Field Testing and Development of Novel SHM systems at Queen’s University Belfast

Ageing and deterioration of infrastructure is a challenge facing transport authorities. In
particular, there is a need for increased bridge monitoring in order to provide adequate
maintenance and to guarantee acceptable levels of transport safety. The Intelligent
Infrastructure group at Queens University Belfast (QUB) are working on a number of aspects
of infrastructure monitoring and this paper presents summarised results from three distinct
monitoring projects carried out by this group. Firstly the findings from a project on next
generation Bridge Weight in Motion (B-WIM) are reported, this includes full scale field testing
using fibre optic strain sensors. Secondly, results from early phase testing of a computer
vision system for bridge deflection monitoring are reported on. This research seeks to exploit
recent advances in image processing technology with a view to developing contactless
bridge monitoring approaches. Considering the logistical difficulty of installing sensors on a
‘live’ bridge, contactless monitoring has some inherent advantages over conventional
contact based sensing systems. Finally the last section of the paper presents some recent
findings on drive by bridge monitoring. In practice a drive-by monitoring system will likely
require GPS to allow the response of a given bridge to be identified; this study looks at the
feasibility of using low-cost GPS sensors for this purpose, via field trials. The three topics
outlined above cover a spectrum of SHM approaches namely, wired monitoring, contactless
monitoring and drive by monitoring

New methods for movement technique development in cross-country skiing using mathematical models and simulation

This Licentiate Thesis is devoted to the presentation and discussion of some new contributions in applied mathematics directed towards scientific computing in sports engineering. It considers inverse problems of biomechanical simulations with rigid body musculoskeletal systems especially in cross-country skiing. This is a contrast to the main research on cross-country skiing biomechanics, which is based mainly on experimental testing alone. The thesis consists of an introduction and five papers. The introduction motivates the context of the papers and puts them into a more general framework. Two papers (D and E) consider studies of real questions in cross-country skiing, which are modelled and simulated. The results give some interesting indications, concerning these challenging questions, which can be used as a basis for further research. However, the measurements are not accurate enough to give the final answers. Paper C is a simulation study which is more extensive than paper D and E, and is compared to electromyography measurements in the literature. Validation in biomechanical simulations is difficult and reducing mathematical errors is one way of reaching closer to more realistic results. Paper A examines well-posedness for forward dynamics with full muscle dynamics. Moreover, paper B is a technical report which describes the problem formulation and mathematical models and simulation from paper A in more detail. Our new modelling together with the simulations enable new possibilities. This is similar to simulations of applications in other engineering fields, and need in the same way be handled with care in order to achieve reliable results. The results in this thesis indicate that it can be very useful to use mathematical modelling and numerical simulations when describing cross-country skiing biomechanics. Hence, this thesis contributes to the possibility of beginning to use and develop such modelling and simulation techniques also in this context.

Development of Imaging Mass Spectrometry Analysis of Lipids in Biological and Clinically Relevant Applications

