폐렴구균의 용혈능 변화와 pneumolysin 유전자의 변이 양상

오원섭,장현하,정숙인,김연숙,이혁,김신우,김성민,백경란,이남용,송재훈 대한화학요법학회 2002 대한화학요법학회지 Vol.20 No.3

목적 : 폐렴구균은 인체에서 폐렴, 수막염 및 패혈증 등을 일으키는 중요한 병원균으로 현재 전 세계적으로 항균제 내성이 큰 문제가 되고 있다. 하지만 폐렴구균의 항균제 내성에 따라 병독성이 어떻게 변화하는지에 대해서는 구체적으로 알려져 있지 않다. 이에 기능적 및 구조적 분석을 통하여 폐렴구균의 주요 병독성 인자인 pneumolysin의 병인론적 역할을 규명하고자 하였다. 방법 : 임상 검체에서 분리된 페니실링 감수성 균주 20주와 내성 균주 20주를 대상으로 하였다. Pneumolysin의 세포 독성의 변화를 관찰하기 위하여 각각의 균주들의 용혈능을 측정하였고, pneumolysin 유전자를 중합효소 연쇄반응으로 증폭한 후 증폭 산물의 유전자 구조를 분석하였다. 결과 : 페니실린 감수성 균주와 내성 균주간의 용혈능은 유의한 차이를 보이지 않았으나, 일부 용혈능이 매우 낮은 균주에서 Lys152→Thr, Thr-172→Ile, Lys-224→Arg와 같은 변이가 관찰되었다. 결론: pneumolysin의 세포 독성의 변화는 항균제 내성 정도와는 무관하였으나, 일부 용혈능이 저하된 균주에서 발견된 pneumolysin 유전자의 변이가 pneumolysin의 세포 독성에 영향을 주는 것으로 보인다. Background : Pnumolysin (Ply) is one of the most important virulence factors of Streptococcus pneumoniae. To investigate the functional and structural variation of Ply, hemolytic activity (HA) and gene sequences of pneumolvsin were determined. Methods : A total of 40 invasive pneumococcal isolated (20 penicillin-susceptible & 20-resistant strains) were analyzed. HA of each isolate was measured in crude extracts (1×10^8 CFU/mL) hourly from the time of inoculation to 9h. Crude extracts (50㎕) and DTT buffer (100㎕) were serially diluted in 96-well plate and mixed with 1% sheep blood (50㎕). HA was represented as the reciprocal of the greatest dilution, which resulted in the complete lysis. Two sets of oligonucleotide primers were used to amplify a 1,474-bp fragment containing the coding region and a 578-bp fragment of upstream region of ply gene, respectively. Amplified products were directly sequenced. Results : HA showed a wide variation from 0 to 87,480 regardiess of penicillin MICs, serotypes or specimen sources. Two bacteremic strains showed completely no hemolytic activity. No significant differences in HA between penicillin-susceptible (Mean± SE : 15,468± 4,693) and penicillin-resistant strains (21,384± 4,087) were found (p > 0.05).Sequencing of the coding region showed unique alterations in amino acids in strains with markedly reduced hemolytic activity (≤ 40): Thr-172-Ile and Lys-224-Arg. However, upstream region of ply gene was highly conserved. Conclusions : Ply activity was not related with antibiotic resistance. Absence of HA in some strains was associated with unique alterations in amino acid sequences in ply gene. Relationnship between genomic alteration and functional changes in pneumolysin should be further investigated.

