bluuebunny/crossref_metadata_2025 huggingface dataset:
Prevalence, incidence, and outcomes across cardiovascular diseases in homeless individuals- a case-control study using national linked electronic health records
Authors: Banerjee A , Nanjo A , Evans H , Direk K , Hayward A , Story A
Published: November 2020

Background

Homelessness is increasing globally. The risk and burden of cardiovascular disease (CVD) are higher in homeless than in housed individuals but the population-based analyses, required for planning of effective interventions and policies, are lacking.

Purpose

Using national electronic health records (EHRs), we investigated prevalence, incidence and outcomes across a range of CVDs among homeless individuals.

Methods

Using linked UK primary care EHR and validated phenotypes, we identified homeless individuals aged ≥16 years between 1998 and 2019, and age- and sex-matched housed controls in a 1:5 ratio. For twelve CVDs (stable angina; unstable angina; myocardial infarction; sudden cardiac death or cardiac arrest; unheralded coronary death; heart failure; transient ischaemic attack; ischaemic stroke or stroke not further specified; subarachnoid haemorrhage; intracerebral haemorrhage; peripheral arterial disease; abdominal aortic aneurysm), we estimated prevalence, incidence and 1-year mortality risks, comparing homeless and housed groups.

Results

We identified 8492 homeless individuals and 32134 matched controls. Comorbidities and risk factors were more prevalent in the homeless group, e.g. smoking: 78.1% vs 48.3% and atrial fibrillation: 9.9% vs 8.6%, p<0.001. CVD prevalence (11.6% and 6.5%), incidence (e.g. incidence rate ratio, 3.36, 1.99–5.66 and 2.18, 1.50–3.16 in men and women, respectively, aged <35 years), and 1-year mortality risk were higher (adjusted hazard ratio 2.30, 1.70–3.12), and onset was earlier (difference: 4.6, 2.8–6.3 years, p<0.001), in homeless, compared with housed people. Other than cerebrovascular and peripheral vascular diseases in women (where numbers of events were small), homeless individuals had higher incidence of CVD in all three arterial territories than housed people.

Conclusions

CVD in homeless individuals has high prevalence, incidence and 1-year mortality risk with early onset, and high burden of risk factors. Health and social care strategies in inclusion health should reflect this high burden of treatable risk factors and disease.

Incidence of CVD in homeless individuals

Funding Acknowledgement

Type of funding source: Public grant(s) – National budget only. Main funding source(s): National Institute of Health Research
Adapting neural network models to predict 10-year CVD development based on regional data calibration
Authors: Korsakov I , Gavrilov D , Serova L , Gusev A , Novitskiy R , Kuznetsova T
Published: November 2020

Background

The used tools for prediction the individual risk of developing cardiovascular diseases and their complications using machine learning methods have proven better prognostic value in comparison with commonly used scales (e.g., Framingham, SCORE). To create such methods, the long-term accumulation of large amount of qualitative data are required. Moreover, to improve the accuracy of models, it is necessary to take into account regional characteristics that affect health: ethnic, nutritional characteristics, climatic conditions, living standards and medical care. These regional characteristics could significantly affect the development and outcomes of CVDs. However, the amount of regional data is not enough to build a qualitative model. Therefore, it is proposed to create models based on publicly available data and validate them on regional medical data sufficient for validation and calibration.

Methods

Two models were trained using data from the Framingham study. Model 1 was trained on 2 588 patient data and predicts a 10-year CVD probability according to the following risk factors: age, gender, cholesterol, HDL, smoking, SBP, and BP medications. Model 2 was trained on 4,363 patient data and predicts a 10-year death probability from CVD according to the following criteria: age, gender, cholesterol, smoking, SBP, BMI, heart rate. To retrain the obtained models, we used dataset created from data from patients in the northwestern part of Russia. The dataset consists of 438 patients, including the signs used in the trained models. This dataset includes CVD and death from it during a 10-year follow-up

Evaluation

We used randomized data splitting: divided the dataset into a training and a test set with an 80/20 proportion. The models was implement with keras convolution neural network (CNN) using 3 hidden layers. For data validation was used a 10 K-fold method.

Results

We compared the initial model metrics and those obtained after local data retraining. The accuracy of model 1 before retraining is 78%, after – 81.3%, the area under the ROC curve (AUC) before retraining: 0.77 (at 95% CI: 0.72–0.82C), after – 0.803. The accuracy of model 2 before retraining is 79%, after – 85.6%, the area under the ROC-curve (AUC) before retraining: 0.78 (at 95% CI: 0.72–0.82), after – 0.828.

Conclusion

Using this method of retraining predictive models, we can take into account local characteristics of the population and significantly increase the accuracy of predicting events. Expand the population to use the model according to local characteristics.

Funding Acknowledgement

Type of funding source: Private company. Main funding source(s): OOO K-SkAI
National cardiac registries: a systematic review
Authors: Dawson L , Biswas S , Stub D , Lefkovits J , Burchill L , Reid C , Eccleston D
Published: November 2020

Background

Over the last thirty years, there has been exponential growth in the number and scale of national cardiovascular disease registries. We aimed to provide a comprehensive outline of contemporary national cardiac registries across all subspecialties.

Methods

We performed a systematic literature review by searching OvidMedline in August 2019 to identify registries relating to six pre-specified domains (Table). For inclusion, registries had to be national in nature, actively collecting data, and publishing either peer-reviewed publications or online reports.

