bluuebunny/crossref_metadata_2025 huggingface dataset:
A Comparative Study of Impeller Modification Techniques on the Performance of the Pump as a Turbine
Authors: Nasir Abdulbasit , Dribssa Edessa , Girma Misrak , Demeke Tamerat
Published: November 2022

The extensive use of the pump as a turbine (PAT) for micro-hydropower applications has a significant value from economic and technical viewpoints. However, the unavailability of the characteristics curve and relatively lower efficiency are the two basic limitations when considering pumps for power-generating applications. In this paper, the performance of the PAT is analyzed using the computational fluid dynamics (CFD) software called Ansys CFX in conjunction with standard

k

-

ε

. Then, experiments were done to verify the results of the simulation. Measurement inaccuracy effects are also taken into account. The initial performance of the PAT is refined by controlling basic design parameters (i.e., increasing the number of impeller blades, decreasing blade thickness, blade tip rounding, and adjusting blade inlet angle). Additionally, a new modification method known as blade grooving is also introduced in this research. Finally, all listed modification techniques are applied simultaneously to achieve maximum performance. The output of the study confirms that the adopted modification techniques have a positive effect on performance improvement. When the number of impellers is increased, the power output is enhanced by 5.72%, and blade grooving provides the most efficiency improvement, i.e., 7.00%. But decreasing blade thickness has no remarkable impact on the performance; the power output and efficiency are improved by 1.24% and 2.60%, respectively. The maximum performance improvement was achieved when the modification techniques are applied simultaneously with 10.56 and 10.20 percent of power and efficiency increments, respectively. From the entire study, it can be concluded that the chosen design parameters have an important effect on stabilizing the internal flow, decreasing the required head, decreasing the hydraulic loss in the impeller, and increasing the overall performance. The study also helps to figure out which modification technique is the most practical.
Approximate Biprojectivity of ℓ1‐Munn Banach Algebras
Authors: Zarei G. , Pourabbas A. , Rostami M. , Sahami A.
Published: November 2022

In the present paper, we study the approximate biprojectivity and weak approximate biprojectivity of ℓ1‐Munn Banach algebras when the related sandwich matrix is regular over Inv(A). In fact, we show that a ℓ1‐Munn Banach algebra with the regular sandwich matrix over Inv(A) is approximately biprojective (weak approximately biprojective) if and only if A is approximately biprojective (weak approximately biprojective), respectively. We also study approximate biprojectivity of upper triangular Banach algebra when the associated sandwich matrix with elements in Inv(A) is invertible. Finally, we apply our results to Rees semigroup algebras.
Microbiota and Gut Health: Promising Prospects for Clinical Trials from Bench to Bedside
Authors: Wang Jie , Zhang Puxuan , Chen Shujie , Duan Huimin , Xie Liwei
Published: November 2022

Inflammatory bowel disease (IBD) is a chronic inflammatory gastrointestinal disease characterized by weight loss, abdominal pain, and bloody diarrhea. The number of affected patients has increased in recent years. Despite the fact that scientists have been studying the pathogenesis of IBD for many years, the specific pathogenesis pathway remains unclear. As a result, none of the therapeutic approaches can cure IBD patients completely. However, the increasing research factors associated with the incidence of IBD are reasonable. These variables can be divided into two categories: microbiome-related factors (bacteria, fungi, and viruses) and nonmicrobiome-related factors (diet, gene, host immune system, gender, and ethnicity). Surprisingly, we found that all the variables impact the gut flora in IBD patients, either directly or indirectly. Dysbiosis of the gut microbiota eventually leads to an increase in the incidence of IBD. As a result, therapeutic targets focusing on correcting dysbiosis in the gut microbiome, including using probiotics and postbiotics, could become one of the most promising IBD treatments in the future. We went through each linked factor and explained how they contribute to an increased risk of IBD. We will review some existing conventional therapies for IBD before moving on to a revolutionary therapy strategy that employs prebiotics, probiotics, and postbiotics to treat IBD based on the criteria stated. Furthermore, different persons have varying reactions to the same probiotic strain. As a result, we also provide the option of having individualized probiotic medication tailored to each IBD patient.
Deep Learning in Ischemic Stroke Imaging Analysis: A Comprehensive Review
Authors: Cui Liyuan , Fan Zhiyuan , Yang Yingjian , Liu Rui , Wang Dajiang , Feng Yingying , Lu Jiahui , Fan Yifeng
Published: November 2022

Ischemic stroke is a cerebrovascular disease with a high morbidity and mortality rate, which poses a serious challenge to human health and life. Meanwhile, the management of ischemic stroke remains highly dependent on manual visual analysis of noncontrast computed tomography (CT) or magnetic resonance imaging (MRI). However, artifacts and noise of the equipment as well as the radiologist experience play a significant role on diagnostic accuracy. To overcome these defects, the number of computer‐aided diagnostic (CAD) methods for ischemic stroke is increasing substantially during the past decade. Particularly, deep learning models with massive data learning capabilities are recognized as powerful auxiliary tools for the acute intervention and guiding prognosis of ischemic stroke. To select appropriate interventions, facilitate clinical practice, and improve the clinical outcomes of patients, this review firstly surveys the current state‐of‐the‐art deep learning technology. Then, we summarized the major applications in acute ischemic stroke imaging, particularly in exploring the potential function of stroke diagnosis and multimodal prognostication. Finally, we sketched out the current problems and prospects.
Effect of Analytical Particle Size on Pore Structure of High Volatile Bituminous Coal and Anthracite Using Low-Pressure N2 and CO2 Adsorption
Authors: Dang Zheng , Wang Xiaoming , Hou Shihui , Pan Sidong
Published: November 2022

