Comparison of glucan-binding proteins in cariogenicity of Streptococcus mutans

Streptococcus mutans has been implicated as a primary causative agent of dental caries in humans. Bacterial components associated with the adhesion phase of S. mutans include cell-associated and cell-free glucosyltransferases (GTFs), as well as protein antigen c and proteins that bind glucan. At least four types of S. mutans glucan-binding protein (Gbp) have been identified; GbpA, GbpB, GbpC and GbpD. In the present study, GbpA-, GbpB- and GbpC-deficient mutants (AD1, BD1 and CD1, respectively) were constructed, and their cariogenic properties were evaluated by comparing them to those of their parent strain MT8148. All of the Gbp mutants showed lower levels of dextran binding, while the sucrose-dependent adhesion levels of AD1 and CD1 were lower than in the parental strain. The expression of each GTF was detected in the Gbp mutants, however, they had lower levels of cell-free-GTF activity than the parental strain. On the other hand, in acid tolerance assays, BD1 was the most sensitive among all of the tested strains. These results suggest that GbpA and GbpC in S. mutans have strong relationships with cariogenicity, while GbpB may have another biological function.     

Microscopic and crystallographic examinations of the teeth of the X-linked hypophosphatemic mouse

Teeth from the C57BL/6J-Hyp mouse (Hyp mouse) were examined histologically, radiographically, and crystallographically. Microscopic examinations of the ground sections, the decalcified and H-E-stained sections, and the contact microradiograms of molars and incisors from the Hyp mouse showed several abnormalities--such as large pulp chamber, wide predentin, thin dentin at the pulp floor, and multiple occurrences of interglobular dentin. Powder and microbeam x-ray diffraction analyses showed that the crystallinity of hydroxyapatite in incisor globular dentin from the Hyp mouse was higher than that in incisor dentin from the normal mouse. On the other hand, the findings of transmission electron microscopy demonstrated that the sizes of hydroxyapatite crystals of globular dentin in the Hyp incisor were larger than those in normal incisor dentin. These results demonstrated that the findings in Hyp mouse teeth were in accord with those of human XLH teeth.     

Efficacy of mandibular manipulation technique for temporomandibular disorders patients with mouth opening limitation: a randomized controlled trial for comparison with improved multimodal therapy

Purpose

Manual therapy has been used for the treatment of patients with temporomandibular disorders (TMD) with mouth-opening limitations. However, the curative effect of manipulation differs among researchers, and its necessity remains controversial. The purpose of this study was to confirm the efficacy of manipulation using a randomized controlled trial (RCT).

Crown dilaceration of a first premolar caused by extraction of its deciduous predecessor: a case report

This report describes a rare case of crown dilaceration of the mandibular first premolar caused by trauma during extraction of the precedent primary first molar. The mandibular first primary molar had been extracted at the age of 4 years 7 months. Compared to the pre-operative radiograph, the post-operative film showed that the direction of the first premolar tooth germ had changed, suggesting that crown dilaceration had been induced by the surgical procedures during tooth extraction. From radiographic examinations, the premolar was considered to have erupted spontaneously.     

Demonstration of mother-to-child transmission of Streptococcus mutans using multilocus sequence typing

A new reliable genotyping method, multilocus sequence typing (MLST), was used to evaluate vertical transmission of the cariogenic pathogen Streptococcus mutans. A total of 136 S. mutans strains were isolated from saliva samples of 20 Japanese mother-child pairs, including 5 girls and 5 boys with primary dentition, and 5 girls and 5 boys with mixed dentition. The nucleotide sequences of 8 partial housekeeping genes, aroE, murI, gltA, glnA, glk, tkt, lepC, and gyrA, were analyzed and a similarity for all of those sequences between strains from a mother-child pair was regarded as indicating transmission, which was shown in 70% of the pairs. Interestingly, the rate of transmitted strains from mothers was significantly higher in the girls (90%) than in the boys (p = 0.001). Furthermore, the S. mutans sequence type (ST) with the highest distribution percentage in each maternal saliva sample was found to be transferred to their children. In addition, variations in two large conjugative-transfer associated regions, TnSmu1 and TnSmu2, were determined and compared with the STs defined by MLST. No variations in those two regions shown by PCR patterns were present in any of the strains isolated from the same families with the same STs, though isolates of some STs from different families showed distinct patterns for TnSmu2. Our results indicate that mothers are the main source for transmission of S. mutans to their children, while the present MLST method was also shown to be useful for investigating bacterial transmission.     

