{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:51:03Z","timestamp":1760147463906,"version":"build-2065373602"},"reference-count":50,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2023,2,4]],"date-time":"2023-02-04T00:00:00Z","timestamp":1675468800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006315","name":"Dutch Arthritis Society","doi-asserted-by":"publisher","award":["17-1-403"],"award-info":[{"award-number":["17-1-403"]}],"id":[{"id":"10.13039\/501100006315","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Spondyloarthritis (SpA) patients suffer from joint inflammation resulting in tissue damage, characterized by the presence of numerous neutrophils in the synovium and synovial fluid (SF). As it is yet unclear to what extent neutrophils contribute to the pathogenesis of SpA, we set out to study SF neutrophils in more detail. We analyzed the functionality of SF neutrophils of 20 SpA patients and 7 disease controls, determining ROS production and degranulation in response to various stimuli. In addition, the effect of SF on neutrophil function was determined. Surprisingly, our data show that SF neutrophils in SpA patients have an inactive phenotype, despite the presence of many neutrophil-activating stimuli such as GM-CSF and TNF in SF. This was not due to exhaustion as SF neutrophils readily responded to stimulation. Therefore, this finding suggests that one or more inhibitors of neutrophil activation may be present in SF. Indeed, when blood neutrophils from healthy donors were activated in the presence of increasing concentrations of SF from SpA patients, degranulation and ROS production were dose-dependently inhibited. This effect was independent of diagnosis, gender, age, and medication in the patients from which the SF was isolated. Treatment of SF with the enzyme hyaluronidase strongly reduced the inhibitory effect of SF on neutrophil activation, indicating that hyaluronic acid that is present in SF may be an important factor in preventing SF neutrophil activation. This finding provides novel insights into the role of soluble factors in SF regulating neutrophil function and may lead to the development of novel therapeutics targeting neutrophil activation via hyaluronic acid or associated pathways.<\/jats:p>","DOI":"10.3390\/ijms24043066","type":"journal-article","created":{"date-parts":[[2023,2,6]],"date-time":"2023-02-06T02:57:32Z","timestamp":1675652252000},"page":"3066","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Hyaluronic Acid in Synovial Fluid Prevents Neutrophil Activation in Spondyloarthritis"],"prefix":"10.3390","volume":"24","author":[{"given":"Sanne","family":"Mol","sequence":"first","affiliation":[{"name":"Department of Experimental Immunology, Amsterdam Institute for Infection and Immunity, Amsterdam UMC, Location AMC, 1105 AZ Amsterdam, The Netherlands"},{"name":"Department of Biomolecular Health Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584 CL Utrecht, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Esther W. M.","family":"Taanman-Kueter","sequence":"additional","affiliation":[{"name":"Department of Experimental Immunology, Amsterdam Institute for Infection and Immunity, Amsterdam UMC, Location AMC, 1105 AZ Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Baltus A.","family":"van der Steen","sequence":"additional","affiliation":[{"name":"Department of Experimental Immunology, Amsterdam Institute for Infection and Immunity, Amsterdam UMC, Location AMC, 1105 AZ Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tom","family":"Groot Kormelink","sequence":"additional","affiliation":[{"name":"Department of Experimental Immunology, Amsterdam Institute for Infection and Immunity, Amsterdam UMC, Location AMC, 1105 AZ Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marleen G. H.","family":"van de Sande","sequence":"additional","affiliation":[{"name":"Department of Experimental Immunology, Amsterdam Institute for Infection and Immunity, Amsterdam UMC, Location AMC, 1105 AZ Amsterdam, The Netherlands"},{"name":"Department of Rheumatology and Clinical Immunology, Amsterdam Rheumatology and Immunology Center, Amsterdam University Medical Centers, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8940-1788","authenticated-orcid":false,"given":"Sander W.","family":"Tas","sequence":"additional","affiliation":[{"name":"Department of Experimental Immunology, Amsterdam Institute for Infection and Immunity, Amsterdam UMC, Location AMC, 1105 AZ Amsterdam, The Netherlands"},{"name":"Department of Rheumatology and Clinical Immunology, Amsterdam Rheumatology and Immunology Center, Amsterdam University Medical Centers, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0360-0311","authenticated-orcid":false,"given":"Marca H. M.","family":"Wauben","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Health Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584 CL Utrecht, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Esther