bluuebunny/crossref_metadata_2025 huggingface dataset:
The Effects of Chronic Dialysis on Physical Status, Quality of Life, and Arterial Stiffness: A Longitudinal Study in Prevalent Dialysis Patients
Authors: Gadaen Rens J.R. , Kooman Jeroen P. , Cornelis Tom , van der Sande Frank M. , Winkens Bjorn J. , Broers Natascha J.H.
Published: October 2020

Introduction: It is widely known that dialysis patients have significantly impaired functional outcomes and arterial stiffness, but still few studies have investigated the effects of dialysis longitudinally by a multidimensional approach. We aimed to assess longitudinal patterns of physical activity (PA), physical functioning (PF), health-related quality of life (HrQoL), body composition (BC), and arterial stiffness in prevalent dialysis patients. Materials and Methods: Thirty-nine prevalent dialysis patients (23 conventional hemodialysis [CHD] and 16 peritoneal dialysis) with a mean vintage of 25.7 (±22.1) months were included in this observational prospective study with a 2-year follow-up, and at baseline 20 healthy controls were included. Measurements were performed every 6 months. HrQoL was assessed using the Short Form-36 (SF-36) questionnaire. PA was assessed using the SenseWear™ Pro3 accelerometer. PF was assessed by walking speed, the PF subscale of the SF-36, and handgrip strength (HGS). BC was assessed using the Body Composition Monitor® and arterial stiffness by measuring carotid-femoral pulse wave velocity (PWV). The longitudinal trend was assessed using linear mixed models, correcting for sex, age, and dialysis vintage. For PWV, the trend was additionally corrected for diabetes and systolic blood pressure. Results: After correction, no statistically significant changes over time were observed for the parameters of PA, PF, HrQoL, and BC. In the combined group and in the group of CHD patients only, a significant change was observed for PWV (overall trend: p = 0.007 and p = 0.008, respectively). A statistically significant difference at baseline was observed between dialysis patients and healthy controls in all parameters, except for HGS and PWV. Discussion/Conclusion: We observed no statistically significant changes in functional outcomes during a 2-year follow-up period, but a significant increase was observed for arterial stiffness. These results might suggest that after a certain period in time, a relatively stable course is present in functional outcomes, but an ongoing deterioration in arterial stiffness occurs, which might increase the risk of cardiovascular disease and all-cause mortality in these patients.
Selective Immunoglobulin A Deficiency, Helicobacter pylori Infection, and Strongyloidiasis in a Patient with Adenocarcinoma of the Stomach
Authors: Dakkak Bassel Mazen , Imam Yahia , Ahmed Khalid , Saleh Ahmed Osman , Soliman Dina , Alanzi Meshaal , Yassin Mohamed A. , Mohamed Shehab
Published: October 2020

Selective immunoglobulin A (IgA) deficiency is one of many congenital immunodeficiencies. It is associated with several medical condition. It has been shown to be associated with some types of malignancies, some autoimmune disorders, and even with some infections. Here we report a young male with selective IgA deficiency who also tested positive for Helicobacter pylori and strongyloidiasis at the time when he was diagnosed with stomach adenocarcinoma. The presence of IgA deficiency with multiple etiological possibilities such as infections and cancer makes this case unusual.
Концепт „лишние люди“ и его динамика в русской драматургии XIX века
Authors: Dimitrov Lyudmil
Published: October 2020

Понятие «лишние люди» возникло в русской литературе в 1950-х годах и постепенно было перенесено во внелитературную среду, став идеологемой. В статье предпринята попытка исследовать этот феномен в пространстве русской драматургии, начиная с Грибоедова вплоть до драматургов конца XIX - начала XX века Льва Толстого, Антона Чехова и Максима Горького, которые являются представителями трех разных поколений, а также трех эстетических направлений социокультурной мысли в России, оказывая огромное влияние на развитие европейского театра в XX и XXI веках.
A era do estranhamento: o Formalismo Russo e o conhecimento humanístico moderno
Authors: Vaz Valteir
Published: October 2020

Esta resenha apresenta ao leitor brasileiro o livro A era do estranhamento: o Formalismo Russo e o conhecimento humanístico moderno, publicado em 2017. O livro é resultado do evento internacional ocorrido em Moscou em 2013, que estabeleceu e celebrou o centenário do Formalismo Russo. A obra apresenta contribuições de pesquisadores do mundo todo, revisa e expande, consideravelmente, os limites de alcance de uma das mais importantes escolas de crítica literária do século XX.
Comparison Between Optical Frequency Domain Imaging and Intravascular Ultrasound for Percutaneous Coronary Intervention Guidance in Biolimus A9-Eluting Stent Implantation
Authors: Muramatsu Takashi , Ozaki Yukio , Nanasato Mamoru , Ishikawa Masato , Nagasaka Ryo , Ohota Masaya , Hashimoto Yosuke , Yoshiki Yu , Takatsu Hidemaro , Ito Katsuyoshi , Kamiya Hiroki , Yoshida Yukihiko , Murohara Toyoaki , Izawa Hideo ,   on behalf of the MISTIC-1 Investigators
Published: October 2020

Background:

Given the characteristic differences between intravascular ultrasound (IVUS) and optical frequency domain imaging (OFDI), their approach to therapeutic guidance during percutaneous coronary interventions (PCIs) and arterial healing response after stenting may also vary.

