bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 628: Circulating Exosomes Isolated From Septic Mice Induce Endothelial Hyperpermeability Through Promoting Podosome Cluster Formation
Authors: Mu Xingjiang , Wang Xiaohong , Essandoh Kobina , Li Yutian , Pugh Amanda M , Peng Jiangtong , Deng Shan , Salem Esam S , Caldwell Charles C , Fan Guo-Chang
Published: July 2021

Introduction:
Septic shock increases vascular permeability, leading to multiple organ failure and higher mortality. Reactive oxygen species (ROS) have been shown to promote both actin cytoskeleton reorganization resulting in vascular leakage and the formation of podosome, an actin-based dynamic membrane structure. Interestingly, recent studies have shown that circulating exosomes from septic patients contained higher levels of ROS than healthy ones. In this study, we hypothesized that septic exosomes can transfer exosomal ROS to endothelial cell (EC) to promote the generation of podosomes, leading to hyperpermeability.

Methods:
C57BL/6 mice (20-25g) were subjected to CLP surgery or injection with LPS (25 μg/g). Sham-operated or PBS-treated mice were used as controls. Exosomes were isolated from sera, collected at 3 h post-CLP or post-LPS-injection. Podosomes were identified by co-immunostaining F-actin/cortactin following stimulation with PMA, thrombin and exosomes. Transendothelial electrical resistance (TEER) analysis was performed to monitor the change of EC monolayer permeability. The ROS levels in exosomes and ECs were measured by using a ROS-Glo H
2
O
2
Assay kit.

Results:
First, we observed that thrombin and PMA both stimulated podosome cluster formation at the cell periphery in (30-36%, 90-108 of 300) ECs, which correlated well with reduction (40.5-50.7%, 6-7.5 of 14.8) in TEER values (n=4, p<0.01). Next, we discovered that septic exosomes collected at 3 h post-CLP or post-LPS-injection contained significantly higher amount of H
2
O
2
which can be transferred into ECs, leading to 1.5-fold increase of H
2
O
2
in ECs (n=6, p<0.01). Moreover, treatment of ECs with septic exosomes remarkably increased the percentage of cells carrying podosome clusters (20.3%, 51 of 251) compared to controls (9.5%, 24 of 252 n=4, p<0.01). Meanwhile, the TEER value was reduced by 27.6% (3.4 of 12.3) upon septic exosome stimulation (n=4, p<0.01). By contrast, scavenging H
2
O
2
in septic exosomes significantly attenuated their effects on podosome cluster formation and EC monolayer TEER.

Conclusion:
septic exosomes can stimulate the formation of podosome clusters in ECs through transferring exosomal H
2
O
2,
leading to endothelial hyperpermeability.
Abstract 629: IFN-Gamma-Dependent Repression of TET2 in Allograft Vasculopathy Exacerbates VSMC Activation
Authors: Ostriker Allison , Martin Kathleen
Published: July 2021

Coronary allograft vasculopathy (CAV) occurs in 50% of heart transplant recipients at 10 years after surgery. Allograft failure secondary to CAV accounts for 30% of deaths in heart transplant recipients. CAV is characterized by concentric intimal hyperplasia (IH) in the graft vasculature that causes ischemic injury to the myocardium. T cell-derived INF-γ is a well-established driver of IH in CAV. The molecular mechanisms of IFN-γ-dependent phenotypic modulation of vascular smooth muscle cells (VSMCs) in CAV are not fully elucidated.

Our group recently showed that TET methylcytosine dioxygenase 2 (TET2) is a master regulator of VSMC phenotype. We hypothesized that IFN-γ modulates TET2 expression and/or function in VSMCs in CAV. We found, using a minor histocompatibility mismatch aorta graft model of allograft vasculopathy, that TET2 expression was decreased in the graft neointima relative to aortic VSMCs prior to grafting. Lineage tracing studies showed that TET2 activity was decreased in donor VSMC-derived cells present in the neointima. Nuclear localization of phospho-STAT1 correlated with TET2 repression in these cells suggesting that IFN-γ signaling may mediate repression of TET2. Further, we found that IFN-y was sufficient to repress TET2 expression in human and mouse VSMCs in vitro. TET2 repression by IFN-γ was not repressed by rapamycin, an mTOR inhibitor known to be effective against CAV but which has significant side effects.

We hypothesized that IFN-γ signaling directly represses TET2. IFN-γ rapidly induced activation of STAT1 in VSMCs in vitro. Transcription factor binding site prediction suggested potential STAT1 binding sites on the TET2 promoter in both human and mouse. ChIP-PCR confirmed that STAT1 occupancy occurs at the TET2 promoter in response to IFN-γ.

Studies are ongoing to determine if STAT1 directly negatively regulates TET2 transcription in response to IFN-γ, and if TET2 overexpression can rescue the effect of IFN-γ on VSMC activation and IH in in vivo models of allograft vasculopathy. Since IFN-y-dependent repression of TET2 is not reversed by rapamycin, elucidating this pathway represents an opportunity for developing complementary therapies to mTOR inhibition that may result in improved outcomes and fewer side effects.
Abstract 630: Cst1 Discriminates Between Senescence Due to Replicative and Stress-Induced Endothelial Cell Aging
Authors: Afreen Sadaf , Herbert Karl E
Published: July 2021

Introduction:
Senescence comprises the cellular and molecular changes leading to compromised functionality of organs. The presence of senescent endothelial cells in human atherosclerotic lesions suggests a contribution to vascular pathology. Better understanding of endothelial senescence will help identify its role in endothelial dysfunction. In this study gene expression changes were assessed in human endothelial cells following induction of senescence either by replicative or peroxide-induced stress and genes identified that were differentially expressed to be used as biomarkers of endothelial aging.

