bluuebunny/crossref_metadata_2025_split huggingface dataset:
Diagnostic ability of miR-19a and Let-7f along with platelet and leukocyte extracellular vesicles in acute ischemic stroke patients
Authors: Eyileten C , Nowak A , Shahzadi A , Jarosz-Popek J , Jakubik D , Gasecka A , Van Der Pol E , Wolska M , Mirowska-Guzel M , Czlonkowska A , Postula M 
Published: October 2022

Abstract

Background
Ischemic stroke (IS) is one of the most frequent causes of death. Since miRNAs have been illustrated to play an important role in various processes through regulation of multiple genes, and platelet function, their utility as novel biomarkers should be determined.

Purpose
We aimed to analyze the circulating platelet-derived miR-19a-3p, miR-186-5p, Let-7f, platelet-extracellular vesicles (EVs), leukocyte-EVs, and endothelial-EVs levels 24-h and 7-days after IS as novel diagnostic and prognostic/predictive biomarkers in 28 acute IS and 35 control patients.

Methods
Blood samples of 28 patients diagnosed with acute IS with hyper platelet reactivity were collected 24-h and 7-days after stroke and 35 age- and gender-matched individuals free of stroke with multiple risk factors for cardiovascular disease. Platelet reactivity was assessed by AA-, TRAP-, ADP-induced platelet aggregometry. PlasmaRNA was extracted; quality RNA was assessed: fluorometric assay; RT-PCR for miRNAs expression measurement and flow-cytometry for EVs determination p&lt;0.05.

Results
Patients with IS on day-1 had significantly higher platelet reactivity assessed by AA-induced platelet aggregometry compared to controls (p=0.001). Patients with normal platelet activation had significantly higher miR-186-5p expression levels compared to patients with HPR at day-1 acute-stroke (p=0.034). Seven days after acute stroke, expression levels of miR-186-5p significantly decreased in the same patients with normal platelet reactivity (p=0.036). Patients with HPR had significantly elevated platelet-EVs (CD62) concentration compared to patients with normal platelet reactivity at the day of 1 acute-stroke (p=0.012). Similarly, patients with HPR had significantly higher leukocyte-EVs (CD45) concentration compared to patients with normal platelet function at day-1 acute-stroke (p=0.002). Diagnostic values of baseline miRNAs and EVs were evaluated with receiver operating characteristic curve analysis. ROC curve showed that pooling the miR-19a-3p expressions, platelet-EVs, and leukocyte-EVs concentration yielded a higher AUC than the value of each individual biomarker as AUC was 0.893 (95% CI, 0.79–0.99). Patients with moderate stroke had significantly elevated miR-19a-3p expression levels compared with patients with minor stroke at the day of acute IS. AUC in ROC curve analysis was 0.867, (95% CI, 0.74–0.10) p=0.001.

Conclusions
Our analysis showed alteration of circulating miRNAs and EVs after IS. Combination of miR-19a-3p, Let-7f, platelet-EVs and leukocyte-EVs might have a diagnostic value for acute stroke and miR-19a-3p can predict the severity of stroke in IS patients.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): National Science Centre - Narodowe Centrum Nauki
Leptin mediates protective effects on the vasculature
Authors: Filipova V , Stuerzebecher P E , Kralisch S , Schubert M R , Hoffmann A , Oliveira F , Sheikh B , Blueher M , Kogel A , Scholz M , Miehle K , Ebert T , Laufs U , Toenjes A , Boeckel J N 
Published: October 2022

Abstract

Introduction and purpose
Lipodystrophy (LD) syndromes are characterized by the loss of adipose tissue resulting in metabolic complications and accelerated atherosclerosis. The systemic concentration of the adipokine leptin is reduced in LD as a result of adipose tissue deficiency. A therapeutical option to treat LD is the substitution of leptin, which improves metabolic complications and reduces mortality. However, the vascular effects of leptin remain largely unknown.
Here we analyze the direct effects of leptin on the vascular system and the development of atherosclerosis.

