Objectives: The ability to regulate affect in the face of stress has implications for recovery and chronicity in complex mental health problems such as schizophrenia and borderline personality disorder. In addition to adaptive integrating and maladaptive sealing over recovery styles it may be possible to delineate a further maladaptive recovery style of “ruminative preoccupation”. In addition, the capacity to compassionately relate to self and others may be linked to an recovery trajectories. The current study presents data on the utility of a Narrative Compassion Scale for recovery in a mixed clinical sample of individuals with diagnoses of psychotic disorder (with or without interpersonal violence) and Borderline Personality Disorder Design: A cross-sectional mixed methods design was used with a within subjects condition and three between subjects groups Methods: Forty-Three individuals were interviewed and transcripts coded with the Narrative Compassion Scale (NCS). Self-report measures of compassion, attachment, interpersonal problems and symptoms were completed. Symptomatology was also measured. Results: Three recovery styles were identified. Compassion was strongly positively correlated with Integration; and negatively correlated with Sealing Over. NCS compassion was unrelated to self-reported compassion, symptoms, interpersonal problems or attachment. Differential patterns of recovery emerged between clinical groups, with lower preoccupation and higher sealing-over in the psychosis with history of interpersonal violence group. Conclusions: The NCS is a promising narrative measure of recovery and compassionate responding. Implications are discussed in terms of a transdiagnostic understanding of recovery processes.

The αVβ5 integrin is a member of integrin family that binds to different ligands such as vitronectin, fibronectin and soluble CD23 in order to mediate different biological responses such as cell growth, adhesion and metastasis. It is expressed by B cell precursors and by different acute lymphocytic leukaemia cell lines such as SMS-SB cells. This thesis is an attempt to explain how the sCD23- αVβ5 integrin interaction stimulates SMS-SB cell growth and to study the role of the αVβ5 integrin and other receptors such as PDGF receptor and CXCR4 in B cell development in the bone marrow. The maturation and differentiation of B-cells occur due to several factors that impact on gene expression in its development program. This program is divided into two main phases, the antigen-independent B-cell development phase and antigen-dependent B-cell development phase, respectively. The antigen - independent phase of B cell development starts from the pluripotent haemopoietic stem cell (PHSC) and progresses through several successive stages which are identified by somatic recombination and rearrangement of both heavy and light chain genes. Soluble CD23 and LP (a synthetic peptide derived from soluble CD23) significantly stimulate SMS-SB growth while a smaller growth stimulation is caused by either SDF1-α or PDGFAB. There are different signalling targets involved in the αVβ5 integrin-mediated proliferation due to its binding to either sCD23 or LP. These ligands enhance the association between the αVβ5 integrin and the PDGF receptor which promote the phosphorylation of both Jak2 and STAT5. Moreover, cell growth was reduced and the phosphorylation of Jak2 and STAT5 was also knocked down with using either PDGF receptor inhibitor (AG1295) or Jak2 inhibitor (AG490). Both soluble CD23 and LP activate the STAT5-DNA binding and strongly increase its transcriptional activity. In addition, both ligands induce the phosphorylation of other different substrates such as STAT2, c-Src, c-yes and AMPKα2 which might be related to cell growth stimulation. The αVβ5 integrin ligands also promote the phosphorylation of ERK1/2, p90RSK and activate a SRF transfected reporter gene. However, ERK1/2 and p90RSK phosphorylation was completely blocked by the specific MEK inhibitor (U0126). In similar context, SDF1-α stimulates the transcriptional activity of SRF but not STAT5 while PDGFAB does the opposite. Finally, soluble CD23 induces the proliferation of 697 and BAF03 which are other pre-B cell line models. These data suggest that the αVβ5 integrin-ligated ligands stimulate SMS-SB cell growth by promoting different signalling pathways, mainly Jak2/STAT5 and MEK/ERK1/2 pathway. Further work is required to determine the role of STAT5, p90RSK, c-Src and SRF in stimulating either the proliferation or apoptosis that promoted by the αVβ5 integrin-sCD23 interaction and to investigate the relationship between the activation of these targets.

