다목적 배양기의 개발에 필요한 기초연구

양남웅 조선대학교 부설 의학연구소 2002 The Medical Journal of Chosun University Vol.27 No.2

Background and Objectives: An anaerobic chamber or jar for the anaerobic culture costs a great deal in upkeep, and most users are afraid of high-pressure gas tanks attached to a CO2 incubator or an anaerobic chamber. We devised the culture box with an air-bag and tested it to obtain the basic data for devising a multipurpose culture chamber which need not much upkeep expenses. Materials and Methods: For tests, we manufactured polymethyl methacrylate box (280 × 280 × 240 mm, in size and 18.8 L in volume) with a polyvinyl chloride air-bag. The CO2 concentration inside the box was measured by a CO2 analyzer and an aspirating pump set. The O2 concentration was measured by an oxygen indicator inside the box and the dissolved oxygen concentration in the media by the dissolved oxygen meter. Results: 5% CO2 was generated when 2.73 g of sodium bicarbonate and 6.83g of citric acid were mixed with 50 mL tap water inside the box. The absolute anaerobic condition was made within 45 minutes from mixing 3.5 g of sodium borohydride and 50 mL of 1 N acetic acid, and 5% O2 concentration (micro-aerobic condition) was produced within 15 minutes by mixing 20 mL of 1 N acetic acid and 1 g of sodium borohydride. Conclusion: The culture box with an air-bag needs neither CO2 and mixed gas tanks, nor an expensive and disposable anaerobic pack. I think these results could be used for devising a new multipurpose culture chamber.

사람 적혈구 용해액과 마혈청을 첨가한 Columbia agar base에서 Gardnerella vaginalis 임상 분리주의 약제 내성

양남웅,임용,심성희 대한감염학회 2003 Infection and Chemotherapy Vol.35 No.2

콜레라 장독소에 의한 털성장 촉진 현상의 기작에 관한 연구(1)

양남웅,신성희,임용,주세혁,정석진,김재수,박정평,기근홍,장정수 대한화학요법학회 1998 대한화학요법학회지 Vol.16 No.4

배경 : 콜레라 장독소로 vasopermeeability assay를 하던 중에 우연히 발견된 및 성장 촉진 효과를 보다 체계적으로 검증하고 작용 기작에 대하여 알아보기 위하여 실험하였다. 방법 : NZW rabbit, SD rat, F344 rat, ICR mouse, C57BL/6 mouse, Hairless mouse, hamster 각 3마리의 배부 털을 제거하고 콜레라 장독소를 파내 주사(5-10㎍) 또는 1회 도포하여 10일 후에 털 성장의 정도를 육안으로 그리고 micrometer로 관찰 측정하였다. 5㎍의 독소를 피내 주사한 5일 후에 SD rat의 조직을 절취하여 광학 현미경 및 전자현미경적 관찰을 하였다. 결과 : 콜레라 장독소를 피내 투여한 결과 토끼를 비롯해서 생쥐, 쥐, 햄스터에서도 털 성장촉진 현상을 보였으며 hairless mouse에서는 털성장 효과가 없었다. New Zealand White rabbit와 F344 rat의 배부 털을 제거하고 장독소 10㎍/0.1ml PBS을 피부에 도포한 결과, 피내 투여한 것과는 달리 고르지 않게 털의 성장 촉진 현상을 나타냈다. 털의 길이는 대조군에 비해서 현저히 증가하였으나 독소 투여량에 비례하지 않았으며, 털의 두께는 투여량에 비례하여 증가하였다. 털의 멸생도는 실험군과 대조군 사이에 유의한 차이가 없었다. 결론 : 콜레라 장독소는 털 성장 주기가운데 퇴조기와 휴지기에 영향을 미치지 않는 것으로 추정된다. 광학 현미경 및 전자 현미경의 소견에 따르면, 피내 투여된 콜레라 장독소는 모낭세포와 성장기의 모낭을 둘러싸고 있는 모세혈관 내피세포의 활성화를 통해 털 성장을 유도하는 것으로 보인다. 이러한 결과는 콜레라 장독소가 다양한 종류의 탈모증 치료에 사용될 가능성이 있음을 시사한다. Background : Cholera enterotoxin(CT) had the effect of hair growth acceleration in previous rabbit experiment. The aim of this study was to elucidate mechanism of CT action for hair growth acceleration. Methods : CT was administered to NZW rabbit, F344 rat. SD rat, ICR mouse. C57BL/6 mouse and Hairless mouse mtradermally. Hair length and Thickness mere measured by stereoscope and micrometer. Stereomicroscopic and transmit electron microscopic findings mere studied for CT effect. Results : Cholera enterotuxin(CT) has exhibited the accelerated hair growth effect also in rats znd mice as the same Kith rabbits, except with hairless mouse. When pasted on the hair removed skin areas of back mith CT 10 ㎍/0.1ml PBS in F344 strain of rat and New Zealand White rabbit, hair growth was irregular12 accelerated unlike intradermal injection of CT. Hair of experimental group WaS clearly longer than control group but not proportional to doses of toxin. Hair thichess was increased mith proportion to toxm doses. Density degree of hair growth was not meaningly different m the stereoscopic and histological findings between control and experimental group. Conclusion : It is presumed that CT has not activation effect to telogen phase of hair cycle. Judging by light microscopic and transmit election microscopic findings, it is suggested that intraderma1 CT injection may induce acceleration of hair growth through the activation of hair follicular cells and endothelia1 cells of microvasculature surrounding the anagen hair follicles.

