Project description:Multi-drug resistant bacteria are not effectively managed with current treatments, making it a serious global problem. Therefore, there is an essential need for finding new antimicrobial agents. In this regard, silver nanoparticles (Ag-NPs) have been projected as a new generation of antimicrobial agents. Ag-NPs were biomediated by Egyptian Streptomyces roseolus for the first time, which was molecularly identified using 16S rRNA sequencing under accession no. MT071505. Biosynthesized Ag-NPs were characterized using UV-Vis spectroscopy, XRD, TEM, FTIR, and DLS. FTIR analysis confirmed the presence of different bioactive functional groups, such as N-H, C-H, C-O-C, C-NH2, and C=O acting as bioreducing/stabilizing agents for Ag-NPs. Ag-NPs exhibited antimicrobial activity against some multi-drug resistant Gram-positive and Gram-negative pathogens. MBC of Ag-NPs against Listeria monocytogenes and Klebsiella pneumonia were 0.195 and 0.048 mg/mL, respectively, with a tolerance level of 2 confirming its biocidal effect. SEM imaging of Ag-NPs-treated L. monocytogenes and K. pneumonia showed shrunk destroyed cells after 6 h of treatment. Biosynthesized Ag-NPs exhibited IC50 of < 0.3 and 8.21 mg/mL, on normal Human Skin Fibroblast, and Blood Lymphocytes, respectively. IC50 values were significantly higher than its MBC values, with no harmful cytotoxic effect, thus can be safely applied at its biocidal concentration. For biosafety purposes, the genotoxicity of biosynthesized Ag-NPs was assessed using Comet assay for the first time on Blood Lymphocytes, with zero-tail and 100% head intensity indicating non-genotoxic effect. An ecofriendly biomediated synthesis of Ag-NPs was described with easy scale-up, non-toxic by-products, so, it can be recommended as a powerful-safe antimicrobial agent.
Project description:Frequently observed in tropical and sub-tropical regions, crops contamination by aflatoxin B1 (AFB1) produced by Aspergillus flavus, is emerging in Europe, due to climate change. Many alternative methods are currently developed to reduce the use of chemical inputs to prevent mycotoxin contamination, such as biocontrol agents (BCAs). Actinobacteria are known to produce many bioactive compounds and some of them are able to reduce in vitro AFB1 concentration. In this context, the present study aims to analyze the effect of a cell free supernatant (CFS) from Streptomyces roseolus liquid culture on A. flavus development, as well as on its transcriptome profile using microarray assay and its impact on AFB1 concentration. To study the impact of Streptomyces roseolus cell free supernatant on global transcriptome of Aspergillus flavus we have employed whole genome microarray expression profiling.
Project description:Crop contamination by aflatoxin B1 (AFB1), an Aspergillus-flavus-produced toxin, is frequently observed in tropical and subtropical regions. This phenomenon is emerging in Europe, most likely as a result of climate change. Alternative methods, such as biocontrol agents (BCAs), are currently being developed to reduce the use of chemicals in the prevention of mycotoxin contamination. Actinobacteria are known to produce many bioactive compounds, and some of them can reduce in vitro AFB1 concentration. In this context, the present study aims to analyze the effect of a cell-free supernatant (CFS) from Streptomyces roseolus culture on the development of A. flavus, as well as on its transcriptome profile using microarray assay and its impact on AFB1 concentration. Results demonstrated that in vitro, the S. roseolus CFS reduced the dry weight and conidiation of A. flavus from 77% and 43%, respectively, and was therefore associated with a reduction in AFB1 concentration reduction to levels under the limit of quantification. The transcriptomic data analysis revealed that 5198 genes were differentially expressed in response to the CFS exposure and among them 5169 were downregulated including most of the genes involved in biosynthetic gene clusters. The aflatoxins' gene cluster was the most downregulated. Other gene clusters, such as the aspergillic acid, aspirochlorine, and ustiloxin B gene clusters, were also downregulated and associated with a variation in their concentration, confirmed by LC-HRMS.
Project description:HnRNPA2B1 encodes an RNA binding protein associated with neurodegenerative disorders. However, its function in the nervous system is unclear. Transcriptome-wide cross-linking and immunoprecipitation in mouse spinal cord discover UAGG motifs enriched within ~2,500 hnRNP A2/B1 binding sites and an unexpected role for hnRNP A2/B1 in alternative polyadenylation. Loss of hnRNP A2/B1 results in alternative splicing, including skipping of an exon in amyotrophic lateral sclerosis (ALS)-associated D-amino acid oxidase (DAO) that reduces D-serine metabolism. Inclusion of the DAO exon is also reduced in transgenic ALS mice models. ALS-associated hnRNP A2/B1 D290V mutant patient fibroblasts and motor neurons differentiated from induced pluripotent stem cells demonstrate gain-of-mutant-dependent splicing differences. Mutant motor neurons also exhibit increased hnRNP A2/B1 localization to cytoplasmic granules during stress, which are abrogated by a small molecule CA43. Our findings and cellular resource identify RNA networks affected in loss of normal and mutated hnRNP A2/B1 with broad relevance to neurodegeneration.
Project description:Extracts of Streptomyces sp. P9-A2 from Jaegersborg Dyrehaven, Denmark. Agar plug extracts from the inhibition zone of P9-A2 and Aspergillus tubingensis.