{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T02:18:16Z","timestamp":1782094696231,"version":"3.54.5"},"reference-count":40,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,5,29]],"date-time":"2019-05-29T00:00:00Z","timestamp":1559088000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2019,5,29]],"date-time":"2019-05-29T00:00:00Z","timestamp":1559088000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100004022","name":"Jenny ja Antti Wihurin Rahasto","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004022","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Lyme borreliosis (LB), caused by bacteria of the <jats:italic>Borrelia burgdorferi<\/jats:italic> sensu lato (<jats:italic>Borrelia)<\/jats:italic> species, is the most common tick-borne infection in the northern hemisphere. LB diagnostics is based on clinical evaluation of the patient and on laboratory testing, where the main method is the detection of <jats:italic>Borrelia<\/jats:italic> specific antibodies in patient samples. There are, however, shortcomings in the current serology based LB diagnostics, especially its inability to differentiate ongoing infection from a previously treated one. Identification of specific biomarkers of diseases is a growing application of metabolomics. One of the main methods of metabolomics is nuclear magnetic resonance (NMR) spectroscopy. In the present study, our aim was to analyze whether <jats:italic>Borrelia<\/jats:italic> growth <jats:italic>in vitro<\/jats:italic> and infection <jats:italic>in vivo<\/jats:italic> in mice causes specific metabolite differences, and whether NMR can be used to detect them. For this purpose, we performed NMR analyses of <jats:italic>in vitro<\/jats:italic> culture medium samples, and of serum and urine samples of <jats:italic>Borrelia<\/jats:italic> infected and control mice. The results show, that there were significant differences in the concentrations of several amino acids, energy metabolites and aromatic compounds between <jats:italic>Borrelia<\/jats:italic> culture and control media, and between infected and control mouse serum and urine samples. For example, the concentration of <jats:sc>L<\/jats:sc>-phenylalanine increases in the <jats:italic>Borrelia<\/jats:italic> growth medium and in serum of infected mice, whereas the concentrations of allantoin and trigonelline decrease in the urine of infected mice. Therefore, we conclude that <jats:italic>Borrelia<\/jats:italic> infection causes measurable metabolome differences <jats:italic>in vitro<\/jats:italic> and in <jats:italic>Borrelia<\/jats:italic> infected mouse serum and urine samples, and that these can be detected with NMR.<\/jats:p>","DOI":"10.1038\/s41598-019-44540-5","type":"journal-article","created":{"date-parts":[[2019,5,29]],"date-time":"2019-05-29T10:02:24Z","timestamp":1559124144000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["NMR metabolome of Borrelia burgdorferi in vitro and in vivo in mice"],"prefix":"10.1038","volume":"9","author":[{"given":"Otto","family":"Glader","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elina","family":"Puljula","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johanna","family":"Jokioja","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maarit","family":"Karonen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jari","family":"Sinkkonen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jukka","family":"Hyt\u00f6nen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,5,29]]},"reference":[{"key":"44540_CR1","doi-asserted-by":"publisher","first-page":"461","DOI":"10.1016\/S0140-6736(11)60103-7","volume":"379","author":"G Stanek","year":"2012","unstructured":"Stanek, G., Wormser, G. P., Gray, J. & Strle, F. Lyme borreliosis. Lancet 379, 461\u2013473, https:\/\/doi.org\/10.1016\/S0140-6736(11)60103-7 (2012).","journal-title":"Lancet"},{"key":"44540_CR2","doi-asserted-by":"publisher","first-page":"1282","DOI":"10.3201\/eid2308.161273","volume":"23","author":"E Sajanti","year":"2017","unstructured":"Sajanti, E. et