In this thesis gallic acid-triethylene glycol (GATG) dendrimers were synthesised and efficiently functionalized with hydroxyl groups, phenylboronic acids and primary amines. The interactions of the dendrimers with bacteria and the potential for development of new antimicrobials were evaluated in this study. Specifically, the ability of the dendrimers to induce bacterial clustering and interfere with small molecule autoinducer-2 (AI-2) in the Quorum Sensing (QS) pathway of the marine bacteria V. harveyi was studied with the use of Coulter Counter aggregation assays and detection of QS–controlled luminescence. Novel alkynylated ligands with diol-, tetraol-, glucose- and mannose- moieties were synthesised and successfully functionalized to GATG dendrimers of generation G1 and G3 through catalyst-free azide-alkyne cycloaddition (AAC). The results of luminescence experiments reveled that the dendrimers functionalized with hydroxyl groups decreased AI-2 induced luminescence of V. harveyi MM32 at the at early time points (4 h) while a dose-dependent increase of luminescence and increased bacterial growth was observed at later time points. GATG dendrimers of generation G1 and G3 were decorated with 9 and 81 phenylboronic acid in the periphery. These dendrimers had an inhibitory effect on growth and luminescence as observed by luminescence, aggregation and colony forming unit-counting assays. Although the mechanism is not yet fully understood, these promising results should be further explored. Cationic GATG dendrimers of generation G1, G2 and G3 with 9, 27 and 81 primary amines in the periphery induced formation of clusters in V. harveyi in a generation dependent manner, an improved ability to induce cluster formation when compared with poly(N-[2- (dimethylamino)propyl]methacrylamide), a cationic linear polymer previously shown to cluster bacteria. Viability of the bacteria within the formed clusters and the evaluation of the QS controlled luminescence suggests that the GATG dendrimers may be activating microbial responses by maintaining a high concentration of QS signals inside the clusters while increasing permeability of the microbial outer membrane. Thus, a generation-dependent effect in bacterial luminescence production and membrane permeability was induced by the cationic dendrimers. The inhibition of growth and increased membrane permeability in combination with cell clustering may be promising antibacterial features of these dendrimers. These results highlight the potential of the GATG dendritic platform to develop new antimicrobials aimed to target microbial viability and/or virulence (e.g. adhesion) and encourage further investigations on the importance of polymeric architecture and multivalency in the antimicrobial field.

Acute pancreatitis (AP) is a leading cause for hospitalisation and has significant quality of life implications for the patient and cost implications for the National Health Service. Although most episodes of AP are mild and self-limiting, the severe form of the disease is associated with a high mortality. In the absence of definitive treatment, management is mainly supportive. There is an urgent need to develop more effective biomarkers and drugs to manage AP. Genome-wide studies have demonstrated that proteins that bind to heparin (HBPs) form highly interconnected networks which are functionally important in health and disease. It was hypothesized that this is true in the pancreas and in AP. Testing this hypothesis, using mRNA as a proxy for protein, it was shown that HBPs constitute an important extracellular sub-proteome within the normal pancreas and in major pancreatic diseases that is likely to provide a rich repository of potential biomarkers and drug targets. Building upon this work, a proteomic analysis of HBPs in normal pancreas (NP) and in caerulein-induced mouse AP was undertaken. This has more than doubled the number of HBPs to 883, with 460 new HBPs identified. These may represent the most interconnected set of extracellular proteins and therefore with the greatest regulatory potential. Non canonical HBPs such as NDUFS4, NDUFS6, NDUFS7, NDUFS8, NDUFA9, NDUFA10, NDUFA9 and NDUFA10 were identified and found to be underexpressed in AP as compared to NP. These may have potential moonlighting roles, not previously known. By virtue of being extracellular and binding to heparin, HBPs are accessible and are potential biomarkers and drug targets in AP. In addition to identifying existing biomarkers in AP such as pancreatic amylase, a number of HBPs with biomarkers potential such as HRG, CD14 and FN1 were identified and need further investigation. HBPs such as SERPINC1, VEGFA and PIP5K1C need further evaluation in drug development. These along with modified heparins, heparin mimetics and matrix therapy in AP provide exciting areas for future research.

