bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 237: MicroRNA patterns in Abdominal Aortic Aneurysm Appear to Reflect Adventitial Inflammation
Authors: Jones Gregory T , Phillips Victoria L , van Rij Andre A 
Published: July 2021

MicroRNAs (miRNA) have been shown to play an important role in vascular biology. In this study we examined the miRNA profile within human abdominal aortic tissue and plasma to identify differential expression patterns between aneurysm cases and controls.
Whole miRNome profiles were assessed using array-based assays (Affymetrix miRNA 2.0) in 27 AAA and 14 control aortic biopsies. Results were validated in 89 AAA and 21 controls using real-time PCR (Taqman-based) assays. A subset of case and control aortic samples were also microdissected. Separate intima/medial and adventitial miRNome profiles were compared with matching total wall. In addition, whole miRNome profiles of EDTA plasma from 22 AAA and 20 age and gender matched controls were examined and validated by RT-PCR in 62 AAA and 62 control plasmas.
In aortic tissue, 18 miRNA were highly significantly (p<1x10-4, q<1x10-3) differentially expressed in AAA versus controls. Variable principal component analysis indicated that these miRNA were expressed in three distinct clusters. MiRNA within each cluster were subsequently validated by RT-PCR. The strongest validated associations were miR-146a (p<0.0004), mir-150 (p<0.001), miR-223 (p<0.0001), and mir-140-3p (p=0.006). Aortic wall sub-layer analysis indicated that total wall miRNA profiles reflected adventitial related miRNA in AAA and intima/medial related miRNA in controls.
In plasma, 12 miRNA were initially identified (including three which were also observed in tissue) however only miR-151-5p (p=0.005) was successfully validated by RT-PCR.
The general pattern of AAA associated miRNA expression indicated regulation of genes known to be involved with B-cell and T-cell function. This was also consistent with the concentration of these miRNA within the AAA adventitia, as these cells are key regulators of the vascular (adventitial) inflammation associated with AAA.
In conclusion, microRNAs appear to have significant potential as AAA biomarkers. However, further work is required to determine the specificity of such associations to account for potential confounders, including as concurrent arterial disease.
Abstract 240: Serum Cholesterol and Angiotensin-Induced Abdominal Aortic Aneurysm in Hypercholesterolemic Mice
Authors: Prins Petra A , Hill Michael , Airey David , Nwosu Sam , Perati Prudhvidhar R , Abramo Jason M , Kon Valentina , Fazio Sergio , Sampson Uchechukwu 
Published: July 2021

Background

Although hyperlipidemia is known to augment the incidence of abdominal aortic aneurysms (AAA) in the AngII-induced model of apolipoprotein E
-/-
mice, its relationship to AAA size is unknown. Therefore, we evaluated the relationship between total cholesterol concentration (TC) and change (delta) in aortic diameter.

Methods
TC was measured in 36 male mice that underwent a 4-week infusion period with saline (n=9) or AngII (1500 ng/kg/min; n=27), along with serial measurements of pulse rate (PR), and pulse (PP), mean arterial (MAP), systolic (SBP) and diastolic (DBP) pressure. A linear mixed effect model was used to assess the relationship between all hemodynamic parameters and delta. Nonparametric and linear regression methods were used to evaluate TC in relation to delta.

Results
TC did not differ between AngII and control mice (Figure, bottom left) (p=0.18). The burden of atherosclerosis was greater among AngII-exposed mice versus control, but did not differ by presence or size of AAA (Figure, bottom right). None of the hemodynamic parameters were predictive of delta (SBP, p = 0.66; DBP, p = 0.66; MAP, p = 0.55; PP, p = 0.66; and PR, p = 0.39). Mean TC was higher among mice with large versus small AAA (552.6 vs. 393.5 mg/ ml, p<0.05; Figure, top right). The nonparametric smoothing line (Figure, top left) suggests a first order relationship between delta and TC (p for trend < 0.001). AngII (ß = 0.48, p < 0.001) and TC (ß = 0.0015, p = 0.003) were independent predictors in the linear model for delta.

