Technologies such as geographical information systems (GIS) have emerged during the past two decades as part of the Information Revolution and include functions such as data storage, management, integration, analysis and presentation. GIS have wide and diverse applications in disciplines such as engineering, business/marketing, urban planning and environmental management but remain underused in public health. The thesis reports the findings of a mixed methods study examining the views and perceptions of public health practitioners on the value of GIS mapping in decision-making. A case study design was chosen; the case issue (childhood lead [Pb] exposure) represents an example of the "case" which is defined as 'decision- making in public health'. The exploratory phase of the study combined heterogeneous data to produce a visualisation of lead contamination in Newcastle. The value of GIS in public health was explored in an interview phase. Twenty-two semi-structured interviews were conducted with decision-makers involved either directly or indirectly in public health practice. Interview recordings were transcribed and coded thematically for analysis. Decision-makers tended to be positive about the use of GIS in public health and many volunteered potential opportunities to apply GIS mapping techniques further. Four discourses were highlighted through analyses, namely: data origins (Ontological Discourse), status (Power Discourse), application (Functionality Discourse) and reciprocity (Collaboration Discourse). The power of maps to integrate multiple, disparate datasets was found to be important and respondents felt, overall, that GIS mapping was a democratic means of communication. Complexity frameworks are drawn upon to make sense of the research findings and to illuminate the need for non-reductionist models of decision-making in the public health context. The lessons learnt through this study can be translated to other fields, thereby sharing skills, knowledge and experience to promote collaboration and integrated thinking across the public health landscape.

This work is based on eight papers published between 1998 and 2006. The papers present a process of investigating, discussing and documenting how, through exploring, stretching and developing opportunities offered by various qualitative research approaches, facilitated collaborative action research (CAR) and evaluation became entwined They question how and where participants in projects recognise their own knowledge and learning, and how they use and develop their understandings in relation to new knowledge. In these papers I worked at the interface between the known and the nearly known; between knowledge-in-use and tacit knowledge that was yet to be useful. This interface, a 'messy area', was a place of contested knowledge. In this 'messy area' long-held views, shaped by pro essional knowledge, practical judgement, experience and intuition, came together to disturb both individual and communally held notions of knowledge for practice. Working in the 'messy area' enabled new knowing that has both theoretical and practical significance to arise, a 'messy turn' to take place. This is the purpose of mess. These papers add to the body of knowledge about 'seeing' and 'knowing', 'the importance of not knowing' and the role of participation, collaboration, facilitation and learning as key change mechanisms in research and evaluation.

The progressive loss of cartilage matrix is a major characteristic of arthritic disease, ultimately leading to a loss of joint function. A number of therapeutics are used in the treatment of arthritic disease, with non-steroidal anti-inflammatory drugs used to treat the pain and inflammation seen in osteoarthritis and rheumatoid arthritis, whilst disease modifying anti-rheumatic drugs are used to slow disease progression. However it is not fully understood if and how many of these drugs effect the disease processes in arthritis. The objective of this study is to look at a number of therapeutics and investigate their effect on the breakdown of proteoglycan and collagen, and on the expression of a number of key degradation enzymes, whilst trying to identify the possible mechanisms used by the drugs. Using the bovine nasal cartilage explant model IL-1 + OSM were used to stimulate the release of proteoglycan and collagen from the cartilage. Interleukin-1 (IL-1) and oncostatin M (OSM) together promote the degradation of cartilage by up regulating and activating MMPs that are found within the diseased joint. Treatment of the resorbing cartilage with indomethacin, indomethacin heptyl ester, simvastatin, mevastatin, pravastatin and sulfasalazine produced a variation of findings with many resulting in the inhibition of cartilage degradation. The stimulation of human articular chondrocytes with IL-1 + OSM caused a significant up regulation of MMPs by the cells at both a gene and protein level when measured by TaqMan PCR and ELISA respectively. Again treatment of the stimulated chondrocytes with a number of the drugs showed a significant reduction in the expression of key cartilage degrading enzymes. The investigation of signalling mechanisms affected by one specific drug, namely sulfasalazine by immunoblotting and signalling microarray, showed some interesting results not previously documented. Sulfasalazine is well known to inhibit NF-I(13 activation by blocking the degradation of IicB to the proteosome and data in this study supports these finding. However, study of the MAPK pathway showed that sulfasalazine appears to be able to block the signalling cascade which ultimately leads to AP-1 activation in chondrocytes stimulated with IL-1 + OSM. This study has identified possible chondroprotective properties in a number of the drugs which were screened and whilst the exact mechanisms behind these events still require further investigation, the results highlight the potential of these drugs in being used to prevent further cartilage degradation in arthritis and thus further delaying the possible need for joint replacement operations.