The purpose of this thesis is to explore how differing circumstances came together to help or hinder the autonomous practice of midwives in Scotland between 1916 and 1983 when the Central Midwives Board (CMB) oversaw their training and practice. The thesis includes an examination of the records of the CMB for Scotland from 1916 to 1983 and, through oral testimonies, the work of practising midwives during the same period. The thesis is divided into two parts. Part I, comprising five chapters, explores the work of the CMB from 1916 to 1983. This includes: an examination of the issues surrounding the 1902 Midwives Act which did not apply to Scotland, and the campaign for, and opposition to, a similar Act for Scotland, passed in 1915; the constitution and early activities of the CMB which the 1915 Midwives (Scotland) Act established to oversee the enrolment, training and practice of midwives; a discussion of the national concern over the Maternal Mortality Rate in the 1920s and 1930s, subsequent Government reports and legislation and the CMB’s responses to these issues; an examination of the CMB’s work during the time of the World War II, the shortage of midwives, and the changes the National Health Service administration made to midwifery in Scotland. Finally, Part I examines the last decades of the CMB’s existence, including its response to changes in midwifery management, education, practice and statute. Part II, comprising three chapters, focuses on the practice of midwives in Scotland during the period through the aspects of antenatal, intranatal and postnatal care. Each chapter uses evidence from oral testimonies of midwives working within the framework established by relevant Acts and the CMB. Part II illuminates the contrast between the work of the CMB and the world of hands-on midwifery practice.

The aim of this project was to evaluate the use of LC-MS in the forensic toxicology setting. To investigate if its use could solve problems encountered while using other instrumentation including downtime, limits of detection, chromatography and specificity. Benzodiazepines as a class, are known for their undesirable chromatographic behaviour when using GC. Using LC-MS two models were developed for the analysis of these drugs in whole blood. Increasing awareness of drug facilitated sexual assault, has led to an increase in the number of cases to be analysed for drugs such as Rohypnol®. This prompted the development and validation of a sensitive and specific method for its detection in blood using solid phase extraction and liquid chromatography – mass spectrometry. To enhance the qualitative data and cope with degraded samples, the use of LC-MS was used in the analysis of diazepam and its three metabolites. A method was developed and validated and is now used in the routine analysis of blood samples in the laboratory. The LC-MS proved invaluable in the analysis of sildenafil, Viagra®, in a post-mortem blood sample received in the laboratory. A single quadrupole mass spectrometer was used to develop and validate a method to determine if the dose was therapeutic or toxic. The use of oral fluid as an alternative specimen to blood or urine was investigated. This was through the British Roadside Impairment Test Evaluation (BRITE) project. LC-MS in combination with GC-MS was used to screen for over 50 licit and illicit drugs in 1mL of oral fluid. LC-MS-MS was then used to identify and quantitated 21 drugs of abuse and their metabolites using a similar sample size. The use of one extraction and the reliability of LC-MS proved invaluable in these projects.

