{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T08:52:48Z","timestamp":1777366368132,"version":"3.51.4"},"reference-count":35,"publisher":"Springer Science and Business Media LLC","issue":"3","license":[{"start":{"date-parts":[[2022,11,17]],"date-time":"2022-11-17T00:00:00Z","timestamp":1668643200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2022,11,17]],"date-time":"2022-11-17T00:00:00Z","timestamp":1668643200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Biometals"],"published-print":{"date-parts":[[2023,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Salivary stones (sialoliths) are calcified structures located in the ductal system of the major salivary glands. Their exact cause is not clear but in general they are characterized by concentric inorganic (hydroxyapatite) layers. The formation is a slow intermittent process which may result in enlargement of the sialolith causing obstruction of saliva secretion resulting in mealtime related pain and swelling of the affected salivary gland. Various studies reported the presence of organic material such as proteins and lipids in the core of sialoliths. In the present study the protein composition of twenty submandibular sialoliths was analyzed. It was found that proteins contributed on average 5% to the dry weight of submandibular stones whereby small salivary stones contained more extractable proteins than large salivary stones. Using a combination of SDS-PAGE gel electrophoresis and Western blotting, we identified \u03b1-amylase (in all stones; 100%), lysozyme (95%), lactoferrin (85%), secretory-IgA (75%), MUC7 (60%), complement C4 (60%) and C-reactive protein (35%). The presence, and the combinations, of lactoferrin, lysozyme, s-IgA and \u03b1-amylase in sialoliths was confirmed by ELISA. The gradually increasing size of a sialolith might provoke a local inflammatory response in the duct of the submandibular gland whereby the relatively low concentrations of lactoferrin and lysozyme may originate from neutrophils. The interaction of lactoferrin with s-IgA could contribute to the accumulation of lactoferrin in sialoliths. In summary, these results suggest a new pathophysiological role for lactoferrin, in the formation of sialoliths.<\/jats:p>","DOI":"10.1007\/s10534-022-00465-7","type":"journal-article","created":{"date-parts":[[2022,11,17]],"date-time":"2022-11-17T21:03:01Z","timestamp":1668718981000},"page":"657-665","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Lactoferrin and the development of salivary stones: a pilot study"],"prefix":"10.1007","volume":"36","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4606-6588","authenticated-orcid":false,"given":"Saskia","family":"Kraaij","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jan G. A. M.","family":"de Visscher","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruben C.","family":"Apperloo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kamran","family":"Nazmi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Floris J.","family":"Bikker","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henk S.","family":"Brand","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2022,11,17]]},"reference":[{"issue":"5","key":"465_CR1","first-page":"36","volume":"82","author":"VV Afanas'ev","year":"2003","unstructured":"Afanas\u2019ev VV, Tkalenko AF, Abdusalamov MR (2003) Sostav rotovo\u012d zhidkosti smeshanno\u012d sliuny v zavisimosti ot rezul\u2019tatov lecheniia patsientov so sliunnokamenno\u012d bolezn\u2019iu s pomoshch\u2019iu metoda sialolitotripsii (Analysis of salivary pool composition in patients with different results of sialolithiasis treatment by sialo-lithotripsy). Stomatologiia Mosk 82(5):36\u201338","journal-title":"Stomatologiia Mosk"},{"issue":"8","key":"465_CR2","doi-asserted-by":"publisher","first-page":"778","DOI":"10.2174\/1389203715666140919124530","volume":"15","author":"AH Albar","year":"2014","unstructured":"Albar AH, Almehdar HA, Uversky VN, Redwan EM (2014) Structural