bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 491: E-cigarette Aerosol Elevates Cardiovascular Oxidative Stress in Mice With Aldehyde Dehydrogenase 2 Deficiency
Authors: Yu Xuan , Zeng Xiaocong , Chen Ri , Sinharoy Pritam , Gross Eric R
Published: October 2020

Introduction:
E-cigarette aerosol contains reactive aldehydes including acetaldehyde, formaldehyde, and acrolein when e-cigarette is heated. Approximately 560 million people worldwide cannot efficiently metabolize aldehydes present in e-cigarette aerosol, due to a genetic deficiency in aldehyde dehydrogenase 2 enzyme (ALDH2), known as ALDH2*2. Little is known how aldehyde exposure from e-cigarettes, coupled with genetic differences in aldehyde metabolism, affects cardiovascular oxidative stress both at a physiological and cellular level.

Hypothesis:
E-cigarette aerosol exposure will elevate heart rate and cellular oxidative stress more substantially in ALDH2*2 knock-in mice versus wild type ALDH2 mice.

Methods:
To measure aldehyde levels, e-cigarette Juul aerosols were collected and quantified by selective ion flow gas mass spectrometry. Further, age-matched male wild type and homozygous ALDH2*2 mice (8-10 weeks old, ~25g) were implanted with EKG telemeters. After surgical recovery, mice were paired by genotype (one wild type ALDH2 and one ALDH2*2 mice) and exposed to either Juul aerosol or room air 4 sessions per day for 10 days. For each session, 7 puffs/min were drawn for the first two minutes (a total of 14 puffs), and the whole body exposure to Juul aerosol lasted 7 minutes, continued with 23 minutes smoking-free intervals in each session. Mice EKG waveforms were recorded daily. After 10 days of exposure, heart homogenates were subjected to biochemical assays including lipid peroxidation, 4-HNE protein adduct formation, and protein carbonylation.

Results:
Quantification of reactive aldehyde levels in e-cigarettes revealed that Juul aerosol contained acetaldehyde (5.3±0.32 ppm), formaldehyde (0.20±0.02 ppm), and acrolein (0.09±0.01 ppm). When exposed to Juul aerosol, ALDH2*2 mice showed a maximal increase in heart rate unlike ALDH2 wild type mice (774.6±29.5 bpm versus 678.9±32.8 bpm respectively, *
p
<0.01, n=8) at day 6. Furthermore, heart homogenates from ALDH2*2 mice demonstrated exacerbated oxidative stress, including higher level of 4-HNE adducts (1.5-fold), protein carbonyls (1.5-fold) and lipid peroxides (2-fold) relative to hearts from wild type ALDH2 mice, when both genotype mice were exposed to Juul e-cigarette aerosol (n=4/group).

Conclusions:
These findings indicate e-cigarette aerosols contain reactive aldehydes, primarily acetaldehyde. A deficiency in reactive aldehyde metabolism by having an ALDH2*2 deficiency may contribute to increases in heart rate and oxidative stress within the cardiovascular system while smoking e-cigarettes.
Abstract 495: A Reappraisal of Body Mass Index and Aerobic Fitness in Young Athletic Women
Authors: Kuznetsov Alexandra , Trager Lena , Loomer Garrett , Churchill Timothy , Weiner Rory B , Wasfy Meagan M , Guseh James S , BAGGISH Aaron L
Published: October 2020

Background:
Lower body mass is thought to enhance athletic performance. We explored the relationship between BMI and aerobic fitness in athletic younger and older men and women.

Methods:
We examined 2224 patients (805 women, 1419 men) ages 17 through 91, referred to the Cardiovascular Performance Program at Massachusetts General Hospital between 2011 and 2019. Cardiorespiratory fitness (CRF; Peak V0
2
) was assessed and expressed as %- predicted peak V0
2
, using the equation of Jones to adjust for age, height, and weight. Associations were evaluated using linear regression.

Results:
Mean peak VO
2
was 34.3±14.1 mL/kg/min, and median %-predicted VO
2
was 105%. We observed an inverse association between BMI and CRF in adults over 30 years old (
B
= -0.043, p = 3.3x10
-33
for men,
B
= -0.051, p = 1.6 x 10
-13
for women) (Fig 1a) and men under 30 (
B
= -0.025, p = 0.005) (Fig 1b). No linear association was found in younger women ages 17 - 29 (
B
= -0.0078, p = 0.38) (Fig 1b). In contrast, a quadratic model disclosed a direct relationship at lower BMI and an inverse relationship at higher BMI (Fig 1c). An examination of high fitness young adults (defined as having %-predicted VO
2
≥ 120%) demonstrates that fit women have mean BMI’s of 23.15 ± 2.69 (Fig 1d).

Conclusion:
These data highlight that BMI and fitness in younger aerobically fit women do not follow a simple linear relationship but one that is parabolic in nature. High fitness in young women occurs with highest density in the upper strata of the WHO and NIH-defined range of healthy BMI. These data support a deemphasis of weight loss generally and BMI specifically to enhance aerobic performance in athletic young women.
Abstract 498: Krueppel-like Factor 15 Modulation: A Crispr-based Re-establishment of Homeostatic Transcription in Diseased Mouse and Human Cardiomyocytes
Authors: Schoger Eric , Iyer Lavanya M , Noack Claudia , Rocha Cheila , Jassyk Mareike , Shomroni Orr , Salinas Gabriela , Zelarayan Laura C
Published: October 2020

