bluuebunny/crossref_metadata_2025_split huggingface dataset:
Significant participation bias and suboptimal cardiovascular risk management in learning healthcare systems due to non-consent
Authors: Zondag A G M , Groenhof T K J , Van Der Graaf R , Van Solinge W W , Bots M L , Haitjema S 
Published: November 2023

Abstract

Background
Computer-aided clinical decision support algorithms require continuous adjustments (learning) to account for changes in patient-characteristics, treatment(s), and outcomes, also referred to as a "Learning Healthcare System" (LHS). A LHS’s accuracy depends on adequate reflection of all patients, and participation bias may distort its effectiveness substantially. Therefore, we studied participation bias in a single center cardiovascular cohort study.

Methods
The cardiovascular cohort was set up as a LHS aiming for guideline-based treatment according to the individual risk profile for all cardiovascular disease patients in routine clinical care. Patients visiting an outpatient clinic for first time evaluation of a cardiovascular disease or risk factor received information on the cardiovascular cohort prior to their visit. During visit, cardiovascular risk management (CVRM) data was collected in the electronic health record (EHR) and informed consent was asked. We studied differences in characteristics between consenting, non-consenting and non-responding patients, and used multivariable logistic regression to identify determinants of non-consent. Multinomial regression was used for exploratory analyses of non-response determinants. A waiver (19/641) was obtained from the institutional review board for this study.

Results
In total, 2378 patients were consenting, 1907 non-consenting, and 1445 non-responding. In short, non-consent was related to higher cardiovascular risk (e.g., a cardiovascular disease history (OR 1.43, 95%CI 1.23-1.66)) and lower education (OR 0.76, 95%CI 0.60-0.97) than consent (Figure 1). Non-response, in contrast, was associated with higher physical activity (OR 1.04, 95%CI 1.01-1.08) and education level (OR 1.43, 95%CI 1.04-1.98) compared to consent. Non-consent and non-response patients were, respectively, younger and older. Presence of CVRM indicators in the EHR was 5-30% lower in non-consenting and 37-69% lower in non-responding patients compared to consenting patients.

Conclusion
A traditional informed consent procedure leads to participation bias in a LHS. Furthermore, it affects structured registration of CVRM indicators in the EHR, which is detrimental for the LHS feedback loop, and potentially leads to suboptimal cardiovascular risk management in patients not willing to participate. This study underlines the importance of reassessing the need of a traditional informed consent procedure for the use of routine care data in learning healthcare systems.
Endothelial cell-expressed long non-coding RNAs in cardiac regeneration
Authors: Kremer V , Josefs T , Phelp P , Hilbold E , Baer C , Boon R A 
Published: November 2023

Abstract
Endothelial cells (ECs) are the most abundant non-myocyte cell types in the heart. Upon cardiac injury, revascularization can occur through proliferation and migration of ECs from existing blood vessels in the surrounding myocardium. In animal models, revascularization is crucial to stimulate cardiomyocyte survival and regeneration. In the human heart, establishing blood flow could improve outcomes of tissue engineering or cell based therapy to regenerate the heart after injury. In addition to establishing blood flow, EC also secrete factors in a paracrine manner which can act on neighbouring cardiomyocytes.
We investigate a role for non-coding RNA in the regulation of endothelial cell function and their potential role in neovascularization and cardiac regeneration. A large portion of the genome is transcribed into non-coding RNA, which does not encode protein. Many long non-coding RNAs (lncRNAs) have been shown to be involved in important regulatory processes such as genomic imprinting and chromatin modification. To identify lncRNAs which could be involved in cardiac regeneration, we isolated ECs from neonatal mice at postnatal day 1 and 5.5. The potential for cardiac regeneration was observed in mice at postnatal day 1, but this capacity is lost by day 5.5. Differential expression of lncRNAs in ECs was assessed by RNA sequencing. Three potential target lncRNAs, which also have a human homologue, were selected. NR2F2-AS1 and HAND2-AS1 were upregulated at postnatal day 1, whereas FOXD2-AS1 was upregulated at day 5.5. These genes were selected for further validation in human cardiac microvascular endothelial cells (CMEC). Knockdown of NR2F2-AS1 resulted in a decrease in proliferation but did not affect angiogenic sprouting. Loss of HAND2-AS1 resulted in a trend to impaired sprouting but did not affect proliferation. FOXD2-AS1 knockdown resulted in a significant decrease in angiogenic sprouting. We observed no change in the rate of migration after knockdown of any of the three genes. Knockdown of the lncRNAs did not affect expression of the antisense protein coding transcripts, therefore the effects we observe are likely mediated by the lncRNAs.
In conclusion, lncRNAs NR2F2-AS1, HAND2-AS1 and FOXD2-AS1 regulate different aspects of EC function such as proliferation and sprouting. Modulation of expression of these transcripts therefore may be considered as a therapeutic tool to promote cardiac regeneration.
Regulation of circular RNAs in patients with cardiogenic shock
Authors: Baum L , Kokot K , Erbe S , Kneuer J , Buettner P , Thiele H , Laufs U , Boeckel J N 
Published: November 2023

