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pmid:34868234
Identification of a Qualitative Signature for the Diagnosis of Dementia With Lewy Bodies.
Shu Zhou · Qingchun Meng · Lingyu Li · Luo Hai · Zexuan Wang · Zhicheng Li · Yingli Sun
Frontiers in genetics · 2021 doi:10.3389/fgene.2021.758103
Paper
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Experiment
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Data
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Analysis
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Execution
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Complete file
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We aimed to develop a blood-based qualitative signature for differentiating DLB patients from healthy controls. Methods: The GSE120584 dataset was downloaded from the public database Gene Expression Omnibus (GEO). We combined multiple methods to select features based on the within-sample relative expression orderings (REOs) of microRNA (miRNA) pairs. Specifically, we first quickly selected miRNA pairs related to DLB by identifying reversal stable miRNA pairs. Then, an optimal miRNA pair subset was extracted by random forest (RF) and support vector machine-recursive feature elimination (SVM-RFE) methods. Furthermore, we applied logistic regression (LR) and SVM to build several prediction models. The model performance was assessed using the receiver operating characteristic curve (ROC) analysis. Lastly, we conducted bioinformatics analyses to explore the molecular mechanisms of the discovered miRNAs. Results: A qualitative signature consisted of 17 miRNA pairs and two clinical factors was identified for discriminating DLB patients from healthy controls. The signature is robust against experimental batch effects and applicable at the individual levels. The accuracies of the-signature-based models on the test set are 82.61 and 79.35%, respectively, indicating that the signature has acceptable discrimination performance. Moreover, bioinformatics analyses revealed that predicted target genes were enriched in 11 Go terms and 2 KEGG pathways. Moreover, five potential hub genes were found for DLB, including SRF, MAPK1, YWHAE, RPS6KA3, and KDM7A. Conclusion: This study provided a blood-based qualitative signature with the potential to be used as an effective tool to improve the accuracy of DLB diagnosis.", "claims": [{"id": "claim-method", "statement": "The model performance was assessed using the receiver operating characteristic curve (ROC) analysis.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:34868234", "locator": "sections[0] \"Abstract\" paragraph 2", "quote": "The model performance was assessed using the receiver operating characteristic curve (ROC) analysis."}], "state": "complete"}, {"id": "claim-data-access", "statement": "Finally, only one dataset GSE120584 was screened out and downloaded.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:34868234", "locator": "sections[3] \"Dataset Collection and Preprocessing\" paragraph 2", "quote": "Finally, only one dataset GSE120584 was screened out and downloaded."