bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 492: Arachidonic Acid Induces Activation of Platelet PF4 and Par-1 mRNA, Which Is Attenuated by Aspirin
Authors: Hu Liang , Nardi Michael A , Merolla Michael , Suarez Yajaira , Berger Jeffrey 
Published: July 2021

Arachidonic acid (AA) is converted to thromboxane A2 via the cyclooxygenase pathway; however its exact mechanism of platelet activation is uncertain. Inhibition of this pathway via aspirin highlights the importance of this pathway in decreasing thrombotic events.
In the present study, we investigate the effect of AA on platelet activity indicators (leukocyte- and monocyte-platelet aggregation [LPA, MPA] and reticulated platelets [RP]), as well as the expression (mRNA and protein) of platelet markers PF4 and Par-1, previously well established platelet transcripts with quantitative determinations. To this end, whole blood was incubated with AA (150mM) for 30 min at room temperature in the absence or presence of aspirin (1mM) prior to addition of antibodies for platelet activity indicators, and isolating platelets for mRNA and protein expression.
LPA and MPA were significantly increased after AA stimulation in a dose dependent manner, and were inhibited by aspirin treatment. AA significantly increased PF4 and Par-1 protein level as determined by flow cytometry and western blot assays. Pretreatment with aspirin also attenuated this increase in protein levels. Surprisingly, AA stimulation significantly increased thiazole orange staining (a measure of nucleic acids), another marker of increased platelet activity. Importantly, these results suggest that AA-mediated platelet activation produced an overall increase in platelet total RNA content. To confirm these findings, we analyzed the mRNA expression of PF4 and Par-1 by quantitative real time PCR from platelets treated with AA. Interestingly, AA significantly up-regulated the platelet mRNA transcripts of PF4 and Par-1 by 40% to 60%, and pretreatment with aspirin completely attenuated this effect supporting the specificity of the AA effect on platelet RNA.
Altogether, these data suggest that platelet mRNA is affected by AA stimulation, which is attenuated by pretreatment with aspirin. However, the mechanisms responsible for the increased mRNA levels and expression of PF4 and Par-1 (processing of pre-RNA to mRNA) require further investigation. Importantly, our findings provide novel insight regarding platelet activation and a better understanding of mediators in the processes of thrombosis and hemostasis.
Abstract 1: Molecular Basis of Hypertriglyceridemia Associated with the APOA5 c553.G>T SNP
Authors: Sharma Vineeta , Forte Trudy M , Ryan Robert O 
Published: July 2021

Objectives
Apolipoprotein (apo) A-V is a low abundance protein with profound effects on plasma triglyceride (TG) levels. Several APOA5 SNPs correlate with hypertriglyceridemia (HTG). The c.553 G>T SNP, substituting a Cys for Gly at position 162 of mature apoA-V, is prevalent in Asian populations and correlates with HTG. To investigate the mechanism underlying this association, gene transfer studies were performed in apoa5-/- mice.

Methods
Adeno-associated virus (AAV2/8) harboring the coding sequence for wild type apoA-V (AAV2/8-apoA-V), G162C apoA-V (AAV2/8-G162C) and LacZ (AAV2/8-LacZ) were injected (1x10e12 virus genome) into the tail vein of 8 apoa5-/- mice per group. Blood samples were collected weekly for 4 weeks and TG and apoA-V levels measured. FPLC was performed on plasma obtained from AAV2/8-apoA-V and AAV2/8-G162C mice. VLDL, LDL, HDL and the lipoprotein-free region were characterized.

Results
Compared to AAV2/8-LacZ mice, AAV2/8-apoA-V mice had significantly lower plasma TG levels (50% ±5). Unlike AAV2/8-apoA-V, mice injected with AAV2/8-G162C displayed little or no reduction in TG despite similar amounts of plasma apoA-V protein. Immunoblot analysis of FPLC fractionated plasma revealed that, whereas wild-type apoA-V was lipoprotein associated (VLDL and HDL), G162C apoA-V was largely recovered in the lipoprotein-free fraction. Immunoblot analysis following SDS-PAGE under non-reducing conditions revealed that lipoprotein-associated wild type apoA-V and G162C apoA-V are monomeric; by contrast, the electrophoretic mobility of G162C apoA-V recovered in the lipoprotein-free fraction was retarded.

Conclusions
Gene transfer of wild type apoA-V induces a significant reduction in plasma TG levels of apoa5-/- mice. By contrast, G162C apoA-V failed to induce a corresponding decrease in plasma TG, recapitulating effects observed in human populations harboring this SNP. The propensity of G162C apoA-V to form a disulfide bond with one or more plasma proteins interferes with its lipoprotein binding ability, resulting in loss of function. The results provide a molecular explanation for HTG associated with a common APOA5 SNP. The gene transfer strategy employed provides a platform for studies of other common apoA-V SNPs.
Abstract 125: Weight Loss Selectively Increases High-density Lipoprotein without apoC-III, a Subfraction with Protective Association Against Coronary Heart Disease.
Authors: Wang Liyun , Talayero Beatriz G , Furtado Jeremy D , Bray George A , Sacks Frank M 
Published: July 2021

Human high-density lipoprotein (HDL) particles are highly heterogeneous in composition, and in relation to coronary heart disease (CHD). Specifically, HDL containing apoC-III predicts
increased
incidence of CHD, whereas HDL that does not contain apoC-III predicts
decreased
CHD incidence. Recently, we found that obese subjects had a higher concentration of HDL with apoC-III and a lower concentration of HDL without apoC-III than normal weight controls. However, the effects of weight loss on HDL with or without apoC-III are not known.

We studied 39 obese participants (mean BMI of 34) in a weight-loss trial assigned to a high fat 40%/high protein 25%/low carbohydrate 35% diet for 2 years. We chose this diet group because HDL-cholesterol (-C) levels increased the most compared to the other diet groups with more carbohydrate and less fat. Participants with at least 8% loss of initial body weight at 2 years were assigned to the weight loss group (n=21), while those without a change in weight comprised the no-weight loss group (n=18). Subjects were matched by age (mean 51 years), gender, and race. HDL without apoC-III and HDL with apoC-III were separated from plasma by sequentially using anti-apoA-I and anti-apoC-III immunoaffinity chromatography.
For both groups, the mean baseline HDL-C and total plasma apoA-I concentrations were 48 and 119mg/dL, respectively. Weight loss increased HDL-C concentration by an average of 25% (p<0.01 for changes between groups), and total apoA-I by 9% (p<0.02). In the weight-loss group, mean apoA-I concentrations of HDL without apoC-III increased by12% (from 99 to 111 mg/dL) compared to a decrease of 6% in the no-weight loss group (from 107 to 101mg/dL) (p<0.02). The apoA-I concentration of HDL with apoC-III, 15 mg/dL, remained unchanged.

In conclusion
, moderate weight loss selectively increased the concentration of HDL without apoC-III, the predominant HDL type that predicts reduced incidence of CHD. In contrast, weight loss did not affect HDL with apoC-III, an HDL type associated with increased CHD risk. This represents a partial normalization of HDL subtypes by moderate weight loss. This effect on HDL subpopulations defined by absence of apoC-III suggests a novel mechanism by which weight loss may reduce cardiovascular disease risk.
Abstract 159: Comparing Blood Pressure Control in Diabetic vs Non-Diabetic Veterans following Percutaneous Coronary Intervention
Authors: Blumenkrants Dmitry , Siddiqui Saifullah M , Challa Karthik , Ladani Amit , Shroff Adhir 
Published: July 2021

Background
As per the US Joint National Committee VII (JNC-7) recommendations, patients with known underlying coronary artery disease and diabetes should have goal blood pressures (BP) of systolic (SBP) <130 and diastolic (DBP) <80 to decrease morbidity and mortality associated with cardiovascular disease. In addition to lifestyle modification, these goals can be attained by use of multiple classes of drugs including beta-blockers (BB), angiotensin agonists (ACE-I/ARB), calcium channel blockers (CCB), diuretics and nitrates.

Methods
We conducted a retrospective cohort study focusing on the attainment of the JNC-7 guidelines, comparing outcomes between 415 diabetic to 637 non-diabetic patients undergoing PCI between September 2004 and December 2012 at the Jesse Brown Veterans Administration Hospital in Chicago, IL. Blood pressure (BP) measurements and antihypertensive medications pre and post PCI at 6-month follow-up were documented.

Results
Among the diabetic population, the mean SBP decreased from 136 to 131 mmHg (p < 0.0001) and mean DBP decreased from 73 to 70 mmHg (p < 0.0001). In the non-diabetics, the mean SBP decreased from 133 to 127 mmHg (p < 0.0001) and the mean DBP decreased from 74 to 71 mmHg (p < 0.0001).
With regards to JNC-7 guidelines, the percent of diabetics at SBP goal increased from 42% to 49% (p = 0.047) and percent at DBP goal increased from 74% to 82% (p = 0.008). In non-diabetics, percent at goal for SBP increased from 46% to 57% (p < 0.0001) and percent at DBP goal increased from 68% to 76% (p = 0.003). Among diabetics, there was a statistically significant (p <0.0001) increase in use of BB from 77% to 90%. In non-diabetics, there was a statistically significant (p <0.0001) increase in use of BB from 64% to 86% and ACE-I/ARB from 51% to 70%.

