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PubMed1986

Effects of chlorhexidine gluconate in drinking water on dental caries and oral microorganisms in the Syrian hamster.

Hamsters infected with Streptococcus sobrinus (formerly Streptococcus mutans) strain 6715-1119 were given high-sucrose diet 2000. After 5 days, they were presented with 0.05 or 0.1 per cent w/v chlorhexidine gluconate (CHX) continuously in their drinking water, which was well-tolerated. Compared to infected but untreated controls, their caries scores were less by 84 and 97 per cent respectively after 42 days. Coronal dental plaque deposits were reduced in a dose-related fashion. Culture plates of oral swabs showed reductions in Strep. sobrinus and total streptococci after 10 and 35 days of CHX. Hamster oral streptococci were more sensitive to CHX than were lactobacilli but neither was completely eliminated. The incidence of stained tooth deposits after 0.1 per cent CHX was slightly but not significantly greater than in controls.

Animals↗

PubMed2003

Quantitative evaluation of bacteria adherent to polyelectrolyte HEMA-based hydrogels.

The use of adhesive poly(HEMA)-based hydrogels is standard practice in dental restorative procedures. Microorganisms, which potentially can cause oral pathologies, may colonize these polymers. In the present work, bacterial adhesion to polymers prepared with 2-hydroxyethyl methacrylate (HEMA) and to different molar ratios of 2-acrylamido-2-methylpropane-sulfonic acid (AMPS) and/or to 2-methacryloyloxyethyl-tri-methyl-ammonium chloride (METAC) co-monomers were tested. A colorimetric assay system that utilizes the Microbo revelation medium (Microbo srl, Rome, Italy) for microbial counts is shown to be capable of counting the number of adherent bacterial cells without removing them from polymer surfaces. In conditions that mimic those present in the oral cavity, similar bacterial adhesion percentages on the same polymer were observed with the different bacteria belonging to both gram-positive and gram-negative genera, such as Streptococcus sobrinus and Streptococcus oralis (resident microorganisms in the oral cavity) and Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa (transient microorganisms in the oral cavity). It is determined that the physico-chemical characteristics of poly(HEMA)-based hydrogels are the major factors promoting bacterial adhesion, which increased with increasing water content in the swollen polymers, reaching maximal values on the cationic polymers.

Bacteria↗

PubMed1994

Action of sodium fluoride on phagocytosis by systemic polymorphonuclear leucocytes.

Alterations in phagocytosis appear to be important in the onset and development of periodontitis. We investigated new substances that may be of use in the treatment of periodontitis. In a preliminary study, we tested the effect of sodium fluoride on phagocytosis by circulating polymorphonuclear leucocytes (PMNs) in 10 replicate assays using blood from six healthy subjects. Sodium fluoride was tested at concentrations of 1.0 micron to 4.0 microns against Streptococcus oralis, Streptococcus mutans, Streptococcus sobrinus and Streptococcus sanguis. The phagocytic index against all microorganisms increased significantly at all concentrations of sodium fluoride assayed; this effect was dose dependent. Sodium fluoride appeared to stimulate phagocytosis via two mechanisms: an apparent increase in bacterial susceptibility to phagocytosis, and direct stimulation of phagocytosis by PMNs.

Adult↗

PubMed2003

Kuwanon G: an antibacterial agent from the root bark of Morus alba against oral pathogens.

Kuwanon G was isolated from the ethyl acetate fraction of methanol extract of Morus alba and its structure was elucidated by 13C-NMR, 1H-NMR and FAB-MS. Antibacterial activity of kuwanon G was investigated by the minimum inhibitory concentration (MIC) test and the viable cell count method. MIC of kuwanon G against Streptococcus mutans causing dental caries was determined to be 8.0 microg/ml. The bactericidal test showed that kuwanon G completely inactivated S. mutans at the concentration 20 microg/ml in 1 min. Kuwanon G also significantly inhibited the growth of other cariogenic bacteria such as Streptococcus sobrinus and Streptococcus sanguis, and Porpyromonas gingivalis causing periodontitis. Transmission electron microscopy (TEM) of kuwanon G treated cells demonstrated remarkable morphological damage of the cell wall and condensation of the cytoplasm.

