The role of STX6 in sporadic Creutzfeldt-Jakob disease and related neurodegenerative diseases

Sporadic Creutzfeldt-Jakob disease (sCJD) is the most common human prion disease, in which the cellular prion protein (PrPC) appears to spontaneously misfold and aggregate into a disease-associated form leading to invariably fatal neurodegeneration. Although there is clear genetic control in sCJD, no loci except PRNP (encoding PrPC) have previously been consistently significant in genetic studies. Through international collaboration a two-stage genome wide association study (GWAS) was conducted for sCJD risk analysing a total of 5,208 cases and 511,585 matched control individuals from 11 countries. This thesis describes the first replicated association of two novel loci alongside PRNP, with variants in or near STX6 (encoding syntaxin-6) and GAL3ST1 (encoding cerebroside sulfotransferase), as conferring risk for sCJD at genome-wide significance in populations with European ancestry. With a focus hereafter on elucidating the casual gene and functional mechanisms at the STX6 locus, fine-mapping and integration of publically-available expression quantitative trait loci databases provides evidence for increased STX6 expression in the caudate and putamen as driving this association. RNA silencing in immortalised cell lines and CRISPR-mediated gene knockout in human iPS-derived cortical neurons and mice were used to model the biological mechanism of syntaxin-6 in prion propagation and cell biology with a particular focus on its role in the endolysosomal system. Knockout and hemizygous expression of Stx6 in C57BL/6 mice modestly extends survival time following inoculation with mouse-adapted scrapie prion strains RML and ME7, supporting the genetic association of STX6 expression in patients. Identification of non-PRNP loci associated with sCJD risk provides new insight into disease mechanisms and offers potential additional targets for therapeutic intervention. Furthermore the previous implication of STX6 in related neurodegenerative diseases such as the tauopathy progressive supranuclear palsy indicates a possible shared role of syntaxin-6 in the pathobiology of other “prion-like” proteins and disorders

Assessment of the autonomic nervous system through the study of cardiovascular autonomic reflexes and their association with inflammation in three clinical settings.

Heart Rate Variability describes the beat-to-beat variation in heart rate arising from activity of the sympathetic and parasympathetic branches of the Autonomic Nervous System (ANS). Reduced ANS tone measured by reduced heart rate variability (HRV) is a powerful predictor of adverse diagnosis in patients and is associated with increased mortality. Published research suggests that inflammation has a deleterious effect on Autonomic Nervous System tone. This study aimed to: establish if mild inflammatory conditions are associated with changes in autonomic tone as defined by heart rate variability studies in the following conditions: a. Influenza vaccination b. Reduction in oesophageal inflammation c. Reduction in weight The aim of the first study was to assess the link between inflammation resulting from the influenza vaccination and the associated changes on heart rate variability. 71 healthy volunteers opting to have a routine influenza vaccination were investigated for potential changes in cardiovascular autonomic tone associated with the temporary inflammatory effects of an Influenza vaccination. A number of temporal and frequency domain parameters of heart rate and breathing were assessed 2-5 days prior to vaccination and 1-4 days post vaccination. A sub-group of 15 volunteers who reported significant symptomatic reaction to the vaccination for at least 24 hours following vaccination displayed a statistically significant (p=<0.02) reduction in five of the six HRV parameters obtained during metronome-guided breathing. There was no evidence of significant reduction in autonomic tone following vaccination in the full sample of 71 volunteers. The aim of the second study was to establish whether inflammation resulting from erosive or non-erosive oesophagitis caused by gastro-oesophageal reflux disease had any association with changes in heart rate variability. 12 volunteers with non-erosive oesophageal reflux disease (NERD) and 8 with erosive oesophageal reflux disease (ERD) were investigated for HRV after initial diagnosis under gastroscopy. HRV assessment was repeated following 8 weeks of treatment with a proton-pump inhibitor (PPI). Initial reflux symptoms and response to PPI treatment were assessed using the GERD Impact Scale questionnaire. All participants had effective symptom response to treatment and there was no significant difference insymptoms score between NERD and ERD groups. There was a small but statistically significant increase in HRV detected following PPI treatment in the ERD group (p=0.05). The aim of the third study was to assess the link between obesity / pro-inflammatory adiposity, weight loss and the associated changes in heart rate variability. 38 clinically obese volunteers (BMI 30-39) with a family history of diabetes were reviewed for HRV prior to and following a lifestyle intervention designed to reduce body weight and BMI. Volunteers underwent repeated HRV studies after 4 months and 8 months of treatment. Volunteers on average achieved a weight loss of 11.5% (±6.0). There were statistically significant changes in HRV parameters in sub-group A (BMI ≥36) and correlation of biochemical measures with weight loss. These results further elucidate the effect of mild inflammatory triggers on autonomic tone as measured by HRV. These effects and their significance are discussed in detail in this document. The significance of the ‘cholinergic anti-inflammatory pathway’ is discussed with respect to the inflammatory conditions investigated. Suggestions for further work are proposed. In conclusion it is entirely possible to measure subtle changes in heart rate variability associated with mild inflammation and that on the evidence presented here these changes in heart rate variability are hypothesised to be reversible. My original contribution to knowledge is: 1. Changes in heart rate variability are associated with low grade inflammation resulting from the Influenza vaccination, erosive oesophagitis and increased adiposity. 2. Measurement of subtle changes in autonomic tone, associated with inflammatory challenges is possible and concurs with other published research. 3. The level of HRV attenuation does appear to be linked to those with a higher level of inflammation. In each study the most significant results came from subgroups of volunteers either demonstrating: a higher level of symptom severity, erosive oesophagitis or were in a subgroup of participants with the highest BMI / adipose tissue. 4. In the early stages of reduced heart rate variability we see that concurrent reduction in inflammation is associated with an increase in autonomic tone