An altered transcriptomic profile of cardiomyocytes is a hallmark of cardiac remodelling and heart failure. Re-activation of gene programs necessary for myocardial homeostasis and inhibition of deleterious signalling pathways, such as Wnt/beta-catenin signalling, is therefore therapeutically desirable, however challenging to achieve tissue-specifically. We identified Krueppel-like factor 15 (KLF15) as a cardiac specific inhibitor of Wnt/beta-catenin signalling, lost during heart failure progression in the mouse and human heart. To re-establish endogenous
Klf15
expression, we designed a cardiomyocyte specific CRISPR activation mouse model expressing enzymatically inactive Cas9 (dCas9) fused to a transcriptional activator (VPR) under
Myh6
promoter control. The dCas9VPR protein is directed by guide RNAs (gRNA), delivered by AAV9, to the promoter region of
Klf15
and induces gene expression. We tested activation of
Klf15
in the adult myocardium by injecting 15 weeks old Myh6-dCas9VPR mice systemically with AAV9 carrying
Klf15
gRNAs and increasing
Klf15
transcript levels 8 weeks later up to 1.6 fold compared to controls. Since we found a 0.5 fold change of
Klf15
expression upon transverse aortic constriction compared to sham operated mice, we expect the CRISPR activation induced
Klf15
activation to be sufficient to re-establish
Klf15
expression. As a translational approach and since we demonstrated KLF15-WNT regulation in human cardiomyocytes, we integrated dCas9VPR into human induced pluripotent stem cells (hIPSC) by targeted genome editing at the AAVS1 safe harbour locus and confirmed dCas9VPR expression in hIPSC and in hIPSC-derived cardiomyocytes (hIPSC-CM). We tested KLF15 gRNAs in HEK293 cells and selected the most efficient candidate gRNAs to test
Klf15
activation in dCas9VPR-IPSC resulting in an up to 1.7 fold activation compared to controls. For gene activation in dCas9VPR hIPSC-CM, we are currently testing lentivirus constructs to deliver up to 3 gRNAs simultaneously to tailor KLF15 expression. With these prerequisites, we aim to re-activate master regulators of normal cardiomyocyte function in failing cardiomyocytes at physiologically relevant levels, such as KLF15, to test potential therapeutic targets for the prevention of heart failure progression.
Abstract 501: Hydrogen Sulfide Protects the Heart Against Ferroptotic Cell Death in Diabetic Cardiomyopathy
Authors: Kar Sumit , Shahshahan Hamid , Mishra Paras K
Published: October 2020

Hyperglycemia-induced death of terminally differentiated cardiomyocytes in diabetic cardiomyopathy (DMCM) leads to heart failure. Ferroptosis is a newly discovered form of cell death triggered by intracellular iron and oxidative stress. While little is known about ferroptosis in DMCM, hyperinsulinemia stimulates intracellular iron uptake, which would be predicted to increase ferroptosis. H
2
S is an endogenous gaseous signaling molecule which may inhibit ferroptosis. H
2
S donors increase glutathione substrate for glutathione peroxidase 4 (GPX4), which removes lipid peroxidation and is the primary inhibitor of ferroptosis. However, no study has investigated the role of H
2
S in ferroptosis. We tested the hypothesis that increased ferroptosis contributes to DMCM, which is ameliorated by restoring H
2
S levels, by measuring ferroptosis in the hearts of db/db mice with DMCM and high glucose treated primary neonatal cardiomyocytes after H
2
S donor treatment. GPX4 expression was decreased (3.21±0.41 GPX4/total protein in db/+ control mice vs. 1.84±0.26 in db/db, P<0.05, n=6/group) and 4-HNE lipid peroxide was increased (7.37±0.98 μg 4-HNE/total protein in db/+ control mice vs. 8.73±0.84 in db/db, P<0.05, n=3/group) in the left ventricle of db/db mice indicating increased ferroptosis in DMCM. GPX4 activity was also decreased along with increased 4-HNE in high glucose cultured cardiomyocytes. Treatment with the ferroptosis inhibitor ferrostatin-1 prevented hyperglycemia induced ferroptosis. Treatment of cardiomyocytes with the H
2
S donor GYY4137 in hyperglycemia also decreased 4-HNE. We also validated the anti-ferroptotic potential of H
2
S by treating cardiomyocytes with the ferroptosis inducer RSL3 and GYY4137. H
2
S donor treatment reduced reactive oxygen species and 4-HNE lipid peroxide seen after ferroptosis induction with RSL3. This study establishes ferroptosis as a new, non-apoptotic, form of cell death in DMCM, and H
2
S as a novel regulator of cardiac ferroptosis.
Abstract 531: Mechanistic and Functional Differences in Rna Binding and Processing Activities of the Muscle- and Non-muscle Rbfox2 Isoforms
Authors: Misra Chaitali , Valiya Chembazhi Ullas , Matatov Sarah , Bangru Sushant , Kalsotra Auinash
Published: October 2020

