Close Encounter of V. vulnificus with Host Cells is a Prerequisite to the Cytotoxicity of yet Unidentified New Virulence Factor

Kim, Young-Ran,Kim, Choon-Mee,Rhee, Joon-Haeng The Korean Society for Microbiology 2003 Journal of Bacteriology and Virology Vol.33 No.1

Live Vibrio vulnificus is highly cytotoxic to host cells in vivo and in vitro. The two most representative cytotoxins, cytolytic hemolysin and elastolytic protease, have been regarded to play major roles in the cytotoxicity of V. vulnificus. To further determine roles of the two cytotoxins in V. vulnificus pathogenesis, we constructed a double mutant of vvhA and vvpE genes, encoding a hemolysin and a protease, respectively. However, the cytotoxicity and the $LD_{50}$ of a vvhA/vvpE double mutant showed no significant difference from those of the isogenic wild type strain. From these results, we came to speculate that yet unidentified, key cytotoxic factors should playa major role in the cytotoxic activity of V. vulnificus. The HeLa cells encountered with V. vulnificus became rounded, following detachment from the bottom of culture plate, and were killed eventually. However, the bacterial culture supernatant did not show any effect on the morphology and viability of HeLa cells. Also, no cytotoxicity could be noted when V. vulnificus was not allowed to contact with HeLa cells in the $Transwell^{(R)}$ system. Chloramphenicol, at lower concentration than minimum inhibitory concentration (MIC), decreased the cytotoxicity of a vvhA/vvpE double mutant to HeLa cells in a dose dependent manner. These results suggest that close encounter of V. vulnificus with host cells is a prerequisite to the cytotoxicity and that a yet unidentified virulence factor (s) should play an important role in the contact-dependent cytotoxicity.

Essential role of an adenylate cyclase in regulating <i>Vibrio vulnificus</i> virulence

Kim, Young Ran,Kim, Soo Young,Kim, Choon Mee,Lee, Shee Eun,Rhee, Joon Haeng Elsevier 2005 FEMS microbiology letters Vol.243 No.2

<P><B>Abstract</B></P><P><I>Vibrio vulnificus</I>, a halophilic estuarine bacterium, causes a fