bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of Energy Consumption for electric motorcycles depending on vehicle configuration and driving cycles
Authors: Niccolai A , Berzi L , Barone F , Baldanzini N
Published: May 2024

This article presents an analysis of energy consumption for battery electric motorcycles (BEM) based on vehicle configuration and driving cycles. The study investigates the impact of two architectures: rear-wheel regenerative (RWR), and two-wheel regenerative (2WR) on consumption. For the analysis, the study considered the effect of various parameters on the longitudinal vehicle dynamics such as speed profiles (WMTC, real-world profile), longitudinal road slope (from -5° to 5° with 0.5° step increment), and 5 different brake distributions. The research employs simulation techniques using the Matlab/Simulink environment to develop a simplified longitudinal vehicle dynamics model (1 DOF) and for the implementation of a serial brake-blending control strategy. The results of the analysis provide energy potential regeneration and insight into the sensitivity of energy consumption to road slope variations, powertrain working points, and potential energy harvesting. The findings contribute to a valuation of the factors that influence energy consumption in electric motorcycles and have implications for the development of vehicle architectures through accurate range assessment on real-world riding conditions and provide valuable information for powertrain components right-sizing.
Impact of sand feed rate on the damage of railway wheel steels
Authors: Zani N , Shu K , Ghidini L , Petrogalli C , Mazzù A
Published: May 2024

The global railway industry plays a pivotal role in economic development, offering efficient transportation solutions. However, railways operating in desert environments face unique challenges due to windblown sand. This study investigates the influence of sand feed rates on wheel-rail wear in desert conditions. Experimental tests were conducted using a bidisc apparatus to simulate sand feed rates. Results indicate that low feed rates lead to spalling and pitting, while high rates increase abrasive and fatigue wear. A critical transition point at 0.4 g/min suggests sand-induced abrasion of wheel surfaces. Moreover, the research highlights the crucial role of sand feed rates not only in wear but also in surface roughness, further emphasizing the complex interplay between sand transport rates, adhesion, and wear mechanisms. These insights provide valuable guidance for mitigating wear-related challenges in desert railway operations and optimizing maintenance strategies.
Product Re-Design Towards Sustainability: a Case Study of a Mass-Production Device
Authors: Moreschini M. , Migliavacca A. , Mansutti A. , Ilare D. , Guagliano M.
Published: May 2024

Sustainability issues are nowadays embedded in society and organizations cannot avoid tackling them. Product design represents the phase in which 80% of the overall emissions are determined, thus the one on which changes should be realized. In some industrial sectors the environmental impact has never been accounted systematically during the conceptualization phase. The resulting products are consequently characterized by poor environmental performances, that have a deep negative effect if realized on a mass production scale. A sustainable re-design and process innovation approach is therefore required. This is the objective of this research, that uses Rold Washing Machine Doorlock as a case study example. After the realization of a cradle-to-gate Life Cycle Assessment (LCA) of the component under exam, design and process modifications are identified. The assessment is not limited to the proposal of environmentally improved versions of the product. A barrier towards sustainability adoption is represented by the common perception of its unaffordability. It is important for the identified solutions leading to improved ecological performances, but they should be sustainable for the company also in financial terms. At this purpose, for each proposed change, a technical assessment of the operational steps required is performed, together with an economic analysis. The resulting re-engineered product passes all the desired conditions, and the methodology applied can be extended to every other sector facing the same issue.
Development of a biofabricated 3D in vitro vessel model for investigating transendothelial migration in stem cell therapy
Authors: Wachendörfer Mattis , Palkowitz Alena Lisa , Fischer Horst
Published: May 2024

Systemic stem cell therapies hold promise for treating severe diseases, but their efficiency is hampered by limited migration of injected stem cells across vascular endothelium towards diseased tissues. Understanding transendothelial migration is crucial for improving therapy outcomes. We propose a novel 3D in vitro vessel model that aids to unravel these mechanisms and thereby facilitates stem cell therapy development. Our model simulates inflammation through cytokine diffusion from the tissue site into the vessel. It consists of a biofabricated vessel embedded in a fibrin hydrogel, mimicking arterial wall composition with smooth muscle cells and fibroblasts. The perfusable channel is lined with a functional endothelium which expresses vascular endothelial cadherin, provides an active barrier function, aligns with flow direction and is reconstructed by in situ two-photon-microscopy. Inflammatory cytokine release (tumor necrosis factor α, stromal-derived factor (1) is demonstrated in both a transwell assay and the 3D model. In proof-of-principle experiments, mesoangioblasts, known as a promising candidate for a stem cell therapy against muscular dystrophies, are injected into the vessel model, showing shear-resistant endothelial adhesion under capillary-like flow conditions. Our 3D in vitro model offers significant potential to study transendothelial migration mechanisms of stem cells, facilitating the development of improved stem cell therapies.
Influences of preheating parameters on the quality of weld by thermite rail welding
Authors: Burapa Romadorn , Oo Hein Zaw , Sangwiman Wissarut , Muangjunburee Prapas
Published: May 2024

