• Journal of Microbiology and Biotechnology
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• v.19 no.3
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• pp.314-322
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• 2009

Four different bacterial colonies were isolated from an old stock of an entomopathogenic nematode, Steinernema monticolum. They all showed entomopathogenicity to final instar larvae of beet armyworm, Spodoptera exigua, by hemocoelic injection. However, they varied in colony form, susceptibility to antibiotics, and postmortem change of the infected host insects. Biolog microbial identification and 16S rDNA sequence analyses indicate that these are four different species classified into different bacterial genera. Owing to high entomopathogenicity and a cadaver color of infected insect host, Serratia sp. was selected as a main symbiotic bacterial species and analyzed for its pathogenicity. Although no virulence of Serratia sp. was detected at oral administration, the bacteria gave significant synergistic pathogenicity to fifth instar S. exigua when it was treated along with a spore-forming entomopathogenic bacterium, Bacillus thuringiensis. The synergistic effect was explained by an immunosuppressive effect of Serratia sp. by its high cytotoxic effect on hemocytes of S. exigua, because Serratia sp. caused septicemia of S. exigua when the bacterial cells were injected into S. exigua hemocoel. The cytotoxic factor(s) was present in the culture medium because the sterilized culture broth possessed high potency in the cytotoxicity, which was specific to granular cells and plasmatocytes, two main immune-associated hemocytes in insects.

• Journal of Periodontal and Implant Science
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• v.33 no.4
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• pp.585-597
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• 2003

Porphyromonas gingivalis has been implicated as an important periodontophathic bacterium in the etiology and progression of periodontal diseases. It has been reported that P.gingivalis may mediate periodontal destruction not only directly through its virulence factors, but also indirectly by including complex host mediated inflammatory reponses. The purpose of this study was t o evaluate the effects of P.gingivalis on the bone formation and resorption by osteoblasts. For this purpose, after determining the concentration below which sonicated P.gingivalis extracts (SPEs) have no cytotoxicity on mouse calvarial primary osteoblastic (POB) cells, we investigated the effects of SPEs on the alkaline phosphatase (ALP) activity, matrix metalloproteinase (MMP) expression (MMP-2, -9, 13), and prostaglandin $E_2$ ($PGE_2$) release in POB cells by treatment with SPEs below that concentration. The results were as follows; 1. SPEs showed no cytotoxic effect on POB cells up to a concentration of 1 ${\mu}m$/ml. 2. The treatment with SPEs reduced ALP activity in a dose-dependent manner in POB cells, In addition, when we investigated the effect of SPEs (1 ${\mu}m$/ml) on ALP activity for different exposure periods, statistically significant inhibition of ALP activity was shown at 2 days of exposure, and further significant inhibition occurred by extending the periods of exposure. 3. The treatment with SPEs stimulated the gene expression of MMP-9 in POB cells. 4. The pre-treatment with SPEs increased the amount of $PGE_2$ released in POB cells. In summary, the present study shows that P.gingivalis could inhibit osteogenesis and stimulate bone resorption not only by reducing ALP activity but also by increasing MMP-9 mRNA expression in osteoblasts, possibly through an endogenous $PGE_2$ pathway. In addition, our results suggest that if P.gingivalis affects osteoblasts in early differentiation stage, such effects by P. gingivalis could be irreversible.

• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.4
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• pp.587-596
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• 2006

Most of oral streptococci express the Antigen I/II (AgI/II) proteins, cell wall anchored adhesions. AgI/II protein binds to salivary agglutinin glycoprotein, a component of tooth pellicle and to ligands in other bacteria. These associations play important roles in bacterial colonization. Recently, it was reported that diverse host molecules also interact with AgI/II protein and that these interactions induce inflammatory responses from host cells. Among mutans streptococci containing -type hemolytic activity, Streptococcus mutans is a causative agent for dental caries. Compared with many other strains of S. mutans, GS-5 strain is unique in that this bacterium expresses truncated secretory AgI/II protein due to the nonsense mutation in the agI/II gene. This indicates that S. mutans GS-5 has a different clinical role and a recent report supported this idea based on the results from clinically isolated S. mutans strains. Previously, we had cloned agI/II gene from S. mutans GS-5 and generated recombinant N-terminal AgI/II protein. In this study, we further produced a hybridoma line expressing anti-AgI/II monoclonal antibodies named as 1C11A. This antibody showed high sensitivity to AgI/II protein in Western blot and ELISA. This new reagent will provide a basis for investigating the mechanisms of AgI/II-related diseases.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.4
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• pp.660-666
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• 2003

