bluuebunny/crossref_metadata_2025_split huggingface dataset:
Luminal Succinate and Acidosis: Effects on Blood Pressure
Authors: Hering-Smith Kathleen , Huang Weitao , Teran Fred , Sato Ryosuke , Hamm L. Lee 
Published: April 2020

Succinate (Suc) is well known as a Krebs cycle intermediate but extracellular Suc has been associated with a variety of pathologic processes. A distinct GPCR receptor for Suc (Sucnr1) was identified in the kidney and some (but not all) studies have documented that Suc infusions increase blood pressure (BP) via paracrine signaling. Luminal Suc has been reported to activate Sucnr1, stimulate renin release and alter BP in certain circumstances.In the nephron, apical NaDC1 (Na+ dicarboxylate cotransporter) is located only in the proximal tubule, reabsorbs filtered citrate and Suc, and is upregulated in acidosis. Thus NaDC1’s regulation is important in preventing calcium stones and promoting acid‐base homeostasis via effects of citrate reabsorption but its role in BP regulation is not known. Our purpose was to explore the role of NaDC1, luminal Suc, and acidosis in BP regulation.To address these issues, we used NaDC1 KO and WT mice on normal diet or 72 hr acid diet (0.4 M HCl). In initial clearance (CL) studies, KO mice had increased urinary Suc and citrate but no statistical difference in BP even with the infusion of 1 mmol/kg Suc. Suc infused WT did not have increased BP in CL. Acid diet reduced urinary Suc in WT, but had no consistent effect on BP in either KO or WT in CL.Since recent reports suggested that Suc may affect BP exclusively during more active periods (nocturnal in mice), we additionally examined BP using telemetry (BP/T) for 24/7 monitoring. During 8 PM‐5 AM on a normal diet mean arterial pressure (MAP) was significantly higher in KO than in WT. Acidosis appeared to lessen these BP/T differences. As an example of this, Figure
1 shows MAP at selected time points in a 24 hour cycle, 8 AM, 9 PM, 5AM and 8AM.To determine whether Sucnr1 gene expression alterations could influence these results, droplet digital PCR (Bio Rad) was performed. Data are expressed as copy numbers of target gene in 1 ng PCR reaction. Thus with ddPCR we determined on normal diet, Sucnr1 expression levels in KO (1343.6 ± 51) are significantly increased from that in WT (1052.8 ± 50, p< 0.007). Furthermore, on 72 hr acid diet the expression of Sucnr1 in KO (1039.6 ± 33) fell while in WT increased to 1148 ± 50, p <0.006).In sum, on normal diet, Sucnr1 expression is significantly greater in NaDC1 KO vs WT and in chronic acidosis expression decreases in KO but increases in WT. On normal diet MAP in KO is significantly higher than in WT while acidosis diminishes these differences (by slightly lowering KO MAP but raising slightly WT MAP).These diverse findings illustrate that NaDC1 (and presumably luminal Suc) can regulate BP but that this regulation varies with time of day (or associated factors such as activity) and acidosis.Support or Funding InformationNIH, DCI PTRF, LA CaTSMAP differences in NaDC1 KO mice vs WT mice on a normal dietFigure 1
Clathrin‐mediated phagosome resolution regenerates lysosomes and maintains degradative capacity of macrophages after phagocytosis
Authors: Botelho Roberto , Fountain Aaron , Lancaster Charlene , Terebiznik Mauricio 
Published: April 2020

