No description available.

Previously cloned bacteriophage encoded HyIP 1 from the genome sequenced organism Streptococcus pyogenes SF370 was expressed in Escherichia coli and purified to homogeneity. The protein, previously assigned to glycoside hydrolase family 69 (GH69) was biochemically recharacterised as a polysaccharide lyase and reassigned to family 16 (PL16). The enzyme demonstrated a Km of 1.47 mg m1-1 and a kcat of 7.2 s-1. Biochemically the enzyme had an optimum pH of 6.5 and temperature of 37 °C. The enzyme required no additional divalent ions for catalysis. The enzyme demonstrated strict substrate specificity only degrading hyaluronate and with no activity against related substrates chondroitin 4 sulphate and chondroitin 6 sulphate. HPAEC indicated the HylP1 had an endo mechanism of cleavage producing a range of differently sized oligosaccharides with the smallest being a tetramer. Site directed mutagenesis revealed a role for residues D157 and Y169 with substitution of these residues with alanine resulted in a 88.5% and 91.9 % loss of activity respectively. The location of these residues within the solved structure of HylP1 falls within the triple stranded (3 helix formed by the trimerised protein. This region of the protein was cloned, expressed and characterised and demonstrated similar kinetics as the full length protein (Km of 0.53 mg m1-1 and keat of 11.1 s-1). The activity of the enzyme when compared to other hyaluronate lyases shows it to be relatively inefficient yet when compared to other bacteriophage encoded hyaluronate lyases, HylP 1 was very similar. The proposed role of these bacteriophage encoded hyaluronate lyases is one of degradation of the hyaluronate capsule surrounding the streptococcal cells to allow for penetration of the bacteriophage during infection. Using the sequence of HylP1 the recently completed genome of Streptococcus equi was searched using bioinformatics tool BLAST. This revealed the presence of a protein, SEQ2045, sharing 85 % identity with HylPl. The protein was cloned and expressed in E. colt and biochemically characterised as a hyaluronate lyase. The enzyme demonstrated a Km of 2.05 mg m1-1 and a kcat of 6.2 s-1 which when compared to those of Hy1P 1 is suggestive that the two enzymes are strongly related. The enzyme had an optimum pH of 6.5 and temperature of 37 °C and like Hy1P 1 demonstrated only activity against hyaluronate and had an endo mechanism of cleavage with the smallest product of digestion being a tetramer. Site directed mutagenesis of the same residues as in HylP1 again yielded reduced activity (91.3 % and 87.6 % respectively). Bioinformatic analysis of the genome of S. equi was performed by BLAST searching with the proposed gene sequences of S. equi flanking SEQ2045. This allowed for the production of a prophage map which shows distinct similarities to the prophage map of S. pyogenes suggesting both may be of the same origin. Purified SEQ2045 was used in western blot analysis with S. equi convalescent horse serum. A strong positive reaction demonstrated a possible role for SEQ2045 during an infection suggesting that the bacteria have acquired this enzyme as a potential virulence factor by horizontal gene transfer. This presents a useful opportunity for the study of both S. pyogenes and S. equi infection process and the role of bacteriophage by the use of S. equi as a model for S. pyogenes.

Anomalies noticed in midwifery practice prompted a review of literature around progress in labour after Loop Excision of the Transformation Zone (LETZ). The previous research was equivocal. This study investigates the pattern and progress of women's labours following LETZ, and the effect of the experience on women. It also explores the basis of clinical decisions made by midwives. It takes place in North East England and involves women giving birth in 2004-2005. The case study strategy brings together quantitative retrospective case control results, qualitative analysis of semi-structured interviews from women and midwives, and guidelines and protocols surrounding assessment and management of labour. The quantitative data is from 111 women after LETZ and 214 controls, reducing to 94 and 150 respectively after removal of cases with confounding variables. Interviews are from four women, three individual midwives caring for one of the women and a focus group of four midwives. In the interviews, women gave no thought to how their births may be affected by LETZ, as no one had discussed the possibility with them. Women having their second baby after LETZ take less time to reach full dilatation; a result related to grade of Cervical Intraepithelial Neoplasia. Women having their first baby after LETZ are 1.8 times more likely to be admitted later in their labours, an important emergent issue that needs further research. Women having their first or second baby after LETZ are 2.3 times more likely to have a premature birth, confirming indications in previous research. This study for the first time reveals important differences in pattern and progress of labour after LETZ. It adds to our knowledge of premature birth after LETZ. Midwives, obstetricians and colposcopists need to incorporate the study results into counselling of women before LETZ and during any pregnancy after LETZ.

