bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 15203: Single Cell Transcriptional Analysis Implicates
Acta2
Positive Pericytes in Arterial Muscularization and Imbalance in Bmp and Angpt Signaling During Development of Pulmonary Arterial Hypertension
Authors: Cober Nicholas D , McCourt Emma , Soares Godoy Rafael , Deng Yupu , Schlosser Ken , Cook David , Wang Liyuan , Stewart Duncan J
Published: June 2023

Introduction:
Pulmonary arterial hypertension (PAH) is a lethal disease characterized by marked vascular remodeling, inflammation and increased pulmonary vascular resistance. Pericytes normally play a key role in maintaining microvascular homeostasis; however, they can also contribute to increased muscularization of distal arterioles in PAH by giving rise to smooth cells (SMCs).

Methods:
Sprague-Dawley rats were injected with 20mg/kg SU subcutaneously and exposed to 3 weeks of chronic hypoxia (CH, 10% O2). Right ventricular systolic pressure (RVSP) was measured at baseline, 1, 3, 5, and 8-weeks, then lungs were explanted, digested, and dissociated into single cells which were sequenced using the 10x genomics platform.

Results:
RVSP was significantly elevated at 1 week after SU (47mmHg) reaching a plateau of >100 mmHg by 5 weeks. Dimensionality reduction of the scRNAseq data was performed using Uniform Manifold Approximation and Projection (UMAP) analysis resulting in 24 lung clusters and sub-clustering of stomal populations yielded seven clusters. In addition to fibroblasts, SMC/myofibroblasts and mesothelial cells, we identified 2 distinct pericyte clusters both expressing canonical pericyte markers such as
Pdgfrb
and
Csgp4
(
Ng2
) and differentiated by presence or absence of
Acta2
.
Acta2
+
pericytes exhibited high expression of contractile markers (i.e.,
Myh11
and
Tagln
), consistent with partial SMC differentiation, and underwent a substantial (>3-fold) expansion during PAH development. The

Acta2
+

pericytes also exhibited profound downregulation in the endothelial protective gene,
Bmpr2
, with a reciprocal upregulation of
Rbpms2
, a gene which inhibits BMP signaling. Similarly, reciprocal reduction in
Angpt1
, an agonist of the endothelial Tie2 receptor and an increase in expression the partial Tie2 antagonist,
Angpt2
, thereby favoring EC activation and inflammation.

Conclusion:
These results suggest expansion of
Acta2
+
pericytes in PAH may contribute to distal arteriolar muscularization as well as an imbalance in BMP/TGFß and ANGPT1/2 signaling, promoting EC apoptosis, activation, and inflammation in the SU/CH model.
Abstract 15238: Impact of Statins on All-Cause Mortality in Subjects With Severe Coronary Artery Calcium Score: Results Form the Confirm (Coronary Ct Angiography Evaluation for Clinical Outcomes: An International Multicenter Registry) Registry
Authors: Verghese Dhiran , Kinninger April , Manubolu Venkat S , Aldana-Bitar Jairo , Alalawi Luay Hussein , Min James K , Earls James , Achenbach Stephan , Dunham Ashley , Crabtree Tami , Gransar Heidi , Al-Mallah Mouaz , Bax Jeroen J , Berman Daniel S , Cademartiri Filippo , Callister Tracy , Chang Hyuk-Jae , Chow Benjamin J , Cury Ricardo C , Feuchtner Gudrun M , Hadamitzky Martin , Hausleiter Joerg , Kaufmann Philipp A , Leipsic Jonathon , Lin Fay Y , Kim Yong-Jin , Marques Hugo , Pontone Gianluca , Rubinshtein Ronen , Villines Todd , Shaw Leslee J , Budoff Matthew ,   CONFIRM Investigators
Published: June 2023

Introduction:
Coronary artery calcium (CAC) score improves prognostic accuracy of atherosclerotic cardiovascular disease outcomes. However, the relative impact of statins on outcomes stratified by CAC score is limited.

Methods:
The open-label, 12-center, 6-country prospective longitudinal CONFIRM registry was queried. Individuals with severe CAC (≥300), with long term follow up were selected. Subjects with prior ASCVD events were excluded. The primary study end point was all-cause mortality (ACM) which was compared in individuals on stain versus those not on statin. Kaplan-Meier cumulative incidence curves were performed and compared with log rank test. Multivariate Cox proportional hazard regression analysis was used to calculate hazard ratio (HR) with 95% confidence interval (CI).

Results:
Of 21,863 participants from the CONFRIM registry, 5,928 had no prior ASCVD events and had information on CAC and statin use. Of these, 972 had a CAC score of ≥300. The mean age of statin users and non-users with CAC≥300 was similar at 67.7± 9.0 vs. 66.9±10.5; p=0.166, and both groups were predominantly male (70% vs 69%, p=0.736). Statin users were more likely to have high cholesterol (93% vs 43%), hypertension (74% vs. 67%), diabetes mellitus (27% vs 20%), family history of cardiovascular disease (41% vs. 34%) and less likely to be a current smoker (22% vs 34%); all p<0.05. There were 42 (9%) deaths over a mean follow-up of 4.2 years among statin users and 86 (18%) deaths over 3.9 years in non-users. In multivariable proportional hazard Cox modeling with a reference group of non-statin users, statin participants with CAC of ≥300 had significantly reduced risk for ACM, HR 0.52- (95% CI, 0.33-0.82, p=0.005).

Conclusions:
In this large prospective registry, in the primary prevention cohort of patients without prior ASCVD events, severe CAC ≥ 300 identified patients most likely to benefit from statin use. Statins lowered ACM in patients with CAC≥ 300 by 48%.
Abstract 15242: Donor-Derived Chagas Disease Masquerading as Acute Heart Transplant Rejection
Authors: Jafree Ehsan , Botheju W. Savindu Pasan , Mao Yuxuan , Tomdio Anna , Gentry Kristyn
Published: June 2023

Introduction:
Although Chagas disease affects over 300,000 residents in the US, donor-derived Trypanosoma cruzi transmission is an uncommon complication after heart transplantation. We present a case of post-transplant Chagas cardiomyopathy.

