Crosstalk between developmental and tumour-specific signalling pathways : kallikrein-related serine peptidases and nodal in prostrate cancer

Prostate cancer is an important male health issue. The strategies used to diagnose and treat prostate cancer underscore the cell and molecular interactions that promote disease progression. Prostate cancer is histologically defined by increasingly undifferentiated tumour cells and therapeutically targeted by androgen ablation. Even as the normal glandular architecture of the adult prostate is lost, prostate cancer cells remain dependent on the androgen receptor (AR) for growth and survival. This project focused on androgen-regulated gene expression, altered cellular differentiation, and the nexus between these two concepts. The AR controls prostate development, homeostasis and cancer progression by regulating the expression of downstream genes. Kallikrein-related serine peptidases are prominent transcriptional targets of AR in the adult prostate. Kallikrein 3 (KLK3), which is commonly referred to as prostate-specific antigen, is the current serum biomarker for prostate cancer. Other kallikreins are potential adjunct biomarkers. As secreted proteases, kallikreins act through enzyme cascades that may modulate the prostate cancer microenvironment. Both as a panel of biomarkers and cascade of proteases, the roles of kallikreins are interconnected. Yet the expression and regulation of different kallikreins in prostate cancer has not been compared. In this study, a spectrum of prostate cell lines was used to evaluate the expression profile of all 15 members of the kallikrein family. A cluster of genes was co-ordinately expressed in androgenresponsive cell lines. This group of kallikreins included KLK2, 3, 4 and 15, which are located adjacent to one another at the centromeric end of the kallikrein locus. KLK14 was also of interest, because it was ubiquitously expressed among the prostate cell lines. Immunohistochemistry showed that these 5 kallikreins are co-expressed in benign and malignant prostate tissue. The androgen-regulated expression of KLK2 and KLK3 is well-characterised, but has not been compared with other kallikreins. Therefore, KLK2, 3, 4, 14 and 15 expression were all measured in time course and dose response experiments with androgens, AR-antagonist treatments, hormone deprivation experiments and cells transfected with AR siRNA. Collectively, these experiments demonstrated that prostatic kallikreins are specifically and directly regulated by the AR. The data also revealed that kallikrein genes are differentially regulated by androgens; KLK2 and KLK3 were strongly up-regulated, KLK4 and KLK15 were modestly up-regulated, and KLK14 was repressed. Notably, KLK14 is located at the telomeric end of the kallikrein locus, far away from the centromeric cluster of kallikreins that are stimulated by androgens. These results show that the expression of KLK2, 3, 4, 14 and 15 is maintained in prostate cancer, but that these genes exhibit different responses to androgens. This makes the kallikrein locus an ideal model to investigate AR signalling. The increasingly dedifferentiated phenotype of aggressive prostate cancer cells is accompanied by the re-expression of signalling molecules that are usually expressed during embryogenesis and foetal tissue development. The Wnt pathway is one developmental cascade that is reactivated in prostate cancer. The canonical Wnt cascade regulates the intracellular levels of β-catenin, a potent transcriptional co-activator of T-cell factor (TCF) transcription factors. Notably, β-catenin can also bind to the AR and synergistically stimulate androgen-mediated gene expression. This is at the expense of typical Wnt/TCF target genes, because the AR:β-catenin and TCF:β-catenin interactions are mutually exclusive. The effect of β-catenin on kallikrein expression was examined to further investigate the role of β-catenin in prostate cancer. Stable knockdown of β-catenin in LNCaP prostate cancer cells attenuated the androgen-regulated expression of KLK2, 3, 4 and 15, but not KLK14. To test whether KLK14 is instead a TCF:β-catenin target gene, the endogenous levels of β-catenin were increased by inhibiting its degradation. Although KLK14 expression was up-regulated by these treatments, siRNA knockdown of β-catenin demonstrated that this effect was independent of β-catenin. These results show that β-catenin is required for maximal expression of KLK2, 3, 4 and 15, but not KLK14. Developmental cells and tumour cells express a similar repertoire of signalling molecules, which means that these different cell types are responsive to one another. Previous reports have shown that stem cells and foetal tissues can reprogram aggressive cancer cells to less aggressive phenotypes by restoring the balance to developmental signalling pathways that are highly dysregulated in cancer. To investigate this phenomenon in prostate cancer, DU145 and PC-3 prostate cancer cells were cultured on matrices pre-conditioned with human embryonic stem cells (hESCs). Soft agar assays showed that prostate cancer cells exposed to hESC conditioned matrices had reduced clonogenicity compared with cells harvested from control matrices. A recent study demonstrated that this effect was partially due to hESC-derived Lefty, an antagonist of Nodal. A member of the transforming growth factor β (TGFβ) superfamily, Nodal regulates embryogenesis and is re-expressed in cancer. The role of Nodal in prostate cancer has not previously been reported. Therefore, the expression and function of the Nodal signalling pathway in prostate cancer was investigated. Western blots confirmed that Nodal is expressed in DU145 and PC-3 cells. Immunohistochemistry revealed greater expression of Nodal in malignant versus benign glands. Notably, the Nodal inhibitor, Lefty, was not expressed at the mRNA level in any prostate cell lines tested. The Nodal signalling pathway is functionally active in prostate cancer cells. Recombinant Nodal treatments triggered downstream phosphorylation of Smad2 in DU145 and LNCaP cells, and stably-transfected Nodal increased the clonogencity of LNCaP cells. Nodal was also found to modulate AR signalling. Nodal reduced the activity of an androgen-regulated KLK3 promoter construct in luciferase assays and attenuated the endogenous expression of AR target genes including prostatic kallikreins. These results demonstrate that Nodal is a novel example of a developmental signalling molecule that is reexpressed in prostate cancer and may have a functional role in prostate cancer progression. In summary, this project clarifies the role of androgens and changing cellular differentiation in prostate cancer by characterising the expression and function of the downstream genes encoding kallikrein-related serine proteases and Nodal. Furthermore, this study emphasises the similarities between prostate cancer and early development, and the crosstalk between developmental signalling pathways and the AR axis. The outcomes of this project also affirm the utility of the kallikrein locus as a model system to monitor tumour progression and the phenotype of prostate cancer cells.

