Epstein-Barr virus-positive diffuse large B-cell lymphoma of the elderly expresses EBNA3A with conserved CD8+ T-cell epitopes

Post-transplantation lymphoproliferative disorders (PTLD) arise in the immunosuppressed and are frequently Epstein-Barr virus (EBV) associated. The most common PTLD histological sub-type is diffuse large B-cell lymphoma (EBV+DLBCL-PTLD). Restoration of EBV-specific T-cell immunity can induce EBV+DLBCL-PTLD regression. The most frequent B-cell lymphoma in the immunocompetent is also DLBCL. ‘EBV-positive DLBCL of the elderly’ (EBV+DLBCL) is a rare but well-recognized DLBCL entity that occurs in the overtly immunocompetent, that has an adverse outcome relative to EBV-negative DLBCL. Unlike PTLD (which is classified as viral latency III), literature suggests EBV+DLBCL is typically latency II, i.e. expression is limited to the immuno-subdominant EBNA1, LMP1 and LMP2 EBV-proteins. If correct, this would be a major impediment for T-cell immunotherapeutic strategies. Unexpectedly we observed EBV+DLBCL-PTLD and EBV+DLBCL both shared features consistent with type III EBV-latency, including expression of the immuno-dominant EBNA3A protein. Extensive analysis showed frequent polymorphisms in EBNA1 and LMP1 functionally defined CD8+ T-cell epitope encoding regions, whereas EBNA3A polymorphisms were very rare making this an attractive immunotherapy target. As with EBV+DLBCL-PTLD, the antigen presenting machinery within lymphomatous nodes was intact. EBV+DLBCL express EBNA3A suggesting it is amenable to immunotherapeutic strategies.

The LCC15-MB human breast cancer cell line expresses osteopontin and exhibits an invasive and metastatic phenotype

We have characterized the LCC15-MB cell line which was recently derived from a breast carcinoma metastasis resected from the femur of a 29-year-old woman. LCC15-MB cells are vimentin (VIM) positive, exhibit a stellate morphology in routine cell culture, and form penetrating colonies when embedded in three-dimensional gels of Matrigel or fibrillar collagen. They show high levels of activity in the Boyden chamber chemomigration and chemoinvasion assays, and like other invasive human breast cancer (HBC) cell lines, LCC15-MB cells activate matrix-metalloproteinase-2 in response to treatment with concanavalin A. In addition, these cells are tumorigenic when implanted subcutaneously in nude mice and recolonize bone after arterial injection. Interestingly, both the primary lesion and the bone metastasis from which LCC15-MB were derived, as well as the resultant cell line, abundantly express the bone matrix protein osteopontin (OPN). OPN is also expressed by the highly metastatic MDA-MB-435 cells, but not other invasive or noninvasive HBC cell lines. Expression of OPN is retained in the subcutaneous xenograft and intraosseous metastases of LCC15-MB as detected by immunohistochemistry. Both VIM and OPN expression have been associated with breast cancer invasion and metastasis, and their expression by the LCC15-MB cell line is consistent with its derivation from a highly aggressive breast cancer. These cells provide a useful model for studying molecular mechanisms important for breast cancer metastasis to bone and, in particular, the implication(s) of OPN and VIM expression in this process.

