Preparation, Analysis and Use of an Affinity Adsorbent for the Purification of GST Fusion Protein

Methods are presented for the preparation, ligand density analysis and use of an affinity adsorbent for the purification of a glutathione S-transferase (GST) fusion protein in packed and expanded bed chromatographic processes. The protein is composed of GST fused to a zinc finger transcription factor (ZnF). Glutathione, the affinity ligand for GST purification, is covalently immobilized to a solid-phase adsorbent (Streamline™). The GST–ZnF fusion protein displays a dissociation constant of 0.6 x10-6 M to glutathione immobilized to Streamline™. Ligand density optimization, fusion protein elution conditions (pH and glutathione concentration) and ligand orientation are briefly discussed.

ICT in teacher education in the age of AITSL

This paper is drawn from a 2012-2013 OLT National Teaching Fellowship investigating the agencies impacting on whole-of course curriculum design in initial teacher education. The chief of these is AITSL (Australian Institute for Teaching and School Leadership) through the Australian Professional Standards for Teachers at Graduate level and the Program Accreditation Standards. This paper will begin with a discussion of the requirements on both beginning teachers and initial teacher education programs in regard to ICT (both content and pedagogy). It will then present case studies from four universities whose degree programs have been approved for implementation in 2014. It will focus on how each institution has responded to the APST as well as accreditation requirements. This will be based on responses to surveys to selected institutions and with one on one interviews to capture rich data. From this, it will draw a contemporary profile of how institutions are rising to the real requirements of ICT pedagogy within the regulatory constraints now in place. The methodology employed is qualitative and is based on document analysis enriched by interview data. It is important to know, as a profession, how future teachers are being introduced to and immersed in digital learning environments.

The land hoarding and land inspector dilemma in China : an evolutionary game theoretic perspective

China has experienced considerable economic growth since 1978, which was accompanied by unprecedented growth in urbanization and, more recently, by associated rising urban housing and land banking issues. One such issue is that of land hoarding - where real estate developers purchase land to hold unused in the rising market for a future lucrative sale, often several years later. This practice is outlawed in China, where land use is controlled by increasingly strengthened Government policies and inspectors. Despite this, land hoarding continues apace, with the main culprits being the developers and inspectors working subversively. This resembles a game between two players - the inspector and the developer - which provides the setting for this paper in developing an evolutionary game theory model to provide insights into dealing with the dilemmas faced by the players. The logic and dilemma of land banking strategy and illegal land banking issues are analysed, along with the land inspector’s role from a game theory perspective by determining the replication dynamic mechanism and evolutionary stable strategies under the various conditions that the players face. The major factors influencing the actions of land inspectors, on the other hand, are the costs of inspection, no matter if it is strict or indolent, conflict costs, and income and penalties from corruption. From this, it is shown that, when the net loss for corruption (income from corruption minus the penalties for corruption and cost of strict inspections) is less than the cost of strict inspections, the final evolutionary stable strategy of the inspectors is to carry out indolent inspections. Then, whether penalising developers for hoarding is severe or not, the evolutionary strategy for the developer is to hoard. The implications for land use control mechanisms and associated developer-inspector actions and counteractions are then examined in the light of the model's properties.

Learning through modelling in the primary years

Many nations are highlighting the need for a renaissance in the mathematical sciences as essential to the well-being of all citizens (e.g., Australian Academy of Science, 2006; 2010; The National Academies, 2009). Indeed, the first recommendation of The National Academies’ Rising Above the Storm (2007) was to vastly improve K–12 science and mathematics education. The subsequent report, Rising Above the Gathering Storm Two Years Later (2009), highlighted again the need to target mathematics and science from the earliest years of schooling: “It takes years or decades to build the capability to have a society that depends on science and technology . . . You need to generate the scientists and engineers, starting in elementary and middle school” (p. 9). Such pleas reflect the rapidly changing nature of problem solving and reasoning needed in today’s world, beyond the classroom. As The National Academies (2009) reported, “Today the problems are more complex than they were in the 1950s, and more global. They’ll require a new educated workforce, one that is more open, collaborative, and cross-disciplinary” (p. 19). The implications for the problem solving experiences we implement in schools are far-reaching. In this chapter, I consider problem solving and modelling in the primary school, beginning with the need to rethink the experiences we provide in the early years. I argue for a greater awareness of the learning potential of young children and the need to provide stimulating learning environments. I then focus on data modelling as a powerful means of advancing children’s statistical reasoning abilities, which they increasingly need as they navigate their data-drenched world.

