bluuebunny/crossref_metadata_2025_split huggingface dataset:
Metabolic regulation of endothelial reactive oxygen species production in obesity
Authors: Padgett C , Speese A , Rosewater C , Corley Z , Mintz J , Fulton D , Stepp D 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): National Institutes of Health (NIH)

Introduction
The prevalence of obesity continues to rise to unprecedented levels, and with it, a corresponding increase in the incidence of cardiovascular disease (CVD). While obesity is indeed accepted as one of the most prominent risk factors for CVD, the precise molecular mechanisms by which aberrant metabolism is linked to cardiovascular function remain incompletely understood. One prevailing hypothesis is that overproduction of reactive oxygen species (ROS) from increased metabolism significantly contributes to endothelial dysfunction, which precedes many cardiovascular events such as atherosclerosis and stroke.

Purpose
In this study, we hypothesized that a receptor for advanced glycation end products, Galectin-3 (GAL3), acts as a metabolic sensor and regulates endothelial ROS production in obesity.

Methods
Obese db/db mice were crossed with mice lacking GAL3, and endothelial gene expression, microvascular reactivity, and ROS production were assessed.

Results
We demonstrate that NADPH Oxidase I (NOX1), the predominant source of endothelial superoxide production, is down-regulated by GAL3 deletion, thereby rescuing endothelial function and ameliorating endothelial ROS production in obesity. Furthermore, we demonstrate that GAL3-mediated NOX1 over-expression is amenable to improvements in metabolic status, such as lowering blood glucose with metformin, improving glucose handling by augmenting muscle mass, or improving insulin signaling through deletion of Protein Tyrosine Phosphatase 1B (PTP1B).

Conclusion
Taken together, these data demonstrate that the overproduction of superoxide by endothelial NOX1 is regulated by the metabolic sensor GAL3 in obesity, leading to endothelial dysfunction and CVD. This pathway presents an attractive target for therapeutic intervention to break the link between aberrant metabolism in obesity and its corresponding vascular pathologies.
Assessing compartmentalisation of small non-coding RNAs by fractionation of lipoproteins and extracellular vesicles of human plasma
Authors: Teng I , Gutmann C , Takov K , Mayr M 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: None.

Background/Introduction
Circulating small non-coding RNAs (ncRNAs), such as microRNAs (miRNAs) and YRNAs, have been implicated in intercellular communication and as biomarkers for cardiometabolic diseases. The putative carriers of ncRNAs are lipoproteins, extracellular vesicles (EVs) and RNA-binding proteins. However, the lack of reliable separation methods for EVs and lipoproteins from human blood hampers the assignment of small ncRNA signatures to different carriers.

Purpose
To study which classes of ncRNAs are carried by EVs or lipoproteins in the circulation.

Methods & Results
In order to minimise the co-isolation of lipoproteins and EVs, size-exclusion chromatography (SEC) was established to improve resolution of nanoparticles from plasma. Mass spectrometry and immunoblotting for EV markers (syntenin-1 and CD9) and apolipoproteins (ApoA1 and ApoB for high- and low-density lipoproteins, respectively) were used to characterise the isolated preparations and identify co-isolating species. Quantitative real-time PCR of each carrier-rich fraction after SEC showed that miRNAs were mainly detected in the late-eluting fractions, enriched in ApoA1-containing lipoproteins. Only a small proportion of miRNAs was associated with fractions rich in albumin or in EVs. On the other hand, YRNAs were predominantly found in the albumin-rich and ApoB-containing fractions. To assess whether lipoproteins are key carriers for ncRNAs, HDL and ApoB immuno-depletion methods were employed. Both lipoprotein depletion methods failed to substantially deplete small ncRNAs from plasma.

Conclusion
We studied the distribution of circulating ncRNAs amongst different SEC fractions from human plasma. We have observed a minor role for lipoproteins as carriers of ncRNAs. Further investigation will focus on using SEC fractionation to assess EVs and RNA-binding proteins and their role as ncRNA carriers in plasma.
Elderberry extract inhibits tumour necrosis factor induced monocyte adhesion to endothelial cells via modulation of the NF-κB pathway
Authors: Festa J , Singh H , Hussain A , Da Boit M 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Other. Main funding source(s): De Montfort PhD funded scholarship scheme

Introduction
During the early stages of atherosclerosis, monocytes attach to endothelial cells (EC) before differentiating into macrophages causing the accumulation of foam cells and plaque growth. NF-κB and vascular cell adhesion molecule 1 (VCAM-1) are some of the main mediators in EC that are involved in the monocyte adhesion process. Elderberry extract (EE) are rich in anthocyanins a subclass polyphenol with vascular-protective properties.

