{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T11:24:27Z","timestamp":1791804267251,"version":"4.3.4"},"reference-count":68,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T00:00:00Z","timestamp":1753401600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Dement."],"abstract":"<jats:p>Alzheimer\u2019s disease (AD) is a severe neurodegenerative brain disorder molecularly characterized by extracellular \u03b2-amyloid plaques, intraneuronal tau neurofibrillary tangles, cholinergic neuron death, neuroinflammation, vascular damage, and astroglial and microglial activation. AD is a complex disorder, with &amp;gt;99% of all cases being sporadic and typically occuring around the age of 65. Due to this intricate nature of the disorder, <jats:italic>in vitro<\/jats:italic> experiments have limitations; however, three-dimensional organotypic brain slices may offer the best alternative for studying the mechanisms involved in the progression of AD. This review provides an overview of how to study the general aspects of AD <jats:italic>ex vivo<\/jats:italic>, focusing on (a) \u03b2-amyloid plaques in brain slices, (b) tau pathology induced by chemical drugs, (c) cell death of cholinergic neurons and protection by nerve growth factor, (d) activation of astrocytes and microglia, and (e) vascular pathologies, including the role of platelets. Furthermore, we investigated (f) how microcontact printing on brain slices can be used to study the spread of \u03b2-amyloid and tau, and (g) how brain slices can help identify novel human AD biomarkers.<\/jats:p>","DOI":"10.3389\/frdem.2025.1585124","type":"journal-article","created":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T06:13:09Z","timestamp":1753423989000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Three-dimensional organotypic mouse brain slices to study Alzheimer\u2019s disease pathologies: a review"],"prefix":"10.3389","volume":"4","author":[{"given":"Christian","family":"Humpel","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,7,25]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1007\/s40265-015-0529-0","article-title":"Tau biology and tau-directed therapies for Alzheimer's disease","volume":"76","author":"Bakota","year":"2016","journal-title":"Drugs"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"105192","DOI":"10.1016\/j.neubiorev.2023.105192","article-title":"New perspectives on the basal forebrain cholinergic system in Alzheimer's disease","volume":"150","author":"Berry","year":"2023","journal-title":"Neurosci. Biobehav. Rev."},{"key":"ref3","doi-asserted-by":"publisher","first-page":"708","DOI":"10.1097\/WCO.0b013e32835a3432","article-title":"Where, when, and in what form does sporadic Alzheimer's disease begin?","volume":"25","author":"Braak","year":"2012","journal-title":"Curr. Opin. Neurol."},{"key":"ref4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/978-3-319-12679-1_1","article-title":"Neuroanatomy and pathology of sporadic Alzheimer's disease","volume":"215","author":"Braak","year":"2015","journal-title":"Adv. Anat. Embryol. Cell Biol."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"1183","DOI":"10.1038\/s41593-020-0687-6","article-title":"Synergy between amyloid-beta and tau in Alzheimer's disease","volume":"23","author":"Busche","year":"2020","journal-title":"Nat. Neurosci."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"75","DOI":"10.3389\/fnagi.2016.00075","article-title":"Green-fluorescent protein+ astrocytes attach to beta-amyloid plaques in an Alzheimer mouse model and GFP+ astrocytes are sensitive for clasmatodendrosis","volume":"8","author":"Daschil","year":"2016","journal-title":"Front. Aging Neurosci."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"116","DOI":"10.1159\/000285520","article-title":"The vascular hypothesis of Alzheimer's disease: bench to bedside and beyond","volume":"7","author":"de la Torre","year":"2010","journal-title":"Neurodegener Dis"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"35","DOI":"10.3233\/JAD-180004","article-title":"The vascular hypothesis of Alzheimer's disease: a key to preclinical prediction of dementia using neuroimaging","volume":"63","author":"de la Torre","year":"2018","journal-title":"J Alzheimer's Dis"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"413","DOI":"10.1016\/s0079-6123(08)62425-5","article-title":"Role of forebrain cholinergic systems in learning and memory: Relevance to the cognitive deficits of aging and Alzheimer's dementia","volume":"98","author":"Dunnett","year":"1993","journal-title":"Prog Brain Res"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"113","DOI":"10.3389\/fnagi.2018.00113","article-title":"Differential hyperphosphorylation of tau-S199, \u2212T231 and -S396 in organotypic brain slices of Alzheimer mice. A model to study early tau hyperphosphorylation using Okadaic acid","volume":"10","author":"Foidl","year":"2018","journal-title":"Front. Aging Neurosci."