Expression and purification of soluble recombinant full length HIV-1 Pr55(Gag) protein in Escherichia coli

The HIV-1 Gag precursor protein, Pr55(Gag), is a multi-domain polyprotein that drives HIV-1 assembly. The morphological features of HIV-1 suggested Pr55(Gag) assumes a variety of different conformations during virion assembly and maturation, yet structural determination of HIV-1 Pr55(Gag) has not been possible due to an inability to express and to isolate large amounts of full-length recombinant Pr55(Gag) for biophysical and biochemical analyses. This challenge is further complicated by HIV-1 Gag's natural propensity to multimerize for the formation of viral particle (with ∼2500 Gag molecules per virion), and this has led Pr55(Gag) to aggregate and be expressed as inclusion bodies in a number of in vitro protein expression systems. This study reported the production of a recombinant form of HIV-1 Pr55(Gag) using a bacterial heterologous expression system. Recombinant HIV-1 Pr55(Gag) was expressed with a C-terminal His×6 tag, and purified using a combination of immobilized metal affinity chromatography and size exclusion chromatography. This procedure resulted in the production of milligram quantities of high purity HIV-1 Pr55(Gag) that has a mobility that resembles a trimer in solution using size exclusion chromatography analysis. The high quantity and purity of the full length HIV Gag will be suitable for structural and functional studies to further understand the process of viral assembly, maturation and the development of inhibitors to interfere with the process.

Molecular and immunological analysis of hen's egg yolk allergens with a focus on YGP42 (Gal d 6)

Allergy to hen's (Gallus domesticus) egg white is one of the most common forms of food allergy. Allergy to hen's yolk also exists however, to a lesser extent when compared to egg white allergy. Two minor allergens from the hen's egg yolk known as α-livetin (Gal d 5) and YGP42 (Gal d 6) were discovered recently. In this study, we investigated whether sensitization to egg white is associated with reactivity to egg yolk as well. Sera obtained from 25 patients with allergy to egg white were tested for specific IgE binding for egg yolk proteins through western immunoblotting. 36% of patients were found with true IgE-sensitization against egg yolk proteins. It was found that most of the IgE reactive yolk proteins were fragments of major precursor proteins of hen; vitellogenin-1 (VTG-1), vitellogenin-2 (VTG-2) and apolipoprotein B (Apo B). The egg yolk allergen Gal d 6 is the C-terminal part of VTG-1 and was found to be allergenic in significant percentage of egg white allergy patients. These results highlight the significance of Gal d 6 as an important allergen of egg yolk. Therefore, the secondary aim of this study involved developing a recombinant version of YGP42 in an Escherichia coli expression system. Recombinant Gal d 6 (rGal d6) was expressed as a fusion peptide with a 6 × His tag and purified using metal chelating resin. The inhibition ELISA results showed that rYGP42 was IgE reactive and was able to inhibit IgE binding to crude egg yolk (CEY) by up to 30%. Traditionally, it was thought that allergy to egg yolk occurred independently from sensitization to egg white. This study underlies the importance of concomitant sensitization to egg yolk proteins in patients allergic to egg white. Evidence reported in this study strongly suggests that egg yolk has potentially undiscovered allergens and therefore warrants further investigation. Furthermore, IgE reactive Gal d 6 presented in this study has the potential to be used in diagnosis and immunotherapy to treat egg allergy.

Inclusion in political and public life: the experiences of people with intellectual disability on government disability advisory bodies in Australia

BACKGROUND: Civil and political participation lies at the core of citizenship. Increasingly, people with intellectual disability are members of disability advisory bodies. This study investigated the political orientations of advisory body members with intellectual disability, their participatory experiences, and the types of support they received. METHOD: The 9 people with intellectual disability who in 2005 were members of disability advisory bodies at a state, national, and Victorian local government level were interviewed, together with 12 other members or secretariat staff of these bodies. Observations were also conducted of advisory body meetings. RESULTS: The political perspective of members with intellectual disability varied, but all had a background in self-advocacy. They found the work hard but rewarding and encountered both practical and intangible obstacles to participation. Members received varying types of practical support, but a supportive collegial milieu was characteristic among members who felt most confident about their participation. CONCLUSIONS: The milieu, structures, and processes of advisory bodies must all be adjusted to accommodate people with intellectual disability if they are to participate meaningfully.

