bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 212: MicroRNA-663 is a Novel Regulator of Human Vascular Smooth Muscle Cell Phenotypic Switch in vitro and Neointimal Formation in vivo
Authors: Li Pan , Yi Bing , Qin Qing , Chen Ming , You Xiaohua , Zhao Xianxian , Qin Yongwen , Sun Jianxin 
Published: July 2021

Background
Abnormal phenotypic switch of vascular smooth muscle cell (VSMC) is a hallmark of vascular disorders such as atherosclerosis and restenosis after angioplasty. Recently, microRNAs (miRNAs) emerge as critical regulators for vascular smooth muscle cell (VSMC) function. Our initial study identified miR-663 as one of the most sharply downregulated miRNAs in human proliferative aortic smooth muscle cells.

Hypothesis
MiR-663 is implicated in human VSMC phenotypic switch and the development of neointima formation.

Methods and Results
By using quantitative real-time PCR (qRT-PCR), we found that microRNA-663 (miR-663) was significantly downregulated in cultured human aortic VSMCs upon platelet-derived growth factor (PDGF) treatment, whereas its expression was markedly increased during VSMC differentiation as induced by either retinoid acid or SMC differentiation medium, a condition which induces SMC differentiation and inhibits cell proliferation. Furthermore, we demonstrated that overexpression of miR-663 significantly increased the expression of VSMC differentiation marker genes, such as SM22α, SM α-action, calponin, and SM myosin heavy chain, suggesting that miR-663 is a novel modulator implicated in human VSMC phenotypic switch. Moreover, miR-663 potently inhibited PDGF induced VSMC proliferation and migration. Mechanistically, we identified JunB as a downstream target of miR-663 in human VSMCs. Indeed, overexpression of miR-663 markedly inhibited the expression of the transcription factor JunB as well as its downstream molecules including matrix metallopeptidase-9 (MMP-9) and myosin light chain-9 (Myl9), thus inhibiting VSMC proliferation and migration. Finally, we showed that adeno-miR-663 markedly suppressed the neointimal lesion formation by approximately 50% in mice after vascular injury induced by carotid artery ligation, specifically via decreased JunB expression.

Conclusion
These results indicate that miR-663 is a novel modulator implicated in human VSMC phenotypic switch through targeting JunB expression and suggest that specific modulation of miR-663 in human VSMCs may represent a novel and attractive approach for the treatment of vascular proliferative diseases.
Abstract 214: Protein Restriction Attenuates Intimal Hyperplasia and Alters Blood Lipid Profiles
Authors: Mauro Christine R , Tao Ming , Yu Peng , Liu Chengwei , Mitchell James , Zheng Hanqiao , Kristal Bruce S , Bird Susan S , Ozaki C. Keith 
Published: July 2021

Introduction
Dietary restriction (DR: reduced enteral intake without malnutrition) diminishes the acute phase response to surgical stress in preclinical models. We hypothesized that short-term preoperative protein-free DR would attenuate the vascular response to injury (intimal hyperplasia: IH) while perturbing systemic circulating lipids.

Methods
After 2 weeks of 60% fat kcal diet, 8 week old B6D2F1/J mice had ad lib access to a complete 10% fat kcal diet (n=10) or a reduced calorie, protein-free diet (DR group, n=10) 1 week before induction of IH, then an ad lib complete diet. IH was induced via a validated model placing a nylon suture tie around the distal carotid artery and external 35g needle mandrel (outer diameter=0.14mm). Subsequent removal of the mandrel created a focal area of stenosis (~78% lumen diameter/~85% flow reduction). 4 weeks later, tissues were harvested for morphology and immunohistochemistry for CD45. Separately, serum was collected from mice fed a complete or DR diet for 1 week absent any surgical stress (n=20/group) for mass spectrometry-based lipidomics.

Results
DR mice showed less intimal area (p = 0.032) vs controls with statistically equivalent intimal leukocyte infiltration. DR mice also had significantly larger internal elastic lamina length (p = 0.003), a remodeling measure. DR serum exhibited significant decreases in certain classes of circulating lipids, including a collapse of multiple triglyceride types.

