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.

Kids, counselors and troubles-telling : morality-in-action in talk on an Australian children’s helpline

This article begins with the premise that morality is an intrinsic, although often invisible, aspect of everyday social action. Drawn from a corpus of fifty audiorecorded telephone calls to Kids Helpline, an Australian helpline for children and young people, we examine one call to show how the young caller and counsellor co-construct ‘morality-in-action’. Ethnomethodological understandings and, in particular, Sacks’ (1992) description of ‘Class 2’ rules and infractions show how an adolescent caller and counsellor collaboratively assemble moral versions of the caller. In puzzling out possible motives, the caller and counsellor can be seen to be attending to the implications of different moral versions of the caller. This attribution of motives is moral work in action, with motives contingently assembled, displayed and evaluated, with such work understood as displays of moral reasoning. The counselling call makes visible the counsellor’s interactional work to support and empower the client. Analysis such as this offers counsellors ways of understanding and making visible their interactional and moral work within helpline call interactions.

Sectional analysis of stormwater treatment performance of a constructed wetland

Constructed wetlands are among the most common Water Sensitive Urban Design (WSUD) measures for stormwater treatment. These systems have been extensively studied to understand their performance and influential treatment processes. Unfortunately, most past studies have been undertaken considering a wetland system as a lumped system with a primary focus on the reduction of the event mean concentration (EMC) values of specific pollutant species or total pollutant load removal. This research study adopted an innovative approach by partitioning the inflow runoff hydrograph and then investigating treatment performance in each partition and their relationships with a range of hydraulic factors. The study outcomes confirmed that influenced by rainfall characteristics, the constructed wetland displays different treatment characteristics for the initial and later sectors of the runoff hydrograph. The treatment of small rainfall events (<15 mm) is comparatively better at the beginning of runoff events while the trends in pollutant load reductions for large rainfall events (>15 mm) are generally lower at the beginning and gradually increase towards the end of rainfall events. This highlights the importance of ensuring that the inflow into a constructed wetland has low turbulence in order to achieve consistent treatment performance for both, small and large rainfall events.

Staged homicides: An examination of common features of faked burglaries, suicides, accidents and car accidents

A staged crime scene involves deliberate alteration of evidence by the offender to simulate events that did not occur for the purpose of misleading authorities (Geberth, 2006; Turvey, 2000). This study examined 115 staged homicides from the USA to determine common elements; victim and perpetrator characteristics; and specific features of different types of staged scenes. General characteristics include: multiple victims and offenders; a previous relationship be- tween parties involved; and victims discovered in their own home, often by the offender. Staged scenes were separated by type with staged burglaries, suicides, accidents, and car accidents examined in more detail. Each type of scene displays differently with separate indicators and common features. Features of staged burglaries were: no points of entry/exit staged; non-valuables taken; scene ransacking; offender self- injury; and offenders bringing weapons to the scene. Features of staged suicides included: weapon arrangement and simulating self-injury to the victim; rearranging the body; and removing valuables. Examples of elements of staged accidents were arranging the implement/weapon and re- positioning the deceased; while staged car accidents involved: transporting the body to the vehicle and arranging both; mutilation after death; attempts to secure an alibi; and clean up at the primary crime scene. The results suggest few staging behaviors are used, despite the credibility they may have offered the façade. This is the first peer-reviewed, published study to examine the specific features of these scenes, and is the largest sample studied to date.

'I wonder what you know...': Teachers designing requests for factual information

The early years are significant in optimising children’s educational, emotional and social outcomes and have become a major international policy priority. Within Australia, policy levers have prioritised early childhood education, with a focus on program quality, as it is associated with lifelong success. Longitudinal studies have found that high quality teacher-child interactions are an essential element of high quality programs, and teacher questioning is one aspect of teacher-child interactions that has been attributed to affecting the quality of education, linking open ended questioning to higher cognitive achievement. Teachers, however, overwhelmingly ask more closed than open questions. In the classroom, like everyday interaction, questions in interaction require answers. They are used to request, offer, repair, challenge, seek agreement (Curl & Drew, 2008; Enfield, Stivers, & Levinson, 2010; Hayano, 2013; Schegloff, 2007). Teachers use questions to set agendas and manage lessons (McHoul, 1978; Mehan, 1979; Sacks, 1995), and to gauge students’ knowledge and understanding (Lerner, 1995; McHoul, 1978; Mehan, 1979). Drawing on data from the Australian Research Council project Interacting with Knowledge: Interacting with people: Web searching in early childhood, this paper focuses on an extended sequence of talk between a teacher with two students aged between 3.5 and 5 years in a preschool classroom. The episode, drawn from a corpus of over 200 hours of video recorded data, captures how the teacher and children undertake an online search for images of lady beetles and hairy caterpillars on the Web. Ethnomethodological and conversation analysis approaches examine how the teacher asks questions, which call on the children to display their factual knowledge about the search topic. The fine grained analysis shows how teachers design their interactions to prompt children’s displays of factual knowledge, and how the design of factual questions affect a student’s response in terms of what and how they respond. In focussing on how the teacher designs factual questions and how children respond to these questions it shows that question design can close down a student’s reply; or elicit a range of answers, from one word to extended more detailed responses. Understanding how the design of teachers’ questions can influence students’ responses has pedagogic implications and may support educators to make intentional decisions regarding their own questioning techniques.

