급성 구강감염부위에서 Staphylococcus lugunensis 의 특성

유용욱,이미성,차정단,김기경,신상희,문상은,김강주 원광대학교 생명공학연구소 1999 생명공학연구소보 Vol.6 No.1

Staphylococcus lugdunensis (S. lugdunensis) is a newly identified pathogenic species of coagulase negative staphylococci and an occasional but not rare cause of severe infections, such as infective endocarditis after dental extraction, bacteremia, osteomyelitis, peritonitis, and soft tissue infections. As antibiotic use increased, resistance rapidly developed. Some strains have plasmids related to antibiotic resistance. To characterize S. lugdunensis in acute oral infection, S. lugdunensis was isolated from the patients with acute oral abscess, osteomyelitis, and normal persons. Antibiotic resistance, in vitro cellular toxicity, in vivo virulence, δ-like hemolysin activity, and synergistic hemolysis on sheep blood agar plates were investigated. The dot blot analysis and Southern blot analysis of staphylococcal DNA was performed with δ-hemolysin gene probe of Staphylococcus aureus. (S. aureus) Staphylococcal DNA was cloned, nucleotide sequence was analysed, and homology was compared with other sequence in Gene Bank. S. aureus, S. lugdunensis, Staphylococcus cohnii (S. cohnii), and other coagulase negative staphylococci (CNS) were isolated from the patients with acute oral infection. The isolation ratio of S. lugdunensis in the patients with infection was higher than that of healthy persons, but the isolation ratio of S. aureus in the patients with infection was similar to that of healthy persons. S. lugdunensis from the patients with acute oral infection showed the resistance to penicillin, methicillin, cephalothin, and clindamycin. S. lugdunensis in the patients had cellular toxicity in vitro and virulence in vivo. All strains of S. lugdunensis had δ-like hemolysin activity against rabbit erythrocytes. Four of the six strains of S. lugdunensis gave synergistic hemolysis with S. aureus on sheep blood agar plates. In dot blot analysis, all strains of S. lugdunensis showed the positive reaction with the probe of δ-hemolysin gene in S. aureus, but a 7.3 kb HindⅢ fragment was observed in the DNA of S. lugdunensis that gave synergistic hemolysis in a Southern blot analysis. The molecular size of partially purified δ-hemolysin was about 50 kd. The cloned fragments from the chromosomal DNA of S. lugdunensis showed the partial homology with the insulin receptor-related and dopamine receptor of humans. These results suggest that S. lugdunensis might be an important pathogen in acute oral infection and show some homology with eukaryotes.

소목(Caesalpinia sappan) 추출물의 Streptococcus mutans에 대한 성장, 산생성, 부착 및 비수용성 글루칸 합성 억제에 미치는 영향

유용욱,유현희,김윤정,유미선,서세정,이황,이흥수 대한구강보건학회 2003 大韓口腔保健學會誌 Vol.27 No.2

Streptococcus mutans is known as the causative bacterial playing the most important role in forming plaque and it is being noticed as major causative bacteria of dental caries. Therefore, development of more effective, substantial and safe preventive agent against dental caries and periodontal disease is honestly required. The present study was designed to investigate the effect of chloroform extract(CE), n-butanol extract(BE),methanol extract(ME) and water extract(WE) of Caesalpinia on the growth, acid production, adhesion, and insoluble glucan synthesis of Streptococcus mutans. The CE, BE, ME and WE decreased the growth and acid production of S.mutans than that of control, especially, the ME was more effective than other extracts. The effect of CE, BE, ME and WE on the adherences to saliva-coated hydroxyapatite bead(S-HA) were evaluated. At the concentration of 2,000 ㎍/㎖, the CE showed 32% rate of bacterial adherence to S-HA, whereas, other extracts of the same concentration producedgreater than 50% rate. The CE and BE were significantly decreased the reduction rates of water insoluble glucan formation from sucrose by crude glucosyltransferase of S.mutans. Thus, the application of C. sappan can be considered a useful and a practical method for the prevention of dental caries.

