bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 34: Myh11 R247C is a Modifier Allele for Aortic Disease
Authors: Kwartler Callie , Kuang Shao-Qing , Prakash Siddharth , Huang Jian , Kamm Kristine , Stull James , Sweeney Lee , Milewicz Dianna 
Published: March 2022

Thoracic aortic aneurysms and dissections (TAAD) affect more than 30,000 Americans annually. TAAD can be inherited due to mutations in a number of genes, including MYH11, which encodes the smooth muscle myosin heavy chain. In non-familial TAAD patients, hypertension is the most common risk factor, but genetic contributors have not been explored. Exome sequencing has identified non-synonymous variants of unknown significance in MYH11 in the population, and these variants are enriched in TAAD patients without a family history. The goal of this study was to determine if rare MYH11 variants contribute to TAAD.
A knock-in mouse model of one recurrent rare variant, Myh11 R247C, shows no structural or pathologic abnormalities of the aorta, but does have decreased aortic contractility. We sought to determine if the Myh11 R247C allele could modify aortic disease due to another gene mutation. The Myh11R247C allele was bred into Acta2-/- mice, which develop aortic aneurysms, and into Acta2+/- mice, which do not. In both cases, the R247C allele significantly increases aortic diameter and accelerates elastin fragmentation and proteoglycan deposition; however the aneurysms do not progress to aortic dissection.
Treatment of Myh11R247C/R247C mice with a combination of L-NAME and high salt diet induced a sustained increase in systolic blood pressure from 130mmHg to 180mmHg. Approximately 25% of Myh11R247C/R247C mice on treatment died spontaneously with evidence of aortic dissections at necropsy, but without aortic dilation. The surviving mice also show no aneurysm formation. Aortic pathology shows significant fragmentation and thinning of elastin fibers, medial thickening, and a unique pattern of proteoglycan deposition along the elastin layers in the hypertensive Myh11R247C/R247C mice.
These data indicate that the Myh11R247C allele in mice increases severity of Acta2-/- associated aortic aneurysms and induces aortic disease when added to a known risk factor for TAAD, hypertension. Therefore, patients with the Myh11R247C rare variant and a disease-causing mutation may have earlier disease onset and increased severity; co-occurrence of the Myh11R247C allele in individuals with other benign genetic variants or risk factors like hypertension may induce TAAD.
Abstract 344: A20 Increases Endothelial Nitric Oxide Synthase Expression and Activity to Sustain Endothelial Cell Function under Inflammatory Insults
Authors: Chien Victor , Kaczmarek Elzbieta , Daniel Soizic , Essayagh Sanah , Lee Andy , Choi Lynn Y , Choi Jean Y , Da Silva Cleide G , Ferran Christiane 
Published: March 2022

Nitric oxide generated by endothelial nitric oxide synthase (eNOS) acts at different levels to sustain proper endothelial cell (EC) function. Pro-inflammatory molecules associated with various pathological conditions negatively affect eNOS expression and activity, altering vascular homeostasis. Decreased eNOS levels and activity are the pathognomonic features of EC dysfunction and a prelude to atherosclerotic remodeling of the vasculature. We have demonstrated that A20 maintains EC homeostasis by inhibiting NF-kappa B activation and protecting from apoptosis. The aim of this study is to explore whether A20 impacts eNOS expression/activity.
A20 was overexpressed in human coronary artery EC (HCAEC) by recombinant adenovirus-mediated gene transfer. Effects of A20 on eNOS expression and phosphorylation were analyzed by quantitative PCR, human eNOS promoter-luciferase reporter assay, chromatin immunoprecipitation (ChIP) and Western blot. Our data demonstrate that over-expression of A20 in HCAEC significantly increased eNOS mRNA and protein levels by promoting eNOS transcription, as demonstrated by ChIP, and eNOS promoter analysis using Luciferase reporter assays. Importantly, A20-induced upregulation of eNOS expression was sustained in HCAEC treated with tumor necrosis factor that inhibits eNOS transcription and function. Moreover, A20 enhanced phosphorylation of eNOS (Ser-1177), a surrogate marker of its activity, by promoting the activation of its upstream kinase AKT.
Our data demonstrate that A20 upregulates and activates eNOS, even in EC exposed to inflammation. A20’s ability to promote and sustain eNOS activity could be critical to its cytoprotective effect in EC. Further, it supports our pursuit of A20-based therapies to maintain vascular homeostasis, preventing vascular diseases such as atherosclerosis, transplant arteriosclerosis and diabetic vasculopathy.
Abstract 348: Fibrin Targeted in vivo Imaging of Tissue Plasminogen Activator-induced Thrombolysis of Mouse Deep Vein Thrombosis
Authors: Stein Ashley F , Kessinger Chase W , Zalada Castro Henry , Erdem S S , McCarthy Jason R , Lin Charles P , Jaffer Farouc A 
Published: March 2022

