bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 13771: Cardiopulmonary Exercise Testing is Predictive of Return to Work in Cardiac Patients After Rehabilitation
Authors: Voller Heinz , Salzwedel Annett , Reibis Rona , Kaminski Stefan , Buhlert Hermann , Eichler Sarah , Wegscheider Karl
Published: July 2021

Introduction:
Cardiopulmonary exercise testing (CPX) has an independent prognostic value, especially in cardiovascular patients. We aimed to evaluate parameters of CPX as predictors for return to work (RTW) at discharge of cardiac rehabilitation (CR).

Hypothesis:
Besides psychosocial as well as clinical parameters CPX variables are independent predictors of return to work in patients with cardiovascular diseases.

Methods:
We analysed sociodemographic and clinical data from a prospective registry of 489 patients (mean age 51.5±6.9 years, 87.9% men), who were referred to shortterm (3 weeks) inpatient CR between 06/2009 to 12/2011, predominantly after PCI (62.6%), CABG (17.2%) and heart valve replacement (9.0%). At admission, patients underwent noninvasive cardiac diagnostic (2D echo, exercise ECG, 6MWT) and a psychodiagnostic screening (HADS). CPX was performed at discharge for defining fitness.

Results:
During a mean follow up of 26.5±11.9 months 373 (76.3%) patients returned to work, 116 (23.7%) did not and 60 (12.3%) were retired. A higher number of comorbidities (p=0.011) and heavy work (p<0.001) were negatively associated with RTW whereas a higher exercise capacity at entry of CR (p<0.001) and elective PCI (p=0.02) increased the probability of RTW. After adjustment for covariates, max. work load (Watt) at CPX termination and the VE/VCO2-slope had an independent prognostic significance for RTW. A higher work load increased (p=0.009) while a higher VE/VCO2-slope decreased (p=0.027) the probability of RTW. Even for retirement, CPX had a prognostic value: the likelihood of retirement was smaller with increasing VO2AT (p=0.016).

Conclusion:
CPX is a meaningful tool to assess patients’ ability for return to work. Therefore it should be an essential part of functional assessment in CR for predicting participation in employment during two years after CR.
Abstract 13780: JNK Inhibition Mimics the Beneficial Effect of Roux-en-Y Gastric Bypass Surgery on Obesity-Induced Endothelial Dysfunction
Authors: Doytcheva Petia , Osto Elena , Baechler Thomas , Bueter Marco , Matter Christian M , Lutz Thomas A , Luscher Thomas F
Published: July 2021

Introduction:
Roux-en-Y gastric bypass surgery (RYGB) reduces weight and long-term cardiovascular risk in obese patients. We previously demonstrated that in diet-induced obese rats RYGB improves aortic endothelial vasorelaxation 8 days after surgery in a weight loss-independent manner. Improved endothelial function was associated with decreased JNK phosphorylation and increased Akt and eNOS phosphorylation in the aorta.

Hypothesis:
We investigated whether in vivo inhibition of JNK activity in sham-operated ad libitum-fed rats mimics these endothelial effects of RYGB.

Methods:
Male Wistar rats were fed a high fat (60%) high cholesterol (1.25%) diet for 7 weeks before undergoing either RYGB or sham surgery; sham-operated rats received vehicle (sham_AL) or the JNK-specific inhibitor SP600125 40mg/kg/day s.c. for 8 days post-surgery (sham_SP). Then, thoracic aortic rings were isolated and subjected to ex vivo isometric tension recordings. After submaximal contraction with norepinephrine (10-6mol/L), cumulative relaxation responses were performed to GLP-1 (7-36) amide (10-12 to 10-6mol/L) or insulin (10-12 to 10-5mol/L); tests were repeated after preincubation with the eNOS inhibitor L-NAME (10-4mol/L). Western blot analysis of JNK, Akt and eNOS was also performed on aortic tissue lysates.

Results:
Body weight did not differ between sham_SP and sham_AL rats, while the weight loss of RYGB rats was significant 8 days after surgery. GLP-1- and insulin-induced vasorelaxation responses improved in RYGB compared to sham_AL rats; interestingly, sham_SP rats had endothelial relaxation similar to RYGB rats. All responses were blocked by preincubation with L-NAME, suggesting an eNOS-dependent effect. JNK protein phosphorylation in aortic lysates was decreased in RYGB and sham_SP rats compared to sham_AL, while Akt phosphorylation was increased. eNOS phosphorylation and dimerization were also increased in RYGB and sham_SP compared to sham_AL rats.

Conclusion:
JNK inhibition seems to mimic the immediate eNOS-mediated endothelial protective effects of RYGB. Further, our study underlines a crucial role of JNK in obesity-associated endothelial dysfunction, suggesting a novel mechanism for the cardiovascular effects of RYGB.
Abstract 13781: Assessment of Microvascular Dysfunction by Transthoracic Echocardiography Compared to Positron Emission Tomography
Authors: Michelsen Marie M , Mygind Naja D , Pena Adam A , Olsen Rasmus H , Høst Nis , Christensen Thomas E , Hasbak Philip , Kjaer Andreas , Kastrup Jens , Prescott Eva
Published: July 2021

Introduction:
Microvessel dysfunction can be assessed by measurement of Coronary Flow Reserve (CFR) by Transthoracic Doppler Echocardiography (TTDE). Positron Emission Tomography (PET) is regarded as the gold standard for CFR measurements. Only one previous study based on 10 healthy males has validated TTDE against PET measured CFR. The aim of the study was to compare CFR measurements by TTDE to those obtained by PET in women with angina pectoris but no obstructive coronary artery disease (CAD).

Methods:
Patients (n=93) were randomly selected from a cohort of women with angina pectoris but no obstructive CAD and a successful CFR measurement by TTDE of the left anterior descending artery (LAD) during dipyridamole infusion (0.84 mg/kg). CFR was measured by rubidium-82 PET during adenosine infusion (0.84 mg/kg). A Quality Index (QI) of CFR measured by TTDE was assessed on a scale from 1-9 on the basis of visibility of vessel and characteristics of flow curves. Concordance between the methods was assessed by the Bland Altman method.

