배양 조건이 Candida albicans의 phospholipase 생성에 미치는 영향

신운섭,이경호,박주영,고춘명 대한의진균학회 1997 대한의진균학회지 Vol.2 No.2

Background : The dimorphic yeast, Candida albicans, is considered as a dangerous opportunistic pathogen in immunocompromised hosts. Several phospholipases of C. albicans are known to be secreted into the culture medium. Phospholipases have been proposed as a virulence factor in the pathoenesis of Candida infections. Objective : In order to investigate enzyme production, we examined culture condition of secreted phospholipase production from C. albicans. Methods : C. albicans ATTCC 10231 was cultivated in various media at 37℃ for 3 days. Phospholipase activity was measured by fatty acid soap precipitaiton in plate containing 0.04T lecithin, 0.1 M citrate buffer, pH 4.2 and 1.5% noble agar. Results : Phospholipase was highly induced when C. albicans was cultivated in broth medium(containing glucose 2%, albumin 0.2% and Fe^++ ion 0.01%) and Saboulaud's dextrose agar supplemented with 0.01% sodium deoxycholate. Conclusion : Highly induction of secreted phospholipase by albumin from C. albicans may be play an importans role in tissue invasion in the pathogenesis of C. albicans.

방선균 WCM-9가 생산하는 Polyene 항진균성 항생물질(AF1)의 생물학적 성상

신운섭,정선호,이동희,이경호,김수기,박주영,고춘명 대한의진균학회 1996 대한의진균학회지 Vol.1 No.1

Background: Pathogenic fungi infect humans, especially immunocompromised patients, with superficial or deeply invasive pattern. In the past 20 years, fungal infections have been increased dramatically resulted by increment of organ transplantation, cancer, AIDS patients, or use of broad-spectrum antibacterial agents. Fungal infections are now important causes of morbidity and mortality of hospitalized patients. but there is no effective antifungal antibiotics as well as antibacterial antibiotics Objective: Effective new antifungal antibiotics are needed for the treatment of mycosis. So in an effort to develop effective antifungal antibiotics, we screened over 600 isolates of Streptomyces spp. from soil. Methods: Antifungal producing strain was selected using disk diffusion method, An antifungal substance (AF1) was purified with ethyl acetate extraction, silica gel column chromatography and reverse phase HPLC. MICs of AF1 were detected by agar dilition method. Results: The compound showed UV maxima of 307, 321, 340, 359 nm indicating methylpentaene. Minimum inhibitory concentrations of the AF1 were 3.7 ㎍/ml against mold, and 3.7 - 7.4 ㎍/ml against Candida species. AF1 was also active against Cryptococcus neoformans, with MIC of 0.9 ㎍/ml. The concentration of AF1 for K^+ ion release from human red blood cell and hemolysis were 5 ㎍/ml. Conclusion: The antibiotic purified from culture broth of Streptomyces sp. WCM-9 was a polyene antifungal antibiotic which have broad spectrum antifungal activity.