Innovations in cancer care requiring intensive support, and improved cancer patient survival in and out of critical care, have led to greater numbers of cancer patients than ever accessing critical care. Of these, however, a fair proportion will die. Current research points to around one in six patients dying in general critical care units and even higher numbers for cancer patients. End-of-life care (EOLC) for critically ill patients is problematic and rarely addressed beyond satisfaction or chart review studies, while palliative care is an established domain in cancer. It is not known whether dying, critically ill cancer patients experience good EOLC. In the context of a cancer critical care unit, this thesis explores the provision of EOLC for cancer patients in a critical care unit. Exploring measures for comfort care and palliative principles of care helped identify what is important for patients and families, and what those measures meant for all participants. The diagnosis of cancer and how it impacts on EOLC provision for critically ill cancer patients was also explored from the perspective of patients, families, doctors and nurses. A Heideggerian phenomenological interview approach was undertaken, in order to gain personal experiences. Families of those patients who died after decisions to forgo life-sustaining treatment (DFLSTs) were interviewed. Patients who have experienced critical care were also interviewed, since patients‘ views about EOL care provision are very rarely explored. Doctors and nurses also contribute their vision for, and experiences of, EOL care in a cancer critical care unit. Thirty one interviews with 37 participants were carried out. Cancer prognosis together with critical illness prognosis contributed to difficulties in deciding to move to, and enact EOLC. The nursing voice in DFLSTs was minimal and their role in EOLC depended on experience and confidence. Achieving a good death was possible through caring activities that made best use of technology to prevent prolonged dying. EOLC was an emotive experience. Decision-making and EOLC could be difficult to separate out which, in turn, affects prospects for EOLC. A continuum of dying in cancer critical illness is presented with different participants‘ experiences along that continuum. Three main themes included: Dual Prognostication; The Meaning of Decision-Making; and Care Practices at EOL: Choreographing a Good Death with two organising themes: Thinking the Unthinkable and Involvement in Care. These themes outlined the essence of moving along a continuum toward patients‘ deaths and the impact that had on opportunities for care and a good death. Nurses could use the care of patients dying in critical care as an opportunity to develop specialist knowledge and lead in care, but this requires mastery and reconciliation of both technology and EOLC. This work builds on Seymour‘s (2001) theory of a negotiated and natural death related to achieving a good death in critical care. Trajectories of dying, part of Seymour‘s (2001) theory, are extrapolated on with reference to Glaser and Strauss (1965) and Lofland (1978)‘s theories on dying trajectories. Nursing theory is developed through examination of Falk Rafael‘s (1996) and Locsin‘s (1998) theories of empowered caring. Implications and propositions are presented for nursing and wider practice around EOL care for critically ill cancer patients.

Although parents’ involvement has been identified as important for interventions for their children and young people, little research is conducted to explore parental management strategies for anxiety in children and young people with mild to moderate learning disabilities (LD) and autism. Given that children and young people with LD and autism are more vulnerable to experience anxiety, the overall purpose of this thesis was to identify the prevalence rates of anxiety and explore management strategies for anxiety in children and young people with LD and autism. Three studies were conducted to identify prevalence of anxiety and develop and implement a parental programme of management strategies for anxiety in children and young people with LD and autism. In the first study of this research “Screening study”, 150 children and young people with LD and autism were screened for anxiety using Reiss Scale and Glasgow Anxiety Scale. Results from the screening indicated that children and young people with LD and autism have high prevalence rates of anxiety (32.6%). The second study of this research “Developing a programme” was to develop a programme of management strategies for anxiety from parents’, carer’s, teachers’ and health professionals’ views. In this study parents and teachers were interviewed regarding the management strategies they use to manage their children’s / young people’s anxiety. Results from 34 interviews revealed that sixteen different management strategies were identified by parents and teachers of children and young people with LD and autism. Following this, the management strategies identified by parents, carers and teachers were discussed with an expert panel of health professionals to develop a parental programme “Calm Child Programme” of the most useful and appropriate strategies for those children and young people. The Calm Child Programme was developed using Delphi method to identify a consensus approach with professionals. This consisted of ten useful management strategies for parents to manage their children’s and young people’s anxiety. Finally, the third study of this research “Implementing the calm child programme” was a pilot study of the implementation for the Calm Child Programme. This programme of parental management strategies for anxiety was piloted with seven parents of children and young people with LD and autism. The results from the pilot study indicate reduction in the children’s and young people’s anxiety after its use by parents. Parents reported that the developed strategies were useful in managing their children’s and young people’s anxiety. This research has two key contributions to knowledge. Firstly, this research highlighted the theme of anxiety of children and young people with LD and autism. The identification of high prevalence of anxiety in this population indicates the need for management strategies. Secondly, this research aimed to improve the paucity of autism specific research as it relates to anxiety management strategies in children and young people with autism. This research explored a new programme of parental management strategies for anxiety in children and young people with LD and autism. This research has contributed to our understanding of using systematic and evidence based management approaches in supporting and involving parents.

