• Title/Summary/Keyword: Actinobacillus

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Biological Characterization of the Omp1-like Protein from Actinobacillus actinomycetemcomitans

  • Ha, Jung-Hye;Jeong, Mi-Suk;Jo, Wol-Soon;Jeong, Min-Ho;Jang, Se-Bok
    • Bulletin of the Korean Chemical Society
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    • v.31 no.2
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    • pp.275-280
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    • 2010
  • Actinobacillus actinomycetemcomitans is a gram-negative, nonmotile coccobacillus bacterium that is associated with several human diseases, including endocarditis, meningitis, osteomyelitis, subcutaneous abscesses and periodontal diseases. A full-length Omp1-like protein gene from A. actinomycetemcomitans was cloned into a pQE30 vector and overexpressed in Escherichia coli BL21(DE3) cells. The protein revealed sequence homologies to Seventeen kilodalton proteins (Skp) from Pasteurella multocida and E. coli that have been characterized as periplasmic chaperones. This soluble Omp1-like protein was successfully purified to homogeneity for further folding and functional studies. The purity, identity, and conformation of the protein were determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis, matrix-assisted laser desorption ionization mass spectrometry, circular dichroism, fluorescence spectroscopic, and differential scanning calorimetric studies. We showed that the protein formed an oligomer larger than a tetramer. We found, further, that it is comprised of mostly $\alpha$-helices and boasts high thermal stability.

Multiplex Real-Time PCR for Simultaneous Detection of 6 Periodontopathic Bacteria (Multiplex Real-Time PCR을 이용하여 6종의 주요 잇몸질환 유발 미생물을 동시에 검출하는 기법)

  • Cho, Hong-Bum
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.292-296
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    • 2013
  • This study utilized an analysis method for detecting six microorganisms, such as Actinobacillus actinomycetemcomitans, Campylobacter rectus, Porphyromonas gingivalis, Tannerella forsythus, Treponema denticola, and Prevotella intermedia, triggering periodontal disease, using multiplex real-time polymerase chain reaction (PCR). The analysis including internal control was made by dividing the six species into two groups using four fluorescence dyes, and it was verified that there was no interference or cross-reaction between the target species and different kinds of oral microbial species. Qualitative and quantitative analyses were conducted on each microorganism in various samples, such as saliva and the plaque, using the multiplex real-time PCR and comparative analysis between periodontitis patients and healthy people, revealing obvious differences between them.

Antibiotic Susceptibility Of Actinobacillus Actinomycetemcomitans From The Patients With Adult Periodontitis (성인형 치주염 환자에서 분린한 Actinobacillus actinomycetemcomitans의 항생재 내성에 관한 연구)

  • Na, Yong-Cheol;You, Hyung-Keun;Sihn, Hyung-Sik
    • Journal of Periodontal and Implant Science
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    • v.26 no.1
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    • pp.202-215
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    • 1996
  • The purpose of this study was to an in estigate the antibiotic susceptibiliity of Actinoobacillus actinomycetemcomitans isolated from the subgingival plaque to adult periodontitis. Seven bacterial strains were tested for their susceptibility to 10 antimicrobial agents under disk diffusion method and broth dilution methold, Seven patients with deep pocket(6mm) were selected for this study. They had not taken antibiotics for 6 months and no history of dental treatment for 6 months and no history of dental treatment for 6 months before this study. The results were as follows : 1. For the antibiotic disk diffusion method, seven A. actinimycetemcomitans were tested with 10 antimicrobial agents which comprised penicillin, gentamycin, clindamycin, lincomycin, ampicillin, erythromycin, tetracycline, amikacin, chloramphenicol, and vancomycin. The sensitive antibiotics were tetracycline, vancomycin, and chloramphenicol and the resistant antibiotics were clindamycin, and lincomycin. The other antimicrobial agents were less active. 2. From the study of determination on the minimal inhibitory concentration(MIC) by broth dilution method, the MIC of tetracycline to seven strains of the A. actinomycetemcomitans was $0.5-1.0{\mu}g/ml$, that of clindamycin was $32{\mu}g/ml$. These data suggest that tetracycline may be valuable drugs in the elimination of A. actinimycetemcomitans from the patients with adult periodontitis

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Identification of Actinobacillus actinomycetemcomitans Using Species-Specific 16S rDNA Primers

  • Kim Su Gwan;Kim Soo Heung;Kim Mi Kwang;Kim Hwa Sook;Kook Joong Ki
    • Journal of Microbiology
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    • v.43 no.2
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    • pp.209-212
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    • 2005
  • The purpose of this study was to develop species-specific PCR primers for use in the identification and detection of Actinobacillus actinomycetemcomitans. These primers target variable regions of the 168 ribosomal RNA coding gene (rDNA). We assessed the specificity of the primers against 9 A. actinomycetemcomitans strains and 11 strains (3 species) of the Haemophilus genus. Primer sensitivity was determined by testing serial dilutions of the purified genomic DNAs of A. actinomycetemcomitans ATCC$ 33384^$T Our obtained data revealed that we had obtained species-specific amplicons for all of the tested A. actinomycetemcomitans strains, and that none of these amplicons occurred in any of the other species. Our PCR protocol proved able to detect as little as 4 fg of A. actinomycetemcomitans chromosomal DNA. Our findings suggest that these PCR primers are incredibly sensitive, and should prove suitable for application in epidemiological studies, as well as the diagnosis and monitoring of periodontal pathogens after treatment for periodontitis.

