Pathogenicity of Plesiomonas shigelloides : interactions with eukaryotic host cells in vitro

Diarrhoea is one of the leading causes of morbidity and mortality in populations in developing countries and is a significant health issue throughout the world. Despite the frequency and the severity of the diarrhoeal disease, mechanisms of pathogenesis for many of the causative agents have been poorly characterised. Although implicated in a number of intestinal and extra-intestinal infections in humans, Plesiomonas shigelloides generally has been dismissed as an enteropathogen due to the lack of clearly demonstrated virulence-associated properties such as production of cytotoxins and enterotoxins or invasive abilities. However, evidence from a number of sources has indicated that this species may be the cause of a number of clinical infections. The work described in this thesis seeks to resolve this discrepancy by investigating the pathogenic potential of P. shigelloides using in vitro cell models. The focus of this research centres on how this organism interacts with human host cells in an experimental model. Very little is known about the pathogenic potential of P. shigel/oides and its mechanisms in human infections and disease. However, disease manifestations mimic those of other related microorganisms. Chapter 2 reviews microbial pathogenesis in general, with an emphasis on understanding the mechanisms resulting from infection with bacterial pathogens and the alterations in host cell biology. In addition, this review analyses the pathogenic status of a poorly-defined enteropathogen, P. shigelloides. Key stages of pathogenicity must occur in order for a bacterial pathogen to cause disease. Such stages include bacterial adherence to host tissue, bacterial entry into host tissues (usually required), multiplication within host tissues, evasion of host defence mechanisms and the causation of damage. In this study, these key strategies in infection and disease were sought to help assess the pathogenic potential of P. shigelloides (Chapter 3). Twelve isolates of P. shigelloides, obtained from clinical cases of gastroenteritis, were used to infect monolayers of human intestinal epithelial cells in vitro. Ultrastructural analysis demonstrated that P. shigelloides was able to adhere to the microvilli at the apical surface of the epithelial cells and also to the plasma membranes of both apical and basal surfaces. Furthermore, it was demonstrated that these isolates were able to enter intestinal epithelial cells. Internalised bacteria often were confined within vacuoles surrounded by single or multiple membranes. Observation of bacteria within membranebound vacuoles suggests that uptake of P. shigelloides into intestinal epithelial cells occurs via a process morphologically comparable to phagocytosis. Bacterial cells also were observed free in the host cell cytoplasm, indicating that P. shige/loides is able to escape from the surrounding vacuolar membrane and exist within the cytosol of the host. Plesiomonas shigelloides has not only been implicated in gastrointestinal infections, but also in a range of non-intestinal infections such as cholecystitis, proctitis, septicaemia and meningitis. The mechanisms by which P. shigelloides causes these infections are not understood. Previous research was unable to ascertain the pathogenic potential of P. shigel/oides using cells of non-intestinal origin (HEp-2 cells derived from a human larynx carcinoma and Hela cells derived from a cervical carcinoma). However, with the recent findings (from this study) that P. shigelloides can adhere to and enter intestinal cells, it was hypothesised, that P. shigel/oides would be able to enter Hela and HEp-2 cells. Six clinical isolates of P. shigelloides, which previously have been shown to be invasive to intestinally derived Caco-2 cells (Chapter 3) were used to study interactions with Hela and HEp-2 cells (Chapter 4). These isolates were shown to adhere to and enter both nonintestinal host cell lines. Plesiomonas shigelloides were observed within vacuoles surrounded by single and multiple membranes, as well as free in the host cell cytosol, similar to infection by P. shigelloides of Caco-2 cells. Comparisons of the number of bacteria adhered to and present intracellularly within Hela, HEp-2 and Caco-2 cells revealed a preference of P. shigelloides for Caco-2 cells. This study conclusively showed for the first time that P. shigelloides is able to enter HEp-2 and Hela cells, demonstrating the potential ability to cause an infection and/or disease of extra-intestinal sites in humans. Further high resolution ultrastructural analysis of the mechanisms involved in P. shigelloides adherence to intestinal epithelial cells (Chapter 5) revealed numerous prominent surface features which appeared to be involved in the binding of P. shige/loides to host cells. These surface structures varied in morphology from small bumps across the bacterial cell surface to much longer filaments. Evidence that flagella might play a role in bacterial adherence also was found. The hypothesis that filamentous appendages are morphologically expressed when in contact with host cells also was tested. Observations of bacteria free in the host cell cytosol suggests that P. shigelloides is able to lyse free from the initial vacuolar compartment. The vacuoles containing P. shigel/oides within host cells have not been characterised and the point at which P. shigelloides escapes from the surrounding vacuolar compartment has not been determined. A cytochemical detection assay for acid phosphatase, an enzymatic marker for lysosomes, was used to analyse the co-localisation of bacteria-containing vacuoles and acid phosphatase activity (Chapter 6). Acid phosphatase activity was not detected in these bacteria-containing vacuoles. However, the surface of many intracellular and extracellular bacteria demonstrated high levels of acid phosphatase activity, leading to the proposal of a new virulence factor for P. shigelloides. For many pathogens, the efficiency with which they adhere to and enter host cells is dependant upon the bacterial phase of growth. Such dependency reflects the timing of expression of particular virulence factors important for bacterial pathogenesis. In previous studies (Chapter 3 to Chapter 6), an overnight culture of P. shigelloides was used to investigate a number of interactions, however, it was unknown whether this allowed expression of bacterial factors to permit efficient P. shigelloides attachment and entry into human cells. In this study (Chapter 7), a number of clinical and environmental P. shigelloides isolates were investigated to determine whether adherence and entry into host cells in vitro was more efficient during exponential-phase or stationary-phase bacterial growth. An increase in the number of adherent and intracellular bacteria was demonstrated when bacteria were inoculated into host cell cultures in exponential phase cultures. This was demonstrated clearly for 3 out of 4 isolates examined. In addition, an increase in the morphological expression of filamentous appendages, a suggested virulence factor for P. shigel/oides, was observed for bacteria in exponential growth phase. These observations suggest that virulence determinants for P. shigel/oides may be more efficiently expressed when bacteria are in exponential growth phase. This study demonstrated also, for the first time, that environmental water isolates of P. shigelloides were able to adhere to and enter human intestinal cells in vitro. These isolates were seen to enter Caco-2 host cells through a process comparable to the clinical isolates examined. These findings support the hypothesis of a water transmission route for P. shigelloides infections. The results presented in this thesis contribute significantly to our understanding of the pathogenic mechanisms involved in P. shigelloides infections and disease. Several of the factors involved in P. shigelloides pathogenesis have homologues in other pathogens of the human intestine, namely Vibrio, Aeromonas, Salmonella, Shigella species and diarrhoeaassociated strains of Escherichia coli. This study emphasises the relevance of research into Plesiomonas as a means of furthering our understanding of bacterial virulence in general. As well it provides tantalising clues on normal and pathogenic host cell mechanisms.