Methods:
Replicative senescent endothelial cells (REPS) were established by passaging human umbilical vein endothelial cells (HUVECs) up to 25 population doublings and stress-induced premature senescence (SIPS) was induced by treating HUVECs with tertiary butyl hydrogen peroxide (t-BHP) for 1 hr for consecutive 3 days. Senescence was confirmed by staining with senescence marker SA β-gal and p53 expression through Western blotting. Gene expression changes were confirmed by qPCR using Taqman probes.

Results:
REPS and SIPS cells were found to be 71 % (912 of 1285) and 81 % (980 of 1210) positive for SA-beta gal staining respectively, compared to young endothelial cells (10 % (130 of 1308); Fig1). CST1 was identified as highly expressed gene (90 fold) in REPS but not SIPS of endothelial cells in microarray gene expression analysis. Further validation using qPCR showed a 164 fold increase in CST1 expression (fig2) in REPS endothelial cells only.

Conclusion:
In conclusion, this study confirms that CST1 as a new marker of replicative senescence in human endothelial cells.
Abstract 631: Nox1-Mediated CREB Promotes Gremlin1-Induced Endothelial Cell (EC) Proliferation
Authors: de Jesus Daniel , Sahoo Sanghamitra , Gorelova Anastasia , Cifuentes-Pagano Maria Eugenia , Pagano Patrick
Published: July 2021

Background:
Pulmonary arterial hypertension (PAH) is a devastating, rapidly degenerating disease characterized by lung vascular cell proliferation/remodeling and elevated vascular pressure leading to right heart failure. We previously showed that BMP antagonist-Gremlin1 elicits pulmonary endothelial cell (EC) proliferation in response to hypoxia, a stimulus that recapitulates in vivo changes occurring in human PAH also leading to vascular cell proliferation and remodeling. NADPH oxidase-derived reactive oxygen species (ROS) purportedly play a critical role in PAH; yet the mechanisms by which they propagate the disease are scant. Other studies show that the cAMP response element protein (CREB) is activated by ROS and modulates cell proliferation. We postulated that Nox1-mediated CREB activation leads to Gremlin1 expression promoting human pulmonary arterial EC (HPAEC) proliferation.

Method:
HPAECs were subjected to 24 hrs hypoxia (1% O
2
) vs. normoxia (21% O
2
). Nox1, Nox2, Nox4, active CREB (pCREB), Gremlin1 and active Smads1/5/8 (pSmads) were evaluated by Western blot. Superoxide anion (O
2
•-
) changes were assessed using cytochrome C. To evaluate whether CREB binds to Gremlin1 promoter, chromatin immunoprecipitation (CHIP) was performed.

Results:
Hypoxia upregulated Nox1 (58% increase vs. normoxia, p<0.0001) but did not affect Nox2 and 4 levels. Hypoxia induced O
2
•-
(13.6±1.5 vs. 18.3±0.6 nmol/min*mg, normoxia v. hypoxia, respectively, p<0.05). Furthermore, hypoxia increased pCREB (66% vs. normoxia, p<0.05), and Gremlin1 (39% increase vs. normoxia) whereas it decreased pSmads 1/5/8 (50% reduction vs. normoxia, p<0.05). Nox1 siRNA decreased pCREB (48% reduction vs. hypoxia alone, p=0.08). Finally, preliminary data show CREB binding to the Gremlin1 promoter.

Discussion:
In the present study, we found that hypoxia-induced HPAEC O
2
•-
derived from Nox1 appears to mediate CREB activation, and subsequent promotion of Gremlin1 expression. Taken together, our data are consistent with CREB mediating Nox1-Gremlin1 signaling in hypoxia-induced EC proliferation.
Abstract 638: Novel Role of 4-Hydroxy-2-Nonenal Mediated Mitochondrial Stress Signaling in Ischemia and Reperfusion Injury
Authors: Miriyala Sumitra , Chandra Mini , Panchatcharam Manikandan , Kevil Christopher , Bhuiyan Md Shenuarin , Fox Jonathan
Published: July 2021

Myocardial infarction is the single most prevalent cause of morbidity and mortality among adults. Excess generation of reactive oxygen species plays a major role in the cellular response to cardiac ischemia/reperfusion (I/R) injury. Accumulated evidence indicates that oxidative stress in mitochondria plays an important role in I/R injury, but how mitochondrial redox mechanisms are involved in cardiac dysfunction remains unclear. Manganese Superoxide Dismutase (MnSOD), an antioxidant enzyme that catalyzes the conversion of superoxide radicals (O
2
•-) in mitochondria. The absence of
SOD2
(a gene that encodes MnSOD) is found to be embryonic lethal in animal models due to impairment of mitochondrial function, most noticeably in the heart. In our investigation, we found MnSOD mimetic, MnTnBuOE-2-PyP
5+
distributed 3-fold more in mitochondria than in cytosol. The exceptional ability of MnTnBuOE-2-PyP
5+
to dismute O
2
•- parallels its ability to reduce ONOO– and CO3–. Based on our initial results, we have generated mice that specifically lack MnSOD in cardiomyocytes (Mhy6-SOD2
Δ
). These mice showed early mortality ~6 months due to cardiac mitochondrial dysfunction. FACS analyses using Mito-Tracker Green indicated that the mass of mitochondria per cell was slightly decreased in the Mhy6-SOD2
Δ
to the wild type. We then examined oxidative phosphorylation levels in Mhy6-SOD2
Δ
v.s. wild type using a Seahorse XF analyzer. The rate of oxygen consumption per cells was significantly lower in Mhy6-SOD2
Δ
cardiomyocytes than that in wild type. The most noticeable difference in the O
2
consumption was found in the presence of FCCP (H+ ionophore/uncoupler). 4-hydroxy-2-nonenal (HNE) adduction of mitochondrial apoptosis-inducing factor (AIFm2) inactivates the NADH oxidoreductase activity of AIFm2 and facilitates its translocation from mitochondria. His 174 on AIFm2 is the critical target of HNE adduction that triggers this functional switch. HNE adduction and translocation of AIFm2 from mitochondria following I/R injury are attenuated by superoxide dismutase mimetics. These results identify a previously unrecognized role of the MnSOD-HNE-AIFm2 axis, with important consequences for mitochondrial stress signaling, especially in cardiac I/R injury.
Abstract 8: Endothelial TFEB Regulates Postischemic Angiogenesis via AMPKalpha Activation
Authors: Fan Yanbo , Lu Haocheng , Liang Wenying , Guo Yanhong , Zhang Ji , Zhu Tianqing , Hao Yibai , Garcia-Barrio Minerva T , Zhang Jifeng , Chen Y Eugene
Published: July 2021