A STUDY ON THE RELATIONSHIP BETWEEN THE CLINICAL SYMPTOMS AND THE PREDOMINENT PATHOGENIC MICROFLORA IN INFECTED ROOT CANALS (감염 근관에서의 주요 병인균과 임상증상간의 관계에 관한 연구)

  • Kim, Min-Kyum;Yoon, Soo-Han
    • Restorative Dentistry and Endodontics
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    • v.14 no.1
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    • pp.85-96
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    • 1989
  • In the Infected root canals, the majority of the isolated bacteria are either strict anaerobes or microaerophilic organisms. Among thease, Black-pigmented Bacteroides species are the most important and have relation with clinical symptoms. The purpose of this study were to investigate on the 7 different types of bacteria which include Black-pigmented Bacteroides - Black--pigmented Bacteroides, Actinobacillus actinomycetemcomitans, Streptococcus, Capnocytophaga, Eiknella corrodens, Fusobacterium, Actinomyces - and the interrelationship between these 7 bacterical species, and to compare Black-pigmented Bacteroides with the clinical symptoms in infected root canals. The canal contents of 15 necrotized teeth with 8 clinical symptoms were sampled and cultured aerobically and anaerobically for growth in 7 selective agar plates for 7 bacterial species. Black-pigmented Bacteroides and Streptococcus subspecies were identified by biochemical tests. The results were as follows; 1. 70.51% of the bacteria isolated were anaerobes and 29.49% were aerobes. 2. B. loescheii, B. intermedius, B. denticola were isolated in 8 cases, Streptococcus was isolated in 8 cases, Fusobacterium was isolated in 6 cases, Actinomyces was isolated in 5 cases and Eiknella corrodens, Actinobacillus actinomycetemcomitans, Capnocytophaga were not isolated. 3. There were mutual inhibition of growth between Black-pigmented Bacteroides and Streptococcus. (P<0.01) But Actinomyces and Fusobacterium showed mutual aids for growth. (P<0.05) 4. Black-Pigmented Bacteroides was found to be related both to sinus tract formation and to apical sensitivity to palpation.(P < 0.05).

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A study on the practicability of genotyping and cytotoxicity of Actinobacillus actinomycetemcomitans isolated from periodontal patients (Actinobacillys actinomycetemcomitans의 유전자형의 유용성과 세포독성에 관한 연구)

  • 조월순;정민호;이상화;황희성
    • Journal of Life Science
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    • v.11 no.1
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    • pp.8-18
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    • 2001
  • The purpose of this study was to evaluate the genotypic characterization of Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) using arbitrarily primed polymerase chain reaction (AP-PCR), to investigate the cytotoxicity of both clinical isolates and standard strains of A. actinomycetemcomitans for the human Jurkat T cells, and to measure the osteoclastogenic cytokines released by Jurkat cells infected with these bacterial strains. The random sequence primer 15 and 16 could distinguish different AP-PCR profiles between clinical isolates of A. actinomycetemcomitans. A. actinomycetemcomitans significantly suppressed Jurkat cell viability in time dependent fashion and the results of DNA fragmentation assay indicated that this bacterial species induced apoptosis in Jurkat cells undergoing apoptosis released the osteoclastogenic cytokine, IL-1$\beta$, IL-6, TNF-$\alpha$. These data support the hypothesis that induction of apoptosis is at least one essential step in A. actinomycetemcomitans induced local immunosuppressive pathway, and that A. actinomycetemcomitans can modulate the immunomodulatory cell population in the periodontal tissue by inducing T cell death through apoptosis, and that apoptosis of local resident T cells may play an active role in bone resorption by releasing osteoclastogenic cytokines, e.g. IL-1$\beta$, IL-6, TNF-$\alpha$.

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The Distribution of Actinobacillus actinomycetemcomitans, Hemophilus aphrophilus and Hemophilus paraphrophilus in Subgingival Plaque and Saliva from Korean Periodontitis Patients using PCR (PCR을 이용한 치주환자의 타액과 치은연하치태의 세균분포에 관한 연구)

  • Choi, Ok-Sun;Ahn, Gwang-Sook;Kim, Hye-Jin;Lee, Eun-Sook
    • Journal of Korean society of Dental Hygiene
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    • v.2 no.2
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    • pp.201-213
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    • 2002
  • The closely related species Actinobacillus actinomycetemcomitans, Hemophilus aphrophilus and Hemophilus paraphrophilus are common findings in oral microbiota. The aims of this study were to compare the distribution of three species in healthy subjects and periodontitis patients using PCR for 16s rRNA gene. The DNA was extracted from the subgingival plaque and saliva in 122 subjects for restriction enzyme analysis with Hinf I and Hha I. In case of periodontally healthy person, A. actinomycetemcomitans was predominant than H. paraphrophilus in saliva sample, but H. paraphrophilus was predominant than A. actinomycetemcomitans in subgingival plaque sample. On the contrary, in case of periodontitis patients, H. paraphrophilus was predominant than A. actinomycetemcomitans in saliva sample, but A. actinomycetemcomitans was predominant than H. paraphrophilus in subgingival plaque sample. In addition, the fact was confirmed that the distribution of A. actinomycetemcomitns of women periodontitis patients was somewhat higher than men periodontitis patients in saliva and subgingival plaque samples. We convinced that the PCR method for 16s rRNA gene was important for screening and monitoring of periodontal disease.

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