Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

OBJECTIVES: The survival of bacteria in the sports drink and orange juice remaining in and at the mouth of bottles after direct drinking was examined after immediately drinking and incubation at 37°C for 24 h. METHODS: Nine healthy participants were asked to drink approximately 100 mL of a plastic bottled sports drink or orange juice. The samples were cultured anaerobically at 37°C for 7 days. Genomic DNA was extracted from the resulting individual colonies, and bacterial species were identified using 16S rRNA gene sequencing. RESULTS: The mean amount of bacteria in the remaining sports drink and orange juice, immediately after drinking, were (1.6 ± 2.3) × 103 colony-forming units (CFU)/mL and (2.9 ± 3.3) × 103 CFU/mL, respectively. Additionally, bacteria recovered from the mouths of the sports drink and orange juice bottles were (2.5 ± 5.5) × 104 CFU/mL and (5.8 ± 2.4) × 103 CFU/mL, respectively. Oral bacteria, such as Streptococcus, Actinomyces, Neisseria, and Rothia were found to be transferred in the sports drink and orange juice, and the bacteria were scarcely detected after incubation at 37°C for 24 h. CONCLUSIONS: The bacterial levels differed significantly from the previously reported levels in bottled tea 24 h after drinking, suggesting that remaining drinks with low pH levels can be preserved for a longer period.

DOI： 10.1016/j.job.2022.08.003

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OBJECTIVES: Breast milk is a valuable and useful source of nutrition; however, surplus milk is routinely discarded for hygiene reasons despite an unclear scientific basis. Here, we profiled the microbiota of expressed breast milk before and after feeding with an artificial nipple and examined the bacterial survival in breast milk stored at 4°C. METHODS: Eleven mother-baby pairs were included in the study. Samples of expressed breast milk were collected before and after feeding with an artificial nipple and examined both immediately (0 h) and after storage for 3 and 12 h at 4°C. Each sample was inoculated onto a blood agar plate and incubated anaerobically and aerobically at 37°C. Genomic DNA was extracted from individual bacterial colonies, which were identified by 16S rRNA gene sequencing. RESULTS: Before feeding, the bacterial counts at 0 and 12 h were (1.4 ± 1.6) × 105 colony-forming units (CFU)/mL and (1.4 ± 0.6) × 105 CFU/mL, respectively. Staphylococcus (47.7% and 41.9%, respectively), Cutibacterium (20.7% and 36.0%, respectively), and Streptococcus (16.1% and 6.6%, respectively) were identified among the samples. In contrast, after feeding, the bacterial counts at 0 and 12 h were (2.7 ± 1.7) × 105 CFU/mL and (2.1 ± 2.5) × 105 CFU/mL, respectively. Staphylococcus (30.1% and 37.4%, respectively), Cutibacterium (11.7% and 31.7%, respectively), and Streptococcus (41.5% and 25.2%, respectively), were identified among the samples. CONCLUSIONS: Bacteria were present in the breast milk before feeding. Although the main component of the microbiota shifted from Staphylococcus to Streptococcus species after feeding, these results suggest that surplus expressed breast milk may be preserved safely in a refrigerator for at least 12 h after feeding with an artificial nipple.

DOI： 10.1016/j.job.2022.09.004

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Authorship：Last author,　Corresponding author   Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.job.2022.07.002

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.job.2021.05.003

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：MDPI AG  

It has been speculated that oral bacteria can be transferred to tea in plastic bottles when it is drunk directly from the bottles, and that the bacteria can then multiply in the bottles. The transfer of oral bacteria to the mouth of bottles and bacterial survival in the remaining tea after drinking directly from bottles were examined immediately after drinking and after storage at 37 °C for 24 h. Twelve healthy subjects (19 to 23 years of age) were asked to drink approximately 50 mL of unsweetened tea from a plastic bottle. The mouths of the bottles were swabbed with sterile cotton, and the swabs and the remaining tea in the bottles were analyzed by anaerobic culture and 16S rRNA gene sequencing. Metagenomic analysis of the 16S rRNA gene was also performed. The mean amounts of bacteria were (1.8 ± 1.7) × 104 colony-forming units (CFU)/mL and (1.4 ± 1.5) × 104 CFU/mL at the mouth of the bottles immediately after and 24 h after drinking, respectively. In contrast, (0.8 ± 1.6) × 104 CFU/mL and (2.5 ± 2.6) × 106 CFU/mL were recovered from the remaining tea immediately after and 24 h after drinking, respectively. Streptococcus (59.9%) were predominant at the mouth of the bottles immediately after drinking, followed by Schaalia (5.5%), Gemella (5.5%), Actinomyces (4.9%), Cutibacterium (4.9%), and Veillonella (3.6%); the culture and metagenomic analyses showed similar findings for the major species of detected bacteria, including Streptococcus (59.9%, and 10.711%), Neisseria (1.6%, and 24.245%), Haemophilus (0.6%, and 15.658%), Gemella (5.5%, and 0.381%), Cutibacterium (4.9%, and 0.041%), Rothia (2.6%, and 4.170%), Veillonella (3.6%, and 1.130%), Actinomyces (4.9%, and 0.406%), Prevotella (1.6%, and 0.442%), Fusobacterium (1.0%, and 0.461%), Capnocytophaga (0.3%, and 0.028%), and Porphyromonas (1.0%, and 0.060%), respectively. Furthermore, Streptococcus were the most commonly detected bacteria 24 h after drinking. These findings demonstrated that oral bacteria were present at the mouth of the bottles and in the remaining tea after drinking.

DOI： 10.3390/dj9060058

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DOI： 10.1016/j.job.2021.03.001

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This longitudinal study aimed to clarify the relationship of oral health in infancy with that in adulthood among participants who were the subjects of the oral health promotion project (OHPP) conducted in Miyako Island, Okinawa Prefecture, Japan, since 1984. Twenty-seven subjects, around 35 years of age, were examined for dental caries, periodontal diseases (community periodontal index), dental plaque, occlusion, and bite-force and compared with those at 4 and 13-15 years of age. The dental caries status and maximum bite force in adulthood was significantly reflected for those at 4 and 13-15 years of age (p < 0.05). CPI in adulthood was related to the dental caries status at 4 and 13-15 years of age but not to the gingival score at 4 years of age, and it was weakly related to the gingival score at 13-15 years (r = 0.264, p > 0.05). Most of the normal occlusion at 4 years of age became normal permanent occlusion in adulthood (88.9%). Most of the cases involving the discrepancy factor retained the same condition in both the deciduous and permanent dentitions (83.3%) (p < 0.001). Those who participated in the OHPP soon after birth showed significantly fewer DMFT (p < 0.05) compared with those who did not. This study revealed that oral health at 4 years of age was related to that in adulthood, suggesting that fostering good oral health soon after birth is of great importance.

DOI： 10.3390/dj9020017

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© 2020 The Japanese Society for Regenerative Medicine Introduction: Responses of oral-microflora-exposed dental pulp to a triple antibiotic paste (TAP), a mixture of ciprofloxacin, metronidazole, and minocycline in ointment with macrogol and propylene glycol, remain to be fully clarified at the cellular level. This study aimed to elucidate responses of oral-microflora-exposed dental pulp to capping with TAP in mouse molars. Methods: A cavity was prepared on the first molars of 6-week-old mice to expose the dental pulp for 24 h. The exposed pulp was capped with TAP (TAP group) or calcium hydroxide cement (CH group), in addition to the combination of macrogol (M) and propylene glycol (P) (MP, control group), followed by a glass ionomer cement filling. The samples were collected at intervals of 1, 2, and 3 weeks, and immunohistochemistry for nestin and Ki-67 and deoxyuride-5′-triphosphate biotin nick end labeling (TUNEL) assay were performed in addition to quantitative real-time polymerase chain reaction (qRT-PCR) analyses. Results: The highest occurrence rate of pulp necrosis was found in the control group followed by the CH group at Weeks 2 and 3, whereas the highest occurrence rate of healed areas in the dental pulp was observed in the TAP group at each time point. Tertiary dentin formation was first observed in the dental pulp of the TAP group at Week 2. In contrast, bone-like and/or fibrous tissues were frequently observed in the CH group. qRT-PCR analyses clarified that TAP activated the stem and dendritic cells at Weeks 1 and 2, respectively. Conclusions: The use of TAP as a pulp-capping agent improved the healing process of oral-microflora-exposed dental pulp in mouse molars.

DOI： 10.1016/j.reth.2020.10.001

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Recently, it was suggested that the nitrite (NO2-) produced from NO3- by oral bacteria might contribute to oral and general health. Therefore, we aimed to clarify the detailed information about the bacterial NO2-production in the oral biofilm. Dental plaque and tongue-coating samples were collected, then the NO2-producing activity was measured. Furthermore, the composition of the NO2--producing bacterial population were identified using the Griess reagent-containing agar overlay method and molecular biological method. NO2--producing activity per mg wet weight varied among individuals but was higher in dental plaque. Additionally, anaerobic bacteria exhibited higher numbers of NO2--producing bacteria, except in the adults' dental plaque. The proportion of NO2--producing bacteria also varied among individuals, but a positive correlation was found between NO2--producing activity and the number of NO2--producing bacteria, especially in dental plaque. Overall, the major NO2--producing bacteria were identified as Actinomyces, Schaalia, Veillonella and Neisseria. Furthermore, Rothia was specifically detected in the tongue coatings of children. These results suggest that dental plaque has higher NO2--producing activity and that this activity depends not on the presence of specific bacteria or the bacterial compositions, but on the number of NO2--producing bacteria, although interindividual differences were detected.

