bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 218: Orphan Nuclear Receptor Nur77 is a Novel Negative Regulator of ET-1 Expression in Vascular Endothelial Cells
Authors: Qin Qing , Chen Ming , Yi Bing , Li Pan , You Xiaohua , Sun Jianxin 
Published: March 2022

Objective
Endothelin-1 (ET-1), a vasoactive peptide mainly produced by vascular endothelial cells, plays an important role in the regulation of vascular tone and the development of cardiovascular diseases. The objective of the present study is to identify the novel regulators implicated in the regulation of ET-1 expression in vascular endothelial cells (ECs), and to determine the molecular mechanism involved.

Method and Results
by using quantitative real-time PCR (qRT-PCR) and Enzyme-linked immunosorbent assay (ELISA), we showed that either ectopic expression of Nur77 or pharmacological activation of Nur77 by 6-mercaptopurine (6-MP) substantially inhibited the ET-1 expression, under both basal and thrombin stimulated conditions, in human umbilical vein endothelial cells (HUVECs). Furthermore, knockdown of Nur77 enhances both basal and thrombin induced ET-1 expression, suggesting that Nur77 is a negative regulator of the ET-1 expression in vascular ECs. Inhibition of ET-1 expression by Nur77 mainly occurs at the gene transcriptional levels, since Nur77 potently inhibited the ET-1 promoter activity, without affecting the ET-1 mRNA stability. As shown in electrophoretic mobility shift assay (EMSA), Nur77 over-expression substantially inhibited both basal and thrombin stimulated transcriptional activity of AP-1, which is a critical transcriptional factor implicated in the ET-1 expression in ECs. Mechanistically, we found that Nur77 specially interacts with c-Jun in the nuclear fraction of endothelial cells by immunoprecipitation and inhibits the AP-1 dependent c-Jun promoter activity, which eventually leads to the decreased expression of c-Jun, a critical component for both AP-1 transcriptional activity and ET-1 expression in ECs.

Conclusions
These results demonstrate for the first time that Nur77 is a novel negative regulator of the ET-1 expression in vascular ECs through an inhibitory interaction with c-Jun/AP-1 pathway. Targeting Nur77 may represent a potentially novel therapeutic strategy for preventing certain cardiovascular diseases, such as atherosclerosis, vascular complications of diabetes, and pulmonary artery hypertension.
Abstract 228: Distinct Roles of PDGF-C and VEGF in Impaired Angiogenesis of Diabetes and Its Implication in Therapeutic Angiogenesis
Authors: Moriya Junji , Ferrara Napoleone 
Published: March 2022

Background

Platelet-derived growth factor-C (PDGF-C) is known to promote angiogenesis independently of vascular endothelial growth factor (VEGF), although its significance in postnatal angiogenesis
in vivo
remains poorly understood.

Methods and Results
We employed murine model of hind limb ischemia and found that expression of PDGF-C and its receptor PDGFR-alpha were markedly up-regulated in ischemic limbs. Treatment with a neutralizing antibody against PDGF-C significantly impaired blood flow recovery and neovascularization after ischemia almost to the same extent as treatment with a VEGF neutralizing antibody. Moreover, PDGF-C- deficient mice showed a reduced blood flow recovery after ischemia compared to wild-type mice, confirming a strong pro-angiogenic activity of PDGF-C.
Next we injected an expression vector encoding the PDGF-C gene into ischemic limbs. Blood flow recovery and neovascularization after ischemia were significantly improved in the PDGF-C-treated group compared with the control group.
It has been reported that angiogenic response to ischemia is attenuated in diabetes even after VEGF treatment, although a precise mechanism remains unknown. We hypothesized that PDGF-C might relate to the impaired angiogenesis of diabetes. To test this hypothesis, we created murine model of diabetes by intraperitoneal injection of streptozotocin (STZ). We found that expression levels of PDGF-C at baseline and after ischemia were both significantly lower in limb tissues of diabetic mice than in those of control mice, while expression levels of VEGF and other members of the PDGF family were not changed or even higher in diabetic mice. Unlike in non-diabetic mice, introduction of VEGF gene into ischemic limbs did not improve blood flow recovery in diabetic mice. On the other hand, these changes were effectively reversed by introduction of the PDGF-C gene.