To investigate the effect of analytical particle size on pore structure, mesopore (2-50 nm) and micropore (<2 nm) characteristics of high volatile bituminous coal and anthracite with different particle size were determined using low-pressure N2/CO2 adsorption analyses. Mesopore structure parameters in the two coals increase with decreasing particle size, which are attributed to the opening of closed mesopores during the pulverization process. The closed mesopores with different pore size ranges are opened with a certain percentage in high volatile bituminous coal, but opened irregularly in anthracite during pulverization. Micropore structure parameters of the two coals show different variations with decreasing particle size, which are not related to the reconstituted micropore structure. Mineral matter contributes more mesopores than organic matter in anthracite and exerts the negative effect on micropore in the two coals. An evolution model is established to elaborately describe the change of pore structure during the pulverization process, where mineral matter plays a mediating role in the effect of particle size on pore structure.
Q‐Differential Equations of Higher Order and Structural Properties of the Approximate Roots for q‐Modified Derangements’ Polynomials
Authors: Ryoo Cheon Seoung , Kang Jung Yoog
Published: November 2022

The objective of this study is to find q‐differential equations of higher order related to q‐modified derangements’ polynomials and confirm the structure of approximation roots. Furthermore, it states several symmetric properties of q‐differential equations of higher order and the special properties of the approximation roots of q‐modified derangements’ polynomials.
A New Hybrid Conjugate Gradient Projection Method for Solving Nonlinear Monotone Equations
Authors: Fang Minglei , Wang Min , Ding Defeng
Published: November 2022

In this study, we propose a new modified hybrid conjugate gradient projection method with a new scale parameter φk for solving large‐scale nonlinear monotone equations. The proposed method includes two major features: projection techniques and sufficient descent property independent of line search technique. Global convergence of the proposed method is proved under some suitable assumptions. Finally, numerical results illustrating the robustness of the suggested strategy and its comparisons are shown.
Analysis of the Pile-Soil Combined Effect of Micropiles in Strengthening Expansive Soil Landslide in Southern Shaanxi Based on the Dynamic Soil Arch Model
Authors: Ding Xiao , He Hui , Qu Yonglong , Yang Yixuan
Published: November 2022

Southern Shaanxi region is mountainous and landslide disasters are frequent. Micropile reinforcement technology is widely used in landslide management, especially in landslide emergency rescue projects, due to easy construction, small soil disturbance, and strong site adaptability. Soil arch effect is an important prerequisite for safe and economic performance of the support function of micropile. For the swelling soil landslide of the accumulation layer in southern Shaanxi, firstly, a numerical model is established by means of thermal-mechanical coupling of FLAC3D to explore the variation of the soil arch form with different swelling forces. Then, two theoretical models are proposed under normal working condition and rainfall working condition. The evolution and failure mode of soil arch are analyzed. The results show that the soil expansion after rainfall causes the change of the soil arch mode, the range of pile-soil interaction is obviously reduced, and the soil arch effect is weakened. It should be considered in the design of micropiles in strengthening expansive soil landslide.
Human Metastatic Cholangiocarcinoma Patient-Derived Xenografts and Tumoroids for Preclinical Drug Evaluation
Authors: Serra-Camprubí Queralt , Verdaguer Helena , Oliveros Winona , Lupión-Garcia Núria , Llop-Guevara Alba , Molina Cristina , Vila-Casadesús Maria , Turpin Anthony , Neuzillet Cindy , Frigola Joan , Querol Jessica , Yáñez-Bartolomé Mariana , Castet Florian , Fabregat-Franco Carles , Escudero-Iriarte Carmen , Escorihuela Marta , Arenas Enrique J. , Bernadó-Morales Cristina , Haro Noemí , Giles Francis J. , Pozo Óscar J. , Miquel Josep M. , Nuciforo Paolo G. , Vivancos Ana , Melé Marta , Serra Violeta , Arribas Joaquín , Tabernero Josep , Peiró Sandra , Macarulla Teresa , Tian Tian V.
Published: November 2022

Purpose:

Cholangiocarcinoma (CCA) is usually diagnosed at advanced stages, with limited therapeutic options. Preclinical models focused on unresectable metastatic CCA are necessary to develop rational treatments. Pathogenic mutations in IDH1/2, ARID1A/B, BAP1, and BRCA1/2 have been identified in 30%–50% of patients with CCA. Several types of tumor cells harboring these mutations exhibit homologous recombination deficiency (HRD) phenotype with enhanced sensitivity to PARP inhibitors (PARPi). However, PARPi treatment has not yet been tested for effectiveness in patient-derived models of advanced CCA.

Experimental Design:

We have established a collection of patient-derived xenografts from patients with unresectable metastatic CCA (CCA_PDX). The CCA_PDXs were characterized at both histopathologic and genomic levels. We optimized a protocol to generate CCA tumoroids from CCA_PDXs. We tested the effects of PARPis in both CCA tumoroids and CCA_PDXs. Finally, we used the RAD51 assay to evaluate the HRD status of CCA tissues.

Results:

This collection of CCA_PDXs recapitulates the histopathologic and molecular features of their original tumors. PARPi treatments inhibited the growth of CCA tumoroids and CCA_PDXs with pathogenic mutations of BRCA2, but not those with mutations of IDH1, ARID1A, or BAP1. In line with these findings, only CCA_PDX and CCA patient biopsy samples with mutations of BRCA2 showed RAD51 scores compatible with HRD.

