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.

Departing from the virtual community : a discussion of the ways and means to uncover localised order in online games

This paper is part one of a three part study into the collective regulation processes of players in massive multiplayer online games (MMOG). Traditionally game playing has not been classed as problematic, however with introduction of new media technologies and new ways to play games, certain contexts have become obscure, namely the localised order of ‘playing online’ or how players manage and maintain order between each other as opposed to ‘following the rules’. Principally this paper will examine concepts of ‘virtual community’. These will be illustrated as particularly unhelpful when considering how people conduct themselves in these spaces. Thus, ‘virtual community’ will be seen as critical in implicating various online behaviours as superior to other online behaviours causing obscurity and blurring actions. This obscurity is grounded by strong associations in the virtual community as logic of practise in and of itself; behaviours that fall outside this category become common sense and as such are made invisible for investigation. This paper will draw upon the theories of Basil Bernstein and Pierre Bourdieu to produce a distinction between online behaviours and ultimately make them visible for further investigation. In doing so this paper seeks to form a basis for future research where interaction in these spaces can be identified as belonging to a certain framework to inform the design of online games and applications more effectively.

Psychotherapy in Australia : clinical psychology and its approach to depression

In Australia, clinical psychology training is dominated by cognitive and behavioral treatments (CBTs), although there is exposure to other theoretical orientations. Since 2001, over 20% of general medical practitioners (GPs) have received training in CBT, and psychiatry training increasingly incorporates CBT elements. Psychotherapy by medical practitioners is financially supported by universal health care funding with supplementation by patients and their private health insurance. Federally funded health benefits for up to 12 psychology consultations per year are provided on referral from GPs and psychiatrists, and initial take up has been very strong. Mrs. A would be a typical patient for such a referral. However, she would not fulfil criteria for priority access from state-funded mental health services. Mrs. A would probably consult a GP and receive antidepressants, although she may also access a range of other community support programs. Access to and acceptance of psychotherapy would be greater in urban areas, and if she were of Anglo-Saxon and non- indigenous origin.

Challenges in implementing evidence-based practice into mental health services

This paper highlights challenges in implementing mental health policy at a service delivery level. It describes an attempt to foster greater application of recovery-orientated principles and practices within mental health services. Notwithstanding a highly supportive policy environment, strong support from service administrators, and an enthusiastic staff response to training, application of the training and support tools was weaker than anticipated. This paper evaluates the dissemination trial against key elements to promote sustained adoption of innovations. Organisational and procedural changes are required before mental health policies are systematically implemented in practice.

Automating human thought processes for a UAV forced landing

This paper describes the current status of a program to develop an automated forced landing system for a fixed-wing Unmanned Aerial Vehicle (UAV). This automated system seeks to emulate human pilot thought processes when planning for and conducting an engine-off emergency landing. Firstly, a path planning algorithm that extends Dubins curves to 3D space is presented. This planning element is then combined with a nonlinear guidance and control logic, and simulated test results demonstrate the robustness of this approach to strong winds during a glided descent. The average path deviation errors incurred are comparable to or even better than that of manned, powered aircraft. Secondly, a study into suitable multi-criteria decision making approaches and the problems that confront the decision-maker is presented. From this study, it is believed that decision processes that utilize human expert knowledge and fuzzy logic reasoning are most suited to the problem at hand, and further investigations will be conducted to identify the particular technique/s to be implemented in simulations and field tests. The automated UAV forced landing approach presented in this paper is promising, and will allow the progression of this technology from the development and simulation stages through to a prototype system

Imaginary relish and exquisite torture : the elaborated intrusion theory of desire

The authors argue that human desire involves conscious cognition that has strong affective connotation and is potentially involved in the determination of appetitive behavior rather than being epiphenomenal to it. Intrusive thoughts about appetitive targets are triggered automatically by external or physiological cues and by cognitive associates. When intrusions elicit significant pleasure or relief, cognitive elaboration usually ensues. Elaboration competes with concurrent cognitive tasks through retrieval of target related information and its retention in working memory. Sensory images are especially important products of intrusion and elaboration because they simulate the sensory and emotional qualities of target acquisition. Desire images are momentarily rewarding but amplify awareness of somatic and emotional deficits. Effects of desires on behavior are moderated by competing incentives, target availability, and skills. The theory provides a coherent account of existing data and suggests new directions for research and treatment.

Innovative approaches to intervention for problem drinking

Purpose of review: To critique the recent literature on telephone, correspondence-based, and computerized interventions for alcohol problems, which enhance or substitute for practitioner-delivered treatments. Recent findings: There is an unmet need for screening, assessment and intervention for alcohol problems, in part because of the difficulty in accessing such treatment within the current health care system. Research on the efficacy of correspondence or electronic (for example Internet-based) interventions is beginning to emerge. In the period 2003–2004 we identified nine acceptability or feasibility studies of these approaches and seven efficacy trials covering a wide range of settings. These modes of intervention are acceptable to patients and the public, and with careful planning, can be implemented in a variety of settings. Treatment trials demonstrate the efficacy of these interventions in reducing hazardous drinking by university students, in delaying initiation of heavy drinking in children and adolescents, and, intriguingly, in addressing insomnia among recovering alcoholics. Summary: There is strong support among potential users for alcohol interventions that employ telephone assistance, written correspondence, and the Internet. These new technologies offer the prospect of increasing the reach of interventions for problem drinking and being cost- effective alternatives or supplements to face-to-face health service delivery.

