bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of Nutritional Composition in 23 Kinds of Edible Fungi
Authors: Yu Qiannan , Guo Meijuan , Zhang Bin , Wu Hao , Zhang Yan , Zhang Lantian
Published: October 2020

To explore the differences in the main nutritional composition of 23 kinds of common edible fungi in the market, the moisture, ash, protein, fat, dietary fiber, carbohydrates, polysaccharides, and energy were analyzed using national standard methods. The results showed that the 23 kinds of edible fungi varied greatly in nutritional composition. Based on dry weight, the moisture content was 6.9∼15.5 g/100 g, the ash content ranged from 1.3 to 10.1 g/100 g, the protein content ranged from 8.5 to 36.9 g/100 g, the fat content was 0.5∼3.9 g/100 g, the dietary fiber content was between 14.4∼70.2 g/100 g, the carbohydrate content is 0.5∼37.3 g/100 g, the polysaccharide content was 2.1∼8.3 g/100 g, and the energy is about 751∼1322 100 g/kJ. All the 23 kinds of edible fungi can be regarded as high-protein low-fat foods, which have their own advantages in terms of nutritional value. This study provides reference for people to use edible fungi in a more scientific and reasonable way.
Optimization and Simulation of Controller Area Network Communication Model Based on Industrial Internet of Things Platform
Authors: Lin Haifeng , Du Lin
Published: October 2020

The Internet of Things (IoT) is an information carrier based on network communication, and it can make all general physical objects which can be independently addressed from an interconnected network. Controller Area Network (CAN) is a kind of serial communication network which can effectively support distributed control or real-time control. This paper proposes the design of an IoT-oriented universal CAN bus. In order to build a complete CAN communication simulation model, message sending and message receiving models are constructed, respectively. The simulation experiment proves that the IoT-oriented network communication model designed in this paper not only has low power consumption, but it can also solve the front-end compatibility problems caused by different communication protocols.
Adipose Tissue-Derived Stromal Cells Induce a Highly Trophic Environment While Reducing Maturation of Monocyte-Derived Dendritic Cells
Authors: Juhl Morten , Follin Bjarke , Gad Monika , Larsen Jesper , Kastrup Jens , Ekblond Annette
Published: October 2020

Allogeneic cell-based therapies using adipose tissue-derived stromal cells (ASCs) offer an off-the-shelf alternative to autologous therapy. An underlying assumption is that ASC can modulate the immune response of the recipient. However, in vitro models are required to explore and identify cell interactions and mechanisms of action, to ensure sufficient and sustained effects, and to document these. In this study, we shed light on the effect of ASC manufactured for clinical use on monocyte-derived dendritic cells and an inflammatory microenvironment. ASCs were isolated from healthy voluntary donors, expanded using a human platelet lysate in bioreactors, and cryopreserved as per clinical use. Monocyte-derived dendritic cells were generated by isolation of monocytes and differentiation with GM-CSF and IL-4. Dendritic cells were cocultured with different ratios of ASC and matured with LPS and IFN-γ. Dexamethasone was included as an immunosuppressive control. Dendritic cells were analyzed by flow cytometry for CD11c, CD40, CD80, CD83, CD86, PD-L1, and HLA-DR, and supernatants were analyzed for FGF2, HGF, IL-10, IL-12p70, LIF, MIF, PDGF, PlGF, and IDO. Reduced expression of maturation markers was observed on ASC-treated dendritic cells, while high levels of PD-L1 were maintained. Interestingly, the expression of CD83 was elevated. Escalating ratios of ASC did not affect the concentration of IL-10 considerably, whereas the presence of IL-12 was reduced in a dose-dependent manner. Besides offsetting the IL-12/IL-10 balance, the concentrations of IDO and MIF were elevated in cocultures. Concentrations of FGF2, HGF, LIF, and PIGF were high in ASC cocultures, whereas PDGF was depleted. In a robust coculture model, the addition of ASC to dendritic cells inhibited the dendritic maturation substantially, while inducing a less inflammatory and more tolerogenic milieu. Despite the exposure to dendritic cells and inflammatory stimuli, ASC resulted in supernatants with trophic factors relevant for regeneration. Thus, ASC can perform immunomodulation while providing a regenerative environment.
Ovarian Cancer Metastasis to the Breast: A Case Report and Review of the Literature
Authors: Caruso Giuseppe , Musacchio Lucia , Santangelo Giusi , Palaia Innocenza , Tomao Federica , Di Donato Violante , Perniola Giorgia , Salutari Vanda , Benedetti Panici Pierluigi
Published: October 2020

Although ovarian cancer often presents as a widespread disease, metastases to the breast and/or axillary lymph nodes are a very rare event, accounting for only 0.03–0.6% of all breast cancers. Its early recognition and accurate distinction from primary breast cancer are of crucial importance to choose an adequate systemic therapy over unnecessary surgeries. We presented the case of a 53-year-old woman who was diagnosed with breast metastases 2 years after the diagnosis of advanced primary serous ovarian cancer. The patient underwent primary cytoreductive surgery and platinum-based chemotherapy in combination with bevacizumab, followed by bevacizumab maintenance for 18 months. After 2 years of negative follow-ups, the disease unexpectedly spread to the left breast and axillary lymph nodes. No axillary lymph node dissection or breast surgery was performed. The patient received axillary radiotherapy and multiple chemotherapy lines: gemcitabine/cisplatin, liposomal doxorubicin, topotecan, olaparib/cediranib, paclitaxel, and cisplatin. Unfortunately, none of these treatments improved her prognosis and she died 3 years after the disease recurrence. Ovarian cancer metastasis to the breast reveals a disseminated disease with a poor prognosis. Currently, no valid treatment options are available as the disease shows multidrug chemoresistance. In the era of precision medicine, the characterization of genetic and molecular markers may play a role in offering new promising targeted therapies.
Blind Pouch Syndrome-Associated Enterolithiasis Successfully Treated with Colonoscopy
Authors: Kubo Kimitoshi , Suzuoki Masato , Kimura Noriko , Maiya Norishige , Matsuda Soichiro , Tsuda Momoko , Ohara Masanori , Kato Mototsugu
Published: October 2020

Enterolithiasis associated with blind pouch syndrome secondary to functional end-to-end anastomosis is rare, and its endoscopic and radiological features remain poorly described. A 72-year-old woman was admitted to our hospital for abdominal pain and difficulty defecating. Colonoscopy (CS) with Gastrografin revealed a 10 × 8 cm calculus, an anastomotic ulcer, a blind pouch, and an end-to-end anastomosis in the transverse colon. The calculus was successfully crushed and removed with snares and alligator forceps through CS during the ensuing 4-day period. To our knowledge, this is the first report describing the endoscopic and radiological features of blind pouch syndrome-associated enterolithiasis successfully treated with CS.
Misinterpretation of Hereditary Breast Cancer Risk and Its Association with Information Sharing Motives among Women at Low Likelihood of Carrying a BRCA1/2 Mutation
Authors: Zhao Jingsong , McBride Colleen M. , Guan Yue
Published: October 2020

Purpose: In this brief report, we ask whether women’s interpretation of breast cancer risk based on their low likelihood of carrying a BRCA1/2 mutation is associated with their information-sharing behavior, and whether misinterpretation is associated with motives for sharing the result. Methods: Women in mammography clinics who completed a brief family history assessment and deemed to be at low likelihood of carrying a BRCA1/2 mutation were asked to complete a 1-time online survey between June 2016 and January 2017. Results: One-third (44/148) of women shared their family history screen result with someone in their social network. Result information was shared largely with a first-degree female relative to express feelings of relief (77%, 33/43). There were no differences in likelihood of sharing based on breast cancer risk interpretation. However, women who misinterpreted the implications of the result for general breast cancer risk reported more motives to share the result with their social network than those who accurately interpreted their breast cancer risk. Conclusions: As family history-based screening for hereditary breast cancer is broadly implemented, the communication needs of the majority of women who will be unlikely of carrying a BRCA1/2 mutation must be considered. The motives of women who misinterpreted the implications of this result for breast cancer risk suggest the possibility that miscommunication could be spread to the broader family network.
Fetal Surgery in the Primate 4.0: A New Technique 30 Years Later
Authors: Perrone Erin E. , Galganski Laura A. , Tarantal Alice F. , Olstad Katie J. , Treadwell Marjorie C. , Berman Deborah R. , Jarboe Marcus D. , Mychaliska George B. , Farmer Diana L.
Published: October 2020

Introduction: Open fetal surgery requires a hemostatic hysterotomy that minimizes membrane separation. For over 30 years, the standard of care for hysterotomy in the gravid uterus has been the AutoSuture Premium Poly CS*-57 stapler. Objective: In this study, we sought to test the feasibility of hysterotomy in a rhesus monkey model with the Harmonic ACE®+7 Shears. Methods: A gravid rhesus monkey underwent midgestation hysterotomy at approximately 90 days of gestation (2nd trimester; term = 165 ± 10 days) using the Harmonic ACE®+7 Shears. A two-layer uterine closure was completed and the dam was monitored by ultrasound intermittently throughout the pregnancy. At 58 days after hysterotomy (near term), a final surgery was performed to evaluate the uterus and hysterotomy site. Results: A 3.5-cm hysterotomy was completed in 2 min 7 s. The opening was hemostatic and the membranes were sealed. Immediately after closure and throughout the pregnancy, ultrasound revealed intact membranes without separation and normal amniotic fluid levels. At term, the scar was well healed without signs of thinning or dehiscence. Conclusions: The Harmonic ACE®+7 Shears produced a hemostatic midgestation hysterotomy with membrane sealing in the rhesus monkey model. Importantly, healing was acceptable.
cinema-caracol de Luis Tróchez Tunubalá
Authors: Felipe Marcos Aurélio
Published: October 2020

Este estudo analisa o documentário Na Misak (2018), do diretor indígena Luis Tróchez Tunubalá, do Cauca colombiano. Situando-o como filme-prismático, no marco do cinema indígena e do colonialismo reiterativo em Abya Yala, seu objetivo é investigar a tessitura fílmica a partir da perspectiva indígena sobre a construção da indianidade no contexto urbano contemporâneo. Utilizou-se o método comparado a partir de cotejos baseados na série histórica, ponto-contraponto, constelações e abordagem prismática. Entre os estudos antropológicos, históricos e de cinema, observou-se o cruzamento da escritura cinematográfica contracolonial, ensaística e autorreflexiva com dimensões da cosmologia Misak, a partir do qual o pensamento crítico se condensa no sistema fílmico, na experiência histórica e na performance corporal, artefatos que confrontam o(s) colonialismo(s) com outra história do contato.
Subjectivité de la représentation du genre littéraire chez Jouri Tynianov et Mikhaïl Bakhtine: essai de définition et étude de textes
Authors: Garziano Svetlana
Published: October 2020

Afin d’approfondir l’examen de la nature du genre littéraire et comme le présent volume porte en partie sur les questions du formalisme russe, cet article s’intéressera à l’expression de la subjectivité appliquée aux genres littéraires telle qu’elle est exposée dans quelques textes fondamentaux écrits par deux grands théoriciens du verbe russe : d’une part, il s’agit de Jouri Tynianov, l’un des principaux acteurs de l’OPOÏAZ, le fondateur du formalisme russe avec Victor Chklovski, Roman Jacobson et Boris Eïkhenbaum, et, d’autre part, c’est Mikhaïl Bakhtine qui a entamé son cheminement intellectuel au moment où « le formalisme était au faîte de sa gloire ».
Risk Factors and Timing of Acute Myocardial Infarction Associated With Pregnancy: Insights From the National Inpatient Sample
Authors: Balgobin Courtney A. , Zhang Xiaoyue , Lima Fabio V. , Avila Cecilia , Parikh Puja B. , Yang Jie , Stergiopoulos Kathleen
Published: October 2020

Background

Pregnancy increases the risk of acute myocardial infarction (AMI). The purpose of this study was to examine timing and risk factors for AMI in pregnancy and poor outcome.