Results

A total of 24,076 records were identified from six domain-specific Medline searches; 24,057 abstracts were screened with 19,435 non-relevant records excluded; 4,624 full texts were screened with 4,473 non-relevant texts excluded; and 151 registries met inclusion criteria representing 51 countries. Of these, 15 related to cardiac surgery, 27 to arrhythmia (17 device, 5 ablation, 7 atrial fibrillation), 21 to congenital heart disease (14 general, 2 interventional, 4 surgical, and 1 disease specific), 43 to coronary disease or percutaneous coronary intervention (22 PCI, 21 CAD), 27 to heart failure (13 heart failure, 5 transplant, 2 mechanical support, 7 disease specific), and 18 related to structural intervention (3 any, 13 transcatheter aortic valve replacement, 2 mitral intervention). Nine national registries (USA, Sweden, Finland, Denmark, UK, Portugal, Norway, Taiwan, and Singapore) covered multiple domains. Quality scoring using the Monash University Clinical Quality Registry Grading System (a composite score of recruitment, and data completeness, definitions, reliability and validation), demonstrated marked heterogeneity in quality between registries.

Conclusions

Cardiac registries have seen rapid growth, however the use and quality among various subspecialties differs markedly across world regions. Given the multiple benefits, clinicians, funders and health bureaucrats should be encouraged to focus on the range, quality and uptake of national registries.

Figure 1

Funding Acknowledgement

Type of funding source: None
Mutations in CHMP4C cause dilated cardiomyopathy via dysregulation of autophagy
Authors: Zhou N.W , Tang L , Jiang Y.Y , Li X.J , Zhao W.P , Pan C.Z , Wang X.L , Shu X , Qiu Z.L , Ge J.B
Published: November 2020

Background

Gene mutations have been implicated in DCM. However, due to the difficulty of clinical genetic diagnosis, more causal genes potentially related to DCM remain to be discovered.

Methods

We screened for gene mutations in more than 400 cases from families with hereditary cardiovascular disease using whole-exome sequencing. Then we validated biological functions of CHMP4C mutations in zebrafish models. To further assess the mechanism of CHMP4C mutations, we evaluated the potential signaling pathway in the cells.

Results

We identification of CHMP4C variants that segregated with DCM variants in four families from a total of 411 families via whole-exome sequencing. We further validate the function of CHMP4C in heart function in zebrafish models and found that over-expression of CHMP4C variants in zebrafish resulted in cardiac malformation, pericardial edema and increased heart rate, consistent with CHMP4C mutation-associated findings in DCM patients. Furthermore, we found that mutations in CHMP4C impaired autophagy and activated apoptosis in HEK293T cells, suggesting that the molecular mechanism of CHMP4C is involved in heart development.

Conclusions

CHMP4C is a novel candidate gene for DCM and may play a critical role in cardiac development by regulating autophagy.

Funding Acknowledgement

Type of funding source: None
Epicardial adipose tissue: the genetics behind an emerging cardiovascular marker
Authors: Sousa J.A , Serrao M.G , Temtem M , Pereira A , Santos M , Mendonca F.V , Monteiro J.P , Ferreira A , Freitas P , Henriques E , Ornelas I , Freitas A.I , Freitas A.D , Reis P , Mendonca M.I
Published: November 2020

Background

Increasing evidence points epicardial adipose tissue (EAT) as an emerging cardiovascular risk marker. Whether genetic polymorphisms are associated with a higher EAT burden is still unknow. Genetic risk score (GRS) is an emerging method that attempts to establish correlation between single nucleotide polymorphisms (SNPs) and clinical phenotypes.

Aim

Evaluate the role of genetic burden and its association to EAT.

Methods

996 patients (mean age 59±8, 78% male) were prospectively enrolled in a single center. EAT was measured on cardiac CT using a modified simplified method. Patients were divided into 2 groups (above vs. below the median EAT volume).

We studied different polymorphisms across the following gene-regulated pathways: oxidation, renin-angiotensin system, cellular, diabetes/obesity and dyslipidemia pathways. Genotyping was performed by TaqMan allelic discrimination assay. A multiplicative genetic risk score (mGRS) was constructed and represents the genetic burden of the different polymorphisms studied. To evaluate the relation between genetics and EAT volume, we compared both groups by: global mGRS, gene cluster/axis mGRS and individual SNPs.

Results

Patients with above-median EAT volume were older, had higher body mass index (BMI) and higher prevalence of hypertension, diabetes and dyslipidemia (p<0.05). Patients with higher EAT volumes presented a higher global mean GRS (p<0.001), with the latter remaining an independent predictor for higher EAT volumes (OR 1.3, 95% CI 1.2–1.5), alongside age and BMI.

In the analysis by gene clusters, patients with more epicardial fat consistently presented a higher polymorphism burden (translated by a higher mGRS level) across numerous pathways: oxidation, renin-angiotensin system, cellular, diabetes/obesity and dyslipidemia. After adjusting for confounders and other univariate predictors of higher fat volume, the following have emerged as independently related to higher EAT volumes: mGRS comprising the genes of different clusters, age and BMI.

Amongst the 33 genes analyzed, only MTHFR677 polymorphisms (a gene with a critical role in regulating plasma homocysteine levels) emerged as significantly related to higher EAT volumes in our population (OR 1.4, 95% CI: 1.100–1.684, p=0.005).

Conclusion

Patients with a higher polymorphism burden in genes involved in the oxidation, renin-angiotensin, cellular, diabetes/obesity and dyslipidemia pathways present higher levels of epicardial fat. This potential association seems to be independent from the expected association between epicardial fat and cardiovascular risk factors. To our knowledge, this is the first time such genetic profiling has been done, casting further insight into this complex matter.

