bluuebunny/crossref_metadata_2025_split huggingface dataset:
Secular trends in adherence to healthy behaviors among US adults with diabetes, 1999-2018
Authors: Feng Chengwu , Feng Li , Shan Zhilei , Han Han , Cao Yaying , Sheng Aili , Guasch-Ferré Marta , Geng Xin , Wang Liang , Zong Geng 
Published: May 2024

Abstract
Adherence to a healthy lifestyle is essential for diabetes management in light of the plateaued metabolic control, diversified causes of death, and continued excess mortality among people with diabetes (PWD). In this study, the secular trend of adherence to healthy behaviors was assessed among PWD in the National Health and Nutrition Examination Survey, a nationally representative survey of Americans, begun in 1999, that uses a stratified, multistage probability design in 2-year cycles. Adherence to a healthy lifestyle was estimated via a scale using never smoking, moderate drinking, adequate physical activity, and healthy diet data; the scores range from 0 to 4. Among 7410 participants, adherence to healthy behaviors across time slightly increased from a score of 1.4 (95% CI, 1.3-1.5) in 1999-2002 to 1.6 (1.5-1.8) in 2015-2018 (P = .002 for trend). The overall lifestyle score of non-Hispanic Black people increased to the same score as non-Hispanic White people (1.7 vs 1.6 in 2015-2018). Large socioeconomic disparities remained in that participants with higher income and education level and who were covered by health insurance were more likely to adhere to healthy behaviors. The metabolic control plateaued and causes of death have diversified among PWD; thus, our findings suggested a potential for lifestyle modification in facilitating the long-term health of these patients.
A hypothetical intervention to reduce inequities in anxiety for Multiracial people: simulating an intervention on childhood adversity
Authors: Lam-Hine Tracy , Bradshaw Patrick , Allen Amani , Omi Michael , Riddell Corinne 
Published: May 2024

Abstract
Multiracial people report higher mean Adverse Childhood Experience (ACE) scores and prevalence of anxiety than other racial groups. Studies using statistical interactions to test if associations between ACEs and anxiety are greater for this group than others have shown mixed results. Using data from waves 1 (1995-1997) through 4 (2008-2009) of the National Longitudinal Study of Adolescent to Adult Health (Add Health), we simulated a stochastic intervention over 1000 resampled datasets to estimate the race-specific cases averted per 1000 of anxiety if all racial groups had the same exposure distribution of ACEs as Whites. Simulated cases averted were greatest for the Multiracial group, (median = −4.17 cases per 1000; 95% CI; −7.42 to −1.86). The model also predicted smaller risk reductions for Black participants (−0.76; 95% CI, −1.53 to −0.19). CIs around estimates for other racial groups included the null. An intervention to reduce racial disparities in exposure to ACEs could help reduce the inequitable burden of anxiety on the Multiracial population. Stochastic methods support consequentialist approaches to racial health equity, and can encourage greater dialogue between public health researchers, policymakers, and practitioners.
This article is part of a Special Collection on Mental Health.
Temperature dependence of pollen germination and tube growth in conifers relates to their distribution along an elevational gradient in Washington State, USA
Authors: Hsu Hsin-Wu , Kim Soo-Hyung 
Published: May 2024

Abstract

Background and Aims
Pollen germination and tube growth are essential processes for successful fertilization. They are among the most temperature-vulnerable stages and subsequently affect seed production and determine population persistence and species distribution under climate change. Our study aims to investigate intra- and interspecific variations in the temperature dependence of pollen germination and tube length growth and to explore how these variations differ for pollen from elevational gradients.

Methods
We focused on three conifer species, Pinus contorta, Picea engelmannii and Pinus ponderosa, with pollen collected from 350 to 2200 m elevation in Washington State, USA. We conducted pollen viability tests at temperatures from 5 to 40 °C in 5 °C intervals. After testing for 4 d, we took images of these samples under a microscope to monitor pollen germination percentage (GP) and tube length (TL). We applied the gamma function to describe the temperature dependence of GP and TL and estimated key parameters, including the optimal temperature for GP (Topt_GP) and TL (Topt_TL).

Key Results
Results showed that pollen from three species and different elevations within a species have different GP, TL, Topt_GP, and Topt_TL. The population with a higher Topt_GP would also have a higher Topt_TL, while Topt_TL was generally higher than Topt_GP, i.e. a positive but not one-to-one relationship. However, only Pinus contorta showed that populations from higher elevations have lower Topt_GP and Topt_TL and vice versa. The variability in GP increased at extreme temperatures, whereas the variability in TL was greatest near Topt_TL.

