Polyphenolic compounds from Korean <i>Lonicera japonica</i> Thunb. induces apoptosis via AKT and caspase cascade activation in A549 cells

Park, Kwang Il,Park, Hyeonsoo,Nagappan, Arulkumar,Hong, Gyeong Eun,Yumnam, Silvia,Lee, Ho Jeong,Kim, Eun Hee,Lee, Won Sup,Shin, Sung Chul,Kim, Jin A,Lee, Sang Joon,Ma, Jin Yeul,Min, Taesun,Heo, Jeong D.A. Spandidos 2017 Oncology letters Vol.13 No.4

<P><I>Lonicera japonica</I> Thunb. (<I>L. japonica</I> T.) has historically been used in Korean herbal medicine due to its anticancer and protective effects on the respiratory system. In the present study, the polyphenolic compounds in <I>L. japonica</I> T. were investigated using high-performance liquid chromatography coupled with tandem mass spectrometry, and its anticancer effects on A549 non-small-cell lung cancer cells were studied. Polyphenolic compounds potentially inhibit A549 cells in a dose-dependent manner. Flow cytometry and western blot analysis demonstrated that polyphenolic compounds induce apoptosis by regulating the protein expression levels of caspases, poly-(ADP-ribose) polymerase and the B-cell lymphoma-2-associated X-protein/B-cell lymphoma-extra large ratio. Furthermore, polyphenolic compounds inhibited mitochondrial membrane potential activity. Caspase-3 activity was increased in a dose-dependent manner and polyphenolic compounds inhibited the activation of protein kinase B by dephosphorylation. These results suggest that polyphenolic compounds in A549 cells indicate the anticancer activity through the induction of apoptosis.</P>

도시관리계획에 따른 근린공원 차별화 방안 : 대구광역시 근린공원 대상으로

박정아(Jung-A Park),한상열(Sang-Yoel Han) 한국산림휴양학회 2010 한국산림휴양학회지 Vol.14 No.3

본 연구는 개발계획으로 생성된 근린공원 차별화방안에 관한 것이다. 대구광역시 근린공원의 대다수는 개발계획에 포함되어 생성되었는데, 각 공원은 각기 다른 용도지역에 위치해 있고, 용도지역별 이용자들의 수요 욕구도 다르다. 이에 공원이용자들을 대상으로 설문조사하고 이용행태를 분석하여 용도지역별 관리방안을 제시하였다. This study is about the differentiation plan of the neighborhood parks created in development plan. A large number of the neighborhood parks in Daegu Metropolitan City were laid out through the city development plan. Each of them is located in different areas where the park users have different needs. Hence, I intend to suggest a better way in laying out neighborhood parks based on the survey by the park users and the analysis about their behaviors when they use parks.

부산지역 유흥업소 종사여성으로부터 분리된 HPV16형의 발암유전자(E6/E7) 돌연변이 유형 분석

민상기(Sang-Kee Min),김성순(Sung Soon Kim),최병선(Byeong-Sun Choi),장대호(Dai-Ho Jang),이미옥(Mee-Ok Lee),최성화(Seung-Hwa Choi),김남호(Nam-Ho Kim),박연경(Yon-Koung Park),정영아(Yeong-A Jeong),김성준(Seong-Joon Kim),빈재훈(Jae-Hun Bi 한국생명과학회 2009 생명과학회지 Vol.19 No.6

