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.

Vision-based classification of solder joint defects

Inspection of solder joints has been a critical process in the electronic manufacturing industry to reduce manufacturing cost, improve yield, and ensure product quality and reliability. The solder joint inspection problem is more challenging than many other visual inspections because of the variability in the appearance of solder joints. Although many research works and various techniques have been developed to classify defect in solder joints, these methods have complex systems of illumination for image acquisition and complicated classification algorithms. An important stage of the analysis is to select the right method for the classification. Better inspection technologies are needed to fill the gap between available inspection capabilities and industry systems. This dissertation aims to provide a solution that can overcome some of the limitations of current inspection techniques. This research proposes two inspection steps for automatic solder joint classification system. The “front-end” inspection system includes illumination normalisation, localization and segmentation. The illumination normalisation approach can effectively and efficiently eliminate the effect of uneven illumination while keeping the properties of the processed image. The “back-end” inspection involves the classification of solder joints by using Log Gabor filter and classifier fusion. Five different levels of solder quality with respect to the amount of solder paste have been defined. Log Gabor filter has been demonstrated to achieve high recognition rates and is resistant to misalignment. Further testing demonstrates the advantage of Log Gabor filter over both Discrete Wavelet Transform and Discrete Cosine Transform. Classifier score fusion is analysed for improving recognition rate. Experimental results demonstrate that the proposed system improves performance and robustness in terms of classification rates. This proposed system does not need any special illumination system, and the images are acquired by an ordinary digital camera. In fact, the choice of suitable features allows one to overcome the problem given by the use of non complex illumination systems. The new system proposed in this research can be incorporated in the development of an automated non-contact, non-destructive and low cost solder joint quality inspection system.

Tow Takes on the Critical

This important volume on the critical pedagogical approach addresses such topics as critical multiculturalism, gender and language learning, and popular culture. Critical pedagogies are instructional approaches aimed at transforming existing social relations in the interest of greater equity in schools and communities. This paperback edition on the pedagogical approach addresses such topics as critical multiculturalism, gender and language learning, and popular culture. Committed to language education that contributes to social justice - and the political, economic, and sociocultural changes such justice requires - the contributors explore the meaning of creating equitable and critical instructional practices, by exploring diverse representations of knowledge. In addition, recommendations are made for further research, teacher education, and critical testing. Graduate students and researchers in TESOL, applied linguistics, and education will find this volume a thought-provoking and comprehensive presentation of theory and practice in this important new area of scholarship.

Changes in Level of Virus Accumulation and Incidence of Infection are Critical in the Characterization of Rice Tungro Bacilliform Virus (RTBV) Resistance in Rice

Analysis by enzyme-linked immunosorbent assay showed that Rice tungro bacilliform virus (RTBV) accumulated in a cyclic pattern from early to late stages of infection in tungro-susceptible variety, Taichung Native 1 (TN1), and resistant variety, Balimau Putih, singly infected with RTBV or co-infected with RTBV+Rice tungro spherical virus (RTSV). These changes in virus accumulation resulted in differences in RTBV levels and incidence of infection. The virus levels were expressed relative to those of the susceptible variety and the incidence of infection was assessed at different weeks after inoculation. At a particular time point, RTBV levels in TN1 or Balimau Putih singly infected with RTBV were not significantly different from the virus level in plants co-infected with RTBV+RTSV. The relative RTBV levels in Balimau Putih either singly infected with RTBV or co-infected with RTBV+RTSV were significantly lower than those in TN1. The incidence of RTBV infection varied at different times in Balimau Putih but not in TN1, and to determine the actual infection, the number of plants that became infected at least once anytime during the 4wk observation period was considered. Considering the changes in RTBV accumulation, new parameters for analyzing RTBV resistance were established. Based on these parameters, Balimau Putih was characterized having resistance to virus accumulation although the actual incidence of infection was >75%.

After the marketplace: evidence, social science and educational research

This paper is an essay on the state of Australian education that frames new directions for educational research. It outlines three challenges faced by Australian educators: highly spatialised poverty with particularly strong mediating effects on primary school education; the need for intellectual and critical depth in pedagogy, with a focus in the upper primary and middle years; and the need to reinvent senior schooling to address emergent pathways from school to work and civic life. It offers a narrative description of the dynamics of policy making in Australia and North America and argues for an evidence-based approach to social and educational policy – but one quite unlike current test and market-based approaches. Instead, it argues for a multidisciplinary approach to a broad range of empirical and case-based evidence that subjects these to critical, hermeneutic social sciences. Such an approach would join educational policy with educational research, and broader social, community and governmental action with the aim of reorganising and redistributing material, cultural and social resources.