bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 279: Role of PDE10A in Arterial Calcification
Authors: Cai Yujun , Wang Xue-lin , Lin Tonghui , Guzman Raul J
Published: March 2019

Vascular calcification is highly prevalent in patients with diabetes mellitus and chronic kidney disease. When located in the media, arterial calcification is strongly associated with increased cardiovascular morbidity and mortality. The second messenger cyclic nucleotides cAMP and cGMP, controlled by distinct cyclic nucleotide phosphodiesterase (PDE) isozymes, play important regulatory roles in a variety of human diseases. Using a qPCR PDE array, we found that PDE10A was the most highly induced among all PDE genes in a rat model of medial artery calcification. PDE10A expression was markedly increased in calcified arteries from rats with chronic kidney disease and in tibial arteries from patients with peripheral artery disease. Interestingly, it co-localized with osteogenic markers in these specimens.
In vitro
, PDE10A knockdown using siRNA, and inhibition with a synthetic inhibitor markedly reduced osteogenic transformation and calcification of vascular SMC exposed to high phosphate levels. Aortic rings from PDE10A knockout mice showed significantly less Pi-induced medial calcification than those from wild-type controls. Deficiency of PDE10A also reduced medial calcification in a mouse medial calcification model
in vivo
. Mechanistic studies to elucidate the signaling alterations invoked by PDE10A are ongoing. These findings suggest that PDE10A plays a crucial role in the development of medial artery calcification, and that targeting it may provide a novel therapeutic strategy for reducing medial calcification and improving outcomes in patients with PAD.
Abstract 280: Are There Sex-specific Differences in Arteriovenous Fistula Maturation?
Authors: Kudze Tambudzai , Isaji Toshihiko , Ono Shun , Hashimoto Takuya , Yatsula Bogdan , Hu Haidi , Liu Haiyang , Dardik Alan
Published: March 2019

The arteriovenous fistula (AVF) is the preferred method of dialysis access due to its proven superior long term outcomes. However, women have lower rates of AVF maturation than men (38% vs. 60%), preventing optimal AVF use. Using a novel mouse AVF model that recapitulates human AVF maturation, we tested the hypothesis that there is a difference in male and female AVF maturation.

Aortocaval fistulae were created in male and female C57BL/6 mice (9-10wk). At days 0, 3, 7, 14 and 21, aortic and IVC diameters and flow velocity were monitored by Doppler ultrasound. We then calculated shear stress. Using qPCR, we measured messenger RNA (mRNA). AVF were examined at day 21 and AVF wall thickness was measured by computer morphometry.

Female mice weighed less preoperatively and at day 21 (p<0.05). They also had larger suprarenal aortic diameter (p=0.002) but smaller infrarenal IVC diameter (p<0.05) at baseline. After AVF creation, there was similar dilation of the infrarenal aorta and IVC in both male and female mice (p>0.05). Infrarenal IVC mean velocity was decreased in female mice at baseline and at day 3, 14 and 21 (p<0.05). Similarly, mean laminar shear stress magnitude in the infrarenal IVC was decreased in female mice at day 7 (p=0.03), 14 (p=0.04) and 21 (p=0.01). There was no difference in the infrarenal aorta shear stress magnitude (p=0.80). mRNA of KLF2, a marker of laminar shear stress, was decreased in the venous limb of female AVF on day 21 (p=0.048). Preoperatively, female mice had thinner venous walls (p=0.035). However, at day 21, AVF wall thickness was similar (p=0.18).

AVF in female mice have lower magnitudes of laminar shear stress, lower expression of KLF2 mRNA and a higher percent increase in wall thickness. These findings suggest a mechanism underlying the diminished rates of AVF maturation in women.
Abstract 287: Transgenic Overexpression of Dimethylarginine Dimethylaminohydrolase 1 Protects From Angiotensin II - Induced Cardiac Hypertrophy and Vascular Remodeling
Authors: Kopaliani Irakli , Jarzebska Natalia , Brilloff Silke , Kolouschek Anne , Martens-Lobenhoffer Jens , Bode-Böger Stefanie M , Deussen Andreas , Weiss Norbert , Rodionov Roman N
Published: March 2019

Background:
Dimethylarginine dimethylaminohydrolase 1 (DDAH1) hydrolyzes the endogenous inhibitor of nitric oxide synthases asymmetric dimethylarginine (ADMA). DDAH1 is also suggested to have ADMA-independent effects. DDAH1 overexpression lowers ADMA levels and protects from renal interstitial fibrosis and vascular oxidative stress in angiotensin-II-induced hypertension. The current study was designed to test the hypothesis that DDAH1 overexpression protects from angiotensin II-induced cardiac hypertrophy and vascular remodeling.

Methods:
Angiotensin II (AngII) was infused in the doses of 0.75 and 1.5 mg/kg/day, respectively, in DDAH1 transgenic mice (TG) and wild type (WT) littermates via osmotic minipumps. Echocardiography was performed in the first and fourth week after start of the infusion. Systolic blood pressure was measured by the tail-cuff method. Cardiac hypertrophy and vascular remodeling was assessed by histology after 4 weeks of AngII infusion.

Results:
TG mice had decreased plasma and tissue ADMA. Infusion of Ang II resulted in an increase in systolic blood pressure, which was similar between TG and WT mice at week 1, however, TG mice were protected from a further increase in blood pressure. After 4-weeks infusion of AngII TG mice had significantly higher left ventricular lumen to wall ratio, smaller size of cardiomyocytes and reduced myocardial collagen expression compared to WT littermates. TG mice had lower left ventricular posterior wall thickness in systole and diastole as compared to WT controls. The vasomotor function of aortic rings in response to acetylcholine was improved in the TG mice as compared to the WT mice. TG mice had less aortic hypertrophy and fibrosis and more elastin in aorta as compared to WT mice. Aortic infiltration of CD45
+
, CD3
+
, CD8
+
and CD4
+
T-cells was significantly lower in TG than in WT mice.

