Journal: International Journal of Nanomedicine

Article Title: Graphene Oxide-Copper Nanocomposites Suppress Cariogenic Streptococcus mutans Biofilm Formation

doi: 10.2147/ijn.s303521

Figure Lengend Snippet: Figure 1 Antibacterial activity of GO-Cu nanocomposites on cariogenic S. mutans. (A) The growth curves of the S. mutans wild-type strain treated with different concentrations of GO-Cu nanocomposites for 24 hours. (B) CFU counting was applied to evaluate the actual antimicrobial effect of GO-Cu nanocomposites. With the treatment of 2.5, 5, 10, 20, 40, and 80 μg/mL GO-Cu nanocomposites for 2 hours, after 10-fold series dilution, S. mutans bacterial suspensions were plated onto Petri dishes, respectively. Viability was calculated by the following formula: viability % = counts of samples incubation with treatment/counts of samples without treatment. Data are shown as mean ± SD *p < 0.05, ***p < 0.01, ****p < 0.001 (vs control).

Article Snippet: All primers designed using MacVector 7.0 software were purchased commercially (Sangon Biotech, Shanghai, China) and were based on the sequence of the S. mutans UA159 (ATCC 700610) genome.21 The strains were grown in brain heart infusion (BHI) medium (Oxoid, Basingstoke, UK) in an atmosphere of 80% N2 and 20% CO2 at 37°C.

Techniques: Activity Assay, Incubation, Control

Journal: International Journal of Nanomedicine

Article Title: Graphene Oxide-Copper Nanocomposites Suppress Cariogenic Streptococcus mutans Biofilm Formation

doi: 10.2147/ijn.s303521

Figure Lengend Snippet: Figure 3 Biofilm disruption by the low-dose GO-Cu nanocomposites. (A) Biomass of 24-hour S. mutans biofilms with the treatment of low-dose GO-Cu nanocomposites, GO particles and CuNPs, respectively. (B) Biomass measurement at different time points. Dynamics of biofilm disruption after treatments with low-dose GO-Cu nanocomposites were calculated. (C) Representative live/dead staining images of 24-hour biofilms of S. mutans between the control group and GO-Cu nanocomposites group (10 μg/mL). Live bacteria were stained green by SYTO 9, and dead bacteria were stained red by PI. Data are shown as mean ± SD *p < 0.05 (vs control). Abbreviation: ns, not significant.

Article Snippet: All primers designed using MacVector 7.0 software were purchased commercially (Sangon Biotech, Shanghai, China) and were based on the sequence of the S. mutans UA159 (ATCC 700610) genome.21 The strains were grown in brain heart infusion (BHI) medium (Oxoid, Basingstoke, UK) in an atmosphere of 80% N2 and 20% CO2 at 37°C.

Techniques: Disruption, Staining, Control, Bacteria

Journal: International Journal of Nanomedicine

Article Title: Graphene Oxide-Copper Nanocomposites Suppress Cariogenic Streptococcus mutans Biofilm Formation

doi: 10.2147/ijn.s303521

Figure Lengend Snippet: Figure 4 Exopolysaccharides production and distribution with in biofilm treated with low-dose GO-Cu nanocomposites. (A) Representative three-dimensional reconstruc tions of 24-hour biofilms of S. mutans between the control group and GO-Cu nanocomposites group. Bacterial cells were stained with SYTO 9 (in green) and exopolysaccharides were labeled with Alexa Fluor 647 (in red), scale bars, 50 μm. Images were taken at 63× magnification. (B) Exopolysachharide/bacteria biomass using Imaris 7.0.0 and Image Pro Plus (*p < 0.05). (C) Amount of WSG and WIG measured using the anthrone method (*p < 0.05).

Article Snippet: All primers designed using MacVector 7.0 software were purchased commercially (Sangon Biotech, Shanghai, China) and were based on the sequence of the S. mutans UA159 (ATCC 700610) genome.21 The strains were grown in brain heart infusion (BHI) medium (Oxoid, Basingstoke, UK) in an atmosphere of 80% N2 and 20% CO2 at 37°C.

Techniques: Control, Staining, Labeling, Bacteria

Journal: International Journal of Nanomedicine

Article Title: Graphene Oxide-Copper Nanocomposites Suppress Cariogenic Streptococcus mutans Biofilm Formation

doi: 10.2147/ijn.s303521

Figure Lengend Snippet: Figure 6 Probing the regulatory landscape of GO-Cu nanocomposites on S. mutans. (A) The Pearson correlation coefficients analyses of each group. (B) The results of Volcano analysis. 5 genes up-regulation and 21 genes down-regulation in the GO-Cu_vs_WT comparison group. (C) The results of RT-qPCR. The expression of copZ, copY and mreD in the GO-Cu_vs_WT comparison group. (D) Details information about the annotated DEGs according transcriptomes analysis. (E) Functional annotation of DEGs based on GO categorization. (F) KEGG pathway enrichment analysis. The longitudinal axis represents the functional annotation information, and the horizontal axis represents the size of the rich factor. The size of qvalue is represented by the color of the point, and the number of differential genes contained under each function is represented by the size of the point. *p < 0.05.

Article Snippet: All primers designed using MacVector 7.0 software were purchased commercially (Sangon Biotech, Shanghai, China) and were based on the sequence of the S. mutans UA159 (ATCC 700610) genome.21 The strains were grown in brain heart infusion (BHI) medium (Oxoid, Basingstoke, UK) in an atmosphere of 80% N2 and 20% CO2 at 37°C.

Techniques: Comparison, Quantitative RT-PCR, Expressing, Functional Assay

Journal: International Journal of Nanomedicine

Article Title: Graphene Oxide-Copper Nanocomposites Suppress Cariogenic Streptococcus mutans Biofilm Formation

doi: 10.2147/ijn.s303521

Figure Lengend Snippet: Figure 7 The suppression of biofilm formation of GO-Cu nanocomposites on cariogenic S. mutans.

Article Snippet: All primers designed using MacVector 7.0 software were purchased commercially (Sangon Biotech, Shanghai, China) and were based on the sequence of the S. mutans UA159 (ATCC 700610) genome.21 The strains were grown in brain heart infusion (BHI) medium (Oxoid, Basingstoke, UK) in an atmosphere of 80% N2 and 20% CO2 at 37°C.

Techniques:

Journal: Journal of Bacteriology

Article Title: Biosynthesis and Transport of the Lantibiotic Mutacin 1140 Produced by Streptococcus mutans

