bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Novel Murine Knock‐in Model for Progranulin‐deficient Frontotemporal Dementia with Nonsense‐mediated mRNA Decay
Authors: Nguyen Andrew D. , Nguyen Thi A. , Zhang Jiasheng , Devireddy Swathi , Zhou Ping , Karydas Anna M. , Xu Xialian , Miller Bruce L. , Rigo Frank , Ferguson Shawn M. , Huang Eric J. , Walther Tobias C. , Farese Robert V. 
Published: June 2021

Frontotemporal dementia (FTD) is the most common neurodegenerative disorder in individuals under age 60 and has no treatment or cure. Since many cases of FTD result from GRN nonsense mutations, an animal model for this type of mutation is highly desirable for understanding pathogenesis and testing therapies. Here, we generated and characterized GrnR493X knock‐in mice, which model the most common human GRN mutation, a premature stop codon at arginine 493 (R493X). Homozygous GrnR493X mice have markedly reduced Grn mRNA levels, lack detectable progranulin protein, and phenocopy Grn knockout mice, with CNS microgliosis, reduced synaptic density, lipofuscinosis, hyper‐inflammatory macrophages, excessive grooming behavior, and reduced survival. Inhibition of nonsense‐mediated mRNA decay (NMD) by genetic, pharmacological, or antisense oligonucleotide‐based approaches showed that NMD contributes to the reduced mRNA levels in GrnR493X mice and cell lines and in fibroblasts from patients containing the GRNR493X mutation. The expressed truncated R493X mutant protein was functional in several assays in progranulin‐deficient cells, suggesting that NMD inhibition could be an effective as a therapeutic approach for treating progranulin deficiency caused by the R493X mutation. The GrnR493X mice and associated cell lines provide novel model systems to identify therapies for treating FTD resulting from GRN nonsense mutations.Support or Funding InformationThis work is supported by National Institutes of Health Grants AG023501 (to R.V.F.) and AG047339 (to A.D.N.), VA Merit Award BX002978 (to E.J.H.), the Consortium for Frontotemporal Dementia Research (to R.V.F., T.C.W., E.J.H., S.M.F., and B.L.M.), and the Gladstone Institutes. T.C.W. is an investigator of the Howard Hughes Medical Institute. The Gladstone Institutes received support from a National Center for Research Resources grant RR18928.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Discovery, Efficacy Testing, and Potential Target Identification of Antibiofilm Compounds
Authors: Goode David , Jackson Caroline , Aber Kayla , Shah Ashna , Hensel Linda 
Published: June 2021

Ideal biofilm inhibitors would not affect the growth of bacteria, in general, but would be specific for inhibiting reception of the signals to initiate biofilm formation. In this manner, the inhibitor would not affect the growth of normal flora that are imperative for the health of the host. Compounds that resemble biofilm signaling molecules have the potential to competitively inhibit the receptors from receiving signals. Over 200 novel compounds were synthesized and tested by undergraduate students in a sophomore integrated biology and organic chemistry classroom setting. Compounds that demonstrated a statistically significant inhibition of biofilm formation and that had no bacteriostatic or bactericidal activity in one or more of three species of bacteria (Bacillus subtilus, Staphylococcus aureus, and Pseudomonas aeruginosa) in the hands of the undergraduates underwent further testing. Biofilm inhibition and enhancement was confirmed with a modified crystal violet assay, and growth curve analysis confirmed previous disk diffusion and use‐dilution assay results. Hit compounds against Pseudomonas aeruginosa were further tested in a genetics undergraduate classroom using an engineered quorum sensing modulation assay. These results were confirmed in the research laboratory, which along with computational modeling point to LasR as the possible protein target of the novel compounds in Pseudomonas aeruginosa.Support or Funding InformationThis research is supported by the Mercer University Quality Enhancement Plan: Research that Reaches Out.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Isolation and Characterization of Antimicrobial Compound Isolated from Lichen Parmelia vagans.
Authors: Bondarenko Vladimir 
Published: June 2021

