Journal: PLoS ONE

Article Title: Cryptococcus neoformans Is Internalized by Receptor-Mediated or ‘Triggered’ Phagocytosis, Dependent on Actin Recruitment

doi: 10.1371/journal.pone.0089250

Figure Lengend Snippet: Quantification of the attachment to macrophages and the internalization of C. neoformans yeast cells from capsular (H99 and B3501) and acapsular (CAP67 and CAP 59) strains, in the presence of the actin polymerization inhibitors cytochalasin D or latrunculin B (A and B) or the microtubule stabilizers nocodazole or paclitaxel (C and D). Graphs show normalized mean values and standard deviation from three experiments. *p<0.05; **p<0.01; ***p<0.001.

Article Snippet: We used the following Cryptococcus neoformans strains: the capsular wild type strains H99 used for the genome project (ATCC 208821, serotype A,) and B3501 (serotype D); and the acapsular mutants CAP59 (NE367), derived from H99 , and CAP 67 (ATCC 52817), derived from B3501 .

Techniques: Standard Deviation

Journal: PLoS ONE

Article Title: Cryptococcus neoformans Is Internalized by Receptor-Mediated or ‘Triggered’ Phagocytosis, Dependent on Actin Recruitment

doi: 10.1371/journal.pone.0089250

Figure Lengend Snippet: Quantification of the internalization (A) and the attachment (B) to macrophages of C. neoformans yeast cells from capsular (H99 and B3501) and acapsular (CAP67 and CAP 59) strains, in the absence of cytoskeletal inhibitors or in the presence of cytochalasin D and nocodazole. The metabolic viability of C. neoformans strains H99 and CAP59 was measured using the FUN®-1 dye (C) and the metabolic viability of macrophages was measured by MTS/PMS (D) after incubation with cytoskeletal inhibitors for 2 h. Yeast cells fixed with 70% ethanol, and macrophages with 4% formaldehyde, were used as a positive control for the loss of cell viability in each method. Graphs show normalized mean values and standard deviation from three experiments (A–B) and mean and standard deviation from absolute values of fluorescence intensity (C) and absorbance (D).*p<0.05; **p<0.01; ***p<0.001.

Article Snippet: We used the following Cryptococcus neoformans strains: the capsular wild type strains H99 used for the genome project (ATCC 208821, serotype A,) and B3501 (serotype D); and the acapsular mutants CAP59 (NE367), derived from H99 , and CAP 67 (ATCC 52817), derived from B3501 .

Techniques: Incubation, Positive Control, Standard Deviation, Fluorescence

Journal: PLoS ONE

Article Title: Cryptococcus neoformans Is Internalized by Receptor-Mediated or ‘Triggered’ Phagocytosis, Dependent on Actin Recruitment

doi: 10.1371/journal.pone.0089250

Figure Lengend Snippet: Scanning electron microscopy of membrane extracted macrophages interacting with C. neoformans strains H99 (A) and CAP59 (B and C), showing cytoskeletal filaments associated with yeasts in untreated samples (A–B). After 5 µm nocodazole treatment (C) the area surrounding yeast cells appeared mostly devoid of cytoskeletal components but association with yeast still occurred (inset in C). Scale bars, 2 µm.

Article Snippet: We used the following Cryptococcus neoformans strains: the capsular wild type strains H99 used for the genome project (ATCC 208821, serotype A,) and B3501 (serotype D); and the acapsular mutants CAP59 (NE367), derived from H99 , and CAP 67 (ATCC 52817), derived from B3501 .

Techniques: Electron Microscopy, Membrane

Journal: PLoS ONE

Article Title: Cryptococcus neoformans Is Internalized by Receptor-Mediated or ‘Triggered’ Phagocytosis, Dependent on Actin Recruitment

doi: 10.1371/journal.pone.0089250

Figure Lengend Snippet: Confocal laser scanning microscopy (z-stack series of confocal planes) of interacting macrophages and C. neoformans yeast cells from strains H99 (A and B) and CAP59 (C and D). Internalized yeasts identified by DIC (arrows in A and C) can be visualized in the context of host cell actin (red) and microtubule (green) cytoskeletons (B and D). Host cell DNA is labeled with DAPI (blue, indicated by the letter ‘N’) and yeast is labeled with calcofluor (blue, indicated by arrows). Actin, but not tubulin, is recruited to sites of yeast internalization. Scale bars, 5 µm.

Article Snippet: We used the following Cryptococcus neoformans strains: the capsular wild type strains H99 used for the genome project (ATCC 208821, serotype A,) and B3501 (serotype D); and the acapsular mutants CAP59 (NE367), derived from H99 , and CAP 67 (ATCC 52817), derived from B3501 .

Techniques: Confocal Laser Scanning Microscopy, Labeling

Journal: PLoS ONE

Article Title: Cryptococcus neoformans Is Internalized by Receptor-Mediated or ‘Triggered’ Phagocytosis, Dependent on Actin Recruitment

doi: 10.1371/journal.pone.0089250

Figure Lengend Snippet: Confocal laser scanning microscopy of C. neoformans capsular strain H99 interacting with macrophages (single confocal plane). DIC showing internalized yeasts (arrows); and confocal images showing actin filaments (red), microtubules (green), yeast (blue) and host DNA (blue, indicated by ‘n’). Actin is recruited to the site of phagocytosis in untreated cells (A), and actin recruitment was inhibited by 0.5 µM cytochalasin D (B). In contrast, treatment with 5 µM nocodazole (C) or with a combination of nocodazole and cytochalasin D (D) did not inhibit actin recruitment to the phagosome area. Scale bars, 5 µm.