Discovering, applying and integrating self-knowledge : a grounded theory study of learning in life coaching

Professional coaching is a rapidly expanding field with interdisciplinary roots and broad application. However, despite abundant prescriptive literature, research into the process of coaching, and especially life coaching, is minimal. Similarly, although learning is inherently recognised in the process of coaching, and coaching is increasingly being recognised as a means of enhancing teaching and learning, the process of learning in coaching is little understood, and learning theory makes up only a small part of the evidence-based coaching literature. In this grounded theory study of life coaches and their clients, the process of learning in life coaching across a range of coaching models is examined and explained. The findings demonstrate how learning in life coaching emerged as a process of discovering, applying and integrating self-knowledge, which culminated in the development of self. This process occurred through eight key coaching processes shared between coaches and clients and combined a multitude of learning theory.

An architecture for user configurable mobile collaborative geographic applications

Geographic information is increasingly being touted for use in research and industrial projects. While the technology is now available and affordable, there is a lack of easy to use software that takes advantage of geographic information. This is an important problem because users are often researchers or scientists who have insufficient software skills, and by providing applications that are easier to use, time and financial resources can be taken from training and be better applied to the actual research and development work. A solution for this problem must cater for the user and research needs. In particular it must allow for mobile operation for fieldwork, flexibility or customisability of data input, sharing of data with other tools and collaborative capabilities for the usual teamwork environment. This thesis has developed a new architecture and data model to achieve the solution. The result is the Mobile Collaborative Annotation framework providing an implementation of the new architecture and data model. Mobile Collaborative Mapping implements the framework as a Web 2.0 mashup rich internet application and has proven to be an effective solution through its positive application to a case study with fieldwork scientists. This thesis has contributed to research into mobile computing, collaborative computing and geospatial systems by creating a simpler entry point to mobile geospatial applications, enabling simplified collaboration and providing tangible time savings.

