bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 132: Simulated Intraplaque Hemorrhage Stimulates Plaque Progression in an Animal Model
Authors: Chiu Stephanie E , Zhan James Q , Moody Alan R 
Published: March 2022

Intraplaque hemorrhage (IPH) is a feature of advanced plaques and a risk factor for clinical events. Several studies have demonstrated a link between carotid IPH and subsequent cerebrovascular events. Deposited RBCs contribute ingredients likely to promote plaque instability.
In this study, a catheter-based approach was used for intramural injection of RBCs into atherosclerotic plaques. We explored the hypothesis that an animal model of atherosclerosis demonstrates increased macrophage infiltration and neovascularization in plaques injected with RBCs.
Rabbits (n=4) were administered high cholesterol diet starting 2 weeks prior to endothelial denudation. Five weeks after denudation, a microinfusion catheter with a balloon-actuated microneedle delivered 70-100 μl of RBC and iodinated contrast mixture at multiple sites along the aorta. X-ray fluoroscopy and CT identified plaque injection sites and aortas were harvested for histopathology 5 weeks later.
Only sections from plaque injection sites (n=14) were positive for Perl’s iron stain. In the 3 aortas analyzed with immunohistochemistry (1 excluded due to plaque variability), plaque macrophage area proportion was significantly higher in plaque injection sites vs. non-injected vessel sections. Increase in plaque neovessel density was not significant. In conclusion, intraplaque deposition of RBCs in the atherosclerotic rabbit aorta increases measures of plaque instability.
Micrographs show Perl’s iron (a), macrophage (b), and neovessel (c) staining in an RBC-injected plaque. Scale bars represent 250 μm. Mean macrophage proportion and microvessel density and associated SEM are shown in bar graphs.
Abstract 137: Reversal of Endothelial Dysfunction Using Polyphenol Rich Foods and Supplements Coupled with Avoidance of Major Dietary Lectins
Authors: Gundry Steven R , Epstein Jean 
Published: March 2022

Endothelial dysfunction (ED) is highly correlated with the development of coronary artery and vascular disease in general. While dietary factors have been previously implicated in the causation of endothelial dysfunction, a purposeful change in dietary habits, coupled with the addition of several polyphenol-containing supplements, has not been studied for their effects on endothelial function, as measured by Peripheral Arterial Tonometry (PAT).
200 consecutive pts, aged 51-86, M:F ratio 3/2, with known vascular risk factors of HTN, DM, Hypercholesterolemia, hx of MI, Stent, CABG, were enrolled in a dietary program, which emphasizes large amts of leafy green vegetables, olive oil, radical reduction of grain, legumes, nightshades, and fruits; and generous amts of grassfed animal proteins, emphasizing Shellfish and avoiding commercial poultry (Diet Evolution). All pts were instructed to take 2-4,000 mg of high DHA fish oil, 200mg of Grape Seed Extract, and 50 mg of Pycnogenol per day. All pts had Endothelial Reactivity (ER) using PAT before and after a 5-minute arm occlusion using the EndoPAT 2000 (Itamar, Israel) at baseline and at 6 months.
Baseline Endothelial Reactivity was 1.88+/-0.7 (range 1.0-3.3), with 145/200 pts (72%)having endothelial dysfunction (less than 1.60). At 6 months, ER increased to 2.25+/-0.5 (range 1.2-3.6) (p<0.01). Only 40/200 (20%) remained with ED, but all had increased ER numbers. Ten pts stopped the polyphenols after a normal PAT; all developed ED on repeat PAT.
We conclude that a limited grain, legume, nightshade, and fruit diet with emphasis on greens, olive oil and grassfed animal proteins, particularly Shellfish, with avoidance of commercial poultry, with supplemental fish oil and polyphenols dramatically improves endothelial function in a group of high risk patients.
Abstract 146: Endothelin-1 Inoculation Attenuates Aortic Atherosclerosis in Rabbits under Cholesterol Rich Diet
Authors: Pineda Mauricio , Pineda Andres M , Maldonado Luz A , Uribe Ana M 
Published: March 2022

Introduction
The atherogenic potential of Endothelin-1 (ET1) has been extensively recognized and studied as a possible therapeutic target. However, the effects of an immunologic blockade of ET1 are unknown.

Hypothesis
Our hypothesis was that the induction of an immunological response through subcutaneous inoculation of Endothelin-1 prevents atherosclerosis in rabbits exposed to a cholesterol rich diet (CRD) for 36 weeks.

