The expression, regulation and function of kallikrein 4 in prostate cancer

Prostate cancer is a significant health problem faced by aging men. Currently, diagnostic strategies for the detection of prostate cancer are either unreliable, yielding high numbers of false positive results, or too invasive to be used widely as screening tests. Furthermore, the current therapeutic strategies for the treatment of the disease carry considerable side effects. Although organ confined prostate cancer can be curable, most detectable clinical symptoms occur in advanced disease when primary tumour cells have metastasised to distant sites - usually lymph nodes and bone. Many growth factors and steroids assist the continued growth and maintenance of prostatic tumour cells. Of these mitogens, androgens are important in the development of the normal prostate but are also required to sustain the growth of prostate cancer cells in the early stage of the disease. Not only are androgens required in the early stage of disease, but also many other growth factors and hormones interact to cause uncontrolled proliferation of malignant cells. The early, androgen sensitive phase of disease is followed by an androgen insensitive phase, whereby androgens are no longer required to stimulate the growth of the tumour cells. Growth factors such as transforming growth factor  and  (TGF/), epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), insulin-like growth factors (IGFs), Vitamin D and thyroid hormone have been suggested to be important at this stage of disease. Interestingly, some of the kallikrein family of genes, including prostate specific antigen (PSA), the current serum diagnostic marker for prostate cancer, are regulated by androgens and many of the aforementioned growth factors. The kallikrein gene family is a group of serine proteases that are involved in a diverse range of physiological processes: regulation of local blood flow, angiogenesis, tissue invasion and mitogenesis. The earliest members of the kallikrein gene family (KLK1-KLK3) have been strongly associated with general disease states, such as hypertension, inflammation, pancreatitis and renal disease, but are also linked to various cancers. Recently, this family was extended to include 15 genes (KLK1-15). Several newer members of the kallikrein family have been implicated in the carcinogenesis and tumour metastasis of hormone-dependent cancers such as prostate, breast, endometrial and ovarian cancer. The aims of this project were to investigate the expression of the newly identified kallikrein, KLK4, in benign and malignant prostate tissues, and prostate cancer cell lines. This thesis has demonstrated the elevated expression of KLK4 mRNA transcripts in malignant prostate tissue compared to benign prostates. Additionally, expression of the full length KLK4 transcript was detected in the androgen dependent prostate cancer cell line, LNCaP. Based on the above finding, the LNCaP cell line was chosen to assess the potential regulation of full length KLK4 by androgen, thyroid hormone and epidermal growth factor. KLK4 mRNA and protein was found to be up-regulated by androgen and a combination of androgen and thyroid hormone. Thyroid hormone alone produced no significant change in KLK4 mRNA or protein over the control. Epidermal growth factor treatment also resulted in elevated expression levels of KLK4 mRNA and protein. To assess the potential functional role(s) of KLK4/hK4 in processes associated with tumour progression, full length KLK4 was transfected into PC-3 cells - a prostate cancer cell line originally derived from a secondary bone lesion. The KLK4/hK4 over-expressing cells were assessed for their proliferation, migration, invasion and attachment properties. The KLK4 over-expressing clones exhibited a marked change in morphology, indicative of a more aggressive phenotype. The KLK4 clones were irregularly shaped with compromised adhesion to the growth surface. In contrast, the control cell lines (parent PC-3 and empty vector clones) retained a rounded morphology with obvious cell to cell adhesion, as well as significant adhesion to their growth surface. The KLK4 clones exhibited significantly greater attachment to Collagen I and IV than native PC-3s and empty vector controls. Over a 12 hour period, in comparison to the control cells, the KLK4 clones displayed an increase in migration towards PC-3 native conditioned media, a 3 fold increase towards conditioned media from an osteoblastic cell line (Saos-2) and no change in migration towards conditioned media from neonatal foreskin fibroblast cells or 20% foetal bovine serum. Furthermore, the increase in migration exhibited by the KLK4 clones was partially blocked by the serine protease inhibitor, aprotinin. The data presented in this thesis suggests that KLK4/hK4 is important in prostate carcinogenesis due to its over-expression in malignant prostate tissues, its regulation by hormones and growth factors associated with prostate disease and the functional consequences of over-expression of KLK4/hK4 in the PC-3 cell line. These results indicate that KLK4/hK4 may play an important role in tumour invasion and bone metastasis via increased attachment to the bone matrix protein, Collagen I, and enhanced migration due to soluble factors produced by osteoblast cells. This suggestion is further supported by the morphological changes displayed by the KLK4 over-expressing cells. Overall, this data suggests that KLK4/hK4 should be further studied to more fully investigate the potential value of KLK4/hK4 as a diagnostic/prognostic biomarker or in therapeutic applications.

