Journal: bioRxiv

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1101/551168

Figure Lengend Snippet: Commensal strains were cultured in BHI till the optical density at 600 nm (OD 600 ) reached 0.5. Cultures were then diluted 1:50 into FMC medium supplemented with 10 mM Glc (red), GlcN (blue), or GlcNAc (green). The OD 600 was monitored using a Bioscreen C with readings taken every 30 min.

Article Snippet: Commensals were cultured overnight in BHI and sub-cultured in BHI broth to mid-exponential phase (OD 600 ≈ 0.5), then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN) or N-Acetyl-D-glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques: Cell Culture

Journal: bioRxiv

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1101/551168

Figure Lengend Snippet: Each commensal was mixed with UA159-Km in equal proportions and inoculated into BM supplemented with 18 mM Glc and 2 mM sucrose (BMGS) to form biofilms overnight on glass coverslips. BMGS was then replaced by BM supplemented with 20 mM Glc, GlcN, or GlcNAc and incubated for another 24 h before (A) visualization by confocal laser scanning microscopy (CLSM) after LIVE/DEAD staining and (B) CFU quantification of both species. Biofilms formed by UA159-Km alone were included for comparison. Values show the percentages of the populations constituted by commensal (top) and S. mutans (bottom) respectively. Asterisks indicate statistically significant differences in the viable counts of S. mutans compared to that on BMGlc, * p <0.05, ** p <0.01, *** p <0.001, **** p <0.001.

Article Snippet: Commensals were cultured overnight in BHI and sub-cultured in BHI broth to mid-exponential phase (OD 600 ≈ 0.5), then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN) or N-Acetyl-D-glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques: Incubation, Confocal Laser Scanning Microscopy, Staining

Journal: bioRxiv

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1101/551168

Figure Lengend Snippet: Cells were cultivated to early exponential phase in TY supplemented with 20 mM Glc, GlcN or GlcNAc, exposed to vigorous aeration and the concentrations of H 2 O 2 in the supernates were determined as described in the methods section. Asterisks indicate statistically significant differences in H 2 O 2 levels relative to those on glucose, * p <0.05, ** p <0.01, *** p <0.001, **** p <0.001.

Article Snippet: Commensals were cultured overnight in BHI and sub-cultured in BHI broth to mid-exponential phase (OD 600 ≈ 0.5), then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN) or N-Acetyl-D-glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques:

Journal: bioRxiv

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1101/551168

Figure Lengend Snippet: Commensals and S. mutans UA159 were spotted on TY agar plates containing 20 mM Glc, GlcN, or GlcNAc, and incubated under aerobic or anaerobic conditions. The competitions were initiated with colonization by UA159 ( S. m first), or commensals (Commensal first) alone for 24 h then followed by the other, or by both (Simultaneously).

Article Snippet: Commensals were cultured overnight in BHI and sub-cultured in BHI broth to mid-exponential phase (OD 600 ≈ 0.5), then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN) or N-Acetyl-D-glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques: Incubation

Journal: bioRxiv

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1101/551168

Figure Lengend Snippet: For AD assays, commensals were grown to exponential phase in liquid TY supplemented with 20 mM Glc, GlcN, GlcNAc, or galactose, in addition to 10 mM arginine. Asterisks denote statistically significant difference compared to cultures prepared with glucose, * p <0.05, ** p <0.01, *** p <0.001, **** p <0.001.

Article Snippet: Commensals were cultured overnight in BHI and sub-cultured in BHI broth to mid-exponential phase (OD 600 ≈ 0.5), then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN) or N-Acetyl-D-glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques:

Journal: bioRxiv

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1101/551168

Figure Lengend Snippet: Washed cells of each clinical commensal were mixed with UA159 at a 1:1 ratio based on optical density and stabbed onto TY agar plates containing 20 mM Glc, GlcN, or GlcNAc. The plates were incubated overnight, overlaid with soft agar containing S. sanguinis SK150, and then incubated for another 24 h. The experiments were carried out in both aerobic and anaerobic environments.

Article Snippet: Commensals were cultured overnight in BHI and sub-cultured in BHI broth to mid-exponential phase (OD 600 ≈ 0.5), then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN) or N-Acetyl-D-glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques: Incubation

Journal: Applied and Environmental Microbiology

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1128/AEM.00370-19

Figure Lengend Snippet: Growth curves. Commensal strains were cultured in BHI until the OD600 reached 0.5. Cultures were then diluted 1:50 into FMC medium supplemented with 10 mM Glc (red), GlcN (blue), or GlcNAc (green). The OD600 was monitored using a Bioscreen C with readings taken every 30 min.

