The chaperonin proteins form a ubiquitous family of molecular chaperones and are absolutely required for correct folding or assembly of a subset of proteins in the cell. They are divided into two groups based on phylogeny: Group 1 found in bacteria and eukaryotic-organelles and Group 2 found in eukaryotic-cytoplasma and archaea. The two groups share a significant degree of conservation but differ in structure and mechanism and are believed to have evolved to serve specific client proteins. Using archaeal chaperonin from M. maripaludis (MmCCT) as a representative, we report here for the first time that a Group 2 chaperonin can partially replace the function of a Group 1 chaperonin from E. coli (GroEL). We have also identified and characterized two functional variants of MmCCT that show better GroEL complementation and have utilized them for a preliminary mutational analysis of potential client binding residues of MmCCT. We further demonstrate an initiative using a tagging approach for identification of bacterial proteins that interact with MmCCT in vivo. We suggest that our findings provide a novel platform for genetic dissection of MmCCT using a comparatively simple host, E. coli, which in turn can help identify properties of this archaeal chaperonin and provide insights for structure-function co-relations of Group 2 chaperonins in general.

Spiritual matters lie at the heart of a good dying. However, the shape and focus of spiritual care in palliative care has, I believe, shifted away from the original vision of hospice pioneers, such as Cicely Saunders. Assessing the value of spiritual care has become a priority to those who value a 'scientific' evidence-based practice. The issue this thesis addresses is whether there is a better 'artistic' way to evaluate the care given to dying people. This thesis describes the current landscape of care and then argues for spiritual artistry, as a way to reclaim 'inner space' and 'deep silences' as sites of sacred encounters. This radical move takes the focus of spiritual care away from both evaluative endeavours and also beyond the alternative meaning-making and narrative models, which are currently offered. I hope to shift the focus towards a relational spirituality in which greater attention is given to spiritual awakenings. Dying and grieving can both be viewed as the space of spiritual quests. The challenge is to discover a new way of seeing these complex and sometimes chaotic spiritual contexts in which important things occur which sometimes are beyond measurement and communication. An innovative methodology, spiritual autoethnography, which integrates creative arts, autoethnography and theology, is chosen to pursue this research. In the process of interrogating the silences enounterered in spiritual care at the end of life new insights and understandings are generated. I illustrate the deep silences that occur in times of trauma, shame, cognitive impairment, betrayal and grief, and how understanding these sheds light on marginalised areas rarely the focus of current models of spiritual care. However, the radically new insights gleaned from this research come from the construction of inner space and deep silences as a locus of the sacred. These heuristic constructs, offer a new framework to shape the role of hospice chaplain, and the delivery of spiritual care. In conclusion, spiritual artistry, founded on a poetics, is presented as enabling chaplains to inhabit 'inner space' and 'deep silences'; to say the unsayable and delight in the gifts that accompany griefs. This new understanding of the role of the hospice chaplain, and of the delivery of spiritual care, benefits all those confronting their mortality and their grief.

This study aimed to investigate any difference between the attributions parents made about their child’s sleep problem, in parents of children with an Autism Spectrum Disorder and parents of children with Down Syndrome. Seventy-six parents of children with an Autism Spectrum Disorder and fifty-two parents of children with Down Syndrome completed a series of questionnaires on-line, regarding their child’s sleep problem, their beliefs about their child’s sleep problem and the parent’s level of anxiety and depression. A significant difference was found between the groups on four of the causal items; other health problem, child’s emotional state, child’s personality and diet. Parents of children with Down Syndrome showed a higher level of agreement that their child’s sleep problem could be attributed to another health problem compared to parents of children with an Autism Spectrum Disorder. Parents of children with an Autism Spectrum Disorder showed a higher level of agreement that their child’s sleep problem could be attributed to their child’s personality, their child’s emotional state and their child’s diet compared to parents of children with DS. There was a high level of agreement across all parents that their child’s disability was a causal factor to their sleep problem and differences in attributions may reflect characteristics of the child’s diagnosis. The results are consistent with previous findings that parents view disability as an important causal factor to their child’s sleep problem and suggest a possible overlap in parent’s views of their child’s sleep problem and views of their disability.

