bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 494: Rho-a Kinase Activation in Endothelial Cells is Induced by Cocaine or Plasma From Cocaine Consumers: Association With a Prothrombotic Phenotype and Effect of the Inhibitors Atorvastatin and Y-27632
Authors: Saez Claudia G , Pereira-Flores Karla D , Mendoza Camila A , Ocaranza María P , Jalil Jorge E , Massardo Teresa , Mezzano Diego , Pallavicini Julio , Ibañez Carlos , Veliz Jonathan , Ebensberger Roberto , Pereira Jaime 
Published: March 2022

Cocaine consumption is associated with increased risk of cardio- and cerebrovascular diseases, which are the consequence of endothelial dysfunction (ED). We recently showed that cocaine use is related to increased ED markers including circulating endothelial cells, although the mechanisms governing ED in cocaine users are still being researched. Since the RhoA/Rho kinase (ROCK) pathway is involved in the pathogenesis of ED, our
Aim
was to study
in vitro
and
ex vivo
RhoA/Rho kinase activation and ED markers in relation to cocaine consumption.

Method
Plasma samples from active cocaine consumers who met DSM-IV criteria and age and gender-matched controls were tested. ROCK activity, von Willebrand factor (vWf), Metalloproteases (MMP) and platelet adhesion were evaluated in HUVEC supplemented with cocaine (10μM), 10% plasma from cocaine users (CP), normal plasma (NP) or vehicle, in the presence or absence of Y-27632 (10 μM) and atorvastatin (ATS:10μM).

Results

CP induced higher ROCK (p: 0.039) and MMP 2 activity (p<0.05) in HUVEC, released significantly elevated vWf (p0.001), but failed to stimulate MMP activity or vWf release. ATS and Y-27632 significantly reduced ROCK activity (p<0.01 and p<0.001) and platelet adhesion (p< 0.0001 and p: 0.0073) in CP treated cells. ATS also reduced MMP2 activity (p< 0.001) and vWf release (p: 0.004). In cocaine treated cells, ATS and Y-27632 downregulated ROCK activity (p: 0.039 and p: 0.091) and significantly reduced platelet adhesion (p: 0.0156 and p: 0.073).
Ex vivo
studies showed significantly higher ROCK (p 0.015), MMP-2 (p<0.05) and MMP-9 (p<0.01) activity in cocaine consumers than in controls.

Conclusions

Cocaine and cocaine consumer plasma increased ROCK activity in HUVEC, resembling the
ex vivo
findings. Only CP induced higher MMP activity and release of vWf resembling the findings in patients. The significant reduction of most of the in vitro effects by ATS and Y-27632 may provide a novel and potential therapeutic tool for the protection of the endothelium from deleterious stimuli such as cocaine consumption.
Abstract 501: Poly Herbal Formulation Prevents Foam Cell Formation Possibly Through Attenuation of CD36 Scavenger Receptor Signalling Cascade
Authors: Nimgulkar Chetan C , Anand Shankar B , Kumar Uday P , Ghosh Sudip , Kumar Dinesh B 
Published: March 2022

Objective
CD36 signalling cascade promotes migration, spreading and entrapment of lipid laden foam cells within intimal lesions by oxidative stress, inflammation and oxidized LDL (oxLDL) by activation of actin polymerization, Focal Adhesion Kinase (FAK) and cement adhesion which also are indirectly activated by Reactive Oxygen Species (ROS). There is dearth of information about an antagonistic therapy for this CD36 cascading pathway to prevent atherosclerosis. Therefore, we hypothesized that, an antioxidant, antiinflammatory and antihyperlipidemic Poly Herbal Formulation (PHF) consisting of Allium sativum, Zingiber officinale, Curcuma longa, Terminalia arjuna and Cyprus rotundus prevents atherosclerosis.

Methods
Standardized (WHO and IP monographs) hydro-alcoholic extracts of the above said test materials were mixed in equal amounts to prepare PHF. After conforming the antioxidant, antiinflammatory and antihyperlipidemic activities, PHF was evaluated for foam cell formation and the protein expression of CD36, PPAR-γ and FAK in oxLDL treated mouse macrophage J774A.1 cells. Its inhibitory activity for ROS generation was evaluated. Antiatherosclerotic activity was studied in high cholesterol fed rabbit model.

