bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of albumin on the incidence of adverse events following percutaneous coronary interventions in older adults
Authors: Marti Sanchez D , Duarte Torres J , Marschall A , Biscotti Rodil B , Delgado Calva F , Dejuan Bitria C , Del Castillo Carnevali H , Morales Gallardo M J , Lopez Soberon E , Alvarez Anton S
Published: October 2021

Background

Recent studies have focused on comorbidities or frailty as predictors of adverse events in older adults undergoing percutaneous coronary interventions (PCI), however the prognostic impact of nutritional parameters has been less extensively studied.

Purpose

Our objective was to analyze the impact of serum albumin on the incidence of adverse events after PCI in older patients.

Methods

We conducted a prospective observational study of all-corner consecutive patients aged ≥75 years submitted for PCI at our institution from 2014 to 2017, and with available pre-procedural serum albumin levels. We analyzed the distribution of albumin levels in such population, and the impact of hypoalbuminemia (albumin <3.5 g / dL) on 12-months cumulative incidence of MACE (cardiovascular death, myocardial infarction or stroke), BARC types 3 or 5 major bleeding, and all-cause death.

Results

Study population comprised 169 patients (mean age 82±5 years, 38% women). Mean albumin level was 3.8±0.5 g/dL, and 23% showed levels <3.5 g/dL. Hypoalbuminemia was associated with only mild impairments in baseline cardiac or renal function. During the 12-month follow-up, albumin levels were not associated with a higher incidence of MACE (log-rank p=0.22) but they were associated with a higher incidence of bleeding (22.4% vs. 8.0%, p=0.014), and with all-cause mortality (20.5% vs. 6.2%, p=0.006). Albumin revealed an independent and greater impact than bleeding scores, anemia or discharge anticoagulation on the prediction of bleeding (adjusted HR 2,88, p=0.028).

Conclusions

Albumin levels confer relevant prognostic information that should be assessed in older adults receiving PCI. The effect on bleeding is independent of other well-established variables. Systematic quantification of this widely available parameter may be helpful for clinical decision-making.

Funding Acknowledgement

Type of funding sources: None. Figure 1. Hypoalbuminemia and all-cause death
Less statin associated adverse events after initiation of pitavastatin compared with atorvastatin and rosuvastatin
Authors: Lin J L , Chen P S , Li Y H
Published: October 2021

Background

Statin associated adverse events (SAAEs), including hepatitis, myopathy and new onset diabetes mellitus (NODM), are major reasons that prevent the use of statins. We compared the risk of SAAEs among the commonly used statins to see if SAAEs were similar among the statins.

Methods

We retrieved data from the Taiwan National Health Insurance Research Database. From January 2013 to December 2017, all statin-treated patients without diabetes at baseline were enrolled. We classified eligible patients into pitavastatin (2–4mg/day), moderate-intensity statin (MIS [atorvastatin 10–20 mg/day or rosuvastatin 5–10 mg/day]), and high-intensity statin group (HIS [atorvastatin ≥40 mg/day and rosuvastatin ≥20mg/day]). The study endpoint is a composite of safety events, including hepatitis, myopathy, and NODM. All patients were followed-up for at least one year until December 2018. We used propensity score to balance the baseline differences between the 3 statin groups (N=50935 in each group).

Results

After a mean follow up time of 3.08 years, the safety events occurred in 5014 patients in pitavastatin group (9.84%), 5542 in MIS group (10.88%), and 5343 in HIS group (10.49%). Multivariate Cox proportional hazards model showed that MIS and HIS statins were associated with a higher risk of safety events compared with pitavastatin (adjusted hazard ratio [aHR] 1.122, 95% confidence interval [CI] 1.08–1.17 for MIS and aHR 1.056, 95% CI 1.02–1.10 for HIS). Most events were NODM, with 4818 events in pitavastatin (9.46%), 5284 in MIS (10.37%), and 5113 in HIS group (10.04%). Multivariate Cox proportional hazard analysis showed higher risk of NODM in MIS (adjusted HR 1.111, 95% CI 1.07–1.16) and HIS (aHR 1.050, 95% CI 1.01–1.10) compared with pitavastatin. Time-varying HR analysis showed increased risk of NODM with use of all these statins for more than 1 year compared with non-statin users.

Conclusions

Pitavastatin was associated with a lower risk of SAAEs, especially NODM, compared with atorvastatin and rosuvastatin.

Funding Acknowledgement

Type of funding sources: Private company. Main funding source(s): Orient EuroPharma Co., Ltd.
Comparison of the sensitivity of commercial aPTT tests in measurement of the pharmacodynamic response of milvexian, a novel FXIa inhibitor
Authors: Bonilla F , Bunce M W , Alleman R , Kotha J , Jennings L K , Chintala M
Published: October 2021

Introduction

The activated partial thromboplastin time (aPTT) assay is a test routinely used to evaluate abnormalities or deficiencies in coagulation factors of the intrinsic and common pathways. The composition of the surface activator and phospholipids in the aPTT reagent is known to influence the response of heparin and direct FXa or thrombin inhibitors.

Milvexian (formerly referred to as BMS-986177/JNJ-70033093) is an investigational small-molecule Factor XIa (FXIa) inhibitor being studied for the prevention and treatment of major thrombotic conditions. Clinical pharmacology evaluation of milvexian includes aPTT as the primary pharmacodynamic assay, thus underscoring the need for a sensitive reagent to evaluate its anticoagulant activity.

Purpose

This study evaluated the sensitivity of six commercially available aPTT reagents in plasma from individual donors spiked with milvexian, at concentrations spanning the anticipated clinically relevant exposures.

