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Mycobacterial infections are a global concern in humans and animals. In the UK, tuberculosis (TB) in animals is not confined to cattle and badgers; it is a serious concern in deer, camelids and cats. We have shown that cats are significantly at risk of TB as ~1% of feline biopsies have changes consistent with mycobacteriosis; 15% due to M. bovis, 19% M. microti (the vole/rodent bacillus). Some M. bovis infections progress extremely rapidly - a major concern given the newly recognised cat to human zoonotic risk, and notifiable status (M. bovis spreads between many different mammalian species). We hypothesise that the outcome of a cats exposure to Mycobacteria is determined by the nature of the pathogen and the host's immune response. However, a major bottleneck is failure to culture in ~50% of cases (despite having ZN+ bacteria). Rapid diagnosis is key to determining zoonotic risk and management options. Currently, identifying the Mycobacteria species depends on a) detecting ZN+ bacteria b) IFN-gamma ELISA c) culture. These tests can detect Mycobacteria and the ELISA differentiates (with some specifity) M. bovis, M. microti and M. avium. Culture is a gold standard method to confirm live bacteria but is lengthy and frequently negative (despite tissue being ZN+). To rapidly identify infecting bacteria will impact significantly on knowing the zoonotic, inter- and intra-species risk, management options, affect survival (likelihood of clearance or latency) and inform ongoing monitoring. It could also monitor in-contact animals. We need to invesitgate the host respone to these infections if we are to understand Mycobacterial diseases more fully, and so elucidate the role of the host's response in determining which cases are likely to spread disease or, where appropriate, respond to treatment. The project has 5 objectives: 1) Collect samples from Mycobacteria-infected cats and controls. Confirm Mycobacterial presence using current tests (culture, ZN, IFN-gamma ELISA) (RDSVS, Roslin, Biobest). 2) Isolate Mycobacterial DNA using established protocols from fresh frozen tissue or FFPE blocks. Determine the Mycobacterial sequences using the latest second and third generation sequencing methods (Illumina HiSeq 2500/MiSeq; Oxford Nanopore MinION) and compare them to published genome sequences (Roslin/Edinburgh Genomics). 3) Perform comparative analysis of the genome sequences of the isolated strains to identify sequence similarities and differences. Identify unique sequences for the development of strain-specific PCRs and develop multiplex PCR assays for the rapid diagnosis of feline TB and other Mycobacterial infections (Biobest). 4) Monitor cases long-term to correlate specific sequences with disease outcomes, and any transmission (RDSVS). 5) The nature of host-pathogen interaction between macrophages and Mycobacteria defines infection outcome in a number of species, but has not been investigated in cats. Understanding the basis for immunity or disease progression will inform veterinary medicine, and help to suggest appropriate management regimens for infected cats. We will identify cats with M. bovis TB and compare macrophage distribution, function and phenotype between infected and control tissues (Roslin). This is a new collaboration bringing together expertise in feline medicine and tuberculosis, with macrophage biology, and pathogen genome sequencing incorporating both Roslin/RDSVS and Biobest Laboratories.

Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.

Doctoral Training Partnerships: a range of postgraduate training is funded by the Research Councils. For information on current funding routes, see the common terminology at https://www.ukri.org/apply-for-funding/how-we-fund-studentships/. Training grants may be to one organisation or to a consortia of research organisations. This portal will show the lead organisation only.

In humans numerous cancer-associated mutations in Notch have been identified in different tumours and genetic diseases such as the dementia causing CADASIL disease. The links between the mutations, the affect on Notch signalling and misregulation arising in CADASIL are poorly understood because we lack sufficient understanding of functional contributions of large portions of the Notch receptor to its regulation. Through Drosophila genetics many mutations of Notch have been identified which cause an up or down regulation of Notch signalling, which have been linked to developmental phenotypes in the fly and whose mechanism of misregulation is also unclear. Some of those mutations such as the nd3 allele, resemble the kind of mutation identified in CADASIL patients. Combining these human and Drosophila data sets there is considerable resource of information regarding the locations of different functionally important regions of the Notch receptor structure. This project will combine studies in human and insect cell culture, and in vivo in Drosophila tissues, to understand the fundamental mechanisms of activation or inhibition resulting from different Notch mutations to provide a basic understanding of Notch structure and function and how it relates to the regulation of Notch trafficking which plays a key role in controlling the level of signalling.

