bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 22: OSBPL6 is a Novel miR-33 Target Gene Involved in Cellular Cholesterol Efflux
Authors: Hennessy Elizabeth J , Rayner Katey J , Sokolovska Anna , Ouimet Mireille , Esau Christine , Moore Kathryn J 
Published: July 2021

Cellular lipid homeostasis is fundamental to human health. Recent studies indicate that microRNAs (miRNAs) play important roles in the post-transcriptional regulation of lipid metabolism genes. This is illustrated by the discovery, by our group and others, of miR-33 as a central regulator of multiple aspects of lipid metabolism, including cholesterol efflux, fatty acid oxidation, and plasma levels of HDL cholesterol and VLDL triglycerides. In a study of miR-33 antagonism in non-human primates, we identified the most highly de-repressed miR-33 target gene as OSBPL6 (oxysterol binding protein like 6). A member of the OSBP family of cytosolic proteins that bind cholesterol/oxysterols, OSBPL6 contains an ER-targeting FFAT motif as well as a predicted plasma membrane-targeting pleckstrin homology domain, suggesting a putative role for OSBPL6 in vesicular traffic between these compartments. Notably, the OSBPL6 gene resides in a chromosome 2 locus associated with a predisposition to CAD in two major human heart disease studies (Family Heart Study, Framingham Heart Study). We therefore sought to further understand the regulation and function of OSBPL6 in cellular lipid homeostasis.

Results
We show that OSBPL6 is induced in macrophages and hepatocytes in response to cholesterol loading in vitro, as well as in the livers of LDLR-/- mice and non-human primates fed a high fat diet. Overexpression of miR-33 reduces OSBPL6-3’UTR activity by 40%, and this repression is relieved by mutation of the putative miR-33 binding site in the 3’UTR, confirming that OSBPL6 is a direct target of miR-33. Accordingly, transfection of HEPG2 cells with miR-33 mimic decreases mRNA expression of Osbpl6, but not the related family member Osbpl1. Notably, siRNA-mediated knockdown of OSBPL6 in cholesterol-loaded HEPG2 cells or THP-1 macrophages impairs cholesterol efflux to both apoA-I and HDL, and conversely, OSBPL6 overexpression enhances cholesterol efflux to these acceptors.

Conclusion
These data identify OSBPL6 as a novel miR-33 target gene in mice and humans that regulates cellular cholesterol trafficking and efflux via the ABC transporters, further highlighting the coordinated regulation by miR-33 of pathways that promote cellular cholesterol clearance.
Abstract 222: Dkk1 and Msx2-Wnt7b Signaling Reciprocally Regulate the Endothelial-Mesenchymal Transition in Aortic Endothelial Cells
Authors: Cheng Su-Li , Shao Jian-su , Behrmann Abraham , Krchma Karen , Towler Dwight A 
Published: July 2021

Objective
Endothelial cells (ECs) can undergo an endothelial-mesenchymal transition (EndMT) during tissue fibrosis. Wnt- and Msx2-regulated signals participate in arteriosclerotic calcification and fibrosis. We studied the impact of Wnt7, Msx2, and Dkk1 (Wnt7 antagonist) on EndMT in primary aortic endothelial cells (AoECs).

Methods and Results
Transduction of AoECs with vectors expressing Dkk1 suppressed EC differentiation and induced a mineralizing myofibroblast phenotype. Dkk1 suppressed claudin 5, PECAM, cadherin 5 (Cdh5), Tie1 and Tie2. Dkk1 converted the cuboidal cell monolayer into a spindle-shaped multilayer and inhibited EC cord formation. Myofibrogenic and osteogenic markers - e.g., SM22, type I collagen, Osx, Runx2, alkaline phosphatase – were upregulated by Dkk1 via activin-like kinase / Smad pathways. Dkk1 increased fibrosis and mineralization of AoECs cultured under osteogenic conditions - the opposite of mesenchymal cell responses. Msx2 and Wnt7b maintained the “cobblestone” morphology of differentiated ECs and promoted EC marker expression. Deleting EC Wnt7b with the Cdh5-Cre transgene in Wnt7b(fl/fl);LDLR-/- mice upregulated aortic osteogenic genes (Osx, Sox9, Runx2, Msx2) and nuclear pSmad1/5, and increased collagen accumulation.