DOI： 10.1038/s41598-020-73479-1

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This study evaluated the effectiveness of a life-enhancement program designed to focus on dining conditions in welfare facilities for seniors living in Japan. Effectiveness was specifically evaluated based on whether improvements were achieved in (1) nutritional status, (2) oral health, (3) frequency of fever, and (4) vitality of appetite across three sites. As part of a comprehensive-care initiative that began with dining support, the program consisted of two main components: (1) a 3-month intensive program comprised of (a) collective experiential learning for residents and staff (including nutritionists, nurses, and physiotherapists) and (b) a tailor-made individual program for residents followed by (2) a 3-month continuation program. Participants included 168 individuals (31 males and 137 females) from a total of three facilities (average age was 85.9 [60-104] years). Results showed that the intensive program significantly improved nutritional status (e.g., BMI, caloric intake, and water intake; P < 0.000-0.005) and tongue movement (P < 0.000) while significantly reducing dental-plaque and tongue-coating indices (P < 0.000). Significant improvements were also achieved for degree of appetite and vitality indices (P < 0.000-0.001). However, incidences of fever were not reduced. These findings indicate that the program effectively improved nutritional status, oral health, vitality, and appetite. However, these effects did not sufficiently remain once the program was finished, thus suggesting the need for a continuous intervention.

DOI： 10.31372/20200502.1089

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The aim of this study is to determine the effect of food consistency on health and related factors among residents in welfare facilities for seniors (n = 227; mean age, 86.2 ± 8.0 years; 78.9% female). Residents who ate regular food had a lower incidence of fever during the 3-month period (p < 0.001) and consumed more calories (1325.97 ± 220.2 kcal) than those who ate chopped (1125.0 ± 256.8 kcal), paste (1122.0 ± 288.5 kcal), and gastric tube food (812.5 ± 150.7 kcal) (p < 0.001). Modifying a resident's food by making it softer and finer did not reduce the incidence of choking. Logistic regression analysis (backward elimination method) revealed four factors related to eating regular food: vitality index, appetite, number of remaining teeth, and choking frequency. Causal relationships were not obtained because this was a cross-sectional study. The findings of this study suggest that a regular consistency of food positively influences the health of older individuals.

DOI： 10.3390/dj8010009

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© 2019 Mashima et al. To date, Veillonella tobetsuensis has been known as an oral anaerobe and a facilitator of early-stage oral biofilm development with streptococci. Here, we report the draft genome sequences of 2 strains of V. tobetsuensis first isolated from intraoperative bronchial fluids of elderly patients with pulmonary carcinoma.

DOI： 10.1128/MRA.00397-19

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It is suspected that oral bacteria are transferred to the liquid baby formula through the artificial nipple and multiply in the bottle after feeding. In the present study, in order to understand the influence of bacteria on liquid baby formula after feeding, the transfer of oral bacteria through artificial nipples and their survival in liquid baby formula were examined immediately after drinking as well as after storage at 4°C for 3 h. Four healthy human subjects (20-23 years old) were asked to drink liquid baby formula (Aptamil®, ca. 50 mL) from baby bottles using artificial nipples. Samples of the liquid baby formula (immediately after drinking and 3 h later) were inoculated onto blood agar plates and incubated anaerobically at 37°C for 7 days. Salivary samples from each subject and 6 newborn infants were also cultured. Genomic DNA was extracted from individual colonies, and bacterial species were identified by 16S rRNA gene sequencing. The mean amounts of bacteria (CFU/mL) were (3.2 ± 3.0) ×104 and (3.4 ± 3.3) ×104 immediately after drinking and 3 h later, respectively. Streptococcus (41.6 and 40.5%), Actinomyces (24.3 and 21.5%) and Veillonella (16.2 and 11.0%) were recovered from the samples immediately after drinking and 3 h later, respectively. On the other hand, Streptococcus (38.9%), Actinomyces (17.1%), Neisseria (9.1%), Prevotella (6.9%), Rothia (6.9%) and Gemella (5.1%) were predominant in the saliva of adult subjects, and Streptococcus (65.2%), Staphylococcus (18.5%), Gemella (8.2%) and Rothia (5.4%) were predominant in the saliva of infant subjects. From these findings, oral bacteria, e.g., Streptococcus, Gemella and Rothia, were found to transfer into the liquid baby formula through artificial nipples, and the bacterial composition in the remaining liquid baby formula was found to resemble that of human saliva. The bacterial levels were similar between immediately after drinking and when stored at 4°C for 3 h, suggesting that the remaining liquid baby formula may be preserved in a refrigerator for a specified amount of time.

DOI： 10.2220/biomedres.40.163

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Language：English   Publishing type：Research paper (scientific journal)  

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© 2019, Biomedical Research Foundation. All rights reserved. One of the most severe complications of lung resection is postoperative pneumonia, and its prevention and prediction are critical. Exhaled acetone and isoprene are thought to be related to metabolism; however, little is known on their relationship with bacteria living in the oral cavity or their meaning in the acute phase in perioperative lung cancer patients. We measured acetone and isoprene in exhaled breath of 13 Japanese patients with lung cancer (3 women and 10 men, age range 62–82 years, mean 72.4 years) before breakfast during hospitalization, and compared with two acute-phase proteins, C-reactive protein (CRP) and albumin in blood serum, as well as the total number of bacteria in saliva and their activity to produce acetone and isoprene. Before operation, intensive oral care was carried out for each patient to prevent postoperative pneumonia, and swallowing and cough reflexes were measured for 12 of 13 patients to assess risk of postoperative pneumonia. Breath and saliva were sampled before intensive oral care (T1), after oral care but before operation (T2), and after operation (T3) during hospitalization. The total number of oral bacteria in saliva decreased significantly from T1 to T2 among 13 patients. No acetone or isoprene was detected from saliva after in vitro incubation under anaerobic or aerobic conditions, but both acetone and isoprene were detected in breath. After operation, breath acetone correlated significantly with CRP (Spearman’s ρ = 0.559, P = 0.03), but not with albumin. Breath isoprene correlated significantly with albumin (Spearman’s ρ = 0.659, P = 0.008), but not with CRP after operation. Although the number of subjects was small, our results support the hypothesis that breath acetone and isoprene may be related with these acute-phase proteins, which reflect inflammatory reactions and subsequent changes in metabolism in the early postoperative phase of lung resection.

DOI： 10.2220/biomedres.40.29

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：TAYLOR & FRANCIS INC  

Objectives: A program fostering self-management for the elderly was implemented and the effects of the program and their continuities were assessed. Methods: Subjects consisted of 19 males and 131 females (average age, 73.1 +/- 7.4; range, 60-94years). The intervention program consisted of the collective experience learning and private consultation. The collective experience learnings included; (1) monitoring the oral condition and practicing oral self-care, (2) monitoring the oral function and practicing oral exercises, and (3) group discussion on continuing oral self-care. Outcomes were evaluated at the beginning and the end of the intervention program, and three months after the investigation by the scores in; (1) oral self-care (2) oral condition, i.e., decayed teeth, community periodontal index (CPI), deposits of plaque and dental calculus, (3) oral function such as RSST, oral diadochokinesis, (4) QOL (SF-8 v2, and GOHAI), and (5) cognitive function (MMSE-J). Informed consent was obtained from all subjects, and this study was approved by the Research Ethics Committee of the University of Hyogo. Results and Discussion: At three months after intervention, 124 subjects continued participating and 88 subjects (71%) completed all data. On the oral self-care, subjects cleaned their teeth more often and longer than before (P &lt; 0.001). The use of dental floss and interdental brushing significantly increased in number (P &lt; 0.001), and 67 participants (54%) visited the dentist during the program. CPI and deposits of plaque were significantly reduced after intervention (P &lt; 0.001). The scores of oral function also significantly improved (P &lt; 0.001-0.05). The scores of QOL (physical health), oral QOL and cognitive function significantly improved (P &lt; 0.001-0.05). These results suggest that this program not only promotes oral self-care, resulting in good oral health conditions, but also improves cognitive functions of the elderly.

DOI： 10.1080/10798587.2017.1348459

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Japanese Association for Oral Biology  

Objectives The frequency of periodontitis among elderly is increasing in Japan. This study aimed to quantify the periodontitis-associated bacteria in subgingival plaques from elderly patients with periodontitis and from periodontally healthy subjects. Methods Subgingival plaque samples were collected from independent subjects (mean age=71.1 years, n=518). All samples were subjected to real-time PCR analysis to assess the presence of Porphyromonas gingivalis. In addition, we tested for Tannerella forsythia, Eubacterium saphenum and Streptococcus oralis in the earliest 95 samples
and performed fimA gene classification of P. gingivalis in the latest 49 samples. Results P. gingivalis and T. forsythia comprised a significantly higher proportion of total bacteria in subjects with periodontitis (1.1% and 5.1%) than periodontally healthy subjects (0.3% and 1.4%, respectively). The proportion of E. saphenum was low in both groups, whereas that of S. oralis was higher in periodontally healthy subjects. In 24 of 49 samples, fimA genotypes were detected and classified. Genotypes Ib (n=5) and II (n=7) were identified in those of subjects with periodontitis (n=15)
while those of healthy subjects (n=9) were found to belong to genotypes I (n=2), II (n=2), III (n=2) and IV (n=3). In 4 out of the 5 subjects in whom P. gingivalis was detected at healthy sites, the fimA genotypes were identical between periodontitis and healthy sites, but the mean proportion of P. gingivalis was significantly higher at periodontitis sites (3.0%) than at healthy sites (0.5%) (P&lt
0.05). Conclusions This study suggests that an increase in P. gingivalis and T. forsythia may be associated with periodontitis in the elderly, and we have identified characteristic pathogenic fimA genotypes that target this vulnerable group.