}], "state": "complete"}, {"id": "claim-target-result", "statement": "The accuracies of the-signature-based models on the test set are 82.61 and 79.35%, respectively, indicating that the signature has acceptable discrimination performance.", "kind": "quantitative", "target": {"value": 79.35, "unit": "per cent", "tolerance": null, "source": "The accuracies of the-signature-based models on the test set are 82.61 and 79.35%, respectively, indicating that the signature has acceptable discrimination performance."}, "evidence": [{"asset": "source:pmid:34868234", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "The accuracies of the-signature-based models on the test set are 82.61 and 79.35%, respectively, indicating that the signature has acceptable discrimination performance."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "partial", "objective": "The accuracies of the-signature-based models on the test set are 82.61 and 79.35%, respectively, indicating that the signature has acceptable discrimination performance.", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:34868234"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. This lane did not download the dataset or read its licence.", "duration_hours": null, "parameters": {}, "inputs": ["data:geo:GSE120584"], "outputs": ["the source data with its repository provenance"], "instruments": [], "reagents": []}, {"id": "run_analysis", "order": 3, "name": "Run the published analysis", "kind": "analysis", "state": "blocked", "blocked_on": "method:machine_learning_random_forest", "description": "Apply the paper's statistical method to the resolved series and compare the result with the published target.", "duration_hours": null, "parameters": {}, "inputs": ["data:geo:GSE120584"], "outputs": ["the replicated result and its comparison"], "instruments": [], "reagents": []}], "materials": [], "instruments": [], "reagents": [], "conditions": {}, "gaps": ["step:run_analysis blocked on method:machine_learning_random_forest"]}, "data": {"state": "partial", "assets": [{"id": "source:pmid:34868234", "role": "manuscript", "format": "xml", "media_type": "application/xml", 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"GSM3404452", "GSM3404453", "GSM3404454", "GSM3404455", "GSM3404456", "GSM3404457", "GSM3404458", "GSM3404459", "GSM3404460", "GSM3404461", "GSM3404462", "GSM3404463", "GSM3404464", "GSM3404465", "GSM3404466", "GSM3404467", "GSM3404468", "GSM3404469", "GSM3404470", "GSM3404471", "GSM3404472", "GSM3404473", "GSM3404474", "GSM3404475", "GSM3404476", "GSM3404477", "GSM3404478", "GSM3404479", "GSM3404480", "GSM3404481", "GSM3404482", "GSM3404483", "GSM3404484", "GSM3404485", "GSM3404486", "GSM3404487", "GSM3404488", "GSM3404489", "GSM3404490", "GSM3404491", "GSM3404492", "GSM3404493", "GSM3404494", "GSM3404495", "GSM3404496", "GSM3404497", "GSM3404498", "GSM3404499", "GSM3404500", "GSM3404501", "GSM3404502", "GSM3404503", "GSM3404504", "GSM3404505", "GSM3404506", "GSM3404507", "GSM3404508", "GSM3404509", "GSM3404510", "GSM3404511", "GSM3404512", "GSM3404513", "GSM3404514", "GSM3404515", "GSM3404516", "GSM3404517", "GSM3404518", "GSM3404519", "GSM3404520", "GSM3404521", "GSM3404522", "GSM3404523", "GSM3404524", "GSM3404525", "GSM3404526", "GSM3404527", "GSM3404528", "GSM3404529", "GSM3404530", "GSM3404531", "GSM3404532", "GSM3404533", "GSM3404534", "GSM3404535", "GSM3404536", "GSM3404537", "GSM3404538", "GSM3404539", "GSM3404540", "GSM3404541", "GSM3404542", "GSM3404543", "GSM3404544", "GSM3404545", "GSM3404546", "GSM3404547", "GSM3404548", "GSM3404549", "GSM3404550", "GSM3404551", "GSM3404552", "GSM3404553", "GSM3404554", "GSM3404555", "GSM3404556", "GSM3404557", "GSM3404558", "GSM3404559", "GSM3404560", "GSM3404561", "GSM3404562", "GSM3404563", "GSM3404564", "GSM3404565", "GSM3404566", "GSM3404567", "GSM3404568", "GSM3404569", "GSM3404570", "GSM3404571", "GSM3404572", "GSM3404573", "GSM3404574", "GSM3404575", "GSM3404576", "GSM3404577", "GSM3404578", "GSM3404579", "GSM3404580", "GSM3404581", "GSM3404582", "GSM3404583", "GSM3404584", "GSM3404585", "GSM3404586", "GSM3404587", "GSM3404588", "GSM3404589", "GSM3404590", "GSM3404591", "GSM3404592", "GSM3404593", "GSM3404594", "GSM3404595", "GSM3404596", "GSM3404597", "GSM3404598", "GSM3404599", "GSM3404600", "GSM3404601", "GSM3404602", "GSM3404603", "GSM3404604", "GSM3404605", "GSM3404606", "GSM3404607", "GSM3404608", "GSM3404609", "GSM3404610", "GSM3404611", "GSM3404612", "GSM3404613", "GSM3404614", "GSM3404615", "GSM3404616", "GSM3404617", "GSM3404618", "GSM3404619", "GSM3404620", "GSM3404621", "GSM3404622", "GSM3404623", "GSM3404624", "GSM3404625", "GSM3404626", "GSM3404627", "GSM3404628", "GSM3404629", "GSM3404630", "GSM3404631", "GSM3404632", "GSM3404633", "GSM3404634", "GSM3404635", "GSM3404636", "GSM3404637", "GSM3404638", "GSM3404639", "GSM3404640", "GSM3404641", "GSM3404642", "GSM3404643", "GSM3404644", "GSM3404645", "GSM3404646", "GSM3404647", "GSM3404648", "GSM3404649", "GSM3404650", "GSM3404651", "GSM3404652", "GSM3404653", "GSM3404654", "GSM3404655", "GSM3404656", "GSM3404657", "GSM3404658", "GSM3404659", "GSM3404660", "GSM3404661", "GSM3404662", "GSM3404663", "GSM3404664", "GSM3404665", "GSM3404666", "GSM3404667", "GSM3404668", "GSM3404669", "GSM3404670", "GSM3404671", "GSM3404672", "GSM3404673", "GSM3404674", "GSM3404675", "GSM3404676", "GSM3404677", "GSM3404678", "GSM3404679", "GSM3404680", "GSM3404681", "GSM3404682", "GSM3404683", "GSM3404684", "GSM3404685", "GSM3404686", "GSM3404687", "GSM3404688", "GSM3404689", "GSM3404690", "GSM3404691", "GSM3404692", "GSM3404693", "GSM3404694", "GSM3404695", "GSM3404696", "GSM3404697", "GSM3404698", "GSM3404699", "GSM3404700", "GSM3404701", "GSM3404702", "GSM3404703", "GSM3404704", "GSM3404705", "GSM3404706", "GSM3404707", "GSM3404708", "GSM3404709", "GSM3404710", "GSM3404711", "GSM3404712", "GSM3404713", "GSM3404714", "GSM3404715", "GSM3404716", "GSM3404717", "GSM3404718", "GSM3404719", "GSM3404720", "GSM3404721", "GSM3404722", "GSM3404723", "GSM3404724", "GSM3404725", "GSM3404726", "GSM3404727", "GSM3404728", "GSM3404729", "GSM3404730", "GSM3404731", "GSM3404732", "GSM3404733", "GSM3404734", "GSM3404735", "GSM3404736", "GSM3404737", "GSM3404738", "GSM3404739", "GSM3404740", "GSM3404741", "GSM3404742", "GSM3404743", "GSM3404744", "GSM3404745", "GSM3404746", "GSM3404747", "GSM3404748", "GSM3404749", "GSM3404750", "GSM3404751", "GSM3404752", "GSM3404753", "GSM3404754", "GSM3404755", "GSM3404756", "GSM3404757", "GSM3404758", "GSM3404759", "GSM3404760", "GSM3404761", "GSM3404762", "GSM3404763", "GSM3404764", "GSM3404765", "GSM3404766", "GSM3404767", "GSM3404768", "GSM3404769", "GSM3404770", "GSM3404771", "GSM3404772", "GSM3404773", "GSM3404774", "GSM3404775", "GSM3404776", "GSM3404777", "GSM3404778", "GSM3404779", "GSM3404780", "GSM3404781", "GSM3404782", "GSM3404783", "GSM3404784", "GSM3404785", "GSM3404786", "GSM3404787", "GSM3404788", "GSM3404789", "GSM3404790", "GSM3404791", "GSM3404792", "GSM3404793", "GSM3404794", "GSM3404795", "GSM3404796", "GSM3404797", "GSM3404798", "GSM3404799", "GSM3404800", "GSM3404801", "GSM3404802", "GSM3404803", "GSM3404804", "GSM3404805", "GSM3404806", "GSM3404807", "GSM3404808", "GSM3404809", "GSM3404810", "GSM3404811", "GSM3404812", "GSM3404813", "GSM3404814", "GSM3404815", "GSM3404816", "GSM3404817", "GSM3404818", "GSM3404819", "GSM3404820", "GSM3404821", "GSM3404822", "GSM3404823", "GSM3404824", "GSM3404825", "GSM3404826", "GSM3404827", "GSM3404828", "GSM3404829", "GSM3404830", "GSM3404831", "GSM3404832", "GSM3404833", "GSM3404834", "GSM3404835", "GSM3404836", "GSM3404837", "GSM3404838", "GSM3404839", "GSM3404840", "GSM3404841", "GSM3404842", "GSM3404843", "GSM3404844", "GSM3404845", "GSM3404846", "GSM3404847", "GSM3404848", "GSM3404849", "GSM3404850", "GSM3404851", "GSM3404852", "GSM3404853", "GSM3404854", "GSM3404855", "GSM3404856", "GSM3404857", "GSM3404858", "GSM3404859", "GSM3404860", "GSM3404861", "GSM3404862", "GSM3404863", "GSM3404864", "GSM3404865", "GSM3404866", "GSM3404867", "GSM3404868", "GSM3404869", "GSM3404870", "GSM3404871", "GSM3404872", "GSM3404873", "GSM3404874", "GSM3404875", "GSM3404876", "GSM3404877", "GSM3404878", "GSM3404879", "GSM3404880", "GSM3404881", "GSM3404882", "GSM3404883", "GSM3404884", "GSM3404885", "GSM3404886", "GSM3404887", "GSM3404888", "GSM3404889", "GSM3404890", "GSM3404891", "GSM3404892", "GSM3404893", "GSM3404894", "GSM3404895", "GSM3404896", "GSM3404897", "GSM3404898", "GSM3404899", "GSM3404900", "GSM3404901", "GSM3404902", "GSM3404903", "GSM3404904", "GSM3404905", "GSM3404906", "GSM3404907", "GSM3404908", "GSM3404909", "GSM3404910", "GSM3404911", "GSM3404912", "GSM3404913", "GSM3404914", "GSM3404915", "GSM3404916", "GSM3404917", "GSM3404918", "GSM3404919", "GSM3404920", "GSM3404921", "GSM3404922", "GSM3404923", "GSM3404924", "GSM3404925", "GSM3404926", "GSM3404927", "GSM3404928", "GSM3404929", "GSM3404930", "GSM3404931", "GSM3404932", "GSM3404933", "GSM3404934", "GSM3404935", "GSM3404936", "GSM3404937", "GSM3404938", "GSM3404939", "GSM3404940", "GSM3404941", "GSM3404942", "GSM3404943", "GSM3404944", "GSM3404945", "GSM3404946", "GSM3404947", "GSM3404948", "GSM3404949", "GSM3404950", "GSM3404951", "GSM3404952", "GSM3404953", "GSM3404954", "GSM3404955", "GSM3404956", "GSM3404957", "GSM3404958", "GSM3404959", "GSM3404960", "GSM3404961", "GSM3404962", "GSM3404963", "GSM3404964", "GSM3404965", "GSM3404966", "GSM3404967", "GSM3404968", "GSM3404969", "GSM3404970", "GSM3404971", "GSM3404972", "GSM3404973", "GSM3404974", "GSM3404975", "GSM3404976", "GSM3404977", "GSM3404978", "GSM3404979", "GSM3404980", "GSM3404981", "GSM3404982", "GSM3404983", "GSM3404984", "GSM3404985", "GSM3404986", "GSM3404987", "GSM3404988", "GSM3404989", "GSM3404990", "GSM3404991", "GSM3404992", "GSM3404993", "GSM3404994", "GSM3404995", "GSM3404996", "GSM3404997", "GSM3404998", "GSM3404999", "GSM3405000", "GSM3405001", "GSM3405002", "GSM3405003", "GSM3405004", "GSM3405005", "GSM3405006", "GSM3405007", "GSM3405008", "GSM3405009", "GSM3405010", "GSM3405011", "GSM3405012", "GSM3405013", "GSM3405014", "GSM3405015", "GSM3405016", "GSM3405017", "GSM3405018", "GSM3405019", "GSM3405020", "GSM3405021", "GSM3405022", "GSM3405023", "GSM3405024", "GSM3405025", "GSM3405026", "GSM3405027", "GSM3405028", "GSM3405029", "GSM3405030", "GSM3405031", "GSM3405032", "GSM3405033", "GSM3405034", "GSM3405035", "GSM3405036", "GSM3405037