Modulation of the SHBG signalling axis

Sex hormone-binding globulin (SHBG) is a homodimeric plasma glycoprotein that is the major sex steroid carrier-protein in the bloodstream and functions also as a key regulator of steroid bioavailability within target tissues, such as the prostate. Additionally, SHBG binds to prostatic cell membranes via the putative and unidentified SHBG receptor (RSHBG), activating a signal transduction pathway implicated in stimulating both proliferation and expression of prostate specific antigen (PSA) in prostate cell lines in vitro. A yeast-two hybrid assay suggested an interaction between SHBG and kallikrein-related protease (KLK) 4, which is a serine protease implicated in the progression of prostate cancer. The potential interaction between these two proteins was investigated in this PhD thesis to determine whether SHBG is a proteolytic substrate of KLK4 and other members of the KLK family including KLK3/PSA, KLK7 and KLK14. Furthermore, the effects from SHBG proteolytic degradation on SHBG-regulated steroid bioavailability and the activation of the putative RSHBG signal transduction pathway were examined in the LNCaP prostate cancer cell line. SHBG was found to be a proteolytic substrate of the trypsin-like KLK4 and KLK14 in vitro, yielding several proteolysis fragments. Both chymotrypsin-like PSA and KLK7 displayed insignificant proteolytic activity against SHBG. The kinetic parameters of SHBG proteolysis by KLK4 and KLK14 demonstrate a strong enzyme-substrate binding capacity, possessing a Km of 1.2 ± 0.7 µM and 2.1 ± 0.6 µM respectively. The catalytic efficiencies (kcat/Km) of KLK4 and KLK14 proteolysis of SHBG were 1.6 x 104 M-1s-1 and 3.8 x 104 M-1s-1 respectively, which were comparable to parameters previously reported for peptide substrates. N-terminal sequencing of the fragments revealed cleavage near the junction of the N- and C-terminal laminin globulin-like (G-like) domains of SHBG, resulting in the division of the two globulins and ultimately the full degradation of these fragments by KLK4 and KLK14 over time. Proteolytic fragments that may retain steroid binding were rapidly degraded by both proteases, while fragments containing residues beyond the steroid binding pocket were less degraded over the same period of time. Degradation of SHBG was inhibited by the divalent metal cations calcium and zinc for KLK4, and calcium, zinc and magnesium for KLK14. The human secreted serine protease inhibitors (serpins), α1-antitrypsin and α2-antiplasmin, inhibited KLK4 and KLK14 proteolysis of SHBG; α1-antichymotrypsin inhibited KLK4 but not KLK14 activity. The inhibition by these serpins was comparable and in some cases more effective than general trypsin protease inhibitors such as aprotinin and phenylmethanesulfonyl fluoride (PMSF). The binding of 5α-dihydrotestosterone (DHT) to SHBG modulated interactions with KLK4 and KLK14. Steroid-free SHBG was more readily digested by both enzymes than DHT-bound SHBG. Moreover, a binding interaction exists between SHBG and pro-KLK4 and pro-KLK14, with DHT strengthening the binding to pro-KLK4 only. The inhibition of androgen uptake by cultured prostate cancer cells, mediated by SHBG steroid-binding, was examined to assess whether SHBG