Results:
CFR was successfully measured in 88 women by rubidium PET (exclusion due to inability to complete examination (n=4) and one outlier with un-physiological high CFR value of 5.9). Median CFR (iq range) was 2.69 (2.24; 3.05) for PET and 2.36 (1.92; 2.75) for TTDE. CFR was systematically assessed higher by PET (mean difference 0.39, p<0.01. Reproducibility was compared to a large study (n=125) regarding PET-PET agreement (table 1).

Conclusions:
Results are similar to previous assessments of reproducibility of CFR assessed by PET. For patients with TTDE CFR QI > 6 there is a higher concordance between TTDE and PET CFR, indicating that the validity of CFR is dependent on quality. However, part of the divergence between PET and TTDE could be due to difference in method; TTDE estimates LAD flow velocities whereas PET estimates global flow per myocardial tissue mass. CFR measured with TTDE is a valid method to investigate cardiac microvascular dysfunction.
Abstract 13796: Quantitative Measurement of Cross sectional Area of Small Pulmonary Vessels Using High-resolution Ct in Chronic Thromboembolic Pulmonary Hypertension
Authors: Sugiura Toshihiko , Tanabe Nobuhiro , Kawata Naoko , Matsuura Yukiko , Kasai Hajime , Shigeta Ayako , Sakao Seiichiro , Kasahara Yasunori , Tatsumi Kouichiro
Published: July 2021

Background:
It is generally recognized that there is narrowing and diminution of the small vessels on pulmonary angiography in patients with chronic thromboembolic pulmonary hypertension (CTEPH). Previous studies have shown that these vascular alterations could be assessed by the cross-sectional area of small pulmonary vessels (CSA) on high-resolution CT (HRCT) in patients with COPD. However, it is unknown whether there is a correlation between CSA and hemodynamics in patients with CTEPH.

Aims:
We retrospectively evaluated the relationship between the CSA on HRCT and hemodynamics measured by right heart catheterization (RHC).

Materials and Methods:
61 patients (61±12 yrs, 13 male) with proven CTEPH underwent non-enhanced 64-slice HRCT (Aquilion ONE, Toshiba) and RHC. The HRCT images with 0.5 mm section thickness and 10 mm spacing were selected from all of the CT images in each patient. Using a semiautomatic image-processing program (Image J), CSA less than 5mm2 and 5-10mm2 , and total lung area were measured, and the percentage of the total CSA less than 5mm2 and 5-10mm2 for the lung area (%CSA<5 and %CSA5-10, respectively) were calculated. The correlations of %CSA with hemodynamics measured by RHC were evaluated.

Results:
Mean pulmonary arterial pressure (mPAP) and pulmonary vascular resistance (PVR) from RHC were 41±9 mmHg and 8.3±3.0 Wood Units, respectively. %CSA<5 and %CSA5-10 were 1.095±0.166% and 0.447±0.093 %, respectively. The correlation coefficients of %CSA<5 with mPAP and PVR were -0.45 (P<0.001) and -0.64 (P<0.001), respectively. The correlation coefficients of %CSA5-10 with mPAP and PVR were -0.20 (P=0.1) and -0.38 (P=0.002), respectively.

Conclusions:
This study suggests that %CSA<5 measured on HRCT is inversely correlated with PVR and can be used to evaluate hemodynamics in CTEPH subjects.
Abstract 13820: Diastolic Dysfunction is Not Associated With Increased Diastolic Calcium During Experimental Uremic Cardiomyopathy
Authors: Winterberg Pamela D , Jiang Rong , Wang Bo , Harbaran Sonal , Wagner Mary B
Published: July 2021

Introduction:
The underlying mechanisms contributing to uremic cardiomyopathy during chronic kidney disease (CKD) are poorly understood, limiting treatment options.

Hypothesis:
We aimed to determine if altered calcium (Ca2+) handling in cardiomyocytes contributes to diastolic dysfunction in a mouse model of CKD.

Methods:
CKD was induced in male 129X1/SvJ mice through five-sixths nephrectomy in a two-stage surgery. Age-matched mice served as controls. Transthoracic echocardiography and speckle-tracking based strain analysis (Vevo2100, VisualSonics, Toronto, Canada) were performed at 8 weeks post-CKD (n=7-8) to assess heart structure and function. Cardiomyocytes isolated from mice with or without CKD (n=3 mice per group, 10-12 cells/mouse) were loaded with Fura 2-AM, paced by field stimulation (1 Hz), and imaged with a dual-excitation fluorescence photomultiplier system (IonOptix Inc, Milton, MA) to measure Ca2+ transients and sarcomere length. Sarcoplasmic reticulum Ca2+ content was determined following rapid application of caffeine.[[Unable to Display Character:  ]]

Results:
CKD mice displayed left ventricular hypertrophy (LVAW;d 1.46 ± 0.134 vs 1.04 ± 0.129 mm; p<0.001) and decreased longitudinal strain (19 ± 4.1% vs 30 ± 2.3%; p<0.0001) compared to control mice. Resting sarcomere length was significantly shorter in cardiomyocytes isolated from CKD mice compared to normal mice (1.86 ± 0.054 vs 1.89 ± 0.047 nm; p = 0.016), but relaxation time was unchanged (0.21 ± 0.12 vs 0.21 ± 0.15 seconds, p=0.4). Unexpectedly, the baseline cytosolic Ca2+ content was lower in uremic myocytes (1.22 ± 0.353 vs 1.46 ± 0.252 AU, p=0.002). However, the Ca2+ transient amplitude (0.39 ± 0.177 vs 0.41 ± 0.167 AU, p=0.4) and sarcoplasmic reticulum Ca2+ content (1.15 ± 0.321 vs 1.24 ± 0.550 AU, p=0.4) were comparable between CKD and normal cardiomyocytes.[[Unable to Display Character:  ]]