The major goal of this study is to enhance the mechanical and metallurgical characteristics of rail steel grade R260 joined by thermite welding under various preheating conditions, including preheating time and gas pressure. Mainly two conditions, referred to as the Normal Condition and Improved Condition, are carried out for experiments. Prior to welding, the Normal Condition was preheated using liquefied petroleum gas (LPG) and oxygen gas pressures of 1 bar and 4.5 bar for 3 min, and the Improved Condition was preheated using liquefied petroleum gas and oxygen gas pressures of 1.2 bar and 4.5 bar for 6 min and 30 s. To investigate the mechanical and physical properties, micro-Vickers hardness tests, tensile tests and slow bending tests were also carried out. Welded metal in Normal Condition has many defects, including gas holes and shrinkage cavities. When comparing the Normal Condition to the Improved Condition, the Improved Condition demonstrates significantly more bending load and deflection. Specifically, the thermite welded rail sample of Improved Condition demonstrated a remarkable ability to endure bending loads of 108 tonnes and a deflection of 16 mm, and this sample remained unbroken until it exceeded 50% of the standardized deflection limit (10 mm). In addition, the average hardness values for the Improved Condition of the weld metal zone and the heat-affected zone were 331 HV and 289 HV, respectively. The Normal Condition produced an unsatisfactory fracture surface after slow bending test. This was caused by weld defects at the thermite weld due to inappropriate preheating.
Joint modeling of gene-environment correlations and interactions using polygenic risk scores in case-control studies
Authors: Wang Ziqiao , Shi Wen , Carroll Raymond J , Chatterjee Nilanjan
Published: May 2024

Polygenic risk scores (PRSs) are rapidly emerging as a way to measure disease risk by aggregating multiple genetic variants. Understanding the interplay of the PRS with environmental factors is critical for interpreting and applying PRSs in a wide variety of settings. We develop an efficient method for simultaneously modeling gene-environment correlations and interactions using the PRS in case-control studies. We use a logistic-normal regression modeling framework to specify the disease risk and PRS distribution in the underlying population and propose joint inference across the 2 models using the retrospective likelihood of the case-control data. Extensive simulation studies demonstrate the flexibility of the method in trading-off bias and efficiency for the estimation of various model parameters compared with standard logistic regression or a case-only analysis for gene-environment interactions, or a control-only analysis, for gene-environment correlations. Finally, using simulated case-control data sets within the UK Biobank study, we demonstrate the power of our method for its ability to recover results from the full prospective cohort for the detection of an interaction between long-term oral contraceptive use and the PRS on the risk of breast cancer. This method is computationally efficient and implemented in a user-friendly R package.
Plasma albumin, bilirubin, and uric acid and the subsequent risk of cancer: a case-cohort study in the Japan Public Health Center–based Prospective Study
Authors: Ihira Hikaru , Nakano Shiori , Yamaji Taiki , Katagiri Ryoko , Sawada Norie , Inoue Manami , Tsugane Shoichiro , Iwasaki Motoki
Published: May 2024

Several epidemiologic studies have investigated the circulating levels of albumin, bilirubin, and uric acid (UA) in relation to cancer risk; however, they have provided equivocal evidence. In this prospective case-cohort study, we measured the plasma levels of albumin, bilirubin, and UA and investigated their association with cancer incidence in 3584 case patients and 4270 randomly selected participants with a median follow-up of 15.8 years. The adjusted hazard ratios (HRs) and 95% CIs of total cancer for the highest quartile (Q4) versus lowest quartile (Q1) was 0.77 (95% CI, 0.67-0.90; P <.001 for trend) for albumin. This association was attenuated after excluding liver cancer cases with lower plasma albumin levels. Plasma bilirubin levels were positively related to liver cancer but inversely to total cancer after excluding liver cancer with, for Q4 versus Q1, an adjusted HR of 0.86 (95% CI, 0.74-0.99; P = .015 for trend). Plasma UA levels were not dose-responsively associated with total cancer risk. Higher plasma bilirubin levels were associated with a decreased risk of total cancer after excluding liver cancer, which is likely attributed to the antioxidant properties of bilirubin.
Suicide deaths involving opioid poisoning in the United States, by sex, 1999-2021
Authors: Hoopsick Rachel A , Yockey R Andrew , Campbell Benjamin M , Sauda Tonazzina H , Khan Tourna N
Published: May 2024

Suicide remains a leading cause of death in the United States, and recent data suggest suicide deaths involving opioids are increasing. Given unprecedented increases in drug-poisoning deaths, suicidality, and suicide deaths in recent years, an updated examination of the trends in suicide deaths involving opioids is warranted. In this descriptive epidemiologic analysis, we leverage final and provisional mortality data from the US Centers for Disease Control and Prevention’s WONDER database to examine trends in suicide deaths involving opioid poisoning from 1999 to 2021 by biological sex. Results reveal complex changes over time: the number and age-adjusted rate of suicide deaths involving opioid poisoning among male and female residents tended to track together, and both increased through 2010, but then diverged, with the number and rate of suicide deaths involving opioid poisoning among female residents outpacing that of male residents. However, the number and rate of suicide deaths involving opioid poisoning among male residents then began to stabilize, while that of female residents declined, closing the sex-based gap. Across all years of data, the proportion of suicide deaths that involved opioid poisoning was consistently higher among female decedents (5.8%-11.0%) compared with male decedents (1.4%-2.8%). Findings have implications for improved suicide prevention and harm reduction efforts.