Rationale:
Postischemic angiogenesis is critical to limit the ischemic tissue damage and improve the blood flow recovery. The regulation and the underlying molecular mechanisms of angiogenesis are not fully unraveled. Transcription factor-EB (TFEB) is emerging as a master gene for autophagy and lysosome biogenesis. However, the role of TFEB in the vascular disease is less understood.

Objective:
We aim to determine the role of endothelial TFEB in postischemic angiogenesis and underlying molecular mechanism.

Methods and Results:
In a murine hindlimb ischemic model, we demonstrated that TFEB was upregulated in the ischemic skeletal muscle tissue. Utilizing genetically-engineered endothelial cell (EC) specific TFEB transgenic mice, we investigated the function of TFEB in postischemic angiogenesis. We observed improved blood perfusion and increased capillary density in the EC-specific TFEB transgenic mice compared with the wild-type littermates (n = 8-9 for each group, p < 0.01). Furthermore, we found that blood flow recovery was attenuated in EC-selective TFEB deficient mice compared with control mice (n =8-9 for each group, p< 0.01). In aortic ring cultures, we found that TFEB transgene significantly increased the vessel sprouting. Adenovirus-mediated TFEB overexpression promoted EC tube formation whereas small interfering RNA (siRNA)-mediated TFEB knockdown suppressed tube formation in ECs. Mechanistically, TFEB activated calcium/calmodulin-dependent protein kinase kinase-β and AMP-activated protein kinase (AMPK)-α signaling pathway. Through pharmacological inactivation and siRNA-mediated knockdown of AMPKα, we demonstrated that AMPKα is necessary for TFEB to regulate tube formation in ECs.

Conclusions:
In summary, our data demonstrate that TFEB is a positive regulator of angiogenesis through activation of AMPKα signaling, suggesting that TFEB constitutes a novel molecular target for ischemic vascular disease.
Abstract 93: Endothelial A Disintegrin and Metalloprotease 10 Deficiency Enhances Murine Atherosclerosis Development
Authors: Theodorou Kosta , van der Vorst Emiel , Rademakers Timo , van Buul Jaap , Lehrke Michael , Lebherz Corinna , Dreymüller Daniela , Ludwig Andreas , Bentzon Jacob , Biessen Erik , Donners Marjo
Published: July 2021

Through shedding of various membrane molecules, including adhesion molecules and chemokines, A Disintegrin And Metalloproteinase 10 (ADAM10) could regulate endothelial permeability and leukocyte recruitment, critical processes in inflammatory diseases like atherosclerosis. Indeed, proteomic analysis on mouse endothelial cell sheddome revealed ±300 differentially regulated proteins upon ADAM10 inhibition, of which 10% appeared involved in permeability and leukocyte transmigration. Accordingly,
in vitro
inhibition of endothelial ADAM10 decreased neutrophil adhesion and transmigration under flow. To evaluate the causal role of endothelial ADAM10 in atherosclerosis development, we used wildtype or endothelial ADAM10-deficient (ADAM10
fl/fl
/Tie2-Cre; in brief ADAM10
del
) mice. Mice were rendered atherogenic by adeno-associated virus-mediated overexpression of PCSK9, resulting in persistent LDL receptor knockdown and hyperlipidemia after high cholesterol diet feeding (HCD). Surprisingly, after 12 weeks of HCD diet feeding, ADAM10
del
mice showed significantly larger (±45%) and more advanced atherosclerotic lesions, with intraplaque hemorrhage in the brachiocephalic artery. Necrotic core area was increased (±87%) and macrophage content decreased (±49%). No differences were observed in granulocyte and collagen content. In contrast to the
in vitro
findings,
in vivo
endothelial permeability, leukocyte adhesion and extravasation, as assessed by intravital multiphoton microscopy, were all increased. In conclusion, this study reveals an unexpected protective effect of endothelial ADAM10 in atherosclerosis development. The underlying mechanisms remain to be determined.
Abstract P099: Soy Food, Bean, Total Legume Consumption and Blood Pressure: The INTERMAP Study
Authors: Oude Griep Linda M , Stamler Jeremiah , Chan Queenie , van Horn Linda , Steffen Lyn M , Miura Katsuyuki , Ueshima Hirotsugu , Okuda Nagako , Zhao Liancheng , Daviglus Martha L , Elliott Paul ,   for the INTERMAP Rsch Group
Published: July 2021

Background:
Legumes, including soy foods and beans, are nutrient-dense high-quality sources of vegetable protein. Evidence from prospective cohort studies suggests beneficial associations between legume consumption and cardiovascular diseases, however, little is known about their impact on blood pressure (BP). Our objective was to quantify associations with BP of consumption of soy food, beans, and total legume consumption, each considered separately.