Conclusions
Dkk1 enhances EndMT in AoECs, while Msx2-Wnt7 signals stabilize EC phenotype. EC responses to Dkk1, Wnt7b, and Msx2 are the opposite of mesenchymal cell responses, coupling EC phenotypic stability with osteofibrogenic predilection during arteriosclerosis.
Abstract 247: Association of Soluble RAGE Levels with Carotid Atherosclerosis: The Northern Manhattan Study (NOMAS)
Authors: Hudson Barry I , Gardener Hannah , Liu-Mares Wen , Dong Chuanhui , Cespedes Sandino , Elkind Mitchell S , Wright Clinton B , Sacco Ralph L , Rundek Tatjana 
Published: July 2021

Introduction
The Receptor for Advanced Glycation End-products (RAGE) is a multi-ligand receptor that propagates vascular cell dysfunction leading to proinflammatory disease states. RAGE is produced as a membrane bound and soluble isoform (soluble RAGE (sRAGE)), with the soluble isoform demonstrated to act as a RAGE decoy preventing cellular activation and atherosclerosis. Recent cohort studies have suggested that serum levels of sRAGE are associated with the risk of CVD and therefore may be a novel biomarker for cardiovascular disease states.

Hypothesis
We hypothesized that sRAGE levels are associated with subclinical atherosclerosis in an ethnically diverse population.

Methods
We included 1,102 stroke-free participants from the multi-ethnic Northern Manhattan Study (NOMAS) who underwent high-resolution carotid B-mode ultrasound to measure carotid plaque phenotypes (density, thickness, and area) and carotid intima-media thickness (IMT). Plaque density was characterized by Gray Scale Median (GSM). Serum sRAGE was measured by ELISA and log-transformed to stabilize variance. Multiple linear and logistic regressions were employed to estimate sRAGE associations with IMT and plaque measures.

Results
The mean age at time of ultrasound was 70.7±8.6yrs; 65% were Hispanic, 19% black, and 16% white. The majority of subjects had carotid plaque present (54%) with the median GSM 38(0-190). Mean plaque thickness (IQR) was 1.30(0-3.99)mm and mean area (IQR) 2.43(0-96.75)mm2. Mean IMT was 0.93±0.09mm. High sRAGE levels were associated with more echolucent plaques (OR 1.2, 95% CI 1.03-1.42), especially among Hispanics (OR 1.26, 95% CI 1.04-1.54). These relationships remained after adjusting for sociodemographic and vascular risk factors. No association was seen between sRAGE levels and carotid IMT, plaque thickness or area.

Conclusion
In the present study, higher sRAGE levels were associated with echolucent (lower density) plaque, especially among Hispanic subjects. Our data suggest sRAGE levels may be associated with atherosclerotic plaque morphology and its vulnerability, especially among minority groups. These findings further support RAGE as a novel target for anti-atherosclerosis interventions.
Abstract 268: Revascularization with Open Bypass vs. Angioplasty and Stenting of the Lower extremity Trial (ROBUST)
Authors: Malas Mahmoud B , Qazi Umair , Black James , Reifsnyder Thomas , Freischlag Julie , Perler Bruce 
Published: July 2021

Background
Peripheral Arterial Occlusive Disease (PAD) is the most common vascular condition affecting 3-10% of the general population, and as high as 15-20% of those over 70 years of age. PAD is also a major marker for cardiovascular disease and stroke.
The Transatlantic Inter-Society Consensus (TASC II) classified superficial femoral artery lesions into 4 categories based on length and extent of disease. For TASC II A (short) lesions angioplasty with stenting (PTA/S) has shown to be effective. For TASC II D (long occlusion) open bypass (OB) is superior to PTA/S in patency and long term outcome. However for TASC II B and C (moderate) lesions there are conflicting reports on the best treatment modality. ROBUST is the first randomized clinical trial comparing OB and PTA/S in TASC II B and C lesions.

Primary objective
Patency (<50% restenosis on duplex), re-intervention rate.

Secondary objectives
Quality of life improvement, morbidity and mortality rate.

Methods:/Study Population
Patients with intermittent claudication who fail medical management or patients with critical limb ischemia are eligible. Patients are maximized medically. Patients with TASC II B or C lesions are prospectively randomized into either OB or PTA/S group; patients with TASC A and D lesion are non-randomized receiving PTA/Stent or Bypass respectively. Patients are followed at 1, 6 and 12 month and yearly up to five years with physical exam, ABI, duplex and QOL questionnaire.

Study Status
Study approved by Hopkins IRB. The study is actively enrolling participants, and making steady progress. Last DSMB Approval on 10/31/12.