Anti-Bacterial Agents↗

PubMed2003

Classification of oral streptococci by two-dimensional gel electrophoresis with direct activity stain for glycosyltransferases.

Thirty eight strains of oral streptococci were divided into six types by two-dimensional gel electrophoresis (2-DE) followed by glycosyltransferase (GLT) activity stain: Type 1, Streptococcus mutans; Type 2, Streptococcus rattus; Type 3, Streptococcus sobrinus and Streptococcus downei; Type 4, Streptococcus cricetus; Type 5, Streptococcus salivarius; and Type 6, Streptococcus sanguis, Streptococcus oralis and Streptococcus gordonii. In Types 1, 2 and 5, two glucosyltransferases synthesizing water-insoluble (GTF-I) and water-soluble glucans (GTF-S) and a fructosyltransferase (FTF) were detected. In Types 3 and 4, GTF-I and two GTF-Ss were detected. Only one GTF-S was detected in Type 6. The 2-DE patterns for these six types were clearly distinguished from each other based on the kind, number and location of GLTs in gel.

Bacterial Typing Techniques↗

PubMed2005

Susceptibilities of periodontopathogenic and cariogenic bacteria to antibacterial peptides, {beta}-defensins and LL37, produced by human epithelial cells.

OBJECTIVES: Antimicrobial peptides are one of the factors involved in innate immunity. The susceptibility of periodontopathogenic and cariogenic bacteria to the major antimicrobial peptides produced by epithelia was investigated. METHODS: Synthetic antimicrobial peptides of human beta-defensin-1 (hBD1), hBD2, hBD3 and LL37 (CAP18) were evaluated for their antimicrobial activity against oral bacteria. They included Actinobacillus actinomycetemcomitans (20 strains), Porphyromonas gingivalis (6), Prevotella intermedia (7), Fusobacterium nucleatum (7), Streptococcus mutans (5), Streptococcus sobrinus (5), Streptococcus salivarius (5), Streptococcus sanguis (4), Streptococcus mitis (2) and Lactobacillus casei (1). RESULTS: Although the four peptides had bactericidal activity against all bacteria tested, the degree of antibacterial activity was variable against the different strains and species. The antibacterial activity of hBD1 was lower than that of the other peptides. Among the bacteria tested in this study, F. nucleatum was highly susceptible to hBD3 and LL37, and S. mutans was highly susceptible to hBD3. We measured the Zeta-potential, representing the net charge of whole bacteria, to study the relationship between susceptibility to cationic peptide and the net charge of the bacteria. Although we found some correlation in A. actinomycetemcomitans strains, we did not find a definite correlation with all the bacterial species. CONCLUSIONS: These results indicate that beta-defensins and LL37 have versatile antibacterial activity against oral bacteria.

Amino Acid Sequence↗

PubMed2004

Molecular characterization of dextranase from Streptococcus rattus.

The complete nucleotide sequence of the dextranase gene of Streptococcus rattus ATCC19645 was determined. An open reading frame of the dextranase gene was 2,760 bp long and encoded a dextranase protein consisting of 920 amino acids with a molecular weight of 100,163 Da and an isoelectric point of 4.67. The S. rattus dextranase purified from recombinant Escherichia coli cells showed dextran-hydrolyzing activity with optimal pH (5.0) and temperature (40 C) similar to those of dextranases from Streptococcus mutans and Streptococcus sobrinus. The deduced amino acid sequence of the S. rattus dextranase revealed that the dextranase molecule consists of two variable regions and a conserved region. The variable regions contained an N-terminal signal peptide and a C-terminal cell wall sorting signal; the conserved region contained two functional domains, catalytic and dextran-binding sites. This structural feature of the S. rattus dextranase is quite similar to that of other cariogenic species such as S. mutans, S. sobrinus, and Streptococcus downei.

Amino Acid Sequence↗

PubMed1989

Molecular characterization of a surface protein antigen gene from serotype c Streptococcus mutans, implicated in dental caries.

