An analysis of the variability in Queensland WiM data

The Queensland Department of Main Roads uses Weigh-in-Motion (WiM) devices to covertly monitor (at highway speed) axle mass, axle configurations and speed of heavy vehicles on the road network. Such data is critical for the planning and design of the road network. Some of the data appears excessively variable. The current work considers the nature, magnitude and possible causes of WiM data variability. Over fifty possible causes of variation in WiM data have been identified in the literature. Data exploration has highlighted five basic types of variability specifically: ----- • cycling, both diurnal and annual;----- • consistent but unreasonable data;----- • data jumps;----- • variations between data from opposite sides of the one road; and ----- • non-systematic variations.----- This work is part of wider research into procedures to eliminate or mitigate the influence of WiM data variability.

Policing lesbian, gay, bisexual and transgender young people : a gap in the research literature

This article argues for exploring lesbian, gay, bisexual, and transgender (LGBT) young people’s experiences with police. While research examines how factors such as indigeneity influence young peoples’ experiences with police, how sexuality and/or gender identity mediates these relationships remains largely unexplored. Key bodies of research suggest a need to explore this area further, including: literature documenting links between homophobic violence against LGBT young people and outcomes such as homelessness that fall within the gambit of policing work; research showing reluctance of LGBT communities to report crime to police; international research documenting homophobic police attitudes and Australian research demonstrating arguably homophobic court outcomes; and research outlining increasing police support of LGBT communities. Drawing on these bodies of literature, this article argues that LGBT young people experience policing warrants further research.

Social network market : storytelling on a Web 2.0 original literature site

This article looks at a Chinese Web 2.0 original literature site, Qidian, in order to show the coevolution of market and non-market initiatives. The analytic framework of social network markets (Potts et al., 2008) is employed to analyse the motivations of publishing original literature works online and to understand the support mechanisms of the site, which encourage readers’ willingness to pay for user-generated content. The co-existence of socio-cultural and commercial economies and their impact on the successful business model of the site are illustrated in this case. This article extends the concept of social network markets by proposing the existence of a ripple effect of social network markets through convergence between PC and mobile internet, traditional and internet publishing, and between publishing and other cultural industries. It also examines the side effects of social network markets, and the role of market and non-market strategies in addressing the issues.

The influence of underreported crashes on hotspot identification

Hot spot identification (HSID) plays a significant role in improving the safety of transportation networks. Numerous HSID methods have been proposed, developed, and evaluated in the literature. The vast majority of HSID methods reported and evaluated in the literature assume that crash data are complete, reliable, and accurate. Crash under-reporting, however, has long been recognized as a threat to the accuracy and completeness of historical traffic crash records. As a natural continuation of prior studies, the paper evaluates the influence that under-reported crashes exert on HSID methods. To conduct the evaluation, five groups of data gathered from Arizona Department of Transportation (ADOT) over the course of three years are adjusted to account for fifteen different assumed levels of under-reporting. Three identification methods are evaluated: simple ranking (SR), empirical Bayes (EB) and full Bayes (FB). Various threshold levels for establishing hotspots are explored. Finally, two evaluation criteria are compared across HSID methods. The results illustrate that the identification bias—the ability to correctly identify at risk sites--under-reporting is influenced by the degree of under-reporting. Comparatively speaking, crash under-reporting has the largest influence on the FB method and the least influence on the SR method. Additionally, the impact is positively related to the percentage of the under-reported PDO crashes and inversely related to the percentage of the under-reported injury crashes. This finding is significant because it reveals that despite PDO crashes being least severe and costly, they have the most significant influence on the accuracy of HSID.

Does institutional location protect from political influence? the case of a minimum labour standards enforcement agency in Australia

Among the many factors that influence enforcement agencies, this article examines the role of the institutional location (and independence) of agencies, and an incumbent government's ideology. It is argued that institutional location affects the level of political influence on the agency's operations, while government ideology affects its willingness to resource enforcement agencies and approve regulatory activities. Evidence from the agency regulating minimum labour standards in the Australian federal industrial relations jurisdiction (currently the Fair Work Ombudsman) highlights two divergences from the regulatory enforcement literature generally. First, notions of independence from political interference offered by institutional location are more illusory than real and, second, political need motivates political action to a greater extent than political ideology.

