bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 208: Laminar Shear Stress Decreases Endothelial Microparticles Release by a Nitric Oxide- and ABCA1-Dependent Mechanism
Authors: Vion Anne C , Ramkhelawon Bhama , Loyer Xavier , Chironi Gilles , Tedgui Alain , Lehoux Stephanie , Boulanger Chantal M 
Published: March 2022

Microparticles (MP) are shed-membrane sub-micron vesicles released upon cell activation or apoptosis. Their formation is associated with loss of cell membrane asymmetry and exposure of phosphatidylserine (PS), resulting from increases in floppase activities such as ABCA1. Plasma endothelial MP (EMP) are surrogate markers of endothelial dysfunction and their levels increase in patients with cardiovascular diseases. However, the mechanisms governing EMP release in vivo remain largely unknown. As nitric oxide (NO) is a key regulator of the vasoprotective effect of laminar shear stress (SS) on the endothelium, we tested the hypothesis that endothelial NO regulates EMP release under different SS profiles.
Shed-annexinV+ EMPs were analyzed by flow cytometry in the medium of confluent HUVECs (passage 1-3) subjected to either high- (15 dyne/cm2; atheroprotective) or low- (1.5 dyne/cm2; atheroprone) SS conditions for 24 hrs. HUVECs exposed to high-SS released 3-fold less EMPs compared to low-SS conditions (326±37 vs. 130±23AnnV+MPs/μL, p<0.001). High-SS EMPs also externalized less PS, as estimated by the mean fluorescence of FITC-AnnV labelling (15.6±6.1 vs. 24.2±9.4 AU, p<0.001). Exposure to L-NAME (10-5M), but not D-NAME, increased high-SS-induced EMP release by over 3-fold (p<0.001) and the NO donor SNAP (10-5M) significantly decreased low-SS-induced EMP formation by 50% (p=0.05). In addition, L-NAME increased the mean fluorescence of EMP labelling under high SS (p<0.05), whereas SNAP decreased it under low SS (p<0.05). We then evaluated ABCA1 expression under these conditions. high-SS significantly decreased the expression of ABCA1 both at the mRNA and protein level by 2- and 20-fold, respectively (p<0.05 vs low SS). L-NAME augmented ABCA1 mRNA level in cells exposed to high SS (p<0.05), whereas SNAP significantly decreased ABCA1 mRNA level under low SS conditions (p<0.05). Finally, the LXRα agonist TO90131 (0.1mM) not only increased endothelial ABCA1 expression by 20-fold, but also augmented MP level under high SS conditions.
Altogether, these results demonstrate that atheroprotective shear stress limits the release of procoagulant endothelial MP through a NO-dependent regulation of endothelial ABCA1 expression.
Abstract 29: Fate-Tracing Demonstrates that Medial SMCs, but Not Mature ECs, Contribute to Neointimal Cell Population in Injured Arteries
Authors: Hong Michael , Gupta Tushar , Fu Chunhua , Berceli Scott , Jiang Zhihua 
Published: March 2022