This article is part of a Special Collection on Mental Health.
Antidepressant drugs and risk of developing glioma: a national registry-based case-control study and a meta-analysis
Authors: Malmberg Charlotte , Hellquist Barbro Numan , Sadanandan Sajna Anand , Sandström Maria , Wu Wendy Yi-Ying , Björkblom Benny , Melin Beatrice , Sjöberg Rickard L
Published: May 2024

Whether use of antidepressants is related to the risk of developing lower-grade (WHO grades 2-3) and higher-grade (WHO grade 4) glioma was investigated in this study. A registry-based case–control study was performed with 1283 glioma case patients and 6400 age-, sex-, and geographically matched control participants who were diagnosed in Sweden during 2009-2013. Conditional logistic regression was used to analyze whether selective serotonin reuptake inhibitors (SSRIs) or non-SSRIs were associated with the risk of developing lower- or higher-grade glioma in the study population. Our results show that use of antidepressant medication was not associated with the risk of developing glioma. We also performed a meta-analysis in which the data set from the present study was combined with results from 2 previous epidemiologic studies to answer the same questions. The meta-analysis showed a modest risk reduction of developing glioma in relation to antidepressant treatment (odds ratio = 0.90; 95% CI, 0.83-0.97) when all glioma subgroups and all forms of antidepressant medications were combined. In conclusion, it remains possible that antidepressants may have common monoaminergic mechanism(s) that reduce the risk of developing glioma.
Readon: a novel algorithm to identify read-through transcripts with long-read sequencing data
Authors: Chen Siang , Wang Hao , Zhang Dongdong , Chen Runsheng , Luo Jianjun
Published: May 2024

Motivation

There are many clustered transcriptionally active regions in the human genome, in which the transcription complex cannot immediately terminate transcription at the upstream gene termination site, but instead continues to transcribe intergenic regions and downstream genes, resulting in read-through transcripts. Several studies have demonstrated the regulatory roles of read-through transcripts in tumorigenesis and development. However, limited by the read length of next-generation sequencing, discovery of read-through transcripts has been slow. For long but also erroneous third-generation sequencing data, this study developed a novel minimizer sketch algorithm to accurately and quickly identify read-through transcripts.

Results

Readon initially splits the reference sequence into distinct active regions. It employs a sliding window approach within each region, calculates minimizers, and constructs the specialized structured arrays for query indexing. Following initial alignment anchor screening of candidate read-through transcripts, further confirmation steps are executed. Comparative assessments against existing software reveal Readon's superior performance on both simulated and validated real data. Additionally, two downstream tools are provided: one for predicting whether a read-through transcript is likely to undergo nonsense-mediated decay or encodes a protein, and another for visualizing splicing patterns.

Availability and implementation

Readon is freely available on GitHub (https://github.com/Bulabula45/Readon).
Characterization of senescence in human cardiac fibroblasts
Authors: Costa A , Ceresa D , Altieri P , Malatesta P , Ameri P
Published: May 2024

Funding Acknowledgements

None.

Background

Recent studies suggest that senescence of cardiac fibroblasts (cFib) plays a significant role in cardiac aging and disease. However, senescence of cultured cFib in response to classical pro-senescent stimuli has not been characterized yet.

Purpose

We thoroughly studied oncogene- and stress-induced senescence, as well as replicative senescence, in primary human cFib.

Methods

Human cFib were isolated from specimens of right atrial appendages, collected during cardiac surgery with extracorporeal circulation. These cells were subjected to oncogene-induced senescence by overexpression of HRasV12, were exposed to ionizing radiation (IR, 5 Gy) to recapitulate stress-induced senescence, and were passaged until replicative senescence was reached (P12-P14). Senescence was assessed by staining for senescence associated (SA) β-galactosidase and EdU and by RT-PCR for CDKN1A, CDKN2A, and LamB1.

Results

The main results are summarized in the graphical abstract. HRasV12 overexpression, IR, and prolonged culturing consistently caused senescence of cFib: in fact, all senescent models showed a significant increase in SA-β-gal activity and CDKN1A expression and a decrease in EdU uptake and LamB1 expression over control cells. Interestingly, cFib with HRasV12 overexpression exhibited a peculiar morphological phenotype with vacuoles and showed higher levels of the typical markers of senescence compared to control and to IR-treated or extensively passaged cFib. Moreover, only cFib with HRasV12 overexpression showed a strong downregulation of CDKN2A.

Conclusions

Cultured primary human cFib undergo senescence in response to established inducers of senescence. HRasV12 overexpression uniquely causes vacuolization of senescent cFib, which is currently being further characterized by transmission electron microscopy.
Exploiting spatial transcriptomics to investigate the molecular mechanisms involved in cardiac metastasis development
Authors: Lorizio D , Chiesa M , Pinamonti M , Bussani R , Ciucci G , Vuerich R , Zacchigna S
Published: May 2024

Funding Acknowledgements

Type of funding sources: Foundation. Main funding source(s): IEO-Monzino Foundation

Background

Cardiac metastasis is considered a relatively rare malignancy, although an increasing prevalence is expected in the future due to the increasing life expectancy. Several cases of cardiac metastases remain untreatable, given the anatomical location that often hampers both surgical eradication and bioptic collection for biological characterization. Here, we use spatial transcriptomics to provide an unbiased molecular characterization of cardiac metastases, and identify novel targets for their treatment.

Material and methods

We performed spatial transcriptomics on samples of primary tumor, cardiac, and extra-cardiac metastasis from three different patients. First, we checked RNA integrity by means of DV200 analysis. Spatial transcriptomic analysis was performed by using the GeoMX technology (Nanostring), which provides whole-transcriptome data along with spatial distribution information of the preselected samples from the candidate regions of interest (ROIs). ROIs were selected on the formalin-fixed paraffin-embedded sections by means of geometric shapes in combination with cell markers specific for tumor cells, cardiomyocytes and endothelial cells, respectively. Differential gene expression analysis and Gene Set Enrichment Analysis (GSEA) were performed on the entire transcriptome. Cell-cell interactions between cardiac cells and cancer cells were then evaluated by means of InterCellaR.

