Foundation Center Research Articles

Future Leaders to Watch – Christopher Toepfer and Manuel Schmid

ABSTRACTFuture Leaders to Watch is a series of interviews with the first authors of a selection of papers published in Biology Open, helping early-career researchers promote themselves alongside their papers. Christopher Toepfer and Manuel Schmid are co-first authors on ‘Myosin super relaxation (SRX) a perspective on fundamental biology, human disease, and cardiac therapeutics’, published in BiO. Christopher is a Sir Henry Wellcome Postdoctoral Fellow and British Heart Foundation Centre of Research Excellence (CRE) Intermediate Transition Fellow in the department of Cardiovascular Medicine at the Radcliffe Department of Medicine, University of Oxford, UK. Manuel is a Research Assistant in the department of Cardiovascular Medicine at the Radcliffe Department of Medicine, University of Oxford, UK. They are investigating the mechanisms that control heart function and impact acquired and inherited cardiovascular conditions.

Texture analysis of ultra-high coercivity Sm2Co7 hot deformation magnets* *Project supports by the Science Center of the National Natural Science Foundation of China (Grant No. 52088101), the National Natural Foundation of China (Grant No. 51590880), the Fujian Institute of Innovation, Chinese Academy of Sciences (Grant No. FJCXY18040302), the Key Program of the Chinese Academy of Sciences (Grant No. KJZD-EW-M05-1), and

Bulk anisotropic Sm2Co7 nanocrystalline magnets were successfully prepared by hot deformation process using spark plasma sintering technology. The coercivity of the isotropic Sm2Co7 nanocrystalline magnet is 34.76 kOe, further, the ultra-high coercivity of 50.68 kOe is obtained in the anisotropic hot deformed Sm2Co7 magnet when the height reduction is 70%, which is much higher than those of the ordinarily produced hot deformed Sm2Co7 magnet. X-ray diffraction (XRD) analysis shows that all the samples are Sm2Co7 single phase. The investigation by electron backscatter diffraction indicates that increasing the amount of deformation is beneficial to the improvement of the (00l) texture of Sm2Co7 magnets. The Sm2Co7 nanocrystalline magnet generates a strong c-axis crystallographic texture during large deformation process.

Amalgamation of self-esteem, depression, anxiety and stress among prosthesis users

To find the levels of self-esteem, depression, anxiety and stress among prosthesis users. The cross-sectional study was conducted from June to November 2018 at Chal Foundation centres in Bagh and Swabi, Pakistan, and comprised prosthesis users of both genders aged 15-60 years. Rosenberg self-esteem scale and Depression anxiety and stress scale-21 were used to collect data which was analysed using SPSS 22. Of the 400 subjects, 315(78.8%) were males and 85(21.2%) were females. The overall mean age was 38.03±11.86 years. Low level of self-esteem was found in 350(87.5%), depression in 374(93.4%), anxiety in 388(96.9%) and stress in 338(84.4%). Females showed significant association with stress (p=0.009). The level of self-esteem was low among majority of the participants and psychological well-being was found to be poor.

Decade-Long Trends in Surgery for Acute Type A Aortic Dissection

Background: Little is known about unwarranted variations in care, with regional differences and outcomes of patients who undergo surgical repair for type A acute aortic dissection (TAAD) in England. We aim to investigate decade-long trends in TAAD surgical repair in England using national-level data. Method: Retrospective review of the National Institute for Cardiovascular Outcomes Research (NICOR) National Adult Cardiac Surgery Audit (NACSA) registry, which prospectively collects demographic, as well as pre-, peri- and post-operative clinical information, including mortality, for all major adult cardiac surgery procedures performed in the UK. We identified patients undergoing surgery for TAAD from January 2009 to December 2018 and reviewed trends in operative frequency and mortality. Findings: Over the 10-year period,3,686 TAAD patients underwent surgical repair in England. A steady doubling in the overall number of operations conducted in England was observed from 237 cases recorded in 2009 to 510 in 2018. Number of procedures per hospital per year also doubled, from 10 in 2009 to 21 in 2018. The risk profile of the operated patients remained unchanged. Overall, in-hospital mortality was 17.4% with a trend toward lower mortality in the most recent years (from 22.8% in 2009 to 14.7% in 2018). There was a significant variation in operative mortality across regions with a trend towards lower mortality in regions with a high-volume hospital. Interpretation: Surgery is the only treatment for acute TAAD but is associated with high mortality. Prompt diagnosis and referral to a specialist center is paramount. The number of operations conducted in England has doubled in 10 years and the associated survival following surgery has improved. Regional variations exist in service provision with a trend towards better survival in high volume centers. Funding Statement: British Heart Foundation and NIHR Biomedical Research Centre at University Hospitals Bristol and Weston NHS Foundation Trust and University of Bristol. Declaration of Interests: Authors have no conflict of interests to declare. Ethics Approval Statement: This study is part of research approved by the Health Research Authority (HRA) and Health and Care Research Wales (HCRW) (IRAS ID: 278171).