DOI： 10.1016/j.job.2015.12.002

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：BIOMEDICAL RESEARCH PRESS LTD  

The onset of plaque-mediated disease, including dental caries and periodontal diseases, is highly associated with compositional change of the resident microflora from the ecological perspective. As specific bacterial profiles have been linked to different disease stages, microbial compositional measurements might therefore have great value for clinical diagnosis. Previously we have reported a dry-reagent strip biosensor-PCR-dipstick DNA chromatography, which utilized molecular recognition of oligonucleotides and biotin-streptavidin, and the optical property of colored microspheres, for semiquantifying a five-membered subgroup of caries-associated bacterial species in supragingival plaque from healthy coronal surfaces of teeth. The present study aimed to evaluate this technique's ability to differentiate microflora by comparing the subset profiles. Sixteen subgingival plaque specimens were pooled from periodontal pockets and analyzed for the composition of Streptococcus mutans, Streptococcus sobrinus, Scardovia wiggsiae, Actinomyces sp. and Veillonella parvula. Detection frequencies, relative abundance of each bacterial species, and the five-membered bacterial profiles were compared between supra- and subgingival groups. The supragingival plaque harbored significantly more of the tested species and higher amount of Actinomyces sp. and V. parvula. In subgingival plaque, the predominance was obscured, since several highly overlapped profiles were found at comparable frequencies. Thus, PCR-dipstick DNA chromatography using the same plaque sample enabled simultaneous profiling of multiple species at species level and facilitated discrimination between anticipated different microflora, making this technique a promising chair-side microbiota profiling method.

DOI： 10.2220/biomedres.37.29

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (international conference proceedings)  

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：TOHOKU UNIV MEDICAL PRESS  

Porphyromonas strains, including Porphyromonas-like strains, have been isolated from oral and various other systemic infections. The characterization of such strains is a crucial issue, because such information contributes to both the taxonomy of anaerobic bacteria and the clinical aspects of infectious diseases. We previously isolated four Porphyromonas-like strains from intraoperative bronchial fluids of a patient with non-small cell lung cancer. This study aimed to characterize the genetic, biochemical and chemotaxonomic aspects of these isolates. Each strain only grew under anaerobic conditions and their colony morphology was convex, 0.1-1.0 mm in diameter, light gray, and slightly glistening colony, with no black or brown pigmentation on blood agar plates after five-day incubation. The pigmentation was helpful to differentiate the isolates from other Porphyromonas, as most of Porphyromonas species show the pigmentation. In the 16S rRNA gene phylogenetic analysis (98% sequence identity of isolates indicates the same species), the four isolates were closely related to one another (99.7-100.0%), but not related to Porphyromonas (P.) catoniae, the closest species (96.9%). In addition, the DNA-DNA hybridization data revealed less than 16% similarity values between a representative isolate and the P. catoniae, indicating that the strains were genetically independent. Biochemically, the isolates could be differentiated from closely related species, i.e., P. catoniae, P. gingivalis, P. gulae, and P. pogonae, with trypsin activity (negative only in the isolates) and leucine arylamidase activity (positive only in the isolates). We therefore propose a new species to include these isolates: Porphyromonas bronchialis sp. nov.

DOI： 10.1620/tjem.237.31

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1679/aohc.73.165

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Japanese Association for Oral Biology  

Objectives The source of the bacteria involved in silent aspiration remains to be completely defined. This study aimed to obtain reliable evidence on silent aspiration of oral bacteria in elderly patients. Methods After obtaining informed consent, the cough and swallowing reflexes of patients were assessed. Bronchial fluids from patients undergoing lung resections were collected with a micro-sampling probe, and α-amylase activity of bronchial fluids was measured to estimate the degree of silent aspiration. The bronchial fluids were cultured aerobically and anaerobically on blood agar plates, and colonies were identified by 16S rRNA gene sequencing. Additionally, whole saliva bacterial amounts and composition were analyzed. Results Six patients (72.2±5.8 years) exhibited an impaired swallowing reflex and 5 (75.4±7.9 years) had a normal swallowing reflex, while all patients had a normal cough reflex. α-Amylase activity was detected in bronchial fluids of both the impaired and normal reflex groups. The amount of anaerobic bacteria in bronchial fluids in the impaired reflex group [(3.0±3.5)×104] was higher than in the normal reflex group [(2.5±5.3)×104sup], although the difference was not significant. Actinomyces, Gemella, Streptococcus, Rothia, Mogibacterium, and Campylobacter were the predominant bacterial species in bronchial fluids of the impaired reflex group, while Streptococcus, Lactobacillus, Veillonella, and Actinomyces were predominant in the normal reflex group. Conclusions Our results suggest that bacteria in bronchial fluids associated with silent aspiration are derived from saliva, and that the bronchial fluids of elderly patients with an impaired swallowing reflex may have a characteristic microbiota.

DOI： 10.1016/j.job.2014.11.001

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER GMBH, URBAN & FISCHER VERLAG  

During the process of identifying a Gram-negative coccobacillus isolated from a human clinical specimen, we found that the isolate's 16S rRNA gene had very close sequence identity with that of a variant Porphyromonas isolated from polymicrobial infections in the central bearded dragon, a species of lizard [2]. The 16S rRNA gene sequences of the human isolate and of six isolates from lizards were nearly identical (99.9-100%). Phylogenetic analysis placed all of these isolates in a single phylogenetic cluster well separated from other species in the genus Porphyromonas. The closest species was Porphyromoncts catoniae with 90.7-90.9% sequence identity, although there was less than 6% DNA similarity between the P. catoniae type strain and our representative isolates from lizards (PAGU 1787(T)) and human (PAGU 1776). These isolates could grow under anaerobic or microaerobic conditions (6% 02 atmosphere). The isolates were positive for catalase and very strong beta-hemolytic activity, but did not show black or brown pigmentation. Biochemically, the isolates could be differentiated from closely related species by pyroglutamic acid arylamidase and glycine arylamidase activity, and some others. The fermentation products mainly included succinic acid and propionic acid. The major fatty acids detected in cells of the isolates were iso-C15:0, anteiso-C15:0, and 3OH-iso-C17:0. The G + C content was 43.0 +/- 0.62 mol%. The species name Porphyromonas pogonae sp. nov. is proposed for these isolates with the type strain of PAGU 1787(T) (=MI 10-1288(T) = JCM 19732(T) = ATCC BAA-2643(T)). (C) 2014 Elsevier GmbH. All rights reserved.

DOI： 10.1016/j.syapm.2014.11.004

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Japanese Association for Oral Biology  

Background Since most oral malodor originates from microbial activities in the mouth, the role of microorganisms in producing malodorous gases has been clarified by numerous studies, accompanied by the development of analytical techniques for treatment of halitosis. Highlight Oral microorganisms should be controlled to prevent aspiration pneumonia, especially for elderly perioperative patients. Malodorous gases from the mouth can be an indicator of oral or systemic conditions among patients in intensive care units. Recently, malodorous gases originating from oral microorganisms have been reported as a causal factor in carcinogenesis. Conclusion Further analysis of oral malodor might be useful in accessing the risk of aspiration pneumonia and oral cancer.

DOI： 10.1016/j.job.2015.05.004

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

Postoperative pneumonia may occur when upper respiratory tract protective reflexes such as cough and/or swallowing reflexes are impaired; thus, silent aspiration of oral bacteria may be a causative factor in postoperative pneumonia. This study aimed to quantify and identify bacteria in intraoperative bronchial fluids and to evaluate the relationship between impairment of cough/swallowing reflexes and silent aspiration of oral bacteria in elderly patients. After obtaining informed consent, cough and swallowing reflexes were assessed using an ultrasonic nebulizer and a nasal catheter, respectively. Using a micro-sampling probe, intraoperative bronchial fluids were collected from nine subjects with pulmonary carcinoma and cultured anaerobically on blood agar plates. After 7 days, CFUs were counted and isolated bacteria were identified by 16S rRNA gene sequencing. Four subjects (aged 71.0 +/- 8.4 years) had impaired swallowing reflexes with normal cough reflexes, whereas five subjects (73.6 +/- 6.5 years) had normal cough and swallowing reflexes. The bacterial counts (mean CFU +/- SD) tended to be higher in intraoperative bronchial fluids of subjects with impaired swallowing reflexes ([5.1 +/- 7.7] x 10(5)) than in those of subjects with normal reflexes ([1.2 +/- 1.9] x 10(5)); however, this difference was not statistically significant. Predominant isolates from intraoperative bronchial fluids were Streptococcus (41.8%), Veillonella (11.4%), Gemella (8.9%), Porphyromonas (7.6%), Olsenella (6.3%) and Eikenella (6.3%). These findings indicate that intraoperative bronchial fluids contain bacteria, probably derived from the oral microbiota, and suggest that silent aspiration of oral bacteria occurs in elderly patients irrespective of impairment of swallowing reflex.

