Journal: PLoS ONE

Article Title: Chromosomal Polymorphism in the Sporothrix schenckii Complex

doi: 10.1371/journal.pone.0086819

Figure Lengend Snippet: Strains, species, origin, and GenBank accession numbers (CAL and ITS) for the Sporothrix spp. isolates used in this study.

Article Snippet: Based on genomic data from the genome sequencing project of S. schenckii s. str. (ATCC MYA-4821) and S. brasiliensis (ATCC MYA-4823) isolates, the genome sizes of ATCC MYA-4821 and ATCC MYA-4823 were estimated to be 32.5 Mb and 33.4 Mb, respectively (Marcus de Melo Teixeira and Maria Sueli Soares Felipe, personal communication).

Techniques:

Journal: PLoS ONE

Article Title: Chromosomal Polymorphism in the Sporothrix schenckii Complex

doi: 10.1371/journal.pone.0086819

Figure Lengend Snippet: Each species is grouped inside a colored rectangle. Red: S. schenckii ; Yellow: S. brasiliensis ; Blue: S. mexicana ; Purple: S. luriei . The sizes of chromosomal bands (Mb) are shown on the left. Σ: The minimum genome sizes estimated by summing the chromosomal bands (Mb) are shown at the bottom.

Article Snippet: Based on genomic data from the genome sequencing project of S. schenckii s. str. (ATCC MYA-4821) and S. brasiliensis (ATCC MYA-4823) isolates, the genome sizes of ATCC MYA-4821 and ATCC MYA-4823 were estimated to be 32.5 Mb and 33.4 Mb, respectively (Marcus de Melo Teixeira and Maria Sueli Soares Felipe, personal communication).

Techniques:

Journal: PLoS ONE

Article Title: Chromosomal Polymorphism in the Sporothrix schenckii Complex

doi: 10.1371/journal.pone.0086819

Figure Lengend Snippet: The S. schenckii and S. brasiliensis isolates are indicated at the top. Each grey box represents a chromosomal band with the indicated size (Mb, left ). The probes that hybridized to each band are color-coded ( bottom ). β-tubulin (β-Tub), calmodulin (Calm), catalase (Cata), chitin synthase 1 (CHS1), internal transcribed spacer (ITS), Pho85 cyclin-dependent kinase (Pho85), protein kinase C Ss-2 (PKC), G protein α subunit (GProt), and topoisomerase II (Topo).

Article Snippet: Based on genomic data from the genome sequencing project of S. schenckii s. str. (ATCC MYA-4821) and S. brasiliensis (ATCC MYA-4823) isolates, the genome sizes of ATCC MYA-4821 and ATCC MYA-4823 were estimated to be 32.5 Mb and 33.4 Mb, respectively (Marcus de Melo Teixeira and Maria Sueli Soares Felipe, personal communication).

Techniques:

Journal: PLoS ONE

Article Title: Chromosomal Polymorphism in the Sporothrix schenckii Complex

doi: 10.1371/journal.pone.0086819

Figure Lengend Snippet: A: The average minimum genome size for different Sporothrix species was estimated by summing all the chromosomal bands detected after pulsed field gel electrophoresis. B: Comparison of genome sizes estimated from genome sequencing (blue) and estimated from the sum of chromosomal bands (red) for isolates ATCC MYA-4821 ( S. schenckii ) and ATCC MYA-4823 ( S. brasiliensis ); the differences in size estimated with the different methodologies are indicated on the right.

Article Snippet: Based on genomic data from the genome sequencing project of S. schenckii s. str. (ATCC MYA-4821) and S. brasiliensis (ATCC MYA-4823) isolates, the genome sizes of ATCC MYA-4821 and ATCC MYA-4823 were estimated to be 32.5 Mb and 33.4 Mb, respectively (Marcus de Melo Teixeira and Maria Sueli Soares Felipe, personal communication).

Techniques: Pulsed-Field Gel, Electrophoresis, Sequencing

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Growth inhibition of Sporothrix brasiliensis yeasts after exposure to the 400 compounds of the Pandemic Response Box library for 48h. Itraconazole (in bold) is the antifungal recommended for sporotrichosis treatment. Results are shown as the mean of percent growth relative to the untreated control.

Article Snippet: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/jof8101004/s1 , Table S1: Inhibition of Sporothrix brasiliensis ATCC MYA 4823 in yeast phase after exposure to 1 μM of the Pandemic Response Box compounds.

Techniques: Inhibition

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Inhibitory effects of twenty-four compounds on the pathogenic Sporothrix species.

Article Snippet: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/jof8101004/s1 , Table S1: Inhibition of Sporothrix brasiliensis ATCC MYA 4823 in yeast phase after exposure to 1 μM of the Pandemic Response Box compounds.