Abstract

Background
The mortality rate of cardiogenic shock (CS) is around 50%. There is a need for biomarkers to predict the prognosis of CS. Due to their shape, circular RNAs (circRNAs) exhibit higher stability than other classes of RNA. Regulation of circRNAs has been observed in cardiovascular diseases such as myocardial infarction, but their regulation in CS is largely unknown. Monocytes can be classified into different populations based on their surface markers. Non-classical and intermediate monocytes are characterized by CD16 expression and have been associated with non-inflammatory function such as vascular healing.

Methods and Results
Blood samples were taken from patients with CS after myocardial infarction (MI) (n=4) on admission and 24 hours after revascularization and from healthy controls. Bioinformatic analysis by RNA-Sequencing revealed the expression of a total of 1819 circRNAs in the blood. 457 circRNAs showed altered expression in CS at the time of admission compared to controls, with 35 circRNAs upregulated and 422 circRNAs downregulated. Of these 457 circRNAs, 80 circRNAs were no longer regulated 24 hours after revascularization. This indicates dynamic regulation of circRNA during CS. Using divergently oriented primers spanning the backsplice junction of selected circRNA candidates, the presence of 17 of the most highly regulated or expressed circRNAs was confirmed by RT-PCR in peripheral blood mononuclear cells. We confirmed the circularity of the 17 circRNA candidates by RNAse-R digestion and oligo-dT selection. For one of the top candidates, circRNA-4, we confirmed upregulation in a larger group of patients with CS after MI (n=52) compared to healthy controls (n=38, 10.9-fold, p &lt; 0.0001). Interestingly, patients with MI without CS (n=37) showed no increase in circRNA-4 compared to healthy controls (n=31, p=0.2), suggesting a potential CS-specific regulation of circRNA-4 independent of myocardial infarction. Single cell sequencing of CS patients (N=4) revealed an increase in CD16+ monocytes and enriched expression of the circRNA-4 host gene. Interestingly, knockdown of circRNA-4 in human monocytes using siRNA targeting the circRNA-4-specific backsplice site resulted in a reduction of the non-classical monocyte marker CD16, suggesting a role for circRNA-4 in monocyte subpopulation switching.

Conclusion
In conclusion, the data reveal a dynamic and significant regulation of circRNAs in the blood of patients with CS after MI. circRNA-4 showed a CS-specific upregulation that was not primarily dependent on MI. After knockdown of circRNA-4, a reduced expression of the surface marker CD16 was also shown in monocytes indicating the involvement in monocyte subpopulation switch toward non-classical monocytes.
Thus, these data set the stage to further study circRNAs as potential new specific markers in acute diseases such as cardiogenic shock.
A better understanding of coronary artery disease molecular biology through an intermediate phenotype
Authors: Sa D , Mendonca M I , Santos M , Temtem M , Sousa F , Freitas D , Rodrigues M , Henriques E , Borges S , Guerra G , Ornelas I , Drumond A , Sousa A C , Palma Dos Reis R 
Published: November 2023

Abstract

Coronary Artery Disease (CAD) remains a common cause of death worldwide; over half of these deaths are asymptomatic until the first fatal presentation. Previous studies have often compared coronary artery calcification (CAC) with coronary stenosis or its sequelae. However, the genetic contribution to CAC in sub-clinical atherosclerosis is controversial.

Purpose
This study intended to assess the relationship between a set of single nucleotide polymorphisms associated with CAD (GWAS) and CAC score in an asymptomatic population.

Methods
Prospective study performed in an asymptomatic cohort from GENEMACOR population-based sample of 1207 subjects aged 51.7±8.3, 73.8 male, without apparent prior CAD. CAC score was performed by cardiac computed tomography and reported as Agatston units according to the Hoff nomogram (Low, Moderate and High-risk categories). For the present work, we considered two groups: Group 1 (0≤CAC&lt;100 and Percentile&lt;50) and Group 2 (CAC≥100 or Percentile≥50). We genotyped thirty-three single nucleotide polymorphisms (SNP) associated with CAD by TaqMan real-time PCR. Anthropometric, conventional, and biochemical risk factors were assessed. The association of these SNPs with CAC score groups were evaluated by bivariate and multivariate logistic regression analysis, and the dominant genetic model was considered for comparison.