Reduced intensity conditioning for allogeneic hematopoietic stem cell transplant determines the kinetics of acute Graft-Versus-Host Disease

Background Preparative myeloablative conditioning regimens for allogeneic hematopoietic stem-cell transplantation (HSCT) may control malignancy and facilitate engraftment but also contribute to transplant related mortality, cytokine release, and acute graft-versus-host disease (GVHD). Reduced intensity conditioning (RIC) regimens have decreased transplant related mortality but the incidence of acute GVHD, while delayed, remains unchanged. There are currently no in vivo allogeneic models of RIC HSCT, limiting studies into the mechanism behind RIC-associated GVHD. Methods We developed two RIC HSCT models that result in delayed onset GVHD (major histocompatibility complex mismatched (UBI-GFP/BL6 [H-2b]→BALB/c [H-2d]) and major histocompatibility complex matched, minor histocompatibility mismatched (UBI-GFP/BL6 [H-2b]→BALB.B [H-2b])) enabling the effect of RIC on chimerism, dendritic cell (DC) chimerism, and GVHD to be investigated. Results In contrast with myeloablative conditioning, we observed that RIC-associated delayed-onset GVHD is characterized by low production of tumor necrosis factor-α, maintenance of host DC, phenotypic DC activation, increased T-regulatory cell numbers, and a delayed emergence of activated donor DC. Furthermore, changes to the peritransplant milieu in the recipient after RIC lead to the altered activation of DC and the induction of T-regulatory responses. Reduced intensity conditioning recipients suffer less early damage to GVHD target organs. However, as donor cells engraft, activated donor DC and rising levels of tumor necrosis factor-α are associated with a later onset of severe GVHD. Conclusions Delineating the mechanisms underlying delayed onset GVHD in RIC HSCT recipients is vital to improve the prediction of disease onset and allow more targeted interventions for acute GVHD.

Cube Jam (Engagement Innovation Project)

Cube Jam is a project developed in response to the new and rising marketing in large-scale interactive public screens - the Cube being a premier site. Cube Jam will be a crossbreeding ‘think-ubator’ that rides on the back of the already nationally recognised Game On program and its digital communities. Via a bottom-up, non-directive approach Cube Jam will facilitate a series of design provocations within co-located Jam Studios; studios that are focused on supporting adaptation and new ideation and concept design. These Studios will seek new combinations of skills and knowledges with the intention of discovering provotypes of possibilities in both working and production methodologies and product outcomes.

Improving wound management for residents in Residential Aged Care Facilities: National dissemination and implementation of the evidence based Champions for Skin Integrity Program