Methods and results
Treatment of human endothelial cells (ECs) with leptin reduced endothelial inflammation and the process of endothelial-to-mesenchymal transition (EndMT) (CNN1, −41.4%, p&lt;0.05, n=4). In addition, leptin administration prevented the EndMT-induced increase of endothelial permeability. The protective effect of leptin on EndMT was confirmed in vivo in a combined lipodystrophic and atherosclerosis-prone mouse model (LDLR−/−; aP2-nSrebp1c). Treatment of the mice with leptin (3.0 mg/kg body weight daily for 8 weeks) decreased EndMT. Leptin showed no effect on plaques size but reduced the protrusion of plaques in atherosclerotic areas of the aortic roots (−31%, p&lt;0.05, n=4–6).
Cytokine screening revealed an increase of the growth differentiation factor 15 (GDF15) in serum of LD patients (+26.2%, p&lt;0.05, n=53–58) and in ECs after EndMT (+138%, p&lt;0.05, n=6743–10920). This increase was reversed using leptin treatment in ECs undergoing EndMT, in the LD mice model, and in LD patients after 4 weeks of leptin administration. Indeed, treatment of endothelial cells with GDF15 induced EndMT (CNN1, +7.7-fold-control, p&lt;0.05, n=3), and impaired EC barrier function. Neutralizing antibodies targeting GDF15 inhibited EndMT-mediated expression of mesenchymal genes (CNN1, −54%, p&lt;0.05, n=4). The treatment of ECs with serum from LD patients induced EndMT and the increase of mesenchymal marker expression was inhibited with additional administration with neutralizing antibodies targeting GDF15 (CNN1, −28%, p&lt;0.05, n=3).

Conclusion
Our findings indicate that EndMT is part of the cardiovascular disease progression in lipodystrophy syndromes. Leptin treatment has direct protective vascular effects by preventing inflammation, EndMT, and maintaining endothelial integrity.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)
Left ventricular volumes on positron emission tomography: validation against cardiovascular magnetic resonance
Authors: Han Y , Ahmed A I , Saad J M , Malahfji M , Al-Mallah M H 
Published: October 2022

Abstract

Introduction
Cardiovascular magnetic resonance (CMR) is the gold standard for determination of left ventricular volumes (LVV), including end diastolic volume (LVEDV) and end systolic volume (LVESV). With incorporation of ECG gating, the quantitative analysis of ventricular function can be achieved in Positron Emission Tomography (PET) studies. However, the accuracy of LVV quantification using PET has not been shown in previous studies.

Purpose
We aimed to assess the inter-study variability of LVEDV and LVESV between CMR and PET.

Methods
Consecutive patients who underwent both PET and CMR within 1 year were identified from prospective institutional registries. LVV on PET was compared against CMR as a reference standard using several statistical measures of agreement.

Results
The study population consisted of 233 patients (mean age 60.8±13.6 years, 64.8% male). The median (IQR) duration between CMR and PET was 41 (11–96) days. The median (IQR) LVEDV values were 144 (107–196) on PET and 178 (138–236) on CMR (mean difference 36.09±2.36, p&lt;0.001). As shown in Figure 1, there was a good correlation (Spearman rho=0.88, p&lt;0.001; Intraclass Correlation Coefficient 0.88, 95% CI 0.84–0.90, P&lt;0.001) with moderate limits of agreement (95% limits of agreement −108.17–35.99). A total of 0% and 3.9% of the cohort had LVEDV &gt;150 ml/m2 by CMR and PET respectively. Overall results were similar with LVESV values and similar in subgroup analysis of patients with a low EF (&lt;40%), a coronary artery disease scar, LV hypertrophy and of patients with defibrillators.

Conclusion
Our analysis has shown a good correlation of LVV between PET and CMR, indicating PET derived LVV can accurately be used in the diagnosis and management of patients.

Funding Acknowledgement
Type of funding sources: None.
Early stage atherosclerosis regression using combined sonotherapy and mitoxantrone-mediated sonoporation therapy
Authors: Mehrad H 
Published: October 2022

Abstract

Background
In atherosclerosis, local inflammation and associated macrophage activity can lead to foam cell-rich plaque formation, which results to plaque rupture and thrombosis, making inflammation an important therapeutic target in cardiovascular disease. The early stage of atherosclerosis is characterized by the accumulation of low-density lipoprotein (LDL) droplets, leading to the creation of foam cells. Foam cells are generated by uncontrolled uptake of modified LDL, especially oxidized LDL (oxLDL).

Purpose
In this study, we developed an experimental focused ultrasound-based combined sonotherapy and sonoporation therapy system, and investigated its effectiveness on macrophage-derived foam cells reduction, wherein a high resolution diagnostic ultrasound is adjuncted with combination therapy system, with a goal of increased safety.

