{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:45:56Z","timestamp":1760240756597,"version":"build-2065373602"},"reference-count":38,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2019,8,29]],"date-time":"2019-08-29T00:00:00Z","timestamp":1567036800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100013704","name":"CONACYT","doi-asserted-by":"publisher","award":["FOSISS-261569"],"award-info":[{"award-number":["FOSISS-261569"]}],"id":[{"id":"10.13039\/501100013704","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100013704","name":"CONACyT","doi-asserted-by":"publisher","award":["CB-221137"],"award-info":[{"award-number":["CB-221137"]}],"id":[{"id":"10.13039\/501100013704","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Non-alcoholic fatty liver disease is a highly prevalent condition worldwide that increases the risk to develop liver fibrosis, cirrhosis, and hepatocellular carcinoma. Thus, it is imperative to develop novel diagnostic tools that together with liver biopsy help to differentiate mild and advanced degrees of steatosis. Ex-vivo liver samples were collected from mice fed a methionine-choline deficient diet for two or eight weeks, and from a control group. The degree of hepatic steatosis was histologically evaluated, and fat content was assessed by Oil-Red O staining. On the other hand, fluorescence spectroscopy was used for the assessment of the steatosis progression. Fluorescence spectra were recorded at excitation wavelengths of 330, 365, 385, 405, and 415 nm by establishing surface contact of the fiber optic probe with the liver specimens. A multi-variate statistical approach based on principal component analysis followed by quadratic discriminant analysis was applied to spectral data to obtain classifiers able to distinguish mild and moderate stages of steatosis at the different excitation wavelengths. Receiver Operating Characteristic (ROC) curves were computed to compare classifier\u2019s performances for each one of the five excitation wavelengths and steatosis stages. Optimal sensitivity and specificity were calculated from the corresponding ROC curves using the Youden index. Intensity in the endogenous fluorescence spectra at the given wavelengths progressively increased according to the time of exposure to diet. The area under the curve of the spectra was able to discriminate control liver samples from those with steatosis and differentiate among the time of exposure to the diet for most of the used excitation wavelengths. High specificities and sensitivities were obtained for every case; however, fluorescence spectra obtained by exciting with 405 nm yielded the best results distinguishing between the mentioned classes with a total classification error of 1.5% and optimal sensitivities and specificities better than 98.6% and 99.3%, respectively.<\/jats:p>","DOI":"10.3390\/molecules24173150","type":"journal-article","created":{"date-parts":[[2019,8,29]],"date-time":"2019-08-29T11:26:22Z","timestamp":1567077982000},"page":"3150","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Study of Methionine Choline Deficient Diet-Induced Steatosis in Mice Using Endogenous Fluorescence Spectroscopy"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6648-7905","authenticated-orcid":false,"given":"Alma","family":"Valor","sequence":"first","affiliation":[{"name":"Laboratorio de Biofot\u00f3nica, ESIME Zac, Instituto Polit\u00e9cnico Nacional, Ciudad de Mexico 07738, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2261-771X","authenticated-orcid":false,"given":"Eduardo J.","family":"Arista Romeu","sequence":"additional","affiliation":[{"name":"Laboratorio de Biofot\u00f3nica, ESIME Zac, Instituto Polit\u00e9cnico Nacional, Ciudad de Mexico 07738, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Galileo","family":"Escobedo","sequence":"additional","affiliation":[{"name":"Laboratorio de Prote\u00f3mica, Direcci\u00f3n de Investigaci\u00f3n, Hospital General de Mexico \u201cDr. Eduardo Liceaga\u201d, Ciudad de Mexico 06720, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adriana","family":"Campos-Espinosa","sequence":"additional","affiliation":[{"name":"Laboratorio de H\u00edgado, P\u00e1ncreas y Motilidad, Unidad de Medicina Experimental, Facultad de Medicina, Universidad Nacional Aut\u00f3noma de Mexico\/Hospital General de Mexico \u201cDr. Eduardo Liceaga\u201d, Ciudad de Mexico 