Immobilization of the Hyperthermophilic Archaeon Thermococcus onnurineus Using Amine-coated Silica Material for H₂ Production

배승섭,나정걸,이성목,강성균,이현숙,이정현,김태완,Bae, Seung Seob,Na, Jeong Geol,Lee, Sung-Mok,Kang, Sung Gyun,Lee, Hyun Sook,Lee, Jung-Hyun,Kim, Tae Wan The Korean Society for Microbiology and Biotechnol 2015 한국미생물·생명공학회지 Vol.43 No.3

초고온성 고세균 Thermococcus onnurineus NA1은 개미산, 일산화탄소, 또는 전분 등을 이용해서 수소를 생산하는 것으로 알려져 있다. 본 연구에서는 T. onnurineus NA1의 고정화 세포를 이용한 수소생산을 고찰하였다. 고정화 실험결과, T. onnurineus NA1은 표면에 아민기가 코팅된 규조토 담체에 정전기적 인력에 의해 효과적으로 고정화되었고, 1 g의 담체에 고정화 될 수 있는 최대 세포의 양은 71.7 mg-dcw로 확인되었다. 고정화 세포를 이용한 세 번의 반복회분식 배양을 통해 개미산으로부터 수소생산 특성을 고찰하였고, 그 결과 배양이 반복됨에 따라 고정화 세포 농도의 증가에 기인하여 초기수소생산속도가 2.3 에서 4.0 mmol l⁻¹ h⁻¹로 상당량 증가됨이 관찰되었다. 따라서, T. onnurineus NA1의 고정화세포 시스템은 수소생산을 위한 좋은 대안이 될 수 있을 것으로 사료된다. 본 연구는 초고온성 고세균의 고정화세포를 수소생산에 적용한 첫 번째 사례이다. Previously we reported that the hyperthermophilic archaeon, Thermococcus onnurineus NA1 is capable of producing hydrogen (H₂) from formate, CO or starch. In this study, we describe the immobilization of T. onnurineus NA1 as an alternative means of H₂ production. Amine-coated silica particles were effective in immobilizing T. onnurineus NA1 by electrostatic interaction, showing a maximum cell adsorption capacity of 71.7 mg-dried cells per g of particle. In three cycles of repeated-batch cultivation using sodium formate as the sole energy source, immobilized cells showed reproducible H₂ production with a considerable increase in the initial production rate from 2.3 to 4.0 mmol l⁻¹ h⁻¹, mainly due to the increase in the immobilized cell concentration as the batch culture was repeated. Thus, the immobilized-cell system of T. onnurineus NA1 was demonstrated to be feasible for H₂ production. This study is the first example of immobilized cells of hyperthermophilic archaea being used for the production of H₂.

치매간호중재 프로그램이 경증 치매노인의 Na, K, Ca, Cl, P, Cholesterol의 혈중농도 및 17-KS, 17-OHCS의 뇨배설량에 미치는 영향

나현주,조명숙,임욱빈 대한기초간호자연과학회 2002 Journal of korean biological nursing science Vol.4 No.2

This study was conducted for the evaluation of the effect of Dementia prevention nursing program on Urinary 17-KS, 17-OHCS, and Blood Na, Cl, K, Ca, P, cholesterol. The sample for the present study was composed of 16 demented elderly in D care center for the eldery in K city. The Dementia prevention nursing program consisted of concept memory training, music therapy, and art therapy. The program was performed twice a week and about ninety minutes was consumed for one session. The program had been administered for 8 weeks. We checked Urinary 17-KS, 17-OHCS, and Blood Na, Cl, K, Ca, P, cholesterol befor and after the program. Statistical analysis was performed by using descriptive statistics and Wlicoxon signed rank test of SAS system for window 6.12. The result were follows : 1. Urinary 17-KS 17-KS score increased significantly after intervention(p=0.005). 2. Urinary 17-OHCS 17-KS score increased significantly after intervention(p=0.00l). 3. Blood Na, Cl, K, Ca, P, cholesterol Na(p=0.0002), Cl(p=0.0001) K(p=0.0001), Ca(p=0.0028) decreased significantly after intervention. The results show that Dementia prevention nursing program increases Urinary 17-KS, the Dementia 17-OHCS, and decreases Blood Na, Cl, K, Ca, P, cholesterol. In conclusion, prevention nursing program can be used for the effective measure to decrease stress of the Demented elderly.