Methods

Platelet-poor plasma (PPP) prepared from citrated whole blood collected from consenting, healthy adult volunteers (n=12) was spiked with vehicle (DMSO, 0.5% v/v) or 0.1–10 μM milvexian. The aPTT was measured using six commercially available aPTT reagent kits which included silica; kaolin, or ellagic acid as the contact activator, in combination with natural or synthetic phospholipids. The coagulation automated analysers used for testing were matched to the kit's manufacturer recommendations. All reagents and respective normal or abnormal controls were prepared as instructed by the manufacturer.

Results

Milvexian exhibited dose-dependent prolongation of aPTT with all reagents tested. Assays performed with aPTT reagents containing kaolin or ellagic acid demonstrated the highest sensitivity, as measured by the concentration that achieved a 2-fold aPTT prolongation (EC2x), and showed the widest dynamic range of response. Coefficient of variability of aPTT measurements in plasma from 12 individual donors was between 5.6–7.9% for Dade® Actin® FS, and 7.1–8% for STA®-C.K. Prest® 5.

Conclusions

Prolongation of aPTT in PPP spiked with milvexian exhibited a dose-dependent relationship, with statistically significant differences observed among reagents at milvexian concentrations above 0.3 μM. The highest sensitivity measured by changes in the ratio to baseline was obtained with aPTT reagents containing an activator/procoagulant phospholipid combination of ellagic acid + purified soy phosphatides or kaolin + cephalin, which performed similarly. Identification of the reagents with the best combination of sensitivity, precision, and dynamic range may help guide the selection of reagent for assessing milvexian activity.

Funding Acknowledgement

Type of funding sources: Private company. Main funding source(s): Janssen Research & Development, LLC
Efficacy and safety of direct oral anticoagulants with diabetes and nonvalvular atrial fibrillation: a systematic review and meta-analysis
Authors: Costa G , Goncalves L , Teixeira R
Published: October 2021

Background

Diabetes Mellitus (DM) is an independent risk factor for stroke and atrial fibrillation (AF). Therefore, the risk/benefit profile of the direct oral anticoagulants (DOAC) is of clinical interest.

Purpose

To compare efficacy and safety outcomes of DOAC for nonvalvular AF in patients with DM versus without DM.

Methods

We systematically searched PubMed, Embase and Cochrane databases, in January 2020, for interventional studies comparing DOAC efficacy and safety in patients with AF and diabetes versus without diabetes.

Results

Four randomized clinical trials were included, providing a total of 63987 patients, 18860 with DM and 45127 without DM. In terms of efficacy, our meta-analysis revealed a similar rate of stroke/systemic embolism (pooled OR 1.02 [0.79, 1.31], P=0.87, I2=83%), stroke (pooled OR 1.98 [0.68, 1.40], P=0.90, I2=90%) and all-cause mortality (pooled OR 1.18 [0.97, 1.43], P=0.10, I2=87%), albeit with a significant heterogeneity. However, in direct factor Xa inhibitors sub analysis, diabetic patients had a lower trend of systemic embolism/stroke (pooled OR 0.90 [0.79, 1.02], P=0.09, I2=18%), significantly lower stroke rate (pooled OR 0.82 [0.73, 0.93], P<0.01, I2=0%), but a higher all-cause mortality (pooled OR 1.08 [1.00, 1.16], P<0.01, I2=0%). In terms of safety, the diabetic patients receiving DOAC had higher rates of major bleeding events (pooled OR 1.28 [1.14, 1.45], P<0.01, I2=50%), although with significant heterogeneity. Direct factor Xa inhibitors sub analysis also revealed a higher rate of major bleeding events (pooled OR 1.22 [1.08, 1.38], P<0.01, I2=24%), but a similar intracranial bleeding events (pooled OR 1.03 [0.86, 1.24], P=0.72, I2=0%).

Conclusion

Our pooled analysis suggests that diabetic patients on DOAC have an higher bleeding risk on DOAC, although with a superior embolic protection.

Funding Acknowledgement

Type of funding sources: None. Systemic Embolism/Stroke in DM vs. NonDM
Detection of myocardial ischemia by intracoronary ECG using convolutional neural networks
Authors: Bigler M R , Seiler C
Published: October 2021

Introduction

The electrocardiogram (ECG) is a valuable tool for the diagnosis of myocardial ischemia as it presents distinctive ischemic patterns. Deep learning methods such as convolutional neural networks (CNN) are employed to extract data-derived features and to recognize natural patterns. This study aimed to determine the optimal ischemic parameter as obtained from the highly susceptible intracoronary ECG (icECG) using pre-trained CNN.

Method

This was a retrospective observational study in 228 patients with chronic coronary syndrome. Each patient had participated in clinical trials with icECG recording and ST-segment shift measurement at the beginning (i.e.,non-ischemic) and the end (i.e.,ischemic) of a one-minute proximal coronary artery balloon occlusion establishing the reference. Using these data (893icECGs in total), two pre-trained, open-access CNN (GoogLeNet/ResNet101) were trained to recognize ischemia. The best performing CNN during training were compared with the icECG ST-segment shift for diagnostic accuracy in the detection of artificially induced myocardial ischemia.

Results

Using coronary patency or occlusion as reference for absent or present myocardial ischemia, receiver-operating-characteristics (ROC)-analysis of manually obtained icECG ST-segment shift (mV) showed an area under the ROC-curve (AUC) of 0.903±0.043 (p<0.0001, sensitivity 80%, specificity 92% at a cut-off of 0.279mV). The best performing CNN showed an AUC of 0.924 (sensitivity 93%, specificity 92%). DeLong-Test of the ROC-curves showed no significant difference between the AUCs. The underlying morphology responsible for the network prediction differed between the trained networks but was focused on the ST-segment and the T-wave for myocardial ischemia detection.

