• Journal of Microbiology and Biotechnology
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• v.15 no.3
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• pp.519-524
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• 2005

An ${\alpha}$-L-rhamnosidase (EC 3.2.1.40.), which transforms quercitrin to quercetin, was purified from Fusobacterium K-60, a human intestinal anaerobic bacterium. The specific activity of the purified ${\alpha}$-L-rhamnosidase was 2.89 mol/min/mg protein. ${\alpha}$-L-Rhamnosidase, whose molecular size was 170 kDa by gel filtration, was composed of four subunits ($M_r$ 41,000 Da) with pI and optimal pH values of 5.2 and 5.5-7.0, respectively. The apparent $K_m$ and $V_{max}$ values for p-nitrophenyl-${\alpha}$-L-rhamnopyranoside and quercitrin were determined to be 0.057 mM and 3.4 mol/min/mg, and 0.077 mM and 5.0 mol/min/mg, respectively. This enzyme was strongly inhibited by $Cu^{2+},\;Mn^{2+}$, L-rhamnose, and p-chlormercuriphenylsulfonic acid. These findings suggest that the biochemical properties and substrate specificity of the purified enzyme are different from those of the previously purified ${\alpha}$-L-rhamnosidase. This is the first reported purification of quercitrin-hydrolyzing ${\alpha}$-L-rhamnosidase from intestinal bacteria.

• Proceedings of the PSK Conference
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• 2000.04a
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• pp.154.2-154.2
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• 2000

• Journal of the korean academy of Pediatric Dentistry
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• v.32 no.2
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• pp.344-356
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• 2005

There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic acid bacteria were isolated from inhabitants of caries-free children's oral cavity, which inhibited the formation of artificial plaque by Streptococcus mutans and the production of volatile sulfur compounds by anaerobic bacteria. The isolates were identified by the test using API 50 CHL medium kit and 16S rDNA partial sequencing. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. When Streptococcus mutans was cultured in the media, the mean weight of formed artificial plaque on the orthodontic wires was $124.4{\pm}30.4\;mg$, whereas being reduced to $5.2{\pm}2.0mg$ and $10.6{\pm}6.6mg$ in the media cultured with Streptococcus mutans and each isolate, respectively (p<0.05) 3. The number of viable cells of Streptococcus mutans was $3.4{\times}10^9$ per ml in the cultured solution, whereas those of Streptococcus mutans in the combined culture with each of isolates were $4.6{\times}10^8\;and\;2.4{\times}10^8$ per ml. 4. The optical density was 1.286 in the supernatant of Fusobacterium nucleatum after vortexing for 30minutes, whereas in the supernatant of combined Fusobacterium nucleatum and each isolate, they were reduced to 0.628 and 0.497, which the percentages of coaggregation between them were 29.4% and 57.8%, respectively 5. The optical density of Fusobacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$, being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusobacterium nucleatum and each isolate. The optical density of Porphyromonas gingivalis precipitate was 1.932 in the culture media, being reduced to 1.170 and 1.266 in the coaggregated precipitates of Porphyromonas gingivalis and each isolate. 6. When two isolates were tested with API 50 CHL medium kit, those were identified Lactobaciallius salivarius and Lactobaciallius delbrueckii subsp. lactis. 7. The similarity values of 16S rDNA sequence between each of isolates and Lactobaciallius salivarius subsp. salicinius were 99.60% and 99.73%, respectively, meaning that isolates were Lactobaciallius salivarius subsp. salicinius. These results indicated that two strains isolated from caries-free children's saliva, which inhibited the formation of artificial plaque and the production of volatile sulfur compounds, were identified as Lactobaciallius salivarius subsp. salicinius.

• Proceedings of the PSK Conference
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• 2001.04a
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• pp.180.1-180.1
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• 2001

• Journal of dental hygiene science
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• v.9 no.2
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• pp.249-255
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• 2009