al. Lyme Borreliosis in Finland, 1995-2014. Emerg Infect Dis 23, 1282\u20131288, https:\/\/doi.org\/10.3201\/eid2308.161273 (2017).","journal-title":"Emerg Infect Dis"},{"key":"44540_CR3","doi-asserted-by":"publisher","first-page":"275","DOI":"10.1016\/j.ttbdis.2017.10.018","volume":"9","author":"J van Beek","year":"2018","unstructured":"van Beek, J. et al. Population-based Borrelia burgdorferi sensu lato seroprevalence and associated risk factors in Finland. Ticks Tick Borne Dis 9, 275\u2013280, https:\/\/doi.org\/10.1016\/j.ttbdis.2017.10.018 (2018).","journal-title":"Ticks Tick Borne Dis"},{"key":"44540_CR4","doi-asserted-by":"publisher","first-page":"484","DOI":"10.1128\/CMR.18.3.484-509.2005","volume":"18","author":"ME Aguero-Rosenfeld","year":"2005","unstructured":"Aguero-Rosenfeld, M. E., Wang, G., Schwartz, I. & Wormser, G. P. Diagnosis of lyme borreliosis. Clin Microbiol Rev 18, 484\u2013509, https:\/\/doi.org\/10.1128\/CMR.18.3.484-509.2005 (2005).","journal-title":"Clin Microbiol Rev"},{"key":"44540_CR5","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1111\/j.1574-695X.2006.00139.x","volume":"49","author":"B Wilske","year":"2007","unstructured":"Wilske, B., Fingerle, V. & Schulte-Spechtel, U. Microbiological and serological diagnosis of Lyme borreliosis. FEMS Immunol Med Microbiol 49, 13\u201321, https:\/\/doi.org\/10.1111\/j.1574-695X.2006.00139.x (2007).","journal-title":"FEMS Immunol Med Microbiol"},{"key":"44540_CR6","doi-asserted-by":"publisher","first-page":"473","DOI":"10.1038\/nrd.2016.32","volume":"15","author":"DS Wishart","year":"2016","unstructured":"Wishart, D. S. Emerging applications of metabolomics in drug discovery and precision medicine. Nat Rev Drug Discov 15, 473\u2013484, https:\/\/doi.org\/10.1038\/nrd.2016.32 (2016).","journal-title":"Nat Rev Drug Discov"},{"issue":"6","key":"44540_CR7","doi-asserted-by":"publisher","first-page":"354","DOI":"10.1038\/s41584-018-0005-9","volume":"14","author":"Virginia B. Kraus","year":"2018","unstructured":"Kraus, V. B. Biomarkers as drug development tools: discovery, validation, qualification and use. Nat Rev Rheumatol, https:\/\/doi.org\/10.1038\/s41584-018-0005-9 (2018).","journal-title":"Nature Reviews Rheumatology"},{"key":"44540_CR8","doi-asserted-by":"publisher","first-page":"730","DOI":"10.1097\/RCT.0000000000000424","volume":"40","author":"SH Choi","year":"2016","unstructured":"Choi, S. H., Kim, C. K., Park, J. J. & Park, B. K. Assessment of Early Therapeutic Changes to Concurrent Chemoradiotherapy in Uterine Cervical Cancer Using Blood Oxygenation Level-Dependent Magnetic Resonance Imaging. J Comput Assist Tomogr 40, 730\u2013734, https:\/\/doi.org\/10.1097\/RCT.0000000000000424 (2016).","journal-title":"J Comput Assist Tomogr"},{"key":"44540_CR9","doi-asserted-by":"publisher","first-page":"693","DOI":"10.1002\/mrc.4573","volume":"55","author":"S Kalantari","year":"2017","unstructured":"Kalantari, S., Nafar, M., Samavat, S. & Parvin, M. 1 H NMR-based metabolomics study for identifying urinary biomarkers and perturbed metabolic pathways associated with severity of IgA nephropathy: a pilot study. Magn Reson Chem 55, 693\u2013699, https:\/\/doi.org\/10.1002\/mrc.4573 (2017).","journal-title":"Magn Reson Chem"},{"key":"44540_CR10","doi-asserted-by":"publisher","DOI":"10.1038\/srep30785","volume":"6","author":"X Liu","year":"2016","unstructured":"Liu, X. et al. Identification of metabolic biomarkers in patients with type 2 diabetic coronary heart diseases based on metabolomic approach. Sci Rep 6, 30785, https:\/\/doi.org\/10.1038\/srep30785 (2016).","journal-title":"Sci Rep"},{"key":"44540_CR11","doi-asserted-by":"publisher","first-page":"3750","DOI":"10.1128\/JCM.01568-15","volume":"53","author":"SK Lau","year":"2015","unstructured":"Lau, S. K. et al. Metabolomic Profiling of Plasma from Patients with Tuberculosis by Use of Untargeted Mass Spectrometry Reveals Novel Biomarkers for Diagnosis. J Clin Microbiol 53, 3750\u20133759, https:\/\/doi.org\/10.1128\/JCM.01568-15 (2015).","journal-title":"J Clin Microbiol"},{"key":"44540_CR12","doi-asserted-by":"publisher","first-page":"e0006215","DOI":"10.1371\/journal.pntd.0006215","volume":"12","author":"E N\u00e4sstr\u00f6m","year":"2018","unstructured":"N\u00e4sstr\u00f6m, E. et al. Diagnostic metabolite biomarkers of chronic typhoid carriage. PLoS Negl Trop Dis 12, e0006215, https:\/\/doi.org\/10.1371\/journal.pntd.0006215 (2018).","journal-title":"PLoS Negl Trop Dis"},{"key":"44540_CR13","doi-asserted-by":"publisher","first-page":"e0004449","DOI":"10.1371\/journal.pntd.0004449","volume":"10","author":"NV Voge","year":"2016","unstructured":"Voge, N. V. et al. Metabolomics-Based Discovery of Small Molecule Biomarkers in Serum Associated with Dengue Virus Infections and Disease Outcomes. PLoS Negl Trop Dis 10, e0004449, https:\/\/doi.org\/10.1371\/journal.pntd.0004449 (2016).","journal-title":"PLoS Negl Trop Dis"},{"key":"44540_CR14","doi-asserted-by":"publisher","DOI":"10.1038\/srep02769","volume":"3","author":"L Tritten","year":"2013","unstructured":"Tritten, L. et al. Metabolic profiling framework for discovery of candidate diagnostic markers of malaria. Sci Rep 3, 2769, https:\/\/doi.org\/10.1038\/srep02769 (2013).","journal-title":"Sci Rep"},{"key":"44540_CR15","doi-asserted-by":"publisher","first-page":"1767","DOI":"10.1093\/cid\/civ185","volume":"60","author":"CR Molins","year":"2015","unstructured":"Molins, C. R. et al. Development of a metabolic biosignature for detection of early Lyme disease. Clin Infect Dis 60, 1767\u20131775, https:\/\/doi.org\/10.1093\/cid\/civ185 (2015).","journal-title":"Clin Infect Dis"},{"issue":"403","key":"44540_CR16","doi-asserted-by":"publisher","first-page":"eaal2717","DOI":"10.1126\/scitranslmed.aal2717","volume":"9","author":"Claudia R. Molins","year":"2017","unstructured":"Molins, C. R. et al. Metabolic differentiation of early Lyme disease from southern tick-associated rash illness (STARI). Sci Transl Med 9, https:\/\/doi.org\/10.1126\/scitranslmed.aal2717 (2017).","journal-title":"Science Translational Medicine"},{"key":"44540_CR17","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-29713-y","volume":"8","author":"A Pegalajar-Jurado","year":"2018","unstructured":"Pegalajar-Jurado, A. et al. Identification of Urine Metabolites as Biomarkers of Early Lyme Disease. Sci Rep 8, 12204, https:\/\/doi.org\/10.1038\/s41598-018-29713-y (2018).","journal-title":"Sci Rep"},{"key":"44540_CR18","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1016\/j.copbio.2016.08.001","volume":"43","author":"JL Markley","year":"2017","unstructured":"Markley, J. L. et al. The future of NMR-based metabolomics. Curr Opin Biotechnol 43, 34\u201340, https:\/\/doi.org\/10.1016\/j.copbio.2016.08.001 (2017).","journal-title":"Curr Opin Biotechnol"},{"key":"44540_CR19","doi-asserted-by":"publisher","DOI":"10.1186\/s12936-016-1256-z","volume":"15","author":"S Ghosh","year":"2016","unstructured":"Ghosh, S., Sengupta, A., Sharma, S. & Sonawat, H. M. Early prediction of cerebral malaria by (1)H NMR based metabolomics. Malar J 15, 198, https:\/\/doi.org\/10.1186\/s12936-016-1256-z (2016).","journal-title":"Malar J"},{"issue":"8","key":"44540_CR20","doi-asserted-by":"publisher","first-page":"e807","DOI":"10.1371\/journal.pntd.0000807","volume":"4","author":"Junfang Wu","year":"2010","unstructured":"Wu, J. et al. Metabolic changes reveal the development of schistosomiasis in mice. PLoS Negl Trop Dis 4, https:\/\/doi.org\/10.1371\/journal.pntd.0000807 (2010).","journal-title":"PLoS Neglected Tropical Diseases"},{"key":"44540_CR21","unstructured":"W, C. T. D. High-resolution NMR techniques in organic chemistry. Tetrahedron organic chemistry series 27 (2009)."