Inclusion and accountability: communications research with consumers with disabilities

Over the past two decades there has been an increasing focus on doing research with and for consumers rather than on consumers. Research that is collaborative and inclusive has been called for by people with disabilities. People with disabilities are a group of telecommunications consumers who are disadvantaged because of social barriers and access issues with equipment, services and information. In this paper, the benefits and challenges of collaborative (e.g. participatory, participatory action, emancipatory) research with these consumers and the processes that can facilitate inclusive and accountable research outcomes are addressed. Throughout the paper, four questions will be explored: Who determines which research issues are explored and the methods that are used? What is the level of consumer involvement in the research process? How can academics and government ensure that consumer-focused research is inclusive and participatory? How, and to whom, should research results be made available?

In the first part of the paper, the perspectives on consumer research by people with disabilities are presented. Respect for and appreciation of consumers' views underpins the philosophical stance needed by researchers and funding bodies before serious engagement in empowering, person-centred research can be successful.

In the second part of the paper, there is a focus on defining the features of collaborative research approaches. Participatory research, participatory action research, and emancipatory research are variations that incorporate differing levels of consumer involvement. Each of these, however, embrace shifts in philosophy and methods away from more traditional 'scientific' research. Such changes result in a greater sense of inclusion and project ownership by consumers.

The third part of the paper addresses some of the issues associated with collaborative research for consumers with disabilities, for academic researchers, and for government funding bodies. Differing understandings of the social nature of disablement influence attitudes toward consumer involvement and are demonstrated in the choice of research questions, project aims, and the methodologies used. Differing agendas are reflected in reactions to the traditional project requirements and processes of government, the preferred research methodologies of researchers, and the perceived accountability of researchers or government for the dissemination and sharing of research results. The challenges to successful engagement with consumers in collaborative research have been identified in the literature and a number of strategies recommended (Barnes, 2003; Clear, 1999; Mercer, 2002; Oliver, 1992, 1997; Zarb, 1997). Application of inclusive strategies by researchers and funding bodies is essential for positive change and inclusive, empowering outcomes.

The paper concludes with a case study of a DCITA-funded research project. The project is evaluated using participatory/emancipatory research criteria modified from Zarb (1992) and addresses consumer, researcher, and funding body participation. The four questions used in the discussion of the paper are applied to this particular project. In addition, the benefits and learnings from the project will be compared with the outcomes desired by consumers who espouse collaborative research approaches.

Performance and narrative, bodies and movement in the construction of places and objects, spaces and knowledges: The case of the Maltese megaliths

The article explores the ways knowledge and space are co-produced performatively through bodily movement in an examination of the Maltese megaliths the first complex stone structures in the world. It is argued that knowledge is best seen as spatialized narratives of human actions and objects as materialized forms of those spatial narratives. Rewriting our social and historical narratives so that the performativity of place, space and knowledge is restored opens new possibilities for rethinking the social and material order.

De Chirico and Walker in light of Nietzsche and Guattari: the enigma of partial bodies in illogical spaces

If one concedes that the Freudian unconscious is inseparable from a society attached to its past, for example, its phallocentric traditions, Guattari’s alternative model dealing with "the production of subjectivity" offers a new perspective (1995: 11). From this vantage point, it is possible to map the way "every individual and social group" models the creation of subjectivity, a subjectivity "composed of cognitive references as well as mythical, ritual and symptomatological references" (1995: 11).

Queer trepidations and the art of inclusion

Contemporary research and pedagogy related to sexualities and schooling in Australia, Aotearoa1/New Zealand and the United States often focuses on ways to alleviate homophobia and heterosexism in the hope of creating schools that are more inclusive of lesbian and gay (and very rarely bisexual, transgender and intersex2) (LGBTI) teachers and students. Within this paradigm, the notion of what comprises sexualities is often taken as given. Alternatively, researchers and educators may invoke essentialising narratives in order to make arguments for the inclusion of students and teachers who adopt LGBTI identifications. Drawing on a theoretical framework influenced by the work of Deborah Britzman3 and other queer theorists within and outside education this article interrogates these strategies of inclusion. In particular, I focus on research methodologies and pedagogies related to sexualities and schooling devised in the name of inclusion of young people who identify as lesbian, gay, bisexual or transgender (LGBT)4 in secondary educational contexts. This analysis, which is based on my doctoral studies, commences with a consideration of queer theories and the art of inclusion. Subsequent to this I analyse pedagogies of inclusion and methodologies of inclusion, and, their nexus with queer theories.

