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.

Grounded theory : a methodological spiral from positivism to postmodernism

Aim. Our aim in this paper is to explain a methodological/methods package devised to incorporate situational and social world mapping with frame analysis, based on a grounded theory study of Australian rural nurses' experiences of mentoring. Background. Situational analysis, as conceived by Adele Clarke, shifts the research methodology of grounded theory from being located within a postpositivist paradigm to a postmodern paradigm. Clarke uses three types of maps during this process: situational, social world and positional, in combination with discourse analysis. Method. During our grounded theory study, the process of concurrent interview data generation and analysis incorporated situational and social world mapping techniques. An outcome of this was our increased awareness of how outside actors influenced participants in their constructions of mentoring. In our attempts to use Clarke's methodological package, however, it became apparent that our constructivist beliefs about human agency could not be reconciled with the postmodern project of discourse analysis. We then turned to the literature on symbolic interactionism and adopted frame analysis as a method to examine the literature on rural nursing and mentoring as secondary form of data. Findings. While we found situational and social world mapping very useful, we were less successful in using positional maps. In retrospect, we would argue that collective action framing provides an alternative to analysing such positions in the literature. This is particularly so for researchers who locate themselves within a constructivist paradigm, and who are therefore unwilling to reject the notion of human agency and the ability of individuals to shape their world in some way. Conclusion. Our example of using this package of situational and social worlds mapping with frame analysis is intended to assist other researchers to locate participants more transparently in the social worlds that they negotiate in their everyday practice. © 2007 Blackwell Publishing Ltd.

The ecological dynamics of decision-making in sailing

Competitive sailing is characterised by continuous interdependencies of decisions and actions. All actions imply a permanent monitoring of the environmental conditions, such as intensity and direction of the wind, sea characteristics, and the behaviour of the opponent sailors. These constraints on sailors’ behavior are in constant change implying continuous adjustments in sailors’ actions and decisions. Among the different parts of a regatta, tactics and strategy at the start are particularly relevant. Among coaches there is an adage that says that “the start is 50% of a regatta” (Houghton, 1984; Saltonstall, 1983/1986). Olympic sailing regattas are performed with boats of the same class, by one, two or three sailors, depending on the boat class. Normally before the start, sailors visit the racing venue and analyse wind and sea characteristics, in order to fine- tune their boats accordingly. Then, five minutes before the start, sailors initiate starting procedures in order to be in a favourable position at the starting line (at the “second zero”). This position is selected during the start period according to wind shifts tendencies and the actions of other boats (Figure 11.1). Only after the start signal can the boats cross the imaginary starting line between the race committee signal boat “A” and the pin end boat. The start takes place against the wind (upwind), and the boats start racing in the direction of mark 1. Based on the evaluation of the sea and wind characteristics (e.g. if the wind is stronger at a particular place on the course), sailors re- adjust their strategy for the regatta. This strategy may change during the regatta, according to wind changes and adversary actions. More to the point, strategic decisions constrain and are constrained by on- line decisions during the regatta.

Influence of culture conditions on the molecular signature of mesenchymal stem cells