It has been proposed that various groups of cells, such as medial SMCs (mSMCs), bone marrow progenitors, and mature ECs (mECs) hold the potential to become neointimal SMCs via phenotypic switching, differentiation, or endothelial-mesenchymal-transition (EMT). However, no definitive evidence has been provided to support these claims, which renders the in vivo significance of these events in question.
Using an inducible Cre-loxP system, where the production of a reporter (e.g. β-gal) is driven by a highly definitive marker gene for mSMCs (e.g. SMC myosin heavy chain, SMMHC) or mECs (e.g. stem cell leukemia, SCL), we traced the fate of these cells during neointimal hyperplasia (NIH) in ligated common carotid and wire-injured femoral arteries. Tamoxifen induction resulted in a homogeneous labeling of mSMCs of the unmanipulated arteries of R26R;SMMHC-CreERtm mice. Following arterial injury, SMCs that were initially located in the medial layer presented in neointimal lesions; however, the fraction of these cells varied widely among vessels with either type of injury. This heterogeneity was not secondary to the random loss of the genetic label as evidenced by the stable β-gal activity in SMC culture. Addition of a second (4wks after surgery) induction did not change the pattern and fraction of the labeled SMCs in the neointima, indicating that few recruited progenitors transform into mature SMCs. This observation is further supported through examination of mice chimeric for R26R;SMMHC-CreERtm bone marrow, where β-gal activity was not detected in injured arteries though strongly induced in culture following PDGF treatment. Neointima formed in R26R;SCL-CreERtm arteries demonstrated a co-localization of β-gal activity to the repopulated ECs, with none being detected in cells underneath the EC monolayer, suggesting a role for mECs in re-endothelialization and indicating EMT as a rare event in NIH development.
We have provided direct evidence for the contribution of mSMCs to NIH. The recruited vascular progenitors, particularly bone marrow cells, do not differentiate into fully mature SMCs in NIH lesion. EMT does not serve as a mechanism for populating neointimal cells following vascular injury while occurring under other vascular disease conditions.
Abstract 291: Health Effects of Northern Contaminants and Alcohol Consumption in a Rodent Model of Human Metabolic and Cardiovascular Diseases
Authors: Florian Maria , Yan Jin , Petrov Ivan , Caldwell Don , Coughlan Melanie C , Laziyan Mahemuti , Li Nanqin , Willmore William , Jin Xiaolei (Dawn) 
Published: March 2022

For the last few decades, Northern populations have been exposed to elevated levels of Northern contaminants (NC) such as polychlorinated biphenyls and heavy metals, mainly through consumption of contaminated fish and marine mammals. Along with this, there has also been an increased prevalence of obesity, hypertension, diabetes, and cardiovascular disease. Although changes in lifestyle and diets have been associated with this prevalence of chronic diseases, it remains to be clarified if exposure to NC also plays a part, and if lifestyles choices such as consumption of alcohol may modulate the effects of NC.
This study explored these questions using a rodent model of human metabolic and cardiovascular diseases.
Obese male JCR rats at 8 weeks of age were acclimatized on a purified AIN93G diet. Animals were treated with 10% alcohol in drinking water or drinking water only for 6 weeks. From the third week of alcohol treatment, animals were orally dosed with a mixture of 23 NC (NCM) at 0 (corn oil), 1.6, or 16 mg/kg BW/day for 4 weeks. During the whole study, body weight and water/alcohol consumption were measured daily. Food consumption was measured weekly. Blood and organs were collected and weighed at the end of the study. Tissue samples were analyzed for hematology and serum NCMs, glucose, insulin, and ethanol levels, as well as clinical biochemistry markers.
Serum NCM levels increased with NCM dose, and were mostly in the range of NCM levels found in Inuit blood. Serum NCM levels were generally lower in the rats treated with alcohol than those without. NCM at 16 mg/kg BW slightly decreased body weight gain and water and food consumption especially in the rats given no alcohol. Regardless of alcohol, this dose of NCM significantly increased absolute and relative liver weight, relative kidney weight, and serum C-reactive protein and ethanol levels. NCM also decreased serum paraoxonase-1, insulin, cholesterol, triglyceride, HDL, and LDL levels.
Our data suggested that NCM at levels relevant to human exposure may alter lipid metabolism and/or transport in the liver, insulin secretion from the pancreas, and circulating cardiovascular health markers.