Results

Differential expression analysis between cardiac metastases and extra-cardiac tumors (primary tumors + extra-cardiac metastases) identified 92 differentially expressed genes (DEGs). Of these, 53 genes were significantly overexpressed in cardiac metastases, while 39 genes were overexpressed in the extra-cardiac samples, regardless of the site of origin of the tumor. Unsupervised hierarchical clustering of protein-coding genes clearly separated the samples according to their anatomical location, in particular cardiac metastasis from extra-cardiac tumors. GSEA identified several pathways enriched in cardiac metastases compared to the extra-cardiac tumors, including mechanotransduction (e.g. keratinization and cell-matrix interaction) and triglyceride metabolism, suggesting a potential role of these signaling pathways in the development of cardiac metastasis. Interactome analysis revealed a rich plethora of receptor-ligand interactions in cardiac metastasis, especially between tumor cells and cardiomyocytes.

Conclusions

These results start shedding light on the mechanisms that allow cancer cell growth in the heart and pave the way to the development of novel therapies to treat this malignancy.
The endocannabinoid ARA-S acts as an endogenous rescuing factor of Kv7.1/KCNE1 carrying Long QT syndrome mutations and paves the way for drug development strategies
Authors: Hiniesto Inigo I , Sridhar A , Lundholm S A , Liin S I
Published: May 2024

Funding Acknowledgements 

Type of funding sources: Public grant(s) – EU funding. Main funding source(s): European Union’s Horizon 2020 research and innovation program

Long QT Syndrome (LQTS) is a cardiac arrhythmia characterized by delayed ventricular repolarization. LQTS occurs when there are loss-of-function mutations in genes encoding for voltage-gated channels and their beta subunits that contribute to the cardiac action potential. Among them, the most mutated channel is Kv7.1/KCNE1, which encodes for the slowly activating component of the delayed rectifier K+ current (IKs). Families with inherited mutations in the Kv7.1/KCNE1 exhibit inconsistent correlations between genotypes and phenotypes making the clinical management of LQTS challenging. A possible strategy to help guide clinical approaches involves exploring endogenous compounds capable of modulating the Kv7.1/KCNE1 channel function, acting as potential disease modifiers. Using the two-electrode voltage clamp technique, we previously found that the endogenous endocannabinoid-like compound ARA-S, which has an arachidonic acid tail and a serine head group, facilitates activation of the wild-type Kv7.1/KCNE1 channel expressed in Xenopus oocytes. This is seen as a left shift in the voltage dependence of channel opening and an increase in conductance. Moreover, we showed that ARA-S restores a physiological QT interval in isolated guinea pig hearts suffering from drug-induced LQTS. Hence, ARA-S is an interesting endogenous Kv7.1/KCNE1 channel modulator that may influence disease severity. However, it is uncertain whether ARA-S also activates the Kv7.1/KCNE1 when it carries LQTS-associated mutations. Therefore, the current study involves studying the effect of ARA-S on mutated Kv7.1/KCNE1 channels using two-electrode voltage clamp electrophysiology. We find that ARA-S facilitates the activation of all tested LQTS mutants. However, the magnitude of the effect varies among them, suggesting that the compound´s effect is dependent on the genetic background. To understand why ARA-S has different effects, we use computational simulation approaches to study how mutations in the putative ARA-S binding sites change ARA-S efficacy. This understanding aids in developing new substances that can activate different mutations, by, for example, modifying the ARA-S tail to enhance its efficacy. We also try to understand the concentration of ARA-S required to restore a wild-type-like function of each mutant, considering the mutant's intrinsic behavior and the apparent affinity of ARA-S for the specific mutant. Altogether, this work finds ARA-S to be an efficient activator of LQTS-associated mutant Kv7.1/KCNE1 channels and is a first step towards understanding how the genetic background determines the effect of this endogenous modulator.
Real-time (re)shaping of cardiac action potentials by an opto-electronic feedback control system in the multicellular setting
Authors: Ordog B , De Coster T , Bart C I , Zhang J , Boink G J J , Bax W , Deng S , Den Ouden B L , De Vries A A F , Pijnappels D A
Published: May 2024

Funding Acknowledgements

Type of funding sources: Public grant(s) – EU funding. Main funding source(s): European Research Council Starting Grant and Consolidator Grant to Daniël A Pijnappels

Background

The membrane potential (Vm) in cardiomyocytes fulfills essential regulatory roles in various biological functions, including not only action potential (AP) propagation and mechanical contraction, but also homeostatic regulation. In-depth studies into these roles have been severely hampered by the lack of research methods allowing full control over Vm, especially in multicellular cardiac preparations.

Purpose

We aimed to unlock new research possibilities by the development of an experimental system (APqr) capable of full Vm control, including instantaneous modulation of AP morphology on a multicellular levelin the multicellular setting.