Conclusion:
This study shows that upregulation of DDAH1 protects from AngII-induced cardiac hypertrophy and vascular remodeling. Upregulation of DDAH1 might be a potential therapeutic approach for protection from AngII – induced end organ damage. We are currently investigating, whether protective effects of DDAH1 are ADMA-dependent or ADMA-independent.
Abstract 301: A New Model of Murine Stasis Pulmonary Thromboembolism in vivo With Assessment by Noninvasive Multimodal Molecular-Structural Imaging
Authors: Kessinger Chase W , Jaffer Farouc A
Published: March 2019

Objective:
Pulmonary embolism (PE) is a life-threatening cardiovascular disease that urgently requires improved diagnostic, prognostic and therapeutic options. Here we established and investigated a novel murine PE model based on embolization of a stasis-induced venous thrombus (VT). We further utilized in vivo molecular-structural imaging of fibrin accessibility to assess the healing status of the PE.

Methods:
Stasis VT was induced in the femoral veins of donor C57BL6 mice. VT was resected 24 hours (h) after induction and then intravenously injected into recipient mice to create PE. Pulse-wave Doppler of pulmonary artery flow was utilized to evaluate the pulmonary arterial pressure (PAP) before and 24h after embolization. One day after embolization, the fibrin-specific probe FTP11-CyAm7 (750/767nm ex/em, 150 nmoles/kg i.v.) assessed the FTP binding and spatial localization of PE via integrated fluorescence-mediated tomography and computed tomography (FMT-CT) pulmonary angiography. Mice were sacrificed 1, 4, 8, or 21 days following stasis-VT embolization, and underwent histological analysis of the PE.

Results:
Femoral vein stasis-VT exhibited similar temporal inflammatory neutrophil and monocyte/macrophage infiltration and FTP11 binding (fibrin accessibility) kinetics compared to inferior vena cava stasis-VT models. The mean VT width and length were 2.4±0.2mm and 0.65±0.02mm, respectively (n=29 thrombi). Noninvasive ultrasound assessment of the mouse PAP, assessed via the pulmonary acceleration time (PAT), showed a 22% decrease (p=0.03) in pre-to-post-PE PAT times, consistent with increased PAP due to PE. FMT-CT imaging using FTP11 demonstrated accessible fibrin in day 1 PE and visualized PE confined to the lungs, primarily (91%) in the right lung. Ex vivo fluorescence reflectance imaging and histological analysis verified the location PE reported by FMT-CT.

Conclusions:
These data validate the first in vivo model of PE based on stasis VT formed in vivo. The model reproducibly generates a survival PE that harbors accessible fibrin in recent stasis VT lodged in the lung, and acute increases PAP. This new clinically relevant model of PE is anticipated to facilitate research of acute PE and chronic thromboembolic pulmonary hypertension.
Abstract 331: Fatty Acid Synthase Expression in Peripheral Arterial Plaque
Authors: Desai Kshitij A , DeSilva Gayan , Chao Yang , Sanchez Luis , Semenkovich Clay , Zayed Mohamed
Published: March 2019

Introduction:
Diabetic patients exhibit distinct patterns of diffuse and recalcitrant peripheral artery disease, which subsequently predisposes them to poor wound-healing, infection, and critical limb ischemia. Fatty Acid Synthase (FAS), an enzyme responsible for de novo fatty acid synthesis, exhibits elevated content in the liver tissue in diabetic mouse models and elevated serum levels in diabetic subjects with carotid artery stenosis. It is currently unknown whether tissue FAS and serum circulating FAS (cFAS) correlate with diabetic status in patients with femoral arterial- occlusive disease.

Materials and Methods:
We enrolled diabetic and non-diabetic subjects undergoing femoral endarterectomy (FEA) in an IRB-approved vascular biobank. Serum and femoral endarterectomy plaque samples were collected from participants, and tissue FAS and serum cFAS, content was evaluated using ELISA. Differences in cFAS content between cohorts were summarized as mean ± SEM, and statistical significance was determined via T-test and Pearson correlation analysis.

Results:
33 patients (16 DM, 17 non-DM) who underwent femoral endarterectomy for high grade occlusive disease were evaluated. No significant difference in key demographics were observed. Tissue plaque FAS content was 69.8% (13.3 vs 7.81 ELISA: Total protein) higher in DM compared to non-DM subjects (p-value = 0.011). cFAS was also elevated by 41.7% (1.39 vs 0.98 ELISA: Total protein) in diabetic compared to non-DM subjects (p-value = 0.048). Correlation analysis of 23 patients’ paired samples revealed a significant correlation between cFAS and plaque FAS content (Pearson r = 0.47, p-vale = 0.023).

Conclusion:
Our study is the first to evaluate cFAS levels in patients with high grade, symptomatic, lower extremity PAD, and demonstrates evidence that cFAS and tissue FAS levels correlate in subjects with diabetes. Future studies will help determine whether cFAS is a relevant biomarker for disease severity and progression in diabetic subjects.
Abstract 335: Phase Contrast Magnetic Resonance Imaging as a Novel Complementary Tool for Investigations of Peripheral Artery Disease
Authors: Cao Amos A , Scheven Ulrich M , Greve Joan M
Published: March 2019

Therapeutic angiogenesis is pertinent to the acute survival of distal ischemic tissue and a relevant treatment for critical limb ischemia (CLI). Therapeutic arteriogeneis is pertinent to the restoration of function, via collateral vessel formation in proximal tissue that is not typically ischemic, and a relevant treatment for preventing intermittent claudication from advancing to CLI. In preclinical models of peripheral artery disease (PAD), laser perfusion imaging methods are applicable to the former. However, recently published results indicate they underestimate recovery and their limited penetration depth restrict their application to the latter. Our objective was to evaluate whether phase contrast magnetic resonance imaging (PC-MRI) could be used as a complementary method in murine models of PAD.

C57BL/6 male mice underwent femoral artery ligation (n=5). Mice were imaged prior to and day 3 after surgery. PC-MRI uses the phase of the MRI signal to calculate velocity of blood flow within a voxel. Integrating velocity across the vessel lumen results in flow measurements. Maximum velocity and mean volumetric flow were compared at the infrarenal aorta and ipsilateral (Ip) and contralateral (Con) iliac arteries.

Maximum velocities did not change in the aorta or Con (pre vs. day 3: aorta, 55.4±7.2 vs. 48.9±9.4 cm/sec; Con, 45.7±7.5 vs. 40.7±8.0 cm/sec). Mean volumetric flows also did not change in the aorta or Con (pre vs. day 3: aorta, 2.0±0.7 vs. 1.53±0.4 mL/min; Con, 1.1±0.4 vs. 0.92±0.3 mL/min). In the ipsilateral iliac artery, maximum velocity was reduced by ~35% (pre vs. day 3: 43.6±5.7 vs. 28.4±5.5 cm/sec, p=0.002), leading to a reduction in mean flow of ~41% (1.1±0.4 vs. 0.63±0.3 mL/min, p=0.04). Ip/Con ratios for both metrics were: 0.98±0.2 vs. 0.74±0.2 cm/sec, p=0.03; and, 1.0±0.1 vs. 0.72±0.3 mL/min, p=0.03.

Our previous work demonstrated the ability to promote proximal-arteriogenesis by inducing distal-angiogenesis via application of a slow-release growth factor. Having developed a non-invasive method to directly quantify velocity and flow in large proximal vessels, we will have the opportunity to explore the biomechanical mechanisms coupling angio- and arterio-genesis even though they occur temporally and spatially separated.
Abstract 340: Microvascular Pathology in Peripheral Artery Disease
Authors: Mietus Constance J , Lackner Timothy , Karvelis Peter , Lambert Nicholas , Hernandez Hernan , Pipinos Iraklis , Casale George
Published: March 2019

Background:
Peripheral Artery Disease (PAD) is caused by atherosclerotic narrowing of arteries supplying the legs. PAD-induced myopathy is characterized by myofiber degeneration and progressive fibrosis. Qualitative histological review suggests pathological changes in the microvasculature of PAD muscle, in association with advancing fibrosis. We tested the hypothesis that microvessel architecture, pericyte coverage, and collagen profiles systematically change with advancing disease and are consistent with advancing microvascular pathology.

Methods:
Biopsies of PAD patients at Fontaine Stage II (n=15) and IV (n=16), and controls (n=15) were labeled with antibodies specific for Col I, Col IV, αSMA, or CD31 and analyzed by quantitative wide-field, fluorescence microscopy. Thickness of the basement membrane (BM), peri-microvascular Col I density, and BM lumen diameter were measured. Pericytes were identified by abluminal location within the microvascular BM and αSMA
+
labeling. Group differences were tested by ANOVA and a
post hoc
pairwise T Test with Bonferroni correction. Correlations were determined by linear regression analysis.

Results:
Thickness of the BM was greater in Stage II patients (1.58 μm) compared to controls (1.42 μm) (p<0.043) and in Stage IV (1.75 μm) compared to Stage II patients (p<0.021). Microvascular BM lumen diameter was increased (p<0.001) in Stage IV patients (3.97 μm) compared to control (3.25 μm) and Stage II patients (3.29 μm). Thickened PAD microvessels had greater pericyte coverage than control microvessels. BM thickness correlated positively with microvascular BM lumen diameter (R
2
= 0.513, p<0.001). Peri-microvascular Col I deposition correlated positively with microvascular lumen diameter (R
2
= 0.162, p=0.006), and was greater in Stage IV compared to Stage II patients (p=0.040).

Conclusions:
Increased perivascular Col I deposition, BM thickening, and BM lumen diameter represent advancing microvascular disease in PAD patients. Pericytes, which deposit BM collagen, are more abundant in thickened microvessels. Pericyte replication and secretion of Col IV may be determining factors in the microvascular pathology of PAD muscle.
Abstract 343: Baseline Serum Uric acid Predicts Future Cardiovascular Events and Death in Patients Undergoing Carotid Endarterectomy
Authors: Prasad Megha , Nardi Valentina , Meyer Fred , Lerman Lilach , Lerman Amir
Published: March 2019

Background:
Serum uric acid is a marker of oxidative stress, and may serve as a marker of adverse cardiovascular events and outcomes including cardiovascular death, hospitalization and myocardial infarction. We aimed to determine whether uric acid predicts cardiovascular events and death in patients undergoing carotid endarterectomy (CEA).

Methods:
We enrolled 327 patients undergoing CEA between February 2002 and June 2017 who had complete laboratory work up at baseline. Patients were followed through the electronic chart as well as via regular nursing follow-up for a median of 10.9 years (IQR 7.3, 13.4). There were a total of 67 adverse cardiovascular events during follow-up.

Results:
Baseline serum uric acid levels were significantly higher in patients who developed cardiovascular events during the follow-up period vs. those that did not [6.87
+
2.29 vs. 6.39
+
1.61); p=0.047). Baseline serum uric was independently associated with cardiovascular events when adjusting for age, creatinine, diabetes mellitus, and hyperlipidemia (L-R 4.0, p=0.04)(Figure 1). Sex stratified analysis showed that serum uric acid was also an independent predictor of both cardiovascular events and death in men, not women undergoing after adjustment for confounders including age, cholesterol, systolic blood pressure and glucose (LR 4.97; p=0.03).