HIV continues to be a global healthcare challenge, with estimates suggesting there are currently 35.5 million infected people globally. To date, there have been an estimated 36 million HIV/AIDS related deaths worldwide. The story is changing however, with the advent of Highly Active Antiretroviral Therapies (HAART), allowing patients to live to normal life expectancies and with increasingly better quality of life. This, coupled with the fact that there has been a 40-fold increase in the number of people with access to antiretroviral therapy, has led to a 29% reduction in AIDS related deaths since 2005. Despite this encouraging data, there are still numerous limitations of antiviral therapy, including poor bioavailability, poor patient adherence, and emerging resistance. It is hoped that nanomedicine may offer a route to alleviating some of these issues by achieving an equal therapeutic concentration of drug but with a lower dose. The aims of this thesis were to develop a novel nanoemulsion based formulation of EFV and LPV and to assess the suitability of this formulation as dosage form. Nanoemulsions can be stabilised by surfactants, but often this can have unwanted safety profiles. Stabilisation can also be achieved using amphiphillic polymers that can be synthesised using biocompatible monomers like ethylene glycol. Chapter 2 demonstrates the synthesis of Ethylene Glycol based polymers using both conventional free radical and Atom Transfer Radical Polymerisation techniques. Nanoemulsions have previously been shown in the literature to increase the bioavailability of poorly water-soluble drugs, Chapter 3 shows the development and optimisation of an Oil-in-Water nanoemulsions. The data showed that nanoemulsions synthesised with volatile cosolvents were able to achieve sub 300 nm diameters and have good long-term stability. Increasing the accumulation and permeation of a poorly water-soluble compound should lead to improvements in bioavailability. Chapter 4 shows that the optimal nanoemulsion had comparable accumulation to aqueous solutions and superior apparent permeability cross Caco-2 cell monolayers. The antiviral activity was equipotent to the aqueous solution, as shown in Chapter 5. This confirms that nanoemulsion did not prevent the API from reaching its sight of action. Finally, all new formulations have the potential for detrimental side effects and immunological responses. It is therefore necessary to conduct pre clinical studies to predict such occurrences. Chapter 6 details the lack of immunological response seen in nanoemulsions, but highlights potential interactions with coagulation. In conclusion, this study has found that polymer stabilised oil-in-water nanoemulsions based on Castor oil have promising safety and pharmacological profiles. Further in vivo and studies are now warranted in order to further predict the suitability of nanoemulsions in man.

Advancing clinical reasoning skills is one of the main outcomes of postgraduate master's level (M-level) programmes approved by the Musculoskeletal Association of Chartered Physiotherapists (MACP). While, the outcomes of these programmes were investigated in multiple retrospective studies, there is a limited understanding of the learning culture that drives change. Thus, the aim was to examine the learning culture of an MACP approved programme to capture the sociocultural mediators that advanced clinical reasoning skills. An empirical longitudinal mixed-methods theory-seeking case study was conducted over a period of 18 months. Participants included seven educators and six students. Data analysis was premised on the methods of a Constructivist Grounded Theory. Gradual and progressive advancement of clinical reasoning skills was identified. A model of a culture of convergence and synergy was constructed to conceptualise the relationship between students, the programme and the wider context. It demonstrates the value of convergence and synergy in supporting professional learning. This novel conceptual understanding of advancing clinical reasoning through M-level education suggest that pedagogues need to actively seek to create a culture convergence and synergy to achieving successful learning outcomes. The context-bounded knowledge provided in the thesis aid pedagogues to better design M-level curriculums.

The \(Escherichia\) \(coli\) enzyme nitroreductase has been proposed as a candidate for the Gene-Directed Enzyme Prodrug Therapy approach in treating cancer. Structural studies on the enzyme were instigated in a first step towards improving enzyme activity. The enzyme was crystallized with the substrate analogue, nicotinic acid, and the structures of three crystal forms obtained. The fold has a mixed a/P structure, with a molecule of nicotinic acid bound next to the FMN cofactor. Several active site residues were identified as candidates for mutation. This procedure produced many mutant enzymes with increased catalytic activity. One double and four single mutants were chosen from these and crystal structures determined. The resulting information from this, and the establishment of a proof of principle, provides the basis for iterative cycles of enzyme improvement. The human hydroxysteroid dehydrogenase AKR1C3 has been proposed to play a role in prostaglandin metabolism. Its inhibition by non-steroidal anti-inflammatory drugs may be important in a tumour differentiation strategy. AKR1C3 was crystallized, and the structure solved with bound nucleotide cofactor and several inhibitors, including the drugs indomethacin and flufenamic acid. Having obtained information on drug binding to AKR1C3, selective inhibitors can be designed, avoiding inhibition of "housekeeping" enzymes such as cyclo-oxygenases.

This thesis explores Bayesian methods for the statistical design, analysis and synthesis of clinical trials, and compares these with a frequentist approach in several settings to determine key differences, advantages and limitations. A review of randomised trials indicates that Bayesian methods are rarely applied, but useful for making probability statements and incorporating prior evidence, especially in trials with small sample sizes. These advantages are illustrated in a trial in congenital lower urinary tract obstruction, which has few events but elicited prior distributions about treatment effect. Bayesian methods are then developed for meta-analysis of phase II trials and multiple outcomes, with an emphasis on informing phase Ill trial decisions. A Bayesian random-effects logistic regression is advocated for meta-analysis of a binary outcome to account for all parameter uncertainty, and to derive prediction intervals for the treatment effect in a new phase III trial. Bayesian multivariate meta-analysis methods arc then encouraged to make joint inferences across multiple outcomes and incorporate prior distributions for missing correlations. However, a simulation study identifies that external evidence or clinical guidance is needed to ensure appropriate prior distributions for between-study variances and correlations to avoid misleading results. Researchers should thus consider Bayesian methods for clinical trials, but recognise potential difficulties when adopting the approach in practice.

This thesis reports a study into aspects of the discourse concerning the measles, mumps and rubella (MMR) triple vaccine. The aim of the thesis is to contribute to knowledge about the ways in which debates about science are enacted in the public sphere. The study uses a corpus-assisted discourse studies (CADS) approach to examine key lexico-grammatical patterns in the JABS corpus, a corpus of texts gathered from the website of the vaccine-critical Justice Awareness and Basic Support (JABS) group. The aim of the study is to discover how participants on the JABS website discussion forum draw on discursive resources to achieve their rhetorical goals. Comparison is made with the typical lexico-grammatical patterns in the NHSvax corpus, a corpus comprising texts from NHS immunization websites. The study finds that, although there are several areas of similarity between the two corpora, the JABS corpus data contains greater evidence of evaluative lexis, a higher frequency of nouns which express evaluations of the status of discursive objects. These resources are used to reformulate and reframe propositions which originate in the medical-scientific domain. Narratives of vaccine damage are also frequently used to express warrants for expertise.

This thesis consists of two parts. The first part is a systematic review evaluating the available evidence for the use of mindfulness-based interventions (MBIs) in people with neurodegenerative disorders (NDDs). 12 studies met inclusion criteria and involved people with three disorders, Multiple Sclerosis, Parkinson’s Disease and Dementia/Mild Cognitive Impairment. There is promising evidence that MBIs benefit people with a NDD, particularly those with MS in reducing symptoms of anxiety and depression. More high quality studies employing robust methodology such as RCTs with adequate statistical power are needed in PD and dementia/MCI. The second part is an empirical paper focusing on how individuals with acquired brain injury make sense of their experiences of learning and practicing mindfulness skills. Fifteen participants attending a mindfulness group at the local brain injury rehabilitation centre were recruited to two data collection methods. Focus group interviews were analysed using Interpretative Phenomenological Analysis and in-vivo recordings were analysed using Template analysis. Results suggested that most participants considered mindfulness beneficial to cope with emotional and cognitive consequences of ABI. The ‘live’ nature of in-vivo recordings revealed rich descriptions about their individual mindfulness practice. The limitations of this study, future research and clinical implications have been discussed.

Plasmids confer genes encoding clinically relevant antibiotic resistance. It was hypothesised that the AcrAB-TolC multidrug resistance efflux pump was required for clinically relevant levels of carbapenem resistance. However, carbapenemase-producing \(Salmonella\) TolC mutants were less susceptible to carbapenems. In the presence of the efflux inhibitor phe-arg- β-naphthylamide (PAβN), wildtype bacteria and 36/86 non-replicate clinical isolates of carbapenem-producing Enterobacteriaceae were ≥4-fold less susceptible to ertapenem. Experimental data suggested that OmpF repression conferred the increased carbapenem MICs. Two blaKPC-encoding plasmids have been isolated in the UK; pKpQIL-UK was found in K. \(pneumoniae\), but its variant, pKpQIL-D2 was also found in other species. Therefore, it was hypothesised that a region of pKpQIL-D2 either conferred a broader plasmid host range and/or a fitness benefit to the host bacterium. Fitness studies measuring growth rates, ability to form biofilm, conjugation frequency and plasmid persistence showed that both plasmids affected the host bacterium but in different ways. Compared to pKpQIL-UK, pKpQIL-D2 did not confer a significant fitness advantage to its host under the conditions tested. RNAsequencing showed both plasmids affected a different set of genes related to metabolism. The understanding of the factor(s) contributing to the persistence and dissemination of successful plasmids may help to control antibiotic resistance.

Neisseria meningitides is a major cause of meningitis epidemics in sub-Saharan Africa. A promising broadly-protective vaccine approach is outer membrane particle-based vaccine comprising GMMA (Generalised Modules for Membrane Antigens). To better understand vaccine potential and immune responses induced by GMMA, we (i) typed outer membrane vaccine antigens fHbp, NadA, NHBA and PorA in African serogroup A, W and X isolates, (ii) produced GMMA from recombinant strains expressing fHbp variant 1 or NadA-3, and (iii) generated and characterised monoclonal antibodies (mAb) against GMMA. Characterisation of 94 isolates showed limited variability of fHbp, NadA and PorA over time and geographical region. Mice immunised with GMMA developed cross-reactive bactericidal antibody responses against diverse serogroup A, W and X isolates that targeted fHbp, NadA and other unidentified antigens. We obtained 33 hybridoma clones producing GMMA-binding mAbs. Four mAbs of immunoglobulin subclasses IgG1, IgG2a/b and IgG3 bound NadA and four IgG1 mAbs bound fHbp. The other 25 mAbs recognised seven other antigens. In bactericidal assays all anti-NadA mAbs killed the vaccine serogroup A strain. Five mAbs against unknown antigens also showed bactericidal activity against another A isolate. Conclusively, the results provide support for developing a GMMA-based vaccine with broad coverage against meningococcal meningitis for Africa.