HPV-16형의 염기배열 변이는 지역적, 인종적으로 특징적인 차이가 있으며 특히 HPV-16형 E6/E7 유전자의 특정 염기서열변이는 자궁경부암 및 자궁상피내 신생종양물의 발생을 일으키는 고위험 요인으로 알려져 있다. 본 연구는 2007년 부산지역 유흥업소 종사여성으로 분리된 HPV-16형 19건을 대상으로 E6/E7 유전자 영역(nt 34-880)을 표적으로 지역적 염기서열 변이를 조사하였다. nucleotide 수준에서 HPV16형 E6 유전자는 T178G (n=11), T178A (n=1), T350G (n=4), A442C (n=2), A104T, A111G, C116T, G145T, T183G, C335T, G522C 등 11종의 변이주가 발견되었고, E7 유전자는 A647G (n=12), A645C, A777C, G663A, T732C, T760C, A775T, T789C, T795G 등 9종의 변이주가 발견되었다. 아미노산 수준에서는 HPV-16형 E6 단백질의 경우 D25E (n=12), L83V (n=4), E113D (n=2), M1L, Q3R, P5S, Q14H, D25N, I27R, H78Y, C140S 등 11종의 변이주를, HPV16형 E7 단백질의 경우 N29S (n=12), L28F, T72S 등 3종의 변이주를 관찰할 수 있었다. 본 연구 결과, 부산지역의 HPV-16형 E6/E7 우점 돌연변이주는 E6 D25E (75%), E7 N29S (78%)로 각각 나타났다. 앞으로 자궁경부암 환자 및 일반여성을 포함한 더 많은 모집단을 대상으로 HPV-16형 E6/E7의 intratypic variants를 비교 조사하여 실제 HPV-16형 E6/E7 어떤 변이주가 자궁경부암 유발 위험성과의 관련성은 더 많이 연구되어져야 할 것으로 사료된다. Recent studies have reported that the distribution of HPV-16 sequence variation differs geographically, and more specifically that HPV-16 E6/E7 intratypic variants might carry a high risk for development of ICC (invasive cervical cancer) and CIN (cervical intraepithelial neoplasia) in a given population. To investigate the genetic diversities of HPV-16 E6/E7 oncogene by region, we collected nineteen HPV-16 isolates from sexually high-risk women in Busan, and analyzed the HPV-16 E6/E7 coding regions (nt 34 to 880) with HPV-16 E6/E7 specific PCR amplification. At the nucleotide level, eleven variants of the E6 genes and nine variants of the E7 genes were identified as follows: E6 T178G (n=11), E6 T178A (n=1), E6 T350G (n=3), E6 A442C (n=2), E6 A104T, E6 A111G, E6 C116T, E6 G145T, E6 T183G, E6 C335T, E6 G522C and E7 A647G (n=12), E7 A645C, E7 A777C, E7 G663A, E7 T732C, E7 T760C, E7 A775T, E7 T789C and E7 T795G, respectively. At the amino acid level, the isolated HPV-16 E6 and E7 genes showed eleven E6 variants: E6 D25E (n=12), E6 L83V (n=4), E6 E113D (n=2), E6 M1L, E6 Q3R, E6 P5S, E6 Q14H, E6 D25N, E6 I27R, E6 H78Y, E6 C140S and three E7 variants: N29S (n=12), L28F, T72S. HPV16 E6 L83V, the dominant variant in the Caucasian population, showed relatively low frequencies in our study population. We elucidated that the dominant HPV-16 E6/E7 variants were HPV-16 E6 D25E (63.2%) and HPV-16 E7 N29S (63.2%), which were phylogenetically included in Asian lineage. Further study is needed to evaluate the risk of cervical cancer related HPV-16 E6/E7 intratypic variants in the Korean population.

소셜 빅데이터를 활용한 도시공원 인식 비교 분석 연구: 양재시민의숲과 서서울호수공원을 대상으로

박상훈(Park, Sang-Hun),안영아(An, Young-A) 한국도시행정학회 2021 도시 행정 학보 Vol.34 No.4

이 연구는 온라인에서 발생하는 소셜 빅데이터 분석을 위해 유사한 주거지역에 위치한 두 개의 공원을 사례 대상으로 선정하여 비교분석 하였다. 이를 위해 도시공원에 대한 인식을 분석하고 개별공원의 특성과 이용상의 차이를 확인하는 방법으로써 소셜 네트워크 분석의 적용 가능성을 확인하고자 한다. 분석은 빅데이터를 근거로 하며, 연구의 핵심이 키워드 분석임에 따라 저자가 특허 등록한 ‘감성정보 분석 방법’의 일부를 활용하였다. 분석결과, 양재시민의 숲과 서서울호수공원은 계절을 체험하고, 느낄 수 있는 산책 공간으로 이용하고 있는 것으로 판단된다. 서서울호수공원은 주로 가족 단위로 공원의 시설과 공원 콘텐츠를 이용하는 반면에 양재시민의 숲은 가족 뿐만 아니라 친구 등 이용대상이 상대적으로 폭넓고 공원 시설 외에 양재천, 둘레길, 맛집 등 주변 환경과 연계된 이용 양상을 보이는 것으로 해석된다. 이러한 연구 결과는 도시공원 운영 및 관리에 있어 공원 이용객의 특성, 이용시간 등을 고려하여 프로그램을 개발하고 운영할 필요가 있음을 시사한다. In this study, two parks located in similar residential areas were selected as case subjects for the analysis of social big data analysis occurring online. For this purpose, the applicability of social network analysis is to be checked as a method of analyzing the perception of urban parks and identifying differences in characteristics and use of individual parks. The analysis is based on big data, and as the core of the study is keyword analysis, a part of the ‘emotional information analysis method’ registered by the author in the patent was used. As a result of the analysis, it is judged that Yangjae Citizen"s Forest and West Seoul Lake Park are being used as walking spaces where you can experience and feel the seasons. West Seoul Lake Park mainly uses the park"s facilities and park contents on a family basis. On the other hand, Yangjae Citizen"s Forest is interpreted as showing a relatively wide range of use for not only family but also friends, and it is used in connection with the surrounding environment such as Yangjaecheon Stream, Dullegil, and restaurants in addition to park facilities. These research results suggest that it is necessary to develop and operate programs in consideration of the characteristics and hours of use of park users in the operation and management of urban parks.