La spectrométrie de masse mesure la masse des ions selon leur rapport masse sur charge. Cette technique est employée dans plusieurs domaines et peut analyser des mélanges complexes. L’imagerie par spectrométrie de masse (Imaging Mass Spectrometry en anglais, IMS), une branche de la spectrométrie de masse, permet l’analyse des ions sur une surface, tout en conservant l’organisation spatiale des ions détectés. Jusqu’à présent, les échantillons les plus étudiés en IMS sont des sections tissulaires végétales ou animales. Parmi les molécules couramment analysées par l’IMS, les lipides ont suscité beaucoup d'intérêt. Les lipides sont impliqués dans les maladies et le fonctionnement normal des cellules; ils forment la membrane cellulaire et ont plusieurs rôles, comme celui de réguler des événements cellulaires. Considérant l’implication des lipides dans la biologie et la capacité du MALDI IMS à les analyser, nous avons développé des stratégies analytiques pour la manipulation des échantillons et l’analyse de larges ensembles de données lipidiques. La dégradation des lipides est très importante dans l’industrie alimentaire. De la même façon, les lipides des sections tissulaires risquent de se dégrader. Leurs produits de dégradation peuvent donc introduire des artefacts dans l’analyse IMS ainsi que la perte d’espèces lipidiques pouvant nuire à la précision des mesures d’abondance. Puisque les lipides oxydés sont aussi des médiateurs importants dans le développement de plusieurs maladies, leur réelle préservation devient donc critique. Dans les études multi-institutionnelles où les échantillons sont souvent transportés d’un emplacement à l’autre, des protocoles adaptés et validés, et des mesures de dégradation sont nécessaires. Nos principaux résultats sont les suivants : un accroissement en fonction du temps des phospholipides oxydés et des lysophospholipides dans des conditions ambiantes, une diminution de la présence des lipides ayant des acides gras insaturés et un effet inhibitoire sur ses phénomènes de la conservation des sections au froid sous N2. A température et atmosphère ambiantes, les phospholipides sont oxydés sur une échelle de temps typique d’une préparation IMS normale (~30 minutes). Les phospholipides sont aussi décomposés en lysophospholipides sur une échelle de temps de plusieurs jours. La validation d’une méthode de manipulation d’échantillon est d’autant plus importante lorsqu’il s’agit d’analyser un plus grand nombre d’échantillons. L’athérosclérose est une maladie cardiovasculaire induite par l’accumulation de matériel cellulaire sur la paroi artérielle. Puisque l’athérosclérose est un phénomène en trois dimension (3D), l'IMS 3D en série devient donc utile, d'une part, car elle a la capacité à localiser les molécules sur la longueur totale d’une plaque athéromateuse et, d'autre part, car elle peut identifier des mécanismes moléculaires du développement ou de la rupture des plaques. l'IMS 3D en série fait face à certains défis spécifiques, dont beaucoup se rapportent simplement à la reconstruction en 3D et à l’interprétation de la reconstruction moléculaire en temps réel. En tenant compte de ces objectifs et en utilisant l’IMS des lipides pour l’étude des plaques d’athérosclérose d’une carotide humaine et d’un modèle murin d’athérosclérose, nous avons élaboré des méthodes «open-source» pour la reconstruction des données de l’IMS en 3D. Notre méthodologie fournit un moyen d’obtenir des visualisations de haute qualité et démontre une stratégie pour l’interprétation rapide des données de l’IMS 3D par la segmentation multivariée. L’analyse d’aortes d’un modèle murin a été le point de départ pour le développement des méthodes car ce sont des échantillons mieux contrôlés. En corrélant les données acquises en mode d’ionisation positive et négative, l’IMS en 3D a permis de démontrer une accumulation des phospholipides dans les sinus aortiques. De plus, l’IMS par AgLDI a mis en évidence une localisation différentielle des acides gras libres, du cholestérol, des esters du cholestérol et des triglycérides. La segmentation multivariée des signaux lipidiques suite à l’analyse par IMS d’une carotide humaine démontre une histologie moléculaire corrélée avec le degré de sténose de l’artère. Ces recherches aident à mieux comprendre la complexité biologique de l’athérosclérose et peuvent possiblement prédire le développement de certains cas cliniques. La métastase au foie du cancer colorectal (Colorectal cancer liver metastasis en anglais, CRCLM) est la maladie métastatique du cancer colorectal primaire, un des cancers le plus fréquent au monde. L’évaluation et le pronostic des tumeurs CRCLM sont effectués avec l’histopathologie avec une marge d’erreur. Nous avons utilisé l’IMS des lipides pour identifier les compartiments histologiques du CRCLM et extraire leurs signatures lipidiques. En exploitant ces signatures moléculaires, nous avons pu déterminer un score histopathologique quantitatif et objectif et qui corrèle avec le pronostic. De plus, par la dissection des signatures lipidiques, nous avons identifié des espèces lipidiques individuelles qui sont discriminants des différentes histologies du CRCLM et qui peuvent potentiellement être utilisées comme des biomarqueurs pour la détermination de la réponse à la thérapie. Plus spécifiquement, nous avons trouvé une série de plasmalogènes et sphingolipides qui permettent de distinguer deux différents types de nécrose (infarct-like necrosis et usual necrosis en anglais, ILN et UN, respectivement). L’ILN est associé avec la réponse aux traitements chimiothérapiques, alors que l’UN est associé au fonctionnement normal de la tumeur.