Results
Significant accrual of lipid in oxLDL treated cells conform the foam cell formation where CD36, PPAR-γ and FAK protein levels had increased. PHF (20 to 80 μg/mL) prevented lipid accumulation in oxLDL treated cells from ~29% to ~38% where CD36, PPAR-γ and FAK protein levels were significantly decreased in concentration dependent manner up to 3, 2 and 2 folds, respectively. ROS generation was inhibited by PHF in concentration dependent manner.
PHF dose of 100 and 200mg/Kg, significantly reduced plasma cholesterol, ~23% and ~40% and plasma triglycerides ~87% and ~91%, respectively. Plasma TBARS were significantly reduced by PHF 100 and 200mg/Kg, 50% and ~66%, respectively. Aortic atheroma formation was significantly barred by PHF as observed in histopathological studies.

Conclusion
PHF prevents atherosclerosis by preventing foam cell formation possibly by reducing CD36, PPAR-γ and FAK proteins and ROS generation. However, detailed pharmacodynamic studies are needed to correlate these findings in a clinical set up.
Abstract 502: Arterial Calcification Increases Neointimal Proliferation Following Balloon Injury in Rat Carotid Arteries
Authors: Marshall Andre P , Wang Xue-lin , Paone Darien A , Lin Tonghui , Luo Weifeng , Guzman Raul J 
Published: March 2022

Introduction
Arterial calcification predicts accelerated restenosis after angioplasty and stenting. We sought to determine how calcification affected neointimal accumulation after balloon injury in the rat carotid.

Hypothesis
We propose that transformed SMCs in calcified arteries respond to balloon injury in an accelerated manner through a mechanism involving bone-related factors.

Methods
Arterial calcification was induced in male Sprague-Dawley rats weighing 350-400 grams by subcutaneous administration of vitamin D3. Control rats received vehicle. Both groups underwent left carotid balloon injury one week following injection, a time at which significant medial calcification has developed. Arteries were harvested twenty-one days following balloon injury. The degree of neointimal cell proliferation was next assessed by immunohistochemical staining for PCNA. Neointimal hyperplasia was measured by morphometric analysis using standard imaging analysis software.

Results
At 3 weeks after carotid balloon injury, mean cellular proliferation as measured by PCNA staining was higher in the intima of calcified than control arteries (63.8 ± 6.0 versus 40.5 ± 9.5 cells/hpf, P=ns). Calcified arteries from Vitamin D3-treated rats, however, had significantly more intimal hyperplasia than control uncalcified arteries (intima-to-media ratio = 1.374 ± 0.071 versus 0.758 ± 0.080, P=0.0021). Intimal and medial areas were also individually larger in calcified than control arteries.

Conclusions
Neointimal hyperplasia is accelerated in Vitamin D3-treated rats through a mechanism that partially involves increased SMC proliferation. Calcified vessels may respond differently to balloon angioplasty and this may explain the poorer results of endovascular interventions in patients with arterial calcification. Other factors including accelerated migration and increased matrix synthesis may also be involved. Treatments to inhibit restenosis in calcified arteries may differ from those that work in uncalcified vessels.
Abstract 506: The Effect of Human Recombinant DNase-1 Treatment on Skeletal Muscle Ischemia Reperfusion Injury
Authors: Albadawi Hassan , Oklu Rahmi , Malley Rita , Yoo Hyung-Jin , Austen William G , Watkins Michael T 
Published: March 2022

Objective
Nucleosomes are known to exacerbate tissue injury, thus exogenous DNase-1 has been suggested as a novel therapeutic strategy to limit tissue injury in a stroke model. Our aim was to assess the effect of recombinant DNase-1 treatment on skeletal muscle fiber injury, tissue inflammation and levels of circulating nucleosomes in a mouse model of hind limb ischemia-reperfusion-injury (IR).

Methods
C57BL6 mice were subjected to IR (1.5hr Ischemia/24hr Reperfusion) or sham conditions. Mice were treated with either 50μg DNase-1 IP every 6 hours (DNase-1) or buffer (Control) at the same time intervals. Sham mice were anesthetized for 1.5 hours and allowed to recover for 24 hours. Muscles were harvested to quantify muscle fiber injury, DNAse-1 protein and calcium dependent endonuclease activity. Muscle markers of inflammation Myeloperoxidase (MPO), Keratinocyte chemoattractant (KC) and Plasma nucleosomes were quantified.

Results
Skeletal muscle fiber injury (numbers p<0.05) and endonuclease activity (numbers p<0.001) were greater in the DNase-1 group compared to control and sham. DNase-I localized predominantly to the extracellular matrix and severely injured muscle fibers. There was no significant difference in the plasma levels of nucleosomes or markers of inflammation (MPO or KC).

