bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bradycardic effect of endothelial-specific bradykinin type-II receptors overexpression in mice: impact on blood pressure
Authors: Metry S , Suvorava T , Kojda G 
Published: November 2023

Abstract

Background/Introduction
Polymorphisms of the gene encoding for the bradykinin type-II receptors (B2R) has been shown to decrease the expression of B2R and increased vascular resistance and blood pressure in humans.

Purpose
We aimed to study the role of endothelial B2R on systolic blood pressure (SBP) and heart rate (HR) using a novel mice model with endothelial-specific B2R overexpression (B2tg) as compared with transgene negative littermates (B2n) and on differential expression of proteins potentially relevant to vascular tension.

Methods
Human and murine B2R expression levels were quantified by rtPCR. Thoracic aortic tissues were used for vascular reactivity studies and proteomic analysis. SBP and HR were measured by the tail-cuff method at baseline, after administration of the COX inhibitor diclofenac (10 mg/kg/day), B2R antagonist icatibant (1 mg/kg) or NOS inhibitor L-NAME (100 mg/kg/day).

Results
In aortic tissues of B2tg human B2R were exclusively expressed (n=6, P=0.0005), while murine B2R expression levels remained unchanged. Similar results were found in skeletal muscle tissues used as an estimate for resistance vessels. Aortic ring studies revealed a concentration-dependent constriction to bradykinin in wild-type mice (maximal vasoconstriction 9.6±3.0%, n=5) whereas aortic rings of B2tg showed concentration-dependent relaxation (-27.4±4.1%, n=12). SBP was significantly lower in B2tg (125.0 ±1.6 mmHg) as compared to B2n (129.9±1.1 mmHg, n=9, P=0.03). Diclofenac treatment slightly but not significantly reduced SBP in B2n (to 125.5±2.4 mmHg, P=0.09) and in B2tg (to 119.8±2.8 mmHg, n=9, P=0.12), however the difference in SBP remained (P=0.03). In a pilot study, L-NAME administration did also not change the SBP difference between B2n and B2tg (9.9±10.3 mmHg, n=3-4). In contrast, icatibant administration abolished the difference between the groups (B2n 128.6±4.6 vs B2tg 128.3±2.0 mmHg, n=8, P=ns) suggesting the involvement of endothelial B2R in SBP reduction. Interestingly, in the same mice, HR was significantly lower in B2tg (571.4±20.8 bpm) as compared to B2n (626.7±13.7 bpm, P=0.02). While diclofenac didn’t affect HR, icatibant significantly increased HR to (630.8±21.8 bpm in B2tg and 685.5±16.4 bpm in B2n, n=8, P=0.01). Only in B2tg, there was a significant strong correlation between SBP and HR (r=0.76, P=0.016). Analysis of proteomic data revealed differential upregulation of proteins related to cell adhesion (CD44), extracellular matrix (hyaluronidase), and differential downregulation of proteins related to oxidative stress (superoxide dismutase) in B2tg.

Conclusion
Endothelial B2R overexpression reduces SBP and HR. SBP reduction is not dependent on two major vascular pathways stimulated by products of NOS and COX but appears to be related to endothelial B2R-dependent HR reduction. Hence, agonism at B2R could be a valuable target for development of new cardiovascular drugs.
B cells and immunoglobulins promotes aortic dissection in mice
Authors: Ohno S , Aoki H , Furusho A , Hashimoto Y , Nakao E , Shibata R , Nishida N , Ito S , Majima R , Fukumoto Y 
Published: November 2023

Abstract

Background
Aortic dissection (AD) is a devastating aortic disease with high mortality, for which molecular pathogenesis is largely unknown. Recent studies have highlighted the importance of inflammatory response in AD. We have reported that B cells and immunoglobulins promote inflammatory response in the aorta and are involved in the pathogenesis of abdominal aortic aneurysm, another form of aortic disease. However, it is not known whether and how B cells participate in AD development or progression.

