bluuebunny/crossref_metadata_2025_split huggingface dataset:
Beta3-adrenergic stimulation restores endothelial mitochondrial dynamics and prevents pulmonary arterial hypertension
Authors: Oliver E , Rocha S.F , Spaczynska M , Lalama D.V , Gomez M , Fuster V , Ibanez B 
Published: November 2020

Abstract

Background
Endothelial dysfunction is one of the most important hallmarks of pulmonary arterial hypertension (PAH). This leads to anomalous production of vasoactive mediators that are responsible for a higher vascular tone and a subsequent increase in pulmonary artery pressure (PAP), and to an increased vascular permeability that favors perivascular inflammation and remodeling, thus worsening the disease. Therefore, preservation of the endothelial barrier could become a relevant therapeutic strategy.

Purpose
In previous studies, others and we have suggested the pharmacological activation of the β3-adrenergic receptor (AR) as a potential therapeutic strategy for pulmonary hypertension (PH) due to left heart disease. However, its potential use in other forms of PH remain unclear. The aim of the present study was to elucidate whether the β3-AR agonist mirabegron could preserve pulmonary endothelium function and be a potential new therapy in PAH.

Methods
For this purpose, we have evaluated the effect of mirabegron (2 and 10 mg/kg·day) in different animal models, including the monocrotaline and the hypoxia-induced PAH models in rats and mice, respectively. Additionally, we have used a transgenic mouse model with endothelial overexpression of human β3-AR in a knockout background, and performed in vitro experiments with human pulmonary artery endothelial cells (HPAECs) for mechanistic experiments.

Results
Our results show a dose dependent effect of mirabegron in reducing mean PAP and Right Ventricular Systolic Pressure in both mice and rats. In addition, the use of transgenic mice has allowed us to determine that pulmonary endothelial cells are key mediators of the beneficial role of β3-AR pathway in ameliorating PAH. Mechanistically, we have shown in vitro that activation of β3-AR with mirabegron protects HPAECs from hypoxia-induced ROS production and mitochondrial fragmentation by restoring mitochondrial fission/fusion dynamics.

Conclusions
This protective effect of mirabegron would lead to endothelium integrity and preserved pulmonary endothelial function, which are necessary for a correct vasodilation, avoiding increased permeability and remodeling. Altogether, the current study demonstrates a beneficial effect of the β3-AR agonist mirabegron that could open new therapeutic avenues in PAH.

Funding Acknowledgement
Type of funding source: Public grant(s) – National budget only. Main funding source(s): Programa de Atracciόn de Talento, Comunidad de Madrid
Pulmonary arterial hypertension modeling with synthetic stable thromboxane A2 analogue (U46619) in porcine: dose adjustment and acute hemodynamic reactions
Authors: Goncharova N , Andreeva E , Vakhrushev A , Condori Leandro H.I , Murashova L , Voronin S , Korobchenko L , Mitrofanova L , Moiseeva O , Mikhaylov E 
Published: November 2020

Abstract

Introduction
Survival in PAH is strongly associated with adaptation of the RV to pressure overload.

Purpose
To investigate acute hemodynamic reactions during infusion of the stable thromboxane A2 (TXA2) analogue U46619 in a porcine model.

Materials and methods
The study comprised 9 male Landrace pigs (32.7±3.8 kg) under GA. After heparinization, PAH was induced by continuous and stepwise increasing infusion of U46619 (10 mg/ml; Tocris, USA) according to a pre-specified protocol. The target mPAP was 40 mmHg. Hemodynamics was assessed, cardiac output (CO) was calculated using the Fick equation before and during U46619 infusion.

Results
U46619 infusion at a previously published rate 0.1, 0.2 μg kg–1 min–1 in pig 1 resulted in rapid hemodynamic deterioration. Two-time reduced U46619 dose infusione resulted in a target mPAP but severe mBP drop in 1, 3, 4 pigs and required cathecholamine support. The U46619 dosage was reduced up to ¼ (0.025, 0.05, 0.075, 0.1, 0.12, 0.15, 0.175 μg kg–1 min–1). In animals on 1/4 dosage U46619 infusion, heart rate (HR), mBP, mPAP, RAP and PVR reliably increased and BP was stable (Table 1). There was a positive correlation between the mPAP and heart rate (HR) on U46619 infusion (r=0.66; t=2.38; p=0.048) and between CO and mBP (r=0.66; t=2.36; p=0.04). Low CO was associated with high PVR (r=−0.98; t=−14.3; p&lt;0,001). Positive correlation between PVR and SVR was revealed (r=0.96; p=0.00002). In 3 pigs with severe mBP drop and PAH right ventricle subendocardial hemorrhage was revealed on autopsy study.

