bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 14982: Impact of Health Literacy on Long-Term Outcomes in Patients with Heart Failure
Authors: Tsuchihashi-Makaya Miyuki , Matsuoka Shiho , Kayane Takahiro , Koizumi Masako , Yamada Michiyo , Okada Akiko , Yazawa Miyuki , Kinugawa Shintaro
Published: July 2021

Introduction:
Health literacy (HL) is an important concept for patient education and disease management of heart failure (HF). It has been demonstrated that HL is associated with clinical outcomes including death and readmission. Although previous research on HL has predominantly focused on functional HL (the ability to “read and write”), the World Health Organization advocates evaluation of comprehensive HL, including the ability to access information (communicative HL) and critically evaluate information (critical HL). However, the impact of these three HL components on long-term adverse outcomes in patients with HF has never been fully investigated.

Methods:
We conducted a prospective observational study to examine the associations between HL level and risk of death or readmission in 234 patients (mean age, 67.8 years; male, 62%). HL, including subscales of functional, communicative, and critical HL, was assessed using the HF specific HL scale, which has been reported to be valid and reliable in patients with HF. Sociodemographic, clinical, and depressive symptoms were also assessed. Mortality and hospitalization data were obtained during a 2 year follow-up (median duration, 707 days).

Results:
Among all patients, 19.7% exhibited a low HL score. In each subscale, 19.7%, 23.1%, and 23.9% of patients exhibited low functional, low communicative, and low critical HL, respectively. Patients with low HL were older and living alone. In multivariate Cox regression, low critical HL was independently associated with higher readmission exacerbated HF after controlling for demographic and clinical characteristics, HF severity, depressive symptoms at baseline (unadjusted rate, 23.2% vs. 9.1%; adjusted hazard ratio, 3.89 [95% CI, 1.24–12.21]; P=0.02). However, there was no association between all types of HL and mortality, and between functional and communicative HL and readmission due to HF.

Conclusions:
Critical HL is an independent risk factor for HF readmission in patients with HF. To improve clinical outcomes in patients with HF, effective interventions should be developed to improve patient skills for critically analyzing information and making decisions.
Abstract 150: Small Variations in Defibrillation Electrode Placement Affect Shock Success
Authors: Esibov Alexander , Melnick Sharon B , Chapman Fred W , Sullivan Joseph L , Walcott Gregory P
Published: July 2021

Background:
Defibrillation is essential in treating cardiac arrest patients with ventricular fibrillation (VF). The success of the shock in terminating VF depends on many previously studied factors, including waveform and energy dose. It has been observed clinically that defibrillation electrode placement varies widely. The goal of this study was to assess the effect of small differences in defibrillation electrode placement on first shock defibrillation success rates in swine.

Methods:
In 10 anesthetized swine weighing 32.7 ± 2.5 kg (mean ± standard deviation), electrode pads were attached at 3 different symmetric positions: the medial edge of each electrode was placed 3% (2.1 ± 0.1 cm), 7% (4.7 ± 0.1 cm), or 11% (7.5 ± 0.2 cm) around the circumference of the chest from mid-sternum (Fig. 1). Biphasic impedance-compensated shocks were delivered through the pads with a 50-ohm resistor added in series to replicate human impedance. The order of the 3 positions was block randomized for each experiment. At each electrode position, 24 episodes were run: VF was electrically induced and up to 2 shocks were delivered. Primary outcome was defined as successful VF termination after the first shock; secondary outcome was the cumulative success of both the first and second shock.

Results:
Across all experiments, at the 3%, 7%, and 11% positions, first shock success was 38.3% (92/240), 48.3% (116/240), and 36.7% (88/240) (p = 0.02, Chi-square Test); cumulative 2-shock success was 59.6% (143/240), 63.8% (153/240), and 55.4% (133/240) (p = 0.18), respectively. The lowest first shock success was at the 3% position in 6 of 10 animals, at the 11% position in 4 of 10 animals, and never at the 7% position.

Conclusions:
Small changes in electrode pad placement can significantly affect defibrillation shock success. Although anatomic differences may prevent the existence of one optimal electrode position, one position may be more effective for VF termination than another in a given patient.
Abstract 15015: Multimodal
in vivo
Computed Tomography and Optical Imaging for Simultaneous Functional, Metabolic and Molecular Myocardial Analysis
Authors: van Deel Elza D , Ridwan Yanto , Belenkov Sasha , Essers Jeroen
Published: July 2021

Introduction and Hypothesis:
The prevalence of myocardial infarction increases with the average age of the population and is currently the leading cause of death worldwide. In order to define anatomical changes and biomarkers related to cardiac infarction, we tested the use of combined computed tomography (CT) and near infrared fluorescent (NIRF) probes to facilitate non-invasive imaging of processes concerned with tissue degeneration/regeneration. This development of new non-invasive diagnostic methods will lead to better treatment options in the future.

Methods:
Mice were subjected to left anterior descending artery (LAD) ligation inducing an acute myocardial infarction and subsequently imaged
in vivo
using
fast
and
low dose
microCT scanning (QuantumFX, Perkin Elmer) and near infrared (NIRF) probes to monitor MMP activity (MMPsense680). Immediate contrast enhanced CT imaging using eXIA160 during its blood-pool phase allowed registration of changes in ventricular anatomy and function of important global parameters, like end-diastolic volume (EDV), end-systolic volume (ESV). These were used to calculate stroke volume (SV), and left ventricular ejection fraction (LVEF). Delayed eXIA160 contrast uptake during its myocardial phase, subsequently allowed analysis of the infarct size and infarct healing. In addition,
in vivo
molecular MMPsense localization was combined with micro-CT imaging for accurate 3D co-registration.

Results:
Changes in ventricular anatomy and myocardial viability were assessed 3 hours and 2 months post LAD occlusion in the same animal, demonstrating the feasibility of monitoring myocard viability over time. The decreased uptake of eXIA160 in the myocardium was subsequently quantitated. A concomitant increase in MMP activity, as determined by fluorescence mediated tomography using MMPsense680, could be localized to the infarcted site.