Conclusions
Our findings suggest that TC is incrementally associated with AAA size. These findings may have potential clinical relevance for risk assessment in AAA patients.
Figure
Abstract 254: TSP-2 siRNA Coating of Prosthetic Arterial Graft Material to Reduce Intimal Hyperplasia in a Rat Model
Authors: Pratt Wande B , Contreras Mauricio A , Pradhan-Nabzdyk Leena , LoGerfo Frank W 
Published: July 2021

Introduction
Intimal hyperplasia represents a complex pathologic vascular response to injury. Our group previously revealed that TSP-2 was associated with this response, and demonstrated that intraluminal delivery of TSP-2 siRNA can suppress the intimal hyperplastic response in an animal model. Furthermore, we have shown that siRNA can be incorporated into prosthetic graft material using a dip-coating technique. We sought to determine whether this method could be employed for siRNA delivery in order to attenuate the response to arterial injury, and to reduce neointima formation in an in vivo model.

Methods
Carotid artery endothelial cell denudation was performed on male Wistar rats using a 2 Fr Fogarty balloon catheter. ePET graft material was dip-coated in several siRNA solutions (i.e. saline, TSP-2 siRNA, PEI-complexed TSP-2 siRNA) for 50 minutes. The graft was cut longitudinally, and then wrapped circumferentially around the denuded carotid artery. Arterial segments were then isolated at 21 days. TSP-2 gene expression was measured by qRT-PCR, and intimal hyperplasia was assessed by quantitative morphometry.

Results
Balloon denudation increased TSP-2 gene expression by 10-fold, when compared to non-denuded arteries after 21 days (p = .01). Fluorophore-conjugated siRNA confirmed transmural delivery following denudation. When compared to saline treated animals, TSP-2 siRNA reduced TSP-2 mRNA expression by 70% (p = .001). The addition of the transfection reagent PEI to TSP-2 siRNA treated grafts, however, did not significantly augment gene silencing. Morphometric analysis of arterial cross sections demonstrated significantly less neointima formation–as measured by intima-media (I/M) total area ratio–among grafts dip-coated with unmodified and PEI-treated TSP-2 siRNA (I/M: 0.2 and 0.4, respectively) compared to saline grafts (I/M: 1.0, p < .001).

Conclusions
In the rat carotid artery balloon injury model, dip-coating of prosthetic graft material with subsequent perivascular delivery of TSP-2 siRNA is an effective method to achieve transmural transfection and consistent gene silencing, while significantly suppressing the intimal hyperplastic response following arterial injury.
Abstract 284: microRNAs are Novel Plasma Biomarkers for Diagnosis and Prognosis of Abdominal Aortic Aneurysm Disease
Authors: Chernogubova Ekaterina , Eken Suzanne M , Jin Hong , Roy Joy , Hamsten Anders , Eriksson Per , Tsao Philip S , Maegdefessel Lars 
Published: July 2021

MicroRNAs (miRNAs) have been identified as transcriptional and posttranscriptional inhibitors of gene expression, thought to “fine tune” the translational output of their target mRNAs. Recently, they have received much attention regarding their suitability as biomarkers for disease. Our goal was to explore the diagnostic and prognostic value of miRNAs in abdominal aortic aneurysms (AAAs), a disease for which currently no established biomarker exists.
Using a PCR-based array platform, we profiled the 168 most abundant blood miRNAs in 20 patient plasma samples with AAA disease, undergoing surgical repair of their enlarged aorta vs. 20 samples from an age, risk factor, and medication matched control group without aneurysm. We were able to identify a total number of 12 miRNAs being significantly altered in diseased patient samples as compared to controls. We further investigated these 12 miRNAs in plasma (as well as in aortic tissue) from apoE-/- mice with angiotensinII (AngII)-infusion induced AAAs, enabling us to discover a potential prognostic value of miRNAs being released into circulation. Indeed we were able to detect that the expression of 4 out of the 12 miRNAs (miRs-126 and -668 both increased; miRs-24 and -210 both decreased), was substantially modified in plasma samples drawn from mice with AAA immediately before rupture occurred between days 10 and 14 after AngII pump implantation compared to mice with AAA that did not rupture for the remainder of study (28 days), as well as saline-infused controls. Importantly, the expression of miRs-24 and -126 appeared also significantly different in plasma samples from patients with ruptured AAAs (n=7) compared to patients with non-ruptured AAAs (abdominal aortic diameter between 55-78 mm; n=7) and un-diseased controls (n=7).
The present study explores the diagnostic and prognostic biomarker potential of miRNAs being released into circulation during initiation, propagation, and ultimately rupture of AAA disease in mice and humans. The identification of miRs-24, -126, -210, and -663 potentially offers great prognostic value to determine which patients present with an increased risk of AAA rupture.
Abstract 293: Embryological Origin of Human Smooth Muscle Cells Influences Their Ability to Support Vasculogenesis.
Authors: Low Lucinda , Cheung Christine , Bennett Martin R , Sinha Sanjay 
Published: July 2021