Conclusions

When tested in an experimental setting with artificially induced coronary artery occlusion, quantitative icECG ST-segment shift and CNN using pathophysiologic prediction criteria detect myocardial ischemia with similarly high accuracy.

Funding Acknowledgement

Type of funding sources: None.
Triage-HF plus: 12-month study of remote monitoring pathway for triage of heart failure risk initiated during the Covid-19 pandemic
Authors: Bachtiger P , Park S , Letchford E , Scott F , Barton C , Ahmed F Z , Cole G , Keene D , Plymen C M , Peters N S
Published: October 2021

Background

The Covid-19 pandemic necessitated rapid adoption of remote monitoring across cardiovascular patient cohorts. Most patients with cardiac implantable electronic devices (CIEDs) are now able to be remotely monitored using either scheduled, patient- or threshold-triggered transmissions. The validated “Triage Heart Failure Risk Score” (Triage-HFRS) is a medical algorithm within company-specific CIEDs that can risk-stratify patients as low-, medium- or high-risk of worsening heart failure (WHF) in the next 30 days based on integrated monitoring of physiological parameters. Building on a previous proof-of-concept of the Triage-HF Plus pathway, we integrated remote data with simple 5-question telephone triage within a clinical pathway to identify WHF during the first year of the Covid-19 pandemic.

Purpose

Prospective evaluation of clinical remote monitoring pathway integrating Triage-HFRS with protocolised telephone triage (Triage-HF Plus pathway).

Methods

Prospective, real-world evaluation of clinical pathway serving a large urban region over a 12-month period, using data from April 2020 to April 2021 (initiated during the first wave of Covid-19 pandemic in the UK). From a population of 435 patients with CIEDs, 87 “high” Triage-HFRS alerts were received and patients contacted for telephone triage assessment. Screening questions were designed to identify episodes of WHF and non-HF events. Intervention was at discretion of the clinical practitioner and in line with guideline-directed practice. A consecutive sample of 115 “medium” risk scores received the same triage.

Results

Successful contact was made with 72 (82.8%) high-risk patients. Classification for high scoring patients confirmed on triage included isolated heart failure (18.3%), heart failure concurrent to medical problem (5.7%), alternative medical problem (10.3%), and recent hospital admission (8.0%); triage reassured absence of acute cause of high score in 40.2%. The sensitivity and specificity for detection of WHF was 87.9% (0.77–0.99) and 59.4% (0.50–0.69) respectively. Positive and negative predictive values were 40.3% and 94.0%, respectively. Overall accuracy was 66.2%.

Conclusions

The Triage-HF Plus pathway served as a useful remote monitoring tool for identifying patients with WHF whose care had been otherwise disrupted by the Covid-19 pandemic, allowing timely intervention and cementing the longer-term role for such models of care delivery. Crucially, in this multimorbid, high-cost population, relevant non-HF issues were also identified. The high negative predictive value further highlights the potential of proactive surveillance over conventional, periodic follow up.

Funding Acknowledgement

Type of funding sources: Foundation. Main funding source(s): Imperial Health Charity
Sex-related differential susceptibility to ponatinib-induced cardiotoxicity and its relationship to modulation of Notch signaling in a murine model
Authors: Madonna R , Pieragostino D , Cufaro M C , Del Boccio P , Pucci A , Mattii L , Doria V , Cadeddu Dessalvi C , Zucchi R , Mercuro G , De Caterina R
Published: October 2021

Background

Ponatinib (PON), a tyrosine kinase inhibitor approved in chronic myeloid leukemia, has proven cardiovascular toxicity. Although sex is a risk factor for PON-induced cardiotoxicity in humans, little is known about its mechanisms in general, and sex-related mechanisms in particular.

Objectives

To determine the mechanisms of sex-related PON-induced cardiotoxicity and identify potential rescue strategies in a murine model.

Methods

24-months-old male and female C57B5 mice were treated with 3 mg/kg/day of PON or vehicle via oral gavage for 28 days, with/without siRNA-Notch1 or siRNA-scrambled via tail vein every 3 days.

Results

PON + scrambled siRNA-treated male mice had a higher number of TUNEL-positive cells, a higher percentage of senescence-associated β-galactosidase positive senescent cardiac areas, as well as a lower reactivity degree for the survival marker Bmi1 than female counterparts. Proteomics analysis of cardiac tissue showed upstream activation of nitric oxide synthase (NOS) type 2, downstream activation of cell death and production of reactive oxygen species in PON + scrambled siRNA- compared to vehicle or PON + Notch1 siRNA-treated male mice. Upstream analysis showed beta-estradiol activation, while downstream analysis showed activation of cell survival and inhibition of cell death in PON + scrambled siRNA compared to vehicle-treated female mice. PON + scrambled siRNA-treated mice also showed a downregulation of cardiac actin, which was more marked in male; as well as vessel density, which was more marked in female mice. Female hearts showed a greater extent of cardiac fibrosis than male counterparts at baseline, with no significant changes after PON treatment. In contrast, PON + scrambled siRNA-treated mice had less fibrosis than vehicle or PON + Notch1-siRNA-treated mice. Left ventricular systolic dysfunction shown in PON + scrambled siRNA-treated male mice and - to a lesser extent - by female mice was similarly reversed in both PON + Notch1-siRNA-treated male and female mice (Table 1).

Conclusions

We found a sex-related differential susceptibility and Notch1 modulation in PON-induced cardiotoxicity. This can improve our understanding of sex-related differences and help identify biomarkers in PON cardiotoxicity.