Funding Acknowledgement

Type of funding source: None
Aetiology-dependent impairment of mitochondrial function in the failing human heart
Authors: Borger J , Scheiber D , Horn P , Pesta D , Boeken U , Akhyari P , Lichtenberg A , Kelm M , Roden M , Szendroedi J , Westenfeld R
Published: November 2020

Background

Alterations of mitochondrial function have been identified to play a role in Heart Failure (HF) pathophysiology. Oxidative phosphorylation (OXPHOS) capacity of the myocardium was shown to be reduced in the failing heart. Ineffective mitochondrial function promotes formation of reactive oxygen species (ROS) that may affect remodelling in ischemia. Thus far, human mitochondrial function comparing dilated cardiomyopathy (DCM) and ischemic cardiomyopathy (ICM) resembling the main aetiologies of heart failure with reduced ejection fraction (HFrEF) has not been investigated.

Purpose

We hypothesised that

1. ROS production is elevated in left ventricular myocardial tissue specimens of ICM patients compared to DCM.

2. Mitochondrial OXPHOS capacity is higher in left ventricular myocardial tissue specimens of DCM compared to ICM patients.

Methods

Myocardial tissue was obtained from the left ventricular apex from 63 patients (38 ICM, 25 DCM) with advanced HFrEF requiring implantation of a Left Ventricular Assist Device (LVAD). We performed high-resolution respirometry (HRR, OROBOROS Oxygraph-2k) in saponine-permeabilised myocardial fibres and measured ROS production fluoroscopically via the Amplex Red method. Statistical analysis was conducted using GraphPad Prism 7 and IBM SPSS v26.0.

Results

Groups were of comparable age (61.5±1.2 vs. 59.3±2.4 years, p=n.s.), sex (87% vs 85% male, p=n.s.), diabetic status (32% vs 38.4% type 2 diabetes mellitus, p=n.s.), and body mass index (28.1±0.8 vs. 26.3±1.1 kg/m2, p=n.s.). We detected reduced myocardial mitochondrial OXPHOS capacity in ICM under state 3 conditions by about 15% (68.7±34.0 vs. 80.9±30.5 pmol/(s*mg), p<0.05), after addition of Glutamate by 25% (78.9±38.7 vs. 104.8±41.2 pmol/(s*mg), p<0.01) as well as after Succinate (115.5±65.5 vs. 155±62.0 pmol/(s*mg), p<0.01), uncoupling agent FCCP (114.1±56.8 vs. 150.5±47.3 pmol/(s*mg), p<0.01), and by about 40% after addition of Complex I inhibitor Rotenone (55.5±25.9 vs. 96.9±28.0 pmol/(s*mg), p<0.001). We detected no difference in ROS production between ICM and DCM (0.6±0.05 vs. 0.76±0.08 pmol/(s*ml), p=n.s.).

Conclusion

This is the first human study deciphering distinct alterations in mitochondrial function (OXPHOS capacity) in ventricular myocardium of HFrEF patients. Future studies may address how distinct metabolic patterns at the time of implantation may relate to long-term outcome of HFrEF in terms of remodelling and recovery.

Funding Acknowledgement

Type of funding source: Public grant(s) – National budget only. Main funding source(s): DFG (German Research Foundation)
RBP4 induces pyroptosis in cardiomyocytes via activating NLRP3/Caspase-1/GSDMD pathway in acute myocardial infarction
Authors: Gao W , Zhang K.Z
Published: November 2020

Aims

Myocardial infarction (MI) is the most common cause of cardiac morbidity and mortality worldwide. Pyroptosis is a novel form of inflammatory cell death that could be driven by the nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome activation following MI. Retinol binding protein 4 (RBP4) is an pro-inflammatory adipokine with adverse effects on cardiovascular system. We investigated the role and underlying mechanism of RBP4 in inducing pyroptosis in cardiomyocytes in acute MI.

Methods and results

We established a mice MI model which ligated with the left anterior descending coronary artery and a cardiomyocytes injury model treated by oxygen-glucose deprivation (OGD). Western blot showed that RBP4 expression was exclusively increased in the heart following MI, as RBP4 expression remains unchanged in adipose tissues and liver. Cardicac pyroptosis was induced in MI mice as evidenced by increased expression of pyroptosis-related proteins, including NLRP3, Caspase-1, gasdermin D (GSDMD), IL-1β and IL-18. Moreover, increased cardiac RBP4 level was positively correlated with serum IL-1β and IL-18 levels in MI mice. Immunofluorescence identified that RBP4 was expressed in cardiomyocytes and was up-regulated along with increased pyroptosis-related proteins after treatment with OGD. Transfection of RBP4 adenovirus promoted pyroptosis while inhibition of RBP4 ameliorated OGD-induced pyroptosis in cardiomyocytes. Mechanically, transfection of NLRP3 siRNA or GSDMD siRNA along with RBP4 adenovirus could attenuate RBP4-induced pyroptosis in cardiomyocytes. Furthermore, in situ injection of adenovirus containing RBP4 siRNA could also attenuate pyroptosis and cardiac inflammation following MI in mice.

Conclusion

RBP4 is presented in cardiomyocytes and induces cardiac pyroptosis via activating NLRP3/Caspase-1/GSDMD pathway in acute MI

Funding Acknowledgement

Type of funding source: Foundation. Main funding source(s): National Nature Science Foundation of China
Procalcific effect of proprotein convertase subtilisin/kexin type 9 (PCSK9) in chronic kidney disease (CKD)
Authors: Ferri N , Lupo M.G.P , Rattazzi M.R
Published: November 2020

Aim

PCSK9 has been recently associated with a higher rate of calcification in hypercholesterolemic, diabetes and CKD patients. The aim of this study was thus to investigate the role of PCSK9 in VC process, under uremic condition, both on in vivo and in vitro experimental settings.

Methods

Sprauge Dawley rats were fed a standard diet (SD, n=11) or uremic diet (UD, n=11) for 6 weeks. Calcium crystals in aortas were visualized by von Kossa staining and quantified by a colorimetric assay and plasma total cholesterol determined. Control and PCSK9-overexpressing smooth muscle cells (SMCsPCSK9) were cultured with 2.5% FCS ± Pi for 7 days. Hydroxyapatite deposition by SMCs was measured by a colorimetric assay. The number and the content of pro-calcific extracellular vesicles (EVs) budding from the cells were determined.