Results
After bivariate analysis, only PHACTR1 rs1332844 C&gt;T (CT+TT genetic model) showed a significative association with CAC score (OR=1.45; 95%CI 1.09-1.94; p=0.011). Multivariate logistic regression analysis, adjusted to traditional risk factors, genetic model and CAC score showed that PHACTR1 rs1332844 remained in the equation as significantly associated to CAC score (P=0.009) together with age (0.0001), hypertension (0.006), Diabetes (0.0001), smoking (0.001) and obesity (0.028).

Conclusion
PHACTR1 genetic variant is known to contribute to atherosclerosis and plaque formation. In the present study it showed a significant association with plaque calcium. More research in this field is critical to understanding the genetic basis of CAD through an intermediate phenotype, plaque calcification. We highlight this point since it can be crucial for the prognosis and therapy of the asymptomatic population.
Elevation of CISD2 attenuates atrial aging
Authors: Yeh C H , Shen Z Q , Chou Y J , Seah C , Kao C H , Tsai T F 
Published: November 2023

Abstract

Background
Age-related atrial cardiomyopathy is characterized by conduction disturbance, structural fibrosis as well as cardiac remodeling. Atrial cardiomyopathy promotes the development of atrial fibrillation, which is associated with a 5-fold increase in the risk of stroke. However, the molecular mechanisms underlying age-associated deterioration of atrial function remain unclear. Cisd2 is a mitochondrial outer membrane protein that regulates intracellular calcium homeostasis and mitochondrial integrity. Importantly, Cisd2 is a pro-longevity gene in mammals.

Purpose
In this study, we investigate the role of Cisd2 in atrial cardiomyopathy during aging.

Methods
CISD2 expression and atrial conductance were examined in different age human surgical samples. Atrial conductance was assessed and validated by immunofluorescence and Transmission electron microscope in Cisd2 knockout and overexpressing transgenic mice at young and old age. The mitochondrial functions and calcium homeostasis were investigated in the atrial HL-1 cell lines with Cisd2KO or re-expression. Transcriptomic analysis of mice atrial tissues was performed.

Results
The age-associated decrease of atrial Cisd2 protein, in human and mice, were associated with atrial conductance disturbance and intercalated disc disorganization. Furthermore, Cisd2 deficiency disrupts calcium homeostasis via dysregulation of Serca2a and Stim1 resulting in an increased level of basal cytosolic Ca2+ and mitochondrial Ca2+ overload in HL-1 atrial cardiomyocytes. Most strikingly, in Cisd2 transgenic mice, a persistently high level of Cisd2 is sufficient to delay age-associated atrial myopathy and attenuate age-related atrial structural defects and functional decline. In addition, it results in a younger atrial transcriptome pattern during old age.

Conclusions
The mitochondrial-associated membrane-bounded CDGSH iron-sulfur domain-containing protein 2 (CISD2) protein regulates cytosolic calcium homeostasis and mitochondrial integrity. Interestingly, an age-dependent decrease of atrial Cisd2 protein has been detected during aging in both humans and mice. Our findings indicate that decreased atrial Cisd2 is one of the causes of atrial myopathy during cardiac aging. A persistently high level of atrial Cisd2 delays atrial myopathy during aging and ameliorates age-related atrial dysfunction.
Circulating large Extracellular Vesicles (EV-L) from STEMI patients contribute to post-injury adverse myocardial remodeling
Authors: Balbi C , Senesi G , Ministrini S , Brucale M , Valle F , Frigerio R , Bergese P , Cretich M , Akhmedov A , Camici G G , Vassalli G 
Published: November 2023

Abstract

Background
While primary percutaneous coronary intervention (PCI) is the treatment of choice in patients with ST-segment elevation myocardial infarction (STEMI), this treatment does not fully prevent late adverse cardiac remodeling. Infarcted myocardial tissue releases damage-associated extracellular vesicles (EV) that mediate tissue inflammation and may regulate myocardial remodeling. Here, we investigated the role of circulating EV in STEMI patients, with a focus on endothelial cell (EC) injury and myocardial fibrosis.

Methods
Peripheral blood samples were obtained from STEMI patients (n=30) within 4 hrs of onset of pain before PCI and from age- and gender-matched healthy control (CTRL; n=30) with documented absence of coronary artery disease. Raw serum was characterized by atomic force microscopy (AFM) single-particle morphometry. Large EV (EV-L) were isolated by differential centrifugation and used for in vitro experiments on human aortic endothelial cells (hAEC), and human cardiac fibroblast (hCF).