소목(Caesalpinia sappan) 추출물이 Streptococcus mutans의 치아우식 유발능 억제에 미치는 효과와 항균물질 brazilin의 분리

이황,유용욱 원광대학교 치의학연구소 2004 圓光齒醫學 Vol.13 No.1

Streptococcus mutans is known as the causative bacteria playing most important role in forming plaque and it is being noticed as the major causative bacteria of dental caries. Therefore, development of more substantially effective preventive agent against dental caries is required. "he present study was designed to investigate the effect of chloroform extract(CE), n-butanol extract(BE), methanol exmct(ME) and water extract(WE) of Caesalpinia sappan on the growth, acid production, adhesion, and insoluble glucan synthesis of S. mutans. Also, We investigated that chloroform, ethyl acetate, n-butanol and water fractions of C. sappan had antimicrobial activity against S. mutans and the most antibacterial activity shown component from ethyl acetate fraction bas been isolated and identified by LC/MS and NMR. All the exmcts decreased the growth and acid production of S. mutans than control, among which ME was the most effective. The effect of CE, BE, ME and WE on the adherence to saliva-coated hydmxyapatite bead(S-HA) was evaluated. At the concentration of 2,000 pgml, the CE showed 32% ratio of bacterial adherence to S-HA, whereas, other extracts of the same concentration were greater than 50% ratio. The CE and BE significantly decreased the formation of water insoluble glucan from sucme by crude glucosyltransferase of S. mutans. The ethyl acetate fraction from solvent partition showed greater antibacterial potential than other fractions. The compound was separated by preparative HPLC and was identified as Win by Lc/EsI/MS, 1H-NMR and 13C-NMR. The minimal inhibitory concentration(MIC) of brazilin against S. mutans was 63 pdml. These results implicated application of C. sappan as a useful agent for the prevention of dental caries.

구치부 복합레진의 중합 깊이에 따른 미세 경도에 관한 비교 연구

김은실,유용욱 원광대학교 치의학연구소 2003 圓光齒醫學 Vol.12 No.1

The introduction of many new posterior composite resins suggest that these products are rapidly gaining popularity. Curing efficacy is one of the important factor for the success of the composite resin restorations. The purpose of this study was to evaulate the microhardness and depth of cure and to determine if significant enhancements in physical and mechanical properties have been achieved for these materials compares with one popular conventional composite resins. For the three posterior and one conventional composites tested (SureFil, ALERT, Prodigy and Z100), the value for microhardness was determined. The depth of cure was evaluated by the measurements of microhardness. KHN (Knoop Hardness Number) values were then obtained at 0, 2, 3, 4, and 5 mm using a 100 g load and 10 s dwell time. Adequate depth of cure was determined as the bottom hardness being at least 80% of the top. The results were obtained as follows; 1. Conventional composite resin, Z100 showed harder surface than that of posterior composite resin. Upper surface was harder than lower surface. 2. The depth of cure of composite resins decreased with increasing thickness of resin. 3. No composite had adequate depth of cure when tested in increments greater than 2mm thick.

Berberine의 항생제 내성 황색포도구균에 대한 항미생물 효과

김해경,차정단,유용욱,김강주,이동근 원광대학교 생체재료·매식연구소 2000 원광생체재료·매식 Vol.9 No.1

Antibiotic-resistant Staphylococcus aureus (S. aureus) emerged in many parts of the world and increased complex clinical problems. Therefore, new agents are needed to treat to antibiotic-resistant S. aureus. Some natural products are candidates of new antibotic substances. Previous reports showed that the extract of Coptidis Rhizoma and Phellodendri Cortex have antimicrobial effects. The main antibacterial substance of Coptidis Rhizoma and Phellodendri Cortex is berberine. In this study, we examined the antimicrobial activity of berberine to antibiotic-resistant S. aureus. We isolated the 13 strains of antibiotic-resistant S. aureus from the oral cavity of healthy children in a preschool, and investigated the antimicrobial activity of berberine. We examined the effects of berberine on the level of resistance to antibiotic-resistant S. aureus. We observed that the invasion of S. aureus into human gingival fibroblasts was blocked by berberine. Berberine inhibited the growth of antibiotic-resistant. S. aureus as a dose dependent maner. Minimum inhibition concentration (MIC) of berberine against antibiotic-resistant S. aureus was 50-100 ㎍/ml. The inhibition of S. aureus invasion was depended on the concentration of berberine in human gingival fibroblasts. The MIC of ampicillin against antibiotic-resistant S. aureus was 32 ㎍/ml. However, the MIC of ampicillin against antibiotic-resitant S. aureus in the presence of 50 ㎍/ml berberine was 1 ㎍/ml. These data indicate that the combination with berberine and β-lactam antibiotics resulted in the synergistic reaction. These results suggest that the berberine may have the antimicrobial activities to antibiotic-resistant. S. aureus, and the mechanism may be the inhibition of growth and intracellular invasion. and alteration of antibiotic-resistance level of antibiotic-resistant S. aureus.

