• Journal of Periodontal and Implant Science
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• 제34권2호
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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.

• International Journal of Oral Biology
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• 제40권4호
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• pp.205-210
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• 2015

Prevotella intermedia-specific quantitative real-time PCR (qPCR) primers were previously designed based on the nucleotide sequences of RNA polymerase ${\beta}$-subunit gene (rpoB). However, the several clinical strains isolated from Korean populations are not detectable by the qPCR primers. The purpose of this study was to develop new P. intermedia-specific qPCR primers based on the rpoB. The specificity of the primers was determined by conventional PCR with 12 strains of P. intermedia and 52 strains (52 species) of non-P. intermedia bacteria. The sensitivity of primers was determined by qPCR with serial dilutions of the purified genomic DNAs (40 ng to 4 fg) of P. intermedia ATCC $25611^T$. The data indicated that only P. intermedia strains were detected by the P intermedia-specific qPCR primers (RTPiF2/RTPiR2); in addition, as little as 40 fg of P. intermedia genomic DNA could be detected. These results suggest that these qPCR primers are useful in detecting P. intermedia from the bacterial infectious lesions including dental plaque and oral tissue lesions.

• 미생물학회지
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• 제53권3호
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• pp.222-224
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• 2017

Prevotella intermedia는 그람 음성이면서, 절대 혐기성, 비스포아형성 및 운동성 간균이다. P. intermedia는 사람의 구강내 정상세균총의 하나이며, 치주질환, 임신성 치은염, 급성괴사성궤사성 치은염, 치근관 감염, 및 류마티성 관절염과 관련이 있다. P. intermedia KCOM 1107 (= ChDC KB29) 균주가 사람 치은염 병소 치은연하 치면세균막에서 분리되었다. P. intermedia KCOM 1107 균주 유전체 염기서열을 해독하여 보고한다.

• Journal of Periodontal and Implant Science
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• 제39권2호
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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.

• Restorative Dentistry and Endodontics
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• 제22권1호
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• pp.413-427
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• 1997

Porphyromonas endodontalis and Prevotella intermedia are black-pigmented anaerobic gram negative rods which have been isolated from infected root canals and submucous abscesses of endodontic origin. And they are associated with clinical symptoms such as pain, percussion, and foul odor. It has been reported that there are 3 serotypes according to capsule membrane in P. endodontalis and 2 DNA homology groups and 3 serotypes in P. intermedia, but there is no data available regarding genetic diversity for the species P. endodontalis and P. intermedia. The purpose of this study is to investigate genetic diversities between individual strains of P. endodontalis and P. intermedia which are indistinguishable by serotyping and biotyping using bacterial DNA restriction endonuclease analysis. 45 teeth with at least one clinical symptoms, with single canal, and with pulp necrosis were sampled. For sampling bacteria, access cavity was prepared after disinfecting tooth and its surroundings. Then the paper point was inserted to the apex of the canal, leave there for 15 seconds, and finally it was placed into PRAS Ringer's solution and PBS solution. P. endodontalis and P. intermedia were identified by biochemical test and IIF after subculturing black and brown colonies which were produced after 7 days of incubation on BAP in anaerobic chamber. P. endodontalis and P. intermedia strains were grown in BHI broth and whole genomic DNA was extracted by phenol-chloroform extraction technique and digested by restriction endonuclease, Eco RI and Pst I. The resulting DNA fragments were separated by agarose gel electrophoresis, stained with EtBr and photographed under UV light. The results were as follows : 1. In both P. endodontalis and P. intermedia, different serotypes could be found within a root canal of same patient. 2. There were obvious genetic heterogeneity within a patient and within a serotype in both P. endodontalis and P. intermedia. 3. P. endodontalis serotype c, isolated from different patients, exhibited limited genotypic diversity.

• Journal of Periodontal and Implant Science
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• 제25권1호
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• pp.89-98
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• 1995

