bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 596: Platelet inhibition by Ticagrelor in African American versus Caucasian patients after Percutaneous Coronary Intervention
Authors: Katikaneni Pavan K , Modi Kalgi
Published: July 2021

PLATO study showed that in acute coronary syndromes, treatment with Ticagrelor as compared with Clopidogrel significantly reduced the rate of death, myocardial infarction or stroke. In subgroup analysis based on race, no significant difference in primary end point was noted. However, African Americans (AA) comprised only 1.1% of study population. AA have greater mortality from coronary artery disease (CAD) and more likely to have high on-treatment platelet reactivity. Aim of our study is to compare the extent of platelet inhibition achieved by Ticagrelor and Clopidogrel in AA versus Caucasian patients undergoing percutaneous coronary intervention (PCI). Methods: 72 patients undergoing PCI during September to November 2014 were included in this study. All received Aspirin 325mg loading dose (LD) followed by 81mg maintenance dose, along with either Ticagrelor 180 mg LD followed by 90 mg twice-daily MD or Clopidogrel 600 mg LD followed by 75 mg once-daily MD. Platelet reactivity (PRU) was measured using the VerifyNow P2Y12 function assay 12-24h following MD and the average values calculated for each group. Results: 40 African American (55%) and 32 Caucasian (45%) patients were included. Ticagrelor was used in 45 (62.5%) and Clopidogrel was used in 27 patients (37.5%). Among Caucasians, 16 (50%) received Ticagrelor and 16 (50%) received Clopidogrel. Among AA, 27 patients (67%) received Ticagrelor, 13 patients (33%) received Clopidogrel. In the total study population, the average PRU achieved was lower for Ticagrelor (44) compared with Clopidogrel (183). Among patients who received Ticagrelor, significantly lower average PRU was seen in AA (33) compared to Caucasians (63). Among patients who received Clopidogrel, similar average PRU was seen in AA (181) and Caucasian (185) groups. After combining Ticagrelor and Clopidogrel groups, lower average PRU was achieved in AA (76) compared to Caucasians (126), the difference primarily attributable to Ticagrelor. Conclusion: Ticagrelor achieved greater platelet inhibition in AAs compared to Caucasians, while Clopidogrel achieved similar platelet inhibition in both racial groups. This potentially greater platelet inhibition by Ticagrelor in high risk AA group merits investigation in large-scale studies.
Abstract 597: Plasma Fibrin Clot Strength in Patients With Diabetes Mellitus Measured by Thrombelastography
Authors: Maatman Benjamin , Schmeisser Glen , Owens Janelle , Flockhart David , Kreutz Rolf P
Published: July 2021

Background:
Patients with diabetes mellitus (DM) exhibit increased risk of recurrent myocardial infarction. Maximal clot strength measured by thrombelastography (TEG) in whole blood is a risk factor for recurrent ischemic events. We hypothesized that diabetic subjects exhibit increased fibrin clot strength in platelet poor plasma and that extent of glycemic control correlates with maximal fibrin clot strength.

Methods:
Platelet poor plasma samples were obtained from subjects with known or suspected coronary artery disease prior to cardiac catheterization (n=354). Kaolin activated TEG was performed in citrate plasma. Time to fibrin formation (R), clot formation time (K), and maximal fibrin clot strength (MA) measured. Hemoglobin (Hgb) A1C and daily administered insulin doses were recorded.

Results:
Fibrin MA was increased among subjects with DM as compared to non-DM (37.0 ± 8 mm vs. 34.1 ± 8; p<0.001). HgbA1c was significantly correlated with MA (r=0.213; p=0.002). There was no significant correlation between daily insulin dose and MA among patients with insulin dependent DM (r=0.04; p=0.8). In multivariable regression analysis, DM remained significantly associated with plasma MA after adjustment for other clinical variables (p=0.01).

Conclusions:
DM is associated with elevated fibrin clot strength (MA). In addition, increased MA appears to be associated with worse glycemic control as defined by HgbA1C. Our results support that patients with DM, in particular those subjects with poor control, demonstrate more stable fibrin generation than subjects without DM. This may in part contribute to the increased ischemic risk in patients with poorly controlled DM.
Abstract 621: Homocysteine Activates T Cells by Enhancing Endoplasmic Reticulum-mitochondria Coupling and Increasing Mitochondrial Respiration
Authors: Feng Juan , Wang Xian
Published: July 2021

Hyperhomocysteinemia (HHcy) accelerates atherosclerosis by affecting the immuno-inflammatory response, increasing proliferation and stimulating cytokine secretion in T cells. However, whether homocysteine (Hcy) activation of T cells is associated with metabolic reprogramming is unclear. Here, we showed that Hcy (50 μM, 24 hr)-stimulated splenic T-cell proliferation in mice was accompanied by increased levels of mitochondrial reactive oxygen species (ROS) by 23.25±2.27%, calcium overload, increased mitochondrial mass by 24.29±7.97% and increased respiration. Inhibiting mitochondrial ROS levels and calcium signals or blocking mitochondrial respiration largely blunted Hcy-induced T-cell activation. Hcy also enhanced endoplasmic reticulum (ER) stress in T cells. Inhibiting ER stress with 4-phenylbutyric acid or mitochondrial respiration by rotenone blocked Hcy-induced T-cell proliferation and interferon-γ (IFN-γ) secretion. Mechanistically, Hcy treatment increased ER-mitochondria coupling as revealed by structured illumination microscopy and elevated expression of tethering proteins MFN2, Gpx7, and ERP44. Uncoupling ER and mitochondria by the microtubule inhibitor nocodazole attenuated Hcy-stimulated mitochondrial ROS production, calcium overload, mitochondrial membrane potential, IFN-γ secretion and T-cell proliferation; thus, juxtaposition of ER and mitochondria is required for Hcy-promoted mitochondrial function and T-cell activation. In conclusion, Hcy promotes T-cell proliferation and IFN-γ secretion by inducing metabolic reprogramming via regulating ER-mitochondrial coupling. Our results highlight the importance of metabolic regulation in T-cell activation and shed new light on understanding the pathogenesis of HHcy-accelerated atherosclerosis.
Abstract 622: Inflammation and Arterial Compliance: A Paradox? The Bogalusa Heart Study
Authors: Fernández Alonso Camilo , Stuchlik Patrick , Barshop Rupert , Galazka Patrycja , Blandon Roberto , Li Shengxu , Chen Wei , Berenson Gerald S
Published: July 2021

Introduction:
Systemic inflammation -as measured by high-sensitivity C-reactive protein (hs-CRP)- is adversely associated with arterial compliance. However, information is scant on whether this association in consistent throughout the different non-invasive measurements of arterial compliance.