Introduction Adults with intellectual disabilities account for a minor proportion of the population, with reported prevalence rates in developed countries in the range of 3-6 per 1000 adults (Beange & Taplin, 1996; McGrother et al, 2002; McConkey et al, 2006) but have very high health needs and thus make up a proportionally larger section of the population with illness. Although it has been demonstrated that adults with intellectual disabilities have a higher prevalence of mental ill-health when compared to that reported for the general population (Bailey, 2008; Hassiotis et al, 2008; Cooper et al, 2007; Cooper & Bailey, 2001; Lund, 1985a; Corbett,1979), and some studies have shown a degree of persistence of behavioural problems and affective symptoms over time (Thompson & Reid, 2002; Collishaw et al, 2003), there is insufficient evidence to answer the question of whether this high prevalence is due to a high level of enduring mental ill-health or a high incidence of mental health, or indeed a combination of the two. To date, three studies (Holland et al, 2000; Zigman et al, 2004; van Schrojenstein Lantman-de Valk et al, 1997) have measured the incidence of dementia and only one study (van Schrojenstein Lantman-de Valk et al, 1997) has attempted to measure the incidence of affective disorder. No study has measured the overall incidence of mental ill-health in this population. Similarly, although the population based prevalence studies by Deb et al (2001a), Cooper et al (2007) and Bailey (2008) have identified some associations with mental ill-health (female gender, severe intellectual disabilities, past psychiatric history, not living with a family carer, smoking, life events, urinary incontinence and not having immobility), it is unknown whether some of these factors are cause or effect. No study to date has identified any adult risk factors for the onset of all types of mental-ill health in adults with all levels and causes of intellectual disabilities. As a consequence, our current knowledge of the epidemiology of mental ill-health in this population is limited and almost non-existent with regard to incidence and predictive factors of mental ill-health. The aim of this study was to measure the 2 year incidence rate of all types of mental ill-health in a population based sample of adults with intellectual disabilities with sufficient numbers to allow investigation of possible predictive factors for the onset of mental ill-health and examine the 2 year chronicity of mental-ill health in this population. Methods A large scale population based study of the prevalence of mental ill-health in adults with intellectual disabilities living in Glasgow, undertaken during 2002-2003 (Cooper et al, 2007), provided the opportunity to carry out a prospective longitudinal cohort design study, with the prevalence study providing the sample and baseline data. The sample size was 651 with a cohort retention rate of 70%. All participants were assessed using a two stage process (screening then detailed psychiatric assessment of potential cases) at baseline and at the 2 year follow-up interview. A modified version of the PAS-ADD checklist (Moss et al, 1998), with a reduced threshold for caseness to increase sensitivity, plus a problem behaviour checklist, pervasive developmental disorders checklist, and past 2 years mental health needs questionnaire was used to screen for mental ill-health occurring at any point during the two year follow up. All participants with identified episodes of mental ill-health were referred to the Glasgow University Centre for Excellence in Developmental Disabilities for detailed psychiatric assessment using PPS-LD (Cooper, 1997), checklists for problem behaviour, ADHD and pervasive developmental disorders and the Test for Severe Impairment (Albert & Cohen, 1992) (for possible dementia) and consensus diagnosis according to clinician, DC-LD, ICD-10-DCR and DSM-IV–TR criteria. Incidence and recovery rates were calculated. Standardised incident ratios were calculated by comparing the findings with reported rates for the general population. Stepwise binary logistic regression was used to examine factors independently related to the incidence and chronicity of mental ill-health. Key results The two year incidence of episodes of mental ill-health in adults with intellectual disabilities according to clinical diagnosis was 16.3%. This incidence rate is similar to the incidence rate of mental disorders in the general population but the type and proportion of individual disorders that accounted for this rate was different. Approximately 20% of this incidence rate was accounted for by problem behaviour, the incidence of psychosis, bipolar affective disorder and early onset dementia was very much higher than that reported for the general population with standardised incident ratios of 9.93 (95% CI 2.05-29.02), 100.20 (95% CI 12.14-361.96) and 66.67 (95% CI 18.16-170.69) respectively. The incidence of substance misuse and anxiety disorders was lower than that reported for the general population with standardised incident ratios of 0.04 (95% CI 0.00-0.24) and 0.17 (95% CI 0.06-0.37) respectively, although the lowered rate of anxiety disorders might be due to the methodological limitations of this study. Factors found to be predictive of episodes of mental ill-health (excluding problem behaviour, dementia, and delirium) were, in order of decreasing strength of association: not living with a family carer, not having immobility, mental ill-health in the past, more severe intellectual disabilities, abuse/adversity in adulthood, and urinary incontinence. A high level of chronic mental ill-health was found with a 2 year recovery rate of only 32.5%. Factors identified as associated with the endurance of mental ill-health (excluding problem behaviours) in adults with intellectual disabilities were, in decreasing order of strength of association: problem behaviour, not having Down’s syndrome, not having immobility and smoking. Conclusions The overall incidence of mental ill-health in adults with intellectual disabilities is similar to that reported for the general population but the type and proportion of disorders accounting for this is different. There is high level of enduring mental ill-health in adults with intellectual disabilities. It appears that the high point prevalence of mental ill-health in adults with intellectual disabilities compared to the general population is accounted for more by a higher level of endurance of mental ill-health than by a higher incidence. The identification of risk factors for the onset of mental ill-health means that hypothesis based studies, leading on to the development of interventions and then randomised controlled trials are now possible.