Objectives: This study explores the relationships between challenging behaviour, emotional recognition, alexithymia and cognitive emotional regulation strategies in a population of people with learning disabilities. The Emotional Recognition Questionnaire was developed to measure an individual’s ability to identify the emotions they would feel in a given situation. One objective of this study was to assess the validity and reliability of the tool. Methodology: Cross-sectional data was collected from 96 participants with a learning disability and 95 of their carers. The service user participants completed the Emotional Recognition Questionnaire (ERQ), and adapted versions of the Alexithymia Questionnaire for Children (AQC) and the Cognitive Emotion Regulation Questionnaire for Children (CERQ-k). Carer participants completed the Checklist for Challenging Behaviour (CBC) and the Observer Alexithymia Scale (OAS). Correlational analyses were computed to identify relationships between the variables and linear regression was used to identify the predictive value of variables in relation to the main outcome variables of challenging behaviour frequency, management difficulty and severity. Finally, a between group analysis was conducted to compare the emotional recognition abilities of people with high frequency challenging behaviour with those with low or no challenging behaviour. Analyses were conducted to test the hypotheses. Results: No relationship was found between the ERQ and the AQC so the construct validity of the ERQ was not supported. The results highlighted significant negative associations between emotional recognition abilities and challenging behaviour frequency and management difficulty. Significant differences in emotional recognition abilities were found between people with high frequency challenging behaviour and those with low or no challenging behaviours. Observer rated alexithymia was significantly related to challenging behaviour frequency, management difficulty and severity. Cognitive emotional regulation strategies and service user measured alexithymia were not, however, related to challenging behaviour. Other relationships were found between service user rated alexithymia and the cognitive emotional regulation strategies of Catastrophizing and acceptance, and emotional recognition was negatively related to self-blame. Conclusions: Overall, the study suggests that emotional recognition and observer related alexithymia are important in understanding challenging behaviour presented by people with a learning disability. This has implications for clinical practice and further research. Additional research needs to be conducted to evaluate the construct validity and test-retest reliability of the Emotional Recognition Questionnaire.

Diabetes Mellitus (DM) has become one of the most important metabolic diseases that reduces one’s quality of life and doubles the risk of early death. Amongst the major complications linked to DM, diabetic retinopathy (DR) leads to gradual loss of vision and blindness. DR is now the second cause of certifiable blindness among the working age adults in the UK. The lifetime costs to the UK government are calculated to be up to £327,000 per person, with almost 50% of these costs being attributed to loss of productivity caused by visual impairment and blindness. The UK is one of the leading countries in the implementation of DR screening programmes. The latter rely heavily on fundus imaging and grading using trained experts and subsequent referral to hospital for further clinical examination and evaluation depending on the grade of retinopathy. It is now known that subtle, structural changes in the retina that are linked to diabetes can precede detectable vascular changes. The former can affect one’s colour vision and this offers the potential of using changes in chromatic sensitivity as an early biomarker of retinal disease. The first part of this thesis focuses on measuring chromatic sensitivity using the colour assessment and diagnosis (CAD) test in diabetic subjects with varying degrees of retinopathy. The severity of colour vision loss is graded in comparison to other factors that are normally linked to diabetes, such as the type, grade, control methods and duration. The results of this study reveal losses of both red/green and yellow/blue chromatic sensitivity in patients with diabetes, but the correlation with factors, normally associated with high risk of diabetes is low. The results from this study do, however, show that the magnitude of chromatic sensitivity losses correlates with the severity of diabetic retinopathy. The second, related study examines the effectiveness of intravitreal injection of a dexamethasone implant (Ozurdex) in patients with diabetic macular oedema (DMO) in stabilising and reducing loss of visual function and in particular the reduction in chromatic sensitivity up to 24 weeks. This treatment demonstrated efficacy in the treatment of chronic DMO and DMO which is resistant to anti-VEGF treatment. The results show that intravitreal treatment with Ozurdex causes improvement is visual acuity, central retinal thickness and significant improvement in red/green chromatic sensitivity.

