{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,14]],"date-time":"2026-01-14T14:29:58Z","timestamp":1768400998404,"version":"3.49.0"},"reference-count":32,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2014,8,1]],"date-time":"2014-08-01T00:00:00Z","timestamp":1406851200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2014,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p><jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Candida krusei<\/jats:named-content>is an important agent of opportunistic infections that often displays resistance to several antifungals. We describe here the<jats:italic>in vivo<\/jats:italic>acquisition of resistance to voriconazole (VRC) by<jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">C. krusei<\/jats:named-content>isolates recovered from a leukemia patient during a long period of VRC therapy. In order to mimic the<jats:italic>in vivo<\/jats:italic>development of VRC resistance, a susceptible<jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">C. krusei<\/jats:named-content>isolate was exposed daily to 1 \u03bcg\/ml of VRC<jats:italic>in vitro<\/jats:italic>. Interestingly, after 5 days of exposure to VRC, a MIC of 4 \u03bcg\/ml was achieved; this value remained constant after 25 additional days of treatment with VRC and also after 30 consecutive days of incubation in VRC-free medium. Our objective was to determine the associated molecular resistance mechanisms, such as expression of efflux pump genes and<jats:italic>ERG11<\/jats:italic>gene mutations, among the resistant strains. Synergistic effects between the efflux blocker tacrolimus (FK506) and VRC were found in all of the resistant strains. Moreover,<jats:italic>ABC1<\/jats:italic>gene expression increased over time in both the<jats:italic>in vivo<\/jats:italic>- and<jats:italic>in vitro<\/jats:italic>-induced resistant strains, in contrast to the<jats:italic>ABC2<\/jats:italic>and<jats:italic>ERG11<\/jats:italic>genes, whose expression was invariably lower and constant.<jats:italic>ERG11<\/jats:italic>gene sequencing showed two different types of mutations, i.e., heterozygosity at T1389T\/C, corresponding to synonymous mutations, in<jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">C. krusei<\/jats:named-content>strains and a missense mutation at position T418C, resulting in a change from Tyr to His, among resistant<jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">C. krusei<\/jats:named-content>clinical isolates. This study highlights the relevance of ATP-dependent efflux pump (namely, Abc1p) activity in VRC resistance and describes new mutations in the<jats:italic>ERG11<\/jats:italic>gene among resistant<jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">C. krusei<\/jats:named-content>clinical isolates.<\/jats:p>","DOI":"10.1128\/aac.02603-14","type":"journal-article","created":{"date-parts":[[2014,5,28]],"date-time":"2014-05-28T07:08:08Z","timestamp":1401260888000},"page":"4604-4611","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":35,"title":["<i>In Vivo<\/i>and<i>In Vitro<\/i>Acquisition of Resistance to Voriconazole by Candida krusei"],"prefix":"10.1128","volume":"58","author":[{"given":"Elisabete","family":"Ricardo","sequence":"first","affiliation":[{"name":"Department of Microbiology, Faculty of Medicine, University of Porto, Porto, Portugal"},{"name":"Cardiovascular R&amp;D Unit, Faculty of Medicine, University of Porto, Porto, Portugal"}]},{"given":"Isabel M.","family":"Miranda","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Medicine, University of Porto, Porto, Portugal"},{"name":"Cardiovascular R&amp;D Unit, Faculty of Medicine, University of Porto, Porto, Portugal"},{"name":"CINTESIS\u2014Center for Health Technology and Services Research, Faculty of Medicine, University of Porto, Porto, Portugal"}]},{"given":"Isabel","family":"Faria-Ramos","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Medicine, University of Porto, Porto, Portugal"}]},{"given":"Raquel M.","family":"Silva","sequence":"additional","affiliation":[{"name":"IPATIMUP\u2014Institute of Molecular Pathology and Immunology, University of Porto, Porto, Portugal"}]},{"given":"Ac\u00e1cio Gon\u00e7alves","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Medicine, University of Porto, Porto, Portugal"},{"name":"CINTESIS\u2014Center for Health Technology and Services Research, Faculty of Medicine, University of Porto, Porto, Portugal"}]},{"given":"Cid\u00e1lia","family":"Pina-Vaz","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Medicine, University of Porto, Porto, Portugal"},{"name":"CINTESIS\u2014Center for Health Technology and Services Research, Faculty of Medicine, University of Porto, Porto, Portugal"},{"name":"Department of Microbiology, Hospital S\u00e3o Jo\u00e3o, Porto, Portugal"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1001\/archinte.160.17.2659"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1086\/432060"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01915-07"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkh532"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.41.1.78-83.2003"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10096-004-1174-9"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1086\/507891"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00148-06"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00067-07"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1080\/mmy.39.1.109.116"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.42.10.2645"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.40.11.2443"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01095-08"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.diagmicrobio.2009.11.006"},{"key":"e_1_3_2_16_2","volume-title":"Reference method for broth dilution antifungal susceptibility testing of yeasts: approved standard M27-A3","author":"Clinical and Laboratory Standards Institute","year":"2008","unstructured":"Clinical and Laboratory Standards Institute. 2008. 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