Conclusion
These results indicate that down-regulation of PDGF-C expression in limb tissues of diabetic mice contribute to impaired angiogenesis and suggest that introduction of PDGF-C would be a novel strategy for therapeutic angiogenesis, especially when VEGF treatment is ineffective, such as in the diabetic state.
Abstract 238: Aortic Intima-Media Thickness by M-mode or 2-Dimensional Semi-Automated Imaging: Do These Methods Provide Equivalent Results?
Authors: Macias Leonardo , Schevchuck Alex , Roldan Paola C , Qualls Clifford R , Sibbitt Wilmer L , Middendorf Julia , Rodan Carlos A 
Published: March 2022

Background
Aortic (Ao) atherosclerosis is best defined by manual quantitation of intima-media thickness (IMT) using M-mode. However, M-mode imaging is time consuming and requires experience. Thus, we aimed to determine if Ao IMT assessed by M-mode and 2-dimensional (2-D) semi-automated imaging provide similar results.

Methods
57 volunteer subjects (51 women, age 36±12 years) underwent multiplane transesophageal echocardiography (TEE) using a 7 MHz phased array transducer with a 0.1 mm axial resolution. At low depth (3-4cm) and with a narrow sector scan to improve image resolution, 2-D guided M-mode images were obtained to assess IMT of the anterior wall of the proximal, mid, and distal descending thoracic Ao. At each Ao level, 3 IMT measurements were performed from Ao long axis views during end-diastole. Studies were measured off-line using electronic calipers by an experienced observer. A second observer unaware of M-mode results and using 2-D semi-automated imaging performed Ao IMT measurements from long axis images at each Ao level and same site where M-mode measurements were obtained.

Results

IMT of proximal, mid, and distal Ao by M-mode was similar to that obtained by 2-D semi-automated imaging (p≥0.19) (
Table
). Less variability of IMT values occurred by 2-D than by M-mode imaging (coefficient of variation 17% vs 38%). Both imaging methods were highly correlated (overall r=0.69, p<0.001).

Conclusion
Assessment of Ao IMT by TEE M-mode and 2-D semi-automated imaging provide equivalent results.
Abstract 250: Surface Microscale Topograhical Features Promotes Endothelial Cell Proliferation and Migration on a Nanocomposite Vascular Graft Material
Authors: Chong Debra S , Dalby Matthew J , Gadegaard Nikolaj , Seifalian Alexander M , Hamilton George 
Published: March 2022

Objectives
Micro and nanoscale surface topographical features have been shown to have an influence on cellular behaviour. Endothelisation of small diameter vascular grafts has long been thought to have beneficial and protective effects as well as ensuring patency of the graft. Our objective is to explore the potential of using surface topographical features on a biocompatible nanocomposite polymeric material to produce a ‘self-endothelising’ vascular graft.

Methods
Photolithography was used to fabricate microscale topographical grooves onto a mask. These features were then heat-casted onto a nanocomposite polymeric material made of polyhedral oligomeric silsesquioxane (POSS) and poly (carbonate urea) urethane (PCU). Fidelity of the microscale grooves were confirmed using scanning electron microscopy (SEM) and atomic force microscopy (AFM). Human umbilical vein endothelial cells (HUVECs) were seeded onto POSS-PCU sheets with different microscale grooves and cellular behaviour was monitored daily. Cell proliferation and metabolic activity was measured using a colorimetric assay. Adhesion of the cells were confirmed using fluorescence microscopy.

Results
Our results show that grooves can be reproduced with high fidelity using masks produced by photolithography onto POSS-PCU nanocomposite polymer. Ridges of width 12.5 μm with channels widths of 25μm and subsequent depths of 300 - 700 nm were measured and confirmed using SEM and AFM These ordered grooves enhance cell spreading and adhesion compared with flat polymer surfaces. Furthermore, cell proliferation is further improved on these microscale features compared to featureless surfaces.

