{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T18:08:18Z","timestamp":1784225298883,"version":"3.55.0"},"reference-count":40,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T00:00:00Z","timestamp":1784160000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003627","name":"Rural Development Administration","doi-asserted-by":"publisher","award":["PJ01398501"],"award-info":[{"award-number":["PJ01398501"]}],"id":[{"id":"10.13039\/501100003627","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    Canine brucellosis caused by\n                    <jats:italic>Brucella canis<\/jats:italic>\n                    is gaining public-health relevance as companion-dog ownership and close human\u2013animal contact increase. Yet mechanistic insight into\n                    <jats:italic>B. canis<\/jats:italic>\n                    host responses remains limited at the pathway level. To address this gap, we profiled infection-associated metabolic responses in RAW 264.7 cells across multiple post-infection time points using targeted metabolomics (CE\u2013MS). We observed a coherent, time-ordered pattern with the largest shifts at 24 h: arginine, hydroxyproline, choline, cysteine, \u03b2-alanine, and glycerol-3-phosphate showed pronounced decreases, and tyrosine decreased modestly; \u03b1-ketoglutarate displayed a lower mean at 12 h. Unsupervised analyses separated infected and control groups over time (PCA PC1\u2009=\u200959.4%, PC2\u2009=\u200916.7%; 76.1% cumulative), providing an orthogonal summary of the temporal divergence. A supplementary sensitivity analysis (Minimum Cell Loss) was included as exploratory contextual information regarding potential cell-number effects, although direct measurements of infection efficiency and macrophage viability were not performed. An integrated pathway schematic summarizes metabolite associations related to nitric-oxide\u2013related, mitochondrial\/energy, redox\/glutathione, extracellular-matrix, membrane-lipid, and neuroimmune-related pathways. Together, these results deliver a time-resolved view of\n                    <jats:italic>B. canis<\/jats:italic>\n                    \u2013macrophage metabolic responses and delineate prioritized avenues for targeted mechanistic validation.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0353470","type":"journal-article","created":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T17:48:41Z","timestamp":1784224121000},"page":"e0353470","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Targeted metabolomic profiling of RAW 264.7 cells infected with Brucella canis identifies time-dependent metabolic changes"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6893-0499","authenticated-orcid":true,"given":"Woo Bin","family":"Park","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Su Min","family":"Kyung","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8901-0085","authenticated-orcid":true,"given":"Suji","family":"Kim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Young Ju","family":"Lee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0548-0835","authenticated-orcid":true,"given":"Han Sang","family":"Yoo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,7,16]]},"reference":[{"key":"pone.0353470.ref001","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1146\/annurev-micro-090110-102905","article-title":"Interactions of the human pathogenic Brucella species with their hosts","volume":"65","author":"VL Atluri","year":"2011","journal-title":"Annu Rev Microbiol"},{"issue":"3","key":"pone.0353470.ref002","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1111\/tbed.12632","article-title":"Brucella canis infection in dogs from commercial breeding kennels in Brazil","volume":"64","author":"LB Keid","year":"2017","journal-title":"Transbound Emerg Dis"},{"issue":"2","key":"pone.0353470.ref003","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1017\/S0950268809990525","article-title":"Human Brucella canis outbreak linked to infection in dogs","volume":"138","author":"NE Lucero","year":"2010","journal-title":"Epidemiol Infect"},{"key":"pone.0353470.ref004","volume-title":"Brucellosis in humans and animals","author":"MJ Corbel","year":"2006"},{"key":"pone.0353470.ref005","article-title":"Global burden of human brucellosis: A systematic review","volume":"6","author":"AS Dean","year":"2012","journal-title":"PLoS Negl Trop Dis"},{"issue":"4","key":"pone.0353470.ref006","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1007\/s00430-009-0123-8","article-title":"Survival of the fittest: how Brucella strains adapt to their intracellular niche in the host","volume":"198","author":"2nd Roop RM","year":"2009","journal-title":"Med Microbiol Immunol"},{"issue":"3","key":"pone.0353470.ref007","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1111\/j.1574-6976.2012.00334.x","article-title":"Internal affairs: investigating the Brucella intracellular lifestyle","volume":"36","author":"K von Bargen","year":"2012","journal-title":"FEMS Microbiol