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"401","DOI":"10.4103\/1673-5374.266046","article-title":"Can mouse models mimic sporadic Alzheimer\u2019s disease?","volume":"15","author":"Foidl","year":"","journal-title":"Neural Regen. Res."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1016\/j.bbi.2019.01.009","article-title":"Chronic treatment with five vascular risk factors causes cerebral amyloid angiopathy but no Alzheimer pathology in C57BL6 mice","volume":"78","author":"Foidl","year":"","journal-title":"Brain Behav. Immun."},{"key":"ref14","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1016\/0165-0270(81)90003-0","article-title":"Organotypic monolayer cultures of nervous tissue","volume":"4","author":"G\u00e4hwiler","year":"1981","journal-title":"J. Neurosci. Methods"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1007\/978-981-32-9358-8_16","article-title":"Amyloid-beta and tau at the crossroads of Alzheimer's disease","volume":"1184","author":"Gallardo","year":"2019","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"343","DOI":"10.1016\/j.jalz.2011.05.2408","article-title":"Some evolutionary perspectives on Alzheimer's disease pathogenesis and pathology","volume":"8","author":"Glass","year":"2012","journal-title":"Alzheimers Dement."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"fcae369","DOI":"10.1093\/braincomms\/fcae369","article-title":"Cerebral amyloid angiopathy impacts neurofibrillary tangle burden and cognition","volume":"6","author":"Godrich","year":"2024","journal-title":"Brain Commun."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"30","DOI":"10.1038\/s41582-019-0281-2","article-title":"Cerebral amyloid angiopathy and Alzheimer disease - one peptide, two pathways","volume":"16","author":"Greenberg","year":"2020","journal-title":"Nat. Rev. Neurol."},{"key":"ref19","doi-asserted-by":"publisher","first-page":"184","DOI":"10.1126\/science.1566067","article-title":"Alzheimer's disease: the amyloid cascade hypothesis","volume":"256","author":"Hardy","year":"1992","journal-title":"Science"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"908","DOI":"10.2174\/1570159X18666201111104509","article-title":"Glia-driven Neuroinflammation and systemic inflammation in Alzheimer's disease","volume":"19","author":"Hashioka","year":"2021","journal-title":"Curr. Neuropharmacol."},{"key":"ref21","first-page":"309","article-title":"Nerve growth factor is a neurotrophic factor for forebrain cholinergic neurons; implications for Alzheimer's disease","volume":"24","author":"Hefti","year":"1987","journal-title":"J. Neural Transm. Suppl."},{"key":"ref22","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1016\/j.exger.2010.11.032","article-title":"Chronic mild cerebrovascular dysfunction as a cause for Alzheimer's disease? Review","volume":"46","author":"Humpel","year":"2011","journal-title":"Exp. Gerontol."},{"key":"ref23","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1016\/j.neuroscience.2015.07.086","article-title":"Organotypic brain slice cultures - review","volume":"305","author":"Humpel","year":"2015","journal-title":"Neuroscience"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"290","DOI":"10.2174\/1567202614666170705150535","article-title":"Platelets: their potential contribution to the generation of beta-amyloid plaques in Alzheimer\u2019s disease","volume":"14","author":"Humpel","year":"2017","journal-title":"Curr. Neurovasc. Res."},{"key":"ref25","doi-asserted-by":"publisher","first-page":"e59","DOI":"10.1002\/cpim.59","article-title":"Organotypic brain slice cultures","volume":"123","author":"Humpel","year":"2018","journal-title":"Curr. Protoc. Immunol."