bluuebunny/crossref_metadata_2025 huggingface dataset:
Does Action Observation of the Whole Task Influence Mirror Neuron System and Upper Limb Muscle Activity Better Than Part Task in People With Stroke?
Authors: Sulfikar Ali A. , Bhat Mayur , Palaniswamy Hari Prakash , Ramachandran Selvam , Kumaran Senthil D.
Published: October 2024

Background: Task‐based action observation and imitation (AOI) is a promising intervention to enhance upper limb (UL) motor function poststroke. However, whether whole/part task must be trained in the AOI therapy needs further substantiation.

Objective: The objective of this study is to assess and compare the mirror neuron activity and UL muscle activity during AOI of reaching task in terms of whole task (complete movement) and part task (proximal arm movements and distal arm movements).

Methods: In this cross‐sectional study, 26 participants with first‐time unilateral stroke were asked to observe the prerecorded videos of a reaching task in terms of a whole task and proximal and distal components, followed by imitation of the task, respectively. Electroencephalographic (EEG) mu rhythm suppression and electromyographic amplitude of six UL muscles were measured during the task.

Results: The analysis of EEG revealed a statistically significant mu suppression score, indicating mirror neuron system activity, during AOI of the whole task in C3 (p = <0.001) and C4 (p = <0.001) electrodes compared to the part task. Percentage maximum voluntary contraction amplitudes of the deltoid (p = 0.002), supraspinatus (p = <0.001), triceps brachii (p = 0.002), brachioradialis (p = 0.006), and extensor carpi radialis (p = <0.001) muscles showed a significant increase in muscle activity during AOI of the whole task. Also, there seems to be a task observation–specific activation of muscles following AOI of proximal or distal tasks.

Conclusion: The practice of the whole task should be given emphasis while framing the AOI treatment module to enhance reaching in people with stroke.

Trial registration: Clinical Trials Registry‐India (CTRI) identifier: CTRI/2018/04/013466.
Physical Activity, Cardiorespiratory Fitness, and Atherosclerotic Cardiovascular Disease: Part 2
Authors: Franklin Barry A. , Jae Sae Young
Published: October 2024

Background: In this second section of our 2-part review on the role of physical activity (PA) and cardiorespiratory fitness (CRF) in preventing and treating atherosclerotic cardiovascular disease (CVD), we expand on topics covered in part 1, including a comparison of moderate-intensity continuous training versus high-intensity interval training, the beneficial role of PA and CRF in heart failure, potential mal-adaptations that may result from extreme endurance exercise regimens, and the incidence of cardiac arrest and sudden cardiac death during marathon running and triathlon participation. Further, we review the principles of exercise prescription for patients with known or suspected CVD, with specific reference to exercise modalities, contemporary guidelines, the minimum exercise training intensity to promote survival benefits, and long-term goal training intensities, based on age-, sex-, and fitness-adjusted targets. Finally, we provide practical “prescription pearls” for the clinician, including a simple rule to estimate metabolic equivalents (METs) during level and graded treadmill walking, research-based exercise training recommendations, using steps per day, MET-minutes per week, and personal activity intelligence to achieve beneficial treatment outcomes, as well as the heart rate index equation to estimate energy expenditure, expressed as METs, during recreational and leisure-time PA. Summary: This review compares moderate-intensity continuous training and high-intensity interval training, examines the role of PA and CRF in managing heart failure, and discusses the cardiovascular risks associated with extreme endurance exercise. It also provides practical guidelines for exercise prescription tailored to patients with CVD, highlighting advanced exercise prescription strategies to optimize cardiovascular health. Key Messages: Physicians and healthcare providers should prioritize referring patients to home-based or medically supervised exercise programs to leverage the cardioprotective benefits of regular PA. For most inactive patients, an exercise prescription is essential for improving overall health.
Incidental Uptake in Gastrointestinal Tract on 18F-FDG-PET/CT: Is It Worth to Investigate? A Study with 371 Patients
Authors: Correia Gomes Luís , Fraga Davide , Lage Pedro , Salgado Lucília , Claro Isabel
Published: October 2024

Background and Study Aims: Positron emission tomography/computed tomography (PET/CT) with 2-[18F]FDG (FDG) has been increasingly used to detect or monitor neoplasms. Gastrointestinal tract (GIT) is one of the most common sites of FDG uptake, leading to increasing requests for endoscopic examinations. We aimed to evaluate the nature and significance of unexpected PET/CT-FDG findings in the GIT. Patients and Methods: We retrospectively analyzed 371 consecutive patients with incidental GIT findings on PET/CT-FDG between June 2016 and October 2021 who were subsequently referred to endoscopic examinations. Demographic data, PET/CT-FDG results, endoscopic findings, and histological analysis were analyzed. Results: Of 194 colonic incidental uptakes, 102 (52.6%) corresponded to at least premalignant lesions, being 57 (29.4%) advanced adenomas and 23 (11.9%) adenocarcinomas. Of 193 upper GIT incidental uptakes, there were 11 (13.8%) esophageal and 14 (14.4%) gastric malignant/premalignant lesions. The maximum standardized uptake value (SUVmax) significantly varied according to the nature of the lesion, being higher in malignant lesions (in the esophagus, stomach, and colon). However, an optimal SUVmax cutoff was only found for stomach (SUVmax 8.2; sensitivity of 79% and specificity of 76%). There was a significant association between the site of uptake and the nature of the lesion – left colon and gastric body uptake were associated with neoplastic origin whereas rectum and lower esophagus were associated with inflammatory or no endoscopic changes. Conclusions: Any incidental uptake in the lower GIT should be investigated provided that patients are suitable for further treatment. However, in the upper GIT the characteristics of uptake on 18F‐FDG‐PET/CT may allow to select those who need endoscopic examination.
Runs of Homozygosity Decipher Genetic Diversity in Cattle Breed Dwelling in the Colder Regions of the World
Authors: Mahar Karan , Goli Rangasai Chandra , Chishi Kiyevi G. , Ganguly Indrajit , Dixit S.P. , Singh Sanjeev , Choudhary Sonu , Rathi Pallavi , Chinnareddyvari Chandana Sree , Diwakar Vikas , Metta Muralidhar , Prabhu Immanual Gilwax , Kumar Amit , Sarkar Soumajit , Sukhija Nidhi , Kareningappa Kanaka Krishnamurthy
Published: October 2024