Methods

Monolayers of immortalized human atrial myocytes (hiAMs, n=7) were genetically modified to express the blue light-activatable cation channel Cheriff and the red light-sensitive inward chloride pump Jaws for depolarizing and re- or hyperpolarizing effects, respectively. Real-time Vm readout was obtained by patch clamp electrophysiology. Deviations of Vm from reference values were calculated by a custom closed-coop controller. A 470-nm and a 617-nm LED were modulated by the controller in a Vm deviation-dependent manner, selectively activating Cheriff or Jaws. Electrical disturbances were introduced by application of 4-aminopyridien (4AP, 200 µM) or a preprogrammed blue-light pulse creating abnormal cation influx during the repolarization phase of the hiAM AP.

Results

Average AP durations at 90% repolarization (APD90) increased by 82.4 ms following 4AP application and by 362.5 ms in the presence of light-induced abnormal cation influx compared to control conditions. APqr reduced these APD90 differences to 1.9 ms and 4.6 ms on average, with Vm deviations less than 2.5 mV in 74.7% and 94.4% of the time, respectively. APqr could also be applied for the enforcement of arbitrary AP shapes. In these experiments, reference Vm values consisted of APs recorded from hiAM monolayers exposed to drugs with APD-prolonging (4AP) or shortening (carbachol) effects. APqr was able to enforce these reference APs on hiAM monolayers with high accuracy, with Vm deviations less than 2.5 mV in 95.8% and 94.4% of the time, respectively.

Conclusions

APqr preserves AP morphologies in the presence of electrical perturbations of different origin without any prior knowledge of the disturbance and enforces arbitrary AP morphologies with high accuracy in an immediate and self-regulatory manner. Collectively, these results set the stage for the refinement and application of opto-electronic control systems to enable in-depth investigation into the regulatory roles of the membrane potential in health and disease.
Modeling doxorubicin-induced cardiotoxicity with human myocardial tissue slices: insights on genetic susceptibility and cardioprotection
Authors: Van Der Geest J S A , Kelters I R , Van Der Kaaij N P , Dendorfer A , Doevendans P A F M , De Boer T P , Van Laake L W , Sluijter J P G , Sampaio Pinto V
Published: May 2024

Funding Acknowledgements

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): PSIDER and HARVEY (ZonMW)

Introduction

Cardiotoxicity poses a significant challenge in drug development, often leading to the withdrawal of approved drugs. To address this issue, there is a critical need for improved models to predict cardiac adverse effects. In line with the imperative for animal-free preclinical testing, this study aimed to develop a novel model using human myocardial tissue slices to get a deeper understanding and potential prevention of doxorubicin-Induced cardiotoxicity.

Methods

Human myocardial tissue slices of 300 μm thick were derived from end-stage heart failure patients explanted hearts, including those harbouring likely or pathogenic cardiovascular mutations (class 4 and 5) (N=3) and non-genetic cardiovascular etiology (N=3). Myocardial tissue slices were cultured under physiological mechanical and electrical conditions. Slices were exposed to doxorubicin (1 µM) over a 10-day culture period, and their responses were compared to DMSO-vehicle controls, by recording contractile force and Ca2+ transients. Additionally, a subset of slices was treated with a cardioprotective agent, dexrazoxane (100 µM), 1 day prior to and during doxorubicin exposure to assess potential cardioprotection.

Results

Doxorubicin-exposed slices with a pathogenic mutation exhibited reduced mechanical force, increased threshold potential, and impaired ability to follow pacing at higher frequencies. Additionally, aberrations in calcium handling, manifested as arrhythmia-like disconnection of muscle fibers, were notably observed in these slices. Instead, slices from patients without pathogenic mutations showed minimal cardiotoxicity when exposed to doxorubicin. Encouragingly, pretreatment with dexrazoxane demonstrated a protective effect against doxorubicin-induced cardiotoxicity in the genetically susceptible group. Our functional findings corroborated our structural data, highlighting the breakdown of sarcomere structure and cell-cell connections and increased DNA damage, particularly in patients harbouring a genetic cardiovascular mutation.

Conclusion

In summary, human myocardial tissue slices faithfully recapitulate doxorubicin-induced cardiotoxicity. Importantly, our data suggests the utility of employing genetic testing for known cardiovascular mutations as a valuable step before initiating doxorubicin treatment. Additionally, we demonstrate for the first time the in vitro cardioprotective potential of dexrazoxane in a human tissue model. These findings pave the way for using ex vivo myocardial tissue slices to evaluate cardiotoxicity and their value as a powerful tool for evaluating the efficacy and delivery approaches.
Cardiovascular toxicity profile of doxorubicin: data mining of the FDA adverse event reporting system (FAERS)
Authors: Kattan L , Cerbito E , Dhulkifle H , Korashy H , Maayah Z
Published: May 2024

Funding Acknowledgements

None.

Background

Doxorubicin (DOX), an effective anticancer agent, is associated with dose-dependent cardiovascular toxicity. Understanding its mechanisms and risk factors will facilitate novel interventions to minimize DOX-induced cardiovascular toxicity.

Purpose

We aimed to perform a comprehensive analysis of the Food and Drug Administration Adverse Event Reporting System (FAERS) database to identify, describe, and quantify the latest cardiovascular DOX-associated adverse-events (AEs).

Methods

Data from 2004 Q1 – 2022 Q3, where DOX was the primary-suspect, were extracted from FAERS via OpenVigil 2.1. For data collection, preferred-terms (PTs) from Medical Dictionary for Regulatory Activities (MedDRA) were used, while ROR with 95% CIs for disproportionality analysis was performed.

