Modelling FKRP and fukutin deficiency in zebrafish

PhD ThesisDeficiency in fukutin-related protein (FKRP) or fukutin results in aberrant glycosylation of -dystroglycan, a key receptor for basement membrane proteins. There is a broad spectrum of disorders associated with FKRP and fukutin deficiency, ranging from limb-girdle muscular dystrophy to congenital disorders such as muscle eye brain disease and Walker-Warburg syndrome (WWS). fkrp and fukutin were knocked down in the zebrafish with antisense morpholino oligonucleotides (MO). The fkrp, fukutin and dystroglycan MOs each produced a spectrum of comparable phenotypes. With each MO producing a comparable morphant phenotype on morphological examination, it was hypothesised that inferences could be made about similarities and differences in the fkrp, fukutin dystroglycan axis during zebrafish development. The morphants had abnormal muscle fibres, including disruptions of the vertical myosepta and sarcolemma. Disorganised retinal layering in the eyes was found in both fukutin and fkrp morphants. Dysplasia of the lens was observed in most fukutin morphants and some of the fkrp morphants with a severe phenotype. Structural changes in basement membranes at 1-3 days post fertilisation (dpf) were investigated. The perturbation observed across the inner limiting membranes may account for the lens dysplasia. Cell density of the granular epithelium in the photoreceptor cell layer was found to be lower in both morphants with the least density in fukutin knock-downs, which may result from a disruption of the external limiting membrane. This leads to the conclusion that fkrp and fukutin are essential for membrane integrity in the eye and muscle of developing zebrafish. A transgenic zebrafish line expressing enhanced green fluorescent protein (EGFP) in vascular endothelium from the fli-1 promoter was used to investigate early vascularisation. In all morphants, including dystroglycan knock downs, the intersegmental vessels failed to reach the dorsal longitudinal anastomotic vessel at 1dpf. Additionally, in the fkrp and fukutin morphant the eye vasculature was abnormal. Interestingly, no change was observed in the eye vasculature of the dystroglycan morphants suggesting that fkrp and fukutin may modify proteins other than α-dystroglycan in the eye.Medical Research Council as part of the MRC Centre for Neuromuscular Disease

MicroRNA-155 regulates monocyte chemokine and chemokine receptor expression in Rheumatoid Arthritis

Objectives: To test the hypothesis that miR-155 regulates monocyte migratory potential via modulation of chemokine and chemokine receptor expression in rheumatoid arthritis (RA); and thereby is associated with disease activity. Methods: miR-155 copy-number in monocytes from peripheral blood (PB) of healthy (n=22), RA (n=24), and RA synovial fluid (SF; n=11) were assessed by real time- PCR using synthetic miR-155 as quantitative standard. To evaluate the functional impact of miR-155, human monocytes were transfected with control or miR-155 mimic and the effect on transcript levels, and production of chemokines was evaluated by TLDA and multiplex assays. A comparative study evaluated constitutive chemokine receptor expression in miR-155-/- and wild-type murine (CD115+Ly6C+Ly6G-) monocytes. Results: Compared with healthy monocytes, miR-155 copy-number was higher in RA PB and SF monocytes (PB p<0.01, and SF p<0.0001). MiR-155 copy-number in RA PB monocytes were higher in ACPA positive compared with ACPA negative patients (p=0.033) and correlated (95% C.I.) with DAS28 (ESR), R=0.728 (0.460, 0.874), with tender, R=0.631 (0.306, 0.824) and swollen, R=0.503 (0.125, 0.753) joint counts. Enforced-expression of miR-155 in RA monocytes stimulated the production of CCL3, CCL4, CCL5, CCL8; up-regulated CCR7 expression and down-regulated CCR2. Conversely, miR155-/- monocytes showed down-regulated CCR7 and upregulated CCR2 expression. Conclusions: Given the observed correlations with disease activity, these data provide strong evidence that miR-155 can contribute to RA pathogenesis by regulating chemokine production and pro-inflammatory chemokine receptor expression, thereby promoting inflammatory cell recruitment and retention in the RA synovium

Terminal value: Building the alternative Bloomberg

In this essay we propose a reframing of the Bloomberg Terminal, an interface used to track financial trades and values, by using it as a creative, critical and curatorial device to explore the relationship between the art and finance. To contextualise this approach, we offer a history of the Bloomberg Terminal alongside an analysis of the power of interfaces to shape both the user and the represented information. We use the terminal as a way to critique the relationship between art acquisition and financial trading companies. We then describe some outcomes of a series of workshops themed around the idea of ‘building an alternative Bloomberg’. We conclude by offering some potential applications of a re-framed Bloomberg Terminal as an open and modular interface for engaging with issues around art and finance, both in terms of content and curation

Investigating Forest Photosynthetic Response to Elevated CO2 Using UAV-Based Measurements of Solar Induced Fluorescence

