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Results

Defining the Fc receptor-mediated trafficking of IgG-antigen complexes in macrophages 05 Apr 2017

Macrophages represent the most abundant antigen presenting cells in inflammatory infiltrates and tumors. An important immunoregulatory function of macrophages is to present antigen to cognate T cells. Antibody (IgG)-antigen complexes are internalized into macrophages through potential interactions with multiple different Fc receptors and are subsequently delivered to subcellular compartments where peptides derived from the proteolyzed antigen can be loaded onto MHC molecules. However, the multitude of possible Fc receptor interactions with the antibody-antigen complexes influences the spatial and temporal behavior of antigen and consequent peptide-MHC repertoire through a complex, and poorly understood, network of subcellular trafficking pathways. We have developed advanced microscopy tools to enable the analysis of three-dimensional, dynamic cellular processes at unprecedented levels of spatiotemporal resolution. We will combine these approaches with antibody engineering and analyses in mouse models of cancer and autoimmunity to elucidate how the combination of antigen delivery vehicle, microenvironmental factors and macrophage phenotype interplay to result in specific antigen presentation outcomes. The two long term goals of these studies are: one, to develop novel mechanistic insights into the fundamental cell biological processes determining the repertoire of presented antigens; two, to define how to deliver antigen to macrophages to orchestrate a predictable immune response.

Amount: £2,700,000
Funder: The Wellcome Trust
Recipient: University of Southampton

Novel Role of a Protein Phosphatase in Chromosome Segregation 05 Sep 2017

Kinases and their antagonistic phosphatases both provide a key regulatory mechanism controlling many cellular events. Yet our molecular understanding of phosphatases lags behind. Recently we described an interaction between Protein Phosphatase 4 (PP4) and Drosophila centromeres, which when disrupted affects the mitotic centromere integrity and spindle assembly checkpoint activity. The details of this pathway remain to be elucidated. Nor is it known how this finding relates to PP4's de-regulation in many cancers. I will therefore turn to human cell line model to address these questions. Towards determining human PP4’s mitotic functions, CRISPR/Cas9 gene editing will be used to generate cell lines expressing endogenous PP4 fused with the AID tag allowing for inducible and rapid degradation of the phosphatase. This approach will facilitate PP4 removal specifically in mitotic cells, the effects of which will be characterised by cell imaging. Parallel studies, including proteomics, will deliver precise information on the human PP4 interaction network and behaviour during mitosis. Together these data will reveal the affected processes and provide a blueprint for subsequent detailed investigations that identify the involved pathways, PP4-specific substrates and their phospho-regulatory sites. The results of this study will be later placed within a larger investigatory framework dissecting chromosome segregation.

Amount: £100,000
Funder: The Wellcome Trust
Recipient: University of Southampton

What are the molecular mechanisms regulating neuronal sensitivity to BDNF? 31 May 2018

Neurotrophins and their receptors play a key role in the maintenance and remodelling of the nervous system, and defects in trophic signalling are associated with mental illness as well as neurodegeneration. To date, the cellular mechanisms regulating neurotrophin receptor bioavailability are not fully understood. Here we focus on BDNF and its receptor, TrkB, which are widely expressed throughout the central nervous system. This project will investigate the role of two novel potential TrkB regulators that recently emerged from our screens. Candidates will be expressed in HEK-TrkB cells and primary cortical mouse neurons, and (i) TrkB receptor expression levels and (ii) downstream signalling events will be investigated using both Western blotting and immunofluorescence techniques. These experiments will address whether the candidates modulate TrkB availability and activity. A better understanding of the cellular mechanisms underlying TrkB regulation may highlight potential therapeutic targets to counteract TrkB degradation and restore sensitivity of vulnerable cells to the beneficial actions of BDNF.

