Studies have shown that the second N-terminal residue of fungal defensins is closely involved in the binding of defensins to lipid II, a bacterial cell wall precursor, and plays an important role in antibacterial activity. We found that the N-terminal residue is always aromatic in nature. In this study, 29 fungal defensin-like peptides were found via the genomic search strategy. Based on the type of aromatic residue at the second N-terminal site, we mainly divided these peptides into Phe, Trp, and Tyr types. We selected and characterized a defensin, blapersin, derived from Blastomyces percursus as a molecular model to investigate the functional significance of the N-terminal site. The native blapersin killed a wide spectrum of gram-positive bacteria at low molecular concentrations. Its aromatic mutants, W2F and W2Y, displayed enhanced antimicrobial activity, especially against the vancomycin-resistant Enterococcus faecium. The aromatic side chains containing Phe2 and Tyr2 seem to be more favorable for the antibacterial activity of blapersin those containing Trp2. However, the nonaromatic mutant W2A had almost no antibacterial activity. This indicates that the second N-terminal aromatic residue is essential for the antimicrobial action of blapersin. All these defensins have high stability and low toxicity. This is the first report on the enhancement of antibacterial activity by calibration of the N-terminal aromatic residue.

Sporothrix schenckii is a pathogenic dimorphic fungus with a global distribution. It grows in a multicellular hyphal form at 25˚C and a unicellular yeast form at 37˚C. The morphological switch from mold to yeast form is obligatory for establishing pathogenicity in S. schenckii. Two‑component signaling systems are utilized by eukaryotes to sense and respond to external environmental changes. DRK1is a hybrid histidine kinase, which functions as a global regulator of dimorphism and virulence in Blastomyces dermatitidis and Histoplasma capsulatum. An intracellular soluble hybrid histidine kinase, homologous to DRK1 in B. dermatitidis, has previously been identified in S. schenckii and designated as SsDRK1. In the present study, the function of SsDRK1 was investigated using double stranded RNA interference mediated by Agrobacterium tumefaciens. SsDRK1 was demonstrated to be required for normal asexual development, yeast‑phase cell formation, cell wall composition and integrity, melanin synthesis, transcription of the morphogenesis‑associated gene Ste20 that is involved in the high osmolarity glycerol/mitogen‑activated protein kinase pathway, and pathogenicity of S. schenckii in a murine model of cutaneous infection. Further investigations into the signals SsDRK1 responds to, and the interactions of upstream transmembrane hybrid histidine kinases with SsDRK1, are required to uncover novel targets for anti‑fungal therapies.

A novel dimorphic fungus, Emergomyces orientalis sp. nov. a close relative of systemic pathogens in the family Ajellomycetaceae (Blastomyces, Histoplasma). The fungus is reported in a 64-year-old male from Shanxi, China. The patient developed disseminated skin lesions, productive cough with fever and showed nodular opacities in his left lung on chest radiography. The patient had no identified cause of immunodeficiency apart from type-2 diabetes mellitus. Clinical, histopathological and mycological characteristics of the agent are given, and its phylogenetic position is determined with multilocus sequence data.

Recent discoveries of novel systemic fungal pathogens with thermally dimorphic yeast-like phases have challenged the current taxonomy of the Ajellomycetaceae, a family currently comprising the genera Blastomyces, Emmonsia, Emmonsiellopsis, Helicocarpus, Histoplasma, Lacazia and Paracoccidioides. Our morphological, phylogenetic and phylogenomic analyses demonstrated species relationships and their specific phenotypes, clarified generic boundaries and provided the first annotated genome assemblies to support the description of two new species. A new genus, Emergomyces, accommodates Emmonsia pasteuriana as type species, and the new species Emergomyces africanus, the aetiological agent of case series of disseminated infections in South Africa. Both species produce small yeast cells that bud at a narrow base at 37°C and lack adiaspores, classically associated with the genus Emmonsia. Another novel dimorphic pathogen, producing broad-based budding cells at 37°C and occurring outside North America, proved to belong to the genus Blastomyces, and is described as Blastomyces percursus.

