Expression pattern of Oct-4, Sox2, and c-Myc in the primary culture of human dental pulp derived cells

Dental pulp cells (DPCs) have shown promising potential in dental tissue repair and regeneration. However, during in vitro culture, these cells undergo replicative senescence and result in significant alteration in cell proliferation and differentiation. Recently, the transcription factors of Oct-4, Sox2, c-Myc, and Klf4 have been reported to play a regulatory role in the stem cell self-renewal process, namely cell reprogramming. Therefore, it is interesting to know whether the replicative senescence during the culture of dental pulp cells is related to the diminishing of the expression of these transcription factors. In this study, we investigated the expression of the reprogramming markers Oct-4, Sox2, and c-Myc in the in vitro explant cultured dental pulp tissues and explant cultured dental pulp cells (DPCs) at various passages by immunofluorescence staining and real-time polymerase chain reaction analysis. Our results demonstrated that Oct-4, Sox2, and c-Myc translocated from nucleus in the first 2 passages to cytoplasm after the third passage in explant cultured DPCs. The mRNA expression of Oct-4, Sox2, and c-Myc elevated significantly over the first 2 passages, peaked at second passage (P < .05), and then decreased along the number of passages afterwards (P < .05). For the first time we demonstrated that the expression of reprogramming markers Oct-4, Sox2, and c-Myc was detectable in the early passaged DPCs, and the sequential loss of these markers in the nucleus during DPC cultures might be related to the cell fate of dental pulp derived cells during the long-term in vitro cultivation under current culture conditions.

Zonal chondrocyte subpopulations reacquire zone-specific characteristics during in Vitro redifferentiation

Background: If chondrocytes from the superficial, middle, and deep zones of articular cartilage could maintain or regain their characteristic properties during in vitro culture, it would be feasible to create constructs comprising these distinctive zones. ----- ----- Hypothesis: Zone-specific characteristics of zonal cell populations will disappear during 2-dimensional expansion but will reappear after 3-dimensional redifferentiation, independent of the culture technique used (alginate beads versus pellet culture).----- ----- Study Design: Controlled laboratory study.----- ----- Methods: Equine articular chondrocytes from the 3 zones were expanded in monolayer culture (8 donors) and subsequently redifferentiated in pellet and alginate bead cultures for up to 4 weeks. Glycosaminoglycans and DNA were quantified, along with immunohistochemical assessment of the expression of various zonal markers, including cartilage oligomeric protein (marking cells from the deeper zones) and clusterin (specifically expressed by superficial chondrocytes).----- ----- Results: Cell yield varied between zones, but proliferation rates did not show significant differences. Expression of all evaluated zonal markers was lost during expansion. Compared to the alginate bead cultures, pellet cultures showed a higher amount of glycosaminoglycans produced per DNA after redifferentiation. In contrast to cells in pellet cultures, cells in alginate beads regained zonal differences, as evidenced by zone-specific reappearance of cartilage oligomeric protein and clusterin, as well as significantly higher glycosaminoglycans production by cells from the deep zone compared to the superficial zone.----- ----- Conclusion: Chondrocytes isolated from the 3 zones of equine cartilage can restore their zone-specific matrix expression when cultured in alginate after in vitro expansion.

Information security management : a case study of an information security culture

This thesis argues that in order to establish a sound information security culture it is necessary to look at organisation's information security systems in a socio- technical context. The motivation for this research stems from the continuing concern of ineffective information security in organisations, leading to potentially significant monetary losses. It is important to address both technical and non- technical aspects when dealing with information security management. Culture has been identified as an underlying determinant of individuals' behaviour and this extends to information security culture, particularly in developing countries. This research investigates information security culture in the Saudi Arabia context. The theoretical foundation for the study is based on organisational and national culture theories. A conceptual framework for this study was constructed based on Peterson and Smith's (1997) model of national culture. This framework guides the study of national, organisational and technological values and their relationships to the development of information security culture. Further, the study seeks to better understand how these values might affect the development and deployment of an organisation's information security culture. Drawing on evidence from three exploratory case studies, an emergent conceptual framework was developed from the traditional human behaviour and the social environment perspectives used in social work, This framework contributes to in- formation security management by identifying behaviours related to four modes of information security practice. These modes provide a sound basis that can be used to evaluate individual organisational members' behaviour and the adequacy of ex- isting security measures. The results confirm the plausibility of the four modes of practice. Furthermore, a final framework was developed by integrating the four modes framework into the research framework. The outcomes of the three case stud- ies demonstrate that some of the national, organisational and technological values have clear impacts on the development and deployment of organisations' informa- tion security culture. This research, by providing an understanding the in uence of national, organi- sational and technological values on individuals' information security behaviour, contributes to building a theory of information security culture development within an organisational context. The research reports on the development of an inte- grated information security culture model that highlights recommendations for developing an information security culture. The research framework, introduced by this research, is put forward as a robust starting point for further related work in this area.

