fluconazole Search Results


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  • 99
    Millipore fluconazole
    OMVs protect yeast against synergistic fungicidal activity of polymyxin B in combination with <t>fluconazole:</t> ( a ) Synergistic activities determined by checkerboard assay; ( b ) time-kill assay; ( c ) 24 h growth-curve kinetics for C. albicans incubated with drug alone and drug in combinations with or without OMVs. The kinetics were measured using Varioskan™ LUX reader with measurements at 1 h intervals. The data for experiments (a,b) show means ± SD from two independent experiments carried out in triplicate; the data for experiment (c) show mean values from two independent repetitions carried out in duplicate; for better readability, SDs were not included.
    Fluconazole, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1280 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    LKT Laboratories fluconazole
    Structures of the triazoles voriconazole, <t>fluconazole</t> and itraconazole and the BR biosynthesis inhibitor Brz2001.
    Fluconazole, supplied by LKT Laboratories, used in various techniques. Bioz Stars score: 93/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Selleck Chemicals fluconazole
    ADA2 control of antifungal drug tolerance and cell wall integrity is mediated by ERG6 . The erg6 mutants were susceptible to echinocandins, <t>fluconazole,</t> cell wall-perturbing agents, and the ER stress chemical tunicamycin but not amphotericin B. Cells were grown overnight in YPD at 37°C and washed twice with dH 2 O, 5-fold serially diluted, and spotted onto YPD medium containing MCF, CSF, ANF, FLC, PSC, VRC, SDS, CFW, CR, DTT, or TM at the indicated concentration. Strains were spotted onto SC medium with or without 50 ng/ml AmB to test tolerance to amphotericin B. All plates were incubated at 37°C for 30 h.
    Fluconazole, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Beckman Coulter zonal rotor
    ADA2 control of antifungal drug tolerance and cell wall integrity is mediated by ERG6 . The erg6 mutants were susceptible to echinocandins, <t>fluconazole,</t> cell wall-perturbing agents, and the ER stress chemical tunicamycin but not amphotericin B. Cells were grown overnight in YPD at 37°C and washed twice with dH 2 O, 5-fold serially diluted, and spotted onto YPD medium containing MCF, CSF, ANF, FLC, PSC, VRC, SDS, CFW, CR, DTT, or TM at the indicated concentration. Strains were spotted onto SC medium with or without 50 ng/ml AmB to test tolerance to amphotericin B. All plates were incubated at 37°C for 30 h.
    Zonal Rotor, supplied by Beckman Coulter, used in various techniques. Bioz Stars score: 92/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Cayman Chemical fluconazole
    In vitro evaluation of DB766-azole combinations against intracellular L. donovani . Posaconazole (POS; gray bars), ketoconazole (KTC; horizontal striped bars), or <t>fluconazole</t> (FLC; diagonal striped bars) was added to a serial dilution of DB766, with the azole at a fixed concentration, in at least three independent experiments. The concentrations of azole drugs employed were below those required for 50% inhibition alone (parasite burden was reduced by 39% ± 3% at 6.3 μM posaconazole alone [ n = 3], 23% ± 8% at 25 μM ketoconazole alone [ n = 3], and 14% ± 5% at 50 μM fluconazole alone [ n = 4]). Error bars and measurements represent the standard errors of the means. Two-sided Student's t test was used to compare the groups to the DB766-alone group. **, P
    Fluconazole, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Enzo Biochem fluconazole
    Dectin-1 is required for CD4 + T-cell survival. ( a ) The frequency of apoptotic CD4 + T-cells was measured by Annexin-V staining in mLN cell suspensions at the indicated time points post infection (WT n =9, KO n =9 at each time point). ( b ) CD8 + T-cell apoptosis and necrosis was measured by Annexin-V/7-AAD staining in mLN cell suspensions at 72 h post infection ( n =6). ( c ) TUNEL staining performed on paraformaldehyde-fixed mLNs from infected WT and KO mice 72 h post infection; photo taken at 5 × magnification, insert at 40 × magnification. Blue=DAPI, green=apoptotic cells. ( d ) The total number of recoverable cells from the mLN (WT n =9, KO n =9 at each time point). ( e ) H E staining of mLNs at 72 h post infection compared with naive controls (scale bars are 500 μm). ( f ) The total number of visible ( > 1 mm 3 ) Peyer's patches (infected n =9, naive n =8) in WT and Dectin-1 −/− mice at 72 h post infection. ( g ) The total number of recoverable cells from the mLN of infected WT ( n =10) and KO ( n =9) mice at 3 days post infection while maintained on cholestyramine diets, as described in Figure 1 . ( h ) Schematic of the colitis model where co-housed WT and Dectin-1 KO animals were maintained on water supplemented with penicillin, streptomycin, and <t>fluconazole</t> for 4 days, and then switched to water containing 3.5% DSS and C. tropicalis for 5 days. The number of ( i ) Peyer's patches and ( j ) number of DCs (CD11c + MHCII + ) isolated from mLNs from WT ( n =20) and Dectin-1 KO ( n =16) mice at 5 days post DSS exposure was compared with Candida only WT animals (control, n =6). Bar charts show pooled data (2–5 experiments); overlaid dot plots show one representative experiment. * P
    Fluconazole, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 92/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    HiMedia Laboratories fluconazole
    Antifungal Susceptibility Testing (AFST) of vaginal Candida isolates against <t>fluconazole.</t>
    Fluconazole, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 93/100, based on 219 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Hospira fluconazole
    Nanoscale mixed-species biofilm of S. aureus and C. albicans on chip. (A) Fluorescence micrographs of a section of the spot containing mixed-species nanobiofilms stained with FUN-1 and concanavalin A. Staining with FUN-1 demonstrated all viable fungal and bacterial populations (in orange-yellow), and concanavalin A stained only fungal cell walls (in blue). (B to D) Profile of susceptibility of mixed-species biofilms of S. aureus and C. albicans to antibiotics (B), antifungals (C), and combination treatment (D). The data represent dose-response profiles of mixed-species biofilms with respect to ciprofloxacin, vancomycin, tobramycin, and methicillin (B) and to amphotericin B and <t>fluconazole</t> (C) at 50, 5, 0.5, 0.05, and 0.005 μg/ml. (D) Profile of susceptibility to combinations of 25 μg/ml of vancomycin (VANC) with 25, 2.5, 0.25, 0.025, and 0.0025 μg/ml of amphotericin B (AMB) and fluconazole (FLU).
    Fluconazole, supplied by Hospira, used in various techniques. Bioz Stars score: 90/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Liofilchem fluconazole
    ABDD antifungal susceptibility testing of dermatophytes showing resistance to <t>Fluconazole</t> and Griseofulvin.
    Fluconazole, supplied by Liofilchem, used in various techniques. Bioz Stars score: 92/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Medi Corp fluconazole
    In vitro release of <t>fluconazole</t> from multilamellar liposomes composed of phosphatidylcholine:cholesterol:α-tocopherol acetate (1:0.8:0.1) in phosphate buffered saline.
    Fluconazole, supplied by Medi Corp, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Pfizer Inc fluconazole
    SEM images of 48-h C. albicans HK1Sa. (A) Control C. albicans HK1Sa; (B) C. albicans HK1Sa biofilm exposed to 600-μg/ml amphotericin B for 4 h; (C) C. albicans HK1Sa biofilm exposed to 600-μg/ml <t>fluconazole</t> for 4 h. Note the wrinkled,
    Fluconazole, supplied by Pfizer Inc, used in various techniques. Bioz Stars score: 93/100, based on 1252 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    TargetMol fluconazole
    SEM images of 48-h C. albicans HK1Sa. (A) Control C. albicans HK1Sa; (B) C. albicans HK1Sa biofilm exposed to 600-μg/ml amphotericin B for 4 h; (C) C. albicans HK1Sa biofilm exposed to 600-μg/ml <t>fluconazole</t> for 4 h. Note the wrinkled,
    Fluconazole, supplied by TargetMol, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    ICN Biomedicals fluconazole
    CYP51 inhibitors and catalytic reaction. A , structural formulas of the marketed drugs, 1,2,4-triazoles voriconazole ( Vor ), <t>fluconazole</t> ( Fluc ), and posaconazole ( Poz ), and an experimental CYP51 inhibitor, the 1,3-imidazole VNI. The heme-coordinating nitrogen
    Fluconazole, supplied by ICN Biomedicals, used in various techniques. Bioz Stars score: 91/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    InterChem fluconazole
    Multicomponent therapeutics that prevent proliferation of <t>fluconazole-resistant</t> C. albicans . All 435 possible two-component combinations of 30 compounds were tested in duplicate in 36-point dose matrices to identify synergistic combinations. C. albicans cells were treated with compounds in 384-well plates and the extent of inhibition of Alamar blue fluorescence was determined. ( A ) Overview of synergistic combinations in C. albicans Alamar blue proliferation assay. Yellow columns and rows indicate compounds that are commonly used as antifungal agents. Purple squares indicate combinations for which neither compound is used as an antifungal agent on its own and red indicates combinations for which one, but not both, of the compounds is used as an antifungal agent on its own. ( B ) A sample combination dose matrix, showing the combined effect of pentamidine and phenazopyridine at six concentrations (including a zero concentration point) each. The experimentally measured inhibition of Alamar blue signal is shown for each pair of combinations. The color of the squares also indicates the level of Alamar blue inhibition. ( C ) The calculated excess inhibition over the predicted Bliss additivism model. The predicted Bliss additive effect (see text) was subtracted from the experimentally observed inhibition at each pair of concentrations. ( D ) The calculated excess inhibition over the HSA.
    Fluconazole, supplied by InterChem, used in various techniques. Bioz Stars score: 92/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Mantena Laboratories Ltd fluconazole
    IgG isotypes production against rPb27 by infected and immunized mice. Antibody response against rPb27 in mice infected and immunized with this recombinant protein associated or not with <t>fluconazole</t> chemotherapy was determined by ELISA assay after 40 (A) and 90 (B) days of treatment. Control, mice without any intervention. rPb27, group infected and posteriorly immunized with rPb27. rPb27/T, same as rPb27, but treated with fluconazole. Bars represent the means and standard deviations of optical density (O.D.) at 1∶400 serum dilution in each experimental group (n = 3). * significant (p
    Fluconazole, supplied by Mantena Laboratories Ltd, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Sequoia Research fluconazole
    Beauvericin inhibits Hsp90 function ( a ) Beauvericin (BEA) reduces <t>fluconazole-induced</t> calcineurin activation. S. cerevisiae harboring 4xCDRE- lacZ reporter construct ± fluconazole (FLC, 64 μg/ml), ± geldanamycin (GdA, 5.6 μg/ml), FK506 (1.0 μg/ml), or beauvericin (20 μg/ml). Data are mean ± s.d. from technical triplicates and representative of biological replicates. * P
    Fluconazole, supplied by Sequoia Research, used in various techniques. Bioz Stars score: 93/100, based on 58 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher fluconazole
    Diploid and Tetraploid C.albicans are sensitive to antifungals. A) Diploid colony forming units (CFUs) following 24-hr exposure to 1μg/mL or 10μg/mL <t>fluconazole</t> (‘FLU’, light and dark purple bars) and 0.25μg/mL or 2.5μg/mL caspofungin (‘CAS’, light and dark green bars) treatments. Bars represent the mean of 3 independent experiments (black symbols), and the error bars indicate +/− SEM. The dashed line and shaded box represent the mean and +/− SEM of the no-drug treatment. Asterisks indicate drug treatments that differ significantly from the no-drug treatment (* P
    Fluconazole, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Tokyo Chemical Industry fluconazole
    SDS-PAGE and phosphatase analysis of the mobility shift of hyperactive Tac1. (A) Immunoblot of Tac1 SDS-PAGE mobility in lysates from a 6His3Flag-tagged wild-type Tac1 strain (yLM485) treated with 10 μg/ml FNZ, 10 μg/ml EST, or 40 μg/ml <t>fluconazole</t> (FLC) for the indicated periods of time. Samples were resolved on 6% SDS-PAGE gel and the blot was probed with an anti-Flag antibody. Coomassie blue staining (CBS) was used as the loading control. (B) Immunoblot analysis of SDS-PAGE mobility in lysates from strains expressing an endogenous level of individual 6His3Flag-tagged Tac1 variants (wild-type [yLM485], N972D [yLM534], Δ962–969 [yLM535], ΔM677 [yLM533], R693K [yLM532], and A736V [yLM531]) in the absence and presence of 10 μg/ml fluphenazine. Samples were resolved on 6% SDS-PAGE gel and the blot was probed with an anti-Flag antibody. CBS was used as the loading control. (C) Immunoblot analysis demonstrating the effect of λ protein phosphatase (λ-pp) treatment on the SDS-PAGE mobility shift of hyperactive Tac1 purified from strains that overexpressed 6His3Flag (HF)-tagged wild-type Tac1 (yLM530; grown in the presence and absence of fluphenazine), Tac1 R693K (yLM540), or Tac1 N977D (yLM536). Samples were resolved by 6% SDS-PAGE and the blot was probed by an anti-Flag antibody. (D) Immunoblot analysis of the effect of λ protein phosphatase treatment (in the presence and absence of 50 mM NaF and 20 mM Na 3 VO 4 [F − /VO 4 3− ] phosphatase inhibitors) on the SDS-PAGE mobility shift of Tac1 purified from 6His3Flag-tagged wild-type Tac1 cells (yLM530) grown in the absence and presence of fluphenazine. Samples were resolved by 6% SDS-PAGE and the blot was probed by an anti-Flag antibody.
    Fluconazole, supplied by Tokyo Chemical Industry, used in various techniques. Bioz Stars score: 92/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    OMVs protect yeast against synergistic fungicidal activity of polymyxin B in combination with fluconazole: ( a ) Synergistic activities determined by checkerboard assay; ( b ) time-kill assay; ( c ) 24 h growth-curve kinetics for C. albicans incubated with drug alone and drug in combinations with or without OMVs. The kinetics were measured using Varioskan™ LUX reader with measurements at 1 h intervals. The data for experiments (a,b) show means ± SD from two independent experiments carried out in triplicate; the data for experiment (c) show mean values from two independent repetitions carried out in duplicate; for better readability, SDs were not included.