In the present study, a new flavanoid 1, together with nine known ones 2-10 were isolated from the stem bark of Choerospondias axillaries, the fruit of which was used mainly for treatment of cardiovascular diseases in China. The structure of 1 was established on the basis of its extensive spectral data, and the absolute structures of 1 and 10 were determined by their CD data. The absolute structure of 10 was established for the first time. Among the obtained compounds, 5-8 inhibited the proliferation of K562 cells with inhibition rates of 26.6%, 65.7%, 40.4% and 45.6% at 100 µg/mL; 1 and 4-10 showed significant protective effects on anoxia-induced injury in cultured ECV304 or PC12 cells at 50 µg/mL; 8 and 9 showed antibacterial effects on Staphylococcus aureus ATCC6538 at the tested concentration of 150 µg/8 mm paper disc. Compounds 2 and 4-10 were isolated for the first time from this genus. The proliferation inhibiting activities of 7 and 8, the anti-hypoxia activities of 1 and 4-10, and the antibacterial effect of 8 and 9 on Staphylococcus aureus ATCC6538 are reported here for the first time.

The dimorphism of Sporothrix schenckii (S. schenckii) reflects a developmental switch in morphology and lifestyle that is necessary for virulence. DRK1, a hybrid histidine kinase, functions as a global regulator of dimorphism and virulence in Blastomyces dermatitidis (B. dermatitidis) and Histoplasma capsulatum (H. capsulatum). The partial cDNA sequence of DRK1 of S. schenckii, designated SsDRK1, was obtained using degenerate primers based on the conserved domain of the DRK1 of other fungi. The complete cDNA sequence of SsDRK1 was obtained by 5\' and 3\' RACE. The full-length cDNA is 4743 bp in size and has an open reading frame (ORF) of 4071 bp, encoding 1356 amino acid residues. The predicted molecular mass of SsDRK1 is 147.3 kDa with an estimated theoretical isoelectric point of 5.46. The deduced amino acid sequence of SsDRK1 shows 65% identity to that of B. dermatitidis. The SsDRK1 was predicted to be a soluble histidine kinase and to contain three parts: sensor domain, linker domain and functional domain. Quantitative real-time RT-PCR revealed that SsDRK1 was more highly expressed in the yeast stage compared with that in the mycelial stage, which indicated that the SsDRK1 may be involved in the dimorphic switch in S. schenckii.

OBJECTIVE: To detect peptide toxins in Amanita pallidorasea and to study the antifungal activities of peptide toxins against Blastomyces albicans.
METHODS: We separated and identified peptide toxins and determined its contents in the fruiting body, pileus and the mixture of stipe and volva from A. pallidorasea by HPLC and ESI-MS methods. Meanwhile, we detected antifungal activities of the crude toxin and the separated peptide toxins against Blastomyces albicans JLC31680 and JLC31681 by the paper disk method.
RESULTS: We totally got three peptide toxins: alpha-amanitin (alpha-AMA), beta-amanitin (beta-AMA) and phalloidin (PHD). The contents of alpha-AMA, beta-AMA and PHD were 30.3 mg/g, 6.99 mg/g and 9.95 mg/g in fruiting body, and 45.0 mg/g, 11.1 mg/g and 11.3 mg/g in pileus. The contents of alpha-AMA and PHD were 11.7 mg/g and 7.98 mg/g in the mixture of stipe and volva , but the beta-AMA was not detected in this part. The inhibition ratio of the crude toxin and alpha-AMA, beta-AMA and PHD to B. albicans JLC31680 were 11.96%, 32.52%, 23.29% (P<0.01) and 15.46% (P<0.05). The inhibition ratio of the crude toxin and beta-AMA to B. albicans JLC31681 was 10.16% and 11.10% (P < 0.01), while that of alpha-AMA\'s was 6.89% (P < 0.05).
CONCLUSIONS: A. pallidorasea is a new resource of peptide toxins with antifungal activity.