Unpacking Japan's 21st century "National Conversation" : images of the future beyond the iron cage of the "Catch Up" model

How does the image of the future operate upon history, and upon national and individual identities? To what extent are possible futures colonized by the image? What are the un-said futurecratic discourses that underlie the image of the future? Such questions inspired the examination of Japan’s futures images in this thesis. The theoretical point of departure for this examination is Polak’s (1973) seminal research into the theory of the ‘image of the future’ and seven contemporary Japanese texts which offer various alternative images for Japan’s futures, selected as representative of a ‘national conversation’ about the futures of that nation. These seven images of the future are: 1. Report of the Prime Minister’s Commission on Japan’s Goals in the 21st Century—The Frontier Within: Individual Empowerment and Better Governance in the New Millennium, compiled by a committee headed by Japan’s preeminent Jungian psychologist Kawai Hayao (1928-2007); 2. Slow Is Beautiful—a publication by Tsuji Shinichi, in which he re-images Japan as a culture represented by the metaphor of the sloth, concerned with slow and quality-oriented livingry as a preferred image of the future to Japan’s current post-bubble cult of speed and economic efficiency; 3. MuRatopia is an image of the future in the form of a microcosmic prototype community and on-going project based on the historically significant island of Awaji, and established by Japanese economist and futures thinker Yamaguchi Kaoru; 4. F.U.C.K, I Love Japan, by author Tanja Yujiro provides this seven text image of the future line-up with a youth oriented sub-culture perspective on that nation’s futures; 5. IMAGINATION / CREATION—a compilation of round table discussions about Japan’s futures seen from the point of view of Japan’s creative vanguard; 6. Visionary People in a Visionless Country: 21 Earth Connecting Human Stories is a collection of twenty one essays compiled by Denmark born Tokyo resident Peter David Pedersen; and, 7. EXODUS to the Land of Hope, authored by Murakami Ryu, one of Japan’s most prolific and influential writers, this novel suggests a future scenario portraying a massive exodus of Japan’s youth, who, literate with state-of-the-art information and communication technologies (ICTs) move en masse to Japan’s northern island of Hokkaido to launch a cyber-revolution from the peripheries. The thesis employs a Futures Triangle Analysis (FTA) as the macro organizing framework and as such examines both pushes of the present and weights from the past before moving to focus on the pulls to the future represented by the seven texts mentioned above. Inayatullah’s (1999) Causal Layered Analysis (CLA) is the analytical framework used in examining the texts. Poststructuralist concepts derived primarily from the work of Michel Foucault are a particular (but not exclusive) reference point for the analytical approach it encompasses. The research questions which reflect the triangulated analytic matrix are: 1. What are the pushes—in terms of current trends—that are affecting Japan’s futures? 2. What are the historical and cultural weights that influence Japan’s futures? 3. What are the emerging transformative Japanese images of the future discourses, as embodied in actual texts, and what potential do they offer for transformative change in Japan? Research questions one and two are discussed in Chapter five and research question three is discussed in Chapter six. The first two research questions should be considered preliminary. The weights outlined in Chapter five indicate that the forces working against change in Japan are formidable, structurally deep-rooted, wide-spread, and under-recognized as change-adverse. Findings and analyses of the push dimension reveal strong forces towards a potentially very different type of Japan. However it is the seven contemporary Japanese images of the future, from which there is hope for transformative potential, which form the analytical heart of the thesis. In analyzing these texts the thesis establishes the richness of Japan’s images of the future and, as such, demonstrates the robustness of Japan’s stance vis-à-vis the problem of a perceived map-less and model-less future for Japan. Frontier is a useful image of the future, whose hybrid textuality, consisting of government, business, academia, and creative minority perspectives, demonstrates the earnestness of Japan’s leaders in favour of the creation of innovative futures for that nation. Slow is powerful in its aim to reconceptualize Japan’s philosophies of temporality, and build a new kind of nation founded on the principles of a human-oriented and expanded vision of economy based around the core metaphor of slowness culture. However its viability in Japan, with its post-Meiji historical pushes to an increasingly speed-obsessed social construction of reality, could render it impotent. MuRatopia is compelling in its creative hybridity indicative of an advanced IT society, set in a modern day utopian space based upon principles of a high communicative social paradigm, and sustainability. IMAGINATION / CREATION is less the plan than the platform for a new discussion on Japan’s transformation from an econo-centric social framework to a new Creative Age. It accords with emerging discourses from the Creative Industries, which would re-conceive of Japan as a leading maker of meaning, rather than as the so-called guzu, a term referred to in the book meaning ‘laggard’. In total, Love Japan is still the most idiosyncratic of all the images of the future discussed. Its communication style, which appeals to Japan’s youth cohort, establishes it as a potentially formidable change agent in a competitive market of futures images. Visionary People is a compelling image for its revolutionary and subversive stance against Japan’s vision-less political leadership, showing that it is the people, not the futures-making elite or aristocracy who must take the lead and create a new vanguard for the nation. Finally, Murakami’s Exodus cannot be ruled out as a compelling image of the future. Sharing the appeal of Tanja’s Love Japan to an increasingly disenfranchised youth, Exodus portrays a near-term future that is achievable in the here and now, by Japan’s teenagers, using information and communications technologies (ICTs) to subvert leadership, and create utopianist communities based on alternative social principles. The principal contribution from this investigation in terms of theory belongs to that of developing the Japanese image of the future. In this respect, the literature reviews represent a significant compilation, specifically about Japanese futures thinking, the Japanese image of the future, and the Japanese utopia. Though not exhaustive, this compilation will hopefully serve as a useful starting point for future research, not only for the Japanese image of the future, but also for all image of the future research. Many of the sources are in Japanese and their English summations are an added reason to respect this achievement. Secondly, the seven images of the future analysed in Chapter six represent the first time that Japanese image of the future texts have been systematically organized and analysed. Their translation from Japanese to English can be claimed as a significant secondary contribution. What is more, they have been analysed according to current futures methodologies that reveal a layeredness, depth, and overall richness existing in Japanese futures images. Revealing this image-richness has been one of the most significant findings of this investigation, suggesting that there is fertile research to be found from this still under-explored field, whose implications go beyond domestic Japanese concerns, and may offer fertile material for futures thinkers and researchers, Japanologists, social planners, and policy makers.