The work of nurse practitioners : a national work sampling study

Aim: This paper is a report of a study of variations in the pattern of nurse practitioner work in a range of service fields and geographical locations, across direct patient care, indirect patient care and service-related activities. Background. The nurse practitioner role has been implemented internationally as a service reform model to improve the access and timeliness of health care. There is a substantial body of research into the nurse practitioner role and service outcomes, but scant information on the pattern of nurse practitioner work and how this is influenced by different service models. --------- Methods: We used work sampling methods. Data were collected between July 2008 and January 2009. Observations were recorded from a random sample of 30 nurse practitioners at 10-minute intervals in 2-hour blocks randomly generated to cover two weeks of work time from a sampling frame of six weeks. --------- Results: A total of 12,189 individual observations were conducted with nurse practitioners across Australia. Thirty individual activities were identified as describing nurse practitioner work, and these were distributed across three categories. Direct care accounted for 36.1% of how nurse practitioners spend their time, indirect care accounted for 32.2% and service-related activities made up 31.9%. --------- Conclusion. These findings provide useful baseline data for evaluation of nurse practitioner positions and the service effect of these positions. However, the study also raises questions about the best use of nurse practitioner time and the influences of barriers to and facilitators of this model of service innovation.

Application of K-tree to document clustering

Digital collections are growing exponentially in size as the information age takes a firm grip on all aspects of society. As a result Information Retrieval (IR) has become an increasingly important area of research. It promises to provide new and more effective ways for users to find information relevant to their search intentions. Document clustering is one of the many tools in the IR toolbox and is far from being perfected. It groups documents that share common features. This grouping allows a user to quickly identify relevant information. If these groups are misleading then valuable information can accidentally be ignored. There- fore, the study and analysis of the quality of document clustering is important. With more and more digital information available, the performance of these algorithms is also of interest. An algorithm with a time complexity of O(n2) can quickly become impractical when clustering a corpus containing millions of documents. Therefore, the investigation of algorithms and data structures to perform clustering in an efficient manner is vital to its success as an IR tool. Document classification is another tool frequently used in the IR field. It predicts categories of new documents based on an existing database of (doc- ument, category) pairs. Support Vector Machines (SVM) have been found to be effective when classifying text documents. As the algorithms for classifica- tion are both efficient and of high quality, the largest gains can be made from improvements to representation. Document representations are vital for both clustering and classification. Representations exploit the content and structure of documents. Dimensionality reduction can improve the effectiveness of existing representations in terms of quality and run-time performance. Research into these areas is another way to improve the efficiency and quality of clustering and classification results. Evaluating document clustering is a difficult task. Intrinsic measures of quality such as distortion only indicate how well an algorithm minimised a sim- ilarity function in a particular vector space. Intrinsic comparisons are inherently limited by the given representation and are not comparable between different representations. Extrinsic measures of quality compare a clustering solution to a “ground truth” solution. This allows comparison between different approaches. As the “ground truth” is created by humans it can suffer from the fact that not every human interprets a topic in the same manner. Whether a document belongs to a particular topic or not can be subjective.

Revisiting 'how we learn' in academia : practice-based learning exchanges in three Australian universities

Ideas of 'how we learn' in formal academic settings have changed markedly in recent decades. The primary position that universities once held on shaping what constitutes learning has come into question from a range of experience-led and situated learning models. Drawing on findings from a study conducted across three Australian universities, the article focuses on the multifarious learning experiences indicative of practice-based learning exchanges such as student placements. Building on both experiential and situated learning theories, the authors found that students can experience transformative and emotional elucidations of learning, that can challenge tacit assumptions and transform the ways they understand the world. It was found that all participants (hosts, students, academics) both teach and learn in these educative scenarios and that, contrary to common (mis)perceptions that academics live in 'ivory towers', they play a crucial role in contributing to learning that takes place in the so-called 'real world'.

The validity of simulators in studying driving behaviours

Driver simulators provide safe conditions to assess driver behaviour and provide controlled and repeatable environments for study. They are a promising research tool in terms of both providing safety and experimentally well controlled environments. There are wide ranges of driver simulators, from laptops to advanced technologies which are controlled by several computers in a real car mounted on platforms with six degrees of freedom of movement. The applicability of simulator-based research in a particular study needs to be considered before starting the study, to determine whether the use of a simulator is actually appropriate for the research. Given the wide range of driver simulators and their uses, it is important to know beforehand how closely the results from a driver simulator match results found in the real word. Comparison between drivers’ performance under real road conditions and in particular simulators is a fundamental part of validation. The important question is whether the results obtained in a simulator mirror real world results. In this paper, the results of the most recently conducted research into validity of simulators is presented.