Human T cells expressing CD56 are capable of tumour cell lysis following activation with interleukin-2 but their role in viral immunity has been less well studied. The work described in this thesis aimed to investigate CD56+ T-cells in relation to cytomegalovirus infection in healthy subjects and kidney transplant patients (KTPs). Proportions of CD56+ T cells were found to be highly significantly increased in healthy cytomegalovirus-seropositive (CMV+) compared to cytomegalovirus-seronegative (CMV-) subjects (8.38% ± 0.33 versus 3.29%± 0.33; P < 0.0001). In donor CMV-/recipient CMV- (D-/R-)- KTPs levels of CD56+ T cells were 1.9% ±0.35 versus 5.42% ±1.01 in D+/R- patients and 5.11% ±0.69 in R+ patients (P 0.0247 and < 0.0001 respectively). CD56+ T cells in both healthy CMV+ subjects and KTPs expressed markers of effector memory-RA T-cells (TEMRA) while in healthy CMV- subjects and D-/R- KTPs the phenotype was predominantly that of naïve T-cells. Other surface markers, CD8, CD4, CD58, CD57, CD94 and NKG2C were expressed by a significantly higher proportion of CD56+ T-cells in healthy CMV+ than CMV- subjects. Functional studies showed levels of pro-inflammatory cytokines IFN-γ and TNF-α, as well as granzyme B and CD107a were significantly higher in CD56+ T-cells from CMV+ than CMV- subjects following stimulation with CMV antigens. This also resulted in higher levels of proliferation in CD56+ T-cells from CMV+ than CMV- subjects. Using class I HLA pentamers, it was found that CD56+ T-cells from CMV+ subjects contained similar proportions of antigen-specific CD8+ T cells to CD56- T cells in healthy donors of several different HLA types. A comprehensive gene expression profile by microarrays of CMV-stimulated sorted CD56+ T-cells was conducted which showed that expression of 106 genes involved in innate and adaptive immune responses was significantly changed, almost all being increased. Some of these changes were validated via RT-PCR and flow cytometry, the latter indicating higher ISG-15 protein expression in response to CMV stimulation in CMV+ than CMV- subjects. Overall, these differences may reflect the expansion and enhanced functional activity of CMV-specific CD56+ memory T cells. In view of the link between CD56 expression and T-cell cytotoxic function, this strongly implicates CD56+ T cells as being an important component of the cytotoxic T-cell response to CMV.