Methods and Results

National Inpatient Sample (2003–2015) was screened in pregnancy, labor and delivery, and postpartum. There were 11 297 849 records extracted with 913 instances of AMI (0.008%). One hundred eleven (12.2%) women experienced AMI during labor and delivery, 338 (37.0%) during pregnancy and most during the postpartum period (464; 50.8%). The prevalence of AMI in pregnancy has increased (
P
=0.0005). Most major adverse cardiovascular and cerebrovascular events occurred in the postpartum period (63.5%). Inpatient mortality was 4.5%. Predictors of AMI include known coronary artery disease (odds ratio [OR], 517.4; 95% CI, 420.8–636.2), heart failure (OR, 8.2; 95% CI, 1.9–35.2), prior valve replacement (OR, 6.4; 95% CI, 2.4–17.1), and atrial fibrillation (OR, 2.7; CI, 1.5–4.7;
P
<0.001). Risk factors of traditional atherosclerosis including hyperlipidemia, obesity, tobacco history, substance abuse, and thrombophilia were identified (
P
<0.001). Gestational hypertensive disorders (eclampsia OR, 6.0; 95% CI, 3.3–10.8; preeclampsia OR, 3.2; 95% CI, 2.5–4.2) were significant risk factors in predicting AMI. Risk factors associated with major adverse cardiovascular and cerebrovascular events included prior percutaneous coronary intervention (OR, 6.6; 95% CI, 1.4–31.2) and pre‐eclampsia (OR, 2.3; 95% CI, 1.3–3.9).

Conclusions

AMI is associated with modifiable, nonmodifiable, and obstetric risk factors. These risk factors can lead to devastating adverse outcomes and highlight the need for risk factor modification and public health resource initiatives toward the goal of decreasing AMI in the pregnant population.
Regional Right Ventricular Abnormalities Implicate Distinct Pathophysiological Conditions in Patients With Chronic Thromboembolic Pulmonary Hypertension
Authors: Moriyama Hidenori , Kawakami Takashi , Kataoka Masaharu , Hiraide Takahiro , Kimura Mai , Endo Jin , Kohno Takashi , Itabashi Yuji , Seo Yoshihiro , Fukuda Keiichi , Murata Mitsushige
Published: October 2020

Background

Right ventricular (RV) dysfunction is a prognostic factor for cardiovascular disease. However, its mechanism and pathophysiology remain unknown. We investigated RV function using RV‐specific 3‐dimensional (3D)‐speckle‐tracking echocardiography (STE) in patients with chronic thromboembolic pulmonary hypertension. We also assessed regional wall motion abnormalities in the RV and chronological changes during balloon pulmonary angioplasty (BPA).

Methods and Results

Twenty‐nine patients with chronic thromboembolic pulmonary hypertension who underwent BPA were enrolled and underwent right heart catheterization and echocardiography before, immediately after, and 6 months after BPA. Echocardiographic assessment of RV function included both 2‐dimensional‐STE and RV‐specific 3D‐STE. Before BPA, global area change ratio measured by 3D‐STE was significantly associated with invasively measured mean pulmonary artery pressure and pulmonary vascular resistance (
r
=0.671 and
r
=0.700, respectively). Dividing the RV into the inlet, apex, and outlet, inlet area change ratio showed strong correlation with mean pulmonary artery pressure and pulmonary vascular resistance before BPA (
r
=0.573 and
r
=0.666, respectively). Only outlet area change ratio was significantly correlated with troponin T values at 6 months after BPA (
r
=0.470), and its improvement after BPA was delayed compared with the inlet and apex regions. Patients with poor outlet area change ratio were associated with a delay in RV reverse remodeling after treatment.

Conclusions

RV‐specific 3D‐STE analysis revealed that 3D RV parameters were novel useful indicators for assessing RV function and hemodynamics in pulmonary hypertension and that each regional RV portion presents a unique response to hemodynamic changes during treatment, implicating that evaluation of RV regional functions might lead to a new guide for treatment strategies.
Abstract 202: In Vitro Effect of Trisulfate Disaccharide on ICa (L-type), INa, IK, and the Na/Ca Exchange Current Recorded From Rat Ventricular Myocytes by Patch Clamp
Authors: Vasques Enio R , Nader Helena , Tersariol Ivarne , Carlos Godoy
Published: October 2020

Background:
Ion channels are pharmacological targets for antiarrhythmic action, and drugs currently used for this purpose are generally not specific to a site of action and may act on several channels and even trigger proarrhythmic phenomena. Trisulfate disaccharide (TD) is an heparin fragment known to act on the sodium calcium exchanger (NCX), reducing intracellular calcium in overload situations and reversing arrhytmias, but its action on other ionic currents is unknown.

Objective:
To evaluate by patch clamp the action of TD at different concentrations in NCX and ionic currents in situations of intracellular calcium overload.