Discussion:
Higher uric acid levels are associated with cardiovascular events and death after CEA. Further investigation is necessary to determine the role of uric acid as a modifiable risk factor in these patients.
Abstract 348: The Blood-Pressure-Lowering Effect of Moxonidine May Be Partially Through the Prostaglandin Pathway in the Kidney
Authors: Wang Yutang , Steicke Michelle , Golledge Jonathan
Published: March 2019

Objectives:
Moxonidine is believed to decrease blood pressure via inhibiting sympathetic activity in the brain and consequently decreasing vascular tone. However, whether direct peripheral mechanisms are involved in this process is unknown. This study aimed to investigate possible peripheral mechanisms of moxonidine in lowering blood pressure.

Methods:
Two groups (N = 10 in each group) of apolipoprotein E-deficient mice were used. These animals were treated with or without moxonidine (18 mg/kg body weight per day) via drinking water throughout the experiment. Three days after the initiation of moxonidine treatment, hypertension was induced by subcutaneous infusion of angiotensin II (1 μg/kg body weight per min) for 28 days. Blood pressure was measured by the tail cuff method. Size and area of kidney tubules and glomeruli were analyzed by morphometric analysis. Gene expression was assessed by quantitative PCR.

Results:
Moxonidine treatment significantly decreased blood pressure (P<0.05). It did not change the size or area of either kidney tubules or glomeruli (P>0.05), nor the renal expression of inflammatory markers. Both types of moxonidine receptors (imidazoline 1 and alpha-2 adrenergic receptors) were found to be expressed in the kidney, and their gene expression was not altered by moxonidine treatment. However, moxonidine treatment significantly increased the expression of prostaglandin EP3 receptor (P<0.05). Increased prostaglandin EP3 receptor has been reported to be associated with promoting diuresis and natriuresis and thus decreasing blood pressure.

Summary:
The results indicate that moxonidine may lower blood pressure partially through an unrecognized non-central pathway by upregulating prostaglandin EP3 receptor in the kidney in mice.
Abstract 367: Quantitative Phosphoproteomic Profiling and Causal Pathway Analysis Reveal Novel Mediators and Mechanisms Supporting Glycoprotein VI (GPVI) Signaling and Platelet Function
Authors: Babur Özgün , Ngo Anh T , Zilberman-Rudenko Jevgenia , McCarty Owen J , Demir Emek , David Larry L , Aslan Joseph E
Published: March 2019

Intracellular signaling systems associated with the platelet glycoprotein VI receptor (GPVI) are essential to hemostasis and also initiate atherothrombotic events in cardiovascular disease contexts. While many separable molecular entities linked to GPVI-mediated platelet activation have been characterized, a systemic understanding of GPVI signaling over the context of the entire platelet proteome remains far from complete. To test the hypothesis that specific, intact signaling systems downstream of GPVI regulate platelet function, we used a combination of tandem mass tag (TMT) labeling and high-resolution, multidimensional mass spectrometry and informatics tools to quantify and model protein phosphorylation events reproducibly measurable in platelets from healthy human subjects (n=5) in response to the GPVI specific agonist collagen-related peptide (CRP). Following CRP treatment, platelets rapidly regulated the phosphorylation state of 1004 serine, threonine and tyrosine residues on 542 proteins (fold change > 1.5; false discovery rate < 0.05). In addition to regulatory sites on kinases, phospholipases and other well-known targets, the majority of phosphorylation events following CRP stimulation occurred on proteins with uncharacterized roles in platelet function. CausalPath analysis of this data mapped 46 high confidence pathway relations, suggesting that Syk, PKCs and MAPKs ultimately drive the phosphorylation of several novel effectors specifically downstream of GPVI in platelet regulation, including CRMP-2, RAD23B and caldesmon. In addition to comprehensively mapping out known signaling relations while illuminating previously unrecognized regulatory mechanisms of platelet function around GPVI, our work provides a platform to generate novel, mechanistic and testable hypotheses from omics data sets for subsequent discovery-driven efforts to identify platelet-based therapeutic and biomarker targets relevant to cardiovascular disease risk, progression and mortality.
Abstract 369: Muscle Ischemia Induces Nlrp3 Inflammasome Activation in Platelets via Tlr4, Promoting Platelet Aggregation and Interfering With Perfusion Recovery
Authors: Vogel Sebastian , Murthy Pranav , Cui Xiangdong , Xie Bowen , Sachdev Ulka
Published: March 2019

Introduction:
Angiogenesis is an adaptive response to chronic ischemia, but is deficient in peripheral arterial disease (PAD). We have recently shown that the pattern recognition receptors toll-like receptor 4 (TLR4) and nucleotide-binding domain leucine rich repeat containing protein 3 (NLRP3) expressed by platelets control aggregation and thrombosis. The role of platelet TLR4 and NLRP3 in PAD is unexplored.

Methods:
Unilateral femoral artery ligation (FAL) was performed in transgenic mice with platelet-specific ablation of TLR4 (TLR4 PF4) and in global NLRP3 knockout (NLRP3 KO) mice. Platelet NLRP3 inflammasome activation was monitored by caspase-1 activation (fluorescent labeled inhibitor of caspase-1, FLICA) and cleavage of IL1β (Western blot). Platelet aggregation was evaluated with impedance aggregometry. Laser Doppler perfusion imaging (LDPI) verified perfusion in the ischemic (right) and non-ischemic (left) limb over time. Angiogenesis and myoblast regeneration were measured histologically.

Results:
Platelet NLRP3 inflammasome activity was significantly upregulated following FAL (p<0.001), and reversed with TAK242, a TLR4 inhibitor. FAL significantly increased aggregation of circulating platelets, which was significantly suppressed in TLR4 PF4 (p<0.01) and NLRP3 KO mice (p<0.001). Down-regulation of platelet aggregation and caspase-1 activity in TLR4 PF4 mice was nearly completely reversed by nigericin, a NLRP3 activator. Ischemic limb perfusion (Fig 1A) was significantly higher in TLR4 PF4 (Fig1B, p<0.05) and NLRP3 KO mice (Fig1C, p<0.001) than in controls 14d after FAL. Angiogenesis and regeneration were significantly improved in TLR4 PF4 mice (p<0.05).