Conclusions
These novel findings suggest that in contrast to brain IR, hind limb IR increases DNAse-1 activity, and that exogenous treatment with DNase-1 exacerbates muscle injury independent of markers of inflammation. DNase-1 may be a therapeutic target that warrants further investigation.
Table to Be Inserted
Abstract 509: Inhibition of Patched-1 prevents Injury-induced Neointimal Hyperplasia
Authors: Redmond Eileen M , Hamm Katie , Cahill Paul A , Morrow David 
Published: March 2022

Intimal remodeling occurs due to alterations in vascular smooth muscle cell (VSMC) proliferation and apoptosis and is a key event in the pathogenesis of vascular disease. Atherosclerosis, vascular rejection, venous graft restenosis and coronary intervention are each characterized by increased VSMC growth and intimal thickening.

Objectives
To determine the role of Ptc-1 in mediating pulsatile flow-induced changes in SMC growth and neointimal hyperplasia.

Methods and Results
HCASMC were exposed to “normal“ or pathological “low” pulsatile flow conditions for 24 h by utilizing a perfused transcapillary flow system. In vivo, pathological low flow conditions following carotid artery ligation increased medial SMC growth while increasing Ptc-1/Notch expression. Inhibition of Ptc-1 expression by cyclopamine attenuated low flow-induced increases in VEGF/Notch expression while concomitantly decreasing HCASMC growth to normal flow levels. Perivascular delivery of Ptc-1 siRNA by pluronic gel inhibited the low flow-induced increases in Ptc-1/Notch expression and subsequent increase in medial SMC growth. In addition, low pulsatile flow-induced increases in neointimal hyperplasia and subsequent decreased lumen volume were returned to sham-operated control levels following ptc-1 gene knockdown.

Conclusion
These results suggest that pathological low flow stimulates SMC growth and neointimal hyperplasia via Ptc-1 regulation of VEGF/Notch signaling.
Abstract 510: Morphometrics of Calcification in Carotid Endarterectomy Tissue
Authors: Han Richard I , Chen Iou , Brunner Gerd , Morrisett Joel D 
Published: March 2022

Arterial calcification plays a significant role in the pathogenesis of carotid atherosclerotic plaques development. The aim of this study was to provide a detailed analysis of calcification morphology based on histological features in carotid endarterectomy (CEA) specimens. In particular, the emphasis is on calcium nodule size/shape and its relation to tissue and lumen morphologies. These studies will help elucidate the mechanisms of the arterial calcification process.
Representative frozen sections were cut from the common, bifurcation, internal and external segments obtained from 9 CEA samples. Histological stains were used to show the calcified particles, tissue compositions and lipids deposition. The digitized images of sections were warped when necessary to achieve closure of the vessel wall. Image analysis was applied to 2 samples to determine the morphologies of calcium particles (size, shape and perimeter), lumen (area and perimeter) and tissue (area and compositions) using ImageJ. Total non-collagen protein and collagen protein were semi-quantitatively analyzed in one sample.
Calcification occurred predominantly in the bifurcation and internal segments of the CEA, occupying up to 17% of the cross-sectional tissue area. Occassionally, calcification was found in the external branch. The lipids distribution is quite patchy. All calcified particles were contained in the intimal layer of the carotid wall. The lumen area was increased in the bifurcation segment, but it deviated from a patent circular shape. The amount of calcific particles was positively correlated with the tissue area (R2=0.8365, p=0.0015). As the calcium particle increase in size, the shape became more elongated. Collagen accounted for up to 5% of the total non-collagen protein.
Different stages of calcification progression occur at the same time, while the very small sized calcium particle grow in size in all directions, the medium sized particle coalescing with its neighbors to form larger particles. The calcification is accompanied by increase in tissue size and distortion of lumen. Presumably, the kinetics of calcification is determined by the composition of the extracellular matrix and the interstitial cells may play a role in the process.
Abstract 516: Calpain-2 Compensation Promotes Angiotensin II-induced Ascending and Abdominal Aortic Aneurysms in Calpain-1 Deficient Mice
Authors: Subramanian Venkateswaran , Moorleghen Jessica J , Balakrishnan Anju , Howatt Deborah A , Chishti Athar H , Uchida Haruhito A 
Published: March 2022

Background and Objective
We demonstrated that angiotensin II (AngII)-infusion profoundly increased both aortic protein and activity of calpains, calcium-activated cysteine proteases, in mice. In addition, pharmacological inhibition of calpain attenuated AngII-induced abdominal aortic aneurysm (AA) in mice. Recent studies have shown that AngII infusion into mice leads to aneurysmal formation localized to the ascending aorta. However, the precise functional contribution of calpain isoforms (-1 or -2) in AngII-induced abdominal AA formation is not known. Similarly, a functional role of calpain in AngII-induced ascending AA remains to be defined. Using BDA-410, an inhibitor of calpains, and calpain-1 genetic deficient mice, we examined the relative contribution of calpain isoforms in AngII-induced ascending and abdominal AA development.