    Journal: International Journal of Molecular Sciences

    Article Title: Interspecies Outer Membrane Vesicles (OMVs) Modulate the Sensitivity of Pathogenic Bacteria and Pathogenic Yeasts to Cationic Peptides and Serum Complement

    doi: 10.3390/ijms20225577

    Figure Lengend Snippet: OMVs protect yeast against synergistic fungicidal activity of polymyxin B in combination with fluconazole: ( a ) Synergistic activities determined by checkerboard assay; ( b ) time-kill assay; ( c ) 24 h growth-curve kinetics for C. albicans incubated with drug alone and drug in combinations with or without OMVs. The kinetics were measured using Varioskan™ LUX reader with measurements at 1 h intervals. The data for experiments (a,b) show means ± SD from two independent experiments carried out in triplicate; the data for experiment (c) show mean values from two independent repetitions carried out in duplicate; for better readability, SDs were not included.

    Article Snippet: Reagents BHI (Brain Heart Infusion, OXOID, Basingstoke, UK) ); TSB (Tryptone Soya Broth, OXOID, Hampshire, England); TSA (Tryptone Soya Agar, OXOID, Hampshire, England), Bradford reagent (Protein Assay Dye Reagent Concentrate, Bio-Rad, München, Germany); β-nicotinoamide adenine dinucleotide hydrate (NAD, Sigma-Aldrich, Steinheim, Germany); fluconazole (FLC, Sigma-Aldrich, Poznan, Poland), hemin (Sigma-Aldrich, St. Louis, MO, USA); polymyxin B sulfate salt (PB, Sigma-Aldrich, Denmark); fluorescein isothiocyanate (FITC, ThermoScientific, Rockford, IL, USA); Hank’s Buffer with Ca2+ , Mg2+ (HBSS, PAN Biotech, UK); RPMI 1640 (Lonza, Walkersville, MD, USA); o -nitrophenyl-β-D-galactopyranoside (ONPG, Sigma, Steinheim Germany), heat inactivated (56 °C, 1 h) FBS (Fetal bovine serum, Gibco Life Technologies, Grand Island, NY, USA).).

    Techniques: Activity Assay, Time-Kill Assay, Incubation

    Cell viability curve and IC 50 of the P. salutare essential oil (A, C and E) and the oil in combined with fluconazole (B, D and F) against different Candida spp. strains, at different collection periods. Concentration of fluconazole: 2,048 µg/mL. OEFPs, Essential oil of the leaves of Psidium salutare , 1, 2 and 3 collections; CA, C. albicans ; CT, C. tropicalis ; CK, C. krusei ; INCQS, National Institute of Quality Control in Health. (A) Cell viability curve and IC 50 of Psidium salutare essential oil against Candida albicans . (B) Cell viability curve and IC 50 of Psidium salutare essential oil combined with fluconazole against Candida albicans . (C) Cell viability curve and IC 50 of Psidium salutare essential oil against Candida tropicalis . (D) Cell viability curve and IC 50 of Psidium salutare essential oil combined with fluconazole against Candida tropicalis . (E) Cell viability curve and IC 50 of Psidium salutare essential oil against Candida krusei . (F) Cell viability curve and IC 50 of Psidium salutare essential oil combined with fluconazole against Candida krusei .

    Journal: PeerJ

    Article Title: Effect of seasonality on chemical profile and antifungal activity of essential oil isolated from leaves Psidium salutare (Kunth) O. Berg

    doi: 10.7717/peerj.5476

    Figure Lengend Snippet: Cell viability curve and IC 50 of the P. salutare essential oil (A, C and E) and the oil in combined with fluconazole (B, D and F) against different Candida spp. strains, at different collection periods. Concentration of fluconazole: 2,048 µg/mL. OEFPs, Essential oil of the leaves of Psidium salutare , 1, 2 and 3 collections; CA, C. albicans ; CT, C. tropicalis ; CK, C. krusei ; INCQS, National Institute of Quality Control in Health. (A) Cell viability curve and IC 50 of Psidium salutare essential oil against Candida albicans . (B) Cell viability curve and IC 50 of Psidium salutare essential oil combined with fluconazole against Candida albicans . (C) Cell viability curve and IC 50 of Psidium salutare essential oil against Candida tropicalis . (D) Cell viability curve and IC 50 of Psidium salutare essential oil combined with fluconazole against Candida tropicalis . (E) Cell viability curve and IC 50 of Psidium salutare essential oil against Candida krusei . (F) Cell viability curve and IC 50 of Psidium salutare essential oil combined with fluconazole against Candida krusei .

    Article Snippet: Both the essential oil and antifungal fluconazole (F8929 ≥ 98% (HPLC), powder; Sigma Aldrich, St. Louis, MO, USA) was previously diluted in dimethylsulfoxide (DMSO; Dynamic, Indaiatuba, Brazil) and its final concentration was adjusted with addition of distilled water to obtain the desired concentration for (16,384 µg / ml).

    Techniques: Concentration Assay

    Effect of the Psidium salutare essential oil on Candida albicans yeast micromorphological aspects. Culture performed in depleted potato dextrose agar medium, with 40×objective visualization. (A) Growth control, (B) fluconazole antifungal effect at 2,048 µg/mL, (C) P. salutare essential oil effect at 8,192 μg/mL, (D) P. salutare essential oil effect at 2,048 µg/mL and (E) (C) P. salutare essential oil effect at 512 µg/mL; CA, C. albicans ; INCQS, National Institute of Quality Control in Health.

    Journal: PeerJ

    Article Title: Effect of seasonality on chemical profile and antifungal activity of essential oil isolated from leaves Psidium salutare (Kunth) O. Berg

    doi: 10.7717/peerj.5476

    Figure Lengend Snippet: Effect of the Psidium salutare essential oil on Candida albicans yeast micromorphological aspects. Culture performed in depleted potato dextrose agar medium, with 40×objective visualization. (A) Growth control, (B) fluconazole antifungal effect at 2,048 µg/mL, (C) P. salutare essential oil effect at 8,192 μg/mL, (D) P. salutare essential oil effect at 2,048 µg/mL and (E) (C) P. salutare essential oil effect at 512 µg/mL; CA, C. albicans ; INCQS, National Institute of Quality Control in Health.

    Article Snippet: Both the essential oil and antifungal fluconazole (F8929 ≥ 98% (HPLC), powder; Sigma Aldrich, St. Louis, MO, USA) was previously diluted in dimethylsulfoxide (DMSO; Dynamic, Indaiatuba, Brazil) and its final concentration was adjusted with addition of distilled water to obtain the desired concentration for (16,384 µg / ml).

