bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibitory effects of ginsenosides on S100 protein‐mediated rheumatic diseases
Authors: Cho Eun-i 
Published: April 2020

Rheumatoid arthritis(RA) is a chronic autoimmune disorder that primarily affects joints. It causes severe long‐term damage to various organ systems and results in progressive disability, systemic complications. The cause of rheumatoid arthritis is not well understood but involves a complex interplay of exogenous and endogenous inflammatory factors. Especially, numerous evidence show that S100 family also known as the pro‐inflammatory calcium‐binding protein, plays a pivotal role in the pathogenesis of rheumatic diseases. Some S100 proteins are released at the site of inflammation and participate in inflammatory signalling pathways via a variety of receptors. Therefore, these S100 proteins could be useful biomarkers for clinical joint damage in RA. Ginsenosides are a class of natural product steroid glycosides and triterpene saponins, which has a long history of use in traditional medicine. It has also been used in clinical practice for resolution of inflammation. In this study, we measured that RAW264.7 murine macrophage cell line and murine BMDMs are transfected or treated with S100 in presence of ginsenosides. The levels of cytokines were measured by ELISA. The results show that secretion of S100 proteins was inhibited by ginsenosides and pro‐inflammatory cytokines was decreased at the site of inflammation. Taken together, we suggested that pro‐inflammatory signaling of S100 proteins is inhibited by ginsenoside and this result indicated that effect of ginsenoside in S100‐mediated inflammation might be potential therapeutic target for resolving rheumatoid disease.Support or Funding InformationThis work was supported by the NRF grant funded by the Korea government (MSIP) 2019R1I1A2A01064237
Development of substitute of R‐spondin 1 for production of prostate organoid
Authors: Cha Ji Hyeon , Heo Hee Sun , Lee Seung Hwan , Lee Jong Hun 
Published: April 2020

Organoid that is organ‐like cell aggregates has both the advantages of 'in vitro' and three‐dimensional advantages of animal experiment. It is very useful for modeling disease, drug development research, and testing customized medicine.R‐spondin 1 stimulates Wnt / β‐catenin signaling to induce proliferation of organoids. Thus, high concentrations of R‐spondin 1 are essential for organoid culture. However, production of R‐spondin 1 is limited and, due to the high cost of R‐spondin 1, there are financial difficulties in the practical application of organoids. We have found an alternative to the R‐spondin 1 protein from natural compounds. We also evaluated how this natural component affects prostate organoid.We confirmed that prostate organoids were established from selected natural products. Cultures were divided into Group A with R‐spondin 1, Group B with natural products and Group C with R‐spondin 1 and natural products.In order to confirm whether organoids actually have prostate tissue characteristics, we confirmed expression of basal specific markers and, lumen specific markers including prostate specific antigen (PSA). Cyrokeratin 8 and PSA were used as luminal specific markers, and p63 and cytokeratin 5 were used as basal specific markers.This study showed that R‐spondin 1, Lgr5 antagonist, could be replaced by natural compound in organoid culture. We also could observe that natural‐derived components helped R‐spondin 1 to promote growth of prostate organoid and inhibit apoptosis. This research is expected to contribute to the research of organoids that will be developed in the future.
RNA‐Binding Protein QKI is a critical pre‐RNA splicing regulator for cardiac development and function
Authors: Liu Ying , Chen Xinyun , Xu Chen , Sun Ning , Shou Weinian 
Published: April 2020