Methods and Results

Male LDLr-/- mice that were either calpain-1 +/+ or -/- were fed a saturated fat-enriched diet and infused with AngII (1,000 ng/kg/min) for 4 weeks. Calpain-1 deficiency had no effect on body weight or blood pressure during AngII infusion. Intimal areas of ascending aorta and maximum external width of the suprarenal abdominal aorta were measured by en face. Calpain-1 deficiency showed no discernible effects on AngII-induced ascending and abdominal AA development (P = NS). Interestingly, AngII infusion induced increased expression of calpain-2 protein, thus compensating for total calpain activity in aortas of calpain-1 deficient mice. Oral administration of BDA-410 along with AngII infusion significantly attenuated AngII-induced ascending (Vehicle - +/+: 15.0 ± 1.0, -/-: 14.9 ± 0.8; BDA: +/+: 11.5 ± 0.6, -/-: 12.1 ± 0.6 mm
2
; P<0.05) and abdominal (Vehicle - +/+: 1.7 ± 0.1, -/-: 1.6 ± 0.1; BDA: +/+: 1.0 ± 0.1, -/-: 1.1 ± 0.1 mm; P<0.05) AA formation in both calpain-1 +/+ and -/- mice as compared to vehicle administered mice. Western blot and immunostaining analyses revealed BDA-410 administration attenuated AngII-induced C-terminal fragmentation of filamin A (P<0.05), an actin binding cytoskeletal protein, and medial macrophage accumulation in aorta.

Conclusion
Calpain-2 compensates for loss of calpain-1, and promotes AngII-induced ascending and abdominal AAs in calpain-1 deficient mice.
Abstract 534: Superoxide Dismutase 1 Modulates STAT3 pathway in Angiotensin II Induced Vascular Remodeling
Authors: Shiraishi Yasunaga , Kujiraoka Takehiko , Niida Tomiharu , Ishigami Norio , Arai-Nakaya Yuko , Hakuno Daihiko , Isoda Kikuo , Adachi Takeshi 
Published: March 2022

[Background]
Superoxide dismutase 1 (copper/zinc SOD; SOD-1) generates hydrogen peroxide (H2O2) from superoxide and is predominantly expressed in cytosol of vascular walls. Angiotensin II (Ang-II) induces vascular remodeling is associated with H2O2 generation from NADPH oxidase. We have previously shown that SOD-1 modulates Ang-II induced vascular remodeling. The aim of this study is to seek the signaling mechanisms for SOD-1to modulate vascular remodeling with Ang-II in in vivo.

[Methods and Results]
We employed SOD-1 deficient mice (SOD-1-/-), which were continuously infused with Ang-II (3.2mg/kg/day) for seven days. There was no difference in blood pressure between SOD-1-/- and their wild-type mice (WT). The serum levels of derivatives of reactive oxygen metabolites were similarly increased in both WT and SOD-1-/- with Ang-II, however, an increase in oxidized glutathione levels in whole blood cells were blunted in SOD-1-/-, suggesting a decrease in cytosolic H2O2 generations with SOD-1 deficiency. In histological analysis, Ang-II increased aortic wall thicknesses in WT, however, the responses were also blunted in SOD-1-/- (wild-type: 91.9±3.0μm SOD-1-/-: 68.4±4.5μm, P<0.001, N=10-12). In western blot analysis, Ang-II infusion enhanced the phosphorylation of signal-transducer-and-activator-of-transcription 3 (STAT3) in aortas from WT, which was blunted in SOD-1-/-, while the phosphorylation of ERK and Akt were identical. The interleukin 6 (IL-6) of serums were equally elevated in both SOD-1-/- and WT with Ang-II infusion. In cultured rat vascular smooth muscle cells (VSMCs), Ang II (24 h) slowly increased, and IL-6 (5 min) or hydrogen peroxide (H2O2) (15min) rapidly increased the phosphorylation of STAT3. With copper-chelating agents (N, N-diethyldithiocarbamate; 1mM 4h), the phosphorylation of STAT3 was blunted with all agonists.