Conclusions
Our study demonstrates the temperature dependences of three conifers across a wide range of temperatures. Pollen germination and tube growth are highly sensitive to temperature conditions and vary among species and elevations, affecting their reproduction success during warming. Our findings can provide valuable insights to advance our understanding of how conifer pollen responds to rising temperatures.
IntelliGenes: Interactive and user-friendly multimodal AI/ML application for biomarker discovery and predictive medicine
Authors: Narayanan Rishabh , DeGroat William , Mendhe Dinesh , Abdelhalim Habiba , Ahmed Zeeshan 
Published: May 2024

AbstractArtificial intelligence (AI) and machine learning (ML) have advanced in several areas and fields of life; however, its progress in the field of multi-omics is not matching the levels others have attained. Challenges include but are not limited to the handling and analysis of high volumes of complex multi-omics data, and the expertise needed to implement and execute AI/ML approaches. In this article, we present IntelliGenes, an interactive, customizable, cross-platform, and user-friendly AI/ML application for multi-omics data exploration to discover novel biomarkers and predict rare, common, and complex diseases. The implemented methodology is based on a nexus of conventional statistical techniques and cutting-edge ML algorithms, which outperforms single algorithms and result in enhanced accuracy. The interactive and cross-platform graphical user interface of IntelliGenes is divided into three main sections: (i) Data Manager, (ii) AI/ML Analysis, and (iii) Visualization. Data Manager supports the user in loading and customizing the input data and list of existing biomarkers. AI/ML Analysis allows the user to apply default combinations of statistical and ML algorithms, as well as customize and create new AI/ML pipelines. Visualization provides options to interpret a diverse set of produced results, including performance metrics, disease predictions, and various charts. The performance of IntelliGenes has been successfully tested at variable in-house and peer-reviewed studies, and was able to correctly classify individuals as patients and predict disease with high accuracy. It stands apart primarily in its simplicity in use for nontechnical users and its emphasis on generating interpretable visualizations. We have designed and implemented IntelliGenes in a way that a user with or without computational background can apply AI/ML approaches to discover novel biomarkers and predict diseases.
Dissecting signal transduction pathways that regulate cardiomyocyte proliferation
Authors: Artioli R , Cannilla R , Dal Ferro M , Zentilin L , Giacca M , Collesi C 
Published: May 2024

Abstract

Funding Acknowledgements
None.

Introduction
During cardiac development, the Notch, Hippo, and Wnt/β-catenin signalling pathways play a crucial role in the tuning of cardiomyocyte proliferation. The individual engagement of each of these pathways during the early postnatal life, however, is still unclear.

Purpose
We wanted to develop a method to visualize over time, both in vitro and in vivo, the topography of activation of the Notch, Wnt/β-catenin, and Hippo pathways and their crosstalk upon cardiomyocyte treatment with a series of pro-proliferative stimuli.

Methods
We obtained Adeno-Associated Virus (AAV) constructs expressing distinct fluorescent proteins (ECFP, GFP, mCherry, DsRed) under the control of promoters that are specifically activated downstream of the signalling cascades of interest (a synthetic promoter consisting of 12 CSL binding sites for Notch1, a synthetic promoter consisting of 8 TEAD binding sites for YAP, and a synthetic promoter containing 7 TCF/LEF1 binding sites for β-catenin).

Results
First, by high content microscopy we followed the time of the activation of the AAV vectors coding for fluorescent reporters, either alone or in combination. We observed that a crosstalk exists between Yap and Notch signalling in proliferating cardiomyocytes, as detected by the activation of their respective fluorescent reporters. We confirmed our data by gene expression analysis and co-immunoprecipitation using specific antibodies in fractionated cardiomyocyte extracts. Next, we observe that miR-1825, miR-199a-3p, and miR-590-3p – three pro-regenerative miRNAs in cardiomyocytes– specifically activate both Yap and Notch signalling. Finally we characterized the reporter systems in vivo, to dissect the intracellular signalling pathways that are active in the first week of postnatal life. Beta-catenin signalling was quite undetectable in any of the conditions tested. In contrast, both Notch and Yap showed significant activation, consistent with their role in regulating early postnatal cardiomyocyte proliferation.