The purpose of this study was to investigate the inhibitory effect of Er:YAG laser against the intraoral acid producing bacterium of S. mutans. Bacterial pellet containing S. mutans KCTC 3065 was irradiated by Er:YAG laser having a $650\;{\mu}m$ diameter beam by non-contact mode. Irradiated parameters were 50mJ, 10Hz and exposure time were 1s, 3s, 5s, 7s, 9s respectively. We obtained the following results of relative growth rate and acid-producing ability of S. mutans by culturing for 48hrs. 1. The growth rate of S. mutans was decreased in the group of laser irradiation compared to the control group(P<0.01). 2. The growth rate at laser irradiation group of 7s, 9s irradiation time was decreased significantly compared to the laser irradiation group of 1s, 3s, 5s irradiation time, until 12 hours(P<0.05). After 24 hours, all groups of laser irradiation were not found to be statistically different in each other. 3. The acid-producing ability of S. mutans was inhibited for a certain duration by irradiation of laser. In summary, the growth rate and acid producing ability of S. mutans decreased according to laser irradiation. This effect was directly related to the amount of irradiation time. These results suggested that Er:YAG laser had an growth inhibition effect on S. mutans.

• Restorative Dentistry and Endodontics
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• v.23 no.1
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• pp.43-53
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• 1998

Dental caries is induced by organic acids produced by oral bacteria. In order to prevent dental caries, therefore, it is essential to maintain neutral pH in the oral cavity. Urea plays a major role in oral pH homeostasis. Urea is hydrolyzed by bacterial ureases to ammonia, causing a pH elevation. Streptococcus salivarius has been shown to be a major contribution to oral ureolysis. Synthesis of urease by S. salivarius appears to be constituitive, but can be greatly enhanced by low pH. It is, therefore, conceivable that ureolytic activity of S. salivarius from a carious lesion is greater than that of the bacterium from a healthy tooth. In the present study, urease activity of S. salivarius isolates from dental plaque of carious lesions was compared with that of the isolates from plaques of the teeth and the dorsum of the tongue; 45 S. salivarius strains were isofated from carious lesions(>C2) of 21 individuals with dental caries and 30 strains from 10 individuals without dental caries. The results were as follows: 1. All the 21 individuals with dental caries harbored ureolytic S. salivarius whereas 3 of 13 individuals without dental caries harbored non-ureolytic strains of S. salivarius. 2. All the 45 S. saliuarius isolates from carious lesions showed urease activity. In contrast, of 30 isolates from individuals without dental caries, 17 isolates(56.7%) did not demonstrate urease activity, or if any, very little(<5${\mu}mol$/min/mg). 3. Urease activity of the isolates from carious lesions was greater than that of the isolates from individuals without dental caries : the urease activity ranged from 42 to $381{\mu}mol$/min/mg and from 0 to $208{\mu}mol$/min/mg, respectively. 4. At acid pH(5.5), the isolates which showed intermediate urease activity at pH 7.0 demonstrated even higher activity whereas the isolate with no or lower urease activity did not show any significant difference in their activity. However, the isolates with the greatest urease activity from both individuals with and without dental caries, exhibited a rather much lower urease activity at pH 5.5. The overall results suggest that isolates may have their own urease activity but the isolates exposed to chronic acidic environment of the carious lesion might elevate urease activity of S. salivarius, which in turn, might influence on survival of S. salivarius itself and other bacteria, establishing a new oral bacterial ecosystem.

• KSBB Journal
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• v.26 no.6
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• pp.513-516
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• 2011

The occurrence of dental caries is mainly associated with oral pathogens, especially cariogenic Streptococcus mutans. The aim of this study was to isolate and characterize lactic acid bacterium showing inhibitory activity against cariogenic Streptococcus mutans. As results, an isolate with strong inhibitory activity was obtained from Kimchi and it was identified as Lactobacillus sakei by API and 16S rRNA gene analyses. This strain secreted an inhibitory compound in cell growth medium and the activity of the compound was completely disappeared by proteinase K revealing the fact that the compound is proteinous substance, bacteriocin. Optimal culture condition for bacteriocin production by Lb. sakei K-7 was at pH 7.5 and $37^{\circ}C$ for 18 h. Oral administration of this isolate may give anticariogenic and probiotic effects on hosts.

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1829-1835
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• 2016

Dental caries is caused by cariogenic biofilm, an oral biofilm including Streptococcus mutans. Recently, the prevention of dental caries using various probiotics has been attempted. Lactococcus lactis HY 449 is a probiotic bacterium. The aim of this study was to investigate the effect of L. lactis HY 449 on cariogenic biofilm and to analyze its inhibitory mechanisms. Cariogenic biofilm was formed in the presence or absence of L. lactis HY 449 and L. lactis ATCC 19435, and analyzed with a confocal laser microscope. The formation of cariogenic biofilm was reduced in cultures spiked with both L. lactis strains, and L. lactis HY 449 exhibited more inhibitory effects than L. lactis ATCC 19435. In order to analyze and to compare the inhibitory mechanisms, the antibacterial activity of the spent culture medium from both L. lactis strains against S. mutans was investigated, and the expression of glucosyltransferases (gtfs) of S. mutans was then analyzed by real-time RT-PCR. In addition, the sucrose fermentation ability of both L. lactis strains was examined. Both L. lactis strains showed antibacterial activity and inhibited the expression of gtfs, a nd t he d ifference b etween both strains did not show. In the case of sucrose-fermenting ability, L. lactis HY 449 fermented sucrose but L. lactis ATCC 19435 did not. L. lactis HY 449 inhibited the uptake of sucrose and the gtfs expression of S. mutans, whereby the development of cariogenic biofilm may be inhibited. In conclusion, L. lactis HY 449 may be a useful probiotic for the prevention of dental caries.

