bluuebunny/crossref_metadata_2025_split huggingface dataset:
Epitope Characterization and Design of Epitope Binding Proteins for Idiopathic Membranous Nephropathy: New Tools for Autoimmune Kidney Disease Therapy
Authors: Goldman Liam , Stocks Omar , Stoddard Shana V. 
Published: April 2020

In autoimmune diseases antibodies incorrectly attack normal healthy cells, which can promote degradation of tissues and can lead to long‐term disease and death. Modern clinical therapies treating autoimmune disease include the use of immunosuppressive medicines. Although this therapy has shown success in slowing down the progression of autoimmune disorders, this treatment diminishes the patient's entire immune system's ability to fight off normal pathogens. This highlights the need for more specific treatment options for autoimmune disease that do not weaken the patient's entire immune system. Primary membranous nephropathy (PMN) is a kidney specific autoimmune disease currently being treated with immunosuppressive medicines. In patients with PMN, seventy percent of individuals produce autoantibodies that target the phospholipase A2 receptor (PLA2R). The goal of this research is to design PLA2R specific binding proteins (epitope caps) that will prevent the PLA2R autoantibodies from binding to the C‐type lectin domain number seven (CTLD7) on this antigen. Using Epitopia and EPCES, three possible epitope sites were identified and characterized. Binding strength of the binding proteins to the CTLD7 was measured through Rosetta Online Server that Includes Everyone. In silico mutations were made to the Monobody template 5FXB. Currently, we have produced three epitope caps (5FXB‐001‐LVG, 5FXB‐002‐LVG, 5FXB‐003‐LVG) with binding scores that are below out targeted binding score of −5.0 Rosetta energy units (REU). Currently protein expression and purification is underway to permit further evaluation of the epitope caps using ELISA assays to determine experimental binding proficiency. Furthermore, this research could provide patients with more specific methods of treatment for autoimmune disease than the contemporary treatment options such as, immunosuppressive therapies.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mechanism of a membrane ‘transceptor’: A transporter and a receptor in one
Authors: Gupta Meghna , Stroud Robert 
Published: April 2020

Certain transporter‐like membrane proteins also act as receptors for the availability of external nutrient content in eukaryotic cells, hence called ‘transceptors’. The concept came into the picture when several transporter‐like receptors were discovered in eukaryotes. They can be classified as transporting (example yeast Mep2, Gap1, Pho84, mammalian SNAT2) and non‐transporting (example yeast Snf3, Rgt2, mammalian SGLT3) transceptors based on their ability to transport the substrate. I sought to determine the mechanisms by which a key phosphate transceptor combines these dual functions at the level of atomic structure and biochemistry. Phosphate is an essential macronutrient for structural and metabolic requirements of all eukaryotic cells; for nucleotide and lipid biosynthesis, cell signaling by phosphorylation and energy storage (ATP), and because of its impact, pathways inside the cell must respond to availability even outside the cell, in a regulated fashion. I selected the phosphate transceptor PiPT from Piriformospora indica, for which structure is available in atomic details, the first for any transceptor. This enabled the mutagenesis of regions as a test of their transport and/or receptor function, and further structure analysis aimed at probing the signaling pathway across the membrane. Compounds known to inhibit transport and/or activate the receptor function were tested and used to distinguish between the two functions. Site‐directed mutagenesis based on the available structure was exploited to determine the role of residues that bind phosphate in the central transport site and to define the alternate access to both sides of the membrane, and the separate pathway for protons and the way their transport is absolutely coupled to phosphate transport. Transport assays and biochemical assays probing the intracellular signaling response for the mutants studied assisted in determination of a possible mechanism of transport and receptor function.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of interleukin‐1 receptor‐associated kinase 4 in drug addiction
Authors: Wu Ruyan , Liu Jian‐Feng , Vu Jimmy , Johnson Bernard , Li Jun‐Xu 
Published: April 2020