캔디다균이 분비하는 단백질 분해효소의 순수 분리와 특성 및 교차 항원성

이경호,신운섭,박주영,김수기,박윤선,고춘명 연세대학교 원주의과대학 1995 원주의대논문집 Vol.8 No.1

Coxiella burnetii에 대한 항체의 검출을 위한 Enzyme-Linked Immunosorbent Assay의 정립

이경호,박주영,박윤선,김수기,신운섭,고춘명 대한감염학회 1994 감염 Vol.26 No.4

본 연구에서는 지금까지 진단이 내려지지 않았던 C. burnetii 감염증 환자들을 보다 빠르고 간편하며 정확하면서도 대량으로 검사할 수 있는 방법으로서 C. burnetii 단백항원을 이용한 효소면역측정법을 정립하고자 하였다. 본 실험에서 정립한 효소면역측정법은 간접 방법으로서 C. burnetii phase I에 대한 IgG를 검출하기 위해 고안되었다. C. burnetii 단백질 항원의 적정 농도는 야성 대조혈청의 흡광도와 음성 대조혈청에서의 흡광도의 차이가 가장 크게 나는 농도인 1ug/ml을 적정항원농도로 결정하였으며, 검사 혈청의 적정희석 농도도 역시 양성대조항원의 OD 값과 음성대조항원에서의 OD값의 차이가 가장 크게 나면서 비교적 background activity가 적은 혈청의 희석 농도인 1:300을 적정 혈청 희석 농도로 결정하였다. 이 때 각각의 야성 혈청이 0.185의 흡광도를 가질 수 있는 가장 높은 혈청 희석 농도의 로그값과, 같은 혈청을 1:300으로 희석했을 때의 흡광도 사이에 상관계수 r값이 0.95인 직선적 관계가 성립하여, 하가지 혈청 희석 농도(1:300)에서 결정된 ELISA값이 그 혈청의 항체가를 양적으로 표시하였다. 효소면역측정법의 양성 판정 기준은, IFA에서 C. burnetii에 대한 항체가 음성으로 확인된 혈청 100개를 대상으로 ELISA를 시행하여, 이들의 흡광도의 산술평균치와 표준편차를 구한 결과, 신뢰구간 95%에서 최고치인 0.185를 ELISA 야성 판정의 기준인 cut-off value로 정하였다. 위애서 정한 기준에 의해 ELISA를 시행하엿을 때, IFA에서 양성으로 판명된 혈청 모두에서 양성으로 나타나 민감도가 매우 높은 것으로 나타났다. 다음으로 ELISA 방법의 특이도를 조사하기 위해서, IFA 검사에서 음성으로 확인된 혈청(혈청 희석 농도 1:10에서 음성)을 대상으로 ELISA를 시행한 결과, blocking agent로서 1% BSA를 사용했을 때는 9.3%, 5% NGS는 11.3%, 그리고 blocking을 전혀 하지 않았을 때는 15.3%의 위양성율을 나타내어, BSA와 NGS 상이에는 통계학적으로 차이가 없었으나, blocking을 하지 않는 것 보다는 하는 것이 위양성율이 적은 것으로 나타났다. 그러나, 이것은 어디까지나 IFA기준이며, 절대적인 의미에서의 위양성율을 의미하는 것은 아니다. 결론적으로 본 연구에서 정립한 효소면역측정법은 매우 높은 민감도를 갖고 있으므로 Q fever가 의심되는 환자를 screening하기 위한 적절한 방법으로 사료된다. Currently, the indirect immunofluorescence assay(IFA) is used for detecting antibodies to Coxiella burnetii. Although reliable, this method is time-consuming, and the results are subject to interpretation. To develop an alternative test, we used an enzyme-linked immunosorbent asssy (ELISA) because it was faster, less complicated, and more objective than the IFA. The ELISA, unlike the IFA, can also be automated. In this study an indirect ELISA using C. burnetii protein antigens was developed to detect immunoglobulin G to C. burnetii phase I. A linear relation was found between the logarithms of absorbance values of sera at a dilution of 1/300 and the titres as determined by an end point dilution ELISA. The finding of linear relation (r=0.95) confirmed that an ELISA value determined with a single serum dilution (1:300) can be interpreted quantitatively as the titer of antibody in that serum. A serum dilution of 1:300 was used in all subsequent tests because at this dilution the difference between absorbance values for positive and negative sera was greatest: a serum sample was regarded as positive if the OD? value was ≥0.185 (absorbance threshold) at this dilution. The results obtained by the ELISA were compared with those by the IFA established already. The results indicate that the assay provides a sensitive, alternative method for diagnosing Q fever, but it needs reevaluation for the specificity because of its high false-positive rates.

Crossbar Technique for the Failed Clavicular Hook Plate Fixation in an Acute Acromioclavicular Joint Dislocation: Salvage for Acromial Fracture after Clavicular Hook Plate

Kyoung Hwan Koh,Dong Ju Shin,Seong Mun Hwang 대한견주관절의학회 2019 대한견주관절의학회지 Vol.22 No.3

We experienced acromial erosion and subsequent fracture after the treatment of Rockwood type V acromioclavicular dislocation with hook plate and coracoclavicular ligament augmentation. It was treated by using a surgical technique to address an acromial fracture and subsequent losses of reduction in acromioclavicular joint with two trans-acromial cortical screws (crossbar technique). The reduction state of acromioclavicular joint could be maintained by these two screws. Our crossbar technique could be considered as a good salvage procedure for the reduction loss caused by cutout or significant erosion of acromion after insertion of clavicular hook plate.

