Journal: Frontiers in cellular and infection microbiology

Article Title: Production of Membrane Vesicles by Enterococcus faecium Cultured With or Without Subinhibitory Concentrations of Antibiotics and Their Pathological Effects on Epithelial Cells.

doi: 10.3389/fcimb.2019.00295

Figure Lengend Snippet: FIGURE 2 | Proteomic analysis of MVs produced by E. faecium ATCC 700221. MVs were isolated from E. faecium cultured in BHI broth (MVs/BHI), BHI broth with 256 µg/ml vancomycin (MVs/VAN), or BHI broth with 1 µg/ml linezolid (MVs/LIN). (A,B) A total of 438 proteins in the MVs/BHI were analyzed based on cellular localization (A) and Gene Ontology (B). (C) A Venn diagram of proteins identified in the MVs/BHI, MVs/VAN, and MVs/LIN.

Article Snippet: MVs produced by E. faecium ATCC 700221 were isolated from bacterial culture supernatants as previously described (Gurung et al., 2011; Kim et al., 2016; Yun et al., 2018).

Techniques: Produced, Isolation, Cell Culture

Journal: Frontiers in cellular and infection microbiology

Article Title: Production of Membrane Vesicles by Enterococcus faecium Cultured With or Without Subinhibitory Concentrations of Antibiotics and Their Pathological Effects on Epithelial Cells.

doi: 10.3389/fcimb.2019.00295

Figure Lengend Snippet: FIGURE 1 | Membrane vesicles (MVs) produced by E. faecium ATCC 700221 cultured with or without antibiotics. (A) Transmission electron micrographs of MVs from E. faecium cultured in BHI broth. (B) SDS-PAGE analysis of proteins obtained from bacterial lysates (lane 1), culture supernatant (lane 2), and MVs (lane 3). Bacteria were cultured in BHI broth to late exponential phase. Lane M, molecular weight marker. (C) Production of MVs from E. faecium cultured with or without antibiotics. MVs were isolated from E. faecium cultured in BHI broth (MVs/BHI), BHI broth with 256 µg/ml vancomycin (MVs/VAN), or BHI broth with 1 µg/ml linezolid (MVs/LIN). The protein concentration of MVs isolated from 1 L of bacterial culture was measured using a modified BCA assay. Data are presented as the mean ± SD of three independent experiments. **P < 0.01 compared to MVs/BHI. (D) SDS-PAGE analysis of MV proteins. Lane M, molecular weight marker; 1, MVs/BHI; 2, MVs/VAN; 3, MVs/LIN. (A,B,D) represent one of three independent experiments.

Article Snippet: MVs produced by E. faecium ATCC 700221 were isolated from bacterial culture supernatants as previously described (Gurung et al., 2011; Kim et al., 2016; Yun et al., 2018).

Techniques: Membrane, Produced, Cell Culture, Transmission Assay, SDS Page, Bacteria, Molecular Weight, Marker, Isolation, Protein Concentration, BIA-KA

Journal: Frontiers in cellular and infection microbiology

Article Title: Production of Membrane Vesicles by Enterococcus faecium Cultured With or Without Subinhibitory Concentrations of Antibiotics and Their Pathological Effects on Epithelial Cells.

doi: 10.3389/fcimb.2019.00295

Figure Lengend Snippet: FIGURE 3 | Cytotoxicity of Caco-2 cells treated with MVs from E. faecium ATCC 700221. MVs were isolated from culture supernatants of E. faecium cultured in BHI broth (MVs/BHI), BHI broth with 256 µg/ml vancomycin (MVs/VAN), or BHI broth with 1 µg/ml linezolid (MVs/LIN). Cells were treated with various concentrations of E. faecium MVs for 24 h. (A) Cell viability was determined using an MTT assay. Data are presented as the mean ± SD of three independent experiments. +P < 0.05, ++P < 0.01 compared to untreated control cells. *P < 0.05, **P < 0.01 among the same concentrations of MVs/BHI, MVs/VAN, or MVs/LIN. (B) Flow cytometric analysis of Caco-2 cell death induced by E. faecium MVs. Cells were treated with 20 µg/ml of E. faecium MVs for 24 h. Cells were stained with FITC-Annexin V and propidium iodide, and 104 cells were counted. The figure represents one of three independent experiments that yielded similar results.

Article Snippet: MVs produced by E. faecium ATCC 700221 were isolated from bacterial culture supernatants as previously described (Gurung et al., 2011; Kim et al., 2016; Yun et al., 2018).

