Journal: Frontiers in Microbiology

Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation

doi: 10.3389/fmicb.2018.02130

Figure Lengend Snippet: The schematic diagram of the mecA flanking region (A) and RT-PCR analysis (B) . (A) Schematic diagram of the flanking regions of mecA in the genome of S. mutans UA159. Numbers underneath represent the size of the respective genes. Arrows above and underneath indicate the location and orientation of the primers used for reverse transcription (5) and cDNA amplification (F and R). (B) RT-PCR analysis of the mecA / rgpG cluster. cDNA generated from reverse transcription using total RNA extract of S. mutans was PCR amplified using primer set PmecARgpG (lane 1), which shows a single amplicon indicative of co-transcription of mecA and rgpG . Negative control (lane 2) with no reverse transcriptase shows no DNA amplification. M, for molecular marker.

Article Snippet: For immunoblot analysis, equal amount of the above antigen preps were blotted onto a nitrocellulose membrane using a Slot Blot (Bio-Dot SF, BioRad), and probed with S. mutans whole cell antiserum that was generated using inactivated whole cells of the wild-type, UA159 to immunize rabbits (Lampire Biological Laboratories, Inc.) and was adsorbed with live cells of either the wild-type or the rgpG/brpA/psr triple mutant ( ).

Techniques: Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, Amplification, Generated, Negative Control, DNA Amplification, Marker

Journal: Frontiers in Microbiology

Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation

doi: 10.3389/fmicb.2018.02130

Figure Lengend Snippet: Colony morphology of the mecA mutant. S. mutans wild-type (UA159), the mecA mutant (TW416) and its complement strain (TW416C) were grown on BHI agar plates. Relative to the rough, dry colonies of UA159, the colonies of TW416 were round, mucoid, and smooth. Images were taken using Samsung Galaxy Note V.

Article Snippet: For immunoblot analysis, equal amount of the above antigen preps were blotted onto a nitrocellulose membrane using a Slot Blot (Bio-Dot SF, BioRad), and probed with S. mutans whole cell antiserum that was generated using inactivated whole cells of the wild-type, UA159 to immunize rabbits (Lampire Biological Laboratories, Inc.) and was adsorbed with live cells of either the wild-type or the rgpG/brpA/psr triple mutant ( ).

Techniques: Mutagenesis

Journal: Frontiers in Microbiology

Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation

doi: 10.3389/fmicb.2018.02130

Figure Lengend Snippet: Growth characterization of the mecA mutant. (A) Growth phenotypes of S. mutans wild-type (UA159), the mecA mutant (TW416) and its complement strain (TW416C) when grown in BHI broth overnight. Arrow indicates severe clumping of the mecA mutant at the tip of the tooth pick. (B) S. mutans strains grown in BHI broth, BHI broth with pH adjusted to 6.0 and BHI broth with inclusion of methyl viologen at 12.5 mM. The optical densities of the cultures at 600 nm were recorded continuously using a Bioscreen C. Data presented in panel (B) are representatives of three separate experiments.

Article Snippet: For immunoblot analysis, equal amount of the above antigen preps were blotted onto a nitrocellulose membrane using a Slot Blot (Bio-Dot SF, BioRad), and probed with S. mutans whole cell antiserum that was generated using inactivated whole cells of the wild-type, UA159 to immunize rabbits (Lampire Biological Laboratories, Inc.) and was adsorbed with live cells of either the wild-type or the rgpG/brpA/psr triple mutant ( ).