", "GSM3405038", "GSM3405039", "GSM3405040", "GSM3405041", "GSM3405042", "GSM3405043", "GSM3405044", "GSM3405045", "GSM3405046", "GSM3405047", "GSM3405048", "GSM3405049", "GSM3405050", "GSM3405051", "GSM3405052", "GSM3405053", "GSM3405054", "GSM3405055", "GSM3405056", "GSM3405057", "GSM3405058", "GSM3405059", "GSM3405060", "GSM3405061", "GSM3405062", "GSM3405063", "GSM3405064", "GSM3405065", "GSM3405066", "GSM3405067", "GSM3405068", "GSM3405069", "GSM3405070", "GSM3405071", "GSM3405072", "GSM3405073", "GSM3405074", "GSM3405075", "GSM3405076", "GSM3405077", "GSM3405078", "GSM3405079", "GSM3405080", "GSM3405081", "GSM3405082", "GSM3405083", "GSM3405084", "GSM3405085", "GSM3405086", "GSM3405087", "GSM3405088", "GSM3405089", "GSM3405090", "GSM3405091", "GSM3405092", "GSM3405093", "GSM3405094", "GSM3405095", "GSM3405096", "GSM3405097", "GSM3405098", "GSM3405099", "GSM3405100", "GSM3405101", "GSM3405102", "GSM3405103", "GSM3405104", "GSM3405105", "GSM3405106", "GSM3405107", "GSM3405108", "GSM3405109", "GSM3405110", "GSM3405111", "GSM3405112", "GSM3405113", "GSM3405114", "GSM3405115", "GSM3405116", "GSM3405117", "GSM3405118", "GSM3405119", "GSM3405120", "GSM3405121", "GSM3405122", "GSM3405123", "GSM3405124", "GSM3405125", "GSM3405126", "GSM3405127", "GSM3405128", "GSM3405129", "GSM3405130", "GSM3405131", "GSM3405132", "GSM3405133", "GSM3405134", "GSM3405135", "GSM3405136", "GSM3405137", "GSM3405138", "GSM3405139", "GSM3405140", "GSM3405141", "GSM3405142", "GSM3405143", "GSM3405144", "GSM3405145", "GSM3405146", "GSM3405147", "GSM3405148", "GSM3405149", "GSM3405150", "GSM3405151", "GSM3405152", "GSM3405153", "GSM3405154", "GSM3405155", "GSM3405156", "GSM3405157", "GSM3405158", "GSM3405159", "GSM3405160", "GSM3405161", "GSM3405162", "GSM3405163", "GSM3405164", "GSM3405165", "GSM3405166", "GSM3405167", "GSM3405168", "GSM3405169", "GSM3405170", "GSM3405171", "GSM3405172", "GSM3405173", "GSM3405174", "GSM3405175", "GSM3405176", "GSM3405177", "GSM3405178", "GSM3405179", "GSM3405180", "GSM3405181", "GSM3405182", "GSM3405183", "GSM3405184", "GSM3405185", "GSM3405186", "GSM3405187", "GSM3405188", "GSM3405189", "GSM3405190", "GSM3405191", "GSM3405192", "GSM3405193", "GSM3405194", "GSM3405195", "GSM3405196", "GSM3405197", "GSM3405198", "GSM3405199", "GSM3405200", "GSM3405201", "GSM3405202", "GSM3405203", "GSM3405204", "GSM3405205", "GSM3405206", "GSM3405207", "GSM3405208", "GSM3405209", "GSM3405210", "GSM3405211", "GSM3405212", "GSM3405213", "GSM3405214", "GSM3405215", "GSM3405216", "GSM3405217", "GSM3405218", "GSM3405219", "GSM3405220", "GSM3405221", "GSM3405222", "GSM3405223", "GSM3405224", "GSM3405225", "GSM3405226", "GSM3405227", "GSM3405228", "GSM3405229", "GSM3405230", "GSM3405231", "GSM3405232", "GSM3405233", "GSM3405234", "GSM3405235", "GSM3405236", "GSM3405237", "GSM3405238", "GSM3405239", "GSM3405240", "GSM3405241", "GSM3405242", "GSM3405243", "GSM3405244", "GSM3405245", "GSM3405246", "GSM3405247", "GSM3405248", "GSM3405249", "GSM3405250", "GSM3405251", "GSM3405252", "GSM3405253", "GSM3405254", "GSM3405255", "GSM3405256", "GSM3405257", "GSM3405258", "GSM3405259", "GSM3405260", "GSM3405261", "GSM3405262", "GSM3405263", "GSM3405264", "GSM3405265", "GSM3405266", "GSM3405267", "GSM3405268", "GSM3405269", "GSM3405270", "GSM3405271", "GSM3405272", "GSM3405273", "GSM3405274", "GSM3405275", "GSM3405276", "GSM3405277", "GSM3405278", "GSM3405