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Southampton

The role of interferon response genes in an experimental model of Alzheimer's Disease 31 May 2018

Emerging evidence suggest that systemic infection can accelerate the onset and/or progression of cognitive decline in Alzheimer’s disease, but the underlying biological mechanism remain unknown. Exposure of AD mouse models to a real systemic infection results in exaggerated neuroinflammation and AD-like pathology and preliminary microarray analysis suggest this may be due to increased levels of type I or II interferon and/or it’s down-stream signalling pathways. Interferons are produced in response to viral and bacterial infections and well known to be associated with mood changes and depression. In this project we will test if these pro-inflammatory cytokines also play a role in progressing AD-like pathology. Brain tissue from AD mice exposed to a systemic bacterial infection has already been collected, with a control group treated with saline. The expression levels of interferons and their signalling pathways will be explored in this archived tissue and in primary neuronal cultures expressing human Tau. The in vitro cultures will allow us to test if exposure to interferon promotes spreading of tau pathology and if inhibition of the signalling pathways prevents progression of AD-like pathology. This could later be applied to in vivo AD disease models, taking into consideration behavioural changes.

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Southampton

Using a Drosophila model to determine how neuronal clock circuits couple visual inputs to sleep/wake rhythms 31 May 2018

An internal daily timekeeper, known as the circadian clock dictates a 24-hour cycle in behaviour and physiology. In organisms as diverse humans and the genetic model Drosophila melanogaster light perceived by the eyes signals to molecular clock cells in the brain to set their phase. A regular environmental light/dark cycle helps preserve synchrony among different bodily functions, while out-of-phase or abnormal patterns of light exposure can be disruptive to daily timekeeping and result in negative outcomes for both physical mental health and well-being. Given the similarity between the molecular clock circuits of the fruit fly Drosophila and those of mammals, genetic and molecular studies in Drosophila are likely to provide important insight in how visual organs contribute to the control of daily rhythms. To avoid the impact of the invertebrate-specific blue light photoreceptor CRYPTOCHROME, experiments in this proposal will make use of visual organ-detected red light to map light input pathways connecting the Drosophila visual organs to sleep/wake rhythms. In particular, histamine, a known signalling intermediate between the compound eyes and clock neurons, will be examined for its role in synchronizing cellular clocks and regulating intercellular clock signals .

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Southampton

Understanding the formation of Pseudomonas aeruginosa biofilms in urinary catheters 31 May 2018

This proposal uses advanced microscopy and molecular techniques to examine the role of alginate in the attachment of Pseudomonas aeruginosa, and subsequent biofilm formation, on urinary catheters. There is an urgent clinical need for improved catheter management and the development of anti-biofilm materials, however approaches to date have failed and a lack of understanding of biofilm development may contribute to this. In this work, we intend using a simple laboratory model system to generate biofilms on urinary catheters, which can be tracked directly over time using episcopic differential interference contrast microscopy. Transposon mutants (obtained from the PA Two Allele Library) will be used and compared to the type strain, PAO1. In addition, Gibson Assembly will be used to generate knockout mutants for the same genes. The biofilm forming capability of each mutant and type (transposon vs knockout) will be assessed and compared. Transposon mutants have the potential to still produce peptides, with unknown form and function. Such peptides could affect biofilm development. In contrast, knockout mutants remove the target gene sequence. This project will provide information on general biofilm development, the role of alginate, and also indicate whether knockout mutants should be used for understanding the role of specific genes.

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Southampton

The impact of systemic inflammation on the brain. 27 Apr 2017

Brain tissue from adult mice has already been collected, with a control group treated with saline and another treated with an attenuated strain of Salmonella typhimurium, triggering a systemic immune response including production of pro-inflammatory mediators that communicate with the brain triggering neuroinflammation and neuronal dysfunction. Selected groups of mice have been treated with two different inhibitors that block the activity of the cytokine IL-1beta, in order to investigate if this pro-inflammatory cytokine plays a direct role in neuroinflammation during systemic infection. In this project we will compare a small molecule with a biologic, both targeting cytokines, but via different mechanisms. The brain tissue will be examined using antibodies to stain activation markers of microglial cells (CD11b, FcRI, MHCII) and activation markers of cerebral endothelial cells (VCAM, ICAM, MHCII). Synaptic activation markers, from both the pre- and post-synaptic membranes, will be investigated by isolated mRNA from hippocampal enriched tissue. This investigation into phenotypic changes in microglial, cerebral endothelial cells and BBB function will be used to determine if the inhibition of IL-1beta is beneficial in reducing neuroinflammation as a result of systemic infection. This could later be applied to AD models, taking into consideration behavioural changes.