Cell based therapies require cells capable of self renewal and differentiation, and a prerequisite is the ability to prepare an effective dose of ex vivo expanded cells for autologous transplants. The in vivo identification of a source of physiologically relevant cell types suitable for cell therapies is therefore an integral part of tissue engineering. Bone marrow is the most easily accessible source of mesenchymal stem cells (MSCs), and harbours two distinct populations of adult stem cells; namely hematopoietic stem cells (HSCs) and bone mesenchymal stem cells (BMSCs). Unlike HSCs, there are yet no rigorous criteria for characterizing BMSCs. Changing understanding about the pluripotency of BMSCs in recent studies has expanded their potential application; however, the underlying molecular pathways which impart the features distinctive to BMSCs remain elusive. Furthermore, the sparse in vivo distribution of these cells imposes a clear limitation to their in vitro study. Also, when BMSCs are cultured in vitro there is a loss of the in vivo microenvironment which results in a progressive decline in proliferation potential and multipotentiality. This is further exacerbated with increased passage number, characterized by the onset of senescence related changes. Accordingly, establishing protocols for generating large numbers of BMSCs without affecting their differentiation potential is necessary. The principal aims of this thesis were to identify potential molecular factors for characterizing BMSCs from osteoarthritic patients, and also to attempt to establish culture protocols favourable for generating large number of BMSCs, while at the same time retaining their proliferation and differentiation potential. Previously published studies concerning clonal cells have demonstrated that BMSCs are heterogeneous populations of cells at various stages of growth. Some cells are higher in the hierarchy and represent the progenitors, while other cells occupy a lower position in the hierarchy and are therefore more committed to a particular lineage. This feature of BMSCs was made evident by the work of Mareddy et al., which involved generating clonal populations of BMSCs from bone marrow of osteoarthritic patients, by a single cell clonal culture method. Proliferation potential and differentiation capabilities were used to group cells into fast growing and slow growing clones. The study presented here is a continuation of the work of Mareddy et al. and employed immunological and array based techniques to identify the primary molecular factors involved in regulating phenotypic characteristics exhibited by contrasting clonal populations. The subtractive immunization (SI) was used to generate novel antibodies against favourably expressed proteins in the fast growing clonal cell population. The difference between the clonal populations at the transcriptional level was determined using a Stem Cell RT2 Profiler TM PCR Array which focuses on stem cell pathway gene expression. Monoclonal antibodies (mAb) generated by SI were able to effectively highlight differentially expressed antigenic determinants, as was evident by Western blot analysis and confocal microscopy. Co-immunoprecipitation, followed by mass spectroscopy analysis, identified a favourably expressed protein as the cytoskeletal protein vimentin. The stem cell gene array highlighted genes that were highly upregulated in the fast growing clonal cell population. Based on their functions these genes were grouped into growth factors, cell fate determination and maintenance of embryonic and neural stem cell renewal. Furthermore, on a closer analysis it was established that the cytoskeletal protein vimentin and nine out of ten genes identified by gene array were associated with chondrogenesis or cartilage repair, consistent with the potential role played by BMSCs in defect repair and maintaining tissue homeostasis, by modulating the gene expression pattern to compensate for degenerated cartilage in osteoarthritic tissues. The gene array also presented transcripts for embryonic lineage markers such as FOXA2 and Sox2, both of which were significantly over expressed in fast growing clonal populations. A recent groundbreaking study by Yamanaka et al imparted embryonic stem cell (ESCs) -like characteristic to somatic cells in a process termed nuclear reprogramming, by the ectopic expression of the genes Sox2, cMyc and Oct4. The expression of embryonic lineage markers in adult stem cells may be a mechanism by which the favourable behaviour of fast growing clonal cells is determined and suggests a possible active phenomenon of spontaneous reprogramming in fast growing clonal cells. The expression pattern of these critical molecular markers could be indicative of the competence of BMSCs. For this reason, the expression pattern of Sox2, Oct4 and cMyc, at various passages in heterogeneous BMSCs population and tissue derived cells (osteoblasts and chondrocytes), was investigated by a real-time PCR and immunoflourescence staining. A strong nuclear staining was observed for Sox2, Oct4 and cMyc, which gradually weakened accompanied with cytoplasmic translocation after several passage. The mRNA and protein expression of Sox2, Oct4 and cMyc peaked at the third passage for osteoblasts, chondrocytes and third passage for BMSCs, and declined with each subsequent passage, indicating towards a possible mechanism of spontaneous reprogramming. This study proposes that the progressive decline in proliferation potential and multipotentiality associated with increased passaging of BMSCs in vitro might be a consequence of loss of these propluripotency factors. We therefore hypothesise that the expression of these master genes is not an intrinsic cell function, but rather an outcome of interaction of the cells with their microenvironment; this was evident by the fact that when removed from their in vivo microenvironment, BMSCs undergo a rapid loss of stemness after only a few passages. One of the most interesting aspects of this study was the integration of factors in the culture conditions, which to some extent, mimicked the in vivo microenvironmental niche of the BMSCs. A number of studies have successfully established that the cellular niche is not an inert tissue component but is of prime importance. The total sum of stimuli from the microenvironment underpins the complex interplay of regulatory mechanisms which control multiple functions in stem cells most importantly stem cell renewal. Therefore, well characterised factors which affect BMSCs characteristics, such as fibronectin (FN) coating, and morphogens such as FGF2 and BMP4, were incorporated into the cell culture conditions. The experimental set up was designed to provide insight into the expression pattern of the stem cell related transcription factors Sox2, cMyc and Oct4, in BMSCs with respect to passaging and changes in culture conditions. Induction of these pluripotency markers in somatic cells by retroviral transfection has been shown to confer pluripotency and an ESCs like state. Our study demonstrated that all treatments could transiently induce the expression of Sox2, cMyc and Oct4, and favourably affect the proliferation potential of BMSCs. The combined effect of these treatments was able to induce and retain the endogenous nuclear expression of stem cell transcription factors in BMSCs over an extended number of in vitro passages. Our results therefore suggest that the transient induction and manipulation of endogenous expression of transcription factors critical for stemness can be achieved by modulating the culture conditions; the benefit of which is to circumvent the need for genetic manipulations. In summary, this study has explored the role of BMSCs in the diseased state of osteoarthritis, by employing transcriptional profiling along with SI. In particular this study pioneered the use of primary cells for generating novel antibodies by SI. We established that somatic cells and BMSCs have a basal level of expression of pluripotency markers. Furthermore, our study indicates that intrinsic signalling mechanisms of BMSCs are intimately linked with extrinsic cues from the microenvironment and that these signals appear to be critical for retaining the expression of genes to maintain cell stemness in long term in vitro culture. This project provides a basis for developing an “artificial niche” required for reversion of commitment and maintenance of BMSC in their uncommitted homeostatic state.

