The active site residue V266 of Chlamydial HtrA is critical for substrate binding during both in vitro and in vivo conditions

HtrA is a complex, multimeric chaperone and serine protease important for the virulence and survival of many bacteria. Chlamydia trachomatis is an obligate, intracellular bacterial pathogen that is responsible for severe disease pathology. C. trachomatis HtrA (CtHtrA) has been shown to be highly expressed in laboratory models of disease. In this study, molecular modelling of CtHtrA protein active site structure identified putative S1-S3 subsite residues I242, I265, and V266. These residues were altered by site-directed mutagenesis, and these changes were shown to considerably reduce protease activity on known substrates and resulted in a narrower and distinct range of substrates compared to wild type. Bacterial two-hybrid analysis revealed that CtHtrA is able to interact in vivo with a broad range of protein sequences with high affinity. Notably, however, the interaction was significantly altered in 35 out of 69 clones when residue V266 was mutated, indicating that this residue has an important function during substrate binding.

Organisational culture in airworthiness management programs : developing a measurement model

All civil and private aircraft are required to comply with the airworthiness standards set by their national airworthiness authority and throughout their operational life must be in a condition of safe operation. Aviation accident data shows that over twenty percent of all fatal accidents in aviation are due to airworthiness issues, specifically aircraft mechanical failures. Ultimately it is the responsibility of each registered operator to ensure that their aircraft remain in a condition of safe operation, and this is done through both effective management of airworthiness activities and the effective program governance of safety outcomes. Typically, the projects within these airworthiness management programs are focused on acquiring, modifying and maintaining the aircraft as a capability supporting the business. Program governance provides the structure through which the goals and objectives of airworthiness programs are set along with the means of attaining them. Whilst the principal causes of failures in many programs can be traced to inadequate program governance, many of the failures in large scale projects can have their root causes in the organisational culture and more specifically in the organisational processes related to decision-making. This paper examines the primary theme of project and program based enterprises, and introduces a model for measuring organisational culture in airworthiness management programs using measures drawn from 211 respondents in Australian airline programs. The paper describes the theoretical perspectives applied to modifying an original model to specifically focus it on measuring the organisational culture of programs for managing airworthiness; identifying the most important factors needed to explain the relationship between the measures collected, and providing a description of the nature of these factors. The paper concludes by identifying a model that best describes the organisational culture data collected from seven airworthiness management programs.

Characterization of in vitro Chlamydia muridarum persistence and utilization in an in vivo mouse model of Chlamydia vaccine

Problem: Chlamydia trachomatis genital tract infections are easily treated with antibiotics, however the majority of infections are asymptomatic and therefore untreated, highlighting the need for a vaccine. Because most infections are asymptomatic, vaccination could potentially be administered to individuals who may have an acute infection at that time. In such individuals the effect of vaccination on the existing infection is unknown; however one potential outcome could be the development of a persistent infection. In vitro chlamydial persistence has been well characterized in various strains, however there have been no reported studies in C. muridarum. Method of Study: We performed ultrastructural characterization, and transcriptome analysis of selected genes. We then used the transcriptional profiles of the selected genes to examine whether intranasal immunization of mice during an active genital infection would induce persistence in the upper reproductive tract of female mice. Results and Conclusions: We found that persistence developed in the oviducts of mice as a result of immunization. This is a significant finding, not only because it is the first time that C. muridarum persistence has been characterized in vitro, but also due to the fact that there is minimal characterization of in vivo persistence of any chlamydial species. This highlights the importance of the timing of vaccination in individuals.

The International Film and Television School in Cuba : for a stronger media culture in the global south

A little-known facet of Cuban internationalism is the Cuba shares in the education of young people who want to help build a stronger media culture that represents voices from the global South. Cuba was instrumental in the establishment and operation of the International Film and Television School at San Antonio de los Baños. The Cuban government provided the location and buildings for the school, and among the range of international media professionals who teach the students are selected Cuban professors from the Institute of the Arts, based n Havana. The International Film and Television School is supported by funding from Spain and other countries, and by the willingness of international media professionals to teach short courses for little more than an honorarium. Cuba used to provide full scholarships for student from the South to study a two-year course in film or television, but now charges fees for its three-year diploma course.