},{"key":"44540_CR22","doi-asserted-by":"publisher","first-page":"1463","DOI":"10.1128\/IAI.72.3.1463-1469.2004","volume":"72","author":"WR Z\u00fcckert","year":"2004","unstructured":"Z\u00fcckert, W. R., Lloyd, J. E., Stewart, P. E., Rosa, P. A. & Barbour, A. G. Cross-species surface display of functional spirochetal lipoproteins by recombinant Borrelia burgdorferi. Infect Immun 72, 1463\u20131469 (2004).","journal-title":"Infect Immun"},{"key":"44540_CR23","doi-asserted-by":"publisher","first-page":"641","DOI":"10.1099\/0022-1317-51-8-641","volume":"51","author":"T Heikkil\u00e4","year":"2002","unstructured":"Heikkil\u00e4, T. et al. Cloning of the gene encoding the decorin-binding protein B (DbpB) in Borrelia burgdorferi sensu lato and characterisation of the antibody responses to DbpB in Lyme borreliosis. J Med Microbiol 51, 641\u2013648, https:\/\/doi.org\/10.1099\/0022-1317-51-8-641 (2002).","journal-title":"J Med Microbiol"},{"key":"44540_CR24","doi-asserted-by":"crossref","first-page":"2693","DOI":"10.1128\/jcm.28.12.2693-2699.1990","volume":"28","author":"JF Anderson","year":"1990","unstructured":"Anderson, J. F., Barthold, S. W. & Magnarelli, L. A. Infectious but nonpathogenic isolate of Borrelia burgdorferi. J Clin Microbiol 28, 2693\u20132699 (1990).","journal-title":"J Clin Microbiol"},{"key":"44540_CR25","doi-asserted-by":"publisher","first-page":"997","DOI":"10.1093\/ajcn\/29.9.997","volume":"29","author":"RW Wannemacher","year":"1976","unstructured":"Wannemacher, R. W., Klainer, A. S., Dinterman, R. E. & Beisel, W. R. The significance and mechanism of an increased serum phenylalanine-tyrosine ratio during infection. Am J Clin Nutr 29, 997\u20131006 (1976).","journal-title":"Am J Clin Nutr"},{"key":"44540_CR26","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1017\/S000711450769936X","volume":"98","author":"P Li","year":"2007","unstructured":"Li, P., Yin, Y. L., Li, D., Kim, S. W. & Wu, G. Amino acids and immune function. Br J Nutr 98, 237\u2013252, https:\/\/doi.org\/10.1017\/S000711450769936X (2007).","journal-title":"Br J Nutr"},{"key":"44540_CR27","doi-asserted-by":"publisher","first-page":"561","DOI":"10.1002\/bit.260380602","volume":"38","author":"D Duval","year":"1991","unstructured":"Duval, D., Demangel, C., Munier-Jolain, K., Miossec, S. & Geahel, I. Factors controlling cell proliferation and antibody production in mouse hybridoma cells: I. Influence of the amino acid supply. Biotechnol Bioeng 38, 561\u2013570, https:\/\/doi.org\/10.1002\/bit.260380602 (1991).","journal-title":"Biotechnol Bioeng"},{"key":"44540_CR28","doi-asserted-by":"publisher","first-page":"5433","DOI":"10.1021\/ac5005103","volume":"86","author":"GA Gowda","year":"2014","unstructured":"Gowda, G. A. & Raftery, D. Quantitating metabolites in protein precipitated serum using NMR spectroscopy. Anal Chem 86, 5433\u20135440, https:\/\/doi.org\/10.1021\/ac5005103 (2014). Nagana.","journal-title":"Anal Chem"},{"key":"44540_CR29","doi-asserted-by":"publisher","first-page":"e73076","DOI":"10.1371\/journal.pone.0073076","volume":"8","author":"S Bouatra","year":"2013","unstructured":"Bouatra, S. et al. The human urine metabolome. PLoS One 8, e73076, https:\/\/doi.org\/10.1371\/journal.pone.0073076 (2013).","journal-title":"PLoS One"},{"key":"44540_CR30","doi-asserted-by":"publisher","first-page":"665","DOI":"10.1111\/j.1600-0463.2010.02615.x","volume":"118","author":"H Yrj\u00e4n\u00e4inen","year":"2010","unstructured":"Yrj\u00e4n\u00e4inen, H., Hyt\u00f6nen, J., Hartiala, P., Oksi, J. & Viljanen, M. K. Persistence of borrelial DNA in the joints of Borrelia burgdorferi-infected mice after ceftriaxone treatment. APMIS 118, 665\u2013673, https:\/\/doi.org\/10.1111\/j.1600-0463.2010.02615.x (2010).","journal-title":"APMIS"},{"key":"44540_CR31","doi-asserted-by":"publisher","first-page":"e0121512","DOI":"10.1371\/journal.pone.0121512","volume":"10","author":"J Salo","year":"2015","unstructured":"Salo, J., Jaatinen, A., S\u00f6derstr\u00f6m, M., Viljanen, M. K. & Hyt\u00f6nen, J. Decorin Binding Proteins of Borrelia burgdorferi Promote Arthritis Development and Joint Specific Post-Treatment DNA Persistence in Mice. PLoS One 10, e0121512, https:\/\/doi.org\/10.1371\/journal.pone.0121512 (2015).","journal-title":"PLoS One"},{"key":"44540_CR32","doi-asserted-by":"publisher","first-page":"982","DOI":"10.1016\/j.foodchem.2012.10.081","volume":"138","author":"HM