The Role of ‘Edutainment’ in Education for Sustainable Development

The Sustainable Strategies Game (SSG) is being developed as ‘edutainment’ in response to the need to understand sustainable futures and advocate sustainability within workplaces in Higher Education. SSG seeks to both deliver experiential teaching and learning for business sustainability and enhance students’ learning experiences within Worcester Business School. This paper presents findings from action research undertaken to formally investigate two aspects of SSG within edutainment for ESD: firstly, it explores the value students obtain from game playing as an approach to sustainability learning. Secondly, it establishes students’ suggestions for evolutions to SSG, e.g. game design and additional features such as social media interventions or legal challenges, to increase its value as a tool for teaching and learning. Informal feedback following sessions playing SSG suggests games generally generate positive effects on students’ learning. Students highlighted SSG offered an enjoyable alternative approach to learning and could drive changes to sustainability thinking. Introducing such gameplay offers the potential to engage participants in collaborative behaviours and encourage consideration of profitability through strategies which carry less impact on the environment; vital to create a sustainable future. This paper presents qualitative evidence from game players that can enhance SSG as a tool to further improve students’ learning experience and its value as edutainment rather than entertainment within ESD.

Professional Practice in Learning and Development: How to Design and Deliver Plans for the Workplace

Introduction The world is changing! It is volatile, uncertain, complex and ambiguous. As cliché as it may sound the evidence of such dynamism in the external environment is growing. Business-as-usual is more of the exception than the norm. Organizational change is the rule; be it to accommodate and adapt to change, or instigate and lead change. A constantly changing environment is a situation that all organizations have to live with. What makes some organizations however, able to thrive better than others? Many scholars and practitioners believe that this is due to the ability to learn. Therefore, this book on developing Learning and Development (L&D) professionals is timely as it explores and discusses trends and practices that impact organizations, the workforce and L&D professionals. Being able to learn and develop effectively is the cornerstone of motivation as it helps to address people’s need to be competent and to be autonomous (Deci & Ryan, 2002; Loon & Casimir, 2008; Ryan & Deci, 2000). L&D stimulates and empowers people to perform. Organizations that are better at learning at all levels; the individual, group and organizational level, will always have a better chance of surviving and performing. Given the new reality of a dynamic external environment and constant change, L&D professionals now play an even more important role in their organizations than ever before. However, L&D professionals themselves are not immune to the turbulent changes as their practices are also impacted. Therefore, the challenges that L&D professionals face are two-pronged. Firstly, in relation to helping and supporting their organization and its workforce in adapting to the change, whilst, secondly developing themselves effectively and efficiently so that they are able to be one-step ahead of the workforce that they are meant to help develop. These challenges are recognised by the CIPD, as they recently launched their new L&D qualification that has served as an inspiration for this book. L&D plays a crucial role at both strategic (e.g. organizational capability) and operational (e.g. delivery of training) levels. L&D professionals have moved from being reactive (e.g. following up action after performance appraisals) to being more proactive (e.g. shaping capability). L&D is increasingly viewed as a driver for organizational performance. The CIPD (2014) suggest that L&D is increasingly expected to not only take more responsibility but also accountability for building both individual and organizational knowledge and capability, and to nurture an organizational culture that prizes learning and development. This book is for L&D professionals. Nonetheless, it is also suited for those studying Human Resource Development HRD at intermediate level. The term ‘Human Resource Development’ (HRD) is more common in academia, and is largely synonymous with L&D (Stewart & Sambrook, 2012) Stewart (1998) defined HRD as ‘the practice of HRD is constituted by the deliberate, purposive and active interventions in the natural learning process. Such interventions can take many forms, most capable of categorising as education or training or development’ (p. 9). In fact, many parts of this book (e.g. Chapters 5 and 7) are appropriate for anyone who is involved in training and development. This may include a variety of individuals within the L&D community, such as line managers, professional trainers, training solutions vendors, instructional designers, external consultants and mentors (Mayo, 2004). The CIPD (2014) goes further as they argue that the role of L&D is broad and plays a significant role in Organizational Development (OD) and Talent Management (TM), as well as in Human Resource Management (HRM) in general. OD, TM, HRM and L&D are symbiotic in enabling the ‘people management function’ to provide organizations with the capabilities that they need.