Methods and results
Immunohistochemical staining of human AD tissue revealed that B cells were clustered together with T cells, macrophages, and neutrophils at the entry site of AD with medial disruption. B cell cluster was also observed at the site of medial disruption in mouse model of AD that was induced by continuous infusion of beta-aminopropionitrile and angiotensin II (BAPN+AngII). In muMT mouse, which is genetically deficient for B cells and immunoglobulins, BAPN+AngII induced significantly less severe AD compared to that in wild type. The rate of aortic rupture and sudden death was approximately 42% in wild type mice, while that in muMT mouse was 12% (P&lt;0.05). Administration of mouse normal polyclonal IgG to muMT mice resulted in dramatic increase in aortic rupture and sudden death, starting at day 7 of BAPN+AngII infusion, and reaching 69% of rupture rate, indicating the critical role of IgG in AD. Transcriptome analysis revealed that IgG administration enhanced the expression of inflammation-related genes in muMT mice before BAPN+AngII infusion, indicating that IgG increased susceptibility to AD. Immunohistochemistry of aorta confirmed exogenous IgG deposition, colocalized with C3, which is known to contribute to innate immunity.

Conclusion
These findings demonstrated B cells and IgG are critically involved in the destructive inflammation of AD pathogenesis. Further, the aortic deposition of IgG and C3, a part of the components of innate immunity, precedes the development of AD. These findings may provide the therapeutic opportunities to AD.
Proteomic characterization of peripheral leukocytes in variant carriers of CDKAL1 associated with cholesterol efflux capacity
Authors: Lee S H , Ann S J , Choi D 
Published: November 2023

Abstract

Background
Genetic variations near CDKAL1 (CDK5 regulatory subunit-associated protein 1-like 1) gene recently showed association with individual’s cholesterol efflux capacity. The aim of this study was to compare the proteomes of peripheral leukocytes in CDKAL1 variant carriers and controls to identify evidence of biological impact caused by these variants.

Methods
Peripheral blood leukocytes were isolated from five individuals with any of four CDKAL1 variants (rs117835232, rs117252933, rs118064592, and rs150434350) and five controls. Cell lysates were digested to peptides and Q Exactive Orbitrap Plus mass spectrometry was applied for comparative proteomics analysis. Protein identification and quantification were performed with MaxQuant software.

Results
In total, 2,387 proteins were identified at false discovery rate 1% level, of which 176 proteins were differently abundant between CDKAL1 variant and control groups (p &lt;0.05, fold-difference &gt;1.5). Among them, 39 upregulated and 137 downregulated proteins were identified in the variant group (p &lt;0.05; fold change&gt;1.5) (Figure 1). The upregulated included docosahexaenoic acid omega hydroxylase, resistin, and apoA1, whereas the downregulated included NADH ubiquinone oxidoreductase. The functional enrichment and KEGG pathway analysis showed 20 associated pathways including carbon metabolism, synthesis of antibiotics, and RNA transport.

Conclusions
Differential expression of proteins regulating lipid metabolism and RNA transport in CDKAL1 variant carriers indicates differences in corresponding biological function in these individuals. Detailed contribution of the reported proteins remains to be clarified.
Cyclodextrin-loaded nanobubbles reduce cholesterol and atherosclerosis in vivo
Authors: Wang X , Mehta S , Bongcaron V , Nguyen T , Jirwankar Y , Maluenda A , Walsh A P G , Palasubramaniam J , Hulett M , Srivastava R , Bobik A , Peter K 
Published: November 2023

Abstract

Background
Ischemic heart disease (IHD) is the leading cause of mortality and morbidity worldwide. Atherosclerosis, a chronic inflammatory disease, is the main contributor to IHD. However, atherosclerosis is commonly left undiagnosed and untreated. Our study aims to create a theranostic nanobubbles (NBs), with ultrasonic properties for diagnostic imaging and at the same time for the delivery of 2-Hydroxypropyl-beta-cyclodextrin (CD), an FDA-approved drug to dissolve cholesterol crystals.

Methods and Results
Using polybutlycyanoacrylate, we created polymeric NBs. The incorporation of near-infrared dye (IR780) resulted in a multimodal contrast NB, which allows in vitro and in vivo visualization on both ultrasound and optical imaging. Furthermore, these NBs can be triggered by ultrasonic bursts to release their dye/drug payload. The safety profile of the NBs, post-incorporation of CD (CDNBs), demonstrated excellent biocompatibility in vitro using cell culture and in human blood. Cellular uptake of CP increased by 2.4x when using CPNBs compared to free CD (p&lt;0.00001). In vivo biodistribution demonstrated that the NBs circulate in blood plasma for 48 hours, with a peak at 1-hour post-injection. In a mouse model of atherosclerosis, ApoE-/- mice on high-fat diet, we intravenously injected CDNBs, free CD or NBs only fortnightly for 8 weeks. En face Oil Red O-staining showed a significant reduction in total plaque area in the CDNB-treated group, as compared to the control animals treated with free CD and with NBs only (p&gt;0.01). There was also a significant reduction in blood plasma cholesterol for the CDNBs-treated mice as compared to the control mice (p&gt;0.01).