Bloodstream Infections and Clinical Significance of Healthcare-associated Bacteremia: A Multicenter Surveillance Study in Korean Hospitals

Son, Jun Seong,Song, Jae-Hoon,Ko, Kwan Soo,Yeom, Joon Sup,Ki, Hyun Kyun,Kim, Shin-Woo,Chang, Hyun-Ha,Ryu, Seong Yeol,Kim, Yeon-Sook,Jung, Sook-In,Shin, Sang Yop,Oh, Hee Bok,Lee, Yeong Seon,Chung, Doo The Korean Academy of Medical Sciences 2010 JOURNAL OF KOREAN MEDICAL SCIENCE Vol.25 No.7

<P>Recent changes in healthcare systems have changed the epidemiologic paradigms in many infectious fields including bloodstream infection (BSI). We compared clinical characteristics of community-acquired (CA), hospital-acquired (HA), and healthcare-associated (HCA) BSI. We performed a prospective nationwide multicenter surveillance study from 9 university hospitals in Korea. Total 1,605 blood isolates were collected from 2006 to 2007, and 1,144 isolates were considered true pathogens. HA-BSI accounted for 48.8%, CA-BSI for 33.2%, and HCA-BSI for 18.0%. HA-BSI and HCA-BSI were more likely to have severe comorbidities. <I>Escherichia coli</I> was the most common isolate in CA-BSI (47.1%) and HCA-BSI (27.2%). In contrast, <I>Staphylococcus aureus</I> (15.2%), coagulase-negative <I>Staphylococcus</I> (15.1%) were the common isolates in HA-BSI. The rate of appropriate empiric antimicrobial therapy was the highest in CA-BSI (89.0%) followed by HCA-BSI (76.4%), and HA-BSI (75.0%). The 30-day mortality rate was the highest in HA-BSI (23.0%) followed by HCA-BSI (18.4%), and CA-BSI (10.2%). High Pitt score and inappropriate empirical antibiotic therapy were the independent risk factors for mortality by multivariate analysis. In conclusion, the present data suggest that clinical features, outcome, and microbiologic features of causative pathogens vary by origin of BSI. Especially, HCA-BSI shows unique clinical characteristics, which should be considered a distinct category for more appropriate antibiotic treatment.</P>

우이도의 사구 염생식물 분포와 현존량에 관한 연구

이점숙,이강세,임병선,김하송,이승호 군산대학교 기초과학연구소 2000 基礎科學硏究 Vol.15 No.-

The coastal vegetation was classified by the Braun-Blanguet's method and the standing biomass was investigated on the coastal areas of Uido at the period of June to November, 1999. The vegetation of the study areas were classified with 5 communities as follows: Isachaemum authephoroides community, Zosia macrostachya community, Calystegia soldanella community, Messerschmidia sibirica community, Vitex rotundifolia community. Standing biomass was highest in Isachaemum authephoroides community (564 g.d. w/㎡), followed by Vitex rotundifolia community (392 g.d. w/㎡) > Zosia macrostachya community (255 g.d. w/㎡) > Calystegia soldanella community (79 g.d. w/㎡) and Lowest in Messerschmidia sibirica community (68 g.d. w/㎡)

Proliferation Effect of Conditioned Medium Produced by Lymph Node Stromal Cells

Lee, Sang-Han,Moon, Sung-Hoon,Ha, Sang-Chul,Ma, Jin-Yeul,Kim, Min-Sook,Lee, Jin-Man,Kwon, O-Yu Korean Society of Life Science 2002 Journal of Life Science Vol.12 No.1

CS21 lymphoma cells that preferentially metastasize to lymph nodes after s.c. inoculation into BALB/c mice were grown in vitro in the presence of CA12 stromal cells isolated from lymph nodes. To obtain fundamental data on identification and characterization of the soluble factor(s) produced by CA12 stromal cells, we investigated the biological profile of the conditioned medium produced by CAl2 stromal cells. CAl2 conditioned medium has no affinity with Con A. CAl2 conditioned medium is associated with the proliferation of splenic T- and thymic T-cells with-out adding mitogen, although the conditioned medium cannot induce the differentiation of thymocytes. Additionally, we showed that H-7, not HA-1004 inhibits CS21 cell proliferation. These results suggest that CAl2 conditioned medium has a specific soluble factor(s) produced by lymph node stromal cells.