RNA‐binding protein QKI (also known as Quaking) belongs to hnRNP K‐homology (KH)‐domain protein family and is implicated a novel regulator in pre‐RNA splicing. As previously shown, the QKI homolog how potentially regulates heart lumen formation and function in fruit fly. However, QKI’s function in mammalian heart development is largely unknown. We found that Qki specifically expressed in mouse developing hearts as early as E7.0 and this high‐level expression was maintained in the hearts to adulthood. Consistent with this finding, by using human embryonic stem cells (hESCs)‐cardiomyocyte differentiation system, QKI expression was found up‐regulated at Day 6, a critical time frame that cardiogenic progenitor transition into early cardiomyocytes. To determine the biological function of QKI in cardiogenesis, we generated hESCsQKIdef by using CRISPR/Cas9 gene editing technology and analyzed the physiological impact of QKI‐deficiency to cardiomyocyte differentiation and maturation. As expected, we did not find defect in pluripotency in hESCsQKIdef, but the contractile function was dramatically reduced in hESCsQKIdef‐derived cardiomyocytes. Based on scRNA‐seq and clustering analyses, there was no significant difference in the distribution of the clusters when compared hESCsQKIdef‐cardiomyocyte group to normal control group, suggesting that the cardiomyocyte differentiation was largely unaffected in hESCsQKIdef. However, scRNA‐seq demonstrated a handful of genes encoding sarcomere proteins were significantly altered in cardiomyocyte clusters, which included ACTN2, ACTA1, MYH7, and MYL2, etc., indicating a significant defect in cardiomyocyte maturation. Bulk RNA‐seq data not only further confirmed this notion, but also provided additional depth for detailed analysis on the QKI‐mediated pre‐RNA splicing events. In conjunction with QKI‐binding motif search and RNA immunoprecipitation (RIP) analysis, we were able to demonstrate that QKI was a key pre‐RNA splicing regulator for sets of genes involved in cardiac myofibrillogenesis and cardiac excitation‐contraction (E‐C) coupling, which ultimately impacted on cardiomyocyte maturation and function. Additional analysis of cardiac developmental defects in Qkibgeo mutant mice further validated that QKI was an indispensable splicing regulator in cardiovascular development. Our results not only revealed a novel mechanism by which QKI regulates pre‐mRNA splicing in cardiogenesis, but also implied that QKI was likely involved in the pathogenesis of certain forms of congenital heart defects and cardiomyopathies.Support or Funding InformationThis work was supported in part by National Institute of Health P01HL134599
Mid‐gestation low‐dose LPS administration results in excessive weight gain upon a Western Style Diet in female mouse offspring
Authors: Plösch Torsten , Dijkstra Dorieke J. , Verkaik-Schakel Rikst Nynke , Stojanovska Violeta , Scherjon Sicco A. 
Published: April 2020

Exposure to gestational complications can have a life‐long influence on the offspring’s health. Inflammation is observed in many complications, including preeclampsia and gestational diabetes mellitus, and is likely mediating the long‐term consequences for the offspring. Here we use a mouse model of LPS‐induced maternal inflammation to examine the consequences of gestational inflammation on offspring’s metabolic health. 25μg/kg LPS was administered to pregnant C57Bl6/J mice on gestational day 10. After weaning, all offspring was fed a semi‐synthetic control diet. At 12 weeks of age, half of the animals were switched to a Western‐Style Diet (WSD). Body composition (MRI) and glucose‐ and insulin tolerance were measured at 12 and 24 weeks of age. At 24 weeks of age, blood pressure was measured, and plasma, fat pads, liver and brain were collected and analyzed at the molecular level. Males and females were analyzed separately. LPS‐exposed female offspring showed an increase in body fat mass at 12 weeks of age, and the combination of LPS‐exposure and WSD markedly increased food intake, body weight and body fat mass in the females at 24 weeks of age. Observed changes in glucose‐and insulin tolerance, leptin and insulin levels and gene expression in liver, adipose tissue and hypothalamus were associated with body weight, not with in utero LPS exposure. In utero LPS exposure leads to increased intake of WSD in female offspring, which is likely causative of the observed increased body weight and fat mass and metabolic and molecular changes. A metabolic driver for increased food intake was not identified. Thus, in this model, gestational inflammation might lead to increased food intake through programmed behavioral traits rather that fetal programming of metabolism.Support or Funding InformationThis work was supported by ZonMw (91211053).
Calciprotein Particle Inhibition Explains Magnesium‐mediated Protection against Vascular Calcification
Authors: de Baaij Jeroen H.F. , ter Braake Anique , Eelderink Coby , Pasch Andreas , de Borst Martin , Hoenderop Joost 
Published: April 2020