Conclusion
We demonstrated the ability to load CDs into NBs. These CDNBs are taken up by immune cells. The passive accumulation of CDNPs in the atherosclerotic lesions, resulted in the release of CD and the reduction of total plaque area in the entire aorta. These CDNBs treated mice also have a reduced cholesterol level in their plasma. CDNBs represent a novel theranostic nanocarrier with a high potential of clinical translation and ultimately major benefits to patients with atherosclerosis.
The effect of antibody-mediated PCSK9 inhibitor on bare-metal stent implantation outcome
Authors: Puspitasari Y M , Ministrini S , Liberale L , Vukolic A , Baumann-Zumstein P , Holy E W , Montecucco F , Luscher T F , Camici G G 
Published: November 2023

Abstract

Introduction
Percutaneous coronary intervention (PCI) is the gold standard for the management of acute coronary syndrome, and has been benefited from advances in pharmacotherapy and device innovation. Nevertheless, in-stent restenosis and stent thrombosis remain serious complications following PCI procedure with stent implantation. Proprotein convertase subtilisin kexin type 9 (PCSK9) is an enzyme known for its role in plasma cholesterol homeostasis. Efficiently reducing plasma cholesterol, PCSK9 inhibition recently became a pivotal treatment strategy for hypercholesterolemia. However, the effect of PCSK9 inhibition on stent implantation outcomes remains uninvestigated.

Methods
Carotid artery bare-metal stent (BMS) implantation was performed on C57Bl/6 mice, which then a parallel assessment of various inflammatory mediators was performed. To inhibit PCSK9, alirocumab was administered weekly to antiplatelet-treated mice started before stent implantation. Six weeks after the stent implantation procedure, animals were harvested and histomorphometric analysis was performed on stented arteries. In vitro cell migration and cellular senescence were also assessed on alirocumab-treated primary human aortic endothelial (EC) and vascular smooth muscle cells (VSMC).

Results
Compared to sham intervention, stent implantation was associated with increased expression of several inflammatory mediators, including PCSK9. Interestingly, the increase in PCSK9 level was confirmed in the stented vascular tissue, but not in plasma (Fig 1A). Following alirocumab treatment, PCSK9 inhibition results in an increased intimal hyperplasia in the stented vascular segment of alirocumab-treated animals compared to controls (Fig 1B). In vitro, alirocumab promoted migration and inhibited the onset of senescence in primary human VSMC, while blunted the migration and increased the senescence of EC.

Conclusion
The results of this study demonstrated that antibody-based PCSK9 inhibition worsen bare-metal stent implantation outcome in mice. This observed effect is mediated, in part, by the differential effect on VSMC and EC senescence, thus promoting in-stent intimal hyperplasia while blunting vascular healing. The herein-reported data warrant additional investigations concerning the use of PCSK9 inhibitors in patients undergoing PCI with stent implantation.
Phenylacetylglutamine causes a pathologic inflammation state and enhances atherosclerosis through the b2-adrenergic receptor cAMP PKA NF-kappaB pathway in diabetes
Authors: Huang L 
Published: November 2023

Abstract

Background
Type 2 diabetes (T2DM) affects billions of patients and has a pathophysiologically important inflammatory component. Macrophage plasticity in diabetes is impaired, and remains in a persistent proinflammatory state; yet currently, the cause of the inflammatory state is unknown. Phenylacetylglutamine (PAGln) is an amino acid metabolism end product that rises in T2DM patients and is associated with incident major adverse cardiovascular events. However, the role of PAGln in the inflammatory component of T2DM remains unknown.

Purpose
Here, we aim to identify blood-based systemic factors for pathologic inflammation status and risk of atherosclerosis as they are highly reproducible at low cost and search for effective and safe agents to attenuate diabetic inflammatory complications.