A literature review on research methodologies of gross pollutant traps

This paper presents a comprehensive review of scientific and grey literature on gross pollutant traps (GPTs). GPTs are designed with internal screens to capture gross pollutants—organic matter and anthropogenic litter. Their application involves professional societies, research organisations, local city councils, government agencies and the stormwater industry—often in partnership. In view of this, the 113 references include unpublished manuscripts from these bodies along with scientific peer-reviewed conference papers and journal articles. The literature reviewed was organised into a matrix of six main devices and nine research areas (testing methodologies) which include: design appraisal study, field monitoring/testing, experimental flow fields, gross pollutant capture/retention characteristics, residence time calculations, hydraulic head loss, screen blockages, flow visualisations and computational fluid dynamics (CFD). When the fifty-four item matrix was analysed, twenty-eight research gaps were found in the tabulated literature. It was also found that the number of research gaps increased if only the scientific literature was considered. It is hoped, that in addition to informing the research community at QUT, this literature review will also be of use to other researchers in this field.

Direct instruction in mathematics : issues for schools with high Indigenous enrolments : a literature review

Direct instruction, an approach that is becoming familiar to Queensland schools that have high Aboriginal and Torres Strait Islander populations, has been gaining substantial political and popular support in the United States of America [USA], England and Australia. Recent examples include the No Child Left Behind policy in the USA, the British National Numeracy Strategy and in Australia, Effective Third Wave Intervention Strategies. Direct instruction, stems directly from the model created in the 1960s under a Project Follow Through grant. It has been defined as a comprehensive system of education involving all aspects of instruction. Now in its third decade of influencing curriculum, instruction and research, direct instruction is also into its third decade of controversy because of its focus on explicit and highly directed instruction for learning. Characteristics of direct instruction are critiqued and discussed to identify implications for teaching and learning for Indigenous students.