Hypothesis:
We assessed the hypothesis that divergences exist in the association between hs-CRP and measurements of arterial compliance, and that this association differs among race (white/black) groups in relatively young adults.

Methods:
Measurements of hsCRP and non-invasive arterial compliance --large-artery elasticity index (C1), small-artery elasticity index (C2), brachial-ankle pulse-wave velocity (ba-PWV) and augmentation index (AI@75)-- were assessed in 702 non-institutionalized participants with a mean age of 43.5 years (29.4-51.1 years); 70.8% whites and 43.7% males, as a part of the Bogalusa Heart Study. Race-specific independent associations were tested through multivariable-adjusted linear regression analyses.

Results:
Black vs white participants had higher hs-CRP, baPWV and AI@75 (p<0.01); whereas C1 and C2 were higher in whites (p<0.01). In multivariable-adjusted linear regression analyses, controlling for: age, sex, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), body mass index (BMI), smoking status and other traditional cardiovascular risk factors, hs-CRP was significantly and independently associated with AI@75 (β=0.11, p<0.01) and C1 (β=-0.08, p=0.04), in whites only. In contrast, black participants did not show any significant associations among these parameters. C2 and baPWV did not exhibit association with either race group.

Conclusion:
In conclusion, these findings help enhance the concept that the association of inflammation and arterial compliance is seemingly relative, as it is dependent on the measurement used to assess the latter; with its impact varying by race (black-white). Further, these observations may aid in revising existing methodologies used in the diagnosis of inflammation-mediated structural and functional damage, in addition to enhance race-specific approaches for screening and prevention of cardio-metabolic risk factors.
Abstract 639: Carnosine Supplementation Promotes Revascularization in Mouse model of Hindlimb Ischemia
Authors: Boakye Adjoa A , Conklin Daniel , Guo Luping , Zhang Deqing , McCracken James , Bhatnagar Aruni , Baba Shahid P
Published: July 2021

Revascularization surgeries are important for treatment of critical limb ischemia (CLI) however, advanced CLI patients are poor candidates for these surgeries. Here, we proposed that carnosine, a dipeptide of β-alanine and histidine which is highly abundant in the skeletal muscle (2-20mM) and has the abilities to quench reactive aldehydes and chelate divalent metals, can promote revascularization. WTC57Bl/6 mice were pre-treated with carnosine (1g/L) in drinking water for 7 days, subjected to hindlimb ischemia surgery (HLI) by femoral artery ligation and allowed to recover for 14 days with carnosine supplementation. Doppler analysis showed that carnosine supplementation increased blood perfusion to 63±6% compared to 19±3% in controls. Post-surgery carnosine treatment also increased blood flow to 49.4± 6% compared to 28.2± 4% in controls following 21 days of recovery. Histological analysis of the ischemic limb showed that tissue injury was attenuated by carnosine supplementation. Mobilization of angiogenic endothelial progenitor cells (EPCs) (Flk+/Sca+) and the expression of vascular endothelial growth factor (VEGF) in the ischemic muscle was enhanced by carnosine treatment. These results were associated with an increase in ischemic muscle carnosine levels in the carnosine treated HLI mice. Furthermore, expression of the carnosine transporter PHT1 was increased in carnosine treated HLI mice compared to HLI mice. Concomitant with the increased VEGF expression in carnosine treated HLI mice, we observed that pre-treatment of murine myoblast C2C12 cells with octyl-D-carnosine followed by hypoxia increased hypoxia inducible factor 1-alpha nuclear translocation and VEGF expression and secretion. Our results demonstrate that carnosine treatment increases blood flow in the ischemic limb by increasing EPC mobilization and therefore can be a potential therapeutic intervention for CLI patients.
Abstract 645: Heparin’s Effects on Vascular Cells Require Transmembrane Receptor 184A
Authors: Farwell Sara Lynn N , Barthol Trista , Slee Joshua B , Lowe-Krentz Linda J
Published: July 2021

Introduction:
Though heparin has been used in the clinic for decades, molecular mechanism(s) underlying heparin’s functions independent of anti-coagulation are still unclear. Transmembrane protein 184A (TMEM184A) is conserved, yet its physiological and molecular functions remain unknown. There are few studies reporting its expression and potential role in membrane trafficking in exocrine cells, and involvement in signaling during male germ cell differentiation. Sequence analysis reveals that the C terminal domain includes a putative binding site for heparin. We have previously shown that heparin decreases proliferation in vascular smooth muscle cells (VSMCs) by inducing expression of MKP-1 that decreases Elk-1 and ERK activation.

Hypothesis:
We hypothesized that TMEM184A plays a role in mediating heparin effects in vascular cells.

Methods and Results:
We observed TMEM184A expression in vascular cells through immunofluorescence and western blotting using three primary antibodies against different regions in TMEM184A, and visualized cells with confocal microscopy. To investigate whether heparin effects were dependent on TMEM184A, VSMCs were electroporated with 20 μg/ml control or TMEM184A shRNA. Control and knockdown VSMCs were treated with 200 μg/ml heparin for 20 min followed by 2 μg/ml platelet-derived growth factor (PDGF). Activated ERK or Elk-1 in the nucleus was compared to untreated controls or cells treated with PDGF alone. Quantitative immunofluorescence of over 100 cells for each treatment from at least three independent experiments showed that heparin treatment prior to 20 min PDGF stimulation significantly decreased active ERK by nearly 50% in control shRNA cells compared to cells treated with PDGF alone (20 min PDGF = 100.0% ± 3.76%, heparin pre-treatment = 55.5% ± 2.20%; p<0.001). In TMEM184A knockdown cells, heparin pre-treatment did not decrease ERK activation (20 min PDGF = 100.0% ± 3.27%, heparin pre-treatment = 109.8% ± 3.06%). Similar results were observed for Elk-1. Heparin also did not decrease proliferation in response to PDGF in knockdown VSMCs as shown with BrdU incorporation assays.