Article Snippet: We used the following Cryptococcus neoformans strains: the capsular wild type strains H99 used for the genome project (ATCC 208821, serotype A,) and B3501 (serotype D); and the acapsular mutants CAP59 (NE367), derived from H99 , and CAP 67 (ATCC 52817), derived from B3501 .

Techniques: Confocal Laser Scanning Microscopy

Journal: PLoS ONE

Article Title: Cryptococcus neoformans Is Internalized by Receptor-Mediated or ‘Triggered’ Phagocytosis, Dependent on Actin Recruitment

doi: 10.1371/journal.pone.0089250

Figure Lengend Snippet: Scanning electron microscopy of C. neoformans capsular strain H99 (A–E) and acapsular strain CAP59 (F–G) interacting with peritoneal macrophages. Improved preservation of macrophage membranes was obtained with routine SEM fixation (A–B; F–G), although post-fixation in the presence of sucrose provided better capsule preservation and allowed visualization of direct interactions between the capsule and host cell membranes, prior to internalization (C–E). Both trigger-like (arrow in A and F) and zipper-like (arrow-head in B and G) uptake structures were observed. Scale bars, 1 µm (A–C; F–G) and 0.5 µm (D–E).

Article Snippet: We used the following Cryptococcus neoformans strains: the capsular wild type strains H99 used for the genome project (ATCC 208821, serotype A,) and B3501 (serotype D); and the acapsular mutants CAP59 (NE367), derived from H99 , and CAP 67 (ATCC 52817), derived from B3501 .

Techniques: Electron Microscopy, Preserving

Journal: PLoS ONE

Article Title: Cryptococcus neoformans Is Internalized by Receptor-Mediated or ‘Triggered’ Phagocytosis, Dependent on Actin Recruitment

doi: 10.1371/journal.pone.0089250

Figure Lengend Snippet: Quantification of the uptake mechanisms during C. neoformans -macrophage interaction.

Article Snippet: We used the following Cryptococcus neoformans strains: the capsular wild type strains H99 used for the genome project (ATCC 208821, serotype A,) and B3501 (serotype D); and the acapsular mutants CAP59 (NE367), derived from H99 , and CAP 67 (ATCC 52817), derived from B3501 .

Techniques:

Journal: Applied and Environmental Microbiology

Article Title: Toxoflavin Produced by Burkholderia gladioli from Lycoris aurea Is a New Broad-Spectrum Fungicide

doi: 10.1128/AEM.00106-19

Figure Lengend Snippet: Structural analysis and characterization of the main active antimicrobial ingredient produced by strain HDXY-02. (A) HPLC analysis of extraction separated from strain HDXY-02 culture broth. (B) Identification of toxoflavin with MALDI-Q-TOF-MS. (C) TLC analysis under UV light at 365 nm. (D) Antimicrobial assay. Serial dilutions of 1 μl of 10-fold conidia (107 to 105) of A. fumigatus, A. nidulans, C. albicans, and C. neoformans were spotted onto solid medium containing various toxoflavin concentrations (0 to 200 μg/ml).

Article Snippet: These data suggest that toxoflavin might have a fungal-inhibition mechanism different from that of traditional azole antifungals. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Species Strain Concn (μg/ml) MIC MFC M. oryzae HN13-2-1-1 128 128 R. solani RS01 128 256 F. graminearum 2021 256 >256 A. fumigatus Af293 (FGSC A1100) 64 128 A. fumigatus FGSC A1160 64 128 A. fumigatus AfCox10 243Q (WX01) 64 128 A. fumigatus AfCyp51A F219L (WX03) 64 128 A. nidulans LB01 (FGSC A118) 128 256 C. albicans ATCC 10231 64 256 C. neoformans H99 (ATCC 208821) 128 128 Open in a separate window MICs and MFCs of toxoflavin isolated from the culture filtrate of strain HDXY-02 against several fungi To characterize the effects of toxoflavin on A. fumigatus Af293, a microscopy study was carried out.

Techniques: Produced, Extraction

Journal: Applied and Environmental Microbiology

Article Title: Toxoflavin Produced by Burkholderia gladioli from Lycoris aurea Is a New Broad-Spectrum Fungicide

doi: 10.1128/AEM.00106-19

Figure Lengend Snippet: MICs and MFCs of toxoflavin isolated from the culture filtrate of strain HDXY-02 against several fungi

Article Snippet: These data suggest that toxoflavin might have a fungal-inhibition mechanism different from that of traditional azole antifungals. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Species Strain Concn (μg/ml) MIC MFC M. oryzae HN13-2-1-1 128 128 R. solani RS01 128 256 F. graminearum 2021 256 >256 A. fumigatus Af293 (FGSC A1100) 64 128 A. fumigatus FGSC A1160 64 128 A. fumigatus AfCox10 243Q (WX01) 64 128 A. fumigatus AfCyp51A F219L (WX03) 64 128 A. nidulans LB01 (FGSC A118) 128 256 C. albicans ATCC 10231 64 256 C. neoformans H99 (ATCC 208821) 128 128 Open in a separate window MICs and MFCs of toxoflavin isolated from the culture filtrate of strain HDXY-02 against several fungi To characterize the effects of toxoflavin on A. fumigatus Af293, a microscopy study was carried out.

Techniques: Isolation