This study was aimed to isolate and characterize the antibacterial compounds from lichen Parmelia vagans.Many pharmaceutical products today have derivatives from living organisms. Recently, because of the deficiency of new antibiotics, we once again turned to the nature as a source of new antibacterial compounds. Lichens have been found to have secondary metabolites which have been found to have various physiological effects including antimicrobial activity. A lichen is a composite organism composed of a photobiont such as an alga or cyanobacterium and a mycobiont, a fungus. In this study, the antimicrobial activity of extracts from a lichen identified as Parmelia vagans was evaluated.Extracts of Parmelia vagans were initially obtained by using various solvents to find the most efficient extraction of antimicrobial compound(s). We separated a crude ethyl acetate lichen extract using reverse phase chromatography and analyzed the fraction using the Resazurin cell viability assay. We used Staphylococcus aureus in the assay because as it has been shown previously, it has relatively high sensitivity to the antibacterial activity of the lichen extract. We identified two major peaks of antimicrobial activity, one peak contained usnic acid, a well‐characterized antibacterial compound, and the second peak contained a novel compound that usually co‐migrates with usnic acid on TLC plate. It was observed that newly isolated compound had no inhibitory effect on the growth of Escherichia coli, and Klebsiella pneumonia at the tested concentrations, but inhibition of growth was observed in Staphylococcus aureus, Bacillus subtilis, and Streptococcus pneumoniae.Using reverse phase chromatography we isolated a novel antimicrobial compound that will be further characterized structurally and functionally. It will be tested against various gram‐positive and gram‐negative bacterial strains to identify its bacterial target(s).Support or Funding InformationThe above research project was supported by Touro University Nevada intramural research grant.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Unconventional Secretion of Adipocyte Fatty Acid Binding Protein (FABP4) by Adipocytes
Authors: Josephrajan Ajeetha , Hertzel Ann V. , Mashek Douglas , Kim Do‐Hyung , Imai Shin‐Ichiro , Bernlohr David A. 
Published: June 2021

Fatty acid binding protein 4 (FABP4) is a cytoplasmic lipid carrier functioning in adipocytes to facilitate lipolysis. Genetic or pharmacologic depletion of the protein provides protection from insulin resistance, asthma, atherosclerosis and inflammation. Metabolic dysfunction has been considered due to the intracellular functions of the protein. However, recent literature has shown that serum FABP4 levels are elevated in patients with metabolic syndrome. FABP4 is a leaderless protein and the mechanism of its release from adipocytes is unknown. Although FABP4 secretion increased with increasing intracellular free fatty acid (FFA) levels and correlated positively with FFA efflux, the molar amount of FABP4 secreted is less than 1% of total molar FFA efflux implying that the protein does not mediate release of bulk lipid in response to lipolytic stimulation. To evaluate the mechanism of FABP4 secretion, we have used primary mouse adipocytes and the 3T3‐L1 adipocyte cell culture model. Results from inhibitor/activator studies showed requirement for some, but not all components of autophagy. For example, classical autophagic inhibitors such as 3‐methyl adenine or the Class III PI 3‐Kinase VPS34 specific inhibitor attenuated FABP4 secretion, while autophagic inducer N‐acetyl sphingosine induced FABP4 secretion. Moreover, FABP4 secretion was potentiated when lipophagy and lysosomal exocytosis were blocked using chemical inhibitors of lysosomal acid lipase or Bafilomycin A1, respectively. Consistent with these results, genetic knockdown of ATG13 or FIP200 (involved in early events of phagophore formation) attenuated secretion, whereas silencing of Atg5 had little effect on FABP4 secretion. Similar to the adipocyte secretion of eNAMPT, blocking SIRT1 by the chemical inhibitor EX527 attenuated secretion, while activating SIRT1 by resveratrol potentiated secretion. Our results show that FABP4 is secreted through autophagy mediated alternative secretion and regulated in part by SIRT1.Support or Funding InformationSupported by NIH DK053189 to DAB.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differentiation of spermatogonial stem cell‐derived embryonic stem (SSC‐ES) cells towards retinal ganglion cells
Authors: Lee Tin‐Lap , Suen Hoi Ching Ashley , Qian Yan , Liao Jinyue , Luk Chun Shui Alfred , Chan Wai Yee , Ng Tsz Kin Michael , Leung Kai Shun Christopher 
Published: June 2021

