The ghrelin receptor isoforms (GHS-R1a and GHS-R1b) and GPR39 : an investigation into receptor dimerisation

Prostate cancer is the second most common cause of cancer-related deaths in Western males. Current diagnostic, prognostic and treatment approaches are not ideal and advanced metastatic prostate cancer is incurable. There is an urgent need for improved adjunctive therapies and markers for this disease. GPCRs are likely to play a significant role in the initiation and progression of prostate cancer. Over the last decade, it has emerged that G protein coupled receptors (GPCRs) are likely to function as homodimers and heterodimers. Heterodimerisation between GPCRs can result in the formation of novel pharmacological receptors with altered functional outcomes, and a number of GPCR heterodimers have been implicated in the pathogenesis of human disease. Importantly, novel GPCR heterodimers represent potential new targets for the development of more specific therapeutic drugs. Ghrelin is a 28 amino acid peptide hormone which has a unique n-octanoic acid post-translational modification. Ghrelin has a number of important physiological roles, including roles in appetite regulation and the stimulation of growth hormone release. The ghrelin receptor is the growth hormone secretagogue receptor type 1a, GHS-R1a, a seven transmembrane domain GPCR, and GHS-R1b is a C-terminally truncated isoform of the ghrelin receptor, consisting of five transmembrane domains. Growing evidence suggests that ghrelin and the ghrelin receptor isoforms, GHS-R1a and GHS-R1b, may have a role in the progression of a number of cancers, including prostate cancer. Previous studies by our research group have shown that the truncated ghrelin receptor isoform, GHS-R1b, is not expressed in normal prostate, however, it is expressed in prostate cancer. The altered expression of this truncated isoform may reflect a difference between a normal and cancerous state. A number of mutant GPCRs have been shown to regulate the function of their corresponding wild-type receptors. Therefore, we investigated the potential role of interactions between GHS-R1a and GHS-R1b, which are co-expressed in prostate cancer and aimed to investigate the function of this potentially new pharmacological receptor. In 2005, obestatin, a 23 amino acid C-terminally amidated peptide derived from preproghrelin was identified and was described as opposing the stimulating effects of ghrelin on appetite and food intake. GPR39, an orphan GPCR which is closely related to the ghrelin receptor, was identified as the endogenous receptor for obestatin. Recently, however, the ability of obestatin to oppose the effects of ghrelin on appetite and food intake has been questioned, and furthermore, it appears that GPR39 may in fact not be the obestatin receptor. The role of GPR39 in the prostate is of interest, however, as it is a zinc receptor. Zinc has a unique role in the biology of the prostate, where it is normally accumulated at high levels, and zinc accumulation is altered in the development of prostate malignancy. Ghrelin and zinc have important roles in prostate cancer and dimerisation of their receptors may have novel roles in malignant prostate cells. The aim of the current study, therefore, was to demonstrate the formation of GHS-R1a/GHS-R1b and GHS-R1a/GPR39 heterodimers and to investigate potential functions of these heterodimers in prostate cancer cell lines. To demonstrate dimerisation we first employed a classical co-immunoprecipitation technique. Using cells co-overexpressing FLAG- and Myc- tagged GHS-R1a, GHS-R1b and GPR39, we were able to co-immunoprecipitate these receptors. Significantly, however, the receptors formed high molecular weight aggregates. A number of questions have been raised over the propensity of GPCRs to aggregate during co-immunoprecipitation as a result of their hydrophobic nature and this may be misinterpreted as receptor dimerisation. As we observed significant receptor aggregation in this study, we used additional methods to confirm the specificity of these putative GPCR interactions. We used two different resonance energy transfer (RET) methods; bioluminescence resonance energy transfer (BRET) and fluorescence resonance energy transfer (FRET), to investigate interactions between the ghrelin receptor isoforms and GPR39. RET is the transfer of energy from a donor fluorophore to an acceptor fluorophore when they are in close proximity, and RET methods are, therefore, applicable to the observation of specific protein-protein interactions. Extensive studies using the second generation bioluminescence resonance energy transfer (BRET2) technology were performed, however, a number of technical limitations were observed. The substrate used during BRET2 studies, coelenterazine 400a, has a low quantum yield and rapid signal decay. This study highlighted the requirement for the expression of donor and acceptor tagged receptors at high levels so that a BRET ratio can be determined. After performing a number of BRET2 experimental controls, our BRET2 data did not fit the predicted results for a specific interaction between these receptors. The interactions that we observed may in fact represent ‘bystander BRET’ resulting from high levels of expression, forcing the donor and acceptor into close proximity. Our FRET studies employed two different FRET techniques, acceptor photobleaching FRET and sensitised emission FRET measured by flow cytometry. We were unable to observe any significant FRET, or FRET values that were likely to result from specific receptor dimerisation between GHS-R1a, GHS-R1b and GPR39. While we were unable to conclusively demonstrate direct dimerisation between GHS-R1a, GHS-R1b and GPR39 using several methods, our findings do not exclude the possibility that these receptors interact. We aimed to investigate if co-expression of combinations of these receptors had functional effects in prostate cancers cells. It has previously been demonstrated that ghrelin stimulates cell proliferation in prostate cancer cell lines, through ERK1/2 activation, and GPR39 can stimulate ERK1/2 signalling in response to zinc treatments. Additionally, both GHS-R1a and GPR39 display a high level of constitutive signalling and these constitutively active receptors can attenuate apoptosis when overexpressed individually in some cell types. We, therefore, investigated ERK1/2 and AKT signalling and cell survival in prostate cancer the potential modulation of these functions by dimerisation between GHS-R1a, GHS-R1b and GPR39. Expression of these receptors in the PC-3 prostate cancer cell line, either alone or in combination, did not alter constitutive ERK1/2 or AKT signalling, basal apoptosis or tunicamycin-stimulated apoptosis, compared to controls. In summary, the potential interactions between the ghrelin receptor isoforms, GHS-R1a and GHS-R1b, and the related zinc receptor, GPR39, and the potential for functional outcomes in prostate cancer were investigated using a number of independent methods. We did not definitively demonstrate the formation of these dimers using a number of state of the art methods to directly demonstrate receptor-receptor interactions. We investigated a number of potential functions of GPR39 and GHS-R1a in the prostate and did not observe altered function in response to co-expression of these receptors. The technical questions raised by this study highlight the requirement for the application of extensive controls when using current methods for the demonstration of GPCR dimerisation. Similar findings in this field reflect the current controversy surrounding the investigation of GPCR dimerisation. Although GHS-R1a/GHS-R1b or GHS-R1a/GPR39 heterodimerisation was not clearly demonstrated, this study provides a basis for future investigations of these receptors in prostate cancer. Additionally, the results presented in this study and growing evidence in the literature highlight the requirement for an extensive understanding of the experimental method and the performance of a range of controls to avoid the spurious interpretation of data gained from artificial expression systems. The future development of more robust techniques for investigating GPCR dimerisation is clearly required and will enable us to elucidate whether GHS-R1a, GHS-R1b and GPR39 form physiologically relevant dimers.