This research was supported by Canadian Institute of Health Research, Postdoctoral Fellowship No 200910-JHF-214255.
Abstract 510: Gene Set Enrichment Analysis of the Dual PPAR-α/γ Agonists Aleglitazar and Muraglitazar Reveals Distinct Metabolic Signatures in Human Cardiomyocytes
Authors: Wright Matthew B , Ebeling Martin , Steiner Guido , März-Weiss Pia 
Published: March 2022

Dual-acting agonists of the peroxisome proliferator-activated receptors α and γ (PPAR-α/γ) have been pursued to reduce cardiovascular risk in type 2 diabetes. Although core PPAR signaling appears conserved, compound-specific on- and off-target effects contribute to profiles of specific molecules. Here, we used gene set enrichment analysis (GSEA) to compare the effect of dual PPAR-α/γ agonists aleglitazar (Ale) and muraglitazar (Mura).
Primary human cardiomyocytes were incubated with non-cytotoxic concentrations of Ale or Mura (both 1 nM to 25 μM). Whole-genome expression studies were carried out using Illumina microarrays. Principal Component Analysis, hierarchical clustering and GSEA using the publicly available Pathway Commons database identified multiple signaling and metabolic networks significantly regulated by both compounds. Effects of Mura at the higher concentrations substantially diverged from both Ale and low concentrations of Mura, suggesting potential off-target effects. Within the Pathways Commons collection analyzed, four networks were significantly differentially affected by 25 μM Mura versus the other treatments.

Nuclear Factor Y, which recognizes CCAAT elements, was identified as a common signature of Mura-specific effects on the cholesterol biosynthesis pathway (seven genes,
p
= 0.003 for enrichment).

The divergent effects of Ale and Mura on metabolic, energy regulation and interferon signaling pathways may contribute to their unique biological profile.
Abstract 100: Initial Characterization of Oxidant-Resistant Human ApoAI Transgenic Mice
Authors: Berisha Stela Z , Hung Kim T , Brubaker Greg D , Smith Jonathan D 
Published: March 2022

Introduction
HDL functions, such as its cholesterol acceptor and anti-inflammatory activities, are impaired by myeloperoxidase (MPO), which selectively targets and oxidizes HDL and apoAI. We previously found that the 4WF apoAI isoform, in which the 4 Trp residues are substituted with Phe, is equally competent as wild type (WT) human apoAI to interact with lipids and accept cholesterol in an ABCA1-dependent manner, but is resistant to MPO mediated loss of function.

Hypothesis
The oxidant resistant 4WF apoAI isoform will be superior in mediating reverse cholesterol transport than WT human apoAI in vivo under conditions of elevated oxidative stress.

Methods and Results
We generated transgenic mice expressing the 4WF human apoAI (h-apoAI) isoform. A genomic fragment containing the h-apoAI gene was subjected to site directed mutagenesis to convert the Trp to Phe residues, and microinjected into C57BL/6 fertilized oocytes. We selected the line with the highest h-apoAI expression to compare with existing WT h-apoAI transgenics. We assessed plasma apoAI and HDL-C levels in male and female mice. Male mice have significantly higher h-apoAI levels than females for both isoforms (p<0.05). In general, the WT apoAI transgenics have higher plasma h-apoAI levels than the 4WF transgenics; however there is enough overlap that we were able to select male 4WF and WT h-apoAI transgenic mice having matched plasma h-apoAI levels of ~550 mg/dl, which are described below. HDL-C levels were slightly lower in 4WF vs. WT transgenics (157±4 vs 184±8 mg/dl, respectively, N=5 per group, p=0.03). We performed in vivo reverse cholesterol transport (RCT) assay by s.c. injection of cholesterol labeled and loaded bone-marrow macrophages. RCT to the plasma reflected the HDL-C levels and was lower in the 4WF vs. WT transgenic mice (4.5% vs 6.6%, respectively p=0.007). However, RCT to the liver and feces was equivalent in the 4WF and WT transgenics (NS). This suggests that the flux of HDL-C in the 4WF mice may be higher than in the WT apoAI transgenics, leading to equal RCT to the feces and liver despite lower HDL-C levels.