    Techniques:

    FLC protect the epithelial cells from the damage of C. albicans in vitro. a Cell damage reflected by LDH release after 24 h post infection compared to wild type (WT) stain without FLC (1 × 10 5 cells per mL). b Reactive oxygen species (ROS) released by TR146 epithelial cells after infected with WT strain with or without FLC (fold change to 0 min). c IL-1α production at 24 h post infection, 1 × 10 4 cells per mL. d Obviously decreased adhesion to TR146 epithelial cells by WT strain with FLC after 60 min. FLC fluconazole, LDH lactate dehydrogenase, IL interleukin

    Journal: International Journal of Oral Science

    Article Title: ERG3 and ERG11 genes are critical for the pathogenesis of Candida albicans during the oral mucosal infection

    doi: 10.1038/s41368-018-0013-2

    Figure Lengend Snippet: FLC protect the epithelial cells from the damage of C. albicans in vitro. a Cell damage reflected by LDH release after 24 h post infection compared to wild type (WT) stain without FLC (1 × 10 5 cells per mL). b Reactive oxygen species (ROS) released by TR146 epithelial cells after infected with WT strain with or without FLC (fold change to 0 min). c IL-1α production at 24 h post infection, 1 × 10 4 cells per mL. d Obviously decreased adhesion to TR146 epithelial cells by WT strain with FLC after 60 min. FLC fluconazole, LDH lactate dehydrogenase, IL interleukin

    Article Snippet: Chemicals Fluconazole (98.5%) was commercially obtained from Sigma-Aldrich (China) and dissolved with dimethylsulfoxide (DMSO, Merck-China).

    Techniques: In Vitro, Infection, Staining

    FLC can cure the epithelial infection caused by C. albicans at low dosage in vivo. a Images of infected mice tongues with oral candidal leukoplakia after 2-day oropharyngeal infection with wild type (WT) strain drinking water with or without different doses of FLC. Leukoplakia on tongue are indicated in vivo by black arrow while showed by white arrow on incided tissue. b Fungal burdens obtained from the tongues of mice after 2-day oropharyngeal infection with C. albicans WT strain with or without FLC. c PAS- and HE-stained tongues from mice 2 days post infection by C. albicans . Including whole-mount and high-magnification views infected by WT with or without different doses of FLC. Invading hyphae are indicated by black arrowhead and inflammatory cells are showed by blue arrowhead. d Average percentage of the mice entire tongue epithelium area infected by WT strain. e Susceptibility of C. albicans to macrophagocyte with or without different doses of FLC. fluconazole, PAS Periodic Acid-Schiff, HE hematein eosin

    Journal: International Journal of Oral Science

    Article Title: ERG3 and ERG11 genes are critical for the pathogenesis of Candida albicans during the oral mucosal infection

    doi: 10.1038/s41368-018-0013-2

    Figure Lengend Snippet: FLC can cure the epithelial infection caused by C. albicans at low dosage in vivo. a Images of infected mice tongues with oral candidal leukoplakia after 2-day oropharyngeal infection with wild type (WT) strain drinking water with or without different doses of FLC. Leukoplakia on tongue are indicated in vivo by black arrow while showed by white arrow on incided tissue. b Fungal burdens obtained from the tongues of mice after 2-day oropharyngeal infection with C. albicans WT strain with or without FLC. c PAS- and HE-stained tongues from mice 2 days post infection by C. albicans . Including whole-mount and high-magnification views infected by WT with or without different doses of FLC. Invading hyphae are indicated by black arrowhead and inflammatory cells are showed by blue arrowhead. d Average percentage of the mice entire tongue epithelium area infected by WT strain. e Susceptibility of C. albicans to macrophagocyte with or without different doses of FLC. fluconazole, PAS Periodic Acid-Schiff, HE hematein eosin

    Article Snippet: Chemicals Fluconazole (98.5%) was commercially obtained from Sigma-Aldrich (China) and dissolved with dimethylsulfoxide (DMSO, Merck-China).

    Techniques: Infection, In Vivo, Mouse Assay, Staining

    Sequential therapy time-kill curve for C . albicans (a) and C . guillermondii (b) in the presence of amphotericin B or fluconazole (concentration = 0.25 x MIC) after 12 h pre exposure to C32 (concentration = 0.25 x MIC). Color coding is provided in the Figure. Standard deviations are omitted for clarity. Normally, these were between 0.1 to 0.5 Log CFU ml -1 units.

    Journal: PLoS ONE

    Article Title: Antibiotic saving effect of combination therapy through synergistic interactions between well-characterized chito-oligosaccharides and commercial antifungals against medically relevant yeasts

    doi: 10.1371/journal.pone.0227098

    Figure Lengend Snippet: Sequential therapy time-kill curve for C . albicans (a) and C . guillermondii (b) in the presence of amphotericin B or fluconazole (concentration = 0.25 x MIC) after 12 h pre exposure to C32 (concentration = 0.25 x MIC). Color coding is provided in the Figure. Standard deviations are omitted for clarity. Normally, these were between 0.1 to 0.5 Log CFU ml -1 units.

    Article Snippet: Antifungals Commercial antifungals (CA), fluconazole (Flu), amphotericin B (Amp), voriconazole (Vor), flucytosine (Fcs), and miconazole (Mcz), were purchased from Sigma (St. Louis, MO).

    Techniques: Concentration Assay

    Fluconazole sensitivity of strains expressing hyperactive MRR1 and CAP1 alleles in wild-type or ada2 Δ backgrounds. Serial 10-fold dilutions of the strains were spotted onto SD agar plates with or without 5 μg/ml fluconazole and incubated

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: SAGA/ADA Complex Subunit Ada2 Is Required for Cap1- but Not Mrr1-Mediated Upregulation of the Candida albicans Multidrug Efflux Pump MDR1

    doi: 10.1128/AAC.03065-14

    Figure Lengend Snippet: Fluconazole sensitivity of strains expressing hyperactive MRR1 and CAP1 alleles in wild-type or ada2 Δ backgrounds. Serial 10-fold dilutions of the strains were spotted onto SD agar plates with or without 5 μg/ml fluconazole and incubated

    Article Snippet: To determine the fluconazole susceptibilities of the strains, a 2-fold dilution series of fluconazole (Sigma GmbH, Deisenhofen, Germany) was prepared in the assay medium, starting from an initial concentration of 128 μg/ml.

    Techniques: Expressing, Incubation

    Sensitivity of the wild type, heterozygous and homozygous ada2 Δ mutants, and complemented strains to fluconazole, H 2 O 2 , and nourseothricin. Serial 10-fold dilutions of the strains were spotted onto agar plates containing the indicated compounds.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: SAGA/ADA Complex Subunit Ada2 Is Required for Cap1- but Not Mrr1-Mediated Upregulation of the Candida albicans Multidrug Efflux Pump MDR1

    doi: 10.1128/AAC.03065-14

    Figure Lengend Snippet: Sensitivity of the wild type, heterozygous and homozygous ada2 Δ mutants, and complemented strains to fluconazole, H 2 O 2 , and nourseothricin. Serial 10-fold dilutions of the strains were spotted onto agar plates containing the indicated compounds.

    Article Snippet: To determine the fluconazole susceptibilities of the strains, a 2-fold dilution series of fluconazole (Sigma GmbH, Deisenhofen, Germany) was prepared in the assay medium, starting from an initial concentration of 128 μg/ml.

    Techniques:

    Structures of the triazoles voriconazole, fluconazole and itraconazole and the BR biosynthesis inhibitor Brz2001.

    Journal: PLoS ONE

    Article Title: Genetic Variation in Plant CYP51s Confers Resistance against Voriconazole, a Novel Inhibitor of Brassinosteroid-Dependent Sterol Biosynthesis

    doi: 10.1371/journal.pone.0053650

    Figure Lengend Snippet: Structures of the triazoles voriconazole, fluconazole and itraconazole and the BR biosynthesis inhibitor Brz2001.

    Article Snippet: Chemicals Bifonazole, fluconazole, itraconazole, thiabendazole and uniconazole were purchased from LKT Laboratories (St. Paul, MN, USA).

    Techniques:

    Voriconazole, itraconazole and fluconazole induce phenotypes indicative of BR deficiency in arabidopsis and cress. ( A ) Arabidopsis plants grown on plates containing 5 µM of the indicated compounds under long-day conditions for 7 days. ( B ) Hypocotyl length of cress seedlings grown for 3 days on plates containing different concentrations of the indicated compounds. Data are the mean ± SD of 30 seedlings measured.

    Journal: PLoS ONE

    Article Title: Genetic Variation in Plant CYP51s Confers Resistance against Voriconazole, a Novel Inhibitor of Brassinosteroid-Dependent Sterol Biosynthesis

    doi: 10.1371/journal.pone.0053650

    Figure Lengend Snippet: Voriconazole, itraconazole and fluconazole induce phenotypes indicative of BR deficiency in arabidopsis and cress. ( A ) Arabidopsis plants grown on plates containing 5 µM of the indicated compounds under long-day conditions for 7 days. ( B ) Hypocotyl length of cress seedlings grown for 3 days on plates containing different concentrations of the indicated compounds. Data are the mean ± SD of 30 seedlings measured.

    Article Snippet: Chemicals Bifonazole, fluconazole, itraconazole, thiabendazole and uniconazole were purchased from LKT Laboratories (St. Paul, MN, USA).

    Techniques:

    ADA2 control of antifungal drug tolerance and cell wall integrity is mediated by ERG6 . The erg6 mutants were susceptible to echinocandins, fluconazole, cell wall-perturbing agents, and the ER stress chemical tunicamycin but not amphotericin B. Cells were grown overnight in YPD at 37°C and washed twice with dH 2 O, 5-fold serially diluted, and spotted onto YPD medium containing MCF, CSF, ANF, FLC, PSC, VRC, SDS, CFW, CR, DTT, or TM at the indicated concentration. Strains were spotted onto SC medium with or without 50 ng/ml AmB to test tolerance to amphotericin B. All plates were incubated at 37°C for 30 h.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Deletion of ADA2 Increases Antifungal Drug Susceptibility and Virulence in Candida glabrata

    doi: 10.1128/AAC.01924-17

    Figure Lengend Snippet: ADA2 control of antifungal drug tolerance and cell wall integrity is mediated by ERG6 . The erg6 mutants were susceptible to echinocandins, fluconazole, cell wall-perturbing agents, and the ER stress chemical tunicamycin but not amphotericin B. Cells were grown overnight in YPD at 37°C and washed twice with dH 2 O, 5-fold serially diluted, and spotted onto YPD medium containing MCF, CSF, ANF, FLC, PSC, VRC, SDS, CFW, CR, DTT, or TM at the indicated concentration. Strains were spotted onto SC medium with or without 50 ng/ml AmB to test tolerance to amphotericin B. All plates were incubated at 37°C for 30 h.

    Article Snippet: Sodium dodecyl sulfate (SDS; Bioman, New Taipei City, Taiwan), calcofluor white (CFW; Fluorescent Brighter 28; Sigma), Congo red (CR; Genzyme, Cambridge, MA, USA), fluconazole (FLC; Selleckchem, Houston, TX, USA), posaconazole (PSC; Merck, Rahway, NJ, USA), voriconazole (VRC; Sigma), micafungin (MCF; Astellas Pharma Inc., Deerfield, IL, USA), caspofungin (CSF; Merck), anidulafungin (ANF; Pfizer Inc., Groton, CT, USA), dithiothreitol (DTT; BioShop, Burlington, ON, Canada), and tunicamycin (TM, Sigma) were added to the media at the concentrations indicated above and below.