Science education policy-making : eleven emerging issues

The Perth Declaration on Science and Technology Education of 2007 expresses strong concern about the state of science and technology education worldwide and calls on governments to respond to a number of suggestions for establishing the structural conditions for their improved practice. The quality of school education in science and technology has never before been of such critical importance to governments. There are three imperatives for its critical importance. The first relates to the traditional role of science in schooling, namely the identification, motivation and initial preparation of those students who will go on to further studies for careers in all those professional fi elds that directly involve science and technology. A suffi cient supply of these professionals is vital to the economy of all countries and to the health of their citizens. In the 21st century they are recognised everywhere as key players in ensuring that industrial and economic development occurs in a socially and environmentally sustainable way. In many countries this supply is now falling seriously short and urgently needs to be addressed. The second imperative is that sustainable technological development and many other possible societal applications of science require the support of scientifically and technologically informed citizens. Without the support and understanding of citizens, technological development can all too easily serve short term and sectional interests. The longer term progress of the whole society is overlooked, citizens will be confused about what should, and what should not be supported, and reactive and the environment will continue to be destroyed rather than sustained. Sustainable development, and the potential that science and technology increasingly offers, involves societies in ways that can often interact strongly, with traditional values, and hence, making decisions about them involve major moral decisions. All students need to be prepared through their science and technology education to be able to participate actively as persons and as responsible citizens in these essential and exciting possibilities. This goal is far from being generally achieved at present, but pathways to it are now more clearly understood. The third imperative derives from the changes that are resulting from the application of digital technologies that are the most rapid, the most widespread, and probably the most pervasive influence that science has ever had on human society. We all, wherever we live, are part of a global communication society. Information exchange and access to it that have been hitherto the realm of the few, are now literally in the hands of individuals. This is leading to profound changes in the World of Work and in what is known as the Knowledge Society. Schooling is now being challenged to contribute to the development in students of an active repertoire of generic and subject-based competencies. This contrasts very strongly with existing priorities, in subjects like the sciences that have seen the size of a student’s a store of established knowledge as the key measure of success. Science and technology education needs to be a key component in developing these competencies. When you add to these imperatives, the possibility that a more effective education in science and technology will enable more and more citizens to delight in, and feel a share in the great human enterprise we call Science, the case for new policy decisions is compellingly urgent. What follows are the recommendations (and some supplementary notes) for policy makers to consider about more operational aspects for improving science and technology education. They are listed under headings that point to the issues within each of these aspects. In the full document, a background is provided to each set of issues, including the commonly current state of science and technology education. Associated with each recommendation for consideration are the positive Prospects that could follow from such decision making, and the necessary Prerequisites, if such bold policy decisions are to fl ow, as intended, into practice in science and technology classrooms.

In-vehicle avatars to elicit social response and change driving behaviour

Social and psychological theories have provided a plethora of evidence showing that the physical difficulty to express appropriate social interactions between drivers expresses itself in aggression, selfish driving and anti-social behaviour. Therefore there is a need to improve interactions between drivers and allow clearer collective decision making between them. Personal characteristics and the driving situations play strong roles in driver’s aggression. Our approach is centered around the driving situation as opposed to focusing on personality characteristics. It examines aggression and manipulates contextual variables such as driver’s eye contact exchanges. This paper presents a new unobtrusive in-vehicle system that aims at communicating drivers’ intentions, elicit social responses and increasing mutual awareness. It uses eye gaze as a social cue to affect collective decision making with the view to contribute to safe driving. The authors used a driving simulator to design a case control experiment in which eye gaze movements are conveyed with an avatar. Participants were asked to drive through different types of intersections. An avatar representing the head of the other driver was displayed and driver behaviour was analysed. Significant eye gaze pattern difference where observed when an avatar was displayed. Drivers cautiously refer to the avatar when information is required on the intention of others (e.g. when they do not have the right of way). The majority of participants reported the perception of “being looked at”. The number of glances and time spent gazing at the avatar did not indicate an unsafe distraction by standards of in-vehicle device ergonomic design. Avatars were visually consulted primarily in less demanding driving situations, which underlines their non-distractive nature.

Enhancing learning in dance technique through on-line-mediated reflective practice

‘Practice makes perfect’ expresses the common misconception that repetitive practice without appropriate feed-back will deliver improvement in tasks being practised. This paper explores the implementation of a student-driven feed-back mechanism and shows how functional and aesthetic understanding can be progressively enhanced through reflective practice. More efficient practice of clearly understood tasks will enhance dance training outcomes. We were looking for ways to improve teaching efficiency, effectiveness of the students’ practice in the studio and application of safe dance practices. We devised a web-based on-line format, ‘Performing Reflective Practice’, designed to augment and refine studio practice. Only perfect practice makes perfect!