‘Show Me Your Wiki and I’ll Show you Mine’: Using Online Interactive Media to Improve Academic Writing and Research in a Public Health Under-Graduate Cohort

The number of Internet users in Australia has been steadily increasing, with over 10.9 million people currently subscribed to an internet provider (ABS, 2011). Over the past year, the most avid users of the Internet were 15 – 24 year olds, with approximately 95% accessing the internet on a regular basis (ABS, Social Trends, 2011). While the internet has been described as fundamental to higher education students, social and leisure internet tools are also increasingly being used by these students to generate and maintain their social and professional networks and interactions (Duffy & Bruns 2006). Rapid technological advancements have enabled greater and faster access to information for learning and education (Hemmi et al, 2009; Glassman and Kang, 2011). As such, we sought to integrate interactive, online social media into the assessment profile of a Public Health undergraduate cohort at the Queensland University of Technology (QUT). The aim of this exercise was to engage students to both develop and showcase their research on a range of complex, contemporary health issues within the online forum of Wikispaces (http://www.wikispaces.com/) for review and critique by their peers. We applied Bandura’s Social Learning Theory (SLT) to analyse the interactive processes from which students developed deeper and more sustained learning, and via which their overall academic writing standards were raised. This paper outlines the assessment task, and the students’ feedback on their learning outcomes in relation to the Attentional, Retentional, Motor Reproduction, and Motivational Processes outlined by Bandura in SLT. We conceptualise the findings in a theoretical model, and discuss the implications for this approach within the broader tertiary environment.

Crime is in the air : air pollution and regulation in the UK

This latest briefing by Professor Reece Walters in the What is crime? series, draws attention to an area of harm that is often absent from criminological debate. He highlights the human costs of air pollution and failed attempts to adequately regulate and control such harm. Arguing for a cross disciplinary ‘eco-crime’ narrative, the author calls for greater understanding of the far-reaching consequences of air pollution which could set in train changes which may lead to a ‘more robust and meaningful system of justice’. Describing current arrangements in place to control and regulate air pollution, Walters draws attention to the lack of neutrality in current arrangements and the bias ‘towards the economic imperatives of free trade over and above the centrality of environmental protection’. While attention is often given to direct and individualised instances of ‘crime’, the serious consequences of air pollution are frequently neglected. The negative effects of pollution on health and well-being are often borne by people already experiencing a range of other disadvantages. In a global and national context, it is often the poor who are affected most. Ultimately, political and economic imperatives have historically helped to shape legal and regulatory regimes. Whether this is an inherent flaw in current systems or something that can be overcome in favour of dealing with more wide-ranging harms is an area that requires further discussion and debate.

Incentivising private health insurance through the income tax regime : Capitalising on behavioural models

The Australian income tax regime is generally regarded as a mechanism by which the Federal Government raises revenue, with much of the revenue raised used to support public spending programs. A prime example of this type of spending program is health care. However, a government may also decide that the private sector should provide a greater share of the nation's health care. To achieve such a policy it can bring about change through positive regulation, or it can use the taxation regime, via tax expenditures, not to raise revenue but to steer or influence individuals in its desired direction. When used for this purpose, tax expenditures steer taxpayers towards or away from certain behaviour by either imposing costs on, or providing benefits to them. Within the context of the health sector, the Australian Federal Government deploys social steering via the tax system, with the Medicare Levy Surcharge and the 30 percent Private Health Insurance Rebate intended to steer taxpayer behaviour towards the Government’s policy goal of increasing the amount of health provision through the private sector. These steering mechanisms are complemented by the ‘Lifetime Health Cover Initiative’. This article, through the lens of behavioural economics, considers the ways in which these assorted mechanisms might have been expected to operate and whether they encourage individuals to purchase private health insurance.