Techniques: Concentration Assay

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: The anti- Sporothrix activity of olorofim. The chemical structure of olorofim ( A ). After exposure to olorofim or itraconazole, the yeast killing was evaluated by cultivation in a drug-free medium ( B ). The ability of olorofim to inhibit mature biofilms was determined according to the metabolic activity of cells in the presence of compounds ( C ). Results are shown as the mean of percent growth or metabolic activity and the standard error of the mean.

Article Snippet: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/jof8101004/s1 , Table S1: Inhibition of Sporothrix brasiliensis ATCC MYA 4823 in yeast phase after exposure to 1 μM of the Pandemic Response Box compounds.

Techniques: Activity Assay

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Cytotoxicities of olorofim and itraconazole.

Article Snippet: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/jof8101004/s1 , Table S1: Inhibition of Sporothrix brasiliensis ATCC MYA 4823 in yeast phase after exposure to 1 μM of the Pandemic Response Box compounds.

Techniques:

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Effects of olorofim exposure on S. brasiliensis yeasts. Treated yeasts analyzed by fluorimetry exhibited a dose−dependent DNA accumulation ( A ), increased chitin ( B ), decreased β−glucan ( C ), increased mannan ( D ), and reduced electronegativity ( E ) and conductance ( F ). Cell wall measurements revealed that olorofim treatment increased the thickness of this structure ( G ). Scanning electron microscopy images showed untreated yeasts with an elongated shape ( Hi ), and treated cells with alterations in cell wall integrity, exhibiting a granular appearance (arrow in Hii ). Transmission electron microscopy images illustrated untreated cells with homogeneous cytoplasm with nucleus (n) and mitochondria (m), surrounded by a plasma membrane and cell wall (cw) ( Hiii ), while treatment with olorofim induced an amorphous shape and modifications in the cell wall structure (arrow in Hiv ). Graphs showed the min and maximum with the line in the median, with the X −axis representing olorofim concentrations in µM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant by one-way ANOVA. Scale bars: 2.5 µm ( Hi , Hii ) and 0.5 µm ( Hiii , Hiv ).

Article Snippet: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/jof8101004/s1 , Table S1: Inhibition of Sporothrix brasiliensis ATCC MYA 4823 in yeast phase after exposure to 1 μM of the Pandemic Response Box compounds.

Techniques: Electron Microscopy, Transmission Assay

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Interaction between keratinocytes and S. brasiliensis yeasts. Yeasts treated with MIC of compounds (0.06 µM olorofim or 0.125 µM itraconazole) displayed decreased ability to adhere to the surface of keratinocytes. * p < 0.05 and ** p < 0.01 by one-way ANOVA.

Article Snippet: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/jof8101004/s1 , Table S1: Inhibition of Sporothrix brasiliensis ATCC MYA 4823 in yeast phase after exposure to 1 μM of the Pandemic Response Box compounds.

Techniques:

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Schematic model of olorofim action against Sporothrix yeasts. Olorofim belongs to a new class of antifungal agents called the orotomides, which inhibits fungal pyrimidine biosynthesis. Olorofim inhibits dihydroorotate dehydrogenase, an enzyme of de novo pyrimidine biosynthesis, leading to the rapid depletion of uridine 5′-monophosphate (UMP), uridine diphosphate (UDP), and uridine triphosphate (UTP). The reduction in UTP levels can cause a decrease in cell wall biogenesis, protein glycosylation, and synthesis of DNA, protein, and phospholipid. In Sporothrix yeast cells, the inhibition of pyrimidine synthesis by olorofim may induce cell cycle arrest and affect the fungal cell wall, resulting in cell lysis. The illustration was partially based on Servier Medical Art elements and licensed under a Creative Commons Attribution 3.0 Unported License. ER: endoplasmic reticulum.

Article Snippet: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/jof8101004/s1 , Table S1: Inhibition of Sporothrix brasiliensis ATCC MYA 4823 in yeast phase after exposure to 1 μM of the Pandemic Response Box compounds.

Techniques: Inhibition, Lysis

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Growth inhibition of Sporothrix brasiliensis yeasts after exposure to the 400 compounds of the Pandemic Response Box library for 48h. Itraconazole (in bold) is the antifungal recommended for sporotrichosis treatment. Results are shown as the mean of percent growth relative to the untreated control.

Article Snippet: The reference isolates S. brasiliensis ATCC MYA 4823, S. schenckii ATCC 32286, and S. globosa CBS 130104 were used in this study.

Techniques: Inhibition

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Inhibitory effects of twenty-four compounds on the pathogenic Sporothrix species.