Conclusions
High dose U46619 infusion was associated with acute RV decompensation due to pressure overload accompanied with systemic BP drop and low CO. Lower dosages of U46619 infusion were characterized by stable target mean PAP. HR acceleration and mean systemic BP are of compensation mechanisms for cardiac output maintenance in PAH.

Funding Acknowledgement
Type of funding source: Public grant(s) – National budget only. Main funding source(s): RFBR grant 18-315-20050
Shear stress induced activation of von Willebrand factor may predispose to gastrointestinal bleeding in syndrome of Heyde
Authors: Avtaeva Y.N , Melnikov I.S , Komlev A.E , Imaev T.E , Okhota S.D , Kozlov S.G , Gabbasov Z.A 
Published: November 2020

Abstract

Introduction
Von Willebrand factor (vWF) changes conformation from globular to fibrillar in the range of shear rates above 5000 s-1. High shear rates, observed in severe aortic stenosis, create conditions for activation of vWF, which opens up access for platelets and coagulation factors to the previously hidden domains of the molecule. At the same time, vWF undergoes increased degradation by metalloproteinase ADAMTS13. Proteolytic cleavage of vWF leads to deficiency of hemostatically active high molecular weight multimers (HMWM) of vWF, while its mass concentration remains unaltered. This results in the development of acquired von Willebrand disease type 2A (vWD) and concomitant gastrointestinal bleeding. The combination of acquired vWD 2A and gastrointestinal bleeding, developed due to severe aortic stenosis, is called Heyde's syndrome. The correlation of shear stress activation of vWF and gastrointestinal bleeding in patients with Heyde's syndrome remains poorly studied.
The aim of the study was to measure vWF-mediated platelet adhesion to fibrinogen-coated surfaces under shear rates higher than 5000 s-1 in whole blood samples of healthy volunteers and patients with Heyde's syndrome.

Methods
A microfluidic system simulating blood flow in vessels was used to assess platelet adhesion. Platelet adhesion was measured by an increase in the intensity of laser radiation scattered from a fibrinogen-coated surface during a 15 minutes circulation of whole blood samples through a flow chamber under shear rates higher than 5000 s-1. Platelets in whole blood samples were activated with 5 μM ADP prior to measurement. The study included 5 patients with Heyde's syndrome 55–80 years old. The control group included 6 healthy volunteers 25–55 years old. vWF-mediated platelet adhesion was detected by blocking platelet-vWF binding with anti-GPIb monoclonal antibody (mAb). Fibrinogen-mediated platelet adhesion was detected by blocking platelet GPIIb/IIIa receptors with mAb.

Result
The inhibition of GPIb vWF-receptor reduced platelet adhesion by 7.6±3.5% (p&lt;0.05) in patients with Heyde's syndrome, and by 16.5±3.3% (p&lt;0.05) in healthy volunteers. The inhibition of GPIIb/IIIa fibrinogen receptor reduced platelet adhesion by 96±7% (p&lt;0.05) in patients with Heyde's syndrome, and by 80.2±6.6% (p&lt;0.05) in healthy volunteers.

Conclusion
Significantly reduced contribution of vWF to platelet adhesion under shear rates higher than 5000 s-1 may indicate a decrease in hemostatically active HMWM of vWF. Shear stress activation of vWF in the range of high shear rates and its subsequent inactivation by ADAMTS13 may lead to functional vWF deficiency and the development of gastrointestinal bleeding in Heyde's syndrome.

Funding Acknowledgement
Type of funding source: Foundation. Main funding source(s): The Russian Science Foundation
Shaping the future of metabolic syndrome: genetics, prognosis and individual tailoring
Authors: Sousa J.A , Mendonca M.I , Pereira A , Monteiro J.P , Temtem A.M , Santos M , Mendonca F.V , Sousa A.C , Rodrigues M , Henriques E , Ornelas I , Freitas A.I , Freitas A.D , Reis P 
Published: November 2020

Abstract

Background
Metabolic syndrome (MetS), characterized by a cluster of cardiovascular risk factors, is considered to be the major health hazard of modern world and a 21st century epidemic. Recent GWAS have identified several susceptibility regions involved in lipid metabolism and oxidation, also associated with MetS. Genetic risk score (GRS) is an emerging method that attempts to establish correlation between SNPs and clinical phenotypes.