Methods:
We used cross-sectional data from the International Study of Macro-/Micronutrients and Blood Pressure (INTERMAP) on 4,680 men and women ages 40-59 years from Japan, China, United Kingdom, and United States. During four visits, eight BP and four 24-hr dietary recalls were collected. Regression coefficients per 2SD higher soy food, bean, and total legume intake were estimated using multivariable linear regression models. To estimate overall association, country-specific regression coefficients were pooled, weighted by inverse of their variance. Adjustments were for age, gender, sample, intake of total energy and alcohol, 24-hr urinary sodium excretion, smoking, education, dietary supplement use, adherence to any special diet, history of cardiovascular diseases or diabetes, family history of hypertension, use of antihypertensive, cardiovascular, or diabetes medication, body mass index, and intake of low-fat dairy, vegetables, fibre-rich grain products, red and processed meats, fish and shellfish, and mutually for soy foods or beans.

Results:
Average daily legume intakes (g/1000 kcal) were 69 in Asian participants and 19 in Western participants. Soy foods contributed 91% of total legume intake in Asian participants; beans were the main source of legumes (68%) in Western countries.

In Asian participants, high correlation coefficients were found between intake of soy foods and iron (r=0.34), arginine (r=0.34), aspartic acid (r=0.34), phenylalanine (r=0.32), isoleucine (r=0.30), and magnesium (r=0.30). In Western participants, soy foods were related to dietary magnesium (r=0.30) and beans were highly correlated with dietary fiber (r=0.44) and magnesium (r=0.22).

Total legume intake higher by 76 g/1000 kcal (2SD) was associated with a diastolic BP difference of -0.75 mm Hg (P=0.009) in the total population, -0.70 mm Hg (P=0.05) in Asian, and -0.86 mm Hg (P=0.08) in Western participants. In the total population, diastolic BP differences were -0.62 mm Hg for soy foods (P=0.04) and -0.42 mm Hg for beans and peas (P=0.06). With regard to systolic BP, a significant inverse association was found for soy food intake higher by 41 g/1000 kcal in Western participants (-1.78 mm Hg, P=0.05).

Conclusion:
Higher intake of legumes, especially soy foods, was associated with lower BP levels.
Abstract 015: Critical Periods in Cardiovascular Health Across the Life Course: A Pooled Cohort Analysis
Authors: Allen Norrina B , Krefman Amy , Labarthe Darwin , Greenland Philip , Juonala Markus , Kähönen Mika , Day S. Rena , Lehtimäki Terho , Van Horn Linda , Bazzano Lydia , Liu Lei , Alonso Camilo F , Webber Larry S , Pahkala Katja , Laitinen Tomi , Raitakari Olli , Lloyd-Jones Donald
Published: July 2021

Background:
The prevalence of Ideal Cardiovascular Health (CVH) decreases with age, beginning in childhood. However, more precise estimates of trajectories of CVH across the lifespan are needed to guide intervention. The aims of this analysis are to describe trajectories in CVH from childhood through middle age and examine whether there are critical inflection points in the decline in CVH.

Methods:
We pooled data from five prospective childhood/early adulthood cohorts including Bogalusa, Young Finns, HB!, CARDIA, and STRIP. Clinical CVH factors—blood pressure, BMI, cholesterol, glucose—were categorized as poor, intermediate and ideal then summed to create a clinical CVH score, ranging from 0 to 8 (higher score= more ideal CVH). The association between clinical CVH score and age in years was modeled using a segmented linear mixed model, with a random participant intercept, fixed slopes, and fixed change points. Change points were estimated using an extension of the R package ‘segmented’ which utilizes a likelihood based approach to iteratively determine one or more change points. All models were adjusted for race, gender and cohort.

Results:
This study included 18,290 participants (51% female, 67% White, 46% between the ages of 8-11 at baseline). CVH scores decline with age from 8 through 55 years. We found two ages at which the slope of the CVH trajectories change significantly. CVH scores are generally stable from age 8 until the first change point at age 17 (95% CI 16.3-17.4), when they begin to decline more rapidly with a 0.08 CVH unit loss per year from age 17 to 30. The second change point occurs at age 30 (26.7-33.6) when the rate of decline increases by an additional 0.01 units per year.

Conclusion:
The clinical CVH score declines from favorable levels from childhood through adulthood, with a rapid decline starting at age 17 that becomes slightly steeper from age 30 to 55 years. These inflection points signal that there are critical periods in an individual’s clinical CVH trajectory during which prevention efforts may be targeted.
Abstract MP12: Cardiovascular Health Promoting Behaviors in Older Patients With Chronic Kidney Disease
Authors: Schrauben Sarah , Hsu Jesse Y , Ricardo Ana C , Lash James P , Feldman Harold I , Anderson Amanda H ,   CRIC Study Investigators
Published: July 2021

Background:
The chronic kidney disease (CKD) epidemic continues to grow, and cardiovascular disease (CVD) is the leading cause of death. To modify CVD risk, CKD patients are asked to engage in health promoting behaviors. However, older patients with CKD likely face challenges to engage in healthy behaviors due to social and health factors. This study examined the relationship of social and health factors to health promoting behaviors among younger and older CKD patients and the association of these behaviors with CVD events, death, and CKD progression.