Embryonic stem cells (ESCs) have a wide range of clinical applications. However, its ethical concerns cannot be neglected. In this study, we collected spermatogonial stem cells (SSCs) from mouse testes and they were subsequently transdifferentiated to ESC‐like cells through a non‐viral and non‐invasive approach. These SSC‐derived ESC‐like cells exhibited stem cell properties similar to ESCs in terms of morphology and pluripotency gene expression profile. Next, we aimed to generate retinal ganglion cells using spermatogonial stem cell‐derived embryonic stem (SSC‐ES) cells. Following previously established protocols with several modifications, SSC‐ES cells were differentiated towards retinal cell lineage and developed optic vesicles in 7–10 days in in vitro culture, while some cells generated neurites in 13–16 days. These neuron‐like cells were positive for RGC‐specific marker Brn3b and other neural markers, thus confirming their RGC identity. We will compare the regenerative effect in vivo using transplantation assay. These cell populations could provide a novel cell‐based therapeutic option for retinal degenerative diseases.Support or Funding InformationHealth and Medical Research Fund, Food and Health Bureau. Hong Kong SAR Government.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Conformational and Thermodynamic Features of Meibum in Adolescents and Adults with Graft‐versus‐host Disease
Authors: Sledge Samiyyah M. , Blackburn Ryan , Ramasubramanian Aparna , Borchman Douglas , Yappert Marta C. 
Published: June 2021

PurposeGraft‐versus‐host disease (GvHD) is a common complication of allogenic hematological stem cell transplantations that is often associated with dry eye disease (DED) classified as a mixture of meibomian gland dysfunction and aqueous tear deficiency. The temperature‐dependence of conformational changes in human meibum (hM) was measured in a comparative study to elucidate the potential impact of GvHD on hM composition, structure and function with DED.MethodsDonors received full ophthalmic eye exams and DED was clinically diagnosed. A total of 28 donors without DED were separated into two pools: adolescent (age: 17 ± 1 and 63% male) and adult (age: 42 ± 4 and 73% male). Adolescent (age: 15 ± 1 and 80% male) and adult (age: 45 ± 5 and 30% male) pools from a total of 15 donors affected by both DED and GvHD were also tested. The extracted hM was placed in d‐chloroform and analyzed by Fourier Transform Infrared Spectroscopy (FTIR).ResultsAmong the measured parameters, the lipid order (% trans) in hM at 36°C increased significantly from adolescents without DED (35.3 ± 4%) to those affected by GvHD (54.7 ± 4%). A similar trend (33.3 ± 0.09% without DED to 38.0 ± 4% with DED/GvHD) was observed in hM from adults. The change in enthalpy for the order‐to‐disorder transition decreased significantly with DED/GvHD in both adolescents (from 163 ± 10 Kcal/mol without DED to 134 ± 9 Kcal/mol with DED/GvHD) and adults (141 ± 5 Kcal/mol without DED to 126 ± 11 Kcal/mol with DED/GvHD). The phase transition temperature (C°) was significantly higher in adolescents with DED/GvHD (38 ± 1°C) compared to donors without DED (30.4 ± 0.6°C), but the increase was less pronounced in adults with DED/GvHD (31 ± 1°C) relative to those without DED (30.0 ± 0.4°C).ConclusionBoth age and DED lead to conformational and thermodynamic differences in hM. Increases in lipid order and transition temperatures were observed in hM with GvHD. However, these relative increases were more pronounced in hM from adolescents than from adults. The decrease of change in enthalpy for adolescents was more pronounced compared with adults. The conformational and thermodynamic differences observed with age and, importantly, with DED/GvHD are indicative of compositional changes in hM that could impact tear film stability.Support or Funding InformationMajor support was obtained from the National Institute of Health (NIH) EYO RO126180 (DB); NIH Research Supplements to Promote Diversity in Health‐Related Research EYO RO126180S‐1 (SS); and an unrestricted grant from Research to Prevent Blindness, Inc. New York, NY, USA.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anatomical Findings of fetuses Vertically‐infected with Zika Virus
Authors: Figueredo Luisa Fernanda , Franco‐Zuluaga Jorge Andres , Carrère‐Rivera Camille , Rodriguez‐Silva Daniela Sofia , Akle Veronica 
Published: June 2021