Conclusion
Microscale grooves on the surface of a nanocomposite polymeric material promotes increased endothelial cell adhesion, migration and proliferation. These ordered topographical features could be used to produce a new generation of vascular grafts with ‘self-endothelising’ potential.
Abstract 264: Edaravone Injected at the Start of Reperfusion Suppresses Lung Damage Caused by Myonephropathic Metabolic Syndrome in Rats
Authors: Yamamura Mitsuhiro , Mitsuno Masataka , Tanaka Hiroe , Ryomoto Masaaki , Fukui Shinya , Miyamoto Yuji 
Published: March 2022

Introduction & Hypothesis
Free radicals have been implicated in myonephropathic metabolic syndrome (MNMS), which damages not only muscles but also lungs. At ATVB 2012, we previously reported that radical scavenger, edaravone (Radicut®, Mitsubishi Tanabe Pharma Co., Japan) injected at the start of reperfusion suppresses muscle injury following leg ischemia in rats. In this study, we evaluated whether edaravone can suppress MNMS-induced lung damage following leg ischemia, if injected at the start of reperfusion.

Methods
The MNMS models (Male Lewis rats, 525 ± 78 g, n = 10) were induced, by clamping the bilateral common femoral arteries for 5 hours and then de-clamping. The rats were intraperitoneally injected with either 9.0 mg / kg of edaravone (edaravone group; n = 5), or saline (MNMS group; n = 5) at the same time as de-clamping, corresponding to the start of reperfusion. Five hours after de-clamping, the both lungs were harvested and stained with hematoxylin & eosin (HE). Normal lungs without reperfusion were also harvested as control (n = 3). Lung damage was expressed as the percentage area of the alveolar wall thickness, determined by computerized densitometry (NIH Image Program J, Ver. 1.37).

Results
The lungs of MNMS group had extensive swelling. But the lungs of the edaravone group were filled with air, similar to that of normal lungs (Figure attached, original 200x). The mean percentage area in the edaravone group was significantly lower than that in MNMS group (17.2 ± 5.2 vs. 63.6 ± 5.6%, p < 0.001).

Conclusions
Our results suggest that edaravone injected at the start of reperfusion can suppress MNMS-induced lung damage following leg ischemia in rats.
Abstract 271: Scavenger Receptor Bi is a Key Determinant and HDL is a Critical Modulator of Thymocyte Apoptosis in Sepsis
Authors: Li Xiang-an , Guo Ling , Zheng Zhong , Ai Junting , Howatt Deborah , Daugherty Alan 
Published: March 2022

Background-
Thymocyte apoptosis is a major event in sepsis; however, how this process is regulated remains poorly understood.

Methods and Results-

We induced sepsis in scavenger receptor BI (SR-BI) null and wild type littermates with cecal ligation and puncture (CLP). CLP induced profound thymocyte apoptosis in SR-BI
+/+
mice, but no thymocyte apoptosis in SR-BI
-/-
mice, indicating that SR-BI is a key determinant of thymocyte apoptosis in sepsis. SR-BI controlled inducible glucocorticoid (GC) generation during sepsis, which is required for triggering thymocyte apoptosis. Unexpectedly, supplementation with GC only partly restored thymocyte apoptosis in SR-BI
-/-
mice. We demonstrated that HDL is a critical modulator for thymocyte apoptosis. SR-BI
+/+
HDL significantly enhanced GC-induced thymocyte apoptosis but SR-BI
-/-
HDL had no such activity. Further study revealed that SR-BI
+/+
HDL modulates GC-induced thymocyte apoptosis via promoting glucocorticoid receptor translocation, but SR-BI
-/-
HDL loses such regulatory activity. To understand why SR-BI
-/-
HDL loses its regulatory activity, we analyzed HDL cholesterol contents and found that there was 3-fold enrichment of unesterified cholesterol in SR-BI
-/-
HDL compared with SR-BI
+/+
HDL. Importantly, normalization of unesterified cholesterol in SR-BI
-/-
HDL by probucol administration restored GC-induced thymocyte apoptosis both
in vitro
and
in vivo
, and incorporating unesterified cholesterol into SR-BI
+/+
HDL rendered SR-BI
+/+
HDL dysfunctional.