Materials and Methods:
Adult rat myocytes were obtained from a sample from ventricles. Currents were measured using the whole-cell variant of the patch clamp method. Creation of voltage clamp pulses and data acquisition was controlled by a computer with pClamp software. Peak inward current amplitude was measured for ion currents. For Na/Ca exchange current a ramp voltage protocol was employed. Three different concentrations of Cai (300nM, 400nM and 600nM) were used in separate experiments. One drug concentration was applied per cell (10, 30 and 100 micromolar each). The current sensitive to 5mM nickel was taken as the Na/Ca exchange current. The effects of TD on the INa, L-type Ca, and the potassium currents, transiente outward current (Ito), inwardly rectifying potassium current (IK1), and sustained current (Isus) recorded from adult rat ventricular myocytes were also examined in the same conditions.

Results:
TD concentration-dependently increased the inward Na/Ca exchange current in all intracellular calcium concentration. The effects of TD on the INa, L-type Ca, and the potassium currents, Ito, IK1 and Isus was associated with less than 30% mean reduction on any current at the highest concentration of TD tested (100 micromolar) and still below the positive block controls for different channels that is above 40% block.

Conclusion:
TD acts on NCX under different concentrations used, without affecting other ionic currents, suggesting specificity in the mechanism of action and possibly not exerting a pro-arrhythmic activity, this effect being desirable for its possible use in reversal of cardiac arrhythmias.
Abstract 207: A Catalogue of the Pathogenic Mutations of LDL Receptor Gene in Japanese Familial Hypercholesterolemia
Authors: Tada Hayato , Usui Soichiro , Kawashiri Masa-aki , Takamura Masayuki , Harada-shiba Mariko
Published: October 2020

Background:
Familial hypercholesterolemia (FH), mainly caused by LDL receptor gene mutation is attracting a lot of attention associated with the development of novel therapies. However, rare mutations tend to be ethnicity-specific, hence, it is always difficult to determine pathogenicity of LDL receptor gene mutation in a single individual in a certain ethnicity. Currently, little data exist on the pathogenic mutations of LDL receptor in Japanese familial hypercholesterolemia (FH).

Objective:
We aimed to catalogue the pathogenic mutations of LDL receptor gene in the 2 major Japanese FH-care centers (Kanazawa University and National Cerebral and Cardiovascular Center Research Institute), where genetic testing of FH has been performed centrally upon requests from institutes all over Japan during more than past two decades.

Methods:
796 FH subjects from 472 families who had nonsynonymous mutations in LDL receptor gene were included in this study. Genetic mutations were analyzed for mutations by Sanger sequencing as well as by multiplex ligation probe dependent amplification (MLPA) technique for large rearrangements. Pathogenic mutations were defined either as 1) protein truncated variants, 2) registered as pathogenic in Clinvar, or Human Gene Mutation Database (HGMD), or meet the criteria of American College of Medical Genetics and Genomics (ACMG) guideline, or 3) CADD-score > 10.

Results:
We found 138 different mutations. Among them, 132 mutations were considered as pathogenic, including 19 large rearrangement mutations. Whereas, 6 missense mutations were classified as variants of unknown significance (VUS). A single mutation accounted for as much as 41 % of the FH subjects recruited from Kanazawa University mainly due to founder gene effect, whereas, many singleton mutations were found from National Cerebral and Cardiovascular Center Research Institute located in Osaka.

Conclusions:
We provided the largest catalogue of pathogenic mutations of LDL receptor gene in Japanese FH. This could aid to determine the pathogenicity of the LDL receptor genetic mutations not only in Japanese but also in other ethnicities.
Abstract 210: Reprogramming Mouse Fibroblasts Into Cardiac Progenitor Cells Using Crispr Technique Targeting Endogenic Genes
Authors: Wang Yi-gang , Jiang Lin , Liang Jialiang , Huang Wei , Paul Christian
Published: October 2020

Background and Objective:
CRISPR tools that allow for precise manipulation of individual loci have not been used in generation of
i
nduced
c
ardiac
p
rogenitor
c
ells (
iCPC
s). This study was designed to determine the feasibility and effectiveness of reprogramming fibroblasts into iCPC using CRISPR activation (CRISPRa) system.

Methods:
Tail-tip fibroblasts (TTFs) were isolated from Nkx2-5 cardiac enhancer GFP reporter mice. A gRNA pool targeting 17 progenitor genes was synthesized and transduced with dCas9-VP64 lentivirus into TTFs (
Fig.1A
). The phenotype of iCPCs was then characterized by immunostaining and FACS of progenitor markers. Finally, the cardiac-lineage differentiation potential of iCPCs was determined by immunostaining and electrophysiological assay under defined induction mediums.

Results:
iCPCs with GFP expression were formed in TTFs after transduction of CRISPRa targeting
Isl1, Gata4, Baf60c, Tbx5
and
Nkx2-5
(Fig.1B), while GFP was not activated by control virus. Cardiac progenitor markers were activated in iCPCs as shown by immunostaining (Fig.1C). The generation efficiency of Flk1-postive iCPCs induced by CRISPRa was ~60% as showed by FACS. iCPCs can be differentiated into cardiomyocytes as identified by immunostaining of cardiac-specific markers (Fig.1D). The iCPC-derived cardiomyocytes displayed spontaneous beating and showed cardiac action potentials (Fig.1E).