Conclusions
One week of protein-free DR dramatically decreased circulating lipids and attenuated arterial IH. Preoperative dietary manipulations may offer a practical means of extending durability of vascular interventions.
Abstract 215: Targeting MT1-MMP via Polymeric Transfection is More Efficient to Inhibit the Cellular Processes of Intimal Hyperplasia
Authors: Mountain Deidra J , Arnold Joshua D , Kirkpatrick Stacy S , Stevens Scott L , Goldman Mitchell H , Freeman Michael B , Grandas Oscar H 
Published: July 2021

OBJECTIVE
Polymeric transfection could be a valuable therapeutic tool as it may be less toxic and more predictable than traditional methods, which are often poorly translated to in vivo models. Matrix metalloproteinases (MMP), in particular MT1-MMP, play a role in intimal hyperplasia (IH) and their targeted inhibition should be examined in vivo for the prevention of IH. Here we established conditions for polymeric transfection in vascular cells and ex vivo tissue and assayed its efficiency in targeting MMP-modulated cellular processes of IH.

METHODS
Vascular smooth muscle cells (HASMC) and rodent carotid artery segments were transfected with poly(β-amino ester) polymers (Stem) or equivalent DharmaFECT2 (Dharm) conjugated to GAPDH, negative control (NC) or MT1-MMP siRNA.

RESULTS
Bioconjugate translocation was observed at 0.65μl polymer: 100pmol siRNA/ml (Fig 1A). Gene silencing was polymer concentration dependent in vitro, and the most efficient bioconjugate (1.95μl polymer: 100pmol siRNA/ml) resulted in significant silencing ex vivo (Fig 1B). Using these empirically derived conditions, polymeric silencing of MT1 resulted in increased gene knockdown (Fig 2A) and inhibition of HASMC migration (Fig 2B) and proliferation (Fig 2C) compared to Dharm.

CONCLUSIONS
HASMC and arterial tissue were successfully transfected, and silencing of MT1 via polymeric transfection was more effective in gene knockdown and in inhibiting the cellular processes of IH development in vitro when compared with traditional techniques. Future studies using targeted polymeric gene therapy to inhibit MT1 in animal models may modify vascular response to injury and prevent vascular disease.
Abstract 221: Integral Role of Akt-Serum Response Factor-Myocardin Signaling in Myofibroblast Differentiation with Implications in Pulmonary Arterial Hypertension
Authors: Abdalla Maha , Goc Anna , Segar Lakshman , Somanath Payaningal R 
Published: July 2021

Objectives

Myofibroblast (MF) differentiation is marked by the
de novo
expression of smooth muscle alpha-actin (αSMA) stress fibers, a function critical for tissue repair both physiologically and pathophysiologically. However, the role of Akt in regulating MFs with implications in pulmonary arterial hypertension (PAH) and fibrosis, and the effect of pharmacological inhibition of Akt pathway on PAH and fibrosis remains poorly defined.

Methods
We used NIH 3T3 fibroblasts and primary human lung fibroblasts (HLFs) in the current study. Cells were treated with TGFβ, the profibrotic cytokine and the role of Akt, SRF and myocardin in αSMA expression, MF differentiation and collagen gel contraction was studied with the use of various constructs encoding expression of specific genes.

Results
Here, we show that Akt mediates MF differentiation as evident by a 5-fold increase in αSMA expression and assembly in mouse embryonic fibroblasts (NIH 3T3) over-expressing active Akt1 (myr-Akt). As for the molecular mechanisms governing MFs, we found that αSMA expression was mediated by Akt, independent of mTOR; noteworthy, fibronectin expression, an ECM protein, is Akt-mTOR dependent. Inhibiting Akt activation utilizing triciribine blunted αSMA and its transcriptional activators, myocardin and SRF, inhibiting MF differentiation in NIH 3T3, myr-Akt-NIH 3T3, and human lung fibroblasts. Furthermore, blocking Akt reversed MF differentiation as evident by a blunted gel contraction primed with TGFβ.

Conclusions
In conclusion, our study clearly demonstrates the crucial role of Akt in mediating TGFβ-induced MF differentiation. We show a previously uncharacterized link between Akt and TGFβ-induced αSMA synthesis through myocardin and SRF, independent of mTOR, in MFs. These data constitute evidence of novel signaling cascade and targeting Akt pathway might be beneficial for the treatment of PAH and fibrosis.
Abstract 223: Is Oxidative Stress Important in AAA pathogenesis?
Authors: Urbonavicius Sigitas , Urbonaviciene Grazina , Sandermann Jes , Lindholt Jes S 
Published: July 2021

OBJECTIVE
Active investigations continue to identify markers other than size that would predict a risk of abdominal aortic aneurysms (AAA) rupture. Circulating biomarkers could also indicate optimal intervals between the surveillance intervals. Finally, the identification of biomarkers also may identify potential pathogenic pathways, and thus may open possibilities for pharmacological inhibition of growth. In the search of novel biomarkers of AAA progression, serum and wall material proteins were analyzed by a differential proteomic approach.

