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.

Fast and accurate phonetic spoken term detection

For the first time in human history, large volumes of spoken audio are being broadcast, made available on the internet, archived, and monitored for surveillance every day. New technologies are urgently required to unlock these vast and powerful stores of information. Spoken Term Detection (STD) systems provide access to speech collections by detecting individual occurrences of specified search terms. The aim of this work is to develop improved STD solutions based on phonetic indexing. In particular, this work aims to develop phonetic STD systems for applications that require open-vocabulary search, fast indexing and search speeds, and accurate term detection. Within this scope, novel contributions are made within two research themes, that is, accommodating phone recognition errors and, secondly, modelling uncertainty with probabilistic scores. A state-of-the-art Dynamic Match Lattice Spotting (DMLS) system is used to address the problem of accommodating phone recognition errors with approximate phone sequence matching. Extensive experimentation on the use of DMLS is carried out and a number of novel enhancements are developed that provide for faster indexing, faster search, and improved accuracy. Firstly, a novel comparison of methods for deriving a phone error cost model is presented to improve STD accuracy, resulting in up to a 33% improvement in the Figure of Merit. A method is also presented for drastically increasing the speed of DMLS search by at least an order of magnitude with no loss in search accuracy. An investigation is then presented of the effects of increasing indexing speed for DMLS, by using simpler modelling during phone decoding, with results highlighting the trade-off between indexing speed, search speed and search accuracy. The Figure of Merit is further improved by up to 25% using a novel proposal to utilise word-level language modelling during DMLS indexing. Analysis shows that this use of language modelling can, however, be unhelpful or even disadvantageous for terms with a very low language model probability. The DMLS approach to STD involves generating an index of phone sequences using phone recognition. An alternative approach to phonetic STD is also investigated that instead indexes probabilistic acoustic scores in the form of a posterior-feature matrix. A state-of-the-art system is described and its use for STD is explored through several experiments on spontaneous conversational telephone speech. A novel technique and framework is proposed for discriminatively training such a system to directly maximise the Figure of Merit. This results in a 13% improvement in the Figure of Merit on held-out data. The framework is also found to be particularly useful for index compression in conjunction with the proposed optimisation technique, providing for a substantial index compression factor in addition to an overall gain in the Figure of Merit. These contributions significantly advance the state-of-the-art in phonetic STD, by improving the utility of such systems in a wide range of applications.

Obituary - Rigid contact lenses

Scleral and corneal rigid lenses represented 100 per cent of the contact lens market immediately prior to the invention of soft lenses in the mid-1960s. In the United Kingdom today, rigid lenses comprise 2 per cent of all new lens fits. Low rates of rigid lens fitting are also apparent in 27 other countries which have recently been surveyed. Thus, the 1998 prediction of the author that rigid lenses – also referred to as ‘rigid gas permeable’ (RGP) lenses or ‘gas permeable’ (GP) lenses – would be obsolete by the year 2010 has essentially turned out to be correct. In this obituary, the author offers 10 reasons for the demise of rigid lens fitting: initial rigid lens discomfort; intractable rigid lens-induced corneal and lid pathology; extensive soft lens advertising; superior soft lens fitting logistics; lack of rigid lens training opportunities; redundancy of the rigid lens ‘problem solver’ function; improved soft toric and bifocal/varifocal lenses; limited uptake of orthokeratology; lack of investment in rigid lenses; and the emergence of aberration control soft lenses. Rigid lenses are now being fitted by a minority of practitioners with specialist skills/training. Certainly, rigid lenses can no longer be considered as a mainstream form of contact lens correction. May their dear souls (bulk properties) rest in peace.

An examination of factors influencing effective and continuous improvement of Indonesian contractors' quality management systems

This paper presents the results of a pilot study examining the factors that impact most on the effective implementation of, and improvement to, Quality Mangement Sytems (QMSs) amongst Indonesian construction companies. Nine critical factors were identified from an extensive literature review, and a survey was conducted of 23 respondents from three specific groups (Quality Managers, Project Managers, and Site Engineers) undertaking work in the Indonesian infrastructure construction sector. The data has been analyzed initially using simple descriptive techniques. This study reveals that different groups within the sector have different opinions of the factors regardless of the degree of importance of each factor. However, the evaluation of construction project success and the incentive schemes for high performance staff, are the two factors that were considered very important by most of the respondents in all three groups. In terms of their assessment of tools for measuring contractor’s performance, additional QMS guidelines, techniques related to QMS practice provided by the Government, and benchmarking, a clear majority in each group regarded their usefulness as ‘of some importance’.