Modified lysmer’s analog model for two dimensional mat settlements under vertically uniform load

A two dimensional model of linearly elastic soil spring used for the settlement analysis of the flexible mat foundation is suggested in this study. The spring constants of the soils underneath the foundation were modeled assuming uniformly vertical load applied onto the foundation. The soil spring constants were back calculated using the three-dimensional finite element analysis with Midas GTS NX program. Variation of the soil spring constants was modeled as a two-dimensional polynomial function in terms of the normalized spatial distances between the center of foundation and the analytical points. The Lysmer's analog spring for soils underneath the rigid foundation was adopted and calibrated for the flexible foundation. For validations, the newly proposed soil spring model was incorporated into a two dimensional finite difference analysis for a square mat foundation at the surface of an elastic half-space consisting of soft clays. Comparative study was made for elastic soils where the shear wave velocity is 120~180 m/s and the Poisson's ratio varies at 0.3~0.5. The resulting foundation settlements from the two dimensional finite difference analysis with the proposed soil springs were found in good agreement with those obtained directly from three dimensional finite element analyses. Details of the applications and limitations of the modified Lysmer's analog springs were discussed in this study.

Economic crisis in mass representations of Russians

During the post-Soviet period, Russians have experienced several economic crises, which have become a prism for assessing the political situation in the country and the direction of its development. In this article, based on mass polls data by the Public Opinion Foundation (FOM) and Russian Public Opinion Research Center (VCIOM) in 2019-2020, the author has reconstructed public perceptions of the economic crisis, identified the groups more inclined to admit the crisis in the country, described adaptive mechanisms they used, and assessed if the feeling of crisis affect the public trust in political institutions. A quantitative textual analysis of the open entry responses concerning the characteristics of the economic crisis shows that it is consistently associated with the gap between available and desired levels of consumption and economic security, rather than a sharp decline in macroeconomic indicators. Rising prices, low living standards and low wages are the most frequent indicators of the crisis in the responses. Respondents in the low-income categories and younger cohorts are the most susceptible to the crisis. Adaptation mechanisms had already begun to form on the eve of the main wave of the pandemic in March 2020, but the increased level of uncertainty about the future determined the demand for trustful information on the state of affairs in the economy. The study also shows that the feeling of crisis is associated with the decreased distrust of governmental bodies. In general, the work demonstrates the importance of studying subjective assessments of well-being and their role in structuring information about processes in politics and economy.

Medium-term effects of SARS-CoV-2 infection on multiple vital organs, exercise capacity, cognition, quality of life and mental health, post-hospital discharge.

BackgroundThe medium-term effects of Coronavirus disease (COVID-19) on organ health, exercise capacity, cognition, quality of life and mental health are poorly understood.MethodsFifty-eight COVID-19 patients post-hospital discharge and 30 age, sex, body mass index comorbidity-matched controls were enrolled for multiorgan (brain, lungs, heart, liver and kidneys) magnetic resonance imaging (MRI), spirometry, six-minute walk test, cardiopulmonary exercise test (CPET), quality of life, cognitive and mental health assessments.FindingsAt 2–3 months from disease-onset, 64% of patients experienced breathlessness and 55% reported fatigue. On MRI, abnormalities were seen in lungs (60%), heart (26%), liver (10%) and kidneys (29%). Patients exhibited changes in the thalamus, posterior thalamic radiations and sagittal stratum on brain MRI and demonstrated impaired cognitive performance, specifically in the executive and visuospatial domains. Exercise tolerance (maximal oxygen consumption and ventilatory efficiency on CPET) and six-minute walk distance were significantly reduced. The extent of extra-pulmonary MRI abnormalities and exercise intolerance correlated with serum markers of inflammation and acute illness severity. Patients had a higher burden of self-reported symptoms of depression and experienced significant impairment in all domains of quality of life compared to controls (p<0.0001 to 0.044).InterpretationA significant proportion of patients discharged from hospital reported symptoms of breathlessness, fatigue, depression and had limited exercise capacity. Persistent lung and extra-pulmonary organ MRI findings are common in patients and linked to inflammation and severity of acute illness.FundingNIHR Oxford and Oxford Health Biomedical Research Centres, British Heart Foundation Centre for Research Excellence, UKRI, Wellcome Trust, British Heart Foundation.