Aim
Evaluate the value of a GRS encompassing SNPs involved in lipidic metabolism and oxidation pathways, in predicting CAD outcome (MACEs and long-term cardiovascular Mortality) in a coronary population with MetS.

Methods
1101 coronary patients and MetS, were selected from the GENEMACOR study. Genotyping was performed by TaqMan allelic discrimination assay. A Multiplicative score (mGRS) was constructed according to the multiplicative model with variants belonging to the lipid and oxidative axes (PSRC1, PCSK9, KIF6, ZNF259, LPA, APO E, PON192, PON108, PON55, MTHFR677, MTHFR1298, MTHFD1L). This GRS was categorized using the mean (higher vs lower than mean). Cumulate Mortality Hazards Model (Cox regression) adjusted for age, gender, smoking, hypertension, dyslipidaemia, diabetes, hsCRP, eGFR, Ejection fraction (EF), and multivessel disease) was used to find independent predictors of cardiovascular outcome. We performed Kaplan-Meier Survival curves for both groups (higher vs lower than mean GRS) and log-rank test to compare survival distributions in both groups.

Results
The following variables have emerged independently associated with time to MACE occurrence: mGRS (HR=1.31 95% CI (1.07; 1.59); p=0.008), male gender, EF and multivessel disease. Concerning cardiovascular mortality, mGRS also remained an independent predictor (HR=1.44 (1.04–1.99); p=0.028) alongside age, smoking, diabetes and EF. The Log-Rank test showed significant differences between the two curves related to MACE occurrence and cardiovascular mortality (p=0.001 and 0.002, respectively). The Kaplan-Meier survival showed that as mGRS increases, patient survival decreases.

Conclusion
In patients with MetS, a GRS comprising variants in lipidic and oxidative pathways, proved to be a useful stratification tool, identifying patients likely to have a worst prognosis over time. Our data further underlines the additive potential and clinical utility of genetic information in shaping secondary prevention.
Figure 1

Funding Acknowledgement
Type of funding source: None
An increase in activated NKT-CD8+ cells is associated with the presence of hypoechoic carotid plaques
Authors: Genkel V , Baturina I , Emelyanov I , Shaposhnik I , Dolgushin I 
Published: November 2020

Abstract

Background
The role of NK-cells and NKT-cells and their subtypes at various stages of the development of atherosclerosis is poorly understood. It was previously shown that CD3+CD56+CD8+ cells can secrete pro-inflammatory cytokines and an increase in their number has been established in patients with active Behçet's Disease and Multiple Sclerosis.

Purpose
To study the relationship between the number of activated CD3+CD56+CD8+ cells and the echogenicity of plaques in patients with carotid atherosclerosis.

Methods
The study included 40 patients, 21 (52.5%) men and 19 (47.5%) women who underwent carotid ultrasound and immunological studies. The median age was 53.5 (46.7; 57.0) years. All patients underwent duplex ultrasound scanning of the carotid arteries. If an atherosclerotic plaque was detected, its image was saved and exported to Adobe Photoshop CS6® followed by analysis of the gray-scale median (GSM) of the plaque. Hypoechoic was considered a carotid plaque whose GSM values were less than 50. The number of CD3+CD56+CD8+CD11b+ cells was evaluated by flow cytometry using a flow cytometer.

Results
Among the examined patients, carotid plaques were detected in 31 (77.5%) participants. The average values of GSM visualized plaques were 58.0 (46.0; 71.5). Atherosclerotic plaques with GSM &lt;50 were detected in 11 (35.5%) patients.
In patients with hypoechoic carotid plaques, the number of CD3+CD56+CD8+CD11b+ cells was statistically significantly higher – 352 (55.0; 505) cells/uL versus 66.0 (15.5; 138) cells/uL in patients with plaques whose GSM values were more than 50 (p=0.020). Based on the results of correlation analysis it was established inverse relationship between number of CD3+CD56+CD8+CD11b+ cells and GSM of plaques (r=−0.427; p=0.016). According to the results of the ROC-analysis, an increase in the number of activated NKT-CD8+ cells over 46.5 cells/uL was a predictor of the presence of hypoechogenic carotid plaques with a sensitivity of 81.8% and a specificity of 45.0% (see Figure 1).