Enhancing the transition of commencing students into university : an institution-wide approach

The importance of the first year experience (FYE) to success at university is well documented and supported with the transition into university regarded as crucial. While there is also support for the notion that a successful FYE should have a whole-of-institution focus and models have been proposed, many institutions still face challenges in achieving institution-wide FYE program implementation. This paper discusses the origins, theoretical and empirical bases and structure of an institution-wide approach to the FYE. It uses a case study of the Transitions In Project (TIP) at the Queensland University of Technology to illustrate how institution-wide FYE program implementation can be achieved and sustained. TIP had four inter-related projects focussing on at-risk students, first year curriculum, learning resources and staff development. The key aim of TIP was to identify good practice and institutionalise it in a sustainable way. The degree of success in achieving this is evaluated.

The CAPA Indigenous Project. Indigenous postgraduate education: A project into the barriers which indigenous students must overcome in postgraduate studies: A short history 1993-2000.

This brief paper outlines some of the early work undertaken in Indigenous postgraduate activism in Australia. In particular, the work undertaken in the lead up to the 'Project into the Barriers which Indigenous Students must Overcome in Postgraduate Studies'.

Enhancing pathways for ACE learners : Building the capacity of Adult and Community Education (ACE) providers to offer non-accredited courses and VET modules with pathways into VET programs.

This report presents findings from a project that considered a) the current capacity of Adult and Community Education (ACE) providers to offer non-accredited courses and single modules of accredited learning that provide pathways into full scale accredited VET programs, and b) the factors that aid and inhibit this from occurring. Based on the findings, suggestions are made as to what needs to be done to extend this capacity and thereby to achieve the goals outlined in the 2008 Ministerial Declaration on Adult Community Education.

