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.

Plasmid DNA purification via the use of a dual affinity protein

Methods are presented for the production, affinity purification and analysis of plasmid DNA (pDNA). Batch fermentation is used for the production of the pDNA, and expanded bed chromatography, via the use of a dual affinity glutathione S-transferase (GST) fusion protein, is used for the capture and purification of the pDNA. The protein is composed of GST, which displays affinity for glutathione immobilized to a solid-phase adsorbent, fused to a zinc finger transcription factor, which displays affinity for a target 9-base pair sequence contained within the target pDNA. A Picogreen™ fluorescence assay and/or anx ethidium bromide agarose gel electrophoresis assay can be used to analyze the eluted pDNA.

How does the use of a reflective journal enhance students’ critical thinking about complexity?

Introduction to Youth Services is a second year Social Work and Human Services unit. In this unit a reflective writing task was introduced to assess students’ reflections on an ongoing tutorial discussion to which they contributed. The discussion was based on a fictional young person each tutorial group ‘worked with’ across eight weeks of a semester. In developing the process and the criteria for the reflective journal, the ideas raised by the Teaching and Assessing Reflective Learning (TARL) in Higher Education project (see Chap. 2) were utilised, scaffolding the work with resources and submission of a draft. The students were also invited to choose the form of reflective process they used, it could be a written journal but did not need to be. The evidence exemplified that a reflective journal is an effective tool for students to record their developing understanding regarding the concept that issues people experience are complex and compounding. Importantly, it was also a useful vehicle for students to begin to consider the impacts of their own and others’ values and beliefs on their response to the issues raised within the case discussion. The reflective journal also helped participants to consider how this learning contributes to the ongoing development of their professional practice framework.

Clinical use of diodes and micro-chambers to obtain accurate small field output factor measurements

There have been substantial advances in small field dosimetry techniques and technologies, over the last decade, which have dramatically improved the achievable accuracy of small field dose measurements. This educational note aims to help radiation oncology medical physicists to apply some of these advances in clinical practice. The evaluation of a set of small field output factors (total scatter factors) is used to exemplify a detailed measurement and simulation procedure and as a basis for discussing the possible effects of simplifying that procedure. Field output factors were measured with an unshielded diode and a micro-ionisation chamber, at the centre of a set of square fields defined by a micro-multileaf collimator. Nominal field sizes investigated ranged from 6×6 to 98×98 mm2. Diode measurements in fields smaller than 30 mm across were corrected using response factors calculated using Monte Carlo simulations of the full diode geometry and daisy-chained to match micro-chamber measurements at intermediate field sizes. Diode measurements in fields smaller than 15 mm across were repeated twelve times over three separate measurement sessions, to evaluate the to evaluate the reproducibility of the radiation field size and its correspondence with the nominal field size. The five readings that contributed to each measurement on each day varied by up to 0.26%, for the “very small” fields smaller than 15 mm, and 0.18% for the fields larger than 15 mm. The diode response factors calculated for the unshielded diode agreed with previously published results, within 1.6%. The measured dimensions of the very small fields differed by up to 0.3 mm, across the different measurement sessions, contributing an uncertainty of up to 1.2% to the very small field output factors. The overall uncertainties in the field output factors were 1.8% for the very small fields and 1.1% for the fields larger than 15 mm across. Recommended steps for acquiring small field output factor measurements for use in radiotherapy treatment planning system beam configuration data are provided.

On the use of optical flow for scene change detection and description

We propose the use of optical flow information as a method for detecting and describing changes in the environment, from the perspective of a mobile camera. We analyze the characteristics of the optical flow signal and demonstrate how robust flow vectors can be generated and used for the detection of depth discontinuities and appearance changes at key locations. To successfully achieve this task, a full discussion on camera positioning, distortion compensation, noise filtering, and parameter estimation is presented. We then extract statistical attributes from the flow signal to describe the location of the scene changes. We also employ clustering and dominant shape of vectors to increase the descriptiveness. Once a database of nodes (where a node is a detected scene change) and their corresponding flow features is created, matching can be performed whenever nodes are encountered, such that topological localization can be achieved. We retrieve the most likely node according to the Mahalanobis and Chi-square distances between the current frame and the database. The results illustrate the applicability of the technique for detecting and describing scene changes in diverse lighting conditions, considering indoor and outdoor environments and different robot platforms.