Oxidative lipid modification of nicastrin enhances amyloidogenic γ‐secretase activity in Alzheimer’s disease

Gwon, A‐,Ryeong,Park, Jong‐,Sung,Arumugam, Thiruma V.,Kwon, Yong‐,Kook,Chan, Sic L.,Kim, Seol‐,Hee,Baik, Sang‐,Ha,Yang, Sunghee,Yun, Young‐,Kwang,Choi, Yuri,Kim, Sa Blackwell Publishing Ltd 2012 AGING CELL Vol.11 No.4

<P><B>Summary</B></P><P>The cause of elevated level of amyloid β‐peptide (Aβ42) in common late‐onset sporadic [Alzheimer’s disease (AD)] has not been established. Here, we show that the membrane lipid peroxidation product 4‐hydroxynonenal (HNE) is associated with amyloid and neurodegenerative pathologies in AD and that it enhances γ‐secretase activity and Aβ42 production in neurons. The γ‐secretase substrate receptor, nicastrin, was found to be modified by HNE in cultured neurons and in brain specimens from patients with AD, in which HNE–nicastrin levels were found to be correlated with increased γ‐secretase activity and Aβ plaque burden. Furthermore, HNE modification of nicastrin enhanced its binding to the γ‐secretase substrate, amyloid precursor protein (APP) C99. In addition, the stimulation of γ‐secretase activity and Aβ42 production by HNE were blocked by an HNE‐scavenging histidine analog in a 3xTgAD mouse model of AD. These findings suggest a specific molecular mechanism by which oxidative stress increases Aβ42 production in AD and identify HNE as a novel therapeutic target upstream of the γ‐secretase cleavage of APP.</P>

수정-IPA를 통한 푸드카빙교육 효용성의 품질속성 분류에 관한 연구

이성주(Lee, Sung-Ju),임현철(Lim, Hyun-Cheo),김상미(Kim, Sang-Mi) 한국외식경영학회 2022 외식경영연구 Vol.25 No.2

본 연구는 푸드카빙 교육을 받은 수강생들이 지각하는 푸드카빙 효용성은 어떠한지 살펴보고, 어떠한 효용성들이 있고, 효용성의 품질 속성은 어떠한지 살펴보고자 하였다. 이러한 연구목적을 규명하기 위하여 푸드카빙 교육을 받은 경험이 있는 수강생을 모집단으로 설정하였고, 설문조사는 2020년 7월 20일부터 8월 20일까지 한 달간 진행되었다. 설문지는 총 500부를 배부하였고, 435부가 회수되었고, 불성실한 설문지 38부를 제외하고 397부가 최종 분석에 사용되었다. 분석결과, 중요 실행 요소로는 푸드카빙 교육은 지루하지 않고, 남녀노소 할 수 있고, 잡생각이 사라지고, 자존감이 높아지고, 자신의 가치가 높아지고, 마음이 편해지고, 우울함이 사라지는 등의 수업 자체에서 오는 만족감의 속성들이 포함되었다. 매력적 요소에는 푸드카빙 기술이 수익으로 연결되고, 강좌를 개설하고, 적은 금액으로 공방을 오픈하는 등의 수익 측면과 관련된 항목들이 포함되었다. 아울러, 기본적 요소에는 집중할 수 있고, 기분이 좋아지고, 자격증을 취득하고, 전문가가 될 수 있다는 측면의 항목들이 포함되었다. 이는 대체로 푸드카빙을 처음 배우는 수강생들이 기본적으로 가지는 목적들이 포함되었다. This study aims to examine the food carving utility perceived by students who received food carving education, the types of the utility, and its quality attributes. To investigate this research purpose, students who had received food carving education were set as a population, and the survey was conducted for one month from July 20 through August 20, 2020. 500 copies of the questionnaire were distributed in total, and 435 copies were collected. 397 copies were used in the final analysis, excluding 38 unreliable copies. As a result of the analysis, important execution factors included the attributes of satisfaction with the class itself, e.g., Food carving education was not boring; it could be done by men and women of all ages; it made cockeyed ideas disappear; it promoted their self-esteem; it enhanced their value; it comforted their hearts; and it made their depression disappear. Attractive elements included revenue-related items, such as food carving skills connected to profits; they could open lectures; and they could open a workshop with little money. Along with this, the basic elements included the following: They could concentrate; they could feel better; they could get a certificate; and they could become an expert. They usually included the purposes the students who would learn food carving for the first time had basically.

