bluuebunny/crossref_metadata_2025 huggingface dataset:
Granuloma annulare and the risk of autoimmune conditions: a multicentre retrospective cohort analysis
Authors: Garate David , Thang Christopher J , Schmidt Madelyn , Ravanassa Shayan , Jimenez Antonio R , Ross Lindy S , Golovko George , Barbieri John S , Wilkerson Michael G
Published: May 2024

Prior studies and case reports have postulated potential associations between granuloma annulare (GA) and various autoimmune conditions, but these relationships remain unclear. Using the TriNetX Research Network, we found that GA was significantly associated with an increased risk of systemic lupus erythematosus, rheumatoid arthritis, autoimmune thyroiditis and type 1 diabetes mellitus, along with point estimates suggesting an increased risk of systemic sclerosis and uveitis. Overall, our study uses epidemiological data to provide valuable evidence supporting an increased risk of autoimmune disorders in people with GA.
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

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

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

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

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

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

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
Mutation in PDGFRbeta: a potential new pathogenic variant for acquired mitral valve prolapse
Authors: Wu H W , El Ouraoui M , Van Wijnwaarden A L , Mousavi Gourabi B , Smits A M , Barge-Schaapveld D Q C M , De Vries A A F , Jukema J W , Kruithof B P T , Ajmone Marsan N
Published: May 2024

Funding Acknowledgements

None.

Background

Mitral valve prolapse (MVP) due to myxomatous degeneration is characterized by familial clustering, but few pathogenic genes have been identified. In a genetic screening program for MVP patients, we identified a platelet-derived growth factor receptor β (PDGFRβ)-E162K missense variant in a family with a bi-leaflet MVP phenotype in four affected family members. PDGFRβ is a receptor tyrosine kinase, which plays an important role in vascular development.

Purpose

In this study, we investigated whether the E162K substitution in PDGFRβ could lead to mitral valve abnormalities.

Methods

To assess the functional effects of the missense variant, proliferation and migration assays were performed using HeLa cells overexpressing wild-type (WT) or mutant PDGFRβ. Immunostainings were performed to determine PDGFRβ expression in the diseased valves from a family member undergoing surgical operation and from additional patients operated for MVP. Mouse models were created with the mouse equivalent of the human PDGFRβ-E162K variant (PDGFRβ-E161K) using CRISPR/CAS9. The variant was not lethal and transthoracic echocardiography was performed on adult homozygous PDGFRβE161K/E161K mice, heterozygous PDGFRβE161K/- mice, and WT mice . In addition, histology was performed in adult and neonatal mice to detect potential valvular defects.

Results

HeLa cells expressing PDGFRβ-E162K and stimulated with PDGFD showed reduced proliferation and migration, suggesting a loss of function of the mutant receptor. PDGFRβ was expressed in human diseased mitral valves. Echocardiographic evaluation revealed a significant larger mitral valve annulus diameter in both PDGFRβE161K/- and PDGFRβE161K/E161K compared to WT (WT: 1.97 mm; PDGFRβE161K/-:2.13, p=0.031; PDGFRβE161K/E161K:2.42 mm, p<0.0001). Histological data in PDGFRβE161K/E161K mice showed significant leaflet thickening in the free edge (FE) of the posterior mitral valve leaflet (PMVL) compared to WT (WT: 72.34 ɥm; PDGFRβE161K/E161K: 109.6 ɥm, p=0.040) and a larger leaflet area of the PMVL compared to WT (WT: 25280 ɥm²; PDGFRβE161K/E161K: 44494 ɥm², p=0.020). Moreover, an altered organization of the extracellular matrix reminiscent of the myxomatous degeneration in MVP was observed. These alterations were not observed in mutant neonatal hearts.

Conclusions

The PDGFRβ-E162K variant is associated with familial MVP and alters the function of PDGFRβ. Mice harboring this mutation display acquired mitral valve defects.
Nuclear eNOS interacts with and S-nitrosates ADAR1 to modulate endothelial gene expression
Authors: Zhou X , Guenther S , Kuenne K , Wittig I , Fleming I , Siragusa M
Published: May 2024

Funding Acknowledgements

Type of funding sources: Foundation. Main funding source(s): Collaborative Research Centre (CRC 1366); Chinese Scholarship Council (CSC)

Introduction and Aim

Nitric oxide (NO) generated by the endothelial NO synthase (eNOS) regulates vascular tone and endothelial homeostasis to counteract vascular inflammation. The effects of NO are attributable to its interaction with heme-containing proteins e.g., soluble guanylyl cyclase, or to the S-nitrosation of proteins. Most eNOS is localized to the cell membrane or the Golgi apparatus, however, it has also been detected in the nucleus. In this study, we assessed the role of nuclear eNOS in endothelial cells.