Methods:
Data from the Chronic Renal Insufficiency Cohort (CRIC) Study were analyzed using latent class analysis (LCA) to identify health promoting behavior clusters, stratified by <65 and ≥65 years of age. LCA was based on: BMI of >20 and ≤25kg/m
2
vs. other, healthy diet vs. not, physical activity ≥150min/week vs. not, blood pressure ≤140/90mmHg vs. greater, never/past smoker vs. current, and <7.0% hemoglobin A1c vs. greater. Social factors (self-efficacy, social support, education, income, insurance) and health factors (depressive symptoms, cognition, co-morbidities) were measured by validated surveys and self-report. Logistic regression assessed the association of social and health factors to the behavior clusters. Cox proportional hazards models estimated risk of clusters to CVD events (myocardial infarction/revascularization, peripheral arterial disease, or stroke), CKD progression (incident end-stage renal disease or 50% decline in eGFR), and death from any cause.

Results:
All social and health factors significantly differed between age groups. Three clusters with varying levels of engagement in health promoting behaviors were identified separately among <65 and ≥65 years of age. Among <65 years, the cluster with the highest level of engagement in healthy behaviors was associated with more self-efficacy and lower depressive symptoms. In this age group, in multivariable adjusted models, the clusters with less healthy behavior engagement had a statistically significant increased risk of CVD events (32-81%), death (29-78%), and CKD progression (32-38%). Among ≥65 years, the cluster with the highest level of engagement in healthy behaviors was associated with higher self-efficacy, social support, cognition, and less depressive symptoms. In this age group, in multivariable adjusted models, the clusters with less healthy behaviors had a statistically significant 49% increased risk of death.

Conclusion:
This study demonstrated three clusters of health promoting behaviors that distinguish risk for CVD and other outcomes among older and younger CKD patients. These clusters could identify high-risk groups and be targeted for aggressive management. Clusters with less health promoting behaviors were associated with self-efficacy and depressive symptoms, which could serve as potential targets for intervention.
Abstract MP32: Muscular Strength and Type 2 Diabetes Prevention
Authors: Lee Duck-chul , Brellenthin Angelique , Sui Xuemei , Blair Steven
Published: July 2021

Introduction:
There is little evidence on the effects of muscular strength, independent of cardiorespiratory fitness, on the development of type 2 diabetes.

Hypothesis:
We hypothesised that muscular strength, independent of cardiorespiratory fitness, has significant benefits in type 2 diabetes prevention.

Methods:
Participants were 5,578 men and women aged 18 to 100 years (mean age, 44) who received preventive medical examinations during 1980-2006 in the Aerobics Center Longitudinal Study. Participants were free of myocardial infarction, stroke, cancer, and diabetes at baseline. Total body muscular strength was quantified by combining 1 repetition maximum (1-RM) measures for leg and bench presses and categorized into three groups, lower (weak), middle, and upper (strong), based on the tertiles (thirds) of muscular strength. Cardiorespiratory fitness was measured by a maximal treadmill exercise test. Type 2 diabetes was defined as a fasting plasma glucose level of ≥126 mg/dl, a history of diabetes, or current insulin therapy. Cox regression was used to calculate hazard ratios (HRs) and 95% confidence intervals (CIs) of incident type 2 diabetes by muscular strength after adjusting for baseline age, sex, examination year, body mass index (BMI), current smoking, heavy alcohol drinking, parental history of diabetes, hypertension, hypercholesterolemia, glucose level, and maximum treadmill test time.

Results:
During an average follow-up of 8 years, 270 (4.8%) individuals developed type 2 diabetes. Compared with the individuals in the lower third of muscular strength, individuals in the middle third had a 30% lower risk of developing type 2 diabetes (HR: 0.70, 95% CI: 0.51-0.95), whereas no significant benefit was found in individuals in the upper third (HR: 1.16, 95% CI: 0.86-1.57) after adjusting for potential confounders and cardiorespiratory fitness. We found similar results in men and women, young (<50 years) and old (≥50 years), and normal (BMI <25 kg/m
2
) and overweight/obese (BMI ≥25 kg/m
2
) individuals. In the combined analysis of muscular strength and cardiorespiratory fitness, we also found similar results that individuals in the middle third of muscular strength showed lower risks of developing type 2 diabetes in both low (lower 50%) and high (upper 50%) cardiorespiratory fitness, compared with individuals in the lower third of muscular strength and low cardiorespiratory fitness.

Conclusions:
We found that a moderate level of muscular strength, independent of cardiorespiratory fitness, is associated with a lower risk of developing type 2 diabetes. Additional studies on the dose-response relationship of muscular strength and incident type 2 diabetes are warranted.
Abstract P044: Plasma Phospholipid Very Long Chain Saturated Fatty Acids and Healthy Aging in Older Adults: The Cardiovascular Health Study
Authors: Lemaitre Rozenn N , Biggs Mary L , McKnight Barbara , Lai Heidi T , Song Xiaoling , King Irena B , Psaty Bruce M , Siscovick David S , Mozaffarian Dariush
Published: July 2021

Introduction:
Circulating very long-chain saturated fatty acids (VLSFA) are biomarkers of diet and metabolism which impart different biological activities to sphingolipids and may influence physiological pathways relevant to aging. Higher levels of circulating VLSFAs have been associated with lower risk of all-cause mortality, cardiovascular outcomes, and diabetes. Apart from these associations, it is unknown whether these biomarkers are related to the maintenance of healthy aging.

Hypothesis:
Higher levels of circulating VLSFA are associated with lower risk of unhealthy aging.

Methods:
Phospholipid fatty acids were measured in serial plasma samples collected among participants of the Cardiovascular Health Study (CHS) in 1992-93, 1998-99, and 2005-06. Incident unhealthy aging was defined as the onset of cardiovascular disease (CVD), cancer, lung disease, severe chronic kidney disease, ≥1 difficulties with activities of daily living (ADL), or reduced cognitive function (Mini-Mental State Examination<80). We assessed the associations of arachidic acid (20:0), behenic acid (22:0) and lignoceric acid (24:0), with risk of incident unhealthy aging using Cox regression with time-updated fatty acid levels and covariates.