ClinicalTrials.gov
Identifier

NCT01602159

Results
Total Consented = 29, Total Enrolled = 22, Screen fail = 7, Withdrawn = 0, Death = 1, TASC A = 4, TASC B = 7, TASC C = 1, TASC D = 10, Total randomized = 8

Patency
One patient with TASC A (non-randomized PTA/S group) and one TASC B (randomized to OB) had restenosis. Both restenosis were treated successfully with angioplasty.

Mortality
No intra operative death reported. One unrelated death at 6-month (TASC B lesion randomized to PTA/S)

Discussion/Significance of Impact
Providing level-1 evidence, ROBUST will establish treatment guidelines in such lesions and eliminate un-necessary procedures and reduce health care costs
Abstract 290: Microrna-199a and Glut1 are Differentially Expressed in Primary Human Arterial and Venous Endothelium and are Deregulated in Culture
Authors: Richardson Matthew R , Lai Xianyin , Yoder Mervin C 
Published: July 2021

Introduction
Studies over the last decade have established that the identity of arterial and venous endothelial cells (ECs) is governed by both genetic and environmental factors. Although the ephrin/Eph system is known to be a key determinant of that identity, it is unlikely that system is solely responsible for all the differences between arterial and venous ECs. Furthermore, microRNAs are becoming increasingly recognized for their important role in regulating gene expression and cell fate. We investigate here novel potential markers and regulators of arterial EC identity and the effects of environmental cues on these molecules.

Methods

ECs were freed from the basement membrane of human umbilical arteries and veins (HUAECs, HUVECs) (n=6) by enzymatic digestion and purified by flow cytometry (CD31+, CD45-) prior to lysis and quantitative PCR or analysis using HPLC-MS/MS. Protein quantification was performed using a proprietary software package (IdentiQuantXL
TM
). HUVECs and HUAECs were harvested directly from umbilical cords and placed directly into collagen coated 6-well plates and cultured in standard conditions in EGM2.

Results
We found that microRNA-199a levels were 23 fold higher in HUVECs than HUAECs (p=0.0001). One of microRNA-199a’s theoretical downstream targets is GLUT1, a facilitative glucose transporter thought to be responsible for basal glucose uptake. GLUT1 mRNA and protein levels were significantly higher (5.5 and 4.3 fold, respectively) in HUAEC than HUVECs (p=0.00002). Using tissue immunohistochemistry (IHC) of we confirmed that GLUT1 expression is restricted to arterial ECs in human umbilical cord and adult peripheral artery and vein sections. Finally, GLUT1 mRNA levels are 2 fold higher in HUAECs than HUVECs in culture at P2 and levels decrease until P5 when they become equal indicating that culture has a homogenizing effect on endothelial heterogeneity.

Conclusions
MicroRNA-199a levels are higher in HUVECs than HUAECs and GLUT1, its downstream target, is higher in HUAECs both at the mRNA and protein level. Protein expression is restricted to arteries in both the new born umbilical cord and adult peripheral vessels. Importantly, expression in both cell types is downregulated and equilibrated by cell culture.
Abstract 368: Family History of Premature Cardiac Events and Sub Clinical Atherosclerosis in Children and Young Adults: A Systematic Review
Authors: Sinha Pragya , Rana Jamal S , Oni Ebenezer T , Aneni Ehimen C , Blumenthal Roger S , Agatston Arthur A , Blaha Michael J , Blankstein Ron , Santos Raul , Nasir Khurram 
Published: July 2021

BACKGROUND
The association between a positive family history (PFH) of premature cardiovascular disease (PCVD) and atherosclerosis has been explored in numerous studies. In adults, various studies have confirmed a significant positive correlation between a PFH and PCVD. Scant literature however, focuses on young individuals. Nevertheless, it is important to understand the impact that a PFH has in young people because the foundations of atherosclerosis and adverse cardiac behaviors develop in youth. In this paper, we aimed to systematically review the evidence linking a PFH of PCVD to indirect markers of subclinical atherosclerosis.

METHODS
The search was conducted on Medline, Web of Science and Embase. ‘Family history’, ‘children/young adults’ and ‘subclinical atherosclerosis’ were the three main concepts used. Increase in mean carotid IMT (cIMT), endothelial dysfunction and vascular inflammation were used as indirect measures of subclinical atherosclerosis.