Integrating information literacy into higher education curricula : an IL curricular integration model

This study investigates a way to systematically integrate information literacy (IL) into an undergraduate academic programme and develops a model for integrating information literacy across higher education curricula. Curricular integration of information literacy in this study means weaving information literacy into an academic curriculum. In the associated literature, it is also referred to as the information literacy embedding approach or the intra-curricular approach. The key findings identified from this study are presented in 4 categories: the characteristics of IL integration; the key stakeholders in IL integration; IL curricular design strategies; and the process of IL curricular integration. Three key characteristics of the curricular integration of IL are identified: collaboration and negotiation, contextualisation and ongoing interaction with information. The key stakeholders in the curricular integration of IL are recognised as the librarians, the course coordinators and lecturers, the heads of faculties or departments, and the students. Some strategies for IL curricular design include: the use of IL policies and standards in IL curricular design; the combination of face to face and online teaching as an emerging trend; the use of IL assessment tools which play an important role in IL integration. IL can be integrated into the intended curriculum (what an institution expects its students to learn), the offered curriculum (what the teachers teach) and the received curriculum (what students actually learn). IL integration is a process of negotiation, collaboration and the implementation of the intended curriculum. IL can be integrated at different levels of curricula such as: institutional, faculty, departmental, course and class curriculum levels. Based on these key findings, an IL curricular integration model is developed. The model integrates curriculum, pedagogy and learning theories, IL theories, IL guidelines and the collaboration of multiple partners. The model provides a practical approach to integrating IL into multiple courses across an academic degree. The development of the model was based on the IL integration experiences of various disciplines in three universities and the implementation experience of an engineering programme at another university; thus it may be of interest to other disciplines. The model has the potential to enhance IL teaching and learning, curricular development and to implement graduate attributes in higher education. Sociocultural theories are applied to the research process and IL curricular design of this study. Sociocultural theories describe learning as being embedded within social events and occurring as learners interact with other people, objects, and events in a collaborative environment. Sociocultural theories are applied to explore how academic staff and librarians experience the curricular integration of IL; they also support collaboration in the curricular integration of IL and the development of an IL integration model. This study consists of two phases. Phase I (2007) was the interview phase where both academic staff and librarians at three IL active universities were interviewed. During this phase, attention was paid specifically to the practical process of curricular integration of IL and IL activity design. Phase II, the development phase (2007-2008), was conducted at a fourth university. This phase explores the systematic integration of IL into an engineering degree from Year 1 to Year 4. Learning theories such as sociocultural theories, Bloom’s Taxonomy and IL theories are used in IL curricular development. Based on the findings from both phases, an IL integration model was developed. The findings and the model contribute to IL education, research and curricular development in higher education. The sociocultural approach adopted in this study also extends the application of sociocultural theories to the IL integration process and curricular design in higher education.

Influence of parameters on the heterogeneous photocatalytic degradation of pesticides and phenolic contaminants in wastewater : a short review

In recent years, the application of heterogeneous photocatalytic water purification process has gained wide attention due to its effectiveness in degrading and mineralizing the recalcitrant organic compounds as well as the possibility of utilizing the solar UV and visible light spectrum. This paper aims to review and summarize the recently published works on the titanium dioxide (TiO2) photocatalytic oxidation of pesticides and phenolic compounds, predominant in storm and waste water effluents. The effect of various operating parameters on the photocatalytic degradation of pesticides and phenols are discussed. Results reported here suggested that the photocatalytic degradation of organic compounds depends on the type of photocatalyst and composition, light intensity, initial substrate concentration, amount of catalyst, pH of the reaction medium, ionic components in water, solvent types, oxidizing agents/electron acceptors, catalyst application mode, and calcinations temperature in water environment. A substantial amount of research has focused on the enhancement of TiO2 photocatalysis by modification with metal, non-metal and ion doping. Recent developments in TiO2 photocatalysis for the degradation of various pesticides and phenols are also highlighted in this review. It is evident from the literature survey that photocatalysis has shown good potential for the removal of various organic pollutants. However, still there is a need to find out the practical utility of this technique on commercial scale.