Science-based entrepreneur project development process for pre-service science teachers: difficulties faced*

This study used a phenomenological research design to determine the difficulties faced in the science-based entrepreneur project development process for pre-service science teachers.. Qualitative data were obtained through interviews conducted with ten pre-service science teachers. The data were analysed using an inductive thematic analysis. The results indicated that pre-service science teachers have most difficulty ‘making decisions on one of the innovative ideas’ and ‘making predictions about unexpected situations’. They also have difficulties ‘calculating the cost as a result of design or work analysis’, ‘identifying if the idea already existed (similarity analysis)’ and ‘making decisions on the required materials, tools, services’. These results show the need for pre-service science teachers to communicate with other institutions and organisations.

Towards Empowerment: Narrative Study of Counselling Psychology Trainees and How They Make Sense of Their Personal and Professional Development in the Context of Their Past Experiences

The discipline of counselling psychology continues to grow and change in response to social, economic and political pressures. It has been argued that its quest for a coherent and distinct identity, which emphasises the possibility of the coexistence of multiple approaches, creates an inherently uncertain and dilemmatic training environment that may hinder the development of trainees’ professional identities. In order to gain a deeper understanding of the issue at hand, the aim was to explore how final year trainees and newly qualified counselling psychologists constructed and made sense of their emerging professional identities and what experiences, past and present, they drew upon in the context of their training to shape those identities. Applying narrative inquiry to analyse eight open-ended interviews, eight preliminary themes were originally identified in participants’ narratives, which with further refinements lead to stories of struggle and marginalisation, growth and discovery, and power and resilience. Participants’ stories of struggle and marginalisation emerged in reference to early family dynamics and stressful life experiences, which seemed to also foster a strong identification with the counselling psychology profession, while stories of growth and discovery focused on the importance of having supportive figures, who helped to instill a sense of security and create an atmosphere of openness. It was in this learning environment that participants felt it was possible to develop a more resilient, empowered professional self, which allowed them to shed an earlier sense of struggle and vulnerability. However, where more of an emphasis was placed on power and resilience, there seemed to be less room for participants to express other feelings that came into conflict with their preferred sense of professional self. While there seems to be a need for a ‘safer’ climate, in which trainees could voice and acknowledge anxieties, vulnerabilities and limitations, addressing concerns around power and vulnerability that may be contributing to the silencing of particular voices and identities may be equally important if trainees are to develop coherent and distinct counselling psychologist identities.

Design Guidelines and Development Policies for an Adaptable Historic Urban Street: The Case of West Street in Quanzhou, Fujian, China

