Our organisms are powered by biochemical reactions from molecular machines often made up of individual proteins or protein complexes. Proteins are responsible for tens of thousands if not millions of molecular interactions either with each other (protein-protein interactions) or with small molecule ligands, metabolites and drugs (protein-ligand interactions). Measuring these interactions precisely and in high-throughput is a major challenge and forms the basis for our understanding of how biological molecular machines work and for imagining new ways to develop therapeutic drugs to treat diseases. We would acquire a Dianthus instrument that can precisely and robustly measure protein-protein and protein-ligand interactions in solution where optimal conditions for protein stability can be maintained. The instrument can simultaneously use two detection modes based on two different biophysical techniques: a spectral shift technology and a temperature related intensity change (TRIC) technology. Each has its own advantages, and the combination is really powerful in improving detection sensitivity (allowing us to use less of the precious materials we make) and can signpost potential "bad" reactions that need repeating or are untrustworthy. Doing experiments in a high-throughput format with a 384-well plate measuring >1000 reactions per hour will significantly accelerate our ability to discover new molecules as tools to understand biology and disease conditions. It will transform the way we discover and optimise drug candidates for early-stage drug discovery programs. We use multiple screening platforms to identify drug candidates and have a combination of techniques and expertise to optimise "hit" compounds. However, we lack a technology that can provide affinity measurements and rank our molecules according to their ability to bind or "stick" to our biological targets. The Dianthus instrument will bridge this crucial, informative gap in our discovery pipeline. Many of our projects will benefit from this technology. New enzyme families and drug candidates in the ubiquitin proteasome system promise to benefit inflammatory diseases (e.g. Lupus, Sclerodema, Rheumatoid Arthritis) and multiple types of cancer. These are hard targets because as large multimeric complexes they are difficult to produce and assay with current technology. Similarly, self-assembling systems that form protein fibrils and contribute to amyloidosis in Alzheimer's and type II diabetes are hard to study with current techniques. The new instrument will make it easier to discover molecules that prevent fibril formation. We have teams working on challenging targets that control protein folding and energy flow in mitochondria with promising molecules that may benefit patients with an aggressive and hard-to-treat brain cancer (glioblastoma). We made huge progress with membrane proteins that are targets for malaria, antibiotic resistance and heart disease and are developing new ways to drug these proteins. The 384-well plate system is also ideal for a ligand-discovery platform developed in Leeds which also performs chemical reactions in 384-well plates, ensuring perfect compatibility. Finally, we are using Dianthus to understand fundamental biology by measuring interactions of protein complexes with DNA to study how DNA damage is repaired. This will further our knowledge of disease biology and help us unveil a cell's "Achilles hill" to unlock new therapeutic targets in cancer. At time of writing, no UK institution has installed this technology. As such, the Dianthus instrument will be an amazing unique addition to our multiuser research facilities and complement our current capabilities. The instrument will also be hugely beneficial to the Leeds science ecosystem and local universities through our equipment sharing portal for northern universities and other interested institutions.

There are two major scientific challenges to address if the PEP-GAG gels are to be successfully translated to clinic. Challenge 1: To develop methods to effectively visualise the location of the gel, and identify if/where it disperses to over a period of time. In order to assess the long term efficacy of the treatment, it is necessary to be able to quantify the degree to which the gel disperses out of the nucleus over time, in both in vitro laboratory tests and in vivo. We have successfully used clinical radio-opaque agents to visualise the immediate location of the injected gel under x-ray/CT, but longer term, the agent and gel will not necessarily diffuse at the same rate. There have been some initial attempts to chemically bond markers to the gel, but further investigation is needed to examine if this affects their performance. Recent work has also examined the use of histological methods to identify the peptides which have shown some promise. In parallel, we are continuing to develop in vitro testing methods to examine the performance of the PEP-GAG gel under cyclic loading in the laboratory and have a planned in vivo study due to commence next year. If successful methods for visualising the gel can be developed, then greater evidence can be generated on the efficacy of the treatment and its likely longevity. If the gel can be seen long-term in vivo, then this will also reassure clinicians on how patients could be monitored. These aspects are critical if the PEP-GAG gel is to be successfully commercialised and adopted. Challenge 2: To identify the most suitable patient characteristics for the treatment to be successful and the optimum volume of PEP-GAG gel to inject. Different levels of disc degeneration cause changes to both the annulus and nucleus of the disc, and it is not yet known what stage of degeneration the treatment would be most effective, or what the important contra-indications would be. Our early laboratory findings have shown variance from specimen-to-specimen, likely due to the differences in the volumes of PEP-GAG injected compared to the degree of degeneration. Due to the large number of unknowns, this challenge is best addressed using computational models in which different variables can be systematically altered to evaluate their effect. We have developed finite element (FE) models of the disc to assess the mechanical performance, but the models do not yet have the sufficient sophistication in terms of representing the biphasic behaviour and gel-disc interactions to fully answer these questions. Addressing this challenge will provide underpinning evidence necessary for the PEP-GAG gel to progress successfully through clinical trials, where it will be essential to be able to identify exactly which patients will benefit and carefully regulate the procedure to avoid the risk of complication. These challenges require a multidisciplinary approach encompassing aspects of mechanical engineering and simulation alongside chemistry, biology, imaging and image processing The aim of this project is to optimise aspects of the PEP-GAG hydrogel nucleus augmentation procedure so that it can be more effectively translated to clinical practice.

