{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T19:28:28Z","timestamp":1789759708224,"version":"4.0.1"},"reference-count":61,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2025,4,10]],"date-time":"2025-04-10T00:00:00Z","timestamp":1744243200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Israeli Science Foundation (ISF)","award":["1570\/20"],"award-info":[{"award-number":["1570\/20"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Oral diseases represent a significant global health and economic burden, necessitating the development of effective diagnostic tools. This study investigates the volatile organic compound (VOC) profiles of bacteria associated with dental caries and periodontal disease to explore their potential as diagnostic biomarkers. Four microbial strains\u2014Streptococcus mutans (700610), Streptococcus sanguis (NCO 2863), Porphyromonas gingivalis (ATCC 33277), and Fusobacterium nucleatum (PK1594)\u2014were cultured (N = 24), alongside intraoral samples (N = 60), from individuals with common oral diseases. Headspace VOCs were analyzed using gas chromatography-mass spectrometry (GC-MS), and statistical analyses were conducted by applying non-parametric Wilcoxon and Kruskal\u2013Wallis tests. VOC identification was performed using the NIST14 database. Strain-specific VOC signatures were identified, with P. gingivalis and F. nucleatum exhibiting distinct profiles from each other and from Streptococcus strains. Comparative analysis of disease cohorts revealed statistically significant differences at multiple retention times between caries, gingivitis, and periodontitis. These findings suggest that VOC profiling enables differentiation between bacterial strains and disease phenotypes, supporting their potential application as diagnostic biomarkers for oral diseases. This study establishes a foundational framework for VOC-based diagnostic methodologies in dental pathology.<\/jats:p>","DOI":"10.3390\/ijms26083591","type":"journal-article","created":{"date-parts":[[2025,4,10]],"date-time":"2025-04-10T12:49:01Z","timestamp":1744289341000},"page":"3591","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Bacterial Volatile Organic Compounds as Potential Caries and Periodontitis Disease Biomarkers"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-9970-4222","authenticated-orcid":false,"given":"Maisa","family":"Haiek","sequence":"first","affiliation":[{"name":"Institute of Dental Sciences, Hadassah School of Dental Medicine, The Hebrew University, Jerusalem 9112102, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vladislav","family":"Dvoyris","sequence":"additional","affiliation":[{"name":"Independent Researcher, HaHaroshet St. 12, Or Yehuda 6037580, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0185-2312","authenticated-orcid":false,"given":"Yoav Y.","family":"Broza","sequence":"additional","affiliation":[{"name":"Faculty of Chemical Engineering, Technion and Russell Berrie Nanotechnology Institute\u2014Israel Institute of Technology, Haifa 3200003, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hossam","family":"Haick","sequence":"additional","affiliation":[{"name":"Faculty of Chemical Engineering, Technion and Russell Berrie Nanotechnology Institute\u2014Israel Institute of Technology, Haifa 3200003, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8639-9507","authenticated-orcid":false,"given":"Ervin","family":"Weiss","sequence":"additional","affiliation":[{"name":"The Maurice and Gabriela Goldschleger School of Dental Medicine, Tel-Aviv University, Tel-Aviv 6997801, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yael","family":"Houri-Haddad","sequence":"additional","affiliation":[{"name":"Institute of Dental Sciences, Hadassah School of Dental Medicine, The Hebrew University, Jerusalem 9112102, Israel"},{"name":"Department of Prosthodontics, Hadassah School of Dental Medicine, The Hebrew University, Jerusalem 9112102, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/S0140-6736(19)31146-8","article-title":"Oral diseases: A global public health challenge","volume":"394","author":"Peres","year":"2019","journal-title":"Lancet"},{"key":"ref_2","unstructured":"World Health Organization (2023, May 12). Global Oral Health Status Report: Towards Universal Health Coverage for Oral Health by 2030, Available online: https:\/\/www.who.int\/publications\/i\/item\/9789240061484."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1177\/08959374940080022001","article-title":"Microbial ecology of dental plaque and its significance in health and disease","volume":"8","author":"Marsh","year":"1994","journal-title":"Adv. Dent. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1099\/mic.0.26082-0","article-title":"Are dental diseases examples of ecological catastrophes?","volume":"149","author":"Marsh","year":"2003","journal-title":"Microbiology"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1177\/154411130201300202","article-title":"A mixed-bacteria ecological approach to understanding the role of the oral bacteria in dental caries causation: An alternative to Streptococcus mutans and the specific-plaque hypothesis","volume":"13","author":"Kleinberg","year":"2002","journal-title":"Crit. Rev. Oral Biol. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1159\/000159604","article-title":"Caries ecology revisited: Microbial dynamics and the caries process","volume":"42","author":"Takahashi","year":"2008","journal-title":"Caries Res."},{"key":"ref_7","first-page":"369","article-title":"Microbiology of the early colonization of human enamel and root surfaces in vivo","volume":"95","author":"Nyvad","year":"1987","journal-title":"Scand. J. Dent. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1128\/mr.44.2.331-384.1980","article-title":"Biology, immunology, and cariogenicity of Streptococcus mutans","volume":"44","author":"Hamada","year":"1980","journal-title":"Microbiol. Rev."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1128\/mr.50.4.353-380.1986","article-title":"Role of Streptococcus mutans in human dental decay","volume":"50","author":"Loesche","year":"1986","journal-title":"Microbiol. Rev."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1111\/j.1834-7819.2000.tb00257.x","article-title":"Dental caries is a preventable infectious disease","volume":"45","author":"Balakrishnan","year":"2000","journal-title":"Aust. Dent. J."},{"key":"ref_11","first-page":"5775","article-title":"Relationship between oral microbiota and periodontal disease: A systematic review","volume":"22","author":"Patini","year":"2018","journal-title":"Eur. Rev. Med. Pharmacol. Sci."},{"key":"ref_12","first-page":"7241312","article-title":"Immunological pathways triggered by Porphyromonas gingivalis and Fusobacterium nucleatum: Therapeutic possibilities?","volume":"2019","year":"2019","journal-title":"Mediat. Inflamm."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1111\/j.1600-0757.2005.00113.x","article-title":"Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia: The \u201cred complex\u201d, a prototype polybacterial pathogenic consortium in periodontitis","volume":"38","author":"Holt","year":"2005","journal-title":"Periodontol. 2000"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1046\/j.0906-6713.2002.003421.x","article-title":"Periodontal diagnoses and classification of periodontal diseases","volume":"34","author":"Armitage","year":"2004","journal-title":"Periodontol. 2000"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Baker, J.L., and Edlund, A. (2019). Exploiting the oral microbiome to prevent tooth decay: Has evolution already provided the best tools?. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.03323"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Sedghi, L.M., Bacino, M., and Kapila, Y.L. (2021). Periodontal disease: The good, the bad, and the unknown. Front. Cell. Infect. Microbiol., 11.","DOI":"10.3389\/fcimb.2021.766944"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"024001","DOI":"10.1088\/1752-7155\/6\/2\/024001","article-title":"Microbial volatile compounds in health and disease conditions","volume":"6","author":"Thorn","year":"2012","journal-title":"J. Breath Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"11761","DOI":"10.1021\/acs.chemrev.9b00437","article-title":"Disease detection with molecular biomarkers: From chemistry of body fluids to nature-inspired chemical sensors","volume":"119","author":"Broza","year":"2019","journal-title":"Chem. Rev."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1021\/acsnano.6b04930","article-title":"Diagnosis and classification of 17 diseases from 1404 subjects via pattern analysis of exhaled molecules","volume":"11","author":"Nakhleh","year":"2017","journal-title":"ACS Nano"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"814","DOI":"10.1039\/b507392h","article-title":"Bacterial volatiles: The smell of small organisms","volume":"24","author":"Schulz","year":"2007","journal-title":"Nat. Prod. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"785","DOI":"10.2217\/nnm.13.64","article-title":"Nanomaterial-based sensors for detection of disease by volatile organic compounds","volume":"8","author":"Broza","year":"2013","journal-title":"Nanomedicine"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1146\/annurev-micro-011320-015542","article-title":"Bacterial Volatile Compounds: Functions in Communication, Cooperation, and Competition","volume":"74","author":"Netzker","year":"2020","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"8185","DOI":"10.1007\/s12274-022-4459-3","article-title":"Volatolomics in healthcare and its advanced detection technology","volume":"15","author":"Hu","year":"2022","journal-title":"Nano Res."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Serasanambati, M., Broza, Y.Y., and Haick, H. (2019). Volatile Compounds Are Involved in Cellular Crosstalk and Upregulation. Adv. Biosyst., 3.","DOI":"10.1002\/adbi.201900131"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1111\/j.1600-0765.1992.tb01673.x","article-title":"Volatile sulfur compounds in mouth air from clinically healthy subjects and patients with periodontal disease","volume":"27","author":"Yaegaki","year":"1992","journal-title":"J. Periodontal Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/S0378-4347(99)00127-9","article-title":"Variation in volatile organic compounds in the breath of normal humans","volume":"729","author":"Phillips","year":"1999","journal-title":"J. Chromatogr. B Biomed. Sci. Appl."