Development and Evaluation of the Quick Anaero-system-A New Disposable Anaerobic Culture System

양남웅,김진만,최광주,장숙진 대한진단검사의학회 2010 Annals of Laboratory Medicine Vol.30 No.2

Background : We developed a new disposable anaerobic culture system, namely, the Quick anaero-system, for easy culturing of obligate anaerobes. Methods : Our system consists of 3 components: 1) new disposable anaerobic gas pack, 2) disposable culture-envelope and sealer, and 3) reusable stainless plate rack with mesh containing 10 g of palladium catalyst pellets. To evaluate the efficiency of our system, we used 12 anaerobic bacteria. We prepared 2 sets of ten-fold serial dilutions of the 12 anaerobes, and inoculated these samples on Luria-Bertani (LB) broth and LB blood agar plate (LB-BAP) (BD Diagnostic Systems, USA). Each set was incubated in the Quick anaero-system (DAS Tech, Korea) and BBL GasPak jar with BD GasPak EZ Anaerobe Container System (BD Diagnostic Systems) at 35-37℃ for 48 hr. The minimal inoculum size showing visible growth of 12 anaerobes when incubated in both the systems was compared. Results : The minimal inoculum size showing visible growth for 2 out of the 12 anaerobes in the LB broth and 9 out of the 12 anaerobes on LB-BAP was lower for the Quick anaero-system than in the BD GasPak EZ Anaerobe Container System. The mean time (±SD) required to achieve absolute anaerobic conditions of the Quick anaero-system was 17 min and 56 sec (±3 min and 25 sec). Conclusions : The Quick anaero-system is a simple and effective method of culturing obligate anaerobes, and its performance is superior to that of the BD GasPak EZ Anaerobe Container System. (Korean J Lab Med 2010;30:133-7)