METHODS AND RESULTS
The same layers of AAA wall from ruptured (rAAA) and non-ruptured AAA were incubated, and the proteins released were analyzed by 2-dimensional difference in-gel electrophoresis. Proteins from serum were analyzed and correlated with AAA annual expansion rate. Several differentially expressed proteins involved in main AAA pathological mechanisms were identified by mass spectrometry. Among the 8 proteins identified in AAA wall, peroxiredoxin-2 (PRX-2) was more permanent, which was further validated by Western blot and immunohistochemistry. We demonstrated increased PRX-2 levels in rAAA patient wall material compared with AAA subjects and also positive correlation in serum among PRX-2 and AAA diameter and annual AAA expansion rate.

CONCLUSIONS
Eight proteins associated with AAA pathogenesis have been identified in AAA wall by a proteomic approach. Protein profiles identified in the wall from the rAAA and the serum would appear to be a convenient monitoring tool that has the ability to be both sensitive and specific for AAAs. Among them, PRX-2 serum levels are increased in AAA patients and correlate with AAA size and growth rate, suggesting the potential use of PRX-2 as a biomarker for AAA evolution. Characterizing such diagnostic protein profiles, combined with known risk factors as well as emerging genetic data, would contribute to the long-term objective of applying effective non-surgical therapies to the treatment of small AAA disease. Larger studies are required to confirm the potential and clinical role of the identified proteins.
Abstract 226: Thyroid Hormone Stimulated Arteriolar Growth Along With Myocyte Hypertrophy Following Myocardial Infarction In Rats
Authors: Chen Yuefeng , Guo Songchuan , Carrey Zev , Alaie Dariush , Petrillo Richard , Gerdes Anthony M 
Published: July 2021

Background
Thyroid hormones (THs) have been shown to improve LV function and remodeling in animals post-MI and in the human setting, as well as induce capillary and arteriolar growth in the hypertrophic
heart. Whether THs can improve arteriolar growth in the infarcted heart is unknown.

Methods
MI was produced by ligation of the LAD in female SD rats. Rats were divided into the following groups: (1) Sham MI, (2) MI, and (3) MI+T4 treatment (T4 pellet 3.3mg, 60 days release, implanted subcutaneously immediately following MI). Each group was further divided into two subgroups (n=9 per subgroup) for myocytes isolation and whole heart tissues collection eight weeks after surgery. Isolated myocyte shape and arteriolar density in the non-infarcted area were measured at terminal study.

Results
T4 treatment improved LV ±dp/dt, normalized TAU, and increased myocyte cross-sectional area without further increasing myocyte length in MI rats. T4 treatment tended to increase the total length of smaller arterioles (5 to 15 μm) proportional to LV weight increase.

Conclusions
These results suggest that long-term T4 treatment after MI has beneficial effects on myocyte and arteriolar remodeling in the non-infarcted area.
Abstract 270: Acute Reductions in Wall Strain Precede Formation of Intimal Hyperplasia
Authors: Favreau John T , Liu Chengwei , Yu Peng , Mauro Christine , Tao Ming , Gaudette Glenn R , Ozaki C. K 
Published: July 2021

Intimal hyperplasia (IH) remains the major culprit in revascularization failures. We aimed to unravel relationships between acute changes in circumferential arterial wall strain and genesis of IH.

Methods
To induce IH, we employed a validated model using a 9-0 nylon suture tie around the distal mouse common carotid artery (n=10) and an external 35-gauge needle mandrel (OD=0.14mm), with subsequent removal of the mandrel to create a distal common carotid focal stenosis (~78% lumen diameter/~85% flow reduction). Wall strains were measured in three, 1 mm wide regions along the vessel proximal to the focal stenosis at pre-op day 1 and at post-op day 4 (before detectable IH) using Vevo 2100 ultrasonography with VevoVasc software. At post-op day 28, arteries were perfusion fixed and IH was assessed in the same regions as those where strain was analyzed. Strain and morphology were also assessed in the contralateral control artery.

Results
Decreased wall strain was noted in all regions proximal to the focal stenosis from 0.26 ± 0.01 to 0.11 ± 0.02 (p<0.001) with no change in the control artery from pre-op to post-op day 4 (p=0.45). Based on a strain level histogram, vessels were divided into groups with strain ≤0.1 and >0.1. All segments (n = 13) with wall strain ≤0.1 at post-op day 4 had significant IH at day 28. In regions with strains >0.1 at day 4, only 30% had IH at day 28. The average pre-op strains were identical in >0.1 and ≤0.1 strain groups (0.27 ± 0.09 and 0.27 ± 0.08). Mean intimal thickness in vessels with strain ≤0.1 was 32 ± 20 μm, significantly greater than 8.0 ± 16 μm in the group with strain >0.1 (p<0.01). To further understand the mechanisms underlying changes in strain, systolic and diastolic lumen areas were assessed. Although systolic lumen areas in both >0.1 and ≤0.1 groups remained unchanged from pre-op to post-op day 4 (p=0.46), diastolic area was significantly increased in regions with post-op day 4 strain ≤0.1 (p=0.04) but remained unchanged in mice with post-op day 4 strain >0.1 (p=0.67).