Results:
Among 2,183 CHS participants with one or more fatty acid measure and healthy aging up to the time of their first VLSFA measurement, mean age at baseline was 75 years, 62% were female, and 12% were black. During median follow-up of 8 years, all but 198 participants failed to age successfully. The most common conditions marking the first occurrence of unhealthy aging were ADL difficulties and CVD (27% each). In analyses adjusted for major risk factors, higher levels of 22:0 and 24:0 were associated with lower risk of unhealthy aging (Table).

Conclusions:
Higher levels of two circulating VLSFA, 22:0 and 24:0, are associated with lower risk of unhealthy aging. These findings open a new avenue of research into these saturated fatty acids and their possible beneficial role in promoting better aging.
Abstract P115: Race, Sex, and Geographical Variation in Perceived Racial Discrimination: The Coronary Artery Risk Development in YoungAdults (CARDIA) Study
Authors: Bey Ganga S , Person Sharina D , Kiefe Catarina I
Published: July 2021

Introduction:
Variation in exposure to discrimination has been proposed as a contributor to disparities in cardiovascular disease (CVD) among black and white women and men in the U.S. Yet, evidence is conflicting, perhaps due to insufficiently studied race-sex and geographical differences in the pathogenicity of discrimination. We hypothesized that the prevalence of perceived racial discrimination in a variety of settings differs by race, sex, and geographic location.

Methods:
We used data from CARDIA, a population-based cohort of black and white adults recruited in Birmingham, AL; Chicago, IL; Minneapolis, MN, and Oakland, CA in 1985-6 (year 0). Racial discrimination perceived in several scenarios was assessed using the Lifetime Discrimination Scale at years 7, 15, and 20 (2005-6). We assessed the prevalence at each of these exams and in each scenario, stratified by race-sex group.

Results:
Prevalence of reported discrimination did not differ markedly over time; we report on year 7 only (n=4,025, figure), with qualitatively similar findings at the other years. Reported discrimination in ≥2 scenarios ranged from 52% in Birmingham to 70% in Minneapolis among black women; among black men, from 65% in Birmingham and 75% in Minneapolis and Oakland. This prevalence was <20% among white women and men in every city. Within all groups, discrimination on the street or in a public setting was most prevalent (p<0.001) and least prevalent in receiving medical care (p<0.001). The proportion of black men reporting discrimination by the police or courts was substantially greater than the other three race-sex groups in each of the four cities (all p<0.001).

Conclusions:
We found variation in the prevalence of reported racial discrimination across race, sex, and geography. Differences in the experience of discrimination may lead to differences in the health-damaging effects of exposure and partially explain inconsistencies in the evidence of discrimination as a cause of disparities in cardiovascular disease between black and white women and men.
Abstract P119: High Stress Reasons for Immigration Affect Diabetes Risk, Body Size and Behavior: The Africans in America Study
Authors: Briker Sara M , Kabakambira J Damascene , Baker Rafeal L , DuBose Christopher W , Mabundo Lilian S , Thomas Francine , Chung Stephanie T , Sumner Anne E
Published: July 2021

Introduction:
Leaving Africa to live in the United States is stressful. Due to oversecretion of glucocortoids and catecholamines, stress has biologic consequences. No data exists on whether Africans who come to America for reasons associated with high stress have a higher rate of conditions such as obesity, diabetes and hypertension than Africans who come for low stress reasons.

Goals:
Working with the Africans in America cohort and using allostatic load score (ALS) to measure biologic stress, our objectives were to (a) rank by ALS the 5 most common reasons for immigration (work, study, asylum/refugee, family unification and lottery); (b) divide the cohort in to 2 groups: high stress (combining the individuals with the 3 reasons with the highest ALS) and low stress (combining the individuals with the lowest ALS); (c) compare body size, visceral adipose tissue (VAT), glucose, BP, sleep duration and smoking in Africans with high and low stress.

Methods:
OGTT and abdominal CT scans were performed in 114 African immigrants (68% male, age 41±10, range22-62y) born in subSaharan Africa but living in Washington, DC.

Results:
Ranked from highest to lowest, ALS for each reason for immigration was: work 3.18±1.71; asylum/refugee 2.68±1.46; study 2.66±1.88; family 1.77±1.27; lottery 1.60±1.27. Hence, high stress reasons for immigration were: work, asylum/refugee and study. Low stress reasons were: family and lottery. ALS in high vs. low stress reasons for immigration were: 2.81±1.74 vs. 1.72±1.25, P<0.01. BP did not vary by group. However, Africans with high stress had a higher BMI, more VAT and hyperglycemia than Africans with low stress (Table). Africans with high stress also had a greater tendency to sleep <7h per night and smoke (Table).

Conclusions:
High stress reasons for immigration include work, asylum/refugee and study. As obesity, hyperglycemia, and behaviors such as decreased sleep duration and smoking are associated with high stress reasons for immigration, medical histories when possible should include reason for immigration.
Abstract P122: Racial/Ethnic Variation in Incidence and Progression of Coronary Artery Calcification: Implications for Understanding the Hispanic Paradox
Authors: Albrecht Sandra S , Lutsey Pamela L , Allison Matthew , Seeman Teresa , Daviglus Martha L , Watson Karol , Polak Joseph F , Rodriguez Carlos J
Published: July 2021

Background:
Previous studies show that Hispanic persons have similar or lower levels of coronary artery calcium (CAC) and slower progression than non-Hispanic whites (NHW), even after adjustment for traditional risk factors. We examined whether this health advantage in CAC incidence and progression among Hispanic adults extends across all levels of risk factor (RF) burden, and whether associations vary by nativity (foreign-born, US-born) and by heritage group (Mexican, non-Mexican).