Conspiring to run: women, their bodies and athletics training

Women's athletics commenced about 60 years after the start of the men's sport. Women's involvement in athletics was held back by the medical and general views that this was a strenuous sport requiring a level of exertion beyond the biological capabilities of female bodies. Their difficult initiation into athletics occurred under male gaze; they encountered opposition from the public, the medical profession and from the male-controlled athletics organizations. A serious participation in athletics requires significant exertion and dedicated training. While the prevailing view was that moderate physical exercise without strain enhanced women's health, the exertion required for athletics was deemed to be potentially dangerous. Within essentialist views of gender, women's involvement in athletics was thought to have implications for their nurturing and domestic roles. When the pioneer women athletes tried to excel, they were said to be straining themselves and their participation in the sport was brought into question. By using theoretical insights drawn mostly from Foucault, training manuals from the early decades of women's athletics and material from interviews with some of the first English female athletes are examined to investigate the attitudes of both genders to women in athletics and to analyse how they circumvented the potential veto of their sport by men.

Bad, mad and sad : developing a methodology of inclusion and a pedagogy for researching students with intellectual disabilities

A critical question for me as a teacher/researcher in the field of inclusive education is how to reposition children with moderate and severe intellectual disabilities as participants rather than subjects in the debate. In this paper, I develop a methodology of inclusion that comprises an ethics of consent and a pedagogy for research participation that is an opportunity not only to teach, but also to create a new discursive space for six children to speak. The discussion explores a range of methodological and interpretive strategies for including children with significant intellectual disabilities in research: issues of informed consent, the negotiation of power relations and the ways in which this innovative pedagogy can be empowering beyond the research situation. The use of this methodology has provocative implications concerning what might be learned about forging a link between the struggle for change and educational policy/practice if other researchers worked towards creating spaces for these most marginalized children to speak.

Category politics: deaf students' inclusion in a 'hearing university'

This article investigates the way in which deaf tertiary students' identity is constructed within the university - an overwhelmingly 'hearing' institution. It is a descriptive and analytical account of the experiences of two deaf teacher education students as they reflect on their progress and experiences in higher education. Data have been analysed within an interpretive framework of category politics and the construction of difference. The study found that providing the same access to the same information in the same form did little to address the discursive marginality of these students.

Working with 'diverse bodies, diverse identities': an approach to professional education about 'diversity'

The complexity and diversity of populations in contemporary Western societies is becoming a significant public policy issue. The concept of 'diversity' has come to represent cultural, ethnic, racial and religious differences between the 'dominant group' and immigrant and indigenous populations. 'Diversity training' is amongst many strategies being implemented to address social and economic objectives in complex societies. This paper discusses and critically evaluates a professional education programme, 'Diverse Bodies, Diverse Identities', that is offered to human service practitioners and social work students in Victoria, Australia. It is concluded that a range of approaches is needed to address 'diversity' in contemporary societies.

Gatekeeping or redressing social exclusion : expectations on social work educators in relation to incarcerated students

Until recently, the author was in Scotland, where professional registration in social work extends to students and involves criminal record checks prior to acceptance into a course of study. She is now teaching at Deakin University in Australia, which places a high priority on making higher education available to persons and groups who have traditionally been excluded, both through the provision of courses through off campus (distance education) study mode and an innovative and culturally sensitive mode of provision for indigenous Australians. One result of our attempts to redress social exclusion is that, on occasion, we discover that some of our students are incarcerated. There are important logistical issues which may emerge as a consequence of accepting prisoners into a program of social work education. However, it would seem that the inclusion of prisoners is symbolic of a fundamental difference in philosophy with programmes of social work education in countries where there is a strong expectation that social work educators act as gatekeepers to the profession, especially in respect of students with criminal convictions. This in tum reflects an expectation among social work educators in Australia that it may be more appropriate for professional associations or registration bodies to determine whether or not a graduate with a criminal record is suitable for employment as a professional social worker. In some settings, a prior criminal record is not a barrier to being an effective service provider, as well as international differences in understandings of the social work role and employment
destinations of social work graduates.