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.

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.

Development of a controlled drinking self-efficacy scale and appraising its relation to alcohol dependence

There is no specific self-efficacy measure that has been developed primarily for problem drinkers seeking a moderation drinking goal. In this article, we report the factor structure of a 20-item Controlled Drinking Self-Efficacy Scale (CDSES; Sitharthan et al., 1996; Sitharthan et al., 1997). The results indicate that the CDSES is highly reliable, and the factor analysis using the full sample identified four factors: negative affect, positive mood/social context, frequency of drinking, and consumption quantity. A similar factor structure was obtained for the subsample of men. In contrast, only three factors emerged in the analysis of data on female participants. Compared to women, men had low self-efficacy to control their drinking in situations relating to positive mood/social context, and subjects with high alcohol dependence had low self-efficacy for situations relating to negative affect, social situations, and drinking less frequently. The CDSES can be a useful measure in treatment programs providing a moderation drinking goal.

Can Risk Management boost the supply of Affordable Housing Development and Management?

A shortage of affordable housing is a major problem in Australia today. This is mainly due to the limited supply of affordable housing that is provided by the non-government housing sector. Some private housing developers see the provision of affordable housing for lower income people as a high risk investment which offers a lower return than broader market-based housing. The scarcity of suitable land, a limited government ‘subsidy’, and increasing housing costs have not provided sufficient development incentives to encourage their investment despite the existing high demand for affordable housing. This study analyses the risk management process conducted by some private and not-for-profit housing providers in South East Queensland, and draws conclusions about the relationship between risk assessments/responses and past experiences. In-depth interviews of selected non-government housing providers have been conducted to facilitate an understanding of their approach to risk assessment/response in developing and in managing affordable housing projects. These developers use an informal risk management process as part of their normal business process in accordance with industry standards. A simple qualitative matrix has been used to analyse probability and impacts using a qualitative scale - low, medium and high. For housing providers who have considered investing in affordable housing but have not yet implemented any such projects, affordable housing development is seen as an opportunity that needs to be approached with caution. The risks associated with such projects and the levels of acceptance of these are not consistently identified by current housing providers. Many interviewees agree that the recognition of financial risk and the fear of community rejection of such housing projects have restrained them from committing to such investment projects. This study suggests that implementing improvements to the risk mitigation and management framework may assist in promoting the supply of affordable housing by non-government providers.

Investigation into Travel of Transit Oriented Development Residents’ of Kelvin Grove Urban Village

Travel surveys were conducted for collecting data related to residents’ travel at Kelvin Grove Urban Village (KGUV). Currently, KGUV has residents living in the affordable apartments, apartments, townhouses and student accommodation. As a part of data collection process, travel surveys were undertaken for residents living in apartments, affordable apartments and student accommodation. This document contains the questionnaire form used to collect the demographic and travel data related to residents at KGUV. A mail back survey technique was used to collect data for residents living in affordable apartment and apartments, and an intercept surveys was conducted for residents living in student accommodation.

Persistent chlamydial infections : role in inflammation and challenges for vaccine development.

The development of chlamydial vaccines continues to be a major challenge for researchers. While acute infections are the main target of vaccine development groups, Chlamydia is well known for its ability to establish chronic or persistent infections in its host. To date, little effort has focussed specifically on the challenges of vaccines to successfully combat the chronic or persistent phase of the disease and yet this will be a necessary aspect of any fully successful chlamydial vaccine. This short review specifically examines the phenomenon of chlamydial persistence and the unique challenges that this brings to vaccine development.

Waterfront Development for Residential Property in Malaysia

Rivers and water are valuable natural resources for human life, environment and national development. Recognition of water resources as national heritage will contribute towards more long term sustainable property development. Waterfront development is already a well-established phenomenon internationally. In Malaysia, as the economy began to change in 1980s, so did the land uses along many of the river and waterfront locations. The pressures of new technology coupled with an urban population growth and urbanization began to force a transition from water dependent industry to a variety of non-water dependent developments such as apartments, offices, and retail shopping areas. Residential waterfront development has taken advantage of available land and water amenities and incorporated as a feature or “selling point” of the development. It has been found that wide views of water add an average of 59% to the value of waterfront property, as well as providing attractive landscaping and better property neighborhoods respectively. Development of waterfront lands in Malaysia occurred with limited federal, state, or municipal planning guidance; resulting in cost aspects like flooding and pollution. Although some waterfront development projects continue to remain profitable with a maintained successful public access component, many have not. This paper provides a brief introduction to the research project to address this issue, which is currently on-going.

The explicit hazard model – part 1: theoretical development

Modern Engineering Asset Management (EAM) requires the accurate assessment of current and the prediction of future asset health condition. Appropriate mathematical models that are capable of estimating times to failures and the probability of failures in the future are essential in EAM. In most real-life situations, the lifetime of an engineering asset is influenced and/or indicated by different factors that are termed as covariates. Hazard prediction with covariates is an elemental notion in the reliability theory to estimate the tendency of an engineering asset failing instantaneously beyond the current time assumed that it has already survived up to the current time. A number of statistical covariate-based hazard models have been developed. However, none of them has explicitly incorporated both external and internal covariates into one model. This paper introduces a novel covariate-based hazard model to address this concern. This model is named as Explicit Hazard Model (EHM). Both the semi-parametric and non-parametric forms of this model are presented in the paper. The major purpose of this paper is to illustrate the theoretical development of EHM. Due to page limitation, a case study with the reliability field data is presented in the applications part of this study.