proteolysis by KLK4 and KLK14 modulated this process. Proteolytic digestion eliminated the ability of SHBG to inhibit the uptake of DHT from conditioned media into LNCaP cells. Therefore, the proteolysis of SHBG by KLK4 and KLK14 increased steroid bioavailability in vitro, leading to an increased uptake of androgens by prostate cancer cells. Interestingly, different transcriptional responses of PSA and KLK2, which are androgen-regulated genes, to DHT-bounsd SHBG treatment were observed between low and high passage number LNCaP cells (lpLNCaP and hpLNCaP respectively). HpLNCaP cells treated with DHT-bound SHBG demonstrated a significant synergistic upregulation of PSA and KLK2 above DHT or SHBG treatment alone, which is similar to previously reported downstream responses from RSHBG-mediated signaling activation. As this result was not seen in lpLNCaP cells, only hpLNCaP cells were further investigated to examine the modulation of potential RSHBG activity by KLK4 and KLK14 proteolysis of SHBG. Contrary to reported results, no increase in intracellular cAMP was observed in hpLNCaP cells when treated with SHBG in the presence and absence of either DHT or estradiol. As a result, the modulation of RSHBG-mediated signaling activation could not be determined. Finally, the identification of the RSHBG from both breast (MCF-7) and prostate cancer (LNCaP) cell lines was attempted. Fluorescently labeled peptides corresponding to the putative receptor binding domain (RBD) of SHBG were shown to be internalized by MCF-7 cells. Crosslinking of the RBD peptide to the cell surfaces of both MCF-7 and LNCaP cells, demonstrated the interaction of the peptide with several targets. These targets were then captured using RBD peptides synthesized onto a hydrophilic scaffold and analysed by mass spectrometry. The samples captured by the RBD peptide returned statistically significantly matches for cytokeratin 8, 18 and 19 as well as microtubule-actin crosslinking factor 1, which may indicate a novel interaction between SHBG and these proteins, but ultimately failed to detect a membrane receptor potentially responsible for the putative RSHBG-mediated signaling. This PhD project has reported the proteolytic processing of SHBG by two members of the kallikrein family, KLK4 and KLK14. The effect of SHBG proteolysis by KLK4 and KLK14 on RSHBG-mediated signaling activation was unable to be determined as the reported signal transduction pathway was not activated after treatment with SHBG, in combination with either DHT or estradiol. However, the digestion of SHBG by these two proteases positively regulated androgen bioavailability to prostate cancer cells in vitro. The increased uptake of androgens is deleterious in prostate cancer due to the promotion of proliferation, metastasis, invasion and the inhibition of apoptosis. The increased bioavailability of androgens, from SHBG proteolysis by KLK4 and KLK14, may therefore promote both carcinogenesis and progression of prostate cancer. Finally, this information may contribute to the development of therapeutic treatment strategies for prostate cancer by inhibiting the proteolysis of SHBG, by KLK4 and KLK14, to prevent the increased uptake of androgens by hormone-dependent cancerous tissues.