Techniques: Mutagenesis

Journal: Frontiers in Microbiology

Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation

doi: 10.3389/fmicb.2018.02130

Figure Lengend Snippet: TEM analysis. S. mutans wildtype (UA159 and A&D), the mecA mutant (TW416 and B&E) and its mecA complement strain (TW416C and C&F) were grown in BHI broth until mid-exponential phase (OD 600nm ≈0.4). (A,B,C) were images taken at magnification of 10,000 × g , and (D,EF) at 25,000×, respectively. Scale bars represent 500 nm. Images in (G) are inserts of blow-up regions of the cell envelope of the different strains with the mutant showing a fuzzy, loose cell envelope (ce) and a thin cytoplasmic membrane (cm), as indicated. The arrows in (E) indicate the fuzzy, loose cell envelope and the asterisks indicating low-density patches of the deficient mutant.

Article Snippet: For immunoblot analysis, equal amount of the above antigen preps were blotted onto a nitrocellulose membrane using a Slot Blot (Bio-Dot SF, BioRad), and probed with S. mutans whole cell antiserum that was generated using inactivated whole cells of the wild-type, UA159 to immunize rabbits (Lampire Biological Laboratories, Inc.) and was adsorbed with live cells of either the wild-type or the rgpG/brpA/psr triple mutant ( ).

Techniques: Mutagenesis, Membrane

Journal: Frontiers in Microbiology

Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation

doi: 10.3389/fmicb.2018.02130

Figure Lengend Snippet: Biofilm formation. S. mutans wildtype (UA159), the mecA mutant (TW416) and its complement strain (TW416C) were grown in BM medium with glucose and sucrose (BMGS), glucose (BMG) or sucrose (BMS). Biofilms were grown on polystyrene surface in 96 well plates and analyzed using a spectrophotometer. Results presented here represent mean absorbance at 575 nm (±standard deviation in error bars) from three independent experiments and ∗ and # P < 0.01, P < 0.05, respectively.

Article Snippet: For immunoblot analysis, equal amount of the above antigen preps were blotted onto a nitrocellulose membrane using a Slot Blot (Bio-Dot SF, BioRad), and probed with S. mutans whole cell antiserum that was generated using inactivated whole cells of the wild-type, UA159 to immunize rabbits (Lampire Biological Laboratories, Inc.) and was adsorbed with live cells of either the wild-type or the rgpG/brpA/psr triple mutant ( ).

Techniques: Mutagenesis, Spectrophotometry, Standard Deviation

Journal: Frontiers in Microbiology

Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation

doi: 10.3389/fmicb.2018.02130

Figure Lengend Snippet: Confocal microscopic analysis of biofilms. S. mutans wildtype (UA159), the mecA mutant (TW416) and its complement strain (TW416C) were grown in BM medium with glucose and sucrose, glucose or sucrose. Biofilms were grown on HA disks vertically placed in 12 well plates for 24 h, and analyzed using a laser scanning confocal microscope. Panel shows representatives of the compressed confocal images at xy, yz, and xz axis of biofilms of UA159, TW416, and TW416C grown in BM plus glucose and sucrose.

Article Snippet: For immunoblot analysis, equal amount of the above antigen preps were blotted onto a nitrocellulose membrane using a Slot Blot (Bio-Dot SF, BioRad), and probed with S. mutans whole cell antiserum that was generated using inactivated whole cells of the wild-type, UA159 to immunize rabbits (Lampire Biological Laboratories, Inc.) and was adsorbed with live cells of either the wild-type or the rgpG/brpA/psr triple mutant ( ).

Techniques: Mutagenesis, Microscopy

Journal: Frontiers in Microbiology

Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation

doi: 10.3389/fmicb.2018.02130

Figure Lengend Snippet: SEM analysis of biofilms. S. mutans wildtype (UA159), the mecA mutant (TW416) and its complement strain (TW416C) were grown in BM medium with glucose and sucrose, glucose or sucrose. Biofilms were grown on HA disks vertically placed in 12 well plates for 24 h, and analyzed using a scanning electron microscope (SEM). Panel shows images of UA159 and TW416 biofilms grown in BM plus glucose and sucrose, which were taken at magnification of 5,000 and 10,000 × g as indicated, with arrows indicating extracellular polymeric substances and asterisks indicating broken cells and cell debris, respectively.