Drug addiction remains an unmanageable and costly disease. Apart from neuronal adaptation, growing recognition arises that glial proinflammatory activation importantly contributes to the rewarding effects of multiple drugs of abuse. Recent studies suggest the central importance of Toll‐like receptor 4 (TLR4), a candidate neuroimmune therapeutic target for drug addiction. While Interleukin‐1 receptor associated kinase 4 (IRAK4) plays a crucial role in TLR4 mediated innate immunity, there is no further studies support its functioning in drug addiction. We hypothesized that drug‐taking activates IRAK4 and induces reinforcing effects, and disruption of IRAK4 signaling attenuates the addictive behaviors. In the present study, IRAK4 and IRAK1 phosphorylation after morphine and cocaine self‐administration was evaluated using western blotting. The role of IRAK4 in morphine self‐administration and cue‐induced reinstatement was examined with its inhibitor, PF06650833. Moreover, local pharmacological manipulation was conducted to determine the role of IRAK4 in the nucleus accumbens (NAc) core in the cue‐induced reinstatement of morphine seeking. We found that morphine self‐administration significantly increased the phosphorylation of IRAK4, but not IRAK1, both in NAc and VTA. However, neither cocaine short access nor long access self‐administration had any effect on the phosphorylation of IRAK4. Systemic administration of PF06650833 significantly attenuated cue‐induced reinstatement of morphine seeking without affecting the spontaneous locomotion in rats, and microinjection of PF06650833 into NAc core sufficiently decreased cue‐induced morphine reinstatement. These results demonstrated that modulation of IRAK4 activity regulates the cue‐induced morphine reinstatement, and suggested it as a novel candidate target in treating drug addiction.Support or Funding InformationThis work was supported by NIDA [Grant R21DA040777].This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The role of autophagy in mesenteric arteries from hypertensive rats
Authors: Kwon Youngin , Choi Soo‐Kyoung , Byeon Seonhee , Lee Young‐Ho 
Published: April 2020

Autophagy is an intracellular degradation system that disassembles cytoplasmic components through autophagosomes with lysosomes. Autophagic dysregulation may be related vascular diseases including hypertension, pulmonary hypertension and atherosclerosis. Recently, there are increasing interests about involvement of autophagy in survival and function of vascular smooth muscle cells. In hypertensive rat, impaired relaxation and augmented contraction elevate peripheral resistance. We observed expression levels of autophagic markers beclin1, LC3I/II and p62 in mesenteric arteries and aorta from control rats and hypertensive rats. Significantly increased beclin1 and LC3II were observed in hypertensive rats compared with control rats. We checked the expression level of p62, autophagosome cargo protein, which is degraded by autolysosomes. The level of p62 was significantly decreased in hypertensive rats. In this study, we assumed that alterations in autophagic flux change function of mesenteric arteries from hypertensive rats.Support or Funding InformationThis work was supported by the Basic Science Research Program of the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF‐2018R1D1A1B07041820) and Brain Korea 21 PLUS Project for Medical Science, Yonsei University.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Functional significance of O‐GlcNAcylation of ICln in the regulation of cellular volume
Authors: Remigante Alessia , Costa Roberta , Civello Davide , Morabito Rossana , Nagy Tamas , Marino Angela , Dossena Silvia 
Published: April 2020

O‐GlcNacylation (O‐GlcNAc) is a post‐translational modification of serine or threonine residues of nuclear and cytoplasmic proteins and occurs via conjugation to a single monosaccharide, the N‐acetylglucosamine. O‐GlcNAc modification intervenes in a wide variety of physiological and pathological processes, and negatively affects the regulation of cellular volume with a molecular mechanism still unknown. Recently, the protein ICln, crucial in the activation of a chloride conductance (IClswell) after anisosmotic cell swelling, has been found to be O‐GlcNacylated. Mass spectrometry and bioinformatics show multiple O‐GlcNAc modification sites, of which the impact on ICln function is unknown. To explore the functional significance of O‐GlcNAc modification of ICln, the wild type and different mutant forms of ICln have been expressed in a heterologous system and characterized by whole‐cell patch‐clamp in the presence of normal or elevated O‐GlcNAc levels. The results show that: I) O‐GlcNAc elevation suppresses the ICln‐induced current; II) IClnT223A is functional and sensitive to O‐GlcNAc elevation; III) IClnS193X loses most of its activity, though the residual current is sensitive to O‐GlcNAc elevation; IV) IClnS67A is functional but insensitive to O‐GlcNAc elevation; V) the IClnS67T‐induced current is lower compared to the wild type, and is no longer responsive to O‐GlcNAc elevation. Overall, the evidence obtained indicates that O‐GlcNAcylation of ICln at the level of Serine 67 leads to suppression of the ICln‐induced current and may disclose the mechanism by which O‐GlcNAc elevation alters the regulation of cellular volume. We suggest that the protein ICln may represent a novel target in the prevention or treatment of pathological states characterized by chronically elevated O‐GlcNAcylation levelsThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mast Cells and Macrophages of Rats lungs in an Acute Model of Fat Embolism
Authors: Lin Jane Hua‐fang , Egdebe Joy , Hamidpour Soheila , Omoscharka Evanthia , Haferkamp Thomas , Lind Taylor , Spaedy Abigail , Silswal Neerupma , Pour Mohammad , Poisner Alan , Moneghan Paula , Molteni Agostino 
Published: April 2020

IntroductionFat embolism (FE) by IV injection of Triolein (T) causes vasculitis, septal inflammation and fibrosis in rat lungs at 48 hours, which is mitigated by two drugs acting on the renin‐angiotensin system (RAS) Captopril and Losartan. An increase in mast cells is noted following administration of T, with an even higher increase by concomitant treatment with Captopril and significant induction by Losartan. This study examines the presence and number of macrophages in rat lungs after T treatment with or without addition of Captopril and Losartan.MethodsSprague‐dawley rats (280–300g) were treated with 0.2 ml IV T or saline. One hour later 0.2 ml of Saline, Captopril, or losartan was administered IP in the 3 T subgroups. All rats were necropsied 48 hours later under isoflurane anesthesia. Lungs were fixed in 10% formalin and stained with: 1. hematoxylin eosin for morphology, 2. trichrome for fibrosis, 3. CD 117 for mast cells and 4. CD 68 for macrophages. Two pathologists unaware of slide identity reviewed ten random photographs of all slides at 400x for macrophage count and histopathological evaluation. Slide photographs and mast cell count was performed by 2 other pathologists.ResultsAs previously reported, T injection induced pulmonary vasculitis, septal inflammation and fibrosis which was markedly reduced by the RAS inhibitors Captopril and Losartan. Contrary to the statistically significant increase of mast cells after these 3 treatments, macrophage counts were only increased but minimally by T and T + Captopril. T + Losartan administration yielded a macrophages count similar to that of the saline controls.Two types of macrophages were observed: one composed of small cells with compact cytoplasm and nucleus, similar in their size to the size of the mast cells and a second type with a larger vacuolated cytoplasm. The number of cells of this second group was rather small, so that any increase of these cells could not influence the macrophages total number.ConclusionBoth mast cells and macrophages play a determinant role in the pathogenesis of the inflammatory and fibrotic pulmonary damage after T fat embolism. A different response to the injury was observed between the numbers of these two series of cells in the earlier phase of the injuries. The difference also observed in mast cells versus macrophages number present following T + Captopril and T + Losartan administration, suggesting a diversely regulated pharmacologic activity of the Renin Angiotensin System of the two drugs in the protection of the fat embolism mediated damage.Support or Funding InformationFunded and supported by the Catherine T. Geldmacher Foundation St. Louis, Missouri.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of phosphodiesterase‐4D reverses memory deficits and depression‐like effects via cAMP signaling in mouse models of Alzheimer's disease
Authors: Zhang Hanting , Wang Hao , Zhang Fangfang , Li Bing , Zhou Yanmeng , Yu Haiyang , Xu Ying , O'Donnell James 
Published: April 2020

Depression is highly correlated with Alzheimer's disease (AD), but treatment for the comorbidity of both is lacking. Phosphodiesterase‐4 (PDE4), an enzyme that catalyzes the hydrolysis of cyclic AMP (cAMP), has been considered as a promising target for treatment of both memory loss in AD and depression. Using mouse models of AD and potent PDE4 inhibitors and PDE4‐subtype knockout mice, we demonstrated that PDE4D plays a role in the comorbidity of memory loss and depression in AD. Treatment with the PDE4 inhibitor rolipram or roflumilast reversed memory deficits in novel object recognition and Morris water‐maze tests in APP/PS1 double transgenic and 3xTg‐AD mice, which are widely used models for AD. Similarly, in the tail‐suspension and forced‐swimming tests, the PDE4 inhibitors reversed the decreased immobility in AD mice, suggesting antidepressant‐like effects. The effects of PDE4 inhibitors were mimicked in the tests using mice deficient in PDE4D, which displayed memory‐enhancing and antidepressant‐like effects, relative to the WT mice microinfused with beta‐amyloid peptide 1–42 into the hippocampus. In addition, the PDE4 inhibitor treatment reversed the decreased ratio of Bcl‐2/BAX and inhibited the increased expression of PDE4D in the cerebral cortex and hippocampus of AD mice. Further, the PDE4 inhibitors reversed the decreased levels of cAMP and expression of phosphorylated cAMP response element‐binding protein (CREB) and brain derived neurotrophic factor (BDNF) in AD mice. These were mimicked by PDE4D deficiency and treatment with a PDE4D relatively selective inhibitor. In conclusions, these results suggest that PDE4 is an important target for the comorbidity of memory loss and depression in AD, which appears to be mediated by PDE4D‐cAMP signaling.Support or Funding InformationThis work was supported by research grants from NIH/NIA AG031687 and NIH/NIAAA HHSN275201700001C (both to HTZ) and the National Natural Science Foundation of China (81441111 to HW)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Ethanol Leaf Extract of Moringa Oleifera Blunts Isoproterenol‐induced Cardiotoxicity in Rats through Mitigation of Free Radical Production and Down Regulation of Cardiac Troponin and Nuclear Factor Kappa B
Authors: Adedapo Adeolu Alex , Asunloye Oluwatosin O , Adeoye Bisi Olajumoke , Oyagbemi Ademola A , Omobowale Temidayo O , Falayi Olufunke Olubunmi , Ogunpolu Blessing Seun , Oguntibeju Oluwafemi Omoniyi , Yakubu Momoh Audu 
Published: April 2020