Deterring drug drivers : a study into the initial impact of oral random roadside drug testing in Queensland

Objective: The global implementation of oral random roadside drug testing is relatively limited, and correspondingly, the literature that focuses on the effectiveness of this intervention is scant. This study aims to provide a preliminary indication of the impact of roadside drug testing in Queensland. Methods: A sample of Queensland motorists’ (N= 922) completed a self-report questionnaire to investigate their drug driving behaviour, as well as examine the perceived affect of legal sanctions (certainty, severity and swiftness) and knowledge of the countermeasure on their subsequent offending behaviour. Results: Analysis of the collected data revealed that approximately 20% of participants reported drug driving at least once in the last six months. Overall, there was considerable variability in respondent’s perceptions regarding the certainty, severity and swiftness of legal sanctions associated with the testing regime and a considerable proportion remained unaware of testing practices. In regards to predicting those who intended to drug driving again in the future, perceptions of apprehension certainty, more specifically low certainty of apprehension, were significantly associated with self-reported intentions to offend. Additionally, self-reported recent drug driving activity and frequent drug consumption were also identified as significant predictors, which indicates that in the current context, past behaviour is a prominent predictor of future behaviour. To a lesser extent, awareness of testing practices was a significant predictor of intending not to drug drive in the future. Conclusion: The results indicate that drug driving is relatively prevalent on Queensland roads, and a number of factors may influence such behaviour. Additionally, while the roadside testing initiative is beginning to have a deterrent impact, its success will likely be linked with targeted intelligence-led implementation in order to increase apprehension levels as well as the general deterrent effect.

It really is black and white when you look at it like this : reporting on a study into Australian professors' research and teaching priorities

Despite optimistic claims about the research-teaching nexus, Australian academics still face tension between research and teaching. The teaching and research priorities, beliefs and behaviours of 70 Professorial and Associate Professorial academics in Science, Information Technology and Engineering were examined in this study. The academics from 4 faculties in 3 Australian universities, were asked to rank 16 research activities and 16 matched learning and teaching (L&T) activities from each of three perspectives: job satisfaction, leadership behaviour, and perceptions of professional importance. The findings, which were remarkably consistent across the three universities, were unequivocally in favour of Research. The only L&T activity that was ranked consistently well was “Improving student satisfaction ratings for Teaching”. The data demonstrates that Australian government and university initiatives to raise the status of L&T activity are not impacting significantly on Australia’s future leaders of university learning.

Free surface flow into a horizontal slot with zero gravity

The two-dimensional free surface flow of a finite-depth fluid into a horizontal slot is considered. For this study, the effects of viscosity and gravity are ignored. A generalised Schwarz-Christoffel mapping is used to formulate the problem in terms of a linear integral equation, which is solved exactly with the use of a Fourier transform. The resulting free surface profile is given explicitly in closed-form.

New insights into Crustal Contributions to Large-volume Rhyolite Generation in the Mid-Tertiary Sierra Madre Occidental Province, Mexico, revealed by U Pb Geochronology

Voluminous (≥3·9 × 105 km3), prolonged (∼18 Myr) explosive silicic volcanism makes the mid-Tertiary Sierra Madre Occidental province of Mexico one of the largest intact silicic volcanic provinces known. Previous models have proposed an assimilation–fractional crystallization origin for the rhyolites involving closed-system fractional crystallization from crustally contaminated andesitic parental magmas, with <20% crustal contributions. The lack of isotopic variation among the lower crustal xenoliths inferred to represent the crustal contaminants and coeval Sierra Madre Occidental rhyolite and basaltic andesite to andesite volcanic rocks has constrained interpretations for larger crustal contributions. Here, we use zircon age populations as probes to assess crustal involvement in Sierra Madre Occidental silicic magmatism. Laser ablation-inductively coupled plasma-mass spectrometry analyses of zircons from rhyolitic ignimbrites from the northeastern and southwestern sectors of the province yield U–Pb ages that show significant age discrepancies of 1–4 Myr compared with previously determined K/Ar and 40Ar/39Ar ages from the same ignimbrites; the age differences are greater than the errors attributable to analytical uncertainty. Zircon xenocrysts with new overgrowths in the Late Eocene to earliest Oligocene rhyolite ignimbrites from the northeastern sector provide direct evidence for some involvement of Proterozoic crustal materials, and, potentially more importantly, the derivation of zircon from Mesozoic and Eocene age, isotopically primitive, subduction-related igneous basement. The youngest rhyolitic ignimbrites from the southwestern sector show even stronger evidence for inheritance in the age spectra, but lack old inherited zircon (i.e. Eocene or older). Instead, these Early Miocene ignimbrites are dominated by antecrystic zircons, representing >33 to ∼100% of the dated population; most antecrysts range in age between ∼20 and 32 Ma. A sub-population of the antecrystic zircons is chemically distinct in terms of their high U (>1000 ppm to 1·3 wt %) and heavy REE contents; these are not present in the Oligocene ignimbrites in the northeastern sector of the Sierra Madre Occidental. The combination of antecryst zircon U–Pb ages and chemistry suggests that much of the zircon in the youngest rhyolites was derived by remelting of partially molten to solidified igneous rocks formed during preceding phases of Sierra Madre Occidental volcanism. Strong Zr undersaturation, and estimations for very rapid dissolution rates of entrained zircons, preclude coeval mafic magmas being parental to the rhyolite magmas by a process of lower crustal assimilation followed by closed-system crystal fractionation as interpreted in previous studies of the Sierra Madre Occidental rhyolites. Mafic magmas were more probably important in providing a long-lived heat and material flux into the crust, resulting in the remelting and recycling of older crust and newly formed igneous materials related to Sierra Madre Occidental magmatism.