Background: Our study focuses on Yakutian cattle, a Siberian native breed, examining its inbreeding and diversity through genome-wide analysis of runs of homozygosity (ROHs). Yakutian cattle are adapted to Siberia’s harsh sub-arctic conditions, enduring temperatures below −70°C. However, the population genetics studies on this breed are scanty, to document the genetic uniqueness in these cattle. Results: We analyzed 40 Yakutian cattle with strict quality control for ROH detection yielding 683 homozygous segments, averaging 17 per individual with an average length of 9 Mb. ROH regions were found to be involved in important pathways pertaining to cold adaptation. Autozygosity ranged from 1% to 12% of the genome, with a relatively low average inbreeding coefficient (FROH) of 0.057, as compared to other breeds. Also, the different diversity indicators, namely, principal component analysis, heterozygosity, and effective population size analysis, revealed the prevalence of genetic diversity within the breed. Conclusion: Our findings on ROH are the first of its kind in Yakutian cattle that support their adaptability to colder environments, as evidenced by low inbreeding and high genetic diversity.
Prati ciências: material digital de apoio a professores de ciências
Authors:   Universidade do Porto Centro de Investigação em Biodiversidade e Recursos Genéticos, Ramos Ana Luisa , Bonzanini Taitiany Karita ,   Universidade de São Paulo. Escola Superior de Agricultura Luiz de Queiroz
Published: October 2024

O material apresenta roteiros de atividades práticas para aulas de Ciências no Ensino Fundamental II, que possam ser realizadas com materiais de fácil acesso e procedimentos simples, não sendo necessários laboratórios para sua realização. Resultou de uma pesquisa de conclusão de curso, desenvolvida na Licenciatura em Ciências Biológicas da ESALQ.
Abstract Mo020: Loss of TRAF2 Signaling Mediates Mitochondrial Dysfunction in Doxorubicin-Cardiomyopathy
Authors: Kirshenbaum Lorrie , Dhingra Rimpy , Rabinovich-Nikitin Inna , Gang Hongying , Margulets Victoria , Diwan Abhinav , Javaheri Ali
Published: October 2024

Rationale:
Cytokines such as TNFα have been implicated in cardiac dysfunction and toxicity associated with doxorubicin (DOX). While TNFα can elicit different cellular responses including survival or death, the mechanisms underlying these divergent outcomes in the heart remains cryptic. The E3 ubiquitin ligase TRAF2 provides a critical signaling platform for K63 - linked ubiquitination of RIPK1, crucial for NF-kB activation by TNFα. Whether alterations in TNFα-TRAF2 signaling underlie the cardiotoxic effects of DOX, remains poorly understood.

Results:
In contrast to vehicle treated mice, severe ultrastructural defects including cytoplasmic swelling, mitochondrial perturbations, and elevated TNFa levels were observed in the hearts of mice treated with DOX. While investigating the involvement of TNFa in DOX cardiotoxicity, we discovered that in the absence of DOX, NF-kB was readily activated by TNFa. However, TNFα -mediated NF-kB activation was impaired in cardiac myocytes treated with DOX. This coincided with loss of K63- linked poly-ubiquitination of RIPK1, attributed to the proteasomal degradation of TRAF2. Further, TRAF2 protein expression was markedly reduced in hearts of cancer patients treated with DOX. Impaired TRAF2 signaling resulted in the activation of Bnip3 and mitochondrial perturbations, including disrupted bioenergetics, loss of membrane potential and permeability transition pore opening. We further established that the reciprocal actions of the ubiquitinating and de-ubiquitinating enzymes c-IAP1 and USP19 respectively regulated the proteasomal degradation of TRAF2 in DOX treated cardiac myocytes. Importantly, an E3 ligase mutant of c-IAP1(c-IAP1 H588A) or gain of function of USP19, prevented proteasomal degradation of TRAF2 and DOX -induced cell death. Further, wild type TRAF2 but not a RING finger mutant defective for K63 ubiquitination of RIPK1, restored NF-kB signaling and suppressed DOX-induced cardiac cell death. Finally, cardiomyocyte-restricted expression of TRAF2 (AAV9-TRAF2)
in vivo
protected against mitochondrial defects and cardiac dysfunction induced by DOX.

Conclusions:
Our findings reveal a novel signaling axis that functionally connects the cardiotoxic effects of DOX to proteasomal degradation of TRAF2. Disruption of the critical TRAF2 survival pathway by DOX, sensitized cardiac myocytes to TNFa mediated necrotic cell death.
Abstract Mo081: Genetic Deletion of Histone Lysine Demethylase KDM5A in Cardiomyocytes Attenuates LMNA-associated Dilated Cardiomyopathy
Authors: Deogharia Manisha , Marian Ali , Gurha Priyatansh
Published: October 2024

Background:
Heart failure (HF) is a major cause of morbidity and mortality. Hereditary cardiomyopathies are the prototypic forms of HF. Notable among them is dilated cardiomyopathy (DCM) caused by mutations in the
LMNA
gene encoding Lamin A protein (LMNA-DCM). We have recently shown that KDM5A, a histone H3 lysine 4 demethylase, is activated in the hearts of human patients and mouse models of LMNA-DCM. Inhibition of the KDM5 family of proteins activates gene expression, including expression of genes involved in oxidative phosphorylation (OXPHOS) and fatty acid oxidation (FAO) in iPSC-derived cardiomyocytes. Thus, we hypothesized that activation of the KDM5A in the LMNA-DCM is pathogenic and contributes to cardiac dysfunction.