Lehtonen","year":"2013","unstructured":"Lehtonen, H. M. et al. 1H NMR-based metabolic fingerprinting of urine metabolites after consumption of lingonberries (Vaccinium vitis-idaea) with a high-fat meal. Food Chem 138, 982\u2013990, https:\/\/doi.org\/10.1016\/j.foodchem.2012.10.081 (2013).","journal-title":"Food Chem"},{"key":"44540_CR33","doi-asserted-by":"publisher","first-page":"190","DOI":"10.1016\/j.jmr.2009.11.012","volume":"202","author":"F Savorani","year":"2010","unstructured":"Savorani, F., Tomasi, G. & Engelsen, S. B. icoshift: A versatile tool for the rapid alignment of 1D NMR spectra. J Magn Reson 202, 190\u2013202, https:\/\/doi.org\/10.1016\/j.jmr.2009.11.012 (2010).","journal-title":"J Magn Reson"},{"key":"44540_CR34","doi-asserted-by":"publisher","first-page":"e201301009","DOI":"10.5936\/csbj.201301009","volume":"4","author":"J Bartel","year":"2013","unstructured":"Bartel, J., Krumsiek, J. & Theis, F. J. Statistical methods for the analysis of high-throughput metabolomics data. Comput Struct Biotechnol J 4, e201301009, https:\/\/doi.org\/10.5936\/csbj.201301009 (2013).","journal-title":"Comput Struct Biotechnol J"},{"key":"44540_CR35","doi-asserted-by":"publisher","first-page":"92","DOI":"10.2174\/2213235X11301010092","volume":"1","author":"B Worley","year":"2013","unstructured":"Worley, B. & Powers, R. Multivariate Analysis in Metabolomics. Curr Metabolomics 1, 92\u2013107, https:\/\/doi.org\/10.2174\/2213235X11301010092 (2013).","journal-title":"Curr Metabolomics"},{"key":"44540_CR36","unstructured":"Eriksson, L., Byrne, T., Johansson, E., Trygg, J. & Vikstr\u00f6m, C. Multi- and Megavariate Data Analysis: Basic principles and Applications. 3RD edn, 39\u201362 (MKS Umetrics AB, 2013)."},{"key":"44540_CR37","doi-asserted-by":"publisher","first-page":"D608","DOI":"10.1093\/nar\/gkx1089","volume":"46","author":"DS Wishart","year":"2018","unstructured":"Wishart, D. S. et al. HMDB 4.0: the human metabolome database for 2018. Nucleic Acids Res 46, D608\u2013D617, https:\/\/doi.org\/10.1093\/nar\/gkx1089 (2018).","journal-title":"Nucleic Acids Res"},{"key":"44540_CR38","doi-asserted-by":"publisher","first-page":"D603","DOI":"10.1093\/nar\/gkn810","volume":"37","author":"DS Wishart","year":"2009","unstructured":"Wishart, D. S. et al. HMDB: a knowledgebase for the human metabolome. Nucleic Acids Res 37, D603\u2013610, https:\/\/doi.org\/10.1093\/nar\/gkn810 (2009).","journal-title":"Nucleic Acids Res"},{"key":"44540_CR39","doi-asserted-by":"publisher","first-page":"D521","DOI":"10.1093\/nar\/gkl923","volume":"35","author":"DS Wishart","year":"2007","unstructured":"Wishart, D. S. et al. HMDB: the Human Metabolome Database. Nucleic Acids Res 35, D521\u2013526, https:\/\/doi.org\/10.1093\/nar\/gkl923 (2007).","journal-title":"Nucleic Acids Res"},{"key":"44540_CR40","doi-asserted-by":"publisher","first-page":"e16957","DOI":"10.1371\/journal.pone.0016957","volume":"6","author":"N Psychogios","year":"2011","unstructured":"Psychogios, N. et al. The human serum metabolome. PLoS One 6, e16957, https:\/\/doi.org\/10.1371\/journal.pone.0016957 (2011).","journal-title":"PLoS One"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-44540-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-44540-5","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-44540-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,17]],"date-time":"2022-12-17T10:08:53Z","timestamp":1671271733000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-44540-5"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,5,29]]},"references-count":40,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["44540"],"URL":"https:\/\/doi.org\/10.1038\/s41598-019-44540-5","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,5,29]]},"assertion":[{"value":"21 November 2018","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"20 May 2019","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"29 May 2019","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"8049"}}