Malaria control relies heavily on the use of insecticides, especially the pyrethroids, for control interventions such as Long Lasting Insecticide Nets (LLINs) and Indoor Residual Spraying (ITNs). However, widespread resistance to insecticides in major malaria vectors, such as An. funestus is threatening to derail these control tools. To design and implement suitable resistance management strategies which will ensure the continued effectiveness of these control tools it is necessary to elucidate the molecular basis of the resistance. In An. funestus, resistance is mainly metabolic with the duplicated P450s CYP6P9a and CYP6P9b implicated as the major pyrethroid resistance genes. Despite the detection of these key resistance genes the detailed molecular mechanisms through which they confer pyrethroid resistance remain uncharacterised. Because CYP6P9a and CYP6P9b were shown to exhibit significant allelic variation between resistant and susceptible mosquitoes, we hypothesised that this allelic variation is potentially a key mechanism conferring pyrethroid resistance. Here, I characterised the role of these genes in the resistance to pyrethroids and identified other candidate genes which confer cross-resistance to non-pyrethroid insecticides. The role of allelic variation in pyrethroid resistance was investigated using polymorphism survery and in silico prediction of activity. Metabolic activities and efficiencies of allelic variants of CYP6P9a and CYP6P9b were investigated using fluorescent probes, metabolism assays and transgenic expression in D. melanogaster system. Pyrethroid resistance causative mutations were detected using the site-directed mutagenesis. Other candidate P450s that confer cross-resistance to pyrethroids carbamates and organochlorines were identified and characterised. This study revealed that CYP6P9a and CYP6P9b from resistant populations of An. funestus are undergoing directional selection with reduced genetic diversity and beneficial mutations selected, compared to the alleles from susceptible strain (FANG), which exhibited high genetic variation. Modelling and docking simulations predicted the alleles of CYP6P9a and CYP6P9b from the resistant strains all across Africa to metabolise pyrethroids with high efficiency while the susceptible alleles FANGCYP6P9a and FANGCYP6P9b were predicted to have low activity toward pyrethroids. Validation of the docking predictions with probes and metabolism assays established that the resistant alleles of CYP6P9a and CYP6P9b possess high activities toward pyrethroids with kinetic profiles significantly different (high affinity and catalytic efficiency) from those obtained from the FANG, indicating that allelic variation is playing a major role in pyrethroid resistance. These findings were further strengthened by results from transgenic expression with GAL4/UAS technology showing that flies expressing the resistant alleles of both genes were significantly more resistant to pyrethroids than those expressing the susceptible alleles. Using mutagenesis, three key residues (Val109, Asp335 and Asn384) from the resistant allele of CYP6P9b were established as the important amino acid changes responsible for resistance with impact on substrate channelling, possible enhancement of interaction with redox partners and inter-molecular hydrogen bonding interactions, respectively conferring high metabolic efficiency. The finding of these resistance markers make it possible to design a diagnostic tool that can allow detection and tracking of the resistant alleles in the field population of An. funestus across Africa. Other up-regulated P450s in multiple resistant populations from southern Africa were also characterised, revealing that pyrethroid resistance is mediated by other P450s as well: CYP6M7, CYP6Z1, CYP9J11 and CYP6AA4 all of which metabolise pyrethroids. CYP6Z1 and CYP9J11 are cross-resistance genes which metabolise bendiocarb, while CYP6Z1 metabolise DDT in addition. In conclusion, allelic variation is a key mechanism conferring pyrethroid resistance in An. funestus s.s. from sub-Saharan Africa. Key amino acid changes control pyrethroid resistance factors and these molecular markers can be used to design DNA-based diagnostic tests which will allow tracking of the resistance alleles in the field. Pyrethroid resistance is multi-genic in the field populations of An. funestus with other P450s involved apart from CYP6P9a and CYP6P9b. The finding of cross-resistance P450s is of concern to resistance management and should be taken into account when designing resistance management strategies.

The effect of stimulation with TSLP and KRN7000 on iNKT cell proliferation and cytokine production (S Salman) iNKT cells are small subset of T lymphocytes that bridge the innate and adaptive immunity. They have the ability to secrete large amount of cytokines upon stimulation with their glycolipid ligand such as αGalactosylceramide or KRN7000 and therefor skew the immune response towards Th1, Th2 or Th17. When in an inflammatory or infectious environment they also are target to signals from other cells that are involved in the immune response. Thymic stromal lymphopoietin (TSLP) is a cytokine that is produced by epithelial cells in allergic diseases and it promotes Th2 response. The hypothesis proposed here is that TSLP may affect iNKT cells directly on TSLPr that may be expressed by these cells and induce a Th2 type cytokines such as IL-4 and IL-13. Also the notion that the addition of TSLP to iNKT cells culture with KRN7000 may stimulate these cells and result in increased proliferation and cytokine production. The first approach to investigate the hypothesis was to optimise the experiment parameters such as the best glycolipid ligand with the optimal concentration and the optimal time point. After that the expression of some markers was investigated especially TSLPr both at day 0 and after 14 days stimulation. Finally the effect of adding TSLP to KRN7000 on iNKT cells (proliferation and cytokine production) in PBMC mixed culture and on enriched iNKT cells and iNKT cell lines were investigated. The results of this work demonstrated that iNKT cells express TSLPr, which is up regulated after stimulation with KRN7000 for 14 days. Furthermore, the addition of TSLP to KRN7000 in iNKT cells in PBMC cultures significantly increases their proliferation in the two healthy donors groups, high and low responders. It also increases IFN-γ production in both groups, IL-13 and IL-17 production in high responders. The cytokine production source was further confirmed to be from iNKT cells after enriching these cells and after using iNKT cells lines. In summary, TSLP and KRN7000 stimulate healthy human iNKT cells and significantly increase their proliferation and cytokine production.

