An Open Research University

This is the final report of the Open University’s RCUK-funded Public Engagement with Research Catalyst, ‘An open research university’, a project designed to create the conditions in which engaged research can flourish. The report describes an evidence-based strategy designed to embed engaged research within the University’s strategic planning for research and the operational practices of researchers. This programme of organisational change was informed by action research, working collaboratively with researchers at all levels across the institution to identify and implement strategies that work for them and the stakeholders, user communities and members of the public that engage with their research. Through a combination of surveys, interviews and interventions, we identified a number of challenges and proposed solutions to address them. For example, we found that researchers have a relatively narrow view of engaged research and the communities with which they interact and very few researchers strategically evaluate their engaged research activities. The report documents some of the interventions we have introduced with the aim of broadening and deepening future researcher engagement, including a definition of engaged research and revised promotion criteria that include knowledge exchange profiles. In conclusion, we argue that there is still a battle to be won for open and engaged research. For a culture of engaged research to be sustainable in the medium to long-term requires ongoing recognition and acceptance of its progressive value(s) by researchers, universities, funders and ultimately, policy-makers

Ferumoxytol-enhanced magnetic resonance imaging in acute myocarditis

Objectives Ultrasmall superparamagnetic particles of iron oxide (USPIO)-enhanced MRI can detect tissue-resident macrophage activity and identify cellular inflammation within tissues. We hypothesised that USPIO-enhanced MRI would provide a non-invasive imaging technique that would improve the diagnosis and management of patients with acute myocarditis. Methods Ten volunteers and 14 patients with suspected acute myocarditis underwent T2, T2* and late gadolinium enhancement (LGE) 3T MRI, with further T2* imaging at 24 hours after USPIO (ferumoxytol, 4 mg/kg) infusion, at baseline and 3 months. Myocardial oedema and USPIO enhancement were determined within areas of LGE as well as throughout the myocardium. Results Myocarditis was confirmed in nine of the 14 suspected cases of myocarditis. There was greater myocardial oedema in regions of LGE in patients with myocarditis when compared with healthy volunteer myocardium (T2 value, 57.1±5.3 vs 46.7±1.6 ms, p0.05). Imaging after 3 months in patients with myocarditis revealed a reduction in volume of LGE, a reduction in oedema measures within regions displaying LGE and improvement in ejection fraction (mean −19.7 mL, 95% CI (−0.5 to −40.0)), −5.8 ms (−0.9 to −10.7) and +6% (0.5% to 11.5%), respectively, p<0.05 for all). Conclusion In patients with acute myocarditis, USPIO-enhanced MRI does not provide additional clinically relevant information to LGE and T2 mapping MRI. This suggests that tissue-resident macrophages do not provide a substantial contribution to the myocardial inflammation in this condition. Clinical trial registration NCT02319278; Results

Characterisation of microRNAs in human stem cells

In collaboration with David Baulcombe and Attila Molnar we have generated microRNA libraries for human embryonic stem cells (hESCs) before and after differentiation along the neuronal lineage and also from human mesenchymal stem cells (hMSCs). Both cell types are of medical importance and understanding how their proliferation and differentiation is regulated by microRNAs is also of scientific interest. The hMSC library was sequenced by 454 technology and the two subsequent hESC libraries by Solexa sequencing. Approximately a quarter of all currently known microRNAs were identified between the libraries, in addition to 3 novel microRNAs and 25 annotated piRNAs. For the hESC libraries, we verified the presence of embryonic specific microRNAs (miR-302 family) and neuronal specific microRNAs (miR-9/miR-9*), and demonstrated that expression of these miRNAs is regulated at the transcriptional level. Additionally, promoter assessments of miR-9 transcription revealed that multiple upstream regions may be important in neuronal specific expression. Almost half of all known human microRNAs are located within the introns of host genes. We used microarrays to analyse host gene expression and found that there was little correlation with microRNA expression, indicating that many microRNAs are not regulated at the transcriptional level by their host promoter. Furthermore, the expression of microRNAs from the same cluster, and also from the same hairpin precursor, did not always correlate when compared between the stem cell libraries. Taken together, this data indicates that microRNAs are regulated at a variety of levels both pre- and post-transcriptionally. Many microRNA isomers were also detected that differed in expression between human cell types, and upon differentiation of the hMSCs through the osteoblastic lineage. Interestingly, microRNAs and some of their isomers showed different affinities for Argonaute proteins in pulldown assays. We also profiled mRNAs that were immunoprecipitated with Argonaute in order to identify miRNA targetsEThOS - Electronic Theses Online ServiceBBSRC, Institute of Obstetrics and Gynaecology Trust.GBUnited Kingdo