사람 적혈구 용해액과 마혈청을 첨가한 Columbia agar base에서 Gardnerella vaginalis 임상 분리주의 약제 내성

양남웅,임용,신성희 대한감염학회 2003 감염과 화학요법 Vol.35 No.2

목적 : 국내에서 아직 보고가 없는 G. vaginalis에 대한 항균제 감수성 시험 결과를 얻기 위하여 실험하였다. 방법 : 악취가 나는 질 분비물의 증가를 주소로 내원한 여성 환자들에서 분리한 G. vaginalis 48주와 표준균주 1주(ATCC 14018)에 대하여 한천 희석법에 의한 항균제 감수성 시험을 하였다. 본 연구자들이 새로 고안한 방식으로 제조한 사람 적혈구 용해액을 마 혈청과 함께 columbia agar(1% proteose pepton No. 3함유)에 각 5%씩 첨가하여 한천 희석법의 시험 배지로 사용하였다. 결과 : Clidamycin과 ciprofloxacin의 MIC_(90)은 각각 0.3㎍/mL, 0.6㎍/mL로 감수성이 높았다. 기타 amoxicillin, doxycycline, erythromycin, cefazolin, gentamicin, metronidazole, tinidazole은 비교적 높은 내성을 보였다(NCCLS, U.S.A., 2001). 결론 : 세균성 질증의 치료를 위하여 metronidazole 혹은 tinidazole 단독 투여하기 보다는 clindamycin 또는 ciprofloxacin의 병용효과를 환자들에서 검증할 필요가 있다고 사료된다. 아울러 본 연구자들이 고안한 첨가물인 사람 적혈구 용해액은 G. vaginalis의 항균제 감수성 시험용 한천 희석 배지 및 시험관 희석 배지로 사용할 수 있을 것으로 생각된다. Background : To the best of our knowledge, there has been no any report on the antibiotic susceptibility profile of Gardnerella vagina/is, determined in domestic area by the agar dilution method. Therefore, we studied on 49 strains of G. vagina/is by the agar dilution method. Methods : One standard strain (ATCC 14018) and Forty-eight strains isolated from patients with increased vaginal discharge were included in this study. Columbia agar base containing 1% proteose peptone No. 3 was supplemented with horse serum (5%) and human erythrocyte lysate (5%) which was prepared by a new method, and this medium was used for the antibiotic susceptibility test. Results: The MICs_(90) of clindamycin and ciprofloxacin were 0.3 ㎍/mL and 0.6 ㎍/mL, respectively. Amoxicillin, cefazolin, doxycycline, and erythromycin were hardly effective against most strains of G. vagina/is (NCCLS, U.S.A., 2001). Especially, MICs_(90) of both metronidazole and tinidazole were 80 ㎍/mL under micro-aerobic condition of 5% O₂. Conclusion: For the treatment of Bacterial vaginosis, it is suggested that clindamycin or ciprofloxacin should be combined with vaginal tablet or gel of metronidazole rather than single administration of metrondazole or tinidazole.

Malassezia spp.의 최적 성장을 위한 Modified BHI Medium의 조성

양남웅 대한의진균학회 2015 대한의진균학회지 Vol.20 No.1

Background: There are several media for culture of Malassezia spp., such as Leeming & Notman(LN) medium, modified Leeming & Notman (mLN) medium, Dixon's medium and modified Dixon'smedium etc. It is known that Malassezia spp. grow well in these media in general, but the kind andamounts of their ingredients are various and un-uniform according to researchers. Author propose thenew and transparent BHI based medium for the optimal growth of Malassezia spp. Objectives: The purpose of this study was to design the simple and transparent BHI based mediumand find essential ingredients for the growth of M. globosa and M. obtusa. Methods: The colony size of eight standard strains (M. dermatis, M. furfur, M. globosa, M. japonica,M. obtusa, M. sloofiae, M. sympodialis, M. yamatoensis) on the modified BHI (mBHI) agar media withdifferent ingredients was observed by naked eye after seven day culture. The compositions of mBHImedium were as follows; mBHI-1 was supplemented with 0.7% dextrose, 1.5% Tween 80, 1% glycerolto BHI medium, mBHI-2 was supplemented with 1.5% Tween 40 to mBHI-1 instead of Tween 80,mBHI-3 was supplemented with 1.5% Tween 60 to mBHI-1 instead of Tween 80, mBHI-4 was addedwith 0.8% bile salts to mBHI-1. mBHI-5 was supplemented with 1.5% Tween 60 to mBHI-4 instead ofTween 80, and mBHI-6 was supplemented with 1.5% Tween 40 to mBHI-4 instead of Tween 80. pH ofsix mBHI media was all adjusted to 6.5. Results: M. furfur & M. japonica were grown well on mBHI-1 agar, but M. globosa & M. obtusawere not grown and others grown poorly. M. globosa & M. obtusa were not grown on mBHI-1 &mBHI-4 containing Tween 80 as lipid source, but others grown on all mBHI media. The media that alleight Malassezia strains grew well were slightly turbid mBHI-5 & transparent mBHI-6 medium. Conclusions: M. globosa & M. obtusa need glycerol and bile salts as well as Tween 60 or 40 insteadof Tween 80 for growth. M. furfur & M japonica need not bile salts for growth. Author proposes the transparent modified BHI medium supplemented with 0.7% dextrose, 1.5% Tween 40, 1% glycerol and0.8% bile salts (mBHI-6) as new standard medium for culture of eight Malassezia species.

