{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T01:02:20Z","timestamp":1759971740038,"version":"build-2065373602"},"reference-count":54,"publisher":"Wiley","issue":"7-8","license":[{"start":{"date-parts":[[2022,8,30]],"date-time":"2022-08-30T00:00:00Z","timestamp":1661817600000},"content-version":"vor","delay-in-days":29,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Analytical Science Advances"],"published-print":{"date-parts":[[2022,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p><jats:italic>Neospora caninum<\/jats:italic> is an obligate intracellular protozoan parasite of the phylum Alveolata (subphylum Apicomplexa) which has not been studied extensively in a biochemical context. <jats:italic>N. caninum<\/jats:italic> is a primary cause of reproductive disorders causing mummification and abortion not only in cattle but also in other small ruminant species resulting in a substantial economic impact on the livestock industry. In canids, which are the final hosts of <jats:italic>N. caninum<\/jats:italic>, clinical disease includes neuromuscular symptoms, ataxia, and ascending paralysis. Fatal outcomes of neosporosis have also been reported depending on the host species, age and immune status, however, its zoonotic potential is still uncertain. Therefore, <jats:italic>N. caninum<\/jats:italic> should be thoroughly investigated. Matrix\u2010assisted laser desorption\/ionisation (MALDI) mass spectrometry (MS) and MS imaging (MSI) were used, combined with high\u2010performance liquid chromatography (HPLC) to investigate these intracellular parasites. The aim of this study was to identify molecular biomarkers for <jats:italic>N. caninum<\/jats:italic> tachyzoite\u2010infected host cells and to further clarify their functions. By atmospheric\u2010pressure scanning microprobe MALDI MS(I), sections of <jats:italic>N. caninum<\/jats:italic>\u2010infected and non\u2010infected host cell pellets were examined in order to determine potential markers. In vivo, <jats:italic>N. caninum<\/jats:italic> infects different types of nucleated cells, such as endothelial cells which represent a highly immunoreactive cell type. Therefore, primary bovine umbilical vein endothelial cells were here used as a suitable infection system. For comparison, the permanent MARC\u2010145 cell line was used as an additional, simplified in vitro cell culture model. HPLC\u2010tandem MS (HPLC\u2010MS\/MS) experiments combined with database search were employed for structural verification of markers. The statistically relevant biomarkers found by MS and identified by HPLC\u2010MS\/MS measurements were partly also found in infected monolayers. Marker signals were imaged in cell layers of <jats:italic>N. caninum<\/jats:italic>\u2010infected and non\u2010infected host cells at 5\u00a0\u00b5m lateral resolution.<\/jats:p>","DOI":"10.1002\/ansa.202200016","type":"journal-article","created":{"date-parts":[[2022,8,30]],"date-time":"2022-08-30T13:04:46Z","timestamp":1661864686000},"page":"244-254","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Atmospheric\u2010pressure scanning microprobe matrix\u2010assisted laser desorption\/ionization mass spectrometry imaging of <i>Neospora caninum<\/i>\u2010infected cell monolayers"],"prefix":"10.1002","volume":"3","author":[{"given":"Nils H.","family":"Ansch\u00fctz","sequence":"first","affiliation":[{"name":"Institute of Inorganic and Analytical Chemistry Justus Liebig University Giessen  Giessen Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefanie","family":"Gerbig","sequence":"additional","affiliation":[{"name":"Institute of Inorganic and Analytical Chemistry Justus Liebig University Giessen  Giessen Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alejandra M. Peter","family":"Ventura","sequence":"additional","affiliation":[{"name":"Institute of Inorganic and Analytical Chemistry Justus Liebig University Giessen  