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.

An evolution of waterfront development in Malaysia

River has long been recognized as one of humanity’s most important natural resources. It is one of the most important of all the natural resources necessary to ensure human health and civilization. A close association between cities and water is inherent since the history of civilization and in fact, many urban cities in Malaysia are located close to river areas. The last two decades shown Malaysia has shifted development strategy from agricultural based to industrialization, and manufacturing industries have become the economy’s main source for the country until now. This transformation in 18th century is clearly shown that rapid urbanization, industrial and intensive agricultural activities, as well as wide-spread land development, have contributed to extensive changing of river functions for economy, national development and environment. In particular, river roles are become less significance for human life and river function limited only for transportation purposes only. So, viewed historically, waterfront development in Malaysia have undergone cycles of change over the decades and the latest in this pattern to more public purposes such as recreational and mixed used development. This paper aims to identify a transition of waterfront development in Malaysia from history time to modernization era and it would give a significance contribution for the research is currently on going.

Preface : rethinking sustainable development : urban management, engineering, and design

In an atmosphere where civilization is progressing and becoming more aware of the consequences of careless development decisions, rethinking sustainable development - particularly sustainable urban and infrastructure development - has become an inevitable necessity. ------ ----- Rethinking Sustainable Development: Urban Management, Engineering, and Design considers the role of urban, regional and infrastructure planning in achieving sustainable urban and infrastructure development, providing insights into overcoming the consequences of unsustainable development. This companion volume to Sustainable Urban and Regional Infrastructure: Technology, Planning and Management, overviews all aspects of sustainable urban and infrastructure development.

Ecological sustainable development: a study of the obstacles in Australia's construction industry

Ecological sustainable development (ESD), defined as that which meets the needs of the present without compromising the ability of future generations to meet their own needs, has much to offer in enhancing the quality of life of people and maintaining the environment for future generations by reducing the pollution of water, air and land, minimizing the destruction of irreplaceable ecosystems and cutting down the amount of toxic materials released. However, there is still much to do to achieve full implementation world-wide. This paper reports on three factors-design, attitudes and financial constraints - that are likely barriers to the implementation of ESD within the built environment in Australian industry. A postal questionnaire survey is described aimed at soliciting views on detailed aspects of the factors. This shows that ESD in the Australian built environment has also not been successfully implemented. The main reason is found to be due to the perceived costs involved - the cost of using environmental materials being a predominant factor. The design of ESD, being more sophisticated, also is perceived as involving stakeholders in more expense. There also appears to be a lack of knowledge and a lack of specialised and interdisciplinary design teams available in the Australian context.

"Criatividade na Educação e Formação de Talentos" (Creativity in Education and Development of Talent)

Historically, 'Creativity' has had a complex set of meanings. Not long ago, 'Creativity' had a kind of marginal or peripheral status, being seen as the province of a gifted few; in many cases it was associated almost exclusively with the arts and with artists. But these traditional attitudes to creativity are changing. Mainstream businesses are employing people with creative skills as diverse as writing, directing, graphic design and event management. So what we’re beginning to see is an innovation framework and creative content adding value not just to SMEs, but to traditional industries such as manufacturing and mining, and to wider service industries. And this is why Education is such an important element, particularly with a focus on innovation, and on creative people and the contributions they make across different parts of the economy.

A measurement instrument for process modeling research : development, test and procedural model

Process modeling is an emergent area of Information Systems research that is characterized through an abundance of conceptual work with little empirical research. To fill this gap, this paper reports on the development and validation of an instrument to measure user acceptance of process modeling grammars. We advance an extended model for a multi-stage measurement instrument development procedure, which incorporates feedback from both expert and user panels. We identify two main contributions: First, we provide a validated measurement instrument for the study of user acceptance of process modeling grammars, which can be used to assist in further empirical studies that investigate phenomena associated with the business process modeling domain. Second, in doing so, we describe in detail a procedural model for developing measurement instruments that ensures high levels of reliability and validity, which may assist fellow scholars in executing their empirical research.