The complete nucleotide sequence of the gene for a cell-surface protein antigen (PAc) of Streptococcus mutans MT8148 (serotype c) was determined. The pac gene consisted of 4695 bp and coded for a 170773D protein. The pac gene product contained a putative 38 amino acid signal peptide, resulting in a 166817D mature protein. A potential promoter sequence and a putative Shine-Dalgarno sequence preceded the open reading frame. Two internal repeating amino acid sequences were present in the PAc. One repeating region located in the N-terminal region was rich in alanine, and the other located in the central region was rich in proline. Southern blot analysis under the less stringent condition (allowing up to 35% base mismatch) revealed that the probe covering the proline-rich region hybridized to DNA preparations from strains of Streptococcus cricetus, Streptococcus sobrinus and Streptococcus downei as well as Streptococcus mutans.

Amino Acid Sequence↗

PubMed1990

A IIIman protein is involved in the transport of glucose, mannose and fructose by oral streptococci.

We show in this article that the transport of glucose, mannose and fructose by the phosphoenolpyruvate: mannose phosphotransferase system of oral streptococci requires the participation of a protein component that we have called IIIman. This protein was purified from Streptococcus salivarius by chromatography on DEAE-cellulose, DEAE-TSK, hydroxyapatite, and Dyematrex Green A. The purified protein migrated as a 38,900 molecular weight protein on a sodium dodecyl sulfate polyacrylamide gel. However, electrophoretic analysis of phosphoproteins and Western blot experiments indicated the presence in membrane-free cellular extracts of S. salivarius of 2 different forms of IIIman having molecular weights of 38,900 and 35,200. The presence of the high-molecular-weight form of IIIman was observed by immunodiffusion, Western blot and phosphorylation by [32]PEP in S. salivarius, Streptococcus mutans, Streptococcus sobrinus, and Streptococcus lactis but not in Streptococcus faecium, Staphylococcus aureus, Bacillus subtilis and Lactobacillus casei. Antibodies directed against the IIIman of S. salivarius did not react with the IIIman of Escherichia coli.

Bacterial Proteins↗

PubMed1998

Acid-regulated proteins induced by Streptococcus mutans and other oral bacteria during acid shock.

Our previous research has demonstrated that with the more aciduric oral bacteria, an acid shock to sub-lethal pH values results in the induction of an acid tolerance response that protects the cells at extremely low pH (pH 3.0-4.0) that kills unadapted control cells maintained at pH 7.5 (Oral Microbiol Immunol 1997: 12: 266-273). In this study, we were interested in comparing the protein profiles of acid-shocked and control cells of nine organisms from three acid-ogenic genera that could be categorized as strong, weak and non-acid responders in an attempt to identify proteins that could be classified as acid-regulated proteins and which may be important in the process of survival at very low pH. For this, log-phase cultures were rapidly acidified from pH 7.5 to 5.5 in the presence of [14C]-amino acids for varying periods up to 2 h, the period previously shown to be required for maximum induction of the acid response. The cells were extracted for total protein and subjected to one-dimensional sodium dodecyl sulfate-polyacrylamide chromatography with comparable control and acid-shocked protein profiles compared by scanning and computer analysis. Of particular interest were the proteins in the acid-shocked cells that showed enhanced labeling (i.e., synthesis) over the control cells, since these were considered acid-regulated proteins of importance in pH homeostasis. Streptococcus mutans LT11 generated the most rapid and complex pattern: a total of 36 acid-regulated proteins showing enhanced synthesis, with 25 appearing within the first 30 min of acid shock. The enhanced synthesis was transient with all proteins, with the exception of two with molecular weights of 50/49 and 33/32 kDa. Within the acid-regulated proteins were proteins having molecular weights comparable to the heat shock proteins and the various subunits of the membrane H+/ATPase. By comparison, the strong responder, Lactobacillus casei 151, showed the enhanced formation of only nine proteins within the first 30 min of the acid shock, with a total of 11 acid-regulated proteins formed during the 2-h adaptation period with enhanced synthesis transient for seven of these proteins. Streptococcus salivarius AT2 and Streptococcus gordonii TH12 had the formation of 6 and 8 proteins enhanced, while the weakly responding organisms, Streptococcus sanguis ATCC 10,556 and Streptococcus oralis ATCC 10,557, exhibited 8 and 6 such proteins, respectively. Even non-responding strains unable to survive at very low pH, such as Streptococcus sobrinus CH125/43, Streptococcus mitis ATCC 12,261 and Actinomyces naeslundii 301-13 showed the initial formation of 3-9 acid-regulated proteins, but protein synthesis was not sustained over the entire adaptation period. Clearly, the survival of oral bacteria at very low pH is related, not to the total number of the acid-regulated proteins induced per se but to the formation of key proteins that function to augment normal pH homeostasis.

