bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 202: Enrichment of Transcription Factor Binding Sites in the Promoter Regions of Differentially Expressed Genes in Vascular Smooth Muscle Cells from Abdominal Aortic Aneuryms.
Authors: Airhart Nathan , Curci John 
Published: July 2021

Background
We have previously shown that VSMC from AAA are unique compared to cells from normal aorta (NAA) and carotid endarterectomy (CEA) with increased production of matrix metalloproteinases and elastin degrading activity. The purpose of this study was to explore the mechanisms behind this phenotype.

Methods
Tissue for VSMC cultures was obtained from patients undergoing AAA repair and CEA. NAA tissue was obtained from renal transplant patients (NAA). Total RNA was isolated from VSMC and subjected to whole-genome microarray. Enrichment of binding sites for transcription factors (TF) within 5 kD of transcription start sites of upregulated genes were identified using Whole Genome rVista. Enriched gene ontology terms were identified using the Database for Annotation, Visualization, and Integrated Discovery (DAVID).

Results
Gene profiles of 22 AAA, 29 CEA, and 17 NAA cell lines were compared. We identified 120 upregulated genes in AAA-VSMC relative to NAA and CEA-VSMC (FDR<0.05). Analysis of transcription factor binding sites of these genes showed enrichment of TFs including members of the ETS, AP-1, and Rel/Ankyrin families. Gene ontology (GO) revealed enrichment of developmental process and immune system genes. Analysis by cell compartment showed enrichment of extracellular matrix and intermediate filament cytoskeleton genes (Table 1).

Conclusion
This is the first study to demonstrates enrichment of TF families such as ETS, AP-1 and Rel-Ankyrin in AAA VSMC. This suggests that VSMC in AAA may not just be responding to inflammatory or other local stimuli, but may be directly contributing to the ECM changes that define AAA.
Abstract 230: Effect of aerobic exercise on Endothelial Progenitor Cells (EPCs) in patients with prediabetes
Authors: Sen Sabyasachi , Chou Cyril , Pollard Nicole , Islam Ashequl 
Published: July 2021

Introduction
At least 79 million Americans have pre-diabetes. Though life-style modification prevents progression from Pre-diabetes to overt diabetes, the effect on vascular endothelial progenitor cells (EPC) stem cells is unknown.

Methods
This is a crossover study of 16-week duration, using exercise-naive pre-diabetes patients, n=12, aged 45-65 yrs with a BMI of 25-34.9. We studied flow mediated dilatation (FMD), EPC function and gene expression and serum endothelial inflammatory levels after 6 weeks each of aerobic exercise (150min/week) and non exercise phase, in a cross over design with 4 week wash-out period between the 2 phases. EPC migration was noted in response to chemotactic factors such as Vascular Endothelial Growth Factor (VEGF-A, 0- 50ng/ml) and Stromal Cell-Derived Factor-1(SDF1α, 0-100 ng/ml). Exercise compliance was monitored by regular phone calls and downloadable accelerometers.

Results
FMD studies (undertaken by 3 observers) showed mean FMD in non-exercise group of 5.7+0.6% which improved to 11.2+0.9% post-exercise. No significant weight loss noted between the 2 groups, however, significant reduction in leptin, IL-6, TNFα, hs-CRP, Triglyceride, and ApoB levels was noted. EPC gene expression analysis showed decrease of eNOS (5 fold) and increase of PECAM-1 (4-fold), Endothelin-1 (3-fold), IL-6 (2-fold), TNFα (3-fold) in non-exercise which improved following exercise. EPC migration improved post exercise, particularly in response to SDF-1α as did CD34+ cell number and EPC colony formation improved significantly also.

Conclusion
We demonstrate that pre-diabetic state is associated with poor vascular reactivity and impaired EPC stem cell function. Pre-diabetes may be a clinical window of therapeutic intervention opportunity when short course of aerobic exercise may allow improvement of vascular reactivity and EPC function despite no statistically significant weight loss. This study also indicates EPCs may act as an important biomarker for vascular function.
Abstract 231: Bone Marrow-Derived Mesenchymal Stem Cells from Amputated Limbs of Patients with Critical Limb Ischemia Provide Stromal Support for Endothelial Cells
Authors: Wang Ruoya , Copland Ian , Li Haiyan , Peister Alexandra , McDevitt Todd , Brewster Luke 
Published: July 2021

Critical limb ischemia (CLI) often results in amputation, but autologous cellular therapy using bone marrow-derived mesenchymal stem cells (MSCs) may be able to prevent this outcome. However, the quality of MSCs in the CLI population is not clear. Amputated CLI limbs contain reservoirs of MSCs that can be used to characterize their stromal capacity. We hypothesize that MSC stromal capacity is relatively preserved in ischemic limbs compared to ones from healthy donors.

Methods
Healthy donor MSCs (hdMSCs, n=2) and ischemic MSCs (iMSCs, n=3) from amputated limbs of non-diabetic CLI patients were cultured. Human microdermal ECs were commercially obtained. Co-culture pellets (1:1 ratio) of hdMSC:EC, iMSC:EC, and EC alone were formed and cultured in 3-D fibrin hydrogels in EC growth media. Images of the pellets and invasion areas were captured up to 5 days. The invasion area was quantified as the difference between the total and pellet area normalized to initial pellet area. Experiments were performed in quadruplicates and co-culture experiments were duplicated.

Results
iMSC:EC co-culture provided similar stromal support as the healthy donor cohort. Both MSC groups were significantly greater than the ECs alone following day 2 (see figure, *P<0.0005, ANOVA). Both MSC groups also exhibited significant increase in invasion area with time (P<0.01) while the ECs did not (P>0.5).

Conclusion
This is the first study to demonstrate that iMSCs provide similar stromal support to ECs as healthy hdMSCs, thus iMSCs may be of sufficient quality for use in autologous therapy. We are currently quantifying secretome expressions of the iMSCs to provide insights into mechanisms of the stromal support.
Abstract 27: Dok-1 Negatively Regulates Integrin aIIbß3 Outside-in Signaling and Inhibits Thrombosis
Authors: Niki Masaru , Jin Hong , Chauhan Anil K , Lentz Steven R 
Published: July 2021

Background
Platelet agonists activate integrin αIIbβ3 to allow the binding of soluble fibrinogen (a process known as inside-out signaling). Subsequent platelet aggregation leads to αIIbβ3 outside-in signaling, which results in tyrosine phosphorylation of β3 and other proteins, cytoskeletal reorganization, and platelet spreading. It has been reported that the adapter protein Dok-1 binds to the cytoplasmic tail of β3 and inhibits αIIbβ3 activation, but the specific role of Dok-1 in regulating inside-out or outside-in platelet signaling remains undefined.

Methods

We assessed the effects of Dok-1 on platelet signaling and thrombosis in Dok-1 null (Dok-1-/-) mice. Inside-out signaling was assessed by measuring αIIbβ3 activation (using the JON/A antibody) and fibrinogen binding by flow cytometry after stimulation of platelets with thrombin, ADP, and/or the thromboxane A2 receptor agonist
U46619.
Outside-in signaling was examined by measuring platelet spreading and clot retraction. Tail-transection bleeding time and susceptibility to thrombotic occlusion of the carotid artery in response to photochemical injury (rose bengal) were also measured.

Results
No significant differences in JON/A or fibrinogen binding were detected between wild-type (Dok-1+/+) and Dok-1-/- platelets, suggesting that Dok-1 does not regulate inside-out signaling. In contrast, Dok-1-/- platelets exhibited increased clot retraction and enhanced spreading on fibrinogen upon thrombin stimulation compared to Dok-1+/+ platelets (P<0.05), suggesting that Dok-1 negatively regulates outside-in signaling. Compared with Dok-1+/+ mice, Dok-1-/- mice had shorter bleeding times (181±168 vs. 379±193 seconds; P<0.001) and Dok-1-/- mice had shorter times to stable occlusion times of the carotid artery after photochemical injury compared with Dok-1+/+ mice (16.4±6.1 vs. 25.0±8.1 minutes; P<0.05).

Conclusions
The adaptor protein Dok-1 functions to negatively regulate integrin αIIbβ3 outside-in signaling. Deficiency of Dok-1 results in a prothrombotic phenotype, with shorten bleeding times and enhanced arterial thrombosis.
Abstract 280: Inhaled Carbon Monoxide Attenuates Aortic Aneurysmal Growth in a Topical Elastase Mouse Model
Authors: Shukla Ankur J , Washington Chris , McEnaney Ryan , Tzeng Edith 
Published: July 2021

Objective
Abdominal aortic aneurysms (AAA) affect up to 9% of men over 65 years of age with increasing incidence with further aging. Little is still known about the pathogenesis of AAA but inflammation appears to be one of the main contributing factors. Carbon monoxide (CO) is an endogenously produced molecule that has been shown to have significant anti-inflammatory and cytoprotective properties. Thus, we hypothesize that exogenous CO may attenuate AAA growth.

