- Total grants
- Total funders
- Total recipients
- Earliest award date
- 17 Oct 2005
- Latest award date
- 06 Dec 2005
- Total GBP grants
- Total GBP awarded
- Largest GBP award
- Smallest GBP award
- Total Non-GBP grants
We are investigating the mechanisms of alternative pre-mRNA splicing in three model systems in which the hnRNP protein polypyrimidine tract binding protein (PTB), plays a central role as a repressor. This programme will focus upon PTB, in particular analyzing its unusual role in effecting smooth muscle specific splicing of the a-tropomyosin gene, which contrasts with its more common role as a widely active repressor. · The first part of the proposal involves analysis of the mechanism of PTB-mediated repression of model pre-mRNAs. We aim to analyse the mechanistic basis of PTB-mediated splicing repression, identify additional regulators that are responsible for SM-specific regulation, analyze interactions between PTB and the cofactor raver1 that affect alternative splicing, and dissect the function of repressor domains in both proteins. · The second part of the proposal involves the application of quantitative proteomic approaches, as part of the current attempts to understand alternative splicing on a global basis. One approach will use proteomic approaches to characterize alternative splicing events that are affected by perturbations in the levels of PTB or other splicing regulators. The second application will be to characterize "cellular splicing codes" comprised of the relative levels of nucleoplasmic RNA binding proteins.
Computational human sensorimotor control. 20 Oct 2005
The main objective of this programme is to study the computations underlying planning and learning in human sensorimotor control. The programme will investigate three projects. Dynamic motor learning: The advent of robotic technology that can generate computer controlled force-fields that perturb movements has led to a dramatic increase of our understanding of sensorimotor learning. In this project we will develop novel robotic technology which will allow realistic simulations of virtual hand-held objects to examine dynamic learning in tool use. Probabilistic mechanism in sensorimotor learning: Sensory and motor uncertainty form fundamental constraints on human sensorimotor control. This project will further develop our work on Bayesian sensorimotor estimation and optimal control, focusing on how the CNS deals with uncertainty. Statistics of action: The statistics of sensory inputs, suchas natural scenes or sounds determine the way the CNS develops and represents the world. However, there is no data on the natural statistics of everyday movements and this project will be the first to examine the relationship between the natural statistics of actions and sensorimotor control processes. The overall goal is to integrate these three inter-related areas so as to provide a cohesive understanding of the computational processes involved in sensorimotor control.
Lentivirus genomic RNA encapsidation involves specific recognition of the low abundance genomic RNA in the infected cell by the viral Gag protein. It is dependent on RNA structures in the viral leader. Studies on these have elucidated several novel RNA motifs and RNA/protein capture mechanisms with relevance to both viral and cellular processes. Preliminary studies on FIV suggest that the packaging mechanism is very distinctive compared to that of HIV in that in FIV the process is dependent on two discrete regions in the leader separated by a redundant sequence. The aim is to map by mutagenesis and two and three dimensional structural analysis the extent and conformation of the packaging signal region of FIV. We predict that this will lead to novel data on RNA structures and on RNA/RNA and RNA/protein interactions of direct importance to normal cell function. It will also lead to an improved understanding of the packaging process in FIV. This will improve the efficiency and safety of the promising lentiviral vector system based on this virus.
Development of a simple, rapid and affordable HIV RNA test for early diagnosisof HIV-infection in infants and for antiviral therapy monitoring in resource-limited settings. 27 Oct 2005
Recent initiatives such as that of the World Health Organisation to provide 3 million HIV-infected individuals with antiretroviral therapy (ART) by 20051 will increase the availability of HIV treatment in developing countries. Such large-scale treatment projects will also require diagnostics and a means to monitor treatment to be fully effective. In the developed world, measurement of HIV load is standard for diagnosis and monitoring of HIV-infected individuals. Currently available HIV load assays are unsuitable for resource-limited settings because they are complex, time-consuming and require expensive instruments and test kits as well as cold-chain shipment and storage of reagents. The availability of a simple and robust test for the determination of HIV load will be crucial for the successful implementation of HIV treatment in the developing world.The objective of this project is to develop a simple and rapid HIV RNA assay designed specifically to assist health care workers in developing countries both with the early diagnosis of HIV infection, particularly in neonates, and for HIV therapy monitoring when appropriate quantitation standards are included. The proposed assays will be based on visual detection of isothermally amplified HIV RNA by dipstick. Rather than reporting absolute quantification of viral RNA, the assay will give readout of clinically relevant thresholds.
'What's in a name? - Authorship and Authority in the Transmission of Medicinal Recipes from 'Hippocrates' to 'Galen'. 10 Nov 2005
Names (real authorial names or pseudonyms) attached to ancient pharmacological treatises, or within a treatise, to individual recipes, conferred authority on pharmacological material. This project will investigate the relation between authorship and authority in the transmission of pharmacological knowledge in antiquity, taking into account variations through time, variations according to the literary genres in which recipes are listed, and variations according to the social origin of the authority named. I will study Greek and Latin sources from the second half of the fifth century BC (approximate date of the recipes preserved in the Hippocratic Corpus) to the end of the second /beginning of the third century AD (date of the pharmaceutical treatises in the Galenic Corpus). I will write five chapters covering the strategies used by pharmacological compilers to establish their authority; the methods of source quotations in pharmacological writings; the involvement, real or imagined, of political figures in pharmacology; the appropriation of traditional remedies by medical writers; and parallel versions of recipes attributed to individuals. Building upon my previous research on pharmacological recipes, and drawing upon recent studies on the history of the book, this study will enhance our understanding of the construction and transmission of ancient medical knowledge.
