Junko Matsuyama

Nested PCR for detection of mutans streptococci in dental plaque

Aims: Mutans streptococci such as Streptococcus mutans and Streptococcus sobrinus have been implicated in human dental caries. In an attempt to develop a rapid and sensitive method for detecting Strep. mutans and Strep. sobrinus in dental plaque, a nested PCR amplification based on the 16S rRNA gene was employed.

PCR-restriction fragment length polymorphism analysis of genes coding for 16S rRNA in Veillonella spp.

Restriction fragment length polymorphism analysis of PCR-amplified 16S ribosomal DNA (16S rDNA PCR-RFLP) was used to generate restriction profiles of the American Type Culture Collection type strains of the genus Veillonella, i.e., V. atypica, V. caviae, V. criceti, V. dispar, V. parvula, V. ratti, and V. rodentium. Whole-cell protein profiles were obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis for comparative purposes. The 16S rRNA genes were amplified by PCR, and RFLP analysis of the 16S rDNA was performed with MnlI and Sau3AI. MnlI produced six RFLP patterns for seven type strains, since the patterns for V. atypica and V. caviae were the same. RFLP patterns with Sau3AI could distinguish between V. atypica and V. caviae. The type strains of Veillonella species were easily distinguished by 16S rDNA PCR-RFLP.

Differentiation of oral Actinomyces species by 16S ribosomal DNA polymerase chain reaction-restriction fragment length polymorphism

16S rDNA polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to generate restriction profiles of the reference strains, including the American Type Culture Collection type strains, of oral Actinomyces spp., i.e., A. israelii, A. gerencseriae, A. naeslundii genospecies 1 and 2, A. odontolyticus, A. meyeri and A. georgiae, and 23 Actinomyces strains isolated from human dental plaque. The 16S rRNA gene sequences from isolated genomic DNA samples were amplified by PCR. The PCR products were purified and characterized by single digestion with four restriction endonucleases, i.e., MnlI, HaeIII, CfoI, or HpaII. Among them, MnlI was found to discriminate the respective reference strains. The clinical isolates were assigned to one of the species, i.e., A. gerencseriae, A. naeslundii genospecies 1 and 2 and A. odontolyticus, on the basis of their restriction profiles by single digestion with MnlI. Thus, 16S rDNA PCR-RFLP, using MnlI, is a rapid and reliable method for the differentiation of oral Actinomyces spp.

Changes in the mouthful weights of familiar foods with age of five years, eight years and adults

Abstract For effective mastication and swallowing, to take optimum mouthful food regularly is necessary. The purpose of this study is to compare the weights and the variance of a mouthful food between children and adults. We studied mouthful weight and the coefficient variation in 5-year-old children (n=10), 8-year-old children (n=10) and adults (n=10) while they were eating rice, bread, sausage and apple on two different days. The test foods were served in random order 2–4 hours after lunch. Each portion was weighed before and after each bite to measure the mouthful weight, and the weight of the last bite of each portion was eliminated from the data. The mean weight and coefficient variation in each subject were calculated. The results showed the mean weight was largest in the adults, intermediate in the 8-year-old children, and smallest in the 5-year-old children for all test foods (ANOVA). Moreover, the mean coefficient variation among the weights of the groups revealed that mouthful weight within an individual varied most widely in the 5-year-old children and most narrowly in the adults, and that 8-year-old children could show the same coefficient variation of mouthful weight as adults in rice and apple (Steel-Dwass test). Our results suggest that mouthful weight becomes larger and more regularly with age.

Cultivable Anaerobic Microbiota of Infected Root Canals

Objective. Periapical periodontitis is an infectious and inflammatory disease of the periapical tissues caused by oral bacteria invading the root canal. In the present study, profiling of the microbiota in infected root canals was performed using anaerobic culture and molecular biological techniques for bacterial identification. Methods. Informed consent was obtained from all subjects (age ranges, 34–71 years). Nine infected root canals with periapical lesions from 7 subjects were included. Samples from infected root canals were collected, followed by anaerobic culture on CDC blood agar plates. After 7 days, colony forming units (CFU) were counted and isolated bacteria were identified by 16S rRNA gene sequencing. Results. The mean bacterial count (CFU) in root canals was (0.5 ± 1.1) × 106 (range 8.0 × 101–3.1 × 106), and anaerobic bacteria were predominant (89.8%). The predominant isolates were Olsenella (25.4%), Mogibacterium (17.7%), Pseudoramibacter (17.7%), Propionibacterium (11.9%) and Parvimonas (5.9%). Conclusion. The combination of anaerobic culture and molecular biological techniques makes it possible to analyze rapidly the microbiota in infected root canals. The overwhelming majority of the isolates from infected root canals were found to be anaerobic bacteria, suggesting that the environment in root canals is anaerobic and therefore support the growth of anaerobes.