Methods and Results

Confocal microscopy studies confirmed the presence of eNOS in the nucleus of unstimulated human and murine endothelial cells (ex vivo and in vitro) and stimulation with vascular endothelial growth factor (VEGF) increased eNOS nuclear translocation. Co-immunoprecipitation studies coupled with proteomics revealed the VEGF-dependent association of nuclear eNOS with 81 proteins involved in RNA binding and processing. Among these, nuclear eNOS bound double-stranded RNA-specific adenosine deaminase 1 (ADAR1), an enzyme involved in RNA editing via the deamination of adenosine (A) to inosine (I) in double-stranded RNA (dsRNA). ADAR1 was S-nitrosated in human endothelial cells and the knockdown of eNOS was associated with an increase in dsRNA (immunofluorescence) and altered ADAR1-mediated A-I editing (RNA seq). The accumulation of dsRNA in human endothelial cells lacking eNOS led to the activation of the interferon α/β signalling pathway (e.g., MX2, OASL, OAS1/2/3, RNASEL) and the downregulation of cell cycle-related genes. As a result, growth factor-stimulated cell proliferation was abrogated and basal as well as tumour necrosis factor- or H2O2-induced cell death were increased. Furthermore, the increased dsRNA in endothelial cells lacking eNOS elicited the activation of interferon-induced, dsRNA-activated protein kinase, which in turn phosphorylated the eukaryotic translation initiation factor 2A. Accordingly, new protein synthesis was inhibited following eNOS knockdown (immunofluorescence and mass spectrometry).

Conclusion

These results demonstrated that eNOS/NO signalling regulates endothelial type I interferon signalling and affects translation, cell cycle and cell death through S-nitrosation of ADAR1.
Human atherosclerotic plaque macrophages associate with clinical presentation and follow diverse lineage differentiation routes
Authors: Prange K , Bel-Bordes G , Depuydt M A C , Kempkes R W M , De Jong M D M , Mol B M , Slutter B , Bot I , Neele A E , De Kleijn D P V , De Borst G J , Kuiper J , Pasterkamp G , Mokry M , De Winther M P J
Published: May 2024

Funding Acknowledgements 

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Dutch Research Council Veni grant. Netherlands Heart Foundation

Atherosclerosis is characterized by inflammation and lipid accumulation, leading to complications such as stroke and myocardial infarction. Various macrophage subsets are present in human atherosclerotic plaques. However, it is unclear how subsets are interrelated and contribute to the disease process and its clinical outcomes.

Here, we employed single-cell RNA-seq (scRNA-seq) on blood and plaque material from carotid endarterectomy patients enrolled in the AtheroExpress (AE) cohort to define regulatory pathways of macrophage recruitment, differentiation, and development. Additionally, we used these scRNA-seq data to deconvolute a large AE bulk RNA-seq cohort to allow correlating cellular subsets in plaques with clinical traits.

Interestingly, we found that macrophages and their subsets are the only cell population significantly associated with major adverse cardiac events (MACE) during a 3-year follow-up period. Subsequently, we could divide plaque macrophages into 3 populations: inflammatory, tissue-resident / lipid-associated (TREM2 high), and foamy macrophages (TREM1 high). While the latter two are correlated with MACE, the inflammatory macrophages are inversely correlated with statin use.

Cellular fate analyses revealed two major paths of macrophage development in the plaque. First, non-classical monocytes entering the plaque, turning into inflammatory cells before becoming foamy and eventually dying. Second, resident-like macrophages that turn into either inflammatory- or lipid-associated macrophages, before meeting their foamy demise.