Hunger

Poet's statement: My father died of pancreatic cancer a few years ago, and since then other family members and friends have developed cancer. Some have recovered, perhaps temporarily, while for others the prospect is one of inevitable decline, raising questions about when the point is reached where death is preferable to life. This poem expresses the ambiguity of visceral urges which could be towards either continued life or a relieving death.

Analytical and numerical solutions for the time and space-symmetric fractional diffusion equation

We consider a time and space-symmetric fractional diffusion equation (TSS-FDE) under homogeneous Dirichlet conditions and homogeneous Neumann conditions. The TSS-FDE is obtained from the standard diffusion equation by replacing the first-order time derivative by a Caputo fractional derivative, and the second order space derivative by a symmetric fractional derivative. First, a method of separating variables expresses the analytical solution of the TSS-FDE in terms of the Mittag--Leffler function. Second, we propose two numerical methods to approximate the Caputo time fractional derivative: the finite difference method; and the Laplace transform method. The symmetric space fractional derivative is approximated using the matrix transform method. Finally, numerical results demonstrate the effectiveness of the numerical methods and to confirm the theoretical claims.

Empathy, neutrality and emotional intelligence: a balancing act for the emotional Einstein

Emotions play a central role in mediation as they help to define the scope and direction of a conflict. When a party to mediation expresses (and hence entrusts) their emotions to those present in a mediation, a mediator must do more than simply listen - they must attend to these emotions. Mediator empathy is an essential skill for communicating to a party that their feelings have been heard and understood, but it can lead mediators into trouble. Whilst there might exist a theoretical divide between the notions of empathy and sympathy, the very best characteristics of mediators (caring and compassionate nature) may see empathy and sympathy merge - resulting in challenges to mediator neutrality. This article first outlines the semantic difference between empathy and sympathy and the role that intrapsychic conflict can play in the convergence of these behavioural phenomena. It then defines emotional intelligence in the context of a mediation, suggesting that only the most emotionally intelligent mediators are able to emotionally connect with the parties, but maintain an impression of impartiality – the quality of remaining ‘attached yet detached’ to the process. It is argued that these emotionally intelligent mediators have the common qualities of strong self-awareness and emotional self-regulation.

Textbook portrayal of grieving stereotypes fails to cohere : a review

JANIS Balodis's Engine appears to have an educative purpose. Following the tragic death of her brother Stevie in a car crash, Natasha and her family struggle to cope with the devastation this sudden trauma has dealt them. Overlooked by her grieving parents, Natasha expresses her emotions by skipping school, self-harming and, as Engine unfolds, trying to enlist her Grumpop to help her finish rebuilding the car that could have saved Stevie's life by eliminating his need to catch a ride with a car full of friends. The symbolic action that drives Engine - rebuilding the car to rebuild Natasha and Grumpop's lives in the wake of trauma and guilt - is full of potential. It gives the design team, particularly designer Justin Nardella and composer Matt Hill, a strong premise to construct the garage that transforms into a home, a school, a church, a part, and the road on which the accident took place as the play unfolds.

A review of rationale for allocating costs and payments in producing and supplying public sector information PART A of 2 Parts – Chapters 1, 2 and 3) 13 March 2011 version

This work reviews the rationale and processes for raising revenue and allocating funds to perform information intensive activities that are pertinent to the work of democratic government. ‘Government of the people, by the people, for the people’ expresses an idea that democratic government has no higher authority than the people who agree to be bound by its rules. Democracy depends on continually learning how to develop understandings and agreements that can sustain voting majorities on which democratic law making and collective action depends. The objective expressed in constitutional terms is to deliver ‘peace, order and good government’. Meeting this objective requires a collective intellectual authority that can understand what is possible; and a collective moral authority to understand what ought to happen in practice. Facts of life determine that a society needs to retain its collective competence despite a continual turnover of its membership as people die but life goes on. Retaining this ‘collective competence’ in matters of self-government depends on each new generation: • acquiring a collective knowledge of how to produce goods and services needed to sustain a society and its capacity for self-government; • Learning how to defend society diplomatically and militarily in relation to external forces to prevent overthrow of its self-governing capacity; and • Learning how to defend society against divisive internal forces to preserve the authority of representative legislatures, allow peaceful dispute resolution and maintain social cohesion.