C.","family":"de Jong","sequence":"additional","affiliation":[{"name":"Department of Experimental Immunology, Amsterdam Institute for Infection and Immunity, Amsterdam UMC, Location AMC, 1105 AZ Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Brennan-Olsen, S.L., Cook, S., Leech, M.T., Bowe, S.J., Kowal, P., Naidoo, N., Ackerman, I.N., Page, R.S., Hosking, S.M., and Pasco, J.A. (2017). Prevalence of Arthritis According to Age, Sex and Socioeconomic Status in Six Low and Middle Income Countries: Analysis of Data from the World Health Organization Study on Global AGEing and Adult Health (SAGE) Wave 1. BMC Musculoskelet. Disord., 18.","DOI":"10.1186\/s12891-017-1624-z"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1320","DOI":"10.1002\/acr.22831","article-title":"Global Prevalence of Spondyloarthritis: A Systematic Review and Meta-Regression Analysis","volume":"68","author":"Stolwijk","year":"2016","journal-title":"Arthritis Care Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1097\/MAJ.0b013e318294457f","article-title":"The Epidemiology of Back Pain, Axial Spondyloarthritis and HLA-B27 in the United States","volume":"345","author":"Reveille","year":"2013","journal-title":"Am. J. Med. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"211","DOI":"10.3899\/jrheum.181331","article-title":"Peripheral Manifestations in Spondyloarthritis and Their Effect: An Ancillary Analysis of the ASAS-COMOSPA Study","volume":"47","author":"Dougados","year":"2020","journal-title":"J. Rheumatol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1007\/s11926-013-0316-4","article-title":"Psoriatic Arthritis: Phenotypic Variance and Nosology","volume":"15","author":"Eder","year":"2013","journal-title":"Curr. Rheumatol. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"vi23","DOI":"10.1093\/rheumatology\/key152","article-title":"Therapies in Ankylosing Spondylitis-from Clinical Trials to Clinical Practice","volume":"57","author":"Tahir","year":"2018","journal-title":"Rheumatology"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1136\/annrheumdis-2020-218304","article-title":"Immune Response Profiling of Patients with Spondyloarthritis Reveals Signalling Networks Mediating TNF-Blocker Function in Vivo","volume":"80","author":"Menegatti","year":"2021","journal-title":"Ann. Rheum. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1186\/s13075-017-1249-5","article-title":"Mechanistic Rationales for Targeting Interleukin-17A in Spondyloarthritis","volume":"19","author":"Raychaudhuri","year":"2017","journal-title":"Arthritis Res. Ther."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"51","DOI":"10.3389\/fimmu.2018.02456","article-title":"Understanding the Multifaceted Role of Neutrophils in Cancer and Autoimmune Diseases","volume":"9","author":"Wang","year":"2018","journal-title":"Front. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1146\/annurev-pathol-020712-164023","article-title":"The Multifaceted Functions of Neutrophils","volume":"9","author":"Mayadas","year":"2013","journal-title":"Annu. Rev. Pathol. Mech. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1146\/annurev-biochem-060815-014442","article-title":"Reactive Oxygen Species and Neutrophil Function","volume":"85","author":"Winterbourn","year":"2016","journal-title":"Annu. Rev. Biochem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1438","DOI":"10.1023\/B:DDAS.0000042243.47279.87","article-title":"Relationship between Fecal Calprotectin, Intestinal Inflammation, and Peripheral Blood Neutrophils in Patients with Active Ulcerative Colitis","volume":"49","author":"Hanai","year":"2004","journal-title":"Dig. Dis. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1002\/jcla.21908","article-title":"The Association Between Neutrophil\/Lymphocyte Ratio and Disease Activity in Rheumatoid Arthritis and Ankylosing Spondylitis","volume":"30","author":"Mercan","year":"2016","journal-title":"J. Clin. Lab. Anal."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1186\/s13075-021-02693-7","article-title":"A Role for Neutrophils in Early Enthesitis in Spondyloarthritis","volume":"24","author":"Stavre","year":"2022","journal-title":"Arthritis Res. Ther."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1007\/BF03326409","article-title":"Possible Involvement of Neutrophil Elastase in Impaired Mucosal Repair in Patients with Ulcerative Colitis","volume":"37","author":"Kuno","year":"2002","journal-title":"J. Gastroenterol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2376","DOI":"10.3389\/fimmu.2019.02376","article-title":"Neutrophils in Psoriasis","volume":"10","author":"Chiang","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3709749","DOI":"10.1155\/2020\/3709749","article-title":"Role of Neutrophils in Psoriasis","volume":"2020","author":"Wang","year":"2020","journal-title":"J. Immunol. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"584116","DOI":"10.3389\/fimmu.2020.584116","article-title":"Rheumatoid Arthritis Synovial Fluid Neutrophils Drive Inflammation through Production of Chemokines, Reactive Oxygen Species, and Neutrophil Extracellular Traps","volume":"11","author":"Wright","year":"2021","journal-title":"Front. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1007\/s10067-010-1646-5","article-title":"Th-17 Associated Cytokines in Patients with Reactive Arthritis\/ Undifferentiated Spondyloarthropathy","volume":"30","author":"Singh","year":"2011","journal-title":"Clin. Rheumatol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2286","DOI":"10.1016\/j.jaci.2018.01.003","article-title":"Human TH17 Cell Development Requires Processing of Dendritic Cell\u2013Derived CXCL8 by Neutrophil Elastase","volume":"141","author":"Souwer","year":"2018","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"340","DOI":"10.1016\/j.it.2007.06.002","article-title":"Neutrophil Granules: A Library of Innate Immunity Proteins","volume":"28","author":"Borregaard","year":"2007","journal-title":"Trends Immunol."},{"key":"ref_22","unstructured":"(2006). Paige Lacy Mechanisms of Degranulation in Neutrophils. Allergy Asthma Clin. Immunol., 2, 98\u2013108."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Mol, S., Hafkamp, F.M.J., Varela, L., Simkhada, N., Taanman-Kueter, E.W., Tas, S.W., Wauben, M.H.M., Kormelink, T.G., and de Jong, E.C. (2021). Efficient Neutrophil Activation Requires Two Simultaneous Activating Stimuli. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms221810106"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1189\/jlb.1212619","article-title":"Proteome Profiling of Human Neutrophil Granule Subsets, Secretory Vesicles, and Cell Membrane: Correlation with Transcriptome Profiling of Neutrophil Precursors","volume":"94","author":"Heegaard","year":"2013","journal-title":"J. Leukoc. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1016\/j.bbamcr.2012.11.027","article-title":"ADAM17 Cleaves CD16b (Fc\u03b3RIIIb) in Human Neutrophils","volume":"1833","author":"Wang","year":"2013","journal-title":"Biochim. Biophys Acta Mol. Cell Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1136\/ard.44.12.817","article-title":"Concentration and Molecular Weight of Sodium Hyaluronate in Synovial Fluid from Patients with Rheumatoid Arthritis and Other Arthropathies","volume":"44","author":"Dahl","year":"1985","journal-title":"Ann. Rheum. Dis."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"4","DOI":"10.3389\/fimmu.2015.00231","article-title":"Hyaluronan\u2014A Functional and Structural Sweet Spot in the Tissue Microenvironment","volume":"6","author":"Monslow","year":"2015","journal-title":"Front. Immunol"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1007\/BF02005763","article-title":"Hyaluronic Acid Modulates Acute and Chronic Inflammation","volume":"43","author":"Ialenti","year":"1994","journal-title":"Agents Actions"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1303","DOI":"10.1016\/0092-8674(90)90694-A","article-title":"CD44 Is the Principal Cell Surface Receptor for Hyaluronate","volume":"61","author":"Aruffo","year":"1990","journal-title":"Cell"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1016\/S0021-9258(18)64363-7","article-title":"The Mechanism of Action of Hyaluronidase","volume":"236","author":"Ludowieg","year":"1961","journal-title":"J. Biol. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"465","DOI":"10.3904\/kjim.2018.364","article-title":"Effect of Biologics in the Level of Cytokines in the Synovial Fluid of Patients with Ankylosing Spondylitis","volume":"35","author":"Koo","year":"2020","journal-title":"Korean J. Intern. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1136\/ard.51.6.731","article-title":"Determination of Cytokines in Synovial Fluids: Correlation with Diagnosis and Histomorphological Characteristics of Synovial Tissue","volume":"51","author":"Kahle","year":"1992","journal-title":"Ann. Rheum. Dis."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1007\/BF00262295","article-title":"Measurement of Colony-Stimulating Factors in Synovial Fluid: Potential Clinical Value","volume":"14","author":"Bell","year":"1995","journal-title":"Rheumatol. Int."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1046\/j.1365-2567.1997.00249.x","article-title":"Neutrophils from the Synovial Fluid of Patients with Rheumatoid Arthritis Express the High Affinity Immunoglobulin G Receptor, Fc\u03b3R1 (CD64): Role of Immune Complexes and Cytokines in Induction of Receptor Expression","volume":"91","author":"Quayle","year":"1997","journal-title":"Immunology"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"111","DOI":"10.2478\/intox-2013-0019","article-title":"Hyaluronan and Synovial Joint: Function, Distribution and Healing","volume":"6","author":"Tamer","year":"2013","journal-title":"Interdiscip. Toxicol."