This research reports the study conducted in Saudi Arabia, which aimed to explore: 1) the perception of the term CP with CP children and their mothers, 2) the implication of the meaning for mothers of children with CP, 3) the experiences of being a child with CP in Saudi Arabia. A qualitative approach was taken using critical ethnography methodology, informed by Bronfenbrenner's (1989, 1992 & 2005) ecological theory framework. Data were collected in two phases: Phase one involved two focus group interviews with 6 mothers of children with CP and follow up semi-structured interviews with three of them. The second phase of the research focused on interviews with children with CP. Semi-structured interviews were conducted with 9 children ranging between 5-17 years of age. Talking Mats, an innovative communication tool, was used to enable two children with difficult verbal communication to give their views.

The development of age appropriate paediatric formulations, particularly those suitable for young children, presents challenges with only limited knowledge available on the acceptability of different medicines and how this affects medication adherence. This thesis describes studies conducted at Alder Hey Children’s Hospital, Liverpool UK, with the aim of determining which factors relating to dose form and organoleptic properties of a medicinal product influence medication adherence in chronically ill children. The research was conducted in two phases comprising 70 chronically ill children aged between 3 and 11 years, 70 primary caregivers, and 33 hospital clinical and technical staff.

Pancreatic ductal adenocarcinoma is one of the most aggressive human malignancies, with a five year survival rate of less than five percent. New targets and more effective therapeutic intervention are required to improve diagnosis and prognosis for all patients with pancreatic cancer. Recent studies have established that the tumour suppressor protein Programmed Cell Death Gene 4 (PDCD4) plays a key role in the control of differentiation and neogenesis within the healthy adult pancreas. Pdcd4 was originally identified as a gene up-regulated during the process of apoptosis. However, recent data suggests a function for Pdcd4 as a tumour suppressor, making it a promising target for pancreatic cancer therapy. The present study utilised a novel model of tissue hypoxia, mimicking the oxygen-deprived core of cancerous tumours which is often resistant to conventional chemotherapeutic drugs and which is likely to lead to secondary tumour formation or metastases, especially in pancreatic cancer. Understanding how Pdcd4 regulates pancreatic cell fate, both in healthy pancreatic cells and in conditions of tissue hypoxia, may well arm us with a new weapon in our fight for more effective therapeutic intervention in the treatment and prevention of pancreatic cancer.

Mucus overproduction is an important feature of chronic inflammatory airway diseases including cystic fibrosis, chronic bronchitis, bronchiectasis and severe asthma. Inhaled drugs are often delivered to such diseased airways where mucus hypersecretion is likely to alter the deposition pattern of the drug particles, encouraging more central deposition where more mucus is present than in the airways of the periphery. The effect of the mucus on drug absorption is poorly understood, since there are few in vitro systems capable of modeling the effect of mucus on drug permeability. This project aims to characterize and develop selected lung cell lines as potential models of the airways to study drug permeability in the presence of mucus, whilst also using these models to study potential adverse effects of drug treatment in the airways. Currently there is no published work on drug permeability studies in the presence of mucus, using the chosen lung cell lines.