Methods:

MISTIC-1 (The Multimodality Imaging Study in Cardiology cohort 1) is a multicenter, randomized-controlled, noninferiority trial that compared imaging end points between OFDI- and IVUS-guided PCI. Patients with stable coronary artery disease were randomly assigned to either OFDI- or IVUS-guided PCI using a Biolimus A9-eluting stent according to a prespecified protocol for imaging guidance. Stent sizing was based on external elastic lamina in IVUS-guided PCI while lumen up-size in OFDI-guided PCI. Postprocedural OFDI was investigated regardless of randomization, while operators in IVUS-guided PCI arm were blinded to the images. The primary end point was in-segment minimum lumen area assessed using OFDI at 8 months, while the secondary end point was a composite of cardiovascular mortality, target-vessel myocardial infarction, or target-lesion revascularization (device-oriented composite end point). Patients were followed up to 3 years after the index procedure.

Results:

A total of 109 patients (mean age 70 years, male 78%) with 126 lesions were enrolled. Postprocedural minimum stent area was 6.31±1.89 and 6.72±2.08 mm
2
in OFDI and IVUS group, respectively (
P
=0.26). At the 8-month follow-up, in-segment minimum lumen area was 4.56±1.94 and 4.13±1.86 mm
2
in OFDI and IVUS group, respectively (
P
non-inferiority
<0.001). Both groups had comparable neointimal healing score (median 0.16 [interquartile range, 0.00–3.14] versus 0.90 [0.00–3.30], respectively;
P
=0.43). The incidence rate of device-oriented composite end point at 3 years was 7.4% and 7.3% in OFDI and IVUS group, respectively (hazard ratio, 1.05 [95% CI, 0.26–4.18];
P
=0.95).

Conclusions:

OFDI-guided PCI was not inferior to IVUS-guided PCI in terms of in-segment minimum lumen area at 8 months. Although a small sample size was acknowledged, OFDI could be an alternative to IVUS when considering intracoronary imaging-guided PCI in selected populations with coronary artery diseases.

Registration:

URL:
https://www.clinicaltrials.gov
. Unique identifier: NCT03292081.
Shortening Ambulance Response Time Increases Survival in Out‐of‐Hospital Cardiac Arrest
Authors: Holmén Johan , Herlitz Johan , Ricksten Sven‐Erik , Strömsöe Anneli , Hagberg Eva , Axelsson Christer , Rawshani Araz
Published: October 2020

Background

The ambulance response time in out‐of‐hospital cardiac arrest (OHCA) has doubled over the past 30 years in Sweden. At the same time, the chances of surviving an OHCA have increased substantially. A correct understanding of the effect of ambulance response time on the outcome after OHCA is fundamental for further advancement in cardiac arrest care.

Methods and Results

We used data from the SRCR (Swedish Registry of Cardiopulmonary Resuscitation) to determine the effect of ambulance response time on 30‐day survival after OHCA. We included 20 420 cases of OHCA occurring in Sweden between 2008 and 2017. Survival to 30 days was our primary outcome. Stratification and multiple logistic regression were used to control for confounding variables. In a model adjusted for age, sex, calendar year, and place of collapse, survival to 30 days is presented for 4 different groups of emergency medical services (EMS)‐crew response time: 0 to 6 minutes, 7 to 9 minutes, 10 to 15 minutes, and >15 minutes. Survival to 30 days after a witnessed OHCA decreased as ambulance response time increased. For EMS response times of >10 minutes, the overall survival among those receiving cardiopulmonary resuscitation before EMS arrival was slightly higher than survival for the sub‐group of patients treated with compressions‐only cardiopulmonary resuscitation.

Conclusions

Survival to 30 days after a witnessed OHCA decreases as ambulance response times increase. This correlation was seen independently of initial rhythm and whether cardiopulmonary resuscitation was performed before EMS‐crew arrival. Shortening EMS response times is likely to be a fast and effective way of increasing survival in OHCA.
Abstract 101: Novel Inhibitors for Temporal Modulation of T-lymphocytes During Chronic Heart Failure
Authors: Bansal Shyam S , Rosenzweig Rachel , NEHRA SARITA , Bermeo Blanco Oscar , Stratton Matthew , Gumina Richard J , Bennett Chad , Coss Christopher
Published: October 2020

CD4+ T-cells mediate wound-healing post-myocardial infarction (MI) but exacerbate left-ventricular (LV) remodeling during chronic heart failure (HF). Mechanisms that lead to this transition are unknown. Therefore, we hypothesized that as opposed to MI, T-cells activate specific pathological signals to promote LV-remodeling during chronic HF. To identify such signals, we conducted limited cell RNA-sequencing of CD4+ T-cells sorted from the failing hearts (8 weeks post-MI) of male mice and, surprisingly, found activation of estrogen receptor (ER)-α signaling. As ERα effects are antagonized by ERβ, we tested a novel ERβ agonist (OSU-ERB-012) to modulate T-cell activity and LV remodeling.
In-vitro
assays showed that OSU-ERB-012 dose dependently inhibit activation and proliferation of T-cells isolated from male mice (IC
50
3.4 μM).
In-vivo
assays (60 mg/kg/day; oral) showed no overt toxicity and a significant reduction in circulating T cells without affecting neutrophils, monocytes, or B-cells indicating specificity against T-cells. Furthermore, this effect was specific to TCR-mediated T-cell activation and the drug did not affect T-cells stimulated with PMA/Ionomycin suggesting preferential inhibition of antigenically-activated T-cells. To test therapeutic efficacy, male 10-12 week old mice underwent coronary ligation or sham operation and at 4 weeks post-MI randomized according to their cardiac function to receive either the vehicle or OSU-ERB-012 (60 mg/kg/day, oral) for the next 4 weeks. Consistently, at 8 weeks we observed a significant reduction in T-cells in the circulation and the spleens of drug-treated mice when compared with the vehicle treatment. As expected, vehicle treated HF mice showed progressive LV dilatation with significantly increased end-diastolic and end-systolic volumes (EDV and ESV, respectively) from 4-8 weeks post-MI. Importantly, treatment with OSU-ERB-012 significantly inhibited these changes and blunted LV remodeling from 4-8 weeks post-MI. Significant reduction in tibia-normalized heart weights supported these results. Our studies indicate that OSU-ERB-012 is a novel orally-active drug molecule that selectively inhibits T-cell activation and blunts pathological LV-remodeling during chronic HF.
Abstract 102: Sympathetic Innervation Negatively Regulates Postnatal Cardiomyocyte Proliferation Through Circadian Genes
Authors: Tampakakis Emmanouil , Murphy Sean , Gangrade Harshi , Kowalski William , Mukoyama Yosuke , Kwon Chulan
Published: October 2020