Ceremony and cybrary : Digital libraries and the dialectic of place and space

This article examines social, cultural and technological change in the systems and economies of educational information management. Since the Sumerians first collected, organized and supervised administrative and religious records some six millennia ago, libraries have been key physical depositories and cultural signifiers in the production and mediation of social capital and power through education. To date, the textual, archival and discursive practices perpetuating libraries have remained exempt from inquiry. My aim here is to remedy this hiatus by making the library itself the terrain and object of critical analysis and investigation. The paper argues that in the three dominant communications eras—namely, oral, print and digital cultures—society’s centres of knowledge and learning have resided in the ceremony, the library and the cybrary respectively. In a broad-brush historical grid, each of these key educational institutions—the ceremony in oral culture, the library in print culture and the cybrary in digital culture—are mapped against social, cultural and technological orders pertaining to their era. Following a description of these shifts in society’s collective cultural memory, the paper then examines the question of what the development of global information systems and economies mean for schools and libraries of today, and for teachers and learners as knowledge consumers and producers?

Diversity in action : minority group media and social change

My thesis consists of a creative work plus an exegesis. This exegesis uses case study research to investigate three Brisbane-based media organisations and the role they play in encouraging social inclusion and other positive social change for specific disadvantaged and stigmatised minority groups. Bailey, Cammaerts and Carpentier’s theoretical approach to alternative media forms the basis of this research. Bailey et al. (2008, p. 156) view alternative media organisations as having four important roles, two media-centred and two society-centred, which must all be considered to best understand them: • serving their communities • acting as an alternative to mainstream media discourses • promoting and advocating democratisation in the media and through the media in society • functioning as a crossroads in civil society. The first case study, about community radio station 4RPH (Radio for the Print Handicapped), centres on promoting social inclusion for people with a print disability through access to printed materials (primarily mainstream print media) in an audio format. The station also provides important opportunities for members of this group to produce media and, to a lesser extent, provides disability-specific information and discussions. The second case study, about gay print and online magazine Queensland Pride, focuses on promoting social inclusion and combating the discrimination and repression of people who identify as lesbian, gay, bisexual or transgender. Central issues include the representation (including sexualised representation) of a subculture and niche target market, and the impact of commercialisation on this free publication. The third case study, about community radio station 98.9FM, explores the promotion of social inclusion for peoples whose identity, cultures, issues, politics and contributions are often absent or misrepresented in the mainstream media. This radio station provides “a first level of service” (Meadows & van Vuuren, 1998, p. 104) to these people, but also informs and entertains those in the majority society. The findings of this research suggest that there are two key mechanisms that help these media organisations to effect social change: first, strengthening the minority community and serving its needs, and second, fostering connections with the broader society.

Reversed bias Pt/nanostructured ZnO Schottky diode with enhanced electric field for hydrogen sensing

In this paper, the effect of electric field enhancement on Pt/nanostructured ZnO Schottky diode based hydrogen sensors under reverse bias condition has been investigated. Current-voltage characteristics of these diodes have been studied at temperatures from 25 to 620 °C and their free carrier density concentration was estimated by exposing the sensors to hydrogen gas. The experimental results show a significantly lower breakdown voltage in reversed bias current-voltage characteristics than the conventional Schottky diodes and also greater lateral voltage shift in reverse bias operation than the forward bias. This can be ascribed to the increased localized electric fields emanating from the sharp edges and corners of the nanostructured morphologies. At 620 °C, voltage shifts of 114 and 325 mV for 0.06% and 1% hydrogen have been recorded from dynamic response under the reverse bias condition. © 2010 Elsevier B.V. All rights reserved.