Synthetic and naturally occurring calcium and magnesium carbonate minerals are widely used in a range of industrial and environmental applications where the mineral quality and purity is often critical for their intended use. Hence accurate characterisation of the mineral assemblages is essential. Equally important is an understanding of the chemical and physical pathways leading to mineral formation and their roles in carbon sequestration from greenhouse gases. This study investigates the application of Raman and infrared spectroscopies to Ca-Mg carbonate analysis. A full quantitative calibration has been achieved for quaternary mixtures by Raman spectroscopy (RS) employing monovariable and multivariable methods. The method was validated by X-ray powder diffraction (XRD). The lowest error on component values was obtained by Principal Component Regression with application of Standard Normal Variate. The quantifications show that RS is comparable to XRD. The effect of particle size on the fundamental vibrations of the [CO32-] anion in calcite is investigated by mid-infrared and RS. While the effect of particle size on the infrared signature of internal modes of the [CO32-] anion is well documented, this thesis documents associated changes in Raman spectra as a function of particle size. With decreasing size spectral contrast diminishes and changes in the relative ratios of the internal modes occur. For RS the turnaround from optically thick to thin material occurs in the 42-59 μm size range with further changes occurring at ≤ 5 μm. RS was also utilized to monitor carbonate reaction kinetics after dissolution of [Mg(OH)2] by CO2 sparging in the presence of calcium salts at 35 °C, 30 days duration. Four experiments employing different calcium salts, Ca:Mg ratios and effect of hydromagnesite [Mg5(CO3)4(OH)2.xH2O] seeding were examined utilizing vibrational spectroscopies, XRD and SEM. Results suggest that carbonate mineral paragenesis is driven by geochemical feedback between a range of calcium and magnesium carbonate dissolution-precipitation events where decomposition of nesquehonite [Mg(HCO3,OH)∙2H2O] leads to formation of magnesium carbonate hydrates [Mg5(CO3)4(OH)2.xH2O]. XRD confirmed that these hydrated phases contain 8 and/or 5 molecules of crystalline water. However, RS cannot distinguish these phases. Traces of barringtonite [Mg(CO3)∙2H2O] found at the end of experiments were interpreted as an indicator of incongruent dissolution of nesquehonite. Findings suggest that the Raman active ν1 mode of barringtonite is situated at ca. 1094-1095 cm-1. The limitations of Raman analysis in the context of mineral assemblage quantification, short range ordering and particle size effects are discussed in the context of these findings.

Resveratrol and compounds related to it, known as resveralogues, have been shown to extend healthy lifespan in a number of species. A diverse set of molecular mechanisms have been proposed to account for these effects, including the activation of the NAD+-dependent histone deacetylase SIRT1. However, both resveratrol and some of its derivatives also display potentially detrimental activities including direct DNA damage, toxicity and the induction of cellular senescence. Accordingly, the overall aim of this thesis was to enable the generation of better compounds with minimal detrimental activity and maximum beneficial activity. The data presented in this thesis show my development of a new, convenient, one-pot stereo-selective synthesis of resveratrol and other trans-stilbene derivatives. This synthesis has been used to prepare more than 40 compounds, of which 20 are completely novel and a further 12 previously uncharacterised with regard to their biological effects. This study have evaluated these resveralogues with respect to their cytotoxicity in human fibroblasts using a range of widely-recognised assays. Data on the compounds’ toxicity are presented, together with evidence that two of the assays commonly used to measure cytotoxicity in vitro (the MTT and neutral red assays) are unsuited for cytotoxicity testing with this class of compound. Twenty-two of the (lowest toxicity) compounds have also been assayed for their effects on growth and senescence using indirect immunostaining for the proliferation marker pki-67 and catalytic histochemical visualisation of SA-β-galactosidase activity, respectively. The results indicate that the growth inhibitory activity of resveratrol is abrogated by the removal of the trans-stilbene double bond (no reduction of growth fraction in dihydroresveralogues at 100 μM). Interestingly, at low doses (5-10 μM), many resveralogues induce a significant increase in proliferation compared to control untreated cells. SIRT1 activity in the presence of each compound was measured using an in-vitro deacetylation assay and, whilst most resveralogues are SIRT1 activators, none of the compounds examined produced a significant increase in SIRT1 activation compared to the parent compound. SIRT1 activation was, for the first time, found to be independent of the presence of a trans-stilbene double bond. Some initial characterisation of the ability of the resveralogues to alter pro-inflammatory cytokine expression was also undertaken by using ELISA for interleukin 6. Some difficulties with this assay, with respect to background signal interference, were encountered but parallel data from collaborator (Professor Lynne Cox, University of Oxford) using a human specific IL6 antibody are available. Collabrator data, evaluating a selection of resveralogues, indicate that inhibition of IL6 release is independent of SIRT1 activation, demonstrating that not all potential beneficial effects are SIRT1 mediated. Finally, this work has identified three novel lead compounds (V24, V31, and V34) that have less detrimental activities with retention in SIRT1 activity.