Conclusion:
The CRISPRa system is an efficient and specific way to generate iCPCs, which could provide a novel source of cells for cardiac regenerative medicine. Conversion of fibroblasts into cardiac progenitors poses a novel therapeutic option for repair of myocardial infarction.
Abstract 211: Long Noncoding RNA Myocardial Infarction Associated Transcript, MIAT Exacerbates Maladaptive Cardiac Remodeling by Functioning as a Competing Endogenous RNA of MicroRNA-150 During Myocardial Infarction
Authors: Aonuma Tatsuya , Moukette Bruno , Kim Il-man
Published: October 2020

Cardiac injury is accompanied by dynamic changes in the expression of noncoding RNAs such as microRNAs (miRs) and long noncoding RNAs (lncRNAs) that regulate target genes. We previously reported that β
1
-adrenergic receptor/β-arrestin1-responsive miR-150 plays a vital cardioprotective role in myocardial infarction (MI) via decreasing cardiac cell apoptosis (Fig. A). MiR-150 is regulated by MI-Associated Transcript (MIAT), a lncRNA that directly interacts with miR-150 and functions as its competing endogenous RNA
in vitro
. Gain-of-function single nucleotide polymorphisms in MIAT are associated with increased risk of MI in humans, and MIAT is upregulated in post-MI hearts, concurrent with downregulation of miR-150. Despite the increasing data from both human and rodent studies, the conserved functional MIAT/miR-150 axis in cardiac pathology has never been investigated in genetically modified mice. Here, we hypothesize that MIAT competitively sequesters miR-150 and blocks the inhibitory effect of miR-150 on proapoptotic genes, thereby increasing maladaptive post-MI remodeling. Using novel mouse models, we demonstrate that global genetic deletion of MIAT in mice protects hearts against MI, while genetic overexpression of MIAT worsens maladaptive cardiac remodeling. Moreover, MIAT downregulates miR-150 in the heart, while miR-150 does not repress MIAT. Importantly, we show that miR-150 overexpression prevents the detrimental post-MI effects caused by overexpression of MIAT (Fig. B). In conclusion, these findings reveal a pivotal functional interaction between MIAT and miR-150 as a novel epigenetic regulatory mechanism pertinent to ischemic cardiac injury.
Abstract 219: NAD Redox Imbalance Accelerates Diabetic Cardiomyopathy via Protein Acetylation and Oxidative Stress
Authors: Chiao Ying Ann , Light Christine , Shi Xiaojian , Tian Rong , Sadoshima Junichi , Gu Haiwei , Lee Chi Fung
Published: October 2020

Diabetes is long linked to lowered NAD/NADH ratio, aka NAD redox imbalance, but its causal role to diabetic cardiomyopathy is not established. We used mouse models with latent decrease in cardiac NAD/NADH ratio (cardiac-specific Ndufs4-KO, cKO) and elevated cardiac NAD levels to directly test whether cardiac NAD redox imbalance accelerates diabetic cardiomyopathy. Control and cKO mice were subjected to 8-week T1D stress, and longitudinal cardiac function was measured by echocardiography. Accelerated declines in systolic and diastolic function were observed in T1D cKO mice. Insulin depletion and hyperglycemia were similar in T1D control and T1D cKO mice, and serum metabolomic analyses showed unchanged aqueous and lipid metabolite levels. These metabolite results suggested that T1D control and cKO hearts were stressed under similar diabetic conditions. Importantly, elevation of cardiac NAD levels to attenuate NAD redox imbalance mitigated the accelerated functional declines in T1D cKO hearts. The data from mouse models with manipulated NAD redox states suggested that NAD redox imbalance accelerates diabetic cardiomyopathy. Cardiac fibrosis levels were not different in T1D control and cKO hearts, while transcript levels of fibrotic genes, including Adamts proteinases, integrins, laminins, matrix metalloproteinases and collagens, also showed no difference. Therefore, the accelerated functional declines in T1D cKO hearts are not due to altered extracellular matrix environment, but are rather due to cardiomyocyte dysfunction. We next determined whether the accelerated cardiac dysfunction is mediated via protein acetylation and oxidative stress. NAD-dependent global protein acetylation and inhibitory acetylation of superoxide dismutase 2 were elevated in T1D cKO hearts. Inhibition of SOD2 concomitantly promoted elevation of protein oxidation levels in T1D cKO hearts. The results suggested that NAD redox balance-dependent protein acetylation regulates oxidative stress to promote diabetic cardiomyopathy.
Abstract 221: Tannic Acid Coated Nanoparticles for Cardiac Regeneration
Authors: Torrieri Giulia , Ferreira Mónica P , Shahbazi Mohammad-Ali , Talman Virpi , Carvalho Cláudia , Karhu Tuuli , Pinto João F , Hirvonen Jouni , Ruskoaho Heikki J , Balasubramanian Vimalkumar , Santos Hélder A
Published: October 2020

The advance of nanomedicines has recently offered novel approaches to tackle cardiovascular diseases and, in particular, myocardial infarction (MI). However, the constant pumping of the heart and the still poor knowledge of targetable moieties, prevented the application of nanomedicines in the cardiovascular field to rise. Tannic acid, a polyphenol derived from plants, has showed affinity for components of the extracellular matrix, in particular elastin, allowing the retention of protein aggregates in the cardiac tissue. Here, we explored the heart targeting abilities of tannic acid by using it to coat spermine modified-acetalated dextran (AcDXSp) nanoparticles (NPs). Briefly, particles were prepared by single emulsion technique and then coated with tannic acid by complexation of the polyphenol with Fe
3+
ions, resulting in the formation of a capsule around the AcDXSp NPs. The nanoparticles were loaded with two small hydrophobic compounds, CHIR99021 and SB431542, which were both proven to increase the efficiency of direct reprogramming of fibroblasts into cardiomyocytes. The biocompatibility of the nanosystem and cellular uptake were performed on both primary rat cardiomyocytes and fibroblasts. The nanoparticles were taken-up by both the cell types and were safe towards primary cardiomyocytes, while the tannic acid coating showed anti-fibrotic effects on primary fibroblasts. Anti-fibrotic effect was further confirmed by RT-qPCR and the effect of the loaded compounds was assessed by β-catenin and Smad3 immunostainings, which demonstrated the ability of the system to induce direct reprogramming of fibroblasts into cardiomyocytes. In particular, the system stabilized β-catenin and prevented the translocation of Smad3 to the nucleus of myo(fibroblasts). In conclusion this nanosystems exhibited potential to tackle the negative fibrosis process occurring after myocardial infarction by both contrasting it, due to the anti-fibrotic effects showed by the tannic acid coating and by potentially regenerating the cardiac tissue, due to the efficient direct reprogramming of fibroblasts into cardiomyocytes exerted by the loaded drugs.
Abstract 232: PDE5A Inhibition in Burn-induced Cardiac Dysfunction
Authors: Wen Jake J , Radhakrishnan Ravi S
Published: October 2020