Thesis (Master's)--University of Washington, 2016-06

Theory development in humanitarian logistics : A framework and three cases

Purpose: There is a need for theory development within the field of humanitarian logistics to understand logistics needs in different stages of a crisis and how to meet these. This paper aims to discuss three dimensions identified in logistics and organization theories and how they relate to three different cases of humanitarian logistics operations - the regional concept of the International Federation of Red Cross Red Crescent Societies, the development and working of the United Nations Joint Logistics Centre and coordination challenges of military logistics in UN mandated peacekeeping operations. The purpose is to build a framework to be used in further studies. Design/methodology/approach: A framework for the study of humanitarian logistics along three dimensions is developed, followed by a discussion of the chosen cases in relation to these dimensions. The framework will be used as basis for the case studies to be undertaken for the purpose of understanding and identification of new questions and needs for other or revised concepts from theory. Findings: The paper shows the relevance of a wide literature to the issues pertinent to humanitarian logistics. There is considerable promise in extant literature on logistics, SCM and coordination, but this needs to be confronted with the particular issues seen in the humanitarian logistics setting to achieve further theory development. Originality/value: The major contribution of the paper lies in its breadth of theoretical perspectives presented and combined in a preliminary theoretical framework. This is applied more specifically in the three case studies described in the paper.

Development of variable displacement oil pump for internal combustion engines

In the last years the need to develop more environmentally friendly and efficient cars as led to the development of several technologies to improve the performance of internal combustion engines, a large part of the innovations are focused in the auxiliary systems of the engine, including, the oil pump, this is an element of great importance in the dynamics of the engine as well a considerable energy consumer. Most solutions for oil pumps to this day are fixed displacement, for medium and high speeds, the pump flow rate is higher than the needs of the engine, this excess flow leads to the need for recirculation of the fluid which represents a waste of energy. Recently, technological advances in this area have led to the creation of variable displacement oil pumps, these have become a 'must have' due to the numerous advantages they bring, although the working principle of vane or piston pumps is relatively well known, the application of this technology for the automotive industry is new and brings new challenges. The focus of this dissertation is to develop a new concept of variable displacement system for automotive oil pumps. The main objective is to obtain a concept that is totally adaptable to existing solutions on the market (engines), both dimensionally as in performance specifications, having at the same time an innovative mechanical system for obtaining variable displacement. The developed design is a vane pump with variable displacement going in line with existing commercial solutions, however, the variation of the eccentricity commonly used to provide an variable displacement delivery is not used, the variable displacement is achieved without varying the eccentricity of the system but with a variation of the length of the pumping chamber. The principle of operation of the pump is different to existing solutions while maintaining the ability to integrate standard parts such as control valves and mechanical safety valves, the pump is compatible with commercial solutions in terms of interfaces for connection between engine systems and pump. A concept prototype of the product was obtained in order to better evaluate the validity of the concept. The developed concept represents an innovation in oil pumps design, being unique in its mechanical system for variable displacement delivery.

DEVELOPMENT OF A DECISION/DEPLOYMENT SUPPORT TOOL FOR VARIABLE SPEED LIMITS IN RECURRENT CONGESTION

Traffic demand increases are pushing aging ground transportation infrastructures to their theoretical capacity. The result of this demand is traffic bottlenecks that are a major cause of delay on urban freeways. In addition, the queues associated with those bottlenecks increase the probability of a crash while adversely affecting environmental measures such as emissions and fuel consumption. With limited resources available for network expansion, traffic professionals have developed active traffic management systems (ATMS) in an attempt to mitigate the negative consequences of traffic bottlenecks. Among these ATMS strategies, variable speed limits (VSL) and ramp metering (RM) have been gaining international interests for their potential to improve safety, mobility, and environmental measures at freeway bottlenecks. Though previous studies have shown the tremendous potential of variable speed limit (VSL) and VSL paired with ramp metering (VSLRM) control, little guidance has been developed to assist decision makers in the planning phase of a congestion mitigation project that is considering VSL or VSLRM control. To address this need, this study has developed a comprehensive decision/deployment support tool for the application of VSL and VSLRM control in recurrently congested environments. The decision tool will assist practitioners in deciding the most appropriate control strategy at a candidate site, which candidate sites have the most potential to benefit from the suggested control strategy, and how to most effectively design the field deployment of the suggested control strategy at each implementation site. To do so, the tool is comprised of three key modules, (1) Decision Module, (2) Benefits Module, and (3) Deployment Guidelines Module. Each module uses commonly known traffic flow and geometric parameters as inputs to statistical models and empirically based procedures to provide guidance on the application of VSL and VSLRM at each candidate site. These models and procedures were developed from the outputs of simulated experiments, calibrated with field data. To demonstrate the application of the tool, a list of real-world candidate sites were selected from the Maryland State Highway Administration Mobility Report. Here, field data from each candidate site was input into the tool to illustrate the step-by-step process required for efficient planning of VSL or VSLRM control. The output of the tool includes the suggested control system at each site, a ranking of the sites based on the expected benefit-to-cost ratio, and guidelines on how to deploy the VSL signs, ramp meters, and detectors at the deployment site(s). This research has the potential to assist traffic engineers in the planning of VSL and VSLRM control, thus enhancing the procedure for allocating limited resources for mobility and safety improvements on highways plagued by recurrent congestion.