Spinal cord injury (SCI) has a dramatic effect on the function and appearance of the body. The first chapter of this thesis is a literature review which focuses on studies with participants who have incurred an alteration to their appearance due to a health condition. The implications of these studies for the likely impact of Body Image (BI) in men with SCI are considered. Cognitive and disfigurement models of BI are reviewed and research evidence evaluated. Results reveal that whilst the extent of appearance change is an important predictor of BI dissatisfaction, further variation is explained by the strategies individuals use to cope with the alteration in their appearance. BI dissatisfaction appears to impact upon the psychosocial wellbeing of both men and women and can also affect physical health. Further research is therefore needed in order to explore the links between appearance change, methods of coping and BI dissatisfaction, particularly in men with SCI. The second chapter reports on a cross sectional study of 102 male participants with SCI, which revealed group differences in BI dissatisfaction between inpatients and outpatients, who are established in the community. BI investment was found to moderate the effect of BI evaluation on low mood. The use of maladaptive coping strategies was also found to moderate the relationship between BI dissatisfaction and BI quality of life. Evidence suggests that BI should be assessed and BI dissatisfaction monitored after discharge from rehabilitation hospitals. Clinical intervention could focus upon reducing the use of maladaptive coping strategies, in order to reduce the impact of BI dissatisfaction upon quality of life.

No description available.

Background: Pressure ulcers (PrUs) have a significant impact on health system expenditure and patient’s quality of life. It is a global problem. Many studies were undertaken in regard to PrU prevention and management. In Oman, no studies have been conducted to investigate nurses’ knowledge on prevention and management of PrUs. The purpose of this descriptive sequential explanatory mixed-method study was to explore the nurses’ level of knowledge in relation to prevention and management of PrUs in Oman. Methods: A mixed method design was used and the study was conducted over two Phases. In Phase I, a questionnaire was developed to explore nurses’ knowledge on PrU, policy, and resources. The main section of the questionnaire was the Pieper-Zulkowski Pressure Ulcer knowledge test (PZ-PUKT) which tests the knowledge on PrU. Another two sections were developed including questions about wound policy and resources available for PrU prevention and management in Oman. The questionnaire was distributed to nurses who were working in surgical, medical, orthopaedic, CCU, and ICU wards/units in seven hospitals. In Phase II study, semi-structured qualitative interviews were conducted with 16 of the questionnaire respondents. Interviews took approximately 30 minutes, were recorded and transcribed verbatim. Qualitative data were analysed using the Knowledge, Attitudes and Practice (KAP) model as the a priori framework. Results: In Phase I, 478 questionnaires were analysed. The knowledge test results showed the overall mean percent score for correctly answered questions was 51% suggesting a low level of knowledge. There was a significant relationship between nurses’ knowledge and age (P=0.001) and between knowledge and years of experience (P=0.001) with knowledge increasing with age and years of experience. In Phase II, four themes were identified from the interviews: knowledge, attitude, and practice (framework themes) and perception of role. Findings indicated positive and negative attitudes towards the care of PrUs. Some nurses stated feeling rewarded when they see wounds improving while others said they could not work with patients independently because they lacked the knowledge and the skills needed. There was variation in the management of PrU between hospitals. Both studies indicated that the wound management policy did not include enough information to guide nurses. Conclusion: Overall the nurses’ level of knowledge on PrU was relatively low. Most nurses were not familiar with wound management policy or different PrU prevention and management strategies. Nurses are aware of the risk of PrUs and try their best to manage them with the available resources however more training is required.

