{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,13]],"date-time":"2025-05-13T15:21:49Z","timestamp":1747149709461,"version":"3.40.5"},"reference-count":23,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2022,2,6]],"date-time":"2022-02-06T00:00:00Z","timestamp":1644105600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100009553","name":"Agentura Pro Zdravotnick\u00fd V\u00fdzkum \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["AZV CR NV16\u201033923A"],"award-info":[{"award-number":["AZV CR NV16\u201033923A"]}],"id":[{"id":"10.13039\/501100009553","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J of Inher Metab Disea"],"published-print":{"date-parts":[[2022,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Deuterium\u2010labeled cholesterol\u2010dextran particles (d4\u2010CholDex), prepared by co\u2010precipitation, were internalized by cultured human skin fibroblasts and HEK293 cells. Subcellular particles from d4\u2010CholDex\u2010treated HEK293 cells were fractionated on iodixanol gradients. More than 60% of d4\u2010cholesterol (d4\u2010UC) in the gradient co\u2010fractionated with lysosomal markers and NPC1. This and formation of d4\u2010cholesteryl esters (d4\u2010CE) in the cells suggests that d4\u2010CholDex is lysosomally processed. In accordance with these findings, we observed an increase in lysosomal cholesterol content by fluorescence microscopy in CholDex\u2010loaded cells. Fibroblast cultures including 13 NPC1\u2010deficient, four heterozygous and six control lines were treated with d4\u2010CholDex at final d4\u2010UC concentration of 0.05\u2009mg\/ml (127.98\u00a0\u03bcmol\/L) for 3\u00a0h and chased for 48\u2009h in medium without d4\u2010CholDex. Concentrations of d4\u2010UC and d4\u2010CE in harvested cells were measured by tandem mass spectrometry (MS\/MS). d4\u2010UC\/d4\u2010CE ratios were elevated in NP\u2010C lines compared to controls (<jats:italic>n<\/jats:italic>\u00a0=\u00a06, mean\u00a0=\u00a04.36, range\u00a0=\u00a01.89\u20138.91), with the highest ratios in severe NP\u2010C1 phenotypes and the lowest in adolescent\/adult type patients. There were overlaps between NP\u2010C1 forms: early infantile (<jats:italic>n<\/jats:italic>\u00a0=\u00a01, mean\u00a0=\u00a048.6), late infantile (<jats:italic>n<\/jats:italic>\u00a0=\u00a04, mean\u00a0=\u00a036.3, range\u00a0=\u00a020.6\u201354.0), juvenile (<jats:italic>n<\/jats:italic>\u00a0=\u00a05, mean\u00a0=\u00a024.7, range\u00a0=\u00a013.4\u201338.3), adolescent\/adult (<jats:italic>n<\/jats:italic>\u00a0=\u00a03, mean\u00a0=\u00a014.5, range\u00a0=\u00a011.7\u201319.8). The ratios in NP\u2010C1 heterozygotes were mildly elevated (<jats:italic>n<\/jats:italic>\u00a0=\u00a04, mean\u00a0=\u00a016.4, range\u00a0=\u00a014.9\u201317.4) and comparable to patients with adolescent\/adult NP\u2010C1. The test can be useful in evaluation of suspected NP\u2010C patients with inconclusive results of biomarker or molecular tests. Its advantages include standardized preparation of particles with longer shelf life at 4\u00a0\u00b0C, quantitative results, and no requirement for radioactive chemicals.<\/jats:p>","DOI":"10.1002\/jimd.12481","type":"journal-article","created":{"date-parts":[[2022,1,28]],"date-time":"2022-01-28T11:49:01Z","timestamp":1643370541000},"page":"584-592","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Loading of cell cultures with cholesterol\u2010dextran particles as a new functional test for <scp>Niemann\u2013Pick<\/scp> type C disease"],"prefix":"10.1002","volume":"45","author":[{"given":"Filip","family":"Majer","sequence":"first","affiliation":[{"name":"Research Unit for Rare Diseases, Department of Pediatrics and Inherited Metabolic Disorders, First Faculty of Medicine Charles University and General University Hospital  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Befekadu","family":"Asfaw","sequence":"additional","affiliation":[{"name":"Research Unit for Rare Diseases, Department of Pediatrics and Inherited Metabolic Disorders, First Faculty of Medicine Charles University and General University Hospital  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ladislav","family":"Kucha\u0159","sequence":"additional","affiliation":[{"name":"Research Unit for Rare Diseases, Department of Pediatrics and Inherited Metabolic Disorders, First Faculty of Medicine Charles University and General University Hospital  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dita","family":"Mu\u0161\u00e1lkov\u00e1","sequence":"additional","affiliation":[{"name":"Research Unit for Rare Diseases, Department of Pediatrics and Inherited Metabolic Disorders, First Faculty of Medicine Charles University and General University Hospital  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lenka","family":"Steiner\u2010Mr\u00e1zov\u00e1","sequence":"additional","affiliation":[{"name":"Research Unit for Rare Diseases, Department of Pediatrics and Inherited Metabolic Disorders, First Faculty of Medicine Charles University and General University Hospital  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert","family":"Dobrovoln\u00fd","sequence":"additional","affiliation":[{"name":"Research Unit for Rare Diseases, Department of Pediatrics and Inherited Metabolic Disorders, First Faculty of Medicine Charles University and General University Hospital  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jana","family":"Ledvinov\u00e1","sequence":"additional","affiliation":[{"name":"Research Unit for Rare Diseases, Department of Pediatrics and Inherited Metabolic Disorders, First Faculty of Medicine Charles University and General University Hospital  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0628-8917","authenticated-orcid":false,"given":"Martin","family":"H\u0159eb\u00ed\u010dek","sequence":"additional","affiliation":[{"name":"Research Unit for Rare Diseases, Department of Pediatrics and Inherited Metabolic Disorders, First Faculty of Medicine Charles University and General University Hospital  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2022,2,6]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1186\/1750\u20101172\u20105\u201016"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1022575511354"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010019\u201011425\u20100"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.21635"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/bs.mcb.2014.10.028"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1194\/jlr.P013524"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1399\u20100004.1988.tb03460.x"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2010.04.047"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13023\u2010018\u20100785\u20107"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgme.2016.06.004"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0014\u20105793(98)01578\u20106"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/978\u201094\u2010007\u20102555\u20103_11"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0008\u20106215(00)82357\u20105"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.3390\/molecules170911067"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003\u20102697(72)90094\u20102"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbalip.2005.12.007"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13023\u2010020\u201001360\u20105"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI16452"},{"key":"e_1_2_11_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2009.07.003"},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF02892554"},{"key":"e_1_2_11_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.atherosclerosis.2019.09.022"},{"key":"e_1_2_11_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature17664"},{"key":"e_1_2_11_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2014.06.016"}],"container-title":["Journal of Inherited Metabolic Disease"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jimd.12481","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/jimd.12481","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jimd.12481","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,23]],"date-time":"2023-08-23T08:02:35Z","timestamp":1692777755000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jimd.12481"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,6]]},"references-count":23,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2022,5]]}},"alternative-id":["10.1002\/jimd.12481"],"URL":"https:\/\/doi.org\/10.1002\/jimd.12481","archive":["Portico"],"relation":{},"ISSN":["0141-8955","1573-2665"],"issn-type":[{"type":"print","value":"0141-8955"},{"type":"electronic","value":"1573-2665"}],"subject":[],"published":{"date-parts":[[2022,2,6]]},"assertion":[{"value":"2021-10-14","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-01-25","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-02-06","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}