    Techniques: Concentration Assay, Incubation

    In vitro evaluation of DB766-azole combinations against intracellular L. donovani . Posaconazole (POS; gray bars), ketoconazole (KTC; horizontal striped bars), or fluconazole (FLC; diagonal striped bars) was added to a serial dilution of DB766, with the azole at a fixed concentration, in at least three independent experiments. The concentrations of azole drugs employed were below those required for 50% inhibition alone (parasite burden was reduced by 39% ± 3% at 6.3 μM posaconazole alone [ n = 3], 23% ± 8% at 25 μM ketoconazole alone [ n = 3], and 14% ± 5% at 50 μM fluconazole alone [ n = 4]). Error bars and measurements represent the standard errors of the means. Two-sided Student's t test was used to compare the groups to the DB766-alone group. **, P

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Antileishmanial Efficacy and Pharmacokinetics of DB766-Azole Combinations

    doi: 10.1128/AAC.01129-17

    Figure Lengend Snippet: In vitro evaluation of DB766-azole combinations against intracellular L. donovani . Posaconazole (POS; gray bars), ketoconazole (KTC; horizontal striped bars), or fluconazole (FLC; diagonal striped bars) was added to a serial dilution of DB766, with the azole at a fixed concentration, in at least three independent experiments. The concentrations of azole drugs employed were below those required for 50% inhibition alone (parasite burden was reduced by 39% ± 3% at 6.3 μM posaconazole alone [ n = 3], 23% ± 8% at 25 μM ketoconazole alone [ n = 3], and 14% ± 5% at 50 μM fluconazole alone [ n = 4]). Error bars and measurements represent the standard errors of the means. Two-sided Student's t test was used to compare the groups to the DB766-alone group. **, P

    Article Snippet: Azoles were obtained from the following sources: ketoconazole was from TCI (Portland, OR), posaconazole was from Carbosynth (San Diego, CA), and fluconazole was from Cayman Chemical (Ann Arbor, MI).

    Techniques: In Vitro, Serial Dilution, Concentration Assay, Inhibition

    Structures of DB766, posaconazole, ketoconazole, and fluconazole.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Antileishmanial Efficacy and Pharmacokinetics of DB766-Azole Combinations

    doi: 10.1128/AAC.01129-17

    Figure Lengend Snippet: Structures of DB766, posaconazole, ketoconazole, and fluconazole.

    Article Snippet: Azoles were obtained from the following sources: ketoconazole was from TCI (Portland, OR), posaconazole was from Carbosynth (San Diego, CA), and fluconazole was from Cayman Chemical (Ann Arbor, MI).

    Techniques:

    Dectin-1 is required for CD4 + T-cell survival. ( a ) The frequency of apoptotic CD4 + T-cells was measured by Annexin-V staining in mLN cell suspensions at the indicated time points post infection (WT n =9, KO n =9 at each time point). ( b ) CD8 + T-cell apoptosis and necrosis was measured by Annexin-V/7-AAD staining in mLN cell suspensions at 72 h post infection ( n =6). ( c ) TUNEL staining performed on paraformaldehyde-fixed mLNs from infected WT and KO mice 72 h post infection; photo taken at 5 × magnification, insert at 40 × magnification. Blue=DAPI, green=apoptotic cells. ( d ) The total number of recoverable cells from the mLN (WT n =9, KO n =9 at each time point). ( e ) H E staining of mLNs at 72 h post infection compared with naive controls (scale bars are 500 μm). ( f ) The total number of visible ( > 1 mm 3 ) Peyer's patches (infected n =9, naive n =8) in WT and Dectin-1 −/− mice at 72 h post infection. ( g ) The total number of recoverable cells from the mLN of infected WT ( n =10) and KO ( n =9) mice at 3 days post infection while maintained on cholestyramine diets, as described in Figure 1 . ( h ) Schematic of the colitis model where co-housed WT and Dectin-1 KO animals were maintained on water supplemented with penicillin, streptomycin, and fluconazole for 4 days, and then switched to water containing 3.5% DSS and C. tropicalis for 5 days. The number of ( i ) Peyer's patches and ( j ) number of DCs (CD11c + MHCII + ) isolated from mLNs from WT ( n =20) and Dectin-1 KO ( n =16) mice at 5 days post DSS exposure was compared with Candida only WT animals (control, n =6). Bar charts show pooled data (2–5 experiments); overlaid dot plots show one representative experiment. * P

    Journal: Mucosal Immunology

    Article Title: CD4+ T-cell survival in the GI tract requires dectin-1 during fungal infection

    doi: 10.1038/mi.2015.79

    Figure Lengend Snippet: Dectin-1 is required for CD4 + T-cell survival. ( a ) The frequency of apoptotic CD4 + T-cells was measured by Annexin-V staining in mLN cell suspensions at the indicated time points post infection (WT n =9, KO n =9 at each time point). ( b ) CD8 + T-cell apoptosis and necrosis was measured by Annexin-V/7-AAD staining in mLN cell suspensions at 72 h post infection ( n =6). ( c ) TUNEL staining performed on paraformaldehyde-fixed mLNs from infected WT and KO mice 72 h post infection; photo taken at 5 × magnification, insert at 40 × magnification. Blue=DAPI, green=apoptotic cells. ( d ) The total number of recoverable cells from the mLN (WT n =9, KO n =9 at each time point). ( e ) H E staining of mLNs at 72 h post infection compared with naive controls (scale bars are 500 μm). ( f ) The total number of visible ( > 1 mm 3 ) Peyer's patches (infected n =9, naive n =8) in WT and Dectin-1 −/− mice at 72 h post infection. ( g ) The total number of recoverable cells from the mLN of infected WT ( n =10) and KO ( n =9) mice at 3 days post infection while maintained on cholestyramine diets, as described in Figure 1 . ( h ) Schematic of the colitis model where co-housed WT and Dectin-1 KO animals were maintained on water supplemented with penicillin, streptomycin, and fluconazole for 4 days, and then switched to water containing 3.5% DSS and C. tropicalis for 5 days. The number of ( i ) Peyer's patches and ( j ) number of DCs (CD11c + MHCII + ) isolated from mLNs from WT ( n =20) and Dectin-1 KO ( n =16) mice at 5 days post DSS exposure was compared with Candida only WT animals (control, n =6). Bar charts show pooled data (2–5 experiments); overlaid dot plots show one representative experiment. * P

    Article Snippet: Colitis model. Co-housed WT and Dectin-1−/− animals were maintained on sterile water supplemented with 2 mg ml−1 streptomycin, 2,000 U ml−1 penicillin (both Invitrogen) and 0.25 mg ml−1 fluconazole (Enzo, Exeter, UK) for 4 days prior to start of experiments.

    Techniques: Staining, Infection, TUNEL Assay, Mouse Assay, Isolation

    Antifungal Susceptibility Testing (AFST) of vaginal Candida isolates against fluconazole.

    Journal: Journal of Clinical and Diagnostic Research : JCDR

    Article Title: Fungal Profile of Vulvovaginal Candidiasis in a Tertiary Care Hospital

    doi: 10.7860/JCDR/2017/23578.9475

    Figure Lengend Snippet: Antifungal Susceptibility Testing (AFST) of vaginal Candida isolates against fluconazole.

    Article Snippet: Antifungal susceptibility tests were performed by the disc diffusion method using fluconazole (25 μg) and voriconazole (1 μg) (Hi-Media, Mumbai, India), on Muller Hinton Agar (MHA) (Hi-Media, Mumbai, India) supplemented with 2% Glucose and Methylene Blue dye 0.5 μg/ml (GMB) as per the CLSI-M44-A2 guidelines [ ].

    Techniques:

    Nanoscale mixed-species biofilm of S. aureus and C. albicans on chip. (A) Fluorescence micrographs of a section of the spot containing mixed-species nanobiofilms stained with FUN-1 and concanavalin A. Staining with FUN-1 demonstrated all viable fungal and bacterial populations (in orange-yellow), and concanavalin A stained only fungal cell walls (in blue). (B to D) Profile of susceptibility of mixed-species biofilms of S. aureus and C. albicans to antibiotics (B), antifungals (C), and combination treatment (D). The data represent dose-response profiles of mixed-species biofilms with respect to ciprofloxacin, vancomycin, tobramycin, and methicillin (B) and to amphotericin B and fluconazole (C) at 50, 5, 0.5, 0.05, and 0.005 μg/ml. (D) Profile of susceptibility to combinations of 25 μg/ml of vancomycin (VANC) with 25, 2.5, 0.25, 0.025, and 0.0025 μg/ml of amphotericin B (AMB) and fluconazole (FLU).

    Journal: mSphere

    Article Title: nBioChip, a Lab-on-a-Chip Platform of Mono- and Polymicrobial Biofilms for High-Throughput Downstream Applications

    doi: 10.1128/mSphere.00247-17

    Figure Lengend Snippet: Nanoscale mixed-species biofilm of S. aureus and C. albicans on chip. (A) Fluorescence micrographs of a section of the spot containing mixed-species nanobiofilms stained with FUN-1 and concanavalin A. Staining with FUN-1 demonstrated all viable fungal and bacterial populations (in orange-yellow), and concanavalin A stained only fungal cell walls (in blue). (B to D) Profile of susceptibility of mixed-species biofilms of S. aureus and C. albicans to antibiotics (B), antifungals (C), and combination treatment (D). The data represent dose-response profiles of mixed-species biofilms with respect to ciprofloxacin, vancomycin, tobramycin, and methicillin (B) and to amphotericin B and fluconazole (C) at 50, 5, 0.5, 0.05, and 0.005 μg/ml. (D) Profile of susceptibility to combinations of 25 μg/ml of vancomycin (VANC) with 25, 2.5, 0.25, 0.025, and 0.0025 μg/ml of amphotericin B (AMB) and fluconazole (FLU).

    Article Snippet: Another iteration of printing was carried out to deposit 50 nl of amphotericin B (Sigma, MO) and fluconazole (Hospira, IL) at dilutions of 50, 5, 0.5, 0.05, and 0.005 μg/ml.

    Techniques: Chromatin Immunoprecipitation, Fluorescence, Staining

    ABDD antifungal susceptibility testing of dermatophytes showing resistance to Fluconazole and Griseofulvin.

    Journal: Electronic Physician

    Article Title: Isolation, Identification, and In Vitro Antifungal Susceptibility Testing of Dermatophytes from Clinical Samples at Sohag University Hospital in Egypt

    doi: 10.19082/2557

    Figure Lengend Snippet: ABDD antifungal susceptibility testing of dermatophytes showing resistance to Fluconazole and Griseofulvin.