고흥 연안의 사구 염생식물 분포와 현존량에 관한 연구

이점숙,이강세,임병선,김하송,이승호 군산대학교 기초과학연구소 2000 基礎科學硏究 Vol.15 No.-

The coastal vegetation was classified by the Braun-Blanquet's method and the biomass was investigated at the wetland of Kohung. The vegetation of the study area was classified with 7 coastal communities as follows: Carex scabrifolia community, Suaeda maritime community, Carex kobomugi community, Tetragonia tetragonoides community, Zoysia sinica community, Vitex rotundifolia community and Phragmites communis community. Biomass was highest in Phragmites communis community(2,087g.d.w./㎡), followed by Zoysia sinica community(741g.d.w./㎡), Suaeda maritime community(535g.d.w./㎡), Vitex rotundifolia community(334g.d.w./㎡), Carex kobomugi community(267g.d.w./㎡), Carex scabrifolia community(236g.d.w./㎡) and lowest in Tetragonia tetragonoides community(195g.d.w./㎡)

한국인 집단의 FⅧ의 (CA)n STRP의 유전적 특성

이하규,이정숙 가톨릭대학교 자연과학연구소 1999 자연과학논문집 Vol.20 No.-

혈액 응고 '인자 Ⅷ(coagulation factor Ⅷ, FⅧ)은 혈액 응고 경로에 관여하는 인자로서 X 염색체에 존재하고, 이 인자의 손상에 의해 혈우병 A가 발병하게 된다. 본 연구에서는 FⅧ 유전자 내에 존재하는 다형적 부위 중 (CA)_(n) 초변이성다형(STRP)을 관찰함으로써 한국인 집단에서의 유전적 특성을 알아보고자 하였다. (CA)_(n) STRP는 대립유전자 21에서 29까지 다양하게 관찰되었으며, 대립유전자 23이 정점을 이루는 단일분포 양상(unimodal pattern)을 보였다. 이형접합율은 0.76, PIC(polymorphism information content)는 0.73으로 나타나 다형성이 높은 것을 나타났으며, 친자확인시 이용되는 POE(power of exclusion)는 0.91로 높게 나타났다. 혈우병 진단시의 유용성을 알아보기 위해 가계분석을 한 결과, 가계 진단에 유용한 것으로 나타났다. Coagulation factorⅧ(FⅧ) whose gene is located in X chromosome is involved in the coagulation pathway. A defect of this factor generates hemophilia A. To study the genetic characteristics of coagulation factor ⅧI in Korean population, (CA)_(n) STRP at FⅧ gene have been investigated. The repetition number of (CA)_(n) at intron 13 ranged from 21 to 29. Allele 23(39%) which was 23 repeats of (CA)_(n), are the most frequently observed in Korean population. Heterozygosities of (CA)n STR was 0.76. PIC(polymorphism information content) value calculated on (CA)n STR loci was 0.73. POE(power of exclusion) value used in paternity case was 0.91. Thus (CA)n STRP in FⅧ analysis can be used for not only hemophilia A diagnosis at pedigree analysis but also forensic science.