DOI： 10.1111/1348-0421.12157

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：HINDAWI PUBLISHING CORPORATION  

A complex of species has been associated with dental caries under the ecological hypothesis. This study aimed to develop a rapid, sensitive PCR-dipstick DNA chromatography assay that could be read by eye for multiplex and semiquantitative analysis of plaque bacteria. Parallel oligonucleotides were immobilized on a dipstick strip for multiplex analysis of target DNA sequences of the cariesassociated bacteria, Streptococcus mutans, Streptococcus sobrinus, Scardovia wiggsiae, Actinomyces species, and Veillonella parvula. Streptavidin-coated blue-colored latex microspheres were to generate signal. Target DNA amplicons with an oligonucleotide-tagged terminus and a biotinylated terminus were coupled with latex beads through a streptavidin-biotin interaction and then hybridized with complementary oligonucleotides on the strip. The accumulation of captured latex beads on the test and control lines produced blue bands, enabling visual detection with the naked eye. The PCR-dipstick DNA chromatography detected quantities as low as 100 pg of DNA amplicons and demonstrated 10-to 1000-fold higher sensitivity than PCR-agarose gel electrophoresis, depending on the target bacterial species. Semiquantification of bacteria was performed by obtaining a series of chromatograms using serial 10-fold dilution of PCR-amplified DNA extracted fromdental plaque samples. The assay time was less than 3 h. The semiquantification procedure revealed the relative amounts of each test species in dental plaque samples, indicating that this disposable device has great potential in analysis of microbial composition in the oral cavity and intestinal tract, as well as in point-of-care diagnosis of microbiota-associated diseases.

DOI： 10.1155/2014/180323

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Purpose: To quantify and identify bacteria detected in acrylic resin dentures and dento-maxillary obturator-prostheses after long-term use. Methods: The internal layer of denture bases from 13 daily-use removable acrylic resin dentures was sampled, while the inner fluid samples/no-fluid samples of obturators were collected from II in-use acrylic resin dento-maxillary obturator-prostheses. Samples were cultured, and isolated bacteria were counted and identified by molecular biological methods. Results: Bacteria were detected in five (38.5%) acrylic resin dentures and six (54.5%) acrylic resin obturators. Four Lactobacillus species and one Propionibacterium species were isolated from three repaired denture bases, and from two non-repaired dentures, two Actinomyces species and Streptococcus mutans were isolated. On the other hand, 17 bacterial species, belonging to the family and genera of Olsenella, Bacillus, Citrobacter, Entembacteriaceae, Lactobacillus, Pantoea, Peptoniphilus, Klebsiella and Pseudomonas, were isolated from obturators. Several species of viable bacteria were detected in acrylic resin denture bases and obturators. (Am J Dent 2012;25:171-175).

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：MICROBIOLOGICAL SOCIETY KOREA  

This study aimed to profile the microflora in infected root canals before and after root canal treatment using culture-independent methods. Six infected root canals in single-rooted teeth with periapical lesions from five subjects were included. Quantification of total bacteria was performed by real-time PCR with primers targeting 16S rRNA genes. PCR products with universal 16S rRNA gene primers were cloned and partially sequenced, and bacterial identification at the species level was performed by comparative analysis with the GenBank database. The concentration of extracted DNA before treatment was higher than that after root canal treatment, although the difference was not statistically significant. Sequence analysis revealed that oral bacteria such as Fusobacterium, Streptococcus, Olsenella, and Pseudoramibacter detected in cases before root canal treatment disappeared after treatment. These results suggest that the root canal microflora are distinct before and after root canal treatment, and that treatment changes the microflora in both quantity and quality.

DOI： 10.1007/s12275-012-0459-4

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Objective. The bacterial examination has been performed during the course of the root canal treatment. In the present pilot study, the new developed method, using fluorescence reagents and a membrane filter, was applied to the detection and quantification of bacteria in infected root canals, in order to evaluate the outcomes of the treatment. Methods. Six infected root canals with periapical lesions from 5 subjects were included. Informed consent was obtained from all subjects (age ranges, 23-79 years). Samples from infected root canals were collected at the beginning of the treatment (termed #25 First), the end of the first day of treatment (termed #55 First), and the next appointment day (termed #55 Second). Then, the bacterial count (CFU) was measured using fluorescence reagents (4′,6′-diamidino-2-phenylindole and propidium iodide) and the polycarbonate membrane filter by Bioplorer. Results. The mean SD of CFU in the sample of #25 First was (1.0 ± 1.4) 10 5. As the root canal treatment progressed, the CFU decreased as 7.9 × 10 3 (#55 First) and 4.3 × 10 2 (#55 Second). Conclusion. In the present pilot study, rapid detection and quantification of bacteria in infected root canals were found to be successfully performed using fluorescence reagents and a membrane filter (Bioplorer analysis). © 2012 Takuichi Sato et al.

DOI： 10.1155/2012/172935

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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) 10 6 (range 8.0 × 10 1 - 3.1 × 10 6), 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. © Copyright 2012 Takuichi Sato et al.

DOI： 10.1155/2012/609689

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Purpose: To profile plaque microflora on root-caries lesions, and to examine the protein-denaturing activity as a pilot study. Methods: Six subjects with root-caries were investigated. Plaque samples on root caries lesions (R), as well as from healthy supragingival sites (S) and periodontal pockets (&gt;= 5 mm) (P) were collected and cultured anaerobically on blood agar plates. The isolated bacteria were identified by 16S rRNA sequencing analysis, and examined for the protein-denaturing activity using the skim-milk plates and the SDS-PAGE, and for the acidogenicity using the FAB broth containing 1% glucose. Results: Propionibacterium, Actinomyces, Streptococcus, Lactobacillus and Bifidobacterium were predominant in R, while Actinomyces, Streptococcus, Veillonella and Capnocytophaga in S, and Actinomyces, Prevotella, Actinobaculum, Streptococcus, Olsenella and Eubacterium were predominant in P. Proteolytic bacteria comprised 40%, 26% and 57% of microflora in R, S and P. respectively. The skim-milk plates distinguished between protein-degrading and protein-coagulating bacteria, which comprised 7 and 33%, 0 and 26%, and 17 and 40% of microflora, in R, S and P, respectively. The SDS-PAGE analysis revealed that protein-degrading isolates were capable of degrading collagen molecules. Furthermore, the final culture pHs of protein-degrading and -coagulating bacteria were 5.0-5.4 and 3.8-3.9, respectively. The latter pH was low enough to denature proteins in skim milk. The microbial composition of R was distinct from those of S and P. (Am J Dent 2011;24:295-299).

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Candida species were detected and identified in samples from the buccal mucosa, dorsal surface of the tongue and supragingival plaque of subjects with oral lichen planus (OLP). The Candida in the samples were cultured on selection agars, and identified by sequence analyses of 18S, 5.8S and 25/28S rRNA. The isolation frequency of Candida was higher in subjects with OLP than in those with healthy oral mucosa. Non-C. albicans were only isolated from people with OLP. These results support the notion that subjects with OLP are more likely to have oral colonization with Candida, and that non-C. albicans are specifically present in subjects with this condition.

DOI： 10.1111/j.1348-0421.2010.00285.x

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Authorship：Last author   Language：Japanese   Publishing type：Research paper (other academic)   Publisher：公益社団法人日本薬学会  

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Background and Objective: Leukocytes and epithelium are the first line of defense in preventing bacterial invasion into periodontium. Some of these cells die in gingival crevicular fluid, whereupon their DNA is spilled out. The present study was designed to investigate the profile of host beta-globin gene fragments in the gingival crevicular fluid of various periodontal conditions.
Material and Methods: Gingival crevicular fluid from 40 teeth with chronic periodontitis, 30 with gingivitis and 22 that were clinically healthy were centrifuged (3000g, 10 min). The supernatant (cell-free gingival crevicular fluid) was centrifuged again (13,000g, 10 min), resulting in the pellet and the supernatant as debris and debris-free fractions, respectively. Specific primers for amplifying 110 bp, 536 bp and 2 kb amplicons of human beta-globin gene were used to investigate host DNA by quantitative and qualitative polymerase chain reaction.
Results: The periodontitis group showed the largest amount of host beta-globin gene fragments, while the healthy group had the lowest. In the debris and debris-free fractions, the 536 bp and 2 kb amplicons were more often detected in the periodontitis group than in the other groups. Interestingly, the presence of 2 kb amplicon in the debris fraction could be used to discriminate periodontitis from gingivitis and healthy groups because we found it in 85% of periodontitis samples but only in 13% of gingivitis samples, and it was absent in the healthy group.
Conclusion: This study shows the different DNA profiles of cell-free gingival crevicular fluid in periodontal health and disease. It suggests that the quantity and quality of host DNA are dependent on the disease conditions. Therefore, the beta-globin gene fragments in cell-free gingival crevicular fluid may be a potential biomarker of periodontal disease progression.