Conclusions
Acute reduction in wall strain precedes the formation of IH in this murine model and this change is primarily caused by an increase in diastolic lumen area. Manipulations of wall strain offer a strategy to prevent and attenuate occlusive IH lesions after revascularizations.
Abstract 273: New Reconstruction Technique for MRA to “Fly Through” the Lumen of the Cerebral Vasculature
Authors: Kasam Mallikarjunarao , Ketha Siva S , Konduru Soumya 
Published: July 2021

Methods
Patient exams were performed under an IRB-approved protocol. Two 3D SWIRLS [3] single phase contrast-enhanced angiographies with 1mm3 spatial resolution were used for this work. Clinical exams were acquired on 3.0 T scanners (GE, DVMR 20.1IB, Milwaukee, WI).

Data analysis
The angiogram was post-processed using tree analysis and virtual endoscopy modules of Analyze 11.0 software (Analyze 11.0; Biomedical Imaging Resource, Mayo Clinic, Rochester, MN) to fly through the artery. A statistical parameter called Brightness Area Product (BAP) was defined as the sum of the intensities above the sample minimum intensity/threshold set by the user.

Results and Discussions
Figures (I-IV) show the methodology to fly through the part of anterior carotid artery (ACA) and all statistics of part of the artery (Fig. IV) with angles from the “Root” to its different branches of A1 to G2 (Fig. III). Figure II shows the snap shot of the fly through/internal cross sectional structure of ACA of branch “E2”, at the location marked as “red dot”. Table (IV) represents different statistics including length, angle from the root, area of cross section and the BAP of the branches mentioned in Fig. III. The computation time for this method is < 5 minutes compared to the other reconstruction techniques (>~30 minutes). We proposed a simple and fast automated post-processing method to fly through the artery using Analyze 11.0 software. This technique can be extended to any arteries to monitor internal vasculature and to estimate the stenosis using the statistical parameter BAP, which generates a master profile/database of arteries.

References
1 Shu Y, Bernstein MA, ISMRM 2009 p764.
Abstract 282: Pattern of Development of Abdominal Aortic Aneurysms Induced by Angiotensin In Hypercholesterolemic Mice
Authors: Prins Petra A , Hill Michael , Airey David , Nwosu Sam , Perati Prudhvidhar R , Kon Valentina , Fazio Sergio , Sampson Uchechukwu 
Published: July 2021

Background
Understanding the pattern of development of abdominal aortic aneurysms (AAA) may help optimize translational research strategies. Thus we sought mechanistic insight on the pattern of AAA evolution in apolipoprotein E -/- mice.

Methods
Fifty-nine mice underwent a 4-week pump-mediated infusion of saline (controls; n=13) or 1500 ng/kg/min of AngII (n=46), and AAA development was tracked via in vivo serial ultrasound imaging. We adjusted for hemodynamic covariates in the mixed logistic regression model for AAA occurrence in relation to time.

Results
The incidence of intact AAA in AngII infused mice was 59% (27/46) of which 55.6% (15/27) were small AAA and 44.4% were large AAA (1.2-1.5 vs. >1.5x baseline diameter, respectively). The incidence of rupture was 22% (10/46). All large aneurysms and most ruptures occurred within the first week of AngII exposure. Subsequent aneurysms were small and evolved in the second week of AngII infusion. The overall effect of time was statistically significant (p<0.001) (Figure). When compared to day 7 of the study period, there was no decrease in the log odds of AAA occurrence by day 14 (-0.234, p = 0.65). However, compared to 21 and 28 days into AngII infusion, the log odds of an AAA occurrence decreased by -9.07 (p< 0.001) and -2.35 (p= 0.04), respectively (Figure).