Article Snippet: For immunoblot analysis, equal amount of the above antigen preps were blotted onto a nitrocellulose membrane using a Slot Blot (Bio-Dot SF, BioRad), and probed with S. mutans whole cell antiserum that was generated using inactivated whole cells of the wild-type, UA159 to immunize rabbits (Lampire Biological Laboratories, Inc.) and was adsorbed with live cells of either the wild-type or the rgpG/brpA/psr triple mutant ( ).

Techniques: Mutagenesis, Microscopy

Journal: Frontiers in Microbiology

Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation

doi: 10.3389/fmicb.2018.02130

Figure Lengend Snippet: Acid and hydrogen peroxide challenge assays. S. mutans strains were grown until mid-exponential phase (OD 600nm ≈0.3) and then subjected to an acid or hydrogen peroxide killing. (A) shows survival rate of wildtype (UA159), the mecA mutant (TW416) and its complement strain (TW416C) following incubation in glycin buffer of pH 2.8, while (B) shows survival rate of the strains in the presence of hydrogen peroxide. Data represented here are means (±standard deviation) of at least three independent experiments, with ∗ P < 0.001 as comparing to the wild-type.

Article Snippet: For immunoblot analysis, equal amount of the above antigen preps were blotted onto a nitrocellulose membrane using a Slot Blot (Bio-Dot SF, BioRad), and probed with S. mutans whole cell antiserum that was generated using inactivated whole cells of the wild-type, UA159 to immunize rabbits (Lampire Biological Laboratories, Inc.) and was adsorbed with live cells of either the wild-type or the rgpG/brpA/psr triple mutant ( ).

Techniques: Mutagenesis, Incubation, Standard Deviation

Journal: BMC Oral Health

Article Title: Preventive effects of probiotics on dental caries in vitro and in vivo

doi: 10.1186/s12903-024-04703-x

Figure Lengend Snippet: The experimental protocol in different groups

Article Snippet: S. mutans CCTCC AB 99,010 was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques:

Journal: BMC Oral Health

Article Title: Preventive effects of probiotics on dental caries in vitro and in vivo

doi: 10.1186/s12903-024-04703-x

Figure Lengend Snippet: The amounts of S. mutans in Co-culture condition. * P < 0.05, compared with the control group; ** p < 0.01, compared with the control group

Article Snippet: S. mutans CCTCC AB 99,010 was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Co-Culture Assay, Control

Journal: BMC Oral Health

Article Title: Preventive effects of probiotics on dental caries in vitro and in vivo

doi: 10.1186/s12903-024-04703-x

Figure Lengend Snippet: The amounts of S. mutans in the supernatant, the bacteria and the control groups. * P < 0.05, compared with the control group; ** p < 0.01, compared with the control group

Article Snippet: S. mutans CCTCC AB 99,010 was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Bacteria, Control

Journal: BMC Oral Health

Article Title: Preventive effects of probiotics on dental caries in vitro and in vivo

doi: 10.1186/s12903-024-04703-x

Figure Lengend Snippet: Counts of S. mutans in the oral cavity of each group of rats. * P < 0.05, compared with the control group; ** p < 0.01, compared with the control group; ns, not significant

Article Snippet: S. mutans CCTCC AB 99,010 was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Control

Journal: Frontiers in Microbiology

Article Title: A GntR Family Transcription Factor in Streptococcus mutans Regulates Biofilm Formation and Expression of Multiple Sugar Transporter Genes

doi: 10.3389/fmicb.2018.03224

Figure Lengend Snippet: Deletion of stsR decreases S. mutans biofilm formation at early stage. S. mutans was cultured in BM supplemented with 1% sucrose for 6, 12, 24, and 48 h. The biofilm biomass was determined by CV staining method. Data from three biological replicates were averaged, and the statistical significance between the stsR mutant, wild-type, and complement strain was determined by Student’s t -test. Error bars represent standard deviations based on results from at least three biological replicates. ∗∗ Indicates a significance of P < 0.01.