Phagocytosis is the selective internalization of large particles and is critical for immunity, development and tissue homeostasis. The conversion of phagosomes into microbicidal compartments is a highly regulated process, which has been studied in great detail. Conversely, the fate of phagolysosomes, after their cargo is digested, is poorly understood. By utilizing fluorescent bacteria and labelling phagosomal markers, we followed mature phagolysosomes to their resolution. Despite previous assumptions, phagolysosomes are not exocytosed, but instead undergo a continuous dismembering process resulting in the shedding of myriads of vesicles containing degraded cargo. These phagosome‐derived vesicles resemble lysosomes, based on the acquisition of lysosomal markers, acidification and fusogenicity with newly formed phagosomes. Combining pharmacological and genetic strategies, we found that phagosomal fragmentation is dependent on cargo degradation, clathrin and the integrity of the actin and microtubule cytoskeleton. In addition, we provide evidence that phagosome‐derived vesicles are required to recycle lysosomal components allowing for the formation of new phagosomes and consequently maintaining the capacity of macrophages to ingest targets. Thus, we propose that phagolysosome resolution recycles cellular resources for the continuous generation of new phagosomes and the prevention of this phase will critically impair the immune and housekeeping functions of the phagocytes.Support or Funding InformationCanadian Institutes of Health Research Canada Research Chairs Program Ryerson University Natural Sciences and Engineering Council of CanadaClathrin mediates phagosome resolution. A. Macrophages engulfed mRFP1‐expressing E. coli. At 1 h post‐phagocytosis, phagosomes are rod‐shaped, like E. coli and mRFP1 is mostly associated with phagosomes. After 4 h of phagosome maturation, phagosomes shrunk and mRFP1 is found in small vesicular compartments. Clathrin inhibitors, pitstop and ikarugamycin, impair phagosome fragmentation. B. Quantification of phagosome‐derived vesicles, where b is significantly different from other conditions.Figure 1
Development of Potent Inhibitors of the Human Microsomal Epoxide Hydrolase
Authors: Singh Nalin , Barnych Bogdan , Negrel Sophie , Zhang Yue , Magis Damien , Roux Capucine , Hua Xiude , Ding Zhewen , Morisseau Christophe , Tantillo Dean J. , Siegel Justin , Hammock Bruce D. 
Published: April 2020

Microsomal epoxide hydrolase (mEH) hydrolyzes a wide range of epoxide containing molecules. Although involved in the metabolism of xenobiotics, recent studies associate mEH with the onset and development of some diseases. This phenomenon is partially attributed to the significant role mEH plays in hydrolyzing endogenous lipid mediators, suggesting more complex and extensive physiological functions. In order to obtain pharmacological tools to further study the biology and therapeutic potential of this enzyme target, we describe the development of highly potent inhibitors of the human mEH with IC50 values in the low nanomolar range, around 2 orders of magnitude more potent than previously obtained mEH inhibitors. Rationalization of binding through docking calculations of the inhibitors to a mEH homology model revealed that an acetamide, with a bulky aliphatic substituent at the α‐position, as well as an aryl ring 3–4 bonds away function as key pharmacophore units. No interaction was observed with the thioether moiety, suggesting it can be replaced by more metabolically stable (e.g. methylene) groups in future studies.Support or Funding InformationNational Institute of Health (NIH) R35 ES030443 and R01 GM076324‐11 grants; National Institute of Environmental Health Sciences (NIEHS) Superfund Research Program P42 ES004699 grant; National Science Foundation Awards 1827246, 1805510 and 1627539; Computation support from Rosetta Commons and the US National Science Foundation’s XSEDE programCompound 62: The most potent mEH inhibitorFigure 1Modeling of key molecular interactions in one binding mode. Left: interactions between amide group of 62 with D226, Y299 and Y374 side chain. Right: π–π stacking between 62 and W227.Figure 2
Mapping the Multi‐Modal Distribution of Craniofacial Phenotypes in NOSIP Mutants
Authors: Richbourg Heather A. , Saliou Alexa , Devine Jay , Green Rebecca , Hoffmeister Meike , Oess Stefanie , Hallgrimsson Benedikt , Marcucio Ralph 
Published: April 2020

