• Title/Summary/Keyword: Prevotella intermedia

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The Frequency of Detecting Prevotella intermedia and Prevotella nigrescens in Korean Adult Periodontitis Patients (한국인 치주 감염 환자에서의 Prevotella intermedia와 Prevotella nigrescens의 발현빈도)

  • Peck, Seung-Yup;Ku, Young;Rhyu, In-Cheol;Hahm, Byung-Do;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.419-429
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    • 2000
  • Prevotella intermedia has been implicated as a potent pathogen in many kinds of periodontal, pulpal and periapical diseases. However, it has been isolated from periodontally healthy adults and from edentulous children as well. The intraspecies heterogeneity of Prevotella intermedia has been demonstrated in early studies and finally Shah & Gharbia confirmed the existence of 2 DNA homology groups and proposed dividing Prevotella intermedia into 2 species, Prevotella intermedia and Prevotella nigrescens. This study was designed to examine the frequency of Prevotella intermedia and Prevotella nigrescens in diseased periodontal pockets and healthy gingival sulcus of Korean people by PCR based on 16s ribosomal DNA sequence. One hundred adults who had adult periodontitis but not taken any periodontal treatment or antibiotics during previous 6 months and 50 adults who had healthy periodontal tissue were selected for this study. The sulcular fluid was collected into VMGA by sterilized paper point and diluted to 1,000 times in anaerobic chamber. $100{\mu}{\ell}$ of sample was cultured in $37^{\circ}C$ for 10 days. Among the bacterial colonies, BPB were selected and cultured in BHI broth and then Prevotella intermedia was identified through Gram staining and biochemical test. Identified Prevotella intermedia was cultured again and centrifuged. DNA was extracted from the pellet using several reagents. PCR was performed by previously designed primer. The results were followed. 1. BPB were isolated from 39 of 100 samples of diseased periodontal pockets(39%). 2. Prevotella intermedia was identified from 24 of 39 BPB samples. 3. Among 24 Prevotella intermedia, 21 were confirmed as Prevotella inter - media(87.5) and 2 were confirmed as Prevotella nigrescens(8.33%). 4. BPB were isolated from 9 of 50 samples of periodontally healthy patients. Among them only two were identified as Prevotella intermedia, that is, one was confirmed as Prevotella intermedia and the other was Prevotella nigrescens.

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EFFECT OF POLYPHOSPHATE ON THE GROWTH OF ORAL BACTERIUM, PREVOTELLA INTERMEDIA (구강세균 Prevotella intermedia의 성장에 따른 polyphosphate의 영향에 관한 연구)

  • Kong, Hee-Joung;Choi, Ho-Young;Min, Byung-Soon;Part, Sang-Jin;Lee, Jin-Yong;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.23 no.2
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    • pp.550-560
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    • 1998
  • Prevotella intermedia has been known as one of the important bacterial species involved in the endodontic infections and various periodontal diseases. Polyphosphate has been widely used to prevent decomposition of food and known to have an inhibitory effect on the growth of gram positive bacteria. The purpose of this study was to evaluate the effect of poly phosphate on the growth of Prevotella intermedia, a gram negative bacterium. Prevotella intermedia G8GK3(ATCC 49046) was grown in the presence of polyphosphates with different chain lengths. Inhibitory effect of each polyphosphate, which was added at the beginning or at the early exponential growth phase of Prevotella intermedia, was determined by measuring optical density of the bacterial cells at 540nm, viable cells and lysis of Prevotella intermedia. The results from this study were as follows : 1. Poly phosphate inhibited the growth of Prevotella intermedia. 2. The minimum inhibitory concentration(MIC) of poly phosphate appeared to be 0.05%. 3. Polyphosphates with chain lengths of 5 and 65 demonstrated the greatest inhibitory effect on the growth of Prevotella intermedia. 4. Polyphosphate was bactericidal to Prevotella intermedia, demonstrating the growth inhibition of the bacterium. 5. Polyphosphate induced lysis of Prevotella intermedia. The overall results suggest that polyphosphate has a bactericidal effect on Prevotella intermedia, causing the lysis of the bacterium.

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Interleukin-8 production and interleukin-8 mRNA expression induced by lipopolysaccharides from Prevotella intermedia and Prevotella nigrescens in monocyte-derived macrophages (Prevotella intermedia 및 Prevotella nigrescens의 지질다당질이 대식 세포에서의 Interleukin-8 생성에 미치는 영향)