Morphine-induced receptor endocytosis in a novel knockin mouse reduces tolerance and dependence

Abstract Opioid drugs, such as morphine, are among the most effective analgesics available. However, their utility for the treatment of chronic pain is limited by side effects including tolerance and dependence. Morphine acts primarily through the mu-opioid receptor (MOP-R) , which is also a target of endogenous opioids. However, unlike endogenous ligands, morphine fails to promote substantial receptor endocytosis both in vitro, and in vivo. Receptor endocytosis serves at least two important functions in signal transduction. First, desensitization and endocytosis act as an "off" switch by uncoupling receptors from G protein. Second, endocytosis functions as an "on" switch, resensitizing receptors by recycling them to the plasma membrane. Thus, both the off and on function of the MOP-R are altered in response to morphine compared to endogenous ligands. To examine whether the low degree of endocytosis induced by morphine contributes to tolerance and dependence, we generated a knockin mouse that expresses a mutant MOP-R that undergoes morphine-induced endocytosis. Morphine remains an excellent antinociceptive agent in these mice. Importantly, these mice display substantially reduced antinociceptive tolerance and physical dependence. These data suggest that opioid drugs with a pharmacological profile similar to morphine but the ability to promote endocytosis could provide analgesia while having a reduced liability for promoting tolerance and dependence

Some reflections about P2M : the place of the mirror

The purpose of this paper is to provide some insights about P2M, and more specifically, to develop some thoughts about Project Management seen as a Mirror, a place for reflection…, between the Mission of organisation and its actual creation of Values (with s: a source of value for people, organisations and society). This place is the realm of complexity, of interactions between multiple variables, each of them having a specific time horizon and occupying a specific place, playing a specific role. Before developing this paper I would like to borrow to my colleague and friend, Professor Ohara, the following, part of a paper going to be presented at IPMA World Congress, in New Delhi later this year in November 2005. “P2M is the Japanese version of project & program management, which is the first standard guide for education and certification developed in 2001. A specific finding of P2M is characterized by “mission driven management of projects” or a program which harness complexity of problem solving observed in the interface between technical system and business model.” (Ohara, 2005, IPMA Conference, New Delhi) “The term of “mission” is a key word in the field of corporate strategy, where it expresses raison d’être or “value of business”. It is more specifically used for expressing “the client needs” in terms of a strategic business unit. The concept of mission is deemed to be a useful tool to share essential content of value and needs in message for complex project.” (Ohara, 2005, IPMA Conference, New Delhi) “Mission is considered as a significant “metamodel representation” by several reasons. First, it represents multiple values for aspiration. The central objective of mission initiative is profiling of ideality in the future from reality, which all stakeholders are glad to accept and share. Second, it shall be within a stretch of efforts, and not beyond or outside of the realization. Though it looks like unique, it has to depict a solid foundation. The pragmatic sense of equilibrium between innovation and adaptation is required for the mission. Third, it shall imply a rough sketch for solution to critical issues for problems in reality.” (Ohara, 2005, IPMA Conference, New Delhi) “Project modeling” idea has been introduced in P2M program management. A package of three project models of “scheme”, “system” and “service” are given as a reference type program. (Ohara, 2005, IPMA Conference, New Delhi) If these quotes apply to P2M, they are fully congruent with the results of the research undertaken and the resulting meta-model & meta-method developed by the CIMAP, ESC Lille Research Centre in Project & Program Management, since the 80’s. The paper starts by questioning the common Project Management (PM) paradigm. Then discussing the concept of Project, it argues that an alternative epistemological position should be taken to capture Page 2 / 11 the very nature of the PM field. Based on this, a development about “the need of modelling to understand” is proposed grounded on two theoretical roots. This leads to the conclusion that, in order to enables this modelling, a standard approach is necessary, but should be understood under the perspective of the Theory of Convention in order to facilitate a situational and contextual application.