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Southampton

Investigating bacterial transporters that convey antibiotic resistance 27 Apr 2017

The rise of antimicrobial resistance (AMR) threatens many of the major advances in modern healthcare as the treatments rely on the implicit ability of the antibiotic to allow the body to heal from a serious infection. Antibiotic efflux transport is central to the development of AMR. At the single cell level, it is becoming apparent that the use of efflux pumps is the first line of defence against an antibiotic. These pumps decrease the intracellular level of antibiotic while the cell activates the various other levels of protection. This frontline of defence involves a coordinated network of efflux transporters. In the future, inhibition of this efflux transporter network, as a target for novel antibiotic therapy, will require the isolation and then biochemical/biophysical characterisation of each pump against all known and new antibiotics. This depth of knowledge is required so that we can fully understand and tackle the mechanisms of developing antimicrobial resistance. The key goals of this project are to over-express, IMAC purify a hexahistidine-tagged versions of a number of the efflux transporter and then characterise ligand binding against a set of clinically relevant antibiotics. The method used to measure ligand binding is thermophoresis (Nonotemper, Monolith).

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Southampton

Targeting human Langerhans cells to induce long-lasting tolerance in allergy. 21 Oct 2015

Langerhans cells (LCs) orchestrate cutaneous immune responses, inducing immune activation or tolerance to epicutaneously encountered antigens. Here I hypothesise that identification and targeting of the molecular switches controlling antigen presentation in human LCs will enable the induction of efficient and long lasting tolerance in allergy. Applying the systems immunology approach I developed, I aim to compare whole transcriptomes of LCs isolated from clinical samples from paediatric and adul t patients, exposed or not to an allergen, and ex vivo culture of epidermal sheets subjected to contrasting treatments. Computational analysis of molecular networks, paired with silencing of identified candidate targets, and functional validation of immunological outcomes will allow me to dissect the mechanisms of tolerance induction in human skin. I aim to address the following questions: 1) Is LC ability to induce tolerance impaired in atopic skin? 2) How is the allergen presentation re gulated in human LCs during allergen exposure? 3) How can we target the transcriptional networks in human LCs in order to induce long-lasting allergen tolerance? The outlined investigations will lead to a comprehensive understanding of allergen presentation by human LCs, and to the identification of novel therapeutic targets for induction of long-lasting tolerance in allergy.

Amount: £820,382
Funder: The Wellcome Trust
Recipient: University of Southampton

Developing an Ethical Framework for the Implementation of Public Health Regulation 31 May 2016

This project entails a collaboration between the applicant, who has expertise in public health ethics and law, scholars at the University of Western Australia with expertise in law and history, and the Chief Health Officer and Assistant Director General Public Health, Department of Health, Western Australia. It will analyse the historical and regulatory significance of Western Australia’s Public Health Bill (due soon to be enacted), and develop an ethical framework to underpin public health activity under the new law. The key goals are to: Improve understanding, and develop standards for good practice in public health Identify the need and challenges for implementing flexible, principle-led decision making, and will lay the groundwork for an implementation strategy founded in bioethics Establish a platform on which to build the next phases of research, directed at facilitating implementation. Produce generalisable knowledge and insights regarding the new wave of public health laws, which aim to replace outdated understandings of public health with policies that recognise and respond to the social determinants of health, and 21st century public health challenges These goals will be achieved through direct engagement with policy-actors, the generation of academic articles, and the galvanisation of collaborative research links.