Cloning, Purification, and Characterization of a New DNA Polymerase from a Hyperthermophilic Archaeon, Thermococcus sp. NA1

( Yun Jae Kim ),( Hyun Sook Lee ),( Seung Seob Bae ),( Jeong Ho Jeon ),( Jae Kyu Lim ),( Yo Na Cho ),( Ki Hoon Nam ),( Sung Gyun Kang ),( Sang Jin Kim ),( Suk Take Kwon ),( Jung Hyun Lee ) 한국미생물생명공학회 2007 Journal of microbiology and biotechnology Vol.17 No.7

Original Articles : Effect of aldosterone on the amplification of oncolytic vaccinia virus in human cancer Lines

( Hyun Ju Lee ),( Ja Sung Rho ),( Shao Ran Gui ),( Mi Kyung Kim ),( Yu Kyoung Lee ),( Yeon Sook Lee ),( Jeong Eun Kim ),( Eu Na Cho ),( Mong Cho ),( Tae Ho Hwang ) 대한간학회 2011 Clinical and Molecular Hepatology(대한간학회지) Vol.17 No.3

Background/Aims: JX-594 is an oncolytic virus derived from the Wyeth vaccinia strain that causes replication-dependent cytolysis and antitumor immunity. Starting with a cross-examination of clinical-trial samples from advanced hepatocellular carcinoma patients having high levels of aldosterone and virus amplification in JX-594 treatment, we investigated the association between virus amplification and aldosterone in human cancer cell lines. Methods: Cell proliferation was determined by a cell-counting-kit-based colorimetric assay, and vaccinia virus quantitation was performed by quantitative polymerase chain reaction (qPCR) and a viral plaque assay. Also, the intracellular pH was measured using a pH-sensitive dye. Results: Simultaneous treatment with JX-594 and aldosterone significantly increased viral replication in A2780, PC-3, and HepG2 cell lines, but not in U2OS cell lines. Furthermore, the aldosterone treatment time altered the JX-594 replication according to the cell line. The JX-594 replication peaked after 48 and 24 hours of treatment in PC-3 and HepG2 cells, respectively. qPCR showed that JX-594 entry across the plasma membrane was increased, however, the changes are not significant by the treatment. This was inhibited by treatment with spironolactone (an aldosterone-receptor inhibitor). JX-594 entry was significantly decreased by treatment with EIPA [5-(N-ethyl-N-isopropyl)amiloride; a Na+/H+-exchange inhibitor], but aldosterone significantly restored JX-594 entry even in the presence of EIPA. Intracellular alkalization was observed after aldosterone treatment but was acidified by EIPA treatment. Conclusions: Aldosterone stimulates JX-594 amplification via increased virus entry by affecting the H+ gradient. (Korean J Hepatol 2011;17:213-219)

돌연변이에 의한 효모의 bur6 대립유전자 유도

나종길,유현숙 群山大學校基礎科學硏究所 1998 基礎科學硏究 Vol.13 No.-

The Ydr1/Bur6 heterodimeric complex is a general transcription represser in the yeast Saccharomyces cerevisiae. The BUR6 gene is an essential gene for cell viability and the counterpart of DRAP1 transcription corepressor in human. The BUR6 gene was mutagenized by in vivo mutagenesis using repair-deficient mutant XL1-red or in vitro mutagenesis using hydroxylamine. The mutagenized bur6 alleles replaced URA3 plasmid-borne BUR6 on 5-FOA medium by plasmid shuffle and screened for their pleiotropic phenotypes. Fifty seven bur6 alleles were isolated among 5,280 transformants. The fifty seven mutants included 17 FOA-sensitive mutants, 20 temperature-sensitive mutants (at 37℃) and 20 cold-sensitive mutants (at 17℃). We assume that the 17 FOA-sensitive mutants have non-functional bur6 alleles. We are conducting DNA sequence analysis to determine the nature of the bur6 mutations. These data will provide the basis of the structure-function of Bur6 protein. These bur6 alleles with conditional phenotypes will be used to generate revertants to look for suppressor genes of bur6 which might be involved in transcription.