heterogeneity and multifunctionality of lactoferrin. Curr Protein Pept Sci 15(8):778\u2013797. https:\/\/doi.org\/10.2174\/1389203715666140919124530","journal-title":"Curr Protein Pept Sci"},{"issue":"1","key":"465_CR3","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1034\/j.1601-0825.2002.1o816.x","volume":"8","author":"AV Amerongen","year":"2002","unstructured":"Amerongen AV, Veerman EC (2002) Saliva\u2013the defender of the oral cavity. Oral Dis 8(1):12\u201322. https:\/\/doi.org\/10.1034\/j.1601-0825.2002.1o816.x","journal-title":"Oral Dis"},{"issue":"7","key":"465_CR4","doi-asserted-by":"publisher","first-page":"4369","DOI":"10.1007\/s00784-020-03750-2","volume":"25","author":"G Avishai","year":"2021","unstructured":"Avishai G, Ben-Zvi Y, Chaushu G, Rosenfeld E, Gillman L, Reiser V, Gilat H (2021a) The unique characteristics of sialolithiasis following drug-induced hyposalivation. Clin Oral Investig 25(7):4369\u20134376. https:\/\/doi.org\/10.1007\/s00784-020-03750-2","journal-title":"Clin Oral Investig"},{"key":"465_CR5","doi-asserted-by":"publisher","DOI":"10.3390\/biology10050414","author":"G Avishai","year":"2021","unstructured":"Avishai G, Rabinovich I, Gilat H, Chaushu G, Chaushu L (2021b) Surgical treatment of sialolithiasis leads to improvement in the complete blood count. Biology (basel). https:\/\/doi.org\/10.3390\/biology10050414","journal-title":"Biology (basel)"},{"issue":"2","key":"465_CR6","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1016\/s0003-9969(02)00171-1","volume":"48","author":"L Becerra","year":"2003","unstructured":"Becerra L, Soares RV, Bruno LS, Siqueira CC, Oppenheim FG, Offner GD, Troxler RF (2003) Patterns of secretion of mucins and non-mucin glycoproteins in human submandibular\/sublingual secretion. Arch Oral Biol 48(2):147\u2013154. https:\/\/doi.org\/10.1016\/s0003-9969(02)00171-1","journal-title":"Arch Oral Biol"},{"issue":"7","key":"465_CR7","doi-asserted-by":"publisher","first-page":"1362","DOI":"10.1177\/00220345990780071101","volume":"78","author":"JGM Bolscher","year":"1999","unstructured":"Bolscher JGM, Groenink J, Van der Kwaak JS, Van den Keijbus PAM, Van\u2019t Hof W, Veerman ECI, NieuwAmerongen AV (1999) Detection and quantification of MUC7 in submandibular, sublingual, palatine, and labial saliva by anti-peptide antiserum. J Dent Res 78(7):1362\u20131369. https:\/\/doi.org\/10.1177\/00220345990780071101","journal-title":"J Dent Res"},{"key":"465_CR8","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1186\/s12014-020-09275-w","volume":"17","author":"CS Busso","year":"2020","unstructured":"Busso CS, Guidry JJ, Gonzalez JJ, Zorba V, Son LS, Winsauer PJ, Walvekar RR (2020) A comprehensive analysis of sialolith proteins and the clinical implications. Clin Proteom 17:12. https:\/\/doi.org\/10.1186\/s12014-020-09275-w","journal-title":"Clin Proteom"},{"issue":"4","key":"465_CR9","first-page":"161","volume":"27","author":"P Capaccio","year":"2007","unstructured":"Capaccio P, Torretta S, Ottavian F, Sambataro G, Pignataro L (2007) Modern management of obstructive salivary diseases. Acta Otorhinolaryngol Ital 27(4):161\u2013172","journal-title":"Acta Otorhinolaryngol Ital"},{"key":"465_CR10","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1159\/000358794","volume":"24","author":"K Delli","year":"2014","unstructured":"Delli K, Spijkervet FK, Vissink A (2014) Salivary gland diseases: infections, sialolithiasis and mucoceles. Monogr Oral Sci 24:135\u2013148. https:\/\/doi.org\/10.1159\/000358794","journal-title":"Monogr Oral Sci"},{"issue":"4","key":"465_CR11","doi-asserted-by":"publisher","first-page":"4295","DOI":"10.3390\/ijms13044295","volume":"13","author":"TK F\u00e1bi\u00e1n","year":"2012","unstructured":"F\u00e1bi\u00e1n TK, Hermann P, Beck A, Fej\u00e9rdy P, F\u00e1bi\u00e1n G (2012) Salivary defense proteins: their network and role in innate and acquired oral immunity. Int J Mol Sci 13(4):4295\u20134320. https:\/\/doi.org\/10.3390\/ijms13044295","journal-title":"Int