Results

The hyperphosphatemia secondary to CKD lead to rat aortic calcification (+7.3-fold) and a significant increase in TC and PCSK9 levels (+1.4 and +2.7-fold, respectively). Higher expression of PCSK9 was also observed in kidney (+4.8-fold) and liver (+1.5-fold). SMCsPCSK9 showed higher extracellular calcium deposition (+1.4-fold) in response to Pi and increase EVs production (+7-fold). The incubation of control cells with recombinant PCSK9 did not induce extracellular calcium deposition.

Conclusions

Our study suggest a positive effect of intracellular PCSK9 on vascular calcification in CKD condition. Pro-calcific budding of EVs seems one of the possible mediators of this process.

PCSK9 and calcification

Funding Acknowledgement

Type of funding source: None
Evaluation of noninvasive electrophysiological imaging accuracy for focal atrial arrhythmias
Authors: Budanova M , Chmelevsky M , Zubarev S , Treshkur T , Lebedev D
Published: November 2020

Background

High accuracy of noninvasive electrocardiographic imaging (ECGI) has recently been shown for topical diagnostics of ventricular arrhythmias. However, the precision of diagnostics of atrial focal arrhythmias requires clarification. To estimate the accuracy of ECGI for premature atrial contraction (PAC) we performed atrial pacing in patients with CRT system and compared early activation zone (EAZ) with pacemaker's tip location.

Purpose

To determine the accuracy of ECGI for focal atrial arrhythmias using atrial pacing.

Methods

Twenty-six patients (m/f – 18/9), age (min–max) 52 (26–78) with CRT system and pacemaker's tip location in the right atrium (RA) appendage underwent ECGI (“Amycard 01C”) in combination with CT or MR imaging. Thirty-four atrial pacing (mono- and bipolar) was performed in all patients using standard amplitude 1.5–3.8 mV. Epi-/endocardial polygonal heart models were created and isopotential maps were calculated. The distance between EAZ and the pacemaker's tip were measured for ECG recordings without using the isoline filter on endocardial surface (Fig. 1) as well as for epicardial surface. The time between epicardial and endocardial EAZ breakthrough was calculated also.

Results

On endocardial surface the EAZ was located in RA appendage, the base of superior cava vena or superior lateral RA wall. The distance (mm) (Me (min; max)) between EAZ and the pacemacer's tip was 28 (6; 68). For epicardial surface in most cases the EAZ was also located in RA appendage, the base of superior cava vena or superior lateral RA wall. In two cases the EAZ was located in inferior septal RA wall, in one case - in superior septal RA wall and in five cases the EAZ was undetectable. The distance between EAZ and the pacemacer's tip was 22 (6; 48). The time (ms) (Mean; Me (min; max)) between EAZ of the endocardial and epicardial surfaces was 16; 7 (0; 68).

Conclusion

ECGI allows to assess the location of focal atrial arrhythmias on endocardial surface and sometimes on epicardial surface also within the three segments. The results of this study revealed that accuracy of ECGI for atrial arrhythmias is worse than for ventricular arrhythmias. However, it is better on epicardial surface of atrium when EAZ can be determined.

Funding Acknowledgement

Type of funding source: None
Acute effects of a mavacamten-like myosin-inhibitor (MYK-581 in a feline model of obstructed hypertrophic cardiomyopathy: evidence of improved ventricular filling (beyond obstruction reprieve)
Authors: Ferguson B.S , Stern J.A , Oldach M.S , Ueda Y , Ontiveros E.S , Napierski N.C , Del Rio C.L , Harris S.P
Published: November 2020

Introduction

Hypertrophic cardiomyopathy (HCM) is a progressive disease characterized by cardiac remodeling, hyperdynamic contraction, and impaired ventricular filling that can lead to dynamic left-ventricular outflow-track (LVOT) obstruction and exertional intolerance. Direct myosin-inhibition with mavacamten can normalize contractility and improve exercise capacity in patients with oHCM, providing sustained symptomatic relief. However, mavacamten can also improve ventricular filling by limiting residual cross-bridges during diastole, and therefore, may offer cardiac benefits beyond obstruction reprieve. This study leveraged a feline model of oHCM, cats with the A31P MYBPC3 variant, to study the acute in vivo effects of MYK-581, a mavacamten surrogate, on cardiac hemodynamics and filling.

Methods

A31P-homozygous cats with HCM (A31P, n=10) and wild-type healthy controls (CTRL, n=9) were anesthetized and instrumented for invasive pressure-volume (PV) measurements as well as trans-thoracic echocardiographic recording. A subset of cats were assigned to receive either vehicle (VEH, n=7) or MYK-581 (MYK, n=8) with a short IV infusion. Cardiac hemodynamics, function, and geometry were assessed at steady state before and during dobutamine challenges (2.5 μg/kg/min IV).

Results

A31P cats had thicker ventricular walls (6.4±0.1 vs. 5.2±0.2 mm, P<0.05) and hyperdynamic contraction (FS: 61±4 vs. 50±3%, P<0.05) relative to controls and presented with dynamic LVOT obstruction in 54% of cases. HCM cats had elevated end-diastolic pressures (17±1.4 vs. 9±1.0 mmHg, P<0.05), with prolonged time constants of relaxation (60±4.1 vs. 36±2.4 ms, P<0.05) and elevated end-diastolic stiffness (Eed: 0.44±0.06 vs. 0.25±0.01 mmHg/mL). Acute treatment with MYK-581 alleviated LVOT obstruction (0% vs. 38%), normalized contractility (FS: −7±2%), and increased systolic/diastolic chamber dimensions (e.g., LVIDd: +13±4%) (all P<0.05), while reducing EDP (15±2 to 13±2 mmHg, P<0.05), suggesting acute improvement in ventricular distensibility. Indeed, MYK-581 treatment reduced end-diastolic stiffness (Eed: 0.48±0.11 vs. 0.36±0. 10 mmHg/mL, P<0.05) and normalized trans-mitral motion patterns during filling.