폐렴구균 감염증에서 항생제 내성의 임상적 의미 : Asian Network for Surveillance of Resistant Pathogens(ANSORP) Study

정숙인,기현균,손준성,고관수,김나영,장현하,오원섭,백경란,이남용,김신우,이혁,--,송재훈 대한감염학회 2003 감염과 화학요법 Vol.35 No.5

목적 : 전 세계적으로 폐렴구균의 페니실린을 포함한 β-lactam과 마크로라이드 및 퀴놀론 계열의 항생제에 대한 내성이 급격히 증가하여 임상적으로 문제가 되고 있으나, 시험관내 내성이 실제 임상적 예후에 미치는 영향에 대해서는 아직까지 논란이 되고 있다. 본 연구는 폐렴구균의 항생제 내성률이 높은 아시아 지역에서 항생제 내성이 폐렴구균 감염증의 임상경과에 미치는 영향을 평가하기 위하여 시행되었다. 방법 : ANSORP에서 주관한 전향적 임상 연구로서 1999년 11월부터 2001년 8월까지 아시아 지역 11개국의 14개 기관에서 진단된 침습성 폐럼구균 감염증 환자를 대상으로 내성 균주 감염의 위험요인, 임상양상, 예후 등의 임상적 분석을 시행하였다. 결과 : 총 646명 환자의 평균연령은 30.1세(6일-89세)였고, 이전 3개월 내에 항생제 사용력이 있는 환자는 159명(32.4%)이었다. 질환별로는 폐렴이 377예(58.4%)로 가장 많았고, 중이염 67예, 뇌수막염 66예(10.2%), 일차성 균혈증 65예(10.1%)였다. 총 646균주 중 347균주(53.7%)가 페니실린 비감수성(중등도 내성 23.1%, 고도 내성 23.1%)이었고, 페니실린에 대한 MIC_(90)은 0.03㎍/mL에서 4㎍/mL까지의 분포를 보였다. 페니실린 비감수성 균주에 의한 감염증과 차이가 없었다. 페니실린 내성 폐렴구균 폐렴에 의한 사망률은 페니실린 감수성 폐렴구균 폐렴에 의한 사망률과 차이가 없었다(p=0.846). Erythromycin에 대한 고도 내성을 보이는 균주의 비율이 매우 높았으나, 실제 임상상이나 치사율에는 영향을 미치지 않았다(p=0.092). 페니실린 비감수성 폐렴구균에 의한 뇌수막염의 치사률도 감수성 균주에 의한 사망률과 차이가 없었다(p=0.059). 결론 : 본 연구의 결과 페니실린 및 베타 락탐 제제에 대한 시험관 내성은 폐렴구균 폐렴이나 뇌수막염의 임상상이나 치사율에 영향을 미치지 않는 것으로 확인되었다. 이는 내성의 정도와 연관될 것으로 생각되므로, 향후 고도 내성 균주가 증가할 때 추가 검증을 요한다. 아울러 macrolide 및 퀴놀론계 항생제 내성의 임상적 의미에 대한 후속 연구가 필요할 것으로 사료된다. Background : Despite the widespread emergence of antimicrobial resistance among pneumococcal strains worldwide, clinical implications of in vitro resistance still remain an open question. To evaluate the clinical impact of pneumococcal resistance in Asian countries where the prevalence of pneumococcal resistance was reported to be highest in the world, ANSORP has performed a prospective, multinational surveillance study with cases with invasive pneumococcal diseases in Asian countries. Methods : In vitro susceptibility of pneumococcal isolates was determined by broth microdilution tests with 16 antimicrobial agents. All enrolled cases of pneumococcal infections were analyzed with regard to demographic data, clinical features, risk factors and mortality. Results : A total of 646 patients with pneumococcal infections were enrolled from 14 centers in 12 countries between the period from November 1999 to August 2001. Pneumonia (58.4%) was the most common clinical disease followed by bacteremia (33.4%), otitis media (10.4%), and meningitis (10.2%). Among 646 isolates, 347 (53.7%) were penicillin non-susceptible (intermediate 23.1%, resistant 30.7%). MIG_(90)s for penicillin ranged from 0.03 (India) to 4.0 ㎍/mL (Korea, Taiwan, Vietnam, and Hong Kong). Overall mortality from pneumococcal diseases by penicillin non-susceptible strains was not different from that by susceptible strains. Pneumococcal pneumonia caused by penicillin- or erythromycin-resistant strains showed similar mortality, severity of illness, or complications to that by susceptible strains. Mortality from pneumococcal meningitis caused by penicillin non-susceptible strains was also similar to that by susceptible strains. Conclusion : Data suggest that current situation of in vitro resistance to penicillin or macrolides may not affect the mortality from pneumococal pneumonia or meningitis caused by antibiotic-resistant strains.