Together, these findings provide important insights on macrophages in atherosclerosis and suggest new leads to discover therapeutic targets for this disease.
miRNA signature related to atherosclerotic lesion induced shear stress modifications in familial hypercholesterolemia patients with subclinical atherosclerosis: a bioinformatics systems biology study
Authors: Padro T , Escate R , Perez De Isla L , Muniz-Grijalbo O , Mata P , Badimon L
Published: May 2024

Funding Acknowledgements

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Institute of Health Carlos III (ISCIII)

Background

Patients with familial hypercholesterolemia (FH), even when appropriately treated, are at risk of premature clinical atherosclerotic cardiovascular disease (ASCVD) presentation. Although atherosclerosis (AT) is directly related to hemodynamic disturbances affecting the vascular geometry, there is scarce information on whether there is miRNA-mediated/flow-mediated modulation of atherosclerosis in FH.

Purpose

To identify a differential plasma miRNA signature related to fluid shear stress (FSS) in well controlled FH-patients with asymptomatic atherosclerosis.

Methods

FH-patients (N=49) with subclinical atherosclerosis, identified by computed tomographic angiography, and a reference cohort of non-FH subjects (N=32), both from a national FH-Registry, were included in the study. Coronary plaque volume and phenotype was analyzed by means of the QAngio CT imaging system. miRNAs associated to FSS were defined in silico by bioinformatics (KEGG, Cytoscape, GTEX portal, miRWalk, miRBase tools) and validated in plasma samples by RT-qPCR using TaqMan technology.

Results

By in silico analysis, 7 miRNAs targeting genes involved in features of functions (ROS production, oxidative stress, leukocyte transendothelial/migration-inflammation and vasodilation) of shear stress-induced cellular dysfunction and atherosclerosis (KEGG, hsa05418) were identified by miRWalk and miRBase databases. Of the 7 miRNAs, a 6-miRNA signature showed significantly lower plasma levels in FH-patients than in the non-FH group (P<0.01 for: miR-30e-5p, miR-378a-3p, let-7d-3p, and miR-550-5p; P=0.03 for miR-324-3p). In the FH-group, miR-324-3p, miR-30e-5p, miR-378a-3p and let-7d-3p significantly discriminated patients stratified by the plaque volume of non-calcified lesions (fibro-lipidic phenotype), using the median plaque volume as cut-off value. And even further, a high percentage (65-70%) of FH-subjects with low levels of the 4 miRNA had plaque volumes above the median. Interestingly, this miRNA-signature did not differentiate FH-patients stratified by the volume of calcified plaques.

Conclusion

FH-patients show a significantly lower content of an atherosclerosis/shear stress signature of 4 miRNAs targeting genes involved in subclinical atherosclerosis progression in FH.
MiR-32-5p as potential key mediator of vascular remodeling enhances cell proliferation of vascular smooth muscle cells
Authors: Wils T D , Kalies K , Guerlach S , Koester J M , Boettcher S , Daniel F , Hehl L , Knoepp K , Dutzmann J , Sedding D G
Published: May 2024

Funding Acknowledgements

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): University Hospital Halle, Martin-Luther-University Halle-Wittenberg

Background and purpose

The dysfunctionality of endothelial cells (EC) and smooth muscle cells (SMC) contributes to the progression of vascular remodeling and subsequent cardiovascular pathologies. Several microRNAs (miRNA) have been revealed as key regulators of EC and SMC function, therefore suggesting them as potential targets in the modulation of vascular remodeling. Here, we investigate the potential role of miR-32-5p in vascular cell function and its potential as a therapeutic target in vascular remodeling.

Material and Methods

Initial screenings by qRT-PCR analysis for miR-32-5p expression were performed in murine femoral artery tissue on day 10 and 21 after wire-induced injury in C57Bl6 mice, serving as restenosis model. MiR-32-5p expression was assessed using qRT-PCR in human vascular endothelial (HVECs) and human vascular smooth muscle cells (HVSMC). The role of miR-32-5p on cellular functions, as migration and proliferation, was determined by performing scratch wound and BrdU assays. Potential miR-32-5p downstream targets were identified from literature and their expression levels upon interference with miR-32-5p evaluated on mRNA and protein levels.