Consuming Urban Culture in Contemporary Vietnam

Vietnam is currently undergoing a metamorphosis from a relatively closed society with a centrally planned economy, to a rapidly urbanising one with a global outlook. These changes have been the catalyst for an exciting ferment of activity in popular culture. This volume contains contributions from scholars engaged in the most up-to-date social research in Vietnam, as well as some of Vietnam's most popular cultural producers who are forging new ways of imagining the present whilst at the same time engaging actively in reinterpreting the past. The diverse ways that Vietnam is culturally and socially negotiating the future are examined as the book addresses issues of indigenisation of cultural influences, ambivalence surrounding change, and the consistent blurring of boundaries between informal, non-state cultural activities and formal institutional structures in the evolution of a civil society in Vietnam.

Organizational culture in airworthiness management programmes : extending an existing measurement model

All civil and private aircraft are required to comply with the airworthiness standards set by their national airworthiness authority and throughout their operational life must be in a condition of safe operation. Aviation accident data shows that over 20% of all fatal accidents in aviation are due to airworthiness issues, specifically aircraft mechanical failures. Ultimately it is the responsibility of each registered operator to ensure that their aircraft remain in a condition of safe operation, and this is done through both effective management of airworthiness activities and the effective programme governance of safety outcomes. Typically, the projects within these airworthiness management programmes are focused on acquiring, modifying and maintaining the aircraft as a capability supporting the business. Programme governance provides the structure through which the goals and objectives of airworthiness programmes are set along with the means of attaining them. Whilst the principal causes of failures in many programmes can be traced to inadequate programme governance, many of the failures in large-scale projects can have their root causes in the organizational culture and more specifically in the organizational processes related to decision-making. This paper examines the primary theme of project and programme-based enterprises, and introduces a model for measuring organizational culture in airworthiness management programmes using measures drawn from 211 respondents in Australian airline programmes. The paper describes the theoretical perspectives applied to modifying an original model to specifically focus it on measuring the organizational culture of programmes for managing airworthiness; identifying the most important factors needed to explain the relationship between the measures collected, and providing a description of the nature of these factors. The paper concludes by identifying a model that best describes the organizational culture data collected from seven airworthiness management programmes.

Safety culture in supply chains : 'Customer is king'

Despite the presence of many regulations governing the operation of heavy vehicles and supply chains in Australia, the truck driving sector continues to have the highest incidence of fatal injuries compared to all other industries. The working environment has been the focus of attention by safety researchers during the past few decades, with particular consideration been given to the concept ‘safety culture’ and how to maintain, modify and advance responses to occupational risk. One important aspect of the heavy industry which sets it apart is the existence of cultural or sub-cultural influences at an industry wide and occupation-specific level rather than organisational level. This paper reports on the findings of stakeholder’s perceptions of the influences of power and control, and culture on industry safety. In-depth structured interviews were conducted during 2011 with Australian industry stakeholders (n=31). The questioning surrounded decision-making processes with regards to identifying risks, self-monitoring and reducing risky activities; as well as how power-affected relationships may influence the operational performance of supply chains and impacts on driver safety. One of the most significant findings from these interviews relates to the notion of power. The perception that the ‘Customer is King’ was widely viewed, with the majority of stakeholders believing that there exists a ‘master slave mentality’ in the industry. There appears to be great frustration in the industry as to the apparent immunity of customers (particularly retail supply chains) to their responsibilities. There was also a strong perception that the customer holds the balance of power by covertly employing remuneration-related incentives and pressures. Smaller trucking companies are perceived as being more vulnerable to the pressure of customer expectations.