Conclusions
These are the first formal estimates of the odds of AAA occurrence vis-à-vis time in the context of a consistent pattern of AAA development in this mouse model of AngII-induced AAA. They occur in the first (large AAA) and second (small AAA) weeks of AngII infusion, and the odds of occurrence diminish thereafter. These findings provide guidance for experimental strategies.
Abstract 283: Platelet Inhibition Increases Angiotensin II-induced Abdominal Aortic Aneurysm Incidence and Rupture in Mice
Authors: Owens A. Phillip , Boulaftali Yacine , Bergmeier Wolfgang , Luyendyk James P , Mackman Nigel 
Published: July 2021

Objective
Platelets play a central role in both hemostasis and thrombosis. The coagulation protease thrombin activates platelets by cleavage of protease-activated receptors (PAR1 and PAR4 in humans, and PAR3 and PAR4 in mice). Circulating thrombin is increased in patients with abdominal aortic aneurysms (AAAs). We recently demonstrated that PAR4 deficiency in mice increased the incidence of aneurysm (P = 0.001) and rupture-induced death (P = 0.003) in an angiotensin II (AngII) infusion model of AAA. Furthermore, platelet depletion significantly increased rupture in this model (P = 0.048). The purpose of this study was to examine clinically used anti-platelet drugs in this mouse model of AAA.

Methods and Results

Male
Ldlr
-/-
mice (8-12 weeks in age) were fed a fat and cholesterol-enriched diet (21% milk fat, 0.2% cholesterol). Groups of mice received either aspirin (30 mg/L via drinking water [ASA]), or diet supplemented with the direct thrombin inhibitor dabigatran etexilate (10 g/kg chow [DE]) or the P2Y
12
inhibitor clopidogrel (50 mg/kg/day [Plavix]) 1 week prior to and throughout AngII (1,000 ng/kg/min) infusion for 28 days. Drug bioavailability was confirmed with all treatments. Medial diameters in the suprarenal aortic region were increased significantly from baseline to day 28 in all groups infused with AngII, as measured by in vivo ultrasound. Medial diameters were not different in any of the treatment groups compared with placebo controls. However, DE (87% vs. 47%) and Plavix (82% vs. 40%) significantly increased the incidence of AAA versus placebo groups (P < 0.05). ASA also increased the incidence of AAA (93% vs. 70% P = NS). Importantly, all treatments had a significant increase in aortic rupture-induced death versus placebo groups (P < 0.05; DE [67% vs. 7%]; Plavix [41% vs. 0%]; and ASA [64% vs. 10%]). None of the treatments affected total plasma cholesterol, lipoprotein-cholesterol distributions, or AngII-induced increases in systolic blood pressure.

Conclusion
This study indicates that the presence of functional platelets reduces the formation and rupture of AAA in this mouse model. This suggests that inhibition of platelet function may be detrimental to patients with existing AAAs, a conclusion that will be addressed in future mouse studies.
Abstract 294: Expression of Matrix Metalloproteases during the Differentiation of Porcine Adopse-Drived Mesenchymal Stem Cells ro Endothelial Cells
Authors: Almalki Sami G , Rao Velidi , Pankajakshan Divya , Agrawal Devendra K 
Published: July 2021

Rationale

Adipose-derived mesenchymal stem cells (ADMSCs) are multipotent cells that have the potential to differentiate into different cell linages, and represent promising tools in various clinical applications. However, the molecular mechanisms that control the ability of ADMSCs to remodel 3-dimensional extracellular matrix (ECM) barriers during differentiation are not clearly understood. Herein, we studied the expression of matrix metalloproteinases (MMPs) during the differentiation of ADMSCs to endothelial cells (ECs)
in
vitro
.

Methods
MSCs were isolated from porcine abdominal adipose tissue, and characterized by positive staining for MSC markers, CD44, CD73, CD90, and negative staining for CD11b, CD34 and CD45. The plasticity of MSCs was detected by bi-lineage differentiation to osteocytes, and adipocytes. The mRNA transcripts for different MMPs and TIMPs and protein expression of EC markers were analyzed by RT-PCR and immunostaining. The enzyme activity and protein expression were also analyzed by gelatin zymography, ELISA, and Western blot.

Results
The differentiation of ADMSCs to ECs was confirmed by the positive staining and mRNA expression of the endothelial markers. The mRNA transcripts for MMP-2 and membrane type 1 MMP (MT1-MMP) was significantly increased by 2.5 and 2.0 fold, respectively, during the differentiation of MSCs into ECs. Western blot and ELISA showed an elevated MT1-MMP and MMP-2 expression. The enzyme activity of MMP-2 was also observed by gelatin zymography.

Conclusion
We demonstrated that porcine ADMSCs have the ability to differentiate into ECs, and this process involves the up-regulation of MMP-2 and MT1-MMP. The increase in the expression of MMP-2 and MT1-MMP may, at least partially, facilitate the change in morphology of MSCs by degrading the ECM barriers. These findings may provide a potential mechanism for the role of MMP2 and MT1-MMP in the differentiation of ADMSCs into ECs.