Sympathetic neurons (SNs) regulate heart rate, conduction velocity, contractility and relaxation of the myocardium. Disruption of SNs in adult cardiac disease, leads to arrhythmias, myocardial dysfunction and sudden cardiac death. However, the role of SNs during cardiac development and whether disruption of cardiac SNs at embryonic stages is associated with disease progression in adult life is unknown. Moreover, controversy exists regarding the effect of SNs on neonatal myocardial regeneration after injury. As the heart is innervated by SNs from mid-gestation and the innervation continues through the neonatal stage, we generated genetically modified mice with profoundly reduced cardiac-specific sympathetic innervation starting at embryonic stages. Inhibition of sympathetic innervation resulted in larger heart size with increased number of myocytes that were smaller and more mononuclear. Analysis also confirmed increased cardiomyocyte proliferation. Interestingly, transcriptomic analysis of P7 and P14 hearts verified dysregulated cell cycle, calcium homeostasis and circadian genes. Remarkably, this led to persistently increased heart size and reduced function at adult stages. To investigate the mechanism whereby SNs, affect cardiomyocyte proliferation, we first co-cultured human stem cell-derived or mouse cardiomyocytes with ganglionic SNs and confirmed similar gene expression patterns. Moreover, alpha1 and beta2-specific adrenergic receptor agonists and norepinephrine, reduced cell cycle genes and myocyte proliferation and upregulated circadian genes such as Period1 and Period2. As circadian genes have been linked with regulation of cell cycle and apoptosis, we analyzed neonatal hearts from Period1/Period2 DKO mice and curiously discovered increased heart size, cardiomyocyte proliferation and cell cycle genes, and suppression of the Wee1 kinase an inhibitor of cell mitosis. More importantly, norepinephrine did not alter cardiomyocyte proliferation and cell cycle genes in Period1/Period2 DKO myocytes. To our knowledge this is the first study to provide direct evidence of the effect of SNs in the regulation of circadian genes in the heart and their role on neonatal cardiomyocyte proliferation.
Abstract 103: TBX20 Activates Cardiac Maturation Gene Programs Promoting Direct Human Cardiac Reprogramming
Authors: Zhou Yang , Tang Yawen , Fast Vladimir G , Zhao Lianzhong , Lu Rui , Zhang Jianyi
Published: October 2020

Direct cardiac reprogramming to generate induced cardiomyocyte like cells (iCMs) from fibroblasts has emerged as a promising therapeutic strategy for the treatment of heart failure, which is still the leading cause of mortality and morbidity in developed countries. While much is known regarding iCMs generated from mouse cells, the adaptation of direct cardiac reprogramming to human cells is hurdled with low efficiency and poor quality because of intrinsic differences between species. Recently, our single cell transcriptomic analysis during human direct cardiac reprogramming demonstrated the immature features of human iCMs (hiCMs) compared to endogenous cardiomyocytes (CMs) at the transcriptome level, suggesting that key factors required for CM maturation remain unidentified. We then utilized computational analyses of genomic and epigenomic data to identify the regulators underexpressed in hiCMs comparing to CMs derived from human heart tissue or stem cell differentiation. Among them, the top hit is T-box family transcription factor TBX20, whose role in direct cardiac reprogramming remains to be explored. Supplementing TBX20 in reprogramming cocktails, we found significantly increased sarcomeric protein expression, like αMHC and αActinin in hiCMs and the formation of well-organized myofibrils. TBX20’s effect has also been supported by transcriptomic analysis, where we found TBX20 activates a large number of underexpressed genes associated with sarcomere structure and ion channels, such as
MYH7
,
MYL2
,
MYBPC3,
SCN5A
and
KCNQ1
, suggesting that TBX20 likely enhances maturation and contractility of hiCMs. We also found that TBX20 cannot replace any of the reprogramming factors, suggesting that its potential role is complimentary to and interacting with the current cocktail. Moreover, known TBX20 targets showed inconsistent alteration in TBX20-transduced hiCMs, suggesting its novel regulation in a direct reprogramming setting. These findings assign a novel reprogramming factor TBX20 playing an essential role in generating mature hiCMs by establishing gene programs associated with cardiac muscle contractility.
Abstract 203: Appropriate Utilization of the HEART Score Saves Money: A Community Hospital Perspective
Authors: Tran Ly , Ervin Charlie , Mickalis Morgan , Weigle Rachael , Sampath Rahul , Isserman Steven
Published: October 2020