Myotonic Dystrophy type 1 (DM1), the most prevalent form of adult onset muscular dystrophy, is caused by CTG trinucleotide repeat expansion in the 3’-UTR of DMPK gene. Over 80% of DM1 patients exhibit heart dysfunctions, which are the second leading cause for DM1-related deaths. Recently, we demonstrated that aberrant expression of a non-muscle splice isoform of RNA-binding protein RBFOX2 triggers cardiac conduction delay, atrioventricular heart blocks, and spontaneous arrhythmogenesis in DM1 heart. RBFOX2 is a master regulator of tissue-specific alternative splicing and a pair of mutually exclusive 43-nucleotide(nt) and 40-nt exons in its C-terminal domain encode the muscle (RBFOX2
43
) and non-muscle (RBFOX2
40
) isoforms. The RBFOX2
40
isoform is predominantly expressed in the fetal heart, and is replaced by the RBFOX2
43
isoform in development, specifically within the cardiomyocytes of adult hearts. To deconstruct the splicing regulatory networks of RBFOX2
43
and RBFOX2
40
isoforms, characterize their respective RNA binding landscapes, and determine the RBFOX2
40
-driven transcriptome alterations in DM1 heart tissue, we performed eCLIP and high-resolution RNA-sequencing studies on cardiomyocytes isolated from wild type (expressing the normal muscle-specific RBFOX2
43
isoform),
Rbfox2
Δ43/Δ43
(expressing the non-muscle RBFOX2
40
isoform), and RBFOX2
40
overexpressing (OE) mice. By integrating genome-wide RNA binding and processing activities for the two RBFOX2 isoforms, we found that a switch from the muscle-specific (RBFOX2
43
) to non-muscle (RBFOX2
40
) isoform provokes DM1-like cardiac pathology by altering the mRNA abundance and splicing of genes encoding components of the conduction system and/or contractile apparatus. Further, through subnuclear fractionation and protein-protein interaction studies, we demonstrate that the higher-order assembly of LASR (large assembly of splicing regulators) complexes formed by the RBFOX2
40
isoform boost its splicing activity and promote the generation of pathogenic splice variants of voltage-gated ion channels and other components of the cardiac conduction system.
Abstract 541: A Hedgehog-signaling Dependent Gene Regulatory Network Directly Delays Second Heart Field Differentiation to Prevent Congenital Heart Disease
Authors: Rowton Megan , Rydeen Ariel , Carlos Perez-Cervantes , Deng Nikita , Lu Emery , Jacobs-Li Jessica , Hoffmann Andrew , Steimle Jeffrey , Yang Xinan Holly , Guzzetta Alexander , Lazarevic Sonja , Kim Chul , Yu Shuhan , Koska Mervenaz , Hanson Erika , Chan Sunny Sun-Kin , Garry Daniel J , Ikegami Kohta , Kyba Michael , Moskowitz Ivan P
Published: October 2020

The first heart field (FHF) and the second heart field (SHF) comprise the major progenitor pools for the developing vertebrate heart. We investigated the delayed differentiation of the SHF relative to the FHF, a major distinguishing feature of these fields. Single-cell transcriptional profiling of the SHF revealed a differentiation trajectory of SHF progenitors to cardiomyocytes. Hedgehog (Hh) signaling was highly enriched in the progenitor state in signaling pathway analysis, suggesting a possible role in cardiomyocyte differentiation control. Transcriptional profiling of the SHF following removal of active Hh signaling
in vivo
revealed inappropriate cardiomyocyte gene expression. We observed precocious cardiomyocyte differentiation in the SHF
in vivo
in Hh mutants, which led to Congenital Heart Disease (CHD). Modeling active Hh signaling in a novel mouse embryonic stem cell (mESC) line through transient expression of the activating Hh transcription factor (TF), GLI1, in cardiac progenitors delayed the onset of cardiomyocyte differentiation. GLI1 directly activated a progenitor-specific gene regulatory network, dominated by repressive TFs, that prevented the acquisition of the cardiomyocyte gene regulatory network. Maintained expression of one GLI1 target TF, FOXF1, repressed the cardiomyocyte differentiation program. FOXF1 binding sites were identified at putative regulatory elements near repressed cardiac genes involved in contraction, electrical impulse propagation and transcriptional regulation. Finally, FOXF1 repressed the activity of these elements
in vitro
, indicating that FOXF1 can directly repress the activation of genes essential for cardiomyocyte differentiation. Together, these results indicate that a Hh-dependent gene regulatory network including transcriptional repressors directly delays the onset of cardiomyocyte gene expression to delay SHF differentiation. Abrogation of Hh signaling and the resultant premature differentiation of cardiac progenitors provides a molecular mechanism for the ontogeny of some CHD.
Abstract 552: Bradykinin Receptor B1/ Matrix Metalloprotease 3 Pathway is a Novel Target in Preventing Cardiac Ischemia-reperfusion Injury
Authors: Zhang Qin , Ai Lizhuo , Liu Lifeng , Betancourt Cristian , Knapp Maura , Sitikov Albert , Sharp Willard W , Liao James K , Wu Rongxue
Published: October 2020

Introduction:
Impaired endothelial function leads to the progression of heart failure after Ischemia-reperfusion (IR). Kinin activation of bradykinin receptor 1 (B1R), a G protein-coupled receptor that has been found to induce capillary leakage, may serve as a critical mediator in cardiac microvascular barrier dysfunction. However, the underlying mechanisms are not clear. We found that B1R inhibition abolished IR-induced endothelial matrix metalloprotease (MMP3) expression and improved endothelial barrier formation. Thus, we hypothesized that B1R antagonist protects against cardiac IR injury through an MMP3 pathway.

Methods and Results:
MMP3-/- mice and their littermate controls (WT) were subjected to either cardiac IR or sham control. The baseline characteristics of these mice showed minimal phenotypes. Cardiac function was determined at 3, 7 and 24 days post-IR by echocardiography. The MMP3-/- mice displayed improved cardiac function compared to the control mice, as determined by fractional shortening (26% ± 1.1 MMP3-/- vs. 21% ± 0.9 WT, p<0.05, n=5) and ejection fraction (48% ± 1.9 MMP3-/- vs. 42% ± 2.8.1 WT, p<0.05, n=5), and treating with B1R antagonist (300 μg/Kg) significantly reduced serum MMP3 levels (p<0.01). Compared to the control mice, MMP3-/- mice had significantly less infarction/area at risk 24 hours post-IR demonstrated through TTC staining. In vitro studies revealed that cellular hypoxia-reoxygenation (HR) injury significantly increased both B1R and MMP3 expression levels in primary isolated cardiac mice microvascular endothelial cells (mCMVEC). MMP3 levels were measured via ELISA. Moreover, B1R agonist treatment (1uM) increased MMP3 levels, while the use of the antagonist attenuated the increase of MMP3 in response to HR. Finally, the use of B1R antagonist improved MMP3 induced endothelial barrier dysfunction, which was measured by the electric cell-substrate impedance sensing (ECIS) system. Taken together, the results demonstrated that B1R antagonist abolished IR induced MMP3 induction and that the deletion of MMP3 is protective of cardiac function upon IR injury.