Conclusions:

Our results suggest that patients with advanced CCA with pathogenic mutations of BRCA2, but not those with mutations of IDH1, ARID1A, or BAP1, are likely to benefit from PARPi therapy. This collection of CCA_PDXs provides new opportunities for evaluating drug response and prioritizing clinical trials.
GSK3β Inhibition Prevents Macrophage Reprogramming by High-Dose Methotrexate
Authors: Ríos Israel , López-Navarro Baltasar , Torres-Torresano Mónica , Soler Palacios Blanca , Simón-Fuentes Miriam , Domínguez-Soto Ángeles , Muller Ittai B. , Jansen Gerrit , Corbí Ángel L. , Puig-Kröger Amaya
Published: November 2022

Methotrexate (MTX) is an antifolate drug used as a chemotherapeutic agent for acute lymphoblastic leukemia, where MTX improves patients’ prognosis. Macrophage reprogramming is being increasingly assessed as an antitumor therapeutic strategy. However, and although MTX limits the pathogenic action of macrophages in chronic inflammatory diseases, its effects on tumor-promoting macrophages have not been previously explored. We now report that MTX shapes the transcriptional and functional profile of M-CSF-dependent human macrophages, whose transcriptome is highly enriched in the gene signature that defines pathogenic tumor-associated macrophages (“large TAM”). Specifically, MTX prompted the acquisition of the gene signature of antitumoral “small TAM” and skewed macrophages toward an IL-6high IFNβ1high IL-10low phenotype upon subsequent stimulation. Mechanistically, the MTX-induced macrophage reprogramming effect correlated with a reduction of the M-CSF receptor CSF1R expression and function, as well as a diminished expression of MAF and MAFB transcription factors, primary determinants of pro-tumoral macrophages whose transcriptional activity is dependent on GSK3β. Indeed, the ability of MTX to transcriptionally reprogram macrophages toward an antitumoral phenotype was abrogated by inhibition of GSK3β. Globally, our results establish MTX as a macrophage reprogramming drug and indicate that its ability to modulate macrophage polarization may also underlie its therapeutic benefits. Since GSK3β inhibition abrogates the reprogramming action of MTX, our results suggest that the GSK3β-MAFB/MAF axis constitutes a target for the macrophage-centered antitumor strategies.
The Mechanism of Delayed Ischemic Preconditioning in Alleviating Acute Ischemia/Reperfusion Renal Injury through Treg Mediated by Immature CD11c+ Dendritic Cells
Authors: Yang Wenjuan , Peng Tao , Shi Chunli , Cui Fang , Chen Menghua , Zhang Ting
Published: November 2022

Introduction: Renal ischemia-reperfusion injury (IRI) is one of the major causes of acute kidney injury, and its mechanism is complex involving multiple factors, while delayed ischemic preconditioning (DIPC) has a protective effect on the above process. In our previous study, we found that DIPC can exert its protection on renal IRI by inhibiting the maturation of dendritic cells (DCs), but the mechanism has not been clarified. This study aimed to investigate the protective mechanism of DIPC on renal IRI in mice through Treg mediated by immature DCs (imDCs). Methods: The IRI mice model, DIPC treatment, and conditional CD11c+ DCs (CD11c-DTR) knockout mice were used to perform our study. The maturation and differentiation of DCs and Treg cells in the kidney and spleen were analyzed by flow cytometry. HE staining was used to evaluate the pathology of the kidney tissue. The level of creatinine (Cr), oxidative stress factors (SOD, MDA), and inflammatory factors (TNF-α, IL-10, IL-4) were also measured. Then, imDCs were co-cultured with HK-2 cells, and apoptosis was analyzed with flow cytometry and PI-Hoechst 33,342 fluorescence staining to assess the apoptosis rate of HK-2 cells under hypoxic-reoxygenated (H/R) conditions. Results: DIPC could decrease renal Cr levels, alleviate pathological renal damage, and reduce oxidative stress and inflammation caused by IRI. Moreover, DIPC could decrease the number of mature DCs (mDCs) and increase Treg lymphocyte infiltration in the kidney tissue, while the reduction of DCs reversed this process. In addition, our in vitro experiment found that in the H/R model, the apoptosis of HK-2 cells decreased which were co-cultured with imDCs. Conclusion: DIPC can regulate the differentiation of DCs into imDCs, thus affecting the differentiation level and distribution of Treg cells to exert its protective effect on renal IRI.
Preoperative Predictive Nomogram Based on Alanine Aminotransferase, Prothrombin Time Activity, and Remnant Liver Proportion (APART Score) to Predict Post-Hepatectomy Liver Failure after Major Hepatectomy
Authors: Maehira Hiromitsu , Iida Hiroya , Mori Haruki , Nitta Nobuhito , Maekawa Takeru , Takebayashi Katsushi , Kaida Sachiko , Miyake Toru , Tani Masaji
Published: November 2022