Green: Exploring the aestheticized use of chroma-key techniques and technologies in two intermedial productions

Peggy Shaw’s RUFF, (USA 2013) and Queensland Theatre Company’s collaboration with Queensland University of Technology, Total Dik!, (Australia 2013) overtly and evocatively draw on an aestheticized use of the cinematic techniques and technologies of Chroma Key to reveal the tensions in their production and add layers to their performances. In doing so they offer invaluable insight where the filmic and theatrical approaches overlap. This paper draws on Eckersall, Grehan and Scheer’s New Media Dramaturgy (2014) to reposition the frame as a contribution to intermedial theatre and performance practices in light of increasing convergence between seemingly disparate discourses. In RUFF, the scenic environment replicates a chroma-key ‘studio’ which facilitates the reconstruction of memory displaced after a stroke. RUFF uses the screen and projections to recall crooners, lounge singers, movie stars, rock and roll bands, and an eclectic line of eccentric family members living inside Shaw. While the show pays tribute to those who have kept her company across decades of theatrical performance, use of non-composited chroma-key technique as a theatrical device and the work’s taciturn revelation of the production process during performance, play a central role in its exploration of the juxtaposition between its reconstructed form and content. In contrast Total Dik! uses real-time green screen compositing during performance as a scenic device. Actors manipulate scale models, refocus cameras and generate scenes within scenes in the construction of the work’s examination of an isolated Dictator. The ‘studio’ is again replicated as a site for (re)construction, only in this case Total Dik! actively seeks to reveal the process of production as the performance plays out. Building on RUFF, and other works such as By the Way, Meet Vera Stark, (2012) and Hotel Modern’s God’s Beard (2012), this work blends a convergence of mobile technologies, models, and green screen capture to explore aspects of transmedia storytelling in a theatrical environment (Jenkins, 2009, 2013). When a green screen is placed on stage, it reads at once as metaphor and challenge to the language of theatre. It becomes, or rather acts, as a ‘sign’ that alludes to the nature of the reconstructed, recomposited, manipulated and controlled. In RUFF and in Total Dik!, it is also a place where as a mode of production and subsequent reveal, it adds weight to performance. These works are informed by Auslander (1999) and Giesenkam (2007) and speak to and echo Lehmann’s Postdramatic Theatre (2006). This paper’s consideration of the integration of studio technique and live performance as a dynamic approach to multi-layered theatrical production develops our understanding of their combinatory use in a live performance environment.

Use of parametric modelling and climate-based metrics for the efficient design of daylight strategies in buildings with complex geometries

The purpose of this research is to assess daylight performance of buildings with climatic responsive envelopes with complex geometry that integrates shading devices in the façade. To this end two case studies are chosen due to their complex geometries and integrated daylight devices. The effect of different parameters of the daylight devices is analysed through Climate base daylight metrics.

The use of ecosystem services valuation in Australian coastal zone management

The use and influence of ecosystem services valuation in management decision-making, particularly as it relates to coastal zone management, remains largely unexplored in the academic literature. A recent Australia-wide survey of decision-makers involved in coastal zone management examined if, how and to what extent economic valuation of coastal and marine ecosystem services is used in, and influences, decision-making in Australia. The survey also identified a set of cases where economic valuation of ecosystem services was used for decision-making, and reasons why economic values may or may not be considered in the decision-making process. This paper details the method and results from this survey. Overall, there is strong empirical evidence that economic valuation of ecosystem services is used, but with important variation across coastal and marine management contexts. However, the impact of ecosystem services valuation on policy appears to be globally weak.