Conclusions
In both groups (diabetics and non-diabetics) undergoing PCI, both systolic and diastolic blood pressure improved with more patients achieving JNC-7 targets. Among diabetics, there was a significant increase in utilization of BB. Among non-diabetics, there was a significant increase in utilization of BB and ACE-I/ARB.
Abstract 160: The Role of Inhibitor of Differentiation 3 in the Regulation of Monocyte Chemoattractant Protein-1 Within Epididymal Adipose Tissue During Early Stages of Obesity
Authors: Kaplan Jennifer L , Garmey James C , Harmon Daniel B , Marshall Melissa A , McNamara Coleen A 
Published: July 2021

Background

Global deletion of helix-loop-helix (HLH) transcription factor Inhibitor of Differentiation 3 (Id3) attenuates HFD-induced obesity. Specifically,

Id3
-/-

mice have less epididymal adipose tissue (eAT); an effect that may be linked to eAT inflammation. Adipose tissue macrophages play a key role in HFD-induced obesity and are recruited by Monocyte Chemoattractant Protein-1 (MCP-1). MCP-1 is produced by a wide variety of cells.

Id3
-/-

mice have attenuated HFD-induced levels of MCP-1 mRNA and protein in eAT. Consistent with lower levels of MCP-1 in eAT,

Id3
-/-

mice also have decreased numbers of proinflammatory macrophages in eAT after HFD. Id3 is broadly expressed and involved in many disease pathways. It is currently unclear how Id3 is regulating MCP-1 in eAT, or in which MCP-1-producing cell.

Results

Id3
-/-

mice and
C57Bl/6
littermate controls were fed HFD (60% fat) for 1 and 4 weeks. Stromavascular fraction (SVF) cells from eAT were cultured with Brefeldin A to inhibit protein secretion. Intracellular levels of MCP-1 were analyzed via flow cytometry. Both 1 and 4 weeks of HFD increased the number of MCP-1+ cells, compared to chow-fed controls. There were two levels of MCP-1 expression. Adipocyte progenitors, identified as CD31-CD45-Ter119-CD34+CD29+Sca-1+, express high levels, and macrophages, identified as CD45+F4/80+CD11b+, express mid levels of MCP-1. MCP-1 high cells increased after 1 week of HFD, and this increase was amplified after 4 weeks of HFD. MCP-1 mid cells did not increase until 4 weeks of HFD. However, in

Id3
-/-

mice, both populations were decreased in number at both stages of HFD. MCP-1+ cells in

Id3
-/-

mice also had lower mean fluorescence intensity (MFI) compared to WT controls, indicating lower levels of MCP-1 per cell.

Conclusion
Global deletion of Id3 decreases the number of MCP-1 expressing cells in epididymal adipose tissue, as well as MCP-1 levels per cell. These data suggest that Id3 promotes an immediate response to HFD via an increase in the number of MCP-1 producing cells. There is also evidence that Id3 is directly regulating expression of MCP-1 within adipocyte progenitors. Further studies will elucidate the exact mechanism of regulation of both the number of cells and expression of MCP-1 by Id3.
Abstract 179: Vascular Inflammation and Hepatic Steatosis among the Overweight Metabolically Healthy Individuals and Normal Weight with Cardiometabolic Risk Factor Clustering
Authors: Shaharyar Sameer , Karim Adil , Blaha Michael J , Ali Shozab S , Jamal Omar , Agatston Arthur , Blumenthal Roger S , Nasir Khurram , Santos Raul D 
Published: July 2021

BACKGROUND
Emerging literature has elucidated a new phenotype, metabolically healthy overweight/obese (MHO) who have a similar CVD risk to normal weight individuals. The aim of the study was to compare the relative risk of both phenotypes with subclinical inflammatory-cardiometabolic disorders.

METHODS
We evaluated 6,464 healthy non diabetic Brazilian subjects (43±10 years, 79% males). Those with <2 metabolic risk factors (MRF)were considered metabolically healthy and those with BMI≥25 kg/m2 as overweight (OW). High sensitivity C reactive protein (hs-CRP) was measured to assess degree of underlying inflammation and hepatic steatosis (HS) was determined via abdominal ultrasound.