Conclusions:
Mice with CKD have signs of left ventricular hypertrophy and diastolic dysfunction on echocardiography. Cardiomyocytes isolated from mice with CKD have shorter diastolic sarcomere length implying impaired relaxation, yet paradoxically have decreased diastolic calcium. Thus Ca2+ accumulation during diastole does not appear to contribute to impaired relaxation in this model.
Abstract 13874: Elevated [Na
+
]
i
in Cardiac Myocytes from Diabetic Rats Due to Enhanced Na
+
- Glucose Cotransport
Authors: Lambert Rebekah , Srodulski Sarah , Peng Xiaoli , Margulies Kenneth B , Despa Florin , Despa Sanda
Published: July 2021

Intracellular Na
+
concentration ([Na
+
]
i
) is a key regulator of cardiac Ca
2+
cycling, contractility and metabolism. [Na
+
]
i
is elevated in myocytes from failing hearts, leading to arrhythmias and oxidative stress. We hypothesized that myocyte [Na
+
]
i
is also increased in type-2 diabetes (T2D) due to enhanced activity of the Na
+
-glucose cotransporter (SGLT). To test this hypothesis, we used myocardial tissue from humans with T2D and an animal model of late-onset T2D (HIP rats). We found increased SGLT expression in failing hearts from patients with T2D compared to non-diabetic individuals (by 55±16%) and in HIP rat hearts (
vs
. age-matched wild-type, WT, littermates; by 59±17%). [Na
+
]
i
, measured with the fluorescent indicator SBFI, is increased in myocytes from diabetic HIP rats, both at rest (14.7±0.9 mM compared to 11.4±0.7 mM in WT) and during electrical stimulation at 2 Hz (17.3±0.8 mM
vs
. 15.0±0.7 mM in WT). However, the Na
+
/K
+
-pump function (measured as the rate of pump-mediated [Na
+
]
i
decline in intact myocytes) is not significantly altered in diabetic HIP rats. This result suggests that higher [Na
+
]
i
is due to an increased Na
+
entry in HIP rat myocytes. Indeed, Na
+
influx, assessed as the rate of [Na
+
]
i
rise upon Na
+
/K
+
-pump inhibition with 10 mM ouabain, was significantly larger in myocytes from diabetic HIP
vs
. WT rats (1.74±0.13 mM/min
vs
. 1.27±0.07 mM/min). SGLT inhibition with 250 μM phlorizin significantly reduced Na
+
influx in myocytes from diabetic HIP rats (to 1.08±0.20 mM/min), while it had no effect in the WT (1.14±0.21 mM/min). Phlorizin also significantly decreased glucose uptake in HIP rat myocytes (by 50±10 %) but not in WT, indicating an increased reliance on SGLT for glucose uptake in T2D hearts. In agreement with this result, the insulin-sensitive glucose uptake was greatly reduced in HIP rat myocytes
vs
. WT. These data suggest that SGLT is upregulated in diabetic hearts to compensate for reduced insulin-mediated glucose uptake. In summary, we found that [Na
+
]
i
is elevated in myocytes from diabetic HIP rats due to an increased Na
+
entry via the Na
+
-glucose cotransporter. Higher [Na
+
]
i
may contribute to arrhythmogenesis and oxidative stress in diabetic hearts.
Abstract 13888: Long-term Intermittent Fasting Treatment Improves Cardiac Function and Inotropic Reserve by Restoration of Cardiac Beta-adrenergic Signaling in an Experimental Model of Chronic Heart Failure
Authors: de Lucia Claudio , Gambino Giuseppina , Femminella Grazia D , Pagano Gennaro , Komici Klara , Petraglia Laura , Formisano Roberto , Elia Andrea , Cannavo Alessandro , Liccardo Daniela , D’Amico Maria L , Leosco Dario , Ferrara Nicola , Rengo Giuseppe
Published: July 2021

Introduction and Hypothesis:
Restricted diets are effective interventions to enhance cardiovascular function and metabolic profile and are known to improve life spam. IF (Intemittent fasting) dietary regimen has a cardioprotective effect in a rat model of myocardial infarction (MI) when diet is started before MI induction. In heart failure (HF), upregulation of G protein-coupled receptor kinase 2 (GRK2) contributes to dysfunctional beta-adrenergic receptor (βAR) signaling and to decrease cardiac inotropic reserve. Moreover, it has been shown that caloric restriction has positive effects on the development of left ventricular hypertrophy and improves ischemic tolerance. Hence, we will test whether a long-term restricted diet, started late after MI, is beneficial in HF and how it could affect cardiac adrenergic signaling.

Methods:
forty rats were randomly assigned to MI or sham operation. Four weeks later, a time point when post-ischemic HF was established, HF and sham rats were further randomized to a one year IF dietary restriction or ad libitum diet. Thus, our final animal population consisted in 4 groups: Sham normal diet, Sham IF diet, HF normal diet and HF IF diet.

Results:
One year of IF diet induced a robust decrease in body weight. In HF groups, restricted diet resulted in improved cardiac systolic function and reduced left ventricle end diastolic diameter compared to HF rats fed with normal diet, as measured by echocardiography at the end of the study period. No differences in cardiac function and dimension were observed between sham groups treated with different diets. Consistently, invasive hemodynamic showed that both LV contractility and relaxation in response to βAR stimulation were significantly increased in IF HF rats compared to HF normal diet animals. IF diet resulted in improved cardiac βAR density and adenylyl cyclase activity in HF rats when compared to HF rats treated with standard diet. Restoration of βAR signaling was associated to a dramatic reduction in cardiac GRK2 protein levels.

