Project description:Cryptococcus neoformans is a fungal pathogen responsible for an increased mortality among immunocompromised individuals. Long antifungal therapies to treat cryptococcal infections have compounded the occurrence of resistant strains that threaten the efficacy of current treatments. In this senseDue to resistance mechanisms, discovery of compounds that inhibit virulence factors, rather than kill the fungus, have emerged as potential new strategies to combat infection and reduce the rate of resistance due to lower selective pressure. Invertebrates rely solely on an effective innate immune system to prevent infections, provide providing a potential one health approach for discovery of novel antifungal and antibacterial compounds. Here, we demonstrate a differentiated extraction of proteins from three mollusks (freshwater and terrestrial) and evaluate extract their effects against the growth and virulence factor production (thermotolerance, melanin, capsule, and biofilm) in C. neoformans. We show that clarified extracts of Planorbella pilsbryi have a fungicidal effect on cryptococcal cells in a comparable way to Fluconazolefluconazole. Similarly, crude and clarifiedall extracts of Cipangopaludina chinensis not only affects cryptococcal thermotolerance but also impairs biofilm and capsule production. In addition, incubation of C. neoformans with clarified extracts of Cepaea nemoralis extracts reduced capsule production. Using inhibitory activity of extracts against peptidases related with virulence factors and Quantitative quantitative proteomics arose distinct proteome signatures for each extract and proposed proteins driving the observed anti-virulence properties. Overall, this work demonstratesproves the potential of compounds derived from natural sources to inhibit virulence factor production in a clinically important fungal pathogen.
Project description:The study investigates the effects of myrobalane fruit extracts, essential in Asian traditional medicine and notably part of the Triphala formulation, on human cell models. The complexity of these botanical preparations suggests a multi-target mode of action, making it difficult to identify specific active ingredients. The in vitro study revealed that, beyond their antioxidant properties, myrobalane fruit extracts modulate tryptophan metabolism and affect immunobiochemical and cytoprotective signaling pathways in a dose-dependent manner. Integrated transcriptome analysis of treated cells showed impacts on immune response pathways, oxidative stress, and detoxification processes. Specifically, a synergistic activation of the endogenous antioxidant response was observed in liver epithelial cells treated with a combination of the three fruit extracts. These findings highlight the modulation of various signaling pathways and cellular processes, underpinning the complex multi-target effects of myrobalane fruit extracts. Despite being limited to in vitro data, this study enhances the understanding of the mode of action of these botanical mixtures.
Project description:We investigate the molecular targets and pathways that the neem extracts and the associated compounds act through, to bring about tumor suppression by using a genome-wide functional pooled shRNA screen on head and neck squamous cell carcinoma cell line treated with crude neem leaf extracts. We analyzed differences in global clonal sizes of the shRNA-infected cells cultured under no treatment and treatment with neem leaf extract conditions, assayed using next-generation sequencing. We further analyzed differences in gene expression in HSC-4 cells, upon treatment with neem leaf extract in a time-dependent manner, followed by a time-dependent rescue. Our results indicate that neem extract simultaneously affects various important molecular signaling pathways in head and neck cancer cells, some (TGF-β/HSF-1) of which may be therapeutic targets for this devastating tumor.
Project description:In this study, we have characterized and compared the effects of differently prepared chamomile extracts and characteristic pure compounds on the T cell redox milieu as well as on the migration, activation, proliferation, and cytokine production of primary human T cells. Futhermore, nCounter based gene expression profiling was performed on the most promising extracts (Chamomile aqueous total fermented-CT, aqueous toot fermented-CR, and ethanolic flower-CF) and pure compounds (Apigenin-Ap and Chamazulene-Cz) to identify the genes related to different T cell functions, that were targeted by the different treatment conditions.
Project description:To understand how GIPC3 exerts its effects on cuticular plate dimensions, we examined the GIPC3 protein-interaction network in hair cells. We immunoaffinity purified GIPC3 from crosslinked chicken inner ear extracts that were enriched for stereocilia, but still contain large amounts of hair-cell cytoplasmic proteins (Morgan et al., 2016). We carried out two separate experiments, each with ~1000 chicken ears, where we stabilized protein complexes using primary amine-reactive homo-bifunctional N-hydroxysuccimide ester crosslinkers that are thiol-cleavable and hence reversible (Mattson et al., 1993). In one experiment, we used dithiobis(succinimidyl propionate) (DSP), a membrane-permeable crosslinker that crosslinks extracellular and intracellular complexes; in the other experiment, we used 3,3'-dithiobis(sulfosuccinimidyl propionate) (DTSSP), which is membrane impermeant and thus only stabilizes extracellular and transmembrane complexes. We prepared soluble extracts of crude, crosslinked stereocilia and used these fractions for identifying GIPC3-interacting proteins.