An important aspect of health visitors’ (HVs’) role and responsibility is to identify and respond to domestic violence and abuse (DVA). To date, there has been limited exploration of HVs’ practice knowledge and the nature of their professional relationships during their day-to-day practice in addressing incidents of DVA in black and minority ethnic (BME) communities. Aim - The aim of the study was to explore the nature of HVs’ practice knowledge and professional encounters when trying to define, identify and respond to incidents of DVA in BME communities. Design - This study adopts the interpretative lens of a postcolonial feminist theoretical perspective as the focus for analysing the nature of HVs’ knowledge and their practices in addressing issues of DVA in BME communities. Postcolonial feminist thinkers offer the conjecture that knowledge about BME women’s lives must be analysed within the intersecting racialised, gendered and political contexts of their lives. The study utilises a mixed-method approach by conducting semi-structured interviews with twenty health visitors (HV) and documentary analysis of four key professional practice guidance documents on addressing DVA. Findings - The findings revealed the extent to which HVs’ theoretical and personal knowledge and practice in addressing DVA in BME communities are informed by the racialised, familial, gendered and political settings in which they work. The findings illuminate the complexities that are shaped by the neoliberalist approach to tackling health inequalities in the modern National Health Service (NHS). In particular, the study conceptualise the nature of HVs’ professional relationships with BME women to uncover DVA as a form of hegemonic representation. The findings offer the potential to transform the education and practice of current and future health professionals for the benefit of BME and other marginalised patients or service users. Implication for Practice - This research recommends a practice model which seeks to prioritise emancipatory knowledge. In particular, there are recommendations on the specific context in which HVs work with BME women to uncover DVA. It is suggested that further research in this area of practice should also explore the impact of a proposed Intersectionality framework for uncovering DVA in marginalised groups. This study represents an original contribution to knowledge by increasing understanding of the ways in which HVs work to address DVA in BME communities. Although implied in the literature, the understanding of HVs’ work at the intersection of DVA, familial and political perspectives have never previously been articulated in HV literature in this way.

Low back pain (LBP) is the most common form of chronic pain. Approximately 85% to 90% of chronic low back pain (CLBP) episodes in primary care cannot be related to serious pathology or neurocompression, being described as nonspecific chronic low back pain (NSCLBP). This disorder involves continuous pain or recurrent flare-ups that are responsible for high levels of distress, functional disability and work absenteeism. It has also a significant impact on health care systems and society in general. Considering the complexity inherent in the experience of NSCLBP, where personal and cultural contexts play a major role, research has highlighted the need to study this phenomenon in contexts that have not yet been investigated. This study aimed to explore the Portuguese individuals` experiences and perceptions of NSCLBP. An interpretative phenomenological analysis (IPA) was employed to explore the experiences of eight participants, who were recruited purposefully from three Portuguese health sites. Semi-structured one-to-one interviews were carried out in order to collect data. The interviews were audiorecorded and transcribed verbatim. Following an inductive process of data analysis, five themes emerged as interrelated parts of an extended account that explored the Portuguese individuals` experiences and perceptions of NSCLBP. In the first theme the disruptive nature of the NSCLBP experience was emphasised. In the second, the participants` meaning making of NSCLBP and their need to understand it were highlighted. In the third, the clinical encounters and their contribution to maintaining the lack of participants’ understanding about NSCLBP were emphasised. In the fourth, the meaning making of NSCLBP contribution to reshaping the participants` social interactions was explored. In the fifth, the participants` definition of their sense of self through the meaning making of NSCLBP was highlighted. In order to promote the transparency of data analysis, an audit trail was developed to document all relevant steps of this process. This study has offered the first insights into the Portuguese individuals’ experiences of NSCLBP disorder, which may help clinicians in transferring this knowledge to the therapeutic approach to patients with similar experiences. The knowledge produced may be used to inform recommendations for NSCLBP management.