Results

MiR-32-5p expression was decreased over time in the wire-induced injury model. In vitro, miR-32-5p expression was significantly decreased in replicative senescent HVSMC and HVEC (p<0,05) and relatively higher in HVSMC than in HVEC for all conditions. Under growth conditions an elevated expression of miR-32-5p in HVEC and significantly increased expression in HVSMC was detected (p<0,01). Proliferation in HVSMC increased significantly (p<0,05) by pre-miR-32-5p transfection, but remained unaffected in HVEC. Migration of HVECs and HVSMCs was improved by miR-32-5p and decelerated after antagomiR interference. Systemic literature target research revealed several potential targets in both cell types, of which the following were significantly downregulated in HVSMC after pre-miR transfection including PIK3r3 (p<0,05), KLF4 (p<0,05), CD69 (p<0,01), MAP2K4 (p<0,05) and BCL211 (p<0,01). Additionally, CD69 and KLF4 expression increased significantly either in HVECs (p<0,05).

Conclusion

Our results suggest that miR-32-5p may be a substantial regulator in HVEC and HVSMC senescence and function and subsequently in vascular remodeling. To fully evaluate its role in the pathogenesis of atherosclerosis and therefore, its therapeutic potential in cardiovascular diseases, further investigations are needed. Additional experiments with the purpose of investigating miR-32-5p-target interactions and apoptotic effects will be performed shortly.
The sterol element binding protein 1c (SREBP1c) preserves cellular metabolism and immunosuppressive function of regulatory t cells
Authors: Bonacina F , Procaccini C , Pedretti S , Stucchi G , Moregola A , Bogie J , Matarese G , Mitro N , Norata G D
Published: May 2024

Funding Acknowledgements

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Cariplo Foundation

PRIN (Italian Ministry of University and Research)

Aim

Balance of cellular metabolism is increasingly recognized as a hallmark of physiologic cellular functions; this also extends to cells of the immune system. Here, we aimed at investigating the role of SREBP1c, a key protein regulating intracellular fatty acid (FA) metabolism, on the metabolism and function of Treg, that play a key role in immune tolerance maintenance.

Material and Methods

A detailed immunophenotyping through flow cytometry and metabolic profiling (metabolomics and seahorse analysis) of isolated Tregulatory (CD4+CD25+, nTreg) and in vitro induced Treg (iTreg) cells were performed together with in vitro and in vivo assays of Treg function from SREBP1c KO and WT littermates. RNAseq and lipidomics was performed on iTreg.

Results

Srebp1c deficiency reduced suppressive (-21%,p<0,01) and increased migratory function (+40%,p<0,05) of nTreg and iTreg, tested by in vitro assays. In vivo, Srebp1c KO mice presented reduced circulating and tissues’ level of Treg compared to WT mice (-66%,p<0,01) and nTreg from Srebp1c KO mice worsened Experimental Autoimmune Encephalomyelitis progression compared to WT nTreg. We addressed impaired Treg function to metabolic alteration due to Srebp1c deficiency: KO iTreg showed an increased glycolytic potential with preserved mitochondrial function coupled to accumulation of glycolytic metabolites and lactate (+20%,p<0,01) and reduced energy charge (-40%,p<0.01). We associated the switch to anaerobic glycolysis in KO Treg to an impaired lipid metabolism, as suggested by impaired abundance of lipid species (TG, LPC and PC); this was confirmed by RNAseq showing a downregulation of lipid metabolism’s pathways and upregulation of glycolysis. In parallel, tolerogenic response was impaired, a phenotype associated to a reduced transcription and activation of Foxp3, the master regulator of Treg function.

Conclusion

By restrain glycolysis and preserving lipid metabolism, our data have identified SREBP1c as a checkpoint crucial for the immunometabolic suppressive response of Tregs.
Comparison of quantitative radiomorphometric predictors of healthy and MRONJ-affected bone using panoramic radiography and cone-beam CT
Authors: Aslan Elif , Onem Erinc , Mert Ali , Baksi B Guniz
Published: May 2024

Objectives

To determine the most distinctive quantitative radiomorphometric parameter(s) for the detection of MRONJ-affected bone changes in panoramic radiography (PR) and cone-beam CT (CBCT).

