{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,20]],"date-time":"2025-11-20T12:13:58Z","timestamp":1763640838142,"version":"3.45.0"},"reference-count":51,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2025,11,20]],"date-time":"2025-11-20T00:00:00Z","timestamp":1763596800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Saudi Ministry of Higher Education and Prince Sattam bin Abdulaziz University for Fatimah Qassadi","award":["A92696"],"award-info":[{"award-number":["A92696"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Recurrent Clostridioides difficile infection (rCDI) remains a major therapeutic challenge. Although faecal microbiota transplantation (FMT) is highly effective and thought to restore microbial composition and metabolic function, the mechanisms underlying its success are not fully understood. In particular, the contribution of non-bacterial components such as soluble metabolites remains unclear. Therefore, further investigation is needed to identify the mechanistic drivers of FMT efficacy and clarify how non-bacterial factors contribute to therapeutic outcomes. Here, we applied untargeted three-dimensional Orbitrap secondary ion mass spectrometry (3D OrbiSIMS) to profile faecal metabolic reprogramming in rCDI patients pre- and post-FMT, alongside C. difficile cultures exposed to sterile faecal filtrates. FMT induced extensive metabolic shifts, restoring glyoxylate\/dicarboxylate and glycerophosphoinositol pathways and normalising disrupted bile acid and amino acid profiles. Faecal filtrate exposure caused strain-specific metabolic disruption in C. difficile, depleting proline, fumarate and succinate while enriching tryptophan. While multiple metabolite classes were profiled, the most significant functional changes were observed in lipids. Lipidomics identified &gt;3.8-fold enrichment of phosphatidylinositol (PI) species, which localised to bacterial membranes and conferred cytoprotection against C. difficile toxins and other epithelial insults. Spatial metabolomics imaging revealed, for the first time, metabolite compartmentalisation within C. difficile, with proline and succinate broadly distributed across the cell surface and fumarate confined to distinct microdomains, highlighting functional heterogeneity in pathogen metabolism. Collectively, these findings demonstrate that soluble metabolites within faecal filtrates mediate pathogen suppression and epithelial barrier protection, establishing metabolite-driven mechanisms underlying FMT efficacy and identifying PI lipids as candidate post-biotic therapeutics for rCDI.<\/jats:p>","DOI":"10.3390\/ijms262211221","type":"journal-article","created":{"date-parts":[[2025,11,20]],"date-time":"2025-11-20T12:02:42Z","timestamp":1763640162000},"page":"11221","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Untargeted Metabolomics Identifies Faecal Filtrate-Derived Metabolites That Disrupt Clostridioides difficile Metabolism and Confer Gut Barrier Cytoprotection"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9260-2882","authenticated-orcid":false,"given":"Fatimah I.","family":"Qassadi","sequence":"first","affiliation":[{"name":"School of Pharmacy, Prince Sattam bin Abdulaziz University, Al-Kharj 11942, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Charlotte","family":"Johnson","sequence":"additional","affiliation":[{"name":"Nottingham Digestive Diseases Centre, Translational Medical Sciences Unit, School of Medicine, University of Nottingham, Nottingham NG7 2UH, UK"},{"name":"Biodiscovery Institute, School of Medicine, University of Nottingham, Nottingham NG7 2RD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6329-9340","authenticated-orcid":false,"given":"Karen","family":"Robinson","sequence":"additional","affiliation":[{"name":"Nottingham Digestive Diseases Centre, Translational Medical Sciences Unit, School of Medicine, University of Nottingham, Nottingham NG7 2UH, UK"},{"name":"Biodiscovery Institute, School of Medicine, University of Nottingham, Nottingham NG7 2RD, UK"},{"name":"NIHR Nottingham Biomedical Research Centre, University of Nottingham, Nottingham NG7 2UH, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0269-059X","authenticated-orcid":false,"given":"Ruth","family":"Griffin","sequence":"additional","affiliation":[{"name":"NIHR