Conclusions

Bred cats, homozygous for the A31P MYBPC3 variant, presented a cardiac phenotype that models multiple characteristics of the human oHCM phenotype including dynamic LVOT obstruction. Acute treatment with the mavacamten surrogate, MYK-581, not only alleviated hypercontractility and LVOT obstruction, but improved ventricular filling and end-diastolic pressures. Taken together, these pre-clinical observations show potential salutary effects beyond obstruction relief in patients with HCM.

Funding Acknowledgement

Type of funding source: Private company. Main funding source(s): MyoKardia
Reciprocal modulation of Linc-223 and its ligand miR-125a on the basis of platelet function level
Authors: De Rosa S , La Bella S , Canino G , Siller-Matula J , Eyleten C , Postula M , Tamme L , Iaconetti C , Sabatino J , Polimeni A , Sorrentino S , Gareri C , Proto L , Strangio A , Indolfi C
Published: November 2020

Background

Cardiovacular diseases (CVD) are the leading cause of death worldwide. Platelet play a key role in the pathophysiology of multiple CVD. LincRNAs are long non-coding RNAs transcribed from intergenic DNA segments. Some members of this class were recently associated with human disease. Linc-223 is co-transcribed together with mir-223 and was recently shown to bind to miR-125. Both miR-223 and miR-125 are highly expressed in platelets.

Purpose

In light of the capability of Linc-223 to bind to miR-125, we aimed to investigate whether their reciprocal expression levels might reflect the degree of platelet activity.

Methods

RNA was extracted using miRVANA. MiRNAs and lncRNAs were measured by means of quantitative Real Time RT-PCR.

Results

We found a significant reduction of Linc-223 levels (p<0.05) along with a significant increase in miR-125 levels (p<0.05) after initiation of any antiplatelet treatment (n=30) compared to naïve patients (n=10). Moreover, the upgrade to a higher-intensity antiplatelet treatment with ASA+ticagrelor from ASA+clopidogrel (n=30) was associated to a further down-regulation of Linc-223 (p<0.05) along with a further increase of miR-125. (p<0.05). Finally, these results were validated in a larger cohort of 300 patients from the ATLANTIS study, demonstrating significant modulation of both miR-223 and miR-125 in patients with high on-treatment platelet aggregation levels compared to antiplatelet-responsive patients.

Conclusions

We identify a reciprocal modulation of Linc-223 and miR-125, its ligand upon different levels of platelet aggregations. These results are compatible with previous evidence from patients with Acute Myeloid Leukemia, that Linc-223 might bind to and sponge miR-125, inhibiting its effect. These results suggest that plasma levels of Linc-223, miR-125 and miR-223 might be used a biomarkers of platelet finction in a clinical context, for risk stratification of patients or to assess the responsiveness to antiplatelet treatments.

Funding Acknowledgement

Type of funding source: None
The predictive role of endothelial glycocalyx in primary prevention of cardiovascular events:a 6 year follow-up study
Authors: Ikonomidis I , Thymis J , Katsanos S , Kousathana F , Varoudi M , Triantafyllou C , Kostelli G , Triantafyllidi H , Lambadiari V , Papadavid E , Plotas P , Tsilivarakis D , Benas D , Katogiannis K , Iliodromitis E.K
Published: November 2020

Introduction

Endothelial glycocalyx is a layer of glycoproteins on endothelial cells preventing vascular wall permeability to circulating proteins and blood cells. Perfused Boundary Region (PBR) of arterial microvessel is a non-invasive marker of endothelial glycocalyx thickness, as increased PBR indicates a reduced endothelial glycocalyx thickness. Cardiovasular risk factors and cardiovascular diseases have been associated with impaired PBR. However, PBR has not been studied as a predictor of major adverse cardiovascular events (MACE) in primary prevention.

Methods

We studied 400 adults, without history cardiovascular disease. We measured PBR of sublingual vessels with diameter 5–25μm by GlycoCheck Software using a dedicated camera (Sidefield Darkview Imaging, Microscan MV, Nederlands). We recorded a full medical history for each subject. The participants were followed up for a period of 6 years for major cardiovascular events (including Acute Coronary Syndrome, Stroke, decompensated acute heart failure, cardiovascular death and death of all causes).

Results

The mean age of participants was 49±10 years, 52% were males, 65% active smokers, 33.3% had hyperlipidemia, 12.8% type 2 Diabetes respectively, 30% hypertension and 20.5% had positive family history of premature coronary artery disease. The median PBR value was equal to 1.15 μm. After 6 years of follow up, 42 MACE were recorded (7 cardiovascular deaths). In univariate analysis, subjects with PBR values >1.15μm had greater risk of adverse events compared with those that had lower PBR (relative risk RR=2.83, p<0.05). Diabetes and hypertension were also univariate predictors of MACE (RR=3.71 and RR=2.51, p<0.05). In multivariate analysis after adjusting for atherosclerotic risk factors including diabetes and hypertension, PBR remained a an independent predictor of MACE (RR=2.67, p<0.05)

Conclusion

Endothelial glycocalyx thickness appears to be an important independent predictor of adverse cardiovascular outcomes.