Results

1987 reports were collected for significant PTs. The most prevalent DOX-associated AE was heart failure (77.6%, n=1542), followed by cardiac fibrosis and inflammation (7.75%, n=154) and vascular disorders (7.45%, n=148). Others include arrhythmias, systemic inflammation, and endothelial dysfunction. Most patients were females (51.1%, n=1016), 18-64 years-old (41%, n = 815), primarily from Europe (41.87%, n=832) and the United States (34.12%, n = 678). Six outcomes were associated with DOX-induced cardiotoxicities, mainly "Death", "Initial/prolonged hospitalization", and "Others".

Conclusion

This study provides valuable insights on DOX-induced cardiovascular toxicity using real-world data from FAERS.Total Case Count of all Adverse EventPatients' Demographics and outcomes
AAV-mediated gene delivery to the cardiac conduction system
Authors: Jonker T , Boender A R , Klerk M , Barnett P , De Vries A A F , Christoffels V M , Boink G J J
Published: May 2024

Funding Acknowledgements

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): NWO open technology program Biopace #18485

Background

Gene therapy for cardiac arrhythmias has so far mostly focused on modifying the arrhythmogenic substrate in working myocardium or on creating biological pacemakers with viral vectors that did not selectively target the cardiac conduction system (CCS). As a result, CCS targeting by viral vectors is a relatively unexplored area.

Aim

In the current project, we explored the basic requirements for CCS targeting using adeno-associated virus (AAV) vectors.

Methods

Regular AAV9 and AAV-Myo4A capsid-pseudotyped AAV2 vectors (hereafter called AAV2/9 and AAV2/Myo4A vectors, respectively) expressing enhanced green fluorescent protein (eGFP) from a CMV or TNNT2 promoter, respectively, and an AAV2/9 vector expressing eGFP from a human GJA5 regulatory element were systemically injected in mice. After 4 weeks, hearts were harvested, fixed with paraformaldehyde and stained for TNNT2, eGFP, HCN4 and DNA to determine CCS transduction.

Results

Transduction of the sinoatrial and atrioventricular (AV) node using AAV2/9 vectors was very poor, while transduction was visible in the atrioventricular (i.e. His) bundle and working myocardium. In contrast, the transduction rate of the AV node and bundle using AAV2/Myo4A vectors were similar to that of the working myocardium.

The human GJA5 regulatory element was able to restrict eGFP expression by AAV2/9 vectors to the AV bundle in 2/3 animals but the transduction efficiency was low. A third animal showed no transduction of any tissue, either indicative of the low transduction efficiency or of a failed injection.

Conclusion

This study not only provides proof-of-principle for CCS-targeted gene therapy but also shows that CCS transduction can be improved through AAV capsid engineering and be restricted to (parts of) the CCS by using specific transcriptional control elements.
Transcriptomic insights into cardiac healing - unveiling promising targets for effective gene therapies in diseased hearts
Authors: Caliandro R , Hanemaaijer J , Van Den Berg S , Gunst Q , Oostra R J , Van Den Hoff M , Gladka M M
Published: May 2024

Funding Acknowledgements 

Type of funding sources: Foundation. Main funding source(s): Dutch Heart Foundation

Identifying therapeutical target genes is the first step towards developing new gene therapies to promote cardiac repair after injury. In the last decades, despite the many steps forward, not a single therapy has shown to be effective and suitable for human treatment of the injured heart. Considering that cardiac diseases represent one of the most prevalent causes of morbidity, disability, and mortality in the world, it is, therefore, essential to develop new and improved therapies to induce cardiac regeneration.

In this study, we aimed to identify novel therapeutical targets that could be used for future purposes of cardiac gene therapy. Unlike previous studies that investigated fetal hearts that are physiologically and functionally different from mature hearts, we focused on investigating adult human hearts that we collected from human donors with post-mortem time below 5 hours. Samples of left ventricles were molecularly and morphologically characterized for the presence of stress, cardiac damage, and cardiac fibrosis. Results of the molecular analyses were cross-referenced with the donors' medical history to classify each specimen as "control" or "diseased." Next, we performed single nuclei RNA sequencing of samples of the left ventricle from 2 "control" and 3 "diseased" hearts. Due to the adult tissue's complexity and the specimen's high-fiber content, we optimized the nuclei isolation protocol. Eventually, we successfully sequenced an average of 8.000 nuclei per sample with a gene coverage of approximately 800 genes per nucleus, identifying, via unsupervised cluster analysis, 17 different cell populations.

In cardiomyocytes, we identified over 500 genes differentially expressed in diseased hearts when compared to healthy controls. Among these genes, we discovered several novel target genes whose expression is increased only in diseased cardiomyocytes. Gene ontology analysis of the known interactors of the newly identified target genes shows enrichment in terms such as sarcomere organization, muscle contraction, mitochondrial energetics, calcium handling, and fatty acid metabolism.

Currently, we are investigating the molecular effects of the gain and loss of function of these genes in healthy iPS-derived cardiomyocytes and human-derived myocardial tissue slices to better understand their role in the pathophysiology of the adult human heart. From a therapeutic perspective, our ongoing research holds promises for developing new and improved intervention therapies to promote cardiac regeneration.
HCN2-based biological pacing in a modified rat AV-block model
Authors: Jonker T , Park K , Schroedel S , Neef K , Klerk M , Kirzner O F , Tan H L , Christoffels V M , Boink G J J
Published: May 2024

Funding Acknowledgements

Type of funding sources: Public grant(s) – EU funding. Main funding source(s): Eurostars - Heartpace 114245

Background

Electronic pacemakers have several important shortcomings which are difficult to overcome due to their hardware-based design. Gene therapy-based biological pacemakers may provide a different platform which can overcome these limitations. Such biological pacemaker strategies center around overexpression of the pacemaker ion channel HCN2, either alone or with other transgenes to potentiate pacemaker performance, such as the skeletal muscle sodium channel SkM1. In order to develop clinically applicable pacemaker gene therapies further engineering is needed, which would benefit from a reliable small animal model. At present it is unknown if the rat AV block model can be used to evaluate ion channel-based biological pacing.