Introduction: Post-hepatectomy liver failure (PHLF) is a serious complication associated with major hepatectomies. An accurate prediction of PHLF is necessary to determine the feasibility of major hepatectomy. This study aimed to assess the association between PHLF and preoperative laboratory and computed tomography (CT) findings. Methods: Medical records of 65 patients who underwent major hepatectomy and preoperative CT were retrospectively reviewed. We evaluated future remnant liver volume evaluation models and remnant liver hemodynamics, which were assessed by arterial enhancement fraction (AEF) by using preoperative CT. Variables, including CT findings, were compared between patients with and without PHLF after major hepatectomy, and the preoperative PHLF-predicting nomogram was constructed using multivariate logistic regression. Results: The PHLF group included 21 patients (32.3%). The AEF was not significantly different between the two groups. In the future remnant liver volume evaluation models, future remnant liver proportion (fRLP) had the highest concordance index (C-index) in the receiver operating characteristic curve analysis (C-index, 0.755). Multivariate analysis of preoperative evaluable factors revealed that alanine aminotransferase levels (p = 0.034), prothrombin time activity (p = 0.021), and fRLP (p = 0.012) were independent predictive factors of PHLF. A nomogram (APART score) was constructed using these three factors, with a receiver operating curve showing a C-index of 0.894. According to the APART score, scores of 51–60 indicated moderate risk (40.0%), and scores over 60 indicated a high risk of PHLF (83.3%) (p < 0.001). Discussion: The APART score may help predict PHLF in patients indicated for major hepatectomies.
Abstract EC105: A Mouse Model Carrying The R502W Mutation In Cardiac Myosin Binding Protein C (cMyBP-C) Leads To Hypertrophic Myocardial Remodeling In The Absence Of  Haploinsufficiency
Authors: Sen-Martín Laura , Fernández-Trasancos Ángel , Lopez-Unzu Miguel , Sánchez-Ortiz David , Pricolo María Rosaria , Alegre-Cebollada Jorge
Published: November 2022

Background:
Familial cardiomyopathies are caused by inherited genetic mutations resulting in anatomical and/or mechanical alterations that compromise cardiac function. The missense variant R502W in cardiac myosin-binding protein C (cMyBP-C) is the most frequent mutation leading to hypertrophy cardiomyopathy (HCM) in humans. Protein and mRNA levels have been proposed not to be altered by this mutation, which points towards pathomechanisms not involving cMyBP-C haploinsufficiency. To test the hypothesis that R502W variant induces HCM remodeling despite preserved cMyBP-C levels, CRISPR/Cas9-based genetic engineering was used to generate a knock-in (KI) mouse model harboring the R502W mutation in murine cMyBP-C.

Methods:
Histology, echocardiography and magnetic resonance imaging were employed to characterize the cardiac phenotype of KI mice. Protein and cMyBP-C mRNA levels were quantified by Western Blot and qPCR. Immunofuorescence in neonatal cardiomyocytes was used to analyze protein localization. Alteration of myosin conformations was assessed in permeabilized myofibrils using Mant-ATP pulse-chase experiments. Transcriptomic analysis was performed to confirm upregulation of hypertrophic cardiac remodeling programs.

Results:
We observe higher heart weight to tibia length ratio starting at 18 weeks old homozygous KI mice, which is accompanied by thicker trabeculae, altered ventricle geometry, reduced left ventricular systolic and diastolic function. Fibrosis and hypertrophy at the cardiomyocyte level were detected at 18 weeks of age. Biochemical and immunofluorescence analyses demonstrate that cMyBP-C mRNA, protein levels and localization are not altered by the mutation. Myosin conformations appear largely unaffected in homozygous KI mice.

Conclusion:
We propose that pathogenicity of the R502W mutation stems from mechanisms that do not involve cMyBP-C haploinsufficiency. We expect that further investigation of R502W mice will shed light on the molecular triggers of HCM caused by cMyBP-C point mutations not triggering protein haploinsufficiency.
Abstract GS107: Nil10: A Newly Synthesized Nanoparticle That Reduces Inflammation And Preserves Heart Function In Murine And Porcine Models Of Acute Myocardial Infarction
Authors: Tesoro Laura , Botana Laura , Hernandez Navarro Ignacio , Ramirez-Carracedo Rafael , Zamorano Jose L , Saura Marta , Zaragoza Carlos
Published: November 2022

Introduction:
Interleukin-10 (IL-10) has long been recognized as a key anti-inflammatory cytokine, whose expression is associated to macrophage polarization towards resolution after acute myocardial infarction.

Hypothesis:
Targeting the IL10 receptor (IL10R) in patients undergoing AMI, may represent a new strategy to improve cardiac function.

Methods and Results:
We synthesized NIL10, a micelle-based nanoparticle conjugated to an IL-10 agonist, to analyze the effect on cardiac function and inflammation resolution after cardiac ischemia/reperfusion. After confirmation of NIL10 (conjugated with Rhodamine) co-localization to IL-10R, as detected by immunohistofluorescence (Fig. 1A), intravenous administration of 1 mg/kg NIL10 in mice and pigs subjected to AMI, efficiently decreased left ventricle fibrosis and significantly improved left ventricle ejection fraction (LVEF), by day 7 after injection, when compared to animals injected with, NIL10SC, a nanoparticle control (Fig. 1B-C). As expected, NIL10 induced macrophage polarization towards M2-like macrophages after day 3 of reperfusion (Fig D), by at least promoting IL-10R activity, as shown by increased phosphorylation of IL-10R and the downstream STAT3 in the necrotic area of the heart (Figure 1E). STAT3-dependent inflammation inhibition was assayed by administration of 50μM NIL10 in RAW-247 cells stimulated with 500 μM LPS, in which nuclear translocation of NF-κB was significantly reduced (Fig. 1E) by at least phosphorylation of I-κB-α.