Introduction

Tissue plasminogen activator (tPA) is increasingly used to treat deep vein thrombosis (DVT) to prevent the post-thrombotic syndrome. However, while aging venous thrombi are likely to become resistant to fibrinolysis, the age-dependent response to fibrinolysis has not been systematically assessed. Here we used confocal intravital microscopy (IVM) to dynamically study fibrinolysis of murine thigh DVT at high-resolution using thrombus angiography and a fibrin-specific sensor, FTP11-CyAm7 (FTP11). We hypothesized that fibrinolysis could be tracked and quantified
in vivo
using serial IVM and molecular imaging of fibrin.

Methods
C57Bl/6 mice underwent topical 7.5% ferric chloride injury to induce femoral DVT. At day 1 (n=6) or day 4 (n=5) post DVT, IVM was performed during tPA infusion. Mice were i.v. injected with a bolus of tPA, heparin and plasminogen, followed by a continuous 1 hour infusion of tPA+heparin. FTP11, injected 45 min prior to lysis, enabled molecular imaging of fibrin. DVT area and length were quantified using FITC-dextran venography before and after lysis. IVM image stacks were analyzed by NIH ImageJ. After sacrifice, fluorescence microscopy (FM) and Carstair’s staining were performed.

Results
Compared to day 4, day 1 DVT fibrinolysis showed a greater net decrease in thrombus area (32±11 vs. 6±4%, p=0.004) and FTP11 signal (54±17 vs. 11±8%, p=0.004) after 60 min. Of note, day 1 more lyseable DVT had significantly higher baseline FTP11 signal (target-to-background ratio 1.91±0.86 vs. 1.10±0.06, p=0.004), consistent with greater thrombus permeability at day 1 vs. day 4. The net lysis correlated well with both the baseline FTP11 thrombus signal (r=0.62, p=0.048) and change in FTP11 signal (r=0.79, p=0.006). IVM angiography revealed a trend of increased lysis in the distal thrombus edge, compared to the middle or proximal edge (day 1: 44±11% vs. 26±16% and 29±26%, p>0.05). FTP11 signal colocalized with fibrin-rich thrombi on FM and Carstair’s staining.

Conclusion
Chemically induced murine DVT lyse more effectively at day 1 than day 4, as tracked by serial IVM molecular imaging. The FTP11-CyAm7 fibrin signal differentiates DVT of different ages and informs the magnitude of triggered fibrinolysis.
Abstract 358: Association of Adiponectin-with Cardio-metabolic Traits
Authors: Khan Hassan , Kunutsor Setor , Steur Marinka , Haycock Philip , Gregson John 
Published: March 2022

Background and Objectives
Current evidence on the association of adiponectin with fatal or non-fatal cardiovascular (CVD) outcomes is inconsistent. In addition association between adiponectin and related metabolic traits has not been systematically evaluated.

Design and Methods
A systematic search was performed using MEDLINE and Web of Science databases. Effect estimates and standard errors of associations of adiponectin with metabolic traits including (adiposity, glucose, insulin and cholesterol) and cardiovascular outcomes (coronary heart disease and stroke) were extracted from relevant prospective studies. The effect estimates and standardized correlation coefficients were pooled together using random effects meta-analysis.

Results
A total of 18 studies with 32,116 participants and 7,433 cardiovascular events reported on associations for adiponectin. Moderately strong and inverse associations were observed for adiponectin and measures of adiposity, body mass index, r=-0.13 [95% CI -0.29 to -0.02], waist to hip ratio, (r=-0.25 [95% CI -0.31 to -0.21] and waist circumference, r=-0.23 [95% CI -0.29 to -0.16]. Similarly inverse associations were seen with glucose, r=-0.19 [95% CI -0.22 to -0.17], insulin, r=-0.27 [95% CI -0.38 to -0.15] and triglycerides r=-0.31[95% CI -0.36, -0.26] while a positive correlation was observed with HDL-c, r=0.35[95% CI 0.30-0.41]. A pooled adjusted RR[95% CI] of 0.99 (95% CI: 0.86 to 1.14) with CVD for individuals in the top versus bottom third of baseline levels of adiponectin was seen. There was evidence of moderate heterogeneity between the findings of contributing studies (I2 =66%) with little difference explained by several study-level characteristics. Similarly modest associations were observed for both coronary heart disease (CHD), RR[95% CI] 0.89 (0.74 to 1.08) and stroke RR[95% CI] 1.01 (0.83 to 1.22).