Long-term success to treat obesity is dependent on strategies aimed at preventing the concomitant fall in energy expenditure associated with food restriction (5). Energy modifications geared toward increasing energy expenditure may provide an alternative, independent means of promoting weight loss or, even more importantly, of preventing weight regain. In this project we investigate Bone morphogenic protein 8b (BMP8b) function as a novel mechanism regulating peripheral mechanisms of mammalian thermogenesis and fuel partitioning as strategies to prevent obesity. Our hypotheses are that: a) BMP8b may be an activator of thermogenesis and fatty acid oxidation and therefore a potential therapeutic target for obesity and type 2 diabetes mellitus. b) BMP8b may redirect the differentiation of white adipocyte fat pads towards more pro-thermogenic brown adipose fat pads. and c) BMP8b may promote energy dissipation in the skeletal muscle. The specific aims of this proposal are: 1. To establish the role of BMP8b on global energy homeostasis: thermogenesis, obesity and insulin resistance. 2. To establish the role of BMP8b in white and brown adipogenesis.3. Identification of signalling mechanisms mediated by BMP8b in adipocytes and skeletal muscle. These objectives will be accomplished through a multiple integrated approach involving in vivo and in vitro strategies
Regulation of smooth muscle cell (SMC) maturation by transforming growth factor (TGF)-b1 in development and disease. 06 Dec 2005
Smooth muscle cell (SMC) maturation is a critical determinant of vascular function. I recently demonstrated that mature SMC development from embryonic stem cells (ESC) is partly dependent on endogenous transforming growth factor (TGF)-?? signalling. However, it is unclear whether TGF-?1 acts directly on developing SMC and precisely how it regulates gene expression. My hypothesis is that direct TGF-?? signalling plays a key role in the differentiation/maturation of SMCs from ESCs, partly through epigenetic mechanisms involving histone modifications. First, I will test whether the requirement for TGF-?? signalling in SMC development from ESCs is cell-specific by generating murine ESC lines in which signalling will be up- or down-regulated in developing SMCs only. Then, I will identify histone modifications associated with the expression of SM-specific genes during development, determine whether these are modulated by TGF-?? signalling and whether altering histone modifications can regulate SMC gene expression. Finally, I will examine if ESC-derived SMCs, with constitutively increased TGF-? signalling, resist phenotypic modulation and inflammatory activation and thus assess their potential as atherosclerosis resistant substrates in future tissue engineering applications.
Inflammatory bowel disease genetics: candidate gene investigation by genetic association study and expression analysis in ulcerative colitis and Crohn's disease. 06 Dec 2005
Introduction: Inflammatory Bowel Disease (IBD) genetics is an area of rapid scientific progress. Crohn's disease (CD) and ulcerative colitis (UC) comprisethe main and related forms of IBD. Fine mapping of IBD linkage intervals has identified NOD2/CARD15 as a confirmed CD gene, and OCTN1, DLG5 and MDR1 have been recently implicated for CD, UC or both. Several additional strong linkages await resolution, with the prospect of fundamental advance in our understanding of IBD pathogenesis and the heterogeneity within CD and UC.Aim: Working with the world-class facilities and expertise in Cambridge, I aim to perform a systematic large-scale positional candidate gene association study to identify UC and CD susceptibility genes and phenotype-genotype correlations. I will then investigate expression patterns for genes implicatedin the association analysis in intestinal biopsies.Method: Using DNA and clinical data from 2000 UC subjects, I will genotype 768 non-synonymous or tagsingle nucleotide polymorphisms within two strong and replicated regions of linkage. I will then analyse genotypes to identify associated variants, as well as gene-gene and phenotype-genotype correlations. I will also perform replication analyses in an independent panel for CD-associated loci from the UK-wide 500 000 marker Welcome Trust Case Control Consortium study (WTCCC) forwhich the Cambridge group is playing a leading role.Expression patterns of genes identified for both UC and CD will be assessed using quantitative rtPCR,immunohistochemistry and RNA FISH in intestinal biopsies from cases and controls.
Schistosomes undergo a complex developmental lifecycle inhabiting two different hosts and alternating between free-living and parasitic forms. Studying when gene products are expressed will contribute to a greater biological understanding of this major pathogen useful in the identification of novel chemotherapeutic- and vaccine candidates. Although recent characterization of this helminth's transcriptome has yielded putative evidence related to the expression of approximately 13, 000 S. mansoni gene products, these estimates were based solely on comparative EST numerical profiling. As a more accurate measurement of relative RNA abundance, we recently profiled greater than 50% of this parasite's transcriptome using our well-characterized oligonucleotide DNA microarrays. While this important preliminary study provided semi-quantitative information related to the expression of greater than 7, 000 gene products, we still are lacking data related to the remaining 7, 000 or so transcribed sequences. In this grant proposal, we therefore want to extend and advance these significant preliminary findings by focusing on two specific aims: 1) develop an updated and near complete transcriptome long-oligonucleotide DNA microarray and 2) profile this DNA microarray with seventeen different schistosome life-stages/samples.