Overview The incidence of skin tears, pressure injuries and chronic wounds increases with age [1-4] and therefore is a serious issue for staff and residents in Residential Aged Care Facilities (RACFs). A pilot project funded in Round 2 of the Encouraging Best Practice in Residential Aged Care (EBPRAC) program by the then Australian Government Department of Health and Ageing found that a substantial proportion of residents in aged care facilities experienced pressure injuries, skin tears or chronic wounds. It also found the implementation of the evidence based Champions for Skin Integrity (CSI) model of wound care was successful in significantly decreasing the prevalence and severity of wounds in residents, improving staff skills and knowledge of evidence based wound management, increasing staff confidence with wound management, increasing implementation of evidence based wound management and prevention strategies, and increasing staff awareness of their roles in evidence based wound care at all levels [5]. Importantly, during the project, the project team developed a resource kit on evidence based wound management. Two critical recommendations resulting from the project were that: - The CSI model or a similar strategic approach should be implemented in RACFs to facilitate the uptake of evidence based wound management and prevention - The resource kit on evidence based wound management should be made available to all Residential Aged Care Facilities and interested parties A proposal to disseminate or rollout the CSI model of wound care to all RACFs across Australia was submitted to the department in 2012. The department approved funding from the Aged Care Services Improvement Healthy Ageing Grant (ACSIHAG) at the same time as the Round 3 of the Encouraging Better Practice in Aged Care (EBPAC) program. The dissemination involved two crucial elements: 1. The updating, refining and distribution of a Champions for Skin Integrity Resource Kit, more commonly known as a CSI Resource Kit and 2. The presentation of intensive one day Promoting Healthy Skin “Train the Trainer” workshops in all capital cities and major regional towns across Australia Due to demand, the department agreed to fund a second round of workshops focussing on regional centres and the completion date was extended to accommodate the workshops. Later, the department also decided to host a departmental website for a number of clinical domains, including wound management, so that staff from the residential aged care sector had easy access to a central repository of helpful clinical resource material that could be used for improving the health and wellbeing of their older adults, consumers and carers. CSI Resource Kit Upgrade and Distribution: At the start of the project, a full evidence review was carried out on the material produced during the EBPRAC-CSI Stage 1 project and the relevant evidence based changes were made to the documentation. At the same time participants in the EBPRAC-CSI Stage 1 project were interviewed for advice on how to improve the resource material. Following this the documentation, included in the kit, was sent to independent experts for peer review. When this process was finalised, a learning designer and QUT’s Visual Communications Services were engaged to completely refine and update the design of the resources, and combined resource kit with the goal of keeping the overall size of the kit suitable for bookshelf mounting and the cost at reasonable levels. Both goals were achieved in that the kit is about the same size as a 25 mm A4 binder and costs between $19.00 and $28.00 per kit depending on the size of the print run. The dissemination of the updated CSI resource kit was an outstanding success. Demand for the kits was so great that a second print run of 2,000 kits was arranged on top of the initial print run of 4,000 kits. All RACFs across Australia were issued with a kit, some 2,740 in total. Since the initial distribution another 1,100 requests for kits has been fulfilled as well as 1,619 kits being distributed to participants at the Promoting Healthy Skin workshops. As the project was winding up a final request email was sent to all workshop participants asking if they required additional kits or resources to distribute the remaining kits and resources. This has resulted in requests for 200 additional kits and resources. Feedback from the residential aged care sector and other clinical providers who have interest in wound care has been very positive regarding the utility of the kit, (see Appendix 4). Promoting Healthy Skin Workshops The workshops also exceeded the project team’s initial objective. Our goal of providing workshop training for staff from one in four facilities and 450 participants was exceeded, with overwhelming demand for workshop places resulting in the need to provide a second round of workshops across Australia. At the completion of the second round, 37 workshops had been given, with 1286 participants, representing 835 facilities. A number of strategies were used to promote the workshops ranging from invitations included in the kit, to postcard mail-outs, broadcast emailing to all facilities and aged care networks and to articles and paid advertising in aged care journals. The most effective method, by far, was directly phoning the facilities. This enabled the caller to contact the relevant staff member and enlist their support for the workshop. As this is a labour intensive exercise, it was only used where numbers needed bolstering, with one venue rising from 3 registrants before the calls to 53 registrants after. The workshops were aimed at staff who had the interest and the capability of implementing evidence-based wound management within their facility or organisation. This targeting was successful in that a large proportion (68%) of participants were Registered Nurses, Nurse Managers, Educators or Consultants. Twenty percent were Endorsed Enrolled Nurses with the remaining 12% being made up of Personal Care Workers or Allied Health Professionals. To facilitate long term sustainability, the workshop employed train-the-trainer strategies. Feedback from the EBPRAC-CSI Stage 1 interviews was used in the development of workshop content. In addition, feedback from the workshop conducted at the end of the EBPRAC-CSI Stage 1 project suggested that change management and leadership training should be included in the workshops. The program was trialled in the first workshop conducted in Brisbane and then rolled out across Australia. Participants were asked to complete pre and post workshop surveys at the beginning and end of the workshop to determine how knowledge and confidence improved over the day. Results from the pre and post surveys showed significant improvements in the level of confidence in attendees’ ability to implement evidence based wound management. The results also indicated a significant increase in the level of confidence in ability to implement change within their facility or organisation. This is an important indication that the inclusion of change management/leadership training with clinical instruction can increase staff capacity and confidence in translating evidence into practice. To encourage the transfer of the evidence based content of the workshop into practice, participants were asked to prepare an Action Plan to be followed by a simple one page progress report three months after the workshop. These reports ranged from simple (e.g. skin moisturising to prevent skin tears), to complex implementation plans for introducing the CSI model across the whole organisation. Outcomes described in the project reports included decreased prevalence of skin tears, pressure injuries and chronic wounds, along with increased staff and resident knowledge and resident comfort. As stated above, some organisations prepared large, complex plans to roll out the CSI model across their organisation. These plans included a review of the organisation’s wound care system, policies and procedures, the creation of new processes, the education of staff and clients, uploading education and resource material onto internal electronic platforms and setting up formal review and evaluation processes. The CSI Resources have been enthusiastically sought and incorporated into multiple health care settings, including aged care, acute care, Medicare Local intranets (e.g. Map of Medicine e-pathways), primary health care, community and home care organisations, education providers and New Zealand aged and community health providers. Recommendations: Recommendations for RACFs, aged care and health service providers and government  Skin integrity and the evidence-practice gap in this area should be recognised as a major health issue for health service providers for older adults, with wounds experienced by up to 50% of residents in aged care settings (Edwards et al. 2010). Implementation of evidence based wound care through the Champions for Skin Integrity model in this and the pilot project has demonstrated the prevalence of wounds, wound healing times and wound infections can be halved.  A national program and Centre for Evidence Based Wound Management should be established to: - expand the reach of the model to other aged care facilities and health service providers for older adults - sustain the uptake of models such as the Champions for Skin Integrity (CSI) model - ensure current resources, expertise and training are available for consumers and health care professionals to promote skin integrity for all older adults  Evidence based resources for the CSI program and similar projects should be reviewed and updated every 3 – 4 years as per NH&MRC recommendations  Leadership and change management training is fundamental to increasing staff capacity, at all levels, to promote within-organisation dissemination of skills and knowledge gained from projects providing evidence based training Recommendations for future national dissemination projects  A formal program of opportunities for small groups of like projects to share information and resources, coordinate activities and synergise education programs interactively would benefit future national dissemination projects - Future workshop programs could explore an incentive program to optimise attendance and reduce ‘no shows’ - Future projects should build in the capacity and funding for increased follow-up with workshop attendees, to explore the reasons behind those who are unable to translate workshop learnings into the workplace and identify factors to address these barriers.