Sustainable urban and regional infrastructure development : technologies, applications and management

Sustainable Urban and Regional Infrastructure Development: Technologies, Applications and Management, bridges the gap in the current literature by addressing the overall problems present in society's major infrastructures, and the technologies that may be applied to overcome these problems. It focuses on ways in which energy intensive but 'invisible' (to the general public) facilities can become green or greener. The studies presented re lessons to be learnt from our neighbors and from our own backyard, and provide an excellent general overview of the issues facing us all.

Knowledge-based development for cities and societies : integrated multi-level approaches

Over the past twenty years, the conventional knowledge management approach has evolved into a strategic management approach that has found applications and opportunities outside of business, in society at large, through education, urban development, governance, and healthcare, amongst others. Knowledge-Based Development for Cities and Socieities: Integrated Multi-Level Approaches enlightens the concepts and challenges of knowledge management for both urban environments and entire regions, enhancing the expertise and knowledge of scholars, resdearchers, practitioners, managers and urban developers in the development of successful knowledge-based development policies, creation of knowledte cities and prosperous knowledge societies. This reference creates large knowledge base for scholars, managers and urban developers and increases the awareness of the role of knowledge cities and knowledge socieiteis in the knowledge era, as well as of the challenges and opportunities for future research.

Executive function effects and numerical development in children : behavioural and ERP evidence from a numerical Stroop paradigm

Most research on numerical development in children is behavioural, focusing on accuracy and response time in different problem formats. However, Temple and Posner (1998) used ERPs and the numerical distance task with 5-year-olds to show that the development of numerical representations is difficult to disentangle from the development of the executive components of response organization and execution. Here we use the numerical Stroop paradigm (NSP) and ERPs to study possible executive interference in numerical processing tasks in 6–8-year-old children. In the NSP, the numerical magnitude of the digits is task-relevant and the physical size of the digits is task-irrelevant. We show that younger children are highly susceptible to interference from irrelevant physical information such as digit size, but that access to the numerical representation is almost as fast in young children as in adults. We argue that the developmental trajectories for executive function and numerical processing may act together to determine numerical development in young children.

An evidence-based approach to professional development planning in large state educational systems

This paper presents a systems-level approach for adjudicating the prioritization, selection, and planning of inservcie professional development (PD) for teachers. We present a step-by-step model for documenting and assessing system-wide 'bids' for professional development programs

Developing a knowledge-based urban development analysis framework : the case of multimedia super corridor, Malaysia

Purpose: In the global knowledge economy, investment in knowledge-intensive industries and information and communication technology (ICT) infrastructures are seen as significant factors in improving the overall socio-economic fabric of cities. Consequently knowledge-based urban development (KBUD) has become a new paradigm in urban planning and development, for increasing the welfare and competitiveness of cities and regions. The paper discusses the critical connections between KBUD strategies and knowledge-intensive industries and ICT infrastructures. In particular, it investigates the application of the knowledge-based urban development concept by discussing one of South East Asia’s large scale manifestations of KBUD; Malaysia’s Multimedia Super Corridor. ----- ----- Design/methodology/approach: The paper provides a review of the KBUD concept and develops a knowledge-based urban development assessment framework to provide a clearer understanding of development and evolution of KBUD manifestations. Subsequently the paper investigates the implementation of the KBUD concept within the Malaysian context, and particularly the Multimedia Super Corridor (MSC). ----- ----- Originality/value: The paper, with its KBUD assessment framework, scrutinises Malaysia’s experince; providing an overview of the MSC project and discussion of the case findings. The development and evolution of the MSC is viewed with regard to KBUD policy implementation, infrastructural implications, and the agencies involved in the development and management of the MSC. ----- ----- Practical implications: The emergence of the knowledge economy, together with the issues of globalisation and rapid urbanisation, have created an urgent need for urban planners to explore new ways of strategising planning and development that encompasses the needs and requirements of the knowledge economy and society. In light of the literature and MSC case findings, the paper provides generic recommendations, on the orchestration of knowledge-based urban development, for other cities and regions seeking to transform to the knowledge economy.