Conclusions:
Non-invasive imaging, using NIRF probes, enables longitudinal imaging of processes concerned with myocard infarction. Consequently, disease progression can be monitored over time and the effect of (new) pharmacotherapy can be studied.
Abstract 15036: Role of miR-195 in Aneurysm Formation
Authors: Zampetaki Anna , Attia Rizwab , Mayr Ursula , Gomes Renata , Phinikaridou Alkystis , Yin Xiaoke , Lnagley Sarah , Willeit Peter , Fanshawe Bruce , Fava Marika , Barallobre Barreiro Javier , So Po-Wah , Abbas Abeera , Jahangiri Marjan , Waltham Matthew , Botnar Rene , Smith Alberto , Mayr Manuel
Published: July 2021

Introduction:
Abdominal aortic aneurysm (AAA) is a degenerative disease of the aorta. Apart from the miR-29 family, miR-195 has been implicated in the regulation of the vascular extracellular matrix (ECM). Our aim was to assess the effect of the miR-15 family in aneurysm formation.

Methods and Results:
Expression of the miR-15 and miR-29 families was assessed in human aortic tissue. In human aneurysmal tissue, miR-195 expression was reduced to the same extent as miR-29b. In plasma, an inverse correlation of miR-195 was observed with the presence of AAA and aortic diameter. To evaluate the effect of miR-195 in ECM deposition, proteomics analysis was performed on the secretome of murine aortic smooth muscle cells (SMCs) following miRNA manipulation. MiR-195 targeted a cadre of ECM proteins, including collagens, proteoglycans, elastin and proteins associated with elastic microfibrils, albeit miR-29b showed a stronger effect. Systemic administration of cholesterol bound antagomiRs revealed better targeting efficacy of miR-195 compared to miR-29b in the uninjured aorta. A significant increase in aortic elastin expression was observed in wildtype animals injected with antagomiR-195. Yet, in apolipoprotein E-deficient mice receiving angiotensin II, inhibition of miR-29b but not miR-195 led to a significant inhibition of aortic aneurysm formation.

Conclusions:
We provide the first evidence that miR-195 may contribute to the pathogenesis of aneurysmal disease. Levels of miR-195 were reduced in human aneurysmal tissue and in plasma of patients with AAA. Although, miR-195 emerged as a potent regulator of aortic ECM, its inhibition was less effective than miR-29b in preventing aneurysm formation in preclinical models.
Abstract 15043: Circulating microRNA as Novel Biomarkers of Doxorubicin Induced Cardiotoxicity
Authors: D’Alessandra Yuri , Pontremoli Marta , Milano Giuseppina , Scopece Alessandro , Pompilio Giulio , Colombo Gualtiero I
Published: July 2021

Background:
Cardiotoxicity is a well-known side effect of Doxorubicin (Dox), leading to drug-induced heart dysfunction. Early prediction of disease onset is difficult, thus a new class of heart impairment biomarkers is needed. A role for miRNAs as circulating biomarkers of Dox-mediated heart dysfunction has never been tested.

Purpose:
To identify differentially expressed circulating miRNAs of early and late Dox-induced cardiotoxicity using an animal model.

Methods:
Healthy mice were administered 24 mg/Kg cumulative dose of Dox during a period of 2 weeks followed by 2-month recovery. Cardiotoxicity insurgence was detectable by echocardiography as early as 1 week after treatment. The expression of 367 miRNAs was analyzed in plasma samples from treated and untreated animals (n=6/group) at the end of the treatment (2 weeks) and 2 months post-administration (recovery phase) followed by singleplex qPCR validation of all miRNAs of interest.

Results:
We identified 5 Dox-regulated miRNAs of acute phase. MiR-34a, -34c, and -133a showed an upregulation (6.5, 14.9, and 15 folds, respectively, p≤0.01), whereas miR-16 and miR-451 were negatively regulated by the drug (-2.7 and -2.8 folds, respectively, p<0.05). We then investigated the expression of these miRNAs 2 months after the end of the treatment, but none showed a significant modulation. Conversely, miR-326 was positively regulated by Dox in the plasma of mice after 2 months of recovery (1.53 folds, p<0.05).

Conclusions:
This is the first study investigating the possible use of circulating miRNAs as biomarkers of both early and post-treatment Dox-induced cardiotoxicity. Our results revealed a strong regulation of 5 plasmatic miRNAs in response to early damage. Early responders returned to basal levels of expression 2 months after drug administration, indicating an acute damage induced release. This prompts for further studies in the clinical setting to test their suitability as toxicity markers beside cardiac troponin. By contrast, we could not identify strong markers of late damage, apart from a marginal difference in miR-326 circulating levels.
Abstract 15108: Imaging Coronary Atherosclerosis Using [18F]FDG PET-CT: Validation Study in Pigs
Authors: Tarkia Miikka , Saraste Antti , Stark Christoffer , Vähäsilta Tommi , Savunen Timo , Strandberg Marjatta , Saunavaara Virva , Tolvanen Tuula , Teuho Jarmo , Teräs Mika , Metsälä Olli , Rinne Petteri , Heinonen Ilkka , Sarja Nina , Pietilä Mikko , Saukko Pekka , Roivainen Anne , Knuuti Juhani
Published: July 2021

Introduction:
Positron emission tomography (PET) with 18F-fluorodeoxyglucose ([18F]FDG) can be used to detect atherosclerotic plaque inflammation. The degree of [18F]FDG uptake in different stages of coronary atherosclerosis remains largely unknown. Thus, we studied the amount of [18F]FDG uptake and feasibility of its in vivo quantification by combination of PET and computed tomography angiography (CTA) in a pig model of atherosclerosis.

