DNA technology for the detection of common genetic variants that predispose to thrombophilia

With the identification of common single locus point mutations as risk factors for thrombophilia, many DNA testing methodologies have been described for detecting these variations. Traditionally, functional or immunological testing methods have been used to investigate quantitative anticoagulant deficiencies. However, with the emergence of the genetic variations, factor V Leiden, prothrombin 20210 and, to a lesser extent, the methylene tetrahydrofolate reductase (MTHFR677) and factor V HR2 haplotype, traditional testing methodologies have proved to be less useful and instead DNA technology is more commonly employed in diagnostics. This review considers many of the DNA techniques that have proved to be useful in the detection of common genetic variants that predispose to thrombophilia. Techniques involving gel analysis are used to detect the presence or absence of restriction sites, electrophoretic mobility shifts, as in single strand conformation polymorphism or denaturing gradient gel electrophoresis, and product formation in allele-specific amplification. Such techniques may be sensitive, but are unwielding and often need to be validated objectively. In order to overcome some of the limitations of gel analysis, especially when dealing with larger sample numbers, many alternative detection formats, such as closed tube systems, microplates and microarrays (minisequencing, real-time polymerase chain reaction, and oligonucleotide ligation assays) have been developed. In addition, many of the emerging technologies take advantage of colourimetric or fluorescence detection (including energy transfer) that allows qualitative and quantitative interpretation of results. With the large variety of DNA technologies available, the choice of methodology will depend on several factors including cost and the need for speed, simplicity and robustness. © 2000 Lippincott Williams & Wilkins.

Multiple analysis of three common genetic alterations associated with thrombophilia

We have previously reported the use of a novel mini-sequencing protocol for detection of the factor V Leiden variant, the first nucleotide change (FNC) technology. This technology is based on a single nucleotide extension of a primer, which is hybridized immediately adjacent to the site of mutation. The extended nucleotide that carries a reporter molecule (fluorescein) has the power to discriminate the genotype at the site of mutation. More recently, the prothrombin 20210 and thermolabile methylene tetrahydrofolate reductase (MTHFR) 677 variants have been identified as possible risk factors associated with thrombophilia. This study describes the use of the FNC technology in a combined assay to detect factor V, prothrombin and MTHFR variants in a population of Australian blood donors, and describes the objective numerical methodology used to determine genotype cut-off values for each genetic variation. Using FNC to test 500 normal blood donors, the incidence of Factor V Leiden was 3.6% (all heterozygous), that of prothrombin 20210 was 2.8% (all heterozygous) and that of MTHFR was 10% (homozygous). The combined FNC technology offers a simple, rapid, automatable DNA-based test for the detection of these three important mutations that are associated with familial thrombophilia. (C) 2000 Lippincott Williams and Wilkins.

Use of first nucleotide change technology to determine the frequency of factor V Leiden in a population of Australian blood donors

Activated protein C resistance (APCR), the most common risk factor for venous thrombosis, is the result of a G to A base substitution at nucleotide 1691 (R506Q) in the factor V gene. Current techniques to detect the factor V Leiden mutation, such as determination of restriction length polymorphisms, do not have the capacity to screen large numbers of samples in a rapid, cost- effective test. The aim of this study was to apply the first nucleotide change (FNC) technology, to the detection of the factor V Leiden mutation. After preliminary amplification of genomic DNA by polymerase chain reaction (PCR), an allele-specific primer was hybridised to the PCR product and extended using fluorescent terminating dideoxynucleotides which were detected by colorimetric assay. Using this ELISA-based assay, the prevalence of the factor V Leiden mutation was determined in an Australian blood donor population (n = 500). A total of 18 heterozygotes were identified (3.6%) and all of these were confirmed with conventional MnlI restriction digest. No homozygotes for the variant allele were detected. We conclude from this study that the frequency of 3.6% is compatible with others published for Caucasian populations. In addition, the FNC technology shows promise as the basis for a rapid, automated DNA based test for factor V Leiden.

Building support for learning within a Doctor of Education programme

Professional doctorates were introduced in the 1990s for practitioners to research ‘real-world’ problems relevant to their respective workplace communities and contexts. An array of difficulties faces professional doctoral students as they transition from professionals to practitioner researchers. This study sought to understand the learning journey of a cohort of students at an Australian university and to assess whether the cohort approach provided the necessary support for students to reach their scholarly destinations. Throughout the first 18 months of the programme, focus group interviews and surveys were conducted to gauge students’ experiences and to evaluate developments for support within the programme. Utilising a socio-cultural perspective helped identify and explain the importance of shared practice in fostering learning, the development of academic and researcher identities, and the role of communities of practice. Challenges of managing time and overcoming the professional and academe divide were facilitated by the evolving developments of the programme.