Quality assurance of assessment and moderation discourses involving sessional staff

Quality assurance is a major agenda in tertiary education. The casualisation of academic work, especially in teaching, is also a quality assurance issue. Casual or sessional staff members teach and assess more than 50% of all university courses in Australia, and yet the research in relation to the role sessional staff play in quality assurance of student assessment outcomes is scarce. Moderation processes are a pivotal part of robust quality assurance measures. Drawing upon previous work surrounding four discourses of moderation, this pilot project reports the results of research into the role and impact of sessional staff in moderation processes at tertiary level. Qualitative data were gathered through focus interviews. Results, in the form of various moderation discourses, indicate that sessional staff impact the formal quality assurance processes in numerous ways.

Too much knowledge for a nurse? Use of physical assessment by final-semester nursing students

There is debate around the scope of physical assessment skills that should be taught in undergraduate nursing programs. Yet this debate is largely uninformed by evidence on what is learned and practiced by nursing students. This study examined the pattern and correlates of physical assessment skill utilization by 208 graduating nursing students at an Australian university, including measures of knowledge, frequency of use and perceived barriers to physical assessment skills during clinical practice. Of the 126 skills surveyed, on average only five were used every time students practiced. Core skills reflected inspection or general observation of the patient; none involved complex palpation, percussion or auscultation. Skill utilization was also shaped by specialty area. Most skills (70%) were, on average, never performed or learned and students perceived nursing physical assessment was marginalized in both university and workplace contexts. Lack of confidence was thus a significant barrier to use of skills. Based on these findings we argue that the current debate must shift to how we might best support students to integrate comprehensive physical assessment into nursing practice.

Analysis and evaluation of use of the V1 and South East Freeway Bicycleway post completion of Stage C of the V1

Executive Summary: Completion of the Veloway 1 (V1) will provide a dedicated and safe route for cyclists between the Brisbane CBD and the Gateway Motorway off-ramp at Eight Mile Plains alongside the South East Motorway. The V1 is being delivered in stages and when completed will provide a dedicated 3m wide cycleway 17km in length. Two stages (D and E) remain to be constructed to complete the V1. Major trip attractors along the V1 include the Mater, Princes Alexandra and Greenslopes Hospitals, two campuses of Griffith University, Garden City shopping centre and the Australian Tax Office. This report assesses the available evidence on the impacts on cycling behaviour of the recently completed V1 Stage C. The data sources informing this review include three intercept surveys, motion activated traffic cameras and travel time surveys on the V1 and adjoining South East Freeway Bikeway (SEFB), Strava app data, and cyclist crash data along Logan Road. The key findings from the evidence are that the completed V1 Stage C has: a Attracted cyclists from Holland Park, Holland Park West, Mt Gravatt and southern parts of Tarragindi onto the V1 Stage C. b Reduced the crash exposure of pedestrians to cyclists by attracting higher speed cyclists off the adjoining SEFB onto the cycling dedicated V1 Stage C. c Reduced the potential crash exposure of cyclists to motor vehicles by attracting cyclists off Logan Road on to the V1. d Provided travel time benefits to cyclists and reduced road crossings (eight down to two). e Predominantly attracted adults commuting alone to and from work and university. The evidence shows that the two traffic crossings across Birdwood Road (required as a temporary measure until the V1 is completed) negate much of the travel time gains of the V1 Stage C compared to the adjoining SEFB for southbound cyclists. Many cyclists accessing the V1 Stage C from the south are cycling in high-volume vehicular traffic lanes to reduce their travel time along Birdwood Road, but in the process are increasing their exposure to crashes with motor vehicles. Based on these findings this report recommends that TMR: a. Continue with plans to complete the V1 Veloway b. Undertake an engineering feasibility assessment to determine the viability of constructing a section of the V1 Stage E from the intersection Weller and Birdwood Roads over Marshall Road and along Bapaume Road on the western side of the Motorway to the intersection of Bapaume and Sterculia Roads. c. In the interim, improve signage and Birdwood Road crossing points for cyclists accessing and egressing the southern end of the V1 Stage C. d. Work with Brisbane City Council to identify the safest and most practical bicycle facilities to facilitate cycle travel between Logan Road and the V1 south of Birdwood Road. e. Improve the awareness of the V1 Stage C through signage for cyclists approaching from the north with the aim of providing a better understanding of the route of the V1 to the south. f. Refine the use of motion activated traffic cameras to improve the capture rate of useable images and obtain an ongoing collection over time of V1 usage data. g. Undertake discussions with Strava, Inc. to refine the presentation of Strava data to improve visual understanding of maps showing before and after cycle route volumes along and on roads leading to the V1.