Problem-based learning (PBL) in Virtual Space : Developing experiences for professional development

This book reports the outcomes of an investigation into discovering the qualitatively different ways that students experience Problem-based learning (PBL)in virtual space. PBL is increasingly being used in many fields including engineering education. At the same time, many engineering education providers are turning to online distance education. Unfortunately there is a dearth of research into what constitutes an effective learning experience for adult learners who undertake PBL instruction through online distance education. Data were collected from a course which adopted the PBL strategy and was delivered entirely in virtual space. Students were asked to respond to open-ended questions designed to elicit their learning experiences. Data were analysed using the phenomenographic approach. Five qualitatively different ways of experiencing PBL in virtual space were discovered. Results indicate that the design of students' online learning experience was responsible for making students aware of deeper ways of experienceing PBL in virtual space. The outcomes imply that pedagogical strategies can be devised for shifting students' focus as they engage in virtual PBL.

Contemporary behavioural influences in an organisational setting and implications for intervention development

Professional drivers and safety Within the industrialised world, work-related crashes are the most common cause of work-related death, injury and reduced productivity (Charbotel et al., 2001; Toscano and Windau, 1994). Likewise in Australia, road crashes are the most common cause of work-related fatalities, injuries and absence from work (Haworth et al., 2000), with the average time lost being greater than any other workplace claim (Stewart-Bogle, 1999). There are obvious costs related to work crashes such as vehicle and property repair costs. There are also many hidden expenses including third party costs, workers compensation, medical costs, rehabilitation, customer-related costs, increased insurance premiums, administrative costs, legal fees and loss of productivity (Collingwood, 1997; Haworth et al, 2000).

Cues to starting CPAP in obstructive sleep apnoea : development and validation of the cues to CPAP Use Questionnaire (CCUQ)

Background: The reasons that a patient has to start treatment, their “Cues to Action”, are important for determining subsequent health behaviours. Cues to action are an explicit component of the Health Belief Model of CPAP acceptance adherence. At present there is no scale available to measure this construct for individuals with Obstructive Sleep Apnoea (OSA). This paper aims to develop, validate and describe responding patterns within an OSA patient sample to the Cues to CPAP Use Questionnaire (CCUQ).----- Method: Participants were 63 adult patients diagnosed with OSA who had never tried CPAP when initially recruited. The CCUQ was completed at one month after being prescribed CPAP.----- Results: Exploratory factor analysis (EFA) showed a three factor structure of the 9-item CCUQ, with “Health Cues”, “Partner Cues” and “Health Professional Cues” subscales accounting for 59.91% of the total variance. The CCUQ demonstrated modest internal consistency and split-half reliability. The questionnaire is brief and user-friendly, with readability at a 7th grade level. The most frequently endorsed cues for starting CPAP were Health Professional Cues (prompting by the sleep physician) and Health Cues such as tiredness and concern about health outcomes.----- Conclusions: This study validates a measure of an important motivational component of the Health Belief Model. Health Professional Cues and internal Health Cues were reported to be the most important prompts to commence CPAP by this patient sample.

Age-related development of a refractive index plateau in the human lens : evidence for a distinct nucleus

The human lens comprises two distinct regions in which the refractive index changes at different rates. The periphery contains a rapidly increasing refractive index gradient, which becomes steeper with age. The inner region contains a shallow gradient, which flattens with age, due to formation of a central plateau, of RI = 1.418, which reaches a maximum size of 7.0 × 3.05 mm around age 60 years. Formation of the plateau can be attributed to compression of fibre cells generated in prenatal life. Present in prenatal but not in postnatal fibre cells, γ-crystallin may play a role in limiting nuclear cell compression.

Development and validation of 3-Minute Nutrition Screening (3-MinNS) tool for acute hospital patients in Singapore

It is important to detect and treat malnutrition in hospital patients so as to improve clinical outcome and reduce hospital stay. The aim of this study was to develop and validate a nutrition screening tool with a simple and quick scoring system for acute hospital patients in Singapore. In this study, 818 newly admitted patients aged above 18 years old were screened using five parameters that contribute to the risk of malnutrition. A dietitian blinded to the nutrition screening score assessed the same patients using the reference standard, Subjective Global Assessment (SGA) within 48 hours. The sensitivity and specificity were established using the Receiver Operator Characteristics (ROC) curve and the best cutoff scores determined. The nutrition parameter with the largest Area Under the ROC Curve (AUC) was chosen as the final screening tool, which was named 3-Minute Nutrition Screening (3-MinNS). The combination of the parameters weight loss, intake and muscle wastage (3-MinNS), gave the largest AUC when compared with SGA. Using 3-MinNS, the best cutoff point to identify malnourished patients is three (sensitivity 86%, specificity 83%). The cutoff score to identify subjects at risk of severe malnutrition is five (sensitivity 93%, specificity 86%). 3-Minute Nutrition Screening is a valid, simple and rapid tool to identify patients at risk of malnutrition in Singapore acute hospital patients. It is able to differentiate patients at risk of moderate malnutrition and severe malnutrition for prioritization and management purposes.