06720, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ivette Irais","family":"Romero-Bello","sequence":"additional","affiliation":[{"name":"Laboratorio de H\u00edgado, P\u00e1ncreas y Motilidad, Unidad de Medicina Experimental, Facultad de Medicina, Universidad Nacional Aut\u00f3noma de Mexico\/Hospital General de Mexico \u201cDr. Eduardo Liceaga\u201d, Ciudad de Mexico 06720, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Javier","family":"Moreno-Gonz\u00e1lez","sequence":"additional","affiliation":[{"name":"Laboratorio de H\u00edgado, P\u00e1ncreas y Motilidad, Unidad de Medicina Experimental, Facultad de Medicina, Universidad Nacional Aut\u00f3noma de Mexico\/Hospital General de Mexico \u201cDr. Eduardo Liceaga\u201d, Ciudad de Mexico 06720, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6383-1675","authenticated-orcid":false,"given":"Diego A.","family":"Fabila Bustos","sequence":"additional","affiliation":[{"name":"Laboratorio de Biofot\u00f3nica, ESIME Zac, Instituto Polit\u00e9cnico Nacional, Ciudad de Mexico 07738, Mexico"},{"name":"Laboratorio de Espectroscopia, UPIIH, Instituto Polit\u00e9cnico Nacional, Ciudad del Conocimiento y la Cultura, San Agust\u00edn Tlaxiaca 42162, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Suren","family":"Stolik","sequence":"additional","affiliation":[{"name":"Laboratorio de Biofot\u00f3nica, ESIME Zac, Instituto Polit\u00e9cnico Nacional, Ciudad de Mexico 07738, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jose Manuel","family":"de la Rosa V\u00e1zquez","sequence":"additional","affiliation":[{"name":"Laboratorio de Biofot\u00f3nica, ESIME Zac, Instituto Polit\u00e9cnico Nacional, Ciudad de Mexico 07738, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carolina","family":"Guzm\u00e1n","sequence":"additional","affiliation":[{"name":"Laboratorio de H\u00edgado, P\u00e1ncreas y Motilidad, Unidad de Medicina Experimental, Facultad de Medicina, Universidad Nacional Aut\u00f3noma de Mexico\/Hospital General de Mexico \u201cDr. Eduardo Liceaga\u201d, Ciudad de Mexico 06720, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,8,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1016\/j.dld.2014.09.020","article-title":"Nonalcoholic fatty liver disease: A precursor of the metabolic syndrome","volume":"47","author":"Lonardo","year":"2015","journal-title":"Dig. Liver Dis."},{"doi-asserted-by":"crossref","unstructured":"Calzadilla Bertot, L., and Adams, L.A. (2016). The natural course of non-alcoholic fatty liver disease. Int. J. Mol. Sci., 17.","key":"ref_2","DOI":"10.3390\/ijms17050774"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1007\/s12072-013-9446-z","article-title":"Current histological classification of NAFLD: Strength and limitations","volume":"7","author":"Bedossa","year":"2013","journal-title":"Hepatol. Int."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"526979","DOI":"10.1155\/2014\/526979","article-title":"Nonalcoholic fatty liver disease: Different classifications concordance and relationship between degrees of morphological features and spectrum of the disease","volume":"2014","author":"Monteiro","year":"2014","journal-title":"Anal. Cell. Pathol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e15167","DOI":"10.5812\/hepatmon.15167","article-title":"Inter-observer and intra-observer agreement in pathological evaluation of non-alcoholic fatty liver disease suspected liver biopsies","volume":"14","author":"Pournik","year":"2014","journal-title":"Hepat. Mon."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"877","DOI":"10.1111\/apt.13770","article-title":"Diagnostic performance of FibroTest, SteatoTest and ActiTest in patients with NAFLD using the SAF score as histological reference","volume":"44","author":"Munteanu","year":"2016","journal-title":"Aliment. Pharmacol. Ther."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1357","DOI":"10.1366\/14-07462","article-title":"Diffuse reflectance spectroscopy as a possible tool to complement liver biopsy for grading hepatic fibrosis in paraffin-preserved human liver specimens","volume":"68","author":"Brown","year":"2014","journal-title":"Appl. Spectrosc."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1366\/10-06052","article-title":"Portable, fiber-based, diffuse reflection spectroscopy (DRS) systems for estimating tissue optical properties","volume":"65","author":"Vishwanath","year":"2011","journal-title":"Appl. Spectrosc."