DOI： 10.1111/j.1600-0765.2008.01197.x

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DOI： 10.1111/j.1600-0501.2006.01294.x

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A disintegrin and metalloproteinase with thrombospondin motif (adamalysin-thrombospondins, ADAMTS) degrades aggrecan, one of the major extracellular matrix (ECM) components in cartilage. Mandibular condylar cartilage differs from primary cartilage, such as articular and growth plate cartilage, in its metabolism of ECM, proliferation, and differentiation. Mandibular condylar cartilage acts as both articular and growth plate cartilage in the growing period, while it remains as articular cartilage after growth. We hypothesized that functional and ECM differences between condylar and primary cartilages give rise to differences in gene expression patterns and levels of aggrecan and ADAMTS-1, -4, and -5 during growth and aging. We employed in situ hybridization and semiquantitative RT-PCR to identify mRNA expression for these molecules in condylar cartilage and primary cartilages during growth and aging. All of the ADAMTSs presented characteristic, age-dependent expression patterns and levels among the cartilages tested in this study. ADAMTS-5 mainly contributed to ECM metabolism in growth plate and condylar cartilage during growth. ADAMTS-1 and ADAMTS-4 may be involved in ECM turn over in articular cartilage. The results of the present study reveal that ECM metabolism and expression of related proteolytic enzymes in primary and secondary cartilages may be differentially regulated during growth and aging.

DOI： 10.1007/s00418-006-0171-8

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Background/aims: Postoperative maxillary cyst (POMC) is known to occur as a delayed complication of radical maxillary sinus surgery, such as Caldwell-Luc surgery. The cyst gradually expands with no symptoms over a period of years, and then occasionally causes swelling and pain in the buccal region and/or the mucogingival fold. It is probable that bacterial infection affects the progression of POMC symptoms. The aims of this study were to determine the bacterial density and to examine the presence of 20 oral bacteria in POMC fluids.
Methods: POMC fluids (4 purulent, 2 mucous and 4 serous) were sampled from 10 subjects (aged 43-77 years). Bacterial quantification and detection were performed by real-time polymerase chain reaction (PCR) and nested PCR based on bacterial 16S rRNA genes, respectively.
Results: Bacterial DNA was detected in all samples and the average concentrations of bacterial DNA were 5.9 (purulent), 0.5 (mucous), and 0.7 (serous) ng/mg of sample. Twelve bacterial species, including anginosus streptococci, known to be associated with abscess formation, were detected in the purulent fluids, while two and five species were detected in the mucous and serous fluids, respectively.
Conclusion: Purulent fluids contained numerous bacteria of various types, thus suggesting that oral bacteria may cause symptoms such as pain in POMC with purulent fluids. Mucous and serous fluids also contained bacteria, although their numbers were small, thus suggesting an association between bacteria and progression of POMC.

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The aims of this study were to identify hydrogen sulfide (H2S)-producing bacteria among tongue biofilm microflora and to investigate the relationship between bacterial flora and H2S levels in mouth air. Oral malodour levels in 10 subjects (age 21-56 years) were assessed by gas chromatography, and Breathtron and organoleptic scores. Based on these assessments, subjects were divided into two groups: an odour group and a no/low odour group. Tongue coatings were sampled and spread onto Fastidious Anaerobe Agar plates containing 0(.)05 % cysteine, 0(.)12 % glutathione and 0(.)02 % lead acetate, and were then incubated anaerobically at 37 degrees C for 2 weeks. Bacteria forming black or grey colonies were selected as H2S-producing phenotypes. The numbers of total bacteria (P &lt; 0.005) and H2S-producing bacteria (P &lt; 0.05) in the odour group were significantly larger than those in the no/low odour group. Bacteria forming black or grey colonies (1126 isolates from the odour group; 242 isolates from the no/low odour group) were subcultured, confirmed as producing H2S and identified according to 16S rRNA gene sequencing. Species of Veillonella (38.1 % in odour group; 46.3 % in no/low odour group), Actinomyces (25.4 %; 17.7 %) and Prevotella (10.3 %; 7.8 %) were the predominant H2S-producing bacteria in both the odour and no/low odour groups. These results suggest that an increase in the number of H2S-producing bacteria in the tongue biofilm is responsible for oral malodour, although the bacterial composition of tongue biofilm was similar between the two groups.

DOI： 10.1099/jmm.0.46118-0

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The aim of this study was to compare the detection frequencies of 25 bacterial species in subgingival and supragingival plaque of 18 untreated periodontitis subjects and 12 periodontally healthy subjects. Genomic DNA was extracted from subgingival and supragingival plaque samples, and bacterial detection was performed by polymerase chain reaction of the 16S rRNA genes. Fourteen bacteria showed no relationship with periodontitis, and 11 of these 14 species were frequently detected (greater than or equal to50%) in subgingival plaque in both periodontitis and healthy subjects. Nine bacteria such as Eubacterium saphenum, Prevotella intermedia, and Treponema denticola seemed to be related to periodontitis; their detection frequencies in subgingival plaque samples were higher in periodontitis than in healthy subjects, but these differences were not statistically significant by multiple comparisons (0.002less than or equal toP&lt;0.05). Two species (Mogibacterium timidum and Porphyromonas gingivalis) were detected significantly more frequently in subgingival plaque of periodontitis subjects than of healthy subjects (P&lt;0.002), with P. gingivalis being detected only in periodontitis subjects, suggesting that these two species are closely related to periodontitis. There were no significant differences in the detection frequencies of the 25 bacteria between subgingival and supragingival plaque, suggesting that the bacterial flora of supragingival plaque reflects that of subgingival plaque.

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The Patched (PTC) gene is responsible for basal cell nevus syndrome (BCNS) accompanied by multiple odontogenic keratocysts (OKCs), and its product plays a role in the Sonic hedgehog (SHH) signaling pathway involving smoothened (SMO) and GLI-1. To clarify the role of SHH signaling in OKCs, the expression of SHH, PTC, SMO, and GLI-1 and mutations of PTC were examined in 18 sporadic, 4 BCNS-associated OKCs and 7 control gingivae. SHH, PTC, SMO, and GLI-1 were detected in all OKC and gingiva samples by reverse transcriptase-polymerase chain reaction (RT-PCR). Immunoreactivity for SHH and GLI-1 was markedly higher in epithelial components than in subepithelial cells, while immunoreactivity for PTC and SMO was similar in epithelial components and subepithelial cells in OKCs. The positive rate of PTC and SMO expression in subepithelial cells of OKCs was significantly higher than that in gingivae. The positive rate of GLI-1 expression in subepithelial cells of BCNS-associated OKCs was significantly higher than that in primary OKCs. These results suggest that the SHH signaling might be involved in the pathophysiologic nature of OKCs. While mutations of the PTC gene could not be detected in 4 BCNS-associated OKCs by direct DNA sequencing, 3 of 5 primary and 4 of 4 recurrent OKCs had several mutations of this gene. These results suggest that PTC mutations are probably related not only to BCNS-associated OKCs but also to sporadic OKCs.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：KARGER  

This study was carried out to develop a method for mapping the distribution of cariogenic oral streptococci, Streptococcus mutans and Streptococcus sobrinus, from the outermost to the innermost plaque. Ten consenting subjects were asked to form plaque by abstaining from tooth brushing over 3 days within in situ plaque-generating devices, which were placed on the upper molars. The plaque formed in the devices was separated into 8 - 10 layered fractions ( 100 mum thick). Genomic DNA was extracted from each plaque fraction by a commercial DNA purification kit and used for the amplification of the 16S ribosomal RNA gene sequences by polymerase chain reaction (PCR) with universal primers. The products were then amplified by PCR with S. mutans- or S. sobrinus-specific nested primers. The final products were separated on agarose gels, stained and photographed to confirm the existence of S. mutans and S. sobrinus. The results showed that S. mutans was detected in the plaque obtained from all of the 10 subjects and S. sobrinus in the plaque of 7 subjects. However, the distribution patterns of fractions positive for S. mutans and S. sobrinus varied among the subjects, with a tendency for frequent detection of both species in the outer to middle layers of dental plaque. There were no plaque fractions in which only S. sobrinus was found. This method could be useful to map the distribution of cariogenic microorganisms and to estimate the bacterial ecology for oral biofilm. Copyright (C) 2004 S. Karger AG, Basel.

DOI： 10.1159/000079626

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

Mutans streptococci are frequently isolated from dental plaque and carious lesions. These bacteria have been identified by conventional methods such as biochemical and serologic tests followed by the isolation of colonies on the mitis-salivarius agar, which are sometimes inconsistent. Recently, species-specific polymerase chain reaction (PCR) has been reported to rapidly identify Streptococcus mutans and Streptococcus sobrinus. However, in the case of identification and classification into several species, e.g. within the group of mutans streptococci consisting of seven species, the identification using species-specific PCR seems somewhat inefficient because of need for the development and preparation of specific primers for each species. Therefore, in this study we developed a simple method using restriction fragment length polymorphism analysis of PCR-amplified 16S ribosomal RNA genes (16S rRNA genes PCR-RFLP) for the identification of seven different species included in the group of mutans streptococci. We amplified 16S rRNA gene sequences from genomic DNA samples by PCR using universal primers and digested the PCR products with the restriction endonucleases, HpaII and HaeIII. HpaII produced six RFLP patterns for eight reference strains, since the patterns for S. sobrinus, Streptococcus downei and Streptococcus ferus were similar. RFLP patterns produced with HaeIII could separate these three species. Furthermore, RFLP patterns predicted from the 16S rRNA gene sequences in the GenBank database agreed with the actual RFLP patterns produced in the present study. The 16S rRNA sequence comparisons can be used to identify oral mutans streptococci; however, small laboratories. Therefore, 16S rRNA genes PCR-RFLP, using HpaII and HaeIII could be an alternative method for the identification of mutans streptococci, and may be applicable for large-scale studies on the cariogenicity of mutans streptococci.