Severe burn injury initiates a feedback cycle of inflammation, fibrosis and oxidative stress and others that may contribute to heart dysfunction via the PDE5A-cGMP-PKG pathway. Sildenafil (SIL) has been shown decrease immune adverse events in multiple cardiac diseases by inhibition of PDE5A. In this study, using Nano LC MS/MS strategy to identify differentially expressed proteins, we will determined the pathological importance of PDE5A-dependent injury in burn-related heart dysfunction using our well-established rat model with 4 groups: sham; sham/SIL (sham with sildenafil injection); 24 hpb (60% total body surface area scald burn and harvested at 24 hours post burn) and 24 hpb/SIL (24 hpb with SIL immediately after burn). We performed proteomics, Vevo 2100 ECG, qPCR, histology and ELISA to test our hypothesis. Our results show Sildenafil 1) inhibited burn-induced PDE5A mRNA, 2) increased cGMP and PKG activity leading to recover of PKG down-regulated genes (IRAG, PLB, RGS2, RhoA and MYTP), 3) decreased the ROS level (H2O2), oxidatively modified adducts (malonyldialdehyde (MDA), carbonyls), fibrotic gene expression (ANP, BNP, COL1A2, COL3A2, aSMA and αsk-Actin), inflammation-related gene expression (RELA, IL-18 and TGF-β) from burned rats (BRs) and 4) preserved Left-ventricular heart function (CO, EF, SV, LVvol at systolic, LVPW at diastolic and FS) and restored the oxidant/antioxidant balance (total antioxidant, total SOD activity and Cu,ZnSOD activity) in BRs. We demonstrate that the PDE5A-cGMP-PKG pathway mediates burn-induced heart dysfunction and will provide the theoretic foundation to guide pre-clinical translational study of Sildenafil in burned patients.
Abstract 235: Hypertrophic Cardiomyopathy, a Disease of Altered Cardiac Energetics
Authors: Ranjbarvaziri Sara , Ellenberger Mathew , Kooiker Kristi , Fajardo Giovanni , Zhao Mingming , Schroer Alison , Woo Y Joseph Y , Ruppel Kathleen M , Spudich James A , Snyder Michael , Contrepois Kevin , Bernstein Daniel
Published: October 2020

Hypertrophic cardiomyopathy (HCM) is a complex disease, the phenotypes of which are only partly explained by the biomechanical effects of individual genetic variants. At the cellular level, HCM sarcomeric mutations generally enhance maximal force production ultimately leading to higher energy demands. Despite significant advances in elucidating sarcomeric structure-function relationships, there is limited knowledge on the link between altered cardiac energetics and HCM phenotypes. In this work, we test the hypothesis that changes in cardiac energetics are a common pathway leading to the clinical pathophysiological phenotypes of HCM.

We performed a comprehensive multi-omic study of the molecular, ultrastructural, and functional features of HCM energetics using septal myectomy samples from 27 HCM patients and 13 normal controls (donor hearts). Combined mass spectrometry and RNA-Seq revealed dramatic alterations in multiple metabolic pathways,with major dysregulation in fatty acid metabolism leading to reduced acylcarnitines and accumulation of free fatty acids. Additionally, HCM hearts showed clear signs of global energetic decompensation manifested by a decrease in high energy phosphate metabolites (ATP, ADP, and PCr) and reduction in several mitochondrial genes involved in creatine kinase and ATP synthesis machinery. Quantitative electron microscopy showed a marked increase in severely damaged mitochondria with reduced cristae density, affecting 10-12% of total mitochondria, coinciding with reduced citrate synthase (CS) activity and mitochondrial respiration. These mitochondrial abnormalities were associated with elevated ROS and reduced antioxidant defenses along with insufficient mitophagic clearance.

Overall, our findings suggest that perturbed metabolic signaling and mitochondrial dysfunction are common pathogenic mechanisms in HCM. A central role for compromised energetics in HCM could also help explain the delayed age of onset of the clinical phenotype and present novel drug targets for attenuation of the clinical disease through reducing mitochondrial injury and improving function.
Abstract 238: Single-Cell Transcriptomic Analysis and Patient-Specific IPSCs Reveal Dysregulated Cell Cycle in Coronary Endothelial Cell in Hypoplastic Left Heart Syndrome
Authors: Gu Mingxia , Miao Yifei , Zhou Xin , Tian Lei , Martin Marcy , Nelson Timothy J
Published: October 2020