Conclusion
4WF transgenic mice are now breeding with MPO transgenic mice in order to test our hypothesis that this isoform retains more HDL function under oxidative stress conditions.
Abstract 15: The Helix-Loop-Helix Factor Id3 Links Natural Helper Cells to Proliferation of Atheroprotective B-1a Cells
Authors: Perry Heather , Oldham Stephanie , Fahl Shawn , Harmon Daniel , Que Xuchu , Tsimikas Sotirios , Witztum Joseph , Bender Timothy , McNamara Coleen 
Published: March 2022

Natural immunity is emerging as important for atheroprotection. Natural IgM antibodies, such as E06, that recognize oxidative epitopes on LDL or phospholipids prevent atherosclerosis and are produced by the atheroprotective B cell subset, the B-1a B cell. IL5, an important B-1a B cell mitogen, can be produced by the innate lymphoid cell, the natural helper (NH) cell. We previously demonstrated that splenic B cells provide atheroprotection to B cell deficient mice, an effect dependent on the helix-loop-helix transcription factor, Id3. Taken together, these results raise the hypothesis that Id3 may be an important regulator of splenic B-1a B cells and natural immunity in atheroprotection.

Methods and Results

Id3
-/-
Apoe
-/-

mice at 8 weeks old had fewer B-1a cells in the spleen (200672 ±58019, n = 9) compared to

Id3
+/+
Apoe
-/-

mice (511840 ±88811, n = 9) as determined by flow cytometry. Consistent with fewer B-1a B cells, there were lower levels of serum E06 in

Id3
-/-
Apoe
-/-

mice compared to

Id3
+/+
Apoe
-/-

mice (1573 ±300 RLU vs. 2807 ±263 RLU respectively, n = 8 for each group) as assessed by ELISA.

Id3
-/-
Apoe
-/-

mice had decreased B-1a B cell homeostatic proliferation compared to

Id3
+/+
Apoe
-/-

mice (30.0 ±3.3% vs. 54.4 ± 1.2% respectively, n = 4) as measured by
in situ
CFSE. However, B-1a B cell proliferation and number and levels of E06 were unchanged in mice with B cell specific deletion of Id3 compared to controls. To determine if Id3 is required for IL33 induced levels of IL5, IL33 or vehicle control was i.p. injected into

Id3
+/+
Apoe
-/-

mice or

Id3
-/-
Apoe
-/-

mice every 3 days for 7 days.

Id3
-/-
Apoe
-/-

mice had less IL33 induced serum IL5 levels (5.1 ±3.1 pg mL
-1
, n = 5) compared to

Id3
+/+
Apoe
-/-

mice (67.4 ±22.9 pg mL
-1
, n = 4). Indeed, loss of Id3 significantly attenuated IL33 induced IL5 production in NH cells measured by flow cytometric intracellular cytokine staining (

Id3
-/-
Apoe
-/-

: 31.1 ±5.4%, n = 6 vs.

Id3
+/+
Apoe
-/-

: 50.3 ±4.3%, n = 6). NH cells are also present and can to produce IL5 in response to exogenous IL33 (vehicle: 4.0 ±1.4%, n = 4 vs. IL33: 14.7 ±2.4%, n = 4) in the aortic adventitia/perivascular adipose tissue of

Apoe
-/-

mice.

Conclusion
Results provide the first evidence implicating Id3 as a key regulator of NH cell IL5 production and B-1a B cell proliferation.
Abstract 175: Efficacy of Biomimetic Peptide-Fluorosurfactant Polymer Coated Expanded Poly(tetrafluoroethylene) Vascular Grafts in a Porcine Carotid Artery Interposition Model
Authors: Bastijanic Jennifer M , Dudash Lynn A , Kligman Faina , Allemang Matthew T , Lakin Ryan O , Eslahpazir Benjamin A , Kashyap Vikram S , Kottke-Marchant Kandice , Marchant Roger E 
Published: March 2022

Objectives
Vascular grafts currently see limited use in small diameter applications due to the prevalence of thrombosis. Patency of ePTFE grafts could be improved by reducing thrombosis via an endothelial cell specific peptide fluorosurfactant polymer (FSP) coating.