Culture and creative industries in Australia

This paper draws upon the Australian case to argue that the case for support for cultural production and cultural infrastructure has been strengthened overall by its alignment to economic policy goals. In this respect, the rise of creative industries policy discourses is consistent with trends in thinking about cultural policy that have their roots in the Creative Nation strategies of the early 1990s. In terms of the earlier discussion, cultural policy is as much driven by Schumpeterian principals as it is by Keynesian ones. Such an approach is not without attendant risks, and two stand out. The first is the risk of marginalizing the arts, through a policy framework that gives priority to developing the digital content industries, and viewing the creative industries as primarily an innovation platform. The second is that other trends in the economy, such as the strong Australian dollar resulting from the mining boom, undercuts the development of cultural production in the sections of the creative industries where international trade and investment is most significant, such as the film industry and computer games. Nonetheless, after over a decade of vibrant debate, this focus on linking the cultural and economic policy goals of the creative industries has come to be consistent with broader international trends in the field.

Supply chain sustainability : a relationship management approach moderated by culture and commitment

This research explores the nature of relationship management on construction projects in Australia and examines the effects of culture, by means of Schwarz’s value survey, on relationships under different contract strategies. The research was based on the view that the development of a sustainable supply chain depends on the transfer of knowledge and capabilities from the larger players in the supply chain through collaboration brought about by relationship management. The research adopted a triangulated approach in which quantitative data were collected by questionnaire, interviews were conducted to explore and enrich the quantitative data and case studies were undertaken in order to illustrate and validate the findings. The aim was to investigate how values and attitudes enhance or reduce the incorporation of the supply chain into the project. From the research it was found that the degree of match and mismatch between values and contract strategy impacts commitment and the engagement and empowerment of the supply chain.

The validity and structure of culture-level personality scores: Data from ratings of young adolescents

We examined properties of culture-level personality traits in ratings of targets (N=5,109) ages 12 to 17 in 24 cultures. Aggregate scores were generalizable across gender, age, and relationship groups and showed convergence with culture-level scores from previous studies of self-reports and observer ratings of adults, but they were unrelated to national character stereotypes. Trait profiles also showed cross-study agreement within most cultures, 8 of which had not previously been studied. Multidimensional scaling showed that Western and non-Western cultures clustered along a dimension related to Extraversion. A culture-level factor analysis replicated earlier findings of a broad Extraversion factor but generally resembled the factor structure found in individuals. Continued analysis of aggregate personality scores is warranted.

Review of refugee mental health interventions following resettlement : best practices and recommendations

There are increasing numbers of refugees worldwide, with approximately 16 million refugees in 2007 and over 2.5 million refugees resettled in the United States since the start of its humanitarian program. Psychologists and other health professionals who deliver mental health services for individuals from refugee backgrounds need to have confidence that the therapeutic interventions they employ are appropriate and effective for the clients with whom they work. The current review briefly surveys refugee research, examines empirical evaluations of therapeutic interventions in resettlement contexts, and provides recommendations for best practices and future directions in resettlement countries. The resettlement interventions found to be most effective typically target culturally homogeneous client samples and demonstrate moderate to large outcome effects on aspects of traumatic stress and anxiety reduction. Further evaluations of the array of psychotherapeutic, psychosocial, pharmacological, and other therapeutic approaches, including psychoeducational and community-based interventions that facilitate personal and community growth and change, are encouraged. There is a need for increased awareness, training and funding to implement longitudinal interventions that work collaboratively with clients from refugee backgrounds through the stages of resettlement.

Building organizational capability through a quality assurance program within a business school and encouraging organizational culture change

This study explores organizational capability and culture change through a project developing an assurance of learning program in a business school. In order to compete internationally for high quality faculty, students, strategic partnerships and research collaborations it is essential for Universities to develop and maintain an international focus and a quality produce that predicts excellence in the student experience and graduate outcomes that meet industry needs. Developing, marketing and delivering that quality product requires an organizational strategy to which all members of the organization contribute and adhere. Now, the ability to acquire, share and utilize knowledge has become a critical organizational capability in academia as well as other industries. Traditionally the functional approach to business school structures and disparate nature of the social networks and work contact limit the sharing of knowledge between academics working in different disciplines. In this project a community of practice program was established to include academics in the development of an embedded assurance of learning program affecting more than 5000 undergraduate students and 250 academics from nine different disciplines across four schools. The primary outcome from the fully developed and implemented assurance of learning program was the five year accreditation of the business schools programs by two international accrediting bodies, EQUIS and AACSB. However this study explores a different outcome, namely the change in organizational culture and individual capabilities as academics worked together in teaching and learning teams. This study uses a survey and interviews with academics involved, through a retrospective panel design which contained an experimental group and a control group. Results offer insights into communities of practice as a means of addressing organizational capability and changes in organizational culture. Knowledge management and shared learning can achieve strategic and operational benefits equally within academia as within other industrial enterprises but it comes at a cost. Traditional structures, academics that act like individual contractors and deep divides across research, teaching and service interest served a different master and required fewer resources. Collaborative structures; fewer master categories of discrete knowledge areas; specific strategic goals; greater links between academics and industry; and the means to share learned insights will require a different approach to resourcing both the individual and the team.