Supervision practices in allied mental health : relationships of supervision characteristics to perceived impact and job satisfaction

This study examined whether supervision characteristics impacted on mental health practice and morale, and developed a new Supervision Attitude Scale (SAS). Telephone surveys were conducted with a representative sample of 272 staff from public mental health services across Queensland. Although supervision was widely received and positively rated, it had low average intensity, and assessment and training of skills was rarely incorporated. Perceived impact on practice was associated with acquisition of skills and positive attitudes to supervisors, but extent of supervision was related to impact only if it was from within the profession. Intention to resign was unrelated to extent of supervision, but was associated with positive attitudes to supervisors, accessibility, high impact, and empathy or praise in supervision sessions. The SAS had high internal consistency, and its intercorrelations were consistent with it being a measure of relationship positivity. The study supported the role of supervision in retention and in improving practice. It also highlighted supervision characteristics that might be targeted in training, and provided preliminary data on a new measure.

Corneal sensitivity and slit scanning in vivo confocal microscopy of the subbasal nerve plexus of the normal central and peripheral human cornea

Purpose: To determine the subbasal nerve density and tortuosity at 5 corneal locations and to investigate whether these microstructural observations correlate with corneal sensitivity. Method: Sixty eyes of 60 normal human subjects were recruited into 1 of 3 age groups, group 1: aged ,35 years, group 2: aged 35–50 years, and group 3: aged .50 years. All eyes were examined using slit-lamp biomicroscopy, noncontact corneal esthesiometry, and slit scanning in vivo confocal microscopy. Results: The mean subbasal nerve density and the mean corneal sensitivity were greatest centrally (14,731 6 6056 mm/mm2 and 0.38 6 0.21 millibars, respectively) and lowest in the nasal mid periphery (7850 6 4947 mm/mm2 and 0.49 6 0.25 millibars, respectively). The mean subbasal nerve tortuosity coefficient was greatest in the temporal mid periphery (27.3 6 6.4) and lowest in the superior mid periphery (19.3 6 14.1). There was no significant difference in mean total subbasal nerve density between age groups. However, corneal sensation (P = 0.001) and subbasal nerve tortuosity (P = 0.004) demonstrated significant differences between age groups. Subbasal nerve density only showed significant correlations with corneal sensitivity threshold in the temporal cornea and with subbasal nerve tortuosity in the inferior and nasal cornea. However, these correlations were weak. Conclusions: This study quantitatively analyzes living human corneal nerve structure and an aspect of nerve function. There is no strong correlation between subbasal nerve density and corneal sensation. This study provides useful baseline data for the normal living human cornea at central and mid-peripheral locations

The studio sans frontières : the open-studio as an answer to the challenges of teaching contemporary art

Since at least the 1960s, art has assumed a breadth of form and medium as diverse as social reality itself. Where once it was marginal and transgressive for artists to work across a spectrum of media, today it is common practice. In this ‘post-medium’ age, fidelity to a specific branch of media is a matter of preference, rather than a code of practice policed by gallerists, curators and critics. Despite the openness of contemporary art practice, the teaching of art at most universities remains steadfastly discipline-based. Discipline-based art teaching, while offering the promise of focussed ‘mastery’ of a particular set of technical skills and theoretical concerns, does so at the expense of a deeper and more complex understanding of the possibilities of creative experimentation in the artist’s studio. By maintaining an hermetic approach to medium, it does not prepare students sufficiently for the reality of art making in the twenty-first century. In fact, by pretending that there is a select range of techniques fundamental to the artist’s trade, discipline-based teaching can often appear to be more engaged with the notion of skills preservation than purposeful art training. If art schools are to survive and prosper in an increasingly vocationally-oriented university environment, they need to fully synthesise the professional reality of contemporary art practice into their approach to teaching and learning. This paper discusses the way in which the ‘open’ studio approach to visual art study at QUT endeavours to incorporate the diversity and complexity of contemporary art while preserving the sense of collective purpose that discipline-based teaching fosters. By allowing students to independently develop their own art practices while also applying collaborative models of learning and assessment, the QUT studio program aims to equip students with a strong sense of self-reliance, a broad awareness and appreciation of contemporary art, and a deep understanding of studio-based experimentation unfettered by the boundaries of traditional media: all skills fundamental to the practice of contemporary art.