Methods
Eight week-old C57Bl6 mice were anesthetized and infrarenal aorta were exposed, measured in 3 locations, and treated with topical porcine pancreatic elastase (30 ul). Mice were either maintained in room air for the duration of the experiment or treated with inhaled CO (250 PPM) for 1 hr daily beginning on day 1. Animals were sacrificed on day 14 when aortic diameters were measured and tissues collected for morphometric analysis.

Results

Topical elastase treatment resulted in marked aortic enlargement with true aneurysm formation (253% of baseline, N=8). However, CO treatment reduced aneurysm size significantly (222%, N=8; p-0.039) with noticeable reduction in adhesion formation and scarring around the aorta. Histologically, CO treated animals maintained a larger medial area than controls (0.092 mm
2
vs 0.057 mm
2
; p = 0.023) (Figure 1).

Conclusion
Inhaled CO appears to mitigate aneurysmal degeneration of the abdominal aorta induced by topical elastase. There was a clear reduction in adhesions to the aorta, strongly supporting the anti-inflammatory mechanism involved. The maintenance of a thicker medial component of the aorta in CO treated mice also suggests protection of the smooth muscle cells.
Abstract 301: Angiotensin II-induced Aortic Stiffening is Associated With Upregulation of Mir-21 And Downregulation of Mir-29b
Authors: Raaz Uwe , Toh Ryuji , Adam Matti , Nakagami Futoshi , Maegdefessel Lars , Deng Alicia , Spin Joshua M , Tsao Philip S 
Published: July 2021

Background
Arterial stiffening is associated with aging, as well as numerous diseases such as hypertension, congestive heart failure, diabetes mellitus and renal failure, and has been identified as an independent risk factor for cardiovascular disease. Consequently there is a growing interest in the mechanisms that govern the process of arterial remodeling. Recently, small noncoding RNAs, termed microRNAs (miRs), have emerged as powerful cellular regulators involved in disease and tissue remodeling. More specifically, miR-29b downregulation as well as miR-21 upregulation have been identified as pro-fibrotic mechanisms in various cardiovascular disease models. This study investigated the role of miR-21 and miR-29b in a mouse model of AngII-induced arterial remodeling.

Materials and methods
Angiotensin II (AngII) (1000ng/kg/min) was infused via osmotic pumps in 10-week-old apoE-/- male mice (C57BL/6J background) for 7 days. Subsequently, arterial stiffness was determined in vivo using ultrasound-based measurements. Abdominal aortas were harvested and expression of various collagen isoforms (Col1a1, Col3a1, Col5a1) known to be crucial determinants of arterial remodeling/stiffening was quantified via qRT-PCR. We also measured expression levels of miR-21 and miR-29b.

Results
AngII stimulation resulted in an increased aortic stiffness paralleled by a marked pro-fibrotic response as evidenced by significant increases in Col1a1, Col3a1, and Col5a1 in the infrarenal aorta compared to baseline levels (p < 0.05). This increase was accompanied by significant downregulation of miR-29b, and upregulation of miR-21 (p <0.05).

Conclusion
The pro-fibrotic response in AngII-mediated arterial remodeling is associated with an increase in miR-21 and a decrease in miR-29b. Modulation of miR-21 and miR-29b have both been successful in altering fibrotic mechanisms in various cardiovascular diseases. These data suggest they may also be potential targets in the treatment of hypertensive vascular remodeling/arterial stiffening.
Abstract 309: Osteogenic Induction of p-ERK 1/2 Signaling Associated with NOTCH1 Inactivation in Human Aortic Valvar Interstitial Cells
Authors: Godby Richard C , Munjal Charu , Opoka Amy M , Narmoneva Daria A , Hinton Robert B 
Published: July 2021

Introduction

Heart valve disease affects over 2% of the population, leading to at least 20,000 deaths and $1 billion in direct costs in the US each year. Calcific aortic valve disease (CAVD) is linked to loss of function mutations in
NOTCH1
that result in osteogenic gene induction.
In vitro
animal studies have established that multiple osteogenic targets are mediated through induction of p-ERK 1/2 signaling; however, the relationship between NOTCH1 and p-ERK has not been examined in primary human aortic valvar interstitial cells (AVICs).