Complementary and Alternative Medicine (CAM) has long been used in the Far East to aid in the recovery and prevention of illness. Ginseng, an over-the-counter herbal product in the UK, is amongst these herbal CAMs currently available to the general public. Ginseng is renowned for its rejuvenating properties and its purported ability to aid cognitive function and well-being. Despite the huge global market for ginseng there is little in the way of human research, utilising standardised ginseng extracts and well controlled methodology to support many of these claims. Additionally, ginseng's underlying mechanisms of action are poorly understood. The present thesis documents 5 double-blind, placebo-controlled, cross-over trials investigating the effects of Panax ginseng, following acute and chronic ingestion, on behaviour, mood and indices of glucose regulation in young healthy volunteers. The results of the five studies making this thesis suggest that both acute and chronic dosing with Panax ginseng is capable of modulating mood and cognitive performance in healthy young volunteers. Chapters 2 and 3 also demonstrate, for the first time, Panax ginseng's ability to modulate blood glucose levels following a single acute dose in overnight fasted healthy volunteers. In chapters 2 and 3, significant reductions in blood glucose levels and concomitant improvements in mental arithmetic (working memory) performance were reported. Chapter 4 revealed for the first time Panax ginseng's positive effects on traditional measures of working memory, thus posing the suggestion that previous failures to report working memory effects (using traditional working memory tasks) may have been due to poor task selection. Chapter 5 revealed an unexpected superimposed relationship between chronic and acute ingestion of Panax ginseng. The pattern of results suggests that following chronic dosing, an acute dose can further modulate cognition and mood (suggestive of a psychological dependence). The final chapter documents a different profile of cognitive and mood effects following a non-standardised Panax ginseng extract, thus highlighting the need for caution when generalising results across ginseng types and beyond the specific parameters of the methodologies utilised in any given study. Methodological differences between studies may go some way in explaining the inconsistent data patterns reported between studies, research groups and ginseng extracts. These data further highlight the need for well-controlled studies utilising standardised ginseng extracts and the need for the integration of 'theory driven' research in order to fractionate any behavioural effect. Such methodologies will inevitably lead to greater consistency between behavioural studies, at least in the first instance within the restricted population of volunteers utilised in the present thesis.

Examination of the literature suggests that relationships between psychiatric staff and the families of service-users are often characterized as unsatisfactory, and that psychiatric hospitals provide an unconducive environment for the implementation of family and network-orientated approaches. This research focused upon the development in an adult psychiatric admissions unit of family- staff network meetings, that occurred when a person entered hospital. These network meetings, or 'reception meetings', were strongly influenced by the Finnish social network approach (Seikkula, Alakare & Aaltonen 2001), which focuses upon the creation of open dialogue between the service-user, family members and professionals. An overall aim of the study was to generate a body of practice-based narrative accounts which might act as a catalyst to practice developments in this field. Drawing upon a postmodern methodological framework, a range of methods were employed to engage with the multiple voices of service-users, family members and staff in the research site. Data sources included semi-structured interviews, participant observation, secondary data, and material from a research diary that was maintained as a reflexive tool in relation to the practitioner research process. A 'voice-centred relational method' (Brown and Gilligan 1992) provided a heuristic device for guiding data interpretation which facilitated multiple readings from different perspectives. A subsequent stage of data-analysis entailed developing more general connecting themes from across the data-set. A number of themes are developed in the research, which principally relate to the tensions associated with the introduction of a relationally orientated, reflective approach to practice within an individually-based medico-psychiatric organizational context. Practice approaches are discussed which appeared to facilitate an ethos of 'safe uncertainty' (Mason 1993) within network meetings, which is a necessary condition for dialogue. Connections are also drawn between themes generated in this study and the broader contexts of government policy, professional and organizational development, and practice-based research.