함평만 갯벌 조간대의 식생분포 및 현존량

이승호,이점숙,김종욱,김하송,임병선 木浦大學校 沿岸環境硏究所 1998 沿岸環境硏究 Vol.15 No.1

함평만 조간대 염생식물조사를 위하여 염생식물군락이 형성된 6개지역을 선정하여 식생분포 및 현존량을 조사하였다. 우점군락은 퉁퉁마디군락(Salicoenia herbacea community), 칠면초군락(Suaeda japonic community), 나문재군락(Suaeda asparagoides community), 좀보리사초군락(Carex pumilalini community), 가는갯능쟁이군락(Atriplex gmelini community), 갯개미취군락(Aster tripolium community), 갯잔디군락(Zoysia sinica community), 해홍나물군락(Suaeda maritima community), 갯질경군락(Limonium tetragonun community), 갈대군락(Phragmites communis community), 실망초군락(Erigeron bonariensis community)으로 구분되었다. 염생식물 군락의 생산량은 1,680∼2,170 g.d.w./㎡으로 갈대군락이 가장 높았고, 갯잔디 군락이 310∼500 g.d.w./㎡으로 가장 낮았다. The coastal vegetation was classified by the Braun-Blanquet's method and the biomass was investigated at the wetland of Hampyung-Man. The vegetation of the study area was classified with 11 coastal communities as follows: Salicornia herbacea community, Suaeda japonica community, Suaeda asparagoides community, Carex pumilalini community, Atriplex gmelini community, Aster tripolium community, Zoysia sinica community, Suaeda maritima community, Limonium tetragonun community, Phragmites communis community, Erigeron bonariensis community, Biomass was highest in Phragmites communis community (1,680∼2,170 g.d.w./㎡) and lowest in Zoysia sinica community (310∼500 g.d.w./㎡).

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.

RNase A 와 B 에 대한 Mouse Immunoglobulin의 생성 및 분리조건

전숙영,이광표,김하형 중앙대학교 약학연구소 1997 약학 논총 Vol.11 No.-

Ribonuclease A, B (RNase A, B) are the enzymes catalyse he hydrolysis of 3',5'-phosphodiester linkages of ribonucleic acids. RNase B(M.W. 15.5KDa) differs from RNase A(M.W. 13.6KDa) in that RNase B has five glycoforms consisting of Man_9GlcNAc_2, to Man_9GlcNAc_2 at the single glycosylation site (Asn-34). It has not been fully studied that the difference of glycosylation between RNase A and RNase B would make difference in production of polyclonal antibody when it was used as antigen. In the present study, male Balb/c mice aged 6-7weeks were used, and three mice per group were immunized intraperitoneally with 0.2ml of emulsion mixture containing 100ug RNase A or B in 0.01ml of PBS and 0.18ml of Freund's adjuvant, and immunized with PBS as a control. To identify the production of polyclonal antibody, total protein amount was detected by use of UV- spectrophotometer and tested 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). In order to know the affinity of polyclonal antibody to RNase A, B and to measure the concentration of polyclonal antibody. ELISA method had been applied using the mouse monoclonal antibody 6 isotypes (IgG1, IgG2a, IgG2b, IgG3, IgA, IgM) and monoclonal anti-goat/sheep IgG peroxidase. Protein A column was prepared to test the possibility of separation of the polyclonal antibody to each immunoglobulin class. As a result, the glycosylation between RNase A and B did not have an effect on production pattern of polyclonal antibody from ascites of mice immunized with RNase A or B. Second, in the present study, the polyclonal antibody made from mice immunized with RNase A or RNase B should be isolated to immunoglobulin class or subclass by use of various binding or elution buffer.

여성들의 자기효능감과 건강증진행위에 관한 연구

정숙희,하안례,심선녀,이용환,김광혁 고신대학교보건과학연구소 2003 보건과학연구소보 Vol.13 No.-

The results were as follows : The higher the degree of self-efficacy feeling, one of the recognition factor was the more did they do health-promoting activities (γ=0.48, p=0.00). The conducting degree of health-promotion activities resulted in 3.79 points in the perfect score of 6point, which was a quiet high result. The subordinate areas under the health-promoting activity were adjustment of personal relationshiP (4.42), nutrition (3.93), stress control (3.87), self-realization (3.64), alcoholic drink/smoking (3.38), and responsibility for health. As for the differences of self-efficacy feeling according to subjects general characters, significant differences were found in the areas of age, occupation, whether or not they have taken healthrelated lectures, family and geriatric diseases of family members or relatives. The above study results tell that the self-efficacy feeling has a great effect on their putting the health-promoting activities into practice, in daily life.