Objective:
The objective of this study is to determine the role of KDM5A in the pathogenesis of LMNA-DCM.

Methods and Results:
The
Lmna
and
Kdm5a
genes were concomitantly deleted in the cardiac myocytes (CMs) in mice using cre recombinase. CM-specific deletion of the
Lmna
gene (
Myh6-Cre:Lmna
F/F
), representing a mouse model of LMNA-DCM, led to severe cardiac dysfunction, increased apoptosis, and premature death as published. The
Myh6-Cre:Kdm5a
F/F
mice showed no discernible phenotype. Deletion of the
Kdm5a
gene in CMs in the LMNA-DCM mice
(Myh6-Cre:Lmna
F/F
:Kdm5a
F/F
) attenuated myocardial apoptosis, improved cardiac function, and prolonged survival. Specifically, left ventricular end-systolic and end-diastolic diameters (LVESD and LVEDD, respectively) were smaller and LV fractional shortening (LVFS) was greater in the
Myh6-Cre: Lmna
F/F
:Kdm5a
F/F
mice as compared to the

Myh6-Cre:Lmna
F/F

mice. Likewise, the median and maximum survival times were increased by ~ 40 to 50% in the
Myh6-Cre:Lmna
F/F
:Kdm5a
F/F
. Analysis of the CM transcripts by RNA-sequencing showed dysregulation of 6609 genes in the
Myh6-Cre: Lmna
F/F
CMs and 2791 genes were attenuated in the
Myh6-Cre:Lmna
F/F
:Kdm5a
F/F
. Pathway analysis showed that genes involved in FAO and OXPHOS were the most suppressed in LMNA-DCM and were rescued upon
Kdm5a
deletion.

Conclusions:
Deletion of the
Kdm5a
gene in the LMNA-DCM mice imparts salubrious effects by prolonging survival and improving cardiac dysfunction, and upregulating genes involved in OXPHOS and FAO. Thus, activation of KDM5A is likely pathogenic and contributes to the phenotypic expression of LMNA-DCM.
Abstract Tu002: Mesenchymal Stem Cell-Derived Extracellular Vesicles Mitigate Mitochondrial DNA-Induced Activation of Porcine Peripheral Blood Mononuclear Cells
Authors: Zahra Sumbule , Rolland Tyler , Cucinotta Daniel , Weil Brian
Published: October 2024

Objective:
Systemic inflammation is a well-established component of post-cardiac arrest syndrome (PCAS), a condition responsible for significant morbidity and mortality in patients that are initially resuscitated from sudden cardiac arrest. Immune cell activation is pivotal in PCAS pathophysiology, with emerging evidence implicating mitochondrial DNA (mtDNA) as a potent pro-inflammatory stimulus in this context. Given the paucity of available interventions to inhibit mtDNA-induced immune cell activation, we investigated the therapeutic potential of mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) to modulate the inflammatory response of porcine peripheral blood mononuclear cells (PBMCs) activated by mtDNA
in vitro
.

Methods:
Porcine bone marrow-derived mesenchymal stem cells (MSCs; P2) were cultured for 5 days and serum-starved for 3 days to collect MSC-EVs. These EVs were isolated from conditioned media (CM) using size exclusion chromatography and ultracentrifugation, then characterized by nanoparticle tracking analysis (ZetaView) and ExoCheck. Naïve PBMCs from healthy swine were cultured for 1 week before stimulation with lipofectamine 2000 (TR-control) or 2 µg/mL mtDNA+TR. MSC-EVs were added to activated PBMCs (2.5e8–2e9 EVs/well) and inflammatory surface marker expression (flow cytometry), inflammatory cytokine transcripts (qPCR) and release (ELISA), and reactive oxygen/nitrogen species (ROS/RNS) production were assessed 24 hours later.

Results:
mtDNA-stimulated PBMC populations exhibited altered surface marker expression consistent with enrichment of inflammatory granulocytes (CD172+CD163-), macrophages (CD14+CD163+), and adhesion molecules (CD172+) compared to TR-control (all p<0.05). Addition of MSC-EVs significantly reduced the number of all inflammatory cell populations, with the most prominent effects achieved at the highest EV dose (2e9 EVs/well). Likewise, MSC-EVs significantly attenuated mtDNA-activated PBMC expression and secretion of TNFα, IL-1β, and IL-6 (
Figure)
and decreased ROS/RNS production in a dose-dependent manner.

Conclusion:
MSC-EVs attenuate mtDNA-induced activation of PBMCs in a dose-dependent manner, as determined by reductions in pro-inflammatory surface marker expression, inflammatory cytokine gene expression and secretion, and ROS/RNS production. These findings indicate that MSC-EVs may be a viable therapeutic for the inhibition of mtDNA-mediated immune activation in PCAS.
Abstract Tu036: Adipose tissue plasticity in response to early pathological stress on the heart and mediation by adipose thermogenic activation
Authors: James Natasha Maria , Challa Azariyas , Vidal Souza Pablo , Pardeshi Parth , Maurya Santosh , Wang Yang , Baer Lisa , Carley Andrew , Lewandowski E Douglas , Stanford Kristin
Published: October 2024

Background:
Cardiovascular diseases (CVD) are the leading cause of death globally, accounting for approximately 17.9 million mortalities per year. Previous studies have shown that there is an early response of white and brown adipose tissue (WAT and BAT, respectively) to pathological stress on the heart prior to cardiac dysfunction. Interestingly, human subjects with detectable BAT activity (a thermogenic tissue) exhibit a reduced risk of Congestive Heart Failure, but the mechanism for this is unclear. We hypothesized that increasing the thermogenic acitivity of adipose tissue would be beneficial during the initial cardiac hypertrophic signaling prior to the development of compensatory hypertrophy.