재관류하는 혈액의 조성 변화에 따른 부정맥의 양상

박금수,나흥식,남숙현 고려대학교 의과대학 1991 고려대 의대 잡지 Vol.28 No.1

To determine the cause of genesis of reperfusion arrhythmias, the left anterior descending coronary artery was cannulated and perfused by a carotid-coronary bypass wlth slde branch in 29 open-chest pentobarbital-anesthetized cats . Ischemia was produced by shunt occlusion during 20min. Thereafter the side branch was opened and the ischemic myocardium reperfused with unmodified arterial blood (13 cats). acidlc blood(pH 6.73-7.10, 8 cats), hypocalcemic blood(Ca^(++) 0.13-0.37mM, 3 cats), and venous blood(PO_2, 29.5-47.ImmHg, 3 cats). Each group was evaluated with respect to the incidence of ventricular premature beats, ventricular tachycardia, and ventricular fibrillation and the onset time of first arrhythmia of each arrhythmia in cats. The incidence of ventricular tachycardia was much lower in the acidic reperfusion group (three of 8 cats,38%) than in the unmodilfed reperfusion group(eleven of 13 cats. 85%), (p<0.O5) hypocalcemic reperfuslon group(three of 3 cats. 100%), and hypoxic reperfusion group(three of 3 cats, 100% ). And the incidence of ventricular fibrillation was much lower in the acidic reperfusion group(none of 8 cats, 0%) than in the unmodified reperfusion group(eleven of 13 cats, 85%),(p<0.O5) hypocalcemic reperfusion group(three of 3 cats, 100%), and hypoxic reperfusion group(two of 3 cats, 67%). The onset time of ventricular premature beat and ventricular tachycardia is later in acidic reperfusion group(158.9 ± 117.5sec., 70.8±54.70sec. (mean±S.E) than unmodified reperfusion group(6.78±1.29sec., 24.7±7.5sec. (mean±S.E.)). These results indicate that acidic reperfusion can prevent reperfusion-induced arrhythmias, presumably owing to a reduction of Ca^(++) influx into cells through Na^(++) -Ca^(++) exchange.

아시아 지역 폐렴 구균의 항균제 내성 현황 및 변화 양상 : Asian Network for Surveillance of Resistant Pathogens (ANSORP) Study

정숙인,김나영,손준성,기현균,고관수,서지연,장현하,김연숙,오원섭,백경란,이남용,송재훈,항생제 내성 감시를 위한 아시아 연합 (ANSORP) 대한감염학회 2004 감염과 화학요법 Vol.36 No.1