Towards guidelines for curriculum design and development in English language teacher education in Cambodian higher education institutions

In the current Cambodian higher education sector, there is little regulation of standards in curriculum design of undergraduate degrees in English language teacher education. The researcher, in the course of his professional work in the Curriculum and Policy Office at the Department of Higher Education, has seen evidence that most universities tend to copy their curriculum from one source, the curriculum of the Institute of Foreign Languages, the Royal University of Phnom Penh. Their programs fail to impose any entry standards, accepting students who pass the high school exam without any entrance examination. It is possible for a student to enter university with satisfactory scores in all subjects but English. Therefore, not many graduates are able to fulfil the professional requirements of the roles they are supposed to take. Neau (2010) claims that many Cambodian EFL teachers do not reach a high performance standard due to their low English language proficiency and poor background in teacher education. The main purpose of this study is to establish key guidelines for developing curricula for English language teacher education for all the universities across the country. It examines the content of the Bachelor‘s degree of Education in Teaching English as a Foreign Language (B Ed in TEFL) and Bachelor‘s degree of Arts in Teaching English to Speakers of Other Languages (BA in TESOL) curricula adopted in Cambodian universities on the basis of criteria proposed in current curriculum research. It also investigates the perspectives of Cambodian EFL teachers on the areas of knowledge and skill they need in order to perform their English teaching duties in Cambodia today. The areas of knowledge and skill offered in the current curricula at Cambodian higher education institutions (HEIs), the framework of the knowledge base for EFL teacher education and general higher education, and the areas of knowledge and skill Cambodian EFL teachers perceive to be important, are compared so as to identify any gaps in the current English language teacher education curricula in the Cambodian HEIs. The existence of gaps show what domains of knowledge and skill need to be included in the English language teacher education curricula at Cambodian HEIs. These domains are those identified by previous curriculum researchers in both general and English language teacher education at tertiary level. Therefore, the present study provides useful insights into the importance of including appropriate content in English language teacher education curricula. Mixed methods are employed in this study. The course syllabi and the descriptions within the curricula in five Cambodian HEIs are analysed qualitatively based on the framework of knowledge and skills for EFL teachers, which is formed by looking at the knowledge base for second language teachers suggested by the methodologists and curriculum specialists whose work is elaborated on the review of literature. A quantitative method is applied to analyse the perspectives of 120 Cambodian EFL teachers on areas of knowledge and skills they should possess. The fieldwork was conducted between June and August, 2014. The analysis reveals that the following areas are included in the curricula at the five universities: communication skills, general knowledge, knowledge of teaching theories, teaching skills, pedagogical reasoning and decision making skills, subject matter knowledge, contextual knowledge, cognitive abilities, and knowledge of social issues. Additionally, research skills are included in three curricula while society and community involvement is in only one. Further, information and communication technology, which is outlined in the Education Strategies Plan (2006-2010), forms part of four curricula while leadership skills form part of two. This study demonstrates ultimately that most domains that are directly and indirectly related to language teaching competence are not sufficiently represented in the current curricula. On the basis of its findings, the study concludes with a set of guidelines that should inform the design and development of TESOL and TEFL curricula in Cambodia.

