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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Mechanism of Fluconazole Resistance in <italic>Candida albicans</italic> Biofilms: Phase-Specific Role of Efflux Pumps and Membrane Sterols </title><author><name sortKey="Mukherjee, Pranab K" sort="Mukherjee, Pranab K" uniqKey="Mukherjee P" first="Pranab K." last="Mukherjee">Pranab K. Mukherjee</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Chandra, Jyotsna" sort="Chandra, Jyotsna" uniqKey="Chandra J" first="Jyotsna" last="Chandra">Jyotsna Chandra</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kuhn, Duncan M" sort="Kuhn, Duncan M" uniqKey="Kuhn D" first="Duncan M." last="Kuhn">Duncan M. Kuhn</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Ghannoum, Mahmoud A" sort="Ghannoum, Mahmoud A" uniqKey="Ghannoum M" first="Mahmoud A." last="Ghannoum">Mahmoud A. Ghannoum</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">12874310</idno><idno type="pmc">165995</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC165995</idno><idno type="RBID">PMC:165995</idno><idno type="doi">10.1128/IAI.71.8.4333-4340.2003</idno><date when="2003">2003</date><idno type="wicri:Area/Pmc/Corpus">001528</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">001528</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Mechanism of Fluconazole Resistance in <italic>Candida albicans</italic> Biofilms: Phase-Specific Role of Efflux Pumps and Membrane Sterols </title><author><name sortKey="Mukherjee, Pranab K" sort="Mukherjee, Pranab K" uniqKey="Mukherjee P" first="Pranab K." last="Mukherjee">Pranab K. Mukherjee</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Chandra, Jyotsna" sort="Chandra, Jyotsna" uniqKey="Chandra J" first="Jyotsna" last="Chandra">Jyotsna Chandra</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kuhn, Duncan M" sort="Kuhn, Duncan M" uniqKey="Kuhn D" first="Duncan M." last="Kuhn">Duncan M. Kuhn</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Ghannoum, Mahmoud A" sort="Ghannoum, Mahmoud A" uniqKey="Ghannoum M" first="Mahmoud A." last="Ghannoum">Mahmoud A. Ghannoum</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">Infection and Immunity</title><idno type="ISSN">0019-9567</idno><idno type="eISSN">1098-5522</idno><imprint><date when="2003">2003</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><italic>Candida albicans</italic> biofilms are formed through three distinct developmental phases and are associated with high fluconazole (FLU) resistance. In the present study, we used a set of isogenic <italic>Candida</italic> strains lacking one or more of the drug efflux pumps Cdr1p, Cdr2p, and Mdr1p to determine their role in FLU resistance of biofilms. Additionally, variation in sterol profile as a possible mechanism of drug resistance was investigated. Our results indicate that parent and mutant strains formed similar biofilms. However, biofilms formed by double and triple mutants were more susceptible to FLU at 6 h (MIC = 64 and 16 μg/ml, respectively) than the wild-type strain (MIC > 256 μg/ml). At later time points (12 and 48 h), all the strains became resistant to this azole (MIC ≥ 256 μg/ml), indicating lack of involvement of efflux pumps in resistance at late stages of biofilm formation. Northern blot analyses revealed that <italic>Candida</italic> biofilms expressed <italic>CDR</italic> and <italic>MDR1</italic> genes in all the developmental phases, while planktonic cells expressed these genes only at the 12- and 48-h time points. Functionality of efflux pumps was assayed by rhodamine (Rh123) efflux assays, which revealed significant differences in Rh123 retention between biofilm and planktonic cells at the early phase (<italic>P</italic> = 0.0006) but not at later stages (12 and 48 h). Sterol analyses showed that ergosterol levels were significantly decreased (<italic>P</italic> < 0.001) at intermediate and mature phases, compared to those in early-phase biofilms. These studies suggest that multicomponent, phase-specific mechanisms are operative in antifungal resistance of fungal biofilms.