Biochanin A의 미백 효과에 관한 연구

박성하 ( Sung Ha Park ),김아현 ( A Hyun Kim ),박병준 ( Byoung Jun Park ),김진준 ( Jin Jun Kim ) 대한화장품학회 2013 대한화장품학회지 Vol.39 No.4

본 연구는 미백 소재 개발을 위하여 강진향(Dalbergia odorifera T. Chen) 추출물에서 분리한 활성물질인 biochanin A의 멜라닌 생성에 연관된 효과를 알아보았다. Biochanin A는 B16F1 melanoma 세포에서 농도의존적으로 멜라닌 양을 억제하였으며, 멜라닌 생합성 저해 효과는 10 μg/mL 농도에서 멜라닌 생성 억제율이 48%로 나타났다. Western blot을 이용하여 microphthalmia associated transcription factor (MITF), Tyrosinase, Trp-1와 Trp-2의 발현을 억제함을 확인하였다. 따라서 biochanin A는 미백 효능을 갖는 화장품 소재로서의 개발 가능성이 클 것으로 기대된다. To develop a new whitening agent for cosmetics, we investigated the inhibitory effects of biochanin A on melanogenesis. The biochanoin A is an active component in the extracts of Dalbergia odorifera T. Chen. It reduced melanin contents of B16F1 melanoma cells in a dose-dependent manner and decreased to about 48% at a concentration 10 μg/mL. It also inhibited the expression in microphthalmia associated transcription factor (MITF), Tyrosinase, Tyrosinase related protein 1 (Trp-1), and Tyrosinase related protein 2 (Trp-2) in melanocytes. The results suggested that biochanin A has considerable potential as a cosmetics ingredient with a whitening effect.

Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus

홍성욱,Kim Jong-Hui,Cha Hyun A,정건섭,Bae Hyo Ju,Park Won Seo,Ham Jun-Sang,Park Beom-Young,Oh Mi-Hwa 한국미생물·생명공학회 2022 Journal of microbiology and biotechnology Vol.32 No.11

Natural antimicrobial substances are needed as alternatives to synthetic antimicrobials to protect against foodborne pathogens. In this study, a bacteriocin-producing bacterium, Bacillus subtilis HD15, was isolated from doenjang, a traditional Korean fermented soybean paste. We sequenced the complete genome of B. subtilis HD15. This genome size was 4,173,431 bp with a G + C content of of 43.58%, 4,305 genes, and 4,222 protein-coding genes with predicted functions, including a subtilosin A gene cluster. The bacteriocin was purified by ammonium sulfate precipitation, Diethylaminoethanol-Sepharose chromatography, and Sephacryl gel filtration, with 12.4-fold purification and 26.2% yield, respectively. The purified protein had a molecular weight of 3.6 kDa. The N-terminal amino acid sequence showed the highest similarity to Bacillus subtilis 168 subtilosin A (78%) but only 68% similarity to B. tequilensis subtilosin proteins, indicating that the antimicrobial substance isolated from B. subtilis HD15 is a novel bacteriocin related to subtilosin A. The purified protein from B. subtilis HD15 exhibited high antimicrobial activity against Listeria monocytogenes and Bacillus cereus. It showed stable activity in the range 0–70°C and pH 2–10 and was completely inhibited by protease, proteinase K, and pronase E treatment, suggesting that it is a proteinaceous substance. These findings support the potential industrial applications of the novel bacteriocin purified from B. subtilis HD15.