Results:
A 68 year old male with history of heart failure with reduced ejection fraction secondary to ischemic cardiomyopathy, status post HeartMate III and subsequent orthotopic heart transplantation from a Hispanic donor, initially presented four months after transplant for routine right heart catheterization. Endomyocardial biopsy showed grade 2R acute cellular rejection. Patient was treated with IV steroids and discharged. One month later, he presented with one week of fevers, malaise, non-tender erythematous patches on his upper body and hemorrhagic bullae on his buccal mucosa. Initial workup was notable for pancytopenia with WBC 1.3 x10e9/L, hemoglobin 10.3g/dL, and platelets 77 x10e9/L, troponin 3.47 ng/mL, NT-proBNP 11,331 pg/mL, and new right bundle branch block on electrocardiogram. Cardiac MRI showed extensive multifocal late gadolinium enhancement and pericardial exudates, suggestive of atypical infection. Despite treatment with Filgrastim and IVIG, the patient progressed to cardiogenic shock complicated by complete heart block requiring transvenous pacing, and ventricular tachycardia. On hospital day 5, a peripheral smear confirmed Trypanosoma cruzi with high parasite burden and amastigotes were visualized on a shave biopsy of his rash. Patient was initiated on Benznidazole. He was treated in the intensive care unit, where his course was further complicated by recurrent VT, for which he received an implantable cardiac defibrillator, and renal failure requiring hemodialysis.

Conclusions:
We present a case of suspected donor-derived Chagas Disease. In retrospect, this patient’s initial episode of acute rejection was most likely due to Chagas Disease. Due to increasing globalization of endemic diseases and life-threatening complications associated with this disease, a high index of suspicion is required to diagnose and treat T. cruzi after heart transplantation in the setting of non-specific symptoms.
Abstract 15253: Mitochondrial Trifunctional Protein HADHA Regulates Mitochondrial Cardiolipin Content in Right Ventricle in Pulmonary Hypertension
Authors: Zimmer Alexsandra , Zhang Peng , Wang Eric , Yalcin Emine , D'Silva Natalie , Clements Richard T , Choudhary Gaurav
Published: June 2023

Background:
Pulmonary hypertension (PH) is prevalent in cardiopulmonary and metabolic diseases. Unmet energy needs due to mitochondrial (mito) dysfunction results in myocardial contractile failure, as seen in the dysfunctional right ventricles (RV). Cardiolipin (CL) is a critical lipid for mito structure and cristae organization that are essential for mito oxidative phosphorylation and ATP generation. The synthesis and maturation of CL involves many steps including a final step where acylation of monolysocardiolipin occurs and CL[18:1]
2
[18:2]
2
is converted to mature CL[18:2]
4
catalyzed by a mito trifunctional protein, HADHA. To date, the expression of mature CL and its regulation by HADHA in RV in PH remains unknown.

Hypothesis:
We hypothesize that HADHA regulates mito CL contents in RV in PH.

Methods:
Adult Sprague Dawley rats received 20 mg/kg sc SU5416 were exposed to 3 wks of hypoxia (10% FiO2), followed by 4 wks of normoxia. Control rats injected with vehicle were kept in normoxia. Hemodynamic and echocardiography were performed, followed by HADHA and CL analysis. We also generated
in vitro
models with H9c2 cells exposure in hypoxia and stable HADHA-knockdown H9c2 cells for mechanistic studies.

Results:
PH rats show RV dysfunction. Mito proteomics show a 70% decrease in HADHA expression in mito from RV of PH rats in comparison to the control rats, which is further validated by Western Blot and qPCR. Using mass spectrometry, mature CL (CL[18:2]
4
) in RV of PH rats was found to decrease to 55% of that in RV of control rats (n=4-5/group,
p
<0.05). Exposing the H9c2 cells to hypoxia (1% O
2
) for 72 hrs recapitulate the downregulation of HADHA observed in the dysfunctional RV of PH rats, which is associated with a 70% reduction in the account of mature CL (n=6 each,
p
<0.05). Importantly, adenoviral overexpression of HADHA almost brings the mature CL content in hypoxia to be comparable to cells in normoxia (n=6 each,
p
<0.05). Our data also show that HADHA-specific knockdown is sufficient to decrease mature CL in H9c2 cells (Control 100% vs. HADHA Knockdown 33%, n=3 each,
p
<0.05).

Conclusions:
Our data demonstrate that HADHA is critical in maintaining mature mito CL contents. Downregulation of HADHA contributes to reduced mito CL contents in RV in settings of PH.
Abstract 15263: Trends and Clustering in Cardiovascular Health Metrics by Race and Ethnicity Among US Adults, 2011-2020
Authors: Lu Yuan , Li Xumin , Liu Yuntian , Spatz Erica S , Herrin Jeph , Krumholz Harlan M
Published: June 2023

Introduction:
Nationally representative data evaluating recent trends and clustering in cardiovascular health metrics by race and ethnicity are sparse.

Methods:
We evaluated 10-year trends and clustering in cardiovascular health metrics in 10,300 individuals who attended the morning fasting examination, using data from the National Health and Nutrition Examination Surveys (NHANES) from 2011 to 2020. We assessed the time trends of the mean score of the 7 cardiovascular health metrics by race and ethnicity. Then we ranked the top 10 mutually exclusive phenotypes of the 7 cardiovascular health metrics for each race and ethnicity.

Results:
Among participants, the mean age was 48.3 (SD, 4.2) years; 52% were female; 6.3%, 71.3%, 12.7%, and 9.7% were Asian, White, Black, and Hispanic. From 2011 to 2020, the total cardiovascular health score decreased significantly for Asian individuals (P=0.005) but did not change significantly for Black, Hispanic, and White individuals (P>0.05 for all). The mean scores for individual cardiovascular health metrics did not change by racial/ethnic subgroups, except for a decrease in mean body mass index (BMI) score in Asian individuals, an increase in mean total cholesterol score in Black individuals, a decrease in mean fasting glucose score in Asian, Hispanic, and White individuals. Among all the individuals, healthy diet and the combination of healthy diet and physical activity were the two most prevalent phenotypes, followed by several different combinations of physical activity, BMI, healthy diet, and smoking.