Conclusion
Results of this systematic analysis give further evidence to the role of adiponectin with metabolic traits. However observational evidence from prospective studies indicates modest association with CHD and stroke outcomes that needs further investigation.
Abstract 378: Global MicroRNA Evaluation of The Brains and Aortas of a Prenatal Stress Murine Model With Vulnerable Plaque Show Sexual Dimorphism
Authors: Jhamnani Sunny , Aghili Nima , Lassance-Soares Roberta , Sood Subeena , Devaney Joseph , Burnett Mary Susan , Epstein Stephen 
Published: March 2022

Introduction and Hypothesis
Stress is a major cardiovascular risk factor. Preliminary studies in our lab demonstrated that prenatal stress in a murine model is associated with accelerated atherosclerosis and a vulnerable plaque phenotype at 20 weeks of age. Here, we hypothesized that prenatal stress is associated with global microRNA (miRNA) changes, possibly gender specific, and we sought to determine them by seeking such changes in two different tissues (brain and aorta) in both sexes.

Methods
Pregnant mixed background female mice were divided into stressed (chronic cold stress for the last 17 days of gestation) and non-stressed groups. Prenatally stressed and non-prenatally stressed offspring had urine corticosteroid levels measured at six weeks to see the effects of stress. At eight weeks the offspring were sacrificed and global miRNA levels were measured using the Affymetrix 2.0 miRNA chip on the brains and aorta.

Results
Urine corticosteroid levels were decreased in both genders at six weeks (p≤0.01), which has been shown in previous severe stress murine models. These were more significant in males (p=0.006) than in females (p=0.01). In eight-week-old males, there were 84 miRNAs in the brain that were dysregulated between prenatally stressed and non-stressed (p<0.05). The top miRNAs in males by p-value were miR-26, miR-21, and miR-346. Females that were eight weeks old had 89 dysregulated miRNAs (p<0.005) in the brain with miR-154 and miR-331 as the top miRNAs by p-value. Analysis of the aortas revealed 74 and 92 dysregulated miRNAs (p<0.05) with miR-326 and miR-15 as the top hits in males and females by p-value, respectively. Of interest, these top miRNAs have, in prior published studies, been implicated in playing a role in ischemia, ischemic preconditioning, stress and endothelial degeneration.

Conclusions
Prenatal stress leads to variety of miRNA changes that persist into at least the eighth postnatal week of life. These alterations in miRNA levels may contribute to the development of a vulnerable plaque phenotype that we have shown to be present at 20 weeks of age in a previous study. Prenatal stress also appears to exert gender specific effects, a finding that may provide insight into gender differences in cardiovascular outcomes.
Abstract 392: Effects of Dietary Omega-3 Polyunsaturated Fatty Acids Plus Minus Aspirin on Plasma Lipids and Hepatic Gene Expression in ApoE-Deficient Mice
Authors: Sorokin Alexander , Vaisman Boris , Thacker Seth , Remaley Alan 
Published: March 2022

BACKGROUND
Omega-3 fatty acids (O3FA) have hypolipemic, anti-inflammatory and atheroprotective effects. Aspirin (ASA) may potentiate these beneficial effects, by altering the production of anti-inflammatory mediators, such as resolvins.

OBJECTIVE
To compare the anti-atherogenic effects and gene expression changes of O3FA diet plus minus ASA.

METHODS

12-week old apoE-Ko females were placed for 2 weeks on a O3FA-deficient (O3FA-) diet, and were then treated for 13 weeks with either O3FA-, O3FA+ or O3FA+ASA diet (N=10). RNA from liver (N=4) was isolated and analyzed by Qiagen PCR Array for 84 genes related to cholesterol metabolism. Arteriosclerosis was assessed by
en face
analysis.

RESULTS

O3FA+ diet decreased plasma total cholesterol, triglycerides, phospholipids, free cholesterol and cholesteryl esters between 26%-39% (
P
<0.0005). Majority of the decrease in cholesterol was due to a lowering of VLDL-C and LDL-C. Similar lipoprotein changes were observed in mice on the O3FA+ASA diet. O3FA+ fed mice showed numerous significant changes in the expression of genes related to cholesterol metabolism. Insig1 was decreased by 1.9-fold, whereas Nr1h4 and Prkaa1 were increased by 1.5 fold. Cholesterol biosynthesis genes, such as Dhcr7, Fdps and Hmgcr were decreased in 1.6, 2, and 1.4-fold, respectively. Expression of Acaa2 and Hmgcs2, genes involved in catabolism of cholesterol, were increased by 1.7-1.8 fold. O3FA+ASA treated mice showed a similar hepatic gene expression profile except PCSK9, which decreased 5.9-fold in these mice but only by 2.5-fold in O3FA+ treated mice. Compared to O3FA- treated control mice, O3FA+ASA treated mice showed 2.2-fold decrease in aortic atherosclerosis, whereas no change compared to control mice was observed with just the O3FA+ diet.