Conclusion
In patients with hypoechoic carotid plaques, the number of CD3+CD56+CD8+CD11b+ cells was significantly higher in comparison with other patients. An increase in the number of activated NKT-CD8+ cells over 46.5 cells/uL was a predictor of the presence of hypoechoic carotid plaques with sensitivity of 81.8% and specificity of 45.0%
Figure 1

Funding Acknowledgement
Type of funding source: None
Empagliflozin impairs smooth muscle cell proliferation, accelerates endothelial regeneration, and prevents neointima formation by altered expression of the vasoactive peptide apelin
Authors: Dutzmann J , Bode L.M , Korte L , Kalies K , Koch S , Tongers J , Bauersachs J , Sedding D.G 
Published: November 2020

Abstract

Background
Empagliflozin, an inhibitor of the sodium glucose co-transporter 2 (SGLT2), developed as an anti-diabetic agent exerts additional beneficial effects on heart failure outcomes in patients with type 2 diabetes mellitus at high cardiovascular risk. However, the effect of empagliflozin on vascular cell function and vascular remodeling processes remain largely elusive.

Methods/Results
Immunocytochemistry and immunoblotting revealed SGLT2 to be expressed in human diabetic and non-diabetic smooth muscle (SMC) and endothelial cells (EC) as well as in murine femoral arteries. In vitro, empagliflozin significantly reduced serum-induced proliferation and migration of human diabetic and non-diabetic SMCs in a dose-dependent manner without any toxic or apoptotic effects. In contrast, empagliflozin significantly increased the cell count and migrational capacity of human diabetic ECs, but not of human non-diabetic ECs. In vivo, therapeutic application of empagliflozin (225 mg/kg medicated diet) resulted in a significantly reduced number of Ki-67+ proliferating neointimal cells in response to femoral artery wire-injury in C57BL/6J mice. Empagliflozin furthermore prevented subsequent neointima formation (luminal stenosis 91.2% vs. 80.6% at 21 days; P&lt;0.05). Comparable effects of empagliflozin were observed in a streptozocin-induced diabetic model of apolipoprotein E−/− mice. Conclusive to the in vitro-results, re-endothelialization was not significantly affected in C57BL/6 mice (non-reendothelialized distance 2.57 mm vs. 2.3 mm; P=0.07), but even significantly improved in diabetic mice after treatment with empagliflozin (3.1 mm vs. 2.58 mm; P&lt;0.001) assessed by Evan's Blue staining 3 days after electric denudation of the murine carotid artery.
Microarray analysis of human SMCs identified the vasoactive peptide apelin to be decisively regulated in response to empagliflozin treatment. Further pathway analysis exhibited apelin to prevent SMC proliferation by de-phosphorylation of Akt and to augment EC proliferation by phosphorylation of p38 MAPK.

Conclusion
These data document the functional impact of empagliflozin on vascular SMCs and ECs for the first time. Empagliflozin significantly reduces serum-induced proliferation and migration of SMCs in vitro and prevented neointima formation in vivo, while augmenting EC proliferation in vitro and re-endothelialization in vivo after vascular injury. Thus, empagliflozin holds promise to exert favorable effects on vascular healing, and to prevent neointima formation following vascular injury in diabetic and non-diabetic patients.

Funding Acknowledgement
Type of funding source: Public Institution(s). Main funding source(s): Hannover Medical School, Martin-Luther-University Halle-Wittenberg
Empagliflozin and health-related quality of life outcomes in patients with heart failure with reduced ejection fraction: the EMPEROR-Reduced trial
Authors: Butler Javed , Anker Stefan D , Filippatos Gerasimos , Khan Muhammad Shahzeb , Ferreira João Pedro , Pocock Stuart J , Giannetti Nadia , Januzzi James L , Piña Ileana L , Lam Carolyn S P , Ponikowski Piotr , Sattar Naveed , Verma Subodh , Brueckmann Martina , Jamal Waheed , Vedin Ola , Peil Barbara , Zeller Cordula , Zannad Faiez , Packer Milton ,   the EMPEROR-Reduced Trial Committees and Investigators
Published: November 2020

Abstract

Aims
In this secondary analysis of the EMPEROR-Reduced trial, we sought to evaluate whether the benefits of empagliflozin varied by baseline health status and how empagliflozin impacted patient-reported outcomes in patients with heart failure with reduced ejection fraction.