Multiple Sclerosis (MS), the most common progressive neurological disease in young adults can take a relapsing remitting (RR) course especially in the early stages. There is a gap in knowledge in the application of self-management and self-efficacy with progressive long-term conditions. This research explored the experience of individuals with RRMS with particular focus on their attitudes to self-management and development of self-efficacy. The research addresses the question about engagement with self-management and self-efficacy influencing the journey of people with RRMS and their formal and informal carers. The research draws on the experiences, perspectives and understanding of the social processes and reality through interaction. Using grounded theory for generation of the themes captured from people with RRMS, their partners/carers and professionals involved in their care. The research design around a conceptual framework, used longitudinal studies capturing the experiences of people with RRMS and their partners/carers, through individual interviews and self-efficacy qualitative questionnaires over eight monthly meetings. The research sample of people with RRMS was three male and three female with an average age of 44.5 years, a mean duration with RRMS for 9.6 years. Professionals shared their perceptions and experiences through a focus group and individual interviews. Personal and reflective diaries kept by the researcher of events throughout the study enabled decision and audit trials to contribute to the rigour of the research. The data analysis has generated a number of themes that have been developed and presented throughout the thesis. The research process has generated new theory around the knowledge of and experiences of the three groups of research participants. Fluctuations of living with RRMS across roller coaster journeys, with transitional processes of daily changes and meanings brought threats and challenges. These highlight the importance of self-efficacy and self-control, of coping with uncertainty and unpredictability, through engaging with self-management behaviours. These enhanced perceptions of self- determination, positiveness, independence, quality of life and well-being. The transformation of uncertainty secured hope, opportunities and embracing RRMS through coping mechanisms and self-confidence. Formal and informal support was required where the People with RRMS deemed appropriate. The epistemological perspective has explored power as a multilayered and dynamic concept with different knowledge bases and issues that need addressing prior to successful partnership working. The research is original in terms of the groupings, the areas reviewed for this RRMS group and the potential for ongoing work, policy and practice changes both locally and nationally. Implications for practice, policy and further research were derived from the conclusions. In practice more empowerment, advice and information are required for people with RRMS to live independently. Policies for skills and knowledge training in these areas are required for shared decision-making and partnership working. Further research is required into utilisation of Integrated Care Management of ways of supporting independence, self-control and personalised plans for people with long- term conditions and their informal carers.

No description available.

Background: The recent advent of non-invasive functional magnetic resonance image (fMRI) has helped us understand how visual information is processed in the visual system, and the functional organising principles of high-order visual areas beyond striate cortex. In particular, evidence has been reported for a constellation of high-order visual areas that are highly specialised for the visual processing of different object domains such as faces, bodies, and tools. A number of accounts of the underlying principle of functional specialisation in high-order visual cortex propose that visual properties and object domain drive the category selectivity of these areas. However, recent evidence has challenged such accounts, showing that non-visual object properties and connectivity constraints between specialised brain networks can, in part, account for the visual system’s functional organisation. Methodology: Here I will use fMRI to examine how areas along the visual ventral stream and dorsal action stream process visually presented hands and tools. These categories are visually dissimilar but share similar functions. By using different statistical analyses, such as univariate group and single-subject region of interest (ROI) analyses, multivariate multivoxel pattern analyses, and functional connectivity analyses, I will investigate the topics of category-selectivity and the principles underlying the organisation of high-order visual areas in left occipitotemporal and left parietal cortex. Principle Findings: In the first part of this thesis I report novel evidence that, similar to socially relevant faces and bodies, the human high-order visual areas in left occipitotemporal and left parietal cortex houses areas that are selective for the visual processing of human hands. In the second part of this thesis, I show that the visual representation of hands and tools in these areas show large anatomical overlap and high similarity in the response patterns to these categories. As hands and tools differ in visual appearance and object domain yet share action-related properties, the results demonstrate that these category-selective responses in the visual system reflect responses to non-visual action-related object properties common to hands and tools rather than to purely visual properties or object domain. This proposition is further supported by evidence of selective functional connectivity patterns between hand/tool occipitotemporal and parietal areas. Conclusions/Significance: Overall these results indicate that high-order visual cortex is functionally organised to process both visual properties and non-visual object dimensions (e.g., action-related properties). I propose that this correspondence between hand and tool representations in ventral ‘visual’ and parietal ‘action’ areas is constrained by the necessity to connect visual object information to functionally-specific downstream networks (e.g., frontoparietal action network) to facilitate hand-tool action-related processing.