Disability has a profound impact on a person's quality of life (QOL). Rehabilitation, a process by which measures are taken to improve the QOL of people with disability (PWD) uses several approaches. Community Based Rehabilitation (CBR) is one such approach, which evolved because of the magnitude of the problems of PWD in the community, the limited availability and poor access of rehabilitation services. Research in CBR is limited and hence there is lack of evidence on outcomes in CBR. Considering the need for more information on current practice and research in CBR, the researcher studied the effectiveness of an integrated model of CBR set up in the Christian Medical College, Vellore, South India (VCBR). The objectives of this study were to generate theory on the value of an `integrated model' that uses an educational strategy and to explore the value of secondary and tertiary care services for PWD in a community based rehabilitation programme for the improvement of their quality of life. A 'Case Study' design was used. The practitioner role of the researcher added an important component to this study. An in depth study of 20 PWD, their immediate family members, concerned trained volunteers (LS) and others who were involved in VCBR was undertaken in addition to observation and reference of documents. Qualitative Analysis was undertaken based on the Framework Technique. The quality of life of PWD and the role of secondary and tertiary care centres in VCBR were studied. In this study the realist approach, which takes note of the contextual elements in the evaluation of case study materials, showed that overall QOL of PWD will not improve to its full potential if solutions are mooted from polarized viewpoints. The study found that an integrated model of CBR that uses an educational strategy, has good links with secondary/tertiary care centres and makes use of social network/capital, which is available in the community, improved the overall QOL of PWD.

Acute lymphoblastic leukaemia (ALL) is the commonest of the childhood cancers, but thankfully responds well to chemotherapeutic agents, with 80% of children achieving long-term survival. However, for the remaining 20% who relapse, outcome is bleak. Increasing knowledge and understanding of pharmacogenetics indicates that constitutive or acquired resistance to the drugs used in the treatment of cancer contribute to relapse. Methotrexate (MTX) is one of the most important drugs used in the treatment of ALL and the work presented here contributes to the body of knowledge relevant to the understanding of resistance to this drug. The aim of this research is to determine if changes in genes involved in folate metabolism contribute to methotrexate resistance and subsequent relapse of childhood ALL. The reduced folate carrier (RFC) is required to transport methotrexate into the cell where it competitively inhibits dihydrofolate reductase (DHFR) and other key enzymes of folate and 1-carbon metabolism. Following the design of primers and optimisation of PCR amplification, the entire coding regions of both RFC and DHFR were screened for novel SNPs or mutations using denaturing high performance liquid chromatography (DHPLC) on genomic DNA from 40 normal and 40 relapse childhood ALL patients. Ethical approval for use of the samples was granted under WCRLEC no347 and reference 2002/111 for normal and relapse samples respectively. The relapse group was made up of 29 males and 11 females with an average age of 6.59 years (range 0.8-14.1) and was made up of 5 T-cell; 30 B-cell; 4 mixed lineage; and 1 null classification. The screening method was shown to be sufficiently sensitive to detect single base changes. Several of the exons in both genes have a high G-C content and required destabilisation agents and/or use of a new DNA polymerase (OptimaseTM) to achieve sufficient PCR amplification. Dimethyl sulphoxide (DMSO) and betaine were shown to be effective agents which did not interfere with subsequent DHPLC. The results show that somatic mutations in the coding sequences of RFC and DHFR are rare in relapsed ALL and that while it is recognised as a mechanism of methotrexate resistance in vitro, it is unlikely to contribute to relapse in children with ALL. However, for one patient who suffered multiple relapses, a novel acquired mutation was identified in the 5'-UTR of the RFC-1 gene (C-37T). The significance of the C-37T mutation on RFC transcription requires further study, but it may decrease RFC mRNA quantity or stability and thus protein levels. DHPLC analysis also detected common SNPs. In terms of frequency, there were no significant differences between relapse and normal samples for the genotypes; RFC G80A (38.3% G/G; 49.4% G/A; 12.3% A/A in 81 normal samples; 31.8%; 56.8%; 11.4% respectively in 44 relapse samples; x2 p = 0.656); RFC C696T (97.8% C/C; 2.2% C/T; 0% T/T in 45 normal samples; 92.5%; 5%; 2.5% respectively in 40 relapse samples; x2 p = 0.458). However, there was a significant difference in the 5'-UTR -19 base pair deletion in the DHFR gene between the normal and relapse groups (0% WT/WT; 100% WT/-19 or -19/-19 in 45 normals; 26.7% and 73.3% respectively in 15 relapse samples; Fisher's exact probability p = 0.017) and needs to be confirmed in a larger cohort. Estimated copy number and the probability of loss of heterozygosity (LOH) were generated from the Affymetrix 50K SNP microarray for the RFC, DHFR and other genes involved in folate metabolism using gDNA from 73 presentation and 20 relapse childhood ALL cases. Mann-Whitney non-parametric comparison of the presentation and relapse data showed that for estimated copy number, statistically significant differences were seen for MTHFR (p = 0.0072), MS (0.0025), FPGS (0.00048), TS (0.00046), CBS (0.0002) and RFC (0.0001). When the data at the nearest SNP location to the gene was presented as a scatterplot, the difference in each case was due to a bimodal distribu...