A study to improve the differentiation of human embryonic stem cells to functional hepatocytes

Human embryonic stem cells (hESCs) possess 2 unique properties (1) pluripotency and (2) self-renewal, and therefore, hold great promise for biomedical application and regenerative medicine. In vitro differentiation of hESCs is a vital tool to generate unlimited human hepatocytes. To date, several multi-step protocols have been established to generate hepatocyte-like cells from undifferentiated hESCs via definitive endoderm (DE) formation. However, hESC derived-hepatocytes in these systems exhibit some immature characteristics, thus it remains a challenge on how to further improve hepatic differentiation. In addition, the molecular mechanisms regulating the differentiation are still ambiguous, making in vitro differentiation a difficult task. The ultimate aim of this project is to improve the differentiation of hESCs to functional hepatocytes. In this thesis, the work includes two main parts: (1) modulating signalling pathways to explore molecular mechanisms controlling DE differentiation; (2) developing a 3-dimensional (3D) culture system to improve the functionality of hESC-derived hepatocytes. DE formation is a critical step for the production of hepatocytes. In the first part of this thesis, I showed that suppression of PI3K signalling using the LY 294002 inhibitor (LY) during hESC differentiation significantly improves Activin A (AA)-induced DE generation, which subsequently augments hepatocyte production. Further mechanistic interrogation of this phenomenon has revealed that dual treatment of hESCs with AALY enhances the Activin downstream signalling, Smad2/3 phosphorylation and their nuclear translocation with Smad4. Furthermore, dual treatment with AALY also affects the disruption of β-catenin/E-cadherin complexes, which cooperatively contribute to distinctive morphological changes that may signify the occurrence of EMT and hence improved specification of DE. These findings suggest that suppression of PI3K/Akt modulates both Nodal/Activin and β-catenin pathways, the two most important signalling involved in mesendoderm and DE cell fate specification, therefore improved DE differentiation of hESCs. Liver development in vivo is regulated by cell–cell contacts in a 3D environment and the absences of this may account for, at least partly, some of the immature features of hESC-derived hepatocytes. In the second part of my thesis, based on initial encouraging results obtained from HepG2 cells, I applied alginate based 3D culture system to hESCs after they are differentiated into DE cells and optimised culture conditions. The results confirmed that 3D culture microenvironment enhanced hepatic differentiation and functionality of hESC-derived hepatocytes in compared to the monolayer format. Collectively, this study has demonstrated a significant cornerstone in the strategies to improve hepatic differentiation of hESCs by addressing the molecular signalling and micro-niche cues that govern hepatocyte lineage commitment. Hence, this will pave the way for the use of these hepatocytes in future regenerative therapies and biomedical applications.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Muscular dystrophy cell therapy : an in utero approach using human fetal mesenchymal stem cells

Duchenne muscular dystrophy (DMD) is the most prevalent genetic neuromuscular disorder and affects 1 in 3,500 live male births. Lack of the protein dystrophin in muscle fibres causes permanent muscle damage, is lethal and despite various potential therapeutic strategies aimed at restoring dystrophin expression, has no cure. As DMD affects all skeletal muscles as well as the heart, a systemic treatment would be necessary and in utero stem cell transplantation is a promising way of achieving this. The identification of human fetal mesenchymal stem cells (hfMSC) in early gestation fetal blood offers the prospect of allogeneic or autologous cell therapy, while intrauterine administration would capitalise on ontological opportunities unique to the developing fetus. The aim of the study was to improve hfMSC engraftment and contribution to skeletal muscle fibres following intrauterine transplantation (IUT) in a mouse model of DMD. My project demonstrated that hfMSCs are easily isolated and expandable with the ability to undergo myogenesis in vitro. HfMSCs differentiated into mature myotubes following exposure to galectin-1 conditioned medium, while galectin-1 transduced hfMSCs showed significantly higher expression of myogenic markers compared to non-transduced hfMSCs. Co-culture experiments provided an in vitro model to explore the underlying mechanism for muscle differentiation of hfMSCs following IUT. HfMSCs were able to form chimeric myotubes by fusing with myoblasts isolated from E15 mouse embryos, evidence that they should be able to fuse with developing muscle fibres in vivo. Engraftment and differentiation into muscle fibres of hfMSCs injected intra-peritoneally into E15 mouse embryos in vivo was enhanced by using immunodeficient dystrophic host mice, postnatal muscle injury and additional neonatal hfMSC transplantation following IUT. In conclusion, my thesis supports the use of hfMSC as an attractive source for cell therapy and provides the background for further studies to optimise their engraftment and differentiation to underpin future clinical applications.EThOS - Electronic Theses Online ServiceWellbeing of Women and Institute of Obstetrics and Gynaecology TrustGBUnited Kingdo