An enriched radial point interpolation method (e-RPIM) for analysis of crack tip fields

In this paper, an enriched radial point interpolation method (e-RPIM) is developed the for the determination of crack tip fields. In e-RPIM, the conventional RBF interpolation is novelly augmented by the suitable trigonometric basis functions to reflect the properties of stresses for the crack tip fields. The performance of the enriched RBF meshfree shape functions is firstly investigated to fit different surfaces. The surface fitting results have proven that, comparing with the conventional RBF shape function, the enriched RBF shape function has: (1) a similar accuracy to fit a polynomial surface; (2) a much better accuracy to fit a trigonometric surface; and (3) a similar interpolation stability without increase of the condition number of the RBF interpolation matrix. Therefore, it has proven that the enriched RBF shape function will not only possess all advantages of the conventional RBF shape function, but also can accurately reflect the properties of stresses for the crack tip fields. The system of equations for the crack analysis is then derived based on the enriched RBF meshfree shape function and the meshfree weak-form. Several problems of linear fracture mechanics are simulated using this newlydeveloped e-RPIM method. It has demonstrated that the present e-RPIM is very accurate and stable, and it has a good potential to develop a practical simulation tool for fracture mechanics problems.

Tendering theory revisited

This paper discusses the content, origin and development of Tendering Theory as a theory of price determination. It demonstrates how tendering theory determines market prices and how it is different from game and decision theories, and that in the tendering process, with non-cooperative, simultaneous, single sealed bids with individual private valuations, extensive public information, a large number of bidders and a long sequence of tendering occasions, there develops a competitive equilibrium. The development of a competitive equilibrium means that the concept of the tender as the sum of a valuation and a strategy, which is at the core of tendering theory, cannot be supported and that there are serious empirical, theoretical and methodological inconsistencies in the theory.

Effectiveness of constructability concept in the provision of infrastructure assets

The concept of constructability is to use construction knowledge and experience during all phases of a project, particularly in the earliest phases of planning and design. It facilitates project objectives before delivery stage, and decreases unnecessary costs during construction phase. Despite the extensive use, constructability concept fails to address many issues related to Operation and Maintenance (O&M) of construction projects. Extending constructability concept, to include the O&M issues, could lead to the projects that are not fitted for construction purposes only, but also fitted for use. This study reviews the literature of constructability implementation, its benefits and shortcomings during the infrastructure life cycle, as well as the delivery success factors of infrastructure projects. This contributes to the propose need of a model to improve the effectiveness and efficiency of infrastructure project by extending the concept of constructability to include O&M. Development of such a model can facilitate post-occupancy stakeholders’ participation in a constructability program. It will help infrastructure owners eliminate project reworks, and improve O&M effectiveness and efficiency.

Model-order selection in Zernike polynomial expansion of corneal surfaces using the efficient detection criterion

Corneal-height data are typically measured with videokeratoscopes and modeled using a set of orthogonal Zernike polynomials. We address the estimation of the number of Zernike polynomials, which is formalized as a model-order selection problem in linear regression. Classical information-theoretic criteria tend to overestimate the corneal surface due to the weakness of their penalty functions, while bootstrap-based techniques tend to underestimate the surface or require extensive processing. In this paper, we propose to use the efficient detection criterion (EDC), which has the same general form of information-theoretic-based criteria, as an alternative to estimating the optimal number of Zernike polynomials. We first show, via simulations, that the EDC outperforms a large number of information-theoretic criteria and resampling-based techniques. We then illustrate that using the EDC for real corneas results in models that are in closer agreement with clinical expectations and provides means for distinguishing normal corneal surfaces from astigmatic and keratoconic surfaces.

Nursing and Midwifery Services Strategic Directions, 2011-2015

This document presents the newly updated strategic directions for strengthening nursing and midwifery services (SDNM) for the period 2011–2015. Complementing and building on the 2002–2008 SDNM, it seeks to provide policymakers, practitioners and other stakeholders at every level with a flexible framework for broad-based, collaborative action to enhance the capacity of nurses and midwives to contribute to: * universal coverage * people-centred health care * policies affecting their practice and working conditions, and the * scaling up of national health systems to meet global goals and targets. The SDNM for 2011–2015 draws on several key World Health Assembly resolutions, and are underpinned by the associated global policy recommendations and codes of practice. (1,2) After two years of extensive research and consultation, a SDNM task force was developed, and a consensus on a range of specific activities revolving around 13 objectives in five interrelated key results areas (KRAs), was achieved: n health system and service strengthening n policy and practice * education, training and career development * workforce management and * partnership. Stakeholders, although free to prioritize certain parts of the framework to meet their own particular needs, are encouraged to adhere to the cornerstone of collaborative action, namely the common goal enshrined in the core SDNM 2011–2015 vision statement: improved health outcomes for individuals, families and communities through the provision of competent, culturally sensitive, evidence-based nursing and midwifery services.