Methods:
We studied both WAT and BAT in male and female C57BL/6J mice subjected to cardiac pressure overload via transaortic constriction (TAC). 24 hours post-surgery, mice were subjected to room temperature (RT) 22° C or mild cold 16°C to stimulate BAT activation for 48 hours after which tissues were collected. The study groups included: SHAM RT, TAC RT, SHAM COLD, TAC COLD. We used uncoupling protein 1 knockout (UCP1-KO) mice, with impaired thermogenic activity as a negative control.

Results:
At 72 hours post tac surgery, expression of thermogenic and metabolic genes remained unchanged in BAT and WAT of both sexes. However, mild cold exposure resulted in an increase in
UCP1
expression in BAT of male, but not female mice while other metabolic and thermogenic gene markers’ expression was increased in a sex-specific manner. Adaptations to scWAT were more moderate than BAT, with an increase in
Ucp1
after cold exposure in both sexes. Moreover, 48 hours of cold exposure attenuated expression of gene markers for hypertrophic signaling in the heart during TAC. Importantly, these adaptations were absent in UCP1-KO mice, signifying that intact thermogenic activity in adipose tissue was required for the attenuated expression of hypertrophic markers in the heart. Interestingly, cardiac fibrosis markers were downregulated with TAC-Cold in male mice only, suggesting a stronger response compared to female mice.

Conclusions:
Our findings demonstrate that pathological stress on the heart during TAC does not impact adipose tisse thermogenesis. Additionally, activating thermogenic gene expression of adipose tissue via cold exposure mediates the early induction of hypertrophic signaling in the heart within 72 hours of TAC. Notably, adipose tissue plasticity is sex-specific.
Abstract Tu058: Reverse Remodeling with LVAD Therapy is Associated with Alternative Splicing of CAMK2D
Authors: Martin Thomas , Hunt Dakota , Ebmeier Christopher , Ambardekar Amrut , buttrick peter , leinwand leslie
Published: October 2024

Therapies for end-stage heart failure are lacking and the number of patients awaiting heart transplants far outweighs availability. Left ventricular assist devices (LVADs) take over cardiac pump function and are used as a bridge-to-transplant. Notably, some patients on LVADs experience recovery of cardiac function and reversal of pathological cardiac remodeling; however, the molecular mechanisms underlying these favorable outcomes are unknown. In this study, we studied 19 paired pre- and post-LVAD samples with 10 clinically categorized as responders and 9 as non-responders. Bulk mRNA sequencing identified 486 differentially expressed genes post-LVAD, including reduced pro-inflammatory gene expression and increased expression of genes involved in metal ion binding; however, just 23 total genes were differentially expressed between responders and non-responders. To determine whether LVAD differentially impacted the proteome in responders and non-responders, we next performed quantitative proteomics on post-LVAD samples. This analysis identified 154 differentially expressed proteins and revealed that factors involved in mRNA processing were higher expressed in LVAD responders. Re-analysis of our mRNA sequencing data for gene isoforms identified that alternative splicing of calcium and calmodulin-dependent protein kinase 2-delta (
CAMK2D
) was the most significantly impacted event with non-responders having increased inclusion of exon 14 (denoting the B isoform), which encodes a nuclear localization sequence. Notably, there was a strong negative correlation between exon 14 inclusion and ΔLVEF post-LVAD. We validated these findings using targeted exon-specific quantitative PCR, which confirmed that expression of

CAMK2D
B

was higher both pre- and post-LVAD in non-responders, while responders had post-LVAD levels matching non-failing controls. We next investigated whether responders and non-responders differed at the phospho-proteome level via mass spectrometry and found three serines in CaMK2δ exon 14 were the only differentially abundant phosphosites and all increased in non-responders. Localization studies
in vitro
with adenoviruses expressing phospho-null or phospho-mimetic CaMK2D
B
constructs revealed that these phosphorylation events precluded nuclear localization. In conclusion, we identified differential alternative splicing and phosphorylation of CaMK2δ in LVAD responders, which may represent novel therapeutic targets for heart failure recovery.
Abstract Tu079: The Calcium Handling Machinery and Electrophysiology is Remodeled in Friedreich’s Ataxia
Authors: Czornobil Roman , Abu-Assail Obada , Wood Elizabeth , Lynch David , Noujaim Sami , Chidipi Bojjibabu
Published: October 2024

Background:
Friedreich's ataxia (FA) is a progressive neurodegenerative disorder. FA is caused by a deficiency in the frataxin protein due to an expansion of GAA repeats in the first intron of the frataxin gene. Reduced frataxin protein expression is thought to negatively affect mitochondrial function, leading to increased oxidative damage. Although FA is considered a neurodegenerative disorder, it is not completely understood why most patients present with left ventricular systolic dysfunction and die from cardiac events.

Objective:
The objective of our study is to determine whether abnormal calcium handling is a molecular mechanism of cardiac dysfunction in FA.

Methods:
We used the frataxin knock-out (KO) mouse model of FA as well as human heart samples from donors with FA and from unaffected donors. The expression of calcium-handling proteins was assessed using proteomics and westernblot. We used echocardiography and telemetric ECG to assess cardiac function in the mice. We also performed in-vivo programmed electrical stimulation (PES) to assess ventricular arrhythmogenesis in the mouse heart.