Conclusions-
The findings in this study reveal a previously unrecognized regulatory mechanism of thymocyte apoptosis in sepsis, involving SR-BI as a key determinant and HDL as a critical modulator.
Abstract 274: HDL-Mediated Cellular Cholesterol Efflux in Patients with Severe Carotid Atherosclerosis
Authors: Hafiane Anouar , Doonan Robert J , Daskalopoulou Styliani , Genest Jacques 
Published: March 2022

The objective of this study was to examine the function of high-density lipoproteins (HDL) in patients with severe carotid atherosclerosis undergoing carotid endarterectomy (CEA) and in patients with coronary artery disease (CAD).
We examined clinical, biochemical and lipoprotein profiles and HDL cholesterol efflux capacity (CEC) in 155 patients undergoing CEA and compared the results with patients with an acute coronary syndrome (ACS time 0 n=26) measured within 48 hours of the event, and again at 12 weeks (ACS recovery, n=26), in patients with chronic, stable CAD (sCAD) n=27 and in healthy controls. There were 110 men (71%) and 45 women (29%) with a mean age 69±10 years in the CEA group. Mean HDL-C was 0.96±0.26 mmol/L (38±10 mg/dL). There were no significant differences in HDL-C between CEA, ACS and sCAD subjects.
HDL was obtained after depletion of apo B-containing lipoproteins with PEG precipitation. Cellular efflux capacity was determined by incubating HDL in cAMP-stimulated J774 mouse peritoneal macrophages for 6 hours. Specific cholesterol efflux was obtained by subtracting total efflux from efflux in non-cAMP stimulated cells. The procedure was standardized to enable medium throughput. Coefficient of variability was approximately 5%.
HDL from CEA patients differ in its ability to promote cholesterol efflux. The range of CEC observed was: min 14.7%, max 34.0% mean 22.4±0.6%; (IQR 18.9-27.3% median 22.0). We found a weak correlation between HDL-C and CEC in patients CEA (r= 0.20, P=0.012); a significant correlation was found between apo A-I and CEC (r=0.23, P=0.005). Subjects with similar HDL-C or apoA-I differed in their ability to promote cellular cholesterol efflux, suggesting that the HDL-C mass does not reflect functionality. We found similar CEC in patients with CAD and ACS recovery.
Here, we report that patients with severe carotid atherosclerosis have low HDL-C and a CEC similar to that observed in CAD patients. The function of HDL to promote cellular cholesterol efflux, as determined by CEC, varied over a wide range and was only weakly correlated with HDL-C. The significant correlation between apo AI and CEC suggests that HDL particle size -and composition may also be an important determinant of the ability to promote cellular cholesterol efflux.
Abstract 281: Cyclophilin A is a Novel Biomarker for Oxidative Stress and Atherosclerotic Diseases
Authors: Satoh Kimio , Fukumoto Yoshihiro , Sugimura Koichiro , Miura Yutaka , Aoki Tatsuo , Nochioka Kotaro , Tatebe Shunsuke , Tatebe Shunsuke , Yamamoto Saori , Shimizu Toru , Takagi Yusuke , Tsuburaya Ryuji , Ito Yoshitaka , Matsumoto Yasuharu , Nakayama Masaharu , Takeda Morihiko , Takahashi Jun , Ito Kenta , Yasuda Satoshi , Shimokawa Hiroaki 
Published: March 2022

Background
We have recently demonstrated that oxidative stress induces secretion of cyclophilin A (CyPA) from vascular smooth muscle cells, which plays a crucial role in the pathogenesis of atherosclerosis and abdominal aortic aneurysms (AAA) in mice. Thus, we tested our hypothesis that plasma CyPA levels could be a biomarker of atherosclerotic diseases such as coronary artery disease (CAD) and AAA in humans.

Methods and Results
In consecutive 320 patients undergoing coronary angiography, we examined the relationship between the severity of CAD and plasma CyPA levels measured by an immunoassay. Plasma CyPA levels were significantly higher in patients with significant coronary stenosis (>50%, n=189) compared to those without it (n=131) (P<0.001). A significant positive correlation was noted between plasma CyPA levels and significant coronary stenosis even after adjustment for age, sex, hypertension, diabetes, dyslipidemia and smoking. The average number of coronary arteries with significant stenosis and the need for coronary intervention were significantly increased in the quartiles of higher CyPA levels (both P<0.001). In addition, plasma CyPA levels were significantly correlated with the number of coronary arteries with significant stenosis (P<0.001). Indeed, plasma CyPA levels were a strong predictor of CAD (adjusted odds ratio for CAD, 6.20; 95% confidence interval, 3.14-12.27; P<0.001). Interestingly, plasma levels of CyPA increased according to the number of atherosclerotic risk factors that enhance oxidative stress. Furthermore, plasma CyPA levels significantly reduced after medical treatment of those risk factors. Finally, pathological examinations showed that CyPA was strongly expressed in atherosclerotic coronary plaque of patients with myocardial infarction. In a separate study, we examined the plasma CyPA levels in patients with aortic aneurysms. Plasma CyPA levels were significantly higher in patients with aortic aneurysms (n=58) compared to those without it (n=69) (P<0.001). Importantly, plasma CyPA levels were significantly higher in patients coexisting with CAD (n=18) compared to those only with aortic aneurysms (n=40) (P=0.012).