Methods:
In order to induce coronary atherosclerosis, diabetes was caused by streptozotocin injections in farm pigs (n=10). After 6 months on high-fat diet, pigs underwent dual gated cardiac PET and CTA to measure [18F]FDG uptake in the proximal segments of coronary arteries as maximal target to background ratio (TBR = [18F]FDG uptake normalized to blood pool). Proximal coronary segments (n=33) were harvested for ex vivo measurement of radioactivity and for histology and measurement of tracer uptake into the vessel wall by autoradiography (ARG).

Results:
The pigs were hyperglycemic (12.3±4.7 mmol/L) and hypercholesterolemic (12.7±5.1 mmol/L) at the end of the study. The coronary arteries showed intimal thickening (n=16 segments) and atheroma (n=10 segments). Compared with the normal vessel wall, ARG showed 1.7±0.7 times higher accumulation of [18F]FDG in intimal thickening and 4.1±2.3 in atheroma (p=0.004 and p=0.003, respectively). Ex vivo mean vessel to blood ratio of segments with atheroma was higher than non-atherosclerotic segments (2.6±1.2 vs. 1.3±0.7, p=0.04). In vivo PET imaging showed the highest TBR of 2.7. However, maximal TBR was not significantly different in segments without atherosclerosis (1.1±0.5) and either intimal thickening (1.2±0.4, p=1.0) or atheroma (1.6±0.6, p=0.4) and no correlation was seen between the segmental TBR measured by in vivo PET-CT and [18F]FDG uptake measured by either biodistribution or autoradiography.

Conclusions:
We found increased [18F]FDG uptake in atherosclerotic lesions. However, quantification of [18F]FDG uptake in these relatively small and early stage atherosclerotic lesions by dual gated PET and CTA was not feasible. Further studies are needed to clarify feasibility of this approach in more advanced and highly inflamed atherosclerotic coronary plaques.
Abstract 15126: Cardiorespiratory Fitness and Atrial Fibrillation: A Population-based Follow-up Study
Authors: Kella Danesh K , Laukkanen Jari , Lloyd Michael S , Kuir Sudhir , Rauramaa Rainer , Butler Javed , Khan Hassan
Published: July 2021

Background:
Atrial fibrillation (AF) is the most common arrhythmia worldwide and has a complex association with exercise capacity. The relationship of atrial fibrillation with cardiorespiratory fitness (CRF) has not been previously investigated.

Methods:
Cardiorespiratory fitness, as assessed by maximal oxygen uptake (VO2max), was measured at baseline in 1,950 middle-aged men (mean 52.6 ± 5.1 years) from the Kuopio Ischaemic Heart Disease Study (KIHD).

Results:
During an average follow-up of 19.5 years, there were 305 incident AF cases (annual AF rate of 65.1/1000 person-years, 95% CI: 58.2 to 72.8). Overall a non-linear association was observed between CRF and incident atrial fibrillation. Rate of incident AF varied from 11.5 (95% CI: 9.4-14.0) for the first quartile of CRF, to 9.1 (95% CI: 7.4-11.2) for second quartile, 5.7 (95% CI: 4.4-7.4) for third quartile and 6.3 (95% CI: 5.0-8.0) for the fourth quartile respectively, Figure 1. Usual levels of CRF were non-linearly associated with AF risk, Figure 2. Age-adjusted HR comparing top versus bottom fourth of usual CRF levels was 0.67(95% CI: 0.48-0.95), which attenuated to 0.98 (95% CI: 0.66-1.43) upon further adjustment for several risk factors. These findings were comparable across age, body mass index, history of smoking, diabetes and cardiovascular disease status at baseline.

Conclusions:
Improved fitness as indicated by higher levels of CRF is protective of atrial fibrillation within an optimal range, beyond which the risk of atrial fibrillation rises again. Our findings are congruent with emerging data on the complex relationship between physical fitness and incident AF.
Abstract 15137: Vector Flow Mapping is a Novel Useful Technique in Assessing Inertia Force of Late Systolic Aortic Flow and Left Ventricular Early Diastolic Function
Authors: Wakami Kazuaki , Hachiya Kenta , Murai Syunsuke , Ito Tsuyoshi , Fujita Hiroshi , Goto Toshihiko , Fukuta Hidekatsu , Fukuta Hidekatsu , Tani Tomomitsu , Ohte Nobuyuki
Published: July 2021

Background:
We previously reported that the inertia force (IF) of blood flowing out of left ventricle (LV) during late-systole produces greater LV elastic recoil force and brings faster LV relaxation. Vector flow mapping (VFM
TM
, Hitachi-Aloka) enables us to see blood flow velocity vectors that are generated from conventional color Doppler imaging data at any phase of cardiac cycle without angle dependency. Using VFM, kinetic energy (KE) of ejecting blood flow during systole at the LV outflow tract (LVOT) can be obtained. Thus, we investigated whether the KE obtained at the LVOT during late systole (KE-ls) had any relations with the IF and invasively obtained LV function parameters.

Method:
Study subjects were 33 patients who underwent diagnostic cardiac catheterization and echocardiographic examination on the same day. Color Doppler images were acquired in the apical 3-chamber view. The frame rate ranged was from 40 to 51 frames per minute. Data analyses were performed offline using the commercially available software (DAS-RS1
TM,
Hitachi-Aloka). A data sampling area was set at the level just below the aortic valve in the LVOT. The KE-ls was computed as the sum of KE values computed in frame by frame basis during late-systole; late-systole was defined as the latter one-third of ejecting time. LV pressure wave was obtained using a catheter-tipped micromanometer, and then, the first derivative of LV pressure (dP/dt) and a time constant τ of LV pressure decay during isovolumic relaxation were calculated. From LV pressure-dP/dt relationships (phase loop), the IF was determined.

Results:
A significant positive correlation was observed between the KE-ls and the IF (r=0.79, p<0.0001). The log transformed KE-ls had significant correlations with both peak negative dP/dt (r=0.53, p<0.01) and the time constant τ (r=-0.67, p<0.0001).