Predictive value endothelial glycocalyx

Funding Acknowledgement

Type of funding source: None
CD40 levels in plasma are associated with cardiovascular disease and in carotid plaques with a vulnerable plaque phenotype and remodelling
Authors: Shami A , Edsfeldt A , Shore A.C , Natali A , Khan F , Nilsson J , Lutgens E , Goncalves I
Published: November 2020

Background and purpose

CD40 and CD40 ligand (CD40L) are costimulatory molecules and members of the TNF receptor superfamily well known for their involvement in inflammatory and autoimmune diseases. This study uses two large human cohorts – the SUrrogate markers for Micro- and Macro-vascular hard endpoints for Innovative diabetes Tools (SUMMIT) and the Carotid Plaque Imaging Project (CPIP) – to explore the potential of plasma or intra-plaque expression of CD40 and CD40L as biomarkers and to locally affect plaque stability.

Methods

Proximity Extension Assay (PEA) technique was used to measure soluble CD40 and CD40L (sCD40 and sCD40L) in plasma from 1437 subjects from the SUMMIT cohort, the majority of which (80%) with pre-existing cardiovascular disease, and in atherosclerotic plaque homogenates from 199 subjects of the CPIP cohort undergoing carotid endarterectomy. The Mann-Whitney U test was used to compare groups and Spearman's rank correlation/the Chi-square test was used to assess correlations. Multiple comparisons were corrected for using the Holm-Šídák test. A logistic regression model was used to test for associations with future cardiovascular events and mortality.

Results

In the SUMMIT cohort both plasma CD40 and CD40L levels were elevated in individuals with a history of stroke (p=0.000030 and p=0.020, respectively), while sCD40 levels also were higher in individuals with a prior acute myocardial infarction (p=0.016). Plasma levels of sCD40 correlated with carotid plaque burden (as measured by ultrasound imaging, r=0.355, p<1x10–16) and were associated with future cardiovascular events over a three year-follow up period (p=0.02, hazard ratio 1.3, 95% C.I: 1.042–1.625).

sCD40 and sCD40L were associated with a plaque phenotype characterized by the strong presence of features both of vulnerability such as high content oxidized low-density lipoprotein (LDL; r=0.236, p=0.004 and r=0.259, p=0.0037, respectively) and pro-inflammatory cytokines (e.g. tumour necrosis factor-α: p=3.1x10–7 and p=0.0006, respectively) and low calcium content (r=−0.208, p=0.012 and r=0.268, p=0.00034, respectively).

Conclusion

High plasma CD40 and CD40L levels are associated with symptomatic cardiovascular disease. Plasma CD40 levels correlate with the severity of carotid atherosclerosis and are associated with an increased risk for future cardiovascular events. Additionally, intra-plaque levels are associated with a vulnerable plaque phenotype. Our findings thus support the value of sCD40 and sCD40L both as biomarkers and therapeutic targets for cardiovascular disease.

Funding Acknowledgement

Type of funding source: Foundation. Main funding source(s): The Swedish Heart and Lung Foundation (1) and the Swedish Research Council (2)
Pemafibrate protects the rupture of experimental aortic aneurysm in mice through anti-oxidative stress with induced catalase
Authors: Amioka N , Miyoshi T , Akagi S , Yoshida M , Nakamura K , Morita H , Ito H
Published: November 2020

Background

Abdominal aortic aneurysm (AAA) is a life-threatening disease, while effective and preventive medical treatments remain unestablished.

Purpose

The aim of this study was to evaluate the effect of selective peroxi- some proliferator-activated receptor alpha (PPARα) modulator pemafibrate on AAA formation in mice.

Methods

AAA was induced by subcutaneous infusion of angiotensin II (AngII) in apolipoprotein E-deficient mice for 4 weeks. Treatment by pemafibrate or vehicle was started one week before AngII infusion. Oxidative stress was evaluated by dihydroethidium (DHE) staining.

Results

Prognosis after AngII infusion in pemafibrate-treated mice was significantly better than that in vehicle-treated mice (log-rang test, p=0.035) by reducing the occurrence of fatal AAA rupture. Meanwhile pemafibrate did not significantly reduce maximal diameter of the aorta. Histological findings demonstrated that the expression of collagen in adventitia in pemafibrate-treated mice was significantly greater than that in vehicle-treated mice (p<0.05). Oxidative stress in aorta of pemafibrate-treated mice was significantly reduced comparing to vehicle-treated mice, accompanying by the reduction of mRNA expression of matrix metalloproteinase 2, tumor necrosis factor-α, and interleukin-6 (all p<0.05). Catalase expression in abdominal aortic tissue was increased 1.5-fold in mice treated with pemafibrate than in mice treated with vehicle (p=0.032). In human vascular smooth muscle cells (hVSMC), pemafibrate attenuated AngII-induced oxidative stress (p<0.001), which was canceled by administrating small interfering RNA (siRNA) of PPARα (p<0.001). Furthermore, in hVSMC, pemafibrate increased catalase expression significantly (p<0.001), while this increase was significantly suppressed by knockdown of PPAR-α with siRNA. (p<0.001).

Conclusion

Pemafibrate reduced the rupture of AAA in this murine model, which is associated with anti-oxidative stress via catalase induction.

Funding Acknowledgement

Type of funding source: None
Comparative analysis of various metrics of arterial compliance in the study of chronic inflammatory diseases
Authors: Kerkhof P.L , Djorai R.B , Maslyanskiy A , Kolesova E , Konradi A.O , Shlyakhto E.V , Li J.K.-J
Published: November 2020

Introduction

Arterial compliance (AC), known to depend on age and sex, may be severely compromised in chronic inflammatory diseases. Apart from the various definitions for arterial stiffness that are in vogue, their constituent components are often not unique.

Purpose

This study compares vascular stiffness measures in various inflammatory diseases and explores a more comprehensive description of vessel wall properties by also considering inherent associated companion metrics.

