This thesis provides a comprehensive analysis of spiritual healing in England in its various different guises during the late-nineteenth and early- to mid-twentieth centuries. It considers the interplay between the various spiritual healing groups themselves and between their philosophies and practices and orthodox medical theory more generally. The first half examines how spiritual healing was conceptualised by those who practised it - who spiritual healers were, what they believed and how they defined illness and healing. The specific therapeutic techniques used by healers are delineated, and the themes of touch and morality explored in detail. The second half of this thesis then examines how spiritual healing was perceived by the religious and medical establishments, and explores their co-operational discourse. Firstly, the reaction of the orthodox Christian churches to spiritual healing and their fractured and inherently conservative attempts to utilise it as a means of revitalising orthodox Christianity are analysed. The final chapters then chart the chronological relationship between spiritual healing and orthodox medicine during three specific periods, and explore the way in which spiritual healing intersected and impacted upon medical reactions to the new psychology of the twentieth century.

This study presents an investigation of the underlying linguistic profiles often Hyperlexic children and explores the nature of the problems which give rise to their diagnosis. The subjects' unexpected exceptional decoding strength together with their similarly unusual reading comprehension failure form the focus of this study. Reasons accounting for both these phenomena are explored. Diagnosis of these subjects is considered in relation to previous definitions of Hyperlexia and claims about its symptoms, nature and association with other deficits. An overview of the controversy and conceptual confusion regarding explanations of Hyperlexia is emphasized. The sources of the Hyperlexic symptoms observed in the subjects are explored and discussed in relation to current psycho-linguistic models of reading and its development. This inquiry leads to two sets of investigations, the first focusing on the subjects' decoding skills and the second on their comprehension and inferencing abilities. The investigation explores a number of questions regarding the subjects' reading skills. These include determining whether the Hyperlexic subjects prefer one route to reading over another (use lexical or sublexical strategies), whether the deficit is modality specific, whether their unusual reading pattern is consistent over time, whether the subjects can access the semantic system and understand words they read as well as the manner in which they approach the learning of novel words (whether semantic cues help or hinder the learning of new words). Findings from the first set of questions leads to a further investigation of the subjects' comprehension failure. Word, sentence and paragraph level semantic and syntactic skills are explored and ruled out as primary sources of the comprehension breakdown. Instead, pr~gmatic language weaknesses are confirmed and a relationship is established between these symptoms and the comprehension failure. The notions of Relevance, Theory of Mind and Central Coherence are discussed and their application to Hyperlexia considered. The concluding discussion addresses a number of theoretical questions regarding the nature of Hyperlexia. Implications for intervention and possible future directions for research are proposed.

Introduction: Opioids are widely used for the management of cancer and chronic non-cancer pain and the maintenance management of patients with a history of substance misuse. Increasingly the use of opioids is being scrutinised as patients are prescribed opioids for longer periods and the long-term effects of the opioids becomes clinically more relevant and evident. Our work has explored the prevalence of opioid-related side-effects in patients who are prescribed opioids and explored the clinically relevant phenomenon of opioid-induced hyperalgesia. . Methods: Patients were recruited who were prescribed opioids for the management of cancer and non-cancer pain or substance misuse. Quantitative data was collected to explore the prevalence and severity of opioid related side-effects, the impact of opioids on cognitive function and the effect of opioids on peripheral nerve function through quantitative sensory testing. Testing the sensory processing of patients who are on opioids has revealed altered thermal thresholds and the presence of wind-up at non-painful sites indicating central sensitisation. Qualitative description was used to explore the patient experience of an episode of opioid toxicity. Results: Patients have a significant burden of side-effects which have often not been recognised by clinicians. Using the Addenbrooke’s Cognitive Examination much more cognitive impairment has been revealed than has previously been recognised. Altered thermal thresholds and wind-up at non-painful sites suggests altered pain processing as a result of opioids. Themes from the qualitative description highlighted the coping strategies patients’ develop when managing with significant side-effects and toxicity, the covert self-management of their pain and the need to exert control. One of the most significant findings from the qualitative research was the finding of altered sensation and pain description associated with other features of opioid toxicity. Conclusions: The impact of opioids on the cognitive function of patients has significant implications in terms of patients’ involvement in decision-making and functioning in everyday life. The qualitative data reflects the burden of side effects and the descriptions of patients suggest that opioid-induced hyperalgesia exists as part of the spectrum of opioid toxicity. This finding may help physicians identify patients who are developing opioid-induced hyperalgesia and allow them to intervene earlier with a proactive approach.