Lineage-tracking studies in avian and mouse embryos have revealed that SMCs in different regions of the vasculature are derived from different embryological origins. We have developed an in vitro protocol to differentiate human embryonic stem cells (hESCs) into SMCs through different embryonic lineages, namely neuroectoderm, lateral plate mesoderm (LM) and paraxial mesoderm. As mural cells are thought to augment the formation of functional blood vessels during revascularisation, this study aims to determine whether embryologically-distinct SMCs differ in their ability to support vasculogenesis. We assessed the hypothesis that LM-SMCs are superior in supporting endothelial network formation in vitro.
Using an mStrawberry-expressing hESC line, we derived the three origin-specific SMCs and co-cultured them with GFP-expressing HUVECs in a 3D in vitro matrigel assay. Endothelial network formation was assessed using real-time confocal imaging. Quantitative analysis revealed that LM-SMCs alone had a supportive effect on network formation and survival, with an increase in HUVEC network area after 4 days (3.23-fold increase ± 1.04-fold vs HUVECs alone; p<0.05; n=3) and 8 days (6.38-fold increase ± 0.39-fold vs HUVECs alone; p<0.001; n=3). In addition, LM-SMCs appear to facilitate more complex endothelial networks, with narrower cords and more branch points, compared with the other SMC types and HUVECs alone. To identify whether the LM-SMC-specific influence on network formation and survival was a paracrine effect, the three SMC types were tested in a paracrine 3D in vitro assay, where they shared media with HUVECs seeded in an adjacent well. Preliminary results reveal that the supportive effect of LM-SMCs is, at least partly, paracrine, with an increase in HUVEC network area after 4 days (7.08-fold increase vs HUVECs alone; n=1) and 8 days (21.39-fold increase vs HUVECs alone; n=1). Currently, we are exploring possible soluble factors that may be responsible and investigating this effect in an in vivo matrigel assay. In conclusion, the embryological origin of SMCs influences their functional ability to support vasculogenesis. This research will provide insight into which SMC type will be most effective in future revascularisation therapy.
Abstract 308: Papaverine Inhibits Force of Human Saphenous Vein Through Actin Depolymerization and Myosin Light Chain Dephosphorylation
Authors: Hocking Kyle M , Brophy Colleen M , Putumbaka Gowthami , Komalavilas Padmini 
Published: July 2021

Background
Papaverine has been used to combat vasospasm in human saphenous veins (HSV) as an alternative to high pressure distension techniques. Papaverine acts as a nonspecific inhibitor of phosphodiesterases, increasing both cGMP and cAMP. We hypothesized that the mechanism by which papaverine reduces force through regulation of actin depolymerization and myosin light chain dephosphorylation.

Methods

HSV was pretreated with 1 mM papaverine followed by 5 μM norepinephrine. Force and intracellular [Ca
2+
]
i
levels were concurrently measured using a Fluoroplex. Tissue was snap frozen for biochemical analyses.

Results

Papaverine completely inhibited norepinephrine-induced force development and intracellular [Ca
2+
]
i
. Papaverine pretreatment led to increases in the phosphorylation of heat shock-related protein 20 (HSPB6), dephosphorylation of myosin light chain, and depolymerization of actin. Pretreament of papaverine led to an association of HSPB6 with 14-3-3 eta as determined by immunoprecipitation and mass spectrometry analysis. The interaction of HSPB6 with 14-3-3 leads to dissociation and activation of cofilin, an actin depolymerizing protein.