Methods

PR and sagittal CBCT slices of 24 MRONJ patients and 22 healthy controls were used for the measurements of mandibular cortical thickness (MCT), fractal dimension (FD), lacunarity, mean gray value (MGV), bone area fraction (BA/TA), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), trabecular number (Tb.N). MCT was measured in the mental foramen region. While FD and lacunarity were measured on mandibular trabecular and cortical regions-of-interest (ROIs), the remaining parameters were measured on trabecular ROIs. The independent samples t-test was used to compare the measurements between the MRONJ and control groups for both imaging modalities (P = .05).

Results

MCT was the only parameter that differentiated MRONJ-affected bone in both PR and CBCT (P < .05). None of the remaining parameters revealed any difference for MRONJ-affected bone in CBCT (P > .05). FD, lacunarity, MGV, BA/TA, and Tb.Sp could distinguish MRONJ-affected trabecular bone in PR (P < .05). The correspondent ROI for both imaging methods that was reliable for detecting MRONJ-affected bone was the trabecular bone distal to the mental foramen above the inferior alveolar canal (ROI-3).

Conclusions

MCT is a reliable parameter for the discrimination of MRONJ-affected bone in both PR and CBCT images. PR may be used to detect MRONJ-affected trabecular bone using FD, lacunarity, MGV, BA/TA, and Tb.Sp measurements as well.
High-integrity Pueraria montana var. lobata genome and population analysis revealed the genetic diversity of Pueraria genus
Authors: Huang Xuan-Zhao , Gong Shao-Da , Shang Xiao-hong , Gao Min , Zhao Bo-Yuan , Xiao Liang , Shi Ping-li , Zeng Wen-dan , Cao Sheng , Wu Zheng-dan , Song Jia-Ming , Chen Ling-Ling , Yan Hua-bing
Published: May 2024

Pueraria montana var. lobata (P. lobata) is a traditional medicinal plant belonging to the Pueraria genus of Fabaceae family. Pueraria montana var. thomsonii (P. thomsonii) and Pueraria montana var. montana (P. montana) are its related species. However, evolutionary history of the Pueraria genus is still largely unknown. Here, a high-integrity, chromosome-level genome of P. lobata and an improved genome of P. thomsonii were reported. It found evidence for an ancient whole-genome triplication and a recent whole-genome duplication shared with Fabaceae in three Pueraria species. Population genomics of 121 Pueraria accessions demonstrated that P. lobata populations had substantially higher genetic diversity, and P. thomsonii was probably derived from P. lobata by domestication as a subspecies. Selection sweep analysis identified candidate genes in P. thomsonii populations associated with the synthesis of auxin and gibberellin, which potentially play a role in the expansion and starch accumulation of tubers in P. thomsonii. Overall, the findings provide new insights into the evolutionary and domestication history of the Pueraria genome and offer a valuable genomic resource for the genetic improvement of these species.
Characterization of transcriptome changes in saline stress adaptation on Leuciscus merzbacheri using PacBio Iso-Seq and RNA-Seq
Authors: Yu Dan , Zhou Min , Chen Wenjun , Ding Zufa , Wang Cheng , Qian Yuting , Liu Yang , He Shunping , Yang Liandong
Published: May 2024

Leuciscus merzbacheri is a native fish species found exclusively in the Junggar Basin in Xinjiang. It exhibits remarkable adaptability, thriving in varying water conditions such as the saline waters, the semi-saline water, and the freshwater. Despite its significant economic and ecological value, the underlying mechanisms of its remarkable salinity tolerance remain elusive. Our study marks the first time the full-length transcriptome of L. merzbacheri has been reported, utilizing RNA-Seq and PacBio Iso-Seq technologies. We found that the average length of the full-length transcriptome is 1,780 bp, with an N50 length of 2,358 bp. We collected RNA-Seq data from gill, liver, and kidney tissues of L. merzbacheri from both saline water and freshwater environments and conducted comparative analyses across these tissues. Further analysis revealed significant enrichment in several key functional gene categories and signalling pathways related to stress response and environmental adaptation. The findings provide a valuable genetic resource for further investigation into saline-responsive candidate genes, which will deepen our understanding of teleost adaptation to extreme environmental stress. This knowledge is crucial for the future breeding and conservation of native fish species.
Masquerade of an emergency: cardiac tamponade as a deceptive presentation of primary cardiac diffuse large b-cell lymphoma—a case report
Authors: Heeringa Tijn J P , Roscam Abbing Reinout L P , van Leeuwen Gijs A M , van Putte Bart P , de Bruin Anthonius F J
Published: May 2024

Background

Primary cardiac diffuse large B-cell lymphoma (CDLBCL) is an exceptionally rare entity, estimated to represent less than 1% of all primary cardiac tumours. In this case report, we emphasize the diagnostic importance of multimodality imaging and the need for additional procedures, such as tissue biopsy, in a case with a primary cardiac lymphoma presenting with cardiac tamponade.