Conclusion:
We show that the platelet NLRP3 inflammasome is activated in muscle ischemia via platelet TLR4, which upregulates platelet aggregation and deters recovery from FAL. Thus, platelet TRL4/NLRP3 activation may be a therapeutic target to improve limb salvage in PAD.
Abstract 373: An Animal Model of Left Atrial Thrombus
Authors: Chen Jiqiu , Ceholski Delaine , Liang Lifan , Hajjar Roger
Published: March 2019

Left atrial thrombus (LAT) is a common funding of some cardiovascular diseases such as congestive heart failure (CHF), atrial fibrillation and myocardial injury. There is no proper animal model of LAT to date. The present study reports a model of congestive heart failure in rats that is characterized by LAT.

Methods:
LAT were induced by aortic banding 2 months plus 30 minutes ischemia and reperfusion 1 m followed with aortic de-banding 1 m in rats. Cardiac function and blood flow velocity were assessed by echocardiography. Masson’s stainings were conducted for histology.

Results:
congestive heart failure (CHF) in rats with LAT showed significant dilatation in left atrium (CHF = 56 ± 18 mm, N = 7, Control = 25 ± 6 mm, N = 10), significant decrease in left appendage blood flow velocity (CHF = 4.20 ± 0.09, Control = 0.21 ± 0.05 m/s) and mitral valve blood velocity (CHF = 0.76 ± 0.09 m/s, Control = 1.01 ± 0.04 m/s). Histology data showed that mature LAT had a complete membrane; the thrombus was characterized by significant fibrosis in middle and out layers. The outer layer of thrombus had microvessels and the cells were nucleated. The central part of the thrombus had very little fibrosis and showed necrosis. Small thrombus (0.05 - 0.1 mm) had a base from which the blood clot grew. Atrial neo-intimal fibrosis plays an important role in the formation of the base of microembolization. However, in some cases, micro-thrombi (<0.01 mm) showed no fibrotic base.

Conclusion:
left atrial thrombus results from blood flow congestion. However, it is still not clear whether the thrombus is due to vascular proliferation from the atrial wall or through adhesion of coagulation fibers to the atrial wall.

Figure. A. echocardiographic imaging. B. left atrial area . C. mitral valve blood velocity. D. echocardiography of left atrial blood velocity. E. left atrial blood flow velocity. F. regular imaging of LAT. G. cross sectional imaging. H. small LAT. I. endometrial fibrosis. J. micro thrombus. K. coagulation fiber.
Abstract 390: A New Humanized Mouse Model to Study Lipid Abnormalities and HDL Function in Poorly Controlled Type 1 Diabetes Mellitus
Authors: Kothari Vishal , He Yi , Kramer Farah , Barnhart Shelley , Kanter Jenny E , Chait Alan , Goldberg Ira J , Vaisar Tomas , Heinecke Jay W , Bornfeldt Karin E
Published: March 2019

Elevated plasma triglycerides (TG) and VLDL are often present in poorly controlled type 1 diabetes mellitus (T1DM). Furthermore, T1DM patients exhibit an increased risk of cardiovascular disease (CVD) despite normal or high HDL-cholesterol. There is a need for new mouse models to study T1DM lipid abnormalities because mice do not express cholesteryl ester transfer protein (CETP), which transfers cholesteryl ester (CE) from HDL to VLDL and other lipoproteins in exchange for TG, and because mice do not exhibit human-like HDL populations. To develop a physiologically relevant model, we crossed a virally-induced mouse model of T1DM (

Ldlr
-/-
;Gp
Tg

) deficient in the LDL receptor with human apolipoprotein A-I (
APOA1
) transgenic mice. These mice (n=8-10) were injected with a liver-specific adeno-associated virus driving expression of human CETP, mimicking the human CETP activity/apoA-I ratio, and the 3 major human HDL subpopulations. Using this model (

APOA1
Tg
;Ldlr
-/-
;Gp
Tg

), we examined the impact of diabetes on dyslipidemia, HDL particle concentration (HDL-P), and serum HDL function. Induction of diabetes increased plasma TG and cholesterol, primarily due to an increase in VLDL, as compared with non-diabetic (ND) littermates. The dyslipidemia was associated with increases in hepatic TG content and hepatic TG production rate (2-fold over ND). Moreover, resident peritoneal macrophages from diabetic mice exhibited a 5-fold increase in CE loading, increased lipid raft content (20%), and increased
Tnfa
(3-fold) and
Il1b
mRNA (4-fold), as compared with ND littermates. In addition, T1DM mice had increased small, medium and large HDL-P (2-fold) concentrations compared with ND controls. Serum HDL from T1DM mice showed a significant increase in its cholesterol efflux capacity through both ABCA1 and ABCG1. Thus, in this humanized mouse model, poorly controlled T1DM leads to elevated plasma TG and VLDL due in part to increased hepatic TG production. This dyslipidemia is associated with higher HDL-P and an increase in serum HDL’s cholesterol efflux capacity, likely due to the increased plasma HDL-P. We have found similar changes in HDL concentration and function in humans with T1DM, suggesting that our mouse model might provide insights into CVD risk in T1DM patients.
Abstract 391: Flow Field-flow Fractionation: a Powerful Underused Size Fractionation Technique for Composition Analysis of Lipoprotein Particles
Authors: Kuklenyik Zsuzsanna , Bierboum Kevin , Jones Jeffrey , Williamson Yulanda , Barr John R
Published: March 2019