Giessen Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Liliana M. R.","family":"Silva","sequence":"additional","affiliation":[{"name":"Institute of Parasitology Justus Liebig University Giessen  Giessen Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Camilo","family":"Larrazabal","sequence":"additional","affiliation":[{"name":"Institute of Parasitology Justus Liebig University Giessen  Giessen Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carlos","family":"Hermosilla","sequence":"additional","affiliation":[{"name":"Institute of Parasitology Justus Liebig University Giessen  Giessen Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anja","family":"Taubert","sequence":"additional","affiliation":[{"name":"Institute of Parasitology Justus Liebig University Giessen  Giessen Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bernhard","family":"Spengler","sequence":"additional","affiliation":[{"name":"Institute of Inorganic and Analytical Chemistry Justus Liebig University Giessen  Giessen Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2022,8,30]]},"reference":[{"doi-asserted-by":"publisher","key":"e_1_2_10_2_1","DOI":"10.1186\/s13071\u2010018\u20102683\u20101"},{"issue":"9","key":"e_1_2_10_3_1","doi-asserted-by":"crossref","first-page":"1269","DOI":"10.2460\/javma.1988.192.09.1269","article-title":"Newly recognized fatal protozoan disease of dogs","volume":"192","author":"Dubey JP","year":"1988","journal-title":"J Am Vet Med Assoc"},{"doi-asserted-by":"publisher","key":"e_1_2_10_4_1","DOI":"10.1016\/s0020\u20107519(99)00121\u20106"},{"doi-asserted-by":"publisher","key":"e_1_2_10_5_1","DOI":"10.1016\/s0020\u20107519(98)00138\u20106"},{"doi-asserted-by":"publisher","key":"e_1_2_10_6_1","DOI":"10.1016\/s0020\u20107519(99)00132\u20100"},{"doi-asserted-by":"publisher","key":"e_1_2_10_7_1","DOI":"10.1371\/journal.pone.0124473"},{"doi-asserted-by":"publisher","key":"e_1_2_10_8_1","DOI":"10.1371\/journal.ppat.1002567"},{"doi-asserted-by":"publisher","key":"e_1_2_10_9_1","DOI":"10.1016\/s0020\u20107519(02)00094\u20102"},{"doi-asserted-by":"publisher","key":"e_1_2_10_10_1","DOI":"10.1128\/cmr.00031\u201006"},{"doi-asserted-by":"publisher","key":"e_1_2_10_11_1","DOI":"10.1016\/j.ijpara.2012.10.022"},{"doi-asserted-by":"publisher","key":"e_1_2_10_12_1","DOI":"10.1007\/s004360050355"},{"doi-asserted-by":"publisher","key":"e_1_2_10_13_1","DOI":"10.1016\/s0304\u20104017(96)01076\u2010x"},{"doi-asserted-by":"publisher","key":"e_1_2_10_14_1","DOI":"10.1111\/j.1365\u20103024.1994.tb00320.x"},{"doi-asserted-by":"publisher","key":"e_1_2_10_15_1","DOI":"10.1016\/0020\u20107519(96)00030\u20106"},{"doi-asserted-by":"publisher","key":"e_1_2_10_16_1","DOI":"10.1016\/j.pt.2005.09.005"},{"doi-asserted-by":"publisher","key":"e_1_2_10_17_1","DOI":"10.1016\/s0020\u20107519(99)00129\u20100"},{"doi-asserted-by":"publisher","key":"e_1_2_10_18_1","DOI":"10.2147\/vmrr.s76969"},{"doi-asserted-by":"publisher","key":"e_1_2_10_19_1","DOI":"10.1007\/s11250\u2010017\u20101403\u20108"},{"doi-asserted-by":"publisher","key":"e_1_2_10_20_1","DOI":"10.1016\/j.micpath.2015.12.015"},{"doi-asserted-by":"publisher","key":"e_1_2_10_21_1","DOI":"10.1016\/s0304\u20104017(96)01035\u20107"},{"doi-asserted-by":"publisher","key":"e_1_2_10_22_1","DOI":"10.1016\/j.vetpar.2016.01.006"},{"doi-asserted-by":"publisher","key":"e_1_2_10_23_1","DOI":"10.1016\/j.prevetmed.2011.08.010"},{"issue":"2","key":"e_1_2_10_24_1","first-page":"236","article-title":"Experimental fetal and transplacental neospora infection in the nonhuman primate","volume":"71","author":"Barr BC","year":"1994","journal-title":"Lab