Methods:
We analyzed data on Hispanic and NHW participants aged 45-84 years from the Multi-Ethnic Study of Atherosclerosis (MESA). Follow-up CAC measurements and complete covariate data were available for 3694 participants with an average of 6.6 years between the follow-up and baseline scans (2000-2002). Baseline measures of the following traditional RFs were considered: current cigarette smoking, high total cholesterol, hypertension, diabetes, and obesity, with RF burden scores ranging from 0-5. Outcomes were incident CAC (any follow-up CAC >0 Agatston units) among individuals without detectable CAC at baseline, and CAC progression (any positive increase in CAC) among all participants estimated using relative risk regression. All models were adjusted for age, sex, RF burden, race/ethnicity, education, income, and time between scans

Results:
Although a higher proportion of Hispanics had RF burden scores ≥3 compared to NHW (14.6% vs 8.9%, p<0.0001), Hispanics had a lower adjusted incidence (risk ratio (RR) = 0.83, 95% CI: 0.72-0.96) and less progression of CAC (RR=0.90, 95% CI: 0.86-0.95) than NHW. However, there was evidence of heterogeneity in this pattern. For example, among individuals with no detectable baseline CAC, a Hispanic health advantage was only seen among individuals with RF burden scores of 0 (RR=0.66, 95% CI: 0.48-0.91 for Hispanics vs. NHW at RF=0), with race/ethnic differences getting progressively smaller with increasing RF burden (for RF ≥3: RR=1.01, 95% CI: 0.69-1.48). Compared to NHW, lower adjusted incidence and progression of CAC was evident to an even greater extent among foreign-born Hispanics, but a health advantage was still present for US-born Hispanics, and for both Hispanic heritage groups. However, these patterns also only remained among individuals with lower RF burden scores.

Conclusions:
The Hispanic health advantage in CAC incidence and progression was primarily evident among individuals with fewer traditional risk factors for CVD, but was present among different Hispanics groups. Future research is necessary to identify the factors underlying this advantage, and the dynamics that erode it as RF burden increases.
Abstract P125: Multiple Measures of Energy Expenditure and Substrate Oxidation in Non-obese White and African American Young Adults
Authors: Katzmarzyk Peter T , Ravussin Eric
Published: July 2021

Introduction:
African Americans (AA) experience higher rates of obesity and related disorders than the general U.S. population. It has been hypothesized that the increased risk of obesity among AA may be explained, in part, by lower levels of energy expenditure (EE) and lower levels of fat oxidation. However, many different measures of EE and substrate oxidation have been employed across previous studies.

Objective:
The objective of this study was to compare multiple measures of EE and substrate oxidation among White (W) and AA adults. We hypothesize that AA will have lower EE and lower fat oxidation rates than W.

Methods:
A sample of 12 young (ages 22 to 35 y), non-obese AA adults was recruited from the local community and pair-matched by age, sex and body mass index (BMI) to a sample of 12 W adults. Height and weight were measured and BMI was calculated (kg/m
2
). Total fat mass (FM) and fat free mass (FFM) were measured using dual energy x-ray absorptiometry. Resting EE (REE) and respiratory quotient (RQ) were measured in a fasting state using a metabolic cart; 24-hour EE, 24-h RQ, sleep EE and sleep RQ were measured in a whole room calorimeter; and free-living total daily EE (TDEE) was measured over two weeks using doubly labelled water. Physical activity level (PAL) was computed as TDEE/REE. Differences between W and AA were determined using general linear models, adjusting for FFM.

Results:
The analytic sample had a mean age of 27.0 y (SD 4.3 y) and mean BMI of 22.9 kg/m
2
(SD 2.9 kg/m
2
). There were no significant differences in age, BMI, FM or FFM between W and AA (all p>0.05). However, W had significantly higher REE (1459 vs 1305 kcal/day; p=0.001), 24-h EE (1826 versus 1737 kcal/day; p=0.02), sleep EE (1509 vs 1405 kcal/day; p=0.005); but not TDEE (2452 vs 2313 kcal/day; p=0.30) compared to AA. There were no race differences in RQ (0.83 vs 0.83; p=0.93), 24-h RQ (0.86 vs 0.88; p=0.24) or sleep RQ (0.86 vs 0.87; p=0.44). On the other hand, AA had higher PAL (1.34 vs 1.26; p=0.04) compared to W.

Conclusions:
Non-obese W adults demonstrated higher REE, 24-h EE, and sleep EE compared to AA, but had similar levels of free-living TDEE. It appears as though some AA adults may compensate for lower REE by increased physical activity, which may be an effective strategy to prevent weight gain and obesity.
Abstract P176: Moderate Physical Activity and Sleep Duration as Predictors of Cognitive Function in Older Adults With Hypertension: NHANES 2011-2014
Authors: Hoffmann Nicole , Petrov Megan E
Published: July 2021

Introduction:
Hypertension is associated with increased risk for cognitive decline. Lifestyle behaviors such as moderate physical activity (MPA) and adequate sleep duration may mitigate this decline, though limited research exists. The aim of the study was to examine the joint association of MPA and sleep duration on cognitive function by hypertension status.

Methods:
Adults (n=2976, ≥60yrs) from the 2011-2014 National Health and Nutrition Examination Survey (NHANES) were assessed for their habitual sleep duration (<7, 7-8.9,
≥9hr)
, self-reported participation in regular MPA (yes/no), reported physician-diagnosed hypertension (yes/no), and cognitive function (Digit Symbol Substitution Test [DSST]; Animal Fluency test). Weighted linear regression analyses were conducted to assess joint association of sleep duration and MPA on cognitive function, and test the modifying effect of hypertension status (alpha level set at 0.1) after adjustment for demographics.

