The expression of ADAM-9 and -10 in prostate cancer and their regulation by dihydrotestosterone, insulin-like growth Factor-1 and epidermal growth factor

Prostrate Cancer(PCa)is the most common cause of cancer death amongst Western males. PCa occurs in two distinct stages. In its early stage, growth and development is dependent primarily on male sex hormones (androgens) such as testosterone, although other growth factors have roles maintaining PCa cell survival in this stage. In the later stage of PCa development, growth and.maintenance is independent of androgen stimulation and growth factors including Insulin-like Growth Factor -1 (IGf.:·l) and Epidermal Growth Factor (EGF) are thought to have more crucial roles in cell survival and PCa progression. PCa, in its late stages, is highly aggressive and metastatic, that is, tumorigenic cells migrate from the primary site of the body (prostate) and travel via the systemic and lymphatic circulation, residing and colonising in the bone, lymph node, lung, and in more rare cases, the brain. Metastasis involves both cell migration and tissue degradation activities. The degradation of the extracellular matrix (ECM), the tissue surrounding the organ, is mediated in part by members of a family of 26 proteins called the Matrix Metalloproteases (MMPs), whilst ceil adhesion molecules, of which proteins known as Integrins are included, mediate ce11 migration. A family of proteins known as the ADAMs (A Disintegrin . And Metalloprotease domain) were a recently characterised family at the commencement of this study and now comprise 34 members. Because of their dual nature, possessing an active metaiioprotease domain, homologous to that of the MMPs, and an integrin-binding domain capable of regulating cell-cell and cell-ECM contacts, it was thought likely that members of the ADAMs family may have implications for the progression of aggressive cancers such as those ofthe prostate. This study focussed on two particular ADAMs -9 and -10. ADAM-9 has an active metalloprotease domain, which has been shown to degrade constituents of the ECM, including fibronectin, in vitro. It also has an integrin-binding capacity through association with key integrins involved in PCa progression, such as a6~1. ADAM-10 has no such integrin binding activities, but its bovine orthologue, MADM, is able to degrade coHagen type IV, a major component of basement membranes. It is likely human ADAM-10 has the same activity. It is also known to cleave Ll -a protein involved in cell anchorage activities - and collagen type XVII - which is a principal component of the hemidesmosomes of cellular tight junctions. The cleavage of these proteins enables the cell to be released from the surrounding environment and commence migratory activities, as required in metastasis. Previous studies in this laboratory showed the mRNA expression of the five ADAMs -9,- 10, -11, -15 and -17 in PCa cell lines, characteristic of androgen-dependent and androgen independent disease. These studies were furthered by the characterisation of AD AM-9, -10 and -17 mRNA regulation by Dihydrotestosterone (DHT) in the androgen-responsive cell line (LNCaP). ADAM-9 and -10 mRNA levels were elevated in response to DHT stimulation. Further to these observations, the expression of ADAM-9 and -10 was shown in primary prostate biopsies from patients with PCa. ADAM-1 0 was expressed in the cytoplasm and on the ceH membrane in epithelial and basal cells ofbenign prostate glands, but in high-grade PCa glands, ADAM-I 0 expression was localised to the nucleus and its expression levels appeared to be elevated when compared to low-grade PCa glands. These studies provided a strong background for the hypothesis that ADAM-9 and -10 have key roles in the development ofPCa and provided a basis for further studies.The aims of this study were to: 1) characterise the expression, localisation and levels, of ADAM-9 and -10 mRNA and protein in cell models representing characteristics of normal through androgen-dependent to androgen-independent PCa, as well as to expand the primary PCa biopsy data for ADAM-9 and ADAM-10 to encompass PCa bone metastases 2) establish an in vitro cell system, which could express elevated levels of ADAM-1 0 so that functional cell-based assays such as cell migration, invasion and attachment could be carried out, and 3) to extend the previous hormonal regulation data, to fully characterise the response of ADAM-9 and -10 mRNA and protein levels to DHT, IGF-1, DHT plus IGF-1 and EGF in the hormonal/growth factor responsive cell line LNCaP. For aim 1 (expression of ADAM-9 and -10 mRNA and protein), ADAM-9 and -10 mRNA were characterised by R T -PCR, while their protein products were analysed by Western blot. Both ADAM-9 and -10 mRNA and protein were expressed at readily detectable levels across progressively metastatic PCa cell lines model that represent characteristics of low-grade,. androgen-dependent (LNCaP and C4) to high-grade, androgen-independent (C4-2 and C4-2B) PCa. When the non-tumorigenic prostate cell line RWPE-1 was compared with the metastatic PCa cell line PC-3, differential expression patterns were seen by Western blot analysis. For ADAM-9, the active form was expressed at higher levels in RWPE-1, whilst subcellular fractionation showed that the active form of ADAM-9 was predominantly located in the cell nucleus. For ADAM-I 0, in both of the cell Jines, a nuclear specific isoform of the mature, catalytically active ADAM-I 0 was found. This isoforrn differed by -2 kDa in Mr (smaller) than the cytoplasmic specific isoform. Unprocessed ADAM-I 0 was readily detected in R WPE-1 cell lines but only occasionally detected in PC-3 cell lines. Immunocytochemistry using ADAM-9 and -10 