Transplantation of genetically engineered neural stem cells (NSCs) into sites of central nervous system (CNS) disease/injury is a promising strategy to promote repair of damaged tissue. However, translating this strategy into the clinic requires several challenges to be overcome including facilitating ‘combinatorial therapy’ (achieving multiple therapeutic goals – essential in CNS injury/disease). Nanotechnologies are emerging as multifunctional platforms capable of meeting this requirement. For example, magnetic particles (MPs) and implantable hydrogels offer several biomedical advantages for transplant populations, including: safe genetic manipulation; non-invasive cell tracking, via MRI; and safe and efficient accumulation of cells at sites of injury. However, the use of these nanotechnologies remains to be explored in detail for NSC transplantation therapies. In this thesis, it is shown that MPs can mediate gene delivery to NSCs grown as neurospheres and monolayers with the most efficient transfection efficiencies achieved using oscillating magnetofection protocols (9.4% and 32.2% respectively). In both culture systems, developed protocols had no effect on key regenerative properties of NSCs such as cell viability, proliferation, stemness and differentiation. Further, ‘magnetofected’ monolayer NSCs were shown to have survived and differentiated in a cerebellum slice model acting as host tissue, indicating safety of the procedures. It was also shown that assessing procedural safety and extent of transfection of magnetofection protocols may be feasible by employing mass spectrometry and proteomics analysis. It was also found that tailored enhancement of particle magnetite content offers a means to efficiently label NSCs, up to a maximum of 95.8%. Labelling procedures had no effect on cell viability, proliferation, stemness or differentiation. In addition, labelled cells could survive and differentiate in a slice model of spinal cord injury indicating safety of the labelling procedures. Functional labelling was also demonstrated by magnetic capture of labelled cells in an in vitro flow system. Hydrogels offer major advantages for delivery of transplant populations into injury sites. Here it was shown that an intraconstruct genetic engineering approach was feasible for NSCs cultured with a clinically translatable, collagen hydrogel system. Magnetofection protocols safely increased MP mediated transfection of NSCs grown in ‘2-D’ and ‘3-D’ hydrogel cultures.

More than 90 serotypes of Streptococcus pneumoniae are responsible for causing a number of invasive and non-invasive diseases in humans. To combat this pathogen there are two types of vaccines available namely pneumococcal polysaccharide vaccine (PPV) and pneumococcal conjugate vaccine (PCV). Although these vaccines have reduced the incidence and prevalence of pneumococcal diseases yet these vaccines are unable to curb the disease and have some potential drawbacks. The problems associated with PPV is inability to mount appropriate immune response in young children (<2 years) of age and with PCV, disadvantages are serotype replacement and affordability. These disadvantages have shifted the focus of attention on developing vaccines that can provide serotype independent protection and is cost effective and can be used in developing and under-developing countries as well. Pneumolysin (PLY) is one of the members of cholesterol dependent cytolysin (CDC) family and is one of the key virulence factors of the bacterium. PLY has a wide variety of functions but the two most important functions are the ability to lyse the host cell by forming pores and activating the complement pathway. Studies have shown recombinant PLY to be a good vaccine candidate; however PLY is toxic and cannot be used as a vaccine. A number of toxoid versions have been made targeting residues responsible for toxin’s lytic activity and tested in-vivo models. ∆6 PLY is a toxoid made by deleting two amino acid residues and seems to be a potential candidate to be included in next generation pneumococcal vaccines. ∆6 PLY is immunogenic and act as an adjuvant as well, however ∆6 PLY causes the aggregation of red blood cells and therefore cannot be used in the vaccine. The present study is carried out to identify the residue/s responsible for causing this agglutination behaviour. A number of molecular biology techniques were used in this study for making mutants in different backgrounds and were tested for their lytic and agglutination activity. The study identified the single residue in domain 4 causing aggregation of the red blood cells. The agglutination negative mutant was then tested at different concentrations through a series of experiments including hemagglutination assays, fluorescence microscopy and fluorescence activated cell sorter (FACS). The study also highlighted some important regions of the toxins responsible for maintaining structure of the toxin. This new toxoid seems to have the potential to be used in future pneumococcal vaccines alone, in combination with other pneumococcal proteins and/or polysaccharides.