Conclusions
The developed AAV-fluorescent reporters allow to efficiently and specifically visualize the pro-proliferative activation of Notch and Yap signalling both in vitro and in vivo. The results obtained are supportive of a crosstalk existing between Notch and Yap pathways in proliferating cardiomyocytes.
Propafenone-induced connexin-43 protein accumulation and intracellular dynamics
Authors: Li E , Boujeddaine N , Mol B , Van Der Heyden M A G 
Published: May 2024

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): China Scholarship Council

Background
Connexin-43 (Cx43) plays a pivotal role in intercellular communication through gap junctions. It is instrumental in propagating electrical signals and allowing ions to flow between neighboring cardiomyocytes. This synchronized electrical activity allows the heart to work as a functional syncytium. Any disruption or malfunction of Cx43 can lead to arrhythmias and other cardiac issues. Understanding the functions and regulation of Cx43 is vital in both basic research and clinical contexts. Propafenone, a class Ic antiarrhythmic drug, has shown promise in rhythm control; however, its precise impact on cardiac cellular physiology, particularly regarding Cx43, remains incompletely understood.

Purpose
To investigate propafenone’s effect on Cx43 expression, physiology, and underlying mechanisms in cell systems.

Methods
Used cell lines include human embryonic kidney HEK293 cells transfected with Cx43; differentiated murine embryonic carcinoma EPI7 cells with an epithelioid morphology and visceral endoderm-like END2 cells, endogenously expressing high amounts of functional Cx43. Cx43 protein expression levels were determined by Western blot analysis, whereas immunofluorescence (IF) imaged by a Total Internal Reflection Fluorescence (TIRF) microscope was used to assess the subcellular localization of Cx43 proteins. Dye injections were used to gain insight into the effects of propafenone on Cx43 metabolic coupling.

Results and Conclusion
Full length Cx43 protein levels were increased after treatment with propafenone and IF microscopy showed an intracellular accumulation of Cx43 protein, both on ectopically and endogenously expressed Cx43. Propafenone did not alter Cx43 half-life, in contrast to the lysosomal inhibitor chloroquine. Finally, gap-junctional coupling was decreased by chronic propafenone treatment. We conclude that the class 1c antiarrhythmic drug propanone increases Cx43 protein expression, resulting in its intracellular accumulation, as a side effect.
2-Aminoethoxydiphenyl borate prolongs atrial and shortens ventricular action potential duration through modulation of ion channels in the sarcolemma
Authors: Rivaud M , Bosada F M , Van Amersfoort S C M , Schumacher C A , Al Shama R F , De Groot J R , Verkerk A O , Coronel R F , Veldkamp M W 
Published: May 2024

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): PPP allowance, Health holland ZonmW

Background
2-Aminoethoxydiphenyl Borate (2-APB) has been shown to prevent atrial fibrillation in animal models, supposedly through its inhibitory action on the intracellular IP3 receptor and subsequent calcium regulation. However, 2-APB may modulate other ion channels or transporters and the cellular mechanisms associated with the prevention of atrial fibrillation are not well established. We aimed at unravelling the cellular effects of 2-APB on atrial and ventricular action potential and currents.

Methods
Rabbit (New Zealand White) left atrial and ventricular cardiomyocytes were isolated from Langendorff-perfused hearts by enzymatic dissociation. Adult human atrial cardiomyocytes were obtained by enzymatic dissociation from left atrial appendages (LAA) routinely removed from AF patients undergoing minimally invasive surgical pulmonary vein isolation. Action potentials (APs) and K+ currents were recorded at 36.5 °C with the amphotericin-B-perforated or whole cell patch clamp technique. RNA was isolated from left atrial and ventricular tissue and sequenced on an Illumina HiSeq4000 platform.

Results
In rabbit atrial cardiomyocytes, extracellular application of 5 uM 2-APB had no effect on action potential duration (APD), but extracellular application of 50 uM 2-APB prolonged the APD by 40% and inhibited an outward K+-current. Inversely, extracellular application of 5 uM 2-APB shortened the APD of rabbit left ventricular cardiomyocytes by 20% while extracellular application 50 uM 2-APB did not prolong the APD. Additionally, 2-APB did not modulate K+ currents in ventricular rabbit cardiomyocytes. Intracellular application (in the patch pipet) of 50 uM 2-APB did not have any effects on atrial APD, suggesting that IP3 receptor inhibition is not involved. However, subsequent application of 50 uM 2-APB in the bath prolonged the APD by 40% indicating that 2-APB prolongs atrial APD by inhibiting one or more K+ channel types in the sarcolemmal membrane from the extracellular site. RNA sequencing analysis revealed potential targets channel including TRP and Kv channels differentially expressed in atria versus ventricle. In human atrial cardiomyocytes from AF patients, application of 50 uM 2-APB prolonged APD by 25%.