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1562\/0031-8655(2003)078<0384:MFEMRT>2.0.CO;2","article-title":"Molecular fluorescence excitation\u2013emission matrices relevant to tissue spectroscopy","volume":"78","author":"Andersson","year":"2003","journal-title":"Photochem. Photobiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1016\/j.pdpdt.2012.05.002","article-title":"In vivo assessment of liver fibrosis using diffuse reflectance and fluorescence spectroscopy: a proof of concept","volume":"9","author":"Fabila","year":"2012","journal-title":"Photodiagn. Photodyn. Ther."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1898","DOI":"10.1053\/j.gastro.2005.03.084","article-title":"Sampling variability of liver biopsy in nonalcoholic fatty liver disease","volume":"128","author":"Ratziu","year":"2005","journal-title":"Gastroenterology"},{"key":"ref_12","first-page":"106854P","article-title":"Application of the fluorescence spectroscopy for the analysis of the state of abdominal cavity organs tissues in mini-invasive surgery","volume":"Volume 10685","author":"Kandurova","year":"2018","journal-title":"Biophotonics: Photonic Solutions for Better Health Care VI, Proceedings of the SPIE 10685 Photonics Europe International Society for Optics and Photonics, Strasbourg, France, 17 May 2018"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/S1387-2656(05)11007-2","article-title":"Cell and tissue autofluorescence research and diagnostic applications","volume":"11","author":"Monici","year":"2005","journal-title":"Biotechnol. Annu. Rev."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1160","DOI":"10.1111\/liv.13753","article-title":"Autofluorescence-based optical biopsy: An effective diagnostic tool in hepatology","volume":"38","author":"Croce","year":"2018","journal-title":"Liver Int."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"602","DOI":"10.1097\/SLA.0000000000001452","article-title":"The optical biopsy: A novel technique for rapid intraoperative diagnosis of primary pulmonary adenocarcinomas","volume":"262","author":"Kennedy","year":"2015","journal-title":"Ann. Surg."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1002\/lsm.20923","article-title":"Human liver autofluorescence: An intrinsic tissue parameter discriminating normal and diseased conditions","volume":"42","author":"Croce","year":"2010","journal-title":"Lasers Surg. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"803491","DOI":"10.1155\/2014\/803491","article-title":"Integrated autofluorescence characterization of a modified-diet liver model with accumulation of lipids and oxidative stress","volume":"2014","author":"Croce","year":"2014","journal-title":"Biomed. Res. Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1111\/j.1478-3231.2008.01878.x","article-title":"Fluorescence spectroscopy to diagnose hepatic steatosis in a rat model of fatty liver","volume":"29","author":"Sankarankutty","year":"2009","journal-title":"Liver Int."},{"key":"ref_19","first-page":"1","article-title":"Diffuse reflectance spectroscopy accurately discriminates early and advanced grades of fatty liver in mice","volume":"23","author":"Escobedo","year":"2018","journal-title":"J. Biomed. Opt."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1590\/S0004-28032011000100015","article-title":"Methionine- and choline-deficient diet induces hepatic changes characteristic of non-alcoholic steatohepatitis","volume":"48","author":"Marcolin","year":"2011","journal-title":"Arq. Gastroenterol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1002\/lsm.21064","article-title":"Multi-excitation fluorescence spectroscopy for analysis of non-alcoholic fatty liver disease","volume":"43","author":"Sauvage","year":"2011","journal-title":"Lasers Surg. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2325","DOI":"10.1364\/AO.28.002325","article-title":"Optical properties of rat liver between 350 and 2200 nm","volume":"28","author":"Parsa","year":"1989","journal-title":"Appl. Opt."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1070\/QEL16903","article-title":"Measuring optical properties of human liver between 400 and 1000 nm","volume":"49","author":"Carneiro","year":"2019","journal-title":"Quantum Electron."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1042\/bj0220987","article-title":"The absorption spectrum of vitamin A","volume":"22","author":"Morton","year":"1928","journal-title":"Biochem. J."