minimal inhibitory concentration mic determination  (ATCC)

Journal: Antimicrobial Agents and Chemotherapy

Article Title: The siderophore transporter Sit1 is involved in the uptake of caspofungin by Candida albicans

doi: 10.1128/aac.01236-25

Figure Lengend Snippet: Functional enrichment of C. albicans gene expression after treatment with iron, caspofungin, or both. Enrichment analysis of the differentially expressed genes. Only enriched categories (FDR < 0.05 and fold enrichment > 2) are represented. The size and color of the circles represent the number of genes associated with each term and FDR, respectively.

Article Snippet: Cell suspensions were evenly spread onto SC agar plates, and caspofungin minimal inhibitory concentration (MIC) Test Strips (Liofilchem) were placed on the center of the plate using sterile tweezers.

Techniques: Functional Assay, Gene Expression

Journal: Antimicrobial Agents and Chemotherapy

Article Title: The siderophore transporter Sit1 is involved in the uptake of caspofungin by Candida albicans

doi: 10.1128/aac.01236-25

Figure Lengend Snippet: Caspofungin affects iron homeostasis. ( A ) Heatmap depicting C. albicans genes related to the subcategory of iron homeostasis, which were differentially expressed under Fe, CAS, or Fe + CAS conditions. For each condition, the log 2 fold change (log 2 FC) of the selected transcripts is indicated using a color code. Genes significantly upregulated (log 2 FC > 1) or downregulated (log 2 FC < –1) in response to Fe, CAS, or Fe + CAS (treated versus untreated conditions) are shaded in red or blue, respectively. Asterisks (*) mark genes differentially expressed with FDR < 0.3 and/or log 2 CPM > 1.3 and therefore do not meet the more stringent criteria of log₂FC > 1, log 2 CPM > 3, and FDR < 0.05 and are not listed in . The complete data set is available at NCBI GEO under accession number GSE280500 . S. cerevisiae gene names were used whenever a C. albicans gene name was not assigned. ( B ) The iron content of C. albicans SC5314 cells left untreated (Control) or treated overnight with Fe, CAS, or both (Fe + CAS) was determined by ICP-AES. Significance of differences was calculated using one-way ANOVA with Tukey’s HSD post hoc test (**** P < 0.0001; n.s. not significant). ( C ) Growth of S. cerevisiae wild type (wt, BY4742), Δ ccc1 , Δ fks1, and Δ fks1ccc1 strains on SC agar plates (Control) containing Fe (5 mM), CAS (0.03 µg/mL), or Fe + CAS, for 3 days at 30°C.

Article Snippet: Cell suspensions were evenly spread onto SC agar plates, and caspofungin minimal inhibitory concentration (MIC) Test Strips (Liofilchem) were placed on the center of the plate using sterile tweezers.

Techniques: Control

Journal: Antimicrobial Agents and Chemotherapy

Article Title: The siderophore transporter Sit1 is involved in the uptake of caspofungin by Candida albicans

doi: 10.1128/aac.01236-25

Figure Lengend Snippet: Sit1 affects caspofungin efficacy against yeast. ( A ) Structural similarity between caspofungin and cyclic hexapeptides hydroxamate siderophores. ( B ) Growth of C. albicans wt (CAF2-1) and ΔΔ sit1 mutant strains on SC agar plates (Control) containing 0.4 µM caspofungin (CAS), for 48 h or 5 days (CAS *) at 30°C. Growth of S. cerevisiae wt (YPH499) and Δ arn1-4 mutant strains on SC agar plates (Control) containing 0.1 µM caspofungin (CAS), for 48 h at 30°C. ( C ) C. albicans cells were left untreated (Control) or treated with 0.375 µg/mL caspofungin (CAS) for 3 h and plated on YPD agar plates for CFU count. Significance of differences was calculated using two-way ANOVA with Tukey’s HSD post hoc test (*** P < 0.001; * P < 0.05). ( D ) Growth of S. cerevisiae wt (YPH499) and Δ arn1-4 mutant strains transformed with a plasmid containing the CaSIT1 gene under the control of ScPGK1 promoter (p PGK-SIT1 ) or with the empty vector (v) on SC-ura agar plates (Control) containing the indicated concentrations of caspofungin (CAS), for 48 h at 30°C.

Article Snippet: Cell suspensions were evenly spread onto SC agar plates, and caspofungin minimal inhibitory concentration (MIC) Test Strips (Liofilchem) were placed on the center of the plate using sterile tweezers.