Zika virus (ZIKV) is a new TORCHS virus with a recent outbreak in Latin America and south of the United States. The virus can be vertically‐transmitted, with many implications for the placenta and the fetuses. Even though there are multiple cases of malformations in babies born from mothers infected with ZIKV, there is little detail about the effect of the infection in the tissues during development.In this study, four fetuses of the second and third trimester and a newborn, who died within few weeks, from mothers diagnosed with ZIKV were collected from The Legal Medicine Institute of Bogota in Colombia. The cases were subjected to a complete autopsy and the necropsy results were analyzed. For histologic examination, series of micro‐sections were stained with H&E or processed for immunostaining.Macroscopic findings included compromise of the placenta characterized by opacity and thickness of membranes. The fetuses presented severe edema in extremities and pulmonary hypoplasia. In addition, one case showed Potter facies and low implantation of ear and nose. The newborn also presented petechiae on the thoracic region. Microscopic examination showed severe endothelial lesions in ovular membranes associated with inflammation and dystrophic calcifications of maternal and fetal sides of the placenta. Additionally, there were signs of severe funisitis in the umbilical cord, and congestion of the lungs and accumulation of plasmocytes in the fetuses.Preliminary results suggest an association between damage of the placental barrier and facilitation of fetal infection. Also, there is evidence of inflammation throughout the fetus' and newborn's tissues, in all cases. A through examination of all organs would give a better overview of the morphological changes caused by the viral infection, which could help understand the infection and give insights into the mechanisms of transmission, prevention and future treatment strategies.Support or Funding InformationSchool of Medicine. Universidad de los Andes. Bogotá ‐ ColombiaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
EFFECT OF STAPHYLOCOCCAL ENTEROTOXINS TYPE A (SEA) AND B (SEB) ON THE NEUTROPHIL DYSFUNCTION DURING STAPHYLOCOCCUS AUREUSINDUCED SEPSIS
Authors: Duarte Ana Paula Ferreira , Pinheiro‐Torres Anelize Sgorlon , de Souza Ivani Aparecida , Antunes Edson 
Published: June 2021