Hypothesis
We tested the hypothesis that chemically-induced NOTCH1 suppression results in mineralization due to p-ERK 1/2 activation in human AVICs.

Methods
Human AVICs were isolated from CAVD specimens at the time of valve replacement. AVICs were cultured for two weeks in complete media containing DMEM and 10% FBS, and treated with calcific media (10mM b-glycerol phosphate; 60μM ascorbic acid-2-phosphate; 1μM dexamethasone), which has been shown to suppress NOTCH1 signaling. Standard real-time RT-PCR, western blot, and immunocytochemistry were used to compare groups.

Results

Treatment with calcific media resulted in a 2.3 fold decrease in the level of
NOTCH1
expression and a 3 fold increase in p-ERK 1/2 expression, assessed with both western blots and immunocytochemistry. Interestingly, calcific media also induced a 3.5 fold decrease in VEGF, consistent with early osteogenic differentiation. Additionally, the AVICs treated with calcific media demonstrated overt calcification by Alizarin Red staining.

Conclusions

Human AVICs demonstrated calcification associated with p-ERK 1/2 activation and reduced
NOTCH1
levels, suggesting p-ERK inhibition may mitigate the effects of NOTCH loss of function, thereby representing a potential pharmacologic therapy for CAVD. Further investigation of the molecular mechanisms linking NOTCH1 and p-ERK are warranted, including studies that specifically inhibit NOTCH1 signaling and define the impact of mechanical forces on molecular pathways, thereby identifying mechanisms underlying CAVD progression.
Abstract 311: TGFb/Smad3 Stimulation Drives Smooth Muscle Cell De-differentiation
Authors: DiRenzo Daniel M , Shi Xudong , Seedial Stephen , Guo Lian-wang , Liu Bo , Kent K. Craig 
Published: July 2021

Restenosis, or vessel re-narrowing, occurs in approximately 25-50% of arterial interventions involving balloon angioplasty due to the formation of a proliferative plaque in the vessel lumen termed neo-intimal hyperplasia. Arterial smooth muscle cells (SMCs) contribute to neo-intimal hyperplasia through a de-differentiation process that includes downregulation of their contractile gene expression and conversion to a phenotype that includes proliferation, migration, and matrix synthesis. Expression of TGFβ and its downstream signaling protein, Smad3, are greatly upregulated following vascular injury, including balloon angioplasty. Classically, TGFβ signaling has been shown to suppress SMC proliferation and migration in vitro, however, Smad3 overexpressing SMCs demonstrate enhanced proliferation and migration. Furthermore, overexpression of Smad3 in rat carotid arteries enhances neo-intimal hyperplasia following balloon angioplasty. These results lead us to hypothesize that TGFβ signaling, in the context of upregulated Smad3, drives SMC de-differentiation leading to enhanced cellular proliferation and migration.
We utilized primary rat SMCs infected with adenovirus constructs overexpressing Smad3 or GFP control and performed gene expression microarrays 24 hours following TGFβ administration. We observed statistically significant (p<0.05) upregulation of 145 genes and downregulation of 76 genes by more than 3-fold. GO term analysis revealed that genes involved in embryonic tissue development (41 genes) and stem/progenitor cell differentiation (27 genes) were significantly enriched in TGFβ/Smad3 stimulated cells. Confirmatory qRT-PCR demonstrated that the contractile genes SM-MHC, smooth muscle actin, and calponin were significantly downregulated -6.3, -2.7 and -2.1 fold, respectively. In contrast, stem/developmental related genes Cxcr4, Cd34, Wnt11, Wnt2b and IL11 were significantly upregulated by 105.2, 22.3, 11.5, 14.0, and 12.5 fold, respectively. These results strongly suggest that TGFβ/Smad3 stimulation is a powerful de-differentiation signal in SMCs and plays an important role in the development of neo-intimal hyperplasia.
Abstract 320: Heterogeneous Coronary Stent Inflammation and Healing Revealed by in vivo Optical Molecular-Structural Imaging
Authors: Osborn Eric A , Hamidi Ehsan , El Maghraby Amr , Yoo Hongki , Mauskapf Adam , Tearney Guillermo J , Jaffer Farouc A 
Published: July 2021

Introduction

Patients with diabetes mellitus that receive coronary stents experience greater stent restenosis and thrombosis, leading to adverse clinical outcomes. Impaired stent healing is linked to elevated endovascular inflammation, but
in vivo
data is lacking. Here, we assessed stent inflammation and tissue healing in diabetic rabbits, using intravascular molecular-structural near-infrared fluorescence (NIRF)-optical frequency domain imaging (OFDI).