Amount: £4,170
Funder: The Wellcome Trust
Recipient: University of Southampton

High resolution global mapping of women of childbearing age, pregnancies and births for Zika burden estimation. 30 Sep 2016

Evidence is mounting on the link between Zika virus infections and birth defects, making an understanding oflikely numbers of pregnancies and births affected a high priority. Assessing the likely demands on healthservices, needs for future vaccines and estimating the current and future burden of the virus requiresaccurate estimates of the numbers at risk. It is clear that Zika transmission risk is spatially heterogenous, andmodelling efforts have recently mapped areas of suitability at high spatial resolution (Messina et al, Perkinset al, Samy et al), and will continue to be refined as more data become available. However, many of theareas predicted as suitable are unpopulated, while others are densely populated. Moreover, the numbers ofbirths and pregnancies can vary substantially within and between countries with significant spatial andtemporal variations in age-specific fertility rates evident (Tatem et al). Therefore, to transform theseecological niche, risk and transmission potential models into accurate estimates of burden that are neededto guide strategies and assess system demands, data on women of childbearing age, births and pregnanciesat similar levels of spatial disaggregation are required. The lack of such data has meant that existing studieshave been restricted to using simple national-level estimates to adjust gridded population datasets (Messinaet al, Perkins et al, www.worldpop.org.uk/zika/), ignoring spatial heterogeneities and likely leading toestimates that include substantial spatial biases.

Amount: £154,913
Funder: The Wellcome Trust
Recipient: University of Southampton

Epigenetic determinants of bone microarchitecture 02 Mar 2016

There is increasing evidence that associations between adverse early life exposures, for example intrauterine vitamin D deficiency, and postnatal skeletal development might be mediated by epigenetic mechanisms. Using a well-established discovery pipeline from genome wide to individual CpG sites and subsequent function, perinatal methylation at two genes (retinoid X receptor alpha gene (RXRA), essential in the action of 1,25(OH)2 vitamin D and other nuclear hormones; and CDKN2A, involved in cell senescence) has been shown to be associated with offspring bone mineral content at four years of age. However, it is unknown whether such associations persist into later childhood, whether they involve bone microarchitecture, and whether supplementation with vitamin D in pregnancy may influence these epigenetic marks. I aim to investigate whether DNA methylation at these two sites is associated with bone mass and microarchitecture at twelve years of age in the offspring of the Southampton Women’s Survey (SWS), and to test whether gestational supplementation with vitamin D, in a completed randomised controlled trial (MAVIDOS), leads to altered DNA methylation at these loci. Finally, I will use SWS data to undertake a genome-wide discovery approach to identify other CpG sites associated with bone mass and microarchitecture.

Amount: £252,397
Funder: The Wellcome Trust
Recipient: University of Southampton

Biomedical Vacation Scholarship. 22 Jun 2015

Not available

Amount: £11,750
Funder: The Wellcome Trust
Recipient: University of Southampton

Additional findings and WGS: patient, family members and health professional perspectives . 18 Sep 2015

Whole-genome sequencing (WGS) is being offered to NHS patients with rare diseases as part of the 100,000 genomes project - a hybrid clinical research venture. Clinical interpretation of sequence variants is improved by comparison with parental samples, so these will be analysed at the same time, wherever possible. Diagnosing the presenting disease is the primary aim, but parents will also be offered separate tests that predict other, possibly unexpected, conditions such as cancers/heart disease, or risks to future children. The results from some of these additional findings will only be disclosed if both parents are carriers of the same condition (couple results) since it is only then that future pregnancies would be at risk. These are all new practices for the NHS. The practical and ethical issues raised urgently need scrutinising. This project aims to sow the seeds for a collaborative award (CA-PIs Farsides/Williams/Parker/Lucassen) by capitalising on an early cohort of patients recruited to the 100,000 genomes project to identify research questions and methodology for the larger project. This work builds on our existing empirical ethics research on incidental findings and consent and confidentiality in genetic medicine, and thus forms a natural and timely progression.