Methods

We retrospectively analyzed arterial pressure, ventricular volume by echocardiography (Vivid-7 ultrasound system), along with carotid–femoral pulse wave velocity (PWV, by SphygmoCor, Atcor). Pulse pressure (PP) equals systolic arterial pressure (SAP) minus diastolic arterial pressure (DAP). Effective arterial elastance (Ea) = 0.9 × SAP/SV where SV is stroke volume. AC=SV/PP (Figure 1A). The intrinsic companion (C) to any established difference-based metric is calculated on the basis of the squared mean, e.g., PPC2 = (SAP2 + DAP2). Clearly, AC depends on two differences, each having a companion.

Results

A total of 177 patients (age range 18 to 80 years, 49 males), characterized by inflammatory disease (51 scleroderma, 62 atherosclerosis, 35 ankylosing spondylitis, and 29 rheumatoid arthritis), yielded reduced levels of vacular compliance irrespective of the method selected. Regression analysis showed weak correlations between the various approaches (i.e., Ea, AC, PWV). Average values for AC are comparable for all 4 diagnostic groups. Ea (P<0.03) and PWV (P<0.014) are lower for ankylosing spondylitis compared to all other groups, likely due to younger age and the higher prevalence of men. Ea is highest in atherosclerosis (P<0.026) versus all other groups. SV is significantly higher in atherosclerosis (P<0.03) compared to scleroderma and rheumatoid arthritis. However, the companions may differ (Figure 1B): PPC is significantly (P=0.017) higher in atherosclerosis (156.3±16.1 mmHg) vs scleroderma (147.2±23.0 mmHg), while SVC in ankylosing spondylitis is higher (P<0.013) than in all other groups.

Conclusions

Arterial stiffness measures show poor correlation, suggesting limitations to their utility when studying the diagnostic groups described here. Consideration of the companion associated with each difference-based metric is warranted in order to perform a comprehensive analysis of clinical data when evaluating the impact on risk factors and prognosis.

Figure 1. A: Compliance components. B: Companions

Funding Acknowledgement

Type of funding source: Public grant(s) – National budget only. Main funding source(s): Russian Science Foundation
Cavin-1 regulates BMP/Smad signaling through the interaction of Caveolin-1 with BMPRII in pulmonary artery endothelial cells
Authors: Tomita S , Nakanishi N , Ogata T , Tsuji Y , Sakamoto A , Higuchi Y , Matoba S
Published: November 2020

Background

Pulmonary hypertension (PH) is a progressive disease associated with poor outcome. Caveolin-1 (Cav1) is a component of caveolae and classified as a related gene of pulmonary arterial hypertension (PAH). Gene mutations of bone morphogenetic protein type II receptor (BMPRII) is a most common cause of PAH. BMPRII is localized in caveolae and associates with Cav1. However, the role of the Caveolin-Cavin system on the BMP/Smad signaling and the PAH progression has not been well-known.

Purpose

The aim of our study is to investigate the relationship between Caveolin-Cavin system and BMP/Smad signaling pathway and explore the mechanism of downstream signal transduction of BMP signaling by the interaction between Caveolin and BMPRII.

Methods

Cav1 knockout mice were used to assess PH and caveolae in pulmonary artery endothelial cells were observed by electron microscope. Cav1 and Cavin-1, which is a component of caveolae and form a complex with Cav1, were knocked-down in human pulmonary artery endothelial cell (hPAEC) using siRNA and phosphorylation of Smad signal was evaluated. Apoptosis of these cells was explored by flow cytometry. We investigated the interaction between Cav1 and BMPRII, and evaluated whether Cavin-1 affects this interaction and signal transduction of BMP signaling.

Results

As previously described, deletion of Cav1 revealed disappearance of caveolae in pulmonary artery endothelial cells (PAECs), and Cav1 knockout mice exhibited PH with pulmonary vascular remodeling and right ventricular hypertrophy. We then examined roles of Cav1 in human PAECs (hPAECs). Cav1 knockdown in hPAECs reduced phosphorylation of Smad 1/5/9. In addition, Cav1 knockdown significantly increased hypoxia-induced apoptosis in hPAEC. Knockdown of Cavin-1 reversed phosphorylation of Smad 1/5/9 decreased by Cav1 knockdown in BMP9 stimulation. Cavin-1 reversed the expression of BMPRII decreased by overexpression of Cav1. Cav1 was associated with Cavin-1 at the plasma membrane in PAECs. Cav1 also associated with BMPRII at the membrane of hPAECs that was inhibited by Cavin-1, and Cavin-1 reduced the localization of BMPRII to the membrane of hPAECs. These results suggest that BMPRII interacts with Cav1 via Cavin-1-associated localization at the plasma membrane in hPAECs, resulting in regulating BMP/Smad signaling pathway and involving in the development of PAH.

Conclusions

Cavin-1 affects the interaction of Cav1 with BMPRII at the membrane of PAECs, and regulates BMP/Smad signaling. These results reveal a previously undescribed function of Cavin-Caveolin system in the development of PAH through regulation of BMP/Smad signaling.

Funding Acknowledgement

Type of funding source: None
Adipose extracellular signal-regulated kinase 2 protected from endothelial dysfunction and the oxidative stress of perivascular adipose tissue in obese mice
Authors: Osaki A , Sato Y , Endo S , Ito K , Kagami K , Yumita Y , Ishinoda Y , Toya T , Ido Y , Namba T , Adachi T
Published: November 2020

Introduction

Extracellular signal-regulated kinase (ERK) modulates differentiation and maturation of adipocyte and the hypertrophy and differentiation of adipocytes affected the vascular diseases in obese. Changes in characters of adipocytes could develope the oxidative stress and inflammations. Moreover, changes in perivascular adipose tissue (PVAT) could modulate vascular tonus in obesity. However, the role of adipose ERK2 in endothelial function and characters of PVAT in obese in vivo had not been clarified, yet.

