Identification of a Surface Protein from Lactobacillus reuteri JCM1081 That Adheres to Porcine Gastric Mucin and Human Enterocyte-Like HT-29 Cells

Adhesion of lactobacilli to the host gastrointestinal (GI) tract is considered an important factor in health-promoting effects. However, studies addressing the molecular mechanisms of the adhesion of lactobacilli to the host GI tract have not yet been performed. The aim of this work was to identify Lactobacillus reuteri surface molecules mediating adhesion to intestinal epithelial cells and mucins. Nine strains of lactobacilli were tested for their ability to adhere to human enterocyte-like HT-29 cells. The cell surface proteins involved in the adhesion of Lactobacillus to HT-29 cells and gastric mucin were extracted. The active fractions were detected by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and Western blotting with horseradish peroxidase-labeled mucin and NHS-Biotin-labeled HT-29 cells. Furthermore, tandem mass spectrometry analysis was performed to identify the surface protein that participates in adhesion. It was shown that the ability of lactobacilli to adhere to HT-29 cells in vitro varied considerably among different strains. The most adhesive strain was the chicken intestinal tract isolate Lactobacillus reuteri JCM1081 (495.07 ± 80.03 bacterial cells/100 HT-29 cells). The adhesion of L. reuteri JCM1081 to HT-29 cells appeared to be mediated by a cell surface protein, with an approximate molecular mass of 29 kDa. The peptides generated from the 29-kDa protein significantly matched the Lr0793 protein sequence of L. reuteri strain ATCC55730 (∼71.1% identity) and displayed significant sequence similarity to the putative ATP-binding cassette transporter protein CnBP

Chemical composition of the essential oil of whole plant of Elsholtizia dense Benth and its anti-tumor effect on human hepatoma cells

Purpose: To determine the chemical components of the essential oil of Elsholtizia dense in Sichuan Province and evaluate the effect of the oil on human hepatoma cells (SMMC-7721) in vitro.Methods: The essential oil was extracted using the modified steam-distillation  extraction method, and its chemical components were determined by gas  chromatography-mass spectrometry (GC-MS). The effect of the essential oil on proliferation of SMMC-7721 cells was studied by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, with L02 and HeLa cells serving as control groups.Results: GC-MS results show that the essential oil of E. dense contains 40  components. Thirty seven components were identified and accounted for 98.39 % of the essential oil. The two main components were rosefuran epoxide (53.12 %) and 2-ethyl imidazole (29.8 %). The oil significantly inhibited cell proliferation in a concentration- and time-dependent manner (p < 0.05). SMMC-7721 cells were more inhibited than L02 and HeLa cells by the oil, with half maximal inhibitory concentration (IC50) values of 26.23 and 25.46 μg/mL after 8-h and 24-h treatments, respectively.Conclusion: Out of the 40 chemical components of the essential oil of E. dense, rosefuran epoxide and 2-ethyl imidazole were the most abundant. The oil has a significant anti-tumor effect on SMMC-7721 cells, and thus has a potential to be developed as an anti-liver cancer drug.Keywords: Medicinal herb, Elsholtizia dense Benth, Essential oil, Rosefuran epoxide, 2-Ethyl imidazole, Anti-tumor activit

In Vivo Disruption of TGF-β Signaling by Smad7 in Airway Epithelium Alleviates Allergic Asthma but Aggravates Lung Carcinogenesis in Mouse

BACKGROUND: TGF-beta has been postulated to play an important role in the maintenance of epithelial homeostasis and the development of epithelium-derived cancers. However, most of previous studies are mainly focused on the function of TGF-beta in immune cells to the development of allergic asthma and how TGF-beta signaling in airway epithelium itself in allergic inflammation is largely unknown. Furthermore, the in vivo TGF-beta function specifically in the airway epithelium during lung cancer development has been largely elusive. METHODOLOGY/PRINCIPAL FINDINGS: To evaluate the in vivo contribution of TGF-beta signaling in lung epithelium to the development of allergic disease and lung cancer, we generated a transgenic mouse model with Smad7, an intracellular inhibitor of TGF-beta signaling, constitutively expressed in mouse airway Clara cells using a mouse CC10 promoter. The mice were subjected to the development of OVA-induced allergic asthma and urethane-induced lung cancer. The Smad7 transgenic animals significantly protected from OVA-induced asthma, with reduced airway inflammation, airway mucus production, extracellular matrix deposition, and production of OVA-specific IgE. Further analysis of cytokine profiles in lung homogenates revealed that the Th2 cytokines including IL-4, IL-5 and IL-13, as well as other cytokines including IL-17, IL-1, IL-6, IP10, G-CSF, and GM-CSF were significantly reduced in the transgenic mice upon OVA induction. In contrast, the Smad7 transgenic animals had an increased incidence of lung carcinogenesis when subjected to urethane treatment. CONCLUSION/SIGNIFICANCE: These studies, therefore, demonstrate for the first time the in vivo function of TGF-beta signaling specifically in airway epithelium during the development of allergic asthma and lung cancer

3,6-Didehydro-5-hy­droxy-1,2-O-iso­propyl­idene-5-C-nitro­meth­yl-α-d-gluco­furan­ose

The title compound, C10H15NO7, consists of one methyl­enedi­oxy ring and two fused tetra­hydro­furan rings. The three fused rings exhibit cis arrangements at the ring junctions. One O atom of a tetra­hydro­furan ring and the H atoms bound to the neighboring C atoms are disordered over two orientations with site-occupancy factors of 0.69 (1) and 0.31 (1). intra­molecular O—H⋯O and C—H⋯O inter­actions stabilize the mol­ecular conformation. In the crystal structure, inter­molecular O—H⋯O and C—H⋯O inter­actions link the mol­ecules into a three-dimensional network