Conclusions:
Temporal trends suggest an overall unchanged mean total cardiovascular health score for Black, Hispanic, and White individuals and a decreased score for Asian individuals. There were persistent racial/ethnic disparities in the individual cardiovascular health metrics and phenotypes. Across all racial/ethnic subgroups, large opportunities exist for improvements.
Abstract 15287: Optimizing Guideline Directed Medical Therapy in Recently Hospitalized Patients With Heart Failure in a Large Urban Veterans Affairs Medical Center
Authors: Moolchandani Priyanka , PATEL SATYA , Saito Ashley , GREGORIO STEPHANIE , Li Jiwen
Published: June 2023

Introduction:
A quarter of patients hospitalized for heart failure either die or are re-hospitalized within 30 days. Optimization of guideline directed medical therapy (GDMT) has been shown to decrease both mortality and hospital readmissions.

Methods:
We used an existing heart failure clinical dashboard registry at our Veterans Affairs (VA) hospital to determine the rates of hospitalization from any medical cause and GDMT prescription rates from November to December 2022. We performed a retrospective chart review to validate dashboard results.

Results:
Twenty-nine patients in the registry were hospitalized within the last 30 days, with 5 patients excluded due to ongoing hospitalization; 19 were classified as heart failure with reduced ejection fraction (HFrEF, LVEF ≤ 40%), 4 as heart failure with preserved ejection fraction (LVEF ≥ 50%), and 2 unknown (no EF in 3 years). The rates of quadruple GDMT was 16% in all heart failure patients and 21% in patients with HFrEF (table 1). We performed a root cause analysis of low GDMT rates (figure 1). We used this data to secure buy-in from hospitalist medicine and cardiology to support a full-time cardiology pharmacist to screen all currently admitted patients with HFrEF. We created a standardized note template that gives recommendations for GDMT optimization prior to discharge.

Conclusions:
Although there is strong evidence that GDMT improves patient outcomes, many recently hospitalized patients are not optimized even after accounting for relative contraindications. We demonstrate that using tools such as a clinical dashboard and problem visualization with quality improvement methodology can both make GDMT optimization more tangible and also create buy-in from the health system to implement change.
Abstract 15298: Paradoxical Association Between Sleep Duration and Risk of Stroke: Analysis of the National Health and Nutrition Examination Survey 2017 - 2020
Authors: Lalitnithi Pakin , Lopimpisuth Chawin , Thimphitthaya Chanattha , Pangkanon Watsachon , Wattanachayakul Phuuwadith , Wannaphut Chalothorn , Tantisattamo Ekamol
Published: June 2023

Introduction:
Although inadequate sleep is known to associate with cardiovascular (CV) outcomes including stroke, association between sleep duration and stroke is unclear.

Hypothesis:
Shorter sleep duration is associated with higher risk of stroke.

Methods:
A retrospective cross-sectional analysis of the 2017-2020 National Health and Nutrition Examination Survey (NHANES) included adult participants aged 21 to 79 years with self-reported history of stroke and sleep duration was studied. Association of sleep duration, stratified into short (<6 hours) and normal sleep durations (≥6 hours), and stroke was evaluated by multivariate logistic regression.

Results:
A total of 8,416 participants were identified, of which mean age was 48.25±16 years old, 51.5% were female and mean sleep duration was 7.529 ± 1.651 hours. Overall, 377 patients (4.5%) had a stroke, of which mean sleep duration was 7.545 ± 2.132. Mean sleep duration of non-stroke population was 7.528 ± 1.626. The difference of mean sleep duration between these two groups was statistically significant (0.017 hours; P < 0.001; 95% Cl -0.156,0.189; Figure1). The majority of the study participants reported a normal sleep duration (7426 participants, 88.2%) and the remaining 913 participants (10.8%) had a short sleep duration. Compared with the normal sleep duration group, participants with the short sleep duration had significantly lesser the odds of stroke (adjusted odds ratio 0.574; P < 0.001; 95% CI 0.424,0.777) after adjusted for baseline risk factors including age, gender, ethnicity, body mass index (<25 vs. ≥25 kg/m
2
), history of hypertension and diabetes.

Conclusions:
Contrary to previous reports, this study shows that sleep duration is positively associated with risk of stroke. Further long-term studies that focus on understanding the pathophysiology using non-self-reported method are required to elucidate effect of sleep quality and optimal sleep duration on the adverse CV outcomes.
Abstract 15332: Post-Resuscitation Epinephrine Infusion Restores Blood Pressure but Not Blood Flow in a Rat Model of Severe Pediatric Cardiac Arrest
Authors: Ton Hoai , velado Victor , Shoykhet Michael
Published: June 2023

Introduction:
In children resuscitated from cardiac arrest (CA), post-arrest hypotension occurs often and worsens outcomes. Continuous infusion of epinephrine (Epi) is the 1st-line treatment for post-arrest hypotension but may worsen organ perfusion due to vasoconstriction.

Hypothesis:
Epi infusion after resuscitation from pediatric asphyxial CA restores blood pressure but not organ perfusion.

Methods:
Immature rats (PND18-22, n=34) underwent 11.5 minutes of asphyxial CA followed by resuscitation. Rats with hypotension (mean arterial pressure (MAP)<80% baseline) after resuscitation received infusions of either 0.9% NaCl (NS) or Epinephrine (Epi). MAP, carotid and femoral arterial blood flows (Fc, Ff) and renal oxygen tension (PrO2) were recorded.