Filamentous plant pathogens are a group of eukaryotic pathogens including oomycete genus Phytophthora, as well as rust fungi, and Powdery mildew fungi, which cause most destructive plant diseases and threaten global food security. These pathogens cost billions of dollars annual looses to modern agriculture and severely impact subsistence agriculture in developing countries. These microorganisms are accommodated within the host cells through specialized cellular structures. However, little is known about molecular mechanisms underlying microbial accommodation inside the plant cells. The proposed research aims to characterize the host processes required for accommodation of filamentous plant pathogens inside the plant cells with a specific focus on illustrating the role of plant endomembrane transport system in this process. Among the oomycetes, Phytophthora spp. cause some of the most destructive plant diseases in the world and cause enormous economic damage to important crop species such as potato, tomato, and soybean, as well as environmental damage in natural ecosystems. Phytophthora infestans, the Irish potato famine pathogen, which causes late blight of potato and tomato, is one of the most important biotic threats to global food production leading to worldwide economic losses exceeding $5 billion annually. Unfortunately, late blight of potato is a reemerging destructive disease, which caused severe epidemics in potato farms across the UK lately. Our long-term objective is to dissect the molecular mechanisms underlying plant cell autonomous immunity and the role of endomembrane traffic in this process by particularly using P. infestans as a model pathogen. Like some other filamentous pathogens that form destructive plant diseases, P. infestans is accommodated inside the host cells by forming specialized compartments termed haustoria, which are separated from the invaded plant cells by newly synthesized host derived membranes with unknown origin and composition. This interface is critical for development of parasitic infection by enabling efficient macromolecule exchange. Therefore, understanding the regulation of macromolecule exchange processes at host pathogen interface is critical to develop novel strategies to engineer disease resistance in plants. Unfortunately, despite being discovered decades ago, our current knowledge about haustoria is limited. Deciphering the cellular and biochemical activities at this interface is critical for understanding the mechanisms of pathogenesis. Like other pathogens, P. infestans secretes a battery of proteins, termed effectors that suppress plant immunity and enable parasitic infection. Recently we discovered that some of these effectors specifically accumulate at the host pathogen interface inside the infected plant cells. In this study we aimed to identify host components of focal immunity using one such effector, termed PexRD54, as a molecular probe. Our preliminary work unraveled that around haustoria, PexRD54 associated with a host protein named Rab8-1, a member of Rab GTPases family of eukaryotic vesicle trafficking regulators. Our goal is to functionally characterize PexRD54 and Rab8-1 in order to establish the roles of these molecules at host pathogen interface. This study will help to establish functional characterization of plant endomembrane transport processes perturbed by pathogens and help understanding the origin, function and composition of the host pathogen interface. In turn, a detailed knowledge of PexRD54 manipulation of Rab8-1 will improve our understanding of the microbial accommodation inside the plant cells and will allow for creating renewed opportunities for engineering disease resistance in crops.

Styrene is an important chemical across multiple industries, most notably as a monomer in production of plastics and rubbers. There has been demonstrated efforts to produce styrene using an E. coli platform, however the toxicity of this molecule significantly reduces productivity. My research aims to apply experimental evolutionary techniques to an E. colistrain to create mutant variants that display desirable styrene tolerance behaviour. These variants will then be subjected to the 'omics' studies to determine the genetic, proteomic and the lipodomic basis of adaptation to exogenous styrene stress.

Parasitic plants are plants that rely entirely or partly on another plant in order to survive and reproduce [1]. In Africa, two parasitic plants causing particularly high levels of damage to cereal crops are Striga hermonthica and Striga asiatica. Both of which attach to the roots of their host plant and cause massive reductions in yield [2]. The development of resistant cultivars is seen as an important part of the strategy to tackle these parasites, but progress has been hampered, in part, by a lack of understanding of the molecular genetic basis of the interaction between the host and parasite [3]. To this end, it is important to better understand the molecular basis of resistance in the host and of virulence in the parasite. The latter is the broad aim of this project. I aim to use a combination of genomics, transcriptomics and bioinformatics to compare and analyze different isolates of the root parasitic plant Striga asiatica at the molecular level, with the goal of selecting candidate genes that may function as effectors for parasite virulence. Effectors are proteins secreted by parasites, which are predicted to modify the function and/or structure of the host in order to facilitate parasite fitness [4]. The discovery of such proteins from parasitic pants would open new avenues of research that would likely lead to the identification of novel resistance genes in host plants that could be invaluable to crop breeders. Initially, the genomes of virulent and avirulent isolates of S. asiatica will be compared for the presence/absence of genes, when growing on particular rice cultivars. This initial analysis will be followed up with a bioinformatics pipeline that will annotate the genomes according to known effector-properties from other pathosystems, in order to generate a list of potential candidate effectors. This list will be refined by integrating transcriptomics data, which will provide information on genes differentially regulated in the infection organ of the parasite (called the haustorium). A selection of the most promising candidates will be functionally analyzed in order to determine; (i) whether the gene plays a role in virulence and (ii) if a role in virulence is detected then further analysis will aim to characterize precisely how it functions.