Results:
Proteomics and western blot showed that SERCA2, Ryr2, PLN, CASQ2, and CaMKII were downregulated and that the phosphorylation of Ryr2 and PLN (Ser16 and Thr17) were also significantly decreased in left ventricular tissue from humans with FA and from KO mice. The mitochondrial calcium handling proteins VDAC and MCU were upregulated in left ventricular tissue from humans with FA and from KO mice. On telemetric ECG, the RR tended to prolong in the KO animals compared to wild types (p=0.053), and heart rate variability analysis indicated heightened cardiac parasympathetic input. Echocardiography showed that the ejection fraction and fractional shortening were significantly decreased and left ventricular wall thickness and diameter were significantly increased in the KO mice. In PES, ventricular arrhythmogenesis was greater in KO mice compared to wild types.

Conclusions:
The reduced expression of calcium handling proteins may contribute to cardiac contractile and electrophysiological abnormalities in FA.

Keywords:
Friedreich's ataxia, SERCA2, Ryr2, PLN, CASQ2, and CaMKII.
Abstract Tu107: Decreased Mitochondrial Protein Expression Accompanies Atrial Hypertrophy but Precedes Atrial Fibrillation Onset in a Mouse Model of Spontaneous Atrial Fibrillation
Authors: Li Ling , Hanmer Shannon , Willard Belinda , Smith Jonathan , Laurita Kenneth , Chung Mina , Schumacher Sarah , Barnard John , Van Wagoner David , Rennison Julie
Published: October 2024

Introduction:
Atrial fibrillation (AF), characterized by episodes of rapid and irregular heart rate, increases left atrial (LA) demand for ATP. While the heart is metabolically flexible, able to use fatty acids, lactate, glucose, ketones, and amino acids to generate ATP, fatty acids are the primary fuel source. AF is associated with altered mitochondrial structure and function, but the role of energetic pathways in early stages of AF progression is not clear.

Hypothesis:
Decreased expression of mitochondrial proteins contributes to AF progression.

Aim:
Evaluate expression of proteins that influence mitochondrial structure and energetic capacity in AF onset and progression.

Methods:
AF burden was assessed weekly by electrocardiography (ECG) in male wild-type (WT) and cAMP responsive element modulator IbΔC-X heterozygous mice (CREM) (n=3). LA (4 - 9 mg) was obtained at 7 weeks and 16 weeks. Global proteomics was conducted, and 139,316 peptides were identified that mapped to 6000 robustly quantified proteins.

Results:
LA hypertrophy was evident in 7-week and 16-week CREM mice (Figure 1). We detected 2408 proteins differentially expressed in CREM compared to WT. At 7 weeks, Hallmark pathways adipogenesis, fatty acid metabolism, oxidative phosphorylation, and myogenesis were decreased in CREM (Table 1). These changes persisted at 16 weeks. We identified 740 differentially expressed mitochondrial proteins using MitoCarta 3.0 (552 decreased, 188 increased). Mitochondrial pathways associated with energy production, including branched chain amino acid metabolism, fatty acid transport and oxidation, carbohydrate metabolism, and tricarboxylic acid cycle were decreased (Table 1). Mitochondrial cristae organizing system (MICOS) and mitophagy, pathways that influence mitochondrial structure and content, were decreased. Terminal ECGs confirmed AF in 16-week CREM mice only. The coefficient of variation of RR intervals (CVRR), a measure of heart rate variability, was increased in 16-week CREM (Figure 2). These data suggest that decreased expression of mitochondrial proteins accompanies LA hypertrophy but precedes onset of AF.

Conclusions:
Decreased expression of proteins that regulate mitochondrial structure and energy production prior to AF onset suggests that these proteins play an important role in early LA changes that promote AF onset and progression. Future studies will examine how these changes in protein expression relate to alterations in atrial energy homeostasis.
Abstract Tu139: Ferroptotic Cardiomyocytes Support Angiogenesis in the Infarcted Myocardium
Authors: Stairley Rebecca , Trouten Allison , Li Shuang , Roddy Patrick , Deleon-Pennell Kristine , Sucov Henry , Liu Chun , Tao Ge
Published: October 2024

Introduction:
Mammalian cardiomyocytes have a low turnover rate which is insufficient to repopulate the damaged myocardium after injury caused by myocardial infarction (MI). Previously, we reported that ferroptosis is the main mechanism of cardiomyocyte death in the infarcted neonatal heart. We hypothesized that ferroptotic cardiomyocytes play a beneficial role during the wound healing process.

Aims:
In this study, we aim to characterize the role of ferroptotic cardiomyocytes by investigating their influence on non-cardiomyocyte populations in the heart post-MI.

Methods:
Left anterior descending artery occlusion (LAD-O) was performed on wild type (C
5
7BL/6J x FVB) mice at P1. Ferroptosis inhibitor ferrostatin-1 (Fer-1) was delivered at a 2mg/kg dose via subcutaneous injection immediately following LAD-O procedure and again daily until P4. Histology and immunofluorescence were performed to assess myocardial regeneration. iPSC-derived cardiomyocytes (iCMs) were treated with Erastin, Staurosporine, or vehicle control to generate conditioned media. HUVECs were treated with iCM conditioned media for tube formation assays.

Results:
The infarcts of Fer-1 treated animals show decreased total macrophages from 5.5% to 0.16% (P < 0.05, N = 6 per group, S.E.M. = 1.2%) and those present were less polarized towards the M2 phenotype, decreased from 62.5% to 7.9% (N = 6 per group, P < 0.001, S.E.M. = 9.6%). This trend was confirmed via flow cytometry. The number of Emcn
+
cells in the infarct was significantly reduced in Fer-1 treated mice, decreasing from 17% to 10% (S.E.M. = 1.7%, N = 4 per group, P < 0.01). HUVECs treated with conditioned media generated by treating iPSC-derived cardiomyocytes with Staurosporine, but not Erastin or DMSO, showed reduced mesh area (fold change from 0.94 to.0 24, N = 5 per group, P < 0.01. S.EM. = .128). Other tube formation endpoints such as extremities, junctions, and branching followed this trend.