Ants, apples and the ABCs: The use of commercial phonics programmes in prior-to-school children's services

Commercial phonics programmes (e.g. Jolly Phonics and Letterland) are becoming widely used in the early years of school. These programmes claim to use a systematic explicit approach, considered as the preferred method of phonics instruction for teaching alphabetic code-breaking skills in Australia and the UK in the first years of school (Department of Education, Science and Training, 2005; Rose, 2006). However, little is known about the extent to which they are being used in prior-to-school settings, and the reasons behind decisions to use them. This study surveyed 283 early childhood staff in Sydney, Australia and found that commercial phonics programmes were being used in 36% of the early childhood settings surveyed. Staff with early childhood univer- sity qualifications and staff working in not-for-profit service types were less likely to use a commercial phonics programme than staff without university qualifications and staff working in for-profit services. Staff with less than 10 years’ experience were also more likely to use a commercial phonics programme. The rationale behind decisions deter- mining whether or not staff used the programmes ranged from pragmatic reasons, such as parent pressure or higher management decisions, to pedagogical reasons, such as teacher beliefs about how children learn to read and write. The practices staff engage in to teach phonics are explored.

How European public sector agencies innovate: The use of bottom-up, policy-dependent and knowledge-scanning innovation methods

Factor and cluster analysis are used to identify different methods that public sector agencies in Europeuse to innovate, based on data from a 2010 survey of 3273 agencies. The analyses identify three types ofinnovative agencies: bottom-up, knowledge-scanning, and policy-dependent. The distribution of bottom-up agencies across European countries is positively correlated with average per capita incomes while thedistribution of knowledge-scanning agencies is negatively correlated with income. In contrast, there isno consistent pattern by country in the distribution of policy-dependent agencies. Regression resultsthat control for agency characteristics find that innovation methods are significantly correlated with thebeneficial outcomes of innovation, with bottom-up and knowledge-scanning agencies out-performingpolicy-dependent agencies.

A tsunami of unmet needs: Pancreatic and ampullary cancer patients’ supportive care needs and use of community and allied health services

Objective People diagnosed with pancreatic cancer have the worst survival prognosis of any cancer. No previous research has documented the supportive care needs of this population. Our objective was to describe people’s needs and use of support services and to examine whether these differed according to whether or not patients had undergone surgical resection. Methods Queensland pancreatic or ampullary cancer patients (n=136, 54% of those eligible) completed a survey which assessed 34 needs across 5 domains (SCNS-SF34) and use of health services. Differences by resection were compared with Chi-squared tests. Results Overall, 96% of participants reported having some needs. More than half reported moderate-to-high unmet physical (54%) or psychological (52%) needs whereas, health system/information (32%), patient care (21%) and sexuality needs (16%) were described less frequently. The three most frequently reported moderate-to-high needs included ‘not being able to do things they used to do’ (41%), ‘concerns about the worries of those close’ (37%), and ‘uncertainty about the future’ (30%). Patients with non-resectable disease reported greater individual information needs but their needs were otherwise similar to patients with resectable disease. Self-reported use of support was low; only 35% accessed information, 28%, 18% and 15% consulted a dietician, complementary medicine practitioner or mental health practitioner, respectively. Palliative care access was greater (59% vs 27%) among those with non-resectable disease. Conclusion Very high levels of needs were reported by people with pancreatic or ampullary cancer. Future work needs to elucidate why uptake of appropriate supportive care is low and which services are required.