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1111\/j.1601-0825.2005.01085.x","article-title":"Pilot study of a breath test for volatile organic compounds associated with oral malodor: Evidence for the role of oxidative stress","volume":"11","author":"Phillips","year":"2005","journal-title":"Oral Diseases"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.jchromb.2007.02.048","article-title":"Halitosis-associated volatiles in breath of healthy subjects","volume":"853","author":"Quirynen","year":"2007","journal-title":"J. Chromatogr. B Anal. Technol. Biomed. Life Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.14219\/jada.archive.2007.0325","article-title":"Identification of oral bacterial species associated with halitosis","volume":"138","author":"Haraszthy","year":"2007","journal-title":"J. Am. Dent. Assoc."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1177\/0022034508329741","article-title":"Detection of odorous compounds in breath","volume":"88","author":"Quirynen","year":"2009","journal-title":"J. Dent. Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1080\/10426507.2012.736894","article-title":"Volatile Sulfur Compounds as the Cause of Bad Breath: A Review","volume":"188","author":"Tangerman","year":"2013","journal-title":"Phosphorus Sulfur Silicon Relat. Elem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"017114","DOI":"10.1088\/1752-7155\/7\/1\/017114","article-title":"Volatiles from oral anaerobes confounding breath biomarker discovery","volume":"7","author":"Khalid","year":"2013","journal-title":"J. Breath Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1111\/odi.12699","article-title":"Detection of halitosis in breath: Between the past, present, and future","volume":"24","author":"Nakhleh","year":"2018","journal-title":"Oral Dis."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Roslund, K., Uosukainen, M., J\u00e4rvik, K., Hartonen, K., Lehto, M., Pussinen, P., Groop, P.H., and Mets\u00e4l\u00e4, M. (2022). Antibiotic treatment and supplemental hemin availability affect the volatile organic compounds produced by P. gingivalis in vitro. Sci. Rep., 12.","DOI":"10.1038\/s41598-022-26497-0"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1007\/s10096-015-2536-1","article-title":"Detection of signature volatiles for cariogenic microorganisms","volume":"35","author":"Hertel","year":"2016","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1007\/s00784-017-2147-6","article-title":"Volatile organic compounds in the breath of oral candidiasis patients: A pilot study","volume":"22","author":"Hertel","year":"2018","journal-title":"Clin. Oral Investig."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"016010","DOI":"10.1088\/1752-7163\/ab5559","article-title":"On-line profiling of volatile compounds produced in vitro by pathogenic oral bacteria","volume":"14","author":"Roslund","year":"2019","journal-title":"J. Breath Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1002\/hed.26043","article-title":"Exhaled breath analysis in the diagnosis of head and neck cancer","volume":"42","author":"Almangush","year":"2020","journal-title":"Head Neck"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Mentel, S., Gallo, K., Wagendorf, O., Preissner, R., Nahles, S., Heiland, M., and Preissner, S. (2021). Prediction of oral squamous cell carcinoma based on machine learning of breath samples: A prospective controlled study. BMC Oral Health, 21.","DOI":"10.1186\/s12903-021-01862-z"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Roslund, K., Lehto, M., Pussinen, P., Hartonen, K., Groop, P.H., Halonen, L., and Mets\u00e4l\u00e4, M. (2021). Identifying volatile in vitro biomarkers for oral bacteria with proton-transfer-reaction mass spectrometry and gas chromatography-mass spectrometry. Sci. Rep., 11.","DOI":"10.1038\/s41598-021-96287-7"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1111\/j.1600-0757.2011.00436.x","article-title":"Examiner alignment and assessment in clinical periodontal research","volume":"59","author":"Hefti","year":"2012","journal-title":"Periodontol. 2000"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1111\/jcpe.13104","article-title":"Implementation of the new classification of periodontal diseases: Decision-making algorithms for clinical practice and education","volume":"46","author":"Tonetti","year":"2019","journal-title":"J. Clin. Periodontol."},{"key":"ref_43","unstructured":"National Center for Biotechnology Information (2023, December 19). PubChem Compound Summary for CID 6433417, 2-Methyl-2-buten-1-OL, Available online: https:\/\/pubchem.ncbi.nlm.nih.gov\/compound\/2-Methyl-2-buten-1-OL."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Magrin, G.L., Strauss, F.J., Benfatti, C.A.M., Maia, L.C., and Gruber, R. (2020). Effects of short-chain fatty acids on human oral epithelial cells and the potential impact on periodontal disease: A systematic review of in vitro studies. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21144895"},{"key":"ref_45","unstructured":"National Center for Biotechnology Information (2023, December 19). PubChem Compound Summary for CID 141153, 2,4-Dimethyl-2,4-pentanediol, Available online: https:\/\/pubchem.ncbi.nlm.nih.gov\/compound\/141153."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1775","DOI":"10.1038\/s41416-020-01051-9","article-title":"Development of a non-invasive exhaled breath test for the diagnosis of head and neck cancer","volume":"123","author":"Dharmawardana","year":"2020","journal-title":"Br. J. Cancer"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.ijms.2007.05.012","article-title":"Lung cancer detection by proton transfer reaction mass-spectrometric analysis of human breath gas","volume":"265","author":"Wehinger","year":"2007","journal-title":"Int. J. Mass Spectrom."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1227","DOI":"10.1111\/j.1462-2920.2005.00805.x","article-title":"Isolation and molecular detection of methylotrophic bacteria occurring in the human mouth","volume":"7","author":"Anesti","year":"2005","journal-title":"Environ. Microbiol."},{"key":"ref_49","first-page":"193","article-title":"\u201cICDAS\u201d\u2014An international system for caries detection and assessment being developed to facilitate caries epidemiology, research, and appropriate clinical management","volume":"21","author":"Pitts","year":"2004","journal-title":"Community Dent. Health"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1038\/nature23481","article-title":"Mammals divert endogenous genotoxic formaldehyde into one-carbon metabolism","volume":"548","author":"Wit","year":"2017","journal-title":"Nature"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.molmet.2019.05.012","article-title":"Formate metabolism in health and disease","volume":"33","author":"Pietzke","year":"2019","journal-title":"Mol. Metab."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1902\/jop.1999.70.5.485","article-title":"The relationship between oral malodor, gingivitis, and periodontitis: A review","volume":"70","author":"Ratcliff","year":"1999","journal-title":"J. Periodontol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1111\/j.1399-302X.1997.tb00733.x","article-title":"Acid tolerance and acid-neutralizing activity of Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum","volume":"12","author":"Takahashi","year":"1997","journal-title":"Oral Microbiol. Immunol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1158\/1541-7786.MCR-05-0126","article-title":"Evidence that hydrogen sulfide is a genotoxic agent","volume":"4","author":"Wagner","year":"2006","journal-title":"Mol. Cancer Res."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Karpi\u0144ski, T.M. (2019). Role of Oral Microbiota in Cancer Development. Microorganisms, 7.","DOI":"10.3390\/microorganisms7010020"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"4466","DOI":"10.1128\/JVI.03326-13","article-title":"Short-chain fatty acids from periodontal pathogens suppress histone deacetylases, EZH2, and SUV39H1 to promote Kaposi\u2019s sarcoma-associated herpesvirus replication","volume":"88","author":"Yu","year":"2014","journal-title":"J. Virol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.tifs.2020.02.026","article-title":"Dynamic balancing of intestinal short-chain fatty acids: The crucial role of bacterial metabolism","volume":"100","author":"Xu","year":"2020","journal-title":"Trends Food Sci. Technol."},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Leonov, G.E., Varaeva, Y.R., Livantsova, E.N., and Starodubova, A.V. (2023). The complicated relationship of short-chain fatty acids and oral microbiome: A narrative review. Biomedicines, 11.","DOI":"10.3390\/biomedicines11102749"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1499","DOI":"10.1021\/ed078p1499","article-title":"The Henderson-Hasselbalch equation: Its history and limitations","volume":"78","author":"Po","year":"2001","journal-title":"J. Chem. Educ."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1080\/10473289.1997.10464411","article-title":"Monitoring VOCs in Air Using Sorbent Tubes Followed by Thermal Desorption-Capillary GC Analysis: Summary of Data and Practical Guidelines","volume":"47","author":"Woolfenden","year":"1997","journal-title":"J. Air Waste Manag. Assoc."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"3047","DOI":"10.1111\/j.1462-2920.2011.02582.x","article-title":"Production of plant growth modulating volatiles is widespread among rhizosphere bacteria and strongly depends on culture conditions","volume":"13","author":"Blom","year":"2011","journal-title":"Environ. Microbiol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/8\/3591\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:12:31Z","timestamp":1760029951000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/8\/3591"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,10]]},"references-count":61,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2025,4]]}},"alternative-id":["ijms26083591"],"URL":"https:\/\/doi.org\/10.3390\/ijms26083591","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,10]]}}}