Nitric Oxide Synthase Interacting Protein (NOSIP) is a E3 mono‐ubiquitin ligase that regulates the activity of protein phosphatase 2A. Deletion of NOSIP leads to severe and variable malformations of the craniofacial complex. Our goal was to use NOSIP mutant mice to understand of mechanisms of phenotypic variation. The phenotype of an individual embryo is the product of the developmental trajectory that it previously traversed. We propose that this concept of canalization is essential for understanding the etiology of complex phenotypic outcomes. We have previously shown that the brain patterns the face, so sequential perturbations affecting the brain and face may create unique developmental trajectories that each embryo transits. We tested this by creating a 3D morphospace of NOSIP heterozygous and homozygous mutants to determine if embryos occupy unique morphospace regions based on affected craniofacial structures. Embryos between embryonic day 10.5 (E10.5) to E17.5 were collected, fixed, and imaged (mCT). Anatomical features were reconstructed using Avizo. NOISP expression appears to be restricted to the neuroepithelium, lower jaw mesenchyme, and mesenchyme and epithelium in tissue adjacent to the frontonasal ectodermal zone between stage E10.5‐11.5. Embryos with a single telencephalic lobe had more severe facial malformations, including micrognathia and obstructed airway without significant changes in early cell apoptosis (via TUNEL staining). Embryos with normal telencephalic vesicle separation had less severe facial phenotypes, including left‐right mild asymmetry and clefting. Overall, it appears that NOSIP‐KO creates a multi‐modal distribution of craniofacial phenotypes along the developmental timeline by affecting early brain development, suggesting that understanding canalization may provide insight into the complexity of genotype‐phenotype map (Fig.
1).Support or Funding InformationFunding was provided by NIH F32DE08476 and R01DE019638 grants.Schematic of the changing phenotypic landscape describing the multi‐modal distribution of craniofacial phenotypes within NOSIP+/− and NOSIP−/− embryos compared to wildtype (light blue).Figure 1
Novel PAR2 antagonist ameliorates progression of lupus nephritis in NZB/Z F1 mice
Authors: Seo Yohan , Mun Chin Hee , Park Yong-Beom , Lee Sang-Won , Namkung Wan 
Published: April 2020

Lupus nephritis (LN) is, the most frequent phenotype in patients with systemic lupus erythematosus (SLE), has a high rate of progression to end‐stage renal disease, even with intensive induction and maintenance therapies, approximately 20% of patients with LN progress to end‐stage renal disease. Recent evidence suggests that protease‐activated receptor‐2 (PAR2) is a therapeutic target for glomerulonephritis. In this study, we identified a novel PAR2 antagonist and evaluated the effect of the PAR2 antagonist on LN in lupus‐prone mouse model. To identify novel PAR2 antagonist, a cell‐based high‐throughput screening was performed. The effect of PAR2 antagonist on podocytes was observed in human podocyte cell line and LN was evaluated in NZB/W F1 mice. We identified punicalagin (PCG, a major polyphenol in pomegranate) as a novel potent PAR2 antagonist. In human podocyte cell line, PCG potently inhibited PAR2 (IC50 = 1.5 ± 0.03 μM) with ~30‐fold selectivity for PAR2 over PAR1 and significantly reduced the PAR2‐mediated activation of ERK1/2 and NF‐κB signaling pathway. In addition, PCG strongly decreased PAR2‐induced increases in ICAM‐1, VCAM‐1, IL‐6, IL‐8, IFN‐γ, and TNF‐α expression. Notably, in NZB/W F1 mice, which spontaneously develop severe autoimmune syndrome resembling human SLE and LN, the intraperitoneal administration of PCG significantly alleviated kidney injury and splenomegaly and reduced proteinuria and renal ICAM‐1 and VCAM‐1 expression. Our results suggest that PCG has beneficial effects on LN via inhibition of PAR2, and PCG is a potential therapeutic agent for LN.Support or Funding InformationThis research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2017R1D1A1B03029050), a faculty research grant of Yonsei University College of Medicine (6‐2016‐0159) to Sang‐Won Lee, NRF (2015R1D1A1A01057695 and 2018R1A6A1A03023718) to Wan Namkung, and NRF (2019R1I1A1A01061117) to Yohan Seo. This research was supported (in part) by the Yonsei University Research Fund (Post Doc. Researcher Supporting Program) of 2019 (project no.: 2019‐12‐0013).Schematic overview of inhibitory effect of Punicalagin (PCG) on lupus nephritis in human podocytes and NZB/W F1 mice.The inhibition of PAR2 by PCG reduced PAR2‐induced ERK1/2 and NF‐κB activation in human podocyte cell line. Notably, PCG ameliorated kidney injury, proteinuria, and splenomegaly in NZB/W F1 mice and reduced ICAM‐1 and VCAM‐1 levels via PAR2 inhibition.Figure 1
Pharmacological inhibition of soluble epoxide hydrolase alleviates chronic neuropathic pain induced anhedonia symptoms: differential role of AhR and TSPO signaling
Authors: Shan Li , Ailin Luo , Dongyu Hua , Gaofeng Zhan , Hammock Bruce D. , Hashimoto Kenji , Chun Yang 
Published: April 2020