In vitro models for investigating keratinocyte responses to ultraviolet B radiation

This thesis describes the use of 2- and 3-dimensional cell-based models for studying how skin cells respond to ultraviolet radiation. These methods were used to investigate skin damage and repair after exposure to radiation in the context of skin cancer development. Interactions between different skin cell types were demonstrated as being significant in protecting against ultraviolet radiation-induced skin damage. This has important implications in understanding how skin cancers occur, as well as in the development of new strategies to prevent and treat them.

Integration of Saudi Arabia’s conservative Islamic culture in sustainable housing design

The cities of Saudi Arabia have perhaps the largest growth rates of cities in the Middle East, such that it has become a cause in shortage of housing for mid and low-income families, as is the case in other developing countries. Even when housing is found, it is not sustainable nor is it providing the cultural needs of those families. The aim of this paper is to integrate the unique conservative Islamic Saudi culture into the design of sustainable housing. This paper is part of a preliminary study of an on-going PhD thesis, which utilises a semistructured interview of a panel of nine experts in collecting the data. The interviews consisted of ten questions ranging from general questions such as stating their expertise and work position to more specific question such as listing the critical success factors and/or barriers for applying sustainability to housing in Saudi Arabia. Since the participants were selected according to their experience, the answers to the interview questions were satisfactory where the generation of the survey questions for the second stage in the PhD thesis took place after analysing the participant’s answers to the interview questions. This paper recommends design requirements for accommodating the conservative Islamic Saudi Culture in low cost sustainable houses. Such requirements include achieving privacy through the use of various types of traditional Saudi architectural elements, such as the method of decorative screening of windows, called Mashrabiya, and having an inner courtyard where the house looks inward rather than outward. Other requirements include educating firms on how to design sustainable housing, educating the public on the advantages of sustainable housing and implementing new laws that enforce the utilisation of sustainable methods to housing construction. This paper contributes towards the body of knowledge by proposing initial findings on how to integrate the conservative Islamic culture of Saudi Arabia into the design of a sustainable house specifically for mid and low-income families. This contribution can be implemented on developing countries in the region that are faced with housing shortage for mid and low-income families.

The impact of organisational culture in project selection process : case of public infrastructure project in Indonesia

The thesis develops an Organisational Culture (OC) based framework for potentially improving the current practice of the infrastructure project selection process (PSP) in Indonesia, particularly for regional road projects. This framework is developed to address the suggestion emanating from the research that there is a need for strengthening the relevant organisational culture dimensions and making changes to current organisational culture profiles to support the effectiveness of the decision-making process of the current PSP. The findings not only benefit existing knowledge but also provide practical contributions for those decision-makers within the Indonesian regional government that are involved in PSP, i.e. by raising the awareness of the influence and the role of organisational culture.

In vitro functional characterisation of IGF-I : VN-induced breast cancer progression