Revisiting the conceptualization and measurement of service quality

Since the 1980s, industries and researchers have sought to better understand the quality of services due to the rise in their importance (Brogowicz, Delene and Lyth 1990). More recent developments with online services, coupled with growing recognition of service quality (SQ) as a key contributor to national economies and as an increasingly important competitive differentiator, amplify the need to revisit our understanding of SQ and its measurement. Although ‘SQ’ can be broadly defined as “a global overarching judgment or attitude relating to the overall excellence or superiority of a service” (Parasuraman, Berry and Zeithaml 1988), the term has many interpretations. There has been considerable progress on how to measure SQ perceptions, but little consensus has been achieved on what should be measured. There is agreement that SQ is multi-dimensional, but little agreement as to the nature or content of these dimensions (Brady and Cronin 2001). For example, within the banking sector, there exist multiple SQ models, each consisting of varying dimensions. The existence of multiple conceptions and the lack of a unifying theory bring the credibility of existing conceptions into question, and beg the question of whether it is possible at some higher level to define SQ broadly such that it spans all service types and industries. This research aims to explore the viability of a universal conception of SQ, primarily through a careful re-visitation of the services and SQ literature. The study analyses the strengths and weaknesses of the highly regarded and widely used global SQ model (SERVQUAL) which reflects a single-level approach to SQ measurement. The SERVQUAL model states that customers evaluate SQ (of each service encounter) based on five dimensions namely reliability, assurance, tangibles, empathy and responsibility. SERVQUAL, however, failed to address what needs to be reliable, assured, tangible, empathetic and responsible. This research also addresses a more recent global SQ model from Brady and Cronin (2001); the B&C (2001) model, that has potential to be the successor of SERVQUAL in that it encompasses other global SQ models and addresses the ‘what’ questions that SERVQUAL didn’t. The B&C (2001) model conceives SQ as being multidimensional and multi-level; this hierarchical approach to SQ measurement better reflecting human perceptions. In-line with the initial intention of SERVQUAL, which was developed to be generalizable across industries and service types, this research aims to develop a conceptual understanding of SQ, via literature and reflection, that encompasses the content/nature of factors related to SQ; and addresses the benefits and weaknesses of various SQ measurement approaches (i.e. disconfirmation versus perceptions-only). Such understanding of SQ seeks to transcend industries and service types with the intention of extending our knowledge of SQ and assisting practitioners in understanding and evaluating SQ. The candidate’s research has been conducted within, and seeks to contribute to, the ‘IS-Impact’ research track of the IT Professional Services (ITPS) Research Program at QUT. The vision of the track is “to develop the most widely employed model for benchmarking Information Systems in organizations for the joint benefit of research and practice.” The ‘IS-Impact’ research track has developed an Information Systems (IS) success measurement model, the IS-Impact Model (Gable, Sedera and Chan 2008), which seeks to fulfill the track’s vision. Results of this study will help future researchers in the ‘IS-Impact’ research track address questions such as: • Is SQ an antecedent or consequence of the IS-Impact model or both? • Has SQ already been addressed by existing measures of the IS-Impact model? • Is SQ a separate, new dimension of the IS-Impact model? • Is SQ an alternative conception of the IS? Results from the candidate’s research suggest that SQ dimensions can be classified at a higher level which is encompassed by the B&C (2001) model’s 3 primary dimensions (interaction, physical environment and outcome). The candidate also notes that it might be viable to re-word the ‘physical environment quality’ primary dimension to ‘environment quality’ so as to better encompass both physical and virtual scenarios (E.g: web sites). The candidate does not rule out the global feasibility of the B&C (2001) model’s nine sub-dimensions, however, acknowledges that more work has to be done to better define the sub-dimensions. The candidate observes that the ‘expertise’, ‘design’ and ‘valence’ sub-dimensions are supportive representations of the ‘interaction’, physical environment’ and ‘outcome’ primary dimensions respectively. The latter statement suggests that customers evaluate each primary dimension (or each higher level of SQ classification) namely ‘interaction’, physical environment’ and ‘outcome’ based on the ‘expertise’, ‘design’ and ‘valence’ sub-dimensions respectively. The ability to classify SQ dimensions at a higher level coupled with support for the measures that make up this higher level, leads the candidate to propose the B&C (2001) model as a unifying theory that acts as a starting point to measuring SQ and the SQ of IS. The candidate also notes, in parallel with the continuing validation and generalization of the IS-Impact model, that there is value in alternatively conceptualizing the IS as a ‘service’ and ultimately triangulating measures of IS SQ with the IS-Impact model. These further efforts are beyond the scope of the candidate’s study. Results from the candidate’s research also suggest that both the disconfirmation and perceptions-only approaches have their merits and the choice of approach would depend on the objective(s) of the study. Should the objective(s) be an overall evaluation of SQ, the perceptions-only approached is more appropriate as this approach is more straightforward and reduces administrative overheads in the process. However, should the objective(s) be to identify SQ gaps (shortfalls), the (measured) disconfirmation approach is more appropriate as this approach has the ability to identify areas that need improvement.

The interaction order of Second Life : how micro sociology can contribute to online games innovation

This paper uses the virtual world Second Life (as Web 2.0 environment) to discuss how sociological theory is a relevant tool for innovation in the area of games design as a methodological strategy. Via the theories of Erving Goffman’s interaction order the paper illustrates how micro studies of online interaction demonstrate active accounts of membership and complex interactivity. In order to achieve this, the paper outlines a methodological tool to assist in the application of micro sociology to Web 2.0 environments that accounts for the multiple dimensions of participation within the digital field.