Invest"},{"doi-asserted-by":"publisher","key":"e_1_2_10_25_1","DOI":"10.21897\/rmvz.1139"},{"doi-asserted-by":"publisher","key":"e_1_2_10_26_1","DOI":"10.1038\/s41598\u2010020\u201065991\u20101"},{"doi-asserted-by":"publisher","key":"e_1_2_10_27_1","DOI":"10.1590\/0037\u20108682\u20100151\u20102015"},{"doi-asserted-by":"publisher","key":"e_1_2_10_28_1","DOI":"10.1007\/s00436\u2010017\u20105579\u20107"},{"doi-asserted-by":"publisher","key":"e_1_2_10_29_1","DOI":"10.1016\/j.jasms.2004.01.007"},{"doi-asserted-by":"publisher","key":"e_1_2_10_30_1","DOI":"10.1255\/ejms.840"},{"doi-asserted-by":"publisher","key":"e_1_2_10_31_1","DOI":"10.1016\/j.jasms.2008.08.005"},{"doi-asserted-by":"publisher","key":"e_1_2_10_32_1","DOI":"10.1038\/nmeth.4071"},{"unstructured":"SpenglerB HubertM KaufmannR.MALDI ion imaging and biological ion imaging with a new scanning UV\u2010laser microprobe. InProceedings of the 42nd ASMS Conference on Mass Spectrometry and Allied Topics.1994.","key":"e_1_2_10_33_1"},{"doi-asserted-by":"publisher","key":"e_1_2_10_34_1","DOI":"10.1021\/jasms.0c00043"},{"doi-asserted-by":"publisher","key":"e_1_2_10_35_1","DOI":"10.1016\/j.ijpara.2006.01.001"},{"doi-asserted-by":"publisher","key":"e_1_2_10_36_1","DOI":"10.1194\/jlr.D700041\u2010JLR200"},{"doi-asserted-by":"publisher","key":"e_1_2_10_37_1","DOI":"10.1021\/pr800373m"},{"doi-asserted-by":"publisher","key":"e_1_2_10_38_1","DOI":"10.1038\/nmeth.3901"},{"doi-asserted-by":"publisher","key":"e_1_2_10_39_1","DOI":"10.1186\/s12859\u2010017\u20101744\u20103"},{"doi-asserted-by":"publisher","key":"e_1_2_10_40_1","DOI":"10.1093\/bioinformatics\/btn323"},{"doi-asserted-by":"publisher","key":"e_1_2_10_41_1","DOI":"10.1186\/1471\u20102105\u201011\u2010395"},{"doi-asserted-by":"publisher","key":"e_1_2_10_42_1","DOI":"10.4161\/spmg.19885"},{"doi-asserted-by":"publisher","key":"e_1_2_10_43_1","DOI":"10.1051\/vetres\/2010041"},{"doi-asserted-by":"publisher","key":"e_1_2_10_44_1","DOI":"10.1016\/j.vetinn.2006.03.017"},{"doi-asserted-by":"publisher","key":"e_1_2_10_45_1","DOI":"10.1016\/j.ijmm.2012.07.002"},{"doi-asserted-by":"publisher","key":"e_1_2_10_46_1","DOI":"10.1007\/s00436\u2010001\u20100531\u20101"},{"unstructured":"The LIPID MAPS\u00aeLipidomics Gateway.2020.https:\/\/www.lipidmaps.org\/","key":"e_1_2_10_47_1"},{"doi-asserted-by":"publisher","key":"e_1_2_10_48_1","DOI":"10.1002\/(sici)1097-0231(20000331)14:6<390::aid-rcm882>3.0.co;2-7"},{"doi-asserted-by":"publisher","key":"e_1_2_10_49_1","DOI":"10.1002\/rcm.2820"},{"doi-asserted-by":"publisher","key":"e_1_2_10_50_1","DOI":"10.1194\/jlr.D800040\u2010JLR200"},{"doi-asserted-by":"publisher","key":"e_1_2_10_51_1","DOI":"10.1194\/jlr.D058487"},{"doi-asserted-by":"publisher","key":"e_1_2_10_52_1","DOI":"10.1016\/j.molbiopara.2018.06.002"},{"doi-asserted-by":"publisher","key":"e_1_2_10_53_1","DOI":"10.1016\/b978\u20100\u201012\u2010397925\u20104.00005\u20105"},{"doi-asserted-by":"publisher","key":"e_1_2_10_54_1","DOI":"10.1042\/bj20080795"},{"doi-asserted-by":"publisher","key":"e_1_2_10_55_1","DOI":"10.1038\/cdd.2016.11"}],"container-title":["Analytical Science Advances"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/ansa.202200016","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/ansa.202200016","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/ansa.202200016","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T16:23:36Z","timestamp":1759940616000},"score":1,"resource":{"primary":{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/10.1002\/ansa.202200016"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,8]]},"references-count":54,"journal-issue":{"issue":"7-8","published-print":{"date-parts":[[2022,8]]}},"alternative-id":["10.1002\/ansa.202200016"],"URL":"https:\/\/doi.org\/10.1002\/ansa.202200016","archive":["Portico"],"relation":{},"ISSN":["2628-5452","2628-5452"],"issn-type":[{"type":"print","value":"2628-5452"},{"type":"electronic","value":"2628-5452"}],"subject":[],"published":{"date-parts":[[2022,8]]},"assertion":[{"value":"2022-05-12","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-08-06","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-08-30","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}