HEART Score (HS) is a risk stratification tool used in predicting major adverse cardiac events (MACE) in patients presenting with undifferentiated chest pain. It is calculated by attributing zero, one, or two points in the following domains: history, EKG, age, risk factors, and troponin with a composite score of zero to ten. Low-risk patients with HS ≤3 are considered safe for outpatient evaluation. Prior studies show appropriate utilization of the HS reduces hospitalizations and unnecessary cardiac testing as well as healthcare cost. We conducted a retrospective chart review of young adults <45 years of age, admitted from the emergency department (ED) with undifferentiated chest pain between August 2018 and September 2019. HS was calculated for each adult, and when available, compared to scores calculated by the ED. Patients were reviewed for 30 day MACE, HS accuracy, diagnostic testing, and hospitalization cost. Of our 150 admitted patients, 91 (60.7%) had a HS ≤3 representing a low-risk cohort. HS was calculated by the ED in 36 patients (24%), 11 of those 36 were low-risk. 23 out of 36 patients had scores different from our calculation. No MACE occurred in the low-risk group. A single angioplasty was performed in a patient with a HS of 5. Median charge on each admitted patient with HS ≤3 is $19,000, translating to 1.73 million dollars.In a rural community hospital, outpatient management of young adults age ≤45 with a HS≤3 appears safe and cost-effective. In our institution, the HS was underused with score variability among providers. We feel there is an opportunity to improve care and save money through proper implementation of the HS. This is achievable through education, standard protocols, and better communication among ED, in-patient, and ambulatory services.
Abstract 204: Modification of AIFm2 By 4HNE Leads to Retrograde Stress Signaling During Ischemia- Reperfusion Injury in Heart
Authors: Sharma Sudha , Bhattarai Susmita , Subedi Utsab , Acosta Christina , Ara Hosne , Bhuiyan Shenuarin , Panchatcharam Manikandan , Miriyala Sumitra
Published: October 2020

Myocardial infarction is a leading cause of death worldwide and occurs due to blockage in blood supply to the heart. Re-establishment of blood flow after a brief period of ischemia leads to paradoxical exacerbation of the cardiomyocyte and its death. This phenomenon is known as ischemia-reperfusion injury. Major evidence in the pathogenesis of ischemia-reperfusion injury is due to oxidative stress, which is an imbalance between reactive oxygen species (ROS) and antioxidants. Membrane consisting of polyunsaturated fatty acid is attacked by ROS leading to lipid peroxidation and the generation of one of the toxic aldehydes 4-hydroxynonenal (4-HNE). Evidence suggests that 4-HNE increases during an ischemia-reperfusion injury in the heart. Apoptosis-inducing factor, mitochondrion-associated 2 (AIFM2) is a mitochondrial located oxidoreductase that participates in caspase-independent apoptosis. In this study, we sought to identify the role of 4-HNE in regulating AIFm2 translocation and cardiomyocyte death during an ischemia-reperfusion injury in the heart. Following ischemia both RNA and protein expression of AIFm2 significantly increased in the ischemic heart compared to sham. Also, 4-HNE adducted AIFm2 translocated from mitochondria to the nucleus shown by western blot analysis in ischemic heart. The mass spectrometric analysis was done to see the modification site on AIFm2 by 4-HNE and revealed that His 174 and Cys 187 are two sites on AIFm2 where 4-HNE adduction occurred. To identify the modification site responsible for AIFm2 translocation we performed site-direct mutagenesis in H9C2 cardiomyocyte, where Histidine 174 was replaced by arginine and Cys 187 was replaced by threonine. When the ischemia-reperfusion injury was induced, only Histidine 174 mutant failed to translocate to the nucleus indicating His 174 modification by 4-HNE was responsible for AIFm2 translocation. To further support the transport mechanism, protein expression of Importin; an adaptor protein responsible for the transfer of proteins to the nucleus was increased in the ischemic heart compared to sham. Collectively, those results for the first time identify the unique role of 4-HNE modification on AIFm2 protein during an ischemia-reperfusion injury in the heart.
Abstract 206: Serum Sitosterol Levels and ABCG5 and ABCG8 Genetic Mutations Among Japanese Dyslipidemic Subjects
Authors: Tada Hayato , Kojima Nobuko , Usui Soichiro , Kawashiri Masa-aki , Takamura Masayuki
Published: October 2020

Background:
Currently, serum concentrations of sitosterol above 10 μg/ml are considered to be one of the major diagnostic criteria of sitosterolemia. However, few data exist regarding the validation of this cutoff value from the viewpoint of predicting the presence of sitosterolemia with double pathogenic mutations in
ABCG5
or
ABCG8
genes.

Methods:
We retrospectively investigated consecutive 206 Japanese dyslipidemic subjects (mean age = 47 yr, male = 101) with the assessments of serum sitosterol level and the presence of
ABCG5
or
ABCG8
genetic mutations in our institute since 2009 to 2018. Exome lesions of those 2 genes were sequenced. We divided the subjects into 3 groups based on the number of pathogenic mutations in
ABCG5
or
ABCG8
genes. We compared serum lipids and serum sitosterol among those groups, and tried to validate the cutoff value discriminating patients of sitosterolemia with double mutations from others.

Results:
We identified 8 individuals with sitosterolemia with double mutations, 26 individuals with a single mutation, and 172 individuals without any mutations. Serum sitosterol level of patients with sitosterolemia with double mutations exhibited significantly higher than those of any other groups (52.0 vs. 7.9 μg/ml, p= 2.22х10
-6
, 52.0 vs. 2.8 μg/ml, p=2.275х10
-8
). Under these conditions, a cutoff value of sitosterol 10 μg/ml could discriminate the patients with sitosterolemia with double mutations in
ABCG5
or
ABCG8
gene from no mutation carrier perfectly, although 6 heterozygous mutation carries exhibited sitosterol level greater than 10 μg/ml.