How Should We Sequence the Treatments for Heart Failure and a Reduced Ejection Fraction?: A Redefinition of Evidence-Based Medicine.

How Should We Sequence the Treatments for Heart Failure and a Reduced Ejection Fraction?: A Redefinition of Evidence-Based Medicine.

Understanding the Relationship Between Students’ Mathematics Anxiety Levels and Mathematics Performances at the Foundation Level

For many students, mathematics is regarded as a challenging subject to learn and master in class. One of the significant factors contributing to the students&amp;rsquo; difficulties in learning mathematics is caused by a phenomenon called mathematics anxiety. Mathematics anxiety is a feeling of unease and anxiety toward mathematics and it can be different from person-to-person. Understanding the effects of mathematics anxiety levels on students&amp;rsquo; mathematics performances in class can be the key to help students&amp;rsquo; mastery of mathematics. The aim of the study is to investigate the relationship between mathematics anxiety levels and students&amp;rsquo; mathematics performances at the foundation level. A sample of 545 students from a local foundation centre was chosen for this study. Data collection via questionnaire was used where quantitative data were analysed using correlation, t-test, and descriptive analyses. The results showed that there was a weak positive correlation between students&amp;rsquo; anxiety levels and the students&amp;rsquo; mathematics performance in their final examination. Recommendations and future potential for this study were further discussed in this paper.

SARS-CoV-2 in first trimester pregnancy: a cohort study.

Study questionDoes maternal infection with SARS-CoV-2 in first trimester pregnancy have an impact on the fetal development as measured by nuchal translucency thickness and pregnancy loss?Summary answerNuchal translucency thickness at the first trimester scan was not significantly different in pregnant women with versus without SARS-CoV-2 infection in early pregnancy and there was no significant increased risk of pregnancy loss in women with SARS-CoV-2 infection in the first trimester.What is known alreadyPregnant women are more vulnerable to viral infections. Previous coronavirus epidemics have been associated with increased maternal morbidity, mortality and adverse obstetric outcomes. Currently, no evidence exists regarding possible effects of SARS-CoV-2 in first trimester pregnancies.Study design, size, durationCohort study of 1,019 women with a double test taken between Feb. 17 and Apr. 23, 2020, as a part of the combined first trimester risk assessment, and 36 women with a first trimester pregnancy loss between Apr. 14 and May 21, 2020, prior to the double test. The study period was during the first SARS-CoV-2 epidemic wave in Denmark.Participants/materials, setting, methodsCohort 1 included pregnant women with a double test taken within the study period. The excess serum from each double test was analyzed for SARS-CoV-2 antibodies. Results were correlated to the nuchal translucency thickness and the number of pregnancy losses before or at the time of the first trimester scan. Cohort 2 included women with a pregnancy loss before the gestational age for double test sample. Serum from a blood test taken the day the pregnancy loss was identified was analyzed for SARS-CoV-2 antibodies. The study was conducted at a public university hospital serving approximately 12% of pregnant women and births in Denmark. All participants in the study provided written informed consent.Main results and the role of chanceEighteen (1.8%) women had SARS-CoV-2 antibodies in the serum from the double test suggestive of SARS-CoV-2 infection in early pregnancy. There was no significant difference in nuchal translucency thickness for women testing positive for previous SARS-CoV-2 infection (n = 18) versus negative (n = 994) (p = 0.62). There was no significant increased risk of pregnancy loss for women with positive antibodies (n = 1) (OR 3.4, 0.08-24.3 95% CI, p = 0.27). None of the women had been hospitalized due to SARS-CoV-2 infection. None of the women with pregnancy loss prior to the double test (Cohort 2) had SARS-CoV-2 antibodies.Limitations, reasons for cautionThese results may only apply to similar populations and to patients who do not require hospitalization due to SARS-CoV-2 infection. A limitation of the study is that only 1.8% of the study population had SARS-CoV-2 antibodies suggestive of previous infection.Wider implication of the findingsMaternal SARS-CoV-2 infection had no effect on the nuchal translucency thickness and there was no significant increased risk of pregnancy loss for women with SARS-CoV-2 infection in first trimester pregnancy. Evidence concerning Covid-19 in pregnancy is still limited. These data indicate that infection with SARS-CoV-2 in not hospitalized women does not pose a significant threat in first trimester pregnancies. Follow up studies are needed to establish any risk to a fetus exposed to maternal SARS-CoV-2 infection.Study funding/competing interest(s)Prof. Henriette Svarre Nielsen (HSN) and colleagues received a grant from the Danish Government for research of Covid-19 among pregnant women. The Danish government was not involved in the study design, data collection, analysis, interpretation of data, writing of the report or decision to submit the paper for publication. AI, JOL, JBR, DMS, JEF, and ERH received funding from a Novo Nordisk Foundation (NNF) Young Investigator Grant (NNF15OC0016662) and a Danish National Science Foundation Center Grant (6110-00344B). AI received a Novo Scholarship. JOL is funded by an NNF Pregraduate Fellowship (NNF19OC0058982). DW is funded by the NNF (NNF18SA0034956, NNF14CC0001, NNF17OC0027594). AMK is funded by a grant from the Rigshospitalet’s research fund. Henriette Svarre Nielsen has received speakeŕs fees from Ferring Pharmaceuticals, Merck Denmark A/S and Ibsa Nordic (outside the submitted work). Nina la Cour Freiesleben has received a grant from Gedeon Richter (outside the submitted work). Astrid Marie Kolte has received speakeŕs from Merck (outside the submitted work). The other authors did not report any potential conflicts of interest.