Purpose
The main of aim of this study is to examine if EE can prevent TNF-α induced inflammation and monocyte adherence to EC.

Methods
Primary Human Umbilical Vein Endothelial Cells (HUVEC) were pre-treated for 1-h and stimulated with or without TNF-α 10 ng/ml for western blot. Cell lysates from the treated cells were then subjected to Western blotting and probed for total and phospho-NF-κB. For monocyte adherence cells were pre-treated for 24-h, then stimulated with/or without TNF-α 10 ng/ml for 24-h. Fluorescently labelled THP-1 cells were then added to the HUVECs for an additional 30 minutes, and samples were measured in a fluorescence plate reader. Cells were put through flow cytometry for measuring reactive oxygen species (ROS) using DCFHFDA assay or VCAM-1 levels using Anti-CD106.

Results
An increase in ROS production and NF-κB phosphorylation was found after stimulation TNF-α 10 ng/ml (p = 0.01). However, cells pre-incubated with EE (50 μg/ml) for 1-hour before TNF-α stimulation caused a reduction in ROS as well as inhibition of NF-κB phosphorylation (p &lt; 0.01). TNF-α 10 ng/ml increased the monocyte adherence to the HUVECs by a 2-fold although, EE prevented TNF-α monocyte adherence (mean value, 589.7 vs 408 p = 0.0033). This was associated with suppressed VCAM-1 expression found in the EE pre-treatment with TNF-α (p = 0.02).

Discussion
Our Preliminary data demonstrates that EE, can prevent monocytes binding onto EC potentially by inhibiting TNF-α induced NF-κB and VCAM-1 levels. Our findings postulate that NF-κB and VCAM-1 could be the direct link for targeting the prevention of monocyte adherence to EC, although this would need confirmation by blocking key cellular signalling pathways to confirm its role. This preliminary data suggests that EE and potentially other polyphenols could be a useful strategy for targeting the initial stages of atherosclerosis.
Serotype-specific microvascular changes in the liver due to dengue virus (DENV) non-structural protein-1 (NS1)
Authors: Duruanyanwu J , Campagnolo PC , Maringer KM 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public Institution(s). Main funding source(s): University of Surrey
Dengue virus is the most common arbovirus in humans worldwide, with a minor proportion developing severe dengue, which includes the cardiovascular complications of vascular leakage, haemorrhage, organ failure and shock, all of which could prove fatal. The mechanisms that lead to microvascular leakage remain incompletely understood. While DENV-2 NS1 has been shown by our group to disrupt endothelial-pericyte interactions leading to enhanced endothelial cell permeability in vitro, it is unclear whether this is true across all four serotypes of DENV and to what degree. It is also unclear whether DENV NS1 similarly affects all types of pericytes across the body. Our previous work was performed in saphenous venous pericytes, which we here extend to include hepatic stellate cells (HSCs), since the liver is a major site of dengue pathology. In this project, transendothelial electrical resistance readings of co-cultures of endothelial cells with either SVPs or HSCs both revealed the DENV serotypes to significantly lower endothelial membrane function, most notably at DENV-2. Additionally, preliminary data from the 3D Matrigel angiogenesis assay revealed the DENV serotypes to alter the ability of endothelial cells and SVPs to form vascular structures. Interestingly, our alamar blue proliferation assay utilising hepatic stellate cells (HSCs) showed that liver pericytes display increased proliferation in response to the various DENV serotypes, with immunostaining of HSCs also revealing an increase in myofibroblast protein alpha-smooth muscle actin. These results collectively show the ability to disrupt endothelial-pericyte interactions is shared across the DENV serotypes and suggest a potential loss of endothelial-pericyte interaction due to pericyte differentiation in the liver. This information gives further insight to the mechanisms related with DENV interactions with the vasculature, which could aid in establishing a diagnostic test or treatment for those affected with severe dengue fever.
Myostatin deletion reverses myosteatosis and improves angiogenesis in obese mice
Authors: Speese A , Padgett C A , Rosewater C L , Corley Z L , Mintz J D , Fulton D J , Stepp D W 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): T32 Training Grant: Multi-Disciplinary Training Program in the Mechanisms of Cardiometabolic Disease

Objective
To test the hypothesis that increased muscle mass achieved through deletion of myostatin improves ischemic angiogenesis in obese mice.