흰쥐 삼차신경 척수감각핵의 미측소핵 세포의 전압의존성 소디움통로와 포타슘통로에 대한 전기생리학적 연구

천상우,최정희,김권수,유용묵,김종구,이동근 대한악안면성형재건외과학회 2002 Maxillofacial Plastic Reconstructive Surgery Vol.24 No.1

The caudal subnucleus of the spinal trigeminal nucleus (medullary dorsal horn ; MDH) receives direct inputs from unmyelinated and thinly myelinated, small diameter primary afferent fibers. Thus, the MDH is implicated in the processing of nociceptive information in the orofacial region. In this study, the types and characteristics of voltage-dependent ion currents were investigated in acutely isolated MDH neurons of postnatal rats by means of the whole cell patch clamp techniques. Coronal slice (400㎛) of the trigeminal caudal subnucleus region was sequentially treated with pronase 0.2㎎/㎖. then single neurons were mechanically dissociated. Voltage-dependent sodium currents showed that the half-maximum activation potential was -41.8±1.8mV and half-maximum inactivation potential was -62.4±3.0mV. And the currents were blocked totally by application of 100nM tetrodotoxin. In a Ca^2+ free solution, low-threshold transient (I_A) and high-threshold sustained (I_K) currents were recorded. The half-maximum activation and inactivation potential of I_K were 2.5±1.9mV and -37.1±2.3mV, respectively. I_A was activated and inactivated more rapidly than I_K. The half-maximum activation and inactivation potential were -21.6±6.3mV and -84.5±5.0mV, respectively. When a 4-aminopyridine of 5mM was applied, I_A was almost totally blocked. These results reveal that MDH neurons express a variety of voltage-dependent ionic currents with distinct physiological and pharmacological properties, and they play an essential role in the transmission and modulation of sensation, especially pain, from trigeminal region.

Berberine의 항생제 내성 확색포도구균에 대한 항미생물 효과

김강주,차정단,김해경,유현희,유용욱,이동근 대한악안면성형재건외과학회 2002 Maxillofacial Plastic Reconstructive Surgery Vol.24 No.2

Antibiotic-resistant Staphylococcus aureus (S. aureus) emerged in many parts of the world and increased complex clinical problems. Therefore, new agents are needed to treat the antibiotic-resistant S. aureus. Some natural products are candidates of new antibiotic substances. Previous reports showed that the extract of Coptidis Rhizoma and Phellodendri Cortex have antimicrobial effects. The main antibacterial substance of Coptidis Rhizoma and Phellodendri Cortex is berberine. In this study, we examined the antimicrobial activity of berberine to antibiotic-resistant S. aureus. We isolated the 13 strains of antibiotic-resistant S. aureus from the oral cavity of healthy children in a preschool, and investigated the antimicrobial activity of berberine. We examined the effects of berberine on the level of resistance to antibiotic-resistant S. aureus. We observed that the invasion of S. aureus into human gingival fibroblasts was blocked by berberine. Berberine inhibited the growth of antibiotic-resistant S. aureus as a dosw dependent maner. Minimum inhibitory concentration (MIC) of berberine against antibiotic-resistant S. aureus was 50∼100㎍/㎖. The inhibition of S. aureus invasion was depended on the concentration of berberine in human gingival fibroblasts. The MIC of ampicillin against antibiotic-resistant S. aureus was 32㎍/㎖. However, the MIC of ampicillin against antibiotic-resistant S. aureus in the presence of 50㎍/㎖ berberine was 1㎍/㎖. These data indicate that the combination with berberine and β-lactam antibiotics resulted in the synergistic reaction. These results suggest that the berberine may have the antimicrobial activities to antibiotic-resistnat S. aureus, and the mechanism may be the inhibition of growth and intracellular invasion, and alteration of antibiotic-resistance level of antibiotic-resistant S. aureus.