CONCLUSIONS
Addition of ASA to the O3FA+ diet showed a greater effect in lowering the expression of PCSK9 in the liver and decreased aortic atherosclerosis. ASA treatment, however, did not show a significant further improvement in the lowering of plasma pro-atherogenic lipoproteins over that observed with the O3FA+ diet, suggesting that the anti-inflammatory property of ASA may account for its anti-atherogenic effect.
Abstract 465: Heparanase Alters Endothelial Phenotype, Leukocyte Attachment and Atherosclerotic Plaque Formation
Authors: Johnson Collin G , Bondada Vidya , Yeager Doug , Emelianov Stanislav , Baker Aaron 
Published: March 2022

Little is known about the role of enzymatic regulation of glycosaminoglycans in the formation of atherosclerotic plaques. We tested the hypothesis that heparanase (HPA) could regulate endothelial cell inflammatory responses and immune cell mobilization, promoting atherogenesis.

We transduced human endothelial cells with lentiviral vectors for one of four constructs expressing shRNA targeted to HPA or a scrambled control sequence. These shRNA knocked down HPA expression 30-50% of wild type levels. We also overexpressed HPA in these cells using lentiviral constructs. Finally, we transduced cells to express miRNA 1258 known to reduce levels of HPA in endothelial cells. These cell lines were confirmed for gene modulation using PCR and western blotting for HPA, then assayed for the expression of inflammatory markers such as ICAM, VCAM, and IL-8 under static and physiologic levels of shear stress (12 dynes/cm
2
). Increasing HPA levels led to a corresponding increase in inflammatory cytokines in endothelial cells, while reduction of HPA levels by targeted shRNA or miRNA caused reduced expression. Using fluorescently labeled monocytes we exposed the endothelial cell layer to circulating leukocytes in an in-vitro flow system. Adhesion of monocytes increased in cell lines with enhanced HPA expression and decreased in those with HPA knockdown.

We also evaluated the effects of low endothelial sheer stress in transgenic mice overexpressing the human HPA gene crossed with the ApoE knockout mouse line. We surgically implanted an extravascular cuff on the left carotid artery of mice from this HPA-Tg/ApoE line and the ApoE line to create a region of disturbed flow and low shear stress. We used high resolution combined ultrasound and photoacoustic imaging system to image the flow and plaque morphology after 1 day, 2 weeks and 4 weeks post-implantation. Doppler imaging revealed overexpression of HPA increases the formation and severity of atherosclerosis. The oscillatory flow in the carotid artery due to the cuff placement lead to increased plaque accumulation and increased inflammation in response to the induced shear stress in the HPA-Tg/ApoE mice compared to the ApoE control mice.
Our studies support HPA as a potent regulator of inflammation and plaque formation.
Abstract 475: Cationic Peptides in Inflammation and Atherosclerosis
Authors: Selvarajan Krithika , Aluganti Narasimhulu Chandrakala , Parthasarathy Sampath 
Published: March 2022

Background
Apoprotein (Apo A1) and apoprotein E mimetic peptides are known to reduce atherosclerosis. Antioxidant, anti-inflammatory, and hypolipidemic properties have been suggested to contribute to the anti-atherosclerotic effects. In this study, we tested whether three distinct and unrelated cationic peptides would inhibit inflammatory response to lipopolysaccharide (LPS) and oxidized low density lipoprotein (Ox-LDL).

Methods
5F-mimetic peptide of apoA1, LL27 derived from the anti-microbial peptide CAMP, and a human glycodelin derived peptide were commercially synthesized. RAW 264.7 cells were incubated with LPS or Ox-LDL, pretreated with and without peptides. RNA was isolated from treated cells and real time PCR was performed using mouse IL-1α and IL-6 primers.

Results
Increased IL-1α and IL-6 gene expression was observed in the presence of LPS/Ox-LDL. However, peptides showed dual effect as they induced inflammatory markers in the presence of LPS except for LL27 but all peptides inhibited the Ox-LDL-induced inflammatory gene expressions. The results suggest that these peptides may have dual effects on inflammatory cytokine genes in the presence of LPS and Ox-LDL.

Conclusion
Based on these results, we predict that lysine-rich cationic peptides could have therapeutic potential in reducing CVD/atherosclerosis-associated inflammation. However, specific receptors, presence of lipids, and other factors may influence their effects.
Abstract 82: Macrophage Specific ABCA1 Deletion Does Not Worsen Insulin Resistance in High Fat Diet Induced or Genetic Obese Mouse Models
Authors: Zhu Xuewei , Bi Xin , Chuang Chia-Chi , Chung Soonkyu , Brown Amanda L , Cuffe Helen , Gebre Abraham K , Boudyguina Elena , Parks John S 
Published: March 2022

Introduction
Obesity associated low-grade, chronic inflammation underlies the pathogenesis of type 2 diabetes and insulin resistance. ATP-binding cassette transporter A1 (ABCA1), a membrane lipid transporter, forms nascent HDL particles by effluxing cellular free cholesterol (FC) and phospholipids to lipid-poor apolipoprotein A-I. In macrophages, ABCA1 also dampens pro-inflammatory MyD88-dependent Toll-like receptor signaling by reducing cellular membrane FC and lipid raft content, indicating a role for ABCA1 in macrophage-mediated innate immunity. However, the role of ABCA1 in macrophage-associated innate immunity in insulin resistance is unclear.

Hypothesis
We tested the hypothesis that macrophage-specific ABCA1 deletion may exacerbate insulin resistance by increasing obesity-induced chronic, low-grade inflammation.

Methods and Results
Wild type (WT) and macrophage-specific ABCA1 knockout (MSKO) mice in the C57/BL6 background were fed a 45% High Fat Diet (HFD) for 16-24 weeks or crossed into the leptin deficient (ob/ob) background to induce obesity. Our results showed that MSKO and WT mice developed obesity, insulin resistance, hypercholesterolemia, and hepatic steatosis to a similar extent with HFD feeding or after crossed into the ob/ob background. Resident peritoneal macrophages from HFD-fed or leptin deficient MSKO vs. WT mice accumulated significantly more cholesterol. Relative to chow, HFD markedly induced macrophage infiltration and inflammatory cytokine expression to a similar extent in adipose tissues in WT and MSKO mice. Among the examined pro-inflammatory cytokines in peritoneal macrophages, only IL-6 was highly up-regulated in ob/ob MSKO vs. WT control mice, indicating that lack of ABCA1 in macrophages does not significantly affect obesity associated chronic inflammation.

Conclusion
Although macrophage specific ABCA1 deficiency enhances Toll like receptor signaling, it does not significantly exacerbate obesity associated low grade, chronic inflammation, therefore has no impact on the pathogenesis of insulin resistance in both HFD induced and genetic obese mouse models.
Abstract 99: The Effect of Androgen Deprivation Therapy on Conduit Artery Endothelial Function in Men with Prostate Cancer
Authors: Zuflacht Jonah , Milian Jessica , Kadivar Samoneh , Graham Powell , Nguyen Paul , Beckman Joshua A 
Published: March 2022

Introduction
Androgen deprivation therapy (ADT) is a standard treatment for patients with aggressive prostate cancer. While ADT improves cancer-specific and overall survival, it can increases insulin resistance and the risk of diabetes. Emerging evidence suggests that ADT also increases the incidence of adverse cardiovascular events in patients with cardiovascular disease, but the exact mechanism is unknown.

Hypothesis
We hypothesized that ADT may impair endothelial function due to increases in insulin resistance and provide a mechanism for heightened cardiovascular risk in this population.

Methods and Results
We prospectively evaluated conduit artery function in 10 consecutive men (mean age 65 ± 7 years; 30% with diabetes) with prostate cancer before and 3 months after initiation of ADT. High-resolution B-mode ultrasound was used to assess flow-mediated (endothelium-dependent) and nitroglycerine-mediated (endothelium-independent) brachial artery vasodilation. Endothelium-dependent vasodilation was greater at 3 months compared with baseline (11.9 ± 1.7% vs. 9.23 ± 1.8%, p = 0.002). There was no significant change in resting arterial diameter, reactive hyperemic stimulus, mean arterial pressure, nor heart rate between study conditions. Nitroglycerine-mediated, endothelium-independent vasodilation did not change from baseline (18.3 ± 3.55%) to the three month visit (19.7 ± 2.40%), p = 0.466). Markers of insulin resistance will be available at the time of the meeting.

Conclusions
In contrast to our expectation, ADT improves endothelial function. This suggests that the physiological mechanism linking ADT with increased cardiovascular events is not likely regulated by the vascular endothelium. Moreover, this mechanism of insulin resistance may be beneficial to endothelial function.