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：BLACKWELL PUBLISHING LTD  

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.
Methods and Results: A universal set of PCR primers for bacterial 16S rRNA gene was introduced for the first PCR, and then two sets of primers specific for the 16S rRNA gene sequences of either Strep. mutans or Strep. sobrinus were used for the second PCR. Eighteen plaque samples were analyzed, and a nested PCR was shown to be more sensitive for detecting Strep. mutans and Strep. sobrinus than direct PCR.
Conclusions, Significance and Impact of the Study: The 16S rRNA gene-based nested PCR method is a rapid and sensitive method for the detection of mutans streptococci, and may also be suitable for carrying out large-scale studies on the cariogenicity of mutans streptococci.

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Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Fusobacterium nucleation, which can frequently be isolated from periodontal pockets, preferentially utilize proteins and peptides as growth substrates. In this study, we determined the size of peptide that is preferentially utilized as a source of energy and material for cell growth by P. gingivalis, P. intermedia, P. nigrescens and F nucleation using various sizes of poly amino acids consisting of two to approximately 100 molecules of aspartate or glutamate. Resting cells of P. gingivalis, P. intermedia and P. nigrescens utilized aspartylaspartate, while cells of P. gingivalis and F. nucleation utilized glutamylglutamate. The addition of aspartylaspartate to the culture medium increased the growth of P. gingivalis, P. intermedia and P. nigrescens, while the addition of glutamylglutamate promoted the growth of P. gingivalis and F nucleatum. These results clearly indicate that dipeptides such as aspartylaspartate and glutamylglutamate can be utilized as growth substrates for P. gingivalis, P. intermedia, P. nigrescens and F. nucleatum.

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Although Porphyromonas gingivalis is known to utilize peptides preferentially, instead of free amino acids, as the source of energy and cell material, there is only limited information on what sizes and kinds of peptide this bacterium preferentially utilizes. In this study, therefore, we tested aspartate or glutamate monopolymers consisting of from 2 to 100 amino acids as metabolic substrates for P. gingivalis. The washed cells of P. gingivalis consumed aspartylaspartate and glutamylglutamate, and produced large amounts of ammonia and organic acids such as propionate and butyrate, while the cells formed only small amounts of end-products from aspartate, glutamate, and other peptides longer than a dipeptide. P. gingivalis also metabolized valylvaline and leucylleucine and produced isobutyrate and isovalerate, respectively, only in the presence of aspartylaspartate or glutamylglutamate. This suggests a metabolic linkage between these dipeptides. These results clearly indicate that P. gingivalis utilizes dipeptides preferentially as its metabolic substrates.

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A novel group of oral spirochaetes that were specifically labeled by H9-2, a monoclonal antibody against the 37-kDa endoflagellar sheath protein of Treponema pallidum, have earlier been demonstrated in periodontitis-associated plaque samples; two cultivable oral spirochaetes T. vincentii and T. medium displayed cross-reactivity with that monoclonal, but the molecular basis for this cross-reactivity is not yet defined. Here, the flaA-1 genes which encode the 37-kDa sheath protein from T. vincentii and the oral spirochaetes present in samples from advanced periodontitis were examined. A 856 bp fragment of the flaA-1 gene was amplified in T. pallidum and T. vincentii. The same-size polymerase chain reaction fragments were also amplified in two clinical samples from patients with advanced periodontitis but not from samples of healthy plaque. The sequences of the flaA-1 genes of the oral spirochaetes detected in human periodontal plaque were closely similar to those of T. pallidum and T. vincentii, but neither of these two organisms could be detected in these samples using rRNA-specific primers. The identity of the flaA-1 positive spirochaetes associated with periodontitis remains to be determined. (C) 2000 Elsevier Science Ltd. All rights reserved.

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Metabolic pathways involved in the formation of cytotoxic end products by Porphyromonas gingivalis were studied. The washed cells of P. gingivalis ATCC 33277 utilized peptides hut not single amino acids. Since glutamate and aspartate moieties in the peptides were consumed most intensively, a dipeptide of glutamate or aspartate was then tested as a metabolic substrate of P. gingivalis. P. gingivalis cells metabolized glutamylglutamate to butyrate, propionate, acetate, and ammonia, and they metabolized aspartylaspartate to butyrate, succinate, acetate, and ammonia. Based on the detection of metabolic enzymes in the cell extracts and stoichiometric calculations (carbon recovery and oxidation/reduction ratio) during dipeptide degradation, the following metabolic pathways were proposed. Incorporated glutamylglutamate and aspartylaspartate are hydrolyzed to glutamate and aspartate, respectively, by dipeptidase. Glutamate is deaminated and oxidized to succinyl-coenzyme A (CoA) by glutamate dehydrogenase and 2-oxoglutarate oxidoreductase, Aspartate is deaminated into fumarate by aspartate ammonia-lyase and then reduced to succinyl-CoA by fumarate reductase and acyl-CoA:acetate CoA-transferase or oxidized to acetyl-CoA by a sequential reaction of fumarase, malate dehydrogenase, oxaloacetate decarboxylase, and pyruvate oxidoreductase. The succinyl-CoA is reduced to butyryl-CoA by a series of enzymes, including succinate-semialdehyde dehydrogenase, 4-hydroxybutyrate dehydrogenase, and butyryl-CoA oxidoreductase. A part of succinyl-CoA could be converted to propionyl-CoA through the reactions initiated by methylmalonyl-CoA mutase, The butyryl- and propionyl-CoAs thus formed could then be converted into acetyl-CoA by acyl-CoA:acetate CoA-transferase with the formation of corresponding cytotoxic end products, butyrate and propionate. The formed acetyl-CoA could then be metabolized further to acetate.

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Although oral treponemes are among the most frequently found bacteria in periodontal pockets, identification of these organisms can be difficult. In this study, restriction fragment-length polymorphism (RFLP) analysis of polymerase chain reaction (PCR)-amplified 16S ribosomal RNA genes (16S rRNA gene PCR-RFLP) was used to generate restriction profiles of reference strains of oral treponemes including Treponema denticola, Treponema socranskii, Treponema vincentii, Treponema pectinovorum and Treponema medium as well as for Treponema phagedenis and Treponema pallidum and five treponeme strains isolated from human periodontal pockets. Before RFLP analysis, the 16S rRNA gene sequences were obtained from the GenBank database, and the analysis of the theoretical banding patterns for HpaII suggested good species discrimination. 16S rRNA gene sequences were amplified from isolated genomic DNA samples by PCR with spirochete-specific primers. The PCR products were then purified and characterized by single digestion with restriction endonuclease HpaII, and this allowed discrimination between the respective reference strains. Five clinical isolates, four T. denticola and one T. socranskii, were assigned on the basis of their restriction profiles by digestion with HpaII. 16S rRNA gene PCR-RFLP using HpaII is a rapid and reliable method for differentiation of cultivable oral treponemes.

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16S I DNA 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., Mim, HaeIII, CfoI, or HpaII. Among them, Mn/I 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 Mn/I. Thus, 16S rDNA PCR-RFLP, using Mn/I, is a rapid and reliable method for the differentiation of oral Actinomyces spp. (C) 1998 Elsevier Science Ltd. All rights reserved.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER SOC MICROBIOLOGY  

We report the design and evaluation of PCR primers 63f and 1387r for amplification of 16S rRNA genes from bacteria. Their specificity and efficacy were tested systematically with a variety of bacterial species and environmental samples. They were found to be more useful for 16S rRNA gene amplification in ecological and systematic studies than PCR amplimers that are currently more generally used.

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：MUNKSGAARD INT PUBL LTD  

Restriction fragment-length polymorphism (RFLP) analysis of 16S rDNA amplified by polymerase chain reaction was used to generate restriction profiles of the type strains of oral asaccharolytic Eubacterium species, that is, Eubacterium brachy, Eubacterium exiguum, Eubacterium lentum, Eubacterium minutum, Eubacterium nodatum, Eubacterium saphenum, Eubacterium timidum and 33 asaccharolytic Eubacterium strains isolated from oral sites. The 16S rRNA gene sequences from isolated genomic DNA samples were amplified by polymerase chain reaction (PCR). PCR products were purified and characterized by single digestions with 7 restriction endonucleases. Among the 7 endonucleases, HpaII was found to discriminate the respective reference strains. Twenty-three isolates, out of 33, were assigned to one of the reference species, on the basis of their restriction profiles by digestion with HpaII. The remaining 10 isolates could not be assigned to any of the established species and constituted 4 distinct groups, each of which may be a new species.