Amount: £29,850
Funder: The Wellcome Trust
Recipient: University of Southampton

Health Law and Bioethics at the Frontiers of Innovation Postgraduate Bioethics Conference. 14 Jul 2014

The Postgraduate Bioethics Conference (PGBC) is an annual conference aimed at doctoral researchers in applied ethics broadly conceived. Over the past seven years, often with the support of the Wellcome Trust, PGBC has become established as the leading environment for doctoral candidates whose work involves bioethics to network and present their work. The theme of PGBC 2014 is Health Law and Bioethics at the Frontiers of Innovation and it will take place on 4-5 September at the University of So uthampton. There will be keynote speeches by Professor John Bryant, Professor Bobbie Farsides, Professor John Harris and Professor Jonathan Montgomery. With a focus on training and career development, we will also run two workshops. Bioethics in Practice will be led and organised by Professor Montgomery. Publishing in Bioethics , will involve a panel of editors from leading bioethics journals. Confirmed speakers are Professor Ruth Chadwick (Bioethics), Dr John Coggon (Health Care Analysis), P rofessor David Hughes (Sociology of Health and Illness) and Dr Sara Fovargue (Medical Law Review). We expect over 50 participants, of whom 24 will present a paper. The participants are likely to be mostly UK and Ireland-based doctoral researchers. Please see the website for further details: www.postgradbioethics.com.

Amount: £6,289
Funder: The Wellcome Trust
Recipient: University of Southampton

The Association of Medical Humanities Annual Conference 2014: The Art of Compassion. 23 Jun 2014

The Association for Medical Humanities (AMH) aims to progress the international culture of medical/ health humanities through research, scholarship and practice. Medical humanities include: (i) the use of the arts and humanities in medical/ healthcare practice and education, (ii) academic studies of the body in illness and health, and of medicine and healthcare, in historical, cultural and social contexts. Activities include an annual conference and the Association is linked to the BMJ Group jou rnal Medical Humanities. The annual conference provides an international interdisciplinary forum to exchange research, teaching, science and arts based enquiry into the medical humanities. This meeting will be the 11th conference of the AMH and hosted for the first time at Southampton. Previous conferences have been funded by the Wellcome Trust. Each conference takes a different theme and this year's theme is, The Art of Compassion, which aims to facilitate interdisciplinary exploration of me dical humanities within the context of medical education and patient care after the Francis report. Objectives are to share the extent to which medical humanities can generate understanding and maximise its potential to transform medical practice. Diverse speakers, papers and workshops will consider the nature of compassion from historical, philosophical, cultural and global perspectives

Amount: £5,000
Funder: The Wellcome Trust
Recipient: University of Southampton

Open access award 2013/14. 16 Sep 2013

Not available

Amount: £30,000
Funder: The Wellcome Trust
Recipient: University of Southampton

Biomedical Vacation Scholarship. 25 Jun 2012

Not available

Amount: £4,320
Funder: The Wellcome Trust
Recipient: University of Southampton

Biomedical Vacation Scholarship. 24 Jun 2013

Not available

Amount: £5,760
Funder: The Wellcome Trust
Recipient: University of Southampton

Southampton NMR Centre. 04 Feb 2010

The University of Southampton, as part of the development of the Institute for Life Sciences, is currently building a dedicated nuclear magnetic resonance (NMR) centre. Based on existing collaborations; the centre will bring together four groups from the Schools of Biological Sciences and Chemistry with expertise in NMR method and instrument development with those experienced in their application to biological systems. The centre s key objectives are: 1) To establish a state of the art experim ental NMR facility for the seamless integration of liquid and solid-state NMR for the acquisition of data on systems including soluble macromolecules, protein assemblies, biological membranes and fibrillar proteins. 2) To support a multidisciplinary research portfolio in the life sciences including the fields of amyloidogenic diseases, infectious disease, cancer science, membrane biology and drug development. 3) To facilitate the development of new NMR methods and technologies geared towards solving timely and fundamental problems in molecular cell biology 4) To promote an academic environment that fosters the exchange of ideas and sharing of expertise between different disciplines. 5) To provide a modern platform on which to train the next generation of researchers in the development and application of NMR to biological systems.

Amount: £77,846
Funder: The Wellcome Trust
Recipient: University of Southampton