Resourcing Early Learners : New Networks, New Actors

The landscape of early childhood education and care is changing. Governments world-wide are assuming increasing authority in relation to child-rearing in the years before school entry, beyond the traditional role in assisting parents to do the best they can by their children. As part of a social agenda aimed at forming citizens well prepared to play an active part in a globalised knowledge economy, the idea of ‘early learning’ expresses the necessity of engaging caregivers right from the start of children’s lives. Nichols, Rowsell, Rainbird, and Nixon investigate this trend over three years, in two countries, and three contrasting regions, by setting themselves the task of tracing every service and agent offering resources under the banner of early learning. Far from a dry catalogue, the study involves in-depth ethnographic research in fascinating spaces such as a church-run centre for African refugee women and children, a state-of-the-art community library and an Australian country town. Included is an unprecedented inventory of an entire suburban mall. Richly visually documented, the study employs emerging methods such as Google-mapping to trace the travels of actual parents as they search for particular resources. Each chapter features a context investigated in this large, international study: the library, the mall, the clinic, and the church. The author team unravels new spaces and new networks at work in early childhood literacy and development.

The roles of the preproghrelin-derived peptides - ghrelin, desacyl ghrelin and obestatin - in prostate cancer

Prostate cancer is the second most common cause of cancer related deaths in Western men. Despite the significant improvements in current treatment techniques, there is no cure for advanced metastatic, castrate-resistant disease. Early detection and prevention of progression to a castrate-resistant state may provide new strategies to improve survival. A number of growth factors have been shown to act in an autocrine/paracrine manner to modulate prostate cancer tumour growth. Our laboratory has previously shown that ghrelin and its receptors (the functional GHS-R1a and the non-functional GHS-R1b) are expressed in prostate cancer specimens and cell lines. We have shown that ghrelin increases cell proliferation in the PC3 and LNCaP prostate cancer cell lines through activation of ERK1/2, suggesting that ghrelin could regulate prostate cancer cell growth and play a role in the progression of the disease. Ghrelin is a 28 amino-acid peptide hormone, identified to be the natural ligand of the growth hormone secretagogue receptor (GHS-R1a). It is well characterised as a growth hormone releasing and as an orexigenic peptide that stimulates appetite and feeding and regulates energy expenditure and bodyweight. In addition to its orexigenic properties, ghrelin has been shown to play a regulatory role in a number of systems, including the reproductive, immune and cardiovascular systems and may play a role in a number of pathological conditions such as chronic heart failure, anorexia, cachexia, obesity, diabetes and cancer. In cancer, ghrelin and its receptor are expressed in a range of tumours and cancer cell lines and ghrelin has been demonstrated to modulate cell proliferation, apoptosis, migration and invasion in some cell types. The ghrelin gene (GHRL) encodes preproghrelin peptide, which is processed to produce three currently known functional peptides - ghrelin, desacyl ghrelin and obestatin. Prohormone convertases (PCs) have been shown to cleave the preproghrelin peptide into two primary products - the 28 amino acid peptide, ghrelin, and the remaining 117 amino acid C-terminal peptide, C-ghrelin. C-ghrelin can then be further processed to produce the 23 amino acid peptide, obestatin. Ghrelin circulates in two different forms - an octanoylated form (known as ghrelin) and a non-octanoylated form, desacyl ghrelin. The unique post-translational addition of octanoic acid to the serine 3 residue of the propeptide chain to form acylated ghrelin is catalysed by ghrelin O-acyltransferase (GOAT). This modification is necessary for binding of ghrelin to its only known functional receptor, the GHS-R1a. As desacyl ghrelin cannot bind and activate the