A plasmonic 'ac Wheatstone bridge' circuit for high-sensitivity phase measurement and single-molecule detection

In this paper, a plasmonic “ac Wheatstone bridge” circuit is proposed and theoretically modeled for the first time. The bridge circuit consists of three metallic nanoparticles, shaped as rectangular prisms, with two nanoparticles acting as parallel arms of a resonant circuit and the third bridging nanoparticle acting as an optical antenna providing an output signal. Polarized light excites localized surface plasmon resonances in the two arms of the circuit, which generate an optical signal dependent on the phase-sensitive excitations of surface plasmons in the antenna. The circuit is analyzed using a plasmonic coupling theory and numerical simulations. The analyses show that the plasmonic circuit is sensitive to phase shifts between the arms of the bridge and has the potential to detect the presence of single molecules.

Position, rotation, and scale invariant recognition of images using higher-order spectra

A new approach to recognition of images using invariant features based on higher-order spectra is presented. Higher-order spectra are translation invariant because translation produces linear phase shifts which cancel. Scale and amplification invariance are satisfied by the phase of the integral of a higher-order spectrum along a radial line in higher-order frequency space because the contour of integration maps onto itself and both the real and imaginary parts are affected equally by the transformation. Rotation invariance is introduced by deriving invariants from the Radon transform of the image and using the cyclic-shift invariance property of the discrete Fourier transform magnitude. Results on synthetic and actual images show isolated, compact clusters in feature space and high classification accuracies

From margins to mainstream : how screen and creative industries developed in the Northern Rivers region of NSW : 2000–2010

This thesis explores the proposition that growth and development in the screen and creative industries is not confined to the major capital cities. Lifestyle considerations, combined with advances in digital technology, convergence and greater access to broadband are altering requirements for geographic location, and creative workers are being drawn away from the big metropolises to certain regional areas. Regional screen industry enclaves are emerging outside of London, in the Highlands and Islands of Scotland, in Nova Scotia in Canada and in New Zealand. In the Australian context, the proposition is tested in an area regarded as a ‘special case’ in creative industry expansion: the Northern Rivers region of NSW. A key feature of the ‘specialness’ of this region is the large number of experienced, credited producers who live and operate their businesses within the region. The development of screen and creative industries in the Northern Rivers over the decade 2000 – 2010 has implications for regional regeneration and offers new insights into the rapidly changing screen industry landscape. This development also has implications for creative industry discourse, especially the dominance of the urban in creative industries thought. The research is pioneering in a number of ways. Building on the work conducted for my Masters thesis in 2000, a second study was conducted during the research phase, adapting creative industries theory and mapping methods, which have been largely city and nation-centric, and applying them to a regional context. The study adopted an action research approach as an industry development strategy for screen industries, while at the same time developing fine-grained ground up methods for collecting primary quantitative data on the size and scope of the creative industries. In accordance with the action research framework, the researcher also acted in the dual roles of industry activist and screen industry producer in the region. The central focus of the research has been both to document and contribute to the growth and development of screen and creative industries over the past decade in the Northern Rivers region. These interventions, along with policy developments at both a local and national level, and broader global shifts, have had the effect of repositioning the sector from a marginal one to a priority area considered integral to the future economic and cultural life of the region. The research includes a detailed mapping study undertaken in 2005 with comparisons to an earlier 2000 study and to ABS data for 2001 and 2006 to reveal growth trends. It also includes two case studies of projects that developed from idea to production and completion in the region during the decade in question. The studies reveal the drivers, impediments and policy implications for sustaining the development of screen industries in a regional area. A major finding of the research was the large and increasing number of experienced producers who operate within the region and the leadership role they play in driving the development of the emerging local industry. The two case studies demonstrate the impact of policy decisions on local screen industry producers and their enterprises. A brief overview of research in other regional areas is presented, including two international examples, and what they reveal about regional regeneration. Implications are drawn for creative industries discourse and regional development policy challenges for the future.

Resisting contradictions : non-Indigenous pre-service teacher responses to critical Indigenous studies

The study examines non-Indigenous pre-service teacher responses to the authorisation of Indigenous knowledge perspectives in compulsory Indigenous studies with a primary focus on exploring the nature and effects of resistance. It draws on the philosophies of the Japanangka teaching and research paradigm (West, 2000), relationship theory (Graham, 1999), Indigenist methodologies and decolonisation approaches to examine this resistance. A Critical Indigenist Study was employed to investigate how non-Indigenous pre-service teachers managed their learning, and how they articulated shifts in resistance as they progressed through their studies. This study explains resistance to compulsory Indigenous and how it can be targeted by Indigenist Standpoint Pedagogy. The beginning transformations in pre-service teacher positioning in relation to Australian history, contemporary educational practice, and professional identity was also explored.

