bluuebunny/crossref_metadata_2025 huggingface dataset:
Understanding of seasonal influenza immunisation and its cardiovascular benefits among cardiology patients
Authors: Monagle S R , Spear E , Thakur U , Pol D , Tan S , Bellamy K , Hickman J , Jackson B , Chan J , Nicholls S J , Nelson A J
Published: November 2023

Background

Seasonal influenza immunisation reduces the risk of cardiovascular events. Patients with established cardiovascular disease (CVD) derive a greater benefit than those without yet up to 50% globally do not take up the immunisation.

Purpose

To describe the understanding and perceptions of seasonal influenza immuni2ation among patients CVD.

Methods

A cross sectional, voluntary and anonymous survey was administered to cardiology inpatients and outpatients attending a large health system in Australia during the 2022 influenza season, May 1st 2022 to August 31st 2022. Demographics, educational and employment status and prior immunisation behaviours were collected. Questions on perceptions and beliefs around seasonal influenza immunisation and influenza-related illness were presented with 5-point Likert scale (strongly disagree through to strongly agree).

Results

A total of 181 surveys were completed; median age was 64 (50.5% <65 years), 65.0% (115/177) of the population were inpatients, 82.2% (143/174) had completed high school or more; 35.0% (63/180) were female and 24.2% (43/178) spoke a language other than English at home. Over one third of respondents did not receive the seasonal influenza immunisation in the prior year (63/181, 34.5%). Only half (97/179, 54.2%) of patients agreed that their heart condition placed them at higher risk of complications and serious illness if they contracted influenza. While 73.3% (132/180) agreed that immunisation would help prevent risk of serious illness, only 26.8% (48/179) agreed that it would help to prevent myocardial infarction. Nearly a quarter of patients (24.0%, 43/179) were concerned about side effects while 1 in 10 (12.8%, 23/179) patients raised cost as a barrier despite being free-of-charge to all patients with CVD in Australia. If asked to receive the seasonal influenza immunisation, 86% (154/179) patients would agree if their cardiologist recommended it.

Conclusions

Despite guideline recommendations, most cardiology patients are uninformed of the cardiovascular benefits of seasonal influenza immunisation with many unaware they are at higher risk of influenza-related, preventable illness. The vast majority of patients would accept the immunisation if recommended by their cardiologist highlighting their important role in improving uptake among this high-risk group.
Level of evidence supporting the chinese cardiovascular disease clinical practice guidelines and its evolution in the past two decades
Authors: Li C , Wang C S , Hao J , Zheng Y T , Yang J , Tang Y D
Published: November 2023

Background

Clinical practice is guided by guidelines in the era of evidence-based medicine to improve healthcare. The consistency between the strength of recommendations and the underlying quality of evidence in clinical guidelines and its evolution dynamically reflects the status of medical practice in important aspects. This study aimed to evaluate the levels of evidence (LOEs) supporting different classes of recommendations (CORs) in Chinese cardiovascular disease (CVD) guidelines between 2003 and 2021 and changes over time.

Method

Clinical guideline documents on cardiovascular topics issued by leading professional organizations were retrieved in the Databases of SinoMed and Wanfang Med Online from inception to June 2021. All guidelines were screened through abstract and full-text reading and included if satisfying the pre-specified criteria. 79 Chinese guideline documents on 12 sub-topics including a total of 5195 recommendations and the designated CORs/LOEs, were abstracted. The number of recommendations of Class I, Class II, Class III, LOE A, LOE B and LOE C were identified for each guideline document. The proportion of CORs, LOEs and COR-LOE combinations in guidelines and the changes among those with ≥2 versions.

Results

A total of 79 guidelines were included in the analysis. When examining the status of current guidelines, among the 3325 recommendations derived from 59 documents during 2011-2021, 735 recommendations (22.1%) were classified as LOE A, 1280 (38.5%) as LOE B, and 1310 (39.4%) as LOE C. 596 recommendations (17.9%) were characterized as Class I-LOE A, accounting for the majority of LOE A recommendations but only one-third of Class I recommendations. Evidence levels varied greatly across different sub-topics and individual guidelines. There are 9 guidelines on 5 sub-topics having ≥2 versions. When analyzing the changes over time, although an increase was observed in the total number of recommendations, the proportion of recommendations designated as Class I-LOE A did not significantly improve (19.1% [current] vs 19.0%[prior], p=0.97).

Conclusions

In current Chinese CVD guidelines, the high level of evidence lacks and its alignment with strong recommendations is deficient. Although it shows moderate improvements in certain major topics (e.g., Coronary artery disease, Interventional therapy, Surgery) in the past two decades, the overall proportion of Class I-LOE A recommendations remains small, suggesting that conduction, and particularly translation, of high-quality studies like RCTs addressing CVDs-related questions are still essential and demanded, especially for areas with less attention.Abstract Picture 1
Venous thromboembolism in users of hormonal contraception and non-steroidal anti-inflammatory drugs: a nationwide cohort study
Authors: Meaidi A , Mascolo A M , Sessa M S , Toft-Petersen A P T P , Skals R S , Gerds T A G , Skovlund C W S , Morch L M , Rossi F R , Capuano A C , Lidegaard O L , Torp-Pedersen C T P
Published: November 2023

Objective

To study the influence of concomitant use of hormonal contraception and non-steroidal anti-inflammatory drugs (NSAIDs) on the risk of venous thromboembolism.

Design: Nationwide cohort study.

Setting: Denmark through national registries.

Participants: All 15-49-year-old women living in Denmark between the years 1996-2017 with no medical history of any venous or arterial thrombotic event, cancer, thrombophilia, hysterectomy, bilateral oophorectomy, sterilisation, or infertility treatment.

Main outcome measure: A first-time discharge diagnosis of lower-limb deep venous thrombosis or pulmonary embolism.

Results

Among 2.0 million women followed for 21.0 million person-years, 8710 venous thromboembolic events occurred. Compared to non-use of NSAIDs, use of NSAIDs was associated with an adjusted incidence rate ratio of venous thromboembolism of 8.1 (95% confidence interval (CI) 6.9 to 9.6) in women not using hormonal contraception, 12.1 (10.6 to 13.9) in women using high-risk hormonal contraception, 8.7 (6.5 to 11.6) in users of medium-risk hormonal contraception, and 5.1 (2.9 to 9.1) with low/no-risk hormonal contraception.

Compared to non-use of NSAIDs, the number of extra venous thromboembolic events per 100,000 one-week NSAID treatments was 4 in women not using hormonal contraception, 18 in women using high-risk hormonal contraception, 9 in users of medium-risk hormonal contraception, and 2 in women using low/no-risk hormonal contraception.

Conclusions

NSAID use was positively associated with the development of venous thromboembolism. The association was stronger in women concomitantly using high-risk hormonal contraceptives and weaker in women using low/no-risk hormonal contraception. Women requiring both hormonal contraception and regular use of NSAIDs should be advised accordingly.
The impact of chronic kidney disease on healthcare utilization and costs associated with pacemaker infection
Authors: Wright D J , Trucco M E , Marti M A , Margetta J , Wolff C , El-Chami M F
Published: November 2023

Background

Patients with CKD are at increased risk of bleeding and device-related infections due to their chronic illness. CKD complicates treatment which could lead to an increase in duration of hospitalizations and higher costs.

Purpose

The goal of this study is to compare hospitalizations and the total costs associated with permanent pacemaker (PPM) infection in patients with and without CKD.

Methods

This retrospective analysis used Medicare Fee for service (FFS) claims to identify patients implanted with a transvenous PPM from June 2016-December 2020. Device infections, hospitalizations and costs were identified in a 12-month follow-up period post-device implant. Costs include all inpatient and outpatient hospital costs. Patients with CKD and infections were identified based on ICD-10 diagnostic codes. A generalized linear model was utilized to calculate non-infection and infection costs by CKD status. The simple differences between these cohorts represent the total infection cost with CKD and total infection cost without CKD highlighting the incremental cost of infected patients with CKD.

Results

A total of 439,679 Medicare FFS patients had PPM insertion from June 2016-December 2020, of which 13% had CKD and 1.2% had a device infection within 12-months post-implant. Infected patients with CKD incurred two admissions on average (p-value = <0.001), versus 1.4 (p-value = <0.001) for an infected patient without CKD. The total days in hospital for infected patients with CKD were 32 days (p-value = <0.001) versus 22 days (p-value=<0.001) for infected patients without CKD (Figure 1). The average cost of an infection in the US for a CKD patient was $176,483 (p-value = <0.001) versus $65,792 (p-value =<0.001) for a patient without CKD. The incremental costs of an infection in CKD as compared to no CKD was $110,691 (p-value of interaction = 0.0160).

