• Journal of Microbiology and Biotechnology
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• 제15권3호
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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.

• 대한약학회:학술대회논문집
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• 대한약학회 2000년도 춘계총회 및 학술대회
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• pp.154.2-154.2
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• 2000

• 대한소아치과학회지
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• 제32권2호
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• pp.344-356
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• 2005

우리 인체에는 의학적으로 유용한 기능을 가진 세균들이 정상적으로 존재하며 구강내에도 독특한 기능을 나타내는 세균들이 상재한다. 본 연구에서는 치아우식증이 없는 소아의 타액에서 분리한 유산균 2주가 Streptococcus mutans에 의한 인공치태 형성과 혐기성 세균에 의한 휘발성 유황화합물 생성을 억제하는 것을 확인하고, API 50 CHL medium kit를 이용한 생화학적 검사와 16S rDNA sequencing으로 동정하여 다음과 같은 결과를 얻었다. 1. 분리균주는 2주 모두 그람양성 간균으로 과산화수소를 생성하였다. 2. 인공치태의 무게는 Streptococcus mutans의 단독 배양시 $124.4{\pm}30.4mg$이었으나, 분리균주와 병합 배양시에는 각각 $5.2{\pm}2.0mg,\;10.6{\pm}6.6mg$으로 현저하게 감소하였다(p<0.05). 3. Streptococcus mutans의 생균수는 단독 배양시 ml당 $3.4{\times}10^9$이었으나, 분리균주와 병합 배양시에는 각각 ml당 $4.6{\times}10^8$과 $2.4{\times}10^8$으로 감소하였다. 4. Fusobacterium nucleatum을 30분간 진탕한 후 측정한 상청액의 흡광도는 1.286이었으나, Fusobacterium nucleatum과 분리균주를 병합으로 30분간 진탕한 후 측정한 상청액의 흡광도는 각각 0.628과 0.497로 감소하였으며, 상호 결합 정도는 29.4%와 57.8%이었다. 5. Fusobacterium nucleatum의 단독 배양시 cysteine과 $FeSO_4$를 첨가한 배지를 가한 후 측정한 침전물의 배지 흡광도는 1.794이었으나, 분리균주와 병합 배양시 측정한 침전물의 배지 흡광도는 각각 1.144와 0.915로 감소하였으며, Porphyromonas gingivalis 단독 배양시 침전물의 배지 흡광도는 1.932이었으나 분리균주와 병합 배양시에는 침전물의 배지 흡광도가 각각 1.170과 1.266으로 감소하였다. 6. 분리균주를 API 50 CHL medium kit로 탄수화물 발효검사를 시행한 결과, 분리균주 1주는 Lactobacillus salivarius로, 다른 분리균주는 Lactobacillus delbrueckii subsp. lactis로 동정되었다. 7 분리균주를 16S rDNA partial sequencing으로 동정한 결과, 2주 모두 Lactobacillus salivarius subsp. salicinius와 유전자 유사치가 99.60%, 99.73%를 보여 Lactobacillus salivarius subsp. salicinius로 동정되었다. 이상의 결과를 종합하면 치아우식증이 없는 소아의 타액에서 분리된 유산균 중 과산화수소를 분비하여 인공치태 형성과 휘발성 유황화합물 생성을 억제하는 분리균주는 Lactobacillus salivarius subsp. salicinius로 동정되었다.

• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of the Pharmaceutical Society of Korea
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• pp.180.1-180.1
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• 2001

• 치위생과학회지
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• 제9권2호
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• pp.249-255
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• 2009

본 연구는 성인성 치주염 환자의 치은연하 치면세균막을 총 60개 치아에서 채취하여 F. nucleatum과 A. actinomycetemcomitans의 동정을 위해 세균배양법, single PCR법 및 mutliplex PCR법을 실시하였고, 세균 동정법간의 비교를 통해 다음과 같은 결과를 얻었다. 1. F. nucleatum과 A. actinomycetemcomitans의 동정을 위해 세균배양법, single PCR 및 multiplex PCR을 실시한 결과 F. nucleatum은 각각 12개(20.0%), 45개(75.0%), 43개(71.7%) 치아에서 양성반응을 보였지만, A. actinomycetemcomitans는 각각 0개(0.0%), 4개(6.7%), 1개(1.7%) 치아에서 양성반응이 나타났다. 2. F. nucleatum은 세균배양법에 비해 single PCR법 및 multiplex PCR법에서 높은 검출 빈도를 보여 좀 더 효율적인 세균 동정법으로 생각되었지만, 통계적으로는 유의한 차이가 없었다. 3. A. actinomycetemcomitans는 세균배양법에서 전혀 검출되지 않아 통계적으로 검정할 수 없었고, 세균 동정법간의 비교도 어려웠다. 4. F. nucleatum과 A. actinomycetemcomitans의 동정을 위한 single PCR법과 multiplex PCR법 간의 비교에서 두 세균 모두 검출 빈도에 있어서는 큰 차이를 보이지 않았지만, 통계적으로는 유의한 차이를 보였다.

• Restorative Dentistry and Endodontics
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• 제24권3호
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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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• 제40권2호
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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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• 제27권1호
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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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• 제25권2호
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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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• 제29권1호
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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.