Measurement of Epstein-Barr virus DNA load using a novel quantification standard containing two EBV DNA targets and SYBR Green I dye

BACKGROUND Reactivation of Epstein-Barr virus (EBV) infection may cause serious, life-threatening complications in immunocompromised individuals. EBV DNA is often detected in EBV-associated disease states, with viral load believed to be a reflection of virus activity. Two separate real-time quantitative polymerase chain reaction (QPCR) assays using SYBR Green I dye and a single quantification standard containing two EBV genes, Epstein-Barr nuclear antigen-1 (EBNA-1) and BamHI fragment H rightward open reading frame-1 (BHRF-1), were developed to detect and measure absolute EBV DNA load in patients with various EBV-associated diseases. EBV DNA loads and viral capsid antigen (VCA) IgG antibody titres were also quantified on a population sample. RESULTS EBV DNA was measurable in ethylenediaminetetraacetic acid (EDTA) whole blood, peripheral blood mononuclear cells (PBMCs), plasma and cerebrospinal fluid (CSF) samples. EBV DNA loads were detectable from 8.0 × 10² to 1.3 × 10⁸ copies/ml in post-transplant lymphoproliferative disease (n = 5), 1.5 × 10³ to 2.0 × 10⁵ copies/ml in infectious mononucleosis (n = 7), 7.5 × 10⁴ to 1.1 × 10⁵ copies/ml in EBV-associated haemophagocytic syndrome (n = 1), 2.0 × 10² to 5.6 × 10³ copies/ml in HIV-infected patients (n = 12), and 2.0 × 10² to 9.1 × 10⁴ copies/ml in the population sample (n = 218). EBNA-1 and BHRF-1 DNA were detected in 11.0% and 21.6% of the population sample respectively. There was a modest correlation between VCA IgG antibody titre and BHRF-1 DNA load (rho = 0.13, p = 0.05) but not EBNA-1 DNA load (rho = 0.11, p = 0.11). CONCLUSION Two sensitive and specific real-time PCR assays using SYBR Green I dye and a single quantification standard containing two EBV DNA targets, were developed for the detection and measurement of EBV DNA load in a variety of clinical samples. These assays have application in the investigation of EBV-related illnesses in immunocompromised individuals.The Ausimmune Study is funded by the National Multiple Sclerosis Society of the USA, the National Health & Medical Research Council (Project Grant 316901) and Multiple Sclerosis Research Australia

Deepening our Understanding of Quality in Australia (DUQuA): a study protocol for a nationwide, multilevel analysis of relationships between hospital quality management systems and patient factors.

INTRODUCTION: Despite the growing body of research on quality and safety in healthcare, there is little evidence of the association between the way hospitals are organised for quality and patient factors, limiting our understanding of how to effect large-scale change. The 'Deepening our Understanding of Quality in Australia' (DUQuA) study aims to measure and examine relationships between (1) organisation and department-level quality management systems (QMS), clinician leadership and culture, and (2) clinical treatment processes, clinical outcomes and patient-reported perceptions of care within Australian hospitals. METHODS AND ANALYSIS: The DUQuA project is a national, multilevel, cross-sectional study with data collection at organisation (hospital), department, professional and patient levels. Sample size calculations indicate a minimum of 43 hospitals are required to adequately power the study. To allow for rejection and attrition, 70 hospitals across all Australian jurisdictions that meet the inclusion criteria will be invited to participate. Participants will consist of hospital quality management professionals; clinicians; and patients with stroke, acute myocardial infarction and hip fracture. Organisation and department-level QMS, clinician leadership and culture, patient perceptions of safety, clinical treatment processes, and patient outcomes will be assessed using validated, evidence-based or consensus-based measurement tools. Data analysis will consist of simple correlations, linear and logistic regression and multilevel modelling. Multilevel modelling methods will enable identification of the amount of variation in outcomes attributed to the hospital and department levels, and the factors contributing to this variation. ETHICS AND DISSEMINATION: Ethical approval has been obtained. Results will be disseminated to individual hospitals in de-identified national and international benchmarking reports with data-driven recommendations. This ground-breaking national study has the potential to influence decision-making on the implementation of quality and safety systems and processes in Australian and international hospitals

Antigen-presenting genes and genomic copy number variations in the Tasmanian devil MHC

BACKGROUND The Tasmanian devil (Sarcophilus harrisii) is currently under threat of extinction due to an unusual fatal contagious cancer called Devil Facial Tumour Disease (DFTD). DFTD is caused by a clonal tumour cell line that is transmitted between unrelated individuals as an allograft without triggering immune rejection due to low levels of Major Histocompatibility Complex (MHC) diversity in Tasmanian devils. RESULTS Here we report the characterization of the genomic regions encompassing MHC Class I and Class II genes in the Tasmanian devil. Four genomic regions approximately 960 kb in length were assembled and annotated using BAC contigs and physically mapped to devil Chromosome 4q. 34 genes and pseudogenes were identified, including five Class I and four Class II loci. Interestingly, when two haplotypes from two individuals were compared, three genomic copy number variants with sizes ranging from 1.6 to 17 kb were observed within the classical Class I gene region. One deletion is particularly important as it turns a Class Ia gene into a pseudogene in one of the haplotypes. This deletion explains the previously observed variation in the Class I allelic number between individuals. The frequency of this deletion is highest in the northwestern devil population and lowest in southeastern areas. CONCLUSIONS The third sequenced marsupial MHC provides insights into the evolution of this dynamic genomic region among the diverse marsupial species. The two sequenced devil MHC haplotypes revealed three copy number variations that are likely to significantly affect immune response and suggest that future work should focus on the role of copy number variations in disease susceptibility in this species.This work was funded by an ARC Future Fellowship to KB (FT0992212), the Eric Guiler fund and the Tasmanian Department of Primary Industries, Parks, Water and the Environment. YC was supported by an Endeavour International Postgraduate Research Scholarship, KM by an Australian Postgraduate Award and an ARC Linkage Grant