Game-based self-regulated language learning: Theoretical analysis and bibliometrics.

Game-based learning and self-regulated learning have long been valued as effective approaches to language education. However, little research has been conducted to investigate their integration, namely, game-based self-regulated language learning (GBSRLL). This study aims to conceptualise GBSRLL based on the combination of theoretical analysis, thematic evolution analysis, and social network analysis on the research articles in the fields of game-based language learning and self-regulated language learning. The results show that GBSRLL is a new interdisciplinary field emerging since the period from 2018 to 2019. Self-regulated learning strategies that can be performed in GBSRLL, the effects of GBSRLL on learners’ affective states, and the features in GBSRLL were the prominent research topics in this field. Its theoretical foundation centres on the positive correlations between learner motivation, self-efficacy, and autonomy and the implementation of game-based learning and self-regulated learning. It is feasible to conduct GBSRLL due to the strong supportiveness of game mechanics for various phases and strategies of self-regulated learning. More contributions to this new interdisciplinary field are called for, especially from the aspects of the long-term effects of GBSRLL on academic performance and the useful tools and technologies for implementing GBSRLL.

AJRH gets new numbers and a social science editor.

AJRH gets new numbers and a social science editor.

Prevalence and Risk Factors of Hypovitaminosis D in Nigerian Children with Sickle Cell Anaemia

Background: Vitamin D deficiency (VDD) has been linked to some acute and chronic bone disorders that commonly complicate sickle cell anaemia (SCA) in children. Some of these bone diseases include chronic pain, reduced bone density and fractures. Despite Nigeria having the highest number of children with SCA in the world, there is a paucity of data on vitamin D status and the associated risk factors in affected children. Objective: To determine the prevalence and risk factors for hypovitaminosis D in children with sickle cell anaemia in steady-state. Methods: A total of 174 children with sickle cell anaemia aged one to eighteen years were recruited at the Sickle Cell Foundation Centre, Lagos. Baseline sociodemographic, clinical, anthropometric and laboratory parameters (serum 25-hydroxyvitamin D, corrected serum calcium and alkaline phosphatase) were recorded. Results: The prevalence of vitamin D insufficiency and deficiency were 12.6 % and 72.5% respectively. Children below six years of age were less likely to have hypovitaminosis D compared to the older age groups (p = 0.017). The mean serum corrected calcium was lowest in subjects with vitamin D deficiency (p &gt;0.001). Age and hypocalcaemia are independent predictors of hypovitaminosis D. Conclusion: There is a high prevalence of vitamin D deficiency among children with sickle cell anaemia. Children aged below six years and with those with hypocalcaemia had higher odds of hypovitaminosis D.