Conclusions:
We have demonstrated for the first time that IF, started when HF is already established, ameliorates cardiac function and inotropic reserve in an experimental model of HF. At the molecular level, IF diet significantly improves βAR signaling in HF.
Abstract 13907: Statin-Induced Myopathy is Mediated by Isoprenoid Depletion and is Independent of Serum Cholesterol Levels
Authors: Lawrence Jeffrey , Moyer Carolyn , Ameri Mehrdad , Dean Charley , Russell Betsy , Alexander Shlece , Cai Feng , Ason Brandon , Han HQ , Dent Ricardo , Xu Ren , Moffat Graeme
Published: July 2021

Statin use is associated with muscle-related side-effects (ie, MRSE) in ~10-20% patients and is the most common reason for discontinuation of treatment. Moreover, high-risk patients who have inadequately controlled LDL-C as a result of statin intolerance present a significant public health problem. A study to assess the role of cholesterol and/or isoprenoid intermediates of the cholesterol biosynthetic pathway in statin-induced myopathy was conducted in female rats treated with cerivastatin for 14 days. Cerivastatin treatment of rats fed standard diet induced myopathy in type II muscle fibers, but not in type I muscle fibers. Cholesterol supplementation in the diet of cerivastatin treated rats increased circulating cholesterol levels, but failed to prevent myopathy, suggesting that statin-mediated myopathy was independent of the cholesterol lowering effects of statins. Conversely, mevalonate supplementation of the diet abrogated cerivastatin induced myopathy evidenced by either microscopy or sensitive biomarkers of muscle injury including AST, CK, sTnI, FABP3, and urinary myoglobin. These data suggest that statin-induced myopathy involves a cholesterol biosynthetic intermediate upstream of cholesterol and downstream of mevalonate. Most animals with myopathy had increased expression of genes involved in stress (UCP1, NRF2), inflammation (Alox5ap), and autophagy/mitophagy (GABARAP), and decreases in genes involved in energy production (GATM, ATP5b, ATP5f1) or mitochondrial biogenesis (PGC-1α). These data suggest that sensitive muscle fibers may have deficits in mitochondrial biogenesis in the presence of increased autophagy/mitophagy that result in an energy crisis. In addition, many of these genes have also been reported to be changed in human statin myopathy, suggesting a similarity in mechanisms between rats and humans. In conclusion, cerivastatin-induced myopathy in rats involves a step upstream of cholesterol in isoprenoid intermediary metabolism that likely results in an energy crisis. These data suggest that lowering of cholesterol by mechanisms other than HMG CoA inhibition-associated isoprenoid intermediate depletion would not be expected to result in myopathy.
Abstract 13996: Does Copeptin Provide Additional Risk Stratification in Chest Pain Patients With a Mild Troponin Elevation?
Authors: Marston Nicholas , Shah Kevin , Mueller Christian , Xavier-Neath Sean , Christenson Robert , McCord Jim , Nowak Richard , Vilke Gary , Daniels Lori , Hollander Judd , Apple Fred , Cannon Chad , Nagurney John , Schreiber Donald , deFilippi Christopher , Hogan Chris , Diercks Deborah , Limkakeng Alexander , Anand Inder , Wu Alan , Clopton Paul , Jaffe Allan , Peacock Frank , Maisel Alan
Published: July 2021

Introduction:
Copeptin has demonstrated a role in early rule out for acute myocardial infarction in combination with a negative troponin. However, the value of copeptin in patients with a positive troponin is not established.

Hypothesis:
The addition of a baseline copeptin at chest pain presentation will improve risk stratification in patients with mild troponin elevation.

Methods:
The multi-center CHOPIN trial enrolled 2071 acute chest pain patients. All subjects with ST segment elevations were excluded from this analysis. Of the remaining patients, 124 had a mildly elevated troponin (defined by
<
2x URL) and were included in the study cohort. Baseline cTnI and copeptin levels were drawn on presentation, and another cTnI at 2 hours. Copeptin ≤14 pmol/l and troponin deltas ≤10% from baseline were considered positive. Two independent blinded cardiologists adjudicated AMI diagnosis. The value of a baseline copeptin in this cohort was assessed using AMI incidence, odds ratio, sensitivity, and negative predictive value (NPV).

Results:
Of the 124 patients in the study cohort, 73 (59%) had an elevated copeptin and an associated AMI incidence of 25% (18 AMIs). The remaining 51 patients (41%) were copeptin negative and diagnosed with 3 AMIs, an incidence of 5.9% (p = 0.006, figure 1). A positive copeptin increased the likelihood of AMI diagnosis with an odds ratio of 5.3 (95% CI: 1.5-19.2). The sensitivity and NPV for AMI were 86% and 94%, both of which were higher than the delta troponin (table 1). When an initial copeptin was combined with the delta troponin the sensitivity and NPV for AMI improved to 100%.

Conclusion:
Use of copeptin in chest pain patients with mild troponin elevation provides further risk stratification. The combination of copeptin with a delta troponin may aid in earlier AMI rule out in this subset of patients. This approach could be especially relevant as the increasing use of high sensitivity troponin assays leads to greater rates of mild troponin elevations.
Abstract 14007: Insulin Resistance Expressed by Homa2 Predicts Risk for Recurrent Coronary Events in Post Infarction Patients
Authors: Szepietowska Barbara , Kutyifa Valentina , McNitt Scott , Ryan Daniel H , Corsetti James P , Sparks Charles , Moss Arthur , Zareba Wojciech
Published: July 2021