Nottingham Biomedical Research Centre, University of Nottingham, Nottingham NG7 2UH, UK"},{"name":"Vaccines and Therapeutics Group, Biodiscovery Institute, School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christos","family":"Polytarchou","sequence":"additional","affiliation":[{"name":"Department of Biosciences, Centre for Systems Health and Integrated Metabolic Research (SHiMR), Nottingham Trent University, Nottingham NG11 8NS, UK"},{"name":"John van Geest Cancer Research Centre, School of Science & Technology, Nottingham Trent University, Nottingham NG11 8NS, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6096-0586","authenticated-orcid":false,"given":"Dina","family":"Kao","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology, Department of Medicine, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB T6G 2R3, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3689-2130","authenticated-orcid":false,"given":"Dong-Hyun","family":"Kim","sequence":"additional","affiliation":[{"name":"School of Pharmacy, University of Nottingham, Nottingham NG7 2RD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1601-4664","authenticated-orcid":false,"given":"Rian L.","family":"Griffiths","sequence":"additional","affiliation":[{"name":"School of Pharmacy, University of Nottingham, Nottingham NG7 2RD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2135-3812","authenticated-orcid":false,"given":"Zheying","family":"Zhu","sequence":"additional","affiliation":[{"name":"School of Pharmacy, University of Nottingham, Nottingham NG7 2RD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7622-3997","authenticated-orcid":false,"given":"Tanya M.","family":"Monaghan","sequence":"additional","affiliation":[{"name":"Nottingham Digestive Diseases Centre, Translational Medical Sciences Unit, School of Medicine, University of Nottingham, Nottingham NG7 2UH, UK"},{"name":"NIHR Nottingham Biomedical Research Centre, University of Nottingham, Nottingham NG7 2UH, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Feuerstadt, P., Theriault, N., and Tillotson, G. (2023). The burden of CDI in the United States: A multifactorial challenge. BMC Infect. Dis., 23.","DOI":"10.1186\/s12879-023-08096-0"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1016\/S0140-6736(19)31266-8","article-title":"The evolution of the use of faecal microbiota transplantation and emerging therapeutic indications","volume":"394","author":"Allegretti","year":"2019","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"100642","DOI":"10.1016\/j.eclinm.2020.100642","article-title":"Faecal microbiota transplantation for recurrent Clostridioides difficile infection: An updated systematic review and meta-analysis","volume":"29\u201330","author":"Baunwall","year":"2020","journal-title":"eClinicalMedicine"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1016\/j.chom.2023.03.019","article-title":"Beneficial effects of fecal microbiota transplantation in recurrent Clostridioides difficile infection","volume":"31","author":"Yadegar","year":"2023","journal-title":"Cell Host Microbe"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1053\/j.gastro.2016.11.010","article-title":"Efficacy of Sterile Fecal Filtrate Transfer for Treating Patients with Clostridium difficile Infection","volume":"152","author":"Ott","year":"2017","journal-title":"Gastroenterology"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"986","DOI":"10.1016\/S2468-1253(25)00190-6","article-title":"Effects of lyophilised faecal filtrate compared with lyophilised donor stool on Clostridioides difficile recurrence: A multicentre, randomised, double-blinded, non-inferiority trial","volume":"10","author":"Kao","year":"2025","journal-title":"Lancet Gastroenterol. Hepatol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/j.tim.2012.04.001","article-title":"Interaction between the intestinal microbiota and host in Clostridium difficile colonization resistance","volume":"20","author":"Britton","year":"2012","journal-title":"Trends Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"117248","DOI":"10.1016\/j.trac.2023.117248","article-title":"Advances in the integration of metabolomics and metagenomics for human gut microbiome and their clinical applications","volume":"167","author":"Yang","year":"2023","journal-title":"TrAC Trends Anal. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1038\/s41564-019-0664-6","article-title":"Spatial metabolomics of in situ host\u2013microbe interactions at the micrometre scale","volume":"5","author":"Geier","year":"2020","journal-title":"Nat. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.aca.2018.05.031","article-title":"A review on human fecal metabolomics: Methods, applications and the human fecal metabolome database","volume":"1030","author":"Karu","year":"2018","journal-title":"Anal. Chim. Acta"},{"key":"ref_11","first-page":"1","article-title":"Chapter One-Analytical Platforms in Metabolomics of Health and Disease","volume":"Volume 104","author":"Ratnasekhar","year":"2024","journal-title":"Comprehensive Analytical Chemistry"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"999","DOI":"10.1002\/sia.6907","article-title":"Surface-sampling mass spectrometry and imaging: Direct analysis of bacterial species","volume":"53","author":"Griffiths","year":"2021","journal-title":"Surf. Interface Anal."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1038\/nmeth.4504","article-title":"The 3D OrbiSIMS\u2014Label-free metabolic imaging with subcellular lateral resolution and high mass-resolving power","volume":"14","author":"Passarelli","year":"2017","journal-title":"Nat. Methods"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1985","DOI":"10.1001\/jama.2017.17077","article-title":"Effect of Oral Capsule- vs Colonoscopy-Delivered Fecal Microbiota Transplantation on Recurrent Clostridium difficile Infection: A Randomized Clinical Trial","volume":"318","author":"Kao","year":"2017","journal-title":"JAMA"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"634","DOI":"10.1136\/gutjnl-2017-313952","article-title":"Bacteriophage transfer during faecal microbiota transplantation in Clostridium difficile infection is associated with treatment outcome","volume":"67","author":"Zuo","year":"2018","journal-title":"Gut"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"17562848211017724","DOI":"10.1177\/17562848211017725","article-title":"The contribution of bile acid metabolism to the pathogenesis of Clostridioides difficile infection","volume":"14","author":"Mullish","year":"2021","journal-title":"Therap Adv. Gastroenterol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1556","DOI":"10.1111\/apt.17247","article-title":"Assessing the clinical value of faecal bile acid profiling to predict recurrence in primary Clostridioides difficile infection","volume":"56","author":"Mullish","year":"2022","journal-title":"Aliment. Pharmacol. Ther."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Hughes, J., Aston, C., Kelly, M.L., and Griffin, R. (2022). Towards Development of a Non-Toxigenic Clostridioides difficile Oral Spore Vaccine against Toxigenic C. difficile. Pharmaceutics, 14.","DOI":"10.3390\/pharmaceutics14051086"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Arcay, R., Barcel\u00f3-Nicolau, M., Su\u00e1rez, L., Mart\u00edn, L., Reigada, R., H\u00f6ring, M., Liebisch, G., Garrido, C., Cabot, G., and V\u00edlchez, H. (2024). Gut microbiome and plasma lipidome analysis reveals a specific impact of Clostridioides difficile infection on intestinal bacterial communities and sterol metabolism. mBio, 15.","DOI":"10.1128\/mbio.01347-24"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1902","DOI":"10.1038\/s41591-022-01913-0","article-title":"Drivers and determinants of strain dynamics following fecal microbiota transplantation","volume":"28","author":"Schmidt","year":"2022","journal-title":"Nat. Med."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Deng, Z.L., Pieper, D.H., Stallmach, A., Steube, A., Vital, M., Reck, M., and Wagner-D\u00f6bler, I. (2023). Engraftment of essential functions through multiple fecal microbiota transplants in chronic antibiotic-resistant pouchitis\u2014A case study using metatranscriptomics. Microbiome, 11.","DOI":"10.1186\/s40168-023-01713-9"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1791","DOI":"10.1136\/gutjnl-2018-317842","article-title":"Microbial bile salt hydrolases mediate the efficacy of faecal microbiota transplant in the treatment of recurrent Clostridioides difficile