},{"key":"ref26","doi-asserted-by":"publisher","first-page":"172","DOI":"10.2174\/1567205016666181212153138","article-title":"Organotypic brain slices of adult transgenic mice: a tool to study Alzheimer\u2019s disease","volume":"16","author":"Humpel","year":"2019","journal-title":"Curr. Alzheimer Res."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1007\/978-3-030-74046-7_12","article-title":"NGF released from blood cells or collagen hydrogels as a therapeutic target in Alzheimer\u2019s disease?","volume":"1331","author":"Humpel","year":"2021","journal-title":"Adv Exp Med Biol"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"1881","DOI":"10.4103\/1673-5374.335138","article-title":"Intranasal neprilysin rapidly eliminates beta-amyloid plaques, but causes plaque compensations: the explanation why the beta-amyloid cascade may fail?","volume":"17","author":"Humpel","year":"2022","journal-title":"Neural Regeneration Review"},{"key":"ref29","doi-asserted-by":"publisher","first-page":"1540150","DOI":"10.3389\/fncel.2025.1540150","article-title":"Long-term live-cell imaging of GFAP+ astroglia and laminin+ vessels in organotypic mouse brain slices using microcontact printing","volume":"19","author":"Humpel","year":"2025","journal-title":"Front. Cell. Neurosci."},{"key":"ref30","doi-asserted-by":"publisher","first-page":"253","DOI":"10.1007\/978-3-7091-6139-5_23","article-title":"Nerve growth factor and cholinergic CNS neurons studied in organotypic brain slices: implications in Alzheimer\u2019s disease?","volume":"62","author":"Humpel","year":"2002","journal-title":"J Neural Transmission Supplement"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"451","DOI":"10.2119\/2007-00100.Irvine","article-title":"Protein aggregation in the brain: the molecular basis for Alzheimer's and Parkinson's diseases","volume":"14","author":"Irvine","year":"2008","journal-title":"Mol. Med."},{"key":"ref32","doi-asserted-by":"publisher","first-page":"65","DOI":"10.1038\/nrn2967","article-title":"Amyloid-\u03b2 and tau--a toxic pas de deux in Alzheimer's disease","volume":"12","author":"Ittner","year":"2011","journal-title":"Nat. Rev. Neurosci."},{"key":"ref33","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1080\/19768354.2018.1508498","article-title":"Exploring glia to better understand Alzheimer's disease","volume":"22","author":"Kim","year":"2018","journal-title":"Anim Cells Syst"},{"key":"ref34","doi-asserted-by":"publisher","first-page":"6676","DOI":"10.3390\/ijms21186676","article-title":"Deleterious alteration of glia in the brain of Alzheimer's disease","volume":"21","author":"Kim","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"ref35","doi-asserted-by":"publisher","first-page":"15483","DOI":"10.1038\/s41598-018-33768-2","article-title":"Platelets isolated from an Alzheimer mouse damage healthy cortical vessels and cause inflammation in an organotypic ex vivo brain slice model","volume":"8","author":"Kniewallner","year":"2018","journal-title":"Sci. Rep."},{"key":"ref36","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1097\/WCO.0000000000001330","article-title":"Cerebral amyloid angiopathy: one single entity?","volume":"38","author":"Koemans","year":"2025","journal-title":"Curr. Opin. Neurol."},{"key":"ref37","doi-asserted-by":"publisher","first-page":"1164","DOI":"10.3390\/biom12091164","article-title":"Spreading of P301S aggregated tau investigated in organotypic mouse brain slice cultures","volume":"12","author":"Korde","year":"2022","journal-title":"Biomol. Ther."},{"key":"ref38","doi-asserted-by":"publisher","first-page":"165","DOI":"10.3390\/biom14020165","article-title":"A combination of heavy metals and intracellular pathway modulators induces Alzheimer disease-like pathologies in organotypic brain slices","volume":"14","author":"Korde","year":"2024","journal-title":"Biomol. Ther."},{"key":"ref39","doi-asserted-by":"publisher","first-page":"499","DOI":"10.1038\/nrn2168","article-title":"Intracellular amyloid-beta in Alzheimer's disease","volume":"8","author":"LaFerla","year":"2007","journal-title":"Nat. Rev. Neurosci."},{"key":"ref40","doi-asserted-by":"publisher","first-page":"207","DOI":"10.1016\/0166-4328(93)90137-f","article-title":"Alzheimer's disease: molecular genetics and transgenic animal models","volume":"57","author":"Lannfelt","year":"1993","journal-title":"Behav. Brain Res."