Conclusions
2-APB prolongs rabbit and human atrial APD and shortens rabbit ventricular APD in a concentration dependent manner, most likely due to modulation of sarcolemmal K+ channels, differentially expressed in the two cell types. Pharmacological studies with 2-APB may reveal novel cellular mechanisms identifying potential new targets for the treatment of atrial fibrillation.
Targeting glutaminolysis pathway in trained immunity induced by oxidized low-density lipoprotein
Authors: Scarpa A , Groh L , Placek K , Matzaraki V , Bekkering S , Hiller K , Netea M G , Riksen N P 
Published: May 2024

Abstract
Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Deutsche Forschungsgemeinschaft DFG
Atherosclerosis is a chronic inflammatory disease of the arterial wall that causes cardiovascular disease. Monocyte-derived macrophages importantly contribute to atherogenesis. Monocytes can be primed to exhibit increased responsiveness upon a secondary non-related stimulus, a phenomenon called trained immunity. Intracellular metabolic and epigenetic reprogramming lays the basis of trained immunity. One of the stimuli that can induce trained immunity is oxidized low-density lipoprotein (oxLDL). oxLDL-trained monocytes exhibit an increase in metabolic pathways such as glycolysis, OXPHOS, and cholesterol biosynthesis.
The glutaminolysis pathway that fuels the tricarboxylic acid (TCA) cycle is essential for trained immunity induction by b-glucan, the cell wall component of Candida albicans.
The goal of this project is to investigate the involvement of the glutaminolysis pathway in oxLDL-induced trained immunity in isolated human primary monocytes. We exposed monocytes for 24 hours to oxLDL, allowed subsequent differentiation into macrophages for six days in culture medium, and assessed cytokine production capacity upon restimulation. Co-incubation of oxLDL with the inhibitor that blocks glutaminase, the enzyme that generates glutamate from glutamine, significantly reduced cytokine production capacity in oxLDL-trained cells. We further showed that glutaminolysis is necessary for oxLDL-induced increase in oxygen consumption (OCR) and glycolysis rate (ECAR). Single nucleotide polymorphisms in glutaminolysis genes were shown to correlate with ex vivo oxLDL-induced trained immunity in 243 healthy volunteers. These data suggest an important role of glutaminolysis in the establishment of oxLDL-induced trained immunity. To unravel the molecular mechanisms behind this observation we plan to perform epigenetic as well as metabolic analyses of the monocytes trained in the presence of the inhibitor.
Exploring the interactions between cardiomyocytes and endothelial cells to induce cardiac revascularization and regeneration
Authors: Vuerich R , Colliva A , Vodret S , Volpe M C , Braga L , Canarutto G , Piazza S , Pedrazzini T , Chiesa M , Cauteruccio M , Ramadan M , Zentilin L , Giacca M , Zacchigna S 
Published: May 2024

Abstract

Funding Acknowledgements
None.

Background
The adult heart is unable to regenerate after ischemic injury, primarily due to the incapacity of cardiomyocyte (CM) to re-enter in the cell cycle and endothelial cells (EC) to form new vessels. Interestingly, our recent data have unveiled that the decline of CM proliferation after birth is paralleled by a reduction in angiogenic potential. Nevertheless, the precise mechanisms governing angiogenesis inhibition in the adult heart remain elusive. We hypothesize that the terminal differentiation of CMs after birth leads to the paracrine suppression of EC proliferation.

Purpose
This study aims to identify membrane or secreted factors produced by adult CMs that inhibit EC proliferation and vessel formation in order to create new therapeutic strategies to induce cardiac revascularization and regeneration.

Material and Methods
To validate our hypothesis, ECs were co-cultured with CMs at distinct differentiation stages. Leveraging existing RNA sequencing datasets, we generated an interactome between ligands produced by CMs and receptors expressed by cardiac ECs. Through gain and loss of function studies, we identified pivotal anti-angiogenic factors and subsequently knocked-out the corresponding genes in Cas9 mice, with the aim of rescue the angiogenic potential of the adult heart. Concurrently, we assessed the ability of miR199a, a pro-proliferative miRNA known for inducing CM de-differentiation, to elicit EC proliferation and angiogenesis when delivered to the adult heart together with VEGF-A, utilizing adeno-associated viral vectors.