J Mol Sci"},{"issue":"2","key":"465_CR12","doi-asserted-by":"publisher","first-page":"361","DOI":"10.1016\/S0095-5108(18)30057-5","volume":"26","author":"RP Garofalo","year":"1999","unstructured":"Garofalo RP, Goldman AS (1999) Expression of functional immunomodulatory and anti-inflammatory factors in human milk. Clin Perinatol 26(2):361\u2013377","journal-title":"Clin Perinatol"},{"issue":"1\u20132","key":"465_CR13","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/s0009-8981(03)00227-4","volume":"334","author":"F Grases","year":"2003","unstructured":"Grases F, Santiago C, Simonet BM, Costa-Bauz\u00e1 A (2003) Sialolithiasis: mechanism of calculi formation and etiologic factors. Clin Chim Acta 334(1\u20132):131\u2013136. https:\/\/doi.org\/10.1016\/s0009-8981(03)00227-4","journal-title":"Clin Chim Acta"},{"issue":"5","key":"465_CR14","doi-asserted-by":"publisher","first-page":"481","DOI":"10.1288\/00005537-195905000-00001","volume":"69","author":"JA Harrill","year":"1959","unstructured":"Harrill JA, King JS Jr, Boyce WH (1959) Structure and composition of salivary calculi. Laryngoscope 69(5):481\u2013492. https:\/\/doi.org\/10.1288\/00005537-195905000-00001","journal-title":"Laryngoscope"},{"issue":"6","key":"465_CR33","doi-asserted-by":"publisher","first-page":"927","DOI":"10.1016\/j.otc.2009.08.012","volume":"42","author":"John D. Harrison","year":"2009","unstructured":"Harrison JD (2009) Causes natural history and incidence of salivary stones and obstructions. Otolaryngol Clin N Am 42(6):927\u2013947. https:\/\/doi.org\/10.1016\/j.otc.2009.08.012","journal-title":"Otolaryngologic Clinics of North America"},{"issue":"1","key":"465_CR15","doi-asserted-by":"publisher","first-page":"152","DOI":"10.1288\/00005537-198001000-00016","volume":"90","author":"F Hiraide","year":"1980","unstructured":"Hiraide F, Nomura Y (1980) The fine surface structure and composition of salivary calculi. Laryngoscope 90(1):152\u2013158. https:\/\/doi.org\/10.1288\/00005537-198001000-00016","journal-title":"Laryngoscope"},{"key":"465_CR35","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1155\/2015\/285237","volume":"2015","author":"Xiao-Min Hu","year":"2015","unstructured":"Hu XM, Xu YR, Yan R, Sun SL, Dong HL, Wang J, Gao XM (2015) A novel murine anti-lactoferrin monoclonal antibody activates human polymorphonuclear leukocytes through membrane-bound lactoferrin and TLR4. Biomed Res Int 1\u201311. https:\/\/doi.org\/10.1155\/2015\/285237","journal-title":"BioMed Research International"},{"issue":"3","key":"465_CR16","doi-asserted-by":"publisher","first-page":"217","DOI":"10.1111\/j.1600-0714.1983.tb00335.x","volume":"12","author":"G Isacsson","year":"1983","unstructured":"Isacsson G, Hammarstr\u00f6m L (1983) An enzyme histochemical study of human salivary duct calculi. J Oral Pathol 12(3):217\u2013222. https:\/\/doi.org\/10.1111\/j.1600-0714.1983.tb00335.x","journal-title":"J Oral Pathol"},{"issue":"10","key":"465_CR17","doi-asserted-by":"publisher","first-page":"1253","DOI":"10.1016\/j.joms.2003.11.018","volume":"62","author":"O Kasabo\u011flu","year":"2004","unstructured":"Kasabo\u011flu O, Er N, T\u00fcmer C, Akkocao\u011flu M (2004) Micromorphology of sialoliths in submandibular salivary gland: a scanning electron microscope and X-ray diffraction analysis. J Oral Maxillofac Surg 62(10):1253\u20131258. https:\/\/doi.org\/10.1016\/j.joms.2003.11.018","journal-title":"J Oral Maxillofac Surg"},{"issue":"11","key":"465_CR18","doi-asserted-by":"publisher","first-page":"E23","DOI":"10.1038\/sj.bdj.2014.1054","volume":"217","author":"S Kraaij","year":"2014","unstructured":"Kraaij S, Karagozoglu KH, Forouzanfar T, Veerman EC, Brand HS (2014) Salivary stones: symptoms, aetiology, biochemical composition and treatment. Br Dent J 217(11):E23. https:\/\/doi.org\/10.1038\/sj.bdj.2014.1054","journal-title":"Br Dent J"},{"issue":"1","key":"465_CR19","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1111\/j.1600-0722.1996.tb00040.x","volume":"104","author":"J