Results:
One hour after resuscitation, MAP, Fc and Ff were 61.73 ± 2.68; 42.58 ± 3.84; and 19.53 ± 7.00 % of baseline, respectively. Epi infusion increased MAP in a dose-dependent manner, reaching 95.13 ± 7.93 % of baseline at 0.5 μg/kg/min. In contrast, Epi infusion up to 0.5 μg/kg/min increased Fc only to 49.55 ± 6.49 and Ff to 53.28 ± 7.13 % of baseline. NS infusion at equivalent volume had a more salutary effect than Epi on improving blood flow (Fc 64.62 ± 6.64 and Ff 60.25 ± 7.60 % of baseline). PrO2 reached 100% of baseline with Epi infusion and inspired O2 fraction (FiO2) = 1. However, at FiO2 = 0.21, Epi failed to restore PrO2 which reached 42% of baseline. Interestingly, a single 10 cc/kg NS bolus increased PrO2 to 100% of baseline at all FiO2 tested.

Conclusions:
In this rat model of severe pediatric asphyxial CA and resuscitation, post-resuscitation Epi infusion failed to restore carotid and femoral arterial blood flow despite normalizing blood pressure. Furthermore, Epi infusion restored neither renal perfusion nor oxygenation. In contrast, infusion of NS at comparable volumes was more effective than Epi at normalizing carotid and femoral blood flows as well as renal oxygenation. If validated in a large animal model, these results may indicate that Epi infusion after CA resuscitation contributes to organ injury and morbidity in children.
Abstract 15374: Somatic Mutations of Dnmt3a Contribute to Pulmonary Vascular Remodeling and Right Ventricular Dysfunction in PAH
Authors: Al-Qazazi Ruaa , Martin Ashley Y , potus francois , Rauh Michael , Archer Stephen L
Published: June 2023

Background:
Pulmonary Arterial Hypertension (PAH) is an obliterative pulmonary vasculopathy in which structural changes are driven by inflammation, fibrosis, mitochondrial dysfunction, and pathological angiogenesis. While ~10% of cases of PAH have germline BMPR2 mutations (or one of 16 other recognized or putative PAH genes), the genetic basis for the majority of PAH patients remains unexplained. Somatic mutations in hematopoietic stem cells (HSC) are related to the disorders of clonal hematopoiesis of indeterminate potential (CHIP). Tet methylcytosine dioxygenase 2 (TET2) and DNA methyltransferase 3a (DNMT3A) are among the primary genes involved in CHIP. We found in a PAH-Biobank that 0.39% of PAH patients had somatic or germline TET2 mutations. Also, TET2-knockout mice can develop PAH spontaneously. Thus TET2 is a new PAH gene. We now hypothesize that CHIP driven by mutations of DNMT3A can also cause pulmonary vascular remodeling and right ventricular failure (RVF), contributing to PAH.

Methods:
Studies were performed on 9 month old DNMT3A knockout (-/-) mice and their relative controls (f/f) (N=5/group). The right ventricular (RV) function was assessed via cardiac ultrasound. Pulmonary artery medial wall thickness (PAMWT) and radial alveolar count (RAC) were assessed in lung tissue sections using light microscopy.

Results:
In DNMT3A -/- mice, there was a reduction in pulmonary artery acceleration time (PAAT) compared to controls (
p=
0.0268). There was also a significant reduction in percent RV free wall thickening (RVFWT %) compared to controls (57.44±9.97 DNMT3A-/-, 118.5±14.98 f/f;
p=
0.017). Both Tricuspid annular plane systolic excursion (TAPSE) and cardiac output (CO) were also significantly reduced in DNMT3A -/- mice (
p
<
0.0001,
p
=
0.0135). The medial thickness of small pulmonary arteries was significantly higher in DNMT3A -/- mice compared to controls (56.51% ±1.19 DNMT3A-/-, 39.79% ±2.07 f/f;
p=
0.002). RAC was significantly reduced in the DNMT3A -/- mice when compared to controls (
p=
0.018).

Conclusions:
DNMT3A mutations may contribute to the development of pulmonary vascular disease and RVF in PAH.
Abstract 15380: Cardiac Localized Polycystin 2 is Required for Natriuretic Peptide Production and Its Absence is a Main Driver of Hypertension
Authors: Lantonio Brandon , Kuo Ivana Y
Published: June 2023

Introduction:
Autosomal Dominant Polycystic Kidney Disease (ADPKD) occurs in 1:1000 births and causes renal cysts and loss of renal function. However, cardiovascular complications are the most common cause of mortality in ADPKD patients and hypertension affects 70% of patients before decline in renal function. ADPKD arises from mutations in
PKD1
and
PKD2,
encoding Polycystin 1 and Polycystin 2 (PC2) respectively. PC2 forms a nonselective cation pore channel that conducts calcium. Heterozygous mice for
Pkd2
have attenuated natriuretic peptide (NP) responses. The NPs, ANP and BNP, are regulators of euvolemia via renal excretion of sodium following cardiac atrial and ventricular stretch, thought to be a calcium activated process. We hypothesized cardiac PC2 contributes to NP production and loss of PC2 induces hypertension without renal involvement.

Methods:
Cardiomyocyte inducible PC2 KO (PC2 cKO) mice were generated by crossing
Pkd2
floxed mice with αMHC-MerCreMer mice. Cardiomyocyte Cre expression was induced by tamoxifen diet at 6 weeks. Mice were aged to 7 months and had intra-aortic blood pressure monitoring or left ventricular tissue collection for quantitative PCR (qPCR) analysis. Male and female mice (n=4-9) were used and analyzed separately by Student’s t-Test, with a p <0.05 considered statistically significant.

Results:
PC2 cKO mice had elevated diurnal blood pressure (160/110mmHg) without renal cysts. Comparisons were made to age-matched controls. All reported genes were affected in males and females, with only female data reported (Mean ± SEM). qPCR analysis showed that PC2 cKO mice had a ~50% decrease in
Nppa
(WT: 1± 0.2 KO: 0.6 ± 0.1) and
Nppb
(WT: 1 ± 0.1 KO: 0.5 ± 0.2) mRNA, genes encoding ANP and BNP respectively. There was a ~60% reduction in
Pcsk6
(WT: 1 ± 0.1, KO: 0.4 ± 0.1) mRNA in PC2 cKO mice, a gene encoding a protease necessary for Corin activity, regulating ANP.
Cgb
mRNA was abolished (WT: 1 ± 0.2, KO: 0.03 ± 0.01) in PC2 cKO mice, encoding Chromogranin B, a regulator of NF-κB mediated production of
Nppb
mRNA and ANP-containing vesicles.