Conclusions:
Our findings show NIL10 as a new tool to improve cardiac function in hearts under AMI, at least through inhibition of STAT3-induced nuclear translocation of NF-κB.
Abstract GS108: Troponin I Tyrosine-26 Phosphorylation Improves Relaxation And Is Beneficial During Pathological Diastolic Dysfunction
Authors: Salyer Lorien , Sturgill Sarah , Brundage Elizabeth , Salhi Hussam , Shettigar Vikram , Abay Eaman , Ziolo Mark T , Biesiadecki Brandon
Published: November 2022

Diastolic dysfunction contributes to disease in both heart failure with preserved and reduced ejection fraction. There are currently no approved therapies to accelerate impaired relaxation for heart failure patients with diastolic dysfunction. A healthy heart can modify its ability to relax through post-translational modifications (e.g., phosphorylation) of myofilament proteins. The inhibitory subunit of the troponin complex, troponin I (TnI), is a key regulator of cardiac contraction and relaxation and is endogenously phosphorylated at Tyr-26. We previously demonstrated that this novel tyrosine phosphorylation decreases calcium sensitivity and accelerates calcium dissociation from troponin C. We therefore hypothesize that increasing TnI Tyr-26 phosphorylation will accelerate relaxation and be beneficial during pathological diastolic dysfunction. To determine the effects of TnI Tyr-26 phosphorylation
in vivo
, we generated a phospho-mimetic mouse with TnI Tyr-26 mutated to Glu. Structural and functional echocardiography and hemodynamics measurements demonstrate that TnI Tyr-26 phosphorylation accelerates relaxation and increases diastolic function
in vivo
. To determine the effect of this TnI Tyr-26 phosphorylation mediated acceleration of diastolic function during disease, we performed unilateral nephrectomy with DOCA-salt (neph/DOCA) to induce diastolic dysfunction. Wild-type mice subjected to neph/DOCA develop left ventricular hypertrophy, enlarged left atria, elevated diastolic pressure, and slowed relaxation, indicative of diastolic dysfunction. In contrast with wild-type, TnI Tyr-26 phosphorylation mice subjected to neph/DOCA display normal diastolic function without any cardiac remodeling. Overall, we demonstrate that TnI Tyr-26 phosphorylation inhibits the development of diastolic dysfunction and is therefore a novel mechanism to accelerate myocardial relaxation
in vivo.
Abstract GS113: Inflammasome Primed Neutrophils Regulate Myocardial Infarction Induced Myelopoiesis
Authors: Sreejit Gopalkrishna , Nooti Sunil , Jaggers Robert , Johnson Jillian , Ma Jianjie , Murphy Andrew , Nagareddy Prabhakara R
Published: November 2022

Background:
Acute myocardial infarction (MI) results in overzealous production and infiltration of neutrophils to the ischemic heart. This is mediated in-part by granulopoiesis induced by the S100A8/A9-NLRP3-IL-1β signaling axis in injury-exposed neutrophils. Despite the transcriptional upregulation of the NLRP3 inflammasome and associated signaling components in neutrophils, the serum levels of IL-1β, the effector molecule in granulopoiesis was not impacted by MI suggesting that IL-1β is not released systemically. We hypothesize that IL-1β is released locally within the bone marrow (BM) by inflammasome-primed and reverse-migrating neutrophils.

Methods:
Using a combination of time-dependent parabiosis and flow cytometry techniques, we first characterized the migration patterns of different blood cell types across the parabiotic barrier. We next induced MI in parabiotic mice by permanent ligation of the LAD artery, and examined the ability of injury-exposed neutrophils to permeate the parabiotic barrier and induce granulopoiesis in non-infarcted parabionts. Finally, utilizing multiple neutrophil adoptive and BM transplant studies, we studied the molecular mechanisms that govern reverse migration and retention of the primed neutrophils, IL-1β secretion and granulopoiesis. Cardiac function was assessed by echocardiography.

Results:
MI promoted greater accumulation of the inflammasome-primed neutrophils in the BM. Introducing a time-dependent parabiotic barrier to the free movement of neutrophils inhibited their ability to stimulate granulopoiesis in the non-infarcted parabionts. Prior priming of the NLRP3 inflammasome is not a prerequisite, but the presence of a functional CXCR4 (C-X-C-motif chemokine receptor 4) on the primed neutrophils and elevated serum S100A8/A9 levels are necessary for homing and retention of the reverse-migrating neutrophils. In the BM, the primed neutrophils secrete IL-1β through formation of gasdermin D pores and, promote granulopoiesis. Pharmacological and/ or genetic strategies aimed at inhibition of neutrophil homing or release of IL-1β in the BM markedly suppressed MI-induced granulopoiesis and, improved cardiac function.

Conclusions:
Our data reveal a new paradigm of how circulatory cells establish a direct communication between organs by delivering signaling molecules (e.g., IL-1β) directly at the sites of action rather through systemic release. We suggest that this pathway may exist to limit the off-target effects of systemic IL-1β release
Abstract P1019:
CycleTrack
, A Genetic Method To Trace Cardiomyocyte Renewal In Small And Large Animal Models
Authors: Secco Ilaria , Backovic Ana , Vodret Simone , Dal Ferro Matteo , Tomczyk Mateusz , Gabisonia Khatia , Carlucci Lucia , Zentilin Lorena , Zacchigna Serena , Recchia Fabio A , Giacca Mauro
Published: November 2022

To what extent cardiac renewal occurs in adulthood and can be stimulated by therapeutic interventions is still a matter of investigation. In particular, over the last few years, significant effort has been made into awakening the endogenous regenerative potential of the adult mammalian heart after injury using different strategies. However, these studies are significantly hindered by the lack of tools to properly assess cardiomyocyte renewal over time. We have developed a genetic strategy, which we named CycleTrack, allowing a cumulative and accurate estimate of cardiomyocyte divisions in vivo. This method is based on the expression, in cardiomyocytes, of the Cre recombinase under the control of a 312-bp fragment of the Cyclin B2 promoter, which is specifically sensitive to cell proliferation. Replication-activated Cre acts on floxed GFP that is either present in the genome of transgenic mice or is exogenously delivered using Adeno-Associated Viral (AAV) vectors. As a result, cardiomyocytes traversing G2/M become irreversibly labeled. Using CycleTrack, we could monitor cardiomyocyte turnover in several physiological and pathological conditions. These included the measurement of the rate of proliferation of cardiomyocytes after apical resection in neonatal mice, the pro-regenerative effect of AAV9-mediated delivery of miRNA-199a and miRNA-590 after myocardial infarction in adult mice and the effect of pregnancy on myocardial hyperplasia. We also delivered the CycleTrack vectors into pig hearts to visualize cardiomyocyte turnover after myocardial infarction. Considering the large availability of Cre reporter mouse lines and the efficacy of AAVs to transfer genes into cardiomyocytes, we propose CycleTrack as a robust and straightforward tool to trace mitotic events for cardiac regeneration studies in small and large animal models.
Abstract P1043: Myocardial Infarction Induces Translocation Of Gut Bacteria To The Heart
Authors: Madaan Tushar , Nieman Michelle , Siddiqui Nabil , Lorenz John , Sadayappan Sakthivel , Kotagiri Nalinikanth
Published: November 2022