RESULTS
1191 articles were identified in the initial search. 24 papers with 5400 participants were included in the final review. There were five cohort studies and nineteen case control studies from twelve countries. Mean cIMT was found to be significantly increased in those with a PFH by eleven of the fourteen papers reviewed. Endothelial dysfunction, measured by flow mediated dilatation (FMD), was found to be significantly increased in five of the seven included studies. The evidence on vascular inflammation was somewhat inconsistent with only ten of the nineteen studies demonstrating significance. The results tend to suggest that an elevated mean cIMT, as well as a greater degree of endothelial dysfunction are seen in children and young adults with a PFH of PCVD. Moreover, these differences exist in asymptomatic children as young as 8-9 years (4 studies) in the absence of any other cardiac risk factor.

DISCUSSION
Individuals with a PFH of PCVD have evidence of subclinical atherosclerosis in their youth demonstrating an accelerated tendency to acquire cardiovascular disease. Some of this risk may be attributable to behavioral risk clustering in families. However, a significant proportion of this elevated risk is related solely to a positive family history and needs attention.
Abstract 373: The Association of 9p21.3 Locus with Coronary Atherosclerosis: A Systematic Review and Meta-Analysis
Authors: Munir Muhammad S , Wang Zhen , Steffen Mark W , Erwin Patricia J , Murad Mohammad H , Kullo Iftikhar J 
Published: July 2021

Background
The association between the 9p21-3 locus and coronary heart disease (CHD) is well known. We performed a systematic review of the literature to assess whether this locus was associated with severity of coronary atherosclerosis.

Data Sources
Multiple relevant electronic databases were searched through September 2012 for English language studies with comparative design.

Data extraction
We tested the association of the 9p21-3 locus: 1) measures of severity of coronary atherosclerosis [number of diseased vessels, Gensini Score, Duke CHD Prognostic Index (Dc.PI)]. 2) angiographic outcomes [change in minimum lumen diameter ([[Unable to Display Character: &#8710;]]MLD) and number of new lesions at follow-up] and 3) key clinical outcomes (all-cause mortality, recurrent myocardial infarction and need for coronary revascularization). Genotypes were classified either as homozygous low risk (Lo) or homozygous high risk (Hi).

Data Synthesis
Relative risks (RR) were pooled using the random effects model. Seventeen distinct data sets that included 15,826 participants were analyzed. Ten studies (n=9807) reported number of diseased vessels at baseline. Hi genotype was associated with a 33.7% increased risk of triple vessel disease (95% CI 1.08-1.65; p=0.01). The Hi genotype was associated 0.27 higher baseline Gensini Score (95% CI 0.04-0.48; p=0.01) in 3787 individuals. The locus was not associated with Dc.PI [RR=0.21 (95% CI -0.14-0.55; p=0.25)]. The high risk genotype did not predict [[Unable to Display Character: &#8710;]]MLD or number of new lesions at follow-up. There was no difference between Hi and Lo genotypes in terms of all-cause mortality [RR=1.13 (95%CI 0.90-1.41;p=0.28)], recurrent myocardial infarction (95%CI 0.92-1.40; p=0.24) or frequency of coronary revascularization [RR=1.101 95% CI (0.78-1.57); p=0.56].

Conclusion
An association between 9p21-3 and greater baseline severity of coronary atherosclerosis was seen in this meta-analysis. However no association was seen between 9p21-3 and adverse angiographic and clinical outcomes which may be due to diminishing genetic risk following dietary modification and statin therapy.
Abstract 395: Lipoprotein Apheresis of Hypercholesterolemic Patients Mediates Vasoprotective Gene Expression in Human Endothelial Cells
Authors: Morawietz Henning , Goettsch Winfried , Brux Melanie , Reimann Manja , Bornstein Stefan R , Julius Ulrich , Ziemssen Tjalf 
Published: July 2021

OBJECTIVE
Hypercholesterolemia is an important risk factor of cardiovascular diseases. Lipoprotein apheresis is an efficient strategy to reduce the serum low-density lipoprotein (LDL)-cholesterol and lipoprotein(a) levels and cardiovascular complications in patients with severe hypercholesterolemia. The underlying molecular mechanisms are not well-understood. In this study, we analyzed the impact of lipoprotein apheresis on gene expression in human endothelial cells.

METHODS
Human endothelial cells were stimulated with serum of hypercholesterolemic patients before and after lipoprotein apheresis. The expression of endothelial lipoprotein receptors, nitric oxide (NO) synthase and adhesion molecules was quantified by real-time PCR and Western blot.

RESULTS
Lipoprotein apheresis reduced the expression of the lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) in endothelial cells. Low-density lipoprotein (LDL) receptor expression remained unchanged. The mRNA expression of the endothelial nitric oxide synthase (eNOS) was increased with serum of hypercholesterolemic patients after lipoprotein apheresis. In contrast, endothelial expression of vascular cell adhesion molecule 1 (VCAM-1) was reduced in response to serum after lipoprotein apheresis.