If it’s sick, how can we well? : Four criteria for evaluating the health of journalism

This paper seeks to address the highly pervasive discourse that journalism is ‘in crisis’ by outlining four criteria by which we might evaluate the ‘health’ of the practice (measures of both quantity and quality of output). It offers an extremely brief meta-level analysis of existing research, and posits that when judged according to these four criteria, journalism might actually in reasonable health,and that we ought to be far more optimistic about its future. This assessment therefore challenges the ‘business-centric’ evaluation which often dominates discussions (in the media as well as academia) about the profession’s supposedly dire future.

Health Workforce Governance: Improved Access, Good Regulatory Practice, Safer Patients

With increasing recognition of the international market in health professionals and the impact of globalism on regulation, the governance of the health workforce is moving towards greater public engagement and increased transparency. This book discusses the challenges posed by these processes, such as improved access to health services and how structures can be reformed so that good practice is upheld and quality of service and patient safety are ensured.

Evaluation of the skills for preventing injury in youth program : impact on health risk behaviors, injury and school connectedness

Injury is the leading cause of death among young people, and involvement in health risk behaviors, such as alcohol use and transport-related risks, is related to increased risk for injury. Effective health promotion programs for adolescents focus on multiple levels, including relationships with peers and parents, student knowledge, behavior and attitudes, and school-level factors such as school connectedness. This study describes the pilot evaluation of a comprehensive, multi-level injury prevention program for 13-14 year old adolescents, targeting change in injury associated with transport and alcohol risks. The program, called Skills for Preventing Injury in Youth (SPIY), incorporates two primary elements: an 8-week, teacher delivered attitude and behavior change curriculum with peer protection and first aid messages; and professional development for program teachers focusing on strategies to increase students’ connectedness to school. Five Australian high schools were recruited for the pilot evaluation research, with three being assigned to receive intervention components and two assigned as curriculum-as-usual controls. In the intervention schools, 118 Year 8 students participated in surveys at baseline, with 105 completing surveys at follow up, six months following the intervention. In the control schools, 196 Year 8 students completed surveys at baseline and 207 at follow up. Survey measures included self-reported injury, risk taking behavior and school connectedness. Results showed that students in the control schools were significantly more likely to report riding bikes without helmets, riding with dangerous drivers, having driven cars on the road, and using alcohol six months after the program, while the intervention group showed no such increase in these behaviors. Additionally, students in the control schools were significantly more likely to report having had pedestrian-related injuries at follow up than they were at the baseline measurement, while intervention school students showed no change. There was also a trend observed in terms of a decrease in bicycle related injuries among intervention school students, compared with a slight increasing trend in bicycle injuries among control students. Overall, scores on the school connectedness scale decreased significantly from baseline to follow up for both intervention and control students, however measurement limitations may have impacted on results relating to students’ connectedness. Overall, the SPIY program has shown promising results in regards to prevention of students’ health risk behavior and injuries. Evidence suggests that the curriculum component was important; however there was limited evidence to suggest that teacher training in school connectedness strategies contributed to these promising results. While school connectedness may be an important factor to target in risk and injury prevention programs, programs may need to incorporate whole-of-school strategies or target a broader range of teachers than were selected for the current research.

Regulation of proteasome structure and function

Proteasomes are cylindrical particles made up of a stack of four heptameric rings. In animal cells the outer rings are made up of 7 different types of alpha subunits and the inner rings are composed of 7 out of 10 possible different beta subunits. Regulatory complexes can bind to the ends of the cylinder.We have investigated aspects of the assembly, activity and subunit composition of core proteasome particles and 26S proteasomes, the localization of proteasome subpopulations, and the possible role of phosphorylation in determining proteasome localization, activities and association with regulatory components.