specific antibodies confirmed nuclear, cytoplasmic and membrane expression of both ADAMs in these two cell lines. To examine the possibility of ADAM-9 and -10 being shed into the extracellular environment, membrane vesicles that are constitutively shed from the cell surface and contain membrane-associated proteins were collected from the media of the prostate cell lines RWPE-1, LNCaP and PC-3. ADAM-9 was readily detectable in RWPE- 1 and LNCaP cell membrane vesicles by Western blot analysis, but not in PC-3 cells, whilst the expression of ADAM-I 0 was detected in shed vesicles from each of these prostate cell lines. By Laser Capture Microdissection (LCM), secretory epithelial cells of primary prostate gland biopsies were isolated from benign and malignant glands. These secretory cells, by Western blot analysis, expressed similar Mr bands for ADAM-9 and -10 that were found in PCa cell lines in vitro, indicating that the nuclear specific isoforrn of ADAM-I 0 was present in PCa primary tumours and may represent the predominantly nuclear form of ADAM-I 0 expression, previously shown in high-grade PCa by immunohistochemistry (IHC). ADAM-9 and -10 were also examined by IHC in bone metastases taken from PCa patients at biopsy. Both ADAMs could be detected at levels similar to those shown for Prostate Specific Antigen (PSA) in these biopsies. Furthermore, both ADAM-9 and -10 were predominantly membrane- bound with occasional nuclear expression. For aim 2, to establish a cell system that over-expressed levels of ADAM-10, two fulllength ADAM-I 0 mammalian expression vectors were constructed; ADAM-I 0 was cloned into pcDNA3.1, which contains a CMV promoter, and into pMEP4, containing an inducible metallothionine promoter, whose activity is stimulated by the addition of CdC}z. The efficiency of these two constructs was tested by way of transient transfection in the PCa cell line PC-3, whilst the pcDNA3.1 construct was also tested in the RWPE-1 prostate cell line. Resultant Western blot analysis for all transient transfection assays showed that levels of ADAM-I 0 were not significantly elevated in any case, when compared to levels of the housekeeping gene ~-Tubulin, despite testing various levels of vector DNA, and, for pMEP4, the induction of the transfected cell system with different degrees of stimulation with CdCh to activate the metallothionine promoter post-transfection. Another study in this laboratory found similar results when the same full length ADAM-10 sequence was cloned into a Green Fluorescent Protein (GFP) expressing vector, as no fluorescence was observed by means of transient tran sfection in the same, and other, PCa cell lines. It was hypothesised that the Kozak sequence included in the full-length construct (human ADAMI 0 naturally occurring sequence) is not strong enough to initiate translation in an artificial system, in cells, which, as described in Aim 1, are already expressing readily detectable levels of endogenous ADAM-10. As a result, time constraints prevented any further progress with Aim 2 and functional studies including cell attachment, invasion and migration were unable to be explored. For Aim 3, to characterise the response of ADAM-9 and -10 mRNA and protein levels to DHT, IGF-1, DHT plus IGF-1 and EGF in LNCaP cells, the levels of ADAM-9 and -10 mRNA were not stimulated by DHT or IGF-I alone, despite our previous observations that initially characterised ADAM-9 and -10 mRNA as being responsive to DHT. However, IGF-1 in synergy with DHT did significantly elevate mRNA levels ofboth ADAMs. In the case of ADAM-9 and -10 protein, the same trends of stimulation as found at the rnRNA level were shown by Western blot analysis when ADAM-9 and -10 signal intensity was normalised with the housekeeping protein ~-Tubulin. For EGF treatment, both ADAM-9 and -10 mRNA and protein levels were significantly elevated, and further investigation vm found this to be the case for each of these ADAMs proteins in the nuclear fractions of LNCaP cells. These studies are the first to describe extensively, the expression and hormonal/growth factor regulation of two members of the ADAMs family ( -9 and -1 0) in PCa. These observations imply that the expression of ADAM-9 and -10 have varied roles in PCa whilst it develops from androgen-sensitive (early stage disease), through to an androgeninsensitive (late-stage), metastatic disease. Further studies are now required to investigate the several key areas of focus that this research has revealed, including: • Investigation of the cellular mechanisms that are involved in actively transporting the ADAMs to the cell's nuclear compartment and the ADAMs functional roles in the cell nucleus. • The construction of a full-length human ADAM-10 mammalian expression construct with the introduction of a new Kozak sequence, that elevates ADAM-I 0 expression in an in vitro cell system are required, so that functional assays such as cell invasion, migration and attachment may be carried out to fmd the functional consequences of ADAM expression on cellular behaviour. • The regulation studies also need to be extended by confirming the preliminary observations that the nuclear levels of ADAMs may also be elevated by hormones and growth factors such as DHT, IGF-1 and EGF, as well as the regulation of levels of plasma membrany vesicle associated ADAM expression. Given the data presented in this study, it is likely the ADAMs have differential roles throughout the development of PCa due to their differential cellular localisation and synergistic growth-factor regulation. These observations, along with those further studies outlined above, are necessary in identifying these specific components ofPCa metastasis to which the ADAMs may contribute.