Vote as you go : Blending interfaces for community engagement into the urban space

This paper presents a series of studies on situated interfaces for community engagement. Firstly, we identify five recurring design challenges as well as four common strategies used to overcome them. We then assess the effectiveness of these strategies through field studies with public polling interfaces. We developed two very different polling interfaces in the form of (1) a web application running on an iPad mounted on a stand, allowing one vote at a time, and (2) a playful full-body interaction application for a large urban screen allowing concurrent participation. We deployed both interfaces in an urban precinct with high pedestrian traffic and equipped with a large urban screen. Analysing discoverability and learnability of each scenario, we derive insights regarding effective ways of blending community engagement interfaces into the built environment, while attracting the attention of passers-by and communicating the results of civic participation.

Near infrared spectroscopy of marble from Chillagoe, Australia and in comparison with other selected natural carbonates

Marble from the Chillagoe deposits was extensively used in the construction of Australia’s parliament house. Near infrared (NIR) spectroscopy has been applied to study the quality of marble from the Chillagoe marble deposits and to distinguish between different types of marble in the Chillagoe deposits. A comparison of the NIR spectra of marble with dolomite and monohydrocalcite is made. The spectrum of the marble closely resembles that of monohydrocalcite and is different from that of dolomite. The infrared spectra of the minerals are characterised by OH and water stretching vibrations. Both the first and second fundamental overtones of these bands are observed in the NIR spectra. Marble is characterised by NIR bands at 4005, 4268 and 4340 cm–1, attributed to carbonate combination bands and overtones. Marble also shows NIR bands at 5005, 5106, 5234 and 5334 cm–1 assigned to water combination bands. Finally the NIR spectrum of the marble displays broad low-intensity features centred upon 6905 cm–1 attributed to the water first overtones. It appears feasible to identify marble through the use of NIR spectroscopy.

Projecting the new world city: The city as spectacle in an urban light festival

As a precursor to the 2014 G20 Leaders’ Summit held in Brisbane, Australia, the Queensland Government sponsored a program of G20 Cultural Celebrations, designed to showcase the Summit’s host city. The cultural program’s signature event was the Colour Me Brisbane festival, a two-week ‘citywide interactive light and projection installations’ festival that was originally slated to run from 24 October to 9 November, but which was extended due to popular demand to conclude with the G20 Summit itself on 16 November. The Colour Me Brisbane festival comprised a series projection displays that promoted visions of the city’s past, present, and future at landmark sites and iconic buildings throughout the city’s central business district and thus transformed key buildings into forms of media architecture. In some instances the media architecture installations were interactive, allowing the public to control aspects of the projections through a computer interface situated in front of the building; however, the majority of the installations were not interactive in this sense. The festival was supported by a website that included information regarding the different visual and interactive displays and links to social media to support public discussion regarding the festival (Queensland Government 2014). Festival-goers were also encouraged to follow a walking-tour map of the projection sites that would take them on a 2.5 kilometre walk from Brisbane’s cultural precinct, through the city centre, concluding at parliament house. In this paper, we investigate the Colour Me Brisbane festival and the broader G20 Cultural Celebrations as a form of strategic placemaking—designed, on the one hand, to promote Brisbane as a safe, open, and accessible city in line with the City Council’s plan to position Brisbane as a ‘New World City’ (Brisbane City Council 2014). On the other hand, it was deployed to counteract growing local concerns and tensions over the disruptive and politicised nature of the G20 Summit by engaging the public with the city prior to the heightened security and mobility restrictions of the Summit weekend. Harnessing perspectives from media architecture (Brynskov et al. 2013), urban imaginaries (Cinar & Bender 2007), and social media analysis, we take a critical approach to analysing the government-sponsored projections, which literally projected the city onto itself, and public responses to them via the official, and heavily promoted, social media hashtags (#colourmebrisbane and #g20cultural). Our critical framework extends the concepts of urban phantasmagoria and urban imaginaries into the emerging field of media architecture to scrutinise its potential for increased political and civic engagement. Walter Benjamin’s concept of phantasmagoria (Cohen 1989; Duarte, Firmino, & Crestani 2014) provides an understanding of urban space as spectacular projection, implicated in commodity and techno-culture. The concept of urban imaginaries (Cinar & Bender 2007; Kelley 2013)—that is, the ways in which citizens’ experiences of urban environments are transformed into symbolic representations through the use of imagination—similarly provides a useful framing device in thinking about the Colour Me Brisbane projections and their relation to the construction of place. Employing these critical frames enables us to examine the ways in which the installations open up the potential for multiple urban imaginaries—in the sense that they encourage civic engagement via a tangible and imaginative experience of urban space—while, at the same time, supporting a particular vision and way of experiencing the city, promoting a commodified, sanctioned form of urban imaginary. This paper aims to dissect the urban imaginaries intrinsic to the Colour Me Brisbane projections and to examine how those imaginaries were strategically deployed as place-making schemes that choreograph reflections about and engagement with the city.