Multiplexed mass spectrometry (MS) techniques have substantially improved the ability to characterize the composition of lipoproteins. However, these advanced MS techniques for composition analysis of lipoproteins are limited by traditional preparative fractionation techniques that compromise the structural integrity of lipoprotein particles during separation from serum or plasma. In this work a highly effective but underused size fractionation technique, asymmetric flow field-flow fractionation (AFFFF) was applied. We compared AFFFF with traditional techniques, including gel electrophoresis (GE), size exclusion chromatography (SEC), and gradient ultracentrifugation (GUC). Direct comparison of the techniques was performed by analysis of the same set of samples using sudan black staining. The AFFFF size separation allowed size resolution most similar to GE techniques, either tube gel or gradient gel. In addition, AFFFF and SEC was compared by collection of fractions where the average size in each fraction was measured by dynamic light scattering, and all major lipids and apolipoproteins were quantified by liquid chromatography tandem mass spectrometry methods (LC-MS/MS) in each fraction. The size resolution by AFFFF vs. SEC was compared based on the elution profiles of endogenous plasma proteins. On the hydrodynamic size scale, the half peak width of individual size species were 0.8-1 nm by AFFFF vs. 3-8 nm by SEC, showing the superior size resolution capability of the AFFFF technique. The gentle nature of the AFFFF separation relative to GUC was demonstrated by AFFFF separation of HDL sub-fractions with and without pre-fractionation by GUC, showing the significant loss of apoA-IV and apoCs, as well as a general shift of all lipid and protein profiles to smaller hydrodynamic size as a result of intense shearing forces and high salt conditions during GUC. Our results demonstrate that AFFFF coupled with LC-MS/MS deserves to be among the gold standard techniques for size and composition analysis of HDL, LDL and VLDL sub-classes.
Abstract 393: Differential Regulation of Lipoprotein Assembly and Secretion and Mtp Expression in Intestinal and Hepatic Cells by Oleoylethanolamide
Authors: Pan Xiaoyue , Hussain Mahmood M
Published: March 2019

Dietary fat triggers synthesis of OEA in enterocytes of the upper small intestine. OEA initiates a cascade of events to optimize fatty acid uptake and reduce feeding frequency. Effects of OEA have been studied after oral gavage and intraperitoneal injections. Here, we studied dose dependent effects of OEA on the assembly and secretion of lipoproteins in differentiated human colon carcinoma Caco-2, human hepatoma Huh-7 and HepG2 cells as well as in mouse primary enterocytes and hepatocytes. Differentiated Caco-2 cells were treated with OEA or oleic acid for 16 h. OEA increased synthesis of triglycerides in dose dependent manner compared with oleic acid:taurocholate micelles. Further, OEA increased secretion of triglycerides and apolipoprotein B (apoB) in chylomicrons. Expression and activity of microsomal triglyceride transfer protein (MTP) was increased by OEA in a dose dependent manner in these cells. Next, mouse primary enterocytes were incubated with OEA or oleic acid for 4 h. OEA increased MTP activity and secretion of triglycerides in these cells as well. These studies suggest that OEA enhances MTP expression, lipid synthesis and lipoprotein assembly in intestinal cells more than oleic acid.

To study the effect of OEA on liver derived cells, these cells were exposed to OEA or no lipids for 16 h. OEA reduced secretion of triglycerides and apoB in Huh-7 and HepG2 cells without affecting the secretion of apoAI. Further, MTP expression and activity was reduced in these cells after OEA treatment compared to no OEA controls. Similarly, OEA reduced MTP expression and activity as well as secretion of triglycerides in mouse primary hepatocytes. These studies indicate that OEA reduces MTP expression, lipid synthesis and lipoproteins secretion in liver derived cells. Thus, OEA has differential effects on MTP expression and lipoprotein assembly and secretion in intestine and liver derived cells. It is likely that OEA regulates postprandial response to optimize exogenous lipid absorption and reduce endogenous lipid mobilization.
Abstract 394: Bacterial Serum Opacity Factor Rescues HDL Functionality in SR-B1
-/-
Mice
Authors: Rosales Corina , Yelamanchili Dedipya , Gillard Baiba K , Gotto Antonio M , Pownall Henry J
Published: March 2019

Although plasma HDL-C levels negatively correlate with atherosclerotic cardiovascular disease (ACVD), attempts to reduce ACVD risk by raising plasma HDL have disappointed. Thus, hypotheses about salutary HDL effects have shifted from higher-is-better to function-is-more-important. The SR-B1
-/-
mouse is an extreme model of HDL dsyfunctionality; compared to WT mice, SR-B1
-/-
mice have higher plasma HDL levels and an HDL surface that is free cholesterol (FC)-rich (60 vs. 15 mol%). This would be expected to increase HDL-FC bioavailability to cytotoxic levels. HDL dysfunctionality among SR-B1
-/-
mice is associated with multiple metabolic abnormalities—impaired cell membrane structure and function and atherosusceptibility, despite having high plasma HDL-C levels; moreover, female SR-B1
-/-
mice are infertile. Liver-specific SR-B1 expression in SR-B1
-/-
mice normalizes HDL size and FC content. Thus, the SR-B1
-/-
mouse phenotype is due to lack of hepatic clearance of lipids from dysfunctional HDL. Serum opacity factor (SOF) is a bacterial protein that catalyzes the quantitative disproportionation of HDL into a cholesteryl ester-rich micro emulsion (CERM), neo HDL, and lipid-free apo AI. The CERM contains apo E and all HDL-CE. Injection of SOF (4 μg) into WT mice lowers plasma cholesterol by diverting the CERM to hepatic LDLR. Thus, we began testing whether adeno-viral delivery of SOF (AAV
SOF
) to SR-B1
-/-
mice rescues HDL functionality. A plasmid encoding the SOF gene was synthesized; SOF DNA was isolated by restriction enzyme digestion and cloned into a pAAV-TBG-mcs plasmid that was submitted to the PENN Vector Lab for virus production and isolation. AAV
GFP
plasmid (UPENN) was used as control. Good SOF production and secretion was confirmed by transfection of Huh7 hepatocytes. AAV
SOF
injection into SR-B1
-/-
mice induced constitutive plasma SOF activity and reduced HDL-C levels to nil. Superposition of high plasma HDL levels and a high mol% FC in SR-B1
-/-
is expected to increase HDL-FC bioavailablity that contributes to whole-body FC-toxicity and the observed metabolic abnormalities. Future tests will determine whether ablation of dysfunctional HDL in SR-B1
-/-
mice rescues their pathological phenotype, especially atherosclerosis.
Abstract 395: Changes in the HDL Lipidome With Regular Exercise: a Pilot Study
Authors: Sarzynski Mark A , Barupal Dinesh K , Showalter Megan R , Barber Jacob L , Ruiz-Ramie Jonathan J , Bouchard Claude , Fiehn Oliver
Published: March 2019