    Article Snippet: Antifungal susceptibility testing ABDD antifungal susceptibility testing was performed using four antifungal agents: Clotrimazole (50 μg), Miconazole (10 μg), Fluconazole (25μg), and Griseofulvin (10 μg) (Liofilchem diagnostic, Italy).

    Techniques:

    In vitro release of fluconazole from multilamellar liposomes composed of phosphatidylcholine:cholesterol:α-tocopherol acetate (1:0.8:0.1) in phosphate buffered saline.

    Journal: Saudi Pharmaceutical Journal : SPJ

    Article Title: Effect of formulation design and freeze-drying on properties of fluconazole multilamellar liposomes

    doi: 10.1016/j.jsps.2010.07.003

    Figure Lengend Snippet: In vitro release of fluconazole from multilamellar liposomes composed of phosphatidylcholine:cholesterol:α-tocopherol acetate (1:0.8:0.1) in phosphate buffered saline.

    Article Snippet: Fluconazole was obtained from Medicorp (city, India) as a gift sample. l -α-Phosphatidylcholine (PC), type X-E from dried egg yolk, cholesterol (Chol), stearylamine (SA), dicetyl phosphate (DP), β-glucose, d -trehalose and β-lactose were purchased from Sigma Chemical Co. (St. Louis, USA). α-Tocopherol acetate was purchased from La Roche (France).

    Techniques: In Vitro

    3.5. Freeze-drying of fluconazole liposomes

    Journal: Saudi Pharmaceutical Journal : SPJ

    Article Title: Effect of formulation design and freeze-drying on properties of fluconazole multilamellar liposomes

    doi: 10.1016/j.jsps.2010.07.003

    Figure Lengend Snippet: 3.5. Freeze-drying of fluconazole liposomes

    Article Snippet: Fluconazole was obtained from Medicorp (city, India) as a gift sample. l -α-Phosphatidylcholine (PC), type X-E from dried egg yolk, cholesterol (Chol), stearylamine (SA), dicetyl phosphate (DP), β-glucose, d -trehalose and β-lactose were purchased from Sigma Chemical Co. (St. Louis, USA). α-Tocopherol acetate was purchased from La Roche (France).

    Techniques:

    In vitro release of fluconazole from multilamellar liposomes composed of Phosphatidylcholine:cholesterol:α-tocopherol acetate (1:0.6:0.1) in phosphate buffered saline.

    Journal: Saudi Pharmaceutical Journal : SPJ

    Article Title: Effect of formulation design and freeze-drying on properties of fluconazole multilamellar liposomes

    doi: 10.1016/j.jsps.2010.07.003

    Figure Lengend Snippet: In vitro release of fluconazole from multilamellar liposomes composed of Phosphatidylcholine:cholesterol:α-tocopherol acetate (1:0.6:0.1) in phosphate buffered saline.

    Article Snippet: Fluconazole was obtained from Medicorp (city, India) as a gift sample. l -α-Phosphatidylcholine (PC), type X-E from dried egg yolk, cholesterol (Chol), stearylamine (SA), dicetyl phosphate (DP), β-glucose, d -trehalose and β-lactose were purchased from Sigma Chemical Co. (St. Louis, USA). α-Tocopherol acetate was purchased from La Roche (France).

    Techniques: In Vitro

    SEM images of 48-h C. albicans HK1Sa. (A) Control C. albicans HK1Sa; (B) C. albicans HK1Sa biofilm exposed to 600-μg/ml amphotericin B for 4 h; (C) C. albicans HK1Sa biofilm exposed to 600-μg/ml fluconazole for 4 h. Note the wrinkled,

    Journal:

    Article Title: In Vitro Method To Study Antifungal Perfusion in Candida Biofilms

    doi: 10.1128/JCM.43.2.818-825.2005

    Figure Lengend Snippet: SEM images of 48-h C. albicans HK1Sa. (A) Control C. albicans HK1Sa; (B) C. albicans HK1Sa biofilm exposed to 600-μg/ml amphotericin B for 4 h; (C) C. albicans HK1Sa biofilm exposed to 600-μg/ml fluconazole for 4 h. Note the wrinkled,

    Article Snippet: Three antifungals commonly used to treat oropharyngeal and systemic candidiasis were selected for the study, viz., amphotericin B (Sigma), fluconazole (Pfizer), and flucytosine (Sigma).

    Techniques:

    Standard curves for the three antifungal agents, showing the relationship between drug concentration and the radius of growth inhibition of a lawn of C. parapsilosis on RPMI agar. 5FC, flucytosine; FL, fluconazole; AmB, amphotericin B.

    Journal:

    Article Title: In Vitro Method To Study Antifungal Perfusion in Candida Biofilms

    doi: 10.1128/JCM.43.2.818-825.2005

    Figure Lengend Snippet: Standard curves for the three antifungal agents, showing the relationship between drug concentration and the radius of growth inhibition of a lawn of C. parapsilosis on RPMI agar. 5FC, flucytosine; FL, fluconazole; AmB, amphotericin B.

    Article Snippet: Three antifungals commonly used to treat oropharyngeal and systemic candidiasis were selected for the study, viz., amphotericin B (Sigma), fluconazole (Pfizer), and flucytosine (Sigma).

    Techniques: Concentration Assay, Inhibition

    Penetration of various concentrations (i.e., 150, 300, and 600 μg/ml) of the three antifungals, amphotericin B (a), fluconazole (b), and flucytosine (c), through 48-h-old C. albicans , C. parapsilosis , and C. krusei biofilms shown in terms of the

    Journal:

    Article Title: In Vitro Method To Study Antifungal Perfusion in Candida Biofilms

    doi: 10.1128/JCM.43.2.818-825.2005

    Figure Lengend Snippet: Penetration of various concentrations (i.e., 150, 300, and 600 μg/ml) of the three antifungals, amphotericin B (a), fluconazole (b), and flucytosine (c), through 48-h-old C. albicans , C. parapsilosis , and C. krusei biofilms shown in terms of the

    Article Snippet: Three antifungals commonly used to treat oropharyngeal and systemic candidiasis were selected for the study, viz., amphotericin B (Sigma), fluconazole (Pfizer), and flucytosine (Sigma).

    Techniques:

    SEM images of 48-h C. krusei ATCC 6258. (A) Control C. krusei ATCC 6258; (B) C. krusei ATCC 6258 biofilm exposed to 600-μg/ml amphotericin B for 4 h; (C) C. krusei ATCC 6258 biofilm exposed to 600-μg/ml fluconazole for 4 h.

    Journal:

    Article Title: In Vitro Method To Study Antifungal Perfusion in Candida Biofilms

    doi: 10.1128/JCM.43.2.818-825.2005

    Figure Lengend Snippet: SEM images of 48-h C. krusei ATCC 6258. (A) Control C. krusei ATCC 6258; (B) C. krusei ATCC 6258 biofilm exposed to 600-μg/ml amphotericin B for 4 h; (C) C. krusei ATCC 6258 biofilm exposed to 600-μg/ml fluconazole for 4 h.

    Article Snippet: Three antifungals commonly used to treat oropharyngeal and systemic candidiasis were selected for the study, viz., amphotericin B (Sigma), fluconazole (Pfizer), and flucytosine (Sigma).

    Techniques:

    SEM images of 48-h C. parapsilosis ATCC 22019. (A) Control; (B) C. parapsilosis ATCC 22019 biofilm exposed to 600-μg/ml amphotericin B for 4 h; (C) C. parapsilosis ATCC 22019 biofilm exposed to 600-μg/ml fluconazole for 4 h. Note the ruptured

    Journal:

    Article Title: In Vitro Method To Study Antifungal Perfusion in Candida Biofilms

    doi: 10.1128/JCM.43.2.818-825.2005

    Figure Lengend Snippet: SEM images of 48-h C. parapsilosis ATCC 22019. (A) Control; (B) C. parapsilosis ATCC 22019 biofilm exposed to 600-μg/ml amphotericin B for 4 h; (C) C. parapsilosis ATCC 22019 biofilm exposed to 600-μg/ml fluconazole for 4 h. Note the ruptured

    Article Snippet: Three antifungals commonly used to treat oropharyngeal and systemic candidiasis were selected for the study, viz., amphotericin B (Sigma), fluconazole (Pfizer), and flucytosine (Sigma).

    Techniques:

    CYP51 inhibitors and catalytic reaction. A , structural formulas of the marketed drugs, 1,2,4-triazoles voriconazole ( Vor ), fluconazole ( Fluc ), and posaconazole ( Poz ), and an experimental CYP51 inhibitor, the 1,3-imidazole VNI. The heme-coordinating nitrogen

    Journal: The Journal of Biological Chemistry

    Article Title: Structure-Functional Characterization of Cytochrome P450 Sterol 14α-Demethylase (CYP51B) from Aspergillus fumigatus and Molecular Basis for the Development of Antifungal Drugs *

    doi: 10.1074/jbc.M115.677310

    Figure Lengend Snippet: CYP51 inhibitors and catalytic reaction. A , structural formulas of the marketed drugs, 1,2,4-triazoles voriconazole ( Vor ), fluconazole ( Fluc ), and posaconazole ( Poz ), and an experimental CYP51 inhibitor, the 1,3-imidazole VNI. The heme-coordinating nitrogen

    Article Snippet: Voriconazole, ketoconazole, itraconazole, and posaconazole were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX); fluconazole, clotrimazole, and miconazole were from ICN Biomedicals.

    Techniques:

    Multicomponent therapeutics that prevent proliferation of fluconazole-resistant C. albicans . All 435 possible two-component combinations of 30 compounds were tested in duplicate in 36-point dose matrices to identify synergistic combinations. C. albicans cells were treated with compounds in 384-well plates and the extent of inhibition of Alamar blue fluorescence was determined. ( A ) Overview of synergistic combinations in C. albicans Alamar blue proliferation assay. Yellow columns and rows indicate compounds that are commonly used as antifungal agents. Purple squares indicate combinations for which neither compound is used as an antifungal agent on its own and red indicates combinations for which one, but not both, of the compounds is used as an antifungal agent on its own. ( B ) A sample combination dose matrix, showing the combined effect of pentamidine and phenazopyridine at six concentrations (including a zero concentration point) each. The experimentally measured inhibition of Alamar blue signal is shown for each pair of combinations. The color of the squares also indicates the level of Alamar blue inhibition. ( C ) The calculated excess inhibition over the predicted Bliss additivism model. The predicted Bliss additive effect (see text) was subtracted from the experimentally observed inhibition at each pair of concentrations. ( D ) The calculated excess inhibition over the HSA.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Systematic discovery of multicomponent therapeutics

    doi: 10.1073/pnas.1337088100

    Figure Lengend Snippet: Multicomponent therapeutics that prevent proliferation of fluconazole-resistant C. albicans . All 435 possible two-component combinations of 30 compounds were tested in duplicate in 36-point dose matrices to identify synergistic combinations. C. albicans cells were treated with compounds in 384-well plates and the extent of inhibition of Alamar blue fluorescence was determined. ( A ) Overview of synergistic combinations in C. albicans Alamar blue proliferation assay. Yellow columns and rows indicate compounds that are commonly used as antifungal agents. Purple squares indicate combinations for which neither compound is used as an antifungal agent on its own and red indicates combinations for which one, but not both, of the compounds is used as an antifungal agent on its own. ( B ) A sample combination dose matrix, showing the combined effect of pentamidine and phenazopyridine at six concentrations (including a zero concentration point) each. The experimentally measured inhibition of Alamar blue signal is shown for each pair of combinations. The color of the squares also indicates the level of Alamar blue inhibition. ( C ) The calculated excess inhibition over the predicted Bliss additivism model. The predicted Bliss additive effect (see text) was subtracted from the experimentally observed inhibition at each pair of concentrations. ( D ) The calculated excess inhibition over the HSA.