Catalytic components of proteasomes and the regulation of proteinase activity

The proteasome (multicatalytic proteinase complex) is a large multimeric complex which is found in the nucleus and cytoplasm of eukaryotic cells. It plays a major role in both ubiquitin-dependent and ubiquitin-independent nonlysosomal pathways of protein degradation. Proteasome subunits are encoded by members of the same gene family and can be divided into two groups based on their similarity to the c~ and /3 subunits of the simpler proteasome isolated from Thermoplasma acidophilum. Proteasomes have a cylindrical structure composed of four rings of seven subunits. The 26S form of the proteasome, which is responsible for ubiquitin-dependent proteolysis, contains additional regulatory complexes. Eukaryotic proteasomes have multiple catalytic activities which are catalysed at distinct sites. Since proteasomes are unrelated to other known proteases, there are no clues as to which are the catalytic components from sequence alignments. It has been assumed from studies with yeast mutants that /3-type subunits play a catalytic role. Using a radiolabelled peptidyl chloromethane inhibitor of rat liver proteasomes we have directly identified RC7 as a catalytic component. Interestingly, mutants in Prel, the yeast homologue of RC7, have already been reported to have defective chymotrypsin-like activity. These results taken together confirm a direct catalytic role for these/3-type subunits. Proteasome activities are sensitive to conformational changes and there are several ways in which proteasome function may be modulated in vivo. Our recent studies have shown that in animal cells at least two proteasome subunits can undergo phosphorylation, the level of which is likely to be important for determining proteasome localization, activity or ability to form larger complexes. In addition, we have isolated two isoforms of the 26S proteinase.

Using reflective practice assessment in the first year of law to encourage a positive professional identity and promote law student well-being

In this paper we argue that intentional curriculum design in the first year of law should encourage law students to develop an emergent sense of a positive professional identity. When first year law students engage with a nascent notion of a positive professional identity, their well-being is supported because their studies are informed and contextualised by a sense of purpose for their future professional life. In a first year law subject run for the first time at the QUT Law School in 2011, reflective practice was successfully used to achieve these goals. The paper discusses the subject, the opportunity of using reflective practice to teach a positive sense of professional identity, and some student perspectives on the subject’s design.

An action research approach for the professional development of Vietnamese nurse educators

Nurse education in Viet Nam is undergoing substantial reform. In order to facilitate the change, in 2007 the Viet Nam Nurses Association formed a collaborative partnership with the School of Nursing and Midwifery at an Australia university. This collaboration gave rise to the Viet Nam Nursing Capacity Building Project under the leadership of Professor Genevieve Gray, funded by the Atlantic Philanthropies. The new four year competency based nursing curriculum frame is expected to be implemented in September 2011 following approval by the Viet Nam Ministry of Education. The focus of this paper is the Teaching Fellowships Program, an initiative of the Viet Nam Nursing Capacity Building Project, developed to help meet the challenges associated with leading and dealing with the curriculum change. The paper explores the development of the program and justifies an action research approach, illuminates key issues, and briefly refers to changes to the next fellowship program.

Molecular characterisation of environmental enterococci derived from water samples and assessment of associated public health hazards