Bacteroides fragilis NCTC 9343 is a Gram-negative anaerobic bacterium with genomic DNA of 5205 Kb and a GC ratio of 43%. It is a commensal organism that can act as an opportunistic pathogen and is commonly present on the mucous membranes. It causes a variety of infections including intra abdominal infections, perirectal abscesses and decubitus ulcers. Enterotoxigenic forms are capable of causing diarrhoea in children and animals. Enterobacter cancerogenus ATCC 35316 is also a Gram-negative facultatively anaerobic bacterium with genomic DNA of 4602 Kb and a GC ratio of 55%. It is a naturally occurring human gut symbiont known to exhibit resistance to antibiotics like aminopenicillins. It has also been reported in cases of severe osteomyelitis and infections of bones and joints. This study aims to analyse the differential expression of proteins in the presence of mucin since it serves as the first site of adherence for the bacteria. The E. cancerogenus and B. fra gilis proteins were extracted and separated by two dimensional electrophoresis from logarithmic phase cultures grown in semi-defined media enriched with or without porcine gastric mucin Types II and III. The gel images were analysed using Bio-Rad PDQuest, Ludesi Redfin and Nonlinear Dynamics SameSpots softwares. It was observed that the presence of mucin in the media affected the expression of a number of proteins in E. cancerogenus and B. fragilis cells. The protein spots of interest were excised, hydrolysed using trypsin and subjected to electrospray ionisation based LC-MS analysis in order to determine the identity of the digested proteins and obtain a better understanding of the interactions of B. fra gilis and E. cancero genus with mucin. The outer membrane protein surface antigen X was found to be up-regulated in both mucin Type II and III enriched media in E. cancerogenus. Some of the other proteins that were differentially regulated in both E. cancerogenus and B. fra gilis included the elongation factor Ts, malate dehydrogenase, triose phosphate isomerase and thiol peroxidase proteins indicating that these proteins may be associated with the ability of bacteria to grow in mucin and may be potential virulence factors. Genes encoding the proteins CAH06598 and CAH09443 from the glycoside hydrolase families 95 and 97 in B. fra gilis strain NCTC9343 were cloned, overexpressed and purified using nickel affinity and gel filtration chromatography. The enzymes were found to be active by performing fluorimetric assays using methyl-umbelliferyl sugar substrates. Diffracting crystals of CAH09443 were obtained from the PACT ANION screens containing polyethylene glycol and sodium malonate as a precipitant. Structure determination was achieved via molecular replacement using the glycoside hydrolase Family 97 α-galactosidase, BtGH97b, from Bacteroides thetaiotaomicron as a starting model. The structure of CAH09443 was shown to be composed of a N-terminal β-super-sandwich domain and a canonical (β/α)₈ barrel, similar to the two other glycoside hydrolase family 97 enzyme structures reported.