We investigated risk for recurrent coronary events associated with insulin resistance in post infarction patients from the Thrombogenic Factors and Recurrent Coronary Events (THROMBO) study. The association between insulin resistance expressed by Homeostatic Model Assessment 2 for Insulin Resistance (HOMA2-IR) and risk for recurrent coronary events was investigated in a cohort of 1,035 patients evaluated 2 months after myocardial infarction with an average follow-up of 26 months. The end point for the study was recurrent coronary event defined as death, nonfatal myocardial infarction and unstable angina whichever occurred first. We used time dependent survival analysis and Cox proportional hazards regression method to determine the association between HOMA2 categorized as high >75th percentile and endpoints after adjustment for relevant clinical covariates. High HOMA2-IR defined as in fourth quartile(> 2.4) was associated with increased risk for recurrent coronary events (hazard ratio [HR] 1.54; 95% confidence interval [CI] 1.10-2.14; P <0.01) after adjustment for: age, gender, diabetes, prior myocardial infarction, pulmonary congestion on chest roentgenogram, ejection fraction< 0.3, PTCA and CABG. The interaction between HOMA2-IR and diabetes status regarding recurrent coronary events was not statistically significant. The highest risk of cardiac events was observed in non-obese patients (BMI<=30kg/m2) with high HOMA2-IR (hazard ratio [HR] 1.55; 95% confidence interval [CI] 1.05-2.23; P <0.05). Patients with High HOMA2-IR had higher plasma level of fibrinogen, PAI-1, von Willebrand factor, triglyceride and apolipoprotein B. Insulin resistance predicts recurrence of coronary events in post-infarction population. HOMA2-IR and BMI are useful parameters in clinical practice to stratify risk. Multilevel intervention to increase insulin sensitivity in people after myocardial infarction may modify clinical outcome.
Abstract 14013: Direct Comparison of Disease-Specific versus TALEN-Corrected iPSC-Derived Cardiomyocytes from Dilated Cardiomyopathy Patients
Authors: Ebert Antje , Riegler Johannes , Karakikes Ioannis , Termglinchan Vittavat , Mameen Mohammed , Wu Haodi , Shen Qi , Yeh Ting-Yuan , Gold Joseph , Wu Joseph C
Published: July 2021

Introduction:
Recent advances in regenerative medicine for cardiovascular disease (CVD) therapy focus on delivery of autologous induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) to recover heart function without immunosuppression. Here, we perform a direct comparison of iPSC-CMs from a healthy individual, a DCM patient, and following genome-correction with transcription activator-like effector nucleases (TALENs) both in vitro and in vivo.

Methods & Results:
We established patient-specific iPSC-CMs that recapitulate DCM disease-phenotypes in-vitro, including disrupted sarcomeres and impaired Ca2+ handling (p<0.05). We corrected the patient′s DCM-specific mutation R173W in the TNNT2 locus via TALEN-mediated footprint-free genome editing. Isogenic TALEN-corrected iPSC-CMs recovered functional properties comparable to healthy control. Ca2+ transient analysis revealed no significant difference between TALEN-corrected iPSC-CMs and healthy control regarding amplitude (ΔF/F0 4.71±0.55), decay (277.6 ±28.2 ms), and time to peak (240.5±22.5 ms). Confocal microscopy of TALEN-corrected iPSC-CMs confirmed sarcomeric structures as in healthy control (Fig 1). We transplanted (i) 1x10*7 DCM iPSC-CMs, (ii) 1x10*7 TALEN-corrected iPSC-CMs, and (iii) 1x10*7 healthy control iPSC-CMs into a subacute myocardial infarct (MI) rat model (n=15/group). Cell engraftment into the host myocardium was confirmed by immunohistochemistry. Comprehensive functional analysis via magnetic resonance imaging (MRI), echocardiography is underway.

Conclusions:
We generated TALEN-corrected iPSC-CMs from a DCM patient, which recover in vitro functional parameters of healthy control iPSC-CMs. We transplanted for the first time patient-specific and TALEN-corrected iPSC-CMs into a rodent MI model. This approach may represent a viable option for mechanistic analysis and regenerative medicine in CVD patients.
Figure 1:
Abstract 14015: Right Ventricular Remodeling is Additive to Global Longitudinal Strain in Predicting Long Term Survival in Patients with Pulmonary Hypertension
Authors: Goda Akiko , Ryo Keiko , Delgado Antonia M , Tayal Bhupendar , Simon Marc , Mathier Michael A , Gorcsan John
Published: July 2021

Introduction:
Right ventricular (RV) global longitudinal strain has been recently shown to be a marker of prognosis in patients with pulmonary hypertension (PH) but the additive role of RV remodeling on long term outcome is unclear.

Hypothesis:
Our aim was to test the hypothesis that RV remodeling measured by the RV area to left ventricular (LV) area ratio is additive to RV longitudinal strain in predicting prognosis in PH.

Methods:
We studied 120 PH patients, excluding patients with LVEF<50%, congenital heart disease, or coronary disease and 15 normal subjects. We measured the RV to LV area ratio in the 4 chamber view at end-diastole as an RV remodeling index and calculated the peak RV global longitudinal strain using 2D speckle tracking. Outcome events were predefined as death or lung transplantation over 8 years.

Results:
The RV-LV area ratio was larger in PH patients than in normal control subjects as expected (0.64±0.09 vs. 1.18±0.59, p<0.001). Using the RV global longitudinal strain cut off of -15% previously reported, it was significantly associated with death or lung transplant (AUC 0.629, p=0.0138). When the RV remodeling index RV-LV area ratio > 0.73 was combined with RV global longitudinal strain > -15%, patients had a significantly worse prognosis compared to RV strain value or RV remodeling index alone (p=0.0017; Hazard Ratio 2.29, 95% confidence interval of 1.34 to 3.9).

Conclusion:
RV remodeling by the echocardiographic RV-LV area ratio was additive to RV global longitudinal strain at predicting long term outcomes in PH patients and has promise for clinical applications.
Abstract 14036: An Algorithm to Guide Prognosis Based on the Combination of Peak VO
2
and V
E
-VCO
2
slope in Patients with HFrEF: The Henry Ford HospITal CardioPulmonary EXercise Testing (FIT-CPX) Project
Authors: Brawner Clinton A , Shafiq Ali , Aldred Heather A , Hassan Raakesh , Vasko Stephanie , Ehrman Jonathan K , Selektor Yelena , Tita Christina , Velez Mauricio , Williams Celeste T , Lanfear David E , Keteyian Steven J
Published: July 2021

Many studies have reported the prognostic significance of peak oxygen consumption (VO
2
) and V
E
-VCO
2
slope in patients with heart failure (HF). However, there are limited data stratifying risk based on a combination of these measures and how to best use them.