Conclusions:
MMP3 is a critical regulator of cardiac microvascular barrier function, and B1R/MMP3 could potentially serve as a novel therapeutic target for heart failure in response to IR injury.
Abstract MP120: Ryanodine Receptor 2 Prevents Endoplasmic Reticulum Stress-induced Defects in Cardiomyocyte Maturation
Authors: Guo Yuxuan , Jardin Blake , Guatimosim Silvia , Pu William T
Published: October 2020

Cardiomyocyte (CM) maturation is an essential step in heart development that prepares the organ for robust and sustainable contractions. However, very little is known about the contribution of CM maturation to cardiac pathogenesis, partly due to the paucity of basic knowledge about maturation. For example, although calcium plays well-established roles in cardiac stress responses, whether normal calcium oscillations are required for CM maturation has not been examined. To address this question, we created mice with mosaic inactivation of ryanodine receptor 2 (RYR2), the central channel that mediates calcium release from sarcoplasmic/endoplasmic reticulum (SR/ER), in CMs. RYR2 was depleted in a small fraction of neonatal CMs by adeno-associated virus (AAV)-mediated Cas9-based somatic mutagenesis (CASAAV). The majority of CMs intentionally retained RYR2 expression, which circumvented cardiac dysfunction and death that would result from widespread calcium transient ablation. In RYR2-depleted CMs, sarcomere expansion, transverse-tubule formation, mitochondrial metabolism and maturational hypertrophy were impaired. Strikingly, these maturation defects were associated with unfolded protein response, with the activation of three major ER stress effectors: spliced Xbp1 (XBP1s), ATF6 N-terminus (ATF6N), and ATF4. Tauroursodeoxycholic acid, an ER stress-relieving drug, partially rescued the maturation defects in RYR2-depleted CMs. Over expression (OE) of XBP1s, ATF6N or ATF4 each impaired CM maturation, with phenotypic severity ranked as ATF4>XBP1s>ATF6N. ATF4 OE in neonatal CMs triggered both ultrastructural and transcriptomic changes highly correlated with RYR2 depletion phenotypes. Interestingly, ATF4 OE altered distinct transcriptomic programs in neonatal vs. adult CMs and caused ultrastructural defects only at the neonatal stage. No terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was detected upon RYR2 depletion or ATF4 OE, excluding the contribution of apoptosis to maturation defects. Collectively, these data indicate that RYR2-mediated calcium handling prevents an adverse and stage-specific impact of ER stress on CM maturation, which lead to CM dysfunction.
Abstract MP152: Smad1 Activation in Infarct Macrophages Restrains Angiogenesis and Accentuates Adverse Remodeling of the Infarcted Myocardium
Authors: Chen Bijun , Su Ya , Frangogiannis Nikolaos G
Published: October 2020

Macrophages play multiple roles in repair and remodeling of the infarcted heart, contributing to phagocytosis of dead cells, regulation of inflammation, fibrosis and angiogenesis. TGF-β superfamily members are critically involved in regulation of macrophage phenotype through the Smad cascades. In most cell types, TGF-βs signal predominantly by activating Smad2/3 signaling, whereas BMPs act through Smad1/5-mediated pathways. We previously showed that TGF-β/Smad3 signaling in macrophages protects the infarcted heart from adverse remodeling by mediating phagocytotic activity and anti-inflammatory transition. However, the in vivo role of Smad1/5 signaling in macrophages remains unknown. We examined the role of macrophage-specific Smad1 signaling in a mouse model of myocardial infarction (MI). Smad1 is activated in a subset of myeloid cells infiltrating the infarcted myocardium. In vitro, TGF-β1, TGF-β3, BMP4, BMP7, but not TGF-β2, BMP2, BMP6 markedly activated Smad1/5 signaling in macrophages. In order to examine the role of Smad1 in regulation of macrophage phenotype we generated myeloid cell-specific Smad1 KO mice (MyS1KO). MyS1KO mice had better survival compared to S1fl/fl controls 28 days post MI (
p
=0.0009, n=26-27). Echocardiographic data showed that MyS1KO mice had reduced left ventricular dilation and attenuated systolic dysfunction 28 days after MI. Although scar size was comparable between groups at the 7 day timepoint, MyS1KO animals had significantly smaller scars 28 days after MI, suggesting improved scar remodeling. MyS1KO mice exhibited increased microvessel density in the infarct, suggesting that Smad1 activation in macrophages may restrain angiogenesis. In order to explore the mechanisms, we isolated infarct macrophages from MyS1KO and S1fl/fl hearts and compared angiogenesis-related protein expression by performing a proteomic array. Smad1 loss in macrophages did not affect VEGF-A expression, but significantly increased levels of the angiogenic chemokine CXCL12. Our findings suggest that Smad1 is directly activated by TGF-βs in macrophages, and that Smad1 activation in infarct macrophages may promote adverse remodeling by restraining angiogenic effects through suppression of angiogenic chemokines.
Abstract MP154: Deletion of the Duffy Antigen Receptor for Chemokines (DARC) Confers Protection Against Abdominal Aortic Aneurysm (AAA) Formation
Authors: Benson Tyler W , Horimatsu Tetsuo , Ogbi Mourad , Kim Ha Won W , Weintraub Neal L
Published: October 2020

Introduction:
Abdominal aortic aneurysms (AAA) are characterized by inflammation and matrix metalloproteinase (MMP)-mediated degradation of extra cellular matrix proteins leading to aortic dilation and potentially rupture. Inflammatory chemokines that promote AAA are modulated by binding to the Duffy antigen receptor for chemokines (DARC), a non-signaling receptor expressed primarily on erythrocytes. Interestingly, African Americans exhibit reduced frequency of AAA, and the majority of African descendent individuals do not express DARC on their erythrocytes. Here, we tested the hypothesis that DARC gene deletion protects against the development of AAA.