Clinical Impact of Exosomal microRNA as a Novel Biomarker of Liver Fibrosis

( Young Chang ),( Jae-a Han ),( Suk Min Kang ),( Soung Won Jeong ),( Tom Ryu ),( Han Seul Park ),( Jeong-ju Yoo ),( Sae Hwan Lee ),( Sang Gyune Kim ),( Young Seok Kim ),( Hong Soo Kim ),( So Young Jin 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1

Aims: Many approaches have been suggested for the non-invasive diagnosis of liver fibrosis, including the use of serum biomarkers and ultrasound-based elastography, but none has yet replaced liver biopsy. MicroRNAs (miRNAs) have been suggested as potential diagnostic tools for liver diseases. We investigated alterations in the expression of serum exosomal miRNAs with the progression of liver fibrosis and evaluated their clinical applicability as biomarkers. Methods: This study prospectively enrolled 71 patients who underwent liver biopsy at a large-volume academic hospital in Korea. Exosomes were extracted from serum samples, and next-generation sequencing (NGS) of miRNAs was conducted in patients from different stages of liver fibrosis. Differential expression of miRNAs was quantified using targeted real-time quantitative polymerase chain reaction (RT-qPCR). A model was derived to discriminate advanced fibrosis based on miRNA levels using multivariate logistic regression. The performance of this model was evaluated and compared using area under the receiver operator characteristic (ROC) curve (AUC) and De- Long’s test. Results: NGS data revealed the relationship between exosomal miR-122 expression and liver fibrosis progression. The level of miR-122 decreased as the pathologic fibrosis grade progressed from stage 0 to 4. Patients with biopsy-proven advanced fibrosis had significantly lower levels of exosomal miR-122 (P<0.001) than those without advanced fibrosis. Exosomal miR-122 exhibited a fair performance in discriminating advanced fibrosis with an AUC of 0.77, which improved to 0.86 in combination with fibrosis-4 score (FIB-4) and transient elastography (TE). This value was higher than that reported for any other non-invasive modalities, including TE (AUC of 0.80) or FIB-4 (AUC of 0.57) alone. In a subgroup of patients with a non-viral etiology of liver disease, the performance of exosomal miR-122 as a biomarker improved, evident from the increase in the AUC value to 0.87. In this subpopulation, the combination model of miR- 122, FIB-4, and TE showed the best discrimination ability (AUC of 0.90), which was significantly higher than that of TE alone (AUC of 0.83; DeLong’s test P=0.046). Inhibition of miR-122 expression increased the proliferation of the human hepatic stellate cell line, LX-2, and upregulated the expression of collagen- 1A, a-smooth muscle actin, fibronectin, and transforming growth factor-ß. Conclusions: Exosomal miR-122 may serve as a novel biomarker for discriminating advanced liver fibrosis, and its accuracy may enhanced in combination with other non-invasive tests such as FIB-4 and TE.

경기도 식중독에서 분리된 Clostridium perfringens의 유전적 특성 분석

박성희,최옥경,정진아,김운호,이예은,박광희,윤미혜,Park, Sung-Hee,Choi, Ok-Kyung,Jeong, Jin-A,Kim, Woon-Ho,Lee, Yea-Eun,Park, Kwang-Hee,Yoon, Mi-Hye 한국미생물학회 2016 미생물학회지 Vol.52 No.3