산국 (Chrysanthemum boreale) 휘발성 추출액 및 EDTA가 미생물 및 인체 치은 섬유아세포에 미치는 영향

차정단,김태영,우원홍,전병훈,김해경,유용욱,김강주,길봉섭 원광대학교 생체재료·매식연구소 2000 원광생체재료·매식 Vol.9 No.1

Chrysanthemum boreale (C. boreale) has been used for the treatment of furuncles, carbuncles, sores, conjunctivitis, headache, vertigo and swollen throats as a folk medicine in Korea, but there is little report about experiment evidence of antimicrobial activity. To investigate the antifungal and antibacterial activity of Chrysanthemum boreale(C. boreale) volatile substance and EDTA, the growth inhibition against Aspergillus nidulans (A. nidulans), Fusarium oxysporum (F. oxysporum), Alternaria mali(Al. mali), Candida albicans (C. albicans), staphylococcus aureus (S. aureus), Streptococcus pyogenes(S. pyogenes), streptococcus mutans (S. mutans), Escherichia coli(E. coli), Helicobacter pylori (H. pylori) and Porphyromonas gingivalis (P. gingivalis) was monitored by agar dilution and broth microdilution method. The antifungal effect of C. boreale on A. nidulans, F. oxysporum, Al. mali and C. albicans was shown at the concentration of 0.04%. The antibaterial effect of C. boreale on S. aureus, S. pyogenes, S. mutans and H. pylori was shown at the concentration of 0.04%. However, the growth was inhibited at the concentration of 0.04% irrespective of the presence of EDTA 0.125% was much higher than higher than that of several concentration of volatile substance. The minimum inhibitory concentrations of volatile substance were 0.16% against S. aureus, 0.04% against S. pyogenes, 0.02% against S. mutans, 0.40% against E. coli, and 0.18% against C. albicans. The growth of human gingival fibroblasts was not affected by C. boreale volatile substances at the concentration of 0.04%. These results indicate that volatile substance from C. boreale may have biologically toxic activities to the microorganisms.

병원과 그 외부환경에서 분리된 포도상구균의 항균제 감수성과 카드뮴 저항성비교

김은숙,문상은,김강주,유용욱,정수진,김신무,문영희 원광대학교 생명공학연구소 1995 생명공학연구소보 Vol.3 No.1

It has been reported that Stratococcus aures(S. aureus) causes outbreaks of infection. To investigate the nosocomial infection, one hundred strains of S. aureus were isolated from the nasal cavity of the personnel and the environment in the dental clinic at Chonbuk province in Korea, the nasal cavity of personnel in healthy students, and the disease site from the patients at the Wonkwang medical center and Jesus presbyterian medical center. S. aureus was identified by Gram stain, catalase test, mannitol fermentation test and coagulase test. After isolation, the strains were characterized by antibiotic resistance. The antibiotic susceptibility tests were performed according to the National Committee of Clinical Laboratory Standards. Cadmium-resistance testing was assayed by broth dilution using Mueller-Hinton containing 125 μM of cadmium. Plasmid DNA was isolated by boiling method. All isolate of S. aureus in this study showed the resistance to ampicillin and penicillin, and the susceptibility to vancomycin. The resistance ratio of cadmium in the disease site was higher than that of the healthy site. These results suggest that S. aureus in the hospital might show the resistance to ampicillin and penicillin, and vancomycin be a drug of choice in the nosocomial infection.

Antibacterial Activity of curcuma longa against Methicillin-resistant Staphylococcus aureus

Yong Ouk You,Hyeon Hee Yu,Byung Hun Jeon,Seung Il Jeong,Jung Dan Cha,Shin Moo Kim,Kang Ju Kim 한의병리학회 2003 대한동의병리학회 학술대회논문집 Vol.2003 No.7