The food system feeds societies, employs billions of people, and underpins international aspirations such as the United Nations Sustainable Development Goals. However, it is responsible for one-third of anthropogenic greenhouse gas emissions, threatens more species than any other activity, uses over half of all nitrogen and phosphorus, and over 70% of all freshwater. Balancing these benefits and pressures is essential for human wellbeing and the liveability of both specific places and the planet. These environmental pressures come disproportionately from the production of animal products (e.g. 57% of food system emissions come from livestock production, Xu et al 2021), largely due to their low environmental efficiency compared with plant-based foods (Poore & Nemecek 2018). Reducing consumption of animal products in wealthy countries and avoiding shifts to high-meat diets as economies develop is therefore essential to maintain a liveable environment. However, attempts to reduce consumption may be resisted due to the social and cultural importance of animal products and production systems, strong relationships between wealth and consumption, suggesting aspirational consumption (Tilman et al 2011), and political aversion to any perceived limiting of consumer choice. In this context, alternative proteins (APs) offer a potential solution. APs include plant-based, cultivated and fermentation-made meat, eggs, dairy, and seafood, which have significantly lower environmental impacts than animal-based counterparts. They may also offer health benefits at the individual level (e.g. higher fibre, lower calorie density) and societal level (e.g. reducing risks from zoonotic diseases and antimicrobial resistance). Importantly, they may enable consumers to substitute lower-impact products into diets while continuing to access familiar tastes and dishes. However, the growth of APs has not been universally welcomed, being described as a technological solution that fails to address the complex social, economic and cultural factors associated with food production and consumption. In addition, the social, cultural, economic, and ecological transformations that could result from a widescale shift to APs are understudied. Instead, most research has focused on technical product development or life cycle assessments of specific products. Given the speed and scale of AP development there is an urgent need to address this knowledge gap. This interdisciplinary project has been co-designed with the European team of the Good Food Institute-the world's leading AP-focused international third-sector organisation-to answer the overarching question: "How will alternative proteins affect people and the environment in the UK and Europe over coming decades?". To do so, the student will tackle three linked questions: (1) How are different APs likely to be accepted in different European contexts? (2) How could European demand for different proteins change up to 2050? (3) What would the social, economic, and environmental impacts of meeting this demand be? This interdisciplinary project will make empirical and theoretical contributions to a range of disciplines, including economics, land economy, and environmental social sciences. For example, providing insight into the substitutability of different protein sources across cultures; the liveability implications of different ways of meeting future food demand; and how shifts to APs could affect people's relationships with their environments.

The project aims to study the features of the Hepetitis E Virus (HEV) genome which permits replication between species. HEV has eight genotypes however we are interested in the differences between genotype 1 and genotype 3. Genotype 1 is an obligate human pathogen meaning it is only capable of human transmission, primarily via the fecal-oral route. Genotype 3 is zoonotic and commonly infects deer, pigs and wild boar. It is typically transmitted to humans through infected meat. Using viral and replicon systems we will utilise saturating mutagenesis within the hypervariable region (HVR) of the HEV genome. We can then insert the replicon RNA into human and porcine cells and compare the replication abilities using fluorescence tagging and live cell imaging. These efforts will initially focus on Genotype 3 as the literature suggests Genotype 1 is not replication competent in porcine cell lines. However, once controls are established, there is scope to mutate and compare the HVR of Genotype 1 to establish which areas permit cross species replication of HEV and related viruses.

Currently there is little understanding of how microorganisms in water systems are dispersed into the environment and the potential risks associated with this. The proposed project aims to model the release of microorganisms from showers to quantify occupant exposure. This will involve particle and bioaerosol measurements from an experimental shower system at PHE and development of a CFD model to simulate the dispersion of water droplets and microorganisms from the shower head. Combing the data of these two elements will enable the modelling of the evaporation and transport of droplets in a bathroom environment. In this way the influence of parameters such as temperature, humidity and ventilation on the fate of microorganisms within the droplets.