Article Snippet: Commensals were cultured overnight in BHI and subcultured in BHI broth to mid-exponential phase (optical density at 600 nm [OD 600 ] of ≈0.5) and then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN), or N -acetyl- d -glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques: Cell Culture

Journal: Applied and Environmental Microbiology

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1128/AEM.00370-19

Figure Lengend Snippet: Dual-species biofilms formed using clinical commensals and S. mutans. Each commensal was mixed with UA159-Km in equal proportions and inoculated into BM supplemented with 18 mM Glc and 2 mM sucrose (BMGS) to form biofilms overnight on glass coverslips. BMGS was then replaced by BM supplemented with 20 mM Glc, GlcN, or GlcNAc and incubated for another 24 h before visualization by confocal laser scanning microscopy (CLSM) after LIVE/DEAD staining (A) and CFU quantification of both species (B). Biofilms formed by UA159-Km alone were included for comparison. Values show the percentages of the populations constituted by commensal bacteria (top) and S. mutans (bottom), respectively. Asterisks indicate statistically significant differences in the viable counts of S. mutans compared to that on BMGlc. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Commensals were cultured overnight in BHI and subcultured in BHI broth to mid-exponential phase (optical density at 600 nm [OD 600 ] of ≈0.5) and then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN), or N -acetyl- d -glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques: Incubation, Confocal Laser Scanning Microscopy, Staining

Journal: Applied and Environmental Microbiology

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1128/AEM.00370-19

Figure Lengend Snippet: H2O2 production. Cells were cultivated to early exponential phase in TY supplemented with 20 mM Glc, GlcN, or GlcNAc and exposed to vigorous aeration, and the concentrations of H2O2 in the supernates were determined as described in Materials and Methods. Asterisks indicate statistically significant differences in H2O2 levels relative to those on glucose. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Commensals were cultured overnight in BHI and subcultured in BHI broth to mid-exponential phase (optical density at 600 nm [OD 600 ] of ≈0.5) and then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN), or N -acetyl- d -glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques:

Journal: Applied and Environmental Microbiology

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1128/AEM.00370-19

Figure Lengend Snippet: Competition assays between S. mutans and clinical commensals. Commensals and S. mutans UA159 were spotted on TY agar plates containing 20 mM Glc, GlcN, or GlcNAc and incubated under aerobic or anaerobic conditions. The competitions were initiated with colonization by UA159 (S. mutants first) or by commensals (commensal first) alone for 24 h and then followed by the other or by both (simultaneously).

Article Snippet: Commensals were cultured overnight in BHI and subcultured in BHI broth to mid-exponential phase (optical density at 600 nm [OD 600 ] of ≈0.5) and then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN), or N -acetyl- d -glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques: Incubation

Journal: Applied and Environmental Microbiology

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1128/AEM.00370-19

Figure Lengend Snippet: Arginine deiminase (AD) activity. For AD assays, commensals were grown to exponential phase in liquid TY supplemented with 20 mM Glc, GlcN, GlcNAc, or galactose, in addition to 10 mM arginine. Asterisks denote statistically significant difference compared to cultures prepared with glucose. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Commensals were cultured overnight in BHI and subcultured in BHI broth to mid-exponential phase (optical density at 600 nm [OD 600 ] of ≈0.5) and then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN), or N -acetyl- d -glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques: Activity Assay

Journal: Applied and Environmental Microbiology

Article Title: Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans

doi: 10.1128/AEM.00370-19

Figure Lengend Snippet: Plate-based bacteriocin assay. Washed cells of each clinical commensal were mixed with UA159 at a 1:1 ratio based on optical density and stabbed onto TY agar plates containing 20 mM Glc, GlcN, or GlcNAc. The plates were incubated overnight, overlaid with soft agar containing S. sanguinis SK150, and then incubated for another 24 h. The experiments were carried out in both aerobic and anaerobic environments.

Article Snippet: Commensals were cultured overnight in BHI and subcultured in BHI broth to mid-exponential phase (optical density at 600 nm [OD 600 ] of ≈0.5) and then diluted 1:50 into modified FMC medium formulated with final concentrations of 10 mM glucose (Glc), glucosamine (GlcN), or N -acetyl- d -glucosamine (GlcNAc) (Sigma Millipore, St. Louis, MO).

Techniques: Incubation