Conclusions:
Our results provide functional evidence that heparin signaling in VSMCs is mediated at least in part by TMEM184A.
Abstract 656: Lipid Phosphate Phosphatase-3 Mediated Inflammation and Angiogenesis
Authors: Miriyala Sumitra , Maxey Benjamin , Duhon Megan , Steven Alexander Jonathan , Escalante-Alcalde Diana , Smyth Susan , Day Alicia , Fang Xianjun , Panchatcharam Manikandan
Published: July 2021

A recent meta-analysis of genome-wide association studies (GWAS) of coronary artery disease (CAD) involving over 86,000 individuals identified PPAP2B (gene that encodes lipid phosphate phosphatase-3; LPP3) as 1 of 13 new loci that associate with CAD. The PPAP2B loci independently predicted CAD (P = 3.81 X 10-19) and lacked association with traditional risk factors. The lipid phosphate phosphatase 3 is a cell surface integral membrane protein that regulates the bio availability of major bioactive lysophospholipids, like lysophosphatidic acid (LPA). Our group also made the surprising discovery that tissue-specific deficiency of LPP3 exaggerates vascular inflammation and promotes the development of neointima after vascular injury. Here we present evidence that enothelial cell LPP3 serves as an intrinsic negative regulator of vascular inflammation, promotes endothelial barrier function. Our results suggest that loss of LPP3 by siRNA mediated knockdown enhanced proliferation with elevated pErk signaling whereas migration was inhibited in human microvascular endothelial cells (HMEC). The endothelial cells that were silenced for LPP3 were further subjected LPA receptor knockdown and we found that although LPA-induced Erk phosphorylation was similar between LPA
1
and LPA
4
silencing, LPA
4
silencing resulted in enhancement of Akt phosphorylation at Thr-308 in response to LPA. Dual inhibition of LPA
1
and LPA
4
abolished LPA mediated migration in LPP3 silenced HMEC suggesting that the signaling response is mediated through the LPA
1
receptor. Rac activation in response to LPA was barely detectable in LPP3 silenced HMEC, which correlated with its weak migratory response. In contrast, a prominent increase in Rac-GTP levels was observed upon LPA
4
silencing under these conditions. Activation of HMEC with LPA increased phosphorylation of MSK downstream of p38, and CREB phosphorylation was inhibited upon LPA
4
receptor blockade in LPP3 silenced endothelial cells.
Abstract 658: Beta-Catenin C-terminal Signaling Suppresses p53 in Vascular Smooth Muscle Cells and is Essential for Arteriogenesis
Authors: Riascos Bernal Dario F , Chinnasamy Prameladevi , Valenta Tomas , Basler Konrad , Sibinga Nicholas E
Published: July 2021

Arteriogenesis requires smooth muscle cell (SMC) investment of endothelial tubes to build the arterial wall. In preliminary studies in mice, inactivation of SMC β-catenin, a downstream mediator of canonical Wnt signaling and a key element of the structural cadherin-catenin complex, caused embryonic demise with apparent vascular enlargement and hemorrhage. In detailed analysis, we found that SMC β-catenin is required for arteriogenesis: embryos lacking SMC β-catenin (SMKO) died by E12.5 (93 embryos and 215 pups analyzed) with impaired arterial maturation, with thinner aortic walls by E10.5 (SMKO vs. WT p<0.0001, n=8) due to impaired SMC investment, decreased cell proliferation (E9.5 to E11.5 SMKO vs. WT p5), and increased apoptosis (E11.5 SMKO n=11 vs. WT n=18, p<0.05). Interestingly, β-catenin-deficient SMCs had impaired growth and survival and increased p53 activity, though p53 levels were unchanged. In vivo, SMC p53 inactivation suppressed phenotypes caused by loss of β-catenin: at E12.5, no β-catenin SMKOs looked normal, with 21% and 79% in early and advanced resorption, respectively; in contrast, 23% of p53/β-catenin double SMKOs looked normal, with 36% and 41% in early and advanced resorption, respectively (β-catenin SMKO n=29 vs. double SMKO n=22, p<0.01). Double SMKOs showed marked recovery of arterial wall formation. Mechanistically, signals from β-catenin’s C-terminal, but not N-terminal, interaction domain repressed p53 transcriptional activity in cultured SMCs (p<0.01, n=3). In vivo, studies of three different β-catenin knockin mutants showed that β-catenin signaling was required in SMCs, while structural function was not sufficient for arteriogenesis, as a structurally-competent but signaling-inert mutant β-catenin (n=11) phenocopied the SMKO (n=29). Moreover, signals from the C-terminal interaction domain were both required and generally sufficient for arteriogenesis: expression of a C-terminus β-catenin mutant in SMCs caused embryonic lethality (83 pups analyzed), while an N-terminus mutant permitted normal development (101 pups analyzed). Thus, SMC β-catenin-mediated repression of p53 is required for arteriogenesis, and its C-terminal interaction domain is important for this novel developmental role.
Abstract 659: Inhibition of vWF Secretion with Novel G alpha12 N-terminal alpha-SNAP Binding Domain Peptide Increases Survival in Septic Rodents
Authors: Rusu Luiza , Schlapfer Martin , Offermanns Stephan , Du Xiaoping , Minshall Richard D
Published: July 2021