Diagnosis of Bacterial Vaginosis with Relation to Isolation of Gardnerella vaginalis

양남웅,신성희,장정수 대한미생물학회 2002 Journal of Bacteriology and Virology Vol.32 No.1

환자, 병원 및 자연환경에서 분리된 녹농균의 감수성 분포와 혈청형

양남웅 中央醫學社 1989 中央醫學 Vol.54 No.3

장독소원성 대장균생산 내열성장독소의 장액분비자극에 대한 각종 약제의 억제효과

양남웅(Nam Ung Yang),박정평(Jung Pyong Park),이현국(Hyun Kook Rhee),주세혁(Se Hyuk Ju) 大韓微生物學會 1991 大韓微生物學會誌 Vol.26 No.3

Malassezia 효모와 Staphylococcus Aureus에 대한 제삼인산나트륨과 구연산의 탈ㆍ부착 효과와 성장억제 효과

양남웅 ( Nam-woong Yang ),정충현 ( Choong-hyun Jung ),이기완 ( Ki-wan Lee ),장숙진 ( Sook-jin Jang ) 한국환경기술학회 2012 한국환경기술학회지 Vol.13 No.3

이 연구는 제삼인산나트륨(trisodium phosphate, TSP)과 구연산(citric acid, CA) 및 그 병합제의 Malassezia furfur와 Staphylococcus aureus에 대한 탈-부착 (de-adherence) 효과와 성장억제효과를 평가하는데 목적이 있다. 이는 그것들을 건성비강진, 아토피피부염, 지루피부염과 같은 피부염 환자에 대한 보조요법제로 적용할 수 있을지를 결정하기 위한 것이다. M. furfur 에 대한 TSP 와 CA 일수소화물(monohydrate)의 탈-부착 효과와 성장억제 효과를 평가하는데 배지전개법(spreader plates method)을 사용하여 측정하였다. 아토피피부염을 악화시키는 것으로 알려진 S. aureus 에 대한 TSP 와 CA의 정균효과 (bacteriostatic effect)는 UV 분광광도계로 정량적으로 측정하였다. TSP 용액은 M. furfur 에 대해 탈-부착 효과를 나타냈다. 0.8 % TSP 를 함유한 Doubly modified Leeming and Notman (DmLN) 액체배지(pH 8.0)에서 M. furfur 균주들의 성장은 현저히 억제되었다. 다른 액체배지에 비해 0.4% CA를 함유한 brain heart infusion액체배지(pH 6.0)에서 S. aureus 는 27시간까지 뚜렷하게 억제되었다. 이 결과들은 TSP와 CA가 탈-부착 효과와 성장억제효과를 가지므로 건성비강진, 아토피피부염, 지루피부염과 같은 피부염에 보조요법제로 개발할 좋은 후보물질들이 될 수 있다는 것을 나타낸다. The aim of this study was to evaluate the de-adherence effect and growth inhibitory effect of the trisodium phosphate (TSP), citric acid (CA) and the combination of both agents on Malassezia furfur and Staphylococcus aureus to determine applicability of them as supportive care agents for patients with dermatitis such as dandruff, atopic dermatitis, and seborrheic dermatitis. The de-adherence and growth-inhibition effect of TSP and CA monohydrate on M. furfur were measured by spreader plates method. The bacteriostatic effect of TSP and CA on S. aureus, which is well known to exacerbate atopic dermatitis, was measured quantitatively by UV spectrophotometer. The TSP solution showed de-adherent effect on M. furfur. The growth of M. furfur strains were remarkably inhibited in Doubly modified Leeming and Notman (DmLN) broth containing 0.8 % trisodium phosphate (pH 8.0). S. aureus was strikingly inhibited until 27 hours in brain heart infusion broth containing 0.4% CA (pH 6.0), compared to other broths. The results indicate that TSP and CA, having de-adherent and growth inhibitory effect, could be good candidates to develop supportive care agents for dermatitis such as dandruff, atopic dermatitis, and seborrheic dermatitis.