Local Commons: A visual approach to collective city making through situated community engagement

Due to the numerous possibilities of voicing concerns and the flood of data we are exposed to, local issues are sometimes at risk of being overlooked. This study explores Local Commons, a design intervention in public space that combines situated digital and tangible media in order to engage communities in contributing and debating different perspectives on a given local issue. The intervention invited the community to submit images of their perspectives on the issue, which were displayed on a public screen. Via tangible buttons in front of the screen, community members then agree or disagree on the displayed perspectives, creating a space for deliberation. In a user study, we were specifically interested in testing three aspects of our intervention, which are discussed in this paper: The difference that situatedness, visual content, and tangible interaction can make to urban community engagement.

RNASeqBrowser: A genome browser for simultaneous visualization of raw strand specific RNAseq reads and UCSC genome browser custom tracks

Background Strand specific RNAseq data is now more common in RNAseq projects. Visualizing RNAseq data has become an important matter in Analysis of sequencing data. The most widely used visualization tool is the UCSC genome browser that introduced the custom track concept that enabled researchers to simultaneously visualize gene expression at a particular locus from multiple experiments. Our objective of the software tool is to provide friendly interface for visualization of RNAseq datasets. Results This paper introduces a visualization tool (RNASeqBrowser) that incorporates and extends the functionality of the UCSC genome browser. For example, RNASeqBrowser simultaneously displays read coverage, SNPs, InDels and raw read tracks with other BED and wiggle tracks -- all being dynamically built from the BAM file. Paired reads are also connected in the browser to enable easier identification of novel exon/intron borders and chimaeric transcripts. Strand specific RNAseq data is also supported by RNASeqBrowser that displays reads above (positive strand transcript) or below (negative strand transcripts) a central line. Finally, RNASeqBrowser was designed for ease of use for users with few bioinformatic skills, and incorporates the features of many genome browsers into one platform. Conclusions The features of RNASeqBrowser: (1) RNASeqBrowser integrates UCSC genome browser and NGS visualization tools such as IGV. It extends the functionality of the UCSC genome browser by adding several new types of tracks to show NGS data such as individual raw reads, SNPs and InDels. (2) RNASeqBrowser can dynamically generate RNA secondary structure. It is useful for identifying non-coding RNA such as miRNA. (3) Overlaying NGS wiggle data is helpful in displaying differential expression and is simple to implement in RNASeqBrowser. (4) NGS data accumulates a lot of raw reads. Thus, RNASeqBrowser collapses exact duplicate reads to reduce visualization space. Normal PC’s can show many windows of NGS individual raw reads without much delay. (5) Multiple popup windows of individual raw reads provide users with more viewing space. This avoids existing approaches (such as IGV) which squeeze all raw reads into one window. This will be helpful for visualizing multiple datasets simultaneously. RNASeqBrowser and its manual are freely available at http://www.australianprostatecentre.org/research/software/rnaseqbrowser webcite or http://sourceforge.net/projects/rnaseqbrowser/ webcite

ITO-free top emitting organic light emitting diodes with enhanced light out-coupling

Bottom emitting organic light emitting diodes (OLEDs) can suffer from lower external quantum efficiencies (EQE) due to inefficient out-coupling of the generated light. Herein, it is demonstrated that the current efficiency and EQE of red, yellow, and blue fluorescent single layer polymer OLEDs is significantly enhanced when a MoOx(5 nm)/Ag(10 nm)/MoOx(40 nm) stack is used as the transparent anode in a top emitting OLED structure. A maximum current efficiency and EQE of 21.2 cd/A and 6.7%, respectively, was achieved for a yellow OLED, while a blue OLED achieved a maximum of 16.5 cd/A and 10.1%, respectively. The increase in light out-coupling from the top-emitting OLEDs led to increase in efficiency by a factor of up to 2.2 relative to the optimised bottom emitting devices, which is the best out-coupling reported using solution processed polymers in a simple architecture and a significant step forward for their use in large area lighting and displays.