},{"key":"ref41","doi-asserted-by":"publisher","first-page":"16","DOI":"10.1006\/exnr.1993.1168","article-title":"Nerve growth factor pharmacology: application to the treatment of cholinergicneurodegeneration in Alzheimer's disease","volume":"124","author":"Lapchak","year":"1993","journal-title":"Exp. Neurol."},{"key":"ref42","first-page":"54","article-title":"The nun study. How one scientist and 678 sisters are helping unlock the secrets of Alzheimer's","volume":"157","author":"Lemonick","year":"2001","journal-title":"Time"},{"key":"ref43","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1007\/978-1-4614-4989-8_19","article-title":"Considering the vascular hypothesis of Alzheimer's disease: effect of copper associated amyloid on red blood cells","volume":"765","author":"Lucas","year":"2013","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref44","doi-asserted-by":"publisher","first-page":"939","DOI":"10.1038\/sj.mp.4002072","article-title":"Beta-amyloid expression, release and extracellular deposition in aged rat brain slices","volume":"13","author":"Marksteiner","year":"2008","journal-title":"Mol. Psychiatry"},{"key":"ref45","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1016\/j.jneumeth.2018.05.021","article-title":"Free-floating adult human brain-derived slice cultures as a model to study the neuronal impact of Alzheimer's disease-associated a\u03b2 oligomers","volume":"307","author":"Mendes","year":"2018","journal-title":"J. Neurosci. Methods"},{"key":"ref46","doi-asserted-by":"publisher","first-page":"606","DOI":"10.1017\/s1092852900016886","article-title":"Metabolic syndrome and Alzheimer's disease: a link to a vascular hypothesis?","volume":"13","author":"Milionis","year":"2008","journal-title":"CNS Spectr."},{"key":"ref47","doi-asserted-by":"publisher","first-page":"434","DOI":"10.3390\/biom11030434","article-title":"Spreading of beta-amyloid in organotypic mouse brain slices and microglial protection of cholinergic neurons","volume":"11","author":"Moelgg","year":"2021","journal-title":"Biomol. Ther."},{"key":"ref48","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1046\/j.1460-9568.2003.02728.x","article-title":"Brain capillaries and cholinergic neurons persist in organotypic brain slices in the absence of blood flow","volume":"18","author":"Moser","year":"2003","journal-title":"Eur. J. Neurosci."},{"key":"ref49","first-page":"80","article-title":"Role of activated glia and of glial cytokines in Alzheimer's disease: a review","volume":"16","author":"Mrak","year":"1996","journal-title":"Eos"},{"key":"ref50","doi-asserted-by":"publisher","first-page":"227","DOI":"10.3109\/00207454.2013.831852","article-title":"Neuroinflammation in Alzheimer's disease: an understanding of physiology and pathology","volume":"124","author":"Obulesu","year":"2014","journal-title":"Int. J. Neurosci."},{"key":"ref51","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1007\/BF02816103","article-title":"Microvascular pathology and vascular basement membrane components in Alzheimer's disease","volume":"9","author":"Perlmutter","year":"1994","journal-title":"Mol. Neurobiol."},{"key":"ref52","doi-asserted-by":"publisher","first-page":"2527","DOI":"10.1007\/s10571-021-01147-7","article-title":"Metabolic regulation of glia and their Neuroinflammatory role in Alzheimer's disease","volume":"42","author":"Preeti","year":"2022","journal-title":"Cell. Mol. Neurobiol."},{"key":"ref53","doi-asserted-by":"publisher","first-page":"461","DOI":"10.1146\/annurev.genet.32.1.461","article-title":"Alzheimer's disease: genetic studies and transgenic models","volume":"32","author":"Price","year":"1998","journal-title":"Annu. Rev. Genet."},{"key":"ref54","doi-asserted-by":"publisher","first-page":"1265","DOI":"10.1161\/ATVBAHA.120.311911","article-title":"Vascular hypothesis of Alzheimer disease: topical review of mouse models","volume":"41","author":"Scheffer","year":"2021","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref55","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1016\/s0047-6374(01)00343-8","article-title":"Glia-related pathomechanisms in Alzheimer's disease: a therapeutic target?","volume":"123","author":"Schubert","year":"2001","journal-title":"Mech. Ageing Dev."