Phosphate toxicity is a strong determinant of vascular calcification development in chronic kidney disease (CKD). Phosphate induces the formation of calciprotein particles (CPP), which were identified to drive the calcification process. Epidemiological data of suggests that magnesium may improve cardiovascular risk by inhibiting vascular calcification. However, the mechanism by which magnesium counteracts vascular calcification is poorly understood. Here, we investigated the effects of magnesium on phosphate and secondary crystalline calciprotein particles (CPP2) induced calcification and crystal maturation.Vascular smooth muscle cells were treated with high phosphate or CPP2 and supplemented with magnesium to study cellular calcification. The effect of magnesium on CPP maturation, morphology and composition was studied by medium absorbance, electron microscopy and energy dispersive spectroscopy. Effects of magnesium intake on aortic calcification was assessed in Klotho knock‐out mice. To translate our findings to CKD patients, the effects of magnesium on calcification propensity (T50) was measured in sera from CKD patients and healthy controls.Magnesium supplementation prevented phosphate‐induced calcification in vascular smooth muscle cells. In contrast, magnesium failed to inhibit CPP2‐induced calcification, indicating it acts before the formation of CPP2. Increased expression of osteogenic genes osteopontin and alkaline phosphatase were unchanged after magnesium supplementation. Furthermore, in a Klotho knock‐out mouse model for vascular calcification, high dietary magnesium intake prevented aortic calcification effectively. Showing the potential of magnesium in patients, magnesium dose‐dependently delayed the maturation of CPP2 by several days. In human serum, addition of 0.2 mmol/L magnesium increased T50 from healthy controls by 51 ± 5 minutes (P < 0.05) and CKD patients by 44 ± 4 minutes (P < 0.05). Each further 0.2 mmol/L addition of magnesium led to further increases in both groups.Our results demonstrate crystalline CPP2 mediate phosphate‐induced calcification. Magnesium prevents crystalline CPP2 formation and thereby prevents phosphate toxicity leading to calcification. Magnesium supplementation, even at low dosages, is a potential therapeutic strategy to reduce vascular calcification in CKD.Support or Funding InformationThis research was funded by grants from the Netherlands Organization for Scientific Research (NWO Veni 016.186.012 and VICI 016.130.668) and the Dutch Kidney Foundation (Kolff 14OKG17 and 15OP02). This work was furthermore supported by the NIGRAM2+ consortium funded by Health Holland (LSHM17034) and the Dutch Kidney Foundation (16TKI02).
Understanding the Structure of Trabecular Excrescences Found in Osteoarthritic and Non‐Osteoarthritic Joint Samples
Authors: Taylor Adam M. , Allison Kathryn , Rhodes Bryan , Kerns Jemma G. , Shepherd Rebecca F. 
Published: April 2020

The presence of novel bone structures, termed trabecular excrescences, have previously been found in surgical waste tissue samples from patients with osteoarthritis, alkaptonuria and from aged bone samples. A form of trabecular excrescence is found associated with the presence of bone marrow adipocytes and the absence of bone lining cells. We hypothesize that the adipocytes are contributing to the formation of bone like structures. This is inconsistent with the current paradigm of osteoclast‐osteoblast coupled remodelling. This study aimed to determine the topographical distribution of trabecular excrescences throughout the trabecular bone in human joint samples and to further characterise their internal structure.Joint samples were obtained as surgical waste, with informed patient consent, from 20 patients with OA (mean age = 75.3) and 20 control patients without OA (mean age = 78.5). North of Scotland NHS Research Ethics Committee (UK) approved this research. Joint samples obtained included femoral heads, humeral heads, ankles, knees and shaft of femur. Each joint sample was sliced into sagittal sections. The sections were decalcified and processed for histology. Unstained sections were viewed under fluorescence microscopy. Sections were stained with haematoxylin and eosin and with immunohistochemical staining for collagen I, collagen VI and osteocalcin.Excrescences were found in both control and osteoarthritic samples. When found, the excrescences were often clustered, with several protrusions shown along a region of trabeculae. These results demonstrate that trabecular excrescences are globally distributed throughout the femoral head, and not specific to areas of osteoarthritic change. This suggests the formation of these structures may be linked to the ageing process rather than disease progression. Immunohistochemical staining demonstrated the presence of collagen VI within the extracellular matrix of the newly formed excrescences. However, we also noted the presence of osteocalcin in ‘smoothed over’ versions of excrescences.This study has shown that trabecular excrescences are initially formed by the deposition of collagen VI by adipocytes. However, the presence of osteocalcin suggested that excrescences have the potential to be remodelled through typical osteoblast‐osteoclast remodelling. Future treatments for skeletal disorders could look to exploit the scaffold like properties of structures formed by adipocytes.Support or Funding InformationAnatomical Society PhD Studentship
Controlling the Bioluminescence Output from Shewanella woodyi using Soluble Electron Acceptors
Authors: Alsabia Sahar , Mortensen Chris , Theberge Allison , Blair Anna , Wendel Nina , Biffinger Justin 
Published: April 2020

