The CD4+ T cell methylome contributes to a distinct CD4+ T cell transcriptional signature in Mycobacterium bovis-infected cattle

peer-reviewedWe hypothesised that epigenetic regulation of CD4+ T lymphocytes contributes to a shift toward a dysfunctional T cell phenotype which may impact on their ability to clear mycobacterial infection. Combined RNA-seq transcriptomic profiling and Reduced Representation Bisulfite Sequencing identified 193 significantly differentially expressed genes and 760 differentially methylated regions (DMRs), between CD4+ T cells from M. bovis infected and healthy cattle. 196 DMRs were located within 10 kb of annotated genes, including GATA3 and RORC, both of which encode transcription factors that promote TH2 and TH17 T helper cell subsets respectively. Gene-specific DNA methylation and gene expression levels for the TNFRSF4 and Interferon-γ genes were significantly negatively correlated suggesting a regulatory relationship. Pathway analysis of DMRs identified enrichment of genes involved in the anti-proliferative TGF-β signaling pathway and TGFB1 expression was significantly increased in peripheral blood leukocytes from TB-infected cattle. This first analysis of the bovine CD4+ T cell methylome suggests that DNA methylation directly contributes to a distinct gene expression signature in CD4+ T cells from cattle infected with M. bovis. Specific methylation changes proximal to key inflammatory gene loci may be critical to the emergence of a non-protective CD4+ T cell response during mycobacterial infection in cattle

Prevalence of Non-Prescribed Drug Use in Hospital Patients Assessed by Urine Toxicology Testing

To analyse the results of a survey for non-prescribed drug use in selected patient populations in Tallaght University Hospital to determine the patterns of drug use by urine toxicology testing. Urine toxicology screening results done by the Alere Triage® TOX Drug Screen Meter were extracted from the Clinical Chemistry Lab database from the 5th of March to the 23rd of March at Tallaght University Hospital. Results were analysed to determine which drug tested positive most commonly. Benzodiazepines were the most prevalent drugs of abuse in urine toxicology, accounting for 25.62% of all positive results, this was followed by Cannabis and Amphetamines with 21.67% and 20.20% respectively. The largest age group that presented was between 30-39 inclusive. Benzodiazepines are the most prevalent positive result in drug of abuse screens in Tallaght University Hospital and the 30-39 age group contained the most positives and number of samples sent for toxicology analysis, supporting the claims of recent literature

Circumventing the "Ick" factor: A randomized trial of the effects of omitting affective attitudes questions to increase intention to become an organ donor

Objectives: Including or excluding certain questions about organ donation may influence peoples’ intention to donate. We investigated the effect of omitting certain affective attitudinal items on potential donors’ intention and behavior for donation.  Design: A cross-sectional survey with a subgroup nested randomized trial.  Methods: A total of 578 members of the public in four shopping centers were surveyed on their attitudes to organ donation. Non-donors (n= 349) were randomly assigned to one of three groups: Group 1 completed items on affective and cognitive attitudes, anticipated regret, intention, subjective norm and perceived behavioral control. Group 2 completed all items above but excluded affective attitudes. Group 3 completed all items but omitted negatively worded affective attitudes. The primary outcome was intention to donate, taking a donor card after the interview was a secondary behavioral outcome, and both were predicted using linear and logistic regression with group 1 as the reference.  Results: Mean (SD) 1–7 intention scores for groups 1, 2 and 3 were, respectively: 4.43 (SD 1.89), 4.95 (SD 1.64) and 4.88 (SD 1.81), with group 2 significantly higher than group 1 (β = 0.518, 95% confidence interval [CI] 0.18 to 0.86).At the end of the interview, people in group 2 (66.7%; OR = 1.40, 95% CI 0.94 to 2.07,p= 0.096) but not those in group 3 (61.7%; OR = 1.10, 95% CI 0.69 to 1.75,p= 0.685), were marginally more likely to accept a donor card from the interviewer than people in group 1 (59.7%).  Conclusion: Omitting affective attitudinal items results in higher intention to donate organs and marginally higher rates of acceptance of donor cards, which has important implications for future organ donation public health campaigns