Acids↗

PubMed2006

Human T-cell responses to oral streptococci in human PBMC-NOD/SCID mice.

We investigated cellular and humoral immune responses to oral biofilm bacteria, including Streptococcus mutans, Streptococcus anginosus, Streptococcus sobrinus, and Streptococcus sanguinis, in NOD/SCID mice immunized with human peripheral blood mononuclear cells (hu-PBMC-NOD/SCID mice) to explore the pathogenicity of each of those organisms in dental and oral inflammatory diseases. hu-PBMC-NOD/SCID mice were immunized by intraperitoneal injections with the whole cells of the streptococci once a week for 3 weeks. FACS analyses were used to determine the percentages of various hu-T cell types, as well as intracellular cytokine production of interleukin-4 and interferon-gamma. Serum IgG and IgM antibody levels in response to the streptococci were also determined by enzyme-linked immunosorbent assay. S. anginosus induced a significant amount of the proinflammatory cytokine interferon-gamma in CD4(+) and CD8(+) T cells in comparison with the other streptococci. However, there was no significant differences between the streptococci in interleukin-4 production by CD4(+) and CD8(+) T cells after inoculation. Further, S. mutans significantly induced human anti-S. mutans IgG, IgG(1), IgG(2), and IgM antibodies in comparison with the other organisms. In conclusion, S. anginosus up-regulated Th1 and Tc1 cells, and S. mutans led to increasing levels of their antibodies, which was associated with the induction of Th2 cells. These results may contribute to a better understanding of human lymphocyte interactions to biofilm bacteria, along with their impact on dental and mucosal inflammatory diseases, as well as endocarditis.

Animals↗

PubMed1983

Prevalence, distribution of serotypes, and cariogenic potential in hamsters of mutans streptococci from elderly individuals.

The prevalence of mutans streptococci (Streptococcus mutans, Streptococcus cricetus, Streptococcus sobrinus, and Streptococcus rattus) was determined in the salivas of 169 elderly individuals ranging in age from 60 to 87 years. Approximately 40% of these individuals were edentulous and wore full upper and lower dentures. With the exception of a higher proportion of saliva counts below 1,000 CFU/ml in the full-denture wearers, the prevalence and the serotype and species distributions of the mutans streptococci were similar in the denture wearers and individuals with natural teeth only. The species and serotype distributions of mutans streptococci in this elderly population were also consistent with reported observations of other workers on younger, more caries-prone populations. A total of 87 representative isolates of the mutans streptococci were tested for cariogenic potential in a hamster model system. A considerable degree of variation in virulence between different strains was observed. However, these differences were not relatable to individual species or serotypes or to whether the organisms were isolated from denture wearers or naturally dentate subjects. The results of our studies indicate that elderly individuals with either natural or artificial dentitions may be a hitherto unrecognized reservoir of mutans streptococci having varying degrees of potential cariogenicity. Hence, in close family situations they could serve, along with parents and siblings, as vectors in the initial transmission of cariogenic microorganisms to young children.

Aged↗

PubMed1988

Effect of nutritional constraints on the biosynthesis of the components of the phosphoenolpyruvate: sugar phosphotransferase system in a fresh isolate of Streptococcus mutans.