Severe sepsis is associated with disseminated intravascular coagulation (DIC) as a result of interdependent mechanisms of systemic intravascular inflammation, microvascular thrombosis, and thrombocytopenia. Currently, no drug is available to concomitantly treat these events in sepsis. A poorly understood mechanism and yet critical determinant of sepsis-induced microvascular thrombosis is von Willebrand Factor (vWF) secretion by activated endothelial cells. We recently discovered that heterotrimeric G protein alpha subunit G alpha 12 plays a critical role in basal and evoked vWF secretion by endothelial cells by promoting Weibel-Palade body (WPB) exocytosis. Based on the observed interaction of G alpha 12 with alpaSNAP, a critical member of the exocyst complex required for plasma membrane fusion and exocytosis of WPB contents, we generated a myristoylated Galpha12 N-terminal alphaSNAP Binding Domain (Myr-SBD) blocking peptide and tested the hypothesis that this would selectively and potently inhibit Galpha12 interaction with alphaSNAP and thereby block vWF secretion, limit platelet adhesion and prevent microvascular thrombosis associated with cecal ligation and puncture (CLP) induced sepsis. CLP-induced fulminant sepsis in rats and mice was associated with a 2-3-fold increase in plasma vWF within first 24 hrs. Importantly, we observed reduced plasma vWF levels 24 hrs after CLP surgery in mice given a one-time i.v. bolus (2 μmol/kg) of micellar Myr-SBD at the time of surgery as compared to Myr-scrambled peptide or vehicle only group. Strikingly, this was associated with increased survival without adversely inducing hemorrhage and vascular leakage. Furthermore, and consistent with the hypothesis that Gα12-dependent increase in vWF secretion during sepsis leads to poor outcome, control WT mice succumbed to sepsis in less than 96 hrs whereas 80% of Gα12-/- mice shown previously to have significantly reduced plasma vWF levels survived. Inhibition of Galpha12/alphaSNAP dependent vWF secretion may therefore be an effective strategy for blocking microvascular thrombosis, disseminated intravascular coagulation, and death due to sepsis.
Abstract 667: The Role of Aortic Wall-Nuclear Factor- kappa B (NF-kB) Signaling in Formation of Dissecting Aortic Aneurysms
Authors: Ijaz Talha , Sun Hong , Recinos Adrian , Tilton Ronald G , Brasier Allan R
Published: July 2021

Introduction:
Abdominal aortic aneurysm is a devastating disease since it can lead to aortic rupture and instantaneous death. We previously demonstrated that IL-6 secreted from the aortic wall is necessary for the development of abdominal aortic aneurysm and dissection (AAD). Since IL-6 is a NF-kB/RelA dependant gene, we investigated the role of aortic wall- NF-kB/RelA signaling in the development of AAD.

Methods and Results:
To test the role of aortic wall-RelA, we utilized Cre-Lox technology to delete RelA from aortic cells. Tamoxifen-inducible, Col1a2-promoter driven Cre mice (Col1a2-Cre) were crossed with mT/mG Cre-reporter mice to determine which aortic cells undergo genetic recombination after Cre activation. Flow cytometry analysis of the aortic wall indicated that 88% of the genetically recombined cells were SMCs and 8% were fibroblasts. Next, RelA floxed (RelA f/f) mice, generated in our lab, were crossed with Col1a2-Cre mice. RelA f/f Cre+ and RelA f/f Cre- were stimulated with tamoxifen for 10 days to generate aortic-RelA deficient (Ao-RelA-/-) or wild-type (Ao-RelA+/+) transgenics. Flow cytometry, qRT-PCR, and immunohistochemistry analysis suggested a depletion of aortic-RelA greater than 60%. To test the role of Ao-RelA in AAD, Ao-RelA -/- (n= 20) and Ao-RelA +/+ (n=14) mice were infused with angiotensin II for 7 days. Surprisingly, 20% of Ao-RelA-/- mice died from development of AAD and aortic rupture while no deaths were observed in Ao-RelA+/+ group. In addition, 40% of Ao-RelA-/- mice developed AAD compared to 14% of Ao-RelA+/+ mice. There was no significant difference in TUNEL staining or ERTR7+ fibroblast population between the two groups.

Conclusion:
Our studies suggest that aortic wall-RelA may be necessary for protection from AAD.
Abstract 681: The Effects of a Sustained-Release N-acetylcysteine Prodrug on Vascular Inflammation in Experimental Atherosclerosis
Authors: Kosuge Hisanori , Arad Yadon , Saito Toshinobu , Spin Joshua M , Isobe Mitsuaki , McConnell Michael V
Published: July 2021

Background:
Oxidative stress plays a key role in the development of atherosclerosis. N-acetylcysteine (NAC) has the potential to reduce oxidative stress and inflammation, but has a short half-life and low oral bioavailability. We studied a sustained-release formulation of NAC for the suppression of vascular inflammation in experimental atherosclerosis.

Methods:
ApoE-deficient mice (n=17) were started on a high-fat diet at the age of 7-13 weeks and given a daily oral gavage of either sustained-release NAC (n=8, 200mg/kg/mouse, total ~5040mg) or phosphate buffered saline (n=9) for 12 weeks. The sustained-release NAC formulation was an esterified prodrug called SURE-NACTM (Tiara Pharmaceuticals, Inc., Sunnyvale, CA). After 12 weeks, aortae were harvested and histology/immunohistochemistry performed. The levels of inflammatory cytokines in the plasma were evaluated using antibody-conjugated bead sets for 26 mouse cytokines and chemokines in multiplexed sandwich immunoassay format.

Results:
Lesion lipid area was significantly reduced in mice treated with sustained-release NAC compared to controls, for both the entire aorta and the aortic root: Lesion lipid area in entire aorta (% of total area) - 6.5±0.8% vs 13.3±1.1%, p<0.001; lesion lipid area in aortic root - 0.08±0.01mm2 vs 0.17±0.02mm2, p<0.001. Immunohistochemistry of the aortic root demonstrated that both macrophages and matrix metalloproteinase (MMP)-2 were also significantly suppressed by sustained-release NAC: Macrophages - 29.4±5.8% vs 55.6±3.2%, p<0.001; MMP-2 - 10.3±2.6% vs 19.2±2.9%, p<0.05. Inflammatory cytokines in mice treated with sustained-release NAC were significantly decreased compared to control (IL-2: 18.2%, IL-5: 47.6%, TNF-α: 34.2%, MIP-1α: 34.5% reduction). The levels of IL-2, IL-5, TNF-α, and MIP-1α showed a significant moderate correlation with both macrophage density and lipid area in the aortic sinus.

