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.

Ipswich stories : using digital storytelling to research intergenerational sharing of heritage, history, place, identity and community connection : a pilot study

Co-creative media production practices offer important new modes and opportunities for social participation and engagement. In mid-2009 Institute for Creative Industries and Innovation researchers at QUT adapted a specific model of co-creative media production, known as ‘digital storytelling’ and piloted it as an action research platform for facilitating and researching knowledge production based on intergenerational dialogue and exchange. Nine stories were produced and important insights were generated into this particular use of digital storytelling, as well as the impact of institutional constraints and opportunities on the possibilities and outcomes co-creative media practices and processes.

Conducting research in herbal and complementary medicine

This paper outlines information and advice on how a practitioner can formally pursue research pertaining to herbal or complementary medicine. It recommends five practical steps: get advice and acquire skills, find out what other people have done already, consider what research you want to do, decide on a design and finalise a detailed research plan. Enrolling in a postgraduate research degree program is recommended as a way to acquire basic research skills and obtain support for an initial project.

Action research for contexts of change and inequality

Undoubtedly, the past half-century has witnessed an escalation of changes in the social, political, economic and educational structures in many societies around the world. Some have seen change as a challenge and hope while, for many others, it is a source of concern and worry. Some have adopted change with gusto, while for many it is something to be resisted. Some say we live in a world and times with an increasing awareness that “times are changing”, while for some “the more things change, the more they stay the same”.

Beating the urge : implications of research into substance-related desires

Despite the advent of improved pharmacological treatments to alleviate substance-related desires, psychological approaches will continue to be required. However, the current psychological treatment that most specifically focuses on desires and their management—cue exposure (CE)—has not lived up to its original promise. This paper argues that current psychological approaches to desire do not adequately incorporate our knowledge about the factors that trigger, maintain, and terminate episodes of desire. It asserts that the instigation and maintenance of desires involve both associative and elaborative processes. Understanding the processes triggering the initiation of intrusive thoughts may assist in preventing some episodes, but occasional intrusions will be inevitable. A demonstration of the ineffectiveness of thought suppression may discourage its use as a coping strategy for desire-related intrusions, and mindfulness meditation plus cognitive therapy may help in accepting their occurrence and letting them go. Competing tasks may be used to reduce elaboration of desires, and competing sensory images may have particular utility. The application of these procedures during episodes that are elicited in the clinic may allow the acquisition of more effective strategies to address desires in the natural environment.

Choreographing research : supervising the dancing thesis

The dancing doctorate is an interrogative endeavour which can but nurture the art form and forge a beneficial dynamism between those who seek and those who assess the emerging knowledges of dance’. (Vincs, 2009) From 2006-2008 three dance academics from Perth, Brisbane and Melbourne undertook a research project entitled Dancing between Diversity and Consistency: Refining Assessment in Postgraduate Degrees in Dance, funded by the ALTC Priority Projects Program. Although assessment rather than supervision was the primary focus of this research, interviews with 40 examiner/supervisors, 7 research deans and 32 candidates across Australia and across the creative arts, primarily in dance, provide an insight into what might be considered best practice in preparing students for higher research degrees, and the challenges that embodied and experiential knowledges present for supervision. The study also gained the industry perspectives of dance professionals in a series of national forums in 5 cities, based around the value of higher degrees in dance. The qualitative data gathered from these two primary sources was coded and analysed using the NVivo system. Further perspectives were drawn from international consultant and dance researcher Susan Melrose, as well as recent publications in the field. Dance is a young addition to academia and consequently there tends to be a close liaison between the academy and the industry, with a relational fluidity that is both beneficial and problematic. This partially explains why dance research higher degrees are predominantly practice-led (or multi-modal, referring to those theses where practice comprises the substantial examinable component). As a physical, embodied art form, dance engages with the contested territory of legitimising alternative forms of knowledge that do not sit comfortably with accepted norms of research. In supporting research students engaged with dance practice, supervisors traverse the tricky terrain of balancing university academic requirements with studies that are emergent, not only in the practice and attendant theory but in their methodologies and open-ended outcomes; and in an art form in which originality and new knowledge also arises from collaborative creative processes. Formal supervisor accreditation through training is now mandatory in most Australian universities, but it tends to be generic and not address supervisory specificity. This paper offers the kind of alternative proposed by Edwards (2002) that improving postgraduate supervision will be effective if supervisors are empowered to generate their own standards and share best practice; in this case, in ways appropriate to the needs of their discipline and alternative modes of thesis presentation. In order to frame the qualities and processes conducive to this goal, this paper will draw on both the experiences of interviewees and on philosophical premises which underpin the research findings of our study. These include the ongoing challenge of dissolving the binary oppositions of theory and practice, especially in creative arts practice where theory resides in and emerges from the doing as much as in articulating reflection about the doing through what Melrose (2003) terms ‘mixed mode disciplinary practices’. In guiding practitioners through research higher degrees, how do supervisors deal with not only different forms of knowledge but indeed differing modes of knowledge? How can they navigate tensions that occur between the ‘incompatible competencies’ (Candlin, 2000) of the ‘spectating’ academic experts with their ‘irrepressible drive ... to inscribe, interpret, and hence to practise temporal closure’, and practitioner experts who create emergent works of ‘residual unfinishedness’ (Melrose 2006) which are not only embodied but ephemeral, as in the case of live performance?