On the modelling and closed loop control of an inductively coupled plasma chamber

As a first step towards real time, multivariable control of an argon/ oxygen plasma, the implementation of real time control of ion flux in an inductively coupled argon plasma through modulation of the RF power is described. It is demonstrated that an elementary PID controller does not guarantee stable control of ion flux over a range of operating points and hence that more elaborate control strategies must be considered. The design and testing of control algorithms is facilitated by suitable dynamical models of a process. A model of the inductively coupled plasma chamber which is suitable for control simulations is described. Ongoing and future work are discussed

Theory Content, Question-Behavior Effects, or Form of Delivery Effects for Intention to Become an Organ Donor? Two Randomized Trials

Eliciting different attitudes with survey questionnaires may impact on intention to donate organs. Previous research used varying numbers of questionnaire items, or different modes of intervention delivery, when comparing groups. We aimed to determine whether intention to donate organs differed among groups exposed to different theoretical content, but similar questionnaire length, in different countries. We tested the effect of excluding affective attitudinal items on intention to donate, using constant item numbers in two modes of intervention delivery. Study 1: A multi-country, interviewer-led, cross-sectional randomized trial recruited 1007 participants, who completed questionnaires as per group assignment: including all affective attitude items, affective attitude items replaced, negatively-worded affective attitude items replaced. Study 2 recruited a UK-representative, cross-sectional sample of 616 participants using an online methodology, randomly assigned to the same conditions. Multilevel models assessed effects of group membership on outcomes: intention to donate (primary), taking a donor card, following a web-link (secondary). In study 1, intention to donate did not differ among groups. Study 2 found a small, significantly higher intention to donate in the negatively-worded affective attitudes replaced group. Combining data yielded no group differences. No differences were seen for secondary outcomes. Ancillary analyses suggest significant interviewer effects. Contrary to previous research, theoretical content may be less relevant than number or valence of questionnaire items, or form of intervention delivery, for increasing intention to donate organs

Stretching the IR theoretical spectrum on Irish neutrality: a critical social constructivist framework

In a 2006 International Political Science Review article, entitled "Choosing to Go It Alone: Irish Neutrality in Theoretical and Comparative Perspective," Neal G. Jesse argues that Irish neutrality is best understood through a neoliberal rather than a neorealist international relations theory framework. This article posits an alternative "critical social constructivist" framework for understanding Irish neutrality. The first part of the article considers the differences between neoliberalism and social constructivism and argues why critical social constructivism's emphasis on beliefs, identity, and the agency of the public in foreign policy are key factors explaining Irish neutrality today. Using public opinion data, the second part of the article tests whether national identity, independence, ethnocentrism, attitudes to Northern Ireland, and efficacy are factors driving public support for Irish neutrality. The results show that public attitudes to Irish neutrality are structured along the dimensions of independence and identity, indicating empirical support for a critical social constructivist framework of understanding of Irish neutrality

Assessing medical mistrust in organ donation across countries using item response theory

Although medical mistrust (MM) may be an impediment to public health interventions, no MM scale has been validated across countries and the assessment of MM has not been explored using item response theory, which allows generalisation beyond the sampled data. We aimed to determine the dimensionality of a brief MM measure across four countries through Mokken analysis and Graded Response Modelling. Analysis of 1468 participants from UK (n = 1179), Ireland (n = 191), India (n = 49) and Malaysia (n = 49) demonstrated that MM items formed a hierarchical, unidimensional measure, which is very informative about high levels of MM. Possible item reduction and scoring changes were also demonstrated. This study demonstrates that this brief MM measure is suitable for international studies as it is unidimensional across countries, cross cultural, and shows that minor adjustments will not impact on the assessment of MM when using these items.Output Status: Forthcoming/Available Onlin

Heritability of subcortical volumetric traits in mesial temporal lobe epilepsy.