Status of the beach management units development in Uganda

Fisheries plays a significant and important part in the economy of the country contributing to foreign exchange, food security and employment creation. Lake Victoria contributes over 50% of the total annual fish catch. The purpose of fisheries management is to ensure conservation, protection, proper use, economic efficiency and equitable distribution of the fisheries resources both for the present and future generations through sustainable utilization. The earliest fisheries were mainly at the subsistence level. Fishing gear consisted of locally made basket traps, hooks and seine nets of papyrus. Fishing effort begun to increase with the introduction of more efficient flax gillnets in 1905. Fisheries management in Uganda started in 1914. Before then, the fishery was under some form of traditional management based on the do and don'ts. History shows that the Baganda had strong spiritual beliefs in respect of "god Mukasa" (god of the Lake) and these indirectly contributed to sustainable management of the lake. If a fisherman neglected to comply witt'l any of the ceremonies related to fishing he was expected to encounter a bad omen (Rev. Roscoe, 1965) However, with the introduction of the nylon gill nets, which could catch more fish, traditional management regime broke down. By 1955 the indigenous fish species like Oreochromis variabilis and Oreochromis esculentus had greatly declined in catches. Decline in catches led to introduction of poor fishing methods because of competition for fish. Government in an attempt to regulate the fishing irldustry enacted the first Fisheries Ordinance in 1951 and recruited Fisheries Officers to enforce them. The government put in place minimum net mesh-sizes and Fisheries Officers arrested fishermen without explaining the reason. This led to continued poor fishing practices. The development of government centred management systems led to increased alienation of resource users and to wilful disregard of specific regulations. The realisation of the problems faced by the central management system led to the recognition that user groups need to be actively involved in fisheries management if the systems are to be consistent with sustainable fisheries and be legitimate. Community participation in fisheries management under the Comanagement approach has been adopted in Lake Victoria including other water bodies.

The role of microRNA 21 in the development of in-stent restenosis

Coronary heart disease is a major cause of morbidity and mortality worldwide. Percutaneous coronary intervention (PCI) has become the most widely used method of coronary artery revascularisation. The use of stents to hold open atherosclerosis induced arterial narrowing has significantly reduced elastic recoil and acute vessel occlusion following balloon angioplasty. However, bare metal stents have been associated with in-stent restenosis attributed to vascular smooth muscle cell (VSMC) hyperplasia and excessive neointimal formation. The resultant luminal renarrowing may manifest clinically with the return of symptoms such as chest pain or shortness of breath. The development of drug eluting stents has significantly reduced the incidence of in-stent restenosis (ISR). Unfortunately the antiproliferative medications used not only inhibit VSMC proliferation but also re-endothelialisation of the stented vessel. In addition, the drug impregnated polymer coating has been associated with a chronic inflammatory response within the vessel wall predisposing patients to stent thrombosis. Thus the identification of novel therapies which promote vessel healing without excessive proliferative or inflammatory response may improve long term outcome and reduce the need for repeated revascularisation. MicroRNAs (miRs) are short (18-25 nucleotide) non-coding RNAs acting to regulate gene expression. By binding to the 3’untranslated region of mRNA they act to fine tune gene expression either by mRNA degradation or translational repression. Originally identified in coordinating tissue development microRNAs have also been shown to play important roles coordinating the inflammatory response and in numerous cardiovascular diseases. MiR-21 has been identified in human atherosclerotic plaques, arteriosclerosis obliterans and abdominal aortic aneurysms. In addition, its up regulation has been documented in preclinical models of vascular injury. This study sought to identify the role of miR-21 in the development of ISR. Utilising a small animal model of stenting and in vitro techniques, we sought to investigate its influence upon VSMC and immune cell response following stenting. 19 The refinement of a murine stenting model within the Baker laboratory and the electrochemical dissolution of the metal stent from within harvested vascular tissues significantly improved the ability to perform detailed histological analysis. In addition, identification of miRNAs using in situ hybridisation was achieved for the first time within stented tissue. Neointimal formation and ISR was significantly reduced in mice in which miR-21 had been genetically deleted. In addition, neointimal composition was found to be altered in miR-21 KO mice with reductions in VSMC and elastin content demonstrated. Importantly, no difference in re-endothelialisation was observed. In vitro analysis demonstrated that VSMCs from miR-21 KO mice had both reduced proliferative and migratory capacity following platelet derived growth factor stimulation. Molecular analysis revealed that these differences may, at least in part, be due to de-repression of programmed cell death 4 (PDCD4). PDCD4 is a known miR-21 target within VSMCs implicated in the suppression of proliferation and promotion of apoptosis. Unfortunately, initial attempts at antimiR mediated knockdown of miR-21 in vivo, failed to produce a similar change in the suppression of ISR. Furthermore, a significant alteration in macrophage polarisation state within the neointima of miR-21 WT and KO mice was noted. Immunohistochemical staining revealed a preponderance of anti-inflammatory M2 macrophages in KO mice. Analysis of bone marrow derived macrophages from miR-21 KO mice demonstrated an increased level of the peroxisome proliferation activating receptor-γ (PPARγ) which facilitates M2 polarisation. Importantly, significant alterations in numerous pro-inflammatory cytokines, which also have mitogenic effects, were also found following genetic deletion of miR-21. In Summary, this is the first study to look at miRs in the development of ISR. MiR-21 plays an important role in the development of ISR by influencing the proliferative response of VSMCs and modulating the immune response following stent deployment. Further attempts to modulate miR-21 expression following PCI may reduce ISR and the need for repeat revascularisation while also reducing the risk of stent thrombosis.