Rev"},{"issue":"8","key":"pone.0353470.ref008","doi-asserted-by":"crossref","first-page":"1401","DOI":"10.3201\/eid2408.171171","article-title":"Brucellosis in Dogs and Public Health Risk","volume":"24","author":"ME Hensel","year":"2018","journal-title":"Emerg Infect Dis"},{"key":"pone.0353470.ref009","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1016\/j.vetmic.2009.06.021","article-title":"Brucellosis: A re-emerging zoonosis","volume":"140","author":"MN Seleem","year":"2010","journal-title":"Vet Microbiol"},{"issue":"1","key":"pone.0353470.ref010","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1111\/imr.12266","article-title":"Macrophage defense mechanisms against intracellular bacteria","volume":"264","author":"G Weiss","year":"2015","journal-title":"Immunol Rev"},{"key":"pone.0353470.ref011","doi-asserted-by":"crossref","first-page":"24","DOI":"10.3389\/fcimb.2013.00024","article-title":"Metabolic host responses to infection by intracellular bacterial pathogens","volume":"3","author":"W Eisenreich","year":"2013","journal-title":"Front Cell Infect Microbiol"},{"issue":"3","key":"pone.0353470.ref012","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0194126","article-title":"Metabolomic profiling of macrophages determines the discrete metabolomic signature and metabolomic interactome triggered by polarising immune stimuli","volume":"13","author":"KM Rattigan","year":"2018","journal-title":"PLoS One"},{"issue":"4","key":"pone.0353470.ref013","doi-asserted-by":"crossref","first-page":"1819","DOI":"10.1152\/physrev.00035.2018","article-title":"Metabolomics for Investigating Physiological and Pathophysiological Processes","volume":"99","author":"DS Wishart","year":"2019","journal-title":"Physiol Rev"},{"issue":"6","key":"pone.0353470.ref014","doi-asserted-by":"crossref","first-page":"108","DOI":"10.3390\/metabo9060108","article-title":"Translational Metabolomics: Current Challenges and Future Opportunities","volume":"9","author":"FR Pinu","year":"2019","journal-title":"Metabolites"},{"key":"pone.0353470.ref015","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1258\/000456303321016132","article-title":"Proton nuclear magnetic resonance spectroscopy of body fluids in the field of inborn errors of metabolism","volume":"40","author":"SH Moolenaar","year":"2003","journal-title":"Ann Clin Biochem"},{"issue":"17","key":"pone.0353470.ref016","doi-asserted-by":"crossref","DOI":"10.1128\/JB.00128-18","article-title":"Reduce, Induce, Thrive: Bacterial Redox Sensing during Pathogenesis","volume":"200","author":"ML Reniere","year":"2018","journal-title":"J Bacteriol"},{"key":"pone.0353470.ref017","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.coi.2019.05.009","article-title":"Metabolic reprogramming: an innate cellular defence mechanism against intracellular bacteria?","volume":"60","author":"P Escoll","year":"2019","journal-title":"Curr Opin Immunol"},{"issue":"5","key":"pone.0353470.ref018","doi-asserted-by":"crossref","first-page":"2238","DOI":"10.1021\/pr101054m","article-title":"(1)H NMR-based metabolomic profiling in mice infected with Mycobacterium tuberculosis","volume":"10","author":"J-H Shin","year":"2011","journal-title":"J Proteome Res"},{"key":"pone.0353470.ref019","doi-asserted-by":"crossref","first-page":"619759","DOI":"10.3389\/fvets.2021.619759","article-title":"Identification of Dendritic Cell Maturation, TLR, and TREM1 Signaling Pathways in the Brucella canis Infected Canine Macrophage Cells, DH82, Through Transcriptomic Analysis","volume":"8","author":"WB Park","year":"2021","journal-title":"Front Vet Sci"},{"issue":"5","key":"pone.0353470.ref020","doi-asserted-by":"crossref","first-page":"577","DOI":"10.3390\/antiox13050577","article-title":"Brucella Manipulates Host Cell Ferroptosis to Facilitate Its Intracellular Replication and Egress in RAW264.7 Macrophages","volume":"13","author":"G Zhang","year":"2024","journal-title":"Antioxidants (Basel)"},{"key":"pone.0353470.ref021","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.mam.2008.08.006","article-title":"Glutathione: overview of its protective roles, measurement, and biosynthesis","volume":"30","author":"HJ Forman","year":"2009","journal-title":"Mol Aspects Med"},{"issue":"1","key":"pone.0353470.ref022","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1186\/s41232-024-00352-3","article-title":"Peripheral and central regulation of neuro-immune crosstalk","volume":"44","author":"M Izumi","year":"2024","journal-title":"Inflamm Regen"},{"issue":"6","key":"pone.0353470.ref023","doi-asserted-by":"crossref","first-page":"1236","DOI":"10.1021\/ac990976y","article-title":"Amino acid analysis by capillary electrophoresis electrospray ionization mass spectrometry","volume":"72","author":"T Soga","year":"2000","journal-title":"Anal Chem"},{"issue":"10","key":"pone.0353470.ref024","doi-asserted-by":"crossref","first-page":"2233","DOI":"10.1021\/ac020064n","article-title":"Simultaneous determination of anionic intermediates for Bacillus subtilis metabolic pathways by capillary electrophoresis electrospray ionization mass spectrometry","volume":"74","author":"T