OBJECTIVES: We aimed to 1) determine if subcortical volume deficits are common to mesial temporal lobe epilepsy (MTLE) patients and their unaffected siblings 2) assess the suitability of subcortical volumetric traits as endophenotypes for MTLE. METHODS: MRI-based volume measurements of the hippocampus, amygdala, thalamus, caudate, putamen and pallidium were generated using an automated brain reconstruction method (FreeSurfer) for 101 unrelated 'sporadic' MTLE patients [70 with hippocampal sclerosis (MTLE+HS), 31 with MRI-negative TLE], 83 unaffected full siblings of patients and 86 healthy control subjects. Changes in the volume of subcortical structures in patients and their unaffected siblings were determined by comparison with healthy controls. Narrow sense heritability was estimated ipsilateral and contralateral to the side of seizure activity. RESULTS: MTLE+HS patients displayed significant volume deficits across the hippocampus, amygdala and thalamus ipsilaterally. In addition, volume loss was detected in the putamen bilaterally. These volume deficits were not present in the unaffected siblings of MTLE+HS patients. Ipsilaterally, the heritability estimates were dramatically reduced for the volume of the hippocampus, thalamus and putamen but remained in the expected range for the amygdala. MRI-negative TLE patients and their unaffected siblings showed no significant volume changes across the same structures and heritability estimates were comparable with calculations from a healthy population. CONCLUSIONS: The findings indicate that volume deficits for many subcortical structures in 'sporadic' MTLE+HS are not heritable and likely related to acquired factors. Therefore, they do not represent suitable endophenotypes for MTLE+HS. The findings also support the view that, at a neuroanatomical level, MTLE+HS and MRI-negative TLE represent two distinct forms of MTLE

Medical student experiences of Case-Based Learning (CBL) at a multicultural medical school

Educational research highlights active approaches to learning are more efective in knowledge retention and problem-solving. It has long been acknowledged that adapting to more active ways of learning form part of the challenge for new university students as the pedagogical distance between the didactical approach largely followed by secondary school systems the world over difers quite signifcantly from the often more student-led, critical approach taken by universities. University students encounter various learning challenges, particularly during the transition from secondary school to university. Poor adaptation and low performance in the frst year of tertiary education can lead to higher failure rates and potential withdrawal from study programmes. Adopting active learning strategies early in this transition phase is crucial for supporting students’ adaptation and success. Gaining student engagement with active learning can be a signifcant challenge when there is an expectation to participate in a discussion or voice an opinion. Case-based learning (CBL), with its scafolded form of learning, is an approach that could provide the support needed to help multicultural learners adapt to their new learning environment in a non-threatening classroom-based setting. The research question in this study was: what features of CBL support active learning

Somatic variants as a cause of drug-resistant epilepsy including mesial temporal lobe epilepsy with hippocampal sclerosis

OBJECTIVE: The contribution of somatic variants to epilepsy has recently been demonstrated, particularly in the etiology of malformations of cortical development. The aim of this study was to determine the diagnostic yield of somatic variants in genes that have been previously associated with a somatic or germline epilepsy model, ascertained from resected brain tissue from patients with multidrug-resistant focal epilepsy. METHODS: Forty-two patients were recruited across three categories: (1) malformations of cortical development, (2) mesial temporal lobe epilepsy with hippocampal sclerosis, and (3) nonlesional focal epilepsy. Participants were subdivided based on histopathology of the resected brain. Paired blood- and brain-derived DNA samples were sequenced using high-coverage targeted next generation sequencing to high depth (585— and 1360—, respectively). Variants were identified using Genome Analysis ToolKit (GATK4) MuTect-2 and confirmed using high-coverage Amplicon-EZ sequencing. RESULTS: Sequence data on 41 patients passed quality control. Four somatic variants were validated following amplicon sequencing: within CBL, ALG13, MTOR, and FLNA. The diagnostic yield across 41 patients was 10%, 9% in mesial temporal lobe epilepsy with hippocampal sclerosis and 20% in malformations of cortical development. SIGNIFICANCE: This study provides novel insights into the etiology of mesial temporal lobe epilepsy with hippocampal sclerosis, highlighting a potential pathogenic role of somatic variants in CBL and ALG13. We also report candidate diagnostic somatic variants in FLNA in focal cortical dysplasia, while providing further insight into the importance of MTOR and related genes in focal cortical dysplasia. This work demonstrates the potential molecular diagnostic value of variants in both germline and somatic epilepsy genes