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

Parents as coteachers of science and technology in a middle-school classroom

This study explores coteaching/cogenerative dialoguing with parents to investigate how it may be employed to engage parents more meaningfully in schools. The cogens provided a space where participants became aware of resources available for coteaching, made decisions about planning and enacting coteaching, as well as interstitial culture that facilitated positive parent-teacher relationships.

Life-cycle management of construction projects based on virtual prototyping technology

Life-cycle management (LCM) has been employed in the management of construction projects for many years in order to reduce whole life cost, time, risk and improve the service to owners. However, owing to lack of an effective information sharing platform, the current LCM of construction projects is not effectively used in the construction industry. Based upon the analysis of the information flow of LCM, a virutal prototyping (VP)-based communication and collaboration information platform is proposed. Following this, the platform is customized using DASSAULT sofware. The whole process of implementing the VP-based LCM are also discussed and, from a simple case study, it is demonstrated that the VP-based communication and collaboration information platform is an effective tool to support the LCM of construction projects.

Lift@Weimar : sustainable interaction with food, technology, and the city (Workshop)

This workshop explores innovative approaches to understanding and cultivating sustainable food culture in urban environments via human-computer-interaction (HCI) design and ubiquitous technologies. We perceive the city as an intersecting network of people, place, and technology in constant transformation. Our 2009 OZCHI workshop, Hungry 24/7? HCI Design for Sustainable Food Culture, opened a new space for discussion on this intersection amongst researchers and practitioners from diverse backgrounds including academia, government, industry, and non-for-profit organisations. Building on the past success, this new instalment of the workshop series takes a more refined view on mobile human-food interaction and the role of interactive media in engaging citizens to cultivate more sustainable everyday human-food interactions on the go. Interactive media in this sense is distributed, pervasive, and embedded in the city as a network. The workshop addresses environmental, health, and social domains of sustainability by bringing together insights across disciplines to discuss conceptual and design approaches in orchestrating mobility and interaction of people and food in the city as a network of people, place, technology, and food.