The response of ecosystems to increasing atmospheric CO2 will have significant, but still uncertain, impacts on the global carbon and water cycles. A lot of infounation has been gained from Free Air CO2 Enrichment (FACE) experiments, but the response of mature forest ecosystems remains a significant knowledge gap. One of the challenges in FACE studies is obtaining an integrated measure of canopy photosynthesis at the scale of the treatment ring. A new remote sensing approach for measuring photosynthetic activity is based on Solar Induced Fluorescence (SIF), which is emitted by plants during photosynthesis, and is closely linked to the rates and regulation of photosynthesis. We proposed that UAV-based SIF measurements, that enable the spectrometer field of view to be targeted to the treatment ring, provide a unique opportunity for investigating the dynamics of photosynthetic responses to elevated CO2. We have successfully tested this approach in a new FACE site, located in a mature oak forest in the UK. We flew a series of flights across the experiment arrays, collecting a number of spectra. We combined these with ground-based physiological and optical measurements, and see great promise in the use of UAV-based SIF measurements in FACE and other global change experiments.Peer reviewe

Participant Choice towards Receiving Potential Additional Findings in an Australian Nephrology Research Genomics Study

The choices of participants in nephrology research genomics studies about receiving additional findings (AFs) are unclear as are participant factors that might influence those choices. Methods: Participant choices and factors potentially impacting decisions about AFs were examined in an Australian study applying research genomic testing following uninformative diagnostic genetic testing for suspected monogenic kidney disease. Results: 93% of participants (195/210) chose to receive potential AFs. There were no statistically significant differences between those consenting to receive AFs or not in terms of gender (p = 0.97), median age (p = 0.56), being personally affected by the inherited kidney disease of interest (p = 0.38), or by the inheritance pattern (p = 0.12–0.19). Participants were more likely to choose not to receive AFs if the family proband presented in adulthood (p = 0.01), if there was family history of another genetic disorder (p = 0.01), and where the consent process was undertaken by an adult nephrologist (p = 0.01). Conclusion: The majority of participants in this nephrology research genomics study chose to receive potential AFs. Younger age of the family proband, family history of an alternate genetic disorder, and consenting by some multidisciplinary team members might impact upon participant choices

The HIDDEN Protocol: An Australian Prospective Cohort Study to Determine the Utility of Whole Genome Sequencing in Kidney Failure of Unknown Aetiology

Early identification of genetic kidney disease allows personalised management, clarification of risk for relatives, and guidance for family planning. Genetic disease is underdiagnosed, and recognition of genetic disease is particularly challenging in patients with kidney failure without distinguishing diagnostic features. To address this challenge, the primary aim of this study is to determine the proportion of genetic diagnoses amongst patients with kidney failure of unknown aetiology, using whole genome sequencing (WGS). A cohort of up to 100 Australian patients with kidney failure of unknown aetiology, with onset <50 years old and approved by a panel of study investigators will be recruited via 18 centres nationally. Clinically accredited WGS will be undertaken with analysis targeted to a priority list of ∼388 genes associated with genetic kidney disease. The primary outcome will be the proportion of patients who receive a molecular diagnosis (diagnostic rate) via WGS compared with usual -care (no further diagnostic investigation). Participant surveys will be undertaken at consent, after test result return and 1 year subsequently. Where there is no or an uncertain diagnosis, future research genomics will be considered to identify candidate genes and new pathogenic variants in known genes. All results will be relayed to participants via the recruiting clinician and/or kidney genetics clinic. The study is ethically approved (HREC/16/MH/251) with local site governance approvals in place. The future results of this study will be disseminated and inform practical understanding of the potential monogenic contribution to kidney failure of unknown aetiology. These findings are anticipated to impact clinical practice and healthcare policy. Study Registration: [https://dora.health.qld.gov.au], identifier [HREC/16/MH/251]

Responses to environmental enrichment differ with sex and genotype in a transgenic mouse model of Huntington's disease.

BACKGROUND: Environmental enrichment (EE) in laboratory animals improves neurological function and motor/cognitive performance, and is proposed as a strategy for treating neurodegenerative diseases. EE has been investigated in the R6/2 mouse model of Huntington's disease (HD), where increased social interaction, sensory stimulation, exploration, and physical activity improved survival. We have also shown previously that HD patients and R6/2 mice have disrupted circadian rhythms, treatment of which may improve cognition, general health, and survival. METHODOLOGY/PRINCIPAL FINDINGS: We examined the effects of EE on the behavioral phenotype and circadian activity of R6/2 mice. Our mice are typically housed in an "enriched" environment, so the EE that the mice received was in addition to these enhanced housing conditions. Mice were either kept in their home cages or exposed daily to the EE (a large playground box containing running wheels and other toys). The "home cage" and "playground" groups were subdivided into "handling" (stimulated throughout the experimental period) and "no-handling" groups. All mice were assessed for survival, body weight, and cognitive performance in the Morris water maze (MWM). Mice in the playground groups were more active throughout the enrichment period than home cage mice. Furthermore, R6/2 mice in the EE/no-handling groups had better survival than those in the home cage/no-handling groups. Sex differences were seen in response to EE. Handling was detrimental to R6/2 female mice, but EE increased the body weight of male R6/2 and WT mice in the handling group. EE combined with handling significantly improved MWM performance in female, but not male, R6/2 mice. CONCLUSIONS/SIGNIFICANCE: We show that even when mice are living in an enriched home cage, further EE had beneficial effects. However, the improvements in cognition and survival vary with sex and genotype. These results indicate that EE may improve the quality of life of HD patients, but we suggest that EE as a therapy should be tailored to individuals