Conclusion

In our analysis PPM-related infection in patients with CKD were characterized by more health care utilizations and higher costs compared to patients without CKD.Figure 1
Bystander defibrillation increases 30-day survival even with short emergency medical service response time
Authors: Hindborg M , Yonis H , Gnesin F , Soerensen K K , Torp-Pedersen C
Published: November 2023

Background

Rates of bystander initiated defibrillation are increasing in many areas of the world (1-4), but the effect of defibrillation at different time points after recognition of out-of-hospital cardiac arrest (OHCA) remains largely unknown.

Purpose

To assess the effect on 30-day survival of bystander defibrillation at different intervals of emergency medical service (EMS) response time.

Methods

We included OHCAs from 2016 through 2020 from the Danish Cardiac Arrest Registry. Cases were included if they were ≥18 years of age, bystander witnessed, received bystander cardiopulmonary resuscitation (CPR), had EMS response time of 25 minutes or less and only the first OHCA for each subject was considered. We excluded cases witnessed by EMS or if they had missing values of OHCA-related variables.

Crude survival proportions were calculated for each minute of EMS response time.

Relative risks (RR) and corresponding 95 % confidence intervals (95 % CI) of the outcome (30-day survival) for eight different intervals of EMS response time were estimated using a causal inference framework utilizing Targeted Maximum Likelihood Estimation (TMLE) with ensemble Super Learning, adjusting for age, sex, place of arrest (public/private), initial cardiac rhythm (shockable/not shockable), and comorbidities (prior acute myocardial infarction, ischemic heart disease, heart failure, chronic obstructive pulmonary disease, stroke, cancer).

Results

We included 7,471 cases of bystander witnessed OHCA receiving CPR before EMS arrival. Of these, 14.7 % (1,098/7,471) received bystander defibrillation before arrival of EMS. Overall, 44.5 % (489/1,098) survived to 30 days when bystander defibrillation was performed versus 18.8 % (1,200/6,373) when no bystander defibrillation was performed. When examining the crude survival proportion for each minute of EMS response time, we found that 30-day survival was consistently higher in the group receiving bystander defibrillation for the first 20 minutes after which the survival was approximately the same for the two groups (Figure 1). When adjusting for confounders, we found statistically significantly increased relative risks of 30-day survival for patients receiving bystander defibrillation, compared to patients not bystander defibrillated, for all intervals of EMS response time, except for response times of 0-2 min., where the increase did not reach statistical significance (0-2 min: RR; 1.34 [95 % CI: 0.88-2.05], 2-4 min: RR; 1.37 [95 % CI: 1.10-1.70], 4-6 min: RR; 1.55 [95 % CI: 1.33-1.80], 6-8 min: RR; 2.23 [95 % CI: 1.86-2.67], 8-10 min: RR; 1.99 [95 % CI: 1.55-2.55], 10-12 min: RR; 1.89 [95 % CI: 1.34-2.68], 12-15 min: RR; 1.86 [95 CI: 1.22-2.84], >15 min: RR; 1.98 [95 % CI: 1.16-3.38]) (Figure 2).

Conclusion

For all intervals of EMS response time, bystander defibrillation increased 30-day survival of OHCA. The effect was significant for EMS response times as short as 2-4 minutes.
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

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

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

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 < 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

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<100 and Percentile<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>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

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

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

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

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 <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

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.
Dysbiotic microbiome induce cardiac impairment by attenuating brain-derived neurotrophic factor in inflammatory bowel diseases
Authors: Li Q , Zhong X S , Krishnachaitanya S S , Liu M , Ou R , Xiao Y , Powell D W , Fujise K
Published: November 2023

Background & Aims

Mounting evidence suggests that inflammatory bowel disease (IBD) is associated with increased incidences of cardiovascular diseases (CVD), including myocardial infarction and heart failure. However, the molecular mechanisms by which IBD predisposes patients to CVD are poorly understood. This study aimed to identify the role of dysbiosis in gut-heart crosstalk.

Methods

Chronic colitis-induced cardiac impairment was assessed in mice subjected to dextran sodium sulfate (DSS), rats treated with trinitrobenzenesulfonic acid, and IL10-/-mice which develop spontaneous colitis after weaning. Fecal microbiota of wild type or IL10-/- mice were transplanted to C57BL/6J mice treated with microbiome-depleting antibiotic cocktail and germ-free mice to determine if dysbiosis afflicts the heart. Heart function was evaluated by echocardiography. Heart transcriptomes were profiled by RNA-sequencing. Gut microbiota profiling was performed by 16S rRNA sequencing. Adverse effect of brain-derived neutrophic factor (BDNF) deficiency on the heart was evaluated in cardiac-specific BDNF knockout mice (BdnfΔheart).