Methods:
The induction of AAA was performed using both angiotensin II (AngII) infusion and calcium chloride (CaCl
2
) application models. Eight week old LDLR knockout (KO, control) and LDLR KO/DARC KO mice were infused with AngII via osmotic mini-pumps for four weeks. For the CaCl
2
application model, twelve week old DARC KO and WT control mice underwent laparotomy and 0.5 mol/L CaCl
2
was applied to the infrarenal aorta. Aortic dilation and AAA formation was assessed using ultrasound, mice were then euthanized and tissues were harvested for analysis.

Results:
The aortic diameter of LDLR KO/DARC KO mice was significantly lower relative to control mice after AngII infusion (P=0.02). There was no difference in the pressor response to AngII between groups. Furthermore, IL-6 levels were significantly reduced in the aortas of LDLR KO/DARC KO mice compared to control mice (P=0.001). Expression of MCP-1, which has strong affinity for DARC, tended to be higher in aortas from LDLR KO/DARC KO mice compared to control (P=0.063). In contrast, plasma IL-6 levels were similar in both LDLR KO/DARC KO and control mice, while MCP-1 tended to be lower in the plasma of LDLR KO/DARC KO (P=0.07). DARC KO mice likewise exhibited a trend toward reduced aortic dilation in the CaCl
2
application model compared to controls (P=0.09).

Conclusions:
DARC KO mice are protected against AAA formation, perhaps through differential regulation of aortic chemokine trafficking. Understanding the mechanisms by which loss of DARC confers protection from AAA formation may be relevant to ethnic differences in susceptibility to AAA.
Abstract MP174: Cardiomyocyte-specific Deletion of Parathyroid Hormone Receptor 1 During Development Leads to Left Ventricular Hypoplasia
Authors: Leach Desiree , Jorgensen Ryan , Wolfe Jayne , Wilsbacher Lisa D
Published: October 2020

Congenital heart defects occur in approximately 1% of all births and signify the most common human birth defects. Our lab previously demonstrated that cardiomyocyte specific deletion of Sphingosine 1-Phosphate Receptor 1 (S1P
1
), a G protein-coupled receptor (GPCR) that couples exclusively to Gαi, leads to ventricular noncompaction in mice. Gαs- and Gαi-coupled GPCRs activate and inhibit adenylate cyclase, respectively, thereby regulating cAMP production. These GPCRs play important roles in adult cardiac health and disease, but their contributions to heart development are not well understood. We hypothesized that tight regulation of cAMP via Gαs- and Gαi-coupled GPCRs is a critical determinant of cardiomyocyte maturation and cardiac development. Here, we report that Parathyroid Hormone Receptor 1 (Pth1r) which couples to Gαs, is a mediator of cardiomyocyte maturation. Using mice with a Cre allele in the myosin light chain 2a locus (
Myl7
Cre/+,
also known as
Mlc2a
Cre/+
), we found that cardiomyocyte-specific deletion of Pth1r during development led to reduced postnatal survival. Hearts collected from surviving Pth1r cardiomyocyte mutant mice displayed left ventricular (LV) hypoplasia, and fetal cardiac genes Nppa (natriuretic peptide A) and β myosin heavy chain (Myh7) were upregulated in the hearts of surviving mutants. At 14.5 days post coitus (dpc) and 18.5 dpc, Pth1r cardiomyocyte mutant mice also showed LV hypoplasia. Furthermore, we observed decreased cardiomyocyte proliferation in Pth1r cardiomyocyte mutant hearts during development. Isolated cardiomyocytes from C57BL/6 wild-type embryos showed increased cAMP production after stimulation of Pth1r; Pth1r-dependent cAMP increases were inhibited by activation of S1P
1
. Overall, our results indicate a critical role for Pth1r signaling during heart development and suggest that cAMP levels must be tightly controlled in cardiomyocytes for normal cardiac development. LV hypoplasia is one of the key features of hypoplastic left heart syndrome (HLHS), one of the most severe forms of congenital heart disease; these studies may provide new approaches for the study and treatment of LV hypoplasia and HLHS.
Arguments Employed in Favour of Unjust Action in Euripides, Thucydides, and Plato
Authors: Hill Theodore
Published: October 2020

This article examines arguments used in the fifth century in favour of unjust action. Three main lines of argument are distinguished: (i) arguments based on ordinary human behaviour, employed by Glaucon in Plato’s Republic and by the Athenians in Thucydides’ Melian Dialogue, (ii) arguments based on self-interest, found also in Plato and Thucydides, and (iii) arguments based on setting against justice another value, normally courage (ἀνδρεία) or wisdom (σοφία). These arguments are absent in Hesiod and Aeschylus, but present later in Euripides, Thucydides and Plato. Categorising them allows us to understand more precisely a central intellectual phenomenon of the fifth century, and to identify how the argumentation, and therefore the discourse of power, employed in these sources varies according to genre. In particular, Euripides, in his portrayal of individuals, deals with these issues differently, and employs different arguments, from Thucydides, who is concerned with the conduct of poleis.
Roman Law in the Curriculum of the First Chinese Students in England, France, and China
Authors: Chen Li
Published: October 2020