Assessment of commercial network RTK user positioning performance over long inter-station distances

The paper provides an assessment of the performance of commercial Real Time Kinematic (RTK) systems over longer than recommended inter-station distances. The experiments were set up to test and analyse solutions from the i-MAX, MAX and VRS systems being operated with three triangle shaped network cells, each having an average inter-station distance of 69km, 118km and 166km. The performance characteristics appraised included initialization success rate, initialization time, RTK position accuracy and availability, ambiguity resolution risk and RTK integrity risk in order to provide a wider perspective of the performance of the testing systems. ----- ----- The results showed that the performances of all network RTK solutions assessed were affected by the increase in the inter-station distances to similar degrees. The MAX solution achieved the highest initialization success rate of 96.6% on average, albeit with a longer initialisation time. Two VRS approaches achieved lower initialization success rate of 80% over the large triangle. In terms of RTK positioning accuracy after successful initialisation, the results indicated a good agreement between the actual error growth in both horizontal and vertical components and the accuracy specified in the RMS and part per million (ppm) values by the manufacturers. ----- ----- Additionally, the VRS approaches performed better than the MAX and i-MAX when being tested under the standard triangle network with a mean inter-station distance of 69km. However as the inter-station distance increases, the network RTK software may fail to generate VRS correction and then may turn to operate in the nearest single-base RTK (or RAW) mode. The position uncertainty reached beyond 2 meters occasionally, showing that the RTK rover software was using an incorrect ambiguity fixed solution to estimate the rover position rather than automatically dropping back to using an ambiguity float solution. Results identified that the risk of incorrectly resolving ambiguities reached 18%, 20%, 13% and 25% for i-MAX, MAX, Leica VRS and Trimble VRS respectively when operating over the large triangle network. Additionally, the Coordinate Quality indicator values given by the Leica GX1230 GG rover receiver tended to be over-optimistic and not functioning well with the identification of incorrectly fixed integer ambiguity solutions. In summary, this independent assessment has identified some problems and failures that can occur in all of the systems tested, especially when being pushed beyond the recommended limits. While such failures are expected, they can offer useful insights into where users should be wary and how manufacturers might improve their products. The results also demonstrate that integrity monitoring of RTK solutions is indeed necessary for precision applications, thus deserving serious attention from researchers and system providers.

Incorporating parameter uncertainty into quantitative microbial risk assessment (QMRA)

Modern statistical models and computational methods can now incorporate uncertainty of the parameters used in Quantitative Microbial Risk Assessments (QMRA). Many QMRAs use Monte Carlo methods, but work from fixed estimates for means, variances and other parameters. We illustrate the ease of estimating all parameters contemporaneously with the risk assessment, incorporating all the parameter uncertainty arising from the experiments from which these parameters are estimated. A Bayesian approach is adopted, using Markov Chain Monte Carlo Gibbs sampling (MCMC) via the freely available software, WinBUGS. The method and its ease of implementation are illustrated by a case study that involves incorporating three disparate datasets into an MCMC framework. The probabilities of infection when the uncertainty associated with parameter estimation is incorporated into a QMRA are shown to be considerably more variable over various dose ranges than the analogous probabilities obtained when constants from the literature are simply ‘plugged’ in as is done in most QMRAs. Neglecting these sources of uncertainty may lead to erroneous decisions for public health and risk management.