Methods
Eighteen New Zealand rabbits were divided in 4 different groups. Group A: 4 rabbits received CRD only. Group B: 5 rabbits were pre-treated with subcutaneous inoculation of ET1 (20 mg) plus Freund’s adjuvant (administered in 3 doses, 15 days apart) and then received CRD. Group C: 5 rabbits were pre-treated with subcutaneous inoculation of ET1 (20 mg) plus Montanide ISA 16 adjuvant (same schedule) and then received CRD; and group D: 4 rabbits received normal diet and no ET1 injection (control group). After 16 weeks, one rabbit in each group (A, B and C) and 2 rabbits in the control group were sacrificed. The rest of the animals were sacrificed after completing 36 weeks. An aortic extraction was done and pathologic slices prepared for hystomorphologic measurements, including vessel, lumen, and plaque area. Plaque/vessel area, plaque/lumen area, and plaque/lumen perimeter ratios, were also calculated.

Results
Compared with rabbits that received CRD alone, rabbits that were pre-treated with subcutaneous inoculation of ET1 plus Freund’s adjuvant had an statistically significant difference in the plaque area (0.9 ± 0.9 vs. 0.2 ± 0.3 mm2, p=0.03), plaque perimeter (2.4 ± 2.0 vs. 0.8 ± 1.1 mm2, p=0.04), plaque width (0.4 ± 0.3 vs. 0.2 ± 0.2 mm2, p=0.02), and calculated ratios (p<0.04) at 16 weeks. The difference was more significant after 36 weeks, with plaque area (1.3 ± 1.1 vs. 0.27 ± 0.5 mm2, p<0.001), plaque perimeter (4.6 ± 2.4 vs. 1.4 ± 1.5 mm2, p<0.001), plaque width (0.4 ± 0.2 vs. 0.2 ± 0.1 mm2, p<0.001), and calculated ratios (p<0.014). This gruop of rabbits also had the highest anti-ET1 antibody titers (1/12800).

Conclusions
Our findings suggest that the pre-treatment with subcutaneous inoculation of Endothelin-1 significantly attenuates aortic atherosclerosis in New Zealand rabbits exposed to a cholesterol rich diet for 16 to 36 weeks.
Abstract 150: Carotid Artery Atherosclerosis: Biomarkers of Plaque Instability
Authors: Simon Bartholomew V , Roztocil Elisa , Kumar Neil G , Caliste Xzabia , Doyle Adam J , Bandyopadhyay Gautam , Miller-Graziano Carol , Gillespie David L , Cullen John P 
Published: March 2022

Objective
To identify intraplaque and plasma biomarkers of carotid atherosclerotic plaque instability. We hypothesized that comparison of plaques and plasma from symptomatic and asymptomatic patients would show differences in proteolytic and inflammatory proteins, identifying them as potential markers of an unstable plaque.

Methods
42 symptomatic and asymptomatic patients (22 women, 20 men, 52-89 years) presenting for carotid endarterectomy had blood samples drawn and carotid plaques processed for protein extraction. The intraplaque expression of proteolytic enzymes (uPA, tPA, PAI-1 and MMP-2) and inflammatory markers (TF, TSP-1, and TNF-α) were evaluated by Western blot analysis. Plasma analyte concentrations were assayed with LUMINEX CVD1 and Cytokine panels using LUMINEX technology.

Results
Intraplaque expression of tPA and uPA was 5.9x and 4.5x higher (n=15, p< .05) respectively in the symptomatic group when compared to the asymptomatic group. The symptomatic group also demonstrated a 4.3x increase in active MMP-2 and a 3.6x increase in PAI-1 expression. Investigation of inflammatory proteins demonstrated 2.7-, 4.7-, and 3.3-fold increases in the expression of TNF- α, TSP-1, and TF, respectively. Plasma levels of MPO, MMP-9, IL8, MCP-1 and TNFα were significantly elevated in both the symptomatic and asymptomatic groups when compared to healthy controls (n=7, p<0.05). While there was no change in ICAM-1 levels between asymptomatic patients and healthy controls, symptomatic patients demonstrated a 35% decrease (n=7, p<0.05) when compared to controls. Also, plasma PAI-1 levels in asymptomatic patients were 55% lower than in symptomatic patients and 75% lower than in healthy controls.