Methods
db/db mice, a well-described model of obesity, were crossed with mice lacking myostatin (MSTN KO), a myokine that negatively regulates muscle differentiation and growth, to generate lean and obese mice with and without elevated muscle mass (MSTN KO). We utilized confocal and electron microscopy (EM) and nuclear magnetic resonance (NMR) spectroscopy to assess morphology and histology in the skeletal muscle (SKM) of db +/-_MSTN +/- mice. Muscle lipid contents were assessed via mass spectrometry. Femoral artery ligation was used to stimulate in vivo angiogenesis in response to ischemia. Gene expression was assessed by RNA Seq with mRNA and protein expression follow-up in isolated gastrocnemius skeletal muscle via RT-qPCR and western blotting.

Results
As previously described, myostatin deletion results in significantly increased muscle mass without altering whole-body mass, fat percentage, or activity levels in obese db/db mice. Blood glucose is markedly improved but plasma lipids remain elevated. In parallel with these improvements, we observed that obesity inhibits and MSTN deletion restores vascularization following hind limb ischemia. Despite persistent plasma lipemia, ectopic lipid deposition in skeletal muscle was largely ameliorated in obese mice lacking myostatin, a reduction largely explained by a decrease in tissue triglyceride levels. EM and confocal microscopy revealed accumulation of lipid in obese mouse muscle cells that was lacking in obese mice with MSTN KO. The resolved deposition occurred predominantly inside muscle cells with fat cell infiltration in external spaces largely unaffected. RNA Seq analysis of skeletal muscle revealed marked upregulation of lipogenesis pathways, notably the expression of stearoyl-CoA desaturase-1 (SCD1), the rate limiting enzyme for the oleate synthesis and subsequent triglyceride production. SCD1 was elevated in muscle from normal obese mice but not those lacking myostatin at both the protein and RNA level.

Conclusion
In summary, increased muscle mass in obese mice provides potent protection to vascular and metabolic health. A potent correlate of this improvement is the resolution of myosteatosis in obese, hypermuscular mice. These data suggest that resolution of myosteatosis independently of muscle mass may afford similar protection, possibly by targeted deletion or blockade of SCD1.
Increased oxidative stress alters coronary microvascular tone in exercising swine with multiple comorbidities
Authors: Sorop O , Van Drie RWA , Van De Wouw J , Duncker DJ , Merkus D 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Foundation. Main funding source(s): Netherlands CardioVascular Research Initiative: an initiative with support of the Dutch Heart Foundation

Introduction
Diabetes mellitus (DM), chronic kidney disease (CKD) and hypercholesterolemia either alone or in combination, induce sustained systemic inflammation, increased oxidative stress and coronary endothelial dysfunction. The resulting alterations in microvascular tone may contribute to impaired myocardial perfusion in patients chronically affected by these comorbidities.

Purpose
We tested, in exercising swine, the hypothesis that increased production of reactive oxygen species (ROS) due to DM, CKD and high fat diet (HFD) results in loss of NO bioavailability in the coronary microvasculature leading to altered myocardial perfusion.