Diabetes is currently a wide spread global disease. As a result of the side effects of the current therapies, herbal plants may present alternative source of drugs for management of the disease. Mucuna pruriens is a plant that is traditionally used for diabetes and anaemia. There are experimental reports of the hypoglycaemic effect of the alcoholic extracts but the anti-diabetic effects of the aqueous extract has not been investigated. Therefore, the aim of this project was to investigate the potential anti-diabetic mechanisms of the aqueous extract of Mucuna pruriens leaves. The leaf extract was prepared by decoction. The potential mechanisms of anti-diabetic effect of this extract was evaluated as follows: Antioxidant activity of the aqueous Mucuna pruriens leaf extract was investigated in reduced β-nicotinamide adenine dinucleotide (NADH)/phenazine methosulphate (PMS), and Xanthine /Xanthine oxidase superoxide generating systems. In addition, the effect of aqueous Mucuna pruriens leaf extract against oxidative stress was measured as cytoprotective effect of the extract against paraquat induced oxidant injury in NRK-52E renal cells. Cytoprotective effect was measured as cell viability and cell death using 3-(4,5-Dimethylthiazol-2-Yl)-2,5 Diphenyltetrazolium Bromide (MTT) and Lactose dehydrogenase (LDH) assays respectively. Finally the effect of aqueous Mucuna pruriens leaf extract on glucose uptake was evaluated in NRK-52E renal cells and 3T3-L1 adipocytes. The results revealed that aqueous Mucuna pruriens leaf extract had significant superoxide scavenging activity which increased from 21.35% to 99.8% in xanthine/xanthine oxidase and 36.15% to 62.4% in NADH/PMS superoxide generating systems at p < 0.05. However, aqueous Mucuna pruriens leaf extract did not protect against paraquat induced oxidative stress. Data from glucose uptake experiments showed that 1mg/ml of aqueous Mucuna pruriens leaf extract inhibited glucose uptake in NRK-52E renal by 35.5% compared to control at p < 0.05. This effect was comparable to 1mM Phloridzin (a non- selective inhibitor of sodium glucose transporters). Finally, 50 and 100μg/ml of both aqueous Mucuna pruriens leaf extract and its acid hydrolysed fractions prepared with liquid-liquid partitioning in diethyl ether, stimulated glucose uptake in 3T3-L1 adipocytes. Specifically, 50 and 100μg/ml aqueous Mucuna pruriens leaf extract stimulated glucose uptake be 57.06 and 86.24% respectively compared to negative control at p < 0.05. Increase in glucose uptake was also observed in cells treated with diethyl ether acid hydrolysed fractions. Taken together, the results show that aqueous the Mucuna pruriens leaf extract used in this study may exert anti-diabetic effects via antioxidant and glucose uptake modulatory mechanisms.