Conclusions

These results suggest that papaverine-induced force inhibition of HSV involves decreases in intracellular [Ca
2+
]
i
by inhibiting the calcium regulatory pathways, leading to the dephosphorylation of myosin light chain. Additionally papaverine induces the phosphorylation of HSPB6, which leads to depolymerization of actin. Papaverine inhibits HSV spasm via multiple molecular mechanisms.
Abstract 319: Protein Kinase C-delta Regulates Pro-inflammatory Chemokine Expression Through the NF?B Pathway in Vascular Smooth Muscle Cells
Authors: Ren Jun , Wang Qiwei , Morgan Stephanie , Ravichander Aarthi , Singh Ravneet , Liu Bo 
Published: July 2021

Objective-
Inflammation is a key contributor to a plethora of vascular diseases including atherosclerosis and aneurysm. Pro-inflammatory chemokines released by vascular smooth muscle cells (SMCs) play a critical role in vascular inflammation. Previously, we showed protein kinase C-delta (PKCδ) was upregulated in SMCs of inflamed arteries. PKCδ-/- mice are resistant to inflammation as well as apoptosis in models of abdominal aortic aneurysm. We hypothesize that PKCδ modulates the inflammatory response in part through regulation of pro-inflammatory chemokine expression in SMCs.

Method and Results-
To identify PKCδ-regulated chemokines, we used microarray to compare transcription profiles of SMCs infected with AdPKCδ or AdNull. Results showed that 45 out of 203 differentially expressed genes have known inflammatory response roles. RT-PCR analyses confirmed the regulatory effect of PKCδ on selected inflammatory chemokines, including MCP1/CCL2, CCL7, CXCL16 and CX3CL1. Using a mouse aneurysm model, we showed all of these chemokines were elevated in aortae of wildtype, but not PKCδ-/- mice. We postulate that PKCδ regulates chemokine expression via the NFκB pathway. Using MCP-1 as a prototype, a detailed analysis showed PKCδ induced MCP1 mRNA expression (4.25±0.57 fold, n=5, p<0.05) and protein secretion (4.41±0.21 fold, n=3, p<0.05). Analysis of pre-mRNA and mRNA indicated that PKCδ stimulated MCP1 expression at the transcription level rather than by controlling its mRNA stability. Using a NFκB inhibitor (Andrographolide) or siRNA knockdown of NFκB subunit p65 eliminated PKCδ’s effect on MCP1 expression. Physical interaction between PKCδ and p65 was shown in cultured SMCs and aneurysmal aortae by the in situ proximity ligation assay. PKCδ/p65 complexes were detected largely outside the nucleus, indicating the two pathways intersect in the cytosol. This notion is supported by the lack of effect of PKCδ translocation peptides on MCP1. Furthermore, we demonstrated that PKCδ enhanced Ser536 phosphorylation and DNA-binding affinity of p65.

Conclusion-
Our results show PKCδ plays a key role in inflammation through the NFκB-mediated chemokine expression pathway in vascular SMCs and may serve as a suitable target for novel anti-inflammatory therapy.
Abstract 342: Coagulation Pathways Drive Substantial Leukocyte Recruitment Relative to Solitary TLR4-mediated Pathways
Authors: Moerland Matthijs , Naaborgh Lisanne , Kluft Cornelis , Burggraaf Jacobus , Malone Karen 
Published: July 2021

Recruitment of leukocytes is critical for controlling infections. However, chronic leukocyte activity is deleterious in a variety of pathological conditions. As such, leukocyte behavior is an attractive pharmacological target, although knowledge is lacking on the exact triggers driving leukocyte migration. We explored the contribution of coagulation pathways versus TLR4 signaling on the leukocyte recruitment by mimicking non-sterile and sterile leukocyte activation. A sterile inflammatory milieu was generated by spontaneous ex-vivo coagulation of human whole blood, a non-sterile milieu by incubating human whole blood with LPS (at the maximal effective dose), inducing TLR-4 mediated cytokines. The chemoattractant activity of supernatant obtained from these conditions was compared by assessment of primary human leukocyte recruitment in transwell cultures, microscopically and by flow cytometry. Human recombinant IL-8 (100 ng/mL) was used as control chemoattractant.
Ex-vivo coagulation resulted in a strong release of IL-8 (119±58 ng/mL), being 5-fold higher than LPS-induced IL-8 release. Leukocyte recruitment (mainly identified as granulocytes) towards the sterile milieu was greater than TLR4-driven chemoattraction. Comparison of recombinant IL-8-induced and coagulation-induced chemotaxis indicated that chemotaxis towards the coagulated milieu was driven by more factors than IL-8 alone. Additional experiments indicated that fibrinogen cleavage product fibrinopeptide B may be involved, as this dose-dependently increased leukocyte migration.
Coagulation activation drives substantial neutrophil recruitment, primarily mediated by IL-8. For this ex-vivo coagulation model, a stronger and differently mediated leukocyte response was observed compared to TLR4-driven chemoattraction. The model can be used to study the involvement of extracellular coagulation pathways in specific inflammatory pathways that differ distinctly from pathways involved in non-sterile inflammatory conditions. This model, when further optimized, may be helpful as a methodological tool for early evaluation of new treatment modalities for chronic inflammatory diseases.
Abstract 360: Cardiometabolic Effects of Young Adult Overweight and Obesity
Authors: Bernstein Stephen A 
Published: July 2021