Introduction:
Myocardial hypoxia & intestinal hyperpermeability (IH) are two important events commonly observed in the aftermath of a myocardial infarction (MI) event. Recent studies have shown that gut bacteria and their metabolites are able to leak into the systemic circulation due to post-MI IH.

Hypothesis:
We hypothesized that facultative anaerobic gut bacteria such as
E. coli
will be able to translocate and colonize the hypoxic heart post-MI.

Methods:
C57 mice were given an oral administration of 10e9 cfu of luciferase/GFP expressing
E. coli
Nissle (EcN). Permanent occlusion of LAD arteries was performed to induce MI. Mouse hearts were checked for presence of EcN using PCR at different timepoints (n=32), IHC (n=4), and siderophore-based PET imaging (n=2). Immunoblotting of heart tissue lysate was used to check for presence of GFP.

Results:
PCR amplification of DNA extracted from MI hearts of mice given oral EcN revealed cardiac presence of translocated EcN. IHC analysis further confirmed presence of EcN in the heart. Siderophore-based PET imaging & radionuclide-based biodistribution studies revealed significantly higher uptake in the MI heart vs. Sham corroborating EcN presence. Immunoblotting revealed presence of GFP in cardiac tissue of MI mice suggesting that bacterial products and proteins could potentially translocate through the gut-heart axis.

Conclusion:
MI induced IH & myocardial hypoxia contribute to the translocation of gut bacteria and colonization of the ischemic heart.
Abstract P1050: Activation Of Bk
Ca
Channel Increases Myocardial Infarction After Ischemia-Reperfusion Injury In Neonatal Hearts
Authors: Sanghvi Shridhar , Szteyn Kalina , Ponnalagu Devasena , Sridharan Divya , Lam Alexander , Hansra Inderjot , Chaudhury Ankur , Majumdar Uddalak , Kohut Andrew , Rao Shubha , Khan Mahmood , Garg Vidu , Singh Harpreet
Published: November 2022

Large-conductance calcium and voltage-activated potassium channels (BK
Ca
) are heterogeneously expressed in a wide array of cells. In adult ventricular cardiomyocytes, activation of mitochondrial BK
Ca
channels is implicated in cardioprotection against ischemia-reperfusion (IR) injury. However, plasma membrane BK
Ca
channel activity has never been detected in adult ventricular cardiomyocytes. In this study, we report the presence of the BK
Ca
channel in the plasma membrane and mitochondria of neonatal murine and rodent cardiomyocytes. In neonatal cardiomyocyte (NCM), whole-cell potassium currents measured were sensitive to iberiotoxin (IbTx), suggesting the presence of BK
Ca
channels in the plasma membrane. Moreover, the open probability of single-channel recording decreased in presence of IbTx by 46+5% (n=10) and 32+5% (n=10) for rats and mice, respectively.
Ex vivo
studies with isolated neonatal hearts (p6) subjected to ischemia and post-conditioned with NS1619 during reoxygenation increased (58+3% as opposed to 48+5% for control, n>5 pups) the myocardial infarction whereas IbTx reduced (32+2%) the infarct size. In agreement, isolated NCM also presented increased apoptosis on treatment with NS1619 during hypoxia-reoxygenation (HR), whereas IbTx reduced TUNEL positive cells. In NCMs, activation of BK
Ca
channels increased the intracellular reactive oxygen species post HR injury. Electrophysiological characterization of NCMs indicated that NS1619 increased the beat period, field, and action potential duration, and decreased the conduction velocity and spike amplitude. In contrast, IbTx had no impact on the electrophysiological properties of NCMs. Taken together, our data established that inhibition of plasma membrane BK
Ca
channels in the neonates protects the heart/cardiomyocytes from IR injury. Furthermore, the functional disparity observed towards the cardioprotective activity of BK
Ca
channels in adults compared to neonatal heart could be attributed to their differential localization.
Abstract P1057: Implementing A Novel FRET-Based Myofilament Activation Biosensor To Interrogate Dynamic Cardiac Sarcomere Performance With Small Molecule And Genetic Manipulations During A Physiological Twitch
Authors: Martin Ashley A , Thompson Brian R , Metzger Joseph M
Published: November 2022