  • Kim, Sung-Jo
    • Journal of Periodontal and Implant Science
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    • v.39 no.2
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    • pp.177-184
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    • 2009
  • Purpose: Interleukin-8 (IL-8) is an important mediator of immune and inflammatory reactions and is produced by a variety of different cell types. This study was undertaken to investigate the effects of lipopolysaccharides (LPSs) from Prevotella intermedia and Prevotella nigrescens, the major causes of inflammatory periodontal disease, on the production of IL-8 and the expression of IL-8 mRNA in differentiated THP-1 cells, a human monocytic cell line. Methods: LPSs from P. intermedia ATCC 25611 and P. nigrescens ATCC 33563 were prepared by the standard hot phenol-water method. THP-1 cells were incubated in the medium supplemented with phorbol myristate acetate to induce differentiation into macrophage-like cells. Results: We found that LPS preparations from P. intermedia and P. nigrescens can induce IL-8 mRNA expression and stimulate the release of IL-8 in differentiated THP-1 cells without additional stimuli. Conclusions: There are no previous reports of the ability of P. intermedia and P. nigrescens LPS to stimulate the release of IL-8, and the present study clearly shows, for the first time, that LPSs from P. intermedia and P. nigrescens fully induced IL-8 mRNA expression and IL-8 production in differentiated human monocytic cell line THP-1. The ability of P. intermedia and P. nigrescens LPS to promote the production of IL-8 may be important in the pathogenesis of inflammatory periodontal disease.

Chemical and Immunobiological Characterization of Lipopolysaccharides from Prevotella intermedia and Prevotella nigrescens (Prevotella intermedia와 Prevotella nigrescens의 세균내독소에 대한 연구;화학적 분석 및 면역생물학적 활성 평가)

  • Kim, Sung-Jo
    • Journal of Periodontal and Implant Science
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    • v.34 no.2
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    • pp.461-474
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    • 2004
  • The purpose of this study was to assess some biological activities of lipopolysaccharides (LPSs) from P. intermedia and P. nigrescens. LPS was prepared by the standard hot phenol-water method. NO production was assayed by measuring the accumulation of nitrite in culture supernatants. $TNF-{\alpha}$ production was determined by enzyme-linked immunosorbent assay. Western blot analysis of iNOS and analysis of reverse transcription (RT)-PCR products were carried out. LPS from P. intermedia demonstrated higher KDO content than those from two stains of P. nigrescens. LPSs from P. intermedia and P. nigrescens were mitogenic for spleen cells of BALB/C mouse. The present study clearly shows that LPSs from P. intermedia and P. nigrescens fully induced iNOS expression and NO production in RAW264.7 cells in the absence of other stimuli. Moreover, LPSs from P. intermedia and P. nigrescens clearly induced $TNF-{\alpha}$ production in RAW264.7 cells. The biological activities of LPS from P. intermedia was found to be comparable to those of P. nigrescens LPS. The ability of LPSs from P. intermedia and P. nigrescens to promote the production of NO and $TNF-{\alpha}$ may be important in the pathogenesis of inflammatory periodontal disease.

Isolation and Partial Characterization of Hemin-binding Cell Envelope Proteins from Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens (Porphyromonas gingivalis, Prevotella intermedia, 그리고 Prevotella nigrescens에서의 hemin 결합 단백질에 대한 연구)

  • Kim, Sung-Jo
    • Journal of Periodontal and Implant Science
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    • v.36 no.1
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    • pp.155-165
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    • 2006
  • The results of this study confirm that the availability of hemin influences the expression of selected membrane proteins of Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. A 30 kDa (heated 24 kDa) hemin-binding protein whose expression is hemin regulated was identified and purified in P. gingivalis. A strong hemin-binding function was found by LDS-PAGE and TMBZ staining when P. gingivalis cells were grown under hemin-limited conditions. A 50 kDa cell envelope associated protein, whose expression is hemin regulated, is considered to be a putative hemin binding protein from P. intermedia and P. nigrescens, respectively. N-terminal amino acid sequence analysis of CNBr-digested 24 kDa hemin binding protein from P. gingivalis revealed that this protein belongs to a new, so far undescribed hemin-binding class of proteins. N-terminal amino acid sequence of a 50 kDa putative hemin binding protein from P. intermedia was identical with Enolase from Streptococcus intermedia. Work is in progress to further characterize the molecular structure of these proteins.

Identification of Prevotella intermedia ATCC 25611 Using Pi29-L DNA Probe. (Pi29-L DNA 프로브를 이용한 Prevotella intermedia ATCC 25611의 동정)

  • 국중기;백동헌
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.205-209
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    • 2003
  • Recently, we introduced a new method for rapid screening of bacterial species- or subspecies-specific DNA probes, named “inverted dot blot hybridization screening method”. We then applied this method to develop species- or strain- specific DNA probes for Prevotella intermedia and Prevotella nigrescens. In those studies, among 96 candidate DNA probes which were screened by the new method, 5 probes were confirmed as being putatively strain-specific : 3 probes for P. nigrescens 9336 (ATCC 33563), one for each p. intermedia ATCC 25611 and one for P. nigrescens G8-9K-3 (ATCC 49046). In the present study, we evaluated by Southern blot analysis a DNA probe Pi29-L, one of the 96 candidate probes described above, whether it is specific for the strain ATCC 25611 off. intermedia. Our data show that the probe Pi29-L is potentially P. intermedia ATCC 25611-specific, which can be useful for the detection and identification of the strain, particularly in maintenance of the strain.