Aim

In the present study, we aim to validate a modified rat AV-block model for the evaluation of HCN2 and SKM1-based biological pacing.

Methods

AV-block was generated in female rats by epicardially applied needle ablation of the AV-nodal region, as previously described (N.K. ). Complete AV-block was confirmed 7 days post-ablation by loss of P-wave and QRS-complex association on the body surface ECG. Seven days post-ablation, 3 groups of animals were injected into the left ventricular apex with Ad5-TNNT2-HCN2, Ad5-TNNT2-SkM1 or saline (control). 14 days post-ablation, ECG analysis was performed during anesthesia under baseline and after i.v. administration of isoprenaline. Upon completion of functional testing, tissue samples were harvested followed by gene expression analysis using qPCR.

Results

Injection of Ad5-TNNT2-HCN2, Ad5-TNNT2-SkM1 or saline did not result in ectopic activation patterns in baseline conditions 7 days after injection. Administration of isoprenaline results in reversal of the QRS complexes 90% of total measurement time in Ad5-TNNT2-HCN2 injected animals. Saline and Ad5-TNNT2-SkM1 showed no change in activation pattern upon isoprenaline administration. Gene expression analysis revealed expression of HCN2 in the injection site following Ad5-TNNT2-HCN2 injection, with minimal spread to non-target regions. However, this analysis also showed Ad5-TNNT2-SkM1 did not express well in this context.

Conclusion

Our modified rat AV-block model, when administered with isoprenaline, recapitulated expected biological pacemaker activity of HCN2 gene delivery. However, lack of effective expression of SkM1 renders direct comparison of these relevant transgenes difficult. Nevertheless, we expect this rat AV-block model can provide a valuable tool for the evaluation of novel ion channel-based biological pacemaker strategies.
Early biomarkers of disease in hypertrophic cardiomyopathy: does telomere length matter?
Authors: Judez Serrano A , Louchachha Medhi S , Munteanu S E M , Nicolas Rocamora M E , Sabater Molina M , Gimeno Blanes J R
Published: May 2024

Funding Acknowledgements

Type of funding sources: Public Institution(s). Main funding source(s): Biomedical Research Institute of Murcia (IMIB-Arrixaca)

Introduction

Hypertrophic cardiomyopathy (HCM) is the leading cause of sudden death in people under 35 years of age. (1) The variability of the phenotype together with its incomplete penetrance has been an obstacle to a complete understanding of the clinical spectrum and consequences of the disease. In recent years, there has been increasing interest in telomeric shortening and several authors have linked it to age-related diseases and HCM. (2, 3) Our aim was to study whether telomere length could affect the age at presentation and the severity of the HCM phenotype in genetic carriers.

Methods

Measurement of telomere length in circulating leukocytes (LTLc) was performed in 96 patients carrying a mutation in MYBPC3 causing HCM [58 affected (24 severely affected), 38 unaffected]. The variants included were c.2308+1G>A, p.Pro108Alafs*9, p.Arg891Alafs*160 and p.Glu258Lys, all considered founder mutations in our region.

Peripheral venous blood samples were obtained from patients at the Inherited Heart Diseases Unit and stored at -80 ºC after informed consent had been signed by all participants.

For telomere length analysis, a telomeric restriction fragment (TRF) assay was performed using the TeloTAGGGTelomereLenghtAssay kit (Roche, 12209136001) according to the manufacturer's protocol with slight modifications.

Results

Our results show a lower LTLc in those affected patients with a more severe HCM phenotype compared to the affected and unaffected group (p=0.023) independently of age and sex, which are two variables that affect LTLc. Similarly, a shorter telomere length was associated with greater severity of the phenotype, with patients presenting a larger left atrial size (p=0.032), septal size (p=0.023) and greater obstruction (p=0.044). The presence of atrial fibrillation (p=0.039) and sudden death (p=0.024) was also higher in patients with shorter telomeres.

Conclusions

Telomere length is a variable that influences the expression of the phenotype in HCM patients carrying a MYBPC3 variant. This is an observation that supports the thesis of numerous publications linking HCM and other age-related diseases, sometimes called telomeropathies,(2) to telomere shortening. It has also been linked to non-hereditary conditions(3).

This fact sheds some light on the complex phenotypic variability present in HCM, raising LTL as a promising prognostic biomarker of the disease, as has already been suggested by other authors.(4, 5, 6).Workflow in the telomere legth analysisClinical associations of LTLc
Swimming training induced sex-specific differences in cardiac hypertrophy and blood levels of the renin-angiotensin system
Authors: Otmani Idrissi M O I , Frobert F A , De Chambrier A C , Stumpe M S , Fluck M F , Nikodemski M N , Giraud M N G
Published: May 2024

Funding Acknowledgements

Type of funding sources: Foundation. Main funding source(s): FNS

Background

Exercise promotes physiological cardiac hypertrophy that is adaptive and reversible and influenced by age, genetics, sex, exercise intensity and type. The renin-angiotensin system (RAS) plays a crucial role in cardiovascular remodelling at both systemic and tissue levels. However, a significant gap exists in our understanding of sex differences in response to physical activity. Our objective is to characterise swimming-induced cardiac remodelling in female and male rats and RAS at both the cardiac tissue and systemic levels.

