercise a sense of agency, intentional choice and control over the mood shift and by extension, their wellbeing. The feature ‘drawing’ interface was designed by Oksana Zelenko using participatory design research and Russell’s circumplex model of affect (1980) to inform the key visual design concept and underpinning interaction design. Research Significance The significance of the design research component within the larger interdisciplinary practices that have informed ‘Music eScape’ (e.g. field of psychology, reported through journal articles and other related outcomes), is the unique visual and interactive presentation of participant data and music therapy research within the app interface and interaction design which improves and increases young people’s engagement with the health messages it contains. The industry quality standard is further demonstrated by the launch on Apple iTunes. This demonstrates the application meets the high professional requirements for national release and meets international standards. The app also creates a new benchmark for the quality of health apps on the market as it marks the industry release of a trialled evidence-based mHealth intervention co-designed with young people.

Community media production : access, institutions, and ethics

In policy terms, community media are known as the “third sector” of the media. The description reflects the historical expectation that community media can fulfill a need not met by the commercial and public service broadcasters. A defining element of this “need” has been the means to production for nonprofessionals, particularly groups not represented in the mainstream media. The historical construction of community media reveals production to be a guiding principle; both a means and an end in itself. This chapter examines the various rationales underpinning community media production, including empowerment, media diversity, and the independent producer movement. Using case studies from youth media, the chapter critiques producer-centric models of community media. In the contemporary media environment, production alone cannot meet the social needs that community media were established to address. Instead, I propose a rationale that combines both production and consumption ethics.

Contemporary cultures of service delivery to families : implications for music therapy

Family-centred and early intervention and prevention programs are a strong focus of current policy objectives within Australia, and a significant area of practice within the music therapy community. Recent shifts in the culture of policy and practice increasingly reflect ecological understandings by focussing on integrated and place-based approaches to service delivery. Further, current funding opportunities are strongly concerned with the extent to which interventions are able to reach out to highly vulnerable families that typically do not engage with services easily. Music therapy holds unique promise within these cultural shifts and thus advocates must develop a solid understanding of the concepts and related language in order to confidently engage with both funding and service systems. This paper uses an integrative review to first define and summarise current knowledge in three key areas relevant to contemporary Australian policy and practice: hard-to-reach families, home visiting as assertive outreach, and integrated or place-based service delivery. Evidence for the effectiveness of music therapy in relation to these key themes is then presented. Finally, the paper discusses the implications for the future of music therapy within the current Australian early intervention and prevention policy context and makes recommendations for moving forward on both practice and research fronts. While there is growing evidence and theory to suggest that music therapy may be uniquely efficacious in this area, greater Australian Journal of Music Therapy Vol 25, 2014 149 advocacy, documentation, research and adjustment of practices and language will further cement the position of the industry.

Creative Work Beyond the Creative Industries: An Introduction

Creative occupations exist across the entire economy. The creative worker’s habitus cannot be discovered by looking only in film studios, games companies or artist’s garrets. Work practices, evolved through the traditions of the creative and performing arts, are now deployed to create new services and products across all sectors, to develop process innovations, and to change the distribution thereof. Yet the bulk of academic study of creative work (both functionalist and critical), as well as the content of higher/further professional education programs and everyday understanding of creative workers, focuses on one subset of the Creative Industries: those involved in the production of cultural goods or services (film, television, music etc.) for consumption by the general public. And further, the bulk of existing academic work focuses on those creative workers employed in cultural production industries. However, as recent work has shown, this focus misses both the large (and increasing) number of creative workers embedded in industries beyond the core Creative Industries (for example, manufacturing, banking, mining) and those creative workers and firms that supply services to business as well as to the general public, such as architects, technical writers, and graphic designers (see Cunningham 2013; Potts and Cunningham 2008; Potts, Cunningham, Hartley and Omerod 2008). This book focuses on this subset of very important, and yet under-recognized creative workers: embedded creative workers and providers of creative services into other sectors of the economy, as indicated in the following taxonomy (Figure 1.1), which juxtaposes occupation and industry sector...