Conclusions:
A sustained-release NAC prodrug showed substantial suppression of lesion lipid area, macrophage infiltration, and inflammatory cytokines in experimental atherosclerosis. This may provide a novel daily oral therapy to reduce vascular inflammation for the prevention and treatment of atherosclerosis.
Abstract 684: Principal Component Analysis Identifies Lipoprotein Subfractions Associated with Coronary Artery Calcification in a Young High Cardiometabolic Risk Population: The Baptist Employee Healthy Heart Study
Authors: Veledar Emir , Aneni Ehimen , Osondu Chukwuemeka , Ogunmoroti Oluseye , Roberson Lare , Latif Muhammad , Jamal Omar , Ali Shozab S. , Aziz Muhammad , Younus Adnan , Ahmad Rameez , Khan Sher Ali , Post Janisse , Cury Ricardo , Feldman Theodore , Nasir Khurram
Published: July 2021

Introduction:
There is limited evidence relating lipoprotein subfractions or their composite measures to subclinical markers of atherosclerosis. In this study, we tested whether combinations of lipoprotein subfractions independently predict the presence of Coronary Artery Calcification (CAC) among presumably healthy people with high cardiovascular disease (CVD) risk.

Methods:
Cardio IQ™ Ion Mobility lipoprotein fractionation was measured at baseline in 172 high CVD risk participants of the Baptist Employee Healthy Heart Study (BEHHS). Principal component analysis was used to check for eventual components of CVD risk. Using bootstrap techniques, we created 1000 data sets of size 5000 and analyzed results of logistic regression.

Results:
Three principal components (PC) accounted for 88% variability. PC1 represented an increase in all lipoprotein measures; PC2 a decrease in HDL, large and medium LDL with an increase in VLDL and small and very small LDL; PC3 an increase in HDL, and a decrease in LDL subfractions and large IDL particles. The adjusted odds ratios for the presence of CAC were: 1.01(0.98 - 1.03) for PC1, 1.22(1.17-1.27) for PC2 and 0.90 (0.85 - 0.97) for PC3. PC2 and PC3 were associated with p-values smaller than 0.05 in 100% and 88% of all models.

Conclusion:
PCA of lipoprotein subfractions identified 3 independent components of CVD risk. PC2, proxy for the “atherogenic lipoprotein phenotype”, was consistently associated with the presence of underlying coronary artery calcification. There is potential to explore how lipoprotein subfractions as determined by a safe, inexpensive blood test can be used to stratify CVD risk in younger but high risk individuals.
Abstract 686: Intervention with Naringenin Enhances Weight Loss, Potentiates Improvements in Metabolic Dysregulation and Halts Progression of Atherosclerosis Induced by a High-Fat Diet in LDLr-/- Mice.
Authors: Burke Amy C , Sutherland Brian G , Assini Julia M , Huff Murray W
Published: July 2021

Previous studies demonstrate that the addition of naringenin, a grapefruit flavonoid, to a high-fat diet prevents the development of many disorders of the metabolic syndrome and atherosclerosis in Ldlr-/- mice. Furthermore, in intervention studies, the addition of naringenin to a high-fat, high cholesterol (HFHC) diet reversed pre-established obesity, hyperlipidemia, hepatic steatosis, insulin resistance and improved atherosclerotic lesion pathology, but not lesion size. In the present intervention study, we tested the hypothesis that addition of naringenin to a chow diet would further improve pre-established metabolic dysregulation and attenuate lesion development, compared to chow alone. Ldlr-/- mice were fed a HFHC diet for 12 weeks to induce metabolic dysregulation. Subsequently, mice received one of 3 diets for another 12 weeks: 1) continuation of the HFHC diet, 2) an isoflavone-free chow diet or 3) isoflavone-free chow with 3% naringenin. At 12 weeks, the HFHC diet induced significant weight gain and increased adiposity. Intervention with chow alone reduced the weight gained during induction by 22%, whereas the addition of naringenin to chow induced a weight loss of 71%. Specifically, the reduction in adiposity was 2.75-times greater in naringenin-treated mice, compared to chow alone. The HFHC diet increased VLDL cholesterol 20-fold and LDL cholesterol 5-fold, which were reduced by intervention with both chow (>60%) and chow supplemented with naringenin (>80%). The HFHC diet induced insulin resistance and glucose intolerance. Naringenin improved insulin tolerance (plasma glucose AUC -38%) and glucose tolerance (plasma glucose AUC -58%), which was accompanied by normalization of plasma insulin and glucose. HFHC-induction promoted the development of intermediate atherosclerotic lesions. Continuation of the HFHC diet doubled lesion size. Intervention with chow alone attenuated lesion size progression by 65%. The addition of naringenin to chow slowed lesion progression by 90%, resulting in smaller lesions compared to chow intervention alone (P=0.042). We conclude that intervention with naringenin-supplemented chow enhances weight loss, improves metabolic dysregulation and halts the progression of atherosclerosis.
Abstract 689: Cystathionine-Beta-Synthase Deficiency Induces Cardiac Hypertrophy and Contributes to Diminished Fatty Acid Oxidation in Mice with Diet-Induced Obesity
Authors: Glier Melissa B , Wambolt Rich B , Aleliuna Rika E , Gerrard Sarah L , da Silva Robin P , Ghosh Sanjoy , Lamers Yvonne , Jacobs Rene L , Allard Michael F , Devlin Angela M
Published: July 2021