Methods
In vivo, phenylacetylglutamine was dissolved in normal saline and injected in mice daily (intraperitoneal). The levels of PAGln were analyzed by liquid chromatography-mass spectrometry. The effect of PAGln on atherosclerosis progression was analyzed by Ldlr–/– mice, followed by a Western diet feeding for 12 weeks. In vitro, primary mice bone marrow-derived macrophages (BMDMs) were treated with phenylacetylglutamine (100 uM) for the indicated time. Elisa, PCR, and western blot were used to characterize the impact of PAGln on BMDMs and mice arteries. Flow cytometry was taken to quantify and type immune cells in mice bone marrow, blood, spleens, and arteries. The activity of the NF-κB pathway was assessed by KEGG analysis, western blot, luciferase, and activity assay.

Results
In vivo, increased inflammatory macrophages were detected in mice blood, spleens, bone marrow, and arteries after PAGln injection. In vitro, PAGln leads to macrophage M1 polarization. Taken together, PAGln disturbed the transition of macrophage phenotype from inflammatory to reparative, facilitating pathologic inflammation. The RNA of BMDMs after PAGln treatment was collected for RNA-seq. KEGG analysis and western blot prove PAGln exerted its function through the activation of NF-κB. PAGln could promote NF-κB signals by increasing the phosphorylation level of p65. Besides, the negative effects of PAGln could be rescued by ß-blockers.

Conclusion
Our study unravels a novel mechanism by which PAGln accelerates atherosclerosis progression and causes pathologic inflammation by activating the ß2-adrenergic receptor /cAMP/PKA/NF-κB pathway. The negative effect of PAGln could be rescued by carvedilol (ß blocker). Thus, our recognition of chronic inflammation in diabetes needs to be revised, and understanding these mechanisms can inform alternative treatment strategies and refine atherosclerosis repair therapies for diabetic patients.
Synergistic effect of early rhythm control and healthy lifestyle on the risk of dementia in patients with new-onset atrial fibrillation
Authors: Kim Y , Lee S R , Choi E K , Lee S W , Han K D , Oh S , Lip G 
Published: November 2023

Abstract

Background
Early rhythm control (ERC) and healthy lifestyle (HLS) have been found to be related to lower dementia risk in patients with atrial fibrillation (AF), however, it has not been evaluated whether there is a synergistic effect between the two.

Methods
Using the Korean National Health Insurance Service database, patients who were newly diagnosed with AF between 2009 and 2016. Patients were categorized as follows: those who did not receive ERC including antiarrhythmic drug, cardioversion, AF catheter ablation within 2-year after AF diagnosis, and had 1 or more unhealthy lifestyle behaviors among current drinking, current smoking, and lack of regular exercise (group 1), those with HLS, but not receiving ERC (group 2), those receiving ERC, but with at least 1 unhealthy lifestyle behavior (group 3), and those with HLS and receiving ERC (group 4). To compare 4 groups, propensity score weighting method was used to balance the baseline characteristics of 4 groups. During follow-up, incident dementia, vascular dementia, and Alzheimer’s dementia were evaluated.

Results
A total of 199,947 patients were included. The mean time from AF diagnosis to implementation of ERC of groups 3 and 4 was 27±77 and 27±74 days, respectively. Among total study population, 96,907, 53,293, 29,234, and 20,513 patients were included in group 1, 2, 3, and 4, respectively. Group 4 showed the greatest risk reduction for all dementia, vascular dementia, and Alzheimer’s dementia among 4 groups (Figure). Both ERC alone and HLS alone were associated with a lower risk of vascular dementia by 20% than group 1. The combination of the two was associated with a lower risk of vascular dementia by 40% (Figure). However, the impact of ERC on the risk reduction of Alzheimer’s dementia was statistically insignificant, and maintaining HLS seemed sufficient to lower the risk of Alzheimer’s dementia (Figure).