We will present bioluminescence data from Shewanella woodyi cultures in order to correlate bioluminescence intensity with changes in the electrochemical potential in a saltwater medium using soluble electron acceptors. A relationship between the concentration of NaNO3, CoCl2, or ZnCl2 to bioluminescence intensity was confirmed using aerobic cultures of S. woodyi at 20°C with glucose as the sole carbon source. In general, increasing the concentration of nitrate, Co(II), or Zn(II) reduced the bioluminescence per cell, with complete luminescence being repressed at ≥5 mM nitrate and ≥0.5 mM Co(II). Results from cell viability fluorescent staining concluded that increasing the concentration of Co(II) or nitrate did not affect the overall viability of the cells when compared to cultures lacking Co(II) or nitrate. Our presentation will show that electrochemical potentials of < 0.2 V vs NHE repress the luminescence from the cells, but the exact mechanism is unclear. Our results indicate that the luminescence intensity from S. woodyi could be systematically reduced using these three soluble electron acceptors making S. woodyi a potential model bacterium for whole‐cell luminescence bioelectrochemical sensor applications.Support or Funding InformationUniversity of Dayton Graduate Student Summer Fellowship and Dean's Summer Fellowship programs
The Problems with the Concepts of Osmole and Osmotically Active Particles
Authors: Kuang Serena Y. , Walter Stefan , Yang Xiaoqi , Li Xiaonan 
Published: April 2020

IntroductionIn this presentation, we address and resolve the problems with the current definitions of osmole and the widely used term “osmotically active particles”. This presentation precedes and connects to our second abstract titled “The Concept of Osmolarity: Problems and Resolutions” in EB2020.MethodLogical reasoning.Results1) A simple osmosis system is first defined as S‐m‐H2O: a solution (S) compartment and a water compartment separated by a selectively permeable membrane (m) that can be either water‐permeable (only permeable to water) or SP‐permeable (permeable to water and some species of solute particles (SP)). This is a missing step to define osmole and true osmolarity. 2) The problems mentioned above can be identified in three sample definitions of osmole: “One osmole (Osm) is equal to 1 mole (mol) (6.02 × 1023) of solute particles”; “One mole of osmotically active particles is called one osmole”; and “the osmole (Osm or osmol) is a non‐SI unit of measurement that defines the number of moles of a chemical compound that contribute to a solution’s osmotic pressure”. These definitions are consistent if the m in S‐m‐H2O is water‐permeable but have different meanings when the m is SP‐permeable: the first includes osmotically ineffective, permeant SP, while the third excludes them. The second definition has a problem with the ambiguous term “osmotically active particles”: are these particles osmotically active when facing a water‐permeable membrane or facing a SP‐permeable membrane or both? Only the third definition still works when the membrane is SP‐permeable, since the moles of particles “that contribute to a solution’s osmotic pressure” are the moles of the impermeant SP regardless of whether m is water‐permeable or SP‐permeable. 3) To fix the problem with the term “osmotically active particles”, we replace it with a new term “osmotically effective SP,” defined as the physical quantity representing the amount of the impermeant (i.e., osmotic pressure (π)‐contributable) SP in the S compartment of S‐m‐H2O. 4) Osmosis does not take place in an isolated solution but in S‐m‐H2O. When the m is given, the total solute particles (TSP) of S are differentiated to be permeant SP measured using millimole (mmol) and impermeant SP measured using milliosmole (mOsm). Since the unit mOsm defined above excludes any permeant SP to a given m, the unit of osmolarity (i.e., mOsm/L) defines true osmolarity: the molar concentration of the impermeant SP in the S compartment of S‐m‐H2O.ConclusionOur definitions of osmole, osmotically effective SP, and true osmolarity are all given membrane‐dependent, system parameters of S‐m‐H2O, not owned by an isolated S. These definitions are essential but not enough to thoroughly resolve the many unaddressed problems with the current definitions of osmolarity. Our second abstract addresses these problems and offers resolutions.
Effect of Mesenchymal Stromal Cells (MSC) and Platelet‐Rich Plasma (PRP) on Rat Achilles Tendon Repair
Authors: Fagan Diana , Austin Brittany , Drew Stuart , Etty Thywill , Juzbasich Dragan , Pham Errek , Marie Hazel 
Published: April 2020