An optimised failure envelope approach of bucket foundation in undrained clay

ABSTRACT The bucket foundation is a new type foundation which is suitable for offshore wind turbines. The failure envelope is used to check the bearing capacity under combined loading conditions, especially for foundations with irregular shapes. Due to the coupling effect between horizontal and moment loads, the H-M failure envelope has an obvious asymmetry, which makes the established algebraic equation of the envelope form complex and inconvenient to apply. In this paper, an optimised failure envelope is proposed to reduce the coupling degree between horizontal and moment loads by moving the load reference point to the rotation centre of bucket foundation. Furthermore, the algebraic equation of the failure envelope is established based on the optimised envelope by using the depth of the bucket foundation rotation centre. The equation has fewer parameters and clearer physical meanings than the conventional one, which is convenient for engineering applications.

In-vivo validation of non-invasive electrocardiographic imaging based on the equivalent double layer source model using epicardial and endocardial electroanatomical mapping

Abstract Background Noninvasive electrocardiographic imaging (ECGi) using the equivalent double layer (EDL) source model enables both epicardial and endocardial reconstruction of electroanatomical activation patterns during sinus rhythm. The EDL source model has been validated in torso-tank models and animal experiments, however validation in humans using invasive electroanatomical mapping is limited. Purpose Validation of EDL based ECGi using invasive electroanatomical mapping. Methods Ten patients referred for epicardial and endocardial electroanatomical mapping underwent 67 electrode body surface potential mapping (BSPM), cardiac CT imaging and 3D imaging of electrode positions. Anatomical models of the ventricles, lungs and thorax were created and supplemented with electrode positions. Invasive epicardial (4020±1514 contact points), right ventricle endocardial (724±113 contact points) and left ventricle endocardial (459±117 contact points) local activation timing (LAT) maps were compared to ECGi derived LAT maps. Results Included patients (mean age 48±21 years, 80% males) were diagnosed with either arrhythmogenic cardiomyopathy (N=4), dilated cardiomyopathy (N=1), symptomatic premature ventricular complexes (N=3) or myocarditis (N=2). Measured BSMP highly correlated with simulated BSPM (97±2%) with a relative difference of 24% ± 6%. Source models consisted of a mean of 2614±214 nodes with a mean distance between nodes of 8.3±1 mm. Overall, invasive LAT maps and ECGi derived LAT maps showed reasonable correlation for epicardial maps (49% ± 18%) and endocardial maps of the right ventricle (48% ± 27%). Endocardial left ventricular LAT maps showed less correlation (23% ± 28). The absolute difference between invasive and ECGi LAT maps was 16.7±14.2 ms for epicardial maps, 17.2±12.9 ms for right ventricle endocardial maps and 34.0±19.8 ms for left ventricle endocardial maps. Visual comparison showed corresponding areas of breakthrough and regions of latest activation in all cases (Figure). Quantitative comparison of these areas showed a mean absolute difference of 33±7mm or 19±10 ms for epicardial maps, 25±10 mm or 17±13 ms for right ventricle endocardial maps and 39±16 mm or 20±6 ms for left ventricular endocardial maps. Conclusion Overall agreement was observed between EDL based ECGi LAT maps and electroanatomical LAT maps. Future research will focus on further quantification of this agreement and improvement of ECGi mapping accuracy. Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): Netherlands Cardiovascular Research Initiative, an initiative with support of the Dutch Heart Foundation and UCL Hospitals NIHR Biomedical Research Centre.

Abnormalities of the lymphatic system and impaired fluid clearance in patients with heart failure with preserved ejection fraction