Kugler","year":"1996","unstructured":"Kugler J, Breitfeld I, Tewes U, Schedlowski M (1996) Excavation of caries lesions induces transient decrease of total salivary immunoglobulin A concentration. Eur J Oral Sci 104(1):17\u201320. https:\/\/doi.org\/10.1111\/j.1600-0722.1996.tb00040.x","journal-title":"Eur J Oral Sci"},{"issue":"2","key":"465_CR32","doi-asserted-by":"publisher","first-page":"307","DOI":"10.1007\/s00270-009-9731-3","volume":"33","author":"I. Nagra","year":"2010","unstructured":"Nagra I, Jones C, Dyer J (2010) Endoluminal Intervention in the Salivary Duct: Clinical Outcomes at a District General Hospital. CardioVascular and Interventional Radiology 33(2):307\u2013314. https:\/\/doi.org\/10.1007\/s00270-009-9731-3","journal-title":"CardioVascular and Interventional Radiology"},{"issue":"5","key":"465_CR20","doi-asserted-by":"publisher","first-page":"1190","DOI":"10.1017\/s1431927613001694","volume":"19","author":"P Nolasco","year":"2013","unstructured":"Nolasco P, Anjos AJ, Marques JM, Cabrita F, Da Costa EC, Maur\u00edcio A, Pereira MF, De Matos AP, Carvalho PA (2013) Structure and growth of sialoliths: computed microtomography and electron microscopy investigation of 30 specimens. Microsc Microanal 19(5):1190\u20131203. https:\/\/doi.org\/10.1017\/s1431927613001694","journal-title":"Microsc Microanal"},{"issue":"3","key":"465_CR21","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1007\/bf02008226","volume":"16","author":"CI Osuoji","year":"1974","unstructured":"Osuoji CI, Rowles SL (1974) Studies on the organic composition of dental calculus and related calculi. Calcif Tissue Res 16(3):193\u2013200. https:\/\/doi.org\/10.1007\/bf02008226","journal-title":"Calcif Tissue Res"},{"key":"465_CR34","unstructured":"Proctor GB, McGurk M, Harrison JD  (2005) Protein composition of submandibular stones. J Dent Res 84 (special issue) A218 (congres abstract)"},{"issue":"3","key":"465_CR22","doi-asserted-by":"publisher","first-page":"149","DOI":"10.1111\/eos.12182","volume":"123","author":"A Prodan","year":"2015","unstructured":"Prodan A, Brand HS, Ligtenberg AJ, Imangaliyev S, Tsivtsivadze E, Van Der Weijden F, Crielaard W, Keijser BJ, Veerman EC (2015) Interindividual variation, correlations, and sex-related differences in the salivary biochemistry of young healthy adults. Eur J Oral Sci 123(3):149\u2013157. https:\/\/doi.org\/10.1111\/eos.12182","journal-title":"Eur J Oral Sci"},{"issue":"1","key":"465_CR23","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1139\/bcb-2020-0052","volume":"99","author":"L Rosa","year":"2021","unstructured":"Rosa L, Lepanto MS, Cutone A, Ianiro G, Pernarella S, Sangermano R, Musci G, Ottolenghi L, Valenti P (2021) Lactoferrin and oral pathologies: a therapeutic treatment. Biochem Cell Biol 99(1):81\u201390. https:\/\/doi.org\/10.1139\/bcb-2020-0052","journal-title":"Biochem Cell Biol"},{"issue":"4","key":"465_CR24","doi-asserted-by":"publisher","first-page":"526","DOI":"10.4103\/ccd.ccd_103_17","volume":"8","author":"A Sharma","year":"2017","unstructured":"Sharma A, Subramaniam P, Moiden S (2017) Analysis of salivary IgA, amylase, lactoferrin, and lysozyme before and after comprehensive dental treatment in children: a prospective study. Contemp Clin Dent 8(4):526\u2013530. https:\/\/doi.org\/10.4103\/ccd.ccd_103_17","journal-title":"Contemp Clin Dent"},{"issue":"8","key":"465_CR25","doi-asserted-by":"publisher","first-page":"866","DOI":"10.1177\/00220345830620080201","volume":"62","author":"BL Slomiany","year":"1983","unstructured":"Slomiany BL, Murty VL, Aono M, Slomiany A, Mandel ID (1983) Lipid composition of human parotid salivary gland stones. J Dent Res 62(8):866\u2013869. https:\/\/doi.org\/10.1177\/00220345830620080201","journal-title":"J Dent Res"},{"issue":"1","key":"465_CR26","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1016\/j.archoralbio.2009.11.006","volume":"55","author":"YX Su","year":"2010","unstructured":"Su YX, Zhang K, Ke ZF, Zheng GS, Chu M, Liao GQ (2010) Increased calcium