Obesity-related cardiac lipid accumulation is associated with lipotoxicity and dysfunction. Cysteine is required for the synthesis of the antioxidant glutathione, which can be supplied by the transsulfuration of homocysteine by cystathionine-beta-synthase (Cbs). Cbs+/- mice with diet-induced obesity had greater glucose intolerance and lipotoxicity in the heart. Our objective was to determine the functional effects and mechanisms of cardiac lipotoxicity in Cbs+/- mice with diet-induced obesity. Cbs+/- and Cbs+/+ mice were fed a control diet or a high-fat diet (HFD) from weaning for 20 weeks. As expected, Cbs+/+ and Cbs+/- mice fed the HFD had greater final body weights, visceral (retroperitoneal and epididymal) and subcutaneous (inguinal) adiposity compared to mice fed the control diet. Cbs+/- mice had greater heart weights accompanied by higher concentrations of long chain polyunsaturated fatty acids arachidonic acid, 20:4n6 (AA) and docosahexaenoic acid, 22:6n3 (DHA) in the heart compared to Cbs+/+ mice. Mice fed the HFD had higher AA, but lower DHA concentrations in the heart compared to mice fed the control diet, with the greatest effect in Cbs+/- mice. Isolated working hearts revealed a reduced heart rate and cardiac output in Cbs+/- mice fed the control diet compared to Cbs+/+ mice. Independent of diet, Cbs+/- mice also had reduced aortic flow compared to Cbs+/+ mice, with a higher coronary flow only in Cbs+/- mice fed the HFD. Working hearts also revealed that Cbs+/- mice had lower palmitate oxidation rates compared to Cbs+/+ mice, with higher palmitate oxidation and glycolysis rates in mice fed the HFD compared to mice fed the control diet. Cbs+/- mice had a lower ratio of phosphorylated-AMP activated protein kinase alpha (AMPKα)/AMPKα expression (regulator of cellular energy) in heart compared to Cbs+/+ mice, and this occurred to a greater extent in those fed the HFD. Furthermore, we observed a higher ratio of collagen type 1(COL1α1)/ collagen type 3 (COL3α1) expression (implicated in myocardial stiffness) in heart, only in Cbs+/- mice fed the HFD. Collectively, these findings suggest that cardiac lipotoxicity in Cbs+/- mice with obesity is associated with cardiac hypertrophy, impaired cardiac fatty acid metabolism, and cardiac dysfunction.
Abstract 693: TLR3 Modulates Inflammation and Lipolysis in Visceral Adipose Tissue in High Fat Diet-fed Mice
Authors: Liu Cuiqing , Lin Guohua , Zhang Guoqing , Wang Huanhuan , Yin Hongping , Yu Liping , Sun Qinghua , Sun Lixian
Published: July 2021

Inflammation in insulin sensitive tissues, the visceral adipose tissue (VAT), is a central abnormality in obesity/insulin resistance (IR), with recruitment of innate immune cells such as monocytes into adipose tissue driving the development of glucose and lipoprotein dysregulation. We evaluated the role of Toll like receptor 3 (TLR3) in high fat diet-induced obesity and IR.

Wild-type C57BL/6 and TLR3
-/-
male mice were fed a high fat diet for 15 weeks. High fat feeding resulted in increased TLR3 expression in VAT. TLR3 deficiency attenuated the high fat diet-increased body weight, fasting blood glucose, whole body IR and impaired glucose tolerance. Morphologically, high fat diet induced adiposity and enlarged adipocyte area in VAT, which were attenuated in TLR3
-/-
mice. Functionally, high fat diet induced dysregulation of adipocytokines such as downregulation of adiponectin and resistin, upregulation of leptin in VAT, with the disturbance of adiponectin and leptin was corrected in TLR3-/- mice. In addition, high fat diet inhibited insulin pathway, accompanied with decreased phosphorylation of AMPK and lowered expression of lipolysis-related enzymes such as HSL and ATGL, both at the mRNA levels and protein levels, all of which was corrected by TLR3 deficiency. Finally, TLR3 deletion suppressed the high fat feeding-mediated macrophage polarization, evidenced by increased type M1 macrophage (F4/80+/CD11c+/CD206-) infiltration and upregulation of M1 genes such as IL-6 and TNFα. TLR3 modulates high fat diet-induced IR and obesity by suppressing M1 macrophage-mediated VAT inflammation, facilitating secretion of adipocyte-derived hormones, thus enhanced AMPK activity and adipose lipolysis. These findings provide new mechanistic links between dietary factors-mediated IR and associated abnormalities in lipid metabolism and adipose inflammation.
Abstract 7: LPA/PKD-1-HDAC7-FoxO1 Signaling-mediated Endothelial CD36 Transcriptional Repression and Proarteriogenic Reprogramming
Authors: Ren Bin , Best Brad , Ramakrishnan Devi , Walcott Brian , Storz Peter , Silverstein Roy
Published: July 2021

Background:
CD36 is a scavenger and antiangiogenic receptor that plays an important role in athero-thrombotic diseases, diabetes and cancer and contributes to obesity. Lysophosphatidic acid (LPA), a bioactive phospholipid signaling mediator, abolishes endothelial cell responses to antiangiogenic proteins containing thrombospondin type 1 homology domains by down-regulating endothelial CD36 transcription via protein kinase PKD-1 signaling. However, the precise mechanism as to how angiogenic signaling is integrated to regulate endothelial specific CD36 transcription remain unknown.

Hypothesis:
LPA represses CD36 transcription through PKD-1-mediated formation of a nuclear transcriptional complex in endothelial cells.

Methods:
Microvascular endothelial cells expressing CD36 were used for studying signaling and CD36 transcription by real time RT-qPCR, Western blotting, co-immunoprecipitation or avidin-biotin-conjugated DNA-binding assay; angiogenesis gene array was used for angiogenic gene profiling in response to LPA exposure. Spheroid-based angiogenesis assay, in vivo Matrigel assay and tumor angiogenesis model in CD36 deficiency and wild type mice were established to elucidate mechanisms of angiogenic signaling.