Product stability and sequestration mechanisms in Solanum tuberosum engineered to biosynthesize high value ketocarotenoids

To produce commercially valuable ketocarotenoids in Solanum tuberosum, the 4, 4′ β-oxygenase (crtW) and 3, 3′ β-hydroxylase (crtZ) genes from Brevundimonas spp. have been expressed in the plant host under constitutive transcriptional control. The CRTW and CRTZ enzymes are capable of modifying endogenous plant carotenoids to form a range of hydroxylated and ketolated derivatives. The host (cv. Désirée) produced significant levels of nonendogenous carotenoid products in all tissues, but at the apparent expense of the economically critical metabolite, starch. Carotenoid levels increased in both wild-type and transgenic tubers following cold storage; however, stability during heat processing varied between compounds. Subcellular fractionation of leaf tissues revealed the presence of ketocarotenoids in thylakoid membranes, but not predominantly in the photosynthetic complexes. A dramatic increase in the carotenoid content of plastoglobuli was determined. These findings were corroborated by microscopic analysis of chloroplasts. In tuber tissues, esterified carotenoids, representing 13% of the total pigment found in wild-type extracts, were sequestered in plastoglobuli. In the transgenic tubers, this proportion increased to 45%, with esterified nonendogenous carotenoids in place of endogenous compounds. Conversely, nonesterified carotenoids in both wild-type and transgenic tuber tissues were associated with amyloplast membranes and starch granules.

Environment, political economies and livelihood change

Between the national and household factors, community or “meso-level” changes in political economy and livelihoods in southwestern Bangladesh illustrate that in order to understand the impacts on people and nations of climate change-related environmental changes – changes that are expected to include rising sea level, saline inundation, and increased likelihood and intensity of cyclones in Bangladesh – we need to understand the dynamics of the built and natural environment and the political economies these sustain. Meso-level political economies affect the sources of income and livelihood available in distressed environmental conditions, and therefore influence how well the people in them can adapt to changing environmental conditions. In this study we have seen the underlying political economies whose dynamics, and not slow onset environmental changes or disastrous environmental events, are pushing Bangladeshis to incorporate migration strategies into their livelihood strategies.

Transforming pedagogy: Building the new ‘old school’

Global shifts in design practice, design education and in higher education in general have radically changed teaching practices over the past decade. In a time of tightening budgets and other factors, various paradigm shifts and challenges are continually evolving and rapidly changing the higher education landscape. Rising to these challenges and responding to them is a complex task, requiring creative approaches applied to traditional methods and tools in interior architecture and interior design courses. The focus of this paper is a discussion of the adaptation of an ‘old school’ traditional tutorial approach within the context of online interior design education, which was applied in an endeavour to increase student engagement with current relevant literature pertinent to interior design theory and practice. The adaptation and application of this tutorial approach, the O-tutorial, creates a necessary link to critical thinking and meta-cognition connecting theoretical concepts to interior design process and practice. Initial data reveals that the O-tutorial allows students to engage and critically analyse issues surrounding theory and practice, thus equipping them with the skills as future design professionals. The paper concludes with reflections and recommendations for interior design education and future potential use and application of the O-tutorial.

Where do ED patients come from?