This article retraces the beginnings of Roman law studies by Chinese students during the latter part of the 19th century. It relies on archival research in order to piece together the curricula and careers of three pioneering Chinese law students who first came to study law, including Roman law, in England, France, and China. Wu Tingfang’s legal training at an Inn of Court in London, Ma Kié-Tchong’s legal education at the University of Paris and Wang Chung Hui’s study at Peiyang University in Tianjin, all included a more or less in-depth exposure to Roman law. Ma Kié-Tchong’s wrote a thesis on Roman law in Latin. As the first surviving specimen of legal Latin written by a Chinese jurist, his work not only reflects Roman law studies in France in the 19th century, it also sheds light on the level of proficiency in legal Latin which a Chinese scholar could attain.
Les minorités dans la Déclaration Universelle des Droits de l’Homme, une absence délibérée
Authors: Arlettaz Fernando
Published: October 2020

The League of Nations established, in the interwar period, a legal regime for the protection of minorities which considered them as intermeditate groups between the State and the individuals. On the contrary, the Universal Declaration on Human Rights, adopted in 1948 by the United Nations, assumed a radically individualistic point of view and did not include any mention to minority rights. The travaux préparatoires of the Universal Declaration suggest that the question of minorities caused strong tension among States and that, for this reason, they avoided its inclusion in the 1948 document.
Psychodynamics of the Technological Unconscious
Authors: Ashraf Cameran
Published: October 2020

The shadow of technological development has received significant attention. User tracking, emotional manipulation, disinformation, online radicalization, and restrictions on free speech have shattered the cyber-utopianism of the 1990s and early 2000s. Yet the solutions to address technology’s problems are framed as requiring more technology. This technological solutionism and its subsequent choices mask unconscious processes which give rise to new technologies. This essay is an attempt to interrogate the history of these choices by taking a depth psychological approach to the technological unconscious, beginning with the impact of writing on the psyche to the advent of computer screens. With technologies becoming more sophisticated and a primary way we interact with reality, it is vital to create a body of depth psychology literature on technology’s history and impacts.
A new Ephydridae (Diptera) from the East Frisian Island of Borkum (Wadden Sea, Germany)
Authors: Stuke Jens-Hermann
Published: October 2020

Hyadina borkumensis
sp. n. is described from the East Frisian Island of Borkum (Germany, Lower Saxony, Wadden Sea) based on a single female specimen. The new species is characterized by the basally bent, brown wings with large white spots in radial cells r2+3, r4+5 and apical of crossvein dm-cu and its short radial vein R2+3. The new species shares the lateroclinate fronto-orbital seta with representatives of the Australasian–Oceanian genus Parahyadina Tonnoir & Malloch, 1926. The implication of this shared character for the use of relevant Ephydridae keys is briefly discussed.
Is Religious Intolerance Good for Your Health?
Authors: Burke Ciarán
Published: October 2020

Legal responses to the covid-19 pandemic have varied widely. Korea represents an interesting case study, as it seemed particularly well prepared, having enacted legislation in the wake of the mers outbreak, in 2015, to tackle future pandemics. This obviated recourse to emergency powers legislation, and couched Korea’s response in normal legislation, which tends to raise fewer human rights concerns than may arise under emergency measures. Despite this, however, Korea’s response to covid-19 raises significant questions about its compliance with core human rights norms under the International Convention on Civil and Political Rights, including freedom of religion and non-discrimination. These arose with regard to the state’s treatmennt of members of the Shincheonji Church of Jesus (scj), a relatively small, occasionally controversial, religious group. The treatment of the scj by the Korean state raises questions about whether its legal approach to tackling covid-19 was fit for purpose.
Research on the Motion Control Trajectory and Structure Design of a Lower Limb Standing Rehabilitation Training Device
Authors: Shi Yaochen , Lingyangmeng  , Chen Daimin , Chen Guoping , Zhao Zhongwei
Published: October 2020

Aiming at the problem of standing rehabilitation of lower limb motor dysfunction patient. Firstly, the standing movement process of human body is analyzed. Secondly, the exercise trajectory test and pressure distribution measurement test of healthy human body are studied. Thirdly, the
structure of the lower limb standing rehabilitation training device is designed according to the collected joint trajectory. Finally, the movement control trajectory simulation of the lower limb standing rehabilitation training device is carried out according to the collected regional pressure.
The simulation results of virtual prototype coincide with the test trajectory, which shows that the structure of the designed lower limb standing rehabilitation training device is reasonable, and the hybrid control method of force and position can realize the standing rehabilitation training
for lower limb motor dysfunction patient in the light of the set exercise trajectory.
The Construction of High-Performance Optoelectronic Interconnection Network and the Realization of Cloud Computing for Traffic Data Recognition
Authors: Zhang Shaofang , Li Xianjun , Wang Yuechun
Published: October 2020