Background:
Over 200 individual molecular lipid species have been found to reside on high-density lipoproteins (HDL). The effects of exercise on the HDL lipidome profile is unknown.

Methods:
We examined changes in the HDL lipidome after regular exercise in 20 individuals who completed a 20-week endurance exercise program as part of the HERITAGE Family Study. The 20 subjects were selected based on discordance for exercise-induced increases in mean HDL particle (HDL-P) size (10 high-responders, 10 non-responders). The abundance of 184 known individual lipid species was quantified in baseline and post-training HDL plasma samples by UPLC-QTOF-MS. HDL was isolated through gel filtration chromatography.

Results:
Seven lipid species, particularly classes of triacylglycerols, cholesteryl esters, phosphatidylcholines, and sphingomyelins, showed nominal (p<0.05) increases in abundance with exercise training, with fold changes ranging from 1.13 to 1.65 (
Figure 1
). Three lipids showed differential responses to exercise between high- and non-responders of HDL-P size, including one lipid and metabolite each that showed an overall effect of training (
Figure 1
). For example, the abundance of sphingomyelin d39:2 decreased by an average of 23% in the high-responders of HDL size to exercise, whereas abundance increased by 71% in the non-responders (p=0.004 for difference between groups). None of the exercise-induced changes in lipid levels were statistically significant after accounting for multiple testing.

Discussion:
This pilot study showed limited evidence of systematic remodeling of the HDL lipidome in response to regular endurance exercise. Future studies are needed with larger sample sizes, diverse populations, and differing exercise protocols to further examine how the molecular composition of HDL particles changes in response to exercise and potentially contributes to its atheroprotective functions.
Abstract 409: Ambient Fine Particulate Matter Impairs Endothelial Progenitor Cells Through Card9-mediated Innate Immune Response
Authors: Cui Yuqi , Liu Xuanyou , Chen Zhiheng , Xu Huifang , Hao Hong , Hao Hong , Zhu Qingyi , Jiang Meng , Lin Xin , Sun Qinghua , Cui Lianqun , Liu Zhenguo
Published: March 2019

Background/Aims:
Endothelial progenitor cells (EPCs) play a critical role in angiogenesis and vascular repair. Some environmental insults, like fine particulate matter (PM) exposure, significantly impair EPCs through inflammation and ROS production. Cytosolic adaptor caspase recruitment domain 9 (CARD9) is important to the function of macrophages and involved in the innate immune response. The present study was to determine the role of macrophage-mediated immune response in PM-induced adverse effects on EPCs.

Methods:
PM was intranasal-distilled into male C57BL/6 mice for 1, 2, 7, 14 and 28 days. The level of EPCs (CD34
+
/Flk-1
+
, Sca-1
+
/Flk-1
+
, c-Kit
+
/CD31
+
and CD34
+
/CD133
+
) in the bone marrow (BM) and circulation was determined using flow cytometry along with the proliferation and apoptotic level of EPCs as well as intracellular reactive oxygen species (ROS) formation. Serum TNF-α, IL-1β, IL-6 and IL-10 were measured using ELISA. To determine the role of CARD9-mediated signaling in PM-induced detrimental effects on EPCs, CARD9 knock out (CARD9
-/-
) mice were used to repeat the experiment. The population of total macrophages as well as M1 and M2 subpopulations in both BM and blood were analyzed in the mice with and without PM exposure. To study the function of macrophages after PM exposure, the peritoneal macrophages were isolated and cultured for 48 hours to measure the production of TNF- α, IL-1β, IL-6 and IL-10 in the media.

Results:
PM exposure significantly decreased BM and circulating CD34
+
/FLk-1
+
as well as BM Scal-1
+
/Flk-1
+
population, promoted apoptosis and inhibited proliferation of EPCs in association with increased ROS production, serum TNF-α, IL-1β, IL-6 and decreased serum IL-10 level. These PM-induced effects on EPC population and cytokine production were largely prevented in CARD9
-/-
mice along with increased M2 population in both BM and blood. The production of TNF-α, IL-1β and IL-6 was significantly decreased, while the IL-10 level increased, from cultured macrophage of CARD9
-/-
mice compared with WT control.