Fatalism and road safety in developing countries, with a focus on Pakistan

Road crashes are a significant problem in developing countries such as Pakistan. Attitudes are among the human factors which influence risky road use and receptiveness to interventions. Fatalism is a set of attitudes known to be important in Pakistan and other developing countries; however it is rarely addressed in the road safety literature. Two broad types of fatalism are “theological fatalism” and “empirical fatalism”, both of which are found in developed countries as well as in developing countries. Where research has been conducted into the issue, fatalism is considered to interfere with messages aimed at improving road safety. Pakistan has a serious road crash problem, and there is sufficient information to suggest that fatalism is an important contributing factor to the problem, but a better understanding of how fatalism operates in Pakistan is needed if effective prevention strategies are to be developed. A proposed study using an anthropological approach is described which will be exploratory in nature and which is aimed at investigating fatalism and related concepts among Pakistani road users and those who develop and implement road safety policy.

Mapping the ethical journey of experienced nurses now practising in rural and remote hospitals in central and south-west Queensland and in domiciliary services in Brisbane : a grounded theory approach

The aim of this thesis has been to map the ethical journey of experienced nurses now practising in rural and remote hospitals in central and south-west Queensland and in domiciliary services in Brisbane. One group of the experienced nurses in the study were Directors of Nursing in rural and remote hospitals. These nurses were “hands on”, “multi-skilled “ nurses who also had the task of managing the hospital. Also there were two Directors of Nursing from domiciliary services in Brisbane. A grounded theory method was used. The nurses were interviewed and the data retrieved from the interviews was coded, categorised and from these categories a conceptual framework was generated. The literature which dealt with the subject of ethical decision making and nurses also became part of the data. The study revealed that all these nurses experienced moral distress as they made ethical decisions. The decision making categories revealed in the data were: the area of financial management; issues as end of life approaches; allowing to die with dignity; emergency decisions; experience of unexpected death; the dilemma of providing care in very difficult circumstances. These categories were divided into two chapters: the category related to administrative and financial constraints and categories dealing with ethical issues in clinical settings. A further chapter discussed the overarching category of coping with moral distress. These experienced nurses suffered moral distress as they made ethical decisions, confirming many instances of moral distress in ethical decision making documented in the literature to date. Significantly, the nurses in their interviews never mentioned the ethical principles used in bioethics as an influence in their decision making. Only one referred to lectures on ethics as being an influence in her thinking. As they described their ethical problems and how they worked through them, they drew on their own previous experience rather than any knowledge of ethics gained from nursing education. They were concerned for their patients, they spoke from a caring responsibility towards their patients, but they were also concerned for justice for their patients. This study demonstrates that these nurses operated from the ethic of care, tempered with the ethic of responsibility as well as a concern for justice for their patients. Reflection on professional experience, rather than formal ethics education and training, was the primary influence on their ethical decision making.

Inter-sector leader transitions and organisation transformation : a research agenda

Faced with the need for strategic change, structural and cultural realignment, innovation and value-adding, many public sector organisations are tapping into a wider senior leadership talent pool and attracting successful leaders from other sectors (Flynn and Thompson, 2009). Leadership renewal has resulted, in some cases, in the external recruitment of whole senior leadership teams (Hockridge, 2008), raising issues about the influence of context on leader success (Pawar and Eastman, 1997) and potential leader transition failure, a costly outcome for leaders and organisations (Howard, 2001). There is little research on inter-sector leader transitions, which is surprising given the significant costs associated with leader acquisition and failure(Conger, 2010; Day and Halpin, 2004). For example, it is not clear what organizations do (or do not do) to ensure the outcomes of their significant investment in inter sector transitions are realised. In addition, it is not clear how the individual leader manages the challenging transition into a new leadership context and how their approach to leadership facilitates or inhibits successful transition (Avolio, 2010). Leader assimilation programs have been developed to assimilate new leaders (Manderscheid, 2008); however, assimilation is not necessarily a desired organisational outcome (Denis and Pineault, 2000). In this paper we critically review the limited literature on inter-sector leader transitions and transformational change outcomes and argue for a mutual accommodation approach. We draw on our own initial empirical work to propose the elements of a program for achieving this outcome from the perspective of the public organisation and the inter-sector appointee.