Applying the open innovation system concept to infrastructure projects

Purpose: The goal of this conceptual paper is to provide tools to help maximise the value delivered by infrastructure projects, by developing methods to increase adoption of innovative products during construction. Methods: The role of knowledge flows in determining innovation adoption rates is conceptually examined. A promising new approach is developed. Open innovation system theory is extended, by reviewing the role of three frameworks: (1) knowledge intermediaries, (2) absorptive capacity and (3) governance arrangements. Originality: We develop a novel open innovation system model to guide further research in the area of adoption of innovation on infrastructure projects. The open innovation system model currently lacks definition of core concepts, especially with regard to the impact of different degrees and types of openness. The three frameworks address this issue and add substance to the open innovation system model, addressing widespread criticism that it is underdeveloped. The novelty of our model is in the combination of the three frameworks to explore the system. These frameworks promise new insights into system dynamics and facilitate the development of new methods to optimise the diffusion of innovation. Practical Implications: The framework will help to reveal gaps in knowledge flows that impede the uptake of innovations. In the past, identifying these gaps has been difficult given the lack of nuance in existing theory. The knowledge maps proposed will enable informed policy advice to effectively harness the power of knowledge networks, increase innovation diffusion and improve the performance of infrastructure projects. The models developed in this paper will be used in planned empirical research into innovation on large scale infrastructure projects in the Australian built environment.

A framework for successful enterprise systems implementation : preliminary findings from a case study

While critical success factors (CSFs) of enterprise system (ES) implementation are mature concepts and have received considerable attention for over a decade, researchers have very often focused on only a specific aspect of the implementation process or a specific CSF. Resultantly, there is (1) little research documented that encompasses all significant CSF considerations and (2) little empirical research into the important factors of successful ES implementation. This paper is part of a larger research effort that aims to contribute to understanding the phenomenon of ES CSFs, and reports on preliminary findings from a case study conducted at a Queensland University of Technology (QUT) in Australia. This paper reports on an empirically derived CSFs framework using a directed content analysis of 79 studies; from top IS outlets, employing the characteristics of the analytic theory, and from six different projects implemented at QUT.

Optimism about safety and group-serving interpretations of safety among pedestrians and cyclists in relation to road use in general and under low light conditions

Drivers are known to be optimistic about their risk of crash involvement, believing that they are less likely to be involved in a crash than other drivers. However, little comparative research has been conducted among other road users. In addition, optimism about crash risk is conceptualised as applying only to an individual’s assessment of his or her personal risk of crash involvement. The possibility that the self-serving nature of optimism about safety might be generalised to the group-level as a cyclist or a pedestrian, i.e., becoming group-serving rather than self-serving, has been overlooked in relation to road safety. This study analysed a subset of data collected as part of a larger research project on the visibility of pedestrians, cyclists and road workers, focusing on a set of questionnaire items administered to 406 pedestrians, 838 cyclists and 622 drivers. The items related to safety in various scenarios involving drivers, pedestrians and cyclists, allowing predictions to be derived about group differences in agreement with items based on the assumption that the results would exhibit group-serving bias. Analysis of the responses indicated that specific hypotheses about group-serving interpretations of safety and responsibility were supported in 22 of the 26 comparisons. When the nine comparisons relevant to low lighting conditions were considered separately, seven were found to be supported. The findings of the research have implications for public education and for the likely acceptance of messages which are inconsistent with current assumptions and expectations of pedestrians and cyclists. They also suggest that research into group-serving interpretations of safety, even for temporary roles rather than enduring groups, could be fruitful. Further, there is an implication that gains in safety can be made by better educating road users about the limitations of their visibility and the ramifications of this for their own road safety, particularly in low light.

Best practice for prevention and treatment of cardiovascular disease through an equity lens : a review