RNA Polymerase III (Poll III) contributes to about 10% of nuclear transcription and is essential for the synthesis of short untranslated transcripts, including tRNA and 5S rRNA. Pol III deregulation has been implicated in driving cellular proliferation and transformation, along with increased expression of a number of Pol III specific transcription factors and transcripts. The significance of Pol III in clinical pathology, including that of human malignancies, remains to be formally tested. Using prostate cancer as a model, two key components of the Pol III complex, namely Brf1 and tRNAiMet, were investigated. The expression patterns of Brf1 and tRNAiMet were studied in this thesis using immunohistochemistry (IHC) and in situ hybridisation (ISH) respectively. Brf1, a subunit of transcription factor IIIB (TFIIIB), was detected predominantly in the nucleus with heterogenous staining intensity, its presence ranging from weak to strong. Examination across a wide range of tissue types and organs, both normal and tumour tissues revealed high levels of Brf1 expression in the epithelium. In addition, Brf1 expression could also be detected in connective tissues and, to a lesser extent, in muscular and nervous tissues. A number of tumour types exhibited elevated expression of Brf1 protein relative to their normal control tissues. These included prostate adenocarcinoma, B-cell lymphoma and, interestingly, tumours arising from connective tissues (sarcoma, fibrosarcoma and chondrosarcoma). As expected, tRNAiMet expression, as revealed by ISH analysis, was mostly observed in the cytoplasm, although some nuclear staining was also present. Similar to Brf1, tRNAiMet expression was detected predominantly in epithelial tissues such as the skin epidermis, prostate gland and epithelial lining of the cervix. A number of tumours were found to overexpress tRNAiMet. These included breast ductal carcinoma, oesophageal carcinoma and melanoma. The clinical impact of Brf1 and tRNAiMet overexpression was examined using tissue microarrays (TMA) containing tissue samples obtained from patients with prostate cancer (PCa) and benign prostate hyperplasia (BPH). Collectively, data from two independent patient cohorts, Glasgow TMA: BPH (n=21), PCa (n=151) and Newcastle TMA: BPH (n=113), PCa (n=365), showed that Brf1 expression was upregulated in prostate cancer relative to BPH (p=0.0034). Brf1 expression was not found to be associated with the following clinical and biologic parameters: Gleason sum score (indicative of tumour differentiation and morphology, p=0.653); prostate specific antigen (PSA level, indicative of tumour bulk or volume, p=0.381) and Ki-67 expression (signifying cellular proliferation, p=0.034). However, and importantly, within the prostate cancer patient cohorts studied, high Brf1 expression was associated with a significantly less favourable survival outcome (Kaplan Meier analysis, p<0.001). Together, the data presented in this study support the relevance of Brf1 and tRNAiMet overexpression as part of Pol III deregulation in tumours, especially of epithelial origin. This study also suggests the potential application of Brf1 as a prognostic marker in cancer, however, this warrants a further study.

I investigated the impact of DARC expression on haematopoiesis using DARC-deficient (DD) mice and developed humanised transgenic DARC models expressing the \(FYB(ES)\) and \(FYB\) human genes. I developed murine irradiation chimeras with differential DARC expression and showed that the absence of erythroid DARC but continued DARC endothelial expression was associated with reduced peripheral blood neutrophil counts. FYB(ES)TG mice showed reduced peripheral neutrophil counts and expansion of BM myelopoiesis, and reduced lymphopoiesis and erythropoiesis, compared to FYBTG mice. FYB(ES)TG and DD mice had reduced GMP cells and LSK cells, and proliferation and apoptosis of these cells were reduced. Microarray analysis of differentially expressed genes showed increased myeloid gene expression in GMP and LSK cells. Mixed irradiation chimeras showed that DD BM retained these changes, suggesting an intrinsic cell effect. Analysis of BM and serum showed a significant increase in G-CSF and eotaxin. Morphological analysis of myeloid cells in femur sections revealed increased myeloid cell clustering in DD and FYB(ES)TG mice. These data suggest that neutrophils are preferentially retained in the DD BM, possibly as a result of loss of optimal chemokine gradients created by erythroblast DARC which may favour neutrophil egress and thereby leading to peripheral neutropenia.

Background: The non-invasive assessment of volume status in left ventricular systolic dysfunction (LVSD) is challenging. The main thesis objective was to establish the feasibility and potential clinical utility of repeated measures assessment of non-invasive biomarkers in defining changes in volume status within individual volunteers. Methods: Differential volume manipulation protocols were achieved in a three-staged plan of investigation, firstly, in patients with decompensated heart failure receiving intravenous furosemide, secondly, in normal volunteers receiving acute loads of oral water or intravenous saline, and thirdly, in stable LVSD patients on chronic furosemide dosing undergoing staged diuretic withdrawal and resumption. Repeated measures of biomarkers relevant to volume status including blood and urine biomarkers, echocardiographic and bioimepdance measures were performed to assess their sensitivity to the induced volume changes. Results Summary: I demonstrated the smallest variance for bioimpedance measures, and the largest variance for urine biomarkers. In the patients with decompensated heart failure, none of the biomarkers studied showed potential clinical utility at tracking acute volume response to intravenous furosemide. In the normal volunteers, a significant change in estimated blood volume was observed following intravenous saline, but not with acute oral water ingestion. Only whole-body and trunk bioimpedance measures, and to a lesser extent, mitral valve early peak velocity with and without the Valsalva manoeuvre, appeared sensitive enough to map these changes in volume status. In the stable LVSD patients, statistically significant increases in B-type natriuretic peptide, urinary creatinine, urinary kidney injury molecule 1, and in bioimpedance-estimated body water composition were observed with diuretic withdrawal, with levels of these markers reducing following diuretic resumption. However, the amplitude of the changes observed was either lower than the respective within-subject variance or too small to be potentially useful as a marker of volume change. This would thus limit the clinical utility of these biomarkers in the routine monitoring of volume status in LVSD. Conclusion: The repeated measures of biomarkers studied in response to different volume manipulations were interpreted in the context of their within-subject normal variance. None of the biomarkers studied appeared to have the ideal characteristics clinically for the monitoring of subclinical changes in volume status in stable LVSD or in response to acute diuresis in decompensated heart failure. The significant increases in urine biomarkers following diuretic withdrawal in stable LVSD suggested potentially beneficial renal effects of furosemide in stable LVSD.

his study investigated the underlying mechanisms of outer membrane homeostasis in Gram-negative bacteria. Using both the evolved laboratory strain E. coli K12 and the broad host range pathogen Salmonella enterica serovar Typhimurium, we have identified and characterised a series of non-essential genes responsible for the maintenance of the outer membrane barrier function. We have revealed their importance for bacterial pathogenesis suggesting their use as novel targets for drug development. This study has provided the first description of a pathway for phospholipid transport from the inner membrane to the outer membrane via the lipoprotein PlpA, a gene previously of unknown function. As several of the genes highlighted by our initial studies were associated with the biogenesis of virulence factors, we complemented our investigations by characterising the contributions of the S. Typhimurium Type V proteins to virulence in a murine model. These investigations have provided the first peer reviewed characterisation of a trimeric autotransporter from Salmonella, has identified a mechanism by which Salmonella can survive on tomatoes, and has highlighted the functional redundancy of these proteins in Salmonella infection. These findings have significantly advanced our understanding of the mechanisms mediating outer membrane homeostasis and the biogenesis and functions of virulence factors.