Conclusions:
Our analysis revealed that ferroptotic cardiomyocytes play a supportive role in the wound healing and tissue remodeling environment by directly supporting angiogenesis and polarizing the cardiac macrophages towards a more pro-angiogenic state.
Abstract Tu143: Hsp90β Facilitates Cell Death During Acute Cardiac Ischemia/Reperfusion Injury
Authors: Roberts Richard , Wisner Halley , Sen Sujoita , Lam Chi
Published: October 2024

Myocardial infarction (MI), which caused 110,000 deaths in America in 2020, occurs following a complete blockage (ischemia) in one or more of the coronary arteries. Additionally, clearing the blockage (reperfusion) causes massive cell death, resulting in permanent tissue damage in the heart. Preventing cell death in IR injury is key to improving patient outcome in this disease. Molecular chaperones have emerged as promising targets in IR injury due to their pro-survival functions. However, the role of Hsp90β, the most abundant and constitutively active chaperone, in IR injury has not been fully elucidated. Therefore, we generated a cardiac-specific Hsp90β conditional knockout (cKO) mouse model to address it. To our surprise, current preliminary results suggest that cardiac-specific Hsp90β ablation reduces myocardial damage during IR injury, as Hsp90β cKO mice showed reduced infarct size compared to control. Opening of the mitochondrial permeability transition pore (mPTP) is the major driver of necrotic cell death during early reperfusion. Isolated mitochondria from the hearts of Hsp90β cKO mice show decreased sensitivity to mPTP opening during calcium overload, suggesting that it may be one of the cardioprotective mechanisms mediated by Hsp90β ablation. The Hsp90β cKO hearts also display an anti-apoptotic shift of the BAX/BCL2 apoptotic rheostat, suggesting the heart will be more resistant to apoptotic cell death during late reperfusion phase as well. Findings from this study will help us understand the role of Hsp90β in cell death activation during IR injury. In the future, we plan to investigate the underlying protective mechanisms mediated by Hsp90β ablation.
Abstract We064: Investigating the Cardioprotective Role of Neutrophil-Specific STING in Myocardial Ischemia/Reperfusion Injury
Authors: Brockman Maegan , Wang Lanfang , Scruggs Triniti , Calvert John , Levit Rebecca
Published: October 2024

Introduction:
Inflammation following myocardial ischemia/reperfusion injury (MI/R) plays a significant role in damaging cardiomyocytes and influencing infarct size, with neutrophils being the most rapid and numerous cells recruited to the myocardium post reperfusion. Single-cell RNA sequencing data identified neutrophils with upregulated type I interferon (IFN) signaling in the hearts of mice 24 hours post-MI/R.

Aim:
This study aims to address how the type I IFN neutrophil phenotype identified affects inflammation in MI/R. We hypothesize that neutrophils with upregulated type I IFN signaling are cardioprotective in MI/R.

Methods:
Male neutrophil-specific STING knockout (KO) mice underwent 60 minutes of ischemia followed by reperfusion for 24 hours. Immune cells were isolated from the left ventricle for flow cytometry. Subsequently, mice underwent 60 minutes of ischemia followed by reperfusion, and cardiac function was assessed two weeks-post reperfusion.

Results:
Flow cytometry revealed a significant decrease in the number of neutrophils in the myocardium of neutrophil-specific STING KO mice compared to wildtype mice after 24 hours of reperfusion (5.7x10
4
± 1.0x10
4
; 8.7x10
4
± 1.9x10
4
, Two-way ANOVA, N=5-6, p=0.0006). Neutrophil-specific STING KO mice showed significant reduction in ejection fraction (29.2% ± 6.4) compared to wildtype mice (38.3% ± 6.7; Two-way ANOVA, N=4-5, p=0.036), and significantly increased left ventricular end systolic volume (4.39mm ± 0.48) compared to wildtype mice (3.59 ± 0.62; Two-way ANOVA, N=4-5, p=0.017), indicating impaired cardiac function in knockout mice.

Conclusion:
These data suggest that neutrophil STING plays a cardioprotective role in MI/R. These studies provide the foundation to further investigate the cardioprotective mechanisms behind neutrophil-specific STING. Identifying a novel cardioprotective mechanism could be a clinically feasible point of intervention to improve recovery after MI/R.
Abstract We075: Stimulator of Interferon Genes (STING) Regulates T Cell Fate in Cardiometabolic Heart Failure with Preserved Ejection Fraction (HFpEF)
Authors: Smolgovsky Sasha , Bayer Abraham , Emig Ramona , Magri Zoie , aronovitz mark , kaur Kuljeet , Poltorak Alexander , Blanton Robert , Alcaide Pilar
Published: October 2024

Introduction:
Combined obesity and hypertension alter the endoplasmic reticulum (ER) stress response by downregulating X-box binding protein 1 (
Xbp1
) in T cells, which heightens T cell inflammatory potential and persistence in Heart Failure with Preserved Ejection Fraction (HFpEF), a prevalent syndrome with no cure. The ER protein Stimulator of Interferon Genes (STING), in concert with
Xbp1
, coordinates T cell inflammation and death in other contexts, yet its role in the T cell inflammatory response in cardiometabolic HFpEF is unknown.

Hypothesis:
STING activation and dysregulated ER stress responses drive T cell inflammation in HFpEF by balancing T cell activation and persistence in response to combined comorbidities.