The sarcomere is the functional unit of cardiac muscle contraction. Understanding the precise mechanisms regulating this contraction is key to advancing insights into cardiomyopathies, which affect as many as 1 in 500 adults. Previous work has identified many key sarcomeric components and modulators necessary for the activation of contraction through inter-myofilament signaling. Although these studies have provided valuable information, the majority of this work has been done using
in vitro
protein recombination or permeabilized muscle. While informative, these systems lack significant physiological features of muscle, including load and intact excitation-contraction coupling mechanisms. Therefore, a large gap exists in the ability to monitor the time-sensitive stepwise physiological mechanism of muscle sarcomere activation in an intact system in real time. In order to further advance the study of sarcomere activation in both physiological and disease states, a cardiac specific myofilament-targeted FRET based biosensor has been designed and validated. The sarcomere activation biosensor reports myofilament dynamics during physiological twitch contractions in live cardiac muscle in real time in the form of simultaneous force and fluorescence measurements. This allows FRET-detection of the global conformational changes in TnC, a nexus point able to integrate many sarcomere-activating ligands, to be observed. From these changes in fluorescence the ensemble of events which lead to sarcomere activation and muscle contraction can be tracked. Data show regulatory ligands of myofilament activation in real time during a physiological twitch contraction. Currently, the biosensor is being utilized to investigate changes in sarcomere activation driven through the use of small molecules, including myosin inhibitor Mavacamten and TnC-TnI inhibitor W7, and those seen in MyBP-C knockout mice. The results of these investigations will be discussed.
Abstract P1063: Investigation Of Synthetic Membrane-stabilizing Copolymers To Dystrophin-deficient Skeletal Muscle Fibers
Authors: Hahn Dongwoo , Crabtree Adelyn , Lodge Timothy , Hackel Benjamin , Bates Frank , Metzger Joseph M
Published: November 2022

Duchenne muscular dystrophy (DMD) occurs by the lack of cytoskeletal protein dystrophin which is essential for the preservation of structural integrity of the muscle cell membrane. DMD develops severe skeletal muscle weakness, degeneration, and early death. We applied amphiphilic synthetic membrane stabilizers to the mdx skeletal muscle (flexor digitorum brevis; FDB) to compare the effectiveness in restoring contractile function in dystrophin-deficient muscle. After isolating the FDB fibers by enzymatic digestion and trituration from twenty-six adult male mice (5 C57/BL10, 21 mdx; at least 15 fibers/animal), we plated them on laminin-coated coverslip and treated them with poloxamer 188 (PEO
75
-PPO
30
-PEO
75
; 8,600 g/mol; P188), architecturally reversed (PPO
15
-PEO
200
-PPO
15
, 10,700 g/mol), and diblock (PEO
75
-PPO
15
-C
4
, 4,200 g/mol) copolymers in 15 and 150 μM at 25
o
C for 10 min. We assessed the twitch kinetics of sarcomere length (SL) and intracellular Ca
2+
transient by Fura-2, AM in field stimulation (25 V, 0.2 Hz, 25
o
C). Twitch peak SL shortening of mdx FDB fibers was depressed to 30% of the dystrophin-replete control FDB fibers from BL10 (P < 0.001). Treatment of mdx single FDB fibers with copolymers rapidly restored the twitch peak SL shortening by over 60%, especially P188 at 15 μM (60% of BL10 FDB, P < 0.001) and 150 μM (90% of BL10 FDB, P < 0.001). Twitch peak Ca
2+
transient from mdx FDB fibers was depressed by 50% compared to BL10 FDB fibers (P < 0.001). Interestingly, acute copolymer treatment of mdx FDB fibers significantly increased the twitch peak Ca
2+
transient by 20-45%, especially P188 at 150 μM up to 75% of the BL10 FDB fibers (P = 0.472). Our data shows that the structure and concentration of the synthetic copolymers lead to different effectiveness on rescuing the twitch contractile function and Ca
2+
handling in dystrophin-deficient skeletal muscle fibers. Notably, acute treatment for 10 min with copolymers (P188, 150 μM) elicited over 3-fold improvement of peak SL shortening in mdx isolated skeletal muscle fibers. This study highlights copolymers as effective membrane stabilizing molecules in the restoration of dystrophin-deficient skeletal muscle function and the importance of copolymer structure, molecular weight, and concentration.
Abstract P1076: Phosphodiesterase 1 (pde1) Modulates CAMP Signaling At The Sarcolemmal Membrane
Authors: Muller Grace K , Zaccolo Manuela , Kass David A
Published: November 2022

Phosphodiesterases (PDEs) regulate cyclic nucleotide-mediated cell signaling and offer a therapeutic means to leverage cardiac function. FDA-approved PDE3 inhibitors increase myocyte contractility by enhancing L-type calcium channel activity and sarcoplasmic reticulum (SR) calcium (Ca
2+
) load, but cause fatal arrhythmias in patients with heart failure. The PDE1 inhibitor ITI-214 is a novel inotrope that increases cardiac contractility in animal models and human patients with heart failure. Mechanistically, PDE1 inhibition leads to activation of the L-type Ca
2+
channel without altering the SR Ca
2+
load or myofilament-Ca
2+
sensitivity. The net result is enhanced myocyte contractility with comparatively less Ca
2+
increase and arrhythmias. PDE inhibition increases cAMP levels to activate protein kinase A (PKA), a pathway that regulates Ca
2+
handling and cycling. These results led us to hypothesize that PDEs 1 and 3 differ in localized domains where each controls cAMP-mediated signaling. Live-cell imaging experiments were performed in acutely isolated guinea pig myocytes expressing Förster resonance energy transfer (FRET) sensors to monitor changes in cAMP levels. These sensors were targeted to specific sarcolemmal domains or more generally to the cytosol. Our results showed that PDE1 inhibition is the predominant regulator of cAMP at the non T-tubular sarcolemmal membrane compared to other cAMP-hydrolyzing PDEs (PDE3 and 4). In contrast to PDE1, PDEs 3 and 4 were more indiscriminate and modulated cAMP in the cytosolic as well as in membrane domains. Upon beta-adrenergic stimulation, however, PDE4 but not PDEs 1 or 3 became the predominant cAMP modulator at the T-tubular membrane domain. These results provide evidence that PDE1 is a major regulator of sarcolemmal membrane cAMP, suggesting that PDE1 inhibitory effects upon the L-type Ca
2+
channel is localized to this domain. In future studies, we will examine if PDE1 inhibition selectively activates PKA in this region, and seek to identify the interacting proteins that support PDE1 modulation of sarcolemmal cAMP/PKA signaling. Resolving this cell signaling difference would provide mechanistic insight into a novel inotrope that has already completed a Phase II clinical evaluation in patients.
Abstract P1078: Early
Trypanosoma Cruzi
Infection Of Heart Endothelial Cells Modulates Hippo And Wnt Signaling Pathways In TSP-1 Dependent Manner
Authors: Rayford Kayla , Cooley Ayorinde , Lima Maria , Villalta Fernando , Pratap Siddharth , Nde Pius N
Published: November 2022