Case summary

An 80-year-old male was admitted to the emergency department with a life-threatening tamponade demanding immediate sternotomy. Pre-operative echocardiography unveiled pericardial effusion and a thickened apex. While computed tomography ruled out an aortic dissection, surgery revealed an unexpected vascular-rich mass at the right ventricle and apex, too perilous for biopsy. Post-operative imaging misinterpreted this mass as a benign haematoma. Subsequently, the patient was admitted to the intensive care unit, but after a conservative treatment strategy, the patient died. An autopsy revealed a primary CDLBCL.

Discussion

This case demonstrates the deceptive nature of primary CDLBCL, often complicated by cardiac tamponade. It underscores the pivotal role of pathologic assessment, even amidst the perils of sternotomy, to determine the origin of abnormal cardiac masses. A heightened awareness among physicians is imperative, for such elusive diagnoses may slip by, with potentially fatal outcomes.
Severe tricuspid regurgitation with biannular disjunction requiring surgical treatment: a case report
Authors: Masuda Mayuka , Imanishi Junichi , Inoue Takeshi , Okuda Masanori
Published: May 2024

Background

Tricuspid annular disjunction (TAD) is an annular disjunction of the right-sided heart. Although TAD is often concomitant with mitral annular disjunction (MAD), it often presents as mitral regurgitation (MR), rather than tricuspid regurgitation (TR). While the clinical significance of MAD has been well-established, there is still little data on TAD. This is a rare case of severe TR due to TAD that appears to be isolated from MAD.

Case summary

A 63-year-old female complaining of pre-syncope and dyspnoea on exertion was referred to our department. Initial transthoracic echocardiography showed MR and TR due to tricuspid valve prolapse (TVP). On transoesophageal echocardiography, the TVP consisted of an excessively redundant anterior leaflet, where the annular disjunction and severe regurgitation were formed. She recently underwent mitral and tricuspid valve plasties for symptomatic primary severe TR.

Discussion

This case report emphasizes the clinical significance of TAD as a potential cause of severe TR, even without significant MR. Tricuspid annular disjunction progresses more gradually compared with MAD. This case suggests that remodelling of the right atrium, particularly in chronic atrial fibrillation, may contribute to the development of TR. Despite diagnostic challenges due to the flexible and dynamic nature of the tricuspid annulus, this is the first report of TAD-induced severe TR necessitating surgical intervention. Accurately diagnosing TAD remains challenging with current imaging modalities, emphasizing the need for improved diagnostic tools to optimize treatment strategies.
Case report of a double-wave re-entry atrial flutter in a patient with atrial cardiomyopathy
Authors: Jacinto Sofia , Portugal Guilherme , Valente Bruno , Cunha Pedro , Oliveira Mário
Published: May 2024

Background

Double-wave macrore-entry is a rare mechanism of atrial tachycardia with limited documentation in the literature. We present a three-dimensional documentation of a double-wave ‘typical’ atrial flutter in a patient with extensive atrial cardiomyopathy.

Case summary

A 78-year-old female with a history of atrial cardiomyopathy and dual-chamber pacemaker for sinus node disease presented with palpitations and incessant atrial flutter. Electrophysiological study revealed a regular tachycardia with a cycle length (TCL) of 230 ms, with proximal to distal coronary sinus (CS) activation. Three-dimensional mapping identified two independent wavefronts circulating the cavotricuspid isthmus (CTI), each with a TCL of 460 ms. Cavotricuspid isthmus ablation resulted in conversion into a distinct tachycardia with left atrial roof origin. Linear ablation in this location slowed the TCL to 435 ms with concentric CS activation and another CTI dependent atrial flutter was mapped, this time with only one wavefront of activation. Further ablation with a second, more lateral, line in the CTI led to tachycardia interruption. Given the extensive atrial scarring and high arrhythmic recurrence risk, atrioventricular node ablation was performed.