Ultrastructure of Fibrillar Layer of Candida albicans in Serum Culture

Shin, Woon-Seob,Kim, Dong-Hwa,Lee, Kyoung-Ho,Kim, Kyung-Hoon,Park, Yoon-Sun,Park, Joo-Young,Koh, Choon-Myung The Korean Society for Microbiology 2001 Journal of Bacteriology and Virology Vol.31 No.2

Adhesin, hydrolytic enzyme, and germ tube formation are regarded as major virulent factors of Candida albicans. In the pathogenesis of Candida albicans, the first step is adherence to host tissue with adhesin, and then invasion to the tissue by secreting hydrolytic enzymes like protease and phospholipase. The fuzzy coat is of interest as it may play important roles in antigenicity, phagocytosis, and adherence factors that are important in the virulence of C. albicans. The virulent factors could be expressed higher in vivo than in vitro and serum is similar to in vivo condition. Therefore, using transmitting electron microscopy, we studied the effects of serum on fibril expression of C. albicans YWMC132. The fibrillar layer of C. albicans was expressed only in serum The fibril expression was not inhibited at concentrations of C. albicans until $8{\times}10^7\;cells/ml$. However, the thickness of the fibril layer was reduced from $100{\sim}130\;nm$ to 20 nm according to increases in cell concentrations from $2{\times}10^7\;cells/ml$ to $8{\times}10^7\;cells/ml$. The fibrillar layer of C. albicans was expressed in serum, early, within 10 min, but the germ tube formation was expressed only after 60 min. When the fibrillar C. albicans was treated with lyticase, protease, or dithiothreitol (DTT), the fibrillar layer disappeared. The protein concentration was higher in the fibrillar cell than in the nonfibrillar cell. These results indicated that the proteinous fibrillar layer of C. albicans YWMC132 was expressed only in serum culture.

Oral Macrolide Therapy Following Short-term Combination Antibiotic Treatment of Mycobacterium massiliense Lung Disease

Koh, Won-Jung,Jeong, Byeong-Ho,Jeon, Kyeongman,Kim, Su-Young,Park, Kyoung Un,Park, Hye Yun,Huh, Hee Jae,Ki, Chang-Seok,Lee, Nam Yong,Lee, Seung-Heon,Kim, Chang Ki,Daley, Charles L.,Shin, Sung Jae,Kim, American College of Chest Physicians 2016 Chest Vol. No.

<P>CONCLUSIONS: Oral macrolide therapy after an initial 2-week course of combination antibiotics might be effective in most patients with M massiliense lung disease.</P>

A New Anthracene Based Fluorescent Turn-On Sensor for Fe3+

Kyoung-Lyong An,Sang-Oh Lee,Sun Duk Koh,Seung-Rim Shin,Jong-Il Shin,Soo Youl Park,손영아,박균하,Kun Jun 대한화학회 2014 Bulletin of the Korean Chemical Society Vol.35 No.1

Detection of Antibody to Candida albicans Proteinase in Sera from C. albicans-Cultured Patients and Healthy Controls

Kyoung-Ho Lee,Woon-Seob Shin,Hyun-Sook Park,Young Uh,In Ho Jang,Kap Jun Yoon,Choon-Myung Koh 大韓微生物學會 1997 大韓微生物學會誌 Vol.32 No.3

The Effect of Rabbit Serum on the Expression of Cell Surface Hydrophobicity in Candida albicans

Shin, Woon Seob,Kim, Dong Hwa,Lee, Kyoung Ho,Kim, Kyung Hoon,Park, Yoon Sun,Koh, Choon Myung 대한의진균학회 2000 대한의진균학회지 Vol.5 No.4

세포표면 소수성은 부착에 중요한 역할을 한다고 알려져 있으며 여러 가지 환경조건에 따라 세포표면 소수성의 표현이 달라진다. 그러나 혈청에 의해 유도되는 세포표면 소수성이 다른 조건에서 유도되는 세포표면 소수성과 같은지는 보고되지 않았으므로 혈청에 의한 칸디다균의 세포표면소수성을 관찰하였다. 토끼 혈청과 YEPD 배지에서 칸디다균을 37℃에서 배양하였을 때 초기 1시간 동안 세포표면 소수성이 급격하게 증가하다가 이후 서서히 감소하였다. 혈청을 희석하였을 때와 포도당을 첨가하여 배양하였을 때 칸디다균의 세포표면 소수성은 큰 차이가 없었으나 혈청을 많이 회석할수록 소수성이 높게 유지되었다. 칸디다균을 단백질분해효소와 lyticase로 처리하였을 때 칸디다균의 세포표면 소수성은 감소하였다. 칸디다균의 세포벽을 lyticase로 처리하여 전기영동으로 분석한 결과 단백질 양상이 소수성 균주간에도 모두 일치하지 않았다. 이러한 결과는 혈청에 의해 유도되는 세포표면 소수성과 YEPD 배지에서 유도되는 세포표면 소수성이 같지 않음을 의미한다. [Kor J Med Mycol 5(4): 167-172]