Methods
Briefly, New Zealand white rabbits underwent primary balloon dilatation injury at the right common carotid artery followed by a 1.5% cholesterol-rich diet injury for three weeks. Histopathology results showed the early stage atherosclerosis formation in all of the rabbits' arteries. Then common carotid arteries of the treatment group at the lesion region, treated using extracorporeal low-level pulsed-focused ultrasound (P=11 W, F=1.1 MHz, DF=50%) sonotherapy accompanied by mitoxantrone-loaded PESDA (Perfluorocarbon-Exposed Sonicated Dextrose Albumin) microbubbles (100 ml/kg, 2–5x105 bubbles/ml)-mediated sonoporation therapy. Foam cells density were evaluated in the treatment group compared with the other groups using B-mode ultrasonography and histopathology.

Results
Results from B-mode ultrasonography and histopathology showed a significant reduction in the mean value for immune cells and foam cells density within the early stage atherosclerotic lesion in the treatment group compared with the other groups (p&lt;0.05).

Conclusion
Enhanced sonoporation effect of ultrasound, induced by collapsed PESDA microbubbles accompanied by anti-inflammatory effect of sonotherapy and antineoplastic and cytotoxic effect of mitoxantrone can cause to reduce the immune cells and foam cells density within the lesion. These finding provide the basis for developing low-level pulsed-focused ultrasound-based combined sonotherapy and sonoporation therapy for a successful clinical application in the treatment of the early stage atherosclerosis.

Funding Acknowledgement
Type of funding sources: Public Institution(s). Main funding source(s): Mehrad Research Lab
Coronary microvascular disease assessed by 82Rb-PET-CT is an independent prognostic marker of all-cause mortality
Authors: Hojstrup S , Hansen K , Talleruphuus U , Marner L , Galatius S , Rauf M , Bjerking L , Jakobsen L , Christiansen E , Bouchelouche K , Prescott E 
Published: October 2022

Abstract

Background
Coronary microvascular disease (CMD) is a major contributor to e.g. heart failure and angina pectoris, as well as being associated with an increased risk of adverse events. CMD is diagnosed by reduced myocardial blood flow reserve (MBFR), preferably by Positron emission tomography myocardial perfusion CT (PET-CT).

Purpose
We aim to determine whether reduced MBFR is associated with an increased hazard of all-cause mortality independently of the extent of perfusion defects in patients suspected of obstructive coronary artery disease.

Method
We conducted a multicenter study of all patients referred for 82Rubidium PET-CT imaging between January 2018 and August 2020. Rest and stress examinations were performed using standard imaging protocols. Percentage of perfusion defects were calculated based on summed rest- and difference score. CMD was defined as MBFR ≤2. Patients were followed for all-cause mortality through national registries with no loss to follow-up.

Results
Among the 7156 patients studied, 61.8% were men, median age was 69 [61–76 IQR] years, 14.1% had LVEF ≤40%, 58.4% had a previous diagnosis of ischemic heart disease (IHD), 20.1% had atrial fibrillation and 38.9% had MBFR ≤2.
A total of 571 (7.8) deaths were observed, more frequently in MBFR ≤2 compared to MBFR &gt;2 (4.2% vs 13.2%, p&lt;0.001). MBFR was significantly associated with reversible hypoperfusion (r2=−0.33, p&lt;0.0001). In Kaplan-Meier estimation MBFR ≤2 was significantly associated to all-cause mortality in the overall population as well as in clinically relevant subgroups defined by the extent of reversible and/or irreversible perfusion defects (p&lt;0.05 for all, fig. 1). In multivariate Cox-analysis adjusting for age, sex, Charlson's Co-morbidity index, eGFR, LVEF and LVEF-reserve and stratifying by diabetes, MBFR ≤2 remained a robust predictor of all-cause mortality with a HR 1.73, 95% CI: 1.62–2.19, p&lt;0.0001 (fig. 2). No interaction was found between MBFR and reversible hypoperfusion. In subgroup analysis including only patients with no reversible perfusion defects (n=3095), MBFR ≤2 was still strongly associated with a HR of 2.00, 95% CI: 1.29–3.11, p&lt;0.001 for all-cause mortality.

Conclusion
MBFR ≤2 is a robust predictor of all-cause mortality independently of the extent of reversible- and/or irreversible perfusion defects. Information of MBFR should be incorporated in the clinical risk stratification of patients being investigated for ischemia.