Funding Acknowledgement

Type of funding sources: None.
Genome-wide association study of atrial fibrillation in 114,539 Finnish individuals reveals novel locus associated with cardiac remodelling
Authors: Paludan-Muller C , Vad O B , Svendsen J H , Olesen M S
Published: October 2021

Background/Introduction

Atrial fibrillation (AF) is the most common cardiac arrhythmia and it is associated with serious complications, such as stroke, heart failure, and premature death. Previous genome-wide association studies (GWAS) have associated more than 140 genomic loci with AF; however, these studies predominantly include subjects of European ancestry. Although, the Finnish population is European, it is genetically considered different from other European populations as it has been isolated and developed through multiple bottlenecks followed by population growth. Therefore, pathogenic variants are more easily discovered and heritably diseases are more prevalent.

Methods

We accessed summary statistics on atrial fibrillation and flutter (I48) from the Finngen project. Loci were defined as 1 megabase regions around lead SNPs, and loci were considered novel when the SNPs had P-values <5x10–8 after conditional analysis, and no previously reported SNPs were within the loci. FINEMAP was done with a Finnish LD reference panel, and colocalization of GWAS and eQTL signals were analysed with MetaXcan.

Results

A GWAS on 17,325 Finnish AF cases and 97,214 controls confirms 16 previous identified loci and reveals one novel locus on chromosome 19. The novel lead SNP, rs190065070 (odds ratio [OR] = 1.44, 95% confidence interval [CI] = 1.29–1.61, P-value = 5.96x10–11), is close to the gene EMC10, which encodes the endoplasmic reticulum membrane protein complex subunit 10. While the locus harbours other genes, our MetaXcan analysis could not provide conclusive evidence for other plausible genes. The EMC complex consists of 10 subunits and is a chaperone in endoplasmic reticulum-resident membrane proteins. Previous mouse studies have shown EMC10 to be important in angiogenesis after myocardial infarction, and it has recently been associated with a novel neurodevelopment syndrome. The EMC1 subunit has been associated with congenital heart disease.

Conclusion

We present a novel susceptibility locus associated with AF in the Finnish population. The locus is in proximity to the gene EMC10, which is involved in structural remodelling of the heart after myocardial infarction. These results propose a potentially novel pathophysiological pathway in AF.

Funding Acknowledgement

Type of funding sources: Public hospital(s). Main funding source(s): The Research Foundation RigshospitaletThe John and Birthe Meyer Foundation
Morphological features of mitral annular calcification leading to systolic anterior motion of the mitral valve
Authors: Friend E J , Wiener P C , Murthy K , Pressman G S
Published: October 2021

Background

Systolic anterior motion (SAM) of the mitral valve (MV) can develop after mitral valve repair with placement of an annuloplasty ring. It is occasionally seen in patients with mitral annular calcification (MAC) but mechanisms have not been carefully delineated. Using 2-dimensional echocardiography we explored morphologic parameters which may contribute to SAM in patients with MAC.

Hypothesis

We hypothesized that in cases of MAC where SAM is present there would be anterior displacement of the valve by the posterior annular calcification.

Methods

From our echocardiographic database we identified 20 patients with severe MAC who also had SAM with definite septal contact. Each subject was paired with 2 controls free of MAC and 1 control with severe MAC but no SAM. All controls were matched for age, sex, BSA, and septal wall thickness (±1.5 mm). 2-D echocardiographic measurements were taken from the parasternal long-axis (PLAX), apical 3-chamber and apical 4-chamber views.

Results

MAC+SAM vs MAC no-SAM. Three notable differences were observed: MAC+SAM patients, as compared with MAC no-SAM, had a smaller left ventricular outflow tract (LVOT), longer anterior mitral leaflet, and greater displacement of the MV coaptation point towards the interventricular septum (Figure 1). Median values for these 3 factors were determined using the no-MAC controls; each MAC subject was then scored for number of factors exceeding those values. MAC+SAM patients had a mean score of 2.7 vs 1.1 for MAC no-SAM patients. By combining anterior mitral leaflet length and coaptation point-septal distance as a ratio we could effectively separate MAC+SAM vs MAC no-SAM when >0.9 with one exception (Figure 2). We also observed a smaller anteroposterior annular dimension in the MAC+SAM group.

MAC no-SAM vs no-MAC. Comparing these groups there were no differences in LVOT diameter or coaptation-septal distance; effective anterior mitral leaflet length was smaller in MAC no-SAM subjects vs no-MAC controls while anteroposterior dimension of the annulus was larger.

Conclusions

SAM develops in a subset of patients with severe calcification of the mitral annulus. These patients have a smaller anteroposterior annular dimension, possibly due to severe MAC. Other notable differences characterize MAC patients with SAM from those without. The LVOT is smaller, the effective anterior mitral leaflet length is longer, and the point of leaflet coaptation is displaced towards the septum. Using the ratio of anterior mitral leaflet length/coaptation point-septal distance in this study sample effectively separated those MAC patients with SAM from those without.

Funding Acknowledgement

Type of funding sources: None. Figure 1Figure 2
Epicardial adipose tissie-derived leptin induce MMPs/TIMPs imbalance and promotes myocardial remodeling in high fat diet-induced obese rats
Authors: Tian G , Liu L , Luo C D , Li M , Cao L F , Zhu D J
Published: October 2021

Background and aims

Epicardial adipose tissue (EAT)-derived leptin contributes to myocardial remodeling in metabolic syndrome. However, the precise mechanisms remain to be determined. The present study was designed to elucidate the adverse effects of EAT-derived leptin on obesity-related myocardial remodeling.