Discussion

Double-wave re-entrant tachycardias were primarily observed in experimental models, precipitating acceleration of ventricular and supraventricular tachycardias via extrastimulation. In our case, there is documentation of a spontaneous double-wave of activation around the CTI, representing the first documented double-wave ‘typical’ atrial flutter. Unlike other cases in the literature, the two wavefronts were equidistant, which resulted in a regular tachycardia with TCL that was half of the single-wave cycle length. Three-dimensional propagation mapping was essential to visualize the two distinct wavefronts.
Economic Shocks and Populism
Authors: Panunzi Fausto , Pavoni Nicola , Tabellini Guido
Published: May 2024

We study how voters’ preferences between a safe incumbent and a risky opponent change in the aftermath of a negative aggregate shock. With reference-dependent preferences, economically disappointed voters become risk lovers, and are hence attracted by the more risky candidate. Survey data from the German Socio-Economic Panel are consistent with our assumptions and theoretical predictions on voters’ behaviour.
Methicillin-resistant Staphylococcus aureus and coagulase-negative Staphylococcus produce antimicrobial substances against members of the skin microbiota in children with atopic dermatitis
Authors: Guimarães Lorrayne Cardoso , Garcia Gizele Duarte , Cavalcante Fernanda Sampaio , Dias Graciela Maria , de Farias Felipe Miceli , Saintive Simone , Abad Eliane de Dios , Ferreira Dennis de Carvalho , dos Santos Kátia Regina Netto
Published: May 2024

Coagulase-negative Staphylococcus (CoNS) species inhibiting Staphylococcus aureus has been described in the skin of atopic dermatitis (AD) patients. This study evaluated whether Staphylococcus spp. from the skin and nares of AD and non-AD children produced antimicrobial substances (AMS). AMS production was screened by an overlay method and tested against NaOH, proteases and 30 indicator strains. Clonality was assessed by pulsed-field gel electrophoresis. Proteinaceous AMS-producers were investigated for autoimmunity by the overlay method and presence of bacteriocin genes by polymerase chain reaction. Two AMS-producers had their genome screened for AMS genes. A methicillin-resistant S. aureus (MRSA) produced proteinaceous AMS that inhibited 51.7% of the staphylococcal indicator strains, and it was active against 60% of the colonies selected from the AD child where it was isolated. On the other hand, 57 (8.8%) CoNS from the nares and skin of AD and non-AD children, most of them S. epidermidis (45.6%), reduced the growth of S. aureus and other CoNS species. Bacteriocin-related genes were detected in the genomes of AMS-producers. AMS production by CoNS inhibited S. aureus and other skin microbiota species from children with AD. Furthermore, an MRSA colonizing a child with AD produced AMS, reinforcing its contribution to dysbiosis and disease severity.
(Re-) Defining “Successful Aging” as the Endpoint in Clinical Trials? Current Methods, Challenges, and Emerging Solutions
Authors: Schönstein Anton
Published: May 2024

Background and Objectives

“Successful aging” as defined in Rowe & Kahn’s model aims to include the major domains of aging under a single overarching concept. From here the question arises how “successful aging” itself can be implemented as an endpoint in clinical research in a way that it is compliant to methodological regulatory framework and recommendations as formulated, for example, by the FDA. This article discusses from an applied perspective approaches and examples of how “successful aging” as a multi domain concept can be put into measurement practice.

Research Design and Methods

Narrative literature review.

Results

Existing methods of merging insights from, e.g., functional, social, and cognitive outcomes, can fail to perform under conditions of extreme heterogeneity, as often present in samples of older adults. In research on rare diseases, the methodological approach of a Multi Domain Responder Index (MDRI) has been proposed to handle heterogeneity. MDRIs may also provide a solution to combine aging outcomes from different domains into a single “successful aging” endpoint. However, strict measurement criteria will need to be fulfilled to find acceptance in decision making and validated meaningful change thresholds are a critical prerequisite.

Discussion and Implications

Gerontology can spearhead methodological approaches of handling multidimensional measurement under heterogeneity by validating a “successful aging” MDRI suitable for use as an endpoint in clinical research.