Due to the problems of congestion, insufficient scalability, and high cost in the network core layer link, the network interconnection architecture needs to be optimized. The photoelectric interconnection technology has attracted the attention of many researchers with its high bandwidth,
low latency, and low power consumption. In this study, after fully considering the requirements of optoelectronic switching on network node degree and processing delay and corresponding to different transmission characteristics of circuit switching and optical circuit switching, an optoelectronic
interconnection architecture supporting heterogeneous network topology, namely THtop, is proposed. According to the optical circuit switching/circuit switching characteristics of the Internet, the corresponding topology structure is designed respectively in this architecture to enhance the
system integration and reduce the cost. At the same time, in order to meet the requirements of wavelength and number of hops of optical circuit switching, distributed control is designed in THtop, and the design method of heterogeneous optical switching unit is introduced in the switching
equipment, so that the optical switching node can further reduce the cost while playing its role. The use of high-speed optoelectronic networks will cause a large amount of traffic on the channel. In this study, the traffic identification is analyzed on the proposed THtop architecture. The
Map Reduce framework in cloud computing is used as the identification model, and the K-means algorithm is taken as the identification algorithm. According to its characteristics, the W_K-means algorithm is proposed for the type identification of traffic. The results show that
under the OpenNet Internet simulation environment, when the traffic is set to a certain level, more than 90% of the non-blocking network switching can be realized under the THtop architecture, and the completion time of the mouse flow and elephant flow can be significantly improved. Therefore,
under the Map Reduce model, the proposed W_K -means algorithm can quickly identify different traffic types.
Design of a New Type of Optical Fiber Infrasound Sensor and Its Usage in Karst Geological Exploration
Authors: Gong Fang
Published: October 2020

In this study, the demand of optical fiber (OF) sonic sensor is analyzed, and a composite OF infrasound sensor is proposed aiming at the detection demand of low-frequency infrasound waves in specific occasions. With this sensor, the polymer thin film and the aluminum soil can be combined
to form a kind of composite film, the ECF/P interferometer can be formed between the end faces of the ferrule of the OF FC contact, the sensor head of the OF ECF/P infrasound sensor system is fabricated and then packaged. The designed OF infrasound sensor is used in the construction of a wireless
sensor network (WSN) for karst geological exploration. The structure of the WSN includes the SimpliciTI communication network protocol, ADF7021 RF transceiver chip, STM32 processor chip, etc. New types of environment from karst can be sampled with this WSN. During the test, the performance
of the composite OF infrasound sensor is analyzed at the beginning. The acoustic sensitivity of up to –136.73 dB re 1 V/μPa can be obtained in the low frequency infrasound range (1–20 Hz); the WSN based on the composite OF infrasound sensor has lower power consumption
and faster networking speed. In addition, it can realize signal cover in karst areas without public mobile cellular networks, and send the information collected by the sensor to the data center, thus proving the fiber optic sound wave sensor designed in this study is successful and helpful
for the geological prospectors to monitor the environment.
First Person: Reporting for duty
Authors: Marshall Kim
Published: October 2020

Kim Marshall had not planned to become a teacher, but the specter of the Vietnam draft led him to seek a deferment by teaching in an urban school. He had no idea at the time that he was beginning a lifelong career. Here, he reflects on his struggle to manage his classroom and teach effectively that first year and the wake-up call that pushed him to improve for the sake of his students.
Rituximab in people living with HIV affected by immune-mediated renal diseases: a case-series
Authors: Alfano G , Giaroni F , Fontana F , Neri L , Mosconi G , Mussini C , Guaraldi G , Cappelli G
Published: October 2020

Over the last two decades, rituximab (RTX) has played an important role in the treatment of some lymphoproliferative malignancies and immune-mediated diseases. RTX administration is generally safe and well-tolerated, but side effects including late-onset neutropenia, hypogammaglobulinemia, hepatitis B reactivation and rare cases of progressive multifocal leukoencephalopathy have been observed after its administration. Although there are no absolute contraindications regarding its use in people living with HIV (PLWH), the prescription of this drug has been principally limited in patients with oncohematological diseases. In this report, we described the outcome of four PLWH who underwent RTX therapy after the diagnosis of immune-mediated renal disease. The main RTX-associated adverse effects were leukopenia, late-onset neutropenia and decline of CD4+ and CD8+ T-cell counts. In addition, two of the four patients experienced pneumonia requiring hospitalization within six months from the last RTX infusion. We suggest that RTX should be used with caution in PLWH until further evidence emerges on its safety profile in this vulnerable population.
Acute myeloid leukemia–induced remodeling of the human bone marrow niche predicts clinical outcome
Authors: Chen Yiyang , Hoffmeister Lina Marie , Zaun Yasmin , Arnold Lucas , Schmid Kurt Werner , Giebel Bernd , Klein-Hitpass Ludger , Hanenberg Helmut , Squire Anthony , Reinhardt H. Christian , Dührsen Ulrich , Bertram Stefanie , Hanoun Maher
Published: October 2020

Murine models of myeloid neoplasia show how leukemia infiltration alters the hematopoietic stem cell (HSC) niche to reinforce malignancy at the expense of healthy hematopoiesis. However, little is known about the bone marrow architecture in humans and its impact on clinical outcome. Here, we dissect the bone marrow niche in patients with acute myeloid leukemia (AML) at first diagnosis. We combined immunohistochemical stainings with global gene expression analyses from these AML patients and correlated them with clinical features. Mesenchymal stem and progenitor cells (MSPCs) lost quiescence and significantly expanded in the bone marrow of AML patients. Strikingly, their HSC- and niche-regulating capacities were impaired with significant inhibition of osteogenesis and bone formation in a cell contact–dependent manner through inhibition of cytoplasmic β-catenin. Assessment of bone metabolism by quantifying peripheral blood osteocalcin levels revealed 30% lower expression in AML patients at first diagnosis than in non-leukemic donors. Furthermore, patients with osteocalcin levels ≤11 ng/mL showed inferior overall survival with a 1-year survival rate of 38.7% whereas patients with higher osteocalcin levels reached a survival rate of 66.8%. These novel insights into the human AML bone marrow microenvironment help translate findings from preclinical models and detect new targets which might pave the way for niche-targeted therapies in AML patients.
Heterozygous germ line CSF3R variants as risk alleles for development of hematologic malignancies
Authors: Trottier Amy M. , Druhan Lawrence J. , Kraft Ira L. , Lance Amanda , Feurstein Simone , Helgeson Maria , Segal Jeremy P. , Das Soma , Avalos Belinda R. , Godley Lucy A.
Published: October 2020