Association of MMP-2 activation potential with metastatic progression in human breast cancer cell lines independent of MMP-2 production

Background: Expression of matrix metalloproteinase-2 (MMP-2), the 72-kd type IV collagenase/gelatinase, by cancer cells has been implicated in metastasis through cancer cell invasion of basement membranes mediated by degradation of collagen IV. However, the abundance of this latent proenzyme in normal tissues and fluids suggests that MMP-2 proenzyme utilization is limited by its physiological activation rather than expression alone. We previously reported activation of this proenzyme by normal and malignant fibroblastoid cells cultured on collagen I (vitrogen) gels. Purpose: Our purposes in this study were 1) to determine whether MMP-2 activation is restricted to the more invasive human breast cancer cell lines and 2) to localize the activating mechanism. Methods: Zymography was used to monitor MMP-2 activation through detection of latent MMP-2 (72 kd) and mature species of smaller molecular weight (59 or 62 kd). Human breast cancer cell lines cultured on plastic, vitrogen, and other matrices were thus screened for MMP- 2 activation. Collagen I-cultured cells were exposed to cycloheximide, a protein synthesis inhibitor, or to protease inhibitors to determine the nature of the MMP-2-activating mechanism. Triton X-114 (TX-114) detergent extracts from cells cultured on collagen I or plastic were incubated with latent MMP-2 and analyzed by zymography to localize the MMP-2 activator. Results: MMP-2 activation was only induced by collagen I culture in the more aggressive, highly invasive estrogen receptor-negative, vimentin-positive human breast cancer cell lines (Hs578T, MDA-MB-436, BT549, MDA-MB-231, MDA- MB-435, MCF-7(ADR)) and was independent of MMP-2 production. MMP-2 activation was detected in cells cultured on collagen I gels but not in those cultured on gelatin gels, Matrigel, or thin layers of collagen I or IV, gelatin, or fibronectin. Collagen-induced activation was specific for the enzyme species MMP-2, since MMP-9, the 92-kd type IV collagenase/gelatinase, was not activatable under similar conditions. MMP-2 activation was inhibited by cycloheximide and was sensitive to a metalloproteinase inhibitor but not to aspartyl, serine, or cysteinyl protease inhibitors. MMP-2 activation was detected in the hydrophobic, plasma membrane-enriched, TX-114 extracts from invasive collagen I-cultured cells. Conclusion: Collagen I-induced MMP-2 activation is restricted to highly invasive estrogen receptor-negative, vimentin-positive human breast cancer cell lines, is independent of MMP-2 production, and is associated with metastatic potential. Our findings are consistent with plasma membrane localization of the activator. Implications: The MMP-2 activation mechanism may represent a new target for diagnosis, prognosis, and treatment of human breast cancer.

Production of levulinic acid and other chemicals from sugarcane fibre

Lignocellulosics represent a renewable resource for producing fuels and chemicals as an alternative to fossil resources. This study utilised an organic acid catalyst and a co-solvent to develop an environmentally friendly processing technology for the production of levulinic acid and furfural from a waste material, sugarcane fibre.

Disclosure of documents : production for inspection

In Altmann v Ioff of Victoria Friendly Society [2004] QDC 005 McGill DCJ considered the practical question in relation to disclosure of documents as to whether a party disclosing bundles of documents under UCPR r 217 was obliged to number or otherwise individually identify the documents

A broad-scale analysis of links between coastal fisheries production and mangrove extent : a case-study for northeastern Australia

The paradigm that mangroves are critical for sustaining production in coastal fisheries is widely accepted, but empirical evidence has been tenuous. This study showed that links between mangrove extent and coastal fisheries production could be detected for some species at a broad regional scale (1000s of kilometres) on the east coast of Queensland, Australia. The relationships between catch-per-unit-effort for different commercially caught species in four fisheries (trawl, line, net and pot fisheries) and mangrove characteristics, estimated from Landsat images were examined using multiple regression analyses. The species were categorised into three groups based on information on their life history characteristics, namely mangrove-related species (banana prawns Penaeus merguiensis, mud crabs Scylla serrata and barramundi Lates calcarifer), estuarine species (tiger prawns Penaeus esculentus and Penaeus semisulcatus, blue swimmer crabs Portunus pelagicus and blue threadfin Eleutheronema tetradactylum) and offshore species (coral trout Plectropomus spp.). For the mangrove-related species, mangrove characteristics such as area and perimeter accounted for most of the variation in the model; for the non-mangrove estuarine species, latitude was the dominant parameter but some mangrove characteristics (e.g. mangrove perimeter) also made significant contributions to the models. In contrast, for the offshore species, latitude was the dominant variable, with no contribution from mangrove characteristics. This study also identified that finer scale spatial data for the fisheries, to enable catch information to be attributed to a particular catchment, would help to improve our understanding of relationships between mangroves and fisheries production.