Methods and results

Eight-week-old male Wistar rats were divided into two groups that received either a normal diet (control, n=10) or a high-fat diet (obese, n=10) for 12 weeks. Obese rats exhibited abnormal myocardial structure, diastolic dysfunction and abundant collagen deposition. Local leptin expression in obese rats EAT upregulated along with adipocyte hypertrophy, accompanied by renin-angiotensin-aldosterone system (RAAS) activation and increased oxidative stress level. Leptin receptor (ObR) and angiotensin II type 1 receptor (AT1R) expression in obese EAT were significantly higher than that in control. In vitro, mature adipocytes treated with angiotensin II (Ang II) exhibited pronounced leptin synthesis and secretion by promoting AP-1 nuclear translocation via the AT1R-ROS-ERK1/2 pathway. Moreover, cardiac fibroblasts were incubated with obese rat EAT-conditioned medium (EAT-CM), plus various inhibitors. EAT-derived leptin promoted proliferation of cardiac fibroblasts associated with increased ERK1/2 phosphorylation and induced MMPs/TIMPs imbalance, stimulating upregulation of type I collagen via the JAK2/STAT3-TGF-β1/Smad3 pathway in cardiac fibroblasts of obese rats.

Conclusions

The paracrine effect of EAT-derived leptin on myocardial remodeling, inducing MMPs/TIMPs imbalance and promoting the proliferation of cardiac fibroblasts via activating ERK1/2 and JAK2/STAT3-TGF-β1/Smad3 pathway in obesity.

Funding Acknowledgement

Type of funding sources: Foundation. Main funding source(s): the Nature Science Foundation of China (grant no. 81873513, 81600574, and 30871042)
Identification of miRNA-497 and miRNA-27b-5p as potential diagnostic markers of cardiac fibrosis
Authors: Tikhomirov R , Reilly-O'donnell B , Lucarelli C , Greco S , Zacagnini G , Maryam A , Menicanti L , Leszek P , Faggian G , Srivastava P , Emanueli C , Martelli F , Gorelik J
Published: October 2021

Background

Cardiac fibrosis is associated with inflammation and extracellular matrix (ECM) accumulation. A pro-fibrotic cytokine, IL11 induces cardiac fibroblasts conversion to myofibroblasts expressing α-smooth muscle actin (α-SMA) and ECM. MicroRNAs (miRNAs) are a class of small non-coding RNAs which participate in regulation of gene expression; Although mainly intracellular, miRNAs can be released into the blood stream where they can be readily detected.

Purpose

To screen miRNAs upregulated following IL11 triggered conversion of rat cardiac fibroblasts into myofibroblasts. To validate these miRNAs as potential diagnostic biomarkers of cardiac fibrosis by testing their level in blood plasma and septum of aortic valve stenosis (AVS) patients.

Methods and results

With a bioinformatical approach (Figure 1), we predicted miRNAs which can target proteins involved in TGFβ and IL-11 pathways of fibrosis progression. Of a vast number of miRNAs, we identified 7 strong candidates. After qPCR validation, we found miRNA-27b-5p and miRNA-497 to be significantly upregulated in rat cardiac fibroblasts treated by IL11 (5 ng/ul) but not TGFβ1 (100 ng/ul), values are 2–ΔΔCt: (3±1.5) and (5.2±2.2) (p-value <0.05) for miRNA-27b-5p and miRNA-497 respectively. Next, we overexpressed these two miRNAs separately in rat cardiac fibroblasts. With immunostaining we observed a (18.3±6.8)% increase in the percentage of α-SMA positive cells for miR-27b-5p and a (38.0±8.3)% increase for miR-497. Moreover, we detected with qPCR a significant up-regulation of α-SMA expression (−ΔΔCt = 3.4±0.9 for miR-27b-5p; −ΔΔCt = 8.2±0.7 for miR-497) in cells overexpressing miRNA27b-5p and miRNA497. Furthermore, we found that levels of both miRNA-27b-5p and miRNA-497 were significantly higher in blood plasma (p=0.0002, p=0.04) of AVS patients compared to age and sex matched control group of healthy donors (Figure 2) and heart septum (p=0.0004, p=0.04) of AVS patients compared to septum of healthy donors that could not be used for transplantation. In addition, quantification of Sirius red staining and immunohistochemistry for Col1a1 displayed significant ECM accumulation in AVS patients (p=0.04).

Conclusions

We found miRNA-497 and miRNA-27b-5p to be pro-fibrotic in rat fibroblasts. Importantly, we found both miRNAs to be up-regulated in the peripheral blood of AVS patients.

Funding Acknowledgement

Type of funding sources: Other. Main funding source(s): Roman Tikhomirov PhD studentship is supported by a fellowship from the University of Verona, ItalyEEU-Cardiac RNA cost action CA17129
Epigenetic regulation of oxidative and inflammatory phenotypes in women with gestational diabetes and offspring
Authors: Di Pietrantonio N , Shumliakivska M , Suades R , Di Tomo P , Bonfini T , Pandolfi A , Cosentino F
Published: October 2021

Background

Hyperglycemia-induced oxidative stress and inflammation are potent drivers of atherosclerotic cardiovascular disease (ASCVD). Gestational diabetes (GDM) is characterized by chronic hyperglycemia during pregnancy and may represent a clinical model to study the mechanisms of oxidative stress and inflammation induced by hyperglycemia. GDM is associated with a range of adverse perinatal and long-term outcomes for both mother and offspring. In this perspective, it is emerging a putative association between maternal GDM and offspring's epigenetic trait.

Purpose

To investigate the link between histone modifications, oxidative stress and inflammatory phenotype as well as the transmission of epigenetic signatures to the offspring.

