Extracts from submerged cultured mycelium of two strains of Flammulina velutipes, a popular culinary mushroom, were obtained by ultrasound and tested in vitro to determine their activity in innate immunity (monocytes/ macrophages). In addition, polyclonal antibodies against the extracts were produced. Both extracts have similar glycoproteins that contain mannose and glucose but have different glycoproteins with galactoseamine units. Two novel immunogenic glycoproteins with molecular weights of 32 and 25 kDa have been revealed. It is thought that these proteins are produced only by submerged cultured mycelium. Both extracts show immune-enhancing activity based on the significant modification of various parameters such as cytokine production, phagocytosis, and reactive oxygen species production.
The development of mature biofilms is an aid in numerous aspects of the life cycle of fungi. It is well known that Sporothrix schenckii complex causes a benign subcutaneous mycosis, but recent studies have suggestedthat biofilm formation may be one of the important factors involved in its virulence. Here we report the study of the biomass organization and a model of the stages of S. schenckii biofilm development: adsorption, active adhesion, microcolony formation, maturation, and dispersal of biofilm fragments. During the development, the biofilm is surrounded by extracellular matrix, which contains glycoprotein (mannose rich), carbohydrates, lipids, and nucleic acid. In addition, the extracellular DNA increases in extracellular matrix as a key component to structural integrity and antifungal resistance. The study of S. schenckii biofilm contributes to a better understanding of growth biofilm and physiology, adding new insights into the mechanisms of virulence and persistence of pathogenic microorganisms.
Background:Recently, extracts from natural sources have been tested for their antifungal properties. In this aspect, Flammulina velutipes extracts possess a significant amount of branch-chained carbohydrates with mannose moieties that, hypothetically, can reduce the adhesion.Objective:In this study, we assessed the capacity of extracts from F. velutipes (wild-type AQF-1 and ATCC 34574 as the reference strain) to inhibit the adhesion of S. schenkii and C. albicans to epithelial cells.Materials and Methods:The aqueous extracts from F. velutipes strains were obtained by sonication, total carbohydrate and protein was analyzed by Dubois and Lowry methods respectively. Effect of the extracts (50, 100 and 150 μg/mL) on the fungi adhesion to host cells was evaluated after 1 h interaction, and the percentage of inhibition of adhesion was measured. After of interaction the cytoskeleton from cell was analyzed with phalloidin-FITC.Results:The extract from strain AQF-1 (50, 100 and 150 μg/mL) inhibited the adhesion of: S. schenkii in a dose-dependent manner (4.9, 7.5 and 12.7%, respectively) and C. albicans in a dose-independent manner (5.2%). The percentage of inhibition by extracts from the strain ATCC34574 at the same concentrations, shown that are dose independent for both fungi: 3.9% for S. schenkii and 2.6% for C. albicans.Conclusion:The extracts from F. velutipes inhibit the adhesion of pathogenic fungi to host cells. The mechanism molecular is unknown; however, is probably an interaction between the polysaccharides from extracts with the fungi receptors. This aspect is currently analyzed.SUMMARY
The yields of mycelium from two strains of F. velutipes and the extract from it were similar.Extracts from both strains have inhibited adhesion of S. schenkii and C. albicans to epithelial cells in vitro, but the extract from strain AQF-1 was more effective.The extracts have not prevented damage to epithelial cells caused by pathogenic fungi.
Abbreviation Used: YPG: Yeast peptone glucose, DMEM: Dulbecco's Modified Eagle's medium, FITC: Fluorescein isothiocyanate
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