Results:
See Table. There were significant main effects for combined MPA and sleep duration on DSST (Wald
F
(5,28)=5.33,
p
=.001) and Animal Fluency (Wald
F
(5,28)=2.58,
p
=.05). Participants who did not engage in MPA regardless of sleep duration had significantly worse cognitive function compared to participants who engaged in MPA and obtained 7-8.9hr sleep. There was a significant interaction between MPA-sleep duration groups and hypertension status on DSST (Wald
F
(5,28)=2.42,
p
=.06), but not on Animal Fluency. Stratified analyses indicated among individuals with hypertension the buffering effect of MPA regardless of sleep duration was maintained, but not for individuals without hypertension.

Conclusions:
In a sample of adults, regular MPA predicted better cognitive outcomes regardless of sleep duration. Among individuals with hypertension regular MPA regardless of sleep duration was significantly associated with better executive function, but no such association was found among individuals without hypertension.
Abstract P234: Dairy Fat Intake and Risk of Type 2 Diabetes in 3 Cohorts of US Adults
Authors: Ardisson Korat Andres , Hu Frank B , Sun Qi
Published: July 2021

Background:
Previous studies have examined the association between dairy fat intake and incident Type 2 Diabetes (T2D) by conducting analyses of dairy products stratified by fat content, although data linking dairy fat intake and incident T2D and their substitution for other nutrients are sparse.

Objective:
The aim of this study is to evaluate the association between dairy fat intake and risk of T2D. We assessed the hypothesis that replacing calories from dairy fat for other animal fat or refined carbohydrates will result in modest increases in T2D risk.

Methods:
We followed up 41,670 men in the Health Professionals Follow-Up Study (1986-2010), 84, 685 women in the Nurses’ Health Study (NHS; 1980-2012), and 90,325 women in the NHSII (1991-2011). Diet was assessed every 4 years with the use of validated food-frequency questionnaires, and other health and lifestyle covariates were collected biennially. Dairy fat contents were determined for dairy products and food items that contain dairy. Dairy fat intake from all relevant food items was summed to calculate total intake, which was expressed as percent of total energy. Incident T2D cases were identified by self-reports during follow-up and confirmed by a validated supplementary questionnaire. A time-dependent Cox proportional hazards regression was used to estimate the hazard ratio for dairy fat intake and T2D risk.

Results:
During 4,661,518 years of follow-up, we documented 18,298 incident T2D cases. In multivariate models, a 5% increase in energy dairy fat was associated with a 2% risk increase in T2D (RR: 1.02; 95% CI: 1.00, 1.05). In isocaloric substitution models, the replacement of 5% of calories from dairy fat with the equivalent energy from other sources of animal fat or carbohydrate from refined grains was associated with an 7% [RR: 1.07; 95% CI: 1.04, 1.09], and a 7% [RR: 1.07; 95% CI: 1.04, 1.11] increased risk of T2D, respectively. Conversely, a 5% calorie substitution of carbohydrate from whole grains was associated with 7% lower risk of T2D [RR: 0.93; 95% CI: 0.89, 0.97].

Conclusions:
In conclusion, dairy fat intake was modestly associated with a higher T2D risk. The replacement of dairy fat with carbohydrates from whole grains may decrease incident T2D risk. Further research is warranted to elucidate the role of other components in dairy products that may contribute to previously reported null associations with T2D.
Abstract P274: Cross-sectional Analysis of Baseline Characteristics for New Users of Vorapaxar (PAR-1 Antagonist) and P2Y
12
Inhibitors
Authors: Pinto Cathy Anne , Hess Gregory P. , Fonseca Eileen , Acquavella John
Published: July 2021

Introduction:
Vorapaxar is indicated for the reduction of thrombotic events in patients with a prior myocardial infarction (MI) or peripheral arterial disease (PAD), and contraindicated in patients with a prior stroke, transient ischemic attack (TIA), or intracranial hemorrhage (ICH). This is the first descriptive study of vorapaxar in US clinical practice.

Objectives:
Main objective was to describe baseline characteristics of vorapaxar new users. For context, P2Y
12
new users were also examined, with indications summarized in Table 1.

Methods:
Cross-sectional analysis of vorapaxar new users ≥18 years with ≥1 prescription between June 2014 - June 2016 identified in Symphony Health’s outpatient pharmacy claims database linked to private practitioner claims and hospital data. Concomitant clopidogrel use was defined with overlapping days of supply. Medical history was defined using a 1-year baseline period.

Results:
The analysis included 1,362 vorapaxar (44% with clopidogrel), 975,023 clopidogrel, 133,724 ticagrelor, and 98,101 prasugrel new users (Table 1). Most ticagrelor (87%) and prasugrel (85%) users had evidence of coronary artery disease (CAD) compared with 52% of clopidogrel and 58% of vorapaxar users. In contrast, <1% of ticagrelor and prasugrel users had PAD but no CAD vs. 6% of clopidogrel and 15% of vorapaxar users. The proportion of vorapaxar users (44%) with PAD, alone or with CAD, was 3-5 times larger than P2Y
12
users. PAD was more prevalent for vorapaxar with (50%) vs. without (39%) clopidogrel. Contraindications were infrequent for vorapaxar users - no ICH, <4% with stroke or TIA. Median time from recent MI to therapy start was 6 months for vorapaxar vs. 3 days for ticagrelor and prasugrel and 7 days for clopidogrel. Vorapaxar users were mostly male and on average ~65 years old.

Conclusions:
The majority of vorapaxar new users had a history of CAD and/or PAD, with PAD prevalence more than twice that of P2Y
12
new users. Contraindications were infrequent.