Journal: Frontiers in Microbiology

Article Title: Anti-Biofilm Activity of a Self-Aggregating Peptide against Streptococcus mutans

doi: 10.3389/fmicb.2017.00488

Figure Lengend Snippet: Anti-biofilm effect of peptide P1 in microplate assay and on sHA discs. (A) Peptide P1 reduces biofilm formation by Streptococcus mutans and oral streptococci in microplate assay. Cultures were incubated with peptide P1 and control peptide sP1 at 0.1 mg/ml final concentration. Biofilm biomass in microplate wells was stained with crystal violet (images below graph). Results are combined from three replicate experiments, and error bars indicate standard error of the mean. ** p < 0.01, **** p < 0.0001, Student’s t -test. (B) Determination of minimum biofilm inhibitory concentration of peptide P1 against S. mutans . Biofilm inhibitory activity of P1 was measured by testing decreasing peptide concentrations in a microplate assay. Results were combined from three experiments, with biofilm biomass normalized to a percentage of the control. (C) Viability of S. mutans in the presence of peptide P1. Total S. mutans biomass (combined planktonic and biofilm cells from well) of cultures grown overnight in Brain Heart Infusion + 1% sucrose (BHI-S) without peptide or with 0.1 mg/ml P1 or sP1 was collected, diluted and plated, and cfu/ml was quantified for each condition. (D) Peptide P1 reduces Streptococcus mutans biofilm formation on saliva-coated hydroxyapatite (sHA) discs. Biofilm cfu/sHA disc showing combined results from three replicate experiments (total n = 9 discs/treatment). Peptide P1 and control peptide sP1 were used at 0.1 mg/ml final concentration. Error bars show standard error of the mean. * p < 0.05, using Student’s t -test. (E) Custom-made wire holder holding the hydroxyapatite disc vertically in the well of a 96-well plate.

Article Snippet: Streptococci bacteria ( Streptococcus mutans ATCC 25175, Streptococcus oralis ATCC 9811, and Streptococcus salivarius ATCC 13419), Gram-negative bacteria ( Alcaligenes faecalis ATCC 8750, Enterobacter cloacae ATCC 23355, and Salmonella typhimurium ATCC 29629), and Gram-positive bacterial strains ( ) were obtained from Yale University or Presque Isle Cultures (Erie, PA, USA).

Techniques: Incubation, Control, Concentration Assay, Staining, Activity Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Neuraminidase-producing oral mitis group streptococci potentially contribute to influenza viral infection and reduction in antiviral efficacy of zanamivir

doi: 10.1007/s00018-014-1669-1

Figure Lengend Snippet: Effects of bacterial culture supernatants on the release of influenza virus. a Stimulation of influenza virus release by bacterial culture supernatants of oral mitis group streptococci. MDCK cells were infected with A/Udorn/72 influenza virus at an MOI of 0.001. After viral adsorption for 30 min, the cells were incubated with BHI (control) or bacterial culture supernatants (1 or 5 %, v/v). Following 24-h incubation, the culture media were harvested, and virus titers were determined by plaque assays (black bar). NA activities in bacterial culture supernatants were determined using 4-MUNANA (white bar). Values are presented as the mean ± SD, n = 3 (*P < 0.05; **P < 0.01). b Culture supernatants of S. oralis ATCC10557 and S. mitis ATCC6249 promoted the expression of viral M1 proteins. MDCK cells were infected with A/Udorn/72 influenza virus at an MOI of 0.001. After viral adsorption for 30 min, the cells were incubated with BHI (control) or bacterial culture supernatants (5 %, v/v). Following 16-h incubation, the culture supernatants were harvested, and the expression of viral M1 proteins was detected by Western blotting using a specific monoclonal antibody

Article Snippet: Bacterial culture supernatant preparation Actinomyces spp. (WVU 398A, ATCC 12104, ATCC 19039, LY7, ATCC 27044, 8A06, WVU627, T14V, ATCC 19246), Streptococcus oralis (JCM 12997, ATCC 9811, ATCC 10557), Streptococcus mitis (JCM 12971, ATCC 6249) and Streptococcus sanguinis (ATCC 10556) were grown in brain heart infusion (BHI) broth (BD, Franklin Lakes, NJ, USA) at 37 °C for 24 h. P. gingivalis (ATCC 33277, W83, FDC 381, W50, SU 63) were grown in BHI broth containing 5 μg/mL haemin and 0.5 μg/mL menadione in an anaerobic system (10 % CO 2 , 10 % H 2 and 80 % N 2 ; Model 1024, Forma Scientific, Marietta, OH, USA).