    Article Snippet: Fluconazole (Interchem, Paramus, NJ) and phenazopyridine (Sigma) were additionally tested at the indicated concentrations for their effect on the proliferation of fluconazole-resistant C. albicans (strain 17; Seattle Biomedical Research Institute, Seattle).

    Techniques: Inhibition, Fluorescence, Proliferation Assay, Concentration Assay

    The combination of fluconazole and phenazopyridine selectively inhibits proliferation of fluconazole-resistant C. albicans .( A ) The percent inhibition of fluconazole-resistant C. albicans proliferation is shown for the indicated concentrations of fluconazole and phenazopyridine, determined by using an Alamar blue proliferation assay. The average of three measurements is shown. ( B ) The calculated excess inhibition over the Bliss additivism model. The predicted Bliss additive effect (see text) was subtracted from the experimentally observed inhibition at each pair of concentrations. ( C ) The calculated excess inhibition over the HSA. ( D ) Percent inhibition of fluconazole-resistant C. albicans proliferation at concentrations showing optimal synergy [250 μM fluconazole (flu) and/or 20 μM phenazolepyridine (PAP)]. ( E ) Fungicidal activity, determined by using a cfu assay. Fluconazole-resistant C. albicans were treated with 20 μM PAP and/or 250 μM flu, or 4 μM amphotericin B (amp B) as a fungicidal positive control, and in each case an equal number of yeast particles were plated in the absence of any compound. The number of colonies that grew after each treatment regime is indicated. ( F ) The combination of PAP and flu prevents dye efflux. Fluconazoleresistant C. albicans cells were treated with 20 μM PAP and/or 250 μM flu and the effect on efflux of rhodamine G was determined by using fluorescence microscopy. Phase-contrast images of the yeast cells are shown in each case to confirm that a large number of cells were present in each case, but that dye efflux was only effectively inhibited when both PAP and flu were present. For all numerical figures shown, the average of three measurements is represented. Error bars, 1 SD.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Systematic discovery of multicomponent therapeutics

    doi: 10.1073/pnas.1337088100

    Figure Lengend Snippet: The combination of fluconazole and phenazopyridine selectively inhibits proliferation of fluconazole-resistant C. albicans .( A ) The percent inhibition of fluconazole-resistant C. albicans proliferation is shown for the indicated concentrations of fluconazole and phenazopyridine, determined by using an Alamar blue proliferation assay. The average of three measurements is shown. ( B ) The calculated excess inhibition over the Bliss additivism model. The predicted Bliss additive effect (see text) was subtracted from the experimentally observed inhibition at each pair of concentrations. ( C ) The calculated excess inhibition over the HSA. ( D ) Percent inhibition of fluconazole-resistant C. albicans proliferation at concentrations showing optimal synergy [250 μM fluconazole (flu) and/or 20 μM phenazolepyridine (PAP)]. ( E ) Fungicidal activity, determined by using a cfu assay. Fluconazole-resistant C. albicans were treated with 20 μM PAP and/or 250 μM flu, or 4 μM amphotericin B (amp B) as a fungicidal positive control, and in each case an equal number of yeast particles were plated in the absence of any compound. The number of colonies that grew after each treatment regime is indicated. ( F ) The combination of PAP and flu prevents dye efflux. Fluconazoleresistant C. albicans cells were treated with 20 μM PAP and/or 250 μM flu and the effect on efflux of rhodamine G was determined by using fluorescence microscopy. Phase-contrast images of the yeast cells are shown in each case to confirm that a large number of cells were present in each case, but that dye efflux was only effectively inhibited when both PAP and flu were present. For all numerical figures shown, the average of three measurements is represented. Error bars, 1 SD.

    Article Snippet: Fluconazole (Interchem, Paramus, NJ) and phenazopyridine (Sigma) were additionally tested at the indicated concentrations for their effect on the proliferation of fluconazole-resistant C. albicans (strain 17; Seattle Biomedical Research Institute, Seattle).

    Techniques: Inhibition, Proliferation Assay, Activity Assay, Colony-forming Unit Assay, Positive Control, Fluorescence, Microscopy

    IgG isotypes production against rPb27 by infected and immunized mice. Antibody response against rPb27 in mice infected and immunized with this recombinant protein associated or not with fluconazole chemotherapy was determined by ELISA assay after 40 (A) and 90 (B) days of treatment. Control, mice without any intervention. rPb27, group infected and posteriorly immunized with rPb27. rPb27/T, same as rPb27, but treated with fluconazole. Bars represent the means and standard deviations of optical density (O.D.) at 1∶400 serum dilution in each experimental group (n = 3). * significant (p

    Journal: PLoS ONE

    Article Title: Additive Effect of rPb27 Immunization and Chemotherapy in Experimental Paracoccidioidomycosis

    doi: 10.1371/journal.pone.0017885

    Figure Lengend Snippet: IgG isotypes production against rPb27 by infected and immunized mice. Antibody response against rPb27 in mice infected and immunized with this recombinant protein associated or not with fluconazole chemotherapy was determined by ELISA assay after 40 (A) and 90 (B) days of treatment. Control, mice without any intervention. rPb27, group infected and posteriorly immunized with rPb27. rPb27/T, same as rPb27, but treated with fluconazole. Bars represent the means and standard deviations of optical density (O.D.) at 1∶400 serum dilution in each experimental group (n = 3). * significant (p

    Article Snippet: Chemotherapy of infected mice After 30 days of infection animals were treated for 30 days during which groups of mice received doses every 24 h of fluconazole 10 mg/Kg (Mantena laboratories limited).

    Techniques: Infection, Mouse Assay, Recombinant, Enzyme-linked Immunosorbent Assay

    Representative histopathology of lungs, livers and spleens from infected mice, after 40 days of treatment. BALB/c mice were euthanized 40 days after treatment. The lungs, spleens, and livers were excised, fixed in 10% buffered formalin, and embedded in paraffin for sectioning. The sections were stained with hematoxylin–eosin and examined microscopically. Infected, group only infected with P. brasiliensis . Infected/T, same as Infected, but treated with fluconazole. rPb27, group infected and posteriorly immunized with rPb27. rPb27/T, same as rPb27, but treated with fluconazole. In each lung photos, the scale bar represents 427.3 µm, while in each liver and spleen photos, the scale bar represents 56.9 µm.

    Journal: PLoS ONE

    Article Title: Additive Effect of rPb27 Immunization and Chemotherapy in Experimental Paracoccidioidomycosis

    doi: 10.1371/journal.pone.0017885

    Figure Lengend Snippet: Representative histopathology of lungs, livers and spleens from infected mice, after 40 days of treatment. BALB/c mice were euthanized 40 days after treatment. The lungs, spleens, and livers were excised, fixed in 10% buffered formalin, and embedded in paraffin for sectioning. The sections were stained with hematoxylin–eosin and examined microscopically. Infected, group only infected with P. brasiliensis . Infected/T, same as Infected, but treated with fluconazole. rPb27, group infected and posteriorly immunized with rPb27. rPb27/T, same as rPb27, but treated with fluconazole. In each lung photos, the scale bar represents 427.3 µm, while in each liver and spleen photos, the scale bar represents 56.9 µm.

    Article Snippet: Chemotherapy of infected mice After 30 days of infection animals were treated for 30 days during which groups of mice received doses every 24 h of fluconazole 10 mg/Kg (Mantena laboratories limited).

    Techniques: Histopathology, Infection, Mouse Assay, Staining

    Fungal recovery in lung, spleen and liver of infected mice. The CFUs were estimated 40 (A) and 90 days (B) post treatment in organs from mice infected intratracheally with 3×10 5 P. brasiliensis yeast cells and subjected to fluconazole treatment combined or not with rPb27 immunization. Control mice were only infected with P. brasiliensis (Infected), adjuvant mice were inoculated with C. parvum -Al(OH) 3 with fluconazole treatment (Adjuvant/T) or not (Adjuvant), and rPb27 mice were immunized with recombinant protein combined to fluconazole treatment (rPb27/T) or not (rPb27). All groups of mice were infected with the same number of yeast cells. Bars represent the Log 10 (UFC/g) means and standard deviations from organs of 3 to 5 animals in each group. * significant (p

    Journal: PLoS ONE

    Article Title: Additive Effect of rPb27 Immunization and Chemotherapy in Experimental Paracoccidioidomycosis

    doi: 10.1371/journal.pone.0017885

    Figure Lengend Snippet: Fungal recovery in lung, spleen and liver of infected mice. The CFUs were estimated 40 (A) and 90 days (B) post treatment in organs from mice infected intratracheally with 3×10 5 P. brasiliensis yeast cells and subjected to fluconazole treatment combined or not with rPb27 immunization. Control mice were only infected with P. brasiliensis (Infected), adjuvant mice were inoculated with C. parvum -Al(OH) 3 with fluconazole treatment (Adjuvant/T) or not (Adjuvant), and rPb27 mice were immunized with recombinant protein combined to fluconazole treatment (rPb27/T) or not (rPb27). All groups of mice were infected with the same number of yeast cells. Bars represent the Log 10 (UFC/g) means and standard deviations from organs of 3 to 5 animals in each group. * significant (p

    Article Snippet: Chemotherapy of infected mice After 30 days of infection animals were treated for 30 days during which groups of mice received doses every 24 h of fluconazole 10 mg/Kg (Mantena laboratories limited).

    Techniques: Infection, Mouse Assay, Recombinant

    Representative granulome histopathology of lungs from infected mice after 40 and 90 days of infection. BALB/c mice were euthanized 40 and 90 days after treatment. The lungs, spleens, and livers were excised, fixed in 10% buffered formalin, and embedded in paraffin for sectioning. The sections were stained with hematoxylin–eosin and examined microscopically. Infected, group only infected with P. brasiliensis . Infected/T, same as Infected, but treated with fluconazole. rPb27/T, group infected and posteriorly immunized with rPb27 and treated with fluconazole. In each photo, the scale bar represents 25.9 µm.