Managing challenges in congenital CMV : current thinking

Congenital human cytomegalovirus (CMV) infection is the most common congenital infection, affecting around 1 in 200 infants in high-income settings. It can have life-long consequences for up to one in four children, including sensorineural hearing loss and neurodisability. Despite the frequency of congenital CMV and the severity for some children, it is a little-known condition by pregnant women, families and healthcare providers. Timely diagnosis of CMV infection in pregnancy is important to facilitate consideration of treatment with valaciclovir, which may reduce the risk of transmission to the fetus or reduce the severity of the outcomes for infected infants. Recognition of features of congenital CMV is important for neonatologists, paediatricians and audiologists to prompt testing for congenital CMV within the first 21 days of life. Early diagnosis gives the opportunity for valganciclovir treatment, where appropriate, to improve outcomes for affected infants. Further research is urgently needed to inform decisions about antenatal and neonatal screening, long-term outcomes for asymptomatic and symptomatic infants, predictors of these outcomes and optimal treatment for women and infants

Royal succession and kingship among the Picts

When we consider the history of the Picts we are faced with the perennial challenge for the early medievalist of deciding whether the fragments of evidence which survive are representative of the reality of Pictish society, or whether they provide us with distortions, based on patterns of survival. This issue is as relevant to the subject of royal succession as it is to other aspects of Pictish history. The debate over whether the Picts practised a matrilineal system, with the son of the previous king's sister becoming the next king, or whether it was a patrilineal system, with the kingship generally passing through the male line, has dominated the discussion of Pictish succession. Until the 1980s, the matriliny thesis was virtually unquestioned, and accepted by scholars including F. T. Wainwright, Marjorie Anderson, and Isabel Henderson1. The bases for this view were the accounts of the Pictish settlement of northern Britain in Bede's ‘Ecclesiastical History of the English People’ and Irish texts written throughout the medieval period, but mainly surviving in versions from the twelfth century or later.2 In these sources it was claimed that the Picts went to Ireland before arriving in northern Britain, and that they obtained wives from the Irish, with some versions stating that this was done on condition that the succession went through the female line. Other sources which did not openly discuss the nature of Pictish succession, particularly the Irish chronicles and the Pictish king-lists, were then interpreted by scholars in relation to these accounts and were thought to support them

Sustained Reduction in Third-generation Cephalosporin Usage in Adult Inpatients Following Introduction of an Antimicrobial Stewardship Program in a Large, Urban Hospital in Malawi

Background Third-generation cephalosporins (3GC) remain the first-choice empiric antibiotic for severe infection in many sub-Saharan African hospitals. In Malawi, limited availability of alternatives, mean that strategies to prevent spread of 3GC-resistance (3GC-R) are imperative, however suitable approaches to antimicrobial stewardship (AMS) in low-income settings are not well studied. Methods We introduced an AMS intervention to Queen Elizabeth Central Hospital (QECH) in Blantyre. The intervention consisted of a smartphone prescribing application and regular point-prevalence surveys (PPS) with prescriber feedback. We evaluate the effects of the intervention on 3GC usage and on cost of providing antibiotics. Using thematic analysis of semi-structured interviews and participant observations, we additionally evaluate the acceptability of the stewardship program. Results The proportion of antibiotic prescriptions for a 3GC reduced from 193/241 (80.1%) to 177/330 (53.6%) (percentage decrease 26.5% [95%CI; 18.7 to 34.1]) with no change in case-fatality rate. Cost analysis estimated annual savings of US$15,000. Qualitative research revealed trust in the guideline and found its accessibility through smartphones helpful to guide clinical decisions. Operational health-system barriers and hierarchal clinical relationships lead to continued reliance on 3GC. Conclusions We report the successful introduction of an antimicrobial stewardship approach in Malawi. By focusing on pragmatic interventions and simple aims, we demonstrate the feasibility, acceptability and cost-saving of a stewardship program where resources are limited. In doing so, we provide a suitable starting point for expansion of AMS interventions in this and other low-income settings