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：MUNKSGAARD INT PUBL LTD  

Veillonella atypica, Veillonella dispar and Veillonella parvula cannot be reliably distinguished by conventional phenotypic tests, including the API ZYM test. In this study, restricted fragment-length polymorphism (RFLP) analysis of 16S rDNA amplified by polymerase chain reaction (PCR) was used to generate restriction profiles of the type strains of V. atypica, V. dispar and V. parvula and 20 Veillonella strains isolated from oral sites. 16S rRNA gene sequences from isolated genomic DNA samples were amplified by PCR. PCR products were purl fled and characterized by single digestion with 13 restriction endonucleases. Among them, MnlI was found to discriminate the respective reference strains, and the clinical isolates were assigned to one of the three species on the basis of their restriction profiles by digestion with MnlI. Thus, RFLP analysis of PCR-amplified 16S rDNA, using MnlI, is a rapid and reliable method for the differentiation of V. atypica, V. dispar and V. parvula.

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER SOC MICROBIOLOGY  

Restriction fragment length pollymorphism 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. atypitca 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.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Japanese Association for Oral Biology  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Microbiology Society  

Eubacterium exiguum sp. nov. is the name proposed for organisms formerly described as Eubacterium group S strains and similar bacteria isolated from various types of oral lesions. This new species was established on the basis of the results of DNA-DNA hybridization experiments and DNA base composition determinations (G+C contents, 60 to 64 mol%). The results of an API ZYM analysis, Western blotting (immunoblotting) reactions, and phenotypic tests are also given. The type strain of E. exiguum is strain S-7.

DOI： 10.1099/00207713-46-4-1120

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The purpose of this study was to clarify the antibacterial efficacy of mixed antibacterial drugs on bacteria of carious and endodontic lesions of human deciduous teeth in vitro. The antibacterial drugs used in this study were mixtures of ciprofloxacin, metronidazole, plus a third antibiotic: amoxicillin, cefaclor, cefroxadine, fosfomycin or rokitamycin. Samples taken from carious dentin (17 cases) and infected pulpal tissues (14 cases) were cultured on control plates and plates containing the mixed drugs. No bacteria were recovered in the presence of any combination of the mixture of the drugs (100 mug each/ml), and the bacterial growth occurred on control plates (10(1) to 10(7) Colony-forming units), indicating that the mixed drugs inhibit the growth of bacteria in the samples. When carious and endodontic lesions on split surfaces of freshly extracted teeth were covered overnight with alpha-tricalcium phosphate cement containing a mixture of ciprofloxacin, metronidazole and cefaclor (1% each; 5 cases), no bacteria were recovered from the lesions. No bacteria were recovered from carious and endodontic lesions when these lesions were immersed in a solution of the mixture (200 mug each/ml; 5 cases). These findings indicate that carious and endodontic lesions can be sterilized by the mixed drugs in situ.

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Informa Healthcare  

The purpose of this study was to investigate the bacterial composition of necrotic pulps of human deciduous teeth by sampling the split surfaces of freshly extracted teeth and culturing the bacteria present with good anaerobic isolation techniques. Significantly more bacteria were recovered after the incubation in an anaerobic chamber than after aerobic incubation in air with 30 per cent CO2. Of 276 bacterial isolates, 251 (91 per cent) were obligate anaerobes. These findings suggest that the environment of necrotic pulps in human deciduous teeth is anaerobic and thus favours the growth of anaerobes. Among the 251 obligate anaerobes isolated, strains belonging to the genera Peptostreptococcus (25 per cent), Propionibacterium (19 per cent), Eubacterium (17 per cent) and Fusobacterium (13 per cent) were major parts of the bacterial flora of the lesions of human deciduous teeth. Bifidobacterium (2 per cent), Lactobacillus (1 per cent), Actinomyees (1 per cent) and Veillonella (0.7 per cent) were minor parts of the flora. The microflora of necrotic pulps of human deciduous teeth is in some respects similar to that reported for the deep layers of dentinal lesions of adults. ©1993 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.

DOI： 10.3109/08910609309141335

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Informa Healthcare  

The aim of this study was to clarify the antibacterial efficacy of a mixture of ciprofloxacin, metronidazole and minocycline, with and without the addition of rifampicin (100 μg each/ml), against oral bacteria of children. More than 101 bacteria occurred in samples taken from carious (32 cases) and endodontic (28 cases) lesions, and from periodontal pockets (seven cases). However, none were recovered in vitro in the presence of the mixture. The mixture was also effective against the bacteria of dental plaque formed on deciduous teeth (16 cases). When carious and endodontic lesions on split surfaces of freshly extracted teeth were covered overnight with αtricalcium phosphate (TCP) cement containing the mixed drugs (1 per cent each
seven cases) or immersed in a solution of the mixed drugs (200 μg each/ml
two cases), no bacteria were recovered from the lesions, indicating that the mixture sterilised the lesions in situ. Topical application of the mixture of the drugs to carious and endodontic lesions, dissolved in water or mixed into dental materials, may be useful in denial treatment for children. ©1992 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.

DOI： 10.3109/08910609209141583

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The aim of this study was to quantify cariogenic bacteria, Streptococcus mutans, in supragingival plaque biofilm microflora on caries lesions of children. After informed consent was obtained from each subject, supragingival plaque on caries lesion was obtained from 10 children (3-10 years). Healthy supragingival plaque was also obtained from the same subject, as a control. Total bacteria and S. mutans were quantified by real-time PCR using universal and S. mutans-specific primers based on 16S ribosomal RNA genes, respectively, and the proportion of S. mutans in each sample was calculated. Total bacterial DNA levels in carious plaque and healthy plaque were found to be similar, though the proportion of S. mutans in carious plaque was higher than that in healthy plaque. The findings of this study suggest that amount of cariogenic bacteria in supragingival plaque biofilm may be associated with the status of dental caries.

DOI： 10.1007/978-4-431-99644-6_75

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A method for determining the profiles of biomass volume and glucan within plaque was developed using depth-specific analysis of plaque. The profiles of biomass volume and glucan demonstrated a tendency to be higher in the plaque exposed to sucrose, suggesting that an evaluation of these two plaque indices would be important to see the cariogenicity in the diet.

DOI： 10.1007/978-4-431-99644-6_61

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The frequency of periodontal diseases appears to increase with age. Porphyromonas gingivalis is widely regarded as major periodontal pathogens. This study aimed to quantify P gingivalis in subgingival plaque biofilm of elderly subjects by real-time polymerase chain reaction (PCR). Subgingival plaque was obtained from 198 periodontally healthy (mean age, 70.3 years) and 176 subjects with periodontitis (70.6 years). Quantification of total bacteria and P gingivalis was performed by real-time PCR using universal and P gingivalis-specific primers based on 16S rRNA genes, respectively. Both the detection frequency and mean proportion of P gingivalis were significantly higher in subjects with periodontitis than in periodontally healthy subjects (p &lt; 0.0001). Nevertheless, P gingivalis was detected frequently both from subjects with periodontitis and periodontally healthy subjects. These results suggest that P. gingivalis is widely distributed in subgingival plaque biofilm of elderly subjects.

DOI： 10.1007/978-4-431-99644-6_62

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：SPRINGER-VERLAG TOKYO  

Postoperative pneumonia (PP) occurs in elderly subjects whose cough reflex (CR) is impaired postoperatively. This finding indicates that silent aspiration of oral bacteria may be involved in PP. This study aimed to clarify the involvement of CR impairment and silent aspiration of oral bacteria in PP. Intraoperative bronchial aspirates (BA) from 22 subjects aged over 60 years undergoing lung resection were cultured anaerobically on blood agar plates. The CR of all subjects was measured on 1st postoperative day. Eleven subjects showed impaired CR, whereas 11 showed normal. The bacterial amounts in intraoperative BA of subjects with postoperatively impaired CR were found to be higher than those with normal CR. PP occurred in 2 out of 11 subjects with impaired CR but not in 11 subjects with normal CR. These results suggest that impairment of the CR and silent aspiration of oral bacteria may be associated with the development of PP in elderly subjects.

DOI： 10.1007/978-4-431-99644-6_76

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This study aimed to clarify (1) what kind of symptoms the elderly were aware of, (2) the relationship between those symptoms and oral diseases, and (3) the relationship between those symptoms and oral health behaviors. Subjects consisted of 459 individuals 60 years and over, who were asked about subjective symptoms and oral health behaviors, and given an oral health examination. Findings were: (1) even though most subjects (75.2%) had the subjective symptoms, 55.7% of them did not think of them as health problems, (2) logistic regression analysis revealed that those who had subjective symptoms were at higher risk to have decayed teeth, periodontitis, and missing teeth (p &lt; 0.01-0.05), and (3) the elderly who had oral complaints or the subjective symptoms used an interdental brush or a dental floss much more often than those who did not (p &lt; 0.05). However, the elderly who had the oral complaint showed negative responses towards the visiting dentists (p &lt; 0.05).

DOI： 10.1007/978-4-431-99644-6_102

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This study aimed to profile plaque microflora on root caries lesions and to examine the protein-degrading activity of isolated bacteria. Plaque samples on root caries lesions (R) and from healthy supragingival sites (S) of six subjects were collected and cultured anaerobically on blood agar plates. The isolated bacteria were identified by 16S rRNA sequencing and examined for their protein-degrading activity, using the skim-milk plates, and for their acidogenicity, using the FAB broth. Propionibacterium, Actinomyces, Streptococcus, Lactobacillus, and Bifidobacterium were predominant in R. The skim-milk plates distinguished between protein-degrading and protein-coagulating bacteria, which comprised 7 and 33% of microflora in R, respectively. These results suggest that protein-coagulating bacteria demineralize hydroxyapatite and denature proteins of root dentin, and that protein-degrading bacteria may degrade the denatured proteins.