Methods
In this study, ePTFE grafts were coated with cRRE-FSP, RGD-FSP, or Maltose-7 (M7)-FSP, sodded with porcine pulmonary artery endothelial cells (PPAECs), and evaluated in vivo in a porcine carotid artery interposition model. Grafts were excised after one month and evaluated by quantifying thrombosis and intimal hyperplasia and by qualitatively examining inflammation, smooth muscle cell infiltration, and endothelialization.

Results
Angiography of the grafts after one month showed at least partial patency in all grafts with the exception of one RGD-coated and one uncoated graft. Immunofluorescent staining of cell-sodded grafts showed a complete endothelial layer of adherent and spread PPAECs. Thrombus formation was present in two uncoated grafts and one RGD-coated graft. Intimal hyperplasia comprised mostly of smooth muscle cells was present in all grafts, but developed to the highest extent in RGD-coated grafts. Mean graft occlusion in M7 (24.7%) and cRRE-coated grafts (25.1%) was reduced by half compared to uncoated (58.5%) and RGD-coated grafts (53.9%).

Conclusions
Increased intimal hyperplasia in RGD grafts suggests that RGD, a prominent cell adhesion peptide, may facilitate ingrowth of more smooth muscle cells compared to the other grafts. These results show that cRRE-FSP coating of ePTFE grafts may reduce thrombosis, making it a promising candidate for improving graft patency.
Abstract 176: The Edge Effect: How Syndecan-1 Can Increase the Therapeutic Efficacy of Vasoregulatory Drugs and Growth Factors
Authors: Chaterji Somali , Gaddam Neha G , Atyam Neha , Ho Derek S , Baker Aaron B 
Published: March 2022

Rationale
Vascular smooth muscle cells (vSMCs) in mature animals normally reside in a quiescent state, expressing a distinct combination of contractile proteins, ion channels, and signaling molecules. However, in cases of vascular disease and injury, vSMCs are capable of being activated, shifting their phenotype through a continuum of cell fates. While the heparan sulfate proteoglycans syndecans, residing at cellular edges as transmembrane receptors, have emerged as powerful and complex regulators of cell function, their role in intercepting growth factors and therapeutic molecules modulating vSMC phenotype is unknown.

Objective
The purpose of this study was to examine the role of syndecan-1 in modulating the vSMC phenotype in the context of endogenous factors and drug treatments used for vascular disease.

Methods and Results
vSMCs were isolated from wild type (WT) and syndecan-1 knockout (S1KO) mice and cultured in different biochemical environments. Gene and protein expression studies indicated decreased α-smooth muscle actin (α-SMA) and calponin levels in S1KO cells relative to WT under simulated inflammatory conditions. Rapamycin treatment reversed the reduction in expression caused by S1KO, suggesting that the effects were, in part, mediated through mTORC1 signaling. In addition, S1KO vSMCs expressed higher levels of tissue factor, IL-6, and VCAM-1. These differences were maintained under treatment with heparin and TGF-β1, PDGF-BB, or atorvastatin. In-vivo studies with aortae from aged WT and S1KO mice indicated that S1KO arteries stained significantly lower for calponin.

Conclusions
Our findings not only provide insights into the fundamental cell surface receptors responsible for vSMC differentiation but also identify that the loss of syndecan-1 can make vSMCs more vulnerable to inflammatory molecular signals and potentially cause resistance to treatment with rapamycin and atorvastatin. Thus, preservation or enhancement of syndecan-1 expression may be a potential therapeutic target for altering the course of in-stent restenosis and atherosclerosis.
Abstract 178: Platelet Cd40 Exacerbates Atherosclerosis by Transcellular Activation of Endothelial Cells and Leukocytes
Authors: Lievens Dirk , Seijkens Tom , Kuijpers Marijke J , Winkels Holger , Hartwig Helene , Beckers Linda , Megens Remco T , Boon Louis , Noelle Randolph J , Gerdes Norbert , Heemskerk Johan W , Soehnlein Oliver , Weber Christian , Lutgens Esther 
Published: March 2022