Conclusion
The combination of ERC and HLS might be beneficial to prevent vascular dementia in patients with new-onset AF. To reduce the risk of Alzheimer’s dementia, maintaining HLS should be emphasized.
Vascular endothelial dysfunction induced by 3 bromofluoranthene via MAPK mediated NF B proinflammatory pathway and intracellular ROS generation
Authors: Su C H 
Published: November 2023

Abstract
3-Bromofluoranthene (3-BrFlu) is the secondary metabolite of fluoranthene, which is classified as a polycyclic aromatic hydrocarbon, through bromination and exists in the fine particulate matter of air pollutants. Endothelial dysfunction plays a critical role in the pathogenesis of cardiovascular and vascular diseases. Little is known about the molecular mechanism of 3-BrFlu on endothelial dysfunction in vivo and in vitro assay. In the present study, 3-BrFlu included concentration-dependent changes in ectopic angiogenesis of the sub-intestinal vein and dilation of the dorsal aorta in zebrafish. Disruption of vascular endothelial integrity and up-regulation of vascular endothelial permeability were also induced by 3-BrFlu in a concentration-dependent manner through pro-inflammatory responses in vascular endothelial cells, namely SVEC4-10 cells. Generation of pro-inflammatory mediator PGE2 was induced by 3-BrFlu through COX2 expression. Expression of COX2 and generation of pro-inflammatory cytokines, including TNFa and IL-6, were induced by 3-BrFlu through phosphorylation of NF-kB p65, which was mediated by phosphorylation of MAPK including p38 MAPK, ERK and JNK. Furthermore, generation of intracellular ROS was induced by 3-BrFlu, which is associated with the down-regulated activities of the antioxidant enzyme (AOE), including SOD and catalase. We also found that 3-BrFlu up-regulated expression of the AOE and HO-1 induced by 3-BrFlu through Nrf-2 expression. However, the 3-BrFlu-induced upregulation of AOE and HO-1 expression could not be revised the responses of vascular endothelial dysfunction. In conclusion, 3-BrFlu is a hazardous substance that results in vascular endothelial dysfunction through the MAPK-mediated-NFkB pro-inflammatory pathway and intracellular ROS generation.
PCSK9-inhibitor therapy leads to changes in immune cell gene expression and function: A subset analysis of the PACMAN-AMI trial
Authors: Ondracek A , Hofbauer T M , Aszlan A , Kuehn S , Seidl V , Toma A , Speidl W , Hengstenberg C , Raber L , Lang I M 
Published: November 2023

Abstract

Introduction
The PACMAN-AMI trial showed a significant reduction of plaque atheroma volume by alirocumab on top of high-intensity statin therapy as compared to placebo in 265 patients undergoing serial multi-vessel imaging after acute myocardial infarction (AMI). Neutrophils and the formation of neutrophil extracellular traps (NETs) have been implicated in the development and progression of atherosclerosis. High levels of low-density lipoprotein (LDL) have been associated with neutrophil activation.

Objectives
This study aimed to determine the influence of aggressive LDL-lowering on neutrophil activation, and immune cell gene expression of pro-inflammatory markers in a subset of the PACMAN-AMI cohort.

Methods
Neutrophils and peripheral blood mononuclear cells (PBMC) of 38 patients were isolated by serial density-gradient centrifugation to assess relative gene expression of IL-1b, IL-6, TNFa, X-box binding protein (XBP) 1, caspase 1 (CASP1), and NLRP3 by quantitative PCR. Neutrophils were stimulated to undergo NET formation using a calcium ionophore in the presence or absence of pro-protein convertase subtilisin/kexin type 9 (PCSK9), detecting the release of double-stranded (ds)DNA. Plasma samples were screened for dsDNA, citrullinated histone H3, neutrophil elastase, and myeloperoxidase. Healthy controls (n=30) were recruited during routine check-ups in a health and prevention center.

Results
After one year of double-blind injections, the probability to undergo NET formation in vitro was still higher in AMI patients than in healthy controls. Considering randomized treatment, only patients injected with alirocumab showed significantly higher NET release than controls. Additional stimulation with the alirocumab target PCSK9 could reduce NET formation in all cohorts. This effect was significantly higher in patients treated with alirocumab. Relative gene expression of CASP1, IL-1b, IL-6, and XBP 1 was higher in patients treated with alirocumab. There was no difference in plasma NET markers comparing healthy controls and both treatment arms.

Conclusion
Our data indicate that PCSK9 might have effects beyond its role in cholesterol metabolism influencing immune cell activation, which could mask effects of LDL lowering. Treatment does not directly translate to higher plasma NET levels without an additional inflammatory stimulus disrupting the steady state.