Investigating child participation in the everyday talk of teacher and children in a preparatory year

In early years research, policy and education, a democratic perspective that positions children as participants and citizens is increasingly emphasized. These ideas take seriously listening to children’s opinions and respecting children’s influence over their everyday affairs. While much political and social investment has been paid to the inclusion of participatory approaches little has been reported on the practical achievement of such an approach in the day to day of early childhood education within school settings. This paper investigates talk and interaction in the everyday activities of a teacher and children in an Australian preparatory class (for children age 4-6 years) to see how ideas of child participation are experienced. We use an interactional analytic approach to demonstrate how participatory methods are employed in practical ways to manage routine interactions. Analysis shows that whilst the teacher seeks the children’s opinion and involves them in decision-making, child participation is at times constrained by the context and institutional categories of “teacher” and “student” that are jointly produced in their talk. The paper highlights tensions that arise for teachers as they balance a pedagogical intent of “teaching” and the associated institutional expectations, with efforts to engage children in decision-making. Recommendations include adopting a variety of conversational styles when engaging with children; consideration of temporal concerns and the need to acknowledge the culture of the school.

“What’s going on here?” The pedagogy of a data analysis session

Data analysis sessions are a common feature of discourse analytic communities, often involving participants with varying levels of expertise to those with significant expertise. Learning how to do data analysis and working with transcripts, however, are often new experiences for doctoral candidates within the social sciences. While many guides to doctoral education focus on procedures associated with data analysis (Heath, Hindmarsh, & Luff, 2010; McHoul & Rapley, 2001; Silverman, 2011; Wetherall, Taylor, & Yates, 2001), the in situ practices of doing data analysis are relatively undocumented. This chapter has been collaboratively written by members of a special interest research group, the Transcript Analysis Group (TAG), who meet regularly to examine transcripts representing audio- and video-recorded interactional data. Here, we investigate our own actual interactional practices and participation in this group where each member is both analyst and participant. We particularly focus on the pedagogic practices enacted in the group through investigating how members engage in the scholarly practice of data analysis. A key feature of talk within the data sessions is that members work collaboratively to identify and discuss ‘noticings’ from the audio-recorded and transcribed talk being examined, produce candidate analytic observations based on these discussions, and evaluate these observations. Our investigation of how talk constructs social practices in these sessions shows that participants move fluidly between actions that demonstrate pedagogic practices and expertise. Within any one session, members can display their expertise as analysts and, at the same time, display that they have gained an understanding that they did not have before. We take an ethnomethodological position that asks, ‘what’s going on here?’ in the data analysis session. By observing the in situ practices in fine-grained detail, we show how members participate in the data analysis sessions and make sense of a transcript.

Book Review: "Rethinking Learning in Early Childhood Education" written by Nicola Yelland, Libby Lee, Maureen O’Rourke & Cathie Harrison

There is a growing body of work that responds to the impact of the rapid uptake of information and communication technology (ICT) on education (Buckingham, 2003; Cheung, 2003; Cuban, 2003; Leung, 2003; Prensky, 2005; Green & Hannon, 2007; Brooks-Gunn & Donahue, 2008; Lyman et al, 2008). Mostly, this work has been positioned in the context of upper-primary or secondary classrooms. More recently, there has been a growing call for research about the impact of ICT on the early years or in early childhood contexts. This text initiates a response to that call. The authors concur that today’s children are a generation who create, learn, work, play and communicate very differently from their parents and teachers (Buckingham, 2003), and that classroom activity needs to reflect this difference.

Occupational health nurses’ roles, credentials, and continuing education in Ontario, Canada

The role of the occupational health nurse is broad and includes health care provider, manager/coordinator, educator/advisor, and case manager and consultant, depending on the type of industry and the country in which the nurse practices. Regardless of the type of role, the occupational health nurse must participate in continuing nursing education (CNE) activities. This study describes the roles, credentials, and number of CNE activities undertaken by occupational health nurses working in Ontario, Canada. Using a non-experimental descriptive design, a questionnaire was mailed to all practicing occupational health nurses who are members (n = 900) of a local nursing association. Three hundred fifty-four questionnaires were returned. Nurses reported a variety of roles in the following categories: case management, health promotion, policy development, infection control/travel health, ergonomics, education, research, health and safety, direct care, consultation, disaster preparedness, and industrial hygiene. Sixty-five percent of nurses held an occupational health nurse credential, and 19% of nurses attended more than 100 hours of CNE annually. Occupational health nurses have multiple workplace roles. Many attend CNE activities and they often prepare for credentialing.