Made in Wales: a study of the realities of work in a super independent television production company

The PhD study provides an assessment of TV production work in context, examining the impact of the consolidation of the independent television sector on the experience of work. The recent consolidation of the sector in the UK has led to the emergence of super independent television production companies, more commonly known as ‘super indies’. Super indies are organisations formed by multiple takeovers, grouping together independent television producers into larger companies. Whilst the rise of super indies is well-documented, the realities of work in super indies have yet to be explored. The thesis addresses this gap by assessing the impact of the super indie institutional structure on the experience of work in television production. Based on an in-depth case study of a Welsh super indie, Oaks Productions Ltd (Oaks), the research draws on 63 semi-structured interviews with both Oaks’ managers and employees to evaluate the experience of work in this context. In doing so, the research investigates firstly, how the nature of television production work is impacted by key features of the super indie structure in the Welsh context, and secondly, how creative work is experienced in this setting. Fundamental to the study is the identification of a creative worker typology, relating to the ability of creative workers to access and mobilise a variety of different resources to navigate creative work in this setting. Finally, the thesis explores the meaning of television work, highlighting the important role of individual creative work features, as well as social, relational and non-work factors in the experience of meaningful work. The thesis contributes to the TV production literature, illustrating the diversity of TV production experiences in a single organisational context. Moreover, it contributes to the creative labour literature by demonstrating the entrenched power hierarchies in creative work settings, and the importance of embedded resources in shaping the experience of television production work

Longitudinal association between change in the neighbourhood built environment and the wellbeing of local residents in deprived areas: an observational study.

BACKGROUND: Features of the urban neighbourhood influence the physical, social and mental wellbeing of residents and communities. We explored the longitudinal association between change to the neighbourhood built environment and the wellbeing of local residents in deprived areas of Glasgow, Scotland. METHODS: A cohort of residents (n = 365; mean age 50 years; 44% male; 4.1% of the 9000 mailed surveys at baseline) responded to a postal survey in 2005 and 2013. Wellbeing was assessed with the mental (MCS-8) and physical (PCS-8) components of the SF-8 scale. We developed software to aid identification of visible changes in satellite imagery occurring over time. We then used a Geographical Information System to calculate the percentage change in the built environment occurring within an 800 m buffer of each participant's home. RESULTS: The median change in the neighbourhood built environment was 3% (interquartile range 6%). In the whole sample, physical wellbeing declined by 1.5 units on average, and mental wellbeing increased by 0.9 units, over time. In multivariable linear regression analyses, participants living in neighbourhoods with a greater amount of change in the built environment (unit change = 1%) experienced significantly reduced physical (PCS-8: -0.13, 95% CI -0.26 to 0.00) and mental (MCS-8: -0.16, 95% CI -0.31 to - 0.02) wellbeing over time compared to those living in neighbourhoods with less change. For mental wellbeing, a significant interaction by baseline perception of financial strain indicated a larger reduction in those experiencing greater financial strain (MCS-8: -0.22, 95% CI -0.39 to - 0.06). However, this relationship was reversed in those experiencing lower financial strain, whereby living in neighbourhoods with a greater amount of change was associated with significantly improved mental wellbeing over time (MCS-8: 0.38, 95% CI 0.04 to 0.72). CONCLUSIONS: Overall, we found some evidence that living in neighbourhoods experiencing higher levels of physical change worsened wellbeing in local residents. However, we found a stronger negative relationship in those with lower financial security and a positive relationship in those with higher financial security. This is one of few studies exploring the longitudinal relationship between the environment and health

Lignocellulose-Degrading Microbial Communities in Landfill Sites Represent a Repository of Unexplored Biomass- Degrading Diversity Emma

The microbial conversion of lignocellulosic biomass for biofuel production represents a renewable alternative to fossil fuels. However, the discovery of new microbial enzymes with high activity is critical for improving biomass conversion processes. While attempts to identify superior lignocellulose-degrading enzymes have focused predominantly on the animal gut, biomass-degrading communities in landfill sites represent an unexplored resource of hydrolytic enzymes for biomass conversion. Here, to address the paucity of information on biomass-degrading microbial diversity beyond the gastrointestinal tract, cellulose (cotton) “baits” were incubated in landfill leachate microcosms to enrich the landfill cellulolytic microbial community for taxonomic and functional characterization. Metagenome and 16S rRNA gene amplicon sequencing demonstrated the dominance of Firmicutes, Bacteroidetes, Spirochaetes, and Fibrobacteres in the landfill cellulolytic community. Functional metagenome analysis revealed 8,371 carbohydrate active enzymes (CAZymes) belonging to 244 CAZyme families. In addition to observing biomass-degrading enzymes of anaerobic bacterial “cellulosome” systems of members of the Firmicutes, we report the first detection of the Fibrobacter cellulase system and the Bacteroidetes polysaccharide utilization locus (PUL) in landfill sites. These data provide evidence for the presence of multiple mechanisms of biomass degradation in the landfill microbiome and highlight the extraordinary functional diversity of landfill microorganisms as a rich source of biomass-degrading enzymes of potential biotechnological significance

Incidence of medically attended paediatric burns across the UK

Objective: Childhood burns represent a burden on health services, yet the full extent of the problem is difficult to quantify. We estimated the annual UK incidence from primary care (PC), emergency attendances (EA), hospital admissions (HA) and deaths.Methods: The population was children (0-15 years), across England, Wales, Scotland and Northern Ireland (NI), with medically attended burns 2013-2015. Routinely collected data sources included PC attendances from Clinical Practice Research Datalink 2013-2015), EAs from Paediatric Emergency Research in the United Kingdom and Ireland (PERUKI, 2014) and National Health Services Wales Informatics Services, HAs from Hospital Episode Statistics, National Services Scotland and Social Services and Public Safety (2014), and mortality from the Office for National Statistics, National Records of Scotland and NI Statistics and Research Agency 2013-2015. The population denominators were based on Office for National Statistics mid-year population estimates.Results: The annual PC burns attendance was 16.1/10 000 persons at risk (95% CI 15.6 to 16.6); EAs were 35.1/10 000 persons at risk (95% CI 34.7 to 35.5) in England and 28.9 (95% CI 27.5 to 30.3) in Wales. HAs ranged from 6.0/10 000 person at risk (95% CI 5.9 to 6.2) in England to 3.1 in Wales and Scotland (95% CI 2.7 to 3.8 and 2.7 to 3.5, respectively) and 2.8 (95% CI 2.4 to 3.4) in NI. In England, Wales and Scotland, 75% of HAs were aged < 5 years. Mortality was low with 0.1/1 000 000 persons at risk (95% CI 0.06 to 0.2).Conclusions: With an estimated 19 574 PC attendances, 37 703 EAs (England and Wales only), 6639 HAs and 1–6 childhood deaths annually, there is an urgent need to improve UK childhood burns prevention

Do OSCE assessments reflect clinical maturation of medical students? An evaluation of progression in core knowledge and competency domains

Introduction/Background Application of knowledge in clinical settings can be assessed using several different methods. However, constructing an effective evaluation system for an educational program is a challenging task. Purpose/Objectives In this study, we aimed to evaluate our medical curriculum by investigating medical students' progressive acquisition of core clinical competencies in their clinical years, using domain-based and behaviourally anchored checklist and global rating scales in the OSCE format. Issues/Questions for exploration/Methodology Three collaboratively developed OSCE cases were administered in the 2012 end of year OSCEs to all 3rd, 4th and 5th year medical students. These OSCEs assessed core clinical competencies Year of study effect on checklist and global scores as well as competency scores were analysed using the multivariate analysis of variance. Results Irrespective of year of study, the results showed similar patterns in student performance across the three OSCE stations. However, performance was significantly affected by year of study (p<0.01) and case difficulty (p<0.001). Discussion There was a progressive increase in performance, with the more senior students obtaining higher checklist and global scores. They also demonstrated better performance in more complex clinical skills such as diagnostic skills and investigation plan. Conclusions The use of domain-based and behaviourally anchored checklist and global rating scales enhance the validity of the OSCE in measuring progressive clinical competence. It also enables identification of problem areas and provides feedback to faculty on the efficacy of the curriculum