Editorial : knowledge-based development of cities : a myth or reality?

Urban development in the first decade of the 21st century has faced many challenges ranging from rapid to shrinking urbanisation, from emerging knowledge economy to global division of labour and from globalisation to climate change. Along with these challenges new concepts, such as essentialism, environmentalism and dematerialism, are emerged and started to influence the way urban development plans are prepared and visions for the development of cities are made. Beyond this, scholars, practitioners and decision-makers have also started to discuss the need for an new urban planning and development approach in order to achieve a development that is sustainable and knowledge-based. Limited successful examples of alternative planning and development approaches showcased potentials of moving towards a new plan-making mindset in the era of knowledge economy. This paper presents a new urban planning and development approach that is taking application ground in many parts of the globe, namely knowledge-based urban development. After providing the theoretical foundation and conceptual framework of knowledge-based urban development the paper discusses whether knowledge-based development of cities is a myth or a reality.

Autonomous underwater vehicles: development and implementation of time and energy efficient trajectories

Autonomous underwater vehicles (AUVs) are increasingly used, both in military and civilian applications. These vehicles are limited mainly by the intelligence we give them and the life of their batteries. Research is active to extend vehicle autonomy in both aspects. Our intent is to give the vehicle the ability to adapt its behavior under different mission scenarios (emergency maneuvers versus long duration monitoring). This involves a search for optimal trajectories minimizing time, energy or a combination of both. Despite some success stories in AUV control, optimal control is still a very underdeveloped area. Adaptive control research has contributed to cost minimization problems, but vehicle design has been the driving force for advancement in optimal control research. We look to advance the development of optimal control theory by expanding the motions along which AUVs travel. Traditionally, AUVs have taken the role of performing the long data gathering mission in the open ocean with little to no interaction with their surroundings, MacIver et al. (2004). The AUV is used to find the shipwreck, and the remotely operated vehicle (ROV) handles the exploration up close. AUV mission profiles of this sort are best suited through the use of a torpedo shaped AUV, Bertram and Alvarez (2006), since straight lines and minimal (0 deg - 30 deg) angular displacements are all that are necessary to perform the transects and grid lines for these applications. However, the torpedo shape AUV lacks the ability to perform low-speed maneuvers in cluttered environments, such as autonomous exploration close to the seabed and around obstacles, MacIver et al. (2004). Thus, we consider an agile vehicle capable of movement in six degrees of freedom without any preference of direction.

Preface : sustainable urban and regional infrastructure development : technologies, applications and management

The concept of ‘sustainability’ has been pushed to the forefront of policy-making and politics as the world wakes up to the impacts of climate change and the effects of the modern urban lifestyle. Climate change has emerged to be one of the biggest challenges faced by our planet today, threatening both built and natural systems with long term consequences which may be irreversible. While there is a vast literature in the market on sustainable cities and urban development, there is currently none that bring together the vital issues of urban and regional development, and the planning, management and implementation of sustainable infrastructure. Large scale infrastructure plays an important part in modern society by not only promoting economic growth, but also by acting as a key indicator for it. More importantly, it supplies municipal/local amenity and services: water, electricity, social and communication facilities, waste removal, transport of people and goods, as well as numerous other services. For the most part, infrastructure has been built by teams lead by engineers who are more concerned about functionality than the concept of sustainability. However, it has been widely stated that current practices and lifestyle cannot continue if we are to leave a healthy living planet to not only the next generation, but also to the generations beyond. Therefore, in order to be sustainable, there are drastic measures that need to be taken. Current single purpose and design infrastructures that are open looped are not sustainable; they are too resource intensive, consume too much energy and support the consumption of natural resources at a rate that will exhaust their supply. Because of this, it is vital that modern society, policy-makers, developers, engineers and planners become pioneers in introducing and incorporating sustainable features into urban and regional infrastructure.