Results
AFM analysis showed a 7.5-fold increase in EV-L in STEMI vs. CTRL serum. In vitro treatment of hAEC with EV-L from STEMI patients, but not CTRL-EV-L, induced an increase in reactive oxygen species (ROS; 1.15-Fold increase), p66SHC and oxidized low-density lipoprotein receptor 1 (LOX-1) expression (1.34 and 20-Fold increase, respectively), and cell death (1.40-Fold increase). Interestingly, ELISA analysis on EV-L showed an enrichment of LOX-1 in STEMI-EV-L compared to CTRL-EV-L (3.01-Fold increase). Moreover, STEMI-EV-L, but not CTRL-EV-L, stimulated in vitro proliferation (1.4-Fold increase) of hCF and their activation in myofibroblasts, as assessed by alpha smooth muscle actin (αSMA) expression (1.8-fold increase), resulting in increased extracellular matrix (ECM) secretion (1.5-Fold increase).

Conclusions
Our preliminary data showed a pathological role of circulating EV-L derived from STEMI patients. In particular, results from in vitro experiments evidence their function in inducing endothelial damage and activation of cardiac fibroblast into myofibroblast; two important features in the progression of pathological myocardial remodeling.Graphical abstract
Serpina3c attenuates adipose tissue and heart damage during obesity by inhibiting oxidative stress and endoplasmic reticulum stress
Authors: Jiang Y , Guo J Q , Wu Y , Yao Y Y 
Published: November 2023

Abstract

Background
Adipose tissue (AT) dysfunction is closely associated with obesity-related heart damage. Serpina3c which is highly expressed in mature adipocytes, is a secreted serine protease inhibitor that can regulate AT function. The present study aimed to investigate the effect of Serpina3c on AT and heart during obesity.

Methods
Wild type (WT) and Serpina3c knockout (3cKO) mice were fed with high-fat diet (HFD) for 20 weeks. AAV-mediated overexpression of Serpina3c was injected locally in epididymal white adipose tissue (eWAT) to examine the effect of Serpina3c. Palmitic acid was established to interfere with the differentiated mature 3T3L1 cells of Serpina3c knockdown (3cKD) or overexpression (3cOV) and corresponding control groups. RNA-seq of mature 3T3L1 cells was performed to assess the effect of Serpina3c on the transcriptome.

Results
Decrease of Serpina3c protein in AT of obese mice induced by HFD. The body weight and serum cholesterol, triglycerides, free fatty acid and pro-inflammatory cytokines were increased significantly in 3cKO mice. Increased inflammation, fibrosis and apoptosis of AT as well as cardiac fat deposition, inflammation and apoptosis were detected, and the impairment of cardiac diastolic function was aggravated in KO mice. Overexpression of Serpina3c in eWAT alleviated these adverse phenotypes. Through RNA-seq, we found endoplasmic reticulum stress (ERS)-related genes were significantly increased in 3cKD group, but significantly down-regulated in 3cOV group. Oxidative stress was generally considered to be the upstream of ERS, and we detected a significant increase in ROS levels in 3cKO group. Serpina3c alleviated ERS and production of pro-inflammatory cytokines in adipocytes by inhibiting NOX4 and iNOS mediated oxidative stress.

Conclusion
Our results highlight a protective role for Serpina3c as a novel adipocytokine in the obesity-related heart damage, with Serpina3c conferring protection through inhibiting NOX4 and iNOS mediated oxidative stress to negatively regulate ERS and inflammation in adipocytes.Schematic illustration
Estimating the true half-life of cardiac troponins, a novel methodological approach
Authors: Kristensen J H , Hasselbalch R B , Strandkjaer N , Bundgaard H , Iversen K K 
Published: November 2023

Abstract

Introduction
Cardiac troponins (cTn) are key when diagnosing myocardial injury (MI) and are used worldwide. Still, the half-life (T½) of cTn in the bloodstream is largely unknown. Previously, T½ has been estimated to 7-20 hours following MI. However, in the clinical setting the true T½, i.e. the decay of cTn, cannot be estimated due to ongoing leakage of cTn from cardiomyocytes. Identification of the true T½ is essential to improve estimation of how long cTn leak from cardiomyocytes due to MI and to understand the pathophysiology of all diseases in the myocardium associated with raised cTn levels.

Purpose
To determine T½ of cTnI and cTnT in the human blood stream.