RESULTS
Overall, 52% OW individuals were metabolically normal, and 11% normal-weight had≥2 MRF. The respective prevalence for elevated CRP (≥ 3 mg/dl) was 28% vs 16%, p<0.0001) and 71% vs 28% for HS, respectively. As compared to those with normal weight/metabolically benign, the highest risk of elevated CRP & HS was noted among those with overweight/metabolically unhealthy individuals (table). Both MHO individuals & normal weight metabolically unhealthy participants had similar association with elevated CRP & HS.

CONCLUSION
Our study highlights that healthy overweight/obesity & normal weight metabolically unhealthy phenotypes are not entirely benign and efforts are needed to reverse these conditions.
Abstract 187: Regulation of Vascular Smooth Muscle Cell (VSMC) Inflammation and Migration by the Pro-resolving Lipid Mediator Maresin-1 (mar1)
Authors: Chatterjee Anuran , Toy Robert , Mottola Giorgio , Chen Mian , Conte Michael S 
Published: July 2021

Introduction
Resolution of acute inflammation is regulated by endogenous lipid mediators derived from polyunsaturated fatty acids such as docosahexaenoic acid (DHA), however little is known about mechanisms of resolution in vascular injury. We investigated the effects of the DHA-derived mediator Mar1 on VSMC phenotype responses.

Methods
Primary human VSMCs were obtained from saphenous vein. VSMC were pretreated with Mar1 (10-100nM) then exposed to TNF-α (10ng/ml), and inflammatory responses assessed using a monocyte adhesion (U937) assay, expression of cell adhesion molecules and pro-inflammatory molecules (qPCR, western blot, ELISA), and production of superoxide (DHE). VSMC migration was measured in a transwell assay with PDGF-AB as the agonist, and cyotskeletal changes were assessed by actin-phalloidin staining.

Results
Mar-1 (100 nM) reduced U937 adhesion to TNF-stimulated VSMC, VCAM-1, and pro-inflammatory cytokine (IL-6, IL-8) expression. Superoxide production measured by DHE fluorescence was reduced by 57% (p=0.002) and Nox4 expression was markedly attenuated (43%, p=0.01). Mar-1 (0.01-100nM) induced rapid cytoskeletal changes with increased cell area, and reduced VSMC migration (76%, p=0.004) to PDGF-AB (50ng/ml; Figure).

Conclusions
Mar-1 attenuates TNF-α inflammatory activation of VSMC, with reduction in pro-inflammatory gene expression, oxidant stress, and monocyte adhesion. Mar-1 reduces actin polymerization and inhibits VSMC chemotaxis to PDGF. Pro-resolving mediators may represent a new class of endogenous vascular therapeutics.
Abstract 197: Emerging Role of Toll-like Receptor 4 and Heat Shock Protein 60&70 in Ischaemia-Induced Skeletal Muscle Damage In Vitro
Authors: Navi Ali , Yu Rebekah , Shi-Wen Xu , Shaw Sidney , Baker Daryll , Tsui Janice 
Published: July 2021

OBJECTIVES
The innate immune response contributes to the skeletal muscle damage in patients with critical limb ischaemia (CLI); however, the detailed signaling mechanisms are not fully understood. We hypothesized that simulated ischaemia induces inflammatory cytokine release from skeletal myotubes, via a mechanism that involves heat shock protein (HSP) 60&70, known endogenous ligands of Toll-like receptor 4 (TLR4), in vitro.

METHODS
Human gastrocnemius muscle biopsies were taken from patients with CLI undergoing major lower limb amputation and from patients with no peripheral arterial disease (PAD). Human myoblasts were isolated, cultured to myotubes and then pre-treated with TLR4 neutralizing antibody prior to exposure to simulated ischaemia. Fluorescent immunostaining was carried out to confirm cell differentiation; ELISA analysis were carried out to quantify IL6 and TNF-α release; and Western blot was used to assess expression of HSP60&70, TLR4 and cleaved caspase-3 as a marker of apoptosis.

RESULTS
Myotubes from patients with CLI expressed greater levels of cleaved caspase-3 and TLR4 as compared to those from patients with no PAD. When exposed to ischaemic conditions, increased IL6 and TNF-α release and upregulation of HSP60&70, cleaved caspase-3 and TLR4 were observed in myotubes from both groups of patients compared to culturing in normoxic conditions (P<0.05). Pre-treatment of myotubes from patients with CLI with TLR4 neutralizing antibody prior to simulated ischaemia was associated with reduced expression of HSP60&70, IL6, TNF-α and cleaved caspase-3 (P<0.05).