Malaria remains one of the most debilitating tropical diseases with more than 90% of cases in Africa among children under five and pregnant women. Resistance observed against the main insecticides used in public health sector in major vectors such as Anopheles funestus is threatening the success of vector control interventions. To improve the design of suitable resistance management strategies, it is crucial to elucidate the underlining molecular basis of resistance or cross-resistance between insecticides and also establish patterns of gene flow between populations to predict the speed and direction of spread of resistance genes. To address these questions, this study has investigated the molecular basis of resistance to carbamates and cross/resistance to carbamates/pyrethroids in a population of An. funestus from Malawi. This study has revealed that metabolic resistance is the main mechanism driving carbamate resistance through the over-expression of Cytochrome P450 genes. Genome-wide microarray-based transcription analyses consistently revealed that the duplicated P450 genes CYP6P9a and CYP6P9b were among the most up-regulated genes ( > 2-fold; P < 0.01) in bendiocarb and pyrethroid resistant mosquitoes from Malawi. Other P450 genes were also associated with both carbamates resistance and a carbamates/pyrethroid cross-resistance, including CYP6Z1, CYP9J11. Analysis of polymorphism patterns of CYP6P9a and CYP6P9b genes between resistant and susceptible mosquitoes revealed a correlation with permethrin resistance but not for bendiocarb resistance. In addition a reduced penetration mechanism was shown to be likely involved in carbamate resistance through the elevated expression of several cuticular protein genes. An Africa-wide genetic structure of An. funestus populations was established using microsatellite markers. This study revealed that An. funestus populations from Malawi and Mozambique from southern Africa are genetically closer to each other than the rest of the other African populations from West, Central and East Africa. Two loci AFUB6 and FUNR that are located around same position as the CYP6P9a and CYP6P9b genes on the rp1 QTL on the 2R chromosome are under selection possibly because of pyrethroid resistance due to their low allele number and reduced heterozygosity. Statistical analysis revealed that both loci were under possible selection and sequencing analysis also revealed that CYP6P9a gene is under selection for the resistant An. funestus from southern Africa compared to more susceptible population from Cameroon. The possible cross-resistance observed in this study highlights the need to further elucidate resistance mechanisms before implementing suitable resistance management strategies such as rotation of insecticides to ensure a continued effectiveness of control interventions.

Type 2 Diabetes (T2D) affects over 246 million people worldwide. South Asians are the largest group affected and have the fastest growing prevalence. South Asians have been identified as at risk due to biomedical dispositions including central adiposity and insulin resistance. Lifestyle factors are also associated with T2D. However, the majority of research area tends to homogenise south Asians, consequently overlooking some of the subtle cultural differences between sub groups. As a result, British Indian Punjabi men have been neglected from research, despite being one of the largest groups at risk. This qualitative study examined the social and cultural factors which influence Indian Punjabi men’s health beliefs and risk perceptions of T2D. Twenty-four interviews were carried out with Indian Punjabi men from first and second generations. The data was analysed using a thematic analysis approach. The findings revealed the cultural significance associated with hospitality and drinking, which emerged as important socio-cultural practices. The findings also demonstrated generational differences in how the men perceived their risk of T2D in either individual or collective terms. The findings of this research study have the potential to make an important contribution to T2D care by highlighting changes to future care strategies.