Multi-vehicles interaction graph model for cooperative collision warning system

Cooperative collision warning system for road vehicles, enabled by recent advances in positioning systems and wireless communication technologies, can potentially reduce traffic accident significantly. To improve the system, we propose a graph model to represent interactions between multiple road vehicles in a specific region and at a specific time. Given a list of vehicles in vicinity, we can generate the interaction graph using several rules that consider vehicle's properties such as position, speed, heading, etc. Safety applications can use the model to improve emergency warning accuracy and optimize wireless channel usage. The model allows us to develop some congestion control strategies for an efficient multi-hop broadcast protocol.

Media scepticism, media diets and media landscapes: A consideration of US versus Australian political information environments

The range of political information sources available to modern Australians is greater and more varied today than at any point in the nation’s history, incorporating print, broadcast, Internet, mainstream and non-mainstream media. In such a competitive media environment, the factors which influence the selection of some information sources above others are of interest to political agents, media institutions and communications researchers alike. A key factor in information source selection is credibility. At the same time that the range of political information sources is increasing rapidly, due to the development of new information and communication technologies, audience research suggests that trust in mainstream media organisations in many countries is declining. So if people distrust the mainstream media, but have a vast array of alternative political information sources available to them, what do their personal media consumption patterns look like? How can we analyse such media consumption patterns in a meaningful way? In this paper I will briefly map the development of media credibility research in the US and Australia, leading to a discussion of one of the most recent media credibility constructs to be shown to influence political information consumption, media scepticism. Looking at the consequences of media scepticism, I will then consider the associated media consumption construct, media diet, and evaluate its usefulness in an Australian, as opposed to US, context. Finally, I will suggest alternative conceptualisations of media diets which may be more suited to Australian political communications research.

"I'm thinking 'bout moving in"... An examination of the issues impacting on the recruitment of secondary school graduates to the library and information profession

In recent years concern has been expressed internationally about the future of the library and information services (LIS) profession: recruitment and retention, changing skill sets and declining numbers of people choosing librarianship as a career are all factors contributing to an uncertain future. One area yet explored in any depth is the topic of why LIS studies are not perceived, let alone promoted, as a good first professional qualification for high school graduates. This paper considers the professional literature that examines the uptake of librarianship as a first qualification by school leavers and discusses, in the context of the Australian library sector, the role of professional associations, library schools, National and State Libraries, as well as individual libraries and librarians. Examples of best practice are presented to highlight the opportunities for inspiring and motivating students through well structured and stimulating work experience programs. The topic is relevant to all librarians who are interested in the future of the LIS profession. It is argued that the focus of the present conference on ‘moving up’ and ‘moving on’ can only have real significance when the profession has a more complete understanding of the barriers to and the opportunities for ‘moving in’.

Aligning policy with practice: an evaluation of vocational education and training in the library and information services sector

Vocational education and training for the library and information services (LIS) sector in Australia offers students the career pathway to become library technicians. Library technicians play a valuable role in drawing on sound practical knowledge and skills to support the delivery of library and information services that meet client needs. Over the past forty years, the Australian Library and Information Association (ALIA) has monitored the quality of library technician courses. Since 2005, ALIA has run national professional development days for library technician educators with the goal of establishing an alternative model for course recognition focusing on the process of peer review to benchmark good practice and stimulate continuous improvement in library technician education. This initial developmental work has culminated in 2009 with site visits to all library technician courses in Australia. The paper presents a whole-of-industry case study to critically review the work undertaken to date.

The strategic and operational dimensions of staff training and professional development for information professionals : what neXus2 has revealed