INTRODUCTION
Little data exists referencing the cardiometabolic effects in young adult. BMI does not account well for body shape and size and has weaknesses. Waist:height ratio along with other modalities have been assessed for improving health metrics for patient care with balancing strength and utility problems with ease of use.

METHODS
Baseline and sequential anthropometric data was collected from 42 deployed overweight Army soldiers command directed to battalion medical for weight loss while in the combat zone. This data is presented for discussion and correlation with their individual success/failure to lose weight while being supervised to pass the Army’s calculated percentage body fat standards.

RESULTS
Baseline mean waist circumference was 96cm for females, 106cm for males. Initial mean glucose was 102mg/dl for females, 106mg/dl for males. Initial mean HDL was 52 mg/dl for females, 37 mg/dl. Total cholesterol and triglyceride levels were normal for 75%. With intervention, fasting glucose normalized. All began with waist circumference over 55% of their waist:height ratio. Those who brought their waist:height below 55% passed Army standards. Internal motivation was key.

DISCUSSION
Often young adults do not seek healthcare, do not carry health insurance, and do not seek preventive health. Adolescence issues carry into adulthood and with the current obesity trends, only points to more severe and earlier issues with cardiovascular diseases. This group already had metabolic or pre-metabolic syndrome, highlighting the cardiometabolic hazards present. No comparative data exists–this age has largely been ignored/unaddressed. As opposed to BMI, measuring waist circumference is simple, easy, office based measurement. Waist circumference gives a better marker of central adiposity, which correlates to cardiometabolic risk. Adding this measurement helps refine meaning to BMI. Athletes have higher BMIs but normal waist:height ratios and are more normal. Likewise, the ratio may improve defining metabolic syndrome criteria, where 55% should be the trigger. Measuring waist circumference is easy in the office and patient friendly at home–providing a better target and guidance for helping patients improve their overall heart health.
Abstract 362: Characterization of Triglyceride-Rich Remnant Lipoproteins in the Hypertriglyceridemic Fredrickson-Levy Dyslipidemic Phenotypes without Chylomicrons: The Very Large Database of Lipids (VLDL-6)
Authors: Hassan Kamran , Jones Steven R , Martin Seth S , Joshi Parag H , Blaha Michael J , Elshazly Mohamed , Kulkarni Krishnaji R , Toth Peter P 
Published: July 2021

Background
The major dyslipidemia (DL) classes are relatively rare with limited phenotypic characterization across large populations.

Methods

We studied 1,340,614 U.S. adults from the Very Large Database of Lipids. Lipid testing was by ultracentrifugation (VAP, Atherotech, Birmingham, AL). Distributions of total cholesterol (TC), HDL-C, LDL-C and triglycerides (TG) in the database were superimposable with those in the NHANES 2007-2008 survey. We used the following criteria to classify the excess lipoprotein components of DL in subjects with TG >130 mg/dL without chylomicrons (CM): VLDL-C or LDLr-C (directly measured LDL-C - Lp(a)-C -IDL-C) > 90
th
percentile, Type III DL remnant lipoprotein cholesterol (RLP-C = VLDL3-C + IDL-C) excess by criterion of VLDL-C/TG > 0.3. The remaining subjects (continuum) not classified by DL criteria were further stratified by TG/HDL-C <3 vs. >3, associated with insulin resistance.

Results
VLDL3-C/ IDL-C ratio was approximately 1 across DL and the continuum groups. The highest concentration of RLP-C was in Type III DL; lesser RLP-C elevation is present in all DL characterized by VLDL excess. The highest non-HDL-C was present in type IIb with lesser elevations in types III and IV. RLP-C was higher in the TG/HDL-C>3 continuum than TG/HDL-C<3 continuum reference group (Table).

Summary
In addition to type III, the major non-CM DL is frequently associated with excess RLP-C. RLP-C tended to be normal in the continuum subjects, trending higher in those with TG/HDL-C>3.