Methods
In 12 female swine, DM (streptozotocin 3x50mg/kg iv), CKD (renal embolization), and hypercholesterolemia (via high fat diet, HFD) were induced for 6 months (DM+HFD+CKD). 10 female healthy swine on normal pig chow served as controls (Normal). The role of ROS in the regulation of coronary flow was studied at rest and during graded treadmill exercise with or without ROS scavengers (MPG+Tempol).
Results 6 months after the induction of comorbidities, DM+HFD+CKD animals had sustained hyperglycemia (19.5±1.1 vs 8.7±0.5mmol/L), renal dysfunction (plasma creatinine: 163±7 vs 120±3 µmol/l), hypercholesterolemia (12.7±2.1 vs 1.7±0.1mmol/l) and sustained systemic inflammation (TNF 52±5 vs 25±6 pg/ml, all P&lt;0.05). Myocardial superoxide production was increased in DM+HFD+CKD animals due to NADPH oxidase activation and eNOS uncoupling, (Sorop et al, CVR 2018) and total antioxidant capacity was reduced (Fig. A), which was associated with a lower myocardial NO production, (P&lt;0.05). In vivo experiments showed that myocardial oxygen delivery was impaired in DM+HFD+CKD swine at rest and during exercise, requiring an increased myocardial oxygen extraction (MO2ex) compared to Healthy (compare control runs in panels C and D, P&lt;0.05), that is due to a loss of NO vasodilator influence (van de Wouw et al. BRIC 2021). Interestingly, scavenging of ROS had no effect in Healthy but resulted in increased MO2ex in DM+HFD+CKD (Fig. C, D), indicating that increased oxidative stress resulted in increased production of vasodilator ROS, most likely H2O2. The latter was also supported by an increase in ceramide production (138±34 vs 45±4 nmol/ml, P&lt;0.05) and increased activity of catalase in the myocardium (Fig. B, P&lt;0.05).

Conclusion
In swine, 6 months exposure to multiple comorbidities resulted in increased oxidative stress associated with impaired myocardial oxygen delivery, due to a loss of NO that was partially compensated for by increased H2O2-mediated coronary vasodilation.
Changes in endothelial characteristics as a new strategy to identify intracranial aneurysm at risk of rupture
Authors: Cayron A , Morel S , Bijlenga P , Allemann E , Kwak B 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: None.

Background
Intracranial aneurysm (IA) is an arterial disease resulting in abnormal enlargement of the vessel lumen. Most IAs are asymptomatic but their rupture induces subarachnoid hemorrhage with 24% resulting in death and 11% in severe disability. Occlusion of an unruptured IA by microsurgical or endovascular treatment results in a procedural morbidity and mortality of between 4 to 7%. Decision making between "intervention" or "continuing observation" for an individual patient is currently based on clinical scores and further supported by radiologic imaging of IA characteristics including the observation of vessel wall enhancement (VWE) on magnetic resonance imaging (MRI). VWE reflects a local accumulation of colloid contrast agent, which has been associated with IA wall instability. The pathophysiology of VWE is unknown but enhanced permeability of arterial endothelium, excessive macrophage infiltration, or presence of vasa vasorum have been proposed as potential mechanisms.

Purpose
The goal of our study is to investigate the expression of tight junction (TJ) proteins in the arterial endothelium of human IA domes in the context of IA wall instability. To this end, we have compared ruptured and unruptured IA domes from a human IA biobank.

Methods
Human aneurysms from the middle cerebral artery were obtained during microsurgery by resection of the IA dome after clipping of the neck, embedded in paraffin, and then sectioned and stained for the endothelial cell marker CD31 and the TJ proteins Claudin-5 and Zonula occludens-1 (ZO-1). The expression of these proteins was compared between ruptured and unruptured IAs. Results are expressed as a median (interquartile range).

Results
Histological analyses for CD31 revealed that endothelial cell coverage in ruptured (N=16) IAs tended to be decreased compared to unruptured (N=30) domes (27% (10-49) versus 38% (23-77) respectively, P=0.06). Interestingly, in ruptured aneurysms the expression of the TJ proteins Claudin-5 (1.70um (1.52-2.04)) and ZO-1 (2.29um (1.96-2.75) was lower in comparison with unruptured samples (Claudin-5: 2.15um (1.90-2.76), P&lt;0.05; ZO-1: 2.66um (2.40-3.66), P&lt;0.05).