DOI： 10.1007/978-4-431-99644-6_63

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This study aimed to profile plaque microflora on first molars with orthodontic bands (Ba), brackets (Br), and without appliances (C). The mean bacterial numbers of plaque (logCFUs/mg) on the molars with Ba, Br, and C were 6.6, 6.7, and 6.9, respectively. Actinomyces, Streptococcus, and Veillonella were predominant in Br and C, while the proportions of Actinomyces and Veillonella were low in Ba. Periodontitis-associated bacteria including Eubacterium, Fusobacterium, Porphyromonas, and Prevotella were isolated in Br, but virtually not detected in Ba and C, suggesting that supragingival plaque biofilm of teeth with Br carries bacteria related to periodontitis. These findings may provide a helpful suggestion for self-care and regular professional plaque control in clinical orthodontics.

DOI： 10.1007/978-4-431-99644-6_65

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The profile of bacterial density within plaque analyzed by real-time PCR indicated that SnF2 gel treatment promoted a larger bacterial habitat, particularly in deeper regions. On the other hand, the treatment resulted in unpredictable changes in S. mutans' habitat. © 2005 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.ics.2005.07.010

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The aim of this study was to clarify the microflora in crevices around titanium orthodontic anchor plates using anaerobic culture method and molecular biological technique for bacterial identification, and to compare microbial composition between crevices around anchor plates and gingival crevices. Samples from crevices around titanium anchor plates and healthy gingival crevices were cultured anaerobically, and the isolated bacteria were identified by 16S rRNA sequencing. The bacterial density of the crevices was lower than that of healthy gingival crevices. In healthy plate crevices of seven subjects, out of 184 strains isolated as predominant bacteria, 108 were anaerobic bacteria, while 73 were facultative bacteria. In inflamed plate crevices of three subjects, out of 133 strains isolated as predominant bacteria, 110 were anaerobic bacteria. On the other hand, in healthy gingival crevices of seven subjects, out of 146 strains isolated as predominant bacteria, 98 were facultative bacteria, while 45 were anaerobic bacteria. These results suggest that the environment in crevices around titanium orthodontic anchor plates is anaerobic and supportive of the growth of anaerobes, which may trigger inflammation of tissue around the plates. © 2005 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.ics.2005.06.061

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The aim of this study was to compare the detection frequencies of 25 bacterial species in subgingival with supragingival plaque of periodontitis subjects and periodontally healthy subjects. Two species (Mogibacterium timidum and Porphyromonasgingivalis) were detected significantly more frequently in subgingival plaque of periodontitis subjects than of healthy subjects, suggesting that these two species are closely related to periodontitis. There were no significant differences in the detection frequencies of the 25 bacteria between subgingival and supragingival plaque, suggesting that the bacterial flora of supragingival plaque reflects that of subgingival plaque. © 2005 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.ics.2005.06.069

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Relationship between bacterial flora and H2S levels in breath air was investigated in this study. Oral malodor levels in 10 subjects (age: 21-56 years) were assessed with gas chromatography, Breathtron® and organoleptic score. Based on these assessments, subjects were divided into 2 groups
oral malodorous and nonodorous groups. Tongue coating were sampled and spread onto Fastidious Anaerobe Agar plates containing 0.05% cysteine, 0.12% glutathione and 0.02% lead acetate, and were then incubated anaerobically at 37 °C for 2 weeks. Bacteria forming black or gray colonies were designated as H2S-producing phenotypes, and were identified by 16S rRNA gene sequencing. The numbers of total bacteria (p &lt
0.005) and H2S-producing bacteria (p &lt
0.05) in the oral malodorous group were significantly larger than those in the nonodorous group. Veillonella, Actinomyces and Prevotella species were predominant in the H2S-producing bacteria both in the oral malodorous and nonodorous groups. These results suggest that an increase in the number of H2S-producing bacteria in the tongue biofilm is responsible for oral malodor. © 2005 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.ics.2005.06.068

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The aims of this study were to determine the bacterial density and to examine the presence of 20 bacteria in fluids of postoperative maxillary cysts (POMC). POMC fluids (4 purulent, 2 mucous and 4 serous) were sampled from 10 subjects (aged 43 to 77). Bacterial quantification and detection were performed by real-time PCR and nested PCR based on bacterial 16S rRNA genes, respectively. Bacterial DNA was detected in all samples and the average concentrations of bacterial DNA were 5.9 (purulent), 0.5 (mucous) and 0.7 (serous) ng/mg of sample. Twelve bacterial species were detected in the purulent fluids, while 2 and 5 species were detected in the mucous and serous fluids, respectively. Purulent fluids contained numerous bacteria of various types, suggesting that oral bacteria cause symptoms such as pain in POMC with purulent fluids. Mucous and serous fluids also contained bacteria, although their numbers were small. © 2005 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.ics.2005.06.062

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高齢者の歯髄の免疫防御能を明らかにする基礎的データを提供するために,ネズミ(ラット)の歯を用いて歯髄を口腔内に開放して感染を起こした後の抗菌性薬剤に対する歯髄反応を,初期免疫防御反応に重要な役割を果たす抗原提示細胞のマーカーを用いて免疫細胞化学的に明らかにするとともに,幼若ラットと高齢ラットで歯牙切削に対する歯髄の反応性の違いについて検索した.抗菌性薬剤添加により感染歯髄が再生し,歯髄・象牙境への抗原提示細胞の集積が歯髄再生の必須の過程であることが明らかとなった.さらに,高齢ラットにおいて歯髄免疫防御・修復機能が保持されている事が明らかとなったが,窩洞形成後に象牙芽細胞の反応性の違いが観察され,加齢により象牙芽細胞突起の状態が変化する可能性が示唆された

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Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)   Publisher：日本歯科評論社  

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Venue：Portland, Oregon, USA   Country：United States  

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Language：Japanese   Presentation type：Oral presentation (general)  

Venue：新潟  

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《最優秀賞》受賞

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Language：Japanese   Presentation type：Oral presentation (general)  

Venue：仙台  

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Language：Japanese   Presentation type：Oral presentation (general)  

Venue：仙台  

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Presentation type：Symposium, workshop panel (public)  

Venue：福岡  

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Presentation type：Symposium, workshop panel (public)  

Venue：福岡  

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Language：English   Presentation type：Symposium, workshop panel (public)  

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Language：Japanese   Presentation type：Oral presentation (general)  

Venue：オンライン  

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Language：Japanese   Presentation type：Oral presentation (general)  

Venue：大阪   Country：Japan  

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Venue：オンライン  

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Presentation type：Symposium, workshop panel (public)  

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Language：Japanese   Presentation type：Oral presentation (general)  

Venue：仙台  

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Language：Japanese   Presentation type：Oral presentation (general)  

Venue：仙台  

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Presentation type：Symposium, workshop panel (public)  

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Presentation type：Symposium, workshop panel (public)  

Venue：仙台  

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Presentation type：Symposium, workshop panel (public)  

Venue：仙台  

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Presentation type：Symposium, workshop panel (public)  

Venue：札幌  

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Language：Japanese   Presentation type：Symposium, workshop panel (nominated)  

Venue：仙台  

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Language：English   Presentation type：Symposium, workshop panel (nominated)  

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Language：English   Presentation type：Symposium, workshop panel (public)  

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Language：English   Presentation type：Symposium, workshop panel (public)  

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Presentation type：Symposium, workshop panel (nominated)  

Country：Thailand  

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Language：English   Presentation type：Symposium, workshop panel (nominated)  

Venue：Beijing   Country：China  

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Language：Japanese   Presentation type：Symposium, workshop panel (nominated)  

Venue：仙台  

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Language：Japanese   Presentation type：Symposium, workshop panel (nominated)  

Venue：長崎  

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Language：Japanese   Presentation type：Symposium, workshop panel (nominated)  

Venue：仙台  

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Language：Japanese   Presentation type：Symposium, workshop panel (public)  

Venue：森岡  

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Country：Japan

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Country：Japan

一時金30万円

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Country：Japan

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Country：Japan

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Country：Japan

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Country：Japan

10万円

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Country：Japan

35万円

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Award type：Award from publisher, newspaper, foundation, etc.  Country：Japan

月額5万円×1年間

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Country：Japan

入学金：実費、授業料：実費、書籍および下宿補助（月額6万円）

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Country：Japan

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Award type：Award from publisher, newspaper, foundation, etc.  Country：Japan

5万円

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Award type：Award from Japanese society, conference, symposium, etc.  Country：Japan

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Award type：Award from Japanese society, conference, symposium, etc.  Country：Japan

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Country：Japan

採択額 10万円

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給付額；35万円

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給付額；144万円

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Award type：Award from Japanese society, conference, symposium, etc.  Country：Japan

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Award type：Award from Japanese society, conference, symposium, etc. 

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給付額；144万円

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給付額；198万円

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211万2千円

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給付額；35万円

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給付額；30万円

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Award type：Award from Japanese society, conference, symposium, etc.  Country：Japan

副賞；2万円

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Award type：Award from Japanese society, conference, symposium, etc. 

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給付額；20万円

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給付額；120万円

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