목적 : 1980년대부터 페니실린을 비롯한 β-lactam 계열과 비 β-lactam 계열 항균제에 대한 폐렴구균의 항생제 내성이 급격하게 증가하여 국내 뿐 아니라 세계적인 문제로 대두되고 있다. ANSORP 에서는 아시아 지역에서 폐렴구균의 항균제 내성과 그 변화 추이, 내성의 기전 및 위험인자를 분석하고자 본 국제 공동 연구를 시행하였다. 재료 및 방법 : 1999년 11월부터 2001년 8월까지 아시아 지역 12개국의 14개 기관에서 폐렴구균 감염증 환자의 임상 검체로 분리된 침습성 폐렴구균을 대상으로 액체배지 미량희석법을 이용하여 16개 항생제의 MIC를 구하였다. 위험인자의 분석을 위하여 폐렴구균 감염증 환자를 대상으로 의무기록지를 조사하였다. Quellung 반응을 통해 혈청형을 결정하였다. 결과 : 총 685균주 중 52.4%가 페니실린에 대하여 비감수성(중등도 내성 22.9%, 내성 29.5%)을 보였고, 그 빈도는 베트남(92.1%), 스리랑카(85.7%), 홍콩(67.9%), 한국(64.5%)의 순이었다. Erythromycin에 대한 비감수성 비율은 베트남(92.1%), 대만(87.7%), 한국(80.6%), 홍콩(76.8%) 등에서 높게 나타났고, 베트남, 대만, 한국, 홍콩 등의 erythromycin의 MIC_(90)이 32㎍/mL를 넘는 고도내성을 보였다. 홍콩, 싱가포르, 대만, 인도에서는 fluoroquinolone에 대한 내성 균주가 출현하였다. 전체 균주 중 다제내성 균주의 비율은 47.7%였다. 혈청형은 19형과 23형이 가장 많았고, 그 외 6형, 14형 등이 있었다. 페니실린 비감수성 감염증의 위험인자로는 5세 미만(OR=1.7; 95%, CI, 1.2-2.4; P=0.002), 기관지폐질환(OR=2.0; 95%, CI, 1.3-3.1; P=0.003), 악성종양(OR=2.3; 95%, CI, 1.2-4.6; P=0.015), 스테로이드의 사용(OR=2.8; 95%, CI, 1.1-7.4; P=0.032)이 확인되었다. 결론 : 침습성 폐렴구균 균주를 대상으로 한 본 연구의 결과 아시아 국가들에서 페니실린, β-lactam 및 macrolide에 대한 폐렴구균의 내성률 및 내성 정도가 급증하고 있음을 확인하였다. Background : Emergence of pneumococcal resistance became a global issue since 1990s. According to the ANSORP studies with clinical isolates and carriage isolates between 1996 and 1999, some Asian countries showed alarmingly high prevalence of resistance to penicillin and other antimicrobial agents. To investigate the changing trends of pneumococcal resistance, ANSORP study group has performed a multinational surveillance study with invasive pneumococcal isolates from Asian countries. Methods : All isolates from various invasive pneumococcal diseases were prospectively collected from 14 centers in 12 countries between November 1999 to August 2001. Broth microdilution tests with 16 antimicrobial agents were performed according to the NCCLS procedures. Serotyping was performed by means of Quelling reaction with use of group-specific antisera. Results : A total of 685 isolates were collected. Overall, 52.4% of invasive isolates from Asian countries were not susceptible to penicillin (intermediate (1), 22.9%, Resistant (R), 29.5%). Vietnam showed the highest prevalence of penicillin non-susceptibility (1 20.6%, R 71.4%) followed by Sri Lanka (1 71.4%, R 14.3%), Hong Kong (1 24.1%, R 76%) and Korea (1 9.7%, R 54.8%). China (I 19.8%, R 23.4%) and Malaysia (1 9.1%, R 29.5%) also showed remarkable increase in penicillin resistance compared with previous ANSORP data, which were less than 10%. Vietnam (92.1%), Taiwan (87.7%), Korea (80.6%), and Hong Kong (76.8%) showed high prevalence of erythromycin resistance. MIG_(90)S for ciprofloxacin were 4 ㎍/mL (Hong Kong) and 2 ㎍/mL (11 Asian countries except Hong Kong), respectively. Conclusion : Compared with previous data from ANSORP studies, antimicrobial resistance among invasive pneumococcal isolates has markedly increased in Vietnam, Sri Lanka, Taiwan, China, and Malaysia. Continuous surveillance of pneumococcal resistance in Asia is strongly warranted.

서울시내 고등학교 위탁급식의 급식비와 투자비의 실태 및 위탁업체의 기대수준 비교 분석

양일선,배환미,김현아,신서영,조미나,박수연,차진아,이보숙 한국식생활문화학회 2002 韓國食生活文化學會誌 Vol.17 No.5

The purposes of this study were to a) find out the operational characteristics of the contract-managed highschool foodservice in Seoul, b) investigate the expected level of meal-price and facilities investment cost perceived by contract-managed highschool foodservice managers c) compare the present level and expected level of meal-price and facilities investment cost. From October 12 to November 13 in 2001, the questionnaires were mailed to 249 high schools which was managed by contract foodservice company with respondent rate 40.2%. Data were analyzed using SPSS Win(10.0) for descriptive analysis and one group paired t-test. The results of this study were as follows ; 1. The student enrollment of highschools run by contract-managed foodservice was 1,243 with 72.6% participation rate of school lunch program. The average meal-price was 2,138 won. The average annual period of school foodservice operation was 156.78 days per year. The average contracting period was about 3 years. 2. The average cost concerned in the facilities investment amounts 169,578,180 won at the initial investment and 25,204,092 won at the repairs and maintenance cost in the course of operation. 3. The present level of meal-price and facilities investment cost were respectively 2,136won/meal and 171,157,336.72 won. And expected level.of meal-price and amount of facilities investment cost were 2,418.75 won and 121,353,215.19 won. Comparing the present level with expected level of the meal-price and facilities investment cost, expected level of meal-price was significantly higher than the present level of meal-price(p<.001) and expected level of facilities investment cost was significantly lower than present level of facilities investment cost(p<.001).

Enhancement of H₂ production by genetically engineering a hyperthermophilic archaeon Thermococcus onnurineus NA1

Min-Sik Kim(김민식),Seong Hyuk Lee(이성혁),AeRan Choi(최애란),Jeong-geol Na(나정걸),Tae Wan Kim(김태완),Hyun Sook Lee(이현숙),Sung Gyun Kang(강성균) 한국신재생에너지학회 2016 한국신재생에너지학회 학술대회논문집 Vol.2016 No.5