Media futures: A review essay on'The Future of Reputation','TV Futures', and'The Future of the Internet and How to Stop It'

In his 1987 book, The Media Lab: Inventing the Future at MIT, Stewart Brand provides an insight into the visions of the future of the media in the 1970s and 1980s. 1 He notes that Nicolas Negroponte made a compelling case for the foundation of a media laboratory at MIT with diagrams detailing the convergence of three sectors of the media—the broadcast and motion picture industry; the print and publishing industry; and the computer industry. Stewart Brand commented: ‘If Negroponte was right and communications technologies really are converging, you would look for signs that technological homogenisation was dissolving old boundaries out of existence, and you would expect an explosion of new media where those boundaries used to be’. Two decades later, technology developers, media analysts and lawyers have become excited about the latest phase of media convergence. In 2006, the faddish Time Magazine heralded the arrival of various Web 2.0 social networking services: You can learn more about how Americans live just by looking at the backgrounds of YouTube videos—those rumpled bedrooms and toy‐strewn basement rec rooms—than you could from 1,000 hours of network television. And we didn’t just watch, we also worked. Like crazy. We made Facebook profiles and Second Life avatars and reviewed books at Amazon and recorded podcasts. We blogged about our candidates losing and wrote songs about getting dumped. We camcordered bombing runs and built open‐source software. America loves its solitary geniuses—its Einsteins, its Edisons, its Jobses—but those lonely dreamers may have to learn to play with others. Car companies are running open design contests. Reuters is carrying blog postings alongside its regular news feed. Microsoft is working overtime to fend off user‐created Linux. We’re looking at an explosion of productivity and innovation, and it’s just getting started, as millions of minds that would otherwise have drowned in obscurity get backhauled into the global intellectual economy. The magazine announced that Time’s Person of the Year was ‘You’, the everyman and everywoman consumer ‘for seizing the reins of the global media, for founding and framing the new digital democracy, for working for nothing and beating the pros at their own game’. This review essay considers three recent books, which have explored the legal dimensions of new media. In contrast to the unbridled exuberance of Time Magazine, this series of legal works displays an anxious trepidation about the legal ramifications associated with the rise of social networking services. In his tour de force, The Future of Reputation: Gossip, Rumor, and Privacy on the Internet, Daniel Solove considers the implications of social networking services, such as Facebook and YouTube, for the legal protection of reputation under privacy law and defamation law. Andrew Kenyon’s edited collection, TV Futures: Digital Television Policy in Australia, explores the intersection between media law and copyright law in the regulation of digital television and Internet videos. In The Future of the Internet and How to Stop It, Jonathan Zittrain explores the impact of ‘generative’ technologies and ‘tethered applications’—considering everything from the Apple Mac and the iPhone to the One Laptop per Child programme.

Commercial friendships between gay sales associates and straight female customers in luxury settings: A proposed theoretical framework

This study investigates friendships between gay sales associates and heterosexual female customers in luxury retail settings. By employing grounded theory methodology, the study integrates theories and findings from diverse literature streams into an original conceptual framework to illustrate the resources gay sales associates and straight female customers receive from and provide to each other during retail exchanges. The study explains why gay male–straight female friendships are uniquely suited for luxury consumption settings. Female customers characterize their friendships with gay sales associates as providing honesty, security, trust, and comfort, which stems from the absence of sexual interest and a lack of inter-female competition. Gay sales associates receive acceptance for who they are and for their displays of unconventional masculinity in retail settings. They also obtain a temporary rite from their female customers, a so-called mandate of privacy, which permits both parties to ignore the bounds of modesty and accept a degree of intimacy. Such intimacy facilitates transactions that require both personalization and customer–employee closeness, such as the selling of high-end apparel, accessories, and jewelry.

Digital design interventions for creating new presentations of self in public urban places

During everyday urban life, people spend time in public urban places waiting for specific events to occur. During these times, people sometimes tend to engage with their information and communication technology (ICT) devices in a way that shuts off interactions with collocated people. These devices could also be used to better connect with the urban space and collocated people within. This chapter presents and discusses the impact of three design interventions on the urban user experience enabling collocated people to share lightweight, non-privacy-sensitive data in the urban space. We investigate and discuss the impact on the urban experience under the notions of people, place, and technology with an emphasis on how the sharing of non-privacy-sensitive data can positively transform anonymous public urban places in various ways through anonymous digital augmentations.