Consumers’ intentions to adopt 3G in regional cities in China : a comparative study

Rapid mobile technological evolution and the large economic stake in commercial development of mobile technological innovation make it necessary to understand consumers' motivations towards the latest advanced and updated technologies and services. 3G (the third generation of mobile communication technology) recently started its commercial development in the world‘s largest mobile communication market, China, after being delayed for a few years. Although China fell behind in commercially developing 3G, it is difficult to ignore studying this area, given the size of the market and promising future developments. This market deserves focused research attention, especially in terms of consumer behaviour towards the adoption of mobile technological innovation. Thus, the program of research in this thesis was designed to investigate how Chinese consumers respond to the use of this newly launched mobile technological innovation, with a focus on what factors affect their 3G adoption intentions. It aimed to yield important insights into Chinese consumers‘ innovation adoption behaviours and to contribute to marketing and innovation adoption research. Furthermore, it has been documented that Chinese consumers vary widely between regions in dialect, lifestyle, culture, purchasing power and consumption attitudes. Based on economic development and local culture, China can be divided geographically into distinctive regional consumer markets. Consequently, the results of consumer behaviour research in one region may not necessarily be extrapolated to other regions. In order to better understand Chinese consumers, the disparities between regions should not be overlooked. Therefore, another objective of this program of research was to examine regional variances in consumers' innovation adoption, specifically to identify the similarities and differences in factors influencing 3G adoption, contributing to intra-cultural studies. An extensive literature review identified two gaps: current China-based innovation adoption research studies are limited in providing adequate prediction and explanation of Chinese consumers' intentions to adopt 3G; and there was limited knowledge about the differences between regional Chinese consumers in innovation adoption. Two research questions therefore were developed to address these gaps: 1) What factors influence Chinese consumers' intentions to adopt 3G? 2) How do Chinese consumers differ between regional markets in the relative influence of the factors in determining their intentions to adopt 3G? In accordance with postpositivist research philosophy, two studies were designed to answer the research questions, using mixed methods. To meet the research objectives, the two studies were both conducted in three regional cities, namely Beijing, Shanghai and Wuhan, centred in the three regions of North China, East China and Central China respectively, with sufficient cultural and economical regional variances. Study One was an exploratory study with qualitative research methods. It involved 45 in-depth interviews in the three research cities to gain rich insights into the research context from natural settings. Eight important concepts related to 3G adoption were generated from analysis of the interview data, namely utilitarian expectation, hedonic expectation, status gains, status loss avoidance, normative influence, external influence, cost and quality concern. The concepts of social loss avoidance and quality concern were two unique findings, whereas the other concepts were similar to the findings in Western innovation adoption studies. Moreover, variances in 3G adoption between three groups of regional consumers were also identified, focusing on the perceptions of two concepts, namely status gains and normative influence. The conceptual research model was then developed incorporating the eight concepts plus the dependent variable of adoption intention. The hypothesized relationships between the nine constructs and hypotheses about the differences between regional consumers in 3G adoption were informed by the findings of Study One and the literature reviewed. Study Two was a quantitative study involving a web-based survey and statistical analysis procedure. The web-based survey attracted 800 residents from the three research cities, 270 from Beijing, 265 from Shanghai and 265 from Wuhan. They comprised three research samples for this study and consequently three sets of data were obtained. The data was analysed by Structural Equation Modelling together with Multi-group Analysis. The analysis confirmed that the concepts generated in Study One were influential factors affecting Chinese consumers' 3G adoption intention, with the exception of the concept external influence. Differences were found between the samples in the three research cities in the effect of hedonic expectation, status gains, status loss avoidance and normative influence on 3G adoption intention. The two Studies undertaken in this thesis contributed a better understanding of Chinese consumers' intentions to adopt advanced mobile technological innovation, namely 3G, in three regional markets. This knowledge contributes to innovation adoption and intra-cultural research, as well as consumer behaviour theory. It is also able to inform international and domestic telecommunication companies to develop and deliver more effective marketing strategies across Chinese regional markets. Limitations in the research were identified in terms of the sampling techniques used and the design of the two Studies. Future research was suggested in other Chinese regional markets and into consumer adoption of other types of mobile technological innovations.

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.