Methods:
C67BL/6J (wild type, WT) mice were fed high-fat diet/L-NAME (H/L) or standard chow (STD) for 5 weeks, and STING activation was measured in splenic CD4
+
T cells by Western blotting. Splenic CD4
+
T cells from WT or STING-deficient mice (STING
-/-
) were activated with αCD3/αCD28 for 3 days
in vitro
, then treated with complete media (CM) or CM with 0.2 mM palmitate (abundant in dietary fat)/0.2 mM L-NAME (PALNA) for up to 48 hours. T cell gene expression, activation, and death were assessed using qPCR, flow cytometry, Western blotting, and Lionheart live-cell microscopy.

Results:
Mice fed H/L had significantly increased splenic T cell expression of phospho-STING, alongside decreased T cell
Xbp1
expression and increased cardiac inflammation, compared to STD-fed mice. Ex vivo incubation of CD4
+
T cells with PALNA significantly decreased
Xbp1
gene expression and increased apoptosis, indicated by cleaved caspase-3 and cleaved poly (ADP-ribose) polymerase (PARP) expression, compared to cells in CM. However, the remaining live PALNA-treated cells showed higher expression of the activation protein CD69, and this was sustained over time, compared to cells in CM. Strikingly, live cell imaging revealed that STING
-/-
T cells had improved survival and decreased CD69 expression in response to PALNA compared to WT PALNA-treated cells.

Conclusions:
STING contributes to T cell inflammation in experimental cardiometabolic HFpEF by synergizing with dysregulated ER stress responses to promote the emergence of hyperactivated T cells.
Abstract We081: β1 integrins regulate cellular behavior and cardiomyocyte organization during ventricular wall formation
Authors: Miao Lianjie , Lu Yangyang , Nusrat Anika , Zhao Luqi , Castillo Micah , Xiao Yongqi , guo Hongyan , Liu Yu , Gunaratne Preethi , Schwartz Robert , R Burns Alan , Kumar Ashok , DiPersio C. Michael , Wu Mingfu
Published: October 2024

Introduction:
Cardiomyocytes are surrounded by extracellular matrix (ECM) and interact with ECM via integrins. However, the mechanisms through which these cells organize themselves to establish the tissue architecture of trabeculae and the ventricular wall remain inadequately elucidated. Integrins are the primary cell surface receptors for adhesion to the ECM and mediate both inside-out and outside-in signal transduction pathways that control various cell functions, including proliferation, survival, migration, and gene expression. We found that the β1 integrin subunit (β1), encoded by
Itgb1
, is highly expressed in all cardiac cell types.

Hypothesis:
β1 integrins play a crucial role in regulating cardiomyocyte behavior and organization during ventricular wall morphogenesis in mice.

Methods and Results:
We applied mRNA deep sequencing and immunostaining to determine the expression repertoires of α/β integrins and their ligands in the embryonic heart. β1 and some of its ECM ligands are asymmetrically distributed and enriched in the luminal side of cardiomyocytes, and fibronectin surrounds cardiomyocytes, creating a network for them.
Itgb1
, which encodes the β1, was deleted via

Nkx2.5
Cre/+

to generate myocardial-specific
Itgb1
knockout (B1KO) mice. B1KO hearts lack a trabecular zone but a thicker compact zone. The levels of hyaluronic acid and versican, essential for trabecular initiation, were not significantly different between control and B1KO. Instead, fibronectin, a ligand of β1, was absent in the myocardium of B1KO hearts. Furthermore, B1KO cardiomyocytes display a random cellular orientation and fail to undergo perpendicular cell division, be organized properly, and establish the proper tissue architecture to form trabeculae. Mosaic clonal lineage tracing showed that
Itgb1
regulates cardiomyocyte transmural migration and proliferation autonomously.

Conclusions:
β1 is asymmetrically localized in the cardiomyocytes, and some of its ECM ligands are enriched along the luminal side of the myocardium, and fibronectin surrounds cardiomyocytes. β1 integrins are required for cardiomyocytes to attach to the ECM network. This engagement provides structural support for cardiomyocytes to maintain shape, undergo perpendicular division, and establish cellular organization. Deletion of
Itgb1
leads to loss of β1 and fibronectin and prevents cardiomyocytes from engaging the ECM network, resulting in failure to establish tissue architecture to form trabeculae.
Abstract We146: CHD8 is Essential for Postnatal Heart Function through Preserving Mitochondrial Integrity
Authors: Chen Nong , Sakabe Masahide , Xin Mei
Published: October 2024

Objective:
Epigenetic regulation plays a crucial role in heart development and disease, although its mechanisms remain elusive. Known mutations in epigenetic regulators have been linked to cardiac disease. For instance, the mutation in the chromodomain helicase DNA-binding (CHD) protein 7 gene, which leads to CHARGE syndrome, is known to results in heart defects. However, the role of CHD8, an interacting partner of CHD7, in the heart has not been investigated. This study aims to explore the function of CHD8 in the heart.

Methods and Results:
We generated a cardiac-specific Chd8 conditional knockout mouse line (Chd8 cKO) by breeding Chd8
fl/fl
mice with aMHC Cre mice. Over 60% of Chd8 cKO mice died between 7 and 9 months of age, with increased heart weight to bodyweight ratio and reduced cardiac function, increased fibrosis, and signs of dilated cardiomyopathy. Electron microscopy analysis revealed disordered sarcomeres and escalated mitophagy in CHD8-deficient hearts.

Using Genome-wide transcriptome analysis though RNA sequencing, we discovered that genes involved in mitochondrial oxidative phosphorylation and muscle contraction were down-regulated in Chd8 cKO hearts. Proteins in the electron transport chain complexes were down regulated and mitophagy marker LC3 increased in the Chd8 cKO hearts compared with the control mice. Upregulated pathways include inflammatory response suggesting that the innate immune response may contribute to dilated cardiomyopathy.

