Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD11b/CD18-dependent interactions of neutrophils with intestinal epithelium are mediated by fucosylated proteoglycans.

doi: 10.4049/jimmunol.169.9.5270

Figure Lengend Snippet: FIGURE 1. Concentration-dependent inhibition of T84 cell adhesion to CD11b/CD18 by fucoidin. As detailed in Materials and Methods, BCECF- labeled T84 cells were incubated with CD11b/CD18-coated wells for 1 h at 37°C in the presence of polysaccharides. T84 cell adhesion is expressed as a percentage of the total applied cells. Values are the mean SD of three separate experiments, each performed in triplicate.

Article Snippet: Hybridoma cells secreting a functionally inhibitory mAb against CD18 (TS1/18, IgG1) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Concentration Assay, Inhibition, Labeling, Incubation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD11b/CD18-dependent interactions of neutrophils with intestinal epithelium are mediated by fucosylated proteoglycans.

doi: 10.4049/jimmunol.169.9.5270

Figure Lengend Snippet: FIGURE 2. Fucoidin-mediated inhibition of T84 cell adhesion to CD11b/CD18 occurs through direct interactions with CD11b/CD18. Be- fore cell adhesion assays, T84 cells (FT84 cells) or CD11b/CD18-coated microtiter wells (FCD11b/CD18) were preincubated with 10 mg/ml of fucoidin for 30 min at 37°C, followed by washing. Cell adhesion assays were then performed in the absence of fucoidin as detailed in Materials and Methods. Control wells (no treatment) represent T84 cell binding to CD11b/CD18 in the absence of fucoidin pretreatment, whereas blank rep- resents binding of T84 cells to wells coated with BSA only. Values are the mean SD of three separate experiments, each performed in triplicate.

Article Snippet: Hybridoma cells secreting a functionally inhibitory mAb against CD18 (TS1/18, IgG1) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Inhibition, Control, Binding Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD11b/CD18-dependent interactions of neutrophils with intestinal epithelium are mediated by fucosylated proteoglycans.

doi: 10.4049/jimmunol.169.9.5270

Figure Lengend Snippet: FIGURE 3. Binding of fucoidin to CD11b/CD18. A, Binding of biotin- ylated fucoidin (B-fucoidin) to immobilized CD11b. As detailed in Mate- rials and Methods, microtiter wells coated with CD11b/CD18 were blocked with BSA, followed by addition of 100 g/ml B-fucoidin for 1 h at 37°C. B-fucoidin binding was assessed after addition of HRP-streptavi- din, followed by color development and OD measurement (405 nm). B, Binding of CD11b/CD18 to immobilized fucoidin and chondroitin sulfate C (control). Polysaccharides (5 mg/ml in HBSS) were added to wells (50 l/well, 37°C, 2 h), followed by blocking with BSA and addition of CD11b/CD18 (2.5 g/ml, 37°C, 1 h). CD11b/CD18 binding to fucoidin was assessed after addition of mAb TS1/18, followed by incubation with HRP-conjugated secondary mAb, color development, and OD measure- ment (405 nm). Binding assays were performed in the presence of 2 mM Ca2 and Mg2. BSA-coated wells served as controls in both experiments. Values are the mean SD of three separate experiments, each performed in triplicate.

Article Snippet: Hybridoma cells secreting a functionally inhibitory mAb against CD18 (TS1/18, IgG1) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Binding Assay, Control, Blocking Assay, Incubation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD11b/CD18-dependent interactions of neutrophils with intestinal epithelium are mediated by fucosylated proteoglycans.

doi: 10.4049/jimmunol.169.9.5270

Figure Lengend Snippet: FIGURE 4. Effects of anti-CD11b/CD18 mAbs and EDTA on the bind- ing of fucoidin to CD11b/CD18. Binding assays of biotinylated fucoidin (B-fucoidin, 100 g/ml) to immobilized CD11b/CD18 were performed as detailed in Fig. 3, with the inclusion of CD11b/CD18 mAbs (40 g/ml) or 5 mM EDTA. Values are the mean SD of three separate experiments, each performed in triplicate.

Article Snippet: Hybridoma cells secreting a functionally inhibitory mAb against CD18 (TS1/18, IgG1) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Binding Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD11b/CD18-dependent interactions of neutrophils with intestinal epithelium are mediated by fucosylated proteoglycans.

doi: 10.4049/jimmunol.169.9.5270

Figure Lengend Snippet: FIGURE 5. Fucoidin neoproteoglycan binds to CD11b/CD18. As de- tailed in Materials and Methods, a neoproteoglycan derivative of fucoidin (F-BSA) was prepared for this experiment. Purified F-BSA (5 g/lane; lane 1) and BSA (lane 2) were separated by nonreducing SDS-PAGE (4– 15% gel). The proteins were either visualized by silver staining (A) or transferred to nitrocellulose filters and probed with purified CD11b/CD18 (B), followed by incubation with mAb TS1/18 and HRP-conjugated sec- ondary mAb, and visualized by ECL. All buffers contained 2 mM Mg2.

Article Snippet: Hybridoma cells secreting a functionally inhibitory mAb against CD18 (TS1/18, IgG1) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: SDS Page, Silver Staining, Incubation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD11b/CD18-dependent interactions of neutrophils with intestinal epithelium are mediated by fucosylated proteoglycans.

doi: 10.4049/jimmunol.169.9.5270

Figure Lengend Snippet: FIGURE 6. Fucoidin binding to CD11b/CD18 is dependent on sulfation of polysaccharide. A, Binding of fucoidin or desulfated-fucoidin (De-fu- coidin) to immobilized CD11b/CD18. Fucoidin was biotinylated and des- ulfated as detailed in Materials and Methods. Biotinylated polysaccharide (100 g/ml) was incubated with CD11b/CD18-coated wells or blanks (no CD11b/CD18) for 1 h at 37°C, followed by addition of HRP-streptavidin and color development. B, Effect of fucoidin or De-fucoidin on T84 cell adhesion to CD11b/CD18. Suspensions of BCECF-labeled T84 cells (50 l, 106 cells/ml) were incubated with CD11b/CD18-coated wells or blanks (no CD11b/CD18) for 1 h at 37°C in the presence or the absence of 10 mg/ml polysaccharides as indicated. The control represents binding of T84 cells to CD11b/CD18 in the absence of polysaccharide. Values are the mean SD of three separate experiments, each performed in triplicate.

Article Snippet: Hybridoma cells secreting a functionally inhibitory mAb against CD18 (TS1/18, IgG1) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Binding Assay, Incubation, Labeling, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD11b/CD18-dependent interactions of neutrophils with intestinal epithelium are mediated by fucosylated proteoglycans.

doi: 10.4049/jimmunol.169.9.5270

Figure Lengend Snippet: FIGURE 7. Effect of fucosidase on epithelial cell adherence to CD11b/CD18. As detailed in Materials and Methods, trypsin/EDTA-elicited T84 cells were pretreated with fucosidase or neuraminidase (0, 0.1, 0.5, and 1.0 U/ml, respectively) for 30 min at 37°C, washed twice with HBSS and used in adhesion assays (A). B, Cells were pretreated with fucosidase (0.5 U/ml) or neuraminidase (0.5 U/ml) at 37°C for different lengths of time as indicated, followed by washing with HBSS, and were used in CD11b/CD18 adhesion assays. Values are the mean SD of three separate experiments, each performed in triplicate.

Article Snippet: Hybridoma cells secreting a functionally inhibitory mAb against CD18 (TS1/18, IgG1) were purchased from American Type Culture Collection (Manassas, VA).