    Journal: PLoS ONE

    Article Title: Additive Effect of rPb27 Immunization and Chemotherapy in Experimental Paracoccidioidomycosis

    doi: 10.1371/journal.pone.0017885

    Figure Lengend Snippet: Representative granulome histopathology of lungs from infected mice after 40 and 90 days of infection. BALB/c mice were euthanized 40 and 90 days after treatment. The lungs, spleens, and livers were excised, fixed in 10% buffered formalin, and embedded in paraffin for sectioning. The sections were stained with hematoxylin–eosin and examined microscopically. Infected, group only infected with P. brasiliensis . Infected/T, same as Infected, but treated with fluconazole. rPb27/T, group infected and posteriorly immunized with rPb27 and treated with fluconazole. In each photo, the scale bar represents 25.9 µm.

    Article Snippet: Chemotherapy of infected mice After 30 days of infection animals were treated for 30 days during which groups of mice received doses every 24 h of fluconazole 10 mg/Kg (Mantena laboratories limited).

    Techniques: Histopathology, Infection, Mouse Assay, Staining

    Representative histopathology of lungs, livers and spleens from infected mice, after 90 days of treatment. BALB/c mice were euthanized 90 days after treatment. The lungs, spleens, and livers were excised, fixed in 10% buffered formalin, and embedded in paraffin for sectioning. The sections were stained with hematoxylin–eosin and examined microscopically. Infected, group only infected with P. brasiliensis . Infected/T, same as Infected, but treated with fluconazole. rPb27, group infected and posteriorly immunized with rPb27. rPb27/T, same as rPb27, but treated with fluconazole. In each lung photos, the scale bar represents 416.6 µm, while in each liver and spleen photos, the scale bar represents 55.6 µm.

    Journal: PLoS ONE

    Article Title: Additive Effect of rPb27 Immunization and Chemotherapy in Experimental Paracoccidioidomycosis

    doi: 10.1371/journal.pone.0017885

    Figure Lengend Snippet: Representative histopathology of lungs, livers and spleens from infected mice, after 90 days of treatment. BALB/c mice were euthanized 90 days after treatment. The lungs, spleens, and livers were excised, fixed in 10% buffered formalin, and embedded in paraffin for sectioning. The sections were stained with hematoxylin–eosin and examined microscopically. Infected, group only infected with P. brasiliensis . Infected/T, same as Infected, but treated with fluconazole. rPb27, group infected and posteriorly immunized with rPb27. rPb27/T, same as rPb27, but treated with fluconazole. In each lung photos, the scale bar represents 416.6 µm, while in each liver and spleen photos, the scale bar represents 55.6 µm.

    Article Snippet: Chemotherapy of infected mice After 30 days of infection animals were treated for 30 days during which groups of mice received doses every 24 h of fluconazole 10 mg/Kg (Mantena laboratories limited).

    Techniques: Histopathology, Infection, Mouse Assay, Staining

    Beauvericin inhibits Hsp90 function ( a ) Beauvericin (BEA) reduces fluconazole-induced calcineurin activation. S. cerevisiae harboring 4xCDRE- lacZ reporter construct ± fluconazole (FLC, 64 μg/ml), ± geldanamycin (GdA, 5.6 μg/ml), FK506 (1.0 μg/ml), or beauvericin (20 μg/ml). Data are mean ± s.d. from technical triplicates and representative of biological replicates. * P

    Journal: Nature chemical biology

    Article Title: Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling

    doi: 10.1038/nchembio.2165

    Figure Lengend Snippet: Beauvericin inhibits Hsp90 function ( a ) Beauvericin (BEA) reduces fluconazole-induced calcineurin activation. S. cerevisiae harboring 4xCDRE- lacZ reporter construct ± fluconazole (FLC, 64 μg/ml), ± geldanamycin (GdA, 5.6 μg/ml), FK506 (1.0 μg/ml), or beauvericin (20 μg/ml). Data are mean ± s.d. from technical triplicates and representative of biological replicates. * P

    Article Snippet: Fluconazole (Sequoia Research Products) was dissolved in sterile ddH2 O, beauvericin in 100% methanol, cycloheximide (AG Scientific Inc.) in sterile ddH2 O, rhodamine-6G (Sigma-Aldrich) in sterile ddH2 O, geldanamycin (Invitrogen) in DMSO, and cyclosporin A (CalBiochem) in DMSO.

    Techniques: Activation Assay, Construct

    Beauvericin inhibits Pdr5, thereby increasing fluconazole intracellular accumulation, and can be effluxed by Pdr5 following substitutions that alter substrate-specificity ( a ) S. cerevisiae lacking Yor1 is sensitive to beauvericin, but acquires resistance via Pdr5 substitutions. 1×10 8 cells plated on YEPD containing beauvericin (100 μg/ml); resistant mutants recovered after 3–4 days at 30°C. Resistance was assessed in YEPD with two-fold serial dilutions of beauvericin, as in Figure 1 . Mutations identified by genome sequencing indicated as amino acid changes; those in Pdr5 shown in red. ( b ) Functional validation of PDR5 mutations. Amino acid 538 is altered in three of five mutants; expression of PDR5 G538R in yor1 Δ pdr5 Δ confers beauvericin resistance. Resistance was assessed in SD medium at 30°C for 72 hours. (a–b) Performed in biological triplicates with technical duplicates. ( c ) Beauvericin-resistant mutants exhibit altered substrate specificity, and beauvericin enhances azole accumulation. Deletion of PDR5 increased intracellular accumulation of radiolabelled fluconazole, as with beauvericin-resistant mutants yor1 ΔR1 and R5. Error bars represent standard deviation (s.d.), * P

    Journal: Nature chemical biology

    Article Title: Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling

    doi: 10.1038/nchembio.2165

    Figure Lengend Snippet: Beauvericin inhibits Pdr5, thereby increasing fluconazole intracellular accumulation, and can be effluxed by Pdr5 following substitutions that alter substrate-specificity ( a ) S. cerevisiae lacking Yor1 is sensitive to beauvericin, but acquires resistance via Pdr5 substitutions. 1×10 8 cells plated on YEPD containing beauvericin (100 μg/ml); resistant mutants recovered after 3–4 days at 30°C. Resistance was assessed in YEPD with two-fold serial dilutions of beauvericin, as in Figure 1 . Mutations identified by genome sequencing indicated as amino acid changes; those in Pdr5 shown in red. ( b ) Functional validation of PDR5 mutations. Amino acid 538 is altered in three of five mutants; expression of PDR5 G538R in yor1 Δ pdr5 Δ confers beauvericin resistance. Resistance was assessed in SD medium at 30°C for 72 hours. (a–b) Performed in biological triplicates with technical duplicates. ( c ) Beauvericin-resistant mutants exhibit altered substrate specificity, and beauvericin enhances azole accumulation. Deletion of PDR5 increased intracellular accumulation of radiolabelled fluconazole, as with beauvericin-resistant mutants yor1 ΔR1 and R5. Error bars represent standard deviation (s.d.), * P

    Article Snippet: Fluconazole (Sequoia Research Products) was dissolved in sterile ddH2 O, beauvericin in 100% methanol, cycloheximide (AG Scientific Inc.) in sterile ddH2 O, rhodamine-6G (Sigma-Aldrich) in sterile ddH2 O, geldanamycin (Invitrogen) in DMSO, and cyclosporin A (CalBiochem) in DMSO.

    Techniques: Sequencing, Functional Assay, Expressing, Standard Deviation

    The combination of beauvericin and fluconazole provides a powerful therapeutic strategy ( a ) Beauvericin enhances fluconazole efficacy in a mouse model of C. albicans disseminated infection. Mice were infected with C. albicans (SC5314) and treated with vehicle, beauvericin, fluconazole, or the combination. Even with a high C. albicans inoculum, the combination of beauvericin and fluconazole significantly enhanced survival relative to either treatment alone. Each group consisted of 10 female BALB/c mice. Log-rank (Mantel-Cox) test: vehicle vs. beauvericin: P =0.3173, vehicle vs. fluconazole: P

    Journal: Nature chemical biology

    Article Title: Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling

    doi: 10.1038/nchembio.2165

    Figure Lengend Snippet: The combination of beauvericin and fluconazole provides a powerful therapeutic strategy ( a ) Beauvericin enhances fluconazole efficacy in a mouse model of C. albicans disseminated infection. Mice were infected with C. albicans (SC5314) and treated with vehicle, beauvericin, fluconazole, or the combination. Even with a high C. albicans inoculum, the combination of beauvericin and fluconazole significantly enhanced survival relative to either treatment alone. Each group consisted of 10 female BALB/c mice. Log-rank (Mantel-Cox) test: vehicle vs. beauvericin: P =0.3173, vehicle vs. fluconazole: P

    Article Snippet: Fluconazole (Sequoia Research Products) was dissolved in sterile ddH2 O, beauvericin in 100% methanol, cycloheximide (AG Scientific Inc.) in sterile ddH2 O, rhodamine-6G (Sigma-Aldrich) in sterile ddH2 O, geldanamycin (Invitrogen) in DMSO, and cyclosporin A (CalBiochem) in DMSO.

    Techniques: Infection, Mouse Assay

    Beauvericin enhances fluconazole efficacy against diverse fungi and blocks the emergence of resistance ( a ) Beauvericin (BEA) reduces fluconazole resistance of reference strains of S. cerevisiae (BY4741), C. albicans (SN95), and C. neoformans (H99a), and A. fumigatus clinical isolate on rich medium (YEPD). Fluconazole (FLC) strips generate a gradient from 0.016 to 256 μg/ml, with the highest concentration at the top. Where indicated, plates contain 20 μg/ml of BEA. Experiment performed in biological triplicates. ( b ) BEA and FLC are synergistic and cidal against C. albicans (SN95). SN95 was subjected to two-fold serial dilutions of BEA and FLC in YEPD at 30°C for 48 hours. Optical densities were standardized to drug-free controls (see color bar) and FICI was calculated based on concentrations causing 60–70% growth inhibition. The drug combination is cidal, based on transferring cells to drug-free YEPD medium for 24 hours. ( c ) BEA reduced FLC resistance of C. albicans clinical isolates (CaCi) similar to geldanamycin (GdA) and cyclosporin A (CsA). CaCi are sequentially ordered with isolates recovered early at the top. MIC assays were performed in YEPD (−) with GdA (5 μM), CsA (20 μM), or BEA (25 μM), with a two-fold serial dilution of FLC. (b–c) Experiments performed in biological triplicates with technical duplicates. ( d ) BEA blocks the emergence of FLC resistance in C. albicans (SN95). 1×10 5 cells were plated on YEPD containing no inhibitor (−), 20 μg/ml BEA, 32 μg/ml FLC, or the combination. Plates were photographed after three days at 30°C. Experiment performed in biological triplicates.