Zanthoxylum bungeanum seed oil inhibits tumorigenesis of human melanoma A375 by regulating CDC25A/CyclinB1/CDK1 signaling pathways in vitro and in vivo

Background:Zanthoxylum bungeanum seed oil (ZBSO) is extracted from the seeds of the traditional Chinese medicine Z. bungeanum Maxim, which has been shown to have anti-melanoma effects. However, the specific mechanisms are not illustrated adequately.Aims: To further investigate the mechanism by which ZBSO inhibits melanoma and to provide scientific evidence to support ZBSO as a potential melanoma therapeutic candidate.Methods: CCK-8 assays were used to detect the function of ZBSO on A375 cells. Based on transcriptomics analyses, Western blot analysis was applied to determine whether an association existed in ZBSO with the CDC25A/CyclinB1/CDK1 signaling pathway. In addition, RT-qPCR and immunohistochemistry analysis validated that ZBSO has the anti-melanoma effect in a nude mouse xenograft model of human melanoma. Then, 16S rRNA sequencing was used to detect the regulation of gut microbes.Results: Cellular assays revealed that ZBSO could inhibit A375 cell viability by regulating the cell cycle pathway. Further studies presented that ZBSO could constrain CDC25A/CyclinB1/CDK1 signaling pathway in vitro and in vivo models of melanoma. ZBSO did not produce toxicity in mice, and significantly reduced tumor volume in xenotransplants of A375 cells. Genome analysis indicated that ZBSO successfully altered specific gut microbes.Conclusion: ZBSO inhibited the growth of A375 cells by regulating CDC25A/cyclinB1/CDK1 signaling pathway both in vitro and in vivo, suggesting that ZBSO may be a novel potential therapeutic agent

Crosstalk between microbial biofilms in the gastrointestinal tract and chronic mucosa diseases

The gastrointestinal (GI) tract is the largest reservoir of microbiota in the human body; however, it is still challenging to estimate the distribution and life patterns of microbes. Biofilm, as the predominant form in the microbial ecosystem, serves ideally to connect intestinal flora, molecules, and host mucosa cells. It gives bacteria the capacity to inhabit ecological niches, communicate with host cells, and withstand environmental stresses. This study intends to evaluate the connection between GI tract biofilms and chronic mucosa diseases such as chronic gastritis, inflammatory bowel disease, and colorectal cancer. In each disease, we summarize the representative biofilm makers including Helicobacter pylori, adherent-invasive Escherichia coli, Bacteroides fragilis, and Fusobacterium nucleatum. We address biofilm’s role in causing inflammation and the pro-carcinogenic stage in addition to discussing the typical resistance, persistence, and recurrence mechanisms seen in vitro. Biofilms may serve as a new biomarker for endoscopic and pathologic detection of gastrointestinal disease and suppression, which may be a useful addition to the present therapy strategy

Component Interaction of ESCRT Complexes Is Essential for Endocytosis-Dependent Growth, Reproduction, DON Production and Full Virulence in Fusarium graminearum

Multivesicular bodies (MVBs) are critical intermediates in the trafficking of ubiquitinated endocytosed surface proteins to the lysosome/vacuole for destruction. Recognizing and packaging ubiquitin modified cargoes to the MVB pathway require ESCRT (Endosomal sorting complexes required for transport) machinery, which consists of four core subcomplexes, ESCRT-0, ESCRT-I, ESCRT-II, and ESCRT-III. Fusarium graminearum is an important plant pathogen that causes head blight of major cereal crops. Our previous results showed that ESCRT-0 is essential for fungal development and pathogenicity in Fusarium graminearum. We then, in this study, systemically studied the protein-protein interactions within F. graminearum ESCRT-I, -II or -III complex, as well as between ESCRT-0 and ESCRT-I, ESCRT-I and ESCRT-II, and ESCRT-II and ESCRT-III complexes and found that loss of any ESCRT component resulted in abnormal function in endocytosis. In addition, ESCRT deletion mutants displayed severe defects in growth, deoxynivalenol (DON) production, virulence, sexual, and asexual reproduction. Importantly genetic complementation with corresponding ESCRT genes fully rescued all these defective phenotypes, indicating the essential role of ESCRT machinery in fungal development and plant infection in F. graminearum. Taken together, the protein-protein interactome and biological functions of the ESCRT machinery is first profoundly characterized in F. graminearum, providing a foundation for further exploration of ESCRT machinery in filamentous fungi

Fish Oil Enhances Recovery of Intestinal Microbiota and Epithelial Integrity in Chronic Rejection of Intestinal Transplant

The intestinal chronic rejection (CR) is the major limitation to long-term survival of transplanted organs. This study aimed to investigate the interaction between intestinal microbiota and epithelial integrity in chronic rejection of intestinal transplantation, and to find out whether fish oil enhances recovery of intestinal microbiota and epithelial integrity.. In addition, CR rats showed pronounced alteration of tight junction, depicted by marked changes in epithelial cell ultrastructure and redistribution of occuldin and claudins as well as disruption in TJ barrier function. Fish oil administration ameliorated disruption of epithelial integrity in CR, which was associated with an improvement of the mucosal structure leading to improved tight junctions.Our study have presented novel evidence that fish oil is involved in the maintenance of epithelial TJ integrity and recovery of gut microbiota, which may have therapeutic potential against CR in intestinal transplantation