New routes for the field trip

As with the broader field of education research, most writing on the subject of school excursions and field trips has centred around progressive/humanist concerns for building pupil’s self-esteem and for the development of the ‘whole child’. Such research has also stressed the importance of a broad, grounded, and experiential curriculum - as exemplified by subjects containing these extra-school activities - as well as the possibility of strengthening the relationship between student and teacher. Arguing that this approach to the field trip is both exhausted of ideas and conceptually flawed, this paper proposes some alternate routes into the area for the prospective researcher. First, it is argued that by historicising the subject matter, it can be seen that school excursions are not simply the product of the contemporary humanist desire for diverse and fulfilling educational experiences, rather they can, in part, be traced to eighteenth century beliefs among the English gentry that travel formed a crucial component of a good education, to the advent of an affordable public rail system, and to school tours associated with the Temperance movement. Second, field trips can be understood from within the associated framework of concerns over the governance of tourism and the organisation of disciplinary apparatuses for the production of an educated and regulated citizenry. Far from being a simple learning experience, museums and art galleries form part of a complex of disciplinary and power relations designed to produce a populace with very specific capacities, aspirations and styles of public conduct. Finally, rather than allowing children ‘freedom’ from the constraints of the classroom, on the contrary, through the medium of the field-trip, children can become accustomed to having their activities governed in the broader domain of the generalised community . School excursions thereby constitute an effective tactic through which young people have their conduct managed, and their social and scholastic identities shaped and administered.

A news story as big as a doctoral thesis? : deploying journalistic methodology in academic research

The purpose of this paper is to identify and recommend the emergence of an academic research methodology for Journalism the academic discipline, through reviewing various journalistic methods of research – those making up a key element in such methodology. Its focus is on journalistic styles of work employed in academic contexts especially research on mass media issues. It proposes that channelling such activity into disciplined academic forms will enhance both: allowing the former to provide more durable and deeper outcomes, injecting additional energy and intensity of purpose into the latter. It will briefly consider characteristics of research methodologies and methods, generally; characteristics of the Journalism discipline, and its relationship with mass media industries and professions. The model of journalism used here is the Western liberal stream. A proposition is made, that teaching and research in universities focused on professional preparation of journalists, has developed so that it is a mature academic discipline. Its adherents are for the most part academics with background in journalistic practice, and so able to deploy intellectual skills of journalists, while also accredited with Higher Degrees principally in humanities. Research produced in this discipline area stands to show two characteristics: (a) it employs practices used generally in academic research, e.g. qualitative research methods such as ethnographic studies or participant observation, or review of documents including archived media products, and (b) within such contexts it may use more specifically journalistic techniques, e.g. interviewing styles, reflection on practice of journalism, and in creative practice research, journalistic forms of writing – highlighting journalistic / practitioner capabilities of the author. So the Journalism discipline, as a discipline closely allied to a working profession, is described as one where individual professional skills and background preparation for media work will be applicable to academic research. In this connection the core modus operandi will be the directly research-related practices of: insistent establishment of facts, adept crafting of reportage, and economising well with time. Prospective fields for continuing research are described:- work in new media; closer investigation of relations among media producers and audiences; journalism as creative practice, and general publishing by journalists, e.g. writing histories.

Education research and youth subculture theory

Recent theoretical developments (particularly those involving the work of Michel Foucault) have challenged the foundations upon which subcultural analysis is built. It is not the intention of this paper to take issue with the underpinning neo-Marxist assumptions concerning hegemony, consciousness and culture. Rather, the intention is simply to problematise the use of subculture theory in terms of its totalising, normalising and dichotomising nature.