},{"doi-asserted-by":"crossref","unstructured":"Saeed, A., Dullaart, R.P.F., Schreuder, T.C.M.A., Blokzijl, H., and Faber, K.N. (2018). Disturbed vitamin A metabolism in Non-Alcoholic Fatty Liver Disease (NAFLD). Nutrients, 10.","key":"ref_25","DOI":"10.3390\/nu10010029"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"920","DOI":"10.1039\/b407358d","article-title":"Autofluorescence properties of isolated rat hepatocytes under different metabolic conditions","volume":"3","author":"Croce","year":"2004","journal-title":"Photochem. Photobiol. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.jphotobiol.2016.09.015","article-title":"Autofluorescence discrimination of metabolic fingerprint in nutritional and genetic fatty liver models","volume":"164","author":"Croce","year":"2016","journal-title":"J. Photochem. Photobiol. B"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"S819","DOI":"10.1016\/S0168-8278(18)31911-1","article-title":"Lipofuscin is detected in early stages of the disease in liver biopsies of patients with Non-Alcoholic Fatty Liver Disease","volume":"68","author":"Morrison","year":"2018","journal-title":"J. Hepatol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2576","DOI":"10.4081\/ejh.2015.2576","article-title":"New light in flavin autofluorescence","volume":"59","author":"Croce","year":"2015","journal-title":"Eur. J. Histochem."},{"key":"ref_30","first-page":"2485","article-title":"Lipofuscin, lipofuscin-like pigments and autofluorescence","volume":"59","year":"2015","journal-title":"Eur. J. Histochem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"10967","DOI":"10.1038\/s41598-018-29370-1","article-title":"Fluorescence spectroscopy as an efficient tool for staging the degree of liver fibrosis: an in vivo comparison with MRI","volume":"8","author":"Nazeer","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1313","DOI":"10.1002\/hep.20701","article-title":"Design and validation of a histological scoring system for nonalcoholic fatty liver disease","volume":"41","author":"Kleiner","year":"2005","journal-title":"Hepatology"},{"unstructured":"Fabila, D.A. (2010). Desarrollo de un espectrofluor\u00f3metro port\u00e1til para mediciones in situ de tejido biol\u00f3gico. [Master\u2019s Thesis, Instituto Politecnico Nacional]. Available online: https:\/\/www.sepi.esimez.ipn.mx\/electronica\/archivos\/742.pdf.","key":"ref_33"},{"unstructured":"Miller, J.N., and Miller, J.C. (2010). Statistics and chemometrics for analytical chemistry, Pearson Education. [6th ed.].","key":"ref_34"},{"doi-asserted-by":"crossref","unstructured":"Hastie, T., Tibshirani, R., and Friedman, J. (2009). The Elements of Statistical Learning: Data Mining; Inference; and Prediction, Springer. [2nd ed.].","key":"ref_35","DOI":"10.1007\/978-0-387-84858-7"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.chemolab.2008.07.010","article-title":"Comparison of performance of five common classifiers represented as boundary methods: Euclidean distance to centroids, linear discriminant analysis, quadratic discriminant analysis, learning vector quantization and support vector machines, as dependent on data structure","volume":"95","author":"Dixon","year":"2009","journal-title":"Chemom. Intell. Lab. Syst."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1007\/s13312-011-0055-4","article-title":"Receiver operating characteristic (ROC) curve for medical researchers","volume":"48","author":"Kumar","year":"2011","journal-title":"Indian pediatr."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1002\/bimj.200710415","article-title":"Youden index and optimal cut-point estimated from observations affected by a lower limit of detection","volume":"50","author":"Ruopp","year":"2008","journal-title":"Biom. J."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/24\/17\/3150\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:15:02Z","timestamp":1760188502000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/24\/17\/3150"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,8,29]]},"references-count":38,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2019,9]]}},"alternative-id":["molecules24173150"],"URL":"https:\/\/doi.org\/10.3390\/molecules24173150","relation":{},"ISSN":["1420-3049"],"issn-type":[{"type":"electronic","value":"1420-3049"}],"subject":[],"published":{"date-parts":[[2019,8,29]]}}}