grubii serotype a  (ATCC)

Journal: bioRxiv

Article Title: Spatiotemporal dynamics of cryptococcal infection reveal novel immune modulatory mechanisms and antifungal targets

doi: 10.1101/2025.05.30.657021

Figure Lengend Snippet: A. Capsule production of C. neoformans H99 and 05069 Δ strains in capsule-inducing media (i.e., LIM) measured by India ink staining and differential interference microscopy. Scale bar = 1.0 µm. Capsule thickness and cell size quantification for H99 and 05069Δ strains from a minimum of 50 cells. Statistical significance assessed using Student’s t -test: * P < 0.05, *** P < 0.0001. Ratio of capsule thickness to cell diameter calculated. B. Percentage of melanin pigmentation for C. neoformans strains from the L-DOPA plate assay. Values normalized to H99 at 30 °C (left) and 37 °C (right). C. Fungal growth of C. neoformans H99 and 05069 Δ strains on plates of YNB with amino acids supplemented with 0.05% dextrose at 30 °C and 37 °C and grown in YNB with amino acids supplemented with 0.05% dextrose liquid media. Fungal growth measured at OD 600nm . AUC quantification of C. neoformans H99 and 05069 Δ strains growth in YNB at 30 °C and 37 °C. D. Host cell cytotoxicity assay by quantification of LDH release upon macrophage cell death following coculture with C. neoformans H99 and 05069 Δ. Uninfected control refers to macrophage-only culture. AUC quantification of C. neoformans WT and 05069 Δ strains, and uninfected host cell death from LDH time course assay over 48 h. Two independent biological mutant strains constructed; experiments performed in biological triplicate and technical duplicate. E. and F. Superimposition of predicted structures for C. neoformans cQCR9, cQCR10 (CNAG_05069; an ubiquinol-cytochrome c reductase subunit 10), and cRip1. Subunits 9, 10 and Rip1 from S. cerevisiae = green, red and blue, respectively. Subunits 9, 10 and Rip1 from C. neoformans = salmon, yellow and pink, respectively. G. Interaction between cQCR9 and cQCR10 amino acids; colored based on hydrophobicity. H. Altered A36W and V29W with shortening of peptide length. Original position of cQCR10 (white) highlights new orientation of cQCR10-V29W-A36W-short. Amino acid labels from cQCR10 = black, cQCR9 = blue. *Amino acids selected for mutation analysis. Proteins displayed in cartoon mode. Intermembrane space (IMS), inner membrane (IM) and matrix regions are displayed. iPTM = interface predicted template modeling; LIS = local interaction score. Figures prepared in Pymol 3.1.4.1 ( https://pymol.org/ ).

Article Snippet: A comprehensive spectral library was generated using protein FASTA files from C. neoformans var. grubii serotype A (strain H99/ATCC 208821, UniProt UP000010091; 7,429 sequences) and Mus musculus (17,836 Swiss-Prot reviewed entries, UniProt) .

Techniques: Staining, Microscopy, Cytotoxicity Assay, Control, Mutagenesis, Construct, Membrane

Journal: bioRxiv

Article Title: Spatiotemporal dynamics of cryptococcal infection reveal novel immune modulatory mechanisms and antifungal targets

doi: 10.1101/2025.05.30.657021

Figure Lengend Snippet: A. Volcano plot of significantly different protein identified from infected vs. uninfected brain tissue independent of time (i.e., 3 dpi, 10 dpi, endpoint). Student’s t-test p-value < 0.05, FDR = 5%, S 0 = 1. B. Normalized LFQ intensity of haptoglobin from infected and uninfected temporal samples from murine brain, lungs, and spleen. Statistical significanc assessed using Student’s t -test: * P < 0.05, ** P < 0.01, *** P < 0.0001. C. Western blot (and corresponding Coomassie SDS-PAGE) for haptoglobin produced in infected and uninfected brain, lungs, and spleen murine samples. L = ladder, 3 U/I = 3 dpi uninfected or infected, 10 U/I = 10 dpi uninfected or infected, EU/I = endpoint uninfected or infection. Haptoglobin = 39 kDa. Semin-quantification provided. Normalized to background for each blot. D. CFU quantification of C. neoformans following co-culture with macrophage (BALB/c, Hp-siRNA). Percent survival of fungi depicted relative to initial inoculum concentration. E. Host cell cytotoxicity assay by quantification of LDH release upon macrophage (BALB/c, Hp-siRNA) cell death following coculture with C. neoformans H99. Two independent biological mutant strains constructed; experiments performed in biological triplicate and technical duplicate.

Article Snippet: A comprehensive spectral library was generated using protein FASTA files from C. neoformans var. grubii serotype A (strain H99/ATCC 208821, UniProt UP000010091; 7,429 sequences) and Mus musculus (17,836 Swiss-Prot reviewed entries, UniProt) .

Techniques: Infection, Western Blot, SDS Page, Produced, Co-Culture Assay, Concentration Assay, Cytotoxicity Assay, Mutagenesis, Construct

Journal: Antioxidants

Article Title: Exploring the Epicarp Potential from Acrocomia aculeata Fruits: Chemical Analysis, Antioxidant and Antimicrobial Activities

doi: 10.3390/antiox14020181

Figure Lengend Snippet: Minimum inhibitory concentration and minimum bactericidal/fungicidal concentration of macaúba epicarp extracts/fractions and stilbenes (Sigma-Aldrich ® ).