Enterococci are versatile Gram-positive bacteria that can survive under extreme conditions. Most enterococci are non-virulent and found in the gastrointestinal tract of humans and animals. Other strains are opportunistic pathogens that contribute to a large number of nosocomial infections globally. Epidemiological studies demonstrated a direct relationship between the density of enterococci in surface waters and the risk of swimmer-associated gastroenteritis. The distribution of infectious enterococcal strains from the hospital environment or other sources to environmental water bodies through sewage discharge or other means, could increase the prevalence of these strains in the human population. Environmental water quality studies may benefit from focusing on a subset of Enterococcus spp. that are consistently associated with sources of faecal pollution such as domestic sewage, rather than testing for the entire genus. E. faecalis and E. faecium are potentially good focal species for such studies, as they have been consistently identified as the dominant Enterococcus spp. in human faeces and sewage. On the other hand enterococcal infections are predominantly caused by E. faecalis and E. faecium. The characterisation of E. faecalis and E. faecium is important in studying their population structures, particularly in environmental samples. In developing and implementing rapid, robust molecular genotyping techniques, it is possible to more accurately establish the relationship between human and environmental enterococci. Of particular importance, is to determine the distribution of high risk enterococcal clonal complexes, such as E. faecium clonal complex 17 and E. faecalis clonal complexes 2 and 9 in recreational waters. These clonal complexes are recognized as particularly pathogenic enterococcal genotypes that cause severe disease in humans globally. The Pimpama-Coomera watershed is located in South East Queensland, Australia and was investigated in this study mainly because it is used intensively for agriculture and recreational purposes and has a strong anthropogenic impact. The primary aim of this study was to develop novel, universally applicable, robust, rapid and cost effective genotyping methods which are likely to yield more definitive results for the routine monitoring of E. faecalis and E. faecium, particularly in environmental water sources. To fullfill this aim, new genotyping methods were developed based on the interrogation of highly informative single nucleotide polymorphisms (SNPs) located in housekeeping genes of both E. faecalis and E. faecium. SNP genotyping was successfully applied in field investigations of the Coomera watershed, South-East Queensland, Australia. E. faecalis and E. faecium isolates were grouped into 29 and 23 SNP profiles respectively. This study showed the high longitudinal diversity of E. faecalis and E. faecium over a period of two years, and both human-related and human-specific SNP profiles were identified. Furthermore, 4.25% of E. faecium strains isolated from water was found to correspond to the important clonal complex-17 (CC17). Strains that belong to CC17 cause the majority of hospital outbreaks and clinical infections globally. Of the six sampling sites of the Coomera River, Paradise Point had the highest number of human-related and human-specific E. faecalis and E. faecium SNP profiles. The secondary aim of this study was to determine the antibiotic-resistance profiles and virulence traits associated with environmental E. faecalis and E. faecium isolates compared to human pathogenic E. faecalis and E. faecium isolates. This was performed to predict the potential health risks associated with coming into contact with these strains in the Coomera watershed. In general, clinical isolates were found to be more resistant to all the antibiotics tested compared to water isolates and they harbored more virulence traits. Multi-drug resistance was more prevalent in clinical isolates (71.18% of E. faecalis and 70.3 % of E. faecium) compared to water isolates (only 5.66 % E. faecium). However, tetracycline, gentamicin, ciprofloxacin and ampicillin resistance was observed in water isolates. The virulence gene esp was the most prevalent virulence determinant observed in clinical isolates (67.79% of E. faecalis and 70.37 % of E. faecium), and this gene has been described as a human-specific marker used for microbial source tracking (MST). The presence of esp in water isolates (16.36% of E. faecalis and 19.14% of E. faecium) could be indicative of human faecal contamination in these waterways. Finally, in order to compare overall gene expression between environmental and clinical strains of E. faecalis, a comparative gene hybridization study was performed. The results of this investigation clearly demonstrated the up-regulation of genes associated with pathogenicity in E. faecalis isolated from water. The expression study was performed at physiological temperatures relative to ambient temperatures. The up-regulation of virulence genes demonstrates that environmental strains of E. faecalis can pose an increased health risk which can lead to serious disease, particularly if these strains belong to the virulent CC17 group. The genotyping techniques developed in this study not only provide a rapid, robust and highly discriminatory tool to characterize E. faecalis and E. faecium, but also enables the efficient identification of virulent enterococci that are distributed in environmental water sources.

The assessment of mental health in children with intellectual disability

Aim: Individuals with intellectual disability (ID) have higher rates of mental health problems than the general population. Assessment tends to rely heavily on self-report, but persons with ID often have difficulties in identifying and describing their own thoughts and feelings. Measures that are psychometrically sound with typically developing populations may not be as robust in samples with ID. The aim of the current study was to examine a range of self-report measures for assessing the mental health of children with ID, and to consider the appropriateness of minor modifications to those instruments. Method: The participants were 58 children with ID (mean 11.7 years) attending Year 6 in mainstream primary schools. At the first time point they completed four established measures of depression, anxiety and mood. Minor modifications were made to wording and format at re-administration six months later. Results: Internal consistency varied considerably across measures. Modifications resulted in small or no improvements, but the results were relatively consistent over time and across similar measures. Some gender differences were evident. Conclusions: The findings confirm the difficulties that children with ID may have when responding to self-report measures of mental health, and suggest that care should be taken in choice of instruments. While modifications can produce small improvements, it is clear that more robust measures of mental health are needed for persons with ID.