Telomere dynamics and homeostasis in a transmissible cancer

Devil Facial Tumour Disease (DFTD) is a unique clonal cancer that threatens the world\u27s largest carnivorous marsupial, the Tasmanian devil (Sarcophilus harrisii) with extinction. This transmissible cancer is passed between individual devils by cell implantation during social interactions. The tumour arose in a Schwann cell of a single devil over 15 years ago and since then has expanded clonally, without showing signs of replicative senescence; in stark contrast to a somatic cell that displays a finite capacity for replication, known as the “Hayflick limit”

Towards a practice turn in critical management studies : manifesting a dream through NGO engagement with corporate social responsibility

This research takes an anamorphic gaze on how to influence the development of social responsible business practice by looking at how non-government organisations (NGOs) collaborate with corporations. The study proposed that a strategically motivated type of NGO engagement can uncover new attitudes to the practice of corporate social responsibility (CSR), and offer insights into how the application of Critical Management Studies (CMS) can change from a relatively static analytical exercise to a more dynamic critical form of enterprise practice. The study challenges the traditional business-centric understanding of CSR. Particularly, it shifts the focus. CSR is often thought of in business terms as a type of practice where corporations have a choice about how they might contribute to society. This is sometimes framed as a corporate duty to contribute to the economy, obey laws, be ethical and philanthropically contribute to society (Carroll 1991). This business-centric perspective of CSR is almost exclusively focused on the corporation and its own imperatives and inclinations to unilaterally address its social responsibilities. However, the more transformative perspective of CSR adopted in this research allows other sectors of society to contribute to a corporation’s socially responsible conduct. Motivated by their own interests, these sectors can be understood to have the capacity to exert a level of influence over corporations. This approach draws on a CSR tradition that Utting (2002) refers to as the ‘regulatory frame of CSR’. The regulatory perspective enables policy based entities, private firms, and civil society organisations to monitor – to “regulate” – corporate activity, to intervene when appropriate and to influence corporations in how they exercise their social responsibilities. This is depicted in Utting’s (2002) three dimensions of CSR: ‘command and control’, ‘corporate self-regulation’ and ‘stakeholder co-regulation’. The dimension of ‘stakeholder co-regulation’ forms the frame of this research. This regulatory perspective of CSR can be understood to have similar principles to Critical Management Studies (CMS). CMS is a research construct that challenges those management activities and practices that appear to subjugate human needs and desires to the institutional profit-seeking tendencies of corporations (Fournier & Grey 2000). Both CMS and the regulatory practice of CSR take a problem-centred focus on the question of corporation behaviour in society; and both have an agenda for change. These similarities draw a connection between the two theoretical frames, and this link provides a channel by which the regulatory frame of CSR could be imagined and understood in the context of CMS. In this regulatory frame of CSR, the research has focused specifically on how NGOs participated in ‘co-regulating’ corporations to work together on developing socially responsible business practice. In particular, the study concentrated on how NGOs use collaborative processes to do this. I developed a multi-phased action research framework to provide a scaffold for the collaboration between the NGOs and corporations, and to mitigate some of the risks associated with the NGOs being co-opted to the corporate perspectives. The framework included a synthesis of action learning and appreciative inquiry approaches. Three NGOs from Australia’s social services sector were recruited for the study. Each adapted the Action Research framework to suit their own needs and objectives for their engagement with the private sector. One of the NGOs used action learning and appreciative inquiry processes to support corporations to participate in community projects. Another chose to use appreciative inquiry for the same purpose. In contrast, the third NGO used action learning processes to resolve some of its internal challenges to corporate engagement. The NGOs were found to have drawn on action learning and appreciative inquiry as separate and distinct processes, but in interconnected and complementary ways. The research revealed that the NGOs were not seeking to ‘co-regulate’ corporate behaviour in the Utting (2002) tradition: they were not looking to monitor corporate activity and they were not looking to influence it for the purposes of improving the corporation’s social performance. Instead, the NGOs had sought to design and direct the manner in which corporations could participate in addressing community-based aims and objectives. This pointed to the existence of an additional frame of CSR; one that moved beyond the ‘regulatory’ frames, to adopt a ‘developmental agenda’ that was more visionary. This type of CSR offered scope to extend Utting’s (2002) regulatory-based framework to include a developmental dimension that I refer to as: ‘stakeholder-directive co-development’. The developmental frame denotes instances where stakeholders direct corporations about how they can participate in achieving stakeholder objectives. This study makes a contribution to knowledge by uncovering stakeholder-directive co-development as a new frame of CSR. This new frame offers an opportunity to challenge the dominant, problem-centred perspective of corporate activity adopted in CSR and CMS. The implications of this indicate that the ‘problem-centred heart’ of these frames may not advance the kind of social change dreamt of by those who pursue CSR and CMS. Through this dissertation I propose that the inclusion of a stakeholder-directive co-development agenda could confer the missing ‘link’ needed to transform those social dreams into a reality