Techniques: Virus, Infection, Adsorption, Incubation, Control, Expressing, Western Blot

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Neuraminidase-producing oral mitis group streptococci potentially contribute to influenza viral infection and reduction in antiviral efficacy of zanamivir

doi: 10.1007/s00018-014-1669-1

Figure Lengend Snippet: Culture supernatants of S. mitis ATCC6249 and S. oralis ATCC10557 promoted the cell-to-cell spread of influenza viral infection. a, b NA-producing oral bacteria promoted the spread of influenza virus infection. For indirect immunofluorescence assay, MDCK cells were infected with A/Udorn/72 influenza virus at an MOI of 0.01. After viral adsorption for 1 h, the cells were incubated for 12 h with BHI or bacterial culture supernatants (5 %, v/v). The cells were then fixed and the antigens were stained with rabbit polyclonal antibody against purified A/Udorn/72 virions. Nuclei were counterstained by Hoechst 33342. Depicted is the merged image of virus staining (red) and nuclear DNA counterstaining (blue). The numbers of antigen-positive cells is shown in (b). Values are presented as mean ± SD; n = 3 (*P < 0.05; **P < 0.01). c Effects of bacterial culture supernatants on the expression of viral M1 proteins in the infected cells. Experiments were performed in a manner similar to that described in the legend for Fig. 2b. Cell lysates were harvested, and the expression of viral M1 proteins was detected

Article Snippet: Bacterial culture supernatant preparation Actinomyces spp. (WVU 398A, ATCC 12104, ATCC 19039, LY7, ATCC 27044, 8A06, WVU627, T14V, ATCC 19246), Streptococcus oralis (JCM 12997, ATCC 9811, ATCC 10557), Streptococcus mitis (JCM 12971, ATCC 6249) and Streptococcus sanguinis (ATCC 10556) were grown in brain heart infusion (BHI) broth (BD, Franklin Lakes, NJ, USA) at 37 °C for 24 h. P. gingivalis (ATCC 33277, W83, FDC 381, W50, SU 63) were grown in BHI broth containing 5 μg/mL haemin and 0.5 μg/mL menadione in an anaerobic system (10 % CO 2 , 10 % H 2 and 80 % N 2 ; Model 1024, Forma Scientific, Marietta, OH, USA).

Techniques: Infection, Bacteria, Virus, Immunofluorescence, Adsorption, Incubation, Staining, Purification, Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Neuraminidase-producing oral mitis group streptococci potentially contribute to influenza viral infection and reduction in antiviral efficacy of zanamivir

doi: 10.1007/s00018-014-1669-1

Figure Lengend Snippet: S. mitis ATCC6249 and S. oralis ATCC10557 culture supernatants enhanced ERK activation in the infected cells. MDCK cells were infected with the virus at an MOI of 0.001 in the presence or absence of BHI or bacterial culture supernatants (5 %). After 16-h incubation, cell lysates were harvested and phosphorylated ERK (p-ERK) or total ERK were detected by Western blotting using specific antibodies

Article Snippet: Bacterial culture supernatant preparation Actinomyces spp. (WVU 398A, ATCC 12104, ATCC 19039, LY7, ATCC 27044, 8A06, WVU627, T14V, ATCC 19246), Streptococcus oralis (JCM 12997, ATCC 9811, ATCC 10557), Streptococcus mitis (JCM 12971, ATCC 6249) and Streptococcus sanguinis (ATCC 10556) were grown in brain heart infusion (BHI) broth (BD, Franklin Lakes, NJ, USA) at 37 °C for 24 h. P. gingivalis (ATCC 33277, W83, FDC 381, W50, SU 63) were grown in BHI broth containing 5 μg/mL haemin and 0.5 μg/mL menadione in an anaerobic system (10 % CO 2 , 10 % H 2 and 80 % N 2 ; Model 1024, Forma Scientific, Marietta, OH, USA).

Techniques: Activation Assay, Infection, Virus, Incubation, Western Blot

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Neuraminidase-producing oral mitis group streptococci potentially contribute to influenza viral infection and reduction in antiviral efficacy of zanamivir