Purpose:
Describe 1 and 3-y event rates for the composite endpoint of mortality, left ventricular assist device (LVAD), or cardiac transplant (CT) based on the combined evaluation of peak VO
2
and V
E
-VCO
2
slope in patients with HF with reduced ejection fraction (≤ 40%; HFrEF).

Methods:
Patients (n= 1,116; 33% female; age= 54 ± 13 y) with a cardiopulmonary exercise test between 1997 and 2010 and confirmed HFrEF were identified. Endpoint data was obtained through 2011. Patients were grouped based on peak VO
2
(< 12, 12 to18, and > 18 mL/kg/min) and (V
E
-VCO
2
slope ≥ 34 or < 34). Cumulative events were identified from life tables. Cox regression with adjustment for age, gender, ejection fraction, and beta-blocker therapy was used to calculate the hazard ratio for V
E
-VCO
2
slope ≥ 34 within each peak VO
2
group.

Results:
The 1 and 3-y event rates are shown in the Table. Among patients with a peak VO
2
< 12, 1 and 3-y events were 23% and 44%, respectively. Within this group, V
E
-VCO
2
slope ≥ 34 represented more than twice the risk at both 1 y (HR 2.42, 95% CI 1.09, 5.38) and 3 y (HR 2.32, 95% CI 1.33, 4.05). Among patients with a peak VO
2
12 to 18, 1 and 3-y events were 14% and 30%, respectively. Within this group, a V
E
-VCO
2
slope ≥ 34 was associated with increased risk at both 1 y (HR 1.80, 95% CI 1.13, 2.87) and 3 y (HR 1.80, 95% CI 1.30, 2.50). Among patients with peak VO
2
> 18, 1 and 3-y events were 2% and 10%, respectively, and V
E
-VCO
2
slope was not statistically associated with increased risk.

Conclusion:
Among patients with a peak VO
2
≤ 18, V
E
-VCO
2
slope ≥ 34 further refines the risk for a composite endpoint of mortality, LVAD, or CT at both 1 and 3 y.
Abstract 14058: Intramural Ventricular Septal Defects After Repair of Conotruncal Anomalies Are Associated With Postoperative Morbidity
Authors: Patel Jyoti K , Ghosh Reena M , Glatz Andrew C , Ravishankar Chitra , Mascio Christopher E , Spray Thomas L , Cohen Meryl S
Published: July 2021

Introduction:
Intramural ventricular septal defects (VSDs) are tunnel-like communications between a great artery and right ventricular free wall trabeculations that can occur after repair of conotruncal anomalies. We sought to assess the prevalence of residual intramural VSDs and their impact on postoperative course.

Methods:
We reviewed all patients at our institution who underwent biventricular repair of a conotruncal anomaly involving VSD baffle to a great artery from the left ventricle from 1/1/11 to 6/30/13. Perioperative echocardiographic images were reviewed for residual intramural or non-intramural VSDs. The primary outcome was a composite of ECMO use, cardiac transplantation, or mortality. Secondary outcomes included hospital length of stay (LOS) and need for repeat catheter-based or surgical VSD closure.

Results:
Of 158 total subjects, 92 (58%) were male and diagnoses were tetralogy of Fallot (n=111), d-transposition of the great arteries (14), truncus arteriosus (13) and other (20). Median age at surgery was 51 days (IQR 6,107). Residual VSD was present in 111 subjects; 29 (26%) had an intramural defect, and 82 (74%) had non-intramural defects (71 peripatch defects, 6 small muscular defects, and 5 with both). Of the non-intramural VSDs, 79 (96%) were hemodynamically insignificant lesions (<2mm). Residual intramural VSDs were more likely than non-intramural VSDs to be greater than 2mm (48% vs 4%, p<0.001). There was no difference in weight, age at operation, or type of conotruncal anomaly based on type of residual VSD. The primary composite outcome was more prevalent in subjects with residual intramural defects compared to non-intramural defects (5/29 [17%] vs 4/82 [5%], p = 0.05). In addition, subjects with residual intramural VSDs had longer LOS (24 days [IQR 13,67] vs 11 days [IQR 5,23], p=0.002). The rates of repeat catheter-based or surgical closure were low overall but higher in subjects with intramural VSDs (8/29 [28%] vs 5/82 [6%], p=0.005).

Conclusions:
Intramural VSDs are common after repair of conotruncal anomalies and tend to be larger than non-intramural VSDs. Although most subjects still have favorable outcome, intramural VSDs are associated with greater postoperative morbidity, longer LOS, and need for reintervention.
Abstract 14641: Deficiency in the E3 Ubiquitin Ligase Parkin Exacerbates Alcoholic Cardiomyopathy: Role of Mitophagy
Authors: Xu Xihui , Ren Jun
Published: July 2021

Background:
Long-term heavy alcohol consumption has been shown to promote mitochondrial injury, unfavorable geometric and contractile changes in the heart. Parkin, a cytosolic E3 ubiquitin ligase encoded by PARK2 gene, plays an important role in the regulation of selective mitophagy. This study was designed to examine the role of Parkin in alcohol-induced myocardial injury (aka alcoholic cardiomyopathy) and the underlying mechanism with a focus on mitophagy.

Methods:
Adult male wild-type C57 and PARKIN2 knockout (Parkin-/-) mice were placed on alcohol (4%) or control diet for 4 weeks. Echocardiographic and cardiomyocyte mechanical properties were assessed. Mitochondrial morphology, function and mitophagy were examined using transmission electronic microscopy, Clark-type oxygen electrode, and Western blot, respectively.

Results:
Our results revealed that chronic alcohol consumption triggered unfavorable geometric and contractile changes [decreased fractional shortening (FS) and ejection fraction (EF), with enlarged left ventricular chamber; decreased peak shortening (PS) and velocity of shortening +dL/dt, increased time-to-90% relengthening TR90], the effects of which were exacerbated by Parkin deficiency. In addition, our data showed that chronic alcohol intake promoted myocardial mitochondrial swelling with cristae disarrangement, induced myocardial mitochondrial depolarization and respiration inhibition, which were exacerbated by Parkin knockout. Furthermore, chronic alcohol consumption promoted mitophagy activation, as evidenced by accumulation of Parkin and LC3BII in mitochondria and mitochondrial ubiquitination level in the heart, the effect of which was nullified by Parkin knockout.