Colony-stimulating factor 3 receptor (CSF3R) encodes the receptor for granulocyte colony-stimulating factor (G-CSF), a cytokine vital for granulocyte proliferation and differentiation. Acquired activating heterozygous variants in CSF3R are the main cause of chronic neutrophilic leukemia, a hyperproliferative disorder. In contrast, biallelic germ line hypomorphic variants in CSF3R are a rare cause of severe congenital neutropenia, a hypoproliferative condition. The impact of heterozygous germ line CSF3R variants, however, is unknown. We identified CSF3R as a new germ line hematologic malignancy predisposition gene through analysis of 832 next-generation sequencing tests conducted in 632 patients with hematologic malignancies. Among germ line CSF3R variants, 3 were abnormal in functional testing, indicating their deleterious nature. p.Trp547* was identified in 2 unrelated men with myelodysplastic syndromes diagnosed at 76 and 33 years of age, respectively. p.Trp547* is a loss-of-function nonsense variant in the extracellular domain that results in decreased CSF3R messenger RNA expression and abrogation of CSF3R surface expression and proliferative responses to G-CSF. p.Ala119Thr is a missense variant found in 2 patients with multiple myeloma and acute lymphoblastic leukemia, respectively. This variant is located between the extracellular immunoglobulin-like and cytokine receptor homology domains and results in decreased G-CSF sensitivity. p.Pro784Thr was identified in a 67-year-old man with multiple myeloma. p.Pro784Thr is a missense variant in the cytoplasmic domain that inhibits CSF3R internalization, producing a gain-of-function phenotype and G-CSF hypersensitivity. Our findings identify germ line heterozygous CSF3R variants as risk factors for development of myeloid and lymphoid malignancies.
Horizontal meta-analysis identifies common deregulated genes across AML subgroups providing a robust prognostic signature
Authors: Nehme Ali , Dakik Hassan , Picou Frédéric , Cheok Meyling , Preudhomme Claude , Dombret Hervé , Lambert Juliette , Gyan Emmanuel , Pigneux Arnaud , Récher Christian , Béné Marie C. , Gouilleux Fabrice , Zibara Kazem , Herault Olivier , Mazurier Frédéric
Published: October 2020

Advances in transcriptomics have improved our understanding of leukemic development and helped to enhance the stratification of patients. The tendency of transcriptomic studies to combine AML samples, regardless of cytogenetic abnormalities, could lead to bias in differential gene expression analysis because of the differential representation of AML subgroups. Hence, we performed a horizontal meta-analysis that integrated transcriptomic data on AML from multiple studies, to enrich the less frequent cytogenetic subgroups and to uncover common genes involved in the development of AML and response to therapy. A total of 28 Affymetrix microarray data sets containing 3940 AML samples were downloaded from the Gene Expression Omnibus database. After stringent quality control, transcriptomic data on 1534 samples from 11 data sets, covering 10 AML cytogenetically defined subgroups, were retained and merged with the data on 198 healthy bone marrow samples. Differentially expressed genes between each cytogenetic subgroup and normal samples were extracted, enabling the unbiased identification of 330 commonly deregulated genes (CODEGs), which showed enriched profiles of myeloid differentiation, leukemic stem cell status, and relapse. Most of these genes were downregulated, in accordance with DNA hypermethylation. CODEGs were then used to create a prognostic score based on the weighted sum of expression of 22 core genes (CODEG22). The score was validated with microarray data of 5 independent cohorts and by quantitative real time-polymerase chain reaction in a cohort of 142 samples. CODEG22-based stratification of patients, globally and into subpopulations of cytologically healthy and elderly individuals, may complement the European LeukemiaNet classification, for a more accurate prediction of AML outcomes.
MazEF-rifampicin interaction suggests a mechanism for rifampicin induced inhibition of persisters
Authors: Alexander Cyrus , Guru Ankeeta , Pradhan Pinkilata , Mallick Sunanda , Mahanandia Nimai Charan , Subudhi Bharat Bhusan , Beuria Tushar Kant
Published: October 2020

Background

Persistence is a natural phenomenon whereby a subset of a population of isogenic bacteria either grow slow or become dormant conferring them with the ability to withstand various stresses including antibiotics. In a clinical setting bacterial persistence often leads to the recalcitrance of various infections increasing the treatment time and cost. Additionally, some studies also indicate that persistence can also pave way for the emergence of resistant strains. In a laboratory setting this persistent phenotype is enriched in nutritionally deprived environments. Consequently, in a batch culture the late stationary phase is enriched with persistent bacteria. The mechanism of persister cell formation and its regulation is not well understood. Toxin-antitoxin (TA) systems have been implicated to be responsible for bacterial persistence and rifampicin is used to treat highly persistent bacterial strains. The current study tries to explore a possible interaction between rifampicin and the MazEF TA system that furthers the former’s success rate in treating persistent bacteria.

Results

In the current study we found that the population of bacteria in the death phase of a batch culture consists of metabolically inactive live cells resembling persisters, which showed higher membrane depolarization as compared to the log phase bacteria. We also observed an increase in the expression of the MazEF TA modules in this phase. Since rifampicin is used to kill the persisters, we assessed the interaction of rifampicin with MazEF complex. We showed that rifampicin moderately interacts with MazEF complex with 1:1 stoichiometry.

Conclusion

Our study suggests that the interaction of rifampicin with MazEF complex might play an important role in inhibition of persisters.