    Journal: Nature chemical biology

    Article Title: Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling

    doi: 10.1038/nchembio.2165

    Figure Lengend Snippet: Beauvericin enhances fluconazole efficacy against diverse fungi and blocks the emergence of resistance ( a ) Beauvericin (BEA) reduces fluconazole resistance of reference strains of S. cerevisiae (BY4741), C. albicans (SN95), and C. neoformans (H99a), and A. fumigatus clinical isolate on rich medium (YEPD). Fluconazole (FLC) strips generate a gradient from 0.016 to 256 μg/ml, with the highest concentration at the top. Where indicated, plates contain 20 μg/ml of BEA. Experiment performed in biological triplicates. ( b ) BEA and FLC are synergistic and cidal against C. albicans (SN95). SN95 was subjected to two-fold serial dilutions of BEA and FLC in YEPD at 30°C for 48 hours. Optical densities were standardized to drug-free controls (see color bar) and FICI was calculated based on concentrations causing 60–70% growth inhibition. The drug combination is cidal, based on transferring cells to drug-free YEPD medium for 24 hours. ( c ) BEA reduced FLC resistance of C. albicans clinical isolates (CaCi) similar to geldanamycin (GdA) and cyclosporin A (CsA). CaCi are sequentially ordered with isolates recovered early at the top. MIC assays were performed in YEPD (−) with GdA (5 μM), CsA (20 μM), or BEA (25 μM), with a two-fold serial dilution of FLC. (b–c) Experiments performed in biological triplicates with technical duplicates. ( d ) BEA blocks the emergence of FLC resistance in C. albicans (SN95). 1×10 5 cells were plated on YEPD containing no inhibitor (−), 20 μg/ml BEA, 32 μg/ml FLC, or the combination. Plates were photographed after three days at 30°C. Experiment performed in biological triplicates.

    Article Snippet: Fluconazole (Sequoia Research Products) was dissolved in sterile ddH2 O, beauvericin in 100% methanol, cycloheximide (AG Scientific Inc.) in sterile ddH2 O, rhodamine-6G (Sigma-Aldrich) in sterile ddH2 O, geldanamycin (Invitrogen) in DMSO, and cyclosporin A (CalBiochem) in DMSO.

    Techniques: Concentration Assay, Inhibition, Transferring, Serial Dilution

    Diploid and Tetraploid C.albicans are sensitive to antifungals. A) Diploid colony forming units (CFUs) following 24-hr exposure to 1μg/mL or 10μg/mL fluconazole (‘FLU’, light and dark purple bars) and 0.25μg/mL or 2.5μg/mL caspofungin (‘CAS’, light and dark green bars) treatments. Bars represent the mean of 3 independent experiments (black symbols), and the error bars indicate +/− SEM. The dashed line and shaded box represent the mean and +/− SEM of the no-drug treatment. Asterisks indicate drug treatments that differ significantly from the no-drug treatment (* P

    Journal: G3: Genes|Genomes|Genetics

    Article Title: The Magnitude of Candida albicans Stress-Induced Genome Instability Results from an Interaction Between Ploidy and Antifungal Drugs

    doi: 10.1534/g3.119.400752

    Figure Lengend Snippet: Diploid and Tetraploid C.albicans are sensitive to antifungals. A) Diploid colony forming units (CFUs) following 24-hr exposure to 1μg/mL or 10μg/mL fluconazole (‘FLU’, light and dark purple bars) and 0.25μg/mL or 2.5μg/mL caspofungin (‘CAS’, light and dark green bars) treatments. Bars represent the mean of 3 independent experiments (black symbols), and the error bars indicate +/− SEM. The dashed line and shaded box represent the mean and +/− SEM of the no-drug treatment. Asterisks indicate drug treatments that differ significantly from the no-drug treatment (* P

    Article Snippet: Antifungal drug treatments were made from the following stock solutions: 1 mg fluconazole (ACROS Organics CAS#86386-73-4) was diluted into 1 mL DMSO, 1 mg caspofungin (Sigma-Aldrich CAS#179463-17-3) was diluted into 1 mL ddH2 0, and 10 mg calcofluor white (Sigma-Aldrich CAS#4404-43-7) was diluted into 1 mL ddH2 0.

    Techniques:

    Ploidy and antifungal drug-specific impacts on LOH in C. albicans . A) Diploid GAL1 LOH rates following 24-hr exposure to 1μg/mL or 10μg/mL fluconazole (‘FLU’, light and dark purple bars); 0.25μg/mL or 2.5μg/mL caspofungin (‘CAS’, light and dark green bars); and 100 μg/mL calcofluor white (’CW’, gray bar) treatments. Bars represent the mean of 3 independent experiments (black symbols), and the error bars indicate +/− SEM. The dashed line and shaded box represent the mean and +/− SEM of the no-drug treatment. Asterisks indicate drug treatments that differ significantly from the no-drug treatment (* P

    Journal: G3: Genes|Genomes|Genetics

    Article Title: The Magnitude of Candida albicans Stress-Induced Genome Instability Results from an Interaction Between Ploidy and Antifungal Drugs

    doi: 10.1534/g3.119.400752

    Figure Lengend Snippet: Ploidy and antifungal drug-specific impacts on LOH in C. albicans . A) Diploid GAL1 LOH rates following 24-hr exposure to 1μg/mL or 10μg/mL fluconazole (‘FLU’, light and dark purple bars); 0.25μg/mL or 2.5μg/mL caspofungin (‘CAS’, light and dark green bars); and 100 μg/mL calcofluor white (’CW’, gray bar) treatments. Bars represent the mean of 3 independent experiments (black symbols), and the error bars indicate +/− SEM. The dashed line and shaded box represent the mean and +/− SEM of the no-drug treatment. Asterisks indicate drug treatments that differ significantly from the no-drug treatment (* P

    Article Snippet: Antifungal drug treatments were made from the following stock solutions: 1 mg fluconazole (ACROS Organics CAS#86386-73-4) was diluted into 1 mL DMSO, 1 mg caspofungin (Sigma-Aldrich CAS#179463-17-3) was diluted into 1 mL ddH2 0, and 10 mg calcofluor white (Sigma-Aldrich CAS#4404-43-7) was diluted into 1 mL ddH2 0.

    Techniques:

    SDS-PAGE and phosphatase analysis of the mobility shift of hyperactive Tac1. (A) Immunoblot of Tac1 SDS-PAGE mobility in lysates from a 6His3Flag-tagged wild-type Tac1 strain (yLM485) treated with 10 μg/ml FNZ, 10 μg/ml EST, or 40 μg/ml fluconazole (FLC) for the indicated periods of time. Samples were resolved on 6% SDS-PAGE gel and the blot was probed with an anti-Flag antibody. Coomassie blue staining (CBS) was used as the loading control. (B) Immunoblot analysis of SDS-PAGE mobility in lysates from strains expressing an endogenous level of individual 6His3Flag-tagged Tac1 variants (wild-type [yLM485], N972D [yLM534], Δ962–969 [yLM535], ΔM677 [yLM533], R693K [yLM532], and A736V [yLM531]) in the absence and presence of 10 μg/ml fluphenazine. Samples were resolved on 6% SDS-PAGE gel and the blot was probed with an anti-Flag antibody. CBS was used as the loading control. (C) Immunoblot analysis demonstrating the effect of λ protein phosphatase (λ-pp) treatment on the SDS-PAGE mobility shift of hyperactive Tac1 purified from strains that overexpressed 6His3Flag (HF)-tagged wild-type Tac1 (yLM530; grown in the presence and absence of fluphenazine), Tac1 R693K (yLM540), or Tac1 N977D (yLM536). Samples were resolved by 6% SDS-PAGE and the blot was probed by an anti-Flag antibody. (D) Immunoblot analysis of the effect of λ protein phosphatase treatment (in the presence and absence of 50 mM NaF and 20 mM Na 3 VO 4 [F − /VO 4 3− ] phosphatase inhibitors) on the SDS-PAGE mobility shift of Tac1 purified from 6His3Flag-tagged wild-type Tac1 cells (yLM530) grown in the absence and presence of fluphenazine. Samples were resolved by 6% SDS-PAGE and the blot was probed by an anti-Flag antibody.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Mediator Tail Module Is Required for Tac1-Activated CDR1 Expression and Azole Resistance in Candida albicans

    doi: 10.1128/AAC.01342-17

    Figure Lengend Snippet: SDS-PAGE and phosphatase analysis of the mobility shift of hyperactive Tac1. (A) Immunoblot of Tac1 SDS-PAGE mobility in lysates from a 6His3Flag-tagged wild-type Tac1 strain (yLM485) treated with 10 μg/ml FNZ, 10 μg/ml EST, or 40 μg/ml fluconazole (FLC) for the indicated periods of time. Samples were resolved on 6% SDS-PAGE gel and the blot was probed with an anti-Flag antibody. Coomassie blue staining (CBS) was used as the loading control. (B) Immunoblot analysis of SDS-PAGE mobility in lysates from strains expressing an endogenous level of individual 6His3Flag-tagged Tac1 variants (wild-type [yLM485], N972D [yLM534], Δ962–969 [yLM535], ΔM677 [yLM533], R693K [yLM532], and A736V [yLM531]) in the absence and presence of 10 μg/ml fluphenazine. Samples were resolved on 6% SDS-PAGE gel and the blot was probed with an anti-Flag antibody. CBS was used as the loading control. (C) Immunoblot analysis demonstrating the effect of λ protein phosphatase (λ-pp) treatment on the SDS-PAGE mobility shift of hyperactive Tac1 purified from strains that overexpressed 6His3Flag (HF)-tagged wild-type Tac1 (yLM530; grown in the presence and absence of fluphenazine), Tac1 R693K (yLM540), or Tac1 N977D (yLM536). Samples were resolved by 6% SDS-PAGE and the blot was probed by an anti-Flag antibody. (D) Immunoblot analysis of the effect of λ protein phosphatase treatment (in the presence and absence of 50 mM NaF and 20 mM Na 3 VO 4 [F − /VO 4 3− ] phosphatase inhibitors) on the SDS-PAGE mobility shift of Tac1 purified from 6His3Flag-tagged wild-type Tac1 cells (yLM530) grown in the absence and presence of fluphenazine. Samples were resolved by 6% SDS-PAGE and the blot was probed by an anti-Flag antibody.

    Article Snippet: Fluconazole treatment was performed by adding 4 mg/ml fluconazole (Tokyo Chemical Industry Co.) in dimethyl sulfoxide (DMSO) stock to a mid-log-phase yLM485 culture to a final concentration of 40 μg/ml.

    Techniques: SDS Page, Mobility Shift, Staining, Expressing, Purification