Conclusion:
These data suggest that chronic alcohol consumption triggered mitophagy by stimulating Parkin translocation to the mitochondria, which may be an adaptive response in the heart. Our findings implicated the therapeutic potential of mitophagy as a target in the management of alcoholic cardiomyopathy.
Abstract 14647: Stress Echocardiography Predicts Heart Failure Progression and Alters Management Strategies in Patients with Hypertrophic Cardiomyopathy
Authors: Rowin Ethan J , Maron Barry J , Olivotto Iacopo , Casey Susan A , Arretini Anna , Tomberli Benedetta , Garberich Ross , Lesser John R , Maron Martin S
Published: July 2021

Background:
One-third of HCM patients without left ventricular outflow tract obstruction under resting conditions have the propensity to develop an outflow gradient with physiologic exercise. However, the natural history and management implications of exercise-induced (i.e., provocable) obstruction is unresolved.

Methods:
We prospectively studied 533 consecutive HCM patients without outflow obstruction at rest (<30mmHg) who underwent a symptom limiting stress (exercise) echocardiogram to assess development of outflow obstruction following physiologic provocation and followed for 6.5 ± 2.0 years. Of the 533 patients, obstruction ≥ 30 mmHg was present following exercise in 262 patients (49%; provocable obstruction), and was absent both at rest and with exercise in 271 (51%; nonobstructive).

Results:
Over the follow-up period, 43 out of 220 (20%) HCM patients with provocable obstruction and baseline NYHA class I/II symptoms developed progressive limiting heart failure symptoms to class III/IV, compared to 24 of 249 (10%) nonobstructive patients. Rate of heart failure progression was significantly greater in patients with provocable obstruction vs. nonobstructive patients (3.1%/year vs. 1.5%/year; RR=2.0, 95% CI of 1.3-3.2; p=0.003). However, the vast majority of patients with provocable obstruction who developed advanced heart failure symptoms achieved substantial improvement in symptoms to class I / II following relief of obstruction with invasive septal reduction therapy (n=30/32; 94%). In comparison, the majority of nonobstructive patients who developed advanced heart failure remained in class III/IV (16/24;67%), including 10 (42%) currently listed for heart transplant.

Conclusions:
Stress (exercise) echocardiogram identifies physiological provocable outflow tract obstruction in HCM, and is a predictor of future risk for progressive heart failure (3.1%/year), in patients who become candidates for invasive septal reduction therapy. Therefore, exercise echocardiography should be considered in all HCM patients without obstruction under resting conditions.
Abstract 14698: Self-Complementary AAV9 Provides No Advantage Over Single-Stranded AAV9 in shRNA Knockdown of Ubiquitously Expressed GFP in Heart or Liver after Systemic Delivery
Authors: Piras Bryan A , French Brent A
Published: July 2021

Introduction:
AAV9 is a potent gene delivery system that has been shown to preferentially transduce the heart over other tissues. We previously used AAV9 to achieve highly-efficient knockdown of GFP in heart but not liver of transgenic mice that express GFP from the human ubiquitin C promoter, and hypothesized that lack of liver knockdown was due to inefficient conversion of the single-stranded genomes into double-stranded DNA after AAV9-mediated delivery.

Methods:
An expression cassette containing a mouse U6 promoter driving shRNA against GFP was constructed using wild-type AAV2 ITRs (ssAAV) or a mutated 5’ ITR for packaging of self-complementary (scAAV) genomes into AAV9 capsids. ssAAV9 or scAAV9 vectors were administered to 10-day old mice via intraventricular chamber injection at a dose of 8х10^10 vg/g. Uninjected mice were used as controls. Cardiac and liver tissues were collected 7 weeks after injection for protein, RNA and genomic DNA. GFP expression was analyzed by qRT-PCR and western blotting.

Results:
Compared to controls (n=5) in cardiac tissue, the ssAAV group (n=8) demonstrated an 82% decrease in GAPDH-normalized GFP mRNA (p<0.0001) while the scAAV group (n=7) provided a 79% knockdown (p<0.0001) (Panel A). In liver, the ssAAV group showed a 35% reduction in GFP mRNA versus controls (p<0.01) and the scAAV group showed a 27% knockdown (p<0.05). Western blot analysis showed that GAPDH-normalized GFP expression was reduced by 51% in the ssAAV group (p<0.0001) and by 45% in the scAAV group (p<0.001) compared to controls in heart, but there was no reduction in GFP in the liver (Panel B, n=4 for all groups). Genomic DNA analysis demonstrated a 6-fold and 1.9-fold increase in viral genomes in liver vs. heart for the ssAAV group (p<0.0001) and scAAV group (p<0.05), respectively.

Conclusions:
AAV9 vectors expressing shRNA provide similar knockdown in cardiac and liver tissues regardless of whether they are packaged as single-stranded or self-complementary genomes.
Abstract 14741: Asian Dust Has an Impact on the Incidence of Acute Myocardial Infarction ~a Time-stratified Case-crossover Study
Authors: Kojima Sunao , Matsui Kunihiko , Ueda Kayo , Nitta Hiroshi , Ogawa Hisao
Published: July 2021

Introduction:
Air pollution, a mixture of gases and particles that come from manmade and natural sources, has been shown to increase hospi¬tal admission and mortality attributable to cardiovascular diseases. Asian dust (AD), a windblown sand dust originating from mineral soil in the desert, and PM2.5, which comes from cars and trucks, power plants, industrial boilers, other industrial sources and wildfires, are the standard for particulate matter representing air pollution.