Results
Our findings revealed that adult CMs impaired EC proliferation. Comprehensive analysis identified 20 membrane and secreted factors, establishing 118 ligand-receptor interactions between CMs and ECs. Notably, KCNJ11 and CD74 emerged as key factors, as validated through gain and loss of function experiments. In vivo knockout of these genes in CMs led to significant increase in EC proliferation and the formation of neo-vessels. Furthermore, CM de-differentiation induced by miR199a demonstrated the capacity to promote EC proliferation and neo-vessel formation upon VEGF-A stimulation.

Conclusions
Collectively, this work demonstrated that fully mature CMs have a role in limiting angiogenic potential of the adult heart, likely explaining the failure of therapeutic angiogenesis approaches attempted so far. Our results suggest that innovative strategies targeting CM-EC cross-talk or inducing partial CM de-differentiation may open new therapeutic avenues in the field of cardiac regeneration.Adult CMs inhibit EC proliferationCMdedifferentiation rescues angiogenesis
Ketone-based metabolism and signalling in heart failure with preserved ejection fraction (HFpEF)
Authors: Wang R , Capone F , Capone F , Luo L , L L , Keller D , Keller D , Jung S , Jung S , Pauline F , Pauline F , Mertins P , Mertins P , Sotomayor-Flores C , Sotomayor-Flores C , Bode D , Bode D , Grunert N , Grunert N , Schiattarella G , Schiattarella G 
Published: May 2024

Abstract
Funding Acknowledgements
Type of funding sources: Public grant(s) – EU funding. Main funding source(s): DZHK
Deutsche Forschungsgemeinschaft (DFG-SFB_1470_A02)
Cardiac metabolic remodelling in heart failure with preserved ejection fraction (HFpEF) underlies the pathogenesis of this syndrome. Yet, our understanding of metabolic alterations occurring in HFpEF hearts is still limited. Ketone bodies (KBs) – β-hydroxybutyrate (β-OHB) in particular – can become a major, alternative source of fuel for failing hearts. However, the biological relevance of KBs goes beyond their role as energy carrier.
We used a ‘two-hit’ mouse model of HFpEF induced by high-fat-diet feeding and L-Name, aiming to evaluate metabolic alteration among various energy substrates (e.g., glucose, fatty acids), in particular the role of β-OHB in HFpEF hearts. Here, we showed that HFpEF heart exhibited impaired glucose oxidation and increased fatty acid oxidation. Impaired glucose oxidation was attributed to the reduced pyruvate dehydrogenase (PDH) activity and increased phospho-PDH expression via the activation of PDH kinase 4 (PDK4). The oxidation of β-OHB was reduced as a result of decreased β-OHB dehydrogenase (BDH1) and succinyl-coA-3-oxaloacid CoA transferase (SCOT) expression. Furthermore, we showed that β-OHB governs mitochondrial function and myocardial bioenergetics in HFpEF, acting as protein modifiers through β-hydroxybutyrylation (Kbhb) – a lysine-based post-translational modification (PTM). Immunoblot analysis for pan- β-hydroxybutyryllysine revealed a significant increase in Kbhb levels in the heart of ‘two-hit’ HFpEF mice compared to control mice. To identify the specific proteins and sites of Kbhb in the heart, Kbhb peptides from control and HFpEF mouse hearts were isolated by immunoaffinity enrichment using a pan-Kbhb antibody-conjugated beads. Volcano plot and gene ontology analysis showed that the most Kbhb-modified proteins in HFpEF hearts are mitochondrial enzymes. Glutamate oxaloacetate transaminase (GOT2) is one of the top hits, participating in the TCA cycle and malate-aspartate shuttle (MAS) in the mitochondria. We further demonstrated that GOT2 activity and MAS were enhacned in the mitochondrial of HFpEF hearts.
To conclude, in the heart of ‘two-hit’ model of HFpEF, the oxidation of pyruvate was impaired resulting from decreased PDH activity. The oxidation of β-OHB was also reduced due to the down-regulated BDH1 activity. Instead of oxidation, Kbhb was highlighted as the primary mode of KB utilisation in HFpEF, affecting the GOT2 enzyme activity in MAS pathways. Our results shed light on identifying a novel role of KBs, which can facilitate the understanding of pathogenesis in HFpEF.Ketone-based metabolism and signalling