Moringa oleifera is a plant that is widely distributed in the tropical environment and is used for medicinal purpose. It is believed that every part of this plant has medicinal value. The study evaluated the effect of the ethanol leaf extract of Moringa oleifera (EMOL) on isoproterenolinduced myocardial infarction in rats. Thirty‐five male Wistar rats weighing between 100 and 160 mg/kg were used for the study. The rats were randomly divided into 5 groups of 7 animals per group. Group A (control) animals were given normal saline, Group B (Toxicant) animals were given 100 mg/kg of isoproterenol administered subcutaneously on day 8. Groups C and D animals were given 50 mg/kg and 100 mg/kg of ethanol leaf extract of Moringa oleifera (ELMO) respectively for 7 days and on day 8 were given 100mg/kg of isoproterenol. Group E animals received 100 mg/kg of Moringa oleifera only for 7 days. On day 9 blood sample was taken for serum analysis of CK‐MB, Myeloperoxidase, Nitric oxide and Total protein, thereafter the animals were sacrificed via cervical dislocation and heart harvested for biochemical assays for the analysis of Advanced oxidative protein product (AOPP), Protein Carbonyl (PC), Malonaldehyde (MDA), Superoxide dismutase (SOD), Glutathione‐S‐Transferase (GST), Reduced glutathione (GSH), Glutathione peroxidase (GPx), Total thiol, Non‐protein thiol and Hydrogen peroxide (H2O2) generation), histopathology and immunohistochemistry (CTnI, NF‐KB).In this study there is significant (p < 0.05) reduction in SOD, GPx, GST, increased AOPP, PC, MDA, MPO levels and H2O2 generation in toxicant group (group B) but there was corresponding increase in the antioxidant enzymes and decrease in markers of oxidative stress and inflammation in ELMO treated groups. The toxicant also caused severe histopathological changes in the heart tissues but these were corrected by all the doses of the extract. There was increase expression of CTnI and NF‐KB in the toxicant group; pre‐treatment with ELMO however caused significant down regulation of all these proteins signifying its anti‐inflammatory and anti‐oxidative property.In conclusion, these experiments showed that ethanol leaf extract of Moringa oleifera possess cardiac protective effect through mitigation of free radicals production and down regulation of the studied cytokines.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Stretch‐induced activation of Hippo signaling in lung microvascular endothelial cells: A novel mechanism of overventilation‐induced pulmonary fibrosis
Authors: Turki Razan , Kapus Andras , Kuebler Wolfgang 
Published: April 2020

IntroductionMechanical ventilation is a mainstay of current therapy for patients with acute respiratory distress syndrome (ARDS). Yet despite its benefits, mechanical ventilation can also cause ventilator‐induced lung injury (VILI) that may ultimately progress to pulmonary fibrosis. In this study, we aimed to elucidate cellular mechanisms by which mechanical ventilation triggers pulmonary fibrosis. Specifically, we focused on the mechanosensitive transcriptional co‐factor TAZ, a key player in the Hippo‐signaling pathway involved in mechanosensation, cell proliferation, and profibrotic responses. As such, TAZ translocates to the nucleus of fibrotic lungs but not in healthy tissue. Interestingly, TAZ distribution upon cyclic stretch has not been extensively characterized and the underlying mechanisms are not well defined.ObjectivesTo examine the effect of mechanical stretch on the nuclear translocation of TAZ in endothelial cells, and to identify underlying regulatory mechanisms.MethodsTo mimic the effects of mechanical overventilation, human pulmonary microvascular endothelial cells (HPMECs) were subjected to stretch regimens at either 5%, mimicking low tidal volume ventilation, or 18% mimicking high tidal volume ventilation at 0.25 Hz for 1,3,6, 12h on a Flexcell 5000 Tension System and compared to static control (0%).ResultsMechanical stretch induced expression and release of the profibrotic cytokine TGF‐β1 from HPMECs and concomitantly caused translocation of TAZ from the cytosol to the nucleus in a dose‐ and time‐dependent manner. Specifically, the nuclear to cytoplasmic ratio (N/C) of TAZ increased after 3h of stretch at 18% compared to the static control whereas stretching these cells at 5% increased TAZ nuclear localization after 6h. Stretch‐induced TAZ translocation was not prevented by pharmacological inhibition of JNK or Src kinases, known regulators of the Hippo kinase LATS2; yet TAZ translocation was blocked by inhibition of Aurora kinases with SNS 314 mesylate (10UM). Consistently, inhibition of Aurora kinases decreased LATS2 phosphorylation at Ser83, a known modulation site of LATS2 activity.ConclusionThese data suggest that mechanical stretch activates TAZ and triggers release of pro‐fibrotic cytokines, at least partly, in an Aurora kinase and LATS2‐dependent mechanism. Interventions targeted at the Hippo‐signaling pathway as well as its upstream regulation may provide potential strategies to counteract the detrimental effects of mechanical ventilation.Support or Funding InformationThis work was supported by the Canadian Institutes of Health Research.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.