Conclusion:
A cutoff value of sitosterol 10 μg/ml was well validated as an essential part of diagnostic criterion for sitosterolemia.
Abstract 216: Establishing an Integrated Platform to Validate Genetic Targets Associated With Atrial Remodeling
Authors: Ang Yen-Sin , Yu John , Li Ning , Ndungu Joan , Rajamani Sridharan
Published: October 2020

Atrial fibrillation (AF)–the most common form of arrhythmia in the general population– is a grievous illness and a public health concern of global and epidemic scale. Current pharmacological therapies targeting ion channels or β-receptors are only partially effective and often fail to reduce AF burdens or mortality. We hypothesized that structural atrial remodeling, in addition to electrical remodeling, are key drivers of AF initiation and progression. In order to discover unrecognized drivers of AF mechanisms and reduce attrition rates in drug development, we employed Genome-Wide Association Studies (GWAS) to identify novel targets with human genetic validation. Meta-analyses of multi-ethnic patient cohorts reported 134 genetic loci, and therefore hundreds of coding variants, that are associated with AF. Atrial-specific cardiomyocytes (aCMs) differentiated from induced pluripotent stem cells (iPSCs) offer a human-relevant model system to study atrial remodeling. We combined aCMs with phenotyping platforms like Multi-Electrode Array (MEA) and Traction Force Microscopy (TFM), to evaluate whether unknown genes play a role in either cardiac conductance and/or contraction. We first devised a multi-factorial computational ranking algorithm based on WASPAS to select 50 out of 187 protein-coding targets. Next, we performed a focused loss-of-function phenotypic screen in aCMs to determine their requirement for atrial myocyte function. This resulted in the prioritization of six high value targets with diverse roles in Rho signaling, cell migration, ER stress response and integrin signaling. We also developed efficient CRISPR/Cas9 gene editing of GWAS variants as well as gain-of-function methods in aCMs. The completion of this project will establish an integrated platform for the functional validation of human genetic targets emerging from population studies associated with atrial remodeling.
Abstract 220: Emerging Roles of Altered NAD Metabolism in Diabetic Cardiomyopathy
Authors: Lee Chi Fung
Published: October 2020

Diabetes is linked to altered NAD metabolism whose causal roles in cardiac dysfunction are poorly understood. Synthesis, consumption and redox balance of NAD dictate NAD metabolism and its homeostasis, and we here explored their roles in diabetic cardiomyopathy. 16 week of Type 1 diabetes (T1D) to C57BL6N mice was associated with lowered cardiac NAD/NADH, mild systolic and more severe diastolic dysfunction. We next used mouse models with altered NAD metabolism to determine how NAD redox balance impacts diabetic cardiomyopathy. Using cardiac-specific Ndufs4-KO mice (cKO) as a model of latent decrease in cardiac NAD/NADH, we observed accelerated systolic and diastolic dysfunction (lowered fractional shortening, E’/A’ ratio, and increased e/E’ ratio) in male cKO mice stressed with chronic T1D. Cardiac hypertrophy (heart weight/tibia length), insulin depletion and hyperglycemia levels were similar in these mice. Serum metabolomic analyses (~240 metabolites) also showed unchanged aqueous and lipid metabolite levels, suggesting that the diabetic stresses on these hearts were similar. The accelerated dysfunction of T1D cKO hearts was also observed in another female cohort. Importantly, elevation of cardiac NAD levels to attenuate NAD redox imbalance mitigated the accelerated dysfunction of T1D cKO hearts. In another cohort, control and cKO mice were stressed by 16-week high fat diet feeding. Accelerated systolic and diastolic dysfunction, and increased hypertrophy were observed in T2D cKO mice with similar glycemic levels to control mice. The data suggested that NAD redox imbalance is a positive regulator of cardiac dysfunction induced by T1D or T2D. However, NAD-dependent pathogenic mechanism induced by T1D or T2D (e.g. hypertrophy) can be different, and is under further investigation. Tissue fibrosis and mRNA levels of pro-fibrotic genes, including Adamts proteinases, integrins, laminins, matrix metalloproteinases and collagens were unchanged in T1D cKO hearts. Therefore, the accelerated dysfunction of T1D cKO hearts is due to cardiomyocyte dysfunction. We examined how NAD metabolism, other than the redox balance, may alter cardiomyocyte function. Levels of metabolite and mRNA regulating NAD synthesis and consumption pathways were measured. Of 32 transcripts assayed,
Nmrk2
levels were uniquely up-regulated in T1D cKO hearts, and lowered by elevation of cardiac NAD levels. NMRK product levels were concomitantly decreased in T1D cKO hearts. NAD-dependent global acetylation and inhibitory superoxide dismutase 2 acetylation were increased in T1D cKO hearts. Protein oxidation levels were concomitantly raised. Hyperacetylation in proteins like CaM kinase was associated with increased phosphorylation in troponin I, not in MyBPC or PLN in T1D cKO hearts. These data support the emerging, multifaceted roles of altered NAD metabolism in the progression of diabetic cardiomyopathy.
Abstract 222: The Estrogen-related Receptor (ERR) Drives Cardiac Myocyte Maturation in Cooperation With GATA4
Authors: Sakamoto Tomoya , Wan Shibiao , Batmanov Kirill , Kelly Daniel P
Published: October 2020

