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We have selected an amphotericin B-resistant clinical isolate which demonstrated 8-fold-higher 50% lethal doses (LD\n                    <jats:sub>50<\/jats:sub>\n                    ) than an amphotericin B-sensitive strain to explore the mechanism of amphotericin B resistance. Fluorimetric analysis demonstrated lower anisotropy in the motion of the diphenylhexatriene fluorescent probe in the resistant strain, which indicated a higher fluidity of the membrane for the resistant strain than for the sensitive strain. The expression patterns of the two transcripts of\n                    <jats:italic>S<\/jats:italic>\n                    -adenosyl-\n                    <jats:sc>l<\/jats:sc>\n                    -methionine:C-24-\u0394-sterol methyltransferase and the absence of ergosterol, replaced by cholesta-5,7,24-trien-3\u03b2-ol in the membrane of the resistant parasite, indicate a decreased amphotericin B affinity, which is evidenced by decreased amphotericin B uptake. The expression level of MDR1 is found to be higher in the resistant strain, suggesting a higher rate of efflux of amphotericin B. The resistant parasite also possesses an upregulated tryparedoxin cascade and a more-reduced intracellular thiol level, which helps in better scavenging of reactive oxygen species produced by amphotericin B. The resistance to amphotericin B was partially reverted by the thiol metabolic pathway and ABC transporter inhibitors. Thus, it can be concluded that altered membrane composition, ATP-binding cassette transporters, and an upregulated thiol metabolic pathway have a role in conferring amphotericin B resistance in clinical isolates of\n                    <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Leishmania donovani<\/jats:named-content>\n                    .\n                  <\/jats:p>","DOI":"10.1128\/aac.00030-11","type":"journal-article","created":{"date-parts":[[2011,11,29]],"date-time":"2011-11-29T21:31:59Z","timestamp":1322602319000},"page":"1031-1041","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":298,"title":["Mechanism of Amphotericin B Resistance in Clinical Isolates of Leishmania donovani"],"prefix":"10.1128","volume":"56","author":[{"given":"Bidyut","family":"Purkait","sequence":"first","affiliation":[{"name":"Department of Molecular Biology, Agamkuan, Patna, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashish","family":"Kumar","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Agamkuan, Patna, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nilay","family":"Nandi","sequence":"additional","affiliation":[{"name":"Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna, Bihar, India; UT Southwestern Medical Center, Dallas, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abul Hasan","family":"Sardar","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Agamkuan, Patna, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sushmita","family":"Das","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Agamkuan, Patna, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sudeep","family":"Kumar","sequence":"additional","affiliation":[{"name":"Institute of Global Health, Internal Medicine, University of New Mexico, Albuquerque, New Mexico, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krishna","family":"Pandey","sequence":"additional","affiliation":[{"name":"Department of Clinical Medicine, Agamkuan, Patna, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vidyananda","family":"Ravidas","sequence":"additional","affiliation":[{"name":"Department of Clinical Medicine, Agamkuan, Patna, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manish","family":"Kumar","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Agamkuan, Patna, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tripti","family":"De","sequence":"additional","affiliation":[{"name":"Indian Institute of Chemical Biology, Kolkata, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dharmendra","family":"Singh","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Agamkuan, Patna, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pradeep","family":"Das","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Agamkuan, Patna, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"crossref","first-page":"1201","DOI":"10.1016\/S0006-2952(98)00129-4","article-title":"Involvement of thiol metabolism in resistance to glucantime in Leishmania tropica","volume":"56","author":"Arana FE","year":"1998","unstructured":"AranaFE. 1998. 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