Conclusion:
PM exposure significantly decreased BM and circulating EPCs population due to CARD9-mediated innate immune response.
Abstract 413: CD11c/CD18 Affinity Modulates Monocyte Inflammatory in Primary and Recurrent Myocardial Infarction
Authors: Hernandez Alfredo A , Foster Greg , Fernandez Andrea , Simon Scott , Singh Gagan
Published: March 2019

Several studies address the role of CD11c
+
monocytes in the exacerbation of atherosclerosis through the maintenance of macrophages, notably foam cells, however this cell type remains elusive in the context of human atherosclerosis. Intermediate monocytes (CD14+CD16+, Mon2) have a high degree of potential participation in atherosclerosis through the upregulation of membrane β2-integrin CD11c/CD18 expression that functionally orchestrates their recruitment to VCAM-1 activating VLA-4 leading to focal clustering and shear resistant arrest. Both Mon2 frequency and CD11c expression remain critical factors associated with the extent of coronary artery disease (CAD) related to persistent inflammation, plaque growth and destabilization, myocardial infarction and morbidity. Understanding dynamic alteration of Mon2 CD11c and its function in promoting macrophage accumulation in plaques that exacerbate and progress atherosclerotic disease progression toward myocardial infarction remains undetermined. In this study, we investigated the activation status of circulating Mon2 in patients experiencing non-ST myocardial infarction (NSTEMI) compared to those undergoing angiography in the cardiac clinic. Mon2 activation status and CD11c function was gauged using flow cytometry and real-time microfluidics using an
Artery-on-a-Chip
(A-Chip) device. Circulating Mon2 upregulated CD11c receptors to the greatest extent in patients experiencing myocardial infarct that increase with recurrence and lead to enhanced CD11c dependent capture on VCAM-1 relative to patients being treated in the cardiac clinic. Arrest on VCAM-1 under shear flow resulted in time-dependent phenotypic alteration including the loss of CD16 over 45 min and was dependent on atherosclerotic disease but independent of NSTEMI and involved CD11c outside-in signaling. A shift from a high to low affinity state of CD11c triggered the translocation of several activation and differentiation markers such as NfKB, and upregulation of MMP-9 associated with an M1 phenotype. CD11c receptor number and subsequent signaling provides a potential target for intervention of atherogenesis by altering Mon2 differentiation potential and inflammatory capacity.
Abstract 424: Waist Circumference for the Diagnosis of Metabolic Syndrome on Risks of Arterial Stiffness and Diabetes
Authors: Tsai Sung-Sheng , Chu Pao Hsien
Published: March 2019

Background:
Published studies seldom tested the weight of different waist circumference (WC) cut-off values for the diagnosis of metabolic syndrome (MetS) in predicting clinical outcomes, including cardiovascular disease and diabetes.

Methods:
This is a Chinese population-based cross-sectional study screening subjects from a Health Examination Program since 1999 to 2015. The MetS identification and scores were determined either according to the Adult Treatment Panel III/ American Heart Association/National Heart, Lung, and Blood Institute (ATP III/AHA/NHLBI)- or Asian-WC cut-off points. The developments of a higher brachial-ankle pulse wave velocity (baPWV) and diabetic-level hyperglycemia were surveyed by comparing the areas under receiver operating characteristic curves (AUC-ROC) for both MetS scores.

Results:
According to the ATP III/AHA/NHLBI- or Asian-MetS criteria, 6633 (24.8%) or 9133 (34.2%) subjects were diagnosed as the MetS among 26735 study subjects with a mean age of 55 ± 12 years. The stepwise increases in baPWV and prevalence of diabetic-level hyperglycemia were associated with both MetS scores after adjusting for age and sex. The AUC-ROC for the ATP III/AHA/HLBI- vs Asian-MetS scores to predict a higher baPWV (0.685 vs 0.680,
p
=0.271) and diabetic-level hyperglycemia (0.791 vs 0.784,
p
=0.546) were similar.

Conclusions:
In a stepwise manner, both ATP III/AHA/NHLBI- or Asian-MetS scores were strong risk factors for arterial stiffness and diabetes. Through a novel and holistic approach, the predictive performance of the ATP III/AHA/NHLBI-MetS score for the risks of arterial stiffness and diabetes was comparable to the Asian-MetS score among a Chinese population. 
Abstract 427: Chronic Intermittent Hypoxia Induces Endothelial Insulin Resistance
in vivo
and
in vitro
Authors: Li Linyi , Yang Yunyun , Ma Xinliang , Wei Yongxiang , Qin Yanwen
Published: March 2019

Introduction:
Obstructive sleep apnea, a condition leading to chronic intermittent hypoxia (CIH), is an independent risk factor for cardiovascular disease and type 2 diabetes and is correlated with insulin resistance. Insulin stimulates production of nitric oxide (NO) in vascular endothelial cells through the IRS-1/PI3K/Akt/eNOS pathway (IRS-1, insulin receptor substrate 1; PI3K, phosphatidylinositol 3-kinase; eNOS, endothelial NO synthase). Endothelial cells directly expose to blood flow and respond to oxygen concentration change in flow. We wondered if and how CIH would affect insulin signalling/action in endothelial cells.

Methods and Results:
C57BL/6J mice were exposed to CIH (21%-5%, 90 s/cycle, 10 h/day) or intermittent air for 7 weeks. HUVECs were exposed to CIH (21%-5%, 1 h/cycle) for 24 h or 48 h with or without insulin. We observed that CIH increased systolic blood pressure and impaired endothelium-dependent relaxation of aortas in C57BL/6J mice. CIH increased IRS-1 phosphorylation at Ser307 and Ser612 and impaired insulin-stimulated phosphorylation of IRS-1 at Tyr896 and Akt/eNOS pathway in aortas. Furthermore, CIH activated hypoxia-inducible factor 1α (HIF-1α) and nuclear factor kappa B (NF-κB) in aortas. In vitro, CIH activated endothelial HIF-1α signaling, impaired endothelial insulin signaling and reduced insulin-mediated NO production. In addition, CIH resulted in mitochondria proton leak and elevated oxygen consumption in HUVECs. Increased oxygen consumption further leaded to endothelial hypoxia and HIF-1α induction. Pharmacological inhibition of proton leak ameliorated CIH-induced endothelial insulin resistance in HUVECs.

Conclusion:
Our data suggest that CIH induces insulin resistant endothelial dysfunction, which may be associated with HIF-1α induction and proton leak.