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.

Cultural diversity, cultural networks and trade : international cultural policy debate

This article sketches some of the ways in which the language and concepts of cultural diversity are being taken up internationally. The debate has been driven in part by concerns about the treatment of cultural goods, services and knowledge in trade agreements. But it also involves larger questions about the role of the state, the role of non-state actors in domestic policy formation, and the shape and function of international policy communities comprising both state and non-state actors. The extent of the discussion of cultural diversity internationally is described through new formal and informal cultural networks and work towards an international instrument for cultural diversity to lay our ground rules for international trade, cultural exchange and policy principles to guide governmental responsibilities. The article concludes with analysis of some of these new networks, and investigates why Canada has been so prominent in these international efforts.

Introduction: Culture, trade, services

Ben Goldsmith and Christina Spurgeon introduce the special 'Culture, trade, services' issue of Media International Australia.

Scandals, public inquiries and health professional regulation

Since 1980 there has been an increasing incidence of the use of public inquiries as a process through which scandals raising patient safety and health care quality concerns can be subject to highly public scrutiny. The use of public inquiries and their impact on the governance of health or social systems, especially around issues of patient or client safety, has been examined by a number of commentators (Butler and Drakeford 2003, Masso and Eager 2009, Stanley and Manthorpe 2004, Walshe and Higgins 2002) but public inquiries into scandals in the health system also raise a question about the impact of these inquiries on public perceptions about the adequacy of the various mechanisms for health professional regulation...

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.

Field-of-view, detection range, and false alarm trade-offs in vision-based aircraft detection

This paper presents a survey of previously presented vision based aircraft detection flight test, and then presents new flight test results examining the impact of camera field-of view choice on the detection range and false alarm rate characteristics of a vision-based aircraft detection technique. Using data collected from approaching aircraft, we examine the impact of camera fieldof-view choice and confirm that, when aiming for similar levels of detection confidence, an improvement in detection range can be obtained by choosing a smaller effective field-of-view (in terms of degrees per pixel).

Environmental reporting in the Australian mining industry : complying with regulation or meeting international best practice?

It is nearly 10 years since the introduction of s 299(1)(f) Corporations Act , which requires the disclosure of information regarding a company's environmental performance within its annual report. This provision has generated considerable debate in the years since its introduction, fundamentally between proponents of either a voluntary or mandatory environmental reporting framework. This study examines the adequacy of the current regulatory framework. The environmental reporting practices of 24 listed companies in the resources industries are assessed relative to a standard set by the Global Reporting Initiative (GRI) Sustainability Reporting Guidelines. These Guidelines are argued to represent "international best practice" in environmental reporting and a "scorecard" approach is used to score the quality of disclosure according to this voluntary benchmark. Larger companies in the sample tend to report environmental information over and above the level required by legislation. Some, but not all companies present a stand-alone environmental/sustainability report. However, smaller companies provide minimal information in compliance with s 299(1)(f) . The findings indicate that "international best practice" environmental reporting is unlikely to be achieved by Australian companies under the current regulatory framework. In the current regulatory environment that scrutinises s 299(1)(f) , this article provides some preliminary evidence of the quality of disclosures generated in the Australian market.

The influence of service quality and trade show effectiveness on post-show purchase intention

Purpose The purpose of this paper is to develop and test an integrative services framework to investigate the role of perceived trade show effectiveness on overall trade show service outcome, conceptualised as the intention to purchase a related product after, rather than during, a show. Design/methodology/approach Drawing on the services marketing and trade show literature, the authors test a model of trade show effectiveness with data collected from 592 attendees at a major automotive trade show in a large metropolitan centre. Findings Results show that improving trade show visitors' perceived service quality positively affects visitor perceptions of trade show effectiveness. Furthermore, both trade show effectiveness and service quality directly influence future purchase intention. Research limitations/implications Employing a services theoretical framework to evaluate trade show visitor experiences provides an alternative to the traditional marketing communications approach. By viewing such visits as service encounters, managers must inevitably consider the effects of service quality and service outcomes in determining the likely success of their shows. The study primarily focuses on one large consumer show and therefore does not constitute a complete, nor necessarily representative, sample of the trade show industry. Originality/value The original contribution of the paper stems from the paucity of research conceptualising trade shows as services and the comparative lack of emphasis placed on visitors rather than exhibitors in the literature. The research not only has utility for trade show organisers but also provides necessary theory-based research in the trade show domain.