Performance of public private partnerships : an evolutionary perspective

PPPs are held to be a powerful way of mobilising private finance and resources to deliver public infrastructure. Theoretically, research into procurement has begun to acknowledge difficulties with the classification and assessment of different types of procurement, particularly those which do not sufficiently acknowledge variety within specific types of procurement methods. This paper advances a theoretical framework based on an evolutionary economic conceptualisation of a routine, which can accommodate the variety evident in procurement projects, in particular PPPs. The paper tests how the various elements of a PPP, as advanced in the theoretical framework, affect performance across 10 case studies. It concludes, that a limited number of elements of a PPP affect their performance, and provides strong evidence for the theoretical model advanced in this paper.

Mining trading partners’ preferences for efficient multi-issue bargaining in E‑business

Classical negotiation models are weak in supporting real-world business negotiations because these models often assume that the preference information of each negotiator is made public. Although parametric learning methods have been proposed for acquiring the preference information of negotiation opponents, these methods suffer from the strong assumptions about the specific utility function and negotiation mechanism employed by the opponents. Consequently, it is difficult to apply these learning methods to the heterogeneous negotiation agents participating in e‑marketplaces. This paper illustrates the design, development, and evaluation of a nonparametric negotiation knowledge discovery method which is underpinned by the well-known Bayesian learning paradigm. According to our empirical testing, the novel knowledge discovery method can speed up the negotiation processes while maintaining negotiation effectiveness. To the best of our knowledge, this is the first nonparametric negotiation knowledge discovery method developed and evaluated in the context of multi-issue bargaining over e‑marketplaces.

Assessment by interview and portfolio in a Graduate School Program

Assessment plays an integral role in teaching and learning in Higher Education and teachers have a strong interest in debates and commentaries on assessment as and for learning. In a one-year graduate entry teacher preparation program, the temptation is to emphasize assessment in an attempt to ensure students “cover” everything as part of a robust preparation for the profession. The risk is that, for students, assessment drives curriculum, and time spent in the completion of assignments is no guarantee of either effective learning or authentic preparation for teaching. Interviews as assessment provide an opportunity for a learning experience as well as an authentic task, since students will shortly be interviewing for employment in a “real world” situation. This paper reports on a project experimenting with interview panels as authentic assessment with pre-service early childhood teachers. At the end of their first semester of study, students enrolled in the Graduate Diploma of Education program at the Queensland University of Technology in Australia were required to participate in a panel interview where they were graded by a panel made up of three faculty staff and one undergraduate student enrolled in the four-year Bachelor of Education program. Students and panel members completed a questionnaire on their experience after the interview. Results indicated that both students and staff valued the experience and felt it was authentic. Results are discussed in terms of how the assessment interview and portfolio presentation supports graduating students in their preparation for employment interviews, and how this authentic assessment task has benefits for both students and teaching staff.

Measuring quality : how does this improve construction performance?

The paper charts the history and development of the Hong Kong Housing Department (HKHD) Performance Assessment Scoring System (PASS) from 1990 to the present day and examines its effect on facilitating change to the quality of construction work of building contractors engaged in the production of public sector housing projects Hong Kong. The paper builds partly on empirical research carried out by the author as part of a doctoral thesis from 2000 to 2005, on experiential knowledge and also on some relevant case studies. The outcomes from this earlier research and validation of PASS based on results derived from the system since the research was originally undertaken are of benefit to practitioners and academics working and studying in the areas of performance assessment and organisational culture and change. The conclusions presented in the paper further underpin the connection established in previous research between strong organisational culture and project and corporate success. Organisational culture was measured using a survey instrument originally developed by Denison and Neale (1994), adapted for the environment of the study, and corporate success was measured by the PASS system mentioned above. The major results of the original study indicate that there is significant linkage between strong organisational cultures and business success and the detailed findings were that, (1) strong organisational culture was positively associated a high level of company effectiveness, (2) a high level of company effectiveness was positively associated with the cultural traits of ‘consistency’, ‘adaptability’ and ‘mission’, and (3) a high level of company effectiveness was positively associated with the combined cultural traits represented by the dimensions of ‘external focus’ and ‘stable culture’. Several opportunities to take forward this research have been identified, including extending the study to other countries and also longitudinally re-evaluating some of the original case studies to ascertain how organisational cultures have changed or further developed in relation to the changing construction climate in Hong Kong.

Imagery and strength of craving for eating, drinking and playing sport

The elaborated intrusion (EI) theory of desire (Kavanagh, Andrade, & May, 2005) attributes the motivational force of cravings to cognitive elaboration, including imagery, of apparently spontaneous thoughts that intrude into awareness. We report a questionnaire study in which respondents rated a craving for food or drink. Questionnaire items derived from EI theory formed a single factor alongside factors for anticipated reward/relief, resistance, and opportunity. In a multiple regression predicting strength of craving, the first three factors accounted for 36% of the variance. Opportunity did not enter the model. In a second study, the difference between individuals' strong and weak cravings to take part in a sporting activity was shown to be related to visual, auditory, and general imagery, and to anticipated reward or relief from engaging in the activity. Implications for treatment of craving-related disorders are discussed in the light of these results and of other research indicating that interference with imagery can reduce the strength of craving.