Results:
CD36 transcriptional repression involved PKD-1 signaling mediated formation of FoxO1-HDAC7 complex in the nucleus of endothelial cells. Unexpectedly, turning off CD36 transcription initiated reprogramming MVECs to express ephrin B2, a critical “molecular signature” involved in angiogenesis and arteriogenesis, and increased phosphorylation of Erk1/2, the MAP kinase important in arterial differentiation. PKD-1 signaling was also shown in tumor endothelium of Lewis lung carcinomas, along with low CD36 expression or CD36 deficiency. Angiogenic branching morphogenesis and in vivo angiogenesis were dependent on PKD-1 signaling.

Conclusion:
LPA/PKD1-HDAC7-FoxO1 signaling axis regulates endothelial CD36 transcription and mediates silencing of the antiangiogenic switch, resulting in proarteriogenic reprogramming. Targeting this signaling cascade could be a novel approach for cancer, diabetes, athero-thrombotic diseases and obesity.
Abstract 706: HDL and ApoA-I Inhibit Palmitate-induced Lipid Raft Trafficking of TLR4 and Inflammatory Cytokines Expression in Adipocytes
Authors: Yamada Hodaka , Umemoto Tomio , Ohtani Taeko , Fukaya Harue , Kawano Mikihiko , Kawakami Masanobu , Momomura Shinichi , Ishikawa Sane
Published: July 2021

Background:
Adipose tissue inflammation induces inflammatory molecules production leading to insulin resistance and increased cardiovascular disease risk. Recently it has become apparent that innate immune system is activated by saturated fatty acids (SFAs) by a process involved in TLR4 signaling. HDL and apoA-I has anti-inflammatory effects, however the mechanisms are not well understood in adipocytes.

Aims:
We examined to elucidate the effect of HDL and/or ApoA-I mediated cholesterol efflux on induction of inflammatory gene expression by palmitate in adipocytes via TLR4 dependent immune system.

Methods:
3T3-L1 adipocytes are exposure to 250uM palmitate for periods of up to 24 hours with or without HDL or Apo A-I. Antisense directed to ABCG-1 or ABCA-1 is added 24 hours prior to each reagent challenge. Cells were harvested and estimated for expression of
Saa
3 gene using real-time PCR or detection of TLR4 in raft or non raft by western blotting. Lipid rafts were purified by ultracentrifugation on discontinuous sucrose gradient, and identified by the presence of caveolin-1.

Results:
TLR4 was translocated into lipid raft from non raft within 15 minutes and dislocated by 60 minutes after palmitate exposure.
Saa
3 gene expression was significantly up-regulated by 24 hours palmitate exposure in adipocytes. TLR4 translocation to raft and
Saa
3 gene up- regulation induced by palmitate was significantly inhibited by pretreatment of HDL or Apo A-I for 6 hours. Antisense directed to ABCG-1 or SRB1 abrogated these inhibitory effects by HDL, but not ApoA-I whereas antisense to ABCA-1 did by Apo A-I but not HDL.

Conclusions:
These findings suggest that TLR4 mediates palmitate-induced
Saa
3 gene expression in adipocytes. A reduction in membrane cholesterol mediated by ABCG-1, SRB1 or ABCA-1 can account for the ability of HDL or ApoA-I to disrupt stimulation of inflammatory cytokines by palmitate
Abstract 710: A Novel Mechanism by Which CPI-17 Is Down-regulated in Cultured Vascular Smooth Muscle Cells and Tissues by TNFα and in C57BL/6 Mice by Lipopolysaccharide
Authors: Zhao Guogang , Liu Shu , Su Wen , Gong Ming C , Guo Zhenheng
Published: July 2021

Objectives:
Sepsis is characterized by a severe inflammatory response to infection, and its complications, including hypotension, can be fatal. It has long believed that hypotension in sepsis occur as a result of failure of the vascular smooth muscle cells (VSMC) to constrict. However, the molecular mechanism that links inflammation and decreases in vasoconstriction is largely unknown.

Approaches and Results:
CPI-17, a key regulator in vasoconstriction, was markedly decreased in mesentery arteries (MA) in mice injected with lipopolysaccharide (LPS). Incubation of cultured MA or VSMC with TNFα had similar effects on CPI-17 as LPS injection in mice. To identify transcriptional factors that respond to TNFα to suppress CPI-17 expression, we cloned a 1 kb murine CPI-17 promoter (-1,015 to +2 bp). Promoter deletion analysis revealed that a CPI-17 promoter region (-115 to -50 bp) was critical for TNFα to inhibit CPI-17 promoter activity. Bioinformatics analysis revealed this region contained 3 GC boxes that are putative binding sites for transcription factors SP1 and KLF4. Mutation of the 2nd and 3rd GC boxes, but not 1st GC box, abolished TNFα-mediated inhibitory effect on CPI-17 promoter activity. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) illustrated that SP1 and KLF4 bound to the 2nd and 3rd GC boxes, but not 1st GC box. Interestingly, TNFα had no effect on SP1 expression but suppressed SP1 binding to the CPI-17 promoter. In contrast, TNFα increased KLF4 expression and KLF4 binding to the CPI-17 promoter. Down-regulation of either SP1 or KLF4 by siRNA abolished inhibition of CPI-17 promoter activity by TNFα but the effects were opposite: SP1 siRNA decreased CPI-17 promoter activity whereas KLF4 siRNA increased CPI-17 promoter activity. Moreover, KLF4 up-regulation, but not SP1, was also found in MA in mice injected with LPS. KLF4 up-regulation by LPS preceded CPI-17 down-regulation and correlated with a decrease in vasoconstriction and hypotension.

Conclusions:
These studies reveal a novel mechanism by which CPI-17 is down-regulated by LPS/TNFα through SP1 and KLF4 in vascular wall and identify SP1, KLF4, and CPI-17 as new potential therapeutic targets for treatment of hypotension in patients with sepsis.
Abstract 717: An Association between Frontal Non-hemorrhagic Stroke and Atrial Fibrillation.
Authors: Sedhom Daniel , Yager Neil , Ashamalla Michael , Ghate Ketan , Pieper Justin , Nguyen Veronica , Shkolnik Boris , Torosoff Mikhail
Published: July 2021

Background:
Atrial fibrillation is a known risk factor for thromboembolic events. We sought to investigate an association between arrhythmia and stroke location in patients with non-hemorrhagic stroke.