Beyond their eminent role in hemostasis and thrombosis, platelets are recognized as mediators of inflammation and protagonists of atherogenesis. Here we investigated the inflammatory propensity of platelet-specific expression of CD40, the most common receptor for CD40L. Besides its presence on immune cells and other cell types, CD40 is constitutively expressed on platelets where its function is largely unknown.
CD40-deficient platelets exhibited impaired thrombin-induced activation, reflected by decreased P-selectin expression and αIIbβ3 integrin activation. Compared to infusion of activated Apoe-/- platelets, injection of activated Cd40-/-Apoe-/- platelets caused a >2-fold decrease in plaque size in the aortic arch (Apoe-/- platelets 18.3x104 ± 2.7x104 μm2 vs Cd40-/-Apoe-/- platelets 6.7 x104 ± 2.1 x104 μm2, Vehicle 9.8 x104 ± 2.8 x104 μm2, p<0.05) of Apoe-/- mice. Intravital microscopy of carotid arteries revealed impaired platelet adhesion and impeded leukocyte recruitment to the arterial wall of mice injected with activated Cd40-/-Apoe-/- platelets, which was accompanied by a decrease in endothelial VCAM-1 expression. Moreover, Cd40-/-Apoe-/- platelets displayed a reduced capacity to form platelet-leukocyte aggregates in vitro with these co-incubations releasing less IL-1β. Interestingly, once a thrombus had formed, leukocyte recruitment into thrombi was hampered in Cd40-/-Apoe-/- thrombi.
This study ascribes an essential function to platelet CD40 in atherosclerosis. Besides platelet activation, platelet CD40 induces activation of leukocytes and endothelial cells, thereby promoting inflammation and consequently atherogenesis.
Abstract 200: Geographical Location and Gender are Predictors of Elevated High Sensitive C-Reactive Protein (hsCRP) Levels and the Levels Strongly Correlate with Metabolic Syndrome (MetS) in a South Asian Population
Authors: Thanikachalam Mohan , Harivanzan V , Baliga Ragavendra R , Abraham William T , Thanikachalam Sadagopan 
Published: March 2022

Background
hsCRP is a risk factor for MetS. However, the distribution of hsCRP levels and its association with MetS in South Asians is unclear.

Methods
We conducted a population-based cross-sectional survey in 8,042 Indians (44% males, 56% females), age ≥20 yrs in urban (n=5, 017) and rural (n=3,025) communities. The study included anthropometric measurements, and fasting blood for plasma glucose, hsCRP and lipids. MetS was defined using NCEP (ATP III) criteria for Asians.

Results
The median hsCRP level was 2.30 mg/l [95% confidence interval (CI) 2.24, 2.39]. Adjusting for age and BMI, the hsCRP level was significantly higher in women, median 2.86 [CI 2.73, 2.97] when compared to men, median 1.80 [CI 1.71, 1.88] (p-value < 10-9). Adjusting for gender, age & BMI the urban communities had significantly higher hsCRP levels versus rural (p-value = 0.03). The fig. shows the distribution of hsCRP according to the number of components of MetS. The median hsCRP for those with 0, 1, 2, 3, 4, and 5 features of MetS are 1.01, 1.47, 2.3, 3.19, 3.70 and 3.94, respectively. Based on Kruskal-Wallis H test, after adjusting the hsCRP for gender, age and community, hsCRP levels were significantly higher in individuals with more components of MetS (p-value < 10-9 for trend). hsCRP and BMI were relatively independent predictors of MetS. Prevalence of MetS was higher in women (prevalence ratio [PR] = 1.15; CI 1.09, 1.21) and in urban communities (PR = 1.47; CI 1.35, 1.66).

Conclusions
hsCRP is highly associated with the MetS and the number of positive MetS components. Geographical location and gender are predictors of hsCRP levels, which is associated with a higher incidence of MetS in urban communities and women.