Conclusion
Elevated expression of certain inflammatory markers and proteolytic enzymes in carotid plaques was demonstrated in symptomatic patients. Marked differences were also observed in ICAM-1 and PAI-1 expression in the plasma of symptomatic patients relative to asymptomatic patients and healthy controls. This leaves promising avenues for further inquiry into the roles of these proteins in atherosclerotic disease and their potential as clinical biomarkers of an unstable atherosclerotic plaque.
Abstract 161: Alternate Day Fasting With a High Fat and Low Fat Diet: Is Brachial Artery Flow Mediated Dilation Mediated by Adipokine Physiology?
Authors: Klempel Monica C , Kroeger Cynthia M , Phillips Shane A , Varady Krista A 
Published: March 2022

Background
Calorie restriction (CR) with variable macronutrient distributions has yielded inconsistent effects on brachial artery flow mediated dilation (FMD) related to weight loss and adipocyte physiology. Alternate day fasting (ADF) is a modified form of CR that has already elicited favorable changes in body weight and adipokine levels in both high fat (ADF-HF) and low fat (ADF-LF) background diets. However, the degree to which adipokines mediate vascular biology mechanisms on an ADF regimen is still unclear. This study examined if beneficial changes in body weight and adipokines influences endothelial function in ADF-HF and ADF-LF diets.

Methods
Thirty-two obese subjects were randomized to an ADF-HF (45% fat) or ADF-LF diet (25% fat), which consisted of two phases: 1) a 2-week baseline weight maintenance period, and 2) an 8-week ADF weight loss period. All food was provided to subjects.

Results
Body weight was reduced (P < 0.0001) by ADF-HF (5 ± 1%) and by ADF-LF (4 ± 1%). Fat mass decreased (P < 0.0001) by ADF-HF (5 ± 1 kg) and ADF-LF (4 ± 1 kg). Fat free mass remained unchanged. Waist circumference decreased (P < 0.001) by ADF-HF (7 ± 1 cm) and ADF-LF (7 ± 1 cm). FMD decreased (P < 0.05) in ADF-HF (2 ± 2%) and increased (P < 0.05) in ADF-LF (2 ± 2%). Adiponectin increased (P < 0.05) in ADF-HF (43 ± 7%) and in ADF-LF (51 ± 7%). Leptin decreased (P < 0.05) in ADF-HF (32 ± 5%) and in ADF-LF (30 ± 3%). Resistin decreased (P < 0.05) in ADF-HF (23 ± 5%) and ADF-LF (27 ± 4%). Increases in adiponectin were associated with augmented FMD post-treatment in the ADF-LF group only (r = 0.34, P = 0.03). Leptin and resistin were not correlated with changes in FMD.

Conclusion
Thus, an ADF-LF diet produces more favorable increases in FMD which are correlated with increased adiponectin as compared to the ADF-HF diet. These findings are important in terms of developing future behavioral modifications in energy restriction and macronutrient distribution that will improve vascular health.
Abstract 164: Action of Nicotinic Acid on Adiponectin, Leptin and Plasminogen Activator Inhibitor 1 Production and Expression In 3T3-L1 Adipocytes Under Oxygen Deprivation.
Authors: Nakamichi Renata , Quinto Beata M , Zanella Maria T , Batista Marcelo C 
Published: March 2022

Obesity has been considered one of the major contributors to the current global mortality. Recent studies suggest the dysregulation production of adipokines is partly responsible for obesity linked metabolic disorders.
The aim of this study was to investigate the effect of nicotinic acid (NA) on adipokines production and expression in 3T3-L1 adipocytes before and after hypoxia.
The mature adipocytes were pretreated with NA for 24h and induced by hypoxia for 4, 8 and 12h. Levels of adipokines in medium and total hypoxia inducible factor-1 alpha (HIF-1α) were quantified using immunoassay. Adipokines expression was analyzed by real time PCR.
Adipocytes treated with NA had lower leptin production in comparison to spontaneous production and an increase on leptin and Plasminogen Activator Inhibitor 1 (PAI-1) production associated with different periods of oxygen deprivation. It was observed a reduction of leptin levels in cells pretreated with NA compared to cells submitted to hypoxia. The same effect could be observed on PAI-1 production.
NA increased the production of adiponectin comparing to spontaneous production and cells pretreated witch NA increased adiponectin production during hypoxia.
Adipocytes were submitted to periods of oxygen deprivation had raised HIF-1α production comparing to spontaneous production and NA treatment. It also observed a reduction of HIF-1α in cells pretreated with NA and submitted to hypoxia.
Real time PCR showed a raise of adiponectin mRNA and a decreased PAI-1 in adipocytes treated with NA in comparison to spontaneous expression, but didn’t exert effect on spontaneous leptin.
Hypoxia, in all periods increased the leptin and PAI-1 expression and decreased adiponectin expression. In adipocytes pre-treated with NA blunted the production of leptin in spontaneous conditions and under hypoxia. It also increased PAI-1 production and expression. Adiponectin production and expression raised under NA treatment. The translation of such“in vitro” nicotinic acid benefits to the clinical setting should be object of future studies.
Abstract 185: Infliximab Suppresses TNF-a Induced Inflammatory Phenotype in Cerebral Vascular Smooth Muscle Cells: Implications for Cerebral Aneurysm Formation
Authors: Ali Muhammad S , Starke Robert M , Jabbour Pascal M , Tjoumakaris Stavropoula I , Gonzalez L. Fernando , Rosenwasser Robert H , Koch Walter J , Dumont Aaron S 
Published: March 2022

Background
Cerebral Aneurysm rupture results in significant morbidity and mortality. TNFα has been associated with cerebral aneurysms, but potential mechanisms have not been established, and its role in phenotypic modulation is unknown. Therapy targeting TNFα may represent a novel treatment strategy for cerebral aneurysms.

Hypothesis
Infliximab inhibits TNFα induced phenotypic modulation and upregulation of pro-inflammatory mediators.

Methods
Cultured cerebral VSMC were incubated with increasing concentrations of TNFα and gene expression measured with qPCR. Cerebral aneurysm induction surgery was performed in rats. Animal were euthanized at 4 wks for qPCR and immunohistochemical analysis of the circle of Willis. Animals undergoing aneurysm induction and Infliximab (5mg/kg) treatment were compared to animals undergoing aneurysm induction alone and untreated controls.

Results
In vitro experiments demonstrated suppression of differentiation markers and induction of pro-inflammatory genes. In vivo, TNFα mRNA and protein expression was significantly elevated following aneurysm induction compared to untreated controls. Expression of inflammatory markers including chemokines (MCP1 and VCAM1), transcription factors (NF-κB, KLF4), matrix remodeling proteins (MMP-2 & 3), IL-1β and iNOS was also increased. Furthermore, aneurysm induction resulted in phenotypic modulation by decreasing expression of SMC differentiation marker gene SM-α−Actin. Treatment with Infliximab significantly suppressed TNFα expression, inhibited expression of inflammatory markers, and decreased the number of CD68+ cells during aneurysm formation. Infliximab also reversed phenotypic modulation in vascular SMC following aneurysm induction.

Conclusions
TNFα plays a pivotal role in SMC phenotypic modulation with induction of pro-inflammatory genes and inhibition of SMC differentiation marker genes both in cultured cerebral vascular SMC and in vivo following cerebral aneurysm induction. Infliximab promotes anti-inflammatory effects in cerebral vessels by inhibiting TNFα and suppressing a pro-inflammatory cascade, which has important implications for the mechanisms behind intracranial aneurysm formation and potential therapeutic options.
Abstract 196: Effects of an Engineered Herpes Simplex Virus (HSV-1) on Vascular Cell Adhesion
Authors: McCormick Susan , He Qi , Weichselbaum Ralph , Skelly Christopher L 
Published: March 2022

Introduction
The genetically engineered HSV R7020 is known to block vascular smooth muscle cell (SMC) proliferation while allowing for re-endothelialization after balloon angioplasty. This HSV-1 anti-proliferative effect requires MEK activation. However, it is unknown what effect R7020 has on vascular inflammation and monocyte adhesion. We hypothesized that exposure of SMCs to HSV R7020 would not affect vascular adhesion molecules.

Methods
Human aortic SMCs were exposed to R7020 for 10 minutes in the absence and presence of TNF-α (10 ng/ml). After washing cells were cultured for 24 hours in the absence and presence of TNF-α. Vascular cell adhesion molecule 1 (VCAM-1) and intracellular adhesion molecue-1 (ICAM-1) levels were determined using immunocytochemistry and image J. The adhesion of calcein AM labeled PBMCs to these cells were also analyzed at 24 hours. VCAM-1 and ICAM-1 levels were measured in the carotids of New Zealand White Rabbits that underwent balloon injury and exposed to R7020 (n=4) or saline (n=4) for 10 minutes using microarray analysis.

Results

There was no significant difference in ICAM-1 levels in the R7020 treated and control SMCs (219±11 vs 223±1; p=0.758); there was an 8 fold increase in ICAM-1 in the TNF-α treated SMCs (219±11 vs 1587±68; p=1.2x10
-13
). Results were similar for VCAM-1. R7020 caused an increase in the adhesion strength of SMCs to collagen coated glass slides at 24 hours. Microscopic experiments (n=3) showed PBMCs adhered similarly to SMCs treated with TNF-α in the absence and presence of the virus (4 times that of control cells as measured by PBMC adhesion assay). Interestingly, after adhesion, Calcein AM was frequently observed in virus infected SMCs in the presence and absence of TNF-α. In cells not exposed to R7020, this rarely occurred. Microarray analysis demonstrated a 2.3 fold reduction in VCAM-1 expression in the R7020 treated rabbits (p=2.8x10
-14
) and no significant difference in expression in ICAM gene expression (p=0.3).