Members of the insulin-like growth factor (IGF) family have been shown to play critical roles in normal growth and development, as well as in tumour biology. The IGF system is complex and the biological effects of the IGFs are determined by their diverse interactions between many molecules, including their interactions with extracellular matrix (ECM) proteins. Recent studies have demonstrated that IGFs associate with the ECM protein vitronectin (VN) through IGF-binding proteins (IGFBP) and that this interaction modulates IGF-stimulated biological functions, namely cell migration and cell survival through the cooperative involvement of the type-I IGF receptor (IGF-1R) and VN-binding integrins. Since IGFs play important roles in the transformation and progression of breast cancer and VN has been found to be over-expressed at the leading edge of breast tumours, this project aimed to describe the effects of IGF-I:VN interactions on breast cell function. This was undertaken to dissect the molecular mechanisms underlying IGF-I:VN-induced responses and to design inhibitors to block the effects of such interactions. The studies described herein demonstrate that the increase in migration of MCF-7 breast cancer cells in response to the IGF-I:IGFBP-5:VN complex is accompanied by differential expression of genes known to be involved in migration, invasion and/or survival, including Tissue-factor (TF), Stratifin (SFN), Ephrin-B2, Sharp-2 and PAI-1. This „migration gene signature‟ was confirmed using real-time PCR analysis. Substitution of the native IGF-I within the IGF-I:IGFBP:VN complex with the IGF-I analogue, \[L24]\[A31]-IGF-I, which has a reduced affinity for the IGF-1R, failed to stimulate cell migration and interestingly, also failed to induce the differential gene expression. This supports the involvement of the IGF-1R in mediating these changes in gene expression. Furthermore, lentiviral shRNA-mediated stable knockdown of TF and SFN completely abrogated the increased cell migration induced by IGF-I:IGFBP:VN complexes in MCF-7 cells. Indeed, when these cells were grown in 3D Matrigel™ cultures a decrease in the overall size of the 3D spheroids in response to the IGF-I:IGFBP:VN complexes was observed compared to the parental MCF-7 cells. This suggests that TF and SFN have a role in complex-stimulated cell survival. Moreover, signalling studies performed on cells with the reduced expression of either TF or SFN had a decreased IGF-1R activation, suggesting the involvement of signalling pathways downstream of IGF-1R in TF- and/or SFN-mediated cell migration and cell survival. Taken together, these studies provide evidence for a common mechanism activated downstream of the IGF-1R that induces the expression of the „migration gene signature‟ in response to the IGF-I:IGFBP:VN complex that confers breast cancer cells the propensity to migrate and survive. Given the functional significance of the interdependence of ECM and growth factor (GF) interactions in stimulating processes key to breast cancer progression, this project aimed at developing strategies to prevent such growth factor:ECM interactions in an effort to inhibit the downstream functional effects. This may result in the reduction in the levels of ECM-bound IGF-I present in close proximity to the cells, thereby leading to a reduction in the stimulation of IGF-1R present on the cell surface. Indeed, the inhibition of IGF-I-mediated effects through the disruption of its association with ECM would not alter the physiological levels of IGF-I and potentially only exert effects in situations where abnormal over expression of ECM proteins are found; namely carcinomas and hyperproliferative diseases. In summary, this PhD project has identified novel, innovative and realistic strategies that can be used in vitro to inhibit the functions exerted by the IGF-I:IGFBP:VN multiprotein complexes critical for cancer progression, with a potential to be translated into in vivo investigations. Furthermore, TF and SFN were found to mediate IGF-I:IGFBP:VN-induced effects, thereby revealing their potential to be used as therapeutic targets or as predictive biomarkers for the efficacy of IGF-1R targeting therapies in breast cancer patients. In addition to its therapeutic and clinical scope, this PhD project has significantly contributed to the understanding of the role of the IGF system in breast tumour biology by providing valuable new information on the mechanistic events underpinning IGF-I:VN-mediated effects on breast cell functions. Furthermore, this is the first instance where favourable binding sites for IGF-II, IGFBP-3 and IGFBP-5 on VN have been identified. Taken together, this study has functionally characterised the interactions between IGF-I and VN and through innovative strategies has provided a platform for the development of novel therapies targeting these interactions and their downstream effects.

Influence of osteocytes in the in vitro and in vivo β-tricalcium phosphate-stimulated osteogenesis

Osteocytes, known to act as the main regulators of bone homeostasis, have become a major focus in the field of bone research. Bioactive ceramics have been widely used for bone regeneration. However, there are few studies about the interaction of osteocytes with bioceramics. The effects of osteocytes on the in vitro and in vivo osteogenesis of bioceramics are also unclear. The aim of this study was to investigate the role of osteocytes on the b-tricalcium phosphate (b-TCP) stimulated osteogenesis. It was found that osteocytes responded to the b-TCP stimulation, leading to the release of Wnt (wingless-related MMTV integration site), which enhanced osteogenic differentiation of bone marrow stromal cells via Wnt signaling pathway. Receptor activator of nuclear factor kappa B ligand, an osteoclast inducer, was also upregulated, indicating that osteocytes would also participated in activation of osteoclasts, which played a major role in the degradation process of b-TCP and new bone remodeling. In vivo studies further demonstrated that when the material was completely embedded by newly formed bone, the only cell contacting with the material was osteocyte. However, the material would eventually be degraded and replaced by the new bone, requiring the participation of osteoclasts and osteoblasts, which were demonstrated by using immunostaining in this study. As the only cell contacting with the material, osteocytes probably acted in a regulatory role to regulate the surrounding osteoclasts and osteoblasts. Osteocytes were also found to participate in the maturation of osteoblasts and the mineralization process of biomaterials, by upregulating E11 (podoplanin) and dentin matrix protein 1 expression. These findings indicated that osteocytes involved in bone biomaterial-mediated osteogenesis and biomaterial degradation, providing valuable insights into the mechanism of material-stimulated osteogenesis, and a novel strategy to optimize the evaluating system for the biological properties of biomaterials.