Soga","year":"2002","journal-title":"Anal Chem"},{"issue":"5","key":"pone.0353470.ref025","doi-asserted-by":"crossref","first-page":"488","DOI":"10.1021\/pr034020m","article-title":"Quantitative metabolome analysis using capillary electrophoresis mass spectrometry","volume":"2","author":"T Soga","year":"2003","journal-title":"J Proteome Res"},{"issue":"24","key":"pone.0353470.ref026","doi-asserted-by":"crossref","first-page":"21123","DOI":"10.1074\/jbc.M110675200","article-title":"Regulation of T cell receptor CD3zeta chain expression by L-arginine","volume":"277","author":"PC Rodriguez","year":"2002","journal-title":"J Biol Chem"},{"issue":"8","key":"pone.0353470.ref027","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1038\/nri1668","article-title":"Regulation of immune responses by L-arginine metabolism","volume":"5","author":"V Bronte","year":"2005","journal-title":"Nat Rev Immunol"},{"issue":"8","key":"pone.0353470.ref028","doi-asserted-by":"crossref","first-page":"3044","DOI":"10.1158\/0008-5472.CAN-04-4505","article-title":"Arginase-producing myeloid suppressor cells in renal cell carcinoma patients: a mechanism of tumor evasion","volume":"65","author":"AH Zea","year":"2005","journal-title":"Cancer Res"},{"issue":"1","key":"pone.0353470.ref029","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1152\/physrev.1995.75.1.77","article-title":"Experimental basis for neural-immune interactions","volume":"75","author":"KS Madden","year":"1995","journal-title":"Physiol Rev"},{"issue":"6","key":"pone.0353470.ref030","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1016\/j.it.2017.03.001","article-title":"Macrophage Immunometabolism: Where Are We (Going)?","volume":"38","author":"J Van den Bossche","year":"2017","journal-title":"Trends Immunol"},{"issue":"2","key":"pone.0353470.ref031","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1038\/nn.4477","article-title":"Neural regulation of immunity: molecular mechanisms and clinical translation","volume":"20","author":"VA Pavlov","year":"2017","journal-title":"Nat Neurosci"},{"issue":"4","key":"pone.0353470.ref032","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1016\/j.bbi.2009.10.015","article-title":"The immunoregulatory role of dopamine: an update","volume":"24","author":"C Sarkar","year":"2010","journal-title":"Brain Behav Immun"},{"key":"pone.0353470.ref033","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/j.anireprosci.2004.05.005","article-title":"Canine brucellosis","author":"MM Wanke","year":"2004","journal-title":"Anim Reprod Sci"},{"issue":"1","key":"pone.0353470.ref034","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1186\/1751-0147-54-18","article-title":"The first case of Brucella canis in Sweden: background, case report and recommendations from a northern European perspective","volume":"54","author":"BS Holst","year":"2012","journal-title":"Acta Vet Scand"},{"issue":"6084","key":"pone.0353470.ref035","doi-asserted-by":"crossref","first-page":"990","DOI":"10.1126\/science.1223066","article-title":"Cancer. Systems biology, metabolomics, and cancer metabolism","volume":"336","author":"M Tomita","year":"2012","journal-title":"Science"},{"issue":"9","key":"pone.0353470.ref036","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1038\/nri.2016.70","article-title":"A guide to immunometabolism for immunologists","volume":"16","author":"LAJ O\u2019Neill","year":"2016","journal-title":"Nat Rev Immunol"},{"issue":"1","key":"pone.0353470.ref037","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1038\/s41467-019-13668-3","article-title":"Mitochondrial TCA cycle metabolites control physiology and disease","volume":"11","author":"I Mart\u00ednez-Reyes","year":"2020","journal-title":"Nat Commun"},{"issue":"6","key":"pone.0353470.ref038","doi-asserted-by":"crossref","first-page":"1000","DOI":"10.1089\/ars.2013.5447","article-title":"Are reactive oxygen species always detrimental to pathogens?","volume":"20","author":"CN Paiva","year":"2014","journal-title":"Antioxid Redox Signal"},{"key":"pone.0353470.ref039","doi-asserted-by":"crossref","first-page":"929","DOI":"10.1146\/annurev.biochem.77.032207.120833","article-title":"Collagen structure and stability","volume":"78","author":"MD Shoulders","year":"2009","journal-title":"Annu Rev Biochem"},{"issue":"11","key":"pone.0353470.ref040","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1111\/j.1753-4887.2009.00246.x","article-title":"Choline: an essential nutrient for public health","volume":"67","author":"SH Zeisel","year":"2009","journal-title":"Nutr Rev"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0353470","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T17:48:50Z","timestamp":1784224130000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0353470"}},"subtitle":[],"editor":[{"given":"Roy Martin","family":"Roop II","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,7,16]]},"references-count":40,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7,16]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0353470","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,16]]}}}