The paper seeks to continue the debate about the need for professionals in the library and information services (LIS) sector to continually engage in career-long learning to sustain and develop their knowledge and skills in a dynamic industry. Aims: The neXus2 workforce study has been funded by the ALIA and the consortium of National and State Libraries Australasia (NSLA). It builds on earlier research work (the neXus census) that looked at the demographic, educational and career perspectives of individual library and information professions, to critically examine institutional policies and practices associated with the LIS workforce. The research aims to develop a clearer understanding of the issues impacting on workforce sustainability, workforce capability and workforce optimisation. Methods: The research methodology involved an extensive online survey conducted in March 2008 which collected data on organisational and general staffing; recruitment and retention; staff development and continuing professional education; and succession planning. Encouragement to participate was provided by key industry groups, including academic, public, health, law and government library and information agencies, with the result that around 150 institutions completed the questionnaire. Results: The paper will specifically discuss the research findings relating to training and professional development, to measure the scope and distribution of training activities across the workforce, to consider the interrelationship between the strategic and operational dimensions of staff development in individual institutions and to analyse the common and distinctive factors evident in the different sectors of the profession. Conclusion: The neXus2 project has successfully engaged LIS institutions in the collection of complex industry data that is relevant to the future education and workforce strategies for all areas of the profession. Cross-sector forums such as Information Online 2009 offer the opportunity for stimulating professional dialogue on the key issues.

Correlating eye gaze direction, depth and vehicle information on an interactive map for driver training

The over represented number of novice drivers involved in crashes is alarming. Driver training is one of the interventions aimed at mitigating the number of crashes that involve young drivers. Experienced drivers have better hazard perception ability compared to inexperienced drivers. Eye gaze patterns have been found to be an indicator of the driver's competency level. The aim of this paper is to develop an in-vehicle system which correlates information about the driver's gaze and vehicle dynamics, which is then used to assist driver trainers in assessing driving competency. This system allows visualization of the complete driving manoeuvre data on interactive maps. It uses an eye tracker and perspective projection algorithms to compute the depth of gaze and plots it on Google maps. This interactive map also features the trajectory of the vehicle and turn indicator usage. This system allows efficient and user friendly analysis of the driving task. It can be used by driver trainers and trainees to understand objectively the risks encountered during driving manoeuvres. This paper presents a prototype that plots the driver's eye gaze depth and direction on an interactive map along with the vehicle dynamics information. This prototype will be used in future to study the difference in gaze patterns in novice and experienced drivers prior to a certain manoeuvre.

The Brisbane media map : participatory design and authentic learning to link students and industry

The Brisbane Media Map is both an online resource and a tertiary-level authentic learning project. The Brisbane Media Map is an online database which provides a detailed overview of about 600 media industry organisations in Brisbane, Australia. In addition to providing contact details and synopses for each organisation’s profile, the Brisbane Media Map also includes supplementary information on current issues, trends, and individuals in the media and communication industry sectors. This resource is produced and updated annually by final-year undergraduate Media and Communication students. This article introduces the Brisbane Media Map, its functionality and systems design approach, as well as its alignment with key learning infrastructures. It examines authentic learning as the pedagogical framework underpinning the ongoing development work of the resource and highlights some synergies of this framework with participatory design principles. The Brisbane Media Map is a useful example of an authentic learning approach that successfully engages students of non-traditional and non-design areas of study in human-computer interaction, usability, and participatory design activities.

Increasing the Information available to Coroners : the effect on autopsy decision making

This paper details research completed in 2007 which investigated autopsy decision making in a death investigation. The data was gathered during the first year of operation of a new Coroners Act in Queensland, Australia, which changed the process of death investigation in three ways which are important to this paper. First, it required a greater amount of information to be gathered at the scene by police, and this included a thorough investigation of the circumstances of the death, including statements from witnesses, friends and family, as well as evidence gathering at the scene. Second, it required Coroners, for the first time, to determine the level of invasiveness of the autopsy required to complete the death investigation. Third, it enabled the communication of a genuine family concern, to be communicated to the Coroner. The outcome of such information was threefold. First, a greater amount of information offered to the Coroner led to a decrease in the number of full internal autopsies ordered, but an increase in the number of partial internal autopsies ordered. Second, this shift in autopsy decision making by Coroners saw certain factors given greater importance than others in decisions to order full internal or external only autopsies. Third, a raised family concern had a significant impact on autopsy decision making and tended to decrease the invasiveness of the autopsy ordered by Coroners.

