Project/Area Number |
17390502
|
Research Category |
Grant-in-Aid for Scientific Research (B)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Pathobiological dentistry/Dental radiology
|
Research Institution | Okayama University |
Principal Investigator |
FUKUI Kazuhiro Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Professor (70034171)
|
Co-Investigator(Kenkyū-buntansha) |
KOKEGUCHI Susumu Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Associate Professor (10144776)
TANIMOTO Ichiro Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Assistant Professor (00280686)
TAKASHIBA Shogo Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Professor (50226768)
MAEDA Hiroshi Okayama University, Hospital, Lecturer (00274001)
KARIYAMA Reiko Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Assistant Professor (40112148)
玉木 直文 岡山大学, 大学院・医歯薬学総合研究科, 助手 (20335615)
|
Project Period (FY) |
2005 – 2007
|
Project Status |
Completed (Fiscal Year 2007)
|
Budget Amount *help |
¥16,260,000 (Direct Cost: ¥15,300,000、Indirect Cost: ¥960,000)
Fiscal Year 2007: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2006: ¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 2005: ¥5,400,000 (Direct Cost: ¥5,400,000)
|
Keywords | Bacteria / Metagenome / Biofilm / Infectious Disease / ゲノム |
Research Abstract |
Oral microflora has been investigated so far with cultivation-based techniques. Over 700 species of bacteria have been previously estimated in oral cavity but only about 50% of them have been cultivated. Any understanding of the oral environment requires knowledge of the entire bacterial community. The uncultivated and as-yet-uncharacterized bacterial species may participate in the etiology of oral diseases. To resolve this problem, we devised an approach by the metagenomic analysis based on the bacterial 16S ribosomal RNA gene (16S rDNA) . In this study, we determined the profiles, composition and changes of various oral microflora using culture-independent molecular methods, i.e., clone library method, polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) method and terminal restriction fragment length polymorphism analysis (T-RFLP) method. We also established the high-resolution, real-time and three-dimensional imaging system on the biofilm structure and deve
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lopment in the modified capillary flow-cell using confocal laser scanning microscopy with fluorescence visualization supporting by Professor Philip S. Stewart (Center for Biofilm Engineering at Montana State University). We analyzed the microbial profiles and composition of tonsilloliths, which are potential cause of oral malodor by 16S rDNAs based clone library method. The isolated partial 16S rDNA sequences (approximately 600bp) were analyzed. Anaerobic bacteria detected in tonsilloliths, belonged to the genera Fusobacterium, Eubacterium, Megasphaera, Porphyromonas, Prevotella, Selenomonas and Tannerella, which appear to be associated with production of volatile sulfur compounds. We further examined the effectiveness of professional toothbrushing on microflora changes in subgingival plaques by PCR-DGGE analysis. PCR-DGGE analysis revealed that professional toothbrushing resulted in a decrease in the number of periodontal pathogens and the dramatic change of microflora in subgingival plaques. We also revealed that Archaea was frequently isolated from deep periodontal pockets in Japanese patients with periodontitis and pathogenic and opportunistic bacterial species were frequently detected in the patients receiving bone marrow transplantation after chemotherapy. Less
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