doi: 10.1128/jb.02531-14

Figure Lengend Snippet: Figure 1. Lantibiotic structural elements. A) Covalent structures for nisin and mutacin 1140 with 547 the lanthionine rings labeled from N- to C-terminus. B) Leader sequence alignments of 548 structurally related Class I lantibiotics; i.e. nisin produced by Lactococcus lactis, subtilin 549 produced by Bacillus subtilis, epidermin produced by Staphylococcus epidermidis, gallidermin 550 produced by Staphylococcus gallinarium, Pep5 produced by Staphylococcus epidermidis, 551 epilancin K7 produced by Staphylococcus epidermidis, mutacin Ny266 produced by S. mutans 552 Ny266, mutacin III produced by S. mutans UA787, mutacin 1140 produced by S. mutans 553 JH1140, and for comparison a class II lantibiotic lacticin 481 produced by Lactococcus lactis 554 (16, 18, 47-53). The F(N/D)LD box and C-terminal proline in class I lantibiotics is underlined 555 and in bold print in the sequences. C) Secondary structure prediction using SOPMA for mutacin 556 1140 leader peptide; h (alpha helix), e (extended strand), c (random coil), t (beta turn). Alpha 557 helical regions are underlined, while random coils are in bold in the leader peptide sequence. 558 559 Figure 2. Identification of structural elements within the mutacin 1140 leader peptide that are 560 important for bioactivity. A) Covalent structure representation of the mutations made on the 561 leader peptide. Bioactivity for leader and core peptide mutants were measured as the percent 562 difference in the zone of inhibition between wild-type and the mutant strains. ∆lanA strain was 563 used as a negative control for bioactivity in all experiments. The change in activity was measured 564 for: B) mutations near the LanP cleavage site, C) mutations within the newly discovered 565 cleavage site, and D) mutations in the transporter lanT and core peptide regions responsible for 566 ring formation. For each mutation, the bioactivity has been compared to the activity of wild-type 567 S. mutans JH1140 strain. All the activities are determined in the absence of trypsin in the top 568 agar of the overlays. Statistical method used was Student t-test and the asterisk signifies 569 statistical significance (p<0.05). 570 571 Figure 3. HPLC chromatograms of crude extracts obtained from modified THyex media 572 inoculated with A) wild-type S. mutans JH1140 B) and S. mutans ∆lanP. There is more than a 573 two-fold increase in isolated product from the S. mutans ∆lanP strain. 574 575 Figure 4. Characterization of the leader peptide ∆(-7-2) mutant. A) Overlay assay using M. 576 luteus as an indicator strain. From top left to bottom right: 5 µL spotted of purified mutacin 1140 577 (10 µg/mL) in acetonitrile:water (1:1); (negative control) 5 µL spotted of acetonitrile:water (1:1); 578 deferred antagonism assay of S. mutans ∆(-7-2); deferred antagonism assay of S. mutans 579 ∆lanP∆(-7-2); 5 µL spotted of trypsin digestion of product from S. mutans ∆(-7-2); and 5 µL 580 spotted of trypsin digestion of product from S. mutans ∆lanP∆(-7-2). B) MALDI-MS data of 581 product from S. mutans ∆(-7-2). C) MALDI-MS data of product from S. mutans ∆lanP∆(-7-2). 582 D) MALDI-MS data of the 2-mercaptoethanol (BME) derivitization of the isolated products 583 from S. mutans ∆(-7-2) strain. 584 585 Figure 5. Transport efficiencies of mutacin 1140 core peptide variants. Alanine substitutions 586 were made to disrupt ring A (C(7)A), ring B (C(11)A), ring C (C(21)A), ring D (S(19)A), rings 587 AB (C(7)A:C(11)A), and rings CD (C(21)A:S(19)A). Three independent extractions of each 588 mutant strain were characterized by RP-HPLC at 220 nm. Peak volumes of the variants of 589 mutacin 1140 fractions were compared to wild-type strain JH1140 and are represented as percent 590 of product relative to wild-type strain. Statistical method used was Student t-test and the asterisk 591 signifies statistical significance (p<0.05). 592 593

Article Snippet: THyex broth (30 g/L Todd Hewitt Broth, 3 g/L yeast extract), THyex agar media 439 (30 g/L Todd Hewitt Broth, 3 g/L yeast extract, 15 g/L agar; Bacto, Sparks, MD) and Top agar 440 media (30 g/L Todd Hewitt Broth, 3 g/L yeast extract, 7.5 g/L agar; Bacto, Sparks, MD) was 441 used to culture S. mutans JH1140 ATCC 55676 and Micrococcus luteus ATCC 10240 at 37 °C.

Techniques: Labeling, Sequencing, Produced, Comparison, Inhibition, Mutagenesis, Negative Control, Activity Assay, Modification, Isolation, Overlay Assay, Purification

Journal: Oral health and dental management

Article Title: Cariogenicity of Streptococcus mutans Glucan-Binding Protein Deletion Mutants

doi:

Figure Lengend Snippet: S. mutans strains used in this study.

Article Snippet: The following strains were used in this study: E. coli JM109 (Promega, Madison, WI, USA); S. mutans UA130 (provided by Dr. Suzanne Michalek, University of Alabama-Birmingham) was used as the wild-type (WT) and parental strain for generation of Gbp mutants; and S. sanguinis ATCC 10556 (provided by Dr. David Drake, University of Iowa) was used in adhesion assays.

Techniques: Mutagenesis

Journal: Biofouling

Article Title: Anti-biofilm activity of silver nanoparticles against different microorganisms.

doi: 10.1080/08927014.2013.794225

Figure Lengend Snippet: Figure 2. The inhibition of the growth of the biofilm using a colony model. (A) Inhibition of the growth of microorganisms with varying concentrations of AgNPs. (B) Selected images showing the inhibition in the formation of biofilms at a 1000 mg ml1 concentration of AgNP. Ab, A. baumannii; Ca, C. albicans; MRSA, methicillin-resistant S. aureus; Pa, P. aeruginosa; Sm, S. mutans.

Article Snippet: As representative Gram-positive microorganisms, the strains methicillinresistant Staphylococcus aureus (MRSA) (ATCC 700787) and Streptococcus mutans (ATCC 33535) were evaluated.

Techniques: Inhibition, Concentration Assay