Article Snippet: According to the National Center for Biotechnology Information (NCBI) S. mutans genome annotation, the function of the majority of the differentially expressed genes (DEGs) are unknown.

Techniques: Cell Culture, Staining, Mutagenesis

Journal: Frontiers in Microbiology

Article Title: A GntR Family Transcription Factor in Streptococcus mutans Regulates Biofilm Formation and Expression of Multiple Sugar Transporter Genes

doi: 10.3389/fmicb.2018.03224

Figure Lengend Snippet: Determination of growth curves of S. mutans UA159, S. mutans Δ stsR , and Δ stsR/pDL278-stsR . S. mutans UA159, S. mutans Δ stsR , and Δ stsR/pDL278-stsR strains were cultivated in BHI to mid-exponential phase and then diluted into (A) fresh BHI broth or (B) TV base medium supplemented with either 10 mM (limiting) or 100 mM (excess) sucrose, glucose, or lactose. Growth curves were monitored with a Multiskan Spectrum (Thermo, Multiskan Go, United States), and the OD 600 was measured in 1 h intervals. (A) For CFU counts, after diluted, the bacteria were cultured at 37°C for 1, 4, 8, and 12 h. Then bacterial suspension was serially diluted in BHI and plated on BHI agar plates. CFU values were calculated after the plates were incubated anaerobically at 37°C for 48 h. Error bars represent standard deviations based on results from at least three biological replicates.

Article Snippet: According to the National Center for Biotechnology Information (NCBI) S. mutans genome annotation, the function of the majority of the differentially expressed genes (DEGs) are unknown.

Techniques: Bacteria, Cell Culture, Suspension, Incubation

Journal: Frontiers in Microbiology

Article Title: A GntR Family Transcription Factor in Streptococcus mutans Regulates Biofilm Formation and Expression of Multiple Sugar Transporter Genes

doi: 10.3389/fmicb.2018.03224

Figure Lengend Snippet: Deletion of stsR decreases the amount of glucans in S. mutans biofilm. The amounts of (A) water insoluble glucans and (B) water soluble glucans in the biofilms of S. mutans UA159, S. mutans Δ stsR , and complement strain were quantified using the phenol-sulfuric acid method and calculated according to the standard curve. Error bars represent standard deviations based on results from at least three biological replicates. ∗∗ Indicates a significance of P < 0.01.

Article Snippet: According to the National Center for Biotechnology Information (NCBI) S. mutans genome annotation, the function of the majority of the differentially expressed genes (DEGs) are unknown.

Techniques:

Journal: Frontiers in Microbiology

Article Title: A GntR Family Transcription Factor in Streptococcus mutans Regulates Biofilm Formation and Expression of Multiple Sugar Transporter Genes

doi: 10.3389/fmicb.2018.03224

Figure Lengend Snippet: Scanning electron microscopy analysis reveals altered biofilm morphology and decreased biofilm extracellular matrix of S. mutans Δ stsR . Biofilms formed by S. mutans UA159, S. mutans Δ stsR , and complement strain were grown for 6 h and then scanned by scanning electron microscopy (SEM) under (A) 1000× magnification, (B) 5000× magnification, and (C) 20000× magnification.

Article Snippet: According to the National Center for Biotechnology Information (NCBI) S. mutans genome annotation, the function of the majority of the differentially expressed genes (DEGs) are unknown.