During transition from fetal to adult developmental stages, the heart undergoes robust mitochondrial biogenesis in parallel with growth and structural maturation. Recently we identified a role for ERRα/γ in this broad program of postnatal cardiac maturation. We sought to delineate the mechanisms whereby ERRs coordinate transcriptional regulation of both metabolic and structural genes in the maturation of human induced pluripotent stem cell-derived cardiac myocytes (hiPSC-CMs). CRISPR-based gene deletion studies demonstrated that ERRα/γ is necessary for activation of genes involved in mitochondrial and structural maturation during hiPSC-CM differentiation including mitochondrial function, ion transport, Ca
2+
handling, and the adult sarcomere. Whole-genome RNA-sequencing and ChIP-sequencing (ChIP-seq) studies indicated that ERRγ directly regulates both metabolic and structural genes by remodeling H3K27ac depositions in related enhancer regions. Integration of the ERRγ ChIP-seq data with published hiPSC-CM datasets demonstrated that approximately 50% of super-enhancer regions overlapped ERRγ peaks, suggesting ERR is involved in the maintenance and function of cardiac super-enhancer regions. Corresponding motif analyses and intersection analyses with published GATA4 ChIP-seq datasets suggested that ERRγ cooperates with GATA4 on many structural, but not metabolic targets. Specifically, binding sites for both factors are often co-localized around cardiac contractile, ion channel, and Ca
2+
handling genes, including
TNNI3, MYBPC3, MYH7, TTN, KCNQ1, and RYR2
. In contrast, ERRγ sites on target genes involved in mitochondrial maturation such as
ATP5B, COX4I1, CPT1B
, and
FABP3
lacked GATA4 sites. Experiments using luciferase reporters demonstrated functional cooperation between ERRγ and GATA4 that depends on the known ERR coactivator, peroxisome proliferator-activated receptor gamma co-activator 1alpha (PGC-1α). We conclude that the ERR cooperates with GATA4 to drive cardiac structural gene transcription during cardiac differentiation, whereas ERR regulates genes involved in energy metabolism independent of GATA4. Activation of ERR signaling could prove to be an effective tool to drive hiPSC-CM maturation.
Abstract 223: Yap and Its Homolog Wwtr1 Are Regulators of Myofibroblast Activation Following Ischemic Injury
Authors: Flinn Michael A , Alencar Victor , Patterson Michaela , Omeara Caitlin
Published: October 2020

Introduction:
Following ischemic injury in adult mammals, cardiac fibroblasts differentiate into myofibroblasts and promote secretion of matrix fibers. Myofibroblast activation is critical for initial scar formation and preventing heart rupture, however, extended activity can lead to heart failure progression. Thus, there is a need to identify the mechanisms mediating persistent activation of myofibroblasts to prevent excessive fibrosis and adverse cardiac remodeling. Here we demonstrate that Hippo-Yap pathway offers a target for modulating myofibroblast activation and thus the fibrotic response.

Methods and Results:
We tested the hypothesis that Yap and its homolog Wwtr1 (known as ‘Taz’) are regulators of myofibroblast activation following ischemic injury. We implemented a Cre-lox system whereby Yap alone or both Yap and Wwrt1 were depleted using an inducible Cre expressed under a myofibroblast specific promoter (
Postn
MCM
). Following permeant ligation of the left anterior descending artery in adult mice, we found that myofibroblast depletion of Yap alone resulted in a significant reduction in left ventricular dilation 28 days post injury (dpi) and decreased proliferation of scar associated cells. Strikingly, myofibroblast specific depletion of Yap and one copy of Wwrt1 resulted in further attenuation of left ventricular dilation as well as improved fractional shortening and ejection fraction at 28 and 60 dpi. Histological assessment revealed that depletion of both Yap and Wwrt1 resulted in greater than 50% reduction in scar size (by midline) at 60 dpi. RNAseq of whole hearts collected at 4 dpi suggested that Hippo-Yap pathway expression specifically in myofibroblasts facilitates immune cell recruitment in the heart. Collectively These data illustrate a role for Hippo-Yap signaling mediating myofibroblast activity and immune cell coordination following injury and therefore cardiac fibrosis and remodeling.

Conclusions:
Our data demonstrate that endogenous Yap and Wwrt1 deletion in myofibroblasts suppresses the fibrotic response, mediates inflammation, and improves cardiac function after ischemic injury. These results therefore offer a regulatory pathway that can be targeted therapeutically to prevent progressive heart failure.
Abstract 224: A Deep Learning Approach to Left-Ventricular Chamber Quantification for Fully Automated Three Dimensional Strain Analysis in Cardiotoxicity
Authors: Baldwin Bryant M , Joseph Shane , Zhong Xiaodong , Kakish Ranya , Revere Cherie , Poosarla Teja , Malozzi Christopher M , McQuiston Samuel , Cohen Michael V , Kar Julia
Published: October 2020

This study investigated MRI and semantic segmentation-based deep-learning (SSDL) automation for left-ventricular chamber quantifications (LVCQ) and low longitudinal strain (LLS) determination, thus eliminating user-bias by providing an automated tool to detect cardiotoxicity (CT) in breast cancer patients treated with antineoplastic agents. Displacement Encoding with Stimulated Echoes-based (DENSE) myocardial images from 26 patients were analyzed with the tool’s Convolution Neural Network with underlying Resnet-50 architecture. Quantifications based on the SSDL tool’s output were for LV end-diastolic diameter (LVEDD), ejection fraction (LVEF), and mass (LVM) (see figure for phase sequence). LLS was analyzed with Radial Point Interpolation Method (RPIM) with DENSE phase-based displacements. LVCQs were validated by comparison to measurements obtained with an existing semi-automated vendor tool (VT) and strains by 2 independent users employing Bland-Altman analysis (BAA) and interclass correlation coefficients estimated with Cronbach’s Alpha (C-Alpha) index. F1 score for classification accuracy was 0.92. LVCQs determined by SSDL and VT were 4.6 ± 0.5 vs 4.6 ± 0.7 cm (C-Alpha = 0.93 and BAA = 0.5 ± 0.5 cm) for LVEDD, 58 ± 5 vs 58 ± 6 % (0.90, 1 ± 5%) for LVEF, 119 ± 17 vs 121 ± 14 g (0.93, 5 ± 8 g) for LV mass, while LLS was 14 ± 4 vs 14 ± 3 % (0.86, 0.2 ± 6%). Hence, equivalent LV dimensions, mass and strains measured by VT and DENSE imaging validate our unique automated analytic tool. Longitudinal strains in patients can then be analyzed without user bias to detect abnormalities for the indication of cardiotoxicity and the need for therapeutic intervention even if LVEF is not affected.
Abstract 225: Coordinated Transcriptome and Cell State Dynamics of Non-myocytes in Heart Regeneration
Authors: Ma Hong , Liu Ziqing , Yang Yuchen , Feng Dong , Dong Yanhan , Rivis Andrea A , Garbutt Tiffany A , Hu Zhiyuan , Wang Li , Cooper Cynthia D , Li Yun , Joshua Welch , Qian Li , Liu Jiandong
Published: October 2020