Background: Despite declining rates of cardiovascular disease (CVD) mortality in developed countries, lower socioeconomic groups continue to experience a greater burden of the disease. There are now many evidence-based treatments and prevention strategies for the management of CVD and it is essential that their impact on the more disadvantaged group is understood if socioeconomic inequalities in CVD are to be reduced. Aims: To determine whether key interventions for CVD prevention and treatment are effective among lower socioeconomic groups, to describe barriers to their effectiveness and the potential or actual impact of these interventions on the socioeconomic gradient in CVD. Methods: Interventions were selected from four stages of the CVD continuum. These included smoking reduction strategies, absolute risk assessment, cardiac rehabilitation, secondary prevention medications, and heart failure self-management programmes. Electronic searches were conducted using terms for each intervention combined with terms for socioeconomic status (SES). Results: Only limited evidence was found for the effectiveness of the selected interventions among lower SES groups and there was little exploration of socioeconomic-related barriers to their uptake. Some broad themes and key messages were identified. In the majority of findings examined, it was clear that the underlying material, social and environmental factors associated with disadvantage are a significant barrier to the effectiveness of interventions. Conclusion: Opportunities to reduce socioeconomic inequalities occur at all stages of the CVD continuum. Despite this, current treatment and prevention strategies may be contributing to the widening socioeconomic-CVD gradient. Further research into the impact of best-practice interventions for CVD upon lower SES groups is required.

Interventions for preventing and managing aggressive patients admitted to an acute hospital setting: A systematic review

Background: Violence in health care has been widely reported and health care workers, particularly nurses in acute care settings, are ill-equipped to manage patients who exhibit aggressive traits. Aim: The aim of this systematic review was to establish best practice in the prevention and management of aggressive behaviours in patients admitted to acute hospital settings. Data Sources: An extensive search of the major databases was conducted from 1990 to 2007. The search included published and unpublished studies and papers in English. Review Methods: This review considered any quantitative research study design that evaluated the effectiveness of interventions in the prevention and management of patients who exhibit aggressive behaviours in an acute hospital setting. Each included study was quality assessed by two independent reviewers and data were extracted using the relevant tools developed by the Joanna Briggs Institute. Results: Ten studies met the inclusion criteria and were included in the review. The evidence identified from the studies includes: the benefit of education and training of acute care nurses in aggression management techniques; use of “as required” medications is effective in minimising harm to patients and staff; and that specific interventions such as physical restraint may play a role in managing aggressive behaviours from patients in the acute care setting. Conclusions: This review makes several recommendations for the prevention and management of aggressive behaviours in acute hospital patients. However, due to the lack of high-quality studies conducted in the acute care setting there is huge scope for future research in this area.

Anna Clark (2008) History’s Children: History Wars in the Classroom. UNSW Press, Sydney, 178 pages

“History’s Children” stems from Anna Clark’s 2004 postdoctoral research into the ways in which Australian students connect with the past, and aims at bringing some classroom perspectives into the public debates about Australian history education. Although the title makes reference to the “History Wars”, there is little evidence of contestation, engagement, passion or intellectual excitement in Clark’s conclusions about what happens in history classrooms. Rather, Clark’s small focus groups with 182 high school students in 34 high schools around Australia indicate that “it got a bit dismal hearing student after student being so dismissive of Australian history” (p. 143). Apart from some enthusiasm for the study of Australians at war, a sort of resigned boredom seems to characterise what students have to say about learning Australian history, despite their acknowledgement that it is important to “know about” it.

Towards a comprehensive model of driver aggression : a review of the literature and directions for the future

Driver aggression is a road safety issue of growing concern throughout most highly motorised countries, yet to date there is no comprehensive model that deals with this issue in the road safety area. This paper sets out to examine the current state of research and theory on aggressive driving with a view to incorporating useful developments in the area of human aggression from mainstream psychological research. As a first step, evidence regarding the prevalence and incidence of driver aggression, including the impact of the phenomenon on crash rates is reviewed. Inconsistencies in the definition and operationalisation of driver aggression that have hampered research in the area are noted. Existing models of driver aggression are then identified and the need to distinguish and address the role of intentionality as well as the purpose of perpetrating behaviours within both these and research efforts is highlighted. Drawing on recent findings from psychological research into general aggression, it is argued that progress in understanding driver aggression requires models that acknowledge not only the person-related and situational factors, but the cognitive and emotional appraisal processes involved in driver aggression. An effective model is expected to allow the explanation of not only the likelihood and severity of driver aggression behaviours, but also the escalation of incidents within the context of the road environment.

Detection of anomalies from user profiles generated from system logs

We describe research into the identification of anomalous events and event patterns as manifested in computer system logs. Prototype software has been developed with a capability that identifies anomalous events based on usage patterns or user profiles, and alerts administrators when such events are identified. To reduce the number of false positive alerts we have investigated the use of different user profile training techniques and introduce the use of abstractions to group together applications which are related. Our results suggest that the number of false alerts that are generated is significantly reduced when a growing time window is used for user profile training and when abstraction into groups of applications is used.