Techniques: Electron Microscopy

Journal: Frontiers in Microbiology

Article Title: A GntR Family Transcription Factor in Streptococcus mutans Regulates Biofilm Formation and Expression of Multiple Sugar Transporter Genes

doi: 10.3389/fmicb.2018.03224

Figure Lengend Snippet: Biofilm structure and EPS distribution of S. mutans strains observed by confocal microscopy. (A) Double-labeling of 6 h S. mutans biofilms. Green indicates bacteria (SYTO 9), and red indicates EPS (Alexa Fluor 647). Images were taken at 60× magnification. The three-dimensional reconstruction of the biofilms and the quantification of EPS/bacteria biomass were performed with IMARIS 7.0.0. (B) The ratio of EPS to bacteria at different heights was quantified with COMSTAT. Results are the average of five randomly selected positions of each sample and are presented as mean ± standard deviation. (C–E) Quantification of S. mutans UA159 (C) , S. mutans Δ stsR (D) , and Δ stsR/pDL278-stsR (E) biofilms. EPS biomass was performed with COMSTAT at different heights. Results are the average of five randomly selected positions of each sample and are presented as mean ± standard deviation.

Article Snippet: According to the National Center for Biotechnology Information (NCBI) S. mutans genome annotation, the function of the majority of the differentially expressed genes (DEGs) are unknown.

Techniques: Confocal Microscopy, Labeling, Bacteria, Standard Deviation

Journal: Frontiers in Microbiology

Article Title: A GntR Family Transcription Factor in Streptococcus mutans Regulates Biofilm Formation and Expression of Multiple Sugar Transporter Genes

doi: 10.3389/fmicb.2018.03224

Figure Lengend Snippet: Quantitative RT-PCR assays for the relative expression levels of gtfs and ftf genes in S. mutans UA159, S. mutans Δ stsR , and Δ stsR/pDL278-stsR . The experiments were carried out as described in Experimental procedures. All target genes were amplified using specific primers. Different gene expressions were normalized to the levels of 16S rRNA gene transcripts, and the folds of expression change were calculated.

Article Snippet: According to the National Center for Biotechnology Information (NCBI) S. mutans genome annotation, the function of the majority of the differentially expressed genes (DEGs) are unknown.

Techniques: Quantitative RT-PCR, Expressing, Amplification

Journal: Frontiers in Microbiology

Article Title: A GntR Family Transcription Factor in Streptococcus mutans Regulates Biofilm Formation and Expression of Multiple Sugar Transporter Genes

doi: 10.3389/fmicb.2018.03224

Figure Lengend Snippet: The differentially expressed sugar transport system operons in S. mutans Δ stsR . The genetic organization of differentially expressed gene clusters that were associated with sugar transport systems in S. mutans Δ stsR . Upregulated genes are colored red, and downregulated genes are colored green.

Article Snippet: According to the National Center for Biotechnology Information (NCBI) S. mutans genome annotation, the function of the majority of the differentially expressed genes (DEGs) are unknown.

Techniques:

Journal: Journal of Oral Biology and Craniofacial Research

Article Title: Effect of low-level diode laser on streptococcus mutans and lactobacillus acidophilus growth: An invitro study

doi: 10.1016/j.jobcr.2022.05.001

Figure Lengend Snippet: The interference effect of energy and time on S. mutans immediately after irradiation. The x-axis is the average number of colonies.

Article Snippet: Microorganisms Standard strains of S. mutans (PTCC:1683) and L. acidophilus (PTCC:1643) were purchased in the form of lyophilized vials from the collection center of fungi and bacteria of Iran Pasteur Institute.

Techniques: Irradiation

Journal: Journal of Oral Biology and Craniofacial Research

Article Title: Effect of low-level diode laser on streptococcus mutans and lactobacillus acidophilus growth: An invitro study

doi: 10.1016/j.jobcr.2022.05.001

Figure Lengend Snippet: The interference effect of energy and time on S. mutans 24 h after irradiation. The x-axis is the average number of colonies.

Article Snippet: Microorganisms Standard strains of S. mutans (PTCC:1683) and L. acidophilus (PTCC:1643) were purchased in the form of lyophilized vials from the collection center of fungi and bacteria of Iran Pasteur Institute.

Techniques: Irradiation