Conclusion:
We conclude loss of PC2 in cardiomyocytes leads to a hypertensive phenotype which may arise from a calcium-dependent multi-component dysfunction of cardiomyocyte NP production.
Abstract 15388: A Randomized Placebo-Controlled Trial of Omega-3 Fatty Acids, Lycopene and Low Sodium Diet
Authors: Moser Debra K , Chung Misook L , Biddle Martha J , Rayens Mary Kay , Lennie Terry
Published: June 2023

Background:
Patients with heart failure (HF) commonly remain symptomatic after medical treatment. Symptoms are associated with rehospitalizations and mortality. We developed a 6-month nutrition intervention targeting the 3 most common HF symptoms: edema, shortness of air, and fatigue. The intervention involves 3 nutrients that target the pathologic pathways underlying symptoms: sodium, omega-3 fatty acids, and lycopene.

Hypothesis:
Time to first event will be longer in the nutrition intervention group than in the placebo group at 1-year follow-up.

Methods:
This was a randomized controlled double-blind clinical trial where 118 patients (mean age 63±12 years; 40% female; 64% NYHA class III or IV) with HF were randomized to active intervention vs placebo groups. The active intervention included a skill-building strategy based on Theory of Planned Behavior using Motivational Interviewing. It included a low sodium (LS) diet (2500mg sodium/day), lycopene supplementation daily (8 ounces [oz] of LS sodium tomato juice or 11.5 oz of LS V8 juice), and omega-3 fatty acid capsules (350mg EPA, 50 mg DHA/capsule) 3/meal with each meal. Placebo patients received generic instructions to follow a LS diet, 8 oz/day of fruit juice with no lycopene (e.g. cranberry juice) and capsules that contained soybean oil, but that looked like intervention capsules. Cox proportional hazards modeling was done to determine time to event of cardiac hospitalization or death based on intervention group. Models were adjusted for age, gender, NYHA class and HF medications.

Results:
The two groups were similar at baseline. The active intervention group had better event-free survival (Figure, p = 0.03) independent of covariates. Placebo patients were 2.2 (95% CI 1.025 - 4.584) times more likely to experience an event.

Conclusion:
A diet-based intervention aimed at the pathologic pathways underlying the 3 most common HF symptoms is successful in reducing cardiac rehospitalizations and mortality.
Abstract 15392: Radiation-Induced Coronary Microvascular Leakage is Mediated by Loss of Endothelial Tight Junction Protein Claudin-1
Authors: Nepali Sarmila , Xu Shirley , Sonkawade Swati D , Sharma Umesh C , Pokharel Saraswati
Published: June 2023

Background:
Ionizing radiation (IR) exposure to the heart during cancer therapy induces coronary microvascular leakage. Increased microvascular porosity allows inflammatory cell extravasation into the perivascular and myocardial interstitial space leading to fibrotic end organ damage.

Hypothesis:
Radiation-induced coronary microvascular leakage is mediated by a loss of a constitutively expressed tight-junction protein, claudin-1 (cldn1).

Methods:
We studied the function of cldn1 in human coronary vascular endothelial cells (HMVECs) and in mice using oligonucleotide-based cldn1 gene deletion. For the gain of function, we developed vascular-specific (VE-cadherin promotor-linked) claudin-1 overexpressing transgenic (TG) mice. We studied the effect of IR in cldn1 loss and gain of function models. Capillary permeability was assessed by FITC-dextran permeability and Evans blue dye injection,
in vitro
and
in vivo,
respectively
.
The extent of end-organ damage was examined by analyzing myocardial and perivascular fibrosis.

Results:
In HMVECs, cldn1 gene silencing (control, 1.0±0.34; siRNA, 0.026±0.03; p<0.001) led to increased permeability (control, 1.0±0.01, siRNA, 1.3±0.01; p<001). In mice, genetically engineered anti-cldn1 siRNA administration knocked down myocardial cldn1 expression (control, 1.0±0.18; siRNA, 0.51±0.11; p=0.04). Functionally, cldn1 knockdown led to increased microvascular extravasation (control, 1.0±0.23, cldn1-siRNA 0.38±0.27 p=0.002). Cardiac morphometry in these mice showed myocardial fibrosis (control, 1.0±0.19; siRNA, 1.29±0.35; p=0.048). Cardiac irradiation significantly decreased myocardial cldn1 (control 1.0±0.27, IR 0.45±0.31; p=0.001) and dye leakage (control 1.0±0.18, IR 1.28±0.22; p=0.007). Conversely, endothelium-targeted cldn1 overexpression in mice prevented abnormal microvasculature permeability (WT 1.0±0.23, TG 0.38±0.27 p=0.002) and conferred protection from IR-induced fibroinflammatory changes.

Conclusions:
Our study shows that cldn1 is crucial for maintaining coronary microvascular permeability. Prevention of cldn1 degradation or cldn1 reconstitution using novel pharmacotherapeutic approaches can prevent cardiac end-organ damage induced by cardiac IR.
Abstract 15400: A Call to Action for Mental Health Needs Amongst Patients Diagnosed With Critical Limb Ischemia: Insights From the Scope-Cli Registry
Authors: Smolderen Kim G , Shishehbor Mehdi H , Alabi Olamide , Fitridge Robert , Provance Jeremy A , Babrowski Trissa , Romain Gaelle , SOUKAS PETER A , Bunte Matthew C , Scierka Lindsey , van Roij Evy , Mena-Hurtado Carlos ,   On Behalf of the SCOPE-CLI Consortium
Published: June 2023

Introduction:
Late stage PAD, or critical limb ischemia, is a devastating diagnosis where patients deal with pain, non-healing wounds, high rates of major amputation, and increased cardiovascular event risk. Previously, we have demonstrated that depression and perceived stress are prevalent in early-stage PAD and associated with adverse health status and mortality risk, but it is unclear what the mental health burden is in patients with late stage disease.

Methods:
In the ongoing 11-center SCOPE-CLI registry, patients with a new diagnosis of CLI (Rutherford 4-6) were enrolled and underwent assessments with the 8-item Patient Health Questionnaire (PHQ-8, range 0-24, score ≥10 denoting positive screen, scores 10-14 moderate, 15-19 moderately-severe, and 20-24 severe depressive symptoms) and the 4-item Perceived Stress Scale (PSS-4, range 0-15, scores ≥6 exceed normative population scores).

Results:
Between 01/2020 and 06/2022, we enrolled 184 patients with mean ages of 67.5±10.4 years, 20.1% were black, and 70.1% was male. A total of 22.3%, 71.7%, and 6.0% had Rutherford 4, 5, and 6, respectively. A total of 31.8% had a positive depression screen (mean 7.4±6.2), with rates being equally high across Rutherford categories. Somatic symptoms (sleep, appetite, fatigue) were prominent along with cognitive symptoms (loss of interest, feeling hopeless). A total of 6.3% presented with severe depressive symptoms. Perceived stress levels were high (mean 4.8±3.5) with 41.0% exceeding the normative scores and highest stress rates observed amongst those with Rutherford 5-6 observed (37.8-43.4%). (
Figure
).

Conclusions:
As clinically relevant depressive symptoms and perceived stress are very common experiences in patients navigating a CLI diagnosis, integrated pathways that include behavioral evaluation and treatment in concert with CLI treatment goals are urgently indicated.
Abstract 15407: Benefits of the Sglt2 Inhibitor Empagliflozin on Epicardial Adipose Tissue in Non-Diabetic Hfref
Authors: Santosgallego Carlos G , Requena-Ibanez Juan Antonio , Picatoste Botija Maria Belen , Ishikawa Kiyotake , Sanz Javier , Fuster Valentin , Badimon Juan J
Published: June 2023

Introduction:
SGLT2 inhibitors cause weight loss, and reduce epicardial adipose tissue (EAT) on T2DM. The effects of T2DM on non-diabetic HFrEF remain unclear.

Methods:
HFrEF was induced in non-diabetic pigs via percutaneous oclusion of LAD (LVEF 30±4%). Animals were randomized to empagliflozin 10mg daily or placebo (n=8/group) for 2 months, and animals were subsequently euthanized. Cardiac MRI and echocardiography were performed before and after treatment. Invasive hemodynamics, histology and molecular biology were performed at 2-months. Primary culture of adipocytes from EAT was performed.

Results:
Empagliflozin caused reverse LV remodeling, reduced LVEDV and improved LVEF (Table). EAT volume was inversely associated with myocardial consumption of free fatty acids (r=-0.6, p=<0.05). Empagliflozin reduced EAT, both as MRI-determined EAT volume and histologically-evaluated adipocyte size (Table). Empagliflozin induced lipolysis in the adipocytes from EAT, as demonstrated by lower levels of triglycerides and perilipin (main protein responsible for triglyceride storage), and higher levels of the lipolytic enzymes lipoprotein lipase and hormone-sensitive lipase. Empagliflozin caused an anti-inflammatory effect on EAT as shown by lower levels of macrophage infiltration and MCP-1. Empagliflozin exerted antioxidant actions, as per reduced levels of 8-OH-deoxyguanosine and malonildealdehide (oxidative stress markers in nuclei and protein, respectively.

Conclusions:
Empagliflozin exerts multiple benefits on EAT in non-diabetic HFrEF. Empagliflozin induces a lipolytic state that reduces EAT volume, and it also ameliorates the oxidative and inflammatory status of EAT.
Abstract 15442: Pharmacological and Atrial Cardiomyocyte-Specific Inhibition of Tumor Necrosis Factor (TNF) Protects Against Adverse Atrial Remodeling and Valvular Atrial Fibrillation an a Mouse Model of Aortic Regurgitation
Authors: Lakin Robert , Liu Xueyan , CHEN WENLIANG , DEBI RYAN , Yakobov Simona , Polidovitch Nazari , Backx Peter
Published: June 2023

Introduction:
Atrial fibrillation (AF) is the most common sustained supraventricular arrhythmia worldwide. Most conditions linked to AF, such as cardiovascular (CVD) and valvular disease, are linked to elevated atrial pressures and atrial cardiomyocyte (CM) stretch, which are powerful stimuli for atrial remodeling. We have shown that the proinflammatory and mechanosensitive cytokine tumor necrosis factor (TNF) is a key mediator of stretch-related atrial remodeling and AF.

Hypothesis:
As TNF is a critical factor mediating fibrosis, hypertrophy, inflammation, and arrhythmias in CVD, we hypothesized that targeting TNF-dependent signaling may offer a novel therapeutic target in AF patients.

Methods:
We created an aortic regurgitation (AR) mouse model of volume overload characterized by elevated left ventricular end diastolic (LVEDPs) and atrial pressures. To examine TNF-dependent AF pathogenesis in AR, the impact of
Tnf
excision in atrial CMs (Cre-recombinase expression controlled by an
Nppa
promoter) and pharmacological TNF inhibition with Etanercept (twice-weekly, 2.5 mg/kg) beginning 2-days (early) or 1-week (delayed) post-AR in CD1 mice were examined. Cardiac structural, functional, and electrophysiological properties were assessed using echo, pressure telemetry, immunohistochemistry,
in vivo
intracardiacs, and
ex vivo
optical mapping of AF.