This year-long pilot study aims to resolve finally the developmental origins of gonadotropin-releasing hormone (GnRH) neurons in the hypothalamus and terminal nerve, which are critical for vertebrate fertility and reproduction. These neurons migrate into the brain along the olfactory nerves from the vicinity of the olfactory placodes, and a variety of fate-mapping and ablation studies have suggested they originate from the olfactory placodes themselves. However, a recent zebrafish fate-map has re-opened this debate by suggesting that cranial neural crest cells, which become intimately associated with the olfactory placodes, form at least some terminal nerve GnRH neurons, while the hypophyseal placode (which forms the anterior pituitary gland) forms hypothalamic GnRH neurons. The proposed series of quail-chick fate-mapping experiments, performed before neural crest cells reach the olfactory placodes, should settle the question of whether forebrain GnRH neurons in tetrapods are derived from neural crest cells, the hypophyseal placode or (as generally supposed) the olfactory placodes. This will resolve a long-standing debate and clarify our understanding of Kallmann's syndrome, in which GnRH neurons fail to reach the brain, resulting in hypogonadism. It will also provide an essential baseline for future experiments on the induction of GnRH neurons.
Trachoma, caused by ocular Chlamydia trachomatis infection, is the commonest infectious cause of blindness. We have shown that a single round of mass treatment with azithromycin can eliminate the infection from endemic populations for up to two years. The studies proposed here will enable this drug to be used to maximum effect. We will Determine the sensitivity and specificity of a new rapid point-of-care (POC) diagnostic test for ocular C. trachomatis infection in communities with high, medium and low prevalence of trachoma in three countries, before and after mass treatment. Use the above data, datasets from our current Wellcome Trust-Burroughs Wellcome Fund grand, to develop optimal sampling strategies to assess the need for community-based mass treatment depending on the prevalence of clinical signs of trachoma or of ocular C. trachomatis infection, determined by the POC test. Use this data to model the impact and cost-effectiveness of strategies in which a POC test is used to decide whether a community should be treated, and to compare this with the cost-effectiveness of the current strategy, based on the prevalence of clinical signs of trachoma.
The evaluation of genetic and phenotypic diversity in field isolates of Salmonella enterica serovar Enteritidis in Uruguay. 25 Oct 2005
Salmonella Enteritidis causes food-borne disease world-wide. It commonly contaminates the food chain and poultry products are a major source. Particular strains are associated with outbreaks. We have shown that in Uruguay different strains survive in the chicken population, with only a small proportion of them spreading through food to humans. We therefore hypothesize that different strains have different genetic characteristics that give rise to populations able to exploit different ecological niches. We thus propose to undertake comprehensive comparative genomic analyses of Uruguayan field isolates. Molecular typing methods will be used to allow sub-grouping of the strains. Multi-Locus Sequence Typing and microarray analysis will be applied to identify evolutionary trends, population structures, and genetic heterogeneity between strains. The genomes of three individual strains will be sequenced using new high-throughput technology at the Sanger Institute. This population information will inform selection of representative strains for use in in vitro and in vivo assays to assess their capacity to interact with different hosts.This study will provide unique information regarding the relationship between strain variation and the exploitation of different hosts by S. Enteritidis, and may inform the development of intervention measures that may be effective in developing countries.
Neutrophils become 'primed' after exposure to inflammatory mediators and this greatly enhances their subsequent secretory activity. Priming also alters neutrophil shape/deformability, integrin expression and longevity and hence has a profound affect on their rheological, adhesive and survival properties. Priming is a prerequisite for neutrophil-mediated tissue injury and plays a fundamental role in the pathogenesis of ARDS. Since priming is reversible, I will test whether the pulmonary capillary bed and/or spleen can trap and de-prime systemically-primed neutrophils before releasing them back into the circulation, and whether failure of this function results in neutrophil-mediated lung injury.Specifically, I will address:(i) What is the bio-distribution, preferential disposal site and circulating half-life of primed versus un-primed neutrophils? This will be addressed using 99mTc and 111In-labelled primed/un-primed neutrophils in healthy volunteers.(ii) Can pulmonary capillaries or the spleen retain circulating primed neutrophils, de-prime them and return them to the circulation? I will determine the capacity for neutrophils primed in vivo to de-prime ex-vivo and use flow-based techniques to track the intravascular fate of permanently or transiently primed 111In-labelled cells.(iii) What is the effect of lung vascular (ARDS) and airspace (pneumonia) inflammation on transpulmonary primed neutrophil gradients and does neutrophil transit time and pulmonary retention fraction predict the evolution of ARDS.