Trypanosoma cruzi
, an obligate intracellular protozoan parasite, is the etiological agent of Chagas Disease. Due to globalization, the disease is now present in all economically advanced countries of the world. About 40% of
T. cruzi
infected individuals will develop cardiac, neurological, and/or gastrointestinal pathology, with Chagasic Cardiomyopathy being the most prevalent pathology. During transportation, the molecular mechanisms by which the parasite signals and interact with host endothelial cells (EC), especially heart endothelium is currently unknown. Previous studies in our lab showed that the parasite dysregulates the expression of host genes, such as Thrombospondin-1 (TSP-1), to facilitate infection. Hippo and Wnt signaling pathways, in particular, have been implicated in cardiac development and disease pathogenesis. Here, we aim to evaluate the significance of TSP-1 in Hippo and Wnt signaling during the course of early
T. cruzi
infection. We challenged mouse heart endothelial cells (MHEC) from wild type (WT) and TSP1 knockout mice with
T. cruzi
and evaluated Wnt signaling by immunoblotting, YAP/β-catenin crosstalk via immunofluorescence assays, and parasite infectivity using β-catenin inhibitor. We found that in the absence of TSP1, the parasite induced the expression of Wnt-5a and enhanced the activation of GSK-3β simultaneously. In WT MHEC, the levels of Wnt-5a, p-GSK-3β were decreased compared to uninfected control. We also observed a continuous significant increase in the nuclear colocalization of β-catenin/YAP in TSP1 KO MHEC. Interestingly in WT MHEC, nuclear colocalization of β-catenin/YAP remained steady and reduced during later parasite challenge. Importantly, dysregulation of this crosstalk by pre-incubation of WT MHEC with a β-catenin inhibitor, endo-IWR 1, dramatically reduced the level of infection of WT MHEC. Parasite infectivity of inhibitor-treated WT MHEC was similar to the level of infection of TSP1 KO MHEC. These results indicate that TSP1 plays an important role in regulating Wnt/Hippo crosstalk and may be important potential therapeutic targets.
Abstract P1090: Polycystin-1 Regulates Calcium Handling And Contractility In Human IPSC-derived Ventricular Cardiomyocytes
Authors: Perez William , Hendrickson Troy , Altamirano Francisco J
Published: November 2022

Mutations in polycystin-1 (PC1) cause 85% of autosomal dominant polycystic kidney disease cases. This disease is characterized by progressive renal disease and multiple cardiovascular alterations. Evidence suggests that cardiovascular disease precedes the onset of renal failure. We recently demonstrated that PC1 ablation shortens cardiomyocyte action potential, decreases Ca
2+
transient amplitude, and reduces contractility in adult cardiomyocytes isolated from cardiomyocyte-specific PC1 knock-out mice compared to control mice. Alterations in Ca
2+
cycling in PC1-deficient cardiomyocytes were demonstrated to be due to action potential duration shortening and impaired sarcoendoplasmic reticulum ATPase activity. However, mouse and human cardiomyocytes have considerable differences in their action potential shapes and the ion channels involved. It remains unknown whether PC1 regulates these events in human cardiomyocytes. Here, we sought to investigate whether PC1 regulates excitation-contraction coupling in human ventricular cardiomyocytes. We differentiated commercially available human-induced pluripotent stem cell lines (hiPSC) into ventricular cardiomyocytes (hiPSC-CM) using published protocols (measurements were performed 60-90 days after differentiation). We developed a protocol to simultaneously measure action potentials (FluoVolt), intracellular Ca
2+
(Calbryte-590) and contractility using the IonOptix algorithm to detect changes in contrast as contractility measurements. PC1 ablation using siRNA decreased action potential duration and impaired Ca
2+
transients and contractility in hiPSC-CMs. Because hiPSC-CMs have several limitations due to their immature state, we employed a well-validated method to improve their maturation that utilizes a Matrigel mattress and hormone supplementation (dexamethasone; Dexa / thyroid hormone; T3; PMID: 28974554; developed by Dr. Bjorn Knollmann’s lab). Using the Matrigel mattress/Dexa/T3 technique, we increased hiPSC-CM maturation measured as follows: cardiomyocytes were oriented and contracted in the longitudinal axis, ryanodine receptor type 2 and alpha-actinin localized with a striated pattern, exhibited robust inward K
+
rectifier currents (I
K1
) and fired action potentials that resemble human adult cardiomyocytes. In this model, PC1 knockdown also decreased action potential duration and impaired Ca
2+
transients and decreased contractility measured as fractional shortening. Altogether, this evidence shows for the first time that PC1 regulates human cardiomyocyte physiology and suggests that cardiac alterations in patients with autosomal dominant polycystic kidney disease may emerge as a result of impaired cardiomyocyte contractile function.