Abstract Background Coronary and skeletal muscle microvascular dysfunction have been proposed as main factors in the pathogenesis of Heart Failure with Preserved Ejection Fraction (HFpEF). However, assessment of systemic arterial function has only been indirect thus far; most importantly, no direct link between systemic microvasculature and congestion, one of the core characteristics of the syndrome, has yet been investigated. Purpose To provide direct functional and anatomical characterisation of the systemic microvasculature and to explore in vivo parameters of capillary fluid extravasation and lymphatic clearance in HFpEF. Methods In 16 patients with HFpEF and 16 age- and sex-matched healthy controls (72±6 and 68±5 years, respectively) we determined peripheral microvascular filtration coefficient (proportional to vascular permeability and area) and isovolumetric pressure (above which lymphatic drainage cannot compensate for fluid extravasation) by venous occlusion plethysmography and collected a skin biopsy for vascular immunohistochemistry and gene expression analysis (TaqMan). Additionally, we measured brachial flow-mediated dilatation (FMD) and assessed by wire myography the vascular function of resistance arteries isolated from gluteal subcutaneous fat biopsies. Results Skin biopsies in patients with HFpEF showed rarefaction of small blood vessels (82±31 vs 112±21 vessels/mm2; p=0.003) and in ex-vivo analysis (n=6/group) we found defective relaxation of peripheral resistance arteries (p&amp;lt;0.001). Accordingly, post-ischaemic hyperaemic response (fold-change vs baseline, 4.6±1.6 vs 6.7±1.7; p=0.002) and FMD (3.9±2.1 vs 5.6±1.5%; p=0.014) were found to be reduced in patients with HFpEF compared to controls. In the skin of patients with HFpEF we also observed a reduced number (85±27 vs 130±60 vessels/mm2; p=0.012) but larger average diameter of lymphatic vessels (42±19 vs 26±9 μm2; p=0.007) compared to control subjects. These changes were paralleled by reduced expression of LYVE1 (p&amp;lt;0.05) and PROX1 (p&amp;lt;0.001), key determinants of lymphatic differentiation and function. Whilst patients with HFpEF had reduced peripheral capillary fluid extravasation compared to controls (microvascular filtration coefficient, leg 33.1±13.3 vs 48.4±15.2, p&amp;lt;0.01; trend for arm 49.9±20.5 vs 66.3±30.1, p=0.09), they had lower lymphatic clearance (isovolumetric pressure: leg 22±4 vs 16±4 mmHg, p&amp;lt;0.005; arm 25±5 vs 17±4 mmHg, p&amp;lt;0.001). Conclusions We provide direct evidence of systemic dysfunction and rarefaction of small blood vessels in patients with HFpEF. Despite a reduced microvascular filtration coefficient, which is in keeping with microvascular rarefaction, the clearance of extravasated fluid in HFpEF is limited by an anatomically and functionally defective lymphatic system. Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): British Heart Foundation Centre of Research Excellence Award

Plasma extracellular vesicles modulate immune cell gene expression following myocardial infarction

Abstract Background Myocardial infarction (MI) induces activation of immune cells and alters their gene expression en route to the injured myocardium but the underlying mechanisms coordinating immune cell programming following MI remain unknown. Plasma extracellular vesicle (EV) numbers are elevated in MI, correlate with the extent of myocardial injury and mobilises immune cells from the splenic reserve to peripheral blood. Here, we describe the role of plasma EV-microRNAs (miRs) in the modulation of peripheral blood mononuclear cell (PBMC) transcriptomes post-MI. Methods PBMCs were exposed to plasma EVs followed by whole transcriptome RNA-sequencing. Plasma EVs were isolated by size-exclusion chromatography and ultra-centrifugation (2 hours at 120,000 x g) from patients presenting with ST-segment elevation MI (STEMI) (N=9) and non-STEMI (NSTEMI) (N=11) control patients. Plasma EVs were characterised by western blot and Nanoview for EV markers CD9 and CD63, transmission electron microscopy (TEM) for morphology and Nanoparticle Tracking Analysis for size and concentration. High sensitive C-reactive protein (hs-CRP) and PCSK9 were determined in plasma by ELISA and compared to plasma EV number using Pearson's correlation. Plasma EV-miRs were measured by Agilent microarray and miR-mRNA putative targets assessed by TargetScanHuman. Results Plasma EVs were positive for EV markers CD9 and CD63, displayed typical EV morphology by TEM and had a heterogeneous size and concentration distribution profile as determined by Nanoparticle Tracking Analysis. Plasma EV number correlated significantly with hs-CRP at presentation (R2= 0.20 and P&amp;lt;0.05). miRNA array analysis revealed STEMI plasma-EVs contained significantly more miR-4487 (P&amp;lt;0.001), miR-6511b-5p (P&amp;lt;0.001), miR-4508 (P&amp;lt;0.001) vs NSTEMI control plasma-EVs at the time of injury. STEMI-plasma-EVs induced differential gene expression in PBMCs vs. NSTEMI-control-plasma-EVs. Gene set enrichment analysis (GSEA) showed STEMI-plasma-EVs upregulated pro-inflammatory pathways including: interferon-α (IFN-α) (P&amp;lt;0.01), IFN-γ (P&amp;lt;0.01), tumour necrosis factor-α (TNF-α) (P&amp;lt;0.01) and interleukin-6 (IL-6)-STAT3 signalling of the acute phase response (P&amp;lt;0.05). miR-4487 (P&amp;lt;0.001) and miR-6511-5p (P&amp;lt;0.05) predicted mRNA targets were significantly enriched in PBMC transcriptomes following treatment with STEMI plasma-EVs. Conclusions Plasma EVs mediating immune cell transcriptional programming following MI by promoting inflammatory pathways in PBMCs is a novel finding. Targeting PBMCs with EVs may allow modulation of the immune response following myocardial injury, to perturb inflammatory immune mediated damage following ischaemic injury. Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): British Heart Foundation Centre of Research Excellence Awards, British Heart Foundation Project Grant, Novo Nordisk Fonden the Tripartite Immunometabolism Consortium and Wellcome Institutional Strategic Support Fund (ISSF)