Conclusion:
VFM is a new useful technique to see blood flow in the LV chamber. Noninvasively obtained KE-ls using VFM, which may be a noninvasive surrogate for the IF, has significant correlations with the parameters of LV relaxation.
Abstract 15162: Nanoparticle-Mediated Simultaneous Targeting to Mitochondria and Inflammatory Monocytes Confers Additive Cardioprotection Against Myocardial Ischemia-Reperfusion Injury
Authors: Ikeda Gentaro , Matoba Tetsuya , Nakano Kaku , Sunagawa Kenji , Egashira Kensuke
Published: July 2021

Backcround:
Targeting one mediator of myocardial ischemia-reperfusion (IR) injury failed to successfully reducing infarct size in clinical trials; therefore, an innovative approach may be to target more than 2 mediators at a time. Previously, we have engineered poly(lactic acid/glycolic acid) nanoparticle containing cyclosporine A (CsA-NP) and pitavastatin (Pitava-NP), and reported that the former inhibits the opening of mitochondrial permeability transition pore (mPTP) and the latter reduces monocyte-mediated inflammation in IR hearts. Here we tested the hypothesis that nanoparticle-mediated simultaneous targeting to mitochondria and monocytes confers additive and ultimate cardioprotection against IR injury.

Methods and Result:
In a murine model of myocardial IR injury, simultaneous treatment with CsA-NP and Pitava-NP at the time of reperfusion showed additive reduction in IR injury (infarct size) in wild-type mice (Fig. A). In cyclophilin D (CypD, a key regulatory molecule for mPTP opening)-KO mice, Pitava-NP reduced IR injury and recruitment of Ly6Chigh inflammatory monocytes whereas CsA-NP had no therapeutic effcts (Fig. B). In contrast, CsA-NP reduced IR injury in CCR2 (a receptor for monocyte chemoattractant protein-1)-KO mice. Flow cytometric analysis revealed that Pitava-NP, but not CsA-NP, inhibited the recruitment of Ly6Chigh inflammatory monocytes into IR heart. We then produced CypD/CCR2 -knockout mice and found that the double-KO mice displayed dramatic reduction in IR injury (Fig. C). Fluorescence molecular tomography showed that inflammation was markedly inhibited in CCR2-KO and CypD/CCR2-KO mice while residual inflammation was noted in CypD-KO mice (Fig. D).

Conclusions:
Nanoparticle-mediated simultaneous targeting to mitochondria and inflammatory monocytes can be developed as a novel therapeutic strategy that offers ultimately adequate cardioprotection in acute myocardial infarction and other clinical settings.
Abstract 15264: Clinical Outcomes and Cardiovascular Responses to Exercise Training in Heart Failure Patients with Preserved Ejection Fraction: A Systematic Review and Meta-Analysis
Authors: Dieberg Gudrun , Ismail Hashbullah , Giallauria Francesco , Smart Neil A
Published: July 2021

Background:
Exercise training induces physical adaptations for heart failure patients with systolic dysfunction but less is known about those patients with preserved ejection fraction.

Objectives:
To establish if exercise training produces changes in peak VO
2
and related measures, quality of life, general health and diastolic function in heart failure patients with preserved ejection fraction (HFpEF).

Methods:
We conducted a MEDLINE search (1985 to March 2014), for exercise based rehabilitation trials in heart failure, using search terms ‘exercise training, heart failure with preserved ejection fraction, heart failure with normal ejection fraction, peak VO
2
and diastolic heart dysfunction’. Seven intervention studies were included providing a total of 144 exercising subjects and 114 control subjects, a total of 258 participants.

Results:
Peak VO
2
increased by a mean difference (MD) 2.13 ml.kg
-1
.min
-1
(95% C.I. 1.54 to 2.71, p<0.00001) in exercise training versus sedentary control, equating to a 17% improvement from baseline. The corresponding data for V
E
/VCO
2
slope MD 0.85 ml.kg
-1
.min
-1
(95% C.I. 0.05 to 1.65, p=0.04); maximum heart rate MD 5.60 bpm (95% C.I. 3.95 to 7.25, p<0.00001); and 6 Minute Walk Test (6MWT) MD 32.1m (95% C.I. 17.2 to 47.1, p<0.0001); diastolic function; E/A ratio MD 0.07 (95% C.I. 0.02 to 0.12, p=0.005); E/E’ ratio MD -2.31 (95% C.I. -3.44 to -1.19, p<0.0001); Deceleration time (D
T
) MD -13.2 msec (95% C.I. -19.8 to -6.5, p=0.0001); Minnesota Living with Heart Failure Questionnaire (MLHFQ) MD -6.50 (95% C.I. -9.47 to -3.53, p<0.0001); Short Form (36) Health Survey MD 15.6 (95% C.I. 7.4 to 23.8, p=0.0002). In 3,744 hours patient-hours of training, not one death was directly attributable to exercise.

Conclusions:
Exercise training appears to effect several health-related improvements in people with heart failure and preserved ejection fraction.
Abstract 15335: Sex-Specific Cardiomyocyte Death in Heart Failure due to Sex-Specific Calcium Remodeling
Authors: Kararigas Georgios , Molina Cristina E , Summer Holger , Baczko Istvan , Golz Stefan , Regitz-Zagrosek Vera
Published: July 2021