Patterns of query reformulation during web searching

Query reformulation is a key user behavior during Web search. Our research goal is to develop predictive models of query reformulation during Web searching. This article reports results from a study in which we automatically classified the query-reformulation patterns for 964,780 Web searching sessions, composed of 1,523,072 queries, to predict the next query reformulation. We employed an n-gram modeling approach to describe the probability of users transitioning from one query-reformulation state to another to predict their next state. We developed first-, second-, third-, and fourth-order models and evaluated each model for accuracy of prediction, coverage of the dataset, and complexity of the possible pattern set. The results show that Reformulation and Assistance account for approximately 45% of all query reformulations; furthermore, the results demonstrate that the first- and second-order models provide the best predictability, between 28 and 40% overall and higher than 70% for some patterns. Implications are that the n-gram approach can be used for improving searching systems and searching assistance.

Small molecule–biopolymer interactions : Ultraviolet–visible and fluorescence spectroscopy and chemometrics

Interactions between small molecules with biopolymers e.g. the bovine serum albumin (BSA protein), are important, and significant information is recorded in the UV–vis and fluorescence spectra of their reaction mixtures. The extraction of this information is difficult conventionally and principally because there is significant overlapping of the spectra of the three analytes in the mixture. The interaction of berberine chloride (BC) and the BSA protein provides an interesting example of such complex systems. UV–vis and fluorescence spectra of BC and BSA mixtures were investigated in pH 7.4 Tris–HCl buffer at 37 °C. Two sample series were measured by each technique: (1) [BSA] was kept constant and the [BC] was varied and (2) [BC] was kept constant and the [BSA] was varied. This produced four spectral data matrices, which were combined into one expanded spectral matrix. This was processed by the multivariate curve resolution–alternating least squares method (MCR–ALS). The results produced: (1) the extracted pure BC, BSA and the BC–BSA complex spectra from the measured heavily overlapping composite responses, (2) the concentration profiles of BC, BSA and the BC–BSA complex, which are difficult to obtain by conventional means, and (3) estimates of the number of binding sites of BC.

Spectrometric and voltammetric studies of the interaction between quercetin and bovine serum albumin using warfarin as site marker with the aid of chemometrics

The interaction of quercetin, which is a bioflavonoid, with bovine serum albumin (BSA) was investigated under pseudo-physiological conditions by the application of UV–vis spectrometry, spectrofluorimetry and cyclic voltammetry (CV). These studies indicated a cooperative interaction between the quercetin–BSA complex and warfarin, which produced a ternary complex, quercetin–BSA–warfarin. It was found that both quercetin and warfarin were located in site I. However, the spectra of these three components overlapped and the chemometrics method – multivariate curve resolution-alternating least squares (MCR-ALS) was applied to resolve the spectra. The resolved spectra of quercetin–BSA and warfarin agreed well with their measured spectra, and importantly, the spectrum of the quercetin–BSA–warfarin complex was extracted. These results allowed the rationalization of the behaviour of the overlapping spectra. At lower concentrations ([warfarin] < 1 × 10−5 mol L−1), most of the site marker reacted with the quercetin–BSA, but free warfarin was present at higher concentrations. Interestingly, the ratio between quercetin–BSA and warfarin was found to be 1:2, suggesting a quercetin–BSA–(warfarin)2 complex, and the estimated equilibrium constant was 1.4 × 1011 M−2. The results suggest that at low concentrations, warfarin binds at the high-affinity sites (HAS), while low-affinity binding sites (LAS) are occupied at higher concentrations.

neXus2. An investigation into the library and information services workforce in Australia: The institutional perspective. Final report prepared for the Australian Library and Information Association and National and State Libraries Australia

The neXus2 research project has sought to investigate the library and information services (LIS) workforce in Australia, from the institutional or employer perspective. The study builds on the neXus1 study, which collected data from individuals in the LIS workforce in order to present a snapshot of the profession in 2006, highlighting the demographics, educational background and career details of library and information professionals in Australia. To counterbalance this individual perspective, library institutions were invited to participate in a survey to contribute further data as employers. This final report on the neXus2 project compares the findings from the different library sectors, ie academic libraries, TAFE libraries, the National and State libraries, public libraries, special libraries and school libraries.