doi: 10.1007/s00018-014-1669-1

Figure Lengend Snippet: Effects of culture supernatants of S. mitis ATCC6249 and S. oralis ATCC10557 on the release of influenza virus in the presence of zanamivir. a Zanamivir did not affect bacterial NA activity. NA activities were measured in the absence or presence of zanamivir (250 nM) and are expressed as arbitrary units of fluorescence signals. Values are presented as mean ± SD; n = 3 (**P < 0.01). b NA-producing oral bacteria decreased the antiviral efficacy of zanamivir. MDCK cells were infected with A/Udorn/72 influenza virus at an MOI of 0.001. After viral adsorption for 30 min, the cells treated with zanamivir (250 nM) were incubated with BHI or bacterial culture supernatants (5 %, v/v) for 24 h. The culture media were harvested, and viral titers were determined by plaque assays. Values are presented as mean ± SD; n = 3 (*P < 0.05; **P < 0.01). c, d Culture supernatants of NA-producing oral bacteria reduced the inhibitory effects of zanamivir on the expression of viral M1 proteins and phosphorylation of ERK. Experiments were performed in a manner similar to that described in the legend for b. Culture supernatants (Sup) of viral M1 proteins and lysates of cells (Lysates) were used for detection of M1 proteins by Western blotting (c). The cell lysates were also used to analyze for phosphorylation of ERK (d)

Article Snippet: Bacterial culture supernatant preparation Actinomyces spp. (WVU 398A, ATCC 12104, ATCC 19039, LY7, ATCC 27044, 8A06, WVU627, T14V, ATCC 19246), Streptococcus oralis (JCM 12997, ATCC 9811, ATCC 10557), Streptococcus mitis (JCM 12971, ATCC 6249) and Streptococcus sanguinis (ATCC 10556) were grown in brain heart infusion (BHI) broth (BD, Franklin Lakes, NJ, USA) at 37 °C for 24 h. P. gingivalis (ATCC 33277, W83, FDC 381, W50, SU 63) were grown in BHI broth containing 5 μg/mL haemin and 0.5 μg/mL menadione in an anaerobic system (10 % CO 2 , 10 % H 2 and 80 % N 2 ; Model 1024, Forma Scientific, Marietta, OH, USA).

Techniques: Virus, Activity Assay, Fluorescence, Bacteria, Infection, Adsorption, Incubation, Expressing, Phospho-proteomics, Western Blot

Journal:

Article Title: Purification, Characterization, and Molecular Analysis of the Gene Encoding Glucosyltransferase from Streptococcus oralis

doi:

Figure Lengend Snippet: Effects of anti- S. oralis ATCC 10557 GTase antibody on the GTase activities of various oral streptococci

Article Snippet: For comparison, S. oralis SK23 and ATCC 9811, S. sanguis ATCC 10556, ST3, and ST7, S. gordonii ATCC 10558, F90A, and SK51, S. mitis SK24 and ATCC 903, S. mutans MT8148, S. sobrinus 6715, and S. salivarius HHT were selected from our culture collection.

Techniques: Activity Assay, Inhibition

Journal:

Article Title: Purification, Characterization, and Molecular Analysis of the Gene Encoding Glucosyltransferase from Streptococcus oralis

doi:

Figure Lengend Snippet: Southern blot analysis of chromosomal DNA from various oral streptococci digested with EcoRI. As a probe, the PCR-amplified DNA fragment corresponding to the N-terminal sequence of gtfR was used. Lanes: 1, S. mutans MT8148; 2, S. sobrinus 6715; 3, S. salivarius HHT; 4, S. sanguis ATCC 10556; 5, S. sanguis ST3; 6, S. sanguis ST7; 7, S. oralis ATCC 10557; 8, S. oralis SK23; 9, S. oralis ATCC 9811; 10, S. gordonii ATCC 10558; 11, S. gordonii F90A; 12, S. gordonii SK51; 13, S. mitis SK24; and 14, S. mitis ATCC 903.

Article Snippet: For comparison, S. oralis SK23 and ATCC 9811, S. sanguis ATCC 10556, ST3, and ST7, S. gordonii ATCC 10558, F90A, and SK51, S. mitis SK24 and ATCC 903, S. mutans MT8148, S. sobrinus 6715, and S. salivarius HHT were selected from our culture collection.

Techniques: Southern Blot, Amplification, Sequencing

Journal: Molecules

Article Title: Molecular Mechanisms of Inhibition of Streptococcus Species by Phytochemicals

doi: 10.3390/molecules21020215

Figure Lengend Snippet: Inhibitory effects of phytochemicals against adhesion, biofilm formation and hydrophobicity.

Article Snippet: Cranberry ( Vaccinium macrocarpon Aiton) , High MW non-dialyzable materials Juice powder 25% concentration, dissolved in water , S. mutans MT 8148R, JC2, Ingbritt, ATCC 10449 S. criceti E49 S. oralis ATCC 10557 S. mitis ATCC 9811 S. gordonii Challis , 100–500 μg/mL Inhibition of biofilm formation , Significant inhibition , [ ] .

Techniques: Concentration Assay, Purification, Incubation, Inhibition, Binding Assay, Activity Assay, Cell Surface Hydrophobicity