A hydrogen gas sensor based on Pt/nanostructured WO3/SiC Schottky diode

Pt/nanostructured WO3/SiC Schottky diodes were fabricated and applied for hydrogen gas sensing applications. The nanostructured WO3 films were synthesized from tungsten coated SiC substrates via an acid-etching method using a 1.5 M HNO3 solution for 1 hr, 2 hrs and 3 hrs duration. Scanning electron microscopy of the developed films revealed platelet crystals with thicknesses in the order of 20-60 nm and lengths between 100-700 nm. X-ray diffraction analysis revealed that the rate of oxidation of tungsten increases as the duration of acid-etching increases. The devices were tested towards hydrogen gas balanced in air at different temperatures from 25°C to 200°C. At 200°C, voltage shifts of 0.45 V, 0.93 V and 2.37 V were recorded for devices acid-etched for 1 hr, 2 hrs and 3 hrs duration, respectively upon exposure to 1% hydrogen, under a constant forward bias current of 500 µA.

Grocery product pricing and Australian supermarket consumers : gender differences in perceived importance levels

Grocery shopping is an essential and routine activity. Although long regarded the responsibility of the female spouse, modern social and demographic shifts are causing men to become more engaged in this task. This is the first study to analyse gender differences with respect to the criterion of grocery product price within an Australian supermarket retail environment. A stratified sample of 140 male and 140 female grocery shoppers was surveyed. Results showed that men considered price attributes of products as being significantly lower in importance than did women. Additionally, men displayed lower levels of price nvolvement, reported referencing shelf price to a lesser extent, and gave lesser consideration to promotional tactics focusing on low price. Although men on average buy fewer items than do women, they spend more money for each item they purchase. This higher expenditure per item appears to be driven, at least in part, by a lack of price referencing. This research has implications for gender studies and consumer behaviour disciplines in relation to grocery shopping.

The emergent male shopper : an identification of male supermarket shopper types

Grocery shopping is a routine activity widely considered the responsibility of the female spouse, yet modern social and demographic shifts are causing men to engage in this task. This study develops a retail shopping typology of male grocery shoppers, employing a cluster analysis technique. Five distinct cohorts emerge from the data of eight constructs, measured by seventy one items. One new shopper type emerges from this research. This shopper presented as a younger man, at the commencement of their family lifecycle, attracted by a strong value offer, focusing on price and promotional discounts. Our research offers a contribution to the marketing, consumer behaviour and supermarket retailing disciplines in three ways. By examining and identifying male shopping behaviour in the context of grocery shopping, the development of a retail shopping typology of male grocery shoppers and the extension and employment of a cluster analysis in identifying distinct groups. This research has implications for gender, segmentation studies and consumer behaviour disciplines in regard to grocery shopping. The identification of specific groups of male grocery shoppers will enable grocery retailers to effectively implement important, targeted marketing strategies.

International law and the regulation of violence : shifting perceptions of the legitimacy of the jus ad bellum?

International law’s capacity to influence state behaviour by regulating recourse to violence has been a longstanding source of debate among international lawyers and political scientists. On the one hand, sceptics assert that frequent violations of the prohibition on the use of force have rendered article 2(4) of the UN Charter redundant. They contend that national self-interest, rather than international law, is the key determinant of state behaviour regarding the use of force. On the other hand, defenders of article 2(4) argue first, that most states comply with the Charter framework, and second, that state rhetoric continues to acknowledge the existence of the jus ad bellum. In particular, the fact that violators go to considerable lengths to offer legal or factual justifications for their conduct – typically by relying on the right of self-defence – is advanced as evidence that the prohibition on the use of force retains legitimacy in the eyes of states. This paper identifies two potentially significant features of state practice since 2006 which may signal a shift in states’ perceptions of the normative authority of article 2(4). The first aspect is the recent failure by several states to offer explicit legal justifications for their use or force, or to report action taken in self-defence to the Security Council in accordance with Article 51. Four incidents linked to the global “war on terror” are examined here: Israeli airstrikes in Syria in 2007 and in Sudan in 2009, Turkey’s 2006-2008 incursions into northern Iraq, and Ethiopia’s 2006 intervention in Somalia. The second, more troubling feature is the international community’s apparent lack of concern over the legality of these incidents. Each use of force is difficult to reconcile with the strict requirements of the jus ad bellum; yet none attracted genuine legal scrutiny or debate among other states. While it is too early to conclude that these relatively minor incidents presage long term shifts in state practice, viewed together the two developments identified here suggest a possible downgrading of the role of international law in discussions over the use of force, at least in conflicts linked to the “war on terror”. This, in turn, may represent a declining perception of the normative authority of the jus ad bellum, and a concomitant admission of the limits of international law in regulating violence.