Detection and serotype distribution of Actinobacillus actinomycetemcomitans in cardiovascular specimens from Japanese patients

Actinobacillus actinomycetemcomitans, an important pathogen in periodontitis, has also been detected in cardiovascular tissues. Sixty heart valves were collected during valve replacement surgery from 60 patients (one from each), 10 were from patients with infective endocarditis (IE group) and 50 were from patients with other valvular diseases (non-IE group). In addition, 46 samples of aneurysmal tissue were taken from 46 patients with a thoracic or abdominal aneurysm (Aneurysm group, one from each). Dental plaque samples were taken from 54 of the patients, 31 in the IE and non-IE groups and 23 in the aneurysm group. First, the distribution of A. actinomycetemcomitans in all specimens was analysed using a polymerase chain reaction method, which resulted in a positive reaction in 33 (31.1%) of the cardiovascular specimens and 25 (46.3%) of the dental plaque samples. Next, using serotype-specific sets of primers, the serotype distribution of A. actinomycetemcomitans in the cardiovascular specimens and dental plaque samples was found to be significantly different compared to dental plaque samples from Japanese subjects reported previously.     

Interaction of structural isomers of sucrose in the reaction between sucrose and glucosyltransferases from mutans streptococci

Structural isomers of sucrose, i.e. disaccharides composed of glucose and fructose molecules with different glucosidic linkages, were examined for their effect on the reaction between sucrose and various glucosyltransferases (GTases) from Streptococcus mutans MT8148 and Streptococcus sobrinus 6715. Trehalulose (alpha 1-1), turanose (alpha 1-3), maltulose (alpha 1-4), and palatinose (alpha 1-6) were used as the sucrose analogues. Mutans streptococci were found not to utilize these sucrose analogues. Analysis of enzymatic products of GTase and sucrose with thin layer chromatography clearly revealed that glucan synthesis from [14C]sucrose by the various purified GTase preparations from S. mutans and S. sobrinus was inhibited in the presence of these sucrose analogues except turanose, resulting in the release of increased amounts of [14C]fructose and [14C]oligosaccharides. It was also found that the fructose residues in the oligosaccharides were derived from those of sucrose analogues but not sucrose itself. The Lineweaver-Burk plots of the substrate saturation kinetics of GTase vs sucrose indicated increased Km and Vmax in the presence of sucrose analogue, as compared with sucrose alone. Finally, these sucrose analogues except turanose inhibited sucrose dependent cellular adherence of S. sobrinus 6715 to a glass surface, while they scarcely inhibited the adherence of S. mutans MT8148. Among the analogues, maltulose appeared the most effective inhibitor against GTases in general.     

Virulence factors of Streptococcus mutans and dental caries prevention

Streptococcus mutans possesses the abilities to adhere to pellicle-coated tooth surfaces and to form acids - two characteristics associated with the cariogenicity of this micro-organism. De novo synthesis of insoluble glucan by S. mutans glucosyltransferase from sucrose is essential in the adherence process. Therefore, agents which interfere with the adherence ability of S. mutans would be useful for controlling dental caries. In the present report, we have summarized our recent findings concerning virulence factors of S. mutans and means for prevention of S. mutans-induced dental caries.     

Effects of hyposalivation on the oral microflora of rats fed sucrose or wheat flour diets.

The effects of hyposalivation on the induction of dental caries and on the composition of the oral microflora were examined in specific pathogen-free Sprague-Dawley rats fed either a sucrose or a wheat flour diet with or without inoculation of Streptococcus mutans. Significant dental caries was induced in hyposalivated rats fed diet 2000 containing 56% sucrose, irrespective of infection by S. mutans. Diets containing 56% wheat flour did not induce dental caries in either hyposalivated or sham-operated rats, irrespective of infection by S. mutans. Bacteriological examinations at the end of the experiment demonstrated that the total numbers of lactobacilli and staphylococci increased in hyposalivated rats irrespective of the diet given, while the inoculated stain of S. mutans decreased significantly in hyposalivated rats. These findings suggest that some acidogenic microorganisms such as lactobacilli and staphylococci that can utilize sucrose or glucose but not wheat flour may also promote dental caries in hyposalivated rats.     

Mutacin activity of strains isolated from children with varying levels of mutants streptococci and caries

A total of 157 isolates of mutants streptococci from plaque and saliva of 94 children were studied for their serotypes, mutacin production, frequency and spectrum of activity. Of these isolates 71% were identified as serotype c and 22% as serotype e. Serotypes f, d and g, and one untypable strain made up about 7% of the isolates. More than one serotype was found in 13% of the children. Mutacin was produced by 83% of the isolates against one or more of the 14 indicator strains representing mutants streptococci. Streptococcus sanguis, Strep. oralis, Strep. gordonii, Strep. salivarius and Strep. pyogenes. Isolates that had a broad inhibitory spectrum also produced larger inhibition zones than isolates that inhibited fewer strains. When evaluating the effect of mutacin in vivo on plaque ecology, it was found that the counts of mutans streptococci or the proportion of mutans streptococci in the total streptococcal count of plaque did not differ between plaques containing strains that produced much mutacin and those with little production. The findings also failed to reveal an association between caries experience and mutacin activity.