L-type calcium (Ca2+) channels (LTCCs) play a key role in mitochondrial Ca2+ uptake, which may result in mitochondrial depolarization and metabolic collapse. Both apoptosis and necrosis occur in a sex-specific manner in heart failure (HF). However, the underlying mechanisms are unknown. We hypothesized significant sex differences in the cardiac transcriptome of end-stage non-ischemic dilated cardiomyopathy (DCM) patients. We compared left ventricular (LV) samples of age- and medication-matched male (
n
= 5) and female (
n
= 5) DCM patients with male (
n
= 10) and female (
n
= 8) controls by genome-wide expression profiling. Single gene differential analysis identified 4273 transcript clusters regulated in male, 1344 in female and 2817 in both male and female LVs (adjusted
P
< 0.01). Pathway analysis (adjusted
P
< 0.05) revealed induction of ECM-receptor interaction and several inflammatory pathways in male LVs, while oxidative phosphorylation and proteasome pathways were repressed. In female LVs, the Wnt and Hedgehog signaling pathways were induced, while the mTOR signaling pathway was repressed. Ca2+ signaling pathway was induced in both sexes. However, male LVs had a significantly higher induction of ca. 30% of genes encoding ion channels, accessory beta subunits and regulatory proteins than female LVs. Next, we isolated ventricular myocytes (VMs) from DCM patients and measured sarcoplasmic reticulum (SR) Ca2+ load and L-type Ca2+ current (ICa,L) by patch-clamp. While there were no sex differences in SR Ca2+ load, there was a 2-fold increase in ICa,L density in male vs. female VMs (
n
= 6-8 cells/group; each cell originated from a different individual;
P
< 0.05). The cytosolic release of cytochrome
c
was significantly higher in male than female LVs (
P
< 0.05). In view of that the anti-apoptotic protein BCL2 is involved in reducing recruitment of LTCCs, which in turn prevents mitochondrial Ca2+ overload, we found significantly higher BCL2 protein levels in female than male LVs (
P
< 0.05). Ca2+-mediated dephosphorylation of pyruvate dehydrogenase was higher in male than female LVs (
P
< 0.01) indicating more Ca2+ present in male mitochondria. We conclude that sex-specific regulation of Ca2+ homeostasis may account for sex differences in apoptosis and necrosis in HF.
Abstract 15348: Higher Mortality in Patients with Undiagnosed Hypertension: Is the Electronic Medical Record the Key to Their Identification?
Authors: Odunukan Olufunso W , Cha Stephen , Rahman Parvez A , Roellinger Daniel , Nyman Mark A
Published: July 2021

Introduction:
Patients with hypertension (HTN) may remain undiagnosed as the diagnosis requires systolic blood pressure (SBP) ≥ 140 mmHg or diastolic blood pressure (DBP) ≥ 90 mmHg on ≥ 2 different occasions.

Hypothesis:
We sought to evaluate outcomes of patients without diagnosed hypertension who have elevated BP meeting current thresholds for hypertension on 2 consecutive episodes.

Methods:
Together with retrospective billing and clinical data, electronic records of all outpatient visits in 2009 were used to identify 24,277 patients with 2 consecutive visits (2CHBP) with SBP ≥ 140mmHg or DBP ≥ 90 mmHg and were compared with a previously identified cohort of 53,998 known hypertension patients. Among the 2CHBP group, 6,071 (25%) were identified without a prior diagnosis of hypertension and were categorized as undiagnosed hypertension. They were compared with patients with known hypertension who had ≥ 1 visit in 2009. Cross-sectional analysis of cardiovascular comorbidities and proportional hazards for survival were done.

Results:
Of 5,978 undiagnosed HTN patients, 49% were female compared to 51% of 10,311 known HTN. In the undiagnosed HTN group, 19.5% were ≥ 80 years compared to 30.4% of known HTN patients. Cardiovascular comorbidities were generally lower in the undiagnosed HTN group compared to the known HTN group - heart failure (7% v 8%; OR 0.88, 95% CI: 0.77 - 0.99; p < 0.038), atrial fibrillation (11% v 12%, OR 0.88 95% CI 0.79 - 0.97) and coronary artery disease (17% v 26%, OR 0.57 95% CI 0.52 - 0.61). However after a four year followup, mortality was higher in the undiagnosed HTN group with unadjusted hazard ratio HR 1.4 (95% CI 1.29 - 1.60, p<0.0001) and HR 2.41 (2.16 -2.68, p<0.0001) when adjusted for age, sex, heart failure and diabetes.

Conclusions:
Over a quarter of patients meeting current thresholds on 2 consecutive visits did not carry a known diagnosis of hypertension. Although younger and with less cardiovascular comorbidities than those with diagnosed hypertension, mortality after 4 years was significantly higher in those with undiagnosed hypertension. Electronic records can be used to identify these high risk patients.
Abstract 15378: Mitochondrial Fission Plays a Crucial Role in Cardiomyocyte Adaptation to Energy Stress via Mitophagy
Authors: Ikeda Yoshiyuki , Maejima Yasuhiro , Ohishi Mitsuru , Sadoshima Junichi
Published: July 2021

Mitochondria are dynamic organelles that undergo fusion and fission. This study aims to clarify the role of mitochondrial dynamics on energetic stress resistance in cardiomyocytes (CMs).

Dynamin-related protein 1 (Drp-1) which mediates mitochondrial fission was localized primarily in the cytosol in CMs. Glucose deprivation (GD) induced modest mitochondrial accumulation of Drp1 in CMs within 4 hours. Four-hour GD increased the proportion of CMs with fused mitochondria (23.5±6.9 to 35.0±10.8%, p<0.05), but also increased that of CMs with fission (2.1±0.3 to 15.5±4.8%, p<0.05) in Ad-shScr-transduced CMs, whereas mitochondria with fission did not increase after GD in Ad-shDrp1-transduced CMs (0.3±0.6 to 0.8±0.5%). These results suggest that GD induces both mitochondrial fusion and fission, and Drp1 plays an essential role in mitochondrial fission in response to GD. Transduction with Ad-shDrp1 significantly increased TUNEL-positive CMs and reduced cell viability after 4 hours of GD compared to transduction with Ad-shScr, suggesting that endogenous Drp1 protects CMs against cell death during GD. We evaluated the role of Drp1 in mediating mitophagy using mitochondria-targeted Keima fluorescence. Keima has a bimodal excitation spectrum peaking at 440 and 560 nm, corresponding to the neutral and acidic pH states, respectively. Puncta with high 560/440, indicating the presence of mitochondria in lysosomes, were significantly increased after 4 hours of GD in CMs transduced with Ad-shScr, but not in CMs transduced with Ad-shDrp, suggesting that Drp1 is necessary for stimulating autophagic mitochondrial degradation. Next, we examined the role of fission on energy stress in vivo using cardiac specific Drp1 hetero knockout (Drp1-hetCKO) mouse. Translocation of Drp1 from cytosol to mitochondria significantly increased in response to 48-hour fasting in control mice but not in Drp1-hetCKO mice. Left ventricular ejection fraction did not change after fasting in control mice (79.8±6.4 to 78.6±8.3%). However that in Drp1-hetCKO mice decreased after GD (79.4±9.7 to 48.3±11.6%, p<0.05), suggesting that Drp1 acts to preserve cardiac function during fasting.

In conclusion, mitochondrial fission is crucial to adapt CMs to energy stress through mitophagy.
Abstract 15426: CSL112 Enhances Cholesterol Efflux Equally in Patients with High and Low High Density Lipoprotein Functionality
Authors: Gille Andreas , Hartel Gunter , D’Andrea Denise , Wright Samuel D
Published: July 2021

Introduction:
The ability of high-density lipoprotein (HDL) to remove cholesterol from macrophages in atherosclerotic plaque is thought to underlie its inverse correlation with cardiovascular risk. CSL112 is a novel formulation of apoA-I purified from human plasma and reconstituted to form HDL particles suitable for infusion. Two phase 1 studies in healthy subjects and one phase 2a study in coronary heart disease (CHD) patients have demonstrated favorable safety, pharmacokinetics and biomarker responses to single or multiple infusions of CSL112. The aim of the current study was to assess the influence of subject-specific HDL functionality, sometimes described as HDL dysfunction, on the pharmacodynamic effects of CSL112.

Methods and Results:
To assess HDL functionality, we measured the specific cholesterol efflux activity of apoA-I (cholesterol efflux divided by apoA-I) in subjects at baseline. CHD patients (mean ± SD = 7.3 ± 0.3 %efflux per g/L apoA-I (n=43)) had lower activity (p<0.001) than healthy subjects (mean ± SD = 8.8 ± 0.2 % efflux per g/L apoA-I (n=93)), consistent with prior work showing impaired HDL function in CHD patients. Every subject infused with CSL112 responded, showing increased apoA-I plasma levels and serum cholesterol efflux. To assess changes in HDL functionality upon infusion of CSL112, we plotted changes in cholesterol efflux
vs
changes in apoA-I. To account for variations of these analytes over time and dose, we plotted area under curve (AUC) for 0 to 24 h after infusion of various doses of CSL112. In all three studies, AUC for apoA-I correlated with the AUC for cholesterol efflux, with coincident lines, indicating equal elevation in efflux among healthy and CHD patients. To further examine this relationship, subjects were stratified into tertiles based on specific activity. Infusion of CSL112 elevated efflux similarly in subjects in the highest HDL functionality and lowest HDL functionality tertiles.

Conclusion:
ApoA-I from CHD patients compared to healthy subjects had lower specific activity in regards to cholesterol efflux. CSL112 causes strong elevation in cholesterol efflux regardless of HDL function and represents a promising candidate to rapidly remove plaque cholesterol and reduce recurrent atherothrombotic events.
Abstract 15445: The Sirtuin SIRT3 Blocks Doxorubicin-induced Cardiac Hypertrophic Response of Mice
Authors: Pillai Vinodkumar , Samant Sadhana , Sundaresan Nagalingam , Kim Gene , Gupta Mahesh P
Published: July 2021

Background and objective:
Doxorubicin is a chemotherapeutic drug widely used to treat variety of cancers. One of the serious side effects of doxorubicin is its toxicity to the heart. Previously, we have shown that overexpression of SIRT3 blocks the hypertrophic response of the heart to agonist treatments. This study was undertaken to investigate whether SIRT3 can also attenuate the doxorubicin-induced cardiac hypertrophic response in mice.

Methods and results:
Neonatal rat cardiomyocytes overexpressed with SIRT3 and treated with doxorubicin (10μM) showed 28% reduced mean fluorescent intensity for CM-H
2
DCFDA dye, compared to mock infected control cells treated with doxorubicin, thus suggesting that SIRT3 was capable of blocking doxorubicin-induced ROS synthesis in cardiomyocytes. To examine the cardioprotective effects of SIRT3 in doxorubicin-induced cardiotoxicity
in vivo
; we used a cumulative dose of 15mg/kg of doxorubicin for two different time points. One group of mice was treated intraperitoneally with 5mg/kg doxorubicin or an equal volume of saline every two weeks for a total of three doses. Transgenic mice having cardiac specific expression of SIRT3 (SIRT3-Tg) showed 33% reduced HW/BW ratio compared to control mice. Echocardiographic evaluation of hearts showed significantly reduced fractional shortening in control mice, compared to SIRT3-Tg mice (24.6 vs 34.7 %, P<0.05). SIRT3-Tg mice also showed significantly reduced fetal gene expression for ANF, βMHC and collagen-1 as determined by RT-PCR. Masson’s trichrome staining showed significantly reduced fibrosis in doxorubicin treated SIRT3-Tg mice compared to its control. Furthermore, electron microscopic analysis showed preserved mitochondrial and sarcomeres structures in doxorubicin treated SIRT3-Tg hearts, whereas in wild-type hearts these structures were highly disorganized. Second group of mice that received 15mg/kg dose for two weeks also showed similar results. Contrary to this, whole body SIRT3 knockout mice showed exacerbated cardiac hypertrophic response compared to wild-type mice in response to doxorubicin treatment.

Conclusion:
These results demonstrated that SIRT3 is an endogenous negative regulator of doxorubicin-induced cardiac hypertrophic response.
Abstract 15453: CaMKII Plays a Critical Role in Pressure Overload-Induced Left Ventricular Hypertrophy by Modulating Foxp1 Expression
Authors: Zhao Lin , Cheng Guangming , Jin Runming , Chen Lei , Samanta Anweshan , Girgis Magdy , Elias Harold K , Davani Arash , Choksi Kashyap , Vincent Robert , Yang Yanjuan , Hauptman Jeryl , Dawn Buddhadeb
Published: July 2021

Introduction:
The necessity of IL-6 signaling in pressure overload-induced left ventricular hypertrophy remains controversial. Whether Calcium/calmodulin-dependent kinase II (CaMKII) participates in IL-6-induced and pressure overload-induced hypertrophic signaling remains unknown.