Funding Acknowledgement
Type of funding sources: Public hospital(s). Main funding source(s): Research Committee, Bispebjerg & Frederiksberg University Hospital, scientific scholarship
SGLT-2 inhibition by empagliflozin exerts neutral effects on experimental arterial thrombosis in a murine model of low-grade inflammation
Authors: Liberale L , Kraler S , Puspitasari Y , Bonetti N , Akhmedov A , Ministrini S , Montecucco F , Marx N , Lehrke M , Hartmann N U K , Beer J H , Paneni F , Luescher T F , Camici G G 
Published: October 2022

Abstract

Introduction
Low-grade inflammation couples dysmetabolic states to insulin resistance and atherosclerotic cardiovascular disease (ASCVD). Selective sodium-glucose co-transporter 2 (SGLT-2) inhibition by empagliflozin improves clinical outcomes in patients with ASCVD independently of glucose-lowering. Yet, its mechanism of action remains largely undetermined.

Purpose
We aimed to test whether empagliflozin affects arterial thrombus formation in baseline conditions or low-grade inflammation, a systemic milieu shared among patients with ASCVD.

Methods
Sixteen-week-old C57BL/6 mice were randomly assigned to acute administration of empagliflozin (25 mg/kg BW) or vehicle, of which a subgroup was pre-treated biweekly over 4 weeks with super-low-dose lipopolysaccharide (LPS; 5 ng/kg BW), before carotid thrombosis was induced by photochemical injury. The translational value of these findings was investigated in primary human aortic endothelial cells (HAECs) and plasma samples of patients randomized to empagliflozin therapy.

Results
The between-group difference in doppler-flow probe detected time-to-occlusion (TTO) remained within the predefined equivalence margin (Δ=|10.50|), irrespective of low-grade inflammation (95% confidence interval [CI], −9.82 to 8.85 and −9.20 to 9.69), while glucose dropped by 1.64 and 4.84 mmol/l, respectively (Fig. 1). Ex vivo platelet aggregometry suggests similar platelet activation status, corroborated by unchanged circulating platelet-factor 4 (PF4) plasma levels. In concert, carotid PAI-1 expression and TF activity remained unaltered upon SGLT-2 inhibition, and no difference in plasma D-dimer levels was detected, suggesting comparable coagulation cascade activation and fibrinolytic activity (Fig. 1). In HAECs pre-treated with LPS, empagliflozin neither changed TF activity nor PAI-1 expression (Fig. 2). Accordingly, among patients with established ASCVD or at high cardiovascular (CV) risk randomized to 10 mg empagliflozin daily signatures of thrombotic (i.e., TF) and fibrinolytic activity (i.e., PAI-1) remained unchanged, while plasma glucose declined significantly during 3 months of follow-up (Fig. 2).

Conclusion
SGLT-2 inhibition by empagliflozin does not impact experimental arterial thrombus formation, neither under baseline conditions nor during sustained low-grade inflammation, and has no impact on proxies of thrombotic/fibrinolytic activity in patients with ASCVD. The beneficial pleiotropic effects of empagliflozin are likely independent of pathways mediating thrombosis.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Swiss National Science FoundationSwiss Heart FoundationFoundation for Cardiovascular Research–Zurich Heart House
Thrombosis-related miR-16-5p predicts the disease severity in patients hospitalised for COVID-19
Authors: Keshwani D , Eyileten C , Wicik Z , Nowak A , Jakubik D , Simoes S N , Martins-Jr D C , Shahzadi A , Jarosz-Popek J , Wolska M , Siller-Matula J , Postula M 
Published: October 2022

Abstract

Introduction
SARS-CoV-2 tropism for the ACE2 receptor, along with the multifaceted inflammatory reaction, is likely to drive the generalized hypercoagulable state seen in patients with COVID-19.

Methodology
Using the original bioinformatic workflow and network medicine approaches we reanalyzed four coronavirus-related expression datasets and performed co-expression analysis focused on thrombosis and ACE2 related genes. We identified microRNAs (miRNAs) which play role in ACE2-related thrombosis in coronavirus infection and further, we validated the expressions of those miRNAs in 79 hospitalized COVID-19 patients and 32 healthy volunteers by PCR and monitored miRNAs patterns during the acute phase of COVID-19, as well as the prognostic potential of these miRNAs as biomarkers.