Patients who suffer from chronic pain have a higher incidence to develop depression, which needs effective treatments. TPPU, a soluble epoxide hydrolase (sEH) inhibitor, was reported to have analgesia and antidepressant effects in animal models, suggesting its potential improvement in this comorbidity. Here, we sought to investigate: (1) the occurrence of anhedonia symptoms in mice with chronic neuropathic pain and the sEH protein expression in different tissues, including central nervous system and peripheral tissues; (2) the effects of TPPU on nociceptive and anhedonia behaviors in anhedonia susceptible mice, and (3) the involvement of Aryl hydrocarbon receptor (AhR) signaling and translocator protein (TSPO) signaling as possible targets for this comorbidity. Spared nerve injury (SNI) was constructed to mimic chronic neuropathic pain (CNP) model, and that mice in SNI group were classified into chronic pain with or without anhedonia phenotype via hierarchical cluster analysis of sucrose preference test (SPT) results. Protein levels of sEH were increased in medial prefrontal cortex (mPFC), hippocampus, spinal cord, liver, kidney and gut, but not in anterior cingulate cortex (ACC), nucleus accumbens (NAc), striatum, cerebellum, heart, muscle and vessel in anhedonia susceptible mice as compared with sham or anhedonia resilient mice. Chronic treatment with TPPU (3 mg/kg, orally, once per day for 7 days) improved withdraw threshold and SPT scores in anhedonia susceptible mice. Interestingly, the antidepressant, but not analgesia effect, of TPPU seems to be pharmacologically abrogated by pretreatment with AhR agonist (FICZ, orally, 100 μg/kg) via regulating inflammatory cytokines and CYP1A1 levels in the serum as well as neuroinflammation in the microglia and astrocytes. Interestingly, the classical TSPO pathway, a crucial factor in the analgesia and antidepressant effects of TPPU, failed to show any difference in subjects who were treated with drugs targeted at the AhR. Collectively, our findings indicate that TPPU may alleviate depression‐like symptoms in CNP mice through two independent pathways: (1) the direct antidepressant effects via regulating neuroinflammation mediated by AhR, and (2) the improvement of allodynia and depressive symptoms via the TSPO signaling and downstream neurosteroids. Thus, AhR provides a potential therapeutic target for treatment of depression induced by chronic pain, and strategies targeting TSPO signaling could be a vital approach to alleviate the comorbidity of chronic pain and depression.Support or Funding InformationThis work was supported , in part, by grants from the National Natural Science Foundation of China (to A.L., 81771159 and 81571047; to C.Y., 81703482)The overview of our manuscript.Figure 1
Structural Characterization of the Apoptosis Signal‐regulating Kinase 1 (ASK1) Domain Responsible for Thioredoxin Binding and its Complex
Authors: Obsil Tomas , Psenakova Katarina , Hexnerova Rozalie , Srb Pavel , Obsilova Veronika , Veverka Vaclav 
Published: April 2020

Apoptosis signal‐regulating kinase 1 (ASK1) is a mitogen‐activated protein kinase kinase kinase 5 implicated in p38 mitogen‐activated protein kinase and c‐Jun N‐terminal kinase signaling cascades. ASK1 responds to a wide range of signaling cues and is dysregulated in cancer, neurodegeneration and cardiovascular diseases. The catalytic activity of ASK1 is tightly controlled through oligomerization and binding of several cofactors, among which thioredoxin stands out as the most important ASK1 inhibitor. Interestingly, only the reduced form of thioredoxin inhibits ASK1 by direct binding to its N‐terminal domain and oxidation‐driven thioredoxin dissociation is the key event in oxidative stress‐mediated ASK1 activation. Despite extensive efforts, the molecular basis of this key cellular regulatory mechanism has remained unknown until now, primarily due to the dynamic properties of the ASK1 N‐terminal domain, which has prevented its structural characterization by X‐ray crystallography. Here, we report the structural characterization of the ASK1 domain responsible for thioredoxin binding and of the ASK1: thioredoxin complex using NMR spectroscopy and chemical cross‐linking, which provides the molecular basis for complex dissociation under oxidative stress conditions. Our data reveal that the N‐terminal domain of ASK1 adopts a fold resembling the thioredoxin structure while retaining substantial conformational plasticity at the thioredoxin‐binding interface. Although oxidative stress induces relatively moderate structural changes in thioredoxin, we show that these changes lead to a substantial conformational rearrangement of the thioredoxin‐binding interface on ASK1 driven by the formation of intramolecular disulfide bridges. Lastly, we demonstrate that the cysteine residue at the position 250 of ASK1 is the key element of this molecular switch.Support or Funding InformationThis work was supported by the Czech Science Foundation (Project 19‐00121S), the Grant Agency of the Charles University (Project 368216), the Czech Academy of Sciences (Research Projects RVO: 67985823 of the Institute of Physiology) and partially also by the European Regional Development Fund; OP RDE; Project: ChemBioDrug (No. CZ.02.1.01/0.0/0.0/16_019/0000729).Structural model of the ASK1‐TBD:TRX complex calculated using the program HADDOCK with the NMR‐based structural model of ASK1‐TBD, and crystal structure of reduced TRX1 (PDB ID: 1ERT).Figure 1
Transient receptor potential vanilloid 4 (TRPV4) channels are essential for alveolar epithelial cell function
Authors: Dietrich Alexander , Chao Yu-Kai , Kannler Martina , Krasteva-Christ Gabriela , Rajan Suhasini , Yildirim Ali Oender , Brosien Monika , Schredelseker Johann , Weissmann Norbert , Grimm Christian , Gudermann Thomas W. , Weber Jonas 
Published: April 2020