Achilles tendon rupture requires long recovery times and has the possibility of re‐rupture. The goal of this project is to improve healing in surgically repaired Achilles tendons in a Lewis rat model. Full thickness surgical transections were made 6 mm proximal to the calcaneus bone. Following suture repair, a 20 x 30 mm piece of CollatapeTM was wrapped around the tendon in groups receiving collagen and the skin closed. MSC or PRP or both (100 ml) was injected subcutaneously adjacent to the incision. Both test Achilles and control Achilles from the same animal were harvested and the biomechanical properties determined using a standard force‐extension analysis of tensile properties after 1 week or 2 weeks. A significant improvement in Strain at UTS and Strain at Failure (calculated as percent of control tendon) was seen at 1 week in rats receiving CollatapeTM, plus PRP and MSC. This project was approved by the Northeast Ohio Medical School IACUC.Support or Funding InformationSupported by Mercy Health, Youngstown and Youngstown State University.
Chronic Intermittent Hypoxia Alters the Chloride Gradient in Median Preoptic Nucleus (MnPO) Neurons of Rats
Authors: Farmer George E , Little Joel T , Bachelor Martha E. , Rybalchenko Nataliya , Yuan Joseph , Cunningham J. Thomas 
Published: April 2020

Chronic intermittent hypoxia (CIH) is an animal model that simulates the hypoxemia associated with obstructive sleep apnea (OSA). Rats exposed to CIH exhibit increased sympathetic tone and reactivity and a persistent elevation in blood pressure during periods of normoxia, similar to patients with OSA. In MnPO neurons, angiotensin II type 1 receptor knock down (AT1aR KD) mediated reductions in GABAa inhibition are exacerbated to the point where GABAa activation is excitatory following CIH. Here, we use the genetically encoded ratiometric Cl‐sensor, ClopHensorN, to monitor the chloride flux of PVN projecting MnPO neurons in normoxic (Norm) and CIH treated rats in response to GABAa receptor activation.Using isoflurane (2–3%) anesthesia, male Sprague‐Dawley rats (250–350g) received microinfusions (0.4 μL) of a retrograde AAV9‐Cre in the PVN and a DIO‐ClopHensorN in the MnPO. After recovery, rats were subjected to 7 days of CIH (0800–1600 hrs) or Normoxia. The CIH protocol consisted of 6 min cycles (3 min 21% O2, 3 min 10% O2) repeated 10x/hr for 8 hours (during the normal inactive/sleep phase) on 7 consecutive days. For ClopHensorN measurements, rats were anesthetized with isoflurane (2–3%) and coronal slices (300 μm) containing the MnPO were cut using standard in vitro slice procedures. Image sets were captured every 3 sec. Cl‐flux was determined from the ratio of GFP (ex 488 nm, em 500–550 nm) to tdTomato (ex 594 nm, em 650–700 nm) fluorescence in response to 10 s focal application of muscimol (100 uM).Twelve rats (6 Norm, 6 CIH) were used for ClopHensorN studies. In MnPO neurons from CIH treated rats (n = 632), 20.1% showed a decrease in the ratio of GFP to tdTomato fluorescence (n = 127) while 0.3% showed an increase in this ratio (n = 2) indicative of Cl‐efflux. In MnPO neurons from Norm rats (n = 482), 41.9% showed a muscimol dependent decrease in the ratiometric Cl‐signal (n = 202) with 0 cells showing an increase. Furthermore, the magnitude if muscimol dependent decreases in the Cl‐signal were reduced in the CIH treated rats suggestive of reduced GABAa mediated inhibition. In a separate groups of rats, Western blot analysis showed that CIH increased the expression of pKCC2 (n = 6) in punches containing the MnPO compared to MnPO punches from Norm controls (n = 5) injected with Scr. CIH/AT1a KD rats showed decreased pKCC2 (n = 6) compared to CIH/Scr but increased pKCC2 in Norm/AT1a KD rats (n = 6) compared to Norm/Scr rats.The results demonstrate that CIH alters the Cl‐flux of PVN projecting MnPO neurons and these changes are likely due to altered function of KCC2. These changes may contribute to the increased sympathetic tone and reactivity, as well as the persistent hypertension associated with CIH.Support or Funding InformationSupported by P01 HL088052