Results
We identified EGFR, HSP90AA1, APP, TP53, PTEN, UBC, FN1, ELAVL1 and CALM1 as regulatory genes which could play a pivotal role in COVID-19 related thrombosis. We also found miR-16-5p, miR-27a-3p, Let-7b-5p and miR-155-5p as regulators in coagulation and thrombosis process. We observed in separate cohort of COVID-19 patients and healthy controls that (i) expression of miR-16-5p, miR-27a-3p and miR-155-5p increased during observation, compared to the baseline measurement; (ii) a low baseline miR-16-5p expression presents predictive utility in assessment of the hospital length of stay or death in follow-up as a composite endpoint (AUC: 0.810, 95% CI, 0.71–0.91, p&lt;0.0001); (iii) low baseline expression of miR-16-5p and diabetes mellitus are independent predictors of increased length of stay or death according to a multivariate analysis (OR: 9.417; 95% CI, 2.647–33.506; p=0.0005 and OR: 6.257; 95% CI, 1.049–37.316; p=0.044, respectively).

Conclusion
This study enabled us to better characterize changes in gene expression and signaling pathways related to COVID-19 thrombosis. In this study we identified, characterized and validated miRNAs which could serve as novel, thrombosis-related biomarkers of the COVID-19, can be used for early stratification of patients and prediction of severity of infection development in an individual.

Funding Acknowledgement
Type of funding sources: Public Institution(s). Main funding source(s): Medical University of Warsaw
Citrullination of histoneH3 in neutrophil via CXCL1 enhances neutrophil adhesion to femoral artery of LDLR−/− mice fed HFD
Authors: Osaka M , Yoshida M 
Published: October 2022

Abstract

Background
Vascular inflammation plays an important role in the development of atherosclerosis. Previously we have shown that a high-fat diet (HFD) increased neutrophil adhesion to the vascular intima in wild-type (wt) mice (Osaka M. Sci Rep. 2016). However, the involvement of neutrophils in atherosclerosis-related vascular inflammation is not well known.

Purpose
This study examined that neutrophil extracellular trap (NETs) or the hypercitrullination of histone H3 in neutrophils enhances neutrophil adhesion to atheroprone-arteries in LDL receptor null (LDLR−/−) mice.

Methods
We observed leukocyte adhesion in the femoral artery of LDLR−/− mice fed normal chow (NC) or HFD, and determined leukocyte subtype that adhered on vascular endothelium under neutrophil or monocyte depletion using intravital microscopy. Importantly, neutrophil adhesion was examined under the administration of TDFA which inhibits NETs and citrullination of histone H3, in LDLR−/− mice fed HFD. Furthermore, immunohistochemistry for citrullinated histone H3 in peripheral neutrophils of mice was examined. Comprehensive cytokine/chemokine analysis for a plasma of mice was performed to determine the factors citrullinating histone H3 in LDLR−/− mice. Moreover, these mice were treated with a novel specific PPARα agonist, to reduce the elevation of plasma triglyceride levels.

Results
Leukocyte adhesion in LDLR−/− mice fed HFD significantly increased compared to NC. More interestingly, it significantly enhanced compared to wt mice fed HFD. Furthermore, neutrophil depletion rather than monocyte depletion diminished leukocyte adhesion, suggesting that the leukocyte subtype that adhered in LDLR−/− mice fed HFD was neutrophil. Neutrophil adhesion in these mice significantly was reduced by the administration of TDFA, suggesting a pivotal role for histone H3 citrullination in neutrophil adhesion. Moreover, citrullination of histone H3 in neutrophils from LDLR−/− mice fed HFD but not from those without HFD was significantly enhanced. In addition, comprehensive cytokine/chemokine analysis revealed an increase of CXCL1 in plasma of LDLR−/− mice fed HFD. CXCL1 enhanced neutrophil adhesion to HUVECs, and the adhesion significantly decreased by the treatment of TDFA to neutrophil in vitro non-static adhesion assay. These results showed that CXCL1 enhanced neutrophil adhesion in LDLR−/− mice fed HFD through citrullination. Furthermore, when these mice were treated with PPARα agonist, observed histone citrullination, as well as neutrophil adhesion, was significantly reduced.

Conclusion
These results suggest that HFD induced histone citrullination in neutrophils in LDLR−/− mice and PPARα agonist plays a role during hypertriglyceridemia-mediated vascular inflammation in atherosclerosis.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Grant-in-Aid for Scientific Research(C)