Culture in and culture of health promotion: tensions at the interface of Australian Indigenous health promotion

With persisting health inequalities across and between diverse populations, health promotion must consider its engagement with the culture concept in achieving better health for all. By way of a conversation between an Indigenous and non-Indigenous health promotion practitioner, this unique presentation will critically examine the cultural practice of health promotion for Indigenous Australians. Culture becomes the central tenant of this conversation – but not culture in the sense of something to “fix” to improve Indigenous health, or import to make mainstream practices “culturally appropriate”. Rather, the somewhat invisible culture of Australian health promotion practice itself is highlighted. The enthusiasm of mainstream health promotion practice for risk and reductionism supplants biological determinism with a cultural determinism that constructs culture as illness-producing. This is in contrast to Indigenous perspectives of culture in which it is described as integral to individual and community health and well-being. Whilst empowerment features strongly within global health promotion discourses, the preoccupation of health promotion with the inherent deficit/behavioural change approach is an all too convenient distraction from the broader structural factors impacting on the health of Indigenous Australians. That Indigenous Australians have not benefitted from successful public health policy interventions in the same way as the general population is in itself revealing of the culture of health promotion practice in Australia and it is somewhat ironic that the health promotion fraternity seems not to have questioned its own practice. This conversation aims to encourage health promotion practitioners, researchers and policy makers to interrogate the cultural assumptions of their own practice and of the public health system they are part of and consider how to embed and empower the voices and experiences of those who are ‘culturally othered’ within health promotion practice.

Modulation of in vitro platelet 5-HT release by species of Erythrina and Cymbopogon

Extracts of Australian plants were screened to detect constituents affecting adenosine di-phosphate (ADP) induced platelet aggregation and [14C]5-hydroxytryptamine (5-HT) release. Extracts of four tested plants including, Eremophila gilesii, Erythrina vespertilio, Cymbopogon ambiguus, and Santalum acuminatum, were found to cause significant inhibition of platelet 5-HT release. Inhibition levels ranged from 56-98%, and was not due to the non-specific effects of protein binding tannins. These extracts, and those we have previously identified as being active, were examined further to determine if they affect epinephrine (EPN), arachidonic acid (A.A) or collagen stimulated platelet aggregation and 5-HT release. Among those extracts investigated, we found that both the methanolic extract of E. vespertilio and the dichloromethane (DCM) extract of C. ambiguus were most potent and caused significant inhibition of platelet activation induced by EPN, A.A and to a lesser extent by collagen. Inhibition of ADP induced platelet 5-HT release by both of these extracts, was dose-dependent, with IC50 values for E. vespertilio and C. ambiguus estimated to be 20.4 microl (1.855 mg/ml) and 8.34 microl (0.758 mg/ml), respectively. Overall, C. ambiguus exhibited most activity and also caused dose-dependent inhibition of A.A induced platelet activation. These results indicate that inhibition may occur specifically at a site within the A.A pathway, and suggest the presence of a cyclo-oxygenase inhibitor. Both E. vespertilio and C. ambiguus are reported to be traditional headache treatments, with the present study providing evidence that they affect 5-HT release.