Ischemia‐reperfusion‐induced edema formation as one of the most common and significant causes of morbidity and mortality after lung transplantation can be mimicked ex‐vivo in isolated perfused mouse lungs (IPL). Transient receptor potential vanilloid 4 (TRPV4) is a non‐selective cation channel expressed in lung endothelium where increased channel activity has been shown to compromise endothelial barrier function. Here we show enhanced edema formation in TRPV4‐deficient (TRPV4−/−) IPL compared to wild‐type (WT) controls in response to ischemia and reperfusion, indicating a protective role of TRPV4 to maintain the alveolar epithelial barrier. By immunohistochemistry or mRNA profiling, we detected TRPV4 in bronchial epithelium, alveolar type I (ATI) and alveolar type II (ATII) cells. Genetic ablation of TRPV4 resulted in reduced expression and plasma membrane insertion of the water conducting aquaporin‐5 (AQP‐5) channel in ATI cells compared to WT mice. Analysis of isolated primary TRPV4‐deficient ATII cells revealed a reduced expression of surfactant protein C (SP‐C), which is essential for decreasing surface tension and alveolar fluid homeostasis. The TRPV4 activator GSK1016790A induced increases in current densities only in WT but not in TRPV4−/− ATII cells. Although the ability of TRPV4−/− ATII cells to differentiate to ATI cells was unchanged, epithelial mesenchymal transition (EMT) of TRPV4−/− ATII cells was impaired and migration and cell barrier function of TRPV4−/− ATI cells was reduced. Moreover, TRPV4−/− lungs of adult mice developed significantly larger mean chord lengths and altered lung function compared to WT lungs. Therefore, our data highlight novel essential functions of TRPV4 channels in alveolar epithelial cells and in the protection from edema formation.Support or Funding InformationThis study was funded by the Deutsche Forschungsgemeinschaft (TRR 152 (AD, GKC, CG), GRK 2338 (AD, CG)) and the German Center for Lung Research (DZL) (AD, MB, NW, TG).Phenotypes in alveolar epithelial cells after ablation of TRPV4. See text for details.Figure 1
Learning Anatomy Through Yoga
Authors: Sullivan Alyssa , Vasudevan Anita , Fox Glenn M. , Prall Cristina R. , Alsup B. Kathleen 
Published: April 2020

The aim of this project is to provide a unique tool for anatomy students of all levels to apply common physical activities in the process of learning anatomical structures and functions. The practice of yoga was chosen to demonstrate the dynamic anatomical components of the upper and lower limbs through a variety of poses. This is explored through a series of videos: ‘Learning Anatomy through Yoga’. This series was developed with a fourth year medical student at the University of Michigan, who is also a certified yoga instructor. During the musculoskeletal sequence in April, first year medical students will assess the efficacy of this resource via surveys. In the future, the ‘Learning Anatomy through Yoga’ video series will be part of a broader project that demonstrates functional anatomy in everyday physical activity, such as weight lifting, rowing, pilates, etc.Instructor demonstrating specific attachment points on skeletonFigure 1