Results

Impairment of heart function, evidenced by decreased left ventricle ejection fraction (LVEF), increased LV mass, and elevated serum B-type natriuretic peptide, was observed in all 3 models of colitis, providing direct evidence that chronic colitis deteriorates the heart (Figure 1). RNA-sequencing revealed loss of Bdnf and alterations of multiple genes critical to heart homeostasis in the myocardium. Many of the changes, including Bdnf reduction, were found in microbiota-depleted C57BL/6J mice and germ-free mice received dysbiotic fecal microbiome, compared to controls, suggesting that dysbiosis impaired the heart by suppressing Bdnf. BdnfΔheart mice had impaired heart function, including decreased contractility and EF, increased fibrosis, and impaired coronary microcirculation (lower CD31), compared to Bdnf[2lox] mice. Mechanistically, miR-155 in the heart and exosomal miR-29b in the plasma were significantly elevated by chronic colitis. miR-155 and miR-29b mimics suppressed BDNF in cardiomyocytes both in vitro and in vivo. Administration of miR-155 inhibitors or exosomes packaged with miR-29b antagonists ameliorated BDNF reduction in the heart of IL10-/- mice. BDNF supplementation markedly improved heart function in DSS mice.

Conclusions

Chronic colitis impairs the heart through dysbiosis/miR-29b/miR-155/BDNF signaling axis. Targeting this pathway is a promising strategy to overcome gut dysbiosis harmful for the heart in IBD.
Identification of cell-intrinsic effects of mutated circulating cells due to clonal hematopoiesis in patients with heart failure
Authors: Abplanalp W , Schuhmacher B , Cremer S , Merten M , Macinkovic I , Shumliakivska M , Zeiher A M , John D , Dimmeler S
Published: November 2023

Clonal hematopoiesis of indeterminate potential (CHIP) is caused by somatic mutations in hematopoietic stem cells and is associated with worse prognosis in patients with heart failure with reduced ejection fraction (HFrEF). Circulating immune cells derived from patients with HFrEF harboring CHIP mutations show enhanced proinflammatory signatures. However, it is unclear, whether these signatures are derived from the relatively low number of cells harboring CHIP-driver mutations, or are indicators of a systemic pro-inflammatory activation associated with CHIP.

Methods and Results

We assessed the cell-intrinsic effects of the most prevalent CHIP-driver gene (DNMT3A) mutant-carrying circulating immune cells in a total of 5 patients with HFrEF. Using an improved protocol to detect mutant cells on a single cell level (MutDetect-Seq), as illustrated in figure 1A, we show that CHIP mutations can be detected in all major immune cell types (e.g. T cells, monocytes, B cells, NK cells). DNMT3A mutant monocytes demonstrated the highest number of significantly upregulated genes relative to other cell types. Specifically, DNMT3A mutant monocytes exhibit altered gene expression profiles associated with inflammation and phagolysosome function (figure 1B). In addition, DNMT3A mutant carrying CD4+, but not CD8+ T cells, showed significant changes in gene expression patterns towards effector cells and away from a naïve T cell state. Finally, increased paracrine signaling pathways involved in amplifying inflammatory circuits are predicted between mutated and non-mutated monocytes.

Conclusion

This is the first study to decipher the genetic signature of DNMT3A-CHIP-driver mutation carrying circulating immune cells, thereby providing first evidence for the cell intrinsic effects of DNMT3A mutations on peripheral blood cell subsets in patients with HFrEF. The altered gene expression profiles associated with inflammation and phagolysosome function of mutant monocyte as well as the skewing of mutated T cells towards effector cells provide novel insights into how CHIP might promote worse prognosis in heart failure patients.MutDetect-seq
Hematopoietic loss of Y-chromosome in monocytes from heart failure patients associates with cardiac fibrosis signaling
Authors: Abplanalp W , Cremer S , John D , Merten M , Schuhmacher B , Macinkovic I , Dimmeler S , Zeiher A M
Published: November 2023

Background

Hematopoietic mosaic loss of Y chromosome (LOY) increases with age and associates with increased cardiovascular disease incidence. Furthermore, recent murine studies suggest that LOY could potentially promote cardiac fibrosis by stimulating the infiltration and activation of bone marrow-derived macrophages. Yet, specific signatures of hematopoietic LOY cells in patients with heart failure (HF) are missing.