The body literate: Discourse and inscription in early literacy

The collection will cover all the major fields of discourse studies: including, grammar, stylistics, conversation analysis, narrative analysis, argumentation, psychology of comprehension, ethnography of speaking, and media. It will include classic articles, work from the top scholars in the field, and reflect all the significant debates.

Foundations of technology development, innovation and competitiveness in the globalised knowledge economy

With the growth of high-technology industries and knowledge intensive services, the pursuit of industrial competitiveness has progressed from a broad concern with the processes of industrialisation to a more focused analysis of the factors explaining cross-national variation in the level of participation in knowledge industries. From an examination of cross-national data, the paper develops the proposition that particular elements of the domestic science, technology and industry infrastructure—such as the stock of knowledge and competence in the economy, the capacity for learning and generation of new ideas and the capacity to commercialise new ideas—vary cross-nationally and are related to the level of participation of a nation in knowledge intensive activities. Existing understandings of the role of the state in promoting industrial competitiveness might be expanded to incorporate an analysis of the contribution of the state through the building of competencies in science, technology and industry. Keywords: Knowledge; economy; comparative public policy; innovation; science and technology policy

Standards, teacher judgement and moderation in contexts of national curriculum and assessment reform

This paper puts forward a proposal for reviewing the role and purpose of standards in the context of national curriculum and assessment reform more generally. It seeks to commence the much-needed conversation about standards in the work of teachers as distinct from large-scale testing companies and the policy personnel responsible for reporting. Four key conditions that relate to the effective use of standards to measure improvement and support learning are analysed: clarity about purpose and function; understanding of the representation of standards; moderation practice; and the assessment community. The Queensland experience of the use of standards, teacher judgement and moderation is offered to identify what is educationally preferable in terms of their use and their relationships to curriculum, improvement and accountability. The article illustrates how these practices have recently been challenged by emerging political constraints related to the Australian Government’s implementation of national testing and national partnership funding arrangements tied to the performance of students at or below minimum standards.