Sepsis is a systemic inflammatory disease which affects about 17 million people per year worldwide, with a mortality rate above 50%. Sepsis by gram‐positive bacteria has a higher mortality rate when compared with sepsis by gram‐negative bacteria. However, experimental models that adequately mimic the signs of sepsis by gram‐positive bacteria are limited. An inefficient neutrophils (NE) mobilization in Staphylococcus aureus‐induced sepsis is the main cause of death in patients infected with this bacterium. Staphylococcus aureus pathological effects are strong related to the secretion of staphylococcal enterotoxins (SEs). Recently, we have shown that human eosinophils and mice bone marrow granulocytes incubated with SEs exhibit reduced in vitro chemotaxis and adhesion activity. The present study aims to identify the effects of SEA and SEB on the functional properties of human NE. Blood was collected from healthy volunteers after approval from the local ethics committee (Protocol No 61370616.3.0000.5412). Blood samples were placed on isotonic Percoll solution. After centrifugation, the mononuclear cell layer was removed and the pellet containing erythrocytes and granulocytes was aspirated and subjected to isotonic lysis. After lysis cells (98% NE) were resuspended to 4 × 106 cells/ml and submitted to in vitro incubation with SEA or SEB (100 ng/ml; 2 h). Adhesion assays were carried out in 96‐well plates pre‐coated with recombinant human VCAM‐1 and ICAM‐1 for 30 min in the presence of interleukin‐8 (IL‐8). The NE adhesion was calculated by measuring myeloperoxidase (MPO) activity on adherent cells. NE migration assay was performed using a 48‐well microchemotaxis Boyden chamber. Both SEA and SEB reduces human neutrophil adhesion in VCAM‐1 (Control: 4.7 ± 0.5; untreated NE + IL‐8: 6.7 ± 0.6; SEA treated NE+IL‐8: 4.9 ± 0.8; SEB treated NE + IL‐8: 3.7 ± 0.6*# optic density/NE × 106 cells; ) or ICAM‐1(Control: 4.3 ± 0.5; untreated NE + IL‐8: 6.1 ± 0.6; SEA treated NE+IL‐8: 5.5 ± 1.0; SEB treated NE + IL‐8: 3.7 ± 0.9 optic density/NE × 106 cells) coated plates. Similar results were observed for NE chemotactic activity (Control: 18.7 ± 0.8; untreated NE +IL‐8: 52.9 ± 2.7; SEA treated NE+IL‐8: 28.5 ± 1.4; SEB treated NE + IL‐8: 34.2 ± 1.4 NE per high‐power field). The inhibitory effect of SEA and SEB on human NE in vitro adhesion and chemotaxis suggests a role of these toxins on the neutrophil dysfunction associated with the severe sepsis by Staphylococcus aureus.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
INVOLVEMENT OF TLR4 AND PPAR‐α RECEPTORS IN HOST RESPONSE AND NLRP3 INFLAMMOSOME ACTIVATION, AGAINST PULMONARY INFECTION WITH PSEUDOMOSAS AERUGINOSA
Authors: GUGLIANDOLO ENRICO , Fusco Roberta , Ginestra Giovanna , D'amico Ramona , Bisignano Carlo , Mandalari Giuseppina , Cuzzocrea Salvatore , Di Paola Rosanna 
Published: June 2021

BackgroundColonization with Pseudomonas aeruginosa (PA), the most common pathogen isolated mainly in patients with cystic fibrosis, is particularly difficult to eradicate and is associated with acceleration of decline in lung function and with poorer prognosis. Host resistance to PA is dependent on the activation of an innate immune response. The initiation of this response requires the recognition of bacterial ligands by signaling receptors. Recently, Toll‐like receptors (TLRs) have been found to have prominent roles in the activation of innate immune responses to infection. PA LPS is recognized by TLR4 and has been shown to induce lung inflammation in vivo. In addition, regulation of this process is essential for proper pathogen clearance and to prevent excessive inflammatory response resulting in tissue damage. This occurs through the production of different cytokines, and the activation of different pathways such as inflammasome activation, which is responsible for the production of IL‐1b. Moreover the airway epithelial cells respond with increased production of proinflammatory cytokines as a result of increased NF‐κB activation. One potential regulator of NF‐κB activation are the peroxisome proliferator‐activated receptors (PPARs), and PPARa is a member of the ligand‐activated nuclear receptor superfamily. Thus, the purpose of the present study was to evaluate the role of TLR4 and PPARa in the pulmonary innate immunity response to PA and in the consequent inflammatory response.MethodsTo evaluate the involvement of TLR4 and PPARa in a PA infection, we used TLR4 KO and PPARa KO mice who received an intratracheal (i.t.) administration of 50 ml of PA strain (106 CFU), thus evaluating if these mice were profoundly susceptible to PA compared to WT mice.ResultsThe results of the present study showed that administration of PA worsened the pathophysiology of PA lung disease in TLR4 and PPARa KO mice compared to WT.ConclusionsThe present study demonstrated that TLR4 and PPARa receptors would mediate the earliest control of bacterial replication as well as proinflammatory responses to PA infections, and in particular that PPARa receptors are needed to prevent an excessive inflammatory response, as in the control of the inflammosome complex NLP3 activation.Support or Funding Informationno fundingThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.