Enhancing Water Sensitive Urban Design (WSUD) practices to mitigate urban stormwater pollution and reuse potential

Water Sensitive Urban Design (WSUD) practices such as wetlands, bioretention systems and swales are widely implemented in Australia’s urban areas for the mitigation of stormwater pollution and to enhance its reuse potential. In-depth research undertaken has confirmed that these systems do not always perform according to design expectations due to a diversity of reasons. To deliver anticipated benefits, it is critical that they are designed in conformity with catchment and rainfall characteristics and pollutant processes. This in turn entails an in-depth understanding of key pollutant processes. This paper presents the outcomes of extensive research investigations on pollutant characterisation and stormwater pollutant processes on urban catchment surfaces. Outcomes from the research studies revealed the complexities in physical and chemical characteristics of pollutants originating from urban catchments which are strongly influenced by rainfall and catchment characteristics. Based on the research outcomes, recommendations are provided to enhance stormwater treatment performance and to enhance its reuse potential.

A coupled meshless technique/molecular dynamics approach for deformation characterization of monocrystalline metal

This paper presents a multiscale study using the coupled Meshless technique/Molecular Dynamics (M2) for exploring the deformation mechanism of mono-crystalline metal (focus on copper) under uniaxial tension. In M2, an advanced transition algorithm using transition particles is employed to ensure the compatibility of both displacements and their gradients, and an effective local quasi-continuum approach is also applied to obtain the equivalent continuum strain energy density based on the atomistic poentials and Cauchy-Born rule. The key parameters used in M2 are firstly investigated using a benchmark problem. Then M2 is applied to the multiscale simulation for a mono-crystalline copper bar. It has found that the mono-crystalline copper has very good elongation property, and the ultimate strength and Young's modulus are much higher than those obtained in macro-scale.

Planning and implementing trajectories for autonomous underwater vehicles to track evolving ocean processes based on predictions from a regional ocean model

Path planning and trajectory design for autonomous underwater vehicles (AUVs) is of great importance to the oceanographic research community because automated data collection is becoming more prevalent. Intelligent planning is required to maneuver a vehicle to high-valued locations to perform data collection. In this paper, we present algorithms that determine paths for AUVs to track evolving features of interest in the ocean by considering the output of predictive ocean models. While traversing the computed path, the vehicle provides near-real-time, in situ measurements back to the model, with the intent to increase the skill of future predictions in the local region. The results presented here extend prelim- inary developments of the path planning portion of an end-to-end autonomous prediction and tasking system for aquatic, mobile sensor networks. This extension is the incorporation of multiple vehicles to track the centroid and the boundary of the extent of a feature of interest. Similar algorithms to those presented here are under development to consider additional locations for multiple types of features. The primary focus here is on algorithm development utilizing model predictions to assist in solving the motion planning problem of steering an AUV to high-valued locations, with respect to the data desired. We discuss the design technique to generate the paths, present simulation results and provide experimental data from field deployments for tracking dynamic features by use of an AUV in the Southern California coastal ocean.

Cooperative multi-AUV tracking of phytoplankton blooms based on ocean model predictions

In recent years, ocean scientists have started to employ many new forms of technology as integral pieces in oceanographic data collection for the study and prediction of complex and dynamic ocean phenomena. One area of technological advancement in ocean sampling if the use of Autonomous Underwater Vehicles (AUVs) as mobile sensor plat- forms. Currently, most AUV deployments execute a lawnmower- type pattern or repeated transects for surveys and sampling missions. An advantage of these missions is that the regularity of the trajectory design generally makes it easier to extract the exact path of the vehicle via post-processing. However, if the deployment region for the pattern is poorly selected, the AUV can entirely miss collecting data during an event of specific interest. Here, we consider an innovative technology toolchain to assist in determining the deployment location and executed paths for AUVs to maximize scientific information gain about dynamically evolving ocean phenomena. In particular, we provide an assessment of computed paths based on ocean model predictions designed to put AUVs in the right place at the right time to gather data related to the understanding of algal and phytoplankton blooms.

Autonomous underwater vehicle trajectory design coupled with predictive ocean models : a case study

Data collection using Autonomous Underwater Vehicles (AUVs) is increasing in importance within the oceano- graphic research community. Contrary to traditional moored or static platforms, mobile sensors require intelligent planning strategies to manoeuvre through the ocean. However, the ability to navigate to high-value locations and collect data with specific scientific merit is worth the planning efforts. In this study, we examine the use of ocean model predictions to determine the locations to be visited by an AUV, and aid in planning the trajectory that the vehicle executes during the sampling mission. The objectives are: a) to provide near-real time, in situ measurements to a large-scale ocean model to increase the skill of future predictions, and b) to utilize ocean model predictions as a component in an end-to-end autonomous prediction and tasking system for aquatic, mobile sensor networks. We present an algorithm designed to generate paths for AUVs to track a dynamically evolving ocean feature utilizing ocean model predictions. This builds on previous work in this area by incorporating the predicted current velocities into the path planning to assist in solving the 3-D motion planning problem of steering an AUV between two selected locations. We present simulation results for tracking a fresh water plume by use of our algorithm. Additionally, we present experimental results from field trials that test the skill of the model used as well as the incorporation of the model predictions into an AUV trajectory planner. These results indicate a modest, but measurable, improvement in surfacing error when the model predictions are incorporated into the planner.