Methods

We analyzed peripheral blood mononuclear cells (PBMC) from GDM and control mothers as well as human umbilical vein endothelial cells (HUVEC) and cord blood mononuclear cells (CBMC) isolated from newborn umbilical cords obtained at delivery from both groups. Histone methyltransferase MLL1-dependent trimethylation of histone 3 at lysine 4 amino residue (H3K4me3) on NF-kB p65 subunit promoter region was assessed by chromatin immunoprecipitation (ChIP) and real-time qPCR in HUVEC, PBMC and CBMC, respectively. MLL1 and downstream inflammatory and redox genes were determined by real-time qPCR and immunocytochemistry in the presence and in the absence of MLL1 inhibitor MM-102. Measurement of reactive oxygen species (ROS) was performed by electron spin resonance spectroscopy.

Results

For the first time, we demonstrated a significant increase of MLL1 expression with subsequent MLL1-induced upregulation of NF-kB p65 gene via H3K4me3 in GDM as compared to control cells. MLL1-driven epigenetic remodeling of NF-kB p65 promoter is upstream to the activation of inflammatory pathway. Indeed, treatment with MM-102 decreased H3K4me3 and blunted expression of NF-kB p65 as well as VCAM-1, MCP-1 and IL-6 genes. We also found that expression of ROS scavenger aldehyde dehydrogenase 2 is reduced, whereas pro-oxidant nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunit NOX4 is upregulated. Interestingly, the increased ROS generation observed in GDM is involved in the upregulation of MLL1 as shown by the restoring effect of antioxidant vitamin C on MLL1 expression levels.

Conclusions

Our results suggest that a complex interplay between oxidative stress and histone modifications are responsible for the GDM maternal inflammatory and oxidative phenotypes and its transmission to the offspring. The deciphering of epigenetic-induced chromatin remodelling opens the perspective for pharmacological reprogramming of adverse chromatin changes to reduce the burden of early development of metabolic phenotypes and ASCVD.

Funding Acknowledgement

Type of funding sources: Public Institution(s). Main funding source(s): University G. d'Annunzio MIUR fundings Schematic figure
Proatherogenic gut-microbiome metabolite trimethylamine elicits a gut-liver circuit to regulate TMAO metabolism and atherosclerosis via mediation of CREBH
Authors: Su Q , Kaluzny S
Published: October 2021

Introduction

In humans, circulating metabolite Trimethylamine N-oxide (TMAO) is closely associated with higher risk of cardiovascular disease. Trimethylamine (TMA), a precursor of TMAO, is produced by gut microbiome using dietary components, i.e., choline and carnitine, as substrates. The gut-derived TMA is then transferred to the liver where it is further oxidized to TMAO by the flavin-containing monooxygenases (FMOs). The ER-resident transcription factor c-AMP responsive element binding protein H (CREBH/CREB3L3) is exclusively expressed in the liver and intestine. Perturbation of CREBH activity contributes to the development of hyperlipidemia and cardiovascular disease. Therefore, the Purpose of this study is to investigate the regulatory effect of a gut bacterium, Akkermansia muciniphila (A. muciniphila), on TMA and TMAO metabolism and the role of CREBH in this process.

Methods

Two groups of wild type (WT) and CREBH knockout (CREBH-KO) mice were inoculated with 200 μL of A. muciniphila (2×108 cfu/0.2 mL) in PBS or the vehicle (PBS) alone as control every other day through oral gavage for 2 weeks. Plasmas, liver and intestinal tissues were collected for metabolomics analysis, immunoblotting analysis and q-RT-PCR.

Results

Metabolomics analysis of the plasmas from the experimental mice revealed that increased colonization of A. muciniphila in the gut significantly reduced circulating TMA in the WT mice but not in CREBH-KO mice (P<0.05), suggesting that depletion of CREBH altered the microenvironment of gut microbiome which affected the metabolism of TMA by gut bacteria. In the livers, A. muciniphila treatment markedly reduced mRNA expression of FMO1 and FMO3 (P<0.05), which subsequently inhibited the enzymatic conversion of TMA to TMAO in hepatocytes. Immunoblotting analysis further revealed that LDL receptor was upregulated whereas ER stress markers, GRP94 and JNK1/2, were downregulated in the A. muciniphila treated KO mice, indicating an acceleration in lipoprotein (VLDL remnant) clearance from the circulation and the improvement of metabolic inflammation. In vitro, incubation of mouse hepatocytes AML12 with TMA (600 mM) for 12 hours stimulated expression of FMOs to facilitate the conversion of TMA to TMAO and induced lipotoxicity.

Conclusion

CREBH mediates the crosstalk between gut microbiome and liver metabolic system that regulates TMA and TMAO metabolism, which contributes to the induction of metabolic inflammation and atherogenesis. This novel finding may lend support to the therapeutic strategy of atherosclerosis.

Funding Acknowledgement

Type of funding sources: Foundation. Main funding source(s): British Heart Foundation
Eicosapentaenoic acid inhibits lipopolysaccharide (LPS)-induced nitrite production and cytokine release from J774 macrophages
Authors: Sherratt S C R , Libby P , Dawoud H , Bhatt D L , Malinski T , Mason R P
Published: October 2021

Background

Eicosapentaenoic acid (EPA), an omega-3 (ω-3) fatty acid, reduced cardiovascular (CV) events in high-risk patients (REDUCE-IT) but the mechanism is not fully understood. Activated macrophages, characterized by cytokine release and increased inducible nitric oxide synthase (iNOS) activity, contribute to atherosclerosis. As both a substrate for and potential inhibitor of cyclooxygenase (COX), EPA may reduce iNOS activity.

Purpose

The purpose of this study was to evaluate the dose-dependent effects of EPA on nitrite and cytokine release from lipopolysaccharide (LPS)-activated macrophages.