Cardiac regeneration occurs primarily through proliferation of existing cardiomyocytes, yet the regenerative response also involves complex interactions between distinct cardiac cell types including not only cardiomyocytes, but also non-cardiomyocytes (nonCMs). However, the subpopulations, distinguishing molecular features, cellular functions, and intercellular interactions of nonCMs in heart regeneration remain largely unexplored. Using the LIGER algorithm, we assembled an atlas of cell states from 61,977 individual nonCM scRNA-seq profiles isolated at multiple time points during heart regeneration in both wildtype and mutant fish. This analysis revealed extensive nonCM cell diversity, including multiple macrophage, fibroblast and endothelial subpopulations with unique spatiotemporal distributions and cooperative interactions during the process of cardiac regeneration. Genetic and pharmacological perturbation of macrophage functional dynamics compromised interactions among nonCM subpopulations, reduced cardiomyocyte proliferation, and caused defective cardiac regeneration. Furthermore, we developed a computational algorithm called Topologizer to map the topological relationships and dynamics of nonCMs during heart regeneration. We uncovered dynamic transitions between macrophage functional states and identified factors involved in mRNA processing and transcriptional regulation associated with the transition. Together, our single-cell transcriptomic analysis of nonCMs during cardiac regeneration provides a blueprint for interrogating the molecular and cellular basis of cardiac regeneration.
Abstract 227: Lack of Beta-carotene 9’,10’ Diooxygenase (Bco2) Impacts Cardiac Postnatal Growth and Energy Metabolism in Female Mice
Authors: Holloway Chelsee , Kim Youn-kyung , Zhong Guo , Isoherranen Nina , Quadro Loredana
Published: October 2020

As the postnatal heart grows it also undergoes critical metabolic adaptations to use mainly lipids rather than carbohydrates as energy-providing substrates. Abnormal growth patterns and lack of metabolic flexibility are pathological responses of the heart that lead to cardiovascular diseases (CVD). Carotenoids, such as β-carotene - the most abundant dietary precursor of vitamin A - have been linked to the prevention of CVD. In the heart, β-carotene can be metabolized to retinoids and other apo-carotenoids by the action of β-carotene 9’,10’-dioxygenase (BCO2). We found that

Bco2
-/-

female mice have significantly smaller hearts than wild-type (WT), pre- and post-puberty. Echocardiography revealed a significant increase in aortic blood flow velocity with no change in overall cardiac output. This analysis also suggests that the mutant has a smaller left ventricle with thinner walls. In addition, expression of cardiac foetal genes
(BNP, α-Skeletal actinin
and
β-MyHc
), typically upregulated during embryogenesis or under pathological conditions in the adult, were significantly increased in the mutant.

Bco2
-/-

females showed impaired exercise capacity, displaying a ~40% reduction in running distance compared to WT. These data indicate functional cardiac defects in the mutant under stress conditions.

Bco2
-/-

females also displayed impaired cardiac metabolic flexibility. We found that

Bco2
-/-

mice have reduced
Pdk4
expression and thus enhanced PDH activity. Cardiac expression of
Glut1
and
Mct1,
key glucose and lactate transporters
,
were significantly higher in

Bco2
-/-

females, while triglyceride levels were lower in the heart and serum. These findings suggest preferential use of glucose as a cardiac energy source. Interestingly, the mutants had reduced cardiac retinoic acid levels (by LC-MS analysis) compared to WT. Retinoic acid upregulates
Pdk4
, thus we postulated that retinoic acid deficiency contributes to loss of cardiac metabolic flexibility in

Bco2
-/-

females. Ongoing studies aim to confirm this hypothesis and determine if altered retinoic acid homeostasis is also linked to impaired postnatal heart growth in mutant mice. These findings give new insights into a potential role of the β-carotene metabolic pathway in regulating adult heart function.
Abstract 228: Inflammasome-Primed Neutrophils Return to the Bone Marrow to Stimulate Leukocytosis Following Myocardial Infarction
Authors: Nagareddy Prabhakara R , Sreejit Gopalkrishna , Lee Man K , Athmanathan Baskaran , Quaife-Ryan Greg A , Abdel-Latif Ahmed , Porrello Enzo R , Murphy Andrew
Published: October 2020