Conclusion
Our study revealed reduced expression of Claudin-5 and ZO-1 in ruptured IAs in comparison with unruptured IA domes. Reduction in TJ proteins may lead to increased endothelial permeability thus favoring the accumulation of MRI contrast agent in the IA wall and the observation of VWE. Focusing on changes in endothelial characteristics and permeability may represent a new strategy to identify IAs at risk of rupture.
Saraf implication in the vascular remodeling
Authors: Martin Bornez M , Rosado JA , Ordonez A , Smani T 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public Institution(s). Main funding source(s): Agencia Estatal de Investigación

Background
Orai1 and STIM1, molecular components of Store-Operated Calcium Entry (SOCE), have been associated with vascular smooth muscle cells (VSMCs) proliferation in vascular remodeling. Nevertheless, the role of SARAF (SOCE Associated Regulatory Factor), a regulatory protein involved in STIM1 inhibition, has not been firmly established in the vascular remodeling.

Objetive
The aim of this study was to examine the role of SARAF and Orai1 in VSMCs proliferation and neointima formation after balloon injury of rat carotid arteries.

Methods and Results
Experiments were conducted in an animal model of rat carotid angioplasty to characterize neointima formation. VSMC isolated from rat coronary artery was also used to examine cell proliferation. The formation of neointima after balloon injury of rat carotid arteries was confirmed by haematoxylin and eosin staining of tissue sections up to 3 weeks after surgery. Injured arteries showed significant higher expression of SARAF, STIM1 and Orai1 compared to control tissues, corroborating the presence of these regulatory proteins in the neointima layer. Proximity ligation and co-immunoprecipitation assays revealed that SARAF interacts with Orai1 in the neointima. Furthermore, selective silencing of SARAF and Orai1 by small interfering RNA (siRNA) inhibited VSMC proliferation.

Conclusions
Our data suggest that SARAF is involved in VSMC proliferation and neointima formation after vascular injury.
Multiplexed apolipoprotein profiling advances the assessment of residual lipid-related cardiovascular risk
Authors: Schmidt L , Clarke R , Von Ende A , Yin X , Theofilatos K , Hopewell J , Mayr M 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Foundation. Main funding source(s): British Heart Foundation

Background and Purpose
Low-density lipoprotein cholesterol (LDL-C) is among the conventional lipid parameters used to predict risk of cardiovascular disease (CVD). Statins lower blood levels of pro-atherogenic LDL-C, but a residual cardiovascular risk remains in some individuals with therapeutically optimised LDL-C levels. Although the metabolism of LDL-C and other lipoprotein particles is governed by a range of different apolipoproteins, only apolipoproteins A-I and B are measured in clinical assays. Using a more comprehensive apolipoprotein panel in a large epidemiological cohort, this study aimed to determine the association of individual apolipoprotein levels with risk of coronary heart disease (CHD).

Methods
Bottom-up multiple reaction monitoring–mass spectrometry (MRM–MS) was used in conjunction with stable isotope-labelled peptide standards to quantify plasma levels of 13 apolipoproteins in participants of the Precocious Coronary Artery Disease (PROCARDIS) study (N = 1916; 941 cases of CHD, 975 controls). The relationship between apolipoprotein levels and CHD was assessed after adjusting for established risk factors for CVD and correcting for statin use.

Results
The strongest positive associations with CHD in the PROCARDIS study were seen for triglyceride-related apolipoproteins C-I (odds ratio [OR] 2.38, 95% confidence interval [CI] 1.63–3.46), C-III (OR 2.95, 95% CI 1.85–4.71) and E (OR 2.35, 95% CI 1.54–3.58), as well as for apolipoprotein (a) (kringle IV type 2 repeat, OR 2.84, 95% CI 2.04–3.95). Comparing these with associations of apolipoproteins with CVD in the Bruneck study (N = 688) revealed consistency across the two cohorts. Robust inverse associations with CHD were observed for apolipoproteins A-IV (OR 0.45, 95% CI 0.31–0.65) and M (OR 0.29, 95% CI 0.19–0.44).

Conclusion
Analysing two large epidemiological cohorts, Bruneck and PROCARDIS, demonstrated that multiplexed apolipoprotein profiling improves the understanding of cardiovascular risk independent of conventional lipid parameters. Most prominently, triglyceride-related apolipoproteins were shown to positively associate with residual cardiovascular risk. The findings of this study support the need for development and implementation of standardised, MRM–MS-based apolipoprotein profiling assays to guide novel lipid-modifying therapies beyond statins.