Methods and results
Health status was assessed by the Kansas City Cardiomyopathy Questionnaires-clinical summary score (KCCQ-CSS). The influence of baseline KCCQ-CSS (analyzed by tertiles) on the effect of empagliflozin on major outcomes was examined using Cox proportional hazards models. Responder analyses were performed to assess the odds of improvement and deterioration in KCCQ scores related to treatment with empagliflozin. Empagliflozin reduced the primary outcome of cardiovascular death or heart failure hospitalization regardless of baseline KCCQ-CSS tertiles [hazard ratio (HR) 0.83 (0.68–1.02), HR 0.74 (0.58–0.94), and HR 0.61 (0.46–0.82) for &lt;62.5, 62.6–85.4, and ≥85.4 score tertiles, respectively; P-trend = 0.10]. Empagliflozin improved KCCQ-CSS, total symptom score, and overall summary score at 3, 8, and 12 months. More patients on empagliflozin had ≥5-point [odds ratio (OR) 1.20 (1.05–1.37)], 10-point [OR 1.26 (1.10–1.44)], and 15-point [OR 1.29 (1.12–1.48)] improvement and fewer had ≥5-point [OR 0.75 (0.64–0.87)] deterioration in KCCQ-CSS at 3 months. These benefits were sustained at 8 and 12 months and were similar for other KCCQ domains.

Conclusion
Empagliflozin improved cardiovascular death or heart failure hospitalization risk across the range of baseline health status. Empagliflozin improved health status across various domains, and this benefit was sustained during long-term follow-up.

Clinical trial registration
URL: https://www.clinicaltrials.gov. Unique identifier: NCT03057977.
Post-natal developmental changes in the composition of the rat neocortical N-glycome
Authors: Klarić Thomas S , Salopek Matija , Micek Vedran , Gornik Kljaić Olga , Lauc Gordan 
Published: November 2020

Abstract
Asparagine-linked glycosylation (N-glycosylation) plays a key role in many neurodevelopmental processes, including neural cell adhesion, neurite outgrowth and axon targeting. However, little is known about the dynamics of N-glycosylation during brain development and, in particular, how the N-glycome of the developing neocortex differs from that of the adult. The aim of this study, therefore, was to perform a thorough characterization of N-glycosylation in both the adult and neonatal rat neocortex in order to gain insights into the types of changes occurring in the N-glycome during neurodevelopment. To this end, we used hydrophilic interaction ultraperformance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight mass spectrometry to compare the adult neocortical N-glycome with that of 24- and 48-h neonates. We report that the abundance of complex N-glycans is significantly lower in adults compared with neonates. Furthermore, the proportion of charged complex N-glycans is also greatly reduced. This decrease in the abundance of complex N-glycans is offset by a corresponding increase in the proportion of truncated and, to a lesser extent, hybrid N-glycans. Lastly, we report that although the proportion of oligomannose N-glycans remains constant at around 24%, the distribution of high-mannose subtypes shifts from predominantly large subtypes in neonates to smaller subtypes in the adult. In summary, our findings indicate that N-glycan synthesis in the rat neocortex is fundamentally different in neonates compared with adults with a general shift occurring from large, sialylated N-glycans towards smaller, neutral structures as neonates develop into adults, coupled with a parallel shift towards smaller oligomannose structures.
Transient establishment of imprinted DNA methylation of transgenic human IC1 sequence in mouse during the preimplantation period
Authors: Hirakawa Katsuhiko , Matsuzaki Hitomi , Tanimoto Keiji 
Published: November 2020

Abstract
Monoallelic gene expression at the Igf2/H19 locus is controlled by paternal allele-specific DNA methylation of the imprinting control region (H19 ICR) that is established during spermatogenesis. We demonstrated that the H19 ICR fragment in transgenic mice acquires allele-specific methylation only after fertilization, which is essential for maintaining its allelic methylation during early embryogenesis. We identified a DNA element required for establishing postfertilization methylation within a 118 bp (m118) region. A previously generated knock-in mouse whose endogenous H19 ICR was substituted with the human H19 ICR (hIC1; 4.8 kb) sequence revealed that the hIC1 sequence was partially methylated in sperm, although this methylation was lost by the blastocyst stage, which we assume is due to a lack of an m118-equivalent sequence in the hIC1 transgene.
To identify a cis sequence involved in postfertilization methylation within the hIC1 region, we generated three transgenic mouse lines (TgM): one carrying an 8.8 kb hIC1 sequence joined to m118 (hIC1+m118), one with the 8.8 kb hIC1 and one with the 5.8 kb hIC1 sequence joined to m118 (hIC1–3′+m118). We found that the hIC1–3′ region was resistant to de novo DNA methylation throughout development. In contrast, the 5′ portion of the hIC1 (hIC1–5′) in both hIC1+m118 and hIC1 TgM were preferentially methylated on the paternal allele only during preimplantation. As DNA methylation levels were higher in hIC1+m118, the m118 sequence could also induce imprinted methylation of the human sequence. Most importantly, the hIC1–5′ sequence appears to possess an activity equivalent to that of m118.