},{"key":"ref56","doi-asserted-by":"publisher","first-page":"109979","DOI":"10.1016\/j.jneumeth.2023.109979","article-title":"Long-term organotypic brain slices cultured on collagen-based microcontact prints: a perspective for a brain-on-a-chip","volume":"399","author":"Steiner","year":"2023","journal-title":"J. Neurosci. Methods"},{"key":"ref57","doi-asserted-by":"publisher","first-page":"1109","DOI":"10.3390\/biom14091109","article-title":"From Organotypic mouse brain slices to human Alzheimer plasma biomarkers: a focus on microglia","volume":"14","author":"Steiner","year":"2024","journal-title":"Biomol. Ther."},{"key":"ref58","doi-asserted-by":"publisher","first-page":"173","DOI":"10.1016\/0165-0270(91)90128-m","article-title":"A simple method for organotypic cultures of nervous tissue","volume":"37","author":"Stoppini","year":"1991","journal-title":"J. Neurosci. Methods"},{"key":"ref59","doi-asserted-by":"publisher","first-page":"182","DOI":"10.1007\/s12640-011-9272-9","article-title":"Alzheimer's disease pathologic cascades: who comes first, what drives what","volume":"22","author":"Swerdlow","year":"2012","journal-title":"Neurotox. Res."},{"key":"ref60","doi-asserted-by":"publisher","first-page":"595","DOI":"10.4103\/1673-5374.230273","article-title":"Collagen for brain repair: therapeutic perspectives - a mini-review","volume":"13","author":"Ucar","year":"2018","journal-title":"Neural Regen. Res."},{"key":"ref61","doi-asserted-by":"publisher","first-page":"163","DOI":"10.3390\/biom12020163","article-title":"Spreading of aggregated \u03b1 synuclein in sagittal organotypic mouse brain slices","volume":"12","author":"Ucar","year":"2022","journal-title":"Biomol. Ther."},{"key":"ref62","doi-asserted-by":"publisher","first-page":"2521","DOI":"10.1007\/s00221-020-05907-7","article-title":"Collagen hydrogels loaded with fibroblast growth factor-2 as a bridge to repair brain vessels in organotypic brain slices","volume":"238","author":"Ucar","year":"2020","journal-title":"Exp. Brain Res."},{"key":"ref63","doi-asserted-by":"publisher","first-page":"105466","DOI":"10.1016\/j.ebiom.2024.105466","article-title":"The role of neuroinflammation in cerebral amyloid angiopathy","volume":"110","author":"van den Brink","year":"2024","journal-title":"EBioMedicine"},{"key":"ref64","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1161\/01.str.18.2.311","article-title":"Cerebral amyloid angiopathy. A critical review","volume":"18","author":"Vinters","year":"1987","journal-title":"Stroke"},{"key":"ref65","doi-asserted-by":"publisher","first-page":"303","DOI":"10.1016\/B978-0-444-63945-5.00016-7","article-title":"Prion-like mechanisms in Alzheimer disease","volume":"153","author":"Walker","year":"2018","journal-title":"Handb. Clin. Neurol."},{"key":"ref66","doi-asserted-by":"publisher","first-page":"251","DOI":"10.1038\/nrn.2016.13","article-title":"A critical appraisal of the pathogenic protein spread hypothesis of neurodegeneration.Nat","volume":"17","author":"Walsh","year":"2016","journal-title":"Rev. Neurosci."},{"key":"ref67","doi-asserted-by":"publisher","first-page":"787","DOI":"10.1038\/s41422-019-0216-x","article-title":"Sodium oligomannate therapeutically remodels gut microbiota and suppresses gut bacterial amino acids-shaped neuroinflammation to inhibit Alzheimer\u2019s disease progression","volume":"29","author":"Wang","year":"2019","journal-title":"Cell Res."},{"key":"ref68","doi-asserted-by":"publisher","DOI":"10.31083\/FBL36257","article-title":"Novel Plasma Biomarkers for Alzheimer\u2019s Disease: Insights from Organotypic Brain Slice and Microcontact Printing Techniques","author":"Yilmaz","year":"2025","journal-title":"Front. Biosci. (Landmark Ed)."},{"key":"ref69","doi-asserted-by":"publisher","first-page":"1326","DOI":"10.3390\/biom14101326","article-title":"From Organotypic mouse brain slices to human Alzheimer\u2019s plasma biomarkers: a focus on nerve Fiber outgrowth","volume":"14","author":"Yilmaz","year":"2024","journal-title":"Biomol. Ther."}],"container-title":["Frontiers in Dementia"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/frdem.2025.1585124\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T06:13:10Z","timestamp":1753423990000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/frdem.2025.1585124\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,25]]},"references-count":68,"alternative-id":["10.3389\/frdem.2025.1585124"],"URL":"https:\/\/doi.org\/10.3389\/frdem.2025.1585124","relation":{},"ISSN":["2813-3919"],"issn-type":[{"value":"2813-3919","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,25]]},"article-number":"1585124"}}