Enhancing business continuity management capacities in the insurance industry : a Southeast Asian perspective

As a resilience enhancing practice, business continuity management (BCM) can play an important role in aiding preparation of the insurance industry for coping with the losses incurred by major discontinuity incidents: regardless of cause. Acknowledging the increasing frequency of unpredictable man-made disasters and natural catastrophes, the insurance industry would benefit from examining and implementing, where suitable, key elements of BCM. Such strategic decisions would assist insurers and re-insurers collectively to enhance mutual capability to respond to, and recover from, the impact of significant losses. This paper presents a comparison of opinions about BCM practitioners in both retail and re-insurance companies on the importance of generic continuity practices with actual levels of BCM practice across the two industry groups in Southeast Asia. It suggests means by which multi-lateral cooperation across Asian economies and between retail and re-insurance market segments might enhance the viability of the insurance industry in the face of increased stress from major natural and socio-technical hazards.

Safety leaders' perceptions of safety culture in a large Australian construction organisation

Safety culture in the construction industry is a growing research area. The unique nature of construction industry works – being project-based, varying in size and focus, and relying on a highly transient subcontractor workforce – means that safety culture initiatives cannot be easily translated from other industries. This paper reports on the first study in a three year collaborative industry and university research project focusing on safety culture practices and development in one of Australia’s largest global construction organisations. The first round of a modified Delphi method is reported, and describes the insights gained from 41 safety leaders’ perceptions and understandings of safety culture within the organisation. In-depth, semi-structured interviews were conducted, and will be followed by a quantitative perception survey with the same sample. Participants included Senior Executives, Corporate Managers, Project Managers, Safety Managers and Site Supervisors. Leaders’ definitions and descriptions of safety culture were primarily action-oriented and some confusion was evident due to the sometimes implicit nature of culture in organisations. Leadership was identified as a key factor for positive safety culture in the organisation, and there was an emphasis on leaders demonstrating commitment to safety, and being visible to the project-based workforce. Barriers to safety culture improvement were also identified, with managers raising diverse issues such as the transient subcontractor workforce and the challenge of maintaining safety as a priority in the absence of safety incidents, under high production pressures. This research is unique in that it derived safety culture descriptions from key stakeholders within the organisation, as opposed to imposing traditional conceptualisations of safety culture that are not customised for the organisation or the construction industry more broadly. This study forms the foundation for integrating safety culture theory and practice in the construction industry, and will be extended upon in future studies within the research program.

Independent cinema in the Chinese film industry

Chinese independent cinema has developed for more than twenty years. Two sorts of independent cinema exist in China. One is underground cinema, which is produced without official approvals and cannot be circulated in China, and the other are the films which are legally produced by small private film companies and circulated in the domestic film market. This sort of ‘within-system’ independent cinema has played a significant role in the development of Chinese cinema in terms of culture, economics and ideology. In contrast to the amount of comment on underground filmmaking in China, the significance of ‘within-system’ independent cinema has been underestimated by most scholars. This thesis is a study of how political management has determined the development of Chinese independent cinema and how Chinese independent cinema has developed during its various historical trajectories. This study takes media economics as the research approach, and its major methods utilise archive analysis and interviews. The thesis begins with a general review of the definition and business of American independent cinema. Then, after a literature review of Chinese independent cinema, it identifies significant gaps in previous studies and reviews issues of traditional definition and suggests a new definition. After several case studies on the changes in the most famous Chinese directors’ careers, the thesis shows that state studios and private film companies are two essential domestic backers for filmmaking in China. After that, the body of the thesis provides an examination of the development of ‘within-system’ independent cinema. Specifically, three factors: government intervention, the majors’ performance (state studios and, later, the conglomerates) and the market conduct of independent cinema at various points in their trajectories are studied. The key findings of the study are as follows: First, most scholars have overlooked the existence and the significance of within-system Chinese independent cinema. Drawing on an American definition of the independent sector, this thesis proposes a definition of the sector in China: namely, any film that has not been financed, produced, and/or distributed by majors. The thesis also notes important contradictions in applying this definition: i.e. film-making is still dependent on policies that frame industry development. The thesis recognises that major tensions apply to filmmaking in China, which significantly differentiates the Chinese independents from those in the US. Second, the development of Chinese independent cinema is the result the rise of the private sector and the decline of the state studio system. As state studios encountered difficulties the private sector moved forward; consequently the environment improved for independent cinema. Third, before 2003, the film industry in China had little commercialisation. The government controlled independent cinema by means of license and censorship. State studios produced main melody films and Hollywood attracted most of the audiences. Many independent filmmakers focused on commercial films, thus contributing to film commercialisation. Fourth, after 2003, the film industry became increasingly fragmented. The government created distribution and exhibition opportunities for main melody films; conglomerates collaborated with Hong Kong players; Hong Kong co-productions and Hollywood occupied the film market; and small private film companies produced main melody films in order to earn meagre profits. The original contribution of the thesis is to advance the study of Chinese independent cinema. The study suggests a reasonable and practical definition of Chinese independent cinema. It shows how the Chinese government authorities have implemented economic measures to gain ideological control in the film industry. Finally, this the first study on Chinese independent cinema applying a synthesis of economic, political and historical perspectives.

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.