Determinants of statin initiation and discontinuation in the secondary prevention of atherosclerotic cardiovascular disease in Scotland during 2009–2017

Abstract Background Previous studies have shown that use of statins for secondary prevention of cardiovascular disease (CVD) is suboptimal. However, the role of particular individual characteristics at different treatment stages is limited. Purpose To use large-scale population-wide individual patient data to investigate reasons for suboptimal use of effective CVD medications, in order to identify patient groups that could be targeted to improve medication adherence. Methods This observational longitudinal study used anonymised linked NHS Scotland administrative data (General/Acute Inpatient and Day Case, the National Records of Scotland and the Prescribing Information System) for all individuals hospitalised for an atherosclerotic CVD event (based on ICD-10 discharge codes) in Scotland between 1 April 2009 and 31 December 2017. Statin initiation was defined as individuals being prescribed statin therapy within 90 days from index discharge and dispensed within 60 days from that prescription. Discontinuation was defined as the start of first statin treatment gap of 180 days or more since initiation. Multivariate logistic regression and Cox proportional hazards models were used to study the relevance of patient characteristics (e.g. demographic, clinical, socio-economic) and admission calendar year to the likelihood of, respectively, initiating or discontinuing statin treatment. Findings are reported for all CVD events and, separately, for myocardial infarction (MI), ischaemic stroke (IS) and peripheral arterial disease (PAD). Results Of the 178,113 patients hospitalised for CVD, 19% did not initiate statin treatment. Among the 144,077 patients initiating (40% on high-intensity statins, as defined by NICE guidelines), 25% discontinued treatment within 2 years. Initiation was less common in women (29% less likely than men), older people (22% and 50% less likely for patients in their 70s and 80s respectively vs. 60s), people living in more deprived areas, people receiving specialist mental health care, people with multiple morbidities and people not taking statin prior to admission (Figure). Most of these characteristics were also associated with a decreased likelihood of initiating high-intensity statins, as well as an increased risk of discontinuing statin therapy. In later years, levels of statin initiation, including on higher-intensity statins (58% of statin initiators in 2015–17 vs. 32% in 2009–11), and statin persistence have improved. Conclusions Rates of statin initiation and discontinuation remain suboptimal, especially among women, older people and people with multiple morbidities or mental health illness. Figure 1. Patient characteristics associated with initiation and discontinuation of statin treatment after atherosclerotic CVD event. Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): British Heart Foundation Centre of Research Excellence (Pump Priming Scheme), Medical Research Council UK

Distinct transcriptional and functional features of regenerative mouse neonatal cardiac macrophages

Abstract Background The myocardium of neonatal mice is able to regenerate after myocardial infarction (MI), whereas in adults the formation of scar predominantly occurs following heart injury. Macrophages are involved in the fibrotic response in adult mouse hearts, but also required for successful regeneration in neonates. Recent work has demonstrated that macrophages directly contribute collagen to scar formation following MI. Furthermore, neonatal and adult cardiac macrophages have divergent transcriptional responses to injury. Here, we describe differential transcriptomes and signalling pathways of these functionally distinct neonatal resident cardiac macrophages. Methods Hearts from neonatal P1, P7 and adult CD1 mice (n=3 per group) were digested with collagenase to produce a single cell suspension. Macrophages were isolated by FACS and identified as Ly6G, F4/80+, LyChi/lo cells. Macrophage whole transcriptomes were measured by Illumina RNA-sequencing. Transcript abundance was quantified from raw reads by Salmon and analysis of differentially expressed (DE) genes was carried out with DESeq2. Gene Ontology (GO) enrichment analysis of DE genes was performed with PANTHER. Genes were ranked according to p-value for differential expression, then these ranked genes were used for Gene Set Enrichment Analysis (GSEA) to detect enriched gene sets from the Molecular Signatures Database. Results RNA-sequencing of transcriptomes from neonatal P1, P7 and adult mouse macrophages from hearts highlighted distinct gene expression profiles. The greatest differences were between P1 vs. adult (4,494 differentially expressed (DE) genes at p&amp;lt;0.05) and P7 vs. adult (3,347 DE genes), whereas P1 and P7 macrophages were relatively similar (478 DE genes). A set of 171 genes was found to be DE in P1 vs. P7 and adult macrophages. This P1-specific gene set was highly enriched for GO terms including matrix disassembly (29-fold enrichment, p&amp;lt;0.05) and regulation of chemokine production (12-fold enrichment, p&amp;lt;0.05). GSEA analysis highlighted key functional pathways that were differentially regulated in P1 macrophages, including oxidative phosphorylation and glycolysis, the E2F transcription factors and cell cycle regulators Myc and p53. Conclusions We highlight key genes and pathways distinct to resident neonatal cardiac macrophages to determine the basis for the regenerative capacity of these cells. Interestingly, while genes associated with extracellular matrix were previously shown to be altered after MI in neonatal macrophages, similar differences were also observed here in the basal state of resident neonatal macrophages. We also identified transcriptional and cell cycle regulators linked to the programming and regenerative capacity of these macrophages. The functional differences found in neonatal macrophages might represent potential targets for novel therapeutics. Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): Novo Nordisk Fonden the Tripartite Immunometabolism Consortium (NNF15CC0018486), British Heart Foundation Centre of Research Excellence Awards (RE/13/1/30181)