Materials and methods:
The study cohort included 514 consecutive patients with non-hemorrhagic stroke treated at a single academic center. Stroke location was classified as frontal, occipital, lacunar, brainstem, left or right temporal, left or right parietal, cerebellar and insular. ANOVA, chi-square and logistic regression analyses were used. The study was approved by the institutional IRB.

Results:
Frontal embolic stroke was noted more frequently in patients with atrial fibrillation (29% vs. 15% in normal sinus rhythm, NSR, p<0.0001). No other anatomic location of the stroke was associated with arrhythmia.

Frontal strokes were more likely in patients with history of coronary artery disease (23% vs. 12%, p<0.005). However, there was no association between frontal stroke location and gender, age, history of hypertension, diabetes, dyslipidemia, peripheral vascular disease, or chronic renal insufficiency.

In multivariate logistic regression analysis, atrial fibrillation (HR 2.3; 95% CI 1.2-4.5, p=0.018) and history of coronary artery disease (HR 2; 95%CI 1.2-3.4, p=0.012) remained important predictors of frontal strokes.

Discussion:
Non-hemorrhagic strokes are common in patients with atrial fibrillation. We found increased prevalence of frontal embolic strokes in atrial fibrillation, possible due to the dual blood supply the frontal cerebrum from both the middle and anterior cerebral arteries. This association is intriguing and requires further studies.
Abstract 718: The Association of Erectile Dysfunction with Carotid Intima Media Thickness and Endothelial Dysfunction: A Meta-Analysis
Authors: Vo Bryan D , Aneni Ehimen , Oni Ebenezer , Maziak Wazim , Feldman Theodore , Fialkow Jonathan , Veledar Emir , Nasir Khurram , Feldman David , Blaha Michael
Published: July 2021

Introduction:
A recent meta-analysis showed that erectile dysfunction (ED) was associated with elevated cardiovascular disease (CVD) mortality. However, findings from studies examining the relationship between ED and sub-clinical CVD have been conflicting. This study summarizes the current evidence related to the association of ED and vascular disease as measured by flow mediated dilation (FMD) of the endothelium and carotid intima media thickness (CIMT).

Methods:
We searched multiple internet databases for published literature on studies assessing the association of ED and FMD, and CIMT between 1964 and 2014. A total of 11 studies met the inclusion criteria for examining the association between ED and FMD, while 7 studies met criteria for assessing the association with CIMT. Fixed-effect and random-effect models were used to assess and compare the standardized mean difference (SMD) of FMD and CIMT between persons with and without ED.

Results:
From a total of 795 individual participant data (590 with and 205 without ED), persons with ED had a 0.66mm (0.49, 0.83) increase in CIMT by fixed effects model or 0.67mm (0.44, 0.90) by random effects model compared to those without ED. Similarly, from 1055 individual participant data (536 with and 419 without ED), ED was associated with a significant reduction in FMD by both fixed effect (-1.10% (95% CI: -2.10, -0.95)) and random effect models (-1.53% (95% CI: -2.10, -0.95)). There was significant heterogeneity among studies assessing the relationship between ED and FMD (I2 = 93.2%, p<0.001) but not among those comparing CIMT among persons with ED (I2 = 43.5% p=0.09).

Conclusion:
This study confirms that ED is associated with both worsening endothelial function and increased CIMT. Thus, worsening vascular function may be a pathogenic mechanism for increased CVD morbidity and mortality in persons with ED. A diagnosis of ED should prompt a thorough CV work-up and aggressive preventive management.
Mid-Term Comparative Follow-Up After Aortic Valve Replacement with Carpentier-Edwards and Pericarbon Pericardial Prostheses
Authors: Le Tourneau Thierry , Savoye Christine , McFadden Eugene P. , Grandmougin Daniel , Carton Hubert-François , Hennequin Jean-Luc , Dubar Arnaud , Fayad Georges , Warembourg Henri
Published: July 2021

Background
—The first generation of pericardial valves had a high rate of premature deterioration. The aim of this study was to compare the outcome after aortic valve replacement with second generation pericardial prostheses (Pericarbon and Carpentier-Edwards).

Methods and Results
—Between 1987 and 1994, 162 patients underwent aortic valve replacement with either a Pericarbon (n=81, 69±11 years) or a Carpentier-Edwards (n=81, 70±11 years) pericardial prosthesis. Mean follow-up was 4.4±2.7 years for Pericarbon and 4.8±2.4 years for Carpentier-Edwards valves (
P
=0.27), giving a total follow-up of 745 patient-years. Thirty-day mortality and 5-year actuarial survival were, respectively, 6.2% and 63.2±5.7% in the Pericarbon group and 6.2% and 63.5±5.6% in the Carpentier-Edwards group. At 8 years, freedom from (and linearized rates per patient-year) thromboembolism, structural failure, and all valve-related events were, respectively, 91.8±3.6% (1.4%), 76.9±8.7% (2.5%), and 58.4±9.3% (5.6%) in the Pericarbon group and 94.4±2.7% (1%), 100% (0%,
P
<0.01), and 88.8±3.7% (2%,
P
<0.05) in the Carpentier-Edwards group. There were 9 (11.1%) Pericarbon structural failures related predominantly to severe calcification and stenosis. The actual reoperation rate was 7.4% (1.6% per patient-year) in the Pericarbon group for fibrocalcific degeneration (n=3), periprosthetic leak (n=1), endocarditis (n=1), and aortic dissection (n=1). There was neither structural valve failure nor valve reoperation in the Carpentier-Edwards group. Echocardiographic review of 70 patients from 85 survivors (82.3%) found 4 additional Pericarbon valves with signs of early structural failure but no Carpentier-Edwards valve with such changes.

Conclusions
—Eight years after aortic valve replacement, Pericarbon pericardial prostheses compared unfavorably with Carpentier-Edwards pericardial prostheses, with a high incidence of structural valve failure and reoperation.