질병관리본부에서 2012년 이후 수인성식품매개질환 실험실 진단실무지침에 cpb2 유전자를 포함시킨 후, 2013-2014년 경기도의 C. perfringens에 의한 식중독이 원인불명을 제외하고 가장 많이 발생되었으며, 그 발생률이 2011-2012년에 비해 큰 폭으로 증가하였다. 따라서 경기도내 유행하는 toxinotype을 파악하고, PFGE, MLST를 통해 이들의 분자역학적 연관성을 연구함으로서 C. perfringens에 의한 식중독 발생 시 역학조사 기초자료로 활용하고자 하였다. 2013-2014년 경기도에서 분리된 120주의 C. perfringens 중 cpe 보유균주는 49주, cpb2 보유균주는 71주였다. 발생건별로 살펴보면, cpe 단독발생건은 2건(10주), cpb2 단독발생건은 7건(45주), cpe, cpb2 혼합발생건은 7건(65주)로 cpb2 단독발생과 cpe, cpb2 혼합발생이 대부분을 차지하였다. Toxinotype PCR 결과, 120주 모두 C. perfringens에 의한 식중독의 주요 타입인 A로 밝혀졌다. PFGE 분석 결과, 53.5-100%의 유사도와 75 유형으로 나뉘었다. 65% 이상을 기준으로 했을 때 5가지 그룹으로 나뉘어졌다. A, C, D, E 그룹은 1개의 균주를 제외하고 모두 cpb2 보유균주로 이루어졌고, B 그룹은 1개의 균주를 제외하고 모두 cpe 보유균주로 구성되었다. 2014년 안산상록구 식중독 4주와 2014년 수원 권선구 식중독 3주를 제외하고, 64 cpb2 보유균주들은 대부분 다양한 유전자패턴을 보였다. 41 cpe 보유균주 중 3개의 균주를 제외하고, 2013년 부천원미구, 성남분당구, 2014년 안산상록구, 평택, 김포, 화성 식중독 모두 각각 동일한 유전자 패턴을 보였다. 2013 수원영통구 식중독은 2가지 cpe 유전형 패턴을 보였다. MLST 분석 결과, 크게 P-cpe 및 cpb2 그룹과 C-cpe 그룹으로 나뉘어 졌고, 세세하게 11개의 cluster로 나뉘어졌다. 경기도에서 분리된 31개 균주 중, 16 cpb2 보유균주와 2014-06-03 cpe 보유균주는 1-7 cluster에 속해있었고, 14 cpe 보유균주는 모두 8-11 cluster에 속해있었다. 하나의 cpe 보유균주를 포함하여 cpb2 보유균주들은 P-cpe 그룹과 건강한 사람에서 분리된 cpb2 그룹들 사이에 산재되어 cluster되었고, cpe 보유균주는 C-cpe 그룹에 속해있었다. 2014-06-03주의 cpe gene은 plasmid에 존재하고, 나머지 cpe 보유균주의 cpe gene은 모두 chromosome에 존재함을 추정 할 수 있었다. PFGE 및 MLST 분석 결과, cpe 보유균주에 비해 cpb2 보유균주가 훨씬 다양하고 복잡한 유전자패턴을 나타내며, cpe 유전자 보유균주의 경우 단일 유전자형이거나 유사도가 높은 유전자형으로 이루어져 있음을 알 수 있었다. cpe 보유균주의 경우 집단식중독의 원인균 파악이 용이하였으나, cpb2 보유균주의 경우 2 발생건을 제외하고 역학적인 연관성이 낮음을 확인 할 수 있었다. Clostridium perfringens is both a ubiquitous environmental bacterium and a major cause of human gastrointestinal disease, and C. perfringens food poisoning ranks among the most common gastrointestinal diseases in developed countries. 120 isolates of C. perfringens were obtained from food-poisoning outbreaks in 2013~2014, Gyeonggi-do. Using PCR, all 120 isolates were identified as C. perfringens type A. Of the tested isolates, 49 isolates carried the cpe gene, 71 isolates carried the cpb2 gene. The outbreak cases of cpb2 and cpe /cpb2 genes were 7 and 7, whereas the outbreak cases of cpe-gene were 2. The epidemiological relationship between C. perfringens isolates has previously been investigated chiefly by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). The genetics relatedness of the isolates raged from 53.5-100% and 75 district PFGE type were observed. The PFGE results revealed a wide genetic diversity among the 64 cpb2 carrying isolates (except 7 isolates), while 46 cpe-carrying isolates (except 3 isolates) showed a high genetic similarity. The MLST analysis revealed that 14 cpe isolates (cpe-chromosomal isolates) belong to a distinct cluster that is significantly distant from all the other cpb2 isolates (cpe-plasmid carrying and cpe-negative isolates). The isolates carrying a cpb2 appear to be rarely related, and are more variable than chromosomal cpe isolates. The results suggest that the cpe-positive outbreak isolates showed close genetic relation, whereas the cpb2-positive isolates revealed a wide genetic diversity.