A short term treatment with a Mek1/2 inhibitor promotes myocardial arteriogenesis and perfusion in vivo: focus on cardiac mural cells

Abstract Background Arteriogenesis is crucial for heart recovery after ischaemia, but cellular and molecular mechanisms able to foster this phenomenon are still poorly characterised. Purpose To discover novel pro-arteriogenic approaches by exploiting cardiac mural cells endowed with arteriogenic capacity: pericytes (PCs) and vascular smooth muscle cells (VSMCs). Methods and results We derived human and murine CD31neg CD34pos cardiac PCs (cPCs) from myocardial samples of adult subjects and confirmed the pericyte phenotype and function in vitro. We discovered that the withdrawal of EGF and bFGF from the culture media induces the differentiation of cPCs into contractile VSMCs. Molecular investigations of pathways associated with the two factors showed that the Mek1/2-Erk1/2 signalling exerts an inhibitory transcriptional control on contractile VSMC genes. Screening of compounds able to interfere with this pathway revealed that PD0325901 – a potent Mek1/2 inhibitor (MeKi) tested in clinical trials for the treatment of cancer – activates the VSMC phenotype in cPCs. We observed a similar effect on coronary artery VSMCs. Next, we interrogated the effect of PD0325901 on cardiac arteriogenesis in vivo. Adult C57BL6/J mice were given the MeKi 10 mg/kg/day or vehicle (DMSO), orally for 14 days (n=11/group). At the endpoint, echocardiographic evaluation of left ventricle (LV) function and dimensions (n=6/group) showed no difference in comparison with the respective baseline in both groups. Effective inhibition of Mek1/2 in the heart of PD-treated mice was confirmed by the reduced immunostaining for the phosphorylated form of Erk1/2. The MeKi cardiotoxicity was ruled out by assessment of cardiomyocytes and vascular cells apoptosis (Tunel) and plasmatic levels of cTn-I. Histological analyses of the hearts (n=5/group) showed an increase in small arterioles (diameter &amp;lt;20μm) density in the LV of PD-mice compared with the DMSO group (16.4 vs 11.7 art/mm2). No change was observed for the capillary density. The drug promoted the maturation of VSMCs within both small and large (&amp;gt;20μm) arterioles, as shown by the higher ratio between the areas of the vascular wall occupied by the mature contractile marker SM myosin heavy chain and the synthetic/early contractile marker alpha-SM actin (αSMA). The PD treatment reduced the fraction of small arterioles covered with a CD34pos layer (53% vs 70% of total arterioles), along with a lower ratio between the areas occupied by adventitial CD34pos cells and αSMApos VSMCs, suggesting a contribution of cPCs to the arteriolar remodelling. Last, the drug improved the LV myocardial perfusion in the PD- vs the DMSO-group (6.8 vs 5.3 ml/min/g of LV tissue, n=6/group). Conclusions We show that a short treatment with a Mek1/2 inhibitor stimulates myocardial arteriogenesis and perfusion without either inducing cardiotoxicity or deteriorating heart function. This may be a novel, intriguing approach to promote therapeutic arteriogenesis. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): British Heart Foundation Centre for Vascular Regeneration II