279", "GSM3405280", "GSM3405281", "GSM3405282", "GSM3405283", "GSM3405284", "GSM3405285", "GSM3405286", "GSM3405287", "GSM3405288", "GSM3405289", "GSM3405290", "GSM3405291", "GSM3405292", "GSM3405293", "GSM3405294", "GSM3405295", "GSM3405296", "GSM3405297", "GSM3405298", "GSM3405299", "GSM3405300", "GSM3405301", "GSM3405302", "GSM3405303", "GSM3405304", "GSM3405305", "GSM3405306", "GSM3405307", "GSM3405308", "GSM3405309", "GSM3405310", "GSM3405311", "GSM3405312", "GSM3405313", "GSM3405314", "GSM3405315", "GSM3405316", "GSM3405317", "GSM3405318", "GSM3405319", "GSM3405320", "GSM3405321", "GSM3405322", "GSM3405323", "GSM3405324", "GSM3405325", "GSM3405326", "GSM3405327", "GSM3405328", "GSM3405329", "GSM3405330", "GSM3405331", "GSM3405332", "GSM3405333", "GSM3405334", "GSM3405335", "GSM3405336", "GSM3405337", "GSM3405338", "GSM3405339", "GSM3405340", "GSM3405341", "GSM3405342", "GSM3405343", "GSM3405344", "GSM3405345", "GSM3405346", "GSM3405347", "GSM3405348", "GSM3405349", "GSM3405350", "GSM3405351", "GSM3405352", "GSM3405353", "GSM3405354", "GSM3405355", "GSM3405356", "GSM3405357", "GSM3405358", "GSM3405359", "GSM3405360", "GSM3405361"], "sample_count": 1601, "group_sizes": null, "group_sizes_basis": null, "platform": "21263", "taxon": "Homo sapiens", "title": "Risk prediction models for dementia constructed by supervised principal component analysis using miRNA expression data", "resolution_checked_at": "2026-11-01T17:30:42Z", "resolution_check": "identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}], "gaps": ["asset:data:geo:GSE120584 not downloaded", "asset:data:geo:GSE120584 licence not read"]}, "analysis": {"state": "partial", "methods": [{"id": "method-1", "family": "machine_learning", "specific": "random_forest", "label": "Machine learning", "dsl_operations": [], "missing_operations": ["machine_learning_random_forest"], "expressible": false, "parameters": {}, "data_shapes": ["labelled_feature_matrix"], "evidence": {"score": 0.7, "band": "medium", "quote": "random forest"}}, {"id": "method-2", "family": "machine_learning", "specific": "support_vector_machine", "label": "Machine learning", "dsl_operations": ["train_classifier", "cross_validate", "roc_auc"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["labelled_feature_matrix"], "evidence": {"score": 0.7, "band": "medium", "quote": "support vector machine"}}, {"id": "method-3", "family": "regression_modelling", "specific": "logistic", "label": "Regression modelling", "dsl_operations": ["glm_logit_fit"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["predictors_and_outcome"], "evidence": {"score": 0.7, "band": "medium", "quote": "logistic regression"}}, {"id": "method-4", "family": null, "specific": null, "label": "receiver operating characteristic curve (ROC) analysis", "dsl_operations": [], "missing_operations": [], "expressible": null, "parameters": {}, "data_shapes": [], "evidence": {"score": 1.0, "band": "verbatim", "quote": "The model performance was assessed using the receiver operating characteristic curve (ROC) analysis."}}], "targets": [{"id": "target-published-estimate", "claim_id": "claim-target-result", "operation": "cross_validate", "expected_value": 79.35, "unit": "per cent", "tolerance": null, "state": "complete", "note": "The published estimate as the reader quoted it from the paper. 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