Myocardial infarction (MI) triggers myelopoiesis resulting in heightened number of neutrophils in the circulation. However, the mechanism that sustain their number and recruitment to the infarcted heart are unclear. Here, we show that in a mouse model of MI (permanent ligation of LAD), neutrophils are rapidly recruited to the infarct, where they release specific alarmins, S100A8 and S100A9. These alarmins acting either in an autocrine or paracrine manner, primed the Nod Like Receptor (NLR) family Pyrin Domain Containing 3 (Nlrp3) inflammasome in naïve neutrophils via their interaction with the Toll Like Receptor (TLR) 4. The interaction did not result in the release of IL1β systemically. However, the primed neutrophils, loaded with pro-interleukin-1β (IL-1β) returned to the bone marrow (BM) in a CXCR4 (C-X-C-motif chemokine receptor 4)- dependent manner. While at the BM, the primed-neutrophils released IL-1β through Gasdermin D pores and, stimulated granulopoiesis in a cell-autonomous manner. Strategies aimed at preventing the Nlrp3 inflammasome-priming or re-entry of the primed neutrophils to the BM dampened MI-induced granulopoiesis and markedly improved cardiac function. In subjects with acute ST-elevation myocardial infarction (STEMI), the number of neutrophils in the circulation increased both at the time of admission and following revascularization. Most importantly, patients with higher peak neutrophil counts demonstrated significantly higher incidence of major adverse cardiovascular events (MACE) during the one year follow up period. Similar to mouse data, the plasma levels of IL-1β did not change in STEMI patients at any time. However, the circulating neutrophils carried greater amounts of pro-IL-1β confirming our mouse data that granulopoiesis is likely not induced by systemic but locally delivered IL-1β by reverse migrating neutrophils. These data reveal a new paradigm of how circulatory cells establish direct communication between organs by delivering signaling molecules directly at the sites of action rather through systemically.
Abstract 237: Noncanonical WNT Activation in Human Right Ventricular Heart Failure
Authors: Edwards Jonathan , Brandimarto Jeffrey A , Hu Dong-Qing , Yucel Nora , Li Li , Jeong Sunhye , Bedi Kenneth C , Hwang Hyun Tae , Zhao Mingming , Margulies Kenneth B , Bernstein Daniel , Reddy Sushma , Arany Zoltan
Published: October 2020

Background:
Drivers of adaptive and pathologic human right ventricular (RV) remodeling are poorly defined, but there is some evidence that WNT signaling contributes to these processes.

Methods:
Using a candidate gene approach, we sought to identify genes specifically expressed in RV failure by assessing the expression of 28 WNT-related genes in the RVs of three groups: human explanted nonfailing donor hearts (NF, n = 29), pre-transplant cardiomyopathy hearts with preserved RV function (pRV, n = 78), and pre-transplant cardiomyopathy hearts with RV failure (RVF, n = 35).

Results:
We identified the noncanonical WNT receptor ROR2 as transcriptionally strongly upregulated in RVF compared to pRV and NF (p < 0.05, adjusted for multiple hypothesis testing using Benjamin-Hochberg). By western blot, ROR2 protein expression correlated to mRNA expression (R
2
= 0.41, p = 8.1 x 10-18) and increased linearly with higher right atrial to pulmonary capillary wedge ratio in RVF (R
2
=0.40, p = 3.0 x 10-5). Using trichrome, immunohistochemistry, and RNAscope, we identified preferential ROR2 mRNA and protein expression in fibrotic regions by both cardiomyocytes and noncardiomyocytes. High ROR2 expression correlated with increased expression of a known ROR2 target, the WNT/Ca2+ responsive protease calpain and its target FLNA (R
2
0.25 and 0.67, respectively, p < 0.05), as well as with increased relative phosphorylation of FLNA (R
2
0.62, p < 0.05), a marker of its activation by ROR2. Finally, mice with a range of RVF severity, generated by pulmonary artery constriction, demonstrated a robust increase in ROR2 and FLNA expression compared to sham-treated animals, and ROR2 and FLNA expression correlated strongly with each other (R
2
0.89, p = 6.9 x 10-5) and with increasing RV weight normalized to body weight (R
2
~0.8, p < 0.001, both).

Conclusion:
ROR2 expression in the RV is dramatically increased in humans and mice with severe RVF, and ROR2 expression correlates with worsening RV hemodynamics, calpain expression, FLNA cleavage, and FLNA phosphorylation. Taken together, the data reveal the robust activation of noncanonical WNT signaling in human RVF, and identify ROR2-mediated cytoskeleton protein cleavage as a potential novel target of human pathologic RV remodeling.
Abstract 241: Retinoic Acid Promotes Metabolic Maturation of Human Embryonic Stem Cell-derived Cardiomyocytes
Authors: Miao Shumei , Fang Xing , Wang Xiaoxiao , Ye Lingqun , Yang Jingsi , Ni Xuan , Zhao Dandan , Wu Hongchun , Han Xinglong , Yu Miao , Li Lei , Shen Zhenya , Lei Wei , Zhao Zhen-Ao , Hu Shijun
Published: October 2020

Cardiomyocytes differentiated from human embryonic stem cells (hESCs) represent a promising cell source for heart repair, disease modeling and drug testing. However, improving the differentiation efficiency and maturation of hESC-derived cardiomyocytes (hESC-CMs) is still a major concern. Retinoic acid (RA) signaling plays multiple roles in heart development, and studies on RA can provide clues for understanding cardiomyocyte differentiation and maturation. In this study, we studied the roles of RA during cardiomyocyte differentiation and maturation, systematically. After adding RA at different stages of cardiomyocyte differentiation, we compared the efficiency of differentiation by quantitative real-time PCR and flow cytometry. We found that RA treatment at the lateral mesoderm stage (days 2-4) significantly improved cardiomyocyte differentiation, as evidenced by the upregulation of
TNNT2, NKX2.5
and
MYH6
on day 10 of differentiation. In addition, flow cytometry showed that the proportion of differentiated cardiomyocytes in the RA-treated group was significantly higher than that in control group. Furthermore, RA was added at different time intervals after purification to induce cardiomyocyte maturation. Our results demonstrated that RA treatment on days 15-20 increased cardiomyocyte area, sarcomere length, multinucleation and mitochondrial copy number, and promoted RNA splicing switch. Importantly, RA-treated cardiomyocytes showed decreased glycolysis and enhanced mitochondrial oxidative phosphorylation, with the increased utilization of fatty acid and exogenous pyruvate but not glutamine. In conclusion, our data indicated that RA treatment at an early time window (days 2-4) promotes the efficiency of cardiomyocyte differentiation and that RA treatment post beating (days 15-20) promotes cardiomyocyte metabolic maturation. The biphasic effects of RA provide new insights for improving cardiomyocyte differentiation and quality.