The twin objectives of RegulaTomE are to determine the importance of transcriptional regulation of the metabolic pathways defining quality traits in tomato and to identify such transcriptional regulators at the molecular level. The selected quality traits include antioxidant capacity which impacts shelf life and nutritional value as well as traits determining fruit flavor and over-ripening which influence organoleptic properties and shelf-life. RegulaTomE will use the natural variation available in introgression lines (ILs) resulting from wild species crosses to tomato to assess the importance of transcriptional regulation, identify additional regulatory genes and assess underlying genetic and epigenetic variation. RegulaTomE will assess the potential for direct or indirect use of natural variation from an untapped wild species resource for crop improvement. To identify genes regulating metabolic pathways using the Solanum lycopersicoides ILs, and to capture genetic and epigenetic variation for application to gene discovery and tomato improvement, resources need to be developed, including a genome reference sequence for S. lycopersicoides and metabolite, DNA methylation and transcriptome profiles of IL fruit. RegulaTomE will lead to regulatory gene identification and new tools for metabolic engineering of fruit quality. The natural variation in fruit quality revealed by the S.lycopersicoides ILs could be applied to tomato improvement either directly through introgression into cultivated varieties or indirectly through the identification of target loci and corresponding allelic variation making positive contributions to quality traits within S. lycopersicum breeding germplasm. The outputs of RegulaTomE will provide a framework of understanding as well as tools, in the form of genes, target loci and molecular markers, to support development of longer shelf-life, more nutritious and more flavorsome fleshy fruits in other horticultural crops.

Doctoral Training Partnerships: a range of postgraduate training is funded by the Research Councils. For information on current funding routes, see the common terminology at https://www.ukri.org/apply-for-funding/how-we-fund-studentships/. Training grants may be to one organisation or to a consortia of research organisations. This portal will show the lead organisation only.

The purpose of this research is to understand how and where in the body the novel hormone fibroblast growth factor 21 (FGF21) acts to reduce food intake, decrease body weight and regulate body fat. This hormone was first discovered over ten years ago, but we are uncertain about which tissues in the body produce it, where it acts, and what its normal role in our biology is. Our previous research has exploited seasonal cycles in body weight in the Siberian hamster, as this provides a natural animal model of body weight gain in summer (fat state) and loss in winter (lean state). Using this model, we have already found that FGF21 is more effective at reducing appetite and causing weight loss in seasonally fat hamsters. This is a hugely important finding, because responses to other major metabolic hormones are often decreased in states of high body fat. The fact that obesity is an insulin- and leptin-resistant state presents challenges for using these pathways to manage body weight disorders. Understanding the natural biology of FGF21 should therefore have important implications for pharmaceutical and/or nutritional treatment of obesity as this pathway is likely to be amenable to manipulation. The first objective is to determine which tissues respond to FGF21 treatment by changing their uptake of glucose and fatty acids. This will be achieved using a small animal positron emission tomography (PET) scanner, which allows uptake of these metabolites to be observed non-invasively in living animals. We will also test in vitro whether FGF21 can promote fat breakdown. These studies will identify which tissues are the primary targets of FGF21 action, and confirm whether actions on glucose and fatty acid uptake underlie the whole body effects on fat depots and body weight. The second objective is to investigate the hypothesis that FGF21 also acts in the brain to reduce food intake and to increase energy expenditure. Other metabolic hormones such as leptin and ghrelin are known to signal from fat and the stomach respectively to the brain to regulate our appetite. Our preliminary studies also provide evidence that FGF21 can act in the brain. We are particularly interested in actions on a layer of glial cells in the hypothalamus known as tanycytes, as these cells express a receptor for FGF21 known as FGFR1c, and show changes in gene expression and glucose-stimulated calcium signalling in response to stimulation of these receptors. We will test the hypothesis by determining whether administration directly into the brain of FGF21 itself or a closely related compound developed by the pharmaceutical company Eli Lilly changes appetite, energy expenditure and body weight. We will also use imaging of slices of brain derived from rodents to determine whether FGF21 directly affects neurons and glial cells. The overall outcome of this project is that we will understand how the hormone FGF21 is able to produce its beneficial effects of improved glucose dispersal and loss of body fat. We will have identified which tissues respond to FGF21, and will determine if part of its action is in the brain via the control of behaviour and the autonomic nervous system. There are many beneficiaries of this project. The information gained will be important for other academic researchers in universities and in research institutes, and for researchers in the pharmaceutical industry working on obesity. In addition, the project will provide training in advanced imaging and experimental physiology, and the researchers will promote public understanding of research into appetite control and obesity.