Investigating the role of N-WASP in the development of colorectal cancer

Colorectal cancer (CRC) is the second most common cancer in Europe, with the second highest mortality rate. Although prognosis is improving, survival rates remain poor for those presenting with the most advanced stages of the disease. There is therefore a need for improved early diagnosis and thus a greater understanding of the early stages of the development of colorectal tumours is desirable. Additionally, as most deaths in colorectal cancer are due to advanced metastatic disease, it is of great interest to explore any potential mechanisms by which metastatic disease can be inhibited. N-WASP is a ubiquitously expressed protein with multiple intracellular roles including actin regulation and maintaining stability of epithelial cell-cell junctions. Through its role as an actin regulator, it has been implicated in the processes of invasion and metastasis of multiple cancer types. Its role in the development and progression of colorectal cancer however has not been fully explored. This thesis will present a series of in vitro and in vivo studies that were carried out with the aim of answering the following questions: • Does N-Wasp have a role in normal intestinal homeostasis? • Does N-Wasp knockout affect the development of tumours in a mouse model of intestinal tumourigenesis? • Does N-Wasp knockout affect the invasive properties of intestinal cancer in vitro? • Does N-WASP correlate with prognosis or other indicators in human colorectal cancer TMAs? Findings from the in vivo experiments, using an inducible, gut-specific knockout model, have uncovered potential roles for N-Wasp in regulating differentiation and migration of intestinal epithelial cells. Although it had no effect in short term models of intestinal hyperproliferation, N-Wasp knockout increased tumour burden and decreased survival in an established in vivo model of intestinal tumourigenesis, in which there is heterozygous loss of Apc (Apcfl/+). No effect was seen on tumour development or survival when additional N-WASP knockout was introduced into a more rapid model, with heterozygous loss of Apc and mutation of Kras (Apcfl/+ KrasG12D/+). N-WASP expression in human colorectal cancer was assessed using immunohistochemical staining of two tissue microarrays. Low levels of N-WASP expression were found to be associated with presence of MMR deficiency. There was no statistically significant difference in overall or cancer specific survival based on N-WASP expression. Collectively, the data presented here suggest a previously unreported role for N-WASP in regulation of intestinal epithelial differentiation and indicate that it may act as a tumour suppressor against development of benign precursor lesions of colorectal cancer. Further research is warranted to delineate the mechanisms underlying these processes.