• Korean Journal of Microbiology
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• v.55 no.1
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• pp.58-60
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• 2019

Anaerobic Gram-negative bacterium Prevotella intermedia is a part of normal flora of the oral cavity and associated with various types of oral and systemic diseases. We present here a draft genome sequence of P. intermedia ATCC 15032, originally isolated from cervicofacial actinomycosis. The genome is 2,848,426 bp in length and has a GC content of 43.45%. The genome includes 2,358 protein-coding genes, 5 rRNAs, and 43 tRNA. The sequence information will provide important clues in understanding the genome diversity within the bacterial species, and genetic basis for phenotypic differences among P. intermedia strains.

• International Journal of Oral Biology
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• v.33 no.4
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• pp.163-177
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• 2008

Periodontal disease, a form of chronic inflammatory bacterial infectious disease, is known to be a risk factor for cardiovascular disease (CVD). Porphyromonas gingivalis has been implicated in periodontal disease and widely studied for its role in the pathogenesis of CVD. A previous study demonstrating that periodontopathic P. gingivalis is involved in CVD showed that invasion of endothelial cells by the bacterium is accompanied by an increase in cytokine production, which may result in vascular atherosclerotic changes. The present study was performed in order to further elucidate the role of P. gingivalis in the process of atherosclerosis and CVD. For this purpose, invasion of human aortic smooth muscle cells (HASMC) by P. gingivalis 381 and its isogenic mutants of KDP150 ($fimA^-$), CW120 ($ppk^-$) and KS7 ($relA^-$) was assessed using a metronidazole protection assay. Wild type P. gingivalis invaded HASMCs with an efficiency of 0.12%. In contrast, KDP150 failed to demonstrate any invasive ability. CW120 and KS7 showed relatively higher invasion efficiencies, but results for these variants were still negligible when compared to the wild type invasiveness. These results suggest that fimbriae are required for invasion and that energy metabolism in association with regulatory genes involved in stress and stringent response may also be important for this process. ELISA assays revealed that the invasive P. gingivalis 381 increased production of the proinflammatory cytokine interleukin (IL)-$1{\beta}$ and the chemotactic cytokines (chemokine) IL (interleukin)-8 and monocyte chemotactic (MCP) protein-1 during the 30-90 min incubation periods (P<0.05). Expression of RANTES (regulation upon activation, normal T cell expressed and secreted) and Toll-like receptor (TLR)-4, a pattern recognition receptor (PRR), was increased in HASMCs infected with P. gingivalis 381 by RT-PCR analysis. P. gingivalis infection did not alter interferon-$\gamma$-inducible protein-10 expression in HASMCs. HASMC nonspecific necrosis and apoptotic cell death were measured by lactate dehydrogenase (LDH) and caspase activity assays, respectively. LDH release from HASMCs and HAMC caspase activity were significantly higher after a 90 min incubation with P. gingivalis 381. Taken together, P. gingivalis invasion of HASMCs induces inflammatory cytokine production, apoptotic cell death, and expression of TLR-4, a PRR which may react with the bacterial molecules and induce the expression of the chemokines IL-8, MCP-1 and RANTES. Overall, these results suggest that invasive P. gingivalis may participate in the pathogenesis of atherosclerosis, leading to CVD.

• Journal of Periodontal and Implant Science
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• v.29 no.3
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• pp.663-676
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• 1999

Porphyromonas gingivalis, a Gram negative, anaerobic, asaccharolytic rod, is one of the most frequently implicated pathogens in human periodontal disease and has a requirement for hemin for growth. A 30 kDa (heated 24 kDa) hemin-binding protein whose expression is both hemin and iron regulated has recently been purified and characterized in this oral pathogen. This study has identified a hemin-binding P. gingivalis protein by expression of a P. gingivalis genomic library in Escherichia coli, a bacterium which does not require or transport exogenous hemin. A library of genomic DNA fragments from P. gingivalis was constructed in plasmid pUC18, transformed into Escherichia coli strain $DH5{\alpha}$ , and screened for recombinant clones with hemin-binding activity by plating onto hemin-containing agar. Of approximately 10,000 recombinant E. coli colonies screened on LB-amp-hemin agar, 10 exhibited a clearly pigmented phenotype. Each clone contained various insert DNA. The Hind III fragment transferred to the T7 RNA polymerase/promoter expression vector system produced a sligltly smaller (21 kDa) protein, a precursor form, immunoreactive to the antibody against the 24 kDa protein, suggesting that the cloned DNA fragment probably carried an entire gene for the 24 kDa hemin-binding protein.