bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

Abstract

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&lt;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

Abstract

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

Abstract
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

Abstract

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&lt;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

Abstract

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&lt;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&lt;0,01). Proliferation in HVSMC increased significantly (p&lt;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&lt;0,05), KLF4 (p&lt;0,05), CD69 (p&lt;0,01), MAP2K4 (p&lt;0,05) and BCL211 (p&lt;0,01). Additionally, CD69 and KLF4 expression increased significantly either in HVECs (p&lt;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

Abstract

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&lt;0,01) and increased migratory function (+40%,p&lt;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&lt;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&lt;0,01) and reduced energy charge (-40%,p&lt;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

Abstract

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 &lt; .05). None of the remaining parameters revealed any difference for MRONJ-affected bone in CBCT (P &gt; .05). FD, lacunarity, MGV, BA/TA, and Tb.Sp could distinguish MRONJ-affected trabecular bone in PR (P &lt; .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

Abstract
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

Abstract
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

Abstract

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.