Article Snippet: The reference isolates S. brasiliensis ATCC MYA 4823, S. schenckii ATCC 32286, and S. globosa CBS 130104 were used in this study.

Techniques: Concentration Assay

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Effects of olorofim exposure on S. brasiliensis yeasts. Treated yeasts analyzed by fluorimetry exhibited a dose−dependent DNA accumulation ( A ), increased chitin ( B ), decreased β−glucan ( C ), increased mannan ( D ), and reduced electronegativity ( E ) and conductance ( F ). Cell wall measurements revealed that olorofim treatment increased the thickness of this structure ( G ). Scanning electron microscopy images showed untreated yeasts with an elongated shape ( Hi ), and treated cells with alterations in cell wall integrity, exhibiting a granular appearance (arrow in Hii ). Transmission electron microscopy images illustrated untreated cells with homogeneous cytoplasm with nucleus (n) and mitochondria (m), surrounded by a plasma membrane and cell wall (cw) ( Hiii ), while treatment with olorofim induced an amorphous shape and modifications in the cell wall structure (arrow in Hiv ). Graphs showed the min and maximum with the line in the median, with the X −axis representing olorofim concentrations in µM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant by one-way ANOVA. Scale bars: 2.5 µm ( Hi , Hii ) and 0.5 µm ( Hiii , Hiv ).

Article Snippet: The reference isolates S. brasiliensis ATCC MYA 4823, S. schenckii ATCC 32286, and S. globosa CBS 130104 were used in this study.

Techniques: Electron Microscopy, Transmission Assay

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Interaction between keratinocytes and S. brasiliensis yeasts. Yeasts treated with MIC of compounds (0.06 µM olorofim or 0.125 µM itraconazole) displayed decreased ability to adhere to the surface of keratinocytes. * p < 0.05 and ** p < 0.01 by one-way ANOVA.

Article Snippet: The reference isolates S. brasiliensis ATCC MYA 4823, S. schenckii ATCC 32286, and S. globosa CBS 130104 were used in this study.

Techniques:

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Growth inhibition of Sporothrix brasiliensis yeasts after exposure to the 400 compounds of the Pandemic Response Box library for 48h. Itraconazole (in bold) is the antifungal recommended for sporotrichosis treatment. Results are shown as the mean of percent growth relative to the untreated control.

Article Snippet: The reference isolate S. brasiliensis ATCC MYA 4823 was used to screen the Pandemic Response Box compounds for inhibitors of fungal growth.

Techniques: Inhibition

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Inhibitory effects of twenty-four compounds on the pathogenic Sporothrix species.

Article Snippet: The reference isolate S. brasiliensis ATCC MYA 4823 was used to screen the Pandemic Response Box compounds for inhibitors of fungal growth.

Techniques: Concentration Assay

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Effects of olorofim exposure on S. brasiliensis yeasts. Treated yeasts analyzed by fluorimetry exhibited a dose−dependent DNA accumulation ( A ), increased chitin ( B ), decreased β−glucan ( C ), increased mannan ( D ), and reduced electronegativity ( E ) and conductance ( F ). Cell wall measurements revealed that olorofim treatment increased the thickness of this structure ( G ). Scanning electron microscopy images showed untreated yeasts with an elongated shape ( Hi ), and treated cells with alterations in cell wall integrity, exhibiting a granular appearance (arrow in Hii ). Transmission electron microscopy images illustrated untreated cells with homogeneous cytoplasm with nucleus (n) and mitochondria (m), surrounded by a plasma membrane and cell wall (cw) ( Hiii ), while treatment with olorofim induced an amorphous shape and modifications in the cell wall structure (arrow in Hiv ). Graphs showed the min and maximum with the line in the median, with the X −axis representing olorofim concentrations in µM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant by one-way ANOVA. Scale bars: 2.5 µm ( Hi , Hii ) and 0.5 µm ( Hiii , Hiv ).

Article Snippet: The reference isolate S. brasiliensis ATCC MYA 4823 was used to screen the Pandemic Response Box compounds for inhibitors of fungal growth.

Techniques: Electron Microscopy, Transmission Assay

Journal: Journal of Fungi

Article Title: Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species

doi: 10.3390/jof8101004

Figure Lengend Snippet: Interaction between keratinocytes and S. brasiliensis yeasts. Yeasts treated with MIC of compounds (0.06 µM olorofim or 0.125 µM itraconazole) displayed decreased ability to adhere to the surface of keratinocytes. * p < 0.05 and ** p < 0.01 by one-way ANOVA.

Article Snippet: The reference isolate S. brasiliensis ATCC MYA 4823 was used to screen the Pandemic Response Box compounds for inhibitors of fungal growth.

Techniques: