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1.
Molecules ; 26(21)2021 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-34770899

RESUMO

The extracellular secreted protein of Bifidobacterium longum (B. longum) plays an important role in maintaining the homeostasis of the human intestinal microenvironment. However, the mechanism(s) of interaction remain unclear. Lysozyme is a kind of antibacterial peptide. In this study, the amino acid sequence of a lysozyme-like protein of B. longum based on whole-genome data of an isolate from human gut feces was found. We further predicted functional domains from the amino acid sequence, purified the protein, and verified its bioactivity. The growth of some bacteria were significantly delayed by the 020402_LYZ M1 protein. In addition, the gut microbiota was analyzed via high-throughput sequencing of 16S rRNA genes and an in vitro fermentation model, and the fluctuations in the gut microbiota under the treatment of 020402_LYZ M1 protein were characterized. The 020402_LYZ M1 protein affected the composition of human gut microbiota significantly, implying that the protein is able to communicate with intestinal microbes as a regulatory factor.


Assuntos
Proteínas de Bactérias/metabolismo , Bifidobacterium/metabolismo , Microbioma Gastrointestinal , Interações Microbianas , Proteínas de Bactérias/química , Bifidobacterium/enzimologia , Biologia Computacional/métodos , Fezes/microbiologia , Humanos , Modelos Moleculares , Proteoma , Proteômica/métodos , Relação Estrutura-Atividade
2.
Theranostics ; 14(4): 1764-1780, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38389846

RESUMO

Rationale: The present understanding of the cellular characteristics and communications in crystal nephropathy is limited. Here, molecular and cellular studies combined with single-cell RNA sequencing (scRNA-seq) were performed to investigate the changes in cell components and their interactions in glyoxylate-induced crystallized kidneys to provide promising treatments for crystal nephropathy. Methods: The transcriptomes of single cells from mouse kidneys treated with glyoxylate for 0, 1, 4, or 7 days were analyzed via 10× Genomics, and the single cells were clustered and characterized by the Seurat pipeline. The potential cellular interactions between specific cell types were explored by CellChat. Molecular and cellular findings related to macrophage-to-epithelium crosstalk were validated in sodium oxalate (NaOx)-induced renal tubular epithelial cell injury in vitro and in glyoxylate-induced crystal nephropathy in vivo. Results: Our established scRNA atlas of glyoxylate-induced crystalline nephropathy contained 15 cell populations with more than 40000 single cells, including relatively stable tubular cells of different segments, proliferating and injured proximal tubular cells, T cells, B cells, and myeloid and mesenchymal cells. In this study, we found that Mrc1+ macrophages, as a subtype of myeloid cells, increased in both the number and percentage within the myeloid population as crystal-induced injury progresses, and distinctly express IGF1, which induces the activation of a signal pathway to dominate a significant information flow towards injured and proliferating tubule cells. IGF1 promoted the repair of damaged tubular epithelial cells induced by NaOx in vitro, as well as the repair of damaged tubular epithelial cells and the recovery of disease outcomes in glyoxylate-induced nephrolithic mice in vivo. Conclusion: After constructing a cellular atlas of glyoxylate-induced crystal nephropathy, we found that IGF1 derived from Mrc1+ macrophages attenuated crystal nephropathy through promoting renal tubule cell proliferation via the AKT/Rb signaling pathway. These findings could lead to the identification of potential therapeutic targets for the treatment of crystal nephropathy.


Assuntos
Nefropatias , Proteínas Proto-Oncogênicas c-akt , Animais , Camundongos , Proliferação de Células , Glioxilatos , Nefropatias/metabolismo , Macrófagos/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais
3.
Microorganisms ; 11(6)2023 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-37374932

RESUMO

Sorghum bicolor is cultivated worldwide. Leaf spot of sorghum, which leads to leaf lesions and yield reduction, is a prevalent and serious disease in Guizhou Province, southwest China. In August 2021, new leaf spot symptoms were observed on sorghum leaves. In this study, traditional methods and modern molecular biology techniques were used to isolate and identify the pathogen. Sorghum inoculated with the isolate GY1021 resulted in reddish brown lesion that similar to symptoms observed in the field: the original isolate inoculated was reisolated and Koch's postulates were fulfilled. Based on morphological features and phylogenetic analysis of the internal transcribed spacer (ITS) combined sequence with ß-tubulin (TUB2) and translation elongation factor 1-α (TEF-1α) genes, the isolate was identified as Fusarium thapsinum (Strain accession: GY 1021; GenBank Accession: ITS (ON882046), TEF-1α (OP096445), and ß-TUB (OP096446)). Then, we studied the bioactivity of various natural products and microorganisms against F. thapsinum using the dual culture experiment. Carvacrol, 2-allylphenol, honokiol, and cinnamaldehyde showed excellent antifungal activity, with EC50 values of 24.19, 7.18, 46.18, and 52.81 µg/mL, respectively. The bioactivity of six antagonistic bacteria was measured using a dual culture experiment and the mycelial growth rate method. Paenibacillus polymyxa, Bacillus amyloliquefaciens and Bacillus velezensis displayed significant antifungal effects against F. thapsinum. This study provides a theoretical basis for the green control of leaf spot of sorghum.

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