Hypothesis:
CaMKII acts as a nodal point for integrating IL-6 signaling in pressure overload induced hypertrophic molecular pathways.

Methods and Results:
In vitro, exposure to IL-6 for 24 h induced hypertrophy in H9c2 cells. This hypertrophic response was associated with robust increase in phospho-CaMKII levels in IL-6-treated cells, and accompanied by increase in myocyte hypertrophy markers. Interestingly, Mef2a levels also increased, and this was associated with reduced Foxp1 expression, which has Mef2a specific binding site at CTAAAAATAG (-401 ~ -391) in Foxp1 promoter sequence. These effects were inhibited by selective CaMKII inhibitor KN-62. Next, we performed transverse aortic constriction (TAC) in IL-6-/- mice and wild-type (WT) C57BL/6J mice to induce hypertrophy. After 6 wks, TAC-induced hypertrophy was significantly attenuated in IL-6-/- mice compared with WT mice, documented by necropsy findings, echocardiography, and LV morphometry. Phospho-CaMKII levels increased significantly in control mouse hearts at 2 wks after TAC, while this increase was significantly blunted in IL-6-/- hearts (Figure). Moreover, and consistent with attenuated CaMKII activity, the expression of Mef2a was lower and expression of Foxp1 was higher in IL-6-/- hearts after TAC compared with controls.

Conclusions:
We identify a novel signaling module whereby regulation of Foxp1 by CaMKII plays a critical role in pressure overload-induced and IL-6-mediated cardiac hypertrophy. These molecular insights may lead to formulation of novel therapeutic agents for LV hypertrophy and dysfunction.
Abstract 15526: Predictors of Elective Support Device Insertion Prior to High Risk Percutaneous Coronary Intervention: Changing Trends between 1993-2013
Authors: Garg Aatish , Parashar Akhil , Agarwal Shikhar , Tuzcu E. Murat , Kapadia Samir
Published: July 2021

Introduction:
High risk percutaneous coronary interventions (PCI) may be preceded by elective support device insertion, though the evidence for such a practice is relatively weak. We sought to analyze the changing trends and predictors for prophylactic support device insertion in these patients over the last 20 years.

Methods:
Retrospective analysis of all patients who underwent PCI at Cleveland Clinic between 1993 - 2013 with or without support device insertion was performed. Trends for device insertion in high risk patients undergoing PCI, like those in clinical shock, acute myocardial infarction (MI) and unprotected left main intervention over the last 20 years were reviewed. Predictors for support device insertion in these patients from 1993-2000 were compared to those from 2001-2013.

Results:
A total of 666 elective support devices were inserted prior to high risk PCI from 1993-2013. Between 1993-2000, acute myocardial infarction (MI) (OR: 10.05), ejection fraction < 40% (OR: 8.31) and unprotected left main intervention (OR: 4.59) were the three most significant predictors for device insertion prophylactically. Between 2001-2013, unprotected left main intervention (OR: 9.91), ejection fraction < 40% (OR: 5.61) and clinical shock (OR: 5.24) were the biggest predictors, with the odds of support device insertion in acute MI in absence of clinical shock reduced to 1.96.

Conclusions:
There has been a significant decrease in the proportion of high risk PCIs with elective support device insertion, especially in those performed in patients with acute myocardial infarction in absence of shock.
Abstract 15529: Transgenic Short QT Syndrome Type 1 Rabbits (HERG-N588K) Mimic the Human Phenotype with Shortened Cardiac Repolarization, Ventricular Fibrillation, and Sudden Death
Authors: Odening Katja E , Franke Gerlind , Rieke Raphaela , Topf Aguiar de Medeiros Anna , Bodi Ilona , Perez-Feliz Stefanie , Pantulu Naga Deepa , Lang Corinna N , Ziupa David , Zehender Manfred , zur Hausen Axel , Bode Christoph , Jolivet Genevieve , Brunner Michael
Published: July 2021

Introduction and Hypothesis:
Short QT syndrome is an inherited ion channelopathy with an accelerated cardiac repolarization due to gain-of-function mutations in K+ channel genes. Patients are prone to ventricular tachycardia and sudden cardiac death (SCD). Detailed knowledge about mechanisms of arrhythmogenesis is lacking. We aimed at generating transgenic SQT1 rabbits as novel tool to investigate these mechanisms.

Methods:
FLAG-HA-tagged HERG-N588K cDNA and the b-MyHC promoter were integrated into a pBSII SK vector and microinjected into rabbit oocytes. SQT1 rabbits and their wildtype (WT) littermates were subjected to in vivo ECG and ex vivo monophasic action potential (AP) measurements. Electrical remodeling was investigated using real-time PCR.

Results:
We successfully generated 3 transgenic SQT1 founder rabbits. One of the founders died suddenly at the age of 60 days. In SQT1 rabbits (F1) not subjected to any experiments 3/11 died around day 40 vs. 0/8 WT rabbits (p<0.05). SQT1 founder rabbits and their offspring demonstrated a shortened QT interval compared to WT (QT, ms, SQT1, n=44, 147.8±1.7 vs. WT, n=24, 166.4±2.9, p<0.0001, heart-rate corrected QTi, %, 91.6±0.7 vs. 98.1±1.1, p<0.0001) with no gender differences among SQT1 rabbits. Moreover, in SQT1, AP duration was shortened in all segments of LV base, mid, and apex (LV base-lat, APD90, ms, SQT1, n=7, 118.6±5.1 vs. WT, n=9, 154.4±2.2, p<0.0001; APD75, 93.1±6.8 vs. 127.1±1.9, p<0.0001). Ventricular fibrillation (VF) occurred more often during AP measurements in SQT1 than in WT rabbits (SQT1, 5/13 vs. WT, 0/8, p<0.05).