Hypothesis:
We assessed the hypothesis that AD and PM2.5 were associated with the incidence of acute myocardial infarction (AMI) in Kumamoto prefecture.

Methods:
Kumamoto prefecture is located in southwestern Japan and has a stable population of 1.8 million. A total of 3,670 AMI patients in 2009-2012 in Kumamoto prefecture were entirely registered and 199 AMI patients whose onset date was not definite were excluded in the present study. A time-stratified case-crossover study was conducted to assess the association between particulate matters and AMI incidence. Within-subject comparisons were made between a case and control periods in this design. The study estimated the odds ratios and 95% confidence intervals associated with AMI incidence during the case day to a few previous days using conditional logistic regression.

Results:
The case-crossover analysis showed that PM2.5 was not significantly associated with the incidence of AMI (Figure upper). However, the results after adjustment for temperature and relative humidity revealed a close association between AD and the incidence of AMI (Figure lower). The strong association was observed on day 1. Adding co-pollutants including oxidants, nitrogen dioxide and sulfur dioxide in the basic model did not substantially alter the results.

Conclusions:
AD increases the incidence of AMI compared with PM2.5. Attention should be called to patients already at high risk for cardiovascular disease in case of an official announcement of AD warning.
Abstract 148: Zoniporide Combined with α-Methylnorepinephrine Appears Highly Effective for Resuscitation from Cardiac Arrest in a Rat Model of Ventricular Fibrillation
Authors: Lamoureux Lorissa , Whitehouse Herbert K , Radhakrishnan Jeejabai , Gazmuri Raúl J
Published: July 2021

Background:
We have reported in rat and swine models of cardiac arrest that sodium hydrogen exchanger isoform-1 (NHE-1) inhibition facilitates resuscitation, ameliorates myocardial dysfunction, and improves survival. Others have reported that α-methylnorepinephrine (α-MNE) - a selective α2-adrenoreceptor agonist - is superior to epinephrine given its lack of β-agonist effects. We examined in a rat model of VF and closed-chest resuscitation the effects of combining the NHE-1 inhibitor zoniporide (ZNP) with α-MNE.

Methods:
VF was electrically induced in 32 male retired breeder Sprague-Dawley rats and left untreated for 8 minutes after which resuscitation was attempted by an 8 minute interval of chest compression and delivery of electrical shocks. Rats were randomized 1:1:1:1 to receive a 3 mg/kg bolus of ZNP or 0.9% NaCl before starting chest compression and a 100 μg/kg bolus of α-MNE or its vehicle at minute 2 of chest-compressions establishing 4 groups of 8 rats each. Successfully resuscitated rats were monitored for 240 minutes.

Results:
The number of rats that had return of spontaneous circulation and then survived 240 min were: α-MNE(-)/ZNP(-) 4 and 2; α-MNE(-)/ZNP(+) 5 and 5; α-MNE(+)/ZNP(-) 2 and 1; and α-MNE(+)/ZNP(+) 7 and 7 yielding a statistically significant effect on overall survival times corresponding to 105 ± 114, 150 ± 124, 58 ± 108, and 210 ± 85 min, respectively (p < 0.045). Post-resuscitation lactate levels were attenuated in all treatment groups with the greatest effect by the α-MNE(+)/ZNP(+) combination without major differences in hemodynamic function (Table).

Conclusion:
We confirm a beneficial effect resulting from the combination of ZNP (given to attenuate myocardial reperfusion injury) and α-MNE (given to augment peripheral vascular resistance during chest compression without the detrimental actions of epinephrine). The proposed combination may prove to be a highly effective novel strategy for resuscitation from cardiac arrest.
Abstract 14864: Risk Markers for Coronary Plaque Progression and Destabilization Beyond LDL-Cholesterol in Patients with Acute Coronary Syndrome
Authors: Komatsu Takaaki , Taguchi Isao , Nakahara Shiro , Kobayashi Sayuki , Sakai Yoshihiko , Kageyama Michiya , Sakuma Masashi , Toyoda Shigeru , Inoue Teruo
Published: July 2021

Introduction and Purpose:
In acute coronary syndrome (ACS) patients, it is important to stabilize pan-coronary vulnerable plaques as early as possible in ACS patients, in addition to revascularization of the culprit lesion by percutaneous coronary intervention (PCI). Although statins are effective for regression and stabilization of vulnerable plaques in ACS patients, several patients experience plaque progression and/or destabilization despite aggressive control of low-density lipoprotein (LDL)-cholesterol by statins. Therefore, we should do further efforts to research new risk markers beyond LDL-cholesterol and to intervene in them.

Methods and Results:
In 57 patients with ACS, who underwent emergent percutaneous coronary intervention (PCI), we performed a virtual histology (VH)-IVUS for a non-culprit lesion, at baseline and at the follow-up time (8-10 months), to assess plaque regression/progression and plaque stabilization/destabilization. At the follow-up time, we measured specific biomarkers. All of the patients received aggressive treatment with statins. Change in the total plaque volume by the gray scale IVUS was correlated positively with derivatives of reactive oxygen metabolites (d-ROMs) level (0.32, P<0.05). Change in the volume of destabilized components, i.e., necrotic core plus fibro-fatty compositions, was correlated negatively with the eicosapentaenoic acid to arachidonic acid (EPA/AA) ratio (R=-0.39, P<0.01) and positively with the levels of interleukin (IL)-8 (R=0.31, P<0.01) and d-ROMs (0.32, P<0.01). Multivariate logistic regression analysis showed that only the d-ROMs level was an independent discriminative biomarker for plaque progression (hazard ratio; 1.018, 95% confidence interval; 1.005-1.032, P<0.01) and plaque destabilization (hazard ratio; 1.022, 95% confidence interval; 1.005-1.038, P<0.01).

Conclusions:
The d-ROMs would be a powerful biomarker to predict plaque progression and destabilization as the residual risk marker beyond LDL-cholesterol. Intervention to oxidative stress state would be the most important residual target beyond LDL cholesterol lowering for plaque regression and stabilization.