CONCLUSION
Lipoprotein apheresis reduced the expression of the proatherosclerotic oxLDL receptor LOX-1 and adhesion molecule VCAM-1 and increased the expression of vasoprotective and NO generating eNOS in human endothelial cells in response to serum of hypercholesterolemic patients. These novel molecular mechanisms may account for the antiatherosclerotic and vasoprotective potential of lipoprotein apheresis in patients with hypercholesterolemia.
Abstract 409: Size-dependent Acute Cellular Responses of Unbound Free Fatty Acids
Authors: Lee Eun-Soo , Lee Mi Kyoung , Kim Se-hwa 
Published: July 2021

Introduction
The level of free fatty acids (FFAs) in serum determines the pathophysiological condition. Most FFAs are carried in albumin-bound forms, but very small portion is remained unbound. Due to the methodological limitations, there is little known about the effect of unbound form.

Objectives:
This study aimed to understand the effect of local increase of unbound FFAs.

Methods and Results:
We synthesized the nano- and micro-particles of linoleate (LNP and LMP respectively) in unbound forms. The sizes of linoleate particles in distilled water, in serum-free, and in serum-added culture media were measured by dynamic light scattering. We confirmed that the size of LNPs in serum-added media were ~ 200 nm in diameter without aggregation. By the treatments of LNPs and LMPs in macrophage cell line (RAW 264.7), we studied size-dependent cellular responses including viability, lipid storage, and intracellular translocation. Increased lipid droplets in LNPs-treated cells were analyzed based on label-free coherent anti-Stokes Raman scattering imaging. We also found that LNPs (250 μM), not LMPs, activated the translocation toward nucleus of fatty acid binding protein 4 (FABP4), a carrier of intracellular FFA, by western blotting and immunocytochemistry. It was noteworthy that the nano-sized unbound FFA could induce post-transcriptional gene expression unlikely micro-sized FFAs. In the cells whose FABP4 function was inhibited by the chemical inhibitor, the cell death was significantly increased through apoptosis in short exposed time. This cell death response was mediated by endoplasmic reticulum stress suggesting the protective role of FABP4 on the imbalanced FFA metabolisms.

Conclusions:
We showed the size-dependent cellular responses by nano- and micro-sized unbound FFAs and the protective role of FABP4 on the FFA metabolisms. These findings provide the insights into acute cellular responses of unbound FFA.
Abstract 411: Vasculoprotective Function of HDL from Cetp-deficient Subjects
Authors: Ossoli Alice F , Gomaraschi Monica , Pozzi Silvia , Nilsson Peter , Jessup Wendy , Kuivenhoven Jan Albert , Cefalù Angelo , Averna Maurizio , Sirtori Cesare R , Franceschini Guido , Calabresi Laura 
Published: July 2021

Objective-
Cholesteryl ester transfer protein (CETP) is a plasma glycoprotein that catalyses the transfer of cholesteryl esters from HDL to the other plasma lipoproteins. Genetic deficiency of CETP is one of the known causes of primary hyperalphalipoproteinemia and represents a unique tool to evaluate how structural HDL alterations impact on HDL atheroprotective activity. Aim of the present study was to assess the vasculoprotective activity of HDL isolated from carriers of genetic CETP deficiency.

Subjects and Methods-
HDL and HDL subfractions were isolated from carriers of the R37X, Q165X and IVS7+1 CETP mutations and tested for their protein/lipid composition and their anti-inflammatory and NO-promoting activity in endothelial cells.

Results-
HDL and HDL3 from carriers proved to be as effective as control HDL in down-regulating cytokine-induced VCAM-1 expression and in enhancing eNOS expression in endothelial cells. Interestingly, carriers’ HDL2 were significantly more efficient than HDL2 from controls in inhibiting VCAM-1 and enhancing eNOS expression, likely due to their peculiar lipid composition and their higher content of apoE. On the contrary, carriers’ HDL and HDL subfractions were less effective than lipoproteins from controls in stimulating eNOS activation by phosphorylation and NO production, likely because of a reduced content of S1P in carriers’ HDL.

Conclusions-
Large, apoE-enriched HDL that accumulate in genetic CETP deficiency retain their ability to preserve endothelial cell homeostasis, supporting the use of pharmacological CETP inhibition to increase HDL levels and enhance HDL-mediated atheroprotection.