and decreased magnesium and citrate concentrations of submandibular\/sublingual saliva in sialolithiasis. Arch Oral Biol 55(1):15\u201320. https:\/\/doi.org\/10.1016\/j.archoralbio.2009.11.006","journal-title":"Arch Oral Biol"},{"issue":"7","key":"465_CR27","doi-asserted-by":"publisher","first-page":"1709","DOI":"10.1007\/s00784-012-0870-6","volume":"17","author":"J Szalma","year":"2013","unstructured":"Szalma J, B\u00f6ddi K, Lempel E, Sieroslawska AF, Szab\u00f3 Z, Harfouche R, Olasz L, Tak\u00e1tsy A, Guttman A (2013) Proteomic and scanning electron microscopic analysis of submandibular sialoliths. Clin Oral Invest 17(7):1709\u20131717. https:\/\/doi.org\/10.1007\/s00784-012-0870-6","journal-title":"Clin Oral Invest"},{"issue":"3","key":"465_CR28","doi-asserted-by":"publisher","first-page":"247","DOI":"10.1159\/000077762","volume":"38","author":"AA Van Nieuw","year":"2004","unstructured":"Van Nieuw AA, Bolscher JG, Veerman EC (2004) Salivary proteins: protective and diagnostic value in cariology? Caries Res 38(3):247\u2013253. https:\/\/doi.org\/10.1159\/000077762","journal-title":"Caries Res"},{"issue":"22","key":"465_CR29","doi-asserted-by":"publisher","first-page":"2540","DOI":"10.1007\/s00018-005-5369-8","volume":"62","author":"PP Ward","year":"2005","unstructured":"Ward PP, Paz E, Conneely OM (2005) Multifunctional roles of lactoferrin: a critical overview. Cell Mol Life Sci 62(22):2540\u20132548. https:\/\/doi.org\/10.1007\/s00018-005-5369-8","journal-title":"Cell Mol Life Sci"},{"issue":"1","key":"465_CR30","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1016\/s0003-9969(96)00104-5","volume":"42","author":"CK Yeh","year":"1997","unstructured":"Yeh CK, Dodds MW, Zuo P, Johnson DA (1997) A population-based study of salivary lysozyme concentrations and candidal counts. Arch Oral Biol 42(1):25\u201331. https:\/\/doi.org\/10.1016\/s0003-9969(96)00104-5","journal-title":"Arch Oral Biol"},{"issue":"11","key":"465_CR31","doi-asserted-by":"publisher","first-page":"2243","DOI":"10.1007\/s10856-006-0057-2","volume":"18","author":"XD Zhu","year":"2007","unstructured":"Zhu XD, Fan HS, Zhao CY, Lu J, Ikoma T, Tanaka J, Zhang XD (2007) Competitive adsorption of bovine serum albumin and lysozyme on characterized calcium phosphates by polyacrylamide gel electrophoresis method. J Mater Sci Mater Med 18(11):2243\u20132249. https:\/\/doi.org\/10.1007\/s10856-006-0057-2","journal-title":"J Mater Sci Mater Med"}],"container-title":["BioMetals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s10534-022-00465-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s10534-022-00465-7\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s10534-022-00465-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,5,12]],"date-time":"2023-05-12T10:27:51Z","timestamp":1683887271000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s10534-022-00465-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,17]]},"references-count":35,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2023,6]]}},"alternative-id":["465"],"URL":"https:\/\/doi.org\/10.1007\/s10534-022-00465-7","relation":{},"ISSN":["0966-0844","1572-8773"],"issn-type":[{"value":"0966-0844","type":"print"},{"value":"1572-8773","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,11,17]]},"assertion":[{"value":"5 May 2022","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"5 November 2022","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"17 November 2022","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors have no competing interests to declare that are relevant to the content of this article.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}},{"value":"Our project \u2018Research on the composition of salivary stones\u2019 (in Dutch: Onderzoek naar de samenstelling van speekselstenen\u2019) has been approved by the Medical Ethical Committee of the Amsterdam UMC location VUmc (protocol number 2012\/127).","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical approval"}}]}}