This study was carried out for the purpose of comparing bacterial culture method, single PCR, and multiplex PCR for identification of F. nucleatum and A. actinomycetemcomitans in subgingival plaque of adult periodontitis. Targeting 20 patients with adult periodontitis, the subgingival plaque was collected in teeth, respectively, for #16, #36, #44. A bacillus was cultivated by painting it over the solid selective media of F. nucleatum and A. actinomycetemcomitans. Bacterial species were detected in 0 tooth with 12 pieces, respectively. Through single PCR and multiplex PCR, the positive reaction was indicated in 43 teeth with 45 pieces, respectively, as for F. nucleatum, and in 1 tooth with 4 pieces, respectively, as for A. actinomycetemcomitans. In the comparative analysis between bacterial identification methods. F. nucleatum showed the more statistically significant difference(p=0.0(0) in comparison between single PCR and multiplex PCR. Even A. actinomycetemcomitans was indicated significantly(p=0.067) in a case that is based on 0.1 in significant level in the comparison between single PCR and multiplex PCR. In conclusion, as a result of comparing the bacterial identification methods, the detection frequency was indicated to be higher in PCR than in bacterial culture method. Single PCR and multiplex PCR showed the mutually similar detection frequency. Accordingly, given thinking of economic efficiency, quickness, and reduction in labor force, it is thought to be more efficient method to use single PCR as the bacterial identification method.

• Restorative Dentistry and Endodontics
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• v.24 no.3
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• pp.490-494
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• 1999

The aim of this in vitro study was to investigate the coronal leakage of obligate anaerobes into root canals obturated with two different techniques. 48 extracted human teeth with straight, single root canals were prepared with crown-down technique with Profile under copious irrigation until the master apical file was size 40. The teeth were divided randomly into experimental groups (40 teeth) and control groups (8 teeth). In the experimental groups, 20 teeth were obturated with lateral condensation and other 20 teeth were obturated with continuous wave technique with System B. Coronal leakage of two root canal filing technique was evaluated using anaerobic bacterial leakage model with Fusobacterium nucleatum(ATCC 25586) for 60 days. The results were as follows 1. The incidence of bacterial leakage of experimental groups was 65% in group 1 (lateral condensation) and 60% in group 2 (continuous wave technique with System B). This difference was not statistically significant (P>0.05). 2. There was no statistically significant difference(P>0.05) in leakage score between group 1 (lateral condensation) and group2 (continuous wave technique with System B).

• Journal of Periodontal and Implant Science
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• v.40 no.2
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• pp.61-68
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• 2010

Purpose: The present study was performed to clarify the relationship between periodontal disease severity and selected immunological parameters consisting of serum IgG titer against periodontopathogenic bacteria, the expression of the helper T-cell cytokine by gingival mononuclear cells, and patients' immunoreactivity to cross-reactive heat shock protein (HSP) epitope peptide from P. gingivalis HSP60. Methods: Twenty-five patients with moderate periodontitis had their gingival connective tissue harvested of gingival mononuclear cells during an open flap debridement procedure and peripheral blood was drawn by venipuncture to collect serum. The mean level of interproximal alveolar bone was calculated to be used as an index for periodontal disease severity for a given patient. Each of selected immunologic parameters was subject to statistical management to seek their correlations with the severity of periodontal disease. Results: A significant correlation could not be identified between serum IgG titers against specific bacteria (Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, and Streptococcus mutans) and the severity of periodontal disease. Expression of interleukin (IL)-10 by gingival mononuclear cells was statistically significant in the group of patients who had higher levels of alveolar bone height. However, a similar correlation could not be demonstrated in cases for IL-4 or interferon-$\gamma$. Patients' serum reactivity to cross-reactive epitope peptide showed a significant correlation with the amount of alveolar bone. Conclusions: It was concluded that expression of IL-10 by gingival mononuclear cells and patients' sero-reactivity to the cross-reactive HSP peptide of P. gingivalis HSP60 were significantly correlated with alveolar bone height.

• Archives of Pharmacal Research
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• v.27 no.1
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• pp.61-67
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• 2004

When ginseng water extract was incubated at $60^{\circ}C$ in acidic conditions, its protopanaxadiol ginsenosides were transformed to ginsenoside $Rg_3$ and ${\Delta}^{20}$-ginsenoside $Rg_3$. However, protopanaxadiol glycoside ginsenosides $Rb_1, Rb_2$ and Rc isolated from ginseng were mostly not transformed to ginsenoside $Rg_3$ by the incubation in neutral condition. The transformation of these ginsenosides to ginsenoside $Rg_3$ and ${\Delta}^{20}$-ginsenoside $Rg_3$ was increased by increasing incubation temperature and time in acidic condition: the optimal incubation time and temperature for this transformation was 5 h and $60^{\circ}C$ resepectively. The transformed ginsenoside $Rg_3$ and ${\Delta}^{20}$-ginsenoside $Rg_3$ were metabolized to ginsenoside $Rh_2$ and $\Delta^{20}$--ginsenoside $Rh_2$, respectively, by human fecal microflora. Among the bacteria isolated from human fecal microflora, Bacteroides sp., and Bifidobacterium sp. and Fusobacterium sp. potently transformed ginsenoside $Rg_3$ to ginsenoside $Rh_2$. Acid-treated ginseng (AG) extract, fermented AG extract, ginsenoside $Rh_2$ and protopanaxadiol showed potent cytotoxicity against tumor cell lines. AG extract, fermented AG extract and protopanaxadiol potently inhibited the growth of Helicobacter pylori.