GHS-R1a, it was initially thought to be an inactive peptide, despite the fact that it circulates at much higher levels than ghrelin. Further research has demonstrated that desacyl ghrelin is biologically active and shares some of the actions of ghrelin, as well as having some opposing and distinct roles. Interestingly, both ghrelin and desacyl ghrelin have been shown to modulate apoptosis, cell differentiation and proliferation in some cell types, and to stimulate cell proliferation through activation of ERK1/2 and PI3K/Akt pathways. The third known peptide product of the ghrelin preprohormone, obestatin, was initially thought to oppose the actions of ghrelin in appetite regulation and food intake and to mediate its effects through the G protein-coupled receptor 39 (GPR39). Subsequent research failed to reproduce the initial findings, however, and the possible anorexigenic effects of obestatin, as well as the identity of its receptor, remain unclear. Obestatin plays some important physiological roles, including roles in improving memory, the inhibition of thirst and anxiety, increased secretion of pancreatic juice, and regulation of cell proliferation, survival, apoptosis and differentiation. Preliminary studies have also shown that obestatin stimulates cell proliferation in some cell types through activation of ERK1/2, Akt and PKC pathways. Overall, however, at the commencement of this PhD project, relatively little was known regarding the functions and mechanisms of action of the preproghrelin-derived functional peptides in modulating prostate cancer cell proliferation. The roles of obestatin, and desacyl ghrelin as potential growth factors had not previously been investigated, and the potential expression and regulation of the preproghrelin processing enzymes, GOAT and prohormone convertases was unknown in prostate cancer cell lines. Therefore, the overall objectives of this study were to: 1. investigate the effects of obestatin on cell proliferation and signaling in prostate cancer cell lines 2. compare the effects of desacyl ghrelin and ghrelin on cell proliferation and signaling in prostate cancer cell lines 3. investigate whether prostate cancer cell lines possess the necessary enzymatic machinery to produce ghrelin and desacyl ghrelin and if these peptides can regulate GOAT expression Our laboratory has previously shown that ghrelin stimulates cell proliferation in the PC3 and LNCaP prostate cancer cell line through activation of the ERK1/2 pathway. In this study it has been demonstrated that treatments with either ghrelin, desacyl ghrelin or obestatin over 72 hours significantly increased cell proliferation in the PC3 prostate cancer cell line but had no significant effect in the RWPE-1 transformed normal prostate cell line. Ghrelin (1000nM) stimulated cell proliferation in the PC3 prostate cancer cell line by 31.66 6.68% (p<0.01) with the WST-1 method, and 13.55 5.68% (p<0.05) with the CyQUANT assay. Desacyl ghrelin (1000nM) increased cell proliferation in PC3 cells by 21.73 2.62% (p<0.01) (WST-1), and 15.46 7.05% (p<0.05) (CyQUANT) above untreated control. Obestatin (1000nM) induced a 28.37 7.47% (p<0.01) (WST-1) and 12.14 7.47% (p<0.05) (CyQUANT) significant increase in cell proliferation in the PC3 prostate cancer cell line. Ghrelin and desacyl ghrelin treatments stimulated Akt and ERK phosphorylation across a range of concentrations (p<0.01). Obestatin treatment significantly stimulated Akt, ERK and PKC phosphorylation (p<0.05). Through the use of specific inhibitors, the MAPK inhibitor U0126 and the Akt1/2 kinase inhibitor, it was demonstrated that ghrelin- and obestatin-induced cell proliferation in the PC3 prostate cancer cell line is mediated through activation of ERK1/2 and Akt pathways. Although desacyl ghrelin significantly stimulated Akt and ERK phosphorylation, U0126 failed to prevent desacyl ghrelin-induced cell proliferation suggesting ghrelin and desacyl ghrelin might act through different mechanisms to increase cell proliferation. Ghrelin and desacyl ghrelin have shown a proliferative effect in osteoblasts, pancreatic -cells and cardiomyocytes through activation of ERK1/2 and PI3K/Akt pathways. Here it has been shown that ghrelin and its non-acylated form exert the same function and stimulate cell proliferation in