Conclusions:
Our findings underscore CHD8 as a key regulator of cardiac function and regeneration by influencing chromatin accessibility of genes involved in mitochondrial oxidative phosphorylation and contractility. Overall, our study provides new insights into the epigenetic regulation of cardiac function and the CHD8’s role in heart development and disease. It implies that activating CHD8 may be a promising therapeutic strategy for treating dilated cardiomyopathy and other heart-related diseases.
When Is a Menorah a “Menorah”? Text and Realia from Asia Minor
Authors: Wilson Mark
Published: October 2024

Menorahs found at sites in Asia Minor have been a primary indicator of the presence of Jewish communities in its cities. Menorahs were found in all three of the structures identified as synagogues in Turkey. This article addresses the following question: When is a menorah a “menorah”? It first discusses a translation conundrum related to a well-known text addressed to Jesus’ followers in seven cities in western Asia Minor by John, the author of Revelation. It then provides an extended discussion regarding menorahs and symbols identified as menorahs at sites in Rough Cilicia in southeastern Asia Minor. Finally, it assesses each one so as to determine whether it could properly considered a menorah.
Digital Rights in Palestine: Prospects of a Constitutional Judicial Review
Authors: Qandeel Mais
Published: October 2024

A set of rights and freedoms are protected by constitutions and interpreted by the competent judiciary. The increasing use of information and communication technologies (ICT s) creates new legal challenges to rights and their judicial interpretation. This article aims at analysing the prospects of the Palestinian Supreme Constitutional Court (SCC) concerning digital rights, while considering political and legal challenges. In doing so, it begins with a brief introduction to the concepts of digital right and digital constitutionalism. The article further describes the status of digital rights in Palestine and evaluates the role of the SCC in the adjudication of the constitutionality of the laws in relation to human rights protection on the internet. It finally offers proposals for understanding the necessity for constitutional protection of digital rights in Palestine through judicial review. It argues that the Palestine needs constitutional modernisation to address the legal implications of technologies on human rights.
Contributions of Ottoman Fatwā Collections to Inheritance Law: An Analysis of Legal Opinions by the 16th-Century Chief Muftīs
Authors: Saǧlam Hüseyin , Toprak M. Bedrettin
Published: October 2024

This article investigates the contributions of Ottoman fatwā collections to inheritance law, focusing on the 16th-century chief muftīs’ fatāwā. It emphasizes the significance of fatawa in understanding Ottoman inheritance law and explores various aspects of the division of estate, highlighting the socio-legal dynamics and the use of fatawa by individuals seeking legal advice. Within this framework, it examines why individuals preferred obtaining fatawa from chief muftīs for the division of the estate over other options. The methodology of this study involved a comprehensive analysis of fatwā collections from 16th-century Ottoman chief muftīs, focusing on categorizing and interpreting these fatawa to understand their impact on inheritance law. The study reveals that economic considerations, complex family structures, mathematical issues in estate division, and specific legal situations significantly influenced individuals’ preference for seeking fatāwā. Through analyzing fatwā collections, the study uncovers economic motivations behind this preference, revealing how fatāwā offered a cost-effective solution for heirs, alongside addressing specific legal challenges like ‘awl and munāsakha.
The nomenclatural status of “Anoplophallus maculatus” Cope, 1895, its consequences on the systematics of Lycodon subcinctus auctorum, and the description of a new species (Squamata: Colubridae)
Authors: NGUYEN TAN VAN , LEE JUSTIN L. , PAUWELS OLIVIER S. G. , KENNEDY-GOLD STEVIE R. , POYARKOV NIKOLAY A. , DAVID PATRICK , VOGEL GERNOT
Published: October 2024

The Malayan banded wolf snake, Lycodon subcinctus (F. Boie, 1827) has a complicated taxonomic and nomenclatural history. For almost a century, the combination Anoplophallus maculatus Cope, 1895 has been applied to populations of L. subcinctus from southern China, with various authors recognizing the taxon maculatus as a subspecies (i.e., Lycodon subcinctus maculatus) or junior synonym. A recent study that proposed the elevation of Chinese L. subcinctus to species rank (as Lycodon “maculatus”) prompted us to re-investigate several nomenclatural issues relevant to Anoplophallus maculatus that have not been fully addressed in the literature. We show that Anoplophallus maculatus was not described as a new species, and actually refers to Megalops maculatus Hallowell, 1861, a Neotropical snake currently known as Leptodeira maculata (Dipsadinae). As a consequence, populations of L. subcinctus recently recognized as Lycodon “maculatus”, have no valid binomen applied to them. To resolve this issue, we provide a new species name for populations of L. subcinctus from China and include a morphological description based on specimens collected across all of mainland Southeast Asia. The new species, Lycodon neomaculatus sp. nov., occurs well outside of southern China, and inhabits Indochina (Cambodia, Laos, Thailand, Vietnam) and eastern Myanmar. Lycodon neomaculatus sp. nov. is distinguished from both L. sealei and L. subcinctus by subtle differences in adult dorsal color pattern and body scalation. The distributions of L. neomaculatus sp. nov. and L. subcinctus sensu stricto appear to be allopatric, and the Isthmus of Kra region in peninsular Thailand may act as a biogeographic filter separating the two species.
Three new species of the genus Viettherchnus Kirejtshuk, 1985 (Coleoptera: Nitidulidae: Nitidulinae: Cyllodini)
Authors: JELÍNEK JOSEF , HÁJEK JIŘÍ
Published: October 2024

Three new species of the genus Viettherchnus Kirejtshuk, 1985 are described and illustrated: Viettherchnus apriculus sp. nov. (Malaysia), V. helferi sp. nov. (Indonesia: Kalimantan, Malaysia, Myanmar) and V. orszuliki sp. nov. (Nepal); the latter represents the first species of the genus known from the Himalayas. An identification key to all Viettherchnus species is provided.