Methods

Murine J774 macrophages were pretreated with vehicle or EPA at 10, 20 and 40 μM for 2 h, then challenged with LPS at 1.0 μg/ml. After 24 hr, iNOS activity was measured by nitrite production using the Griess assay. EPA was compared to the COX inhibitor diclofenac at 1.0 μg/ml. Levels of interleukin-1β (IL-1β) and tumor necrosis factor alpha (TNF-α) in cell supernatant were measured by immunochemistry using colchicine as a positive control.

Results

Activated macrophages caused a >4-fold increase in nitrite production (p<0.001) that was reduced by EPA in a dose-dependent manner. EPA decreased nitrite levels by 40, 62 and 77% at 10, 20 and 40 μM, respectively (p<0.01). Diclofenac separately reduced nitrite levels by 40% (p<0.01). EPA also reduced expression of IL-1β and TNF-α by 40% and 31%, respectively (p<0.01), in a manner similar to equimolar colchicine (10 μM). The reductions in IL-1β and TNF-α with EPA were dose-dependent.

Conclusions

EPA reduced macrophage activation as evidenced by decreased nitrite production and cytokine release similar to other anti-inflammatory agents. These findings indicate a novel effect of EPA on mechanisms of inflammation associated with vascular disease.

Funding Acknowledgement

Type of funding sources: Private company. Main funding source(s): Amarin Pharma Inc., Elucida Research
Tubulin-folding cofactor E deficiency is associated with vascular dysfunction and endoplasmatic reticulum stress of vascular smooth muscle cells
Authors: Efentakis P , Molitor M , Kossmann S , Bochenek M , Wild J , Lagrange J , Finger S , Jung R , Karbach S , Schaefer K , Schulz A , Wild P , Muenzel T , Wenzel P
Published: October 2021

Introduction

Endothelial function assessed via flow mediated dilatation (FMD) has shown to predict risk in individuals with established cardiovascular diseases, whereas its predictive value is uncertain in the setting primary prevention.

Purpose

The aim of the current work was to discover and evaluate novel mediators of vascular dysfunction in the general population and in conditional knock-out transgenic animal models.

Methods

In order to identify novel targets that were negatively correlated with FMD and investigate their contribution in vascular function, a Genome Wide Association Study (GWAS) of 5,000 participants was performed and subsequently immune cell-, endothelial- and vascular smooth muscle cell (VSMCs)-targeted conditional knockout mouse models were generated and characterized.

Results

GWAS analysis revealed that single nucleotide polymorphisms (SNPs) in the tubulin folding cofactor E (TBCE) gene were negatively correlated with FMD and TBCE expression in the peripheral blood mononuclear cells (PBMCs). Myelomonocytic cell-targeted TBCE deficiency did not lead to any vascular dysfunction in vivo in the LysM+Cre+/−TBCEfl/fl mice. Endothelial-targeted TBCE deficiency led to an NLR family pyrin domain containing 3 (NLRP3)-dependent activation of the inflammasome in the endothelial cells of Tie2-ERT2Cre+/−TBCEfl/fl mice. Importantly, VSMC-targeted TBCE deficiency was associated with endothelial dysfunction, increased aortic wall thickness and endoplasmatic reticulum (ER) stress-mediated VSMC hyperproliferation in vivo (SMMHC-ERT2Cre+/−TBCEfl/fl), paralleled by calnexin upregulation. Administration of the blood pressure hormone angiotensin II exacerbated the vascular dysfunction and phenotype. Administration of the ER stress modulator tauroursodeoxycholic acid to the SMMHC-ERT2Cre+/−TBCEfl/fl mice reversed vascular dysfunction, paralleled by induction of Raptor/Beclin-1-dependent autophagy both in vitro and in vivo.

Conclusion

TBCE and tubulin homeostasis in the vascular musculature seem to be novel markers of vascular function and represent a new druggable target for the treatment of ER-stress-mediated vascular dysfunction.

Funding Acknowledgement

Type of funding sources: Foundation. Main funding source(s): This work was supported by grants of the German Federal Ministry for Education and Research (BMBF01EO1003 and BMBF01EO1503), the DFG Major Research Instrumentation Programme (DFG INST 371/47-1 FUGG) as well as the Boehringer Ingelheim Foundation. PW received funds from the German Research Foundation in support of his work (DFG WE4361-4-1 and WE 4361/7-1). KS, TM and PW are PIs of the DZHK.
Assessment of heart rate variability and tone of the autonomic nervous system in puberty-aged children with undifferentiated connective tissue dysplasia
Authors: Sidorovich O V , Elizarova S Y U , Khizhnyak A V , Kuznetsova N A , Prosova E E
Published: October 2021

Within the framework of the research into pathological conditions and syndromes associated with undifferentiated connective tissue dysplasia (UCTD), an emphasis needs to be placed on the changes in heart rhythm. In puberty, the severity of neurovegetative disorders caused by the presence of UCTD might increase manifold. The features of heart rate variability (HRV) in pubertal children against the background of UCTD have been studied insufficiently and require more detailed analysis.

This study aims to assess heart rate variability and tone of the autonomic nervous system (ANS) in puberty-aged children with UCTD.

Two groups of children were examined, with methods including electrocardiography and cardiointervalography. The main group consisted of 36 puberty-aged children with UCTD, the control group consisted of 30 healthy children.