the PC3 prostate cancer cell line through activation of the Akt pathway. Ghrelin-induced proliferation was also mediated through activation of the ERK1/2 pathway, however, desacyl ghrelin seems to stimulate cell proliferation in an ERK1/2-independent manner. As desacyl ghrelin does not bind and activate GHSR1a, the only known functional ghrelin receptor, the finding that both ghrelin and desacyl ghrelin stimulate cell proliferation in the PC3 cell line suggests that these peptides could be acting through the yet unidentified alternative ghrelin receptor in this cell type. Obestatin treatment also stimulated PKC phosphorylation, however, a direct role for this pathway in stimulating cell proliferation could not be proven using available PKC pathway inhibitors, as they caused significant cell death over the extended timeframe of the cell proliferation assays. Obestatin has been shown to stimulate cell proliferation through activation of PKC isoforms in human retinal epithelial cells and in the human gastric cancer cell line KATO-III. We have demonstrated that all of the prostate-derived cell lines examined (PC3, LNCaP, DU145, 22Rv1, RWPE-1 and RWPE-2) expressed GOAT and at least one of the prohormone convertases, which are known to cleave the proghrelin peptide, PC1/3, PC2 and furin, at the mRNA level. These cells, therefore, are likely to possess the necessary machinery to cleave the preproghrelin protein and to produce the mature ghrelin and desacyl ghrelin peptides. In addition to prohormone convertases, the presence of octanoic acid is essential for acylated ghrelin production. In this study octanoic acid supplementation significantly increased cell proliferation in the PC3 prostate cancer cell line by over 20% compared to untreated controls (p<0.01), but surprisingly, not in the DU145, LNCaP or 22Rv1 prostate cancer cell lines or in the RWPE-1 and RWPE-2 prostate-derived cell lines. In addition, we demonstrated that exogenous ghrelin induced a statistically significant two-fold decrease in GOAT mRNA expression in the PC3 cell line (p<0.05), suggesting that ghrelin could pontentially downregulate its own acylation and, therefore, regulate the balance between ghrelin and desacyl ghrelin. This was not observed, however, in the DU145 and LNCaP prostate cancer cell lines. The GOAT-ghrelin system represents a direct link between ingested nutrients and regulation of ghrelin production and the ghrelin/desacyl ghrelin ratio. Regulation of ghrelin acylation is a potentially attractive and desirable tool for the development of better therapies for a number of pathological conditions where ghrelin has been shown to play a key role. The finding that desacyl ghrelin stimulates cell proliferation in the PC3 prostate cancer cell line, and responds to ghrelin in the same way, suggests that this cell line expresses an alternative ghrelin receptor. Although all the cell lines examined expressed both GHS-R1a and GHS-R1b mRNA, it remains uncertain whether these cell lines express the unidentified alternative ghrelin receptor. It is possible that the varied responses seen could be due to the expression of different ghrelin receptors in different cell lines. In addition to GOAT, prohormone convertases and octanoic acid availability may regulate the production of different peptides from the ghrelin preprohormone. The studies presented in this thesis provide significant new information regarding the roles and mechanisms of action of the preproghrelin-derived peptides, ghrelin, desacyl ghrelin and obestatin, in modulating prostate cancer cell line proliferation. A number of key questions remain to be resolved, however, including the identification of the alternative ghrelin/desacyl ghrelin receptor, the identification of the obestatin receptor, a clarification of the signaling mechanisms which mediate cell proliferation in response to obestatin treatment and a better understanding of the regulation at both the gene and post-translational levels of functional peptide generation. Further studies investigating the role of the ghrelin axis using in vivo prostate cancer models may be warranted. Until these issues are determined, the potential for the ghrelin axis, to be recognised as a novel useful target for therapy for cancer or other pathologies will be uncertain.