Techniques: Mutagenesis, Control, Transformation Assay, Plasmid Preparation

Journal: Antimicrobial Agents and Chemotherapy

Article Title: The siderophore transporter Sit1 is involved in the uptake of caspofungin by Candida albicans

doi: 10.1128/aac.01236-25

Figure Lengend Snippet: Synthesis of fluorescent FAM-labeled caspofungin (CAS-FAM) (1). Reagents and conditions: (a) Boc 2 O, dioxane/H 2 O (1:1), room temperature, 48 h, 89%; (b) propargyl bromide, Cs 2 CO 3 , DMF, room temperature, 14 h, 67%; (c) fluorescein azide, 2,2′-bipyridine, CuI, sodium ascorbate, DMF, room temperature, 4 h, 77%; (d) 37% HCl, H 2 O/isopropyl alcohol (1:3, vol/vol), 2 h, room temperature, 85%.

Article Snippet: Cell suspensions were evenly spread onto SC agar plates, and caspofungin minimal inhibitory concentration (MIC) Test Strips (Liofilchem) were placed on the center of the plate using sterile tweezers.

Techniques: Labeling

Journal: Antimicrobial Agents and Chemotherapy

Article Title: The siderophore transporter Sit1 is involved in the uptake of caspofungin by Candida albicans

doi: 10.1128/aac.01236-25

Figure Lengend Snippet: Sit1 mediates the uptake of CAS-FAM. ( A ) Growth of caspofungin-sensitive ( C. albicans SC5314, laboratory strain) and resistant strains ( Ca BS1, Ca BS2, and Ca 13-514) on SC agar plates containing 0.4 µM caspofungin (CAS) or 4 µM CAS-FAM, after 24 h at 30°C. ( B ) Growth of S. cerevisiae wt (YPH499) and Δ arn1-4 mutant strains transformed with a plasmid containing the CaSIT1 gene under the control of ScPGK1 promoter (p PGK-SIT1 ) or with the empty vector (v) on SC-ura agar plates (Control) containing the indicated concentrations of CAS-FAM, after 48 h at 30°C. ( C ) Quantification of the intracellular fluorescence levels of S. cerevisiae cells left untreated (Control) or treated with 1 µM CAS-FAM for 1 h. Significance of differences was calculated using two-way ANOVA with Tukey’s HSD post hoc test (**** P < 0.0001; ** P < 0.01). ( D ) Quantification of the intracellular fluorescence levels of S. cerevisiae Δ arn1-4 cells transformed with p PGK- SIT1 left untreated (Control) or treated with 1 µM CAS-FAM (CAS-FAM), 500 µM FeSO 4 (Fe), or both (Fe + CAS FAM) for 1 h was measured. Significance of differences was calculated using one-way ANOVA with Tukey’s HSD post hoc test (**** P < 0.0001; ** P < 0.01). ( E ) Fluorescence microscopy images of S. cerevisiae wt cells transformed with p PGK- SIT1 treated with 1.5 µM CAS-FAM (+ CAS FAM) for 1.5 h. Scale bar: 5 μm; arrow heads: vacuoles, BF: bright field. ( F ) Quantification of fluorescence microscopy images of cells treated with 1.5 µM CAS-FAM (CAS-FAM) for 1.5 h, either alone or in combination with 10 µM ferrichrome (CAS-FAM + FC). At least 100 cells were analyzed per condition. Significance of differences was calculated using Student’s T-test (**** P < 0.0001).

Article Snippet: Cell suspensions were evenly spread onto SC agar plates, and caspofungin minimal inhibitory concentration (MIC) Test Strips (Liofilchem) were placed on the center of the plate using sterile tweezers.

Techniques: Mutagenesis, Transformation Assay, Plasmid Preparation, Control, Fluorescence, Microscopy

Journal: Antimicrobial Agents and Chemotherapy

Article Title: The siderophore transporter Sit1 is involved in the uptake of caspofungin by Candida albicans

doi: 10.1128/aac.01236-25

Figure Lengend Snippet: Sit1 is involved in the uptake of caspofungin in C. albicans . ( A ) The accumulation of caspofungin in C. albicans wt (CAF2-1) and ΔΔ sit1 cells grown in iron-depleted medium (supplemented with 300 µM of the iron chelator BPS) or iron-replete medium and treated with 1 µg/mL caspofungin for 2 h was measured by LC-MS (**** P < 0.0001; *** P < 0.001). ( B ) The iron content of C. albicans wt (CAF2-1) and ΔΔ sit1 cells left untreated (Control) or treated overnight with 5 mM FeSO 4 (Fe), 0.375 µg/mL caspofungin (CAS), or both (Fe + CAS) was determined by ICP-AES. Significance of differences was calculated using two-way ANOVA with Tukey’s HSD post hoc test (**** P < 0.0001).

Article Snippet: Cell suspensions were evenly spread onto SC agar plates, and caspofungin minimal inhibitory concentration (MIC) Test Strips (Liofilchem) were placed on the center of the plate using sterile tweezers.

Techniques: Liquid Chromatography with Mass Spectroscopy, Control