Techniques:

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD11b/CD18-dependent interactions of neutrophils with intestinal epithelium are mediated by fucosylated proteoglycans.

doi: 10.4049/jimmunol.169.9.5270

Figure Lengend Snippet: FIGURE 9. Inhibition of T84 cell proteoglycan synthesis reduces cell adhesion to CD11b/CD18. As detailed in Materials and Methods, T84 cells were cultured in the presence of 2 mM -xyl or 2 mM -xyl in DMEM overnight, followed by harvest and BCECF labeling for cell adhesion as- says. In a subset of experiments T84 cells pretreated with -xyl were fur- ther incubated with fucosidase (1.0 U/ml) for 30 min at 37°C before ad- hesion assays. Values are the mean SD of three separate experiments, each performed in triplicate. , p 0.01.

Article Snippet: Hybridoma cells secreting a functionally inhibitory mAb against CD18 (TS1/18, IgG1) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Inhibition, Cell Culture, Labeling, Incubation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD11b/CD18-dependent interactions of neutrophils with intestinal epithelium are mediated by fucosylated proteoglycans.

doi: 10.4049/jimmunol.169.9.5270

Figure Lengend Snippet: FIGURE 10. Identification of fucosylated CD11b/CD18 binding pro- teins in T84 cells. Fucosylated proteins were isolated from T84 cell lysate via a fucose-specific lectin column as detailed in Materials and Methods. Fucosylated proteins were analyzed by 12% SDS-PAGE (10 g/lane) and stained with GelCode Blue (lane 1). In parallel, proteins were transferred to nitrocellulose filters and probed with CD11b/CD18 (lane 3), followed by incubation with mAb TS1/18 and HRP-conjugated secondary mAb, and were visualized by ECL. All buffers in this experiment contained 2 mM Mg2. Lane 2, Control nitrocellulose filter performed as described for lane 3, but without incubation with CD11b/CD18.

Article Snippet: Hybridoma cells secreting a functionally inhibitory mAb against CD18 (TS1/18, IgG1) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Binding Assay, Isolation, SDS Page, Staining, Incubation, Control

Journal: International Journal of Pharmaceutics: X

Article Title: Nanostructured N/S doped carbon dots/mesoporous silica nanoparticles and PVA composite hydrogel fabrication for anti-microbial and anti-biofilm application

doi: 10.1016/j.ijpx.2023.100209

Figure Lengend Snippet: Antimicrobial activity test of CDs, NS/CDs, CDs@MSN, NS/CDs@MSN toward the growth of Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 27853).

Article Snippet: For synthesized CDs and CDs@MSN nanoparticles, antimicrobial susceptibility study against the growth of Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 27853) was studied using a disk diffusion method.

Techniques: Activity Assay, Inhibition

Journal: International Journal of Pharmaceutics: X

Article Title: Nanostructured N/S doped carbon dots/mesoporous silica nanoparticles and PVA composite hydrogel fabrication for anti-microbial and anti-biofilm application

doi: 10.1016/j.ijpx.2023.100209

Figure Lengend Snippet: Effect of nanocomposites on biofilm inhibition activity against Staphylococcus aureus.

Article Snippet: For synthesized CDs and CDs@MSN nanoparticles, antimicrobial susceptibility study against the growth of Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 27853) was studied using a disk diffusion method.

Techniques: Inhibition, Activity Assay, Concentration Assay

Journal: International Journal of Pharmaceutics: X

Article Title: Nanostructured N/S doped carbon dots/mesoporous silica nanoparticles and PVA composite hydrogel fabrication for anti-microbial and anti-biofilm application

doi: 10.1016/j.ijpx.2023.100209

Figure Lengend Snippet: Influence of different PVA hydrogels on antimicrobial and antibiofilm forming activity against Staphylococcus aureus.

Article Snippet: For synthesized CDs and CDs@MSN nanoparticles, antimicrobial susceptibility study against the growth of Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 27853) was studied using a disk diffusion method.

Techniques: Activity Assay