Pet food from bovine origin drying by green heat pump technology and fluidization

Bovine intestine was dried in a heat pump fluid bed combination. Minimum fluidisation velocity was calculated by Ergun Equation and some relations were established.

Urban telecommunications network : technology convergence and urban infrastructure

Rapidly developing information and telecommunication technologies and their platforms in the late 20th Century helped improve urban infrastructure management and influenced quality of life. Telecommunication technologies make it possible for people to deliver text, audio and video material using wired, wireless or fibre-optic networks. Technologies convergence amongst these digital devices continues to create new ways in which the information and telecommunication technologies are used. The 21st Century is an era where information has converged, in which people are able to access a variety of services, including internet and location based services, through multi-functional devices such as mobile phones. This chapter discusses the recent developments in telecommunication networks and trends in convergence technologies, their implications for urban infrastructure planning, and for the quality of life of urban residents.

Ubiquitous eco cities : telecommunication infrastructure, technology convergence and urban management

Efficient and effective urban management systems for Ubiquitous Eco Cities require having intelligent and integrated management mechanisms. This integration includes bringing together economic, socio-cultural and urban development with a well orchestrated, transparent and open decision-making system and necessary infrastructure and technologies. In Ubiquitous Eco Cities telecommunication technologies play an important role in monitoring and managing activities via wired and wireless networks. Particularly, technology convergence creates new ways in which information and telecommunication technologies are used and formed the backbone of urban management. The 21st Century is an era where information has converged, in which people are able to access a variety of services, including internet and location based services, through multi-functional devices and provides new opportunities in the management of Ubiquitous Eco Cities. This chapter discusses developments in telecommunication infrastructure and trends in convergence technologies and their implications on the management of Ubiquitous Eco Cities

Self-service technology and the moderating effects of consumer characteristics

The incidence of self-service technology, where the consumer delivers the service themselves using technology, is increasing in the service encounter. One area that is under-explored is the potential impact of self-service technology on consumer satisfaction and affective commitment. Accordingly, this paper presents an empirical study that investigates the relative impact of self-service technology on consumer satisfaction (both overall and transaction-specific) and affective commitment, accounting for the moderating effects of consumer characteristics. The results highlight the importance of personal service for evaluations of satisfaction and commitment, and the importance of social competency as a moderator in this relationship. An understanding of these consumer perceptions will allow organisations to develop strategies to deliver the services expected by their consumers, improving consumer satisfaction and commitment.

Ubiquitous eco cities : infrastructure, technology and management

Efficient and effective urban management systems for Ubiquitous Eco Cities require having intelligent and integrated management mechanisms. This integration includes bringing together economic, socio-cultural and urban development with a well orchestrated, transparent and open decision-making system and necessary infrastructure and technologies. In Ubiquitous Eco Cities telecommunication technologies play an important role in monitoring and managing activities via wired and wireless networks. Particularly, technology convergence creates new ways in which information and telecommunication technologies are used and formed the backbone of urban management. The 21st Century is an era where information has converged, in which people are able to access a variety of services, including internet and location based services, through multi-functional devices and provides new opportunities in the management of Ubiquitous Eco Cities. This chapter discusses developments in telecommunication infrastructure and trends in convergence technologies and their implications on the management of Ubiquitous Eco Cities.

meso-Porphyrinylphosphine Oxides: Mono- and Bidentate Ligands for Supramolecular Chemistry and the Crystal Structures of Monomeric [10,20-Diphenylporphyrinatonickel(II)-5,15-diyl]-bis-[P(O)Ph2] and Polymeric Self-Coordinated ([10,20-Diphenylporphyrinatozinc(II)-5,15-diyl]-bis-[P(O)Ph2])