FLAG-HA-HERG-N588K channels expressed in HEK cells demonstrated increased IKr steady currents and impaired inactivation indicative of a gain-of-function. Remodeling of ion channels and Ca2+ handling proteins occurred in SQT1 rabbits with a downregulation of L-type Ca2+ channel mRNA in LV septum-mid (SQT1, n=4, 0.84±0.1 vs. WT, n=6, 1.03±0.1, p<0.05) and of RyR2 in LV septum-apex (0.85±0.1 vs. 1.08±0.1, p<0.05).

Conclusion:
These first transgenic SQT1 rabbits over-expressing a HERG-N588K gain-of-function mutation mimic the human phenotype with shortening of QT and APD, a high rate of VF and SCD. These models are thus a useful tool to explore arrhythmogenic mechanisms in SQTS.
Abstract 15532: Altered Ubiquitination and Stability of Protein Inhibitor of Neuronal Nitric Oxide Synthase in the Paraventricular Nucleus of Chronic Heart Failure Rats: Role of Angiotensin II
Authors: Sharma Neeru , Liu Xuefei , Zheng Hong , Patel Kaushik
Published: July 2021

Introduction and Hypothesis:
Expression of neuronal nitric oxide synthase (nNOS) is decreased in the paraventricular nucleus (PVN) of rats with chronic heart failure (CHF), however the underlying molecular mechanism/s remain unclear. Recently, we demonstrated, Angiotensin II (Ang II) mediated increase in PIN: protein inhibitor of nNOS (0.76±0.10 Sham vs 1.12±0.09* CHF) which is known to down-regulate nNOS through disruption of active dimers (~60% decrease in dimer/monomer ratio) in the PVN of rats with CHF. Functionally impeded monomeric enzyme is degraded by ubiquitin proteasome system. Interestingly, PIN transcript levels remain unchanged in the PVN in CHF (1.00±0.23 Sham vs. 1.1±0.28 CHF). This observation prompted us to elucidate the molecular mechanism for the accumulation of PIN post-transcriptionally in the PVN in CHF

Methods and Results:
We used coronary artery ligation model of CHF in rats (6-8 weeks past ligation) and neuronal NG108-15 hybrid cell line. PIN translation was inhibited using cyclohexamide (CHX) for 0-4h after 20h of pretreatment with Ang II in NG108 cells. CHX mediated decrease in PIN expression was ameliorated with Ang II (0.19±0.04 vs 0.41±0.06* 4h). Proteasome inhibitor lactacystin (LC) treatment dramatically elevates PIN level suggesting the involvement of proteasome system in PIN regulation. Immunoprecipitation with ubiquitin antibody showed decrease PIN-Ub conjugates in Ang II-treated cells (1.04 ± 0.05 LC vs. 0.62 ± 0.07* LC AngII). In vitro ubiquitination assay in cells transfected with pCMV-(HA-Ub)8 vector revealed reduction of HA-Ub-PIN conjugates after Ang II treatment (9.2 ± 2.2 LC vs. 4.5 ± 0.6* LC Ang II). Furthermore, there was decreased accumulation of PIN-Ub conjugates in the PVN of CHF rats compared to Sham as revealed by immunohistochemistry.

Conclusions:
Taken together, our studies revealed that PIN is targeted for rapid degradation by the ubiquitin-proteasome pathway and Ang II delays the rate of degradation resulting in accumulation of PIN. We conclude that post-translational accumulation of PIN, mediated by Ang II, leads to a decrease in the dimeric active form of nNOS as well as protein levels of nNOS, which may lead to reduced nitric oxide resulting in over-activation of sympathetic drive during CHF.
Abstract 15546: Loss of Mitochondrial UCP3-Cytochrome c Oxidase Complexes Underlies Maladaptive Autophagy and Cardiac Failure in Anthracycline Cardiotoxicity
Authors: Dhingra Rimpy , Margulets Victoria , Jassal Davindar , Dorn Gerald , Kirshenbaum Lorrie A
Published: July 2021

Doxorubicin is known for its cardiotoxic effects and inducing cardiac failure. Herein we demonstrate a novel signaling pathway that functionally links activation and preferential mitochondrial targeting of Bnip3 to doxorubicin cardiotoxicity. Perturbations to mitochondria including increased calcium, ROS, loss of αΨm and mPTP opening were observed in cardiac myocytes treated with doxorubicin. This coincided with a decline in maximal respiratory capacity, loss of respiratory chain complexes of uncoupling protein 3 (UCP3) and cytochrome c oxidase complex IV subunit 1, (COX) and cell viability. Impaired mitochondrial function was accompanied by an accumulated increase in autophagosomes and necrosis demonstrated by increase release of LDH, cTnT and loss of nuclear High Mobility Group Protein 1 (HMGB-1) immunoreactivity. Interestingly, pharmacological or genetic inhibition of autophagy with 3-methyl adenine (3-MA), or Atg7 knock-down suppressed necrotic cell death induced by doxorubicin. Conversely, loss of function of Bnip3 or mutations of Bnip3 defective for mitochondrial targeting restored UCP3-COX complexes, mitochondrial respiratory integrity and suppressed necrotic cell death induced by doxorubicin. Finally, mice germ-line deficient for Bnip3 were resistant to the cytotoxic effects of doxorubicin displaying mitochondrial morphology, cardiac function and survival rates comparable to vehicle treated control mice. To our knowledge the findings of the present study provide the first direct evidence that doxorubicin triggers maladaptive autophagy and necrotic cell death of ventricular myocytes by a mechanism mutually dependent and obligatorily linked to Bnip3. Hence, therapeutic interventions to selectively inhibit Bnip3 may prove beneficial in suppressing mitochondrial injury and heart failure in cancer patients undergoing doxorubicin treatment.