Hypoplastic left heart syndrome (HLHS) is a single ventricle congenital heart disease that results in severe underdevelopment of the left ventricle, mitral valve, aortic valve, and ascending aorta. Early serial postmortem examinations also revealed a high rate of coronary anomalies in HLHS, which included multiple ventriculo-coronary arterial connections as well as thick-walled and kinked coronary arteries. A previous study showed that fetal hypoplastic left hearts had a reduced endothelial cell (EC) population and lower capillary density compared with normal hearts. However, the mechanism underlying coronary abnormalities associated with HLHS remains unknown. Thus, we generated induced pluripotent stem cells derived ECs (iPSC-ECs) from three HLHS patients and three age-matched controls. Single Cell RNA-Seq (scRNA-seq) profiling identified both endocardial (NPR3
+
/CDH5
+
) and coronary endothelial populations (APLN
+
/CDH5
+
) from the heterogeneous iPSC-ECs. Intriguingly, a subcluster of the coronary endothelial cells (CECs) with cell cycle arrest was specifically enriched in HLHS patients. Further cell cycle analysis showed that 30.6% of the HLHS cells were trapped in the G1 phase, while the majority of the control CECs entered cell cycle normally. Additionally, the cell cycle differences between control and HLHS was only seen in CECs, not in the endocardial population. To verify our transcriptomic analysis, we applied negative cell sorting (NPR3
-
/CDH5
+
) on iPSC-ECs to purify CECs (iCECs) and confirmed that HLHS iCECs showed profound reduction of cell cycle/proliferative genes (
KI67, PCNA, CCNA2, CCNB1
) and abnormal induction of
CCND2
, which is the hallmark of G1 phase. BrdU assays also indicated suppressed proliferation in HLHS iCECs. Furthermore, we profiled the transcriptome from a human heart with an underdevelopment left ventricle (ULV) at single cell resolution. When compared to the normal human heart, pathway enrichment analysis of differentially expressed genes in ULV hearts demonstrated reduced cell proliferation in the CEC subpopulation. Here, we identified that CECs from HLHS patients exerted proliferative defects that can potentially impede the development of vascular/capillary structure and cause related functional deficiencies. Reformation of the coronary defect provides a promising therapeutic strategy to prevent HLHS deterioration.
Abstract 243: Myocardial-associated B Cells Modulate the Composition of Cardiac Resident Macrophages
Authors: Rocha Resende Cibele , Adamo Luigi
Published: October 2020

Background:
The murine myocardium contains distinct subsets of macrophages with disparate roles that can be defined by expression levels of CCR2 and MHC-II. The factors that modulate the composition of the myocardial macrophage pool in the naïve heart are not completely known. B cells have been recently shown to modulate the prevalence of T cells in the heart but the relationship between B cells and myocardial macrophage subsets remains unknown.

Objective:
To test the hypothesis that myocardial-associated B cells modulate the composition of the myocardial macrophage pool.

Methods and Results:
Hearts from B cell deficient (μMT) or B cell depleted (treated with Anti-CD20 antibody) mice and age/sex matched controls were analyzed via flow cytometry at post-natal day 14 and soon after weaning. Absence of B cells was consistently associated with a decrease in the prevalence of CCR2
-
MHC-II
high
resident macrophages. Immunofluorescence analysis of the heart showed that the myocardial-associated B cells were frequently in close proximity of CD68+ myocardial macrophages. In vitro co-culture of myocardial-derived CD64+ macrophages with B cells or with B cell-conditioned media showed that B cells fostered an increase in the prevalence of CCR2
-
MHC-II
high
macrophages through a direct paracrine effect. In vivo 10X single cells sequencing and flow cytometric analysis together with in vitro ELISA and antibody mediate blocking suggested that the paracrine effect of B cells on myocardial macrophages is mediated, at least in part, by IL-16 secretion.

Conclusion:
Myocardial-associated B cells modulate the phenotype of myocardial resident macrophages. This is likely the result of a direct paracrine effect mediated, at least in part, by IL-16. These findings describe for the first time a relationship between circulating naïve B cells and tissue resident macrophages and unveil a previously unappreciated relationship between lymphoid and myeloid myocardial cells.
Abstract 254: Profiling Transcriptional Changes and Identifying Novel Molecular Markers During the Dedifferentiation of Cardiomyocyte
Authors: Zhu Yike , Tan Lek Wen Wilson , Dang Vinh Do , Ackers-Johnson Matthew A , Foo Roger
Published: October 2020

The loss of cardiomyocytes (CMs) during heart failure (HF) cannot be replaced by new CMs due to their limited proliferative capacity. Regenerating the failing heart by promoting CM cell-cycle re-entry has thus become a possible solution rigorously pursued. Some genes have been proven to promote endogenous CM proliferation, nearly always preceded by CM dedifferentiation, wherein terminally-differentiated CMs are reversed back initially to a less-matured status, preceding cell division. However, very little else is known about CM dedifferentiation, including the lack of robust molecular markers, the mechanisms driving it, whether it necessarily leads to CM proliferation, and the effect of CM dedifferentiation alone on cardiac function. Therefore, investigating CM dedifferentiation is of great significance. Here, we profiled genome-wide transcriptional changes and identified novel molecular markers during CM dedifferentiation. We used the established
in vitro
model where adult mouse CMs dedifferentiated and re-differentiated (DR) upon 2-week long culture, without detectable cell division. DR was evident by breakdown and rebuilding of sarcomeres, and differential regulation of cardiac fetal genes. Tracking the transcriptome change pointed to gene pathways involved in this process and potential markers of CM dedifferentiation. Another assumption is that transient induction of
Yamanaka
factors (
Oct4, Sox2, Klf4, Myc, OSKM
) induces CM dedifferentiation of CMs
in vivo
, since the four factors are themselves sufficient to induce pluripotency of human and mouse somatic cells. Our preliminary results show that CM-specific overexpression of
OSKM
using adeno-associated virus serotype 9 (AAV9) induced fetal-like hallmarks, down-regulation of sarcomeric genes, re-activation of cell cycle, reduced fatty acid utilization, and associated loss of cardiac function . Putative markers identified in the
in vitro
model was also validated to be upregulated in
OSKM
-overexpressed CMs. In conclusion, through 2 models of CM dedifferentiation, we have screened out potential novel markers and regulators of CM dedifferentiation which now requires further validation.
Abstract 262: Regional Transcriptomics of Left Atrial Cardiac Tissue in a Patient With Atrial Fibrillation
Authors: Harwood Samuel J , Barnard John , Sun Han , McCurry Kenneth R , Okamoto Toshihiro , SMITH Jonathan D , Van Wagoner David R , Chung Mina K
Published: October 2020