Virtual reality or virtually real : blended teaching and learning in a Master's level research methods class

This paper examines the enabling effect of using blended learning and synchronous internet mediated communication technologies to improve learning and develop a Sense of Community (SOC) in a group of post-graduate students consisting of a mix of on-campus and off-campus students. Both quantitative and qualitative data collected over a number of years supports the assertion that the blended learning environment enhanced both teaching and learning. The development of a SOC was pivotal to the success of the blended approach when working with geographically isolated groups within a single learning environment.

Can theories about leadership in schools inform the work of Principals in schools?

This paper explores recent theorising on the ways in which Principals exercise leadership in their schools with reference to the Leading 21st Century Schools Project in Australia. First, it provides an historical overview of approaches to leadership. Second, it utilises a rhetorical question about leadership to analyse the ways in which leadership and management tensions pose challenges to Principals' efforts to capacity build their staff. Third, it and suggests that the notion of distributed leadership has been the most useful method in fostering Asia literacy in the Leading 21st Century Schools Project.

Writing together, learning together : the value and effectiveness of a research writing group for doctoral students

The high level of scholarly writing required for a doctoral thesis is a challenge for many research students. However, formal academic writing training is not a core component of many doctoral programs. Informal writing groups for doctoral students may be one method of contributing to the improvement of scholarly writing. In this paper, we report on a writing group that was initiated by an experienced writer and higher degree research supervisor to support and improve her doctoral students’ writing capabilities. Over time, this group developed a workable model to suit their varying needs and circumstances. The model comprised group sessions, an email group, and individual writing. Here, we use a narrative approach to explore the effectiveness and value of our research writing group model in improving scholarly writing. The data consisted of doctoral students’ reflections to stimulus questions about their writing progress and experiences. The stimulus questions sought to probe individual concerns about their own writing, what they had learned in the research writing group, the benefits of the group, and the disadvantages and challenges to participation. These reflections were analysed using thematic analysis. Following this analysis, the supervisor provided her perspective on the key themes that emerged. Results revealed that, through the writing group, members learned technical elements (e.g., paragraph structure), non-technical elements (e.g., working within limited timeframes), conceptual elements (e.g., constructing a cohesive arguments), collaborative writing processes, and how to edit and respond to feedback. In addition to improved writing quality, other benefits were opportunities for shared writing experiences, peer support, and increased confidence and motivation. The writing group provides a unique social learning environment with opportunities for: professional dialogue about writing, peer learning and review, and developing a supportive peer network. Thus our research writing group has proved an effective avenue for building doctoral students’ capability in scholarly writing. The proposed model for a research writing group could be applicable to any context, regardless of the type and location of the university, university faculty, doctoral program structure, or number of postgraduate students. It could also be used within a group of students with diverse research abilities, needs, topics and methodologies. However, it requires a group facilitator with sufficient expertise in scholarly writing and experience in doctoral supervision who can both engage the group in planned writing activities and also capitalise on fruitful lines of discussion related to students’ concerns as they arise. The research writing group is not intended to replace traditional supervision processes nor existing training. However it has clear benefits for improving scholarly writing in doctoral research programs particularly in an era of rapidly increasing student load.

Strategic short termism as an issue of top-teams’ temporal orientation

Short-termism among firms, the tendency to excessively discount long-term benefits and favour less valuable short-term benefits, has been a prominent issue in business and public policy debates but research to date has been inconclusive. We study how managers frame, interpret, and resolve problems of intertemporal choice in actual decisions by using computer aided text analysis to measure the frequency of top-team temporal references in 1653 listed Australian firms between 1992-2005. Contrary to short-termism arguments we find evidence of a significant general increase in Future orientation and a significant decrease in Current/Past orientation. We also show top-teams’ temporal orientation is related to their strategic orientation, specifically the extent to which they focus on Innovation-Expansion and Capacity Building.

Sampling perceptions; testing reality : an evidence-based approach to measurably improve information literacy and student research skills

Do commencing students possess the level of information literacy (IL) knowledge and skills they need to succeed at university? What impact does embedding IL within the engineering and design curriculum have? This paper reports on the self-perception versus the reality of IL knowledge and skills, across a large cohort of first year built environment and engineering students. Acting on the findings of this evaluation, the authors (a team of academic librarians) developed an intensive IL skills program which was integrated into a faculty wide unit. Perceptions, knowledge and skills were re-evaluated at the end of the semester to determine if embedded IL education made a difference. Findings reveal that both the perception and reality of IL skills were significantly and measurably improved.