A procedure for the purification of enzyme I (EI) and the protein HPr, the general components of the phosphoenolpyruvate:sugar phosphotransferase system, from Streptococcus mutans serotype c is presented. The method was also applied successfully to the purification of EI and HPr from Streptococcus salivarius, Streptococcus sobrinus, and Streptococcus sanguis. Using specific antibodies obtained against the proteins purified from S. mutans DR0001, we determined quantitatively by rocket electrophoresis the cellular levels of EI and HPr in a freshly isolated strain of S. mutans grown under various conditions in continuous culture. The activity of a few specific EIIs was also determined by an in vitro phosphorylation test. Results indicated that maximum EII activities for glucose, mannose, and 2-deoxyglucose were obtained under conditions of glucose limitation, at pH 7.0 and low dilution rate (D = 0.057/h). Increasing the amount of glucose or the dilution rate (D = 0.40/h) or decreasing the pH from 7.0 to 5.5 resulted in a 1.4- to 24-fold decrease in these activities. The EII activity for fructose was not influenced by the growth conditions in the same way as the other EIIs. The fructose EII was highest at pH 5.5 and at high dilution rate under conditions of glucose or nitrogen limitation and was always repressed at pH 7.0 and at low dilution rates. The intracellular levels of EI were also dependent on the growth conditions. The highest concentration (0.65 nmol/mg of protein) was observed in cells grown under glucose limitation at pH 7.0 and high dilution rate, and the lowest concentration (0.12 nmol/mg of protein) was found in cells grown under glucose excess at pH 7.0 and high dilution rate. The other general component of the phosphoenolpyruvate:sugar phosphotransferase system, the protein HPr, was not influenced significantly by varying growth conditions.

Bacterial Proteins↗

PubMed1989

[Proliferation and superoxide dismutase (SOD) induction of Streptococcus mutans (sobrinus) B13. 1. Metal requirement].

Many of micro-organisms under aerobic conditions showed oxygen tolerance caused by the induction of several oxygen-derived free radical scavengers. In the present study, we tried to clarify the changes of active oxygen metabolism of streptococcus mutans (sobrinus) B13 under aerobic conditions. Aerobic and anaerobic culture of streptococcus mutans (sobrinus) B13 was made in normal, metal chleted, and metal supplemented medium. Superoxide dismutase (SOD) activity and bacterial proliferation were determined on the cultured cells. We obtained the following results. 1) In the anaerobic normal medium, SOD activity was not detected, but proliferation was observed. On the other hand, the neither were detected in the metal chleted medium. 2) No effect was observed on the induction of SOD activity and proliferation by the supplementation of copper in the metal chelated medium. 3) We observed positive effects on the induction of SOD activity and proliferation by the supplementation of iron, managnese, or both in the metal chlated medium under aerobic condition. Based on the above results, we found that iron and manganese were essential metals for SOD activity induction. Moreover, we suggested that there was close association of the aerobic bacterial proliferation with the induction of oxygen-derived free radical scavengers.

Energy Metabolism↗

PubMed1998

Concentrated bovine colostral whey proteins from Streptococcus mutans/Strep. sobrinus immunized cows inhibit the adherence of Strep. mutans and promote the aggregation of mutans streptococci.

The aim of this study was to examine the effect of bovine colostral whey proteins from cows immunized with Streptococcus mutans/Strep. sobrinus on the adherence and aggregation of caries-inducing bacteria, i.e., mutants streptococci. Both adherence and aggregation are important phenomena in the bacterial colonization of the human oral cavity. In all adherence experiments there was a significant difference between treatments by immune product (IP; from immunized cows) and a control product (CP; a similar product from non-immunized cows). The adherence of 35S-labelled Strep. mutans cells (serotype c) to parotid saliva-coated hydroxyapatite (SHA) was dose-dependently inhibited by both IP and CP if SHA was coated with either product before exposure to bacteria, but markedly lower concentrations of IP than CP were effective. When instead of SHA the bacterial cells were pretreated with IP or CP, only IP strongly and dose-dependently inhibited streptococcal adherence. When bacteria, IP or CP, and SHA were incubated simultaneously, a significant difference between IP and CP treatments was again found. Further, IP effectively aggregated both Strep. mutans and Strep. sobrinus cells, whereas hardly any effect was seen with CP. Both IP and CP aggregated the control bacterium Strep. sanguis, which affected the adherence of the pretreated bacteria.

Animals↗

PubMed1995

Sucrose retention and colonization by mutans streptococci at different sites of the dentition.