Draft genome sequence of Prevotella intermedia KCOM 1107 isolated from a human subgingival dental plaque of gingivitis lesion (사람 치은염 병소의 치은연하치면세균막에서 분리된 Prevotella intermedia KCOM 1107의 유전체 염기서열 해독)

  • Park, Soon-Nang;Lim, Yun Kyong;Shin, Ja Young;Roh, Hanseong;Kook, Joong-Ki
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.222-224
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    • 2017
  • Prevotella intermedia is a Gram-negative, obligately anaerobic, nonsporeforming, and nonmotile rod. P. intermedia is associated with periodontitis, pregnancy gingivitis, acute necrotic ulcerative gingivitis, endodontic infection, and rheumatoid arthritis. P. intermedia KCOM 1107 (= ChDC KB29) was isolated from a human subgingival dental plaque of gingivitis lesion. Here, we present the draft genome sequence of P. intermedia KCOM 1107.

Strain-specific PCR Primers for the Detection of Prevotella intermedia ATCC 49046

  • Kim, Min-Jung;Min, Jeong-Bum;Lim, Sun-A;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.36 no.2
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    • pp.79-82
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    • 2011
  • The aim of this study was to develop Prevotella intermedia ATCC 49046-specific PCR primers designed based on the nucleotide sequence of a DNA probe Pig28. The strainspecificity of the PCR primers, Pig28-F1/Pig28-R1, was confirmed with 9 strains of P. intermedia and 25 strains (15 species) of Prevotella species. The detection limit of the PCR primers was 2 pg of the purified genomic DNA of P. intermedia ATCC 49046. These PCR primers were found to be useful for identifying P. intermedia ATCC 49046, particularly for determining the authenticity of the strain.

Isolation and Characterization of a Putative Heminbinding Protein from Prevotella intermedia (Prevotella intermedia에서의 hemin 결합 단백질의 순수분리 및 특성분석)

  • Kim, Sung-Jo;Chung, Hae-Young
    • Journal of Periodontal and Implant Science
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    • v.30 no.4
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    • pp.737-746
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    • 2000
  • 본 연구는 hemin이 치주질환 주요 병인균주 중의 하나인 Prevotella intermedia의 성장 및 세포막 단백질의 발현에 미치는 영향을 규명하고, hemin 결합에 관여하는 것으로 추정되는 단백질의 순수분리 및 특성 분석을 위해 수행 되었다. 본 연구의 결과에 의하면, hemin은 P.intermedia의 성장 및 세포막 단백질의 발현에 영향을 미쳐, hemin이 고갈된 조건에서 균주의 성장이 현저히 억제되었으며, 약 50 kDa의 세포막 단백질이 현저히 강화되어 발현되었다. 본 연구에서는 hemin 결합에 관여하는 것으로 추정되는 이 50 kDa의 세포막 단백질을 순수분리하였으며, N'-terminal 아미노산 분석을 수행한 결과 이 단백질은 Streptococcus inter - medius의 Enolase와 아미노산 서열 및 분자량이 일치하였다. 한편, 이 단백질에는 disulfidebond가 존재하지 않았다. 본 연구는 P.intermedia에서의 porphyrin 생리 및 hemin 획득기전을 밝히는데 있어 중요한 의의가 있으리라 사료된다.

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The Anti-inflammatory Effect of Green Tea Extract Against Prevotella intermedia (녹차추출물의 잇몸 질환 원인균에 대한 항염증 효능 연구)

  • Min, Dae-Jin;Yi, Sung-Won;Lee, Sung-Hoon;Kim, Seung-Seob;Kim, Chan-Ho;Lee, John-Hwan;Bae, Ji-Hyun;Kim, Han-Kon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.37 no.1
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    • pp.67-73
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    • 2011
  • Dental bacteria can cause gum diseases, i.e. gingivitis and periodontitis, by inducing inflammation in human gingiva. Therefore, the most effective way to prevent and treat gum diseases is the control of the inflammatory reactions induced by dental bacteria. Almost all present dental care products contain anti-bacterial agents to eliminate dental bacteria. However, recent studies report that even heat-killed dental bacteria can induce the inflammation responses in oral cells. Therefore, the method using anti-bacterial agents should be improved for better anti-inflammatory effect and the effective natural anti-inflammatory substances need to be found. In addition, the mechanisms of gingival inflammation should be elucidated. In this study, we tried to find out the mechanism of the gingival inflammation and effective natural anti-inflammatory substances with human gingival epithelial cells and Prevotella intermedia which is well known as a typical dental bacteria inducing gingivitis and periodontitis. In results, Prevotell intermedia initiated the gingival inflammation response by stimulating gingival epithelial cells to release an inflammatory cytokine, IL-8. Furthermore, the inflammation by Prevotella intermedia is related to COX-2, AP-1, and TNF-${\alpha}$ pathways. Green tea extract could effectively suppress the inflammatory responses induced by Prevotella intermedia. We find out the effective natural substance for the improvement of gum diseases by studying the mechanism of the gingival inflammation induced by dental bacteria.