Techniques: Isolation, Cell Culture, MTT Assay, Control, Staining

Journal: Frontiers in cellular and infection microbiology

Article Title: Production of Membrane Vesicles by Enterococcus faecium Cultured With or Without Subinhibitory Concentrations of Antibiotics and Their Pathological Effects on Epithelial Cells.

doi: 10.3389/fcimb.2019.00295

Figure Lengend Snippet: FIGURE 5 | Host cell responses to proteinase K (PK)-treated E. faecium MVs. (A) Caco-2 cells were incubated with intact or PK-treated MVs/BHI for 24 h and cell viability was determined using an MTT assay. Data are presented as the mean ± SD of three independent experiments. (B) Cells were incubated with intact or PK-treated MVs/BHI for 3 h and gene expression was assessed via qPCR. Data are presented as the mean ± SD of three independent experiments. **P < 0.01 comparing intact and PK-treated MVs.

Article Snippet: MVs produced by E. faecium ATCC 700221 were isolated from bacterial culture supernatants as previously described (Gurung et al., 2011; Kim et al., 2016; Yun et al., 2018).

Techniques: Incubation, MTT Assay, Gene Expression

Journal: Frontiers in cellular and infection microbiology

Article Title: Production of Membrane Vesicles by Enterococcus faecium Cultured With or Without Subinhibitory Concentrations of Antibiotics and Their Pathological Effects on Epithelial Cells.

doi: 10.3389/fcimb.2019.00295

Figure Lengend Snippet: FIGURE 4 | Expression of pro-inflammatory cytokine and chemokine genes in Caco-2 cells treated with MVs from E. faecium ATCC 700221. MVs were isolated from culture supernatants of E. faecium cultured in BHI broth (MVs/BHI), BHI broth with 256 µg/ml vancomycin (MVs/VAN), or BHI broth with 1 µg/ml linezolid (MVs/LIN). Cells were treated with various concentrations of E. faecium MVs for 3 h and gene expression was assessed via qPCR. Data are presented as the mean ± SD of three independent experiments. +P < 0.05, ++P < 0.01 compared to untreated control cells. *P < 0.05, **P< 0.01 among the same concentrations of MVs/BHI, MVs/VAN, or MVs/LIN.

Article Snippet: MVs produced by E. faecium ATCC 700221 were isolated from bacterial culture supernatants as previously described (Gurung et al., 2011; Kim et al., 2016; Yun et al., 2018).

Techniques: Expressing, Isolation, Cell Culture, Gene Expression, Control

Journal: bioRxiv

Article Title: The first digital twin of Enterococcus faecium metabolism reproduces high-throughput phenotyping data

doi: 10.64898/2026.05.01.720924

Figure Lengend Snippet: Schematic representation of a section of the fatty acid, phospholipid and lipoteichoic acid (LTA) biosynthesis pathways in E. faecium DO, as visualized by Escher. Enzymes are shown in blue, while key metabolites are highlighted in red. The pathways include the conversion of acyl-ACP to phosphatidic acid, CDP-diacylglycerol, phosphatidylglycerol, cardiolipin, and the synthesis of LTA from diacylglycerol.

Article Snippet: The strain whose genome was used in this reconstruction is E. faecium TX0016, also known as the DO strain (ATCC BAA-472), isolated from the blood of a patient with infective endocarditis.

Techniques:

Journal: bioRxiv

Article Title: The first digital twin of Enterococcus faecium metabolism reproduces high-throughput phenotyping data

doi: 10.64898/2026.05.01.720924

Figure Lengend Snippet: Amino acid auxotrophy experiments of E. faecium DO. (A) Mean final optical density (OD) of E. faecium DO in the absence of single amino acids from the CDM-LAB. Bar chart showing final mean OD values after 24 hours incubation. Each bar represents the mean OD of three biological replicates ± SD for a condition where single amino acid was omitted. Cultures with a final OD < 0.1 are marked in red; those with a final OD > 0.3 are marked in blue; and ambiguous growth (OD between 0.1 and 0.3) is color-coded in green. A repeated one-way ANOVA test was performed to compare growth (final average OD 600 ) across amino acid omissions. Results were significantly different ( p value = 0.0024, ≤ 0.05). This was followed by Tukey’s multiple comparisons post hoc test. (B) Repeated passaging of cultures grown in the absence of lysine, phenylalanine, and tyrosine, respectively, results in adaptation to omissions. (C) Individual comparison of each amino acid between the experimental results of the amino acid leave-out experiments (EXP) and the simulation results of the model iDR479 (GEM). Purple squares indicate growth, and green squares indicate no-growth

Article Snippet: The strain whose genome was used in this reconstruction is E. faecium TX0016, also known as the DO strain (ATCC BAA-472), isolated from the blood of a patient with infective endocarditis.

Techniques: Incubation, Passaging, Comparison