Fermentation of Five Sucrose Isomers by Human Dental Plaque Bacteria

Sucrose has five structural isomers: palatinose, trehalulose, turanose, maltulose and leucrose. Although these isomers have been reported to be noncariogenic disaccharides, which cannot be utilized by mutans streptococci, there is no information about their fermentability by other bacteria in dental plaque. The purpose of the present study was to examine whether these isomers were fermented by predominant bacteria in human dental plaque. Clinical bacterial isolates obtained from dental plaque from 3 children aged 22 months to 50 months (146 strains) were inoculated into 3 ml of peptone-yeast extract (PY medium) containing glucose for 1 day, then an aliquot of 20 µl of culture medium was inoculated into 1 ml of PY medium containing 1% (w/v) of the respective test carbohydrates. After incubation for 1 day, the pH values and the optical density at 660 nm of the cultures were measured. Fermentation ability was measured by pH ≤5.5, growth by an OD660 of ≧0.5. Of the clinical isolates, 33% fermented palatinose, and 69% of these were Actinomyces species. All of the palatinose-fermenting bacterial strains fermented trehalulose, 25% fermented turanose, 70% fermented maltulose and 23% fermented leucrose. We therefore conclude that, in human dental plaque, there are significant numbers of bacteria that are able to ferment sucrose isomers.

Severe Impaction of the Primary Mandibular Second Molar Accompanied by Displacement of the Permanent Second Premolar

Tooth impaction is defined as any tooth that fails to erupt into a normal functional position and remains unerupted beyond the time at which it should normally erupt. Reports of impaction and eruption failure in primary teeth are relatively rare compared to permanent teeth. We report 2 rare cases where the second premolar was located on the occlusal side of the impacted mandibular second primary molar. In the first case, the succedaneous permanent tooth erupted after extraction of the primary tooth, fenestration, and traction. In the second case, the succedaneous permanent tooth erupted without fenestration or traction. Although the etiology of the tooth displacement was unknown in both cases, inhibition of the eruptive movement of the primary molar may have been associated with displacement of the succedaneous permanent premolar.

Cross-sectional analysis of age-related changes in the fluctuation of bite size

Abstract It has been reported that individual adults generally take consistently sized bites when eating the same food, while children do not. The present cross-sectional analysis was performed on 60 children and 20 adults to ascertain the age-related changes in the fluctuation in bite size and in the number of chews per bite. The subjects comprised four age groups (5-, 8-, and 11-year-old children and adults), with each group consisting of 10 males and 10 females. The subjects were instructed to take a bite of each of four test foods (bread, sausage, apple and rice), and they were allowed to chew and swallow as usual. After each bite, the remaining food was weighed to calculate the bite size. The fluctuations in bite size and in the number of chews were analyzed using coefficients of variation. The fluctuations in both bite size and the number of chews decreased with age, so that the 11-year-olds showed almost the same values as adults. The present results suggested that the physiological functions related to recognizing and consuming foods mature during early adolescence, when almost all permanent teeth have erupted. The present findings also indicated that the fluctuation in bite size might be an indicator of the maturation of masticatory function in children.

Hypophosphatasia: Evaluation of Size and Mineral Density of Exfoliated Teeth

OBJECTIVE Most cases of hypophosphatasia (HPP) exhibit early loss of primary teeth. Results of micro-computed tomography (micro-CT) analysis of teeth with HPP have not yet been reported. The purpose of the present study was to describe the size and mineral density distribution and mapping of exfoliated teeth with HPP using micro CT. STUDY DESIGN Seven exfoliated teeth were obtained from a patient with HPP. Exfoliated teeth sizes were measured on micro CT images and mineral densities of the mandibular primary central incisors were determined. RESULTS Partial dentures were fabricated for the patient to replace the eight primary teeth which had exfoliated. Most primary teeth sizes were within the normal range. The mean values of enamel and dentin mineral densities in teeth with HPP were 1.35 and 0.88 g/cm3, respectively, in the mandibular primary central incisors. CONCLUSION Mineral density distribution and mapping revealed that the values in teeth with HPP were lower than the homonymous teeth controls in all regions from the crown to apex. Furthermore, it was demonstrated that the differences between HPP and controls were larger on the crown side and the differences tended to converge on the apex side. These results suggested that the present patient showed mild hypomineralization in the primary dentition.

Oral Microbiota in Crevices Around Dental Implants: Profiling of Oral Biofilm

Large numbers of bacteria (>106/mm2) generally inhabit the surface of the oral cavity, particularly at the interface between teeth and gingiva, as an oral biofilm (microbiota). The establishment of anaerobic bacterial culture and molecular biological techniques has enabled us to isolate and detect various bacterial species from oral biofilm. It has been estimated that more than 600 bacterial species inhabit the oral cavity. Nevertheless, the oral cavity is considered healthy when the oral microbiota is composed of indigenous bacteria. Numerous environmental changes in the oral cavity may lead to accumulation of dental caries-associated or periodontitis-associated bacteria, resulting in the initiation of dental caries or periodontitis, respectively. The environment in crevices around dental implants is considered similar to that in subgingival sulcus, such as neutral pH, anaerobiosis and rich nutrition (e.g., amino acids and peptides). The environment may be supportive of anaerobic growth of the bacteria in microbiota in crevices around implants, particularly at the interface between histocompatible artificial material and mucosal epithelium. The microbiota may trigger inflammation in the tissue around the implants. In this article, the current topics on the profiling of oral microbiota in crevices around implants are reviewed.