Conclusions
In contrast to TNF-α, R7020 does not increase ICAM-1 or VCAM-1. Both treatments domonstrate PBMC SMC adhesion with no additive or synergistic effect. However, R7020 results in intracellular transfer from PBMCs to SMCs suggesting the virus changes PBMC-SMC interplay.
Abstract 203: Smooth Muscle Cells from Abdominal Aortic Aneurysms are Unique and Can Independently and Synergistically Degrade Insoluble Elastin
Authors: Airhart Nathan , Brownstein Bernard , Cobb Perren , Schierding William , Arif Batool , Grapperhaus Kathy , Ennis Terri , Thompson Robert , Curci John A 
Published: March 2022

Background
The purpose of this study was to further elucidate the role of the vascular smooth muscle cell (SMC) in abdominal aortic aneurysm (AAA) disease. We hypothesized that that AAA-SMC are unique and actively participate in the process of degrading the aortic matrix.

Methods and Results
Whole-genome expression profiles of SMC from AAA, normal abdominal aorta (NAA) and carotid endarterectomy (CEA) were compared. We quantified elastolytic activity by culturing SMC in [3H]elastin-coated plates and measuring solubilized tritium in the media after 7 days. MMP-2 and MMP-9 production was assessed using real-time PCR, zymography and western blotting. Each SMC type exhibited a unique gene expression pattern. AAA-SMC had greater elastolytic activity than NAA (+68%, p<0.001) and CEA-SMC (+45%, p<0.001). Zymography showed an increase of active-MMP-2 (62kD) in media from AAA-SMC. AAA-SMC demonstrated 2-fold greater expression of MMP-2 mRNA (p<0.05) and 7.3-fold greater MMP-9 expression (p<0.01) than NAA-SMC. Culture with U937 monocytes caused a synergistic increase of elastolysis by AAA-SMC (41%, p<0.001) but not NAA or CEA (p=0.99). Co-culture with U937 caused a large increase in MMP-9 mRNA in AAA and NAA-SMC (p<0.001). MMP-2 mRNA expression was not affected. Western blots of culture media showed a 4-fold increase of MMP-9 (92kD) protein in AAA-SMC/U937 compared to NAA-SMC/U937 (p<0.001) and a large increase in active-MMP2 (62kD) which was less apparent in NAA/U937 media (p<0.01).

Conclusions
AAA-SMC have a unique gene expression profile and a pro-elastolytic phenotype that is augmented by macrophages. This may occur via a failure of post-transcriptional control of MMP-9 synthesis.
Abstract 217: Coronary Endothelial Dysfunction is Associated With Increased Risk of Osteoporosis
Authors: Prasad Megha , Reriani Martin , Khosla Sundeep , Lennon Ryan , Gulati Rajiv , Prasad Abhiram , Lerman Lilach O , Lerman Amir 
Published: March 2022

Background
A growing body of evidence links coronary artery atherosclerosis and calcification to osteoporosis in women. Coronary endothelial dysfunction (CED) is associated with adverse cardiovascular events. We aimed to study the association of CED and development of osteoporosis.

Methods
Coronary vascular reactivity was evaluated in 225 women, greater than age 50, with non-obstructive coronary arteries, by administration of intracoronary acetylcholine during diagnostic angiography. CED was defined as <50% increase in coronary blood flow (CBF) from baseline in response to maximal dose. After a median follow-up of 7 years, patients were assessed by a questionnaire for development of osteoporosis.

Results
The average age of the cohort was 60 years. Of 225 women, 60% (135) had CED. 18.2% (41 of 225 women) developed osteoporosis during follow-up. Women with CED were twice more likely to develop osteoporosis vs. women without CED (23% vs. 11.1%, p=0.024). Women who developed osteoporosis compared to those who did not had a markedly lower response (% change) in CBF: median (IQR)-4.3 (-28.3, 51.7) vs. 39.9 (-8.2, 106.0), p=0.007. In univariate analysis, CED was significantly associated with development of osteoporosis (p=0.024, relative risk 2.07, 95% CI: 1.07, 4.00). Multivariate analysis indicated CED as an independent predictor of osteoporosis (p=0.04).

Discussion
Endothelial dysfunction may account for or share similar mechanisms with development of osteoporosis in cardiac patients.