[Conclusion]
These results indicated that the activation of STAT3 plays a role in Ang II- induced vascular remodeling, and SOD-1 is involved with the signaling mechanism in in vitro and in vivo.
Abstract 537: Platelet Derived Growth Factor Induced Vascular Smooth Muscle Cell Proliferation is Associated with Enhanced Glut1 Expression/glycosylation and Glucose Uptake
Authors: Poulose Ninu , Pyla Rajkumar , Osman Islam , Ganapathy Vadivel , Segar Lakshman 
Published: March 2022

Objective

The role of glucose transporters (GLUTs) in the phenotypic regulation of vascular smooth muscle cells (VSMCs) has not been fully characterized. The present study has examined the effects of PDGF (
vs
insulin) on GLUT isoform expression/localization, GLUT1 glycosylation, glucose uptake and ATP formation during VSMC proliferation.

Methods
Human aortic VSMCs were exposed to PDGF (30 ng/ml) or insulin (1 μM) for different time intervals. GLUT isoform expression was determined by qPCR and immunoblot analysis. GLUT isoform distribution/localization was determined by subcellular fractionation studies and immunocytochemical analysis. Glucose utilization and ATP formation were determined by glucose oxidation assay and luminescent ATP detection assay, respectively. The role of GLUT1 in VSMCs was determined by GLUT1 siRNA knockdown studies.

Results

Exposure of VSMCs to PDGF resulted in significant increases in glucose uptake and cell proliferation, compared with insulin. In addition, PDGF and insulin enhanced the mRNA expression levels of total GLUT isoforms (GLUT1 through 14) to ~4317/10
5
B2M
transcripts and ~8,174 transcripts/10
5
B2M
transcripts, respectively, compared with control (~3259 transcripts/10
5
B2M
transcripts). In particular, PDGF and insulin enhanced GLUT1 mRNA to 1711 ± 38/10
5
B2M
transcripts and 3387 ± 58 transcripts/10
5
B2M
transcripts, respectively. Immunoblot analysis using whole cell homogenates showed a significant increase in GLUT1 protein expression in PDGF or insulin-treated VSMCs. Importantly, subcellular localization studies using PDGF-treated VSMCs revealed enhanced GLUT1 protein expression and its glycosylation in the plasma membrane. Furthermore, PDGF enhanced glucose metabolism as revealed by increased glucose utilization and ATP formation. Down regulation of GLUT1 using target-specific siRNA significantly attenuated PDGF-induced glucose uptake.

Conclusions
Enhanced expression and glycosylation of plasma membrane-localized GLUT1 isoform plays a major role in promoting VSMC proliferation, which may contribute to accelerated atherosclerosis.
Abstract 545: Response Gene to Complement 32-deficiency Causes Impaired Placental Angiogenesis in Mice
Authors: Cui Xiaobing , Guo Xia , Chen Shiyou 
Published: March 2022

Aims
The objectives of this study are to determine the role of response gene to complement 32 (RGC-32) in the placental angiogenesis during pregnancy and explore the underlying mechanisms.

Methods and Results
RGC-32-deficient (RGC32-/-) mice were generated from C57BL/6 embryonic stem cells with deletion of Exon 2 and 3 of RGC-32 gene. Most of the RGC32-/- mice can survive. However, their body sizes were much smaller compared to their wild-type littermates when they were born. By examining the embryo development and placentas at 16.5 days post-coitum, we found that RGC32-/- embryos and fetal placentas were significantly smaller than the wild-type. Further analysis showed that the labyrinth zone of RGC32-/- fetal placenta was smaller with defective angiogenesis. Mechanistically, vascular endothelial growth factor (VEGF) receptor 2 (VEGFR2) and placental growth factor (PlGF), but not other angiogenic factors, were significantly downregulated in RGC32-/- placentas, suggesting that VEGFR2 and PlGF may mediate RGC-32 function in placenta angiogenesis. Indeed, knockdown of RGC-32 by shRNA inhibited VEGF-induced endothelial cell proliferation, migration, and tube formation while blocking VEGFR2 expression. Knockdown of RGC-32 also inhibited trophoblast proliferation while blocking the expression of PlGF.

Conclusion
Absence of RGC-32 caused intrauterine growth restriction through interrupting the placental angiogenesis, which was due to the decreased expressions of VEGFR2 in endothelial cells and PlGF in trophoblast cells.