A lack of access to primary care services, decreasing numbers of general practitioners (GPs) and free of charge visits have been cited as factors contributing to the rising demand on emergency departments. This study aims to investigate the sources of patients' referrals to emergency departments and track changes in the source of referral over a six-year period in Queensland. Data from Queensland Emergency Departments Information Systems were analyzed based on records from 21 hospitals for the periods 2003–04 to 2008–09. The emergency department data were compared with publicly available data on GPs services and patients attendance rates. In Queensland, the majority of patients are self-referred and a 6.6% growth between 2003–04 and 2008–09 (84.4% to 90% respectively) has been observed. The number of referrals made by GPs, hospitals and community services decreased by 29.4%, 40%, 42% respectively during the six-year period. The full-time workload equivalent GPs per 100,000 people increased by 4.5% and the number of GP attendances measured per capita rose by 4% (4.25 to 4.42). An examination of changes in the triage category of self-referred patients revealed an increase in triage category 1-3 by 60%, 36.2%, and 14.4% respectively. The number of self-referred patients in triage categories 4–5 decreased by 10.5% and 21.9% respectively. The results of this analysis reveal that although the number of services provided by GPs increased, the amount of referrals decreased, and the proportion of self-referred patients to emergency departments rose during the six-year period. In addition, a growth in urgent triage categories (1–3) has been observed, with a decline in the number of non-urgent categories (4–5) among patients who came directly to emergency departments. Understanding the reasons behind this situation is crucial for appropriate demand management. Possible explanations will be sought and presented based on patients' responses to an emergency department users' questionnaire.

Why does child labor persist with declining poverty?

We develop a dynamic overlapping generations model to highlight the role of income inequality in explaining the persistence of child labor under declining poverty. Differential investment in two forms of human capital—schooling and health—in the presence of inequality gives rise to a nonconvergent income distribution in the steady state characterized by multiple steady states of relative income with varying levels of education, health, and child labor. The child labor trap thus generated is shown to preserve itself despite rising per capita income. Policy recommendations include public provision of education targeted toward reducing schooling costs for the poor or raising the efficacy of public health infrastructure.

Re-thinking sustainable solutions: Innovation inspired by nature

Australian rural landscapes are facing a crisis from land degradation due to rising salinity levels, soil acidification and soil erosion. There is growing consensus amongst the businesses community, government departments and research organisations that the real solution to these problems and the broader sustainability dilemma comes by taking a ‘whole of system’ approach to agricultural and rangelands management. This article introduces two cutting-edge concepts – Biomimicry and Natural Sequence Farming – to illustrate how whole-system thinking can effectively and profitably address the challenges facing agriculture and rangelands.

Multiple sulfur isotope analyses support a magmatic model for the volcanogenic massive sulfide deposits of the Teutonic Bore volcanic complex, Yilgarn Craton, Western Australia

We report sensitive high mass resolution ion microprobe, stable isotopes (SHRIMP SI) multiple sulfur isotope analyses (32S, 33S, 34S) to constrain the sources of sulfur in three Archean VMS deposits—Teutonic Bore, Bentley, and Jaguar—from the Teutonic Bore volcanic complex of the Yilgarn Craton, Western Australia, together with sedimentary pyrites from associated black shales and interpillow pyrites. The pyrites from VMS mineralization are dominated by mantle sulfur but include a small amount of slightly negative mass-independent fractionation (MIF) anomalies, whereas sulfur from the pyrites in the sedimentary rocks has pronounced positive MIF, with ∆33S values that lie between 0.19 and 6.20‰ (with one outlier at −1.62‰). The wall rocks to the mineralization include sedimentary rocks that have contributed no detectable positive MIF sulfur to the VMS deposits, which is difficult to reconcile with the leaching model for the formation of these deposits. The sulfur isotope data are best explained by mixing between sulfur derived from a magmatic-hydrothermal fluid and seawater sulfur as represented by the interpillow pyrites. The massive sulfide lens pyrites have a weighted mean ∆33S value of −0.27 ± 0.05‰ (MSWD = 1.6) nearly identical with −0.31 ± 0.08‰ (MSWD = 2.4) for pyrites from the stringer zone, which requires mixing to have occurred below the sea floor. We employed a two-component mixing model to estimate the contribution of seawater sulfur to the total sulfur budget of the two Teutonic Bore volcanic complex VMS deposits. The results are 15 to 18% for both Teutonic Bore and Bentley, much higher than the 3% obtained by Jamieson et al. (2013) for the giant Kidd Creek deposit. Similar calculations, carried out for other Neoarchean VMS deposits give value between 2% and 30%, which are similar to modern hydrothermal VMS deposits. We suggest that multiple sulfur isotope analyses may be used to predict the size of Archean VMS deposits and to provide a vector to ore deposit but further studies are needed to test these suggestions.