Background:
Common genetic variants and inflammatory factors are associated with atrial fibrillation (AF) risk. Although the top AF risk locus is near
PITX2
(associated with pulmonary vein (PV) development), regional differential expression (DE) of AF risk genes in the left atrium (LA) and PVs is not well studied. While LAA and PV gene expression have been compared, transcriptome mapping of the entire LA has not been done. We tested the hypothesis that there is significant regional DE in LA structures.

Methods:
RNAseq was performed in 25 regions within the PVs (n=12), LA body (LAB) (n=10), and LA appendage (LAA) (n=3) from a 75 year old male with hypertension and AF who died of a stroke. DE analysis of regional clusters (R 3.6
Limma
package) and gene set enrichment analysis (GSEA) (Broad Institute) were performed.

Results:
In full transcriptome (n=20664) sequencing, 118 genes were significantly (q < 0.05) upregulated in the LAA, including
FOSB
(fold change [FC] 67.0),
IL6
(FC 15.4),
TNF
(FC 2.4) and
NLRP3
(FC 2.4), and 56 downregulated, including developmental genes such as
SHOX2
(FC -12.9) and
HOXA4
(FC -4.1). LAA GSEA showed enrichment (FDR < 0.05) of inflammatory response genes and TNF-α, IL-2, IL-6, and IgG signaling pathways. 2454 genes were significantly upregulated and 3737 downregulated in the PVs while 4021 genes were upregulated and 2998 were downregulated in the LAB. In an AF implicated gene set (n=190), 28 genes were upregulated in the PVs, including
SIRT1
(FC 1.7), and 31 downregulated, including
PMNT
(FC -2.1) and
CGA
(FC -3.2).
NEURL1
was significantly upregulated in the inferior PV (n=4) compared to the superior PV (n=8) (FC 3.6). In the LAB, 33 genes were upregulated, including
MYH7
(FC 2.2)
and PMNT
(FC 2.1)
,
and 38 downregulated including
SIRT1
(FC -1.9).
PITX2
did not show any significant DE in any gene set.

Conclusions:
This data shows that genes involved in AF pathogenesis can have substantial regional expression difference, particularly when comparing the LA body, PVs, and LAA. Inflammatory activation in the LAA may contribute to increased stroke risk.
Abstract 263: Loss of the RNA-binding Protein ZFP36L2 Results in Peri-partum Cardiomyopathy Through Dysregulation of the P53-mTOR Pathway
Authors: Tatekoshi Yuki , Kouzu Hidemichi , Shapiro Jason S , Chang Hsiang-Chun , Jesus Adam D , Stanczyk Paulina J , Ben-Sahra Issam , Blackshear Perry , Ardehali Hossein
Published: October 2020

Introduction:
A loss of function mutation in the mRNA binding protein ZFP36L2 has been associated with congenital heart diseases in humans, although the mechanism remains unclear.

Objectives:
We sought to elucidate the role of ZFP36L2 in the heart using mice with cardiac-specific ZFP36L2 deletion.

Results:
Cardiac-specific
Zfp36l2
knockout (cs-L2KO) female mice exhibit a dramatic phenotype in which the majority of mice die after repeated pregnancies due to heart failure. We performed RNA-seq in H9c2 cells with ZFP36L2 KD, and found the expression of multiple mTOR pathway genes were altered, including marked repression of
Redd1
and
Sesn2
. Consistent with the RNA-seq data, the activity of mTORC1 was increased in cs-L2KO mouse hearts and in ZFP36L2 KD H9c2 cells. Loss of TSC2 mitigated mTORC1 hyperactivity in ZFP36L2 KD cells, indicating that ZFP36L2 regulates mTORC1 activity through the TSC2 complex. Overexpression of REDD1 rescued increased mTORC1 activity with ZFP36L2 KD, suggesting that ZFP36L2 regulates mTORC1, in-part through REDD1. Under total amino acid starvation, we observed higher p-S6K
T389
levels in ZFP36L2 KD cells, and SESN2 overexpression mitigated p-S6K
T389
, indicating loss of ZFP36L2 also regulates mTORC1 through amino acid sensing. p53 is a transcriptional activator of both REDD1 and SESN2, and we found that ZFP36L2 was required to maintain p53 levels by directly destabilizing MDM2 mRNA. Accordingly, administration of Nutlin3, an MDM2 inhibitor, reversed the reduction in p53, REDD1 and SESN2 protein levels, and prevented the increase in S6K phosphorylation in response to ZFP36L2 KD. Next, we treated pregnant mice with rapamycin, and cs-L2KO mice treated with rapamycin displayed significantly improved cardiac function after delivery. Finally, we analyzed human heart samples from patients with peri-partum cardiomyopathy and found ZFP36L2 protein to be significantly decreased and p-S6K
T389
levels increased, compared to healthy controls.

Conclusions:
Our studies demonstrate that ZFP36L2 regulates mTORC1 activity through modifying p53 stability, and that the disruption of this pathway induces peri-partum cardiomyopathy. Inhibition of mTORC1 hyperactivity with rapamycin can be a potential therapy for this disease.