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Infect Immun</journal-id><journal-id journal-id-type="publisher-id">iai</journal-id><journal-title>Infection and Immunity</journal-title><issn pub-type="ppub">0019-9567</issn><issn pub-type="epub">1098-5522</issn><publisher><publisher-name>American Society for Microbiology</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">12874310</article-id><article-id pub-id-type="pmc">165995</article-id><article-id pub-id-type="publisher-id">0173</article-id><article-id pub-id-type="doi">10.1128/IAI.71.8.4333-4340.2003</article-id><article-categories><subj-group subj-group-type="heading"><subject>Fungal and Parasitic Infections</subject></subj-group></article-categories><title-group><article-title>Mechanism of Fluconazole Resistance in <italic>Candida albicans</italic> Biofilms: Phase-Specific Role of Efflux Pumps and Membrane Sterols </article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Mukherjee</surname><given-names>Pranab K.</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Chandra</surname><given-names>Jyotsna</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Kuhn</surname><given-names>Duncan M.</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="aff" rid="aff1">2</xref></contrib><contrib contrib-type="author"><name><surname>Ghannoum</surname><given-names>Mahmoud A.</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="aff1">Center for Medical Mycology, Department of Dermatology, University Hospitals of Cleveland and Case Western Reserve University,<label>1</label> Division of Infectious Diseases, Department of Medicine, University Hospitals of Cleveland, Cleveland, Ohio 44106<label>2</label></aff><author-notes><fn id="cor1"><label>*</label><p>Corresponding author. Mailing address: Center for Medical Mycology, Department of Dermatology, University Hospitals of Cleveland and Case Western Reserve University, 11100 Euclid Ave., Cleveland, OH 44106-5028. Phone: (216) 844-8580. Fax: (216) 844-1076. E-mail: <email>mag3@cwru.edu</email>.</p></fn></author-notes><pub-date pub-type="ppub"><month>8</month><year>2003</year></pub-date><volume>71</volume><issue>8</issue><fpage>4333</fpage><lpage>4340</lpage><history><date date-type="received"><day>12</day><month>2</month><year>2003</year></date><date date-type="rev-recd"><day>12</day><month>3</month><year>2003</year></date><date date-type="accepted"><day>3</day><month>5</month><year>2003</year></date></history><copyright-statement>Copyright © 2003, American Society for Microbiology</copyright-statement><copyright-year>2003</copyright-year><abstract><p><italic>Candida albicans</italic> biofilms are formed through three distinct developmental phases and are associated with high fluconazole (FLU) resistance. In the present study, we used a set of isogenic <italic>Candida</italic> strains lacking one or more of the drug efflux pumps Cdr1p, Cdr2p, and Mdr1p to determine their role in FLU resistance of biofilms. Additionally, variation in sterol profile as a possible mechanism of drug resistance was investigated. Our results indicate that parent and mutant strains formed similar biofilms. However, biofilms formed by double and triple mutants were more susceptible to FLU at 6 h (MIC = 64 and 16 μg/ml, respectively) than the wild-type strain (MIC > 256 μg/ml). At later time points (12 and 48 h), all the strains became resistant to this azole (MIC ≥ 256 μg/ml), indicating lack of involvement of efflux pumps in resistance at late stages of biofilm formation. Northern blot analyses revealed that <italic>Candida</italic> biofilms expressed <italic>CDR</italic> and <italic>MDR1</italic> genes in all the developmental phases, while planktonic cells expressed these genes only at the 12- and 48-h time points. Functionality of efflux pumps was assayed by rhodamine (Rh123) efflux assays, which revealed significant differences in Rh123 retention between biofilm and planktonic cells at the early phase (<italic>P</italic> = 0.0006) but not at later stages (12 and 48 h). Sterol analyses showed that ergosterol levels were significantly decreased (<italic>P</italic> < 0.001) at intermediate and mature phases, compared to those in early-phase biofilms. These studies suggest that multicomponent, phase-specific mechanisms are operative in antifungal resistance of fungal biofilms.</p></abstract></article-meta><notes><fn-group><fn><p><italic>Editor:</italic> T. R. Kozel</p></fn></fn-group></notes></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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