Nitrogen pools and fluxes in grassland soils sequestering carbon

Carbon sequestration in agricultural, forest, and grassland soils has been promoted as a means by which substantial amounts of CO2 may be removed from the atmosphere, but few studies have evaluated the associated impacts on changes in soil N or net global warming potential (GWP). The purpose of this research was to ( 1) review the literature to examine how changes in grassland management that affect soil C also impact soil N, ( 2) assess the impact of different types of grassland management on changes in soil N and rates of change, and (3) evaluate changes in N2O fluxes from differently managed grassland ecosystems to assess net impacts on GWP. Soil C and N stocks either both increased or both decreased for most studies. Soil C and N sequestration were tightly linked, resulting in little change in C: N ratios with changes in management. Within grazing treatments N2O made a minor contribution to GWP (0.1-4%), but increases in N2O fluxes offset significant portions of C sequestration gains due to fertilization (10-125%) and conversion (average = 27%). Results from this work demonstrate that even when improved management practices result in considerable rates of C and N sequestration, changes in N2O fluxes can offset a substantial portion of gains by C sequestration. Even for cases in which C sequestration rates are not entirely offset by increases in N2O fluxes, small increases in N2O fluxes can substantially reduce C sequestration benefits. Conversely, reduction of N2O fluxes in grassland soils brought about by changes in management represents an opportunity to reduce the contribution of grasslands to net greenhouse gas forcing.

Role and purpose of standards in the context of national curriculum and assessment reform for accountability, improvement and equity in student learning

In this article our starting point is the current context of national curriculum change and intense speculation about the assessment, standards and reporting. It is written against a background of accountability measures and improvement imperatives, and focuses attention on standards as offering representations of quality. We understand standards to be constructs that aim to achieve public credibility and utility. Further, they can be examined for the purposes they seek to serve and also their expected functions. Fitness for purpose is therefore a useful notion in considering the nature of standards. Our interest in the discussion is the ‘fit’ between how standards are formulated and how they are used in practice, by whom and for what purposes. A related interest is in the matter of how standards can be harnessed to realise improvement.

The importance of environmental issues and costs in Life Cycle Cost Analysis (LCCA) for highway projects

Sustainability has been increasingly recognised as an integral part of highway infrastructure development. In practice however, the fact that financial return is still a project’s top priority for many, environmental aspects tend to be overlooked or considered as a burden, as they add to project costs. Sustainability and its implications have a far-reaching effect on each project over time. Therefore, with highway infrastructure’s long-term life span and huge capital demand, the consideration of environmental cost/ benefit issues is more crucial in life-cycle cost analysis (LCCA). To date, there is little in existing literature studies on viable estimation methods for environmental costs. This situation presents the potential for focused studies on environmental costs and issues in the context of life-cycle cost analysis. This paper discusses a research project which aims to integrate the environmental cost elements and issues into a conceptual framework for life cycle costing analysis for highway projects. Cost elements and issues concerning the environment were first identified through literature. Through questionnaires, these environmental cost elements will be validated by practitioners before their consolidation into the extension of existing and worked models of life-cycle costing analysis (LCCA). A holistic decision support framework is being developed to assist highway infrastructure stakeholders to evaluate their investment decision. This will generate financial returns while maximising environmental benefits and sustainability outcome.

Computer simulation and modeling in railway applications

An electrified railway system includes complex interconnections and interactions of several subsystems. Computer simulation is the only viable means for system evaluation and analysis. This paper discusses the difficulties and requirements of effective simulation models for this specialized industrial application; and the development of a general-purpose multi-train simulator.

Effect of the Summer Months on Body Weight and Composition in College Women

BACKGROUND AND OBJECTIVES: College students and young adults are experiencing the greatest increases in rates of obesity, and 20% of college students are classified as obese. The objective of this study was to compare changes and rates of change in body weight and body composition between the freshman academic year and the summer after the freshman year among female college students. METHODS: Participants were recruited early in their freshman year of college to participate in a prospective longitudinal study examining changes in body weight and composition over the college years. Height and weight were measured, and body composition was assessed using dual energy x-ray absorptiometry (DEXA) at the beginning and end of the freshman year. Upon return from the summer for their sophomore year, participants returned to have all measurements repeated. Sixty-nine female participants completed all three visits. RESULTS: Body weight increased 1.3 kg during the academic period and an additional 0.1 kg during the summer period. Body mass index (BMI) increased between the first two visits but did not change between the last two visits. However, percent fat increased at each visit. Fat-free mass significantly increased 0.5 kg over the academic year but decreased by 1.1 kg over the summer (p<0.05). Greater rates of change were detected in percent fat, fat-free mass, and BMI during the summer compared with the academic year (p<0.05). CONCLUSIONS: Differences in body composition between the academic and summer periods may reflect changes in living situations between these periods. Unfavorable changes during the summer suggest the need to promote healthy lifestyles to freshman women before they leave campus for the summer