A series of porphyrins substituted in one or two meso-positions by diphenylphosphine oxide groups has been prepared by the palladium catalysed reaction of diphenylphosphine or its oxide with the corresponding bromoporphyrins. Compounds {MDPP-[P(O)Ph2]n} (M = H2, Ni, Zn; H2DPP = 5,15-diphenylporphyrin; n = 1, 2) were isolated in yields of 60-95%. The reaction is believed to proceed via the conventional oxidative addition, phosphination and reductive elimination steps, as the stoichiometric reaction of η1-palladio(II) porphyrin [PdBr(H2DPP)(dppe)] (H2DPP = 5,15-diphenylporphyrin; dppe = 1,2-bis(diphenylphosphino)ethane) with diphenylphosphine oxide also results in the desired mono-porphyrinylphosphine oxide [H2DPP-P(O)Ph2]. Attempts to isolate the tertiary phosphines failed due to their extreme air-sensitivity. Variable temperature 1H NMR studies of [H2DPP-P(O)Ph2] revealed an intrinsic lack of symmetry, while fluorescence spectroscopy showed that the phosphine oxide group does not behave as a "heavy atom" quencher. The electron withdrawing effect of the phosphine oxide group was confirmed by voltammetry. The ligands were characterised by multinuclear NMR and UV-visible spectroscopy as well as mass spectrometry. Single crystal X-ray crystallography showed that the bis(phosphine oxide) nickel(II) complex {[NiDPP-[P(O)Ph2]2} is monomeric in the solid state, with a ruffled porphyrin core and the two P=O fragments on the same side of the average plane of the molecule. On the other hand, the corresponding zinc(II) complex formed infinite chains through coordination of one Ph2PO substituent to the neighbouring zinc porphyrin through an almost linear P=O---Zn unit, leaving the other Ph2PO group facing into a parallel channel filled with disordered water molecules. These new phosphine oxides are attractive ligands for supramolecular porphyrin chemistry.

Ureaplasma parvum and ureaplasma urealyticum detected in semen after washing prior to assisted reproductive technology procedures

Objective: This study investigated: (i) the prevalence of ureaplasmas in semen and washed semen and (ii) the effect of ureaplasmas on semen andrology parameters. Design: Prospective study. Setting: IVF unit -private hospital, Brisbane, Australia. Patient(s): Three hundred and forty three men participating in an assisted reproductive technology (ART) treatment cycle. Intervention(s): Semen and washed semen tested by culture, PCR assays and indirect immunofluorescent antibody assays. Statistical differences were determined by a t-test, Wilcoxon or Pearson’s Chi- square test where appropriate. Main Outcome Measure(s): The prevalence of ureaplasmas in semen and washed semen and the effect of these microorganisms on semen andrology parameters. Result(s): Ureaplasmas were detected in 73/343 (22%) semen samples and 29/343 (8.5%) washed semen samples. Ureaplasmas adherent to the surface of spermatozoa were demonstrated by indirect immunofluorescent antibody testing. U. parvum serovar 6 (36.6%) and U. urealyticum (30%) were the most prevalent isolates in washed semen. A comparison of the semen andrology parameters of washed semen ureaplasma positive and negative groups demonstrated a lower proportion of non-motile sperm in the washed semen ureaplasma positive group. Conclusion(s): Ureaplasmas are not always removed from semen by a standard ART washing procedure and can remain adherent to the surface of spermatozoa.

Technology strategy and competitive performance in bridge construction

Pragmatic construction professionals, accustomed to intense price competition and focused on the bottom line, have difficulty justifying investments in advanced technology. Researchers and industry professionals need improved tools to analyze how technology affects the performance of the firm. This paper reports the results of research to begin answering the question, “does technology matter?” The researchers developed a set of five dimensions for technology strategy, collected information regarding these dimensions along with four measures of competitive performance in five bridge construction firms, and analyzed the information to identify relationships between technology strategy and competitive performance. Three technology strategy dimensions—competitive positioning, depth of technology strategy, and organizational fit—showed particularly strong correlations with the competitive performance indicators of absolute growth in contract awards and contract award value per technical employee. These findings indicate that technology does matter. The research also provides ways to analyze options for approaching technology and ways to relate technology to competitive performance for use by managers. It also provides a valuable set of research measures for technology strategy.

Agency, tangible technology and young children

This paper explores the embodiment of agency concepts in tangible user interfaces to create meaningful learning experiences. Current notions of agent-based tangible technology are extended, through the development of low-fidelity prototypes, to include additional flexibility and adaptability. A study involving these prototypes was conducted in a kindergarten environment with nine four-year-old children. Observations of children's interactions with the prototypes produced insightful results which will be used to further refine the product under development.