Methods

A bare-metal stent (3.5x12mm) was implanted in the infrarenal aorta of alloxan-induced diabetic rabbits (n=5). At day 28, intravascular NIRF-OFDI was performed. Prosense VM110 (5mg/kg IV 24 hrs before imaging; ex/em 750/780 nm) enabled NIRF molecular imaging of inflammatory cathepsin activity. Structural OFDI stent endothelialization (coverage) was simultaneously assessed in 0.5mm intervals. Aortas were then harvested for
ex vivo
fluorescence reflectance imaging (FRI), electrochemical stent dissolution, and cathepsin B immunostaining.

Results

At day 28, stent NIRF inflammatory cathepsin protease activity was enhanced compared to the non-stented aorta (50.9±2.0 vs. 26.1±0.6 nM; p<0.0001). OFDI demonstrated greater average stent edge restenosis (proximal and distal 2mm stent edge neointimal area, 1.63±0.12 vs. mid 2 mm stent 0.50±0.05 mm
2
; p<0.0001), with less de-endothelialized struts at the stent edges (2.1±1.7% vs. 14.0±4.1% mid stent; p=0.03). Stent edge NIRF inflammation was also greater (60.8±3.4 vs. 40.8±1.4 nM mid stent; p=0.0005), and correlated with neointima formation (R=0.43; p=0.003). Conversely, NIRF inflammation negatively tracked with OFDI-uncovered stent struts (R=-0.42; p=0.004). FRI corroborated the enhanced stent-edge NIRF pattern. Matched histopathology revealed cathepsin B expression in NIRF-positive regions.

Conclusions
Intravascular NIRF-OFDI molecular-structural imaging demonstrates that NIRF inflammatory protease activity is linked to increased bare-metal stent edge neointimal formation, but inversely relates to uncovered stent struts. This translatable approach provides new insights into stent inflammation and healing, and may ultimately inform the risk of stent restenosis and thrombosis.
Abstract 323: Increased Levels of Plasma Soluble Sema4D in Patients with Heart Failure
Authors: Lu Qiongyu , Dong Ningzheng , Yi Wenxiu , Wang Yuxin , Zhang Shengjie , Tang Xiaorong , Wu Qingyu , Brass Lawrence , Zhu Li 
Published: July 2021

Introduction
Semaphorin 4D (Sema4D/CD100) is a 150-kDa transmembrane glycoprotein expressed by platelets and T-cells. When these cells are activated, Sema4D is cleaved proteolytically, generating a biologically active 120-kDa fragment (soluble Sema4D) capable of targeting receptors on platelets, B-cells, endothelial cells and tumor cells. However, its plasma levels and significance in cardiovascular disorders have not been reported.

Hypothesis
We hypothesized that plasma soluble Sema4D levels increases in patients with heart failure (HF) and that such an increase reflects T-cell and/or platelet activation.

Methods
We established an ELISA method and detected soluble Sema4D levels in human plasma, and evaluated the effect of estradiol on Sema4D shedding in platelets with collagen-induced aggregation. Percentage of Sema4Dhigh T cells, B cells and platelets in HF patients and healthy controls was detected with flow cytometry.

Results
In healthy controls, plasma Sema4D levels were higher in men than women (5.15 ± 3.30, n=63, vs. 4.19 ± 2.39 ng/mL, n=63, P<0.05), and cleavage of Sema4D in platelets activated by collagen was inhibited by estradiol. In HF patients, plasma Sema4D levels were significantly higher than those in healthy controls (8.94 ± 5.89 ng/mL, n=157 vs. 4.67 ± 2.99 ng/mL, n=126, P<0.0001) with the highest levels being in HF patients with diabetes mellitus (DM) (10.45 ± 5.76 ng/mL, n=40). We also found that there was a higher percentage of Sema4Dhigh CD3+ (P<0.01), CD4+ (P<0.001), and CD8+ (P<0.01) T-cells in samples from HF patients, but no change in B cells and platelet Sema4D expression levels.

Conclusions
Plasma Sema4D levels are increased in HF patients, especially in those who also have diabetes. There was an accompanying increase in the Sema4Dhigh population of T-cells, suggesting a potential role of these T-cells in heart failure.