Schooling and refugees : engaging with the complex trajectories of globalisation

This paper examines the complexities associated with educating a mobile and politically marginalised population, refugee students, in the state of Queensland, Australia. Historically, schools have been national institutions concerned with social reproduction and citizenship formation with a focus on spatially fixed populations. While education authorities in much of the developed world now acknowledge the need to prepare students for a more interconnected world of work and opportunity, they have largely failed to provide systemic support for one category of children on the move - refugees. We begin this paper with a discussion of forced migration and its links with ‘globalisation’. We then present our research findings about the educational challenges confronting individual refugee youth and schools in Queensland. This is followed with a summary of good practice in refugee education. The paper concludes with a discussion of how nation-states might play a more active role in facilitating transitions to citizenship for refugee youth.

Constructability improvement in sea water intake structure

Purpose The purpose of this paper is to explore the process, and analyse the implementation of constructability improvement and innovation result during the planning and design for sea water intake structure of fertilizer plant project. Design/methodology/approach The research methodology approach is case study method at project level. This constructability improvement process was investigated by using constructability implementation check lists, direct observation, documented lesson learned analysis and key personnel interviews. Findings The case study shows that the implementation of constructability during planning and design stage for this sea water intake structure has increased the project performance as well as improved the schedule by 5 months (14.21%) and reduced the project cost by 15.35%. Research limitations/implications This case study was limited to three (3) previous sea water intake projects as references and one (1) of new method sea water intake structure at fertilizer plant project. Practical implications A constructability improvement check list using theory and lesson learned for the specific construction project was documented. Originality/value The findings support the relevant study of constructability and provide specific lesson learned for three (3) previous project and one (1) of the new innovation method of the construction project and documented by the company.

Virion : new media and the development of the discursive museum

The historical rhetoric established with the very first public art museums declared that the purpose of such institutions was to provide a space where art could be accessible to all citizens. However contrary to this aim, studies show that art museums are one of the least accessed cultural institutions in the western world. The prevailing consensus for this can be attributed to the perception that museums are elitist, irrelevant and restricted to a small and privileged group. The focus of this research project is to address the issues that lead to these perceptions, and to identify possible curatorial strategies to encourage greater access to, and participation in the visual arts. This will be done through designing and curating an open submission exhibition that utilises new media technologies to increase access and dialogue between artists and audiences. This is part of a hybrid practice-based methodology that also includes scholarly research to critically investigate a number of historical and contemporary theories concerned with public museums and approaches to curatorial practice. This research will culminate in the development of Virion, an Internet based exhibition that aims to develop a curatorial model that facilitates open and democratic participation in arts practice from a diverse public audience.

Inhibition of p38 pathway leads to OA-like changes in a rat animal model

Objectives The p38 mitogen-activated protein kinase (MAPK) signal transduction pathway is involved in a variety of inflammatory responses, including cytokine generation, cell differentiation proliferation and apoptosis. Here, we examined the effects of systemic p38 MAPK inhibition on cartilage cells and osteoarthritis (OA) disease progression by both in vitro and in vivo approaches. Methods p38 kinase activity was evaluated in normal and OA cartilage cells by measuring the amount of phosphorylated protein. To examine the function of p38 signaling pathway in vitro, normal chondrocytes were isolated and differentiated in the presence or absence of p38 inhibitor; SB203580 and analysed for chondrogenic phenotype. Effect of systemic p38 MAPK inhibition in normal and OA (induced by menisectomy) rats were analysed by treating animals with vehicle alone (DMS0) or p38 inhibitor (SB203580). Damage to the femur and tibial plateau was evaluated by modified Mankin score, histology and immunohistochemistry. Results Our in vitro studies have revealed that a down-regulation of chondrogenic and increase of hypertrophic gene expression occurs in the normal chondrocytes, when p38 is neutralized by a pharmacological inhibitor. We further observed that the basal levels of p38 phosphorylation were decreased in OA chondrocytes compared with normal chondrocytes. These findings together indicate the importance of this pathway in the regulation of cartilage physiology and its relevance to OA pathogenesis. At in vivo level, systematic administration of a specific p38 MAPK inhibitor, SB203580, continuously for over a month led to a significant loss of proteoglycan; aggrecan and cartilage thickness. On the other hand, SB203580 treated normal rats showed a significant increase in TUNEL positive cells, cartilage hypertrophy markers such as Type 10 collagen, Runt-related transcription factor and Matrix metalloproteinase-13 and substantially induced OA like phenotypic changes in the normal rats. In addition, menisectomy induced OA rat models that were treated with p38 inhibitor showed aggravation of cartilage damage. Conclusions In summary, this study has provided evidence that the component of the p38 MAPK pathway is important to maintain the cartilage health and its inhibition can lead to severe cartilage degenerative changes. The observations in this study highlight the possibility of using activators of the p38 pathway as an alternative approach in the treatment of OA.