• Restorative Dentistry and Endodontics
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• v.25 no.2
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• pp.235-242
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• 2000

The purpose of this in vitro study was to evaluate the sealing ability of three sealers(Sealapex, Pulp canal sealer, AH26) used with continuous wave method using an anaerobic bacterial leakage model. 53 extracted human teeth with straight and single canals were prepared with crown-down pressureless technique using .04, .06 taper Profile(Maillefer, Swiss). Master apical file was maintained as #35 K-file. All canals of the experimental teeth were obturated with continuous wave method using System B(Analytic technology, U.S.A.) The teeth were randomly divided into three experimental groups of 15 and two control groups of 4. Experimental group 1 was obturated with Sealapex and group 2 with Pulp canal sealer, and group 3 with AH26. A dual chamber anaerobic bacterial leakage model was assembled. Brain heart infusion with yeast extract, hemin, menadion, and the chromogenic indicator bromocresol purple was used as the culture broth for Fusobacterium nucleatum(VPI 10197), The specimens were incubated in anaerobic chamber at $37^{\circ}C$ and were observed every 2 to 3 clays, The coronal leakage was evaluated through the color change of culture broth in lower chamber for 60 days. The results were as follows: 1. The incidence of bacterial leakage in group 1 (Sealapex group was 80%, 53% in group 2 (Pulp canal sealer), 27% in group 3 (AH26). 2. There were statistically significant differences in leakage scores between group 1 and group 2, and between group 1 and group 3, respectively. (P<0.05) 3. There was no significantly difference in leakage score between group 2 and group 3. (P>0 05)

• Restorative Dentistry and Endodontics
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• v.29 no.1
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• pp.51-57
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• 2004

본 연구는 에폭시레진계 봉함제 (AH26)과 두 가지의 상아질 접착제와 함께 근관충전을 시 행하였을 때와 에폭시레진계 봉함제 단독으로 사용하였을 때의 미세누출을 혐기성 세균모델을 이용하여 평가하였다. 52개의 단근치를 이용하여 .04, .06 taper Profile (Dentsply-Maillefer, Ballaigues, Swiss)을 사용하여 변형된 crown-down pressureless법으로 근관형성 하였다. 형성된 치근을 12개씩 무작위로 나누어 4개의 실험군으로 하였으며, 나머지 치아 중 2개를 양성대조군에, 2개는 음성대조군으로 사용하였다. 제1군은 All Bond 2(Bisco, Itasca, IL, USA)를 적용하고 거타퍼쳐와 AH26 (Dentsply, Konstanz, Germany)을 이용하여 continuous wave of condensation technique으로 근관충전 하였으며, 제 2군은Prime ＆ Bond NT (Dentsply, Konstanz, Germany)를 적용 후 거타퍼쳐와 AH26을 이용하여 충전하였으며, 제3군은 17％ EDTA를 적용하여 도말층을 제거한 후 거터퍼쳐 와 AH26을 사용하여 충전하였다. 제4군은 17％ EDTA를 적용하지 않고 거터퍼쳐와 AH26을 사용하여 충전하였다. Fusobacterium nucleatum (VPI 10197)을 추적자로 이용한 혐기성세균모델을 사용하여 혐기성배양기에서 배양시키면서 60일 동안 각군에 대한 미세누출 여부를 관찰하였다. 매일 배양액의 혼탁도와 색상변화를 관찰하여 기록하였다. 제4군에서 통계학적으로 유의할만한 미세누출을 가장 많이 보였으며(p<0.0005) 나머지 3개의 실험군에서는 서로간의 통계학적으로 유의할 만한 차이를 보이지 않았다. 주사전자현미경 관찰 시 제 1군과 제2군의 상아질 접착제가 상아세관으로 침투한 소견을 관찰 할 수 있었다.