Conclusion
Plasma CyPA level is a novel biomarker of atherosclerotic diseases.
Abstract 288: Differences in Proliferative and Migratory Capability of Human Myoblasts Isolated From CLI and Control Patients
Authors: Yu Rebekah Sian Hwee , Abraham David , Baker Daryll , Tsui Janice 
Published: March 2022

Objectives
Patients with critical limb ischemia (CLI) have attributable myopathy that impairs their functional ability as well as dampening the positive effect of successful revascularisation outcomes. Adult skeletal muscle retains the potential to repair following different injury patterns. We aim to investigate the potential differences in proliferation and migration capability of human myoblasts with CLI and asymptomatic control patients.

Methodology
Gastrocnemius muscle biopsies were obtained from patients with critical limb ischaemia and control samples were obtained from non-ischaemic patients. Human myoblasts were isolated and cultured from each sample and stained with the myoblast marker desmin. Confirmed myoblast cultures (<5% alternative cell type contamination) were then used to investigate the proliferative capability (MTT assay) as well as migratory potential (scratch-wound assay) under both ischaemic and normoxic conditions of each group.

Results
Myoblasts isolated from CLI patients demonstrated greater proliferative ability in comparison to control samples. However, control samples revealed greater capacity to heal scratch wounds made in a monolayer of myoblasts in both normoxia and hypoxic conditions.

Conclusion
Isolation of human myoblasts, especially from disease groups provides a useful platform to perform in vitro analysis to better understand and characterise pathology. CLI myoblasts exhibited enhanced proliferative but diminished migratory potential in comparison to control myoblasts.
Abstract 30: Leveraging the Innate Immunity Pathway for Transdifferentiation of Fibroblasts to Endothelial Cells
Authors: Sayed Nazish , Wong Wing Tak , Cooke John P 
Published: March 2022

Background
Cell-based approaches to regenerate the endothelium holds promise, with one such candidate source being induced pluripotent stem cells (iPSCs). We have investigated the potential of endothelial cells (ECs) derived from iPSCs to promote the perfusion of ischemic tissue in a murine model of peripheral artery disease (PAD). However, it may be more efficient to transdifferentiate fibroblasts to ECs directly. Recent reports have suggested that direct reprogramming to ECs is feasible, however still requires the use of viral vectors encoding transcription factors, thus making them clinically unsafe.

Hypothesis
We recently discovered that retroviral vectors encoding the reprogramming factors, by activating the Toll-like receptor 3 (TLR3) pathway, make nuclear reprogramming possible by increasing epigenetic plasticity and favoring an open chromatin state (Lee & Sayed et al. Cell). Based on this recognition that innate immunity favors an open chromatin state, we hypothesized that activation of TLR3, together with external microenvironmental cues that drive EC specification, might induce transdifferentiation of fibroblasts into ECs (“iECs”).

Results
Intriguingly, our preliminary data showed that TLR3 agonist Poly I:C, combined with exogenous endothelial growth factors, was sufficient to transdifferentiate fibroblasts into iECs (in the absence of viral vectors or transcription factors). These iECs exhibited all the characteristics of EC phenotype comparable to HMVECs including ability to form capillary-like structures and incorporating acetylated-LDL. Moreover, loss-of function studies showed that activation of TLR3 plays a role in the efficient transdifferentiation of human cells when using Poly I:C. Furthermore, iECs significantly improved limb perfusion and neovascularization in the ischemic hindlimb compared to control groups.

Conclusion
This study is a first step toward development of a regenerative strategy for PAD on the use of ECs derived from small molecules and growth factors without use of viral vectors encoding transcription factors. Moreover, it has allowed us to generate a small molecule strategy for therapeutic transdifferentiation that might be applied for direct reprogramming in vivo.