Results:
Four weeks of AR resulted in chronically elevated LVEDPs and progressive LV dilatation, functional impairment, and hypertrophy in the absence of ventricular arrhythmias with or without TNF inhibition compared to SHAM mice. In atria, AR resulted in both
in vivo
(8/11, 73%) and
ex vivo
(5/8, 63%) AF inducibility, which was associated with hypertrophy, fibrosis, and macrophage infiltration (F4/80
+
) as well as decreased conduction velocity and atrial effective refractory periods. Importantly, atrial CM-specific
Tnf
excision (n=10) and both early (n=8) and delayed (n=6) pharmacological TNF inhibition attenuated AR-induced adverse atrial remodeling and protected against AF inducibility.

Conclusions:
AF pathogenesis and vulnerability with AR requires TNF, suggesting TNF is an important therapeutic target for the prevention and treatment of AF linked to elevated atrial pressures.
Abstract 15468: Cd81 Interacts With Sodium Chloride Cotransporter in Lps-Induced Preeclampsia Rats and in Cultured Mouse Renal Distal Convoluted Tubule Cells
Authors: Wang Xiaoyan
Published: June 2023

Kidney regulates blood pressure mainly through salt/water reabsorption affected by sodium transport proteins along nephron. We have reported that placental CD81 plays an important role in the pathogenesis of preelampsia. In order to explore the connection between CD81 and renal sodium transport proteins in preelampsia, we studied their interactions in vivo and in vitro. On the gestation day 18 of lipopolysaccharide-induced preeclamsia model rats with hypertension and albuminuria, the renal protein abundances of NKCC2 (150±14,% of control, n=5-6/group, mean±SE, t-test, same as below), p-NKCC2(567±70%), NCC (254±41) and WNK4 (196±12) were increased together with an increase of CD81 in kidney (225±46) and urinary exosomes(396±100). Renal NCC (215±30) was elevated while NKCC2 tended to increase at 17000g pellet in LPS group. There was no change of both NKCC2 and NCC at 200,000g pellet. In kidney homogenates, CD81 was coimmunoprecipitated with NKCC2, NCC, not NHE3 while NKCC2, NCC were coimmunoprecipitated with CD81 not CD63. In renal plasma-membrane-enriched fraction NCC, not NKCC2, was also increased in preeclampsia rats. In the apical membrane of renal tubules CD81 was colocalized with NCC while minimum colocalization was found between CD81 and NKCC2. In cultured mouse distal convoluted tubule cells, CD81 was colocalized with NCC while CD81 deletion with siRNA decreased NCC and partially reversed lipopolysaccharide-induced increase of NCC. Furthermore, CD81 was detected only in the urinary exosomes from preeclampsia patients along with increases in NKCC2 (6063±811) and NCC(1085±191)relative to normal pregnant women (n=6/group, t-test). Those findings suggest that CD81 interacts with renal NCC in vivo and in vitro and the increase of renal CD81 may up-regulate NCC, possibly NKCC2 as well, and contribute to the increased blood pressure in preeclampsia.
Abstract 15512: The Hepatic Lipid Type and Content Had an Impact on Hepatic Inflammation and Fibrosis in Streptozotocin-Induced Diabetic Mice
Authors: Koseki Masahiro , Chang Jiuyang , Saga Ayami , Kanno Kotaro , Tanaka Katsunao , Omatsu Takashi , Okada Takeshi , Inui Hiroyasu , Ohama Tohru , Nishida Makoto , Sakata Yasushi
Published: June 2023

Introduction:
It has been reported that nonalcoholic fatty liver disease is an independent risk factor for cardiovascular diseases. We have recently demonstrated that diet-induced steatohepatitis was associated with cardiac dysfunction and those were improved by the selective PPARα modulator, pemafibrate in wild type mice.

Hypothesis:
The aim of the study is to characterize the relationship between hepatic lipid content and inflammation, induced by two type of diets in diabetic mice model and to test the effect of pemafibrate on those.

Methods:
C57BL/6 mice were injected with streptozotocin (STZ) (40mg/kg for 4 days). Mice whose ad libitum blood glucose were 200-350mg/dl, were used as diabetic mice. In 11-week-old STZ induced diabetic mice were administered wild high fat/high cholesterol diet (WD group) or high fat/high cholesterol/ high sucrose/bile acids diet (NASH group), with or without pemafibrate for 12 weeks. Lipid profiles, liver cytokines, hepatic mRNA, liver histology were investigated.

Results:
Total cholesterol and unesterified cholesterol were significantly more accumulated in the NASH group compared to the WD group. Triglyceride were not changed between two groups. F4/80 positive area and sirius-red positive area were significantly increased in the NASH group compared to the WD group. Inflammatory cytokines such as Tnfa, Il1b and fibrosis associated genes Col1a1 and Col1a2 were also significantly increased in the NASH group, suggesting the presence of active inflammation and fibrosis in the NASH group. By co-administration with pemafibrate, serum HDL-C level was increased by 48.7% and the non-HDL-C level was decreased by 31.4% in the NASH/pemafibrate group. Importantly, total cholesterol and unesterified cholesterol were markedly decreased in the NASH/pemafibrate group. Along with those, F4/80 positive area and sirius-red positive area were decreased in the NASH/pemafibrate group by more than 50%. Hepatic Tnfa expression was decreased by 56.3% and Tgfb, Col1a1, Col1a2 was significantly decreased.

Conclusions:
Our study demonstrated that more total cholesterol and unesterified accumulation induced hepatic inflammatory responses and fibrosis in the diabetic mice and pemafibrate had the beneficial impacts on those.
Abstract 15518: The Effects of High Glucose and Lipoprotein Lipase on Macrophage Regulatory Function
Authors: Bhatnagar Ashita , Alenghat Francis J
Published: June 2023

Introduction:
Atherosclerosis has an established inflammatory, macrophage(MФ)-mediated component which can be exacerbated by uncontrolled diabetes. However, there are also protective, regulatory MФ processes that can help quell atherosclerosis. Lipoprotein Lipase (LPL) on the MФ surface hydrolyzes circulating triglyceride-rich lipoproteins into glycerol and free fatty acids to provide a non-glucose energy source.

Hypothesis:
We hypothesized that regulatory MФs have high LPL activity, which could promote fatty acid metabolism to drive inflammation-resolving functions, and that this LPL activity is inhibited by ambient glucose.

Methods and Results:
MФs were cultured and differentiated from bone marrow harvested from wild-type mice aged 9-16 weeks. Two weeks after harvest, these MФs were polarized with (1) interferon-γ and lipopolysaccharide to stimulate classically-activated, inflammatory MФs or (2) interleukin-4 to stimulate alternatively-activated, regulatory MФs. The resulting phenotypes were validated by canonical transcript markers (Tnfa/Nos2 for inflammatory MФs and Ym1/Arg1 for regulatory MФs) and cell surface markers (CD86 and CD206, respectively). Using a triglyceride analog that fluoresces when cleaved by LPL, we found that regulatory MФs have far greater LPL activity than inflammatory macrophages at baseline (33 ± 2 vs 1 ± 0.2 a.u., p < 0.001, n=8). By varying glucose concentration during the polarization, this effect was most pronounced at normal, physiological concentrations of glucose (5 mM, ~90 mg/dL) and blunted at a high-glucose concentration that can be found in uncontrolled diabetes (25 mM). We also report on lipoxygenase expression and specialized pro-resolving mediator production as effector functions downstream of LPL activity in regulatory MФs.

Conclusions:
Taken together, our findings support a model in which regulatory MФs are programmed to utilize LPL to drive the inflammation-resolving phenotype and that this process works best at low, physiologic glucose concentrations. Future translational work will investigate whether a lack of LPL-mediated regulatory MФ function is a reason why patients with uncontrolled diabetes have higher atherosclerotic burden and plaque instability.
Abstract 15524: Temporal Changes in the Published Clinical Cardiology Evidence Base
Authors: Mai William , Jhawar Nikita , Xie Guozhan , Kusumoto Fred
Published: June 2023

Background and Objective:
The published cardiology literature is critical for scientific communication, and we sought to determine if the published clinical cardiology evidence base has evolved over time.

Methods:
We evaluated the cardiology related publications in the medical journals (M) JAMA (J) and the New England Journal of Medicine (N) and all publications by the specialty journals (CA) Circulation (Ci) and the European Heart Journal (E) in 2000, 2010, and 2020 and added 2001, 2011, and 2021 for JAMA to account for potential annual variability.

Results:
Publication information is shown in the graph. For the M journals there has been an increase in the percentage of randomized controlled trials (RCT). For the CA journals there has been a significant decrease in single center studies (Si) and a proportionate increase in reanalysis (Re) of data whether from subanalysis of an RCT, registry, or meta-analysis. For CA journals there has been an increase in the size of clinical studies mainly due to the use of larger datasets from registries or claims (Ci: 2000: 4,500/study; 2010: 9,298/study; 2020: 258,725/study and E: 2000: 2,282/study; 2010: 12,652/study; 2020: 249,388/study). CA journals have an increased of studies with patients from multiple continents (Ci: 2000: 5.5%; 2010: 13%; 2020: 35% and E: 2000: 7%; 2010: 14.8%; 2020: 18.8%). There has been an increase in the percentage of clinical studies with documentation of different populations by region or color (In 2020, 58% for all Ci articles and 52% for US and multicontinental EHJ articles), although the number of nonwhites remains small in most studies unless the study was specifically addressing disparities in care.

Conclusions:
Over the last two decades, the published evidence base has changed with more RCT in the Med journals and more studies that reanalyze large datasets in CA journals. Published studies are now larger, more likely to involve multiple continents, and document population details.
Abstract 15533: The Association Between Life’s Simple 7 and Arterial Stiffness in Black and White Women
Authors: Spikes Telisa , Pelkmans Jordan , Dunbar Sandra B , Igho-Pemu Priscilla E , Taylor Herman A , Mehta Puja , Quyyumi Arshed A
Published: June 2023

Background:
Black women have a higher prevalence of nonideal cardiovascular health (CVH) related risk factors across most components of the American Heart Association’s Life’s Simple 7 (LS7) than White women. Due to a higher burden of CVH-related risk factors, research suggests that Black women have accelerated and higher arterial stiffness than White women. However, the association between LS7 CVH and arterial stiffness among Black and White women is unknown.

Objective:
To examine the association between LS7 and arterial stiffness and investigate whether race modifies this association.

Methods:
We examined 1,640 adult women (n=1165 White, n=475 Black) from the Morehouse-Emory Center for Health Equity and Predictive Health Institute cohorts. Carotid-femoral pulse wave velocity (cfPWV), a marker of arterial stiffness, was assessed by applanation tonometry (SphygmoCor®). LS7 summary score (range 2-13) of 2-4 indicate ‘poor’, 5-9 ‘intermediate’, and 10-13 ‘ideal’ CVH. Cross-sectional analysis between the LS7 CVH 3-level categorical variable, cfPWV, and effect modification by race were examined. Multivariable linear regression models were conducted to examine the association between LS7 CVH and cfPWV, adjusting for model 1:(race, age); model 2:(model 1+ education, income).

Results:
Mean age 51±10.2 years. Compared to ‘ideal’ CVH, ‘poor’ (
β
=1.08 m/s; 95% CI, 0.44, 1.71) and ‘intermediate’ (
β
= 0.32 m/s; 95% CI, 0.20, 0.45) CVH groups had higher mean cfPWV in the minimal and fully adjusted models. Interaction of LS7 CVH and race on cfPWV was significant (p=.03) with Black women having higher mean cfPWV for each CVH group compared to White women. In race-stratified models, Black women with ‘poor’ (
β
=1.56 m/s; 95% CI, 0.79, 2.32) and ‘intermediate’ (
β
=0.46 m/s; 95% CI, 0.23, 0.68) CVH had higher cfPWV after minimal and full adjustment.

Conclusions:
Better CVH was associated with lower arterial stiffness. Compared to White women, Black women had greater arterial stiffness that was most pronounced for ‘poor’ and ‘intermediate’ CVH. Given that arterial stiffness is a prognostic measure, further study of health behaviors and psychosocial factors that contribute to higher arterial stiffness in Black women is warranted.