Article Snippet: The MIC of the EtOAc fraction against Cryptococcus neoformans H99 serotype A (ATCC 208821) was determined in RPMI-MOPS medium according to the previously described broth microdilution method in the presence and absence of sorbitol solution (0.8 M) used as a fungal cell wall osmoprotectant.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Exploring the Epicarp Potential from Acrocomia aculeata Fruits: Chemical Analysis, Antioxidant and Antimicrobial Activities

doi: 10.3390/antiox14020181

Figure Lengend Snippet: Evaluation of the antifungal potential of stilbenes in Galleria mellonella larvae infected with C. neoformans H99. Larvae infected with 1 × 10 6 cells/larva were incubated in the dark, at room temperature, in sterilized Petri dishes for 1 h. They were then treated with 100 mg/kg of resveratrol or piceatannol.

Article Snippet: The MIC of the EtOAc fraction against Cryptococcus neoformans H99 serotype A (ATCC 208821) was determined in RPMI-MOPS medium according to the previously described broth microdilution method in the presence and absence of sorbitol solution (0.8 M) used as a fungal cell wall osmoprotectant.

Techniques: Infection, Incubation

Journal: Antioxidants

Article Title: Exploring the Epicarp Potential from Acrocomia aculeata Fruits: Chemical Analysis, Antioxidant and Antimicrobial Activities

doi: 10.3390/antiox14020181

Figure Lengend Snippet: Minimum inhibitory concentration and minimum bactericidal/fungicidal concentration of macaúba epicarp extracts/fractions and stilbenes (Sigma-Aldrich ® ).

Article Snippet: In addition, piceatannol exhibited significant fungicidal activity against all strains of Cryptococcus , while resveratrol exhibited inhibitory activity only against C. neoformans serotype D (ATCC 24067).

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Exploring the Epicarp Potential from Acrocomia aculeata Fruits: Chemical Analysis, Antioxidant and Antimicrobial Activities

doi: 10.3390/antiox14020181

Figure Lengend Snippet: Evaluation of the antifungal potential of stilbenes in Galleria mellonella larvae infected with C. neoformans H99. Larvae infected with 1 × 10 6 cells/larva were incubated in the dark, at room temperature, in sterilized Petri dishes for 1 h. They were then treated with 100 mg/kg of resveratrol or piceatannol.

Article Snippet: In addition, piceatannol exhibited significant fungicidal activity against all strains of Cryptococcus , while resveratrol exhibited inhibitory activity only against C. neoformans serotype D (ATCC 24067).

Techniques: Infection, Incubation

Journal: Antioxidants

Article Title: Exploring the Epicarp Potential from Acrocomia aculeata Fruits: Chemical Analysis, Antioxidant and Antimicrobial Activities

doi: 10.3390/antiox14020181

Figure Lengend Snippet: Minimum inhibitory concentration and minimum bactericidal/fungicidal concentration of macaúba epicarp extracts/fractions and stilbenes (Sigma-Aldrich ® ).

Article Snippet: The antibacterial and antifungal activities were evaluated against the Gram-negative bacteria Salmonella enterica serovar Typhimurium (ATCC 14028); Gram-positive bacteria Bacillus subtilis , Staphylococcus aureus (ATCC 6538), and methicillin-resistant Staphylococcus aureus (MRSA BMB 9393)—a clinical isolate obtained from Hospital Universitário Clementino Fraga Filho (HUCFF/UFRJ); the yeasts Candida albicans serotype B (ATCC 36802) and Cryptococcus neoformans serotype A (T 1 -444)—clinical isolate obtained from Universidade Federal de São Paulo, Brazil (UNIFESP—SP); Cryptococcus neoformans H99 serotype A (ATCC 208821)—provided by D.Sc.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Exploring the Epicarp Potential from Acrocomia aculeata Fruits: Chemical Analysis, Antioxidant and Antimicrobial Activities

doi: 10.3390/antiox14020181

Figure Lengend Snippet: Evaluation of the antifungal potential of stilbenes in Galleria mellonella larvae infected with C. neoformans H99. Larvae infected with 1 × 10 6 cells/larva were incubated in the dark, at room temperature, in sterilized Petri dishes for 1 h. They were then treated with 100 mg/kg of resveratrol or piceatannol.

Article Snippet: The antibacterial and antifungal activities were evaluated against the Gram-negative bacteria Salmonella enterica serovar Typhimurium (ATCC 14028); Gram-positive bacteria Bacillus subtilis , Staphylococcus aureus (ATCC 6538), and methicillin-resistant Staphylococcus aureus (MRSA BMB 9393)—a clinical isolate obtained from Hospital Universitário Clementino Fraga Filho (HUCFF/UFRJ); the yeasts Candida albicans serotype B (ATCC 36802) and Cryptococcus neoformans serotype A (T 1 -444)—clinical isolate obtained from Universidade Federal de São Paulo, Brazil (UNIFESP—SP); Cryptococcus neoformans H99 serotype A (ATCC 208821)—provided by D.Sc.

Techniques: Infection, Incubation