Eisenmenger syndrome is characterised by severe, lifelong hypoxaemia and pulmonary hypertension. Despite this, patients do surprisingly well and report a reasonable quality of life. This thesis describes a series of experiments investigating the adaptations that occur in these patients in response to the chronic hypoxaemia. Patients with Eisenmenger syndrome have severely limited exercise tolerance when assessed using cardiopulmonary exercise testing. However, they appear to maintain aerobic metabolism until late on in exercise. Studies using skeletal muscle 31P MRS during and throughout recovery from exercise showed that these patients have similar mitochondrial oxidative capacity compared to healthy controls. Echocardiography showed that patients with Eisenmenger syndrome have preserved right and left ventricular systolic function. However they have evidence of right ventricular diastolic dysfunction as evidenced by impaired early diastolic relaxation. The cardiac 31P MRS study demonstrated that despite the normal systolic function shown on echocardiography, there is impairment of septal energetics as revealed by a reduction in PCr/ATP ratio. The results presented in this thesis indicate that adult patients with Eisenmenger syndrome have undergone beneficial adaptations to the severe hypoxaemia that they are exposed to from infancy.

Oral keratinocyte behaviour were analysed in two and three dimensional cultures of an immortalised human H400 cellline and primary rat keratinocytes (PRKs) using a novel method of quantitative microscopy, RT-PCR data and immunohistochemistry profiles. Monolayer cultures were established in high and low calcium media at different cell densities and analysed prior to generating 3D organotypic cultures (OCs) onde-epidermalised dermis (DED), polyethylene terephthalate porous membrane (PET) and collagen gels for up to 14 days.H400 and PRKs proliferation in monolayer cultures was greater in low calcium medium compared with high calcium medium.Gene expression analysis indicated that adhesion and structural molecules including E-cadherin, plakophilin, desmocollin-3, desmogleins-3 and cytokeratins-1, -5, -6, -10, -13 were up-regulated by days 6 and 8 compared with day 4in high calcium medium. Immunohistochemical profiles and gene expression data of OCs on DED recapitulated those of normal oral epithelium. The final thickness of OCs as well as the degree of maturation/stratification was significantly greater on DED compared with other scaffolds used. Quantitative microscopy approaches enabled unbiased architectural characterisation of OCs and the ability to relate stratified organotypic epithelial structures to the normal oral mucosa. H400 and PRK OCs on DED at the air liquid interface demonstrated similar characteristics in terms of gene expression and protein distribution to the normal tissue architecture.

The use of haemostatic materials that could be used to mitigate against the effects of the chemical warfare agent soman (GD) on contaminated personnel that may also present with wounds were investigated. To support the in vitro diffusion cell component of this work, the penetration rate of \(^1\)\(^4\)C-GD into different receptor fluids was evaluated to enable determination of the most appropriate receptor fluid to use as a sink for GD. Of the receptor media evaluated only 50% aqueous ethanol was able to maintain sink conditions. A number of haemostatic materials were shown to retain haemostatic efficacy in the presence of blood contaminated with GD, and were also shown to irreversibly sequester GD. The lead candidate, WoundStat™, was shown to be as effective a decontaminant as the current in service countermeasure fullers’ earth. Complementary in vivo studies using damaged ear skin in a terminally anaesthetised large white pig model showed that whilst use of WoundStat™ was not 100% effective in the prevention of mortality after GD poisoning, it did increase the therapeutic window where further nerve agent-specific medical countermeasures could be employed. Perhaps most importantly, application of WoundStat™ onto GD contaminated damaged skin did not increase the toxicity of GD.