},{"key":"ref_36","first-page":"78","article-title":"Hyaluronic Acid in Inflammation and Tissue Regeneration","volume":"28","author":"Litwiniuk","year":"2016","journal-title":"Wounds"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1113\/jphysiol.2005.088906","article-title":"Size Selectivity of Hyaluronan Molecular Sieving by Extracellular Matrix in Rabbit Synovial Joints","volume":"567","author":"Sabaratnam","year":"2005","journal-title":"J. Physiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1136\/ard.44.10.647","article-title":"Hyaluronic Acid Production in Vitro by Synovial Lining Cells from Normal and Rheumatoid Joints","volume":"44","author":"Dahl","year":"1985","journal-title":"Ann. Rheum. Dis."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1002\/art.1780310110","article-title":"Antioxidant Activity of Synovial Fluid, Hyaluronic Acid, and Two Subcomponents of Hyaluronic Acid. Synovial Fluid Scavenging Effect Is Enhanced in Rheumatoid Arthritis Patients","volume":"31","author":"Sato","year":"1988","journal-title":"Arthritis Rheum."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"181","DOI":"10.3109\/03008209309016825","article-title":"Inhibitory Effects of Hyaluronan on Neutrophil-Mediated Cartilage Degradation","volume":"29","author":"Tobetto","year":"1993","journal-title":"Connect Tissue Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"875","DOI":"10.1002\/art.41605","article-title":"Synovial Fluid Neutrophils From Patients With Juvenile Idiopathic Arthritis Display a Hyperactivated Phenotype","volume":"73","author":"Metzemaekers","year":"2021","journal-title":"Arthritis Rheumatol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1186\/s13075-021-02483-1","article-title":"Synovial Fluid Neutrophils in Oligoarticular Juvenile Idiopathic Arthritis Have an Altered Phenotype and Impaired Effector Functions","volume":"23","author":"Schmidt","year":"2021","journal-title":"Arthritis Res. Ther."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"R122","DOI":"10.1186\/ar4579","article-title":"Enhanced Neutrophil Extracellular Trap Generation in Rheumatoid Arthritis: Analysis of Underlying Signal Transduction Pathways and Potential Diagnostic Utility","volume":"16","author":"Giaglis","year":"2014","journal-title":"Arthritis Res. Ther."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1136\/ard.50.4.237","article-title":"Activation of the Neutrophil Myeloperoxidase-H2O2 System by Synovial Fluid Isolated from Patients with Rheumatoid Arthritis","volume":"50","author":"Nurcombe","year":"1991","journal-title":"Ann. Rheum. Dis."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1042\/bj2860345","article-title":"Activation of Neutrophil Reactive-Oxidant Production by Synovial Fluid from Patients with Inflammatory Joint Disease: Soluble and Insoluble Immunoglobulin Aggregates Activate Different Pathways in Primed and Unprimed Cells","volume":"286","author":"Robinson","year":"1992","journal-title":"Biochem. J."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"910","DOI":"10.1136\/ard.54.11.910","article-title":"Human Blood and Synovial Fluid Neutrophils Cultured in Vitro Undergo Programmed Cell Death Which Is Promoted by the Addition of Synovial Fluid","volume":"54","author":"Bell","year":"1995","journal-title":"Ann. Rheum. Dis."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1249","DOI":"10.1093\/rheumatology\/41.11.1249","article-title":"Synovial Fluid from Patients with Rheumatoid Arthritis Inhibits Neutrophil Apoptosis: Role of Adenosine and Proinflammatory Cytokines","volume":"41","author":"Ottonello","year":"2002","journal-title":"Rheumatology"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"25549","DOI":"10.52965\/001c.25549","article-title":"A Comprehensive Review of Viscosupplementation in Osteoarthritis of the Knee","volume":"13","author":"Peck","year":"2021","journal-title":"Orthop. Rev."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1016\/j.semarthrit.2015.04.011","article-title":"Consensus Statement on Viscosupplementation with Hyaluronic Acid for the Management of Osteoarthritis","volume":"45","author":"Henrotin","year":"2015","journal-title":"Semin. Arthritis Rheum."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"973","DOI":"10.1080\/14397595.2016.1270496","article-title":"Short-Term Effect of Ultrasound-Guided Low-Molecular-Weight Hyaluronic Acid Injection on Clinical Outcomes and Imaging Changes in Patients with Rheumatoid Arthritis of the Ankle and Foot Joints. A Randomized Controlled Pilot Trial","volume":"27","author":"Wang","year":"2017","journal-title":"Mod. Rheumatol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/4\/3066\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:24:05Z","timestamp":1760120645000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/4\/3066"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,2,4]]},"references-count":50,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2023,2]]}},"alternative-id":["ijms24043066"],"URL":"https:\/\/doi.org\/10.3390\/ijms24043066","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2023,2,4]]}}}