Expression and connection : the integration of the reflective learning process and the writing process into social network sites

A number of instructors have recently adopted social network sites (SNSs) for learning. However, the learning design of SNSs often remains at a preliminary level similar to a personal log book because it does not properly include reflective learning elements such as individual reflection and collaboration. This article looks at the reflective learning process and the public writing process as a way of improving the quality of reflective learning on SNSs. It proposes a reflective learning model on SNSs based on two key pedagogical concepts for social networking: individual expression and collaborative connection. It is expected that the model would be helpful for instructors in designing a reflective learning process on SNSs in an effective and flexible way.

Shifting the technology context : career-change entrants’ transition into teaching

Becoming a teacher in technology-rich classrooms is a complex and challenging transition for career-change entrants. Those with generic or specialist Information and Communication Technology (ICT) expertise bring a mindset about purposeful uses of ICT that enrich student learning and school communities. The transition process from a non-education environment is both enhanced and constrained by shifting the technology context of generic or specialist ICT expertise, developed through a former career as well as general life experience. In developing an understanding of the complexity of classrooms and creating a learner centred way of working, perceptions about learners and learning evolve and shift. Shifts in thinking about how ICT expertise supports learners and enhances learning preceded shifts in perceptions about being a teacher, working with colleagues, and functioning in schools that have varying degrees of intensity and impact on evolving professional identities. Current teacher education and school induction programs are seen to be falling short of meeting the needs of career-change entrants and, as a flow on, the students they nurture. Research (see, for example, Tigchelaar, Brouwer, & Korthagen, 2008; Williams & Forgasz, 2009) highlights the value of generic and specialist expertise career-change teachers bring to the profession and draws attention to the challenges such expertise begets (Anthony & Ord, 2008; Priyadharshini & Robinson-Pant, 2003). As such, the study described in this thesis investigated perceptions of career-change entrants, who have generic (Mishra & Koehler, 2006) or specialist expertise, that is, ICT qualifications and work experience in the use of ICT. The career-change entrants‘ perceptions were sought as they shifted the technology context and transitioned into teaching in technology-rich classrooms. The research involved an interpretive analysis of qualitative data and quantitative data. The study used the explanatory case study (Yin, 1994) methodology enriched through grounded theory processes (Strauss & Corbin, 1998), to develop a theory about professional identity transition from the perceptions of the participants in the study. The study provided insights into the expertise and experiences of career change entrants, particularly in relation to how professional identities that include generic and specialist ICT knowledge and expertise were reconfigured while transitioning into the teaching profession. This thesis presents the Professional Identity Transition Theory that encapsulates perceptions about teaching in technology-rich classrooms amongst a selection of the increasing number of career-change entrants. The theory, grounded in the data, (Strauss & Corbin, 1998) proposes that career-change entrants experience transition phases of varying intensity that impact on professional identity, retention and development as a teacher. These phases are linked to a shift in perceptions rather than time as a teacher. Generic and specialist expertise in the use of ICT is a weight of the past and an asset that makes the transition process more challenging for career-change entrants. The study showed that career-change entrants used their experiences and perceptions to develop a way of working in a school community. Their way of working initially had an adaptive orientation focussed on immediate needs as their teaching practice developed. Following a shift of thinking, more generative ways of working focussed on the future emerged to enable continual enhancement and development of practice. Sustaining such learning is a personal, school and systemic challenge for the teaching profession.

New research literacies for contemporary research into literacy and new media?

The term literacy remains highly contested and debates continue about how literacy might best be researched and to what ends. For some, literacy is simply a matter of acquiring the technical competence which enables people to read and write. Literacy research conducted from this point of view does not usually concern itself with the new media but rather focuses on how people learn to code and decode print text. For others, however, literacy is more complex and involves learning a repertoire of practices for communicating and getting things done in particular social and cultural contexts. Literacy research conducted from this sociocultural point of view accepts that the new media are central to the field because in everyday cultural practice people are using the new media to make meaning, to express themselves and to communicate and work with others. Socio-cultural approaches to literacy research have already provided rich material which has assisted educators to understand literacy practices in everyday use (e.g. Barton & Hamilton, 1998; Barton, Hamilton and Ivanic, 2000) including children’s appropriation of the media in school-based writing (Dyson, 1997). However, the changing semiotic and cultural practices associated with new media and online participation have less frequently been the object of study...