Methods

Male and female Lewis rats swam in a container filled with tap water at 32° C for 1 hour per day for 4 weeks or up to 8 weeks, respectively. Sedentary animals served as control. Echocardiography and immunostaining of RAS components, including angiotensin-converting enzyme 2 (ACE2) on cardiac sections' staining were performed. Cardiomyocyte (CM) size was measured. Cardiac RAS receptors were quantified with RT-PCR. Blood levels of Angiotensinogen, Angiotensins (Ang) were measured with Mass Spectrometry. Renin activity was calculated by the ratio Ang1/Angiotensinogen.

Results

After 4 weeks, an increase in ventricle diameter was shown in exercised male rats compared to the control. The cardiac hypertrophy was confirmed by increased heart/body weight (H/BW= 3x10-3±1.3x10-4vs2.5x10-3±3.10-5mg/g p<0.0001) and CM size (288±23 vs 198±1μm2 p=0.0028) associated with increased intra-CM immunostained ACE2 (AC2 area fraction: 38.5%±2.8%vs20%±5 p=0.0001). In females, ventricle diameter was unchanged, but wall thickness increased. Exercise was pursued for up to 8 weeks. At 6 and 8 weeks, echocardiography measurements remained similar as at 4 weeks. Hypertrophy was then confirmed by increased H/BW (3.2x10-3±1.7x10-4vs control:2.7x103 ±1.8x104mg/g p<0.0001) and CM size (335±47vs233±25μm2 p= 0.0019). An increased intra-CM-ACE2 was characterised by immunostaining (55.8%±6vs32.5%±6.6% p<0.0001) and confirmed by the increased ACE2 gene expression in cardiac tissue.

At 4 weeks, the blood levels of RAS components were similar in trained and control males as well as renin activity. On contrary, exercised female rats showed increased blood levels of Angiotensinogen (4.2x10-4±7x10-5 vs 3x10-4±7x10-5au p=0.0288), Ang1 (2±1.9vs0.0±0.9au p=0.0151), Ang2 (1.2±1.5vs0.0±0.9au p=0.0204), and Ang1-7 (3.4±2.9vs 0.0±0.9au p=0.0058). The blood levels of these RAS compounds returned to the control levels after 6 and 8 weeks.

In addition, a sex-distinct blood proteome was identified.

Conclusion

Our results showed a sex-distinct RAS regulation post-exercise and suggested a sex difference in the timing of adaptive responses at the system and cardiac tissue levels.
Soluble guanylate cyclase stimulator as an emerging therapeutic option in the management of arrhythmias in heart failure rat model
Authors: Szeiffova Bacova B , Martiskova A , Sykora M , Gawrys O , Tribulova N , Cervenka L
Published: May 2024

Funding Acknowledgements 

Type of funding sources: Foundation. Main funding source(s): This research was supported by VEGA grant no. 2/0002/20 and 2/0006/23, the Slovak Research and Development Agency under contract no. 21-0410, European Regional Development Fund: ITMS2014+: 313011AVG3. This research was also partially supported by the Ministry of Health of the Czech Republic within the project for the development of the research organization [grant number 00023001 (IKEM)—institutional support] and also supported by the project National Institute for Research of Metabolic and Cardiovascular Diseases (Program EXCELES, Project No. LX22NPO5104)—Funded by the European Union—Next Generation EU

Heart failure (HF) is a complex clinical syndrome characterized by a decrease in the efficiency of the heart pump, leading to a decline in hemodynamic status and cardiac remodelling, that can contribute to the formation of a substrate for arrhythmias. Antiarrhythmic drug therapy is still suboptimal and current therapies are not efficacious enough. The purpose of the current study was therefore to examine the efficiency of nitric oxide (NO)/soluble guanylyl cyclase (sGC)/cyclic guanosine monophosphate (cGMP) stimulator.

We used as an experimental model male hypertensive Ren-2 transgenic rats (TGR, n = 16), normotensive Hannover Sprague–Dawley rats (HSD, n = 17), and a rat model of HF surgically induced by aortocaval fistula (ACF) in rats of both strains (n = 27). After ACF induction were rats treated by sGC stimulator BAY41-8543 (3 mg/kg/day for 30 weeks) alone or combined with angiotensin-converting enzyme inhibitor (ACEi) Trandolapril (0.25 mg/kg/day for 30 weeks). Age of the animals at the end of the experiment was 40 weeks. Left ventricle tissue and plasma samples were used for further analyses.

Treatment by sGC stimulator significantly decreased rats' mortality, and systolic blood pressure, and significantly increased levels of antioxidant enzyme superoxide dismutase 1, matrix metalloproteinase 2, total and phosphorylated connexin 43 as well as protein kinase C epsilon, implicated in remodelling of extracellular matrix and intercellular communication, in the left ventricle of TGR rats. Application of sGC stimulator slightly normalized elevated collagen deposition and hydroxyproline content in the left ventricular tissue of TGR rats. Unfortunately, we did not observe any effect of the sGC stimulator in the TGR ACF group.

These results support the hypothesis that sGC stimulators might represent a class of drugs suitable for combating heart failure with partially anti-arrhythmic potential, but further studies are needed.