infection","volume":"68","author":"Mullish","year":"2019","journal-title":"Gut"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Lamichhane, S., Sen, P., Alves, M.A., Ribeiro, H.C., Raunioniemi, P., Hy\u00f6tyl\u00e4inen, T., and Ore\u0161i\u010d, M. (2021). Linking Gut Microbiome and Lipid Metabolism: Moving beyond Associations. Metabolites, 11.","DOI":"10.3390\/metabo11010055"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1016\/j.chom.2023.01.009","article-title":"Gut microbiome lipid metabolism and its impact on host physiology","volume":"31","author":"Brown","year":"2023","journal-title":"Cell Host Microbe"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"986","DOI":"10.1038\/s41564-022-01152-6","article-title":"Characterization of inositol lipid metabolism in gut-associated Bacteroidetes","volume":"7","author":"Heaver","year":"2022","journal-title":"Nat. Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Dawkins, J.J., Allegretti, J.R., Gibson, T.E., McClure, E., Delaney, M., Bry, L., and Gerber, G.K. (2022). Gut metabolites predict Clostridioides difficile recurrence. Microbiome, 10.","DOI":"10.1186\/s40168-022-01284-1"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"781","DOI":"10.1038\/s41418-018-0269-2","article-title":"Phosphoinositides: Multipurpose cellular lipids with emerging roles in cell death","volume":"26","author":"Phan","year":"2019","journal-title":"Cell Death Differ."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"12828","DOI":"10.1074\/jbc.M116.773861","article-title":"The natural phosphoinositide derivative glycerophosphoinositol inhibits the lipopolysaccharide-induced inflammatory and thrombotic responses","volume":"292","author":"Vessichelli","year":"2017","journal-title":"J. Biol. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.chom.2017.06.007","article-title":"Indoleacrylic Acid Produced by Commensal Peptostreptococcus Species Suppresses Inflammation","volume":"22","author":"Wlodarska","year":"2017","journal-title":"Cell Host Microbe"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1174","DOI":"10.1136\/gutjnl-2020-323071","article-title":"Gut microbiota-derived metabolites as central regulators in metabolic disorders","volume":"70","author":"Agus","year":"2021","journal-title":"Gut"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e0070624","DOI":"10.1128\/msphere.00706-24","article-title":"Metagenomic, metabolomic, and lipidomic shifts associated with fecal microbiota transplantation for recurrent Clostridioides difficile infection","volume":"9","author":"McMillan","year":"2024","journal-title":"mSphere"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1128\/JB.01492-12","article-title":"Proline-Dependent Regulation of Clostridium difficile Stickland Metabolism","volume":"195","author":"Laurent","year":"2013","journal-title":"J. Bacteriol."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Cersosimo, L.M., Graham, M., Monestier, A., Pavao, A., Worley, J.N., Peltier, J., Dupuy, B., and Bry, L. (2023). Central in vivo mechanisms by which C. difficile\u2019s proline reductase drives efficient metabolism, growth, and toxin production. bioRxiv.","DOI":"10.1101\/2023.05.19.541423"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"770","DOI":"10.1016\/j.chom.2014.11.003","article-title":"Gut Microbiota-Produced Succinate Promotes C. difficile Infection after Antibiotic Treatment or Motility Disturbance","volume":"16","author":"Ferreyra","year":"2014","journal-title":"Cell Host Microbe"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Auria, E., Deschamps, J., Briandet, R., and Dupuy, B. (2023). Extracellular succinate induces spatially organized biofilm formation in Clostridioides difficile. Biofilm, 5.","DOI":"10.1016\/j.bioflm.2023.100125"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Hou, Y., Li, J., and Ying, S. (2023). Tryptophan Metabolism and Gut Microbiota: A Novel Regulatory Axis Integrating the Microbiome, Immunity, and Cancer. Metabolites, 13.","DOI":"10.3390\/metabo13111166"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Neumann-Schaal, M., Jahn, D., and Schmidt-Hohagen, K. (2019). Metabolism the Difficile Way: The Key to the Success of the Pathogen Clostridioides difficile. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.00219"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"716","DOI":"10.1016\/j.chom.2018.05.003","article-title":"Gut Microbiota Regulation of Tryptophan Metabolism in Health and Disease","volume":"23","author":"Agus","year":"2018","journal-title":"Cell Host Microbe"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Antharam, V.C., McEwen, D.C., Garrett, T.J., Dossey, A.T., Li, E.C., Kozlov, A.N., Mesbah, Z., and Wang, G.P. (2016). An Integrated Metabolomic and Microbiome Analysis Identified Specific Gut Microbiota Associated with Fecal Cholesterol and Coprostanol in Clostridium difficile Infection. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0148824"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1038\/s41568-018-0104-6","article-title":"Modelling cancer in microfluidic human organs-on-chips","volume":"19","author":"Hassell","year":"2019","journal-title":"Nat. Rev. Cancer"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e737","DOI":"10.1002\/cpz1.737","article-title":"The Robogut: A Bioreactor Model of the Human Colon for Evaluation of Gut Microbial Community Ecology and Function","volume":"3","author":"Vancuren","year":"2023","journal-title":"Curr. Protoc."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"867","DOI":"10.1016\/j.chom.2021.04.002","article-title":"Organoids and organs-on-chips: Insights into human gut-microbe interactions","volume":"29","author":"Puschhof","year":"2021","journal-title":"Cell Host Microbe"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1038\/s41551-019-0397-0","article-title":"A complex human gut microbiome cultured in an anaerobic intestine-on-a-chip","volume":"3","author":"Gazzaniga","year":"2019","journal-title":"Nat Biomed Eng."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1136\/gutjnl-2023-329515","article-title":"Revisiting the donor screening protocol of faecal microbiota transplantation (FMT): A systematic review","volume":"73","author":"Ng","year":"2024","journal-title":"Gut"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"4703","DOI":"10.1021\/acs.analchem.1c04898","article-title":"Molecular Formula Prediction for Chemical Filtering of 3D OrbiSIMS Datasets","volume":"94","author":"Edney","year":"2022","journal-title":"Anal. Chem."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"van den Berg, R.A., Hoefsloot, H.C.J., Westerhuis, J.A., Smilde, A.K., and van der Werf, M.J. (2006). Centering, scaling, and transformations: Improving the biological information content of metabolomics data. BMC Genom., 7.","DOI":"10.1186\/1471-2164-7-142"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.aca.2013.01.022","article-title":"A consensus orthogonal partial least squares discriminant analysis (OPLS-DA) strategy for multiblock Omics data fusion","volume":"769","author":"Boccard","year":"2013","journal-title":"Anal. Chim. Acta"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Chen, Y., Li, E.-M., and Xu, L.-Y. (2022). Guide to Metabolomics Analysis: A Bioinformatics Workflow. Metabolites, 12.","DOI":"10.3390\/metabo12040357"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Karp, P.D., Paley, S., Caspi, R., Kothari, A., Krummenacker, M., Midford, P.E., Moore, L.R., Subhraveti, P., Gama-Castro, S., and Tierrafria, V.H. (2023). The EcoCyc Database (2023). EcoSal Plus, 11.","DOI":"10.1128\/ecosalplus.esp-0002-2023"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1093\/nar\/27.1.29","article-title":"KEGG: Kyoto Encyclopedia of Genes and Genomes","volume":"27","author":"Ogata","year":"1999","journal-title":"Nucleic Acids Res."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Chong, J., Wishart, D.S., and Xia, J. (2019). Using MetaboAnalyst 4.0 for Comprehensive and Integrative Metabolomics Data Analysis. Curr. Protoc. Bioinform., 68.","DOI":"10.1002\/cpbi.86"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/22\/11221\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,20]],"date-time":"2025-11-20T12:04:17Z","timestamp":1763640257000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/22\/11221"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,20]]},"references-count":51,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["ijms262211221"],"URL":"https:\/\/doi.org\/10.3390\/ijms262211221","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,20]]}}}