Prokaryotic toxin-antitoxin (TA) systems (also known as addiction modules), are ubiquitous genetic modules first discovered due to their role in stabilising vertical transmission of plasmids. Generally they are two-gene systems encoding a stable toxin (Tx) and an unstable antitoxin (ATx). Loss of the TA module leads to rapid ATx degradation and depletion, leaving the Tx free to interact with cellular targets and inhibit growth. For plasmid encoded TA systems, this leads to the death of plasmid free daughter cells, ensuring plasmid maintenance in a population, and gives rise to the term "addiction module". More recently, the expansion in microbial genome data has highlighted the prevalence and diversity of TA systems, and demonstrated that they are common features of many bacterial chromosomes. In addition, metagenomic surveys have pointed to the enrichment of some TA families in particular microbial ecosystems; a prime example from surveys of the human gut microbiome and RelBE TA family. Collectively, these observations indicate a wider role for TA modules in bacterial function, with numerous roles for TA systems now hypothesised. These include: i) Stabilisation of TA associated chromosomal DNA during vertical transmission; ii) Formation of "persister" cells resistant to environmental stresses, and; iii) Population level resistance to bacteriophage attack. Additionally, some Tx components have shown activity in eukaryotic cells, raising the potential for a role in prokaryote-eukaryote interaction. Here we undertook a systematic study of Type II TA systems, to provide a comprehensive assessment of their distribution and relative abundance, to confirm activity of prevalent TA systems, and to understand putative roles these may play in gut associated bacteria and the gut microbiome. A comparative genomic and metagenomic analysis of 3919 bacterial chromosomes, 4580 plasmids, 711 bacteriophage genomes, and 781 metagenomes encompassing 16 distinct habitats was conducted using all known Type II TA systems present in the Toxin Antitoxin Database (~10,100 TA genes ~1:1 Tx:ATx). Of the 817 Type II TA system homologues found in human gut datasets, 686 were observed to have significantly higher relative abundance in the human gut microbiome over other microbial ecosystems. In parallel to these in silico findings, PCR and qPCR surveys of microbiomes from 65 stool samples obtained from healthy volunteers, as well as those with polyps or colorectal cancer, were undertaken. This demonstrated a higher ATx presence than Tx or complete module, however no differences in Tx copy number between health groups was seen. To confirm the activity of the most abundant TA system homologues identified in sequence surveys, ORFs were amplified from gut metagenomic DNA, and individual Tx or ATx cloned under the control of inducible promoters. Induction of Tx expression under normal growth conditions resulted in bacterial growth inhibition, while live dead staining showed entry into a viable but non-cultivatable state, commensurate with TA function. Experiments simulating environmental stresses encountered during colonisation of the GI tract (starvation, low pH, bile), indicated that expression of these TA systems could increase cell survival when carbon or nitrogen availability was limited (starvation). Since antibiotics are also commonly encountered by gut associated-bacteria (both as residents of the GI tract and during colonisation of other body sties) a role for gut associated TA systems in facilitating survival during antibiotic exposure was also explored. This revealed an increased number of cells surviving two hours post-treatment with β-lactams when Tx genes were expressed, and in keeping with an impact on cell growth. To test the hypothesis that TA systems may stabilize associated regions of DNA, the composition of gene neighbourhoods surrounding TA systems were also explored. ORFs surrounding TA system homologues identified in metagenomic and genomic datasets were identified using the Metagene annotator, and ORF functions predicted based on searches of the Clusters of Orthlogous Groups (COG) database. This revealed significant increases in ORFs with functions related to replication/recombination/repair and those with unknown functions. It also identified a decrease in the proportion of ORFs encoding functions such as carbohydrate and lipid transport and metabolism in regions surrounding TA systems, suggesting involvement with stabilization of mobile elements. Finally, we explored the potential for gut associated TA systems to modulate phage-microbe, and host-microbe interactions. In the case of phage-host interactions, TA systems have previously been shown to function as mediators of phage resistance at the population level, by directing cells towards a dormant state which prevents phage replication, and permits a sub-set of cells to survive phage attack. Our findings indicated the potential for gut associated TA systems to provide some degree of protection during particular host-phage interactions, but specific modules did not provide universal protection against phage. In the case of host-microbe interaction, some Type II TA system Tx components have been shown to be functional in cultured eukaryotic cells, promoting apoptosis when introduced and expressed in these cell types. However, no studies to date have examined the potential for bacterially expressed TA systems to influence eukaryotic cell health in co-culture models. To investigate this, we assessed the impact of bacterial TA system expression on the health of the intestinal epithelial cell line Caco-2 in co-culture systems specifically focusing on cell apoptosis and necrosis whilst in the presence of Escherichia coli expressing p22-RelBE.