CONCLUSIONS
Increased cytokine release, apoptosis and expression of HSP60&70 and TLR4 occur in ischaemic skeletal muscle in vitro. TLR4 antagonism was associated with reduced apoptosis and inflammatory cytokine release and down-regulation of HSP60&70 expression. This suggests a potential pathway where TLR4 and its endogenous ligands contribute to a positive feedback loop to maintain a proinflammatory environment during ischaemia.
Abstract 204: Brilliant Blue FCF is an Alternative Vein Marking Dye That Restores Functional Viability in Human Saphenous Vien Graft
Authors: Brophy Colleen , Hocking Kyle M , Komalavilas Padmini , Lander Cynthia , Cheung-Flynn Joyce 
Published: July 2021

Objective
The human saphenous vein (HSV) is the most effective conduit for infrainguinal peripheral bypass grafting. Optimal vein graft preparation is critical to vein graft patency. Graft marking using surgical skin markers is detrimental to endothelial and smooth muscle function of the graft. The objective of this study was to evaluate the effect of FD&C Brilliant Blue No. 1 (FCF) as an alternative for vein graft marking.

Methods
Segments of HSV were collected after typical harvest and graft preparation. The tissues were cut into rings and either left unmarked or marked with FCF (2.6 mM). The functional viability of the rings was determined in a muscle bath.

Results

Marking with FCF did not impair contractility of the smooth muscle (Figure 1A). Contractility was

restored

in HSV segments that had no functional contractile responses after harvest and preparation (Figure 1B). To determine the mechanism of this protective effect of FCF, purinergic receptors were activated with BzATP with and without pre-incubation with FCF (50μM) or a purinergic receptor antagonist (oATP, control) and the rise in [Ca
2+
]
i
was measured. FCF and oATP reduced the BZATP-evoked rise in [Ca
2+
]
i
(Figure 1C), suggesting that the salutary effects of FCF were due to inhibition of a P2X or P2Y receptor.

Conclusions
FCF is a non-toxic alternative to surgical skin markers that antagonizes purinergic receptors in HSV. FCF restored functional viability to injured vascular smooth muscle. FCF may protect HSV against injury-elicited ‘danger’ signals such as extracellular ATP that is released during injury-induced preparation.
Abstract 206: Small GTPase Rap1 Transmits Mechanical Signals to Control Vascular Tone and Blood Pressure
Authors: Lakshmikanthan Sribalaji , Zheng Xiaodong , Zieba Bartosz J , Nishijima Yoshinori , Ge Zhi-Dong , Momotani Ko , Lund Hayley , Maas Jason E , Szabo Aniko , Vasquez-Vivar Jeannette , Mattson David L , Auchampach John A , Somlyo Avril V , Zhang David X , Chrzanowska-Wodnicka Magdalena 
Published: July 2021

Smooth muscle (SM)-mediated vessel contraction and endothelium-dependent vessel dilation are major mechanisms controlling basal vascular tone. Increased vascular tone leads to arterial hypertension, a risk factor for cardiovascular diseases. Agonist-induced, Ca2+-dependent SM contraction is modulated by intracellular cAMP, which promotes SM relaxation by inhibition of Rho-mediated myosin regulatory light chain (RLC20) phosphorylation and actomyosin contractility. In response to elevated blood flow, endothelium releases dilator substances that relax vascular SM, and in particular defects in production of nitric oxide (NO) lead to hypertension.
Rap1 is a small GTPase that integrates signals from multiple receptors and two Rap1 isoforms, Rap1a and Rap1b are ubiquitously expressed. We found global Rap1b deletion leads to cardiac hypertrophy and hypertension and examined the effect of Rap1-deficiency on vascular tone. Using SM and endothelium tissue-restricted Rap1 knockout mice we show that deletion of Rap1, via distinct mechanisms in the two tissues, contributes to elevated vascular tone. In SM Rap1 regulates basal contraction level and mediates cAMP-induced desensitization of contraction. Rap1 suppresses RhoA-mediated RLC20 phosphorylation, Ca+2 sensitization and relaxation, signaling that is further enhanced upon cAMP-dependent activation of Rap1 by Rap1 GEF Epac.
In endothelium, we find Rap1 is essential for regulation of NO-dependent vasodilation, as deletion of three of the four Rap1 alleles leads to a significant decrease in NO-dependent vasodilation. Significantly, we find that Rap1 is required for normal shear stress-induced NO release and Rap1-deficency in endothelium leads to elevated blood pressure in mice. Mechanistically, we show that Rap1 is required for transducing signals from the endothelial mechanosensing complex triggering signaling leading to NO production. Because shear stress from flowing blood is the main determinant of NO release, this novel finding positions Rap1 as a key regulator of endothelial function.
In conclusion, Rap1 in SM and endothelium, via distinct mechanisms, plays a key role in maintaining normal vascular tone and blood pressure.