The correlations between the retention and elimination of sucrose at different oral sites and the colonization by mutans streptococci at the same sites were tested in 10 subjects with > 10(5) mutans streptococci per millilitre of saliva. Paraffin-stimulated saliva was collected and plaque samples were taken with a toothpick from all the buccal surfaces except the third molars. One week after the plaque sampling, the sucrose clearance was determined. After a mouth rinse with 10 ml of a 10% sucrose solution, pre-weighed filter paper discs were placed at representative buccal sites of the interdental papilla between molars, premolars and incisors in the upper and lower jaw. The samples were collected 0.5, 2.5, 5.5, 8.5 and 12.5 min after rinsing. Analysis of the sucrose concentration was made using an enzymatic method. The oral sugar clearance was slower in the anterior region of the upper jaw than in the posterior regions and in the central region of the lower jaw. The frequency of mutans streptococci decreased towards the anterior teeth, with Streptococcus sobrinus predominating over Streptococcus mutans. A negative correlation was observed in the upper jaw between sugar clearance and retention and the prevalence of mutans streptococci, whereas the opposite tendency was observed in the lower jaw.

Adult↗

PubMed1999

Effect of salivary biofilm on the adherence of oral bacteria to bleached and non-bleached restorative material.

OBJECTIVE: The objective of this work was to examine the effect of in vitro salivary biofilm on the adherence of oral bacteria to bleached and non-bleached restorative material (Charisma). METHODS: Charisma samples, prepared in silicon models, were treated with either 10% carbamide peroxide (CP) or 10% hydrogen peroxide (HP). After incubation with the bleaching agent for a period of one, two or three days, the samples were coated with freshly collected human saliva. The adsorption pattern of the saliva to the restorative material was determined using gel electrophoresis coupled with computerized densitometry techniques. The amount of salivary proteins adsorbed onto the treated surfaces was measured using the Bradford method. Sucrose-dependent bacterial adhesion to the salivary-coated Charisma was tested using radio-labeled Streptococcus mutans, Streptococcus sobrinus and Actinomyces viscosus. Adhesion of each bacterium to surfaces pretreated with the bleaching agents was compared with saliva coated bleached surfaces. RESULTS: The profile of salivary proteins adsorption followed a similar pattern in Charisma samples pretreated with either CP or HP or untreated samples. However, the total amount of salivary proteins adsorbed onto the samples decreased after bleaching with CP or HP. Salivary biofilm, coating the surface of the restorative material, significantly decreased sucrose-dependent adhesion of Streptococcus sobrinus and Streptococcus mutans to the bleached and non-bleached surfaces, compared to non-coated specimens (p < 0.05). Saliva had a minor effect on adhesion of Actinomyces viscosus. SIGNIFICANCE: Our study demonstrates the importance of salivary biofilm in controlling adhesion of oral bacteria to restorative material pretreated with bleaching agents or untreated.

Actinomyces viscosus↗

PubMed1998

Bacterial adherence to bleached surfaces of composite resin in vitro.

OBJECTIVE: The effect of bleaching agents on bacterial adherence to polished surfaces of composite resin restorations was assessed in vitro. STUDY DESIGN: Samples of light-curing composite resins were treated with either 10% carbamide peroxide or 10% hydrogen peroxide for 1, 3, or 7 days. Bacterial adherence of Streptococcus mutans, Streptococcus sobrinus, and Actinomyces viscosus to the treated resin samples was analyzed and compared with adherence to nonbleached controls. RESULTS: A 10% solution of carbamide peroxide caused a significant increase in surface adherence of Streptococcus mutans and Streptococcus sobrinus after 3 days (P < .01). A 10% solution of hydrogen peroxide caused a significant increase in surface adherence of Streptococcus mutans and Streptococcus sobrinus after 3 and 7 days (P < .01). A decrease in adherence of Actinomyces viscosus was found after treatment with 10% hydrogen peroxide for 7 days (P < .05). CONCLUSIONS: It appears that bleaching agents may affect adherence of certain cariogenic microorganisms to the outer surfaces of composite resin restorations.

Actinomyces viscosus↗