P. intermedia are considered an important pathogen in adult periodontitis, rapidly progressing periodontitis, refractory periodontitis, pregnancy gingivitis, acute necrotizing ulcerative gingivitis, pubertal gingivitis. So far 2 DNA homology groups and 3 serotypes of P. intermedia have been reported but there is no data available as yet regarding genetic diversity for the species P. intermedia. The purpose of this study is to investigate, using bacterial DNA restriction endonuclease analysis, genetic diversity between individual strains of P. intermedia which are indistinguishable by serotyping and biotyping, occurrence of an intrafamilial transmission and genetic heterogeneity between P. intermedia strains isolated within a patient and within the same serotypes. The families who have had no systemic disease, no experience of periodontal treatment for the previous 1 year and no experience of antibiotics for the previous 6 months were selected and subgingival plaque was collected at 4 sites in each person and incubated in the anaerobic chamber. P. intermedia were identified by colony shape, gram stain, biochemical test, SK-I03(Sunstar Inc.) test and IIF using monoclonal antibody was perfomed for the determination of serotypes. P. intermedia strains were grown in BHI broth and whole genomic DNA was extracted and digested by restriction endonuclease. The resulting DNA fragments were separated by agarose gel electrophoresis, stained and photographed under UV. As the results of this study, intrafamilial vertial transmissions could be assessed in 2 families and horizintal transmissions in another 2 families. There were different DNA digest patterns within a patient, so P. intermedia showed that individuals could be colonized by multiple clonal types at anyone time. And different serotypes could be found within a patient and in the same serotype within a patient, obvius genetic heterogeneity could not be assessed. But in the same serotype in different famies, there were differences in the DNA digest patterns.

• Journal of Periodontal and Implant Science
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• 제40권3호
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• pp.119-124
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• 2010

Purpose: In addition to regulating body weight, leptin is also recognized for its role in the regulation of immune function and inflammation. The purpose of this study was to investigate the effect of leptin on Prevotella (P.) intermedia lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-$\alpha$ production in differentiated THP-1 cells, a human monocytic cell line. Methods: LPS from P. intermedia ATCC 25611 was 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. The amount of TNF-$\alpha$ and interleukin-8 secreted into the culture medium was determined by enzyme-linked immunosorbent assay (ELISA). TNF-$\alpha$ and Ob-R mRNA expression levels were determined by semi-quantitative reverse transcription-polymerase chain reaction analysis. Results: Leptin enhanced P. intermedia LPS-induced TNF-$\alpha$ production in a dose-dependent manner. Leptin modulated P. intermedia LPS-induced TNF-$\alpha$ expression predominantly at the transcriptional level. Effect of leptin on P. intermedia LPS-induced TNF-$\alpha$ production was not mediated by the leptin receptor. Conclusions: The ability of leptin to enhance P. intermedia LPS-induced TNF-$\alpha$ production may be important in the establishment of chronic lesion accompanied by osseous tissue destruction observed in inflammatory periodontal disease.

• 한국미생물·생명공학회지
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• 제31권2호
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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.

• Animal Systematics, Evolution and Diversity
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• 제30권1호
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• pp.44-48
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• 2014

As a result of continuous taxonomic investigations of Korean crabs, Pugettia intermedia Sakai, 1938 is newly reported from Korean waters. Pugettia intermedia had previously been reported in Korean fauna, but the previous reports of P. intermedia were resulted in misidentification of P. quadridens (De Haan, 1839). Pugettia intermedia differs from P. quadridens in having two subequal medial lobes of the first pleopod in male. In P. quadridens, one of the two medial lobes is about half-length of the other. Pugettia intermedia occurs on the southern coast of the Korean peninsula. The descriptions and illustrations of this species are provided herein.

• 대한소아치과학회지
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• 제33권2호
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• pp.304-310
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• 2006

본 연구는 치주질환 주요 병인균주 중의 하나인 P. intermedia를 대상으로 하여, 이 균주에서의 hemin 결합 단백질 유전자를 분리하고 염기서열을 결정하기 위하여 수행되었다. 본 연구에서는 약 5,000개의 recombinant colony들을 스크리닝하여 hemin 결합 단백질 유전자를 포함하고 있는 것으로 여겨지는 1개의 클론(pHem1)을 확인하였다. Restriction enzyme mapping 결과 pHem1의 insert DNA의 크기는 약 2.5kb이었으며 P. intermedia chromosomal DNA 내에는 hemin 결합 단백질 유전자가 single copy로 존재하였고, transcript의 크기는 약 1.8kb이었다. 분리한 pHem1 유전자는 1개의 ORF를 가지고 있으며, ORF의 크기는 2,550bp로 약 850개의 아미노산 폴리펩타이드로 구성되어 있다. 또한, pHem1 유전자는 이미 밟혀진 다른 유전자들과 상동성을 보이지 않았다. 본 연구는 P. intermedia에서의 porphyrin생리 및 hemin 획득기전을 분자생물학적으로 규명하는데 있어 중요한 의의가 있으리라 사료된다. 향후 pHem1 유전자의 특성 분석 등이 수행되어야 할 것으로 여겨진다.