Methods
Patients with ST elevation myocardial infarction (STEMI) were included within 24 hours after acute reperfusion therapy and underwent plasmapheresis to "harvest" plasma with a high cTn concentration. Three to 17 weeks later patients received a re-transfusion of autologous plasma. Blood samples were drawn at 22 fixed time points over eight hours from re-transfusion. A two phased model was used to estimate T½ of cTnI and cTnT for all patients. An alpha level of &lt;0.05 was considered statistically significant and differences were tested by Wilcoxon matched pairs signed rank test.

Results
A total of 20 participants received a re-transfusion of their own plasma. Median age was 63 years (IQR 58-70), and four participants were women (20%). Prior to re-transfusion the participants’ median concentration of cTnI was 16 ng/l (interquartile range (IQR) 12-24) and of cTnT 12 ng/l (IQR 11-18). Following re-transfusion of 180-679 ml autologous plasma, participants’ concentration of cTn was measured again. Median concentration of cTnI was 3,355 ng/l (IQR 2,135-4,690) and of cTnT 237 ng/l (IQR 160-303 ng/l). Decay of cTnI and cTnT from injection to eight hours after followed a two-phased exponential decline with a primary fast and secondary slow phase as presented in figures 1-2. Each colored curve represents a participant. Median T½ during the first phase was 11 min for cTnI (IQR 6-13 min) and 15 min for cTnT (IQR 7-18 min). The fast decline in the first phase was thought to be due to re-distribution in the extracellular space. The second phase had a median T½ of 65 min for cTnI (IQR 56-80 min) and 67 min for cTnT (IQR 51-92 min). The second phase represents the human, biological T½ of cTn. No significant difference was found between T½ of cTnI and cTnT.

Conclusions
This novel study design showed seven to 20-fold shorter T½ of cTnI and cTnT compared to previous estimates in the acute, clinical setting, indicating that the duration of cTn leakage from the myocardium during MI has previously been grossly underestimated.
This finding challenges our understanding and interpretation of the clinical course of MI and may translate into changes in the perceived window of opportunity for myocardial salvage by interventions.Figure 1:Cardiac troponin IFigure 2:Cardiac troponin T
Molecular alterations of human diabetic heart disease revealed using single nucleus RNA sequencing
Authors: Gollmer J , Horstmann H , Potter L , Vosko I , Tomin T , Pfeil K , Briner-Gruenberger R , Von Lewinski D , Sedej S , Scherr D , Wende A R , Rainer P , Zirlik A , Wolf D , Bugger H 
Published: November 2023

Abstract
Type 2 diabetes mellitus (T2D) increases the risk for heart failure. Whether mechanisms proposed to underlie cardiac structural and functional alterations in the rodent diabetic heart are recapitulated in humans remains poorly investigated. Here, we studied left ventricular samples of 8 subjects with T2D, preserved ejection fraction (63±5%) and no history of ischemic heart disease (Db-pEF), 7 subjects with T2D, reduced ejection fraction (26±9%) and ischemic heart disease (Db-ICM), and 15 non-diabetic individuals with normal ejection fraction (Non-Db-pEF) serving as controls. 1168 proteins were identified using label-free proteomics by LC-MS/MS. 146 were differentially expressed in Db-ICM, but only 66 in Db-pEF. Bulk RNA sequencing revealed differential expression of 1795 and 527 genes in Db-ICM or Db-pEF, respectively, with only 128 genes being commonly regulated. Pathway analysis revealed a significant regulation of pathways related to inflammation or extracellular matrix remodelling in Db-ICM and Db-pEF, however literally no enrichment in pathways previously related to diabetic cardiomyopathy. To characterize the cardiac cellular heterogeneity, we performed single nucleus RNA sequencing (snRNAseq) that showed no differences in cell frequencies between the groups. However, analysis of genes differentially expressed in cardiomyocytes of diabetic hearts unmasked enrichments in many pathways related to rodent diabetic cardiomyopathy not observed by proteomics or RNA sequencing, including insulin resistance, fatty acid oxidation, oxidative phosphorylation, oxidative stress, and various signaling pathways. Furthermore, interactome analysis revealed fewer cell-cell interactions in Db-pEF and Db-ICM compared to controls and implied differential activation of several signaling pathways to contribute to diabetic heart disease, including adiponectin, hepatocyte growth factor, and natural cell adhesion molecule signaling. Thus, we present the first comprehensive atlas of molecular and cellular alterations in the human diabetic heart, revealing recapitulation of many pathomechanisms previously proposed in animal studies. This analysis represents a decisive step towards understanding the pathology underlying human diabetic heart disease and may help develop future diagnostic and therapeutic strategies.