ANS tone was assessed based on the HRV. In the control group of children aged 11, the value of variation range (VR) was 0.18±0.4, mode amplitude (MA) 35.2±2.4, stress index (IN) 72.8±4.3, which corresponds to the state of vegetative balance. As the puberty progresses, the state of eutonia is replaced by the activation of the sympathetic division of the ANS, which becomes most pronounced at the age of 13 years. In the control group of children aged 13, the value of VR was 0.08±0.02, MA 61.7±2.1, SI 210.6±8.6, which corresponds to moderate sympathicotonia. By the end of puberty, in healthy children, the severity of sympathetic activation begins to decrease, and at the age of 15 years, vegetative balance is again observed. However, this pattern is violated in children with UCTD. Upon entering puberty, moderate activation of the sympathetic division of the ANS is already observed in children with UCTD. At the age of 11 years in this group, the value of VR is 0.06±0.02, MA 72.4±3.1, SI 211.2±8.6. At the age of 13, pronounced sympathicotonia is observed, which is confirmed by the values of VR - 0.04±0.01, MA 80.2±4.3 and SI 34.3±14.7. By the end of puberty in children with UCTD, in contrast to the control group, the analyzed parameters indicate not an equilibrium of the ANS, but the predominance of parasympathetic activation.

The research provides an insight into heart rate variability in puberty-aged children with UCTD. It was established that healthy children enter puberty in a state of equilibrium of the autonomic nervous system, which in the middle of puberty is replaced by moderate sympathicotonia, and then returns to its original values. Children with the presence of UCTD enter puberty with moderately pronounced sympathicotonia, which gradually increases to pronounced values and is replaced by moderate vagotonia.

Funding Acknowledgement

Type of funding sources: None.
A resilient type of familial hypercholesterolaemia: case–control follow-up of genetically characterized older patients in the SAFEHEART cohort
Authors: Pérez de Isla Leopoldo , Watts Gerald F , Muñiz-Grijalvo Ovidio , Díaz-Díaz Jose Luis , Alonso Rodrigo , Zambón Daniel , Fuentes-Jimenez Francisco , Mauri Marta , Padró Teresa , Vidal-Pardo José I , Barba Miguel A , Ruiz-Pérez Enrique , Michán Alfredo , Mediavilla Juan D , Hernandez Antonio M , Romero-Jimenez Manuel J , Badimon Lina , Mata Pedro ,   SAFEHEART Investigators
Published: October 2021

Aims 

Knowledge of the features of patients with familial hypercholesterolaemia (FH) who are protected from atherosclerotic cardiovascular disease (ASCVD) is important for the clinical and prognostic care of this apparently high-risk condition. Our aim was to investigate the determinant and characteristics of patients with FH who are protected from ASCVD and have normal life expectancy, so-called ‘resilient’ FH (R-FH).

Methods and results 

Spanish Familial Hypercholesterolaemia cohort study (SAFEHEART) is an open, multicentre, nation-wide, long-term prospective cohort study in genetically defined patients with heterozygous FH in Spain. Patients in the registry who at the time of analysis were at least 65 years or those who would have reached that age had they not died from an ASCVD event were analysed as a case–control study. Resilient FH was defined as the presence of a pathogenic mutation causative of FH in a patient aged ≥65 years without clinical ASCVD. Nine hundred and thirty registrants with FH met the study criteria. A defective low-density lipoprotein (LDL)-receptor mutation, higher plasma level of high-density lipoprotein cholesterol (HDL-C), younger age, female gender, absence of hypertension, and lower plasma lipoprotein (a) [Lp(a)] concentration were independently predictive of R-FH. In a second model, higher levels of HDL-C and lower 10-year score in SAFEHEART-RE were also independently predictive of R-FH.

Conclusion 

Resilient FH may be typified as being female and having a defective LDL-receptor mutation, higher levels of plasma HDL-C, lower levels of Lp(a), and an absence of hypertension. The implications of this type of FH for clinical practice guidelines and the value for service design and optional care of FH remains to be established.

Trial registration

ClinicalTrials.gov number NCT02693548.
COVID vaccination efficiency in nursing homes: from nursing home-centred to resident-centred
Authors: Delespierre T , Pierre A
Published: October 2021

Background

Korian® group is specialized in care and support for fragile people. Its data warehouse manages 304 French nursing homes (NH). The transmissions' table (TT), daily-fed by caregivers, contains key data about residents' care and health. Following GDPR rules, we could still build residents' life trajectories and detail all the residents' health events using indexes. In the past, we designed syndromes such as falls, flu, and last year, COVID with the standard query language. Now we want to combine ‘hard' data such as sociodemographic profiles and COVID vaccines with ‘soft' textual data such as COVID and emergency calls syndromes to get a better residents vaccine monitoring.

Methods

First, we selected all residents with at least one transmission, from December 21, to January 11, just before the COVID vaccination start, from 34 NH chosen for their variety of size and geographic location. We followed them for eleven weeks starting January 4, 2021. We selected the residents' age, sex, autonomy level, comorbidities and risks and then, vaccinations (0, 1 or 2), syndromes, hospitalizations and deaths date events. We searched for sample data inconsistencies by filtering COVID textual information in the TT and comparing it with the vaccination table. We described the sample according to the NH size, and then according to the number of vaccinations. Finally, we assessed COVID syndromes, falls, hospitalizations, emergency calls and deaths according to vaccinations followed by Poisson and logistic regressions.

Results

We built a 2,051 residents cohort with 1,401 vaccinated at least once and 650 not yet vaccinated. Data corrections then added 60 vaccinations. Residents in big structures were more masculine (72,9% vs 75,1%), more vaccinated (77,2% vs 67,2%), younger (86,7 vs 87,5), more autonomous (2,6, vs 2,4). Vaccination rates were also greater among those with comorbidities and risks.

Conclusions

By focusing on the TT resident's health events, we improved health modelization

Key messages

By using all the COVID information available in the database, we were able to show both, the beneficial effect of vaccination and the nefarious effect of COVID in a same model with machine learning. Aggregating hard categorical and numerical data with soft textual information about the resident’s health events, dates included, gives a precise and complete resident’s profile and health trajectory.