Switching on kinases: oncogenic activation of BRAF and the PDGFR family

The cytoplasmic serine/threonine kinase BRAF and receptor tyrosine kinases of the platelet-derived growth factor receptor (PDGFR) family are frequently activated in cancer by mutations of an equivalent amino acid. Structural studies have provided important insights into why these very different kinases share similar oncogenic hot spots and why the PDGFR juxtamembrane region is also a frequent oncogenic target. This research has implications for other kinases that are mutated in human tumours and for the treatment of cancer using kinase inhibitors

Maintaining the strategic edge: the defence of Australia in 2015

The recent and continuing changes in Southeast Asia - the economic calamity in 1997-98, the overthrow of President Soeharto's New Order and the tenuous establishment of democracy, and the horrific circumstances of East Timor's independence - have disturbed Australia's security situation more seriously than anything since the 1960s, when Australia was at war (albeit covertly) with Indonesia in Borneo and had a task force in Vietnam. The rate of technological change is also unprecedented, especially in the area of information technology (IT) and its manifold applications, promising a revolution in military affairs (RMA), some aspects of which are very attractive for Australian defence planning. At the same time, the Australian Defence Force (ADF) faces the imminent prospect of 'block obsolescence' - when major platforms such as the F/A-18 Hornet multi-role fighter aircraft, the F-111 strike fighters, the P-3C Orion long-range maritime patrol aircraft, and all of the navy's surface combatants, will need to be replaced (or their tasks foregone). Addressing these issues will require the development of a sound appreciation of Australia's security environment, and of clear and coherent strategic guidance for defence force planning. The purpose of this volume is to assist and inform these processes

Cryptic splice sites and split genes

We describe a new program called cryptic splice finder (CSF) that can reliably identify cryptic splice sites (css), so providing a useful tool to help investigate splicing mutations in genetic disease. We report that many css are not entirely dormant and are often already active at low levels in normal genes prior to their enhancement in genetic disease. We also report a fascinating correlation between the positions of css and introns, whereby css within the exons of one species frequently match the exact position of introns in equivalent genes from another species. These results strongly indicate that many introns were inserted into css during evolution and they also imply that the splicing information that lies outside some introns can be independently recognized by the splicing machinery and was in place prior to intron insertion. This indicates that non-intronic splicing information had a key role in shaping the split structure of eukaryote genes. <br/

Get MediaSmart®: A critical discourse analysis of controversy around advertising to children in the UK

In response to calls for increased regulation of advertising to children (occasioned by concerns over childhood obesity levels) a group of UK advertisers targeting young people have sought to demonstrate social responsibility by providing media literacy education resources for children aged six to eleven through the MediaSmart® initiative. This article draws on Critical Discourse Analysis (Fairclough 2001) to analyse a selection of publicly available accounts of the 2002 launch and operation of MediaSmart® in order to explore how alternative discursive representations of MediaSmart® construct children and advertising in relation to one another, and how these constructions work to further the social practices of which the discourses in question are part. The analysis concludes that the competing discourses have a stake in the problem of advertising to children remaining open-ended, but suggests that the possibilities of its resolution lie in (a) the incorporation of children's own perspectives in controversy conducted on their behalf by adults, and (b) conceptions of media literacy which are more active and age-inclusive than those evident in the discourses currently available