The last fifty years has seen an increase in the production of synthetic or artificial enzyme substrates used to identify and quantify enzymes. These substrates have found applications in a range of biomedical science disciplines. Used in biochemistry and clinical chemistry to identify and measure enzymes, some of these substrates have been adapted for use in microbiology, particularly bacterial diagnosis and, in more recent years, molecular biology. The use of artificial chromogenic and fluorogenic enzyme substrates to identify certain bacteria is now common place in medical laboratories worldwide. Not all bacteria can be identified with existing and commercially available artificial substrates. Some of these can be slow to yield results, imprecise, expensive or require a technical method too complicated to provide a viable laboratory test. Therefore, the search for new, more efficient, biochemical tests has progressed, with novel substrates and inventive applications being developed continually. In this study, core compounds were synthesised by various condensation reactions and their characteristics evaluated with respect to colouration/fluorescence and possible enhancement of these properties by metal chelation. Promising candidates were selected for glycosidation, via modified Koenigs-Knorr reactions, in an attempt to synthesise artificial substrates. Several commercially available core molecules were also subjected to glycosidation. The more successful substrates included glycosides of alizarin, nitrosalicylaldehyde and 3- hydroxyflavone. The galactoside of nitrosalicylaldehyde was evaluated in solid agar media and found to be selective for certain Gram-negative bacteria. When similarly investigated, the 3- hydroxyflavone-β-D-glucoside showed the possibility of being used in a procedure for the isolation of the clinically significant pathogens including Listeria monocytogenes. The enzyme kinetics of β-glucosidase with this substrate were also determined in a novel fluorescence assay and compared favourably to the well documented 4-methylumbelliferyl-β-D-glucopyranoside. Alizarin-2-yl-β-D-galactoside and p-naphtholbenzein-β-D-galactoside were successfully utilized for the screening of recombinant and non-recombinant Escherichia coli transformants produced routinely in molecular biology. Aminopeptidase substrates have been shown to be useful for the detection of enzymes which hydrolyse peptides that are specific to certain bacteria. To allow the evaluation of novel aminopeptidase substrates, that were to be subsequently synthesised, a cost effective, large scale source of recombinant leucyl aminopeptidase enzyme was developed via gene cloning techniques. Consequently, the products of this study may serve a beneficial purpose in future enzymatic investigations, medical diagnosis and molecular biology.

No description available.

This study used a qualitative method to focus on the perspectives, beliefs and expectations of low back pain sufferers. The research was undertaken within a hospital based pain clinic. In recent years low back pain research has proliferated, and the epidemiological evidence suggests that back pain is an increasing problem. Much attention has been paid to the impact of low back pain on the population, and to the increasing cost in economic and health terms. Biomedical and psychological evidence abounds to shape acute and chronic management of low back pain, but there is a dearth of information about the viewpoint of those suffering pain. This study attempted to bring the understanding of the back pain sufferer to the fore. Issues of quality of life, functional ability and the impact of back pain on their lifestyle were explored, along with the influence of contextual factors in relation to how back pain sufferers perceived themselves and how others perceived them. A narrative method was utilized to illuminate the journey with pain. Nine interviews were conducted, and the interpretation and presentation of the narratives generated was influenced by Ricoeur’s interpretative theory. Thematic analysis revealed that doctorability, agency, control, separation or acceptance of the pain and the concept of future life were key features within the narratives. The analysis highlighted that for the majority in this study pain arrived uninvited following a traumatic accident or incident, and back pain became a chronic condition. It was always unwanted and initially it was unexpected as the usual script for pain is one of a transient incapacity followed by recovery. It was precisely this deviation from the norm that resulted in difficulties for the people suffering the pain. Biographical differences did not appear to be identifiable in the themes discerned in the stories, nor in the overall structure.

Mutual Support is an in depth exploration of the role and impact of peer support by people with learning difficulties. Built on one of the seven aims of Centres for Independent Living, the project has constructed a model of peer support based on accounts of direct experiences from people with learning difficulties. The overall aim of the research was to construct and critique the Mutual Support model of peer support and people with learning difficulties. This thesis reflects the process of that construction. The overall aim was met through a research situation in which knowledge was constructed in the interaction between the researcher and participants. This provided an opportunity for people with learning difficulties to reflect upon their relationships with one another, and the emancipatory potential of that support. The focus of the research was two pre-existing settings involving people with learning difficulties supporting one another: a Theatre Company using Forum Drama to facilitate changes in attitudes and policy, and a course facilitated by people with learning difficulties who mentored small groups. Methods used within the research were based on an Inclusive Research process which prioritises meaningful research interaction that is accessible and guided by participants. The research process intertwined meetings with advisory groups, and contact with other local groups of people with learning difficulties, with formal data collection within the two main settings. One to one experienced-based narrative interviews with people from the two main settings provided multiple opportunities for participants to speak about their experiences of peer support. These interviews formed the data used in formal analysis, which was a continual process, with subsequent interviews being based on views previously expressed. A further comprehensive descriptive content analysis of data, using the tools of Nvivo8 and mind-mapping, took place prior to the outputs of the whole project being evaluated during group sessions with those who had taken part. The emerging model is one of collective support which challenges assumptions about the role and impact of people with learning difficulties supporting one another and their capacity to engage in insightful interpersonal interaction. Mutual Support has the potential to break down barriers to inclusion. Mutual Support also demonstrates the value that people with learning difficulties place on giving and receiving support from one another. The outputs of Mutual Support include contribution to current debate in the areas of service user involvement, inclusive research, and the academic field of Disability Studies.