The presence of chronic inflammation is associated with increased nutrient availability during obesity or type 2 diabetes which contributes to the development of complications such as atherosclerosis, stroke and myocardial infarction. The link between increased nutrient availability and inflammatory response remains poorly understood. The functioning of monocytes, the primary instigators of the inflammatory response was assessed in response to obesity and increased glucose availability. Monocyte microRNA expression was assessed in obese individuals prior to and up to one year after bariatric surgery. A number of microRNAs were identified to be dysregulated in obesity, some of which have previously been linked to the regulation of monocyte inflammatory responses including the microRNAs 146a-5p and 424-5p. Weight loss in response to bariatric surgery lead to the reversal of microRNA changes towards control values. In vitro treatments of THP-1 monocytes with high concentrations of D-glucose resulted in decreased intracellular NAD+:NADH ratio, decreased SIRT1 deacetylase activity and increased P65 acetylation. However the increased osmotic concentration inhibited LPS induced inflammatory response and TNFα mRNA expression. In vitro treatment of primary human monocytes with increased concentrations of D-glucose resulted in increased secretion of a number of inflammatory cytokines and increased expression of TNFα mRNA. Treatment also resulted in decreased intracellular NAD+:NADH ratio and increased binding of acetylated P65 to the TNFα promoter region. In vitro treatments of primary monocytes also replicated the altered expression of the microRNAs 146a-5p and miR-424-5p, as seen in obese individuals. In conclusion a number of changes in monocyte function were observed in response to obesity and treatment with high concentrations of D-glucose. These may lead to the dysregulation of inflammatory responses contributing to the development of co-morbidities.

Cell tracking is important because the information it provides is required to inform cell based treatment development. Additionally, it has a diagnostic purpose for detecting infection/inflammation diseases with labelled cells, a technique that has become part of routine clinical practice. Currently only gamma emitting radiotracers for single photon emission computed tomography (SPECT) are available to label cells for routine service. PET (positron emission tomography) exhibits better resolution than SPECT and its use in cell tracking would improve image quality, detectability and quantification for smaller lesions. The studies presented in this thesis report on development of cell labelling techniques with positron emitting radionuclides. The production and characterisation of complexes of copper-64 with dithiocarbamate (DTC) analogues and bis(thiosemicarbazones) (BTSCs) are described. Their labelling efficacies and stabilities were investigated. Although efficient and rapid extraction of 64Cu (DEDTC)2 and 64Cu-GTSM were observed, a high percentage of the radiotracer leakage from the labelled cells was still problematic. Efflux kinetics were similar for all the complexes used suggesting that the efflux mechanism was the same in all cases. In an alternative approach, a novel lipophilic complex of the long lived PET isotope, zirconium-89, was synthesised and characterised. [89Zr]-Zr(oxinate)4 showed good retention in the labelled cells 24 hr after labelling as well as moderate uptake in many types of cells including human leucocytes and mouse multiple myeloma (MM) cells (eGFP-5T33). In vivo imaging and ex vivo tissues counting was used to compare cell tracking with [89Zr]-Zr(oxinate)4 and [111In]-In(oxinate)3 labelled MM cells. The main organs of the homing radiolabelled cells were similar in MM model. However, mice injected [111In]-In(oxinate)3 labelled cells showed higher accumulation of activity in the kidneys than those of [89Zr]-Zr(oxinate)4 that might indicate the greater release and metabolism of the 111In radiotracer released from the labelled cells. The localisation of [89Zr]-Zr(oxinate)4 in labelled MM cells was confirmed by sorting of homing organ (liver, spleen, bone marrow) homogenates based on green fluorescence protein (GFP) expression up to 7 days post inoculation, which showed that radioactivity remained predominantly in GFP-positive cells confirming that radionuclide loss from the labelled cells was minimal and that the cells remained alive at 7 days post injection. In addition we demonstrated that mice inoculated [89Zr]-Zr(oxinate)4 labelled cells can be tracked by PET imaging for 14 days after inoculation. It was concluded that while Cu-64 radiolabelling of cells was ineffective because the majority of radioactivity was lost from cells by 24 hr, the novel complex [89Zr]-Zr(oxinate)4 can be successfully synthesised with acceptable quality and very promising long lived PET tracer for tracking labelled cells for up to weeks.

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The purpose of this thesis was to examine the relationship between adult attachment style and health. Design Three questionnaire studies and one interview study were completed. Method . Questionnaires were administered to examine the relationship between attachment style and symptom reporting in healthy undergraduates (study 1) and cardiac patients (study 3). In Study 2 physiological response to stress was also measured. Study 4 used a semi structured interview approach to examine illness experience in cardiac patients. Results In study 1 insecurely attached students reported more somatic, anxious, social dysfunction and depressed symptoms than secure students. This relationship was mediated by anger and social support. In study 2 baseline differences in cardiac output and total peripheral differences were found, however, no differences were found in response to stress. Furthermore, insecurely attached students reported more depressed symptoms than those classed as secure. No mediators were identified. In study 3 the relationship between attachment styles on symptom reporting was . examined in chronic heart failure (CHF) and transplant populations. Insecure CHF patients reported a greater number of symptoms than secure patients. Anger and social support mediated this relationship. Only one difference was reported in the transplant group, with insecurely attached transplant patients reported more depressed symptoms than those classed as secure. Finally,interviews in study 4 identified four themes of control, normality, social support and emotional disclosure. Standardised symptom reporting tools may not capture these elements of the patient experience, and attachment style may offer one explanation for the variations reported by respondents. Conclusion In conclusion, the results of this thesis support attachment related differences in symptom reporting behaviour in clinical and non clinical populations, and that anger and social support mediated this relationship. The research suggests that attachment style may be a valuable tool to be utilised within the health services.

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