Purpose

This study aims to elucidate the role of the adipose ERK2 in endothelial-dependent relaxation (EDR) in mice model of obesity. The role of PVAT in EDR was also assessed.

Methods and results

We created adipose-specific ERK2 knock out mice (AE2KO) by crossing fatty acid binding protein 4 Cre and ERK2 flox mice and fed them with normal diet (ND) or high fat/ high sucrose diet (HFHSD) for 24 weeks. AE2KO fed with HFHSD gained more weight and revealed the heterogeneity in sizes of adipocyte in subcutaneous fat (SF). Furthermore, the mRNA levels of lipoprotein lipase, hormone-sensitive lipase, and peroxisome proliferator-activated receptor γ, which was the master genes of adipocyte differentiation, were markedly down-regulated in SF. PVAT in AE2KO with HFHSD was markedly enlarged and the mRNA expression of inflammatory adipocytokines, such as IL-1β and leptin were up-regulated. Next, we assessed EDR by acetylcholine (ACh) -induced relaxation in aortic rings with or without PVAT. EDR without PVAT was modestly decreased in AE2KO with HFHSD compared with wild type mice (WT) with HFHSD. Aortic rings with PVAT increased EDR in WT with ND. PVAT modestly decreased EDR in WT with HFHSD and mostly eliminated EDR in AE2KO with HFHSD. To assess the contraction factors released from PVAT, the solutions incubated with PVAT (SIP) were transferred to the normal aortic rings. SIP from WT with HFHSD mildly increased vascular tone and SIP from AE2KO with HFHSD further increased it. Tempol, which was superoxide scavenger, restored endothelial dysfunction with PVAT and suppressed the contraction with SIP from AE2KO with HFHSD. Fluorescence intensity of dihydroethidium stain of aorta and PVAT, which indicated that aortic and adipose superoxide production were elevated in AE2KO with HFHSD, which were mostly eliminated with tempol.

Conclusions

Adipose ERK2 selectively modulated differentiation in SF, suppressed the aortic oxidative stress and protected from endothelial dysfunction in obese. Moreover, adipose ERK2 suppressed the hypertrophy, inflammation, and oxidative stress of PVAT in obese. The oxidative stress with the inflammation in PVAT released vasoconstriction factors, which contributed to endothelial dysfunction in obese mice.

Figure 1

Funding Acknowledgement

Type of funding source: None
Myocardial oxygen consumption during histidine-tryptophan-ketoglutarate cardioplegia in young human hearts
Authors: Angeli Emanuela , Martens Sabrina , Careddu Lucio , Petridis Francesco D , Quarti Andrea G , Ciuca Cristina , Balducci Anna , Fabozzo Assunta , Ragni Luca , Donti Andrea , Gargiulo Gaetano D
Published: November 2020

OBJECTIVES

Energy demand and supply need to be balanced to preserve myocardial function during paediatric cardiac surgery. After a latent aerobic period, cardiac cells try to maintain energy production by anaerobic metabolism and by extracting oxygen from the given cardioplegic solution. Myocardial oxygen consumption (MVO2) changes gradually during the administration of cardioplegia.

METHODS

MVO2 was measured during cardioplegic perfusion in patients younger than 6 months of age (group N: neonates; group I: infants), with a body weight less than 10 kg. Histidine-tryptophan-ketoglutarate crystalloid solution was used for myocardial protection and was administered during a 5-min interval. To measure pO2 values during cardioplegic arrest, a sample of the cardioplegic fluid was taken from the inflow line before infusion. Three fluid samples were taken from the coronary venous effluent 1, 3 and 5 min after the onset of cardioplegia administration. MVO2 was calculated using the Fick principle.

RESULTS

The mean age of group N was 0.2 ± 0.09 versus 4.5 ± 1.1 months in group I. The mean weight was 3.1 ± 0.2 versus 5.7 ± 1.6 kg, respectively. MVO2 decreased similarly in both groups (min 1: 0.16 ± 0.07 vs 0.36 ± 0.1 ml/min; min 3: 0.08 ± 0.04 vs 0.17 ± 0.09 ml/min; min 5: 0.05 ± 0.04 vs 0.07 ± 0.05 ml/min).

CONCLUSIONS

We studied MVO2 alterations after aortic cross-clamping and during delivery of cardioplegia in neonates and infants undergoing cardiac surgery. Extended cardioplegic perfusion significantly reduces energy turnover in hearts because the balance procedures are both volume- and above all time-dependent. A reduction in MVO2 indicates the necessity of a prolonged cardioplegic perfusion time to achieve optimized myocardial protection.
The Classification of Extremely Primitive Groups
Authors: Burness Timothy C , Thomas Adam R
Published: November 2020

Dedicated to the memory of Jan Saxl 

Let $G$ be a finite primitive permutation group on a set $\Omega $ with nontrivial point stabilizer $G_{\alpha }$. We say that $G$ is extremely primitive if $G_{\alpha }$ acts primitively on each of its orbits in $\Omega \setminus \{\alpha \}$. These groups arise naturally in several different contexts, and their study can be traced back to work of Manning in the 1920s. In this paper, we determine the almost simple extremely primitive groups with socle an exceptional group of Lie type. By combining this result with earlier work of Burness, Praeger, and Seress, this completes the classification of the almost simple extremely primitive groups. Moreover, in view of results by Mann, Praeger, and Seress, our main theorem gives a complete classification of all finite extremely primitive groups, up to finitely many affine exceptions (and it is conjectured that there are no exceptions). Along the way, we also establish several new results on base sizes for primitive actions of exceptional groups, which may be of independent interest.