Methods and results

To decode the genetic signature of circulating hematopoietic cells and assess the effects of LOY in human leukocytes, we performed single-cell RNA-sequencing (scRNA-seq) of immune cells from 10 male patients with chronic HF (mean age 67.3 years, ejection fraction 34.4%). We sorted immune cells by presence or absence of Y-chromosome-associated genes (figure 1A). Relative distribution of LOY was assessed, with monocytes having the greatest absolute count of LOY cells with no significant change in distribution among cell types. Given the abundance of monocytic LOY cells we investigated LOY monocytic cells transcriptional signatures (29810 Y-carrier monocytes, 2205 LOY monocytes) in an unbiased manner. Using a patient-specific, paired analysis, LOY monocytes showed significantly reduced expression of TGF-ß inhibiting genes (SMAD7, TGIF2) in LOY cells versus Y-carrier cells. Opposingly, from 193 upregulated genes, LOY cells showed elevated markers associated with monocyte/macrophage-mediated tissue damage and pro-fibrotic cardiac remodelling (S100A8, CLEC4D, TLR2). Gene ontology terms derived from upregulated genes in LOY cells further supported a mechanistic link to cardiac fibrosis by calling terms associated with TLR signaling, which are known to enhance cardiac fibrosis. (figure 1B).

Conclusion

This is the first study to decipher the genetic signature of LOY monocytes in HF patients. Our results considerably extend very recent experimental findings in mice that LOY sensitizes macrophages to enhanced profibrotic signaling. Diminished TGF-ß inhibitory molecules and enhanced TLR signaling may contribute to the aggravation of heart failure by LOY in patients with HF.HF mLOY scRNA-seq
Non-neuronal acetylcholine is causal for cardioprotection by hypoxic preconditioning in adult rat cardiomyocytes
Authors: Braczko F , Reinders J , Lieder H R , Heusch G , Kleinbongard P
Published: November 2023

Introduction

Brief episodes of non-lethal myocardial ischemia/reperfusion preceding more sustained ischemia and reperfusion reduce lethal ischemia/reperfusion injury. The underlying protective signaling mechanisms of such ischemic preconditioning are unclear in their details. Among other protective mediators, neuronal acetylcholine (ACh) and its signaling through cholinergic receptors is involved in protection by ischemic preconditioning. Recently, a non-neuronal cardiac cholinergic system (NNCCS) has been identified, suggesting that a nervous system-independent, cardiomyocyte-derived acetylcholine signaling with signal amplification by auto- and paracrine cholinergic receptor activation could also participate in ischemic preconditioning.

Aim

We aimed to examine the role of NNCCS-derived ACh as a mediator of ischemic preconditioning. We therefore, in a reductionist approach, used isolated adult cardiomyocytes with hypoxic preconditioning (HPC).

Methods

Adult ventricular rat cardiomyocytes were isolated (from n=9 hearts), and HPC was induced by one cycle of 10 min hypoxia / 20 min reoxygenation. Thirty min normoxia served as control. Cardiomyocytes were then exposed to 30 min sustained hypoxia / 5 min reoxygenation (H/R), or a 35 min time control (TC). Cardiomyocyte viability was quantified before and after H/R or TC, respectively, by trypan blue staining. In a subgroup of experiments (cardiomyocytes from n=6 hearts) in the presence of physostigmine (0.2 mmol/L), intracellular ACh was quantified before and after HPC or normoxia and after sustained hypoxia and reoxygenation or TC, respectively, via liquid chromatography-coupled tandem mass spectrometry. Experiments on cardiomyocyte viability (from n=9 hearts) were repeated with the addition of cholinergic receptor antagonists (nicotinic: hexamethonium [1 µmol/L], muscarinic: atropine [0.1 µmol/L]) a) during the first 30 min with HPC or normoxia, b) during H/R or TC (30-65 min), or c) throughout the total 65 min protocol.

Results

The viability of cardiomyocytes subjected to H/R was better preserved with HPC than without (Figure A), and intracellular ACh was increased during hypoxia after prior HPC (Figure B). Cholinergic receptor antagonists during HPC did not change HPC´s protection (Figure A). However, the combination of both antagonists during sustained hypoxia abrogated HPC´s protection (Figure A), whereas nicotinic or muscarinic cholinergic receptor antagonists alone did not. Cholinergic receptor antagonists throughout the total 65 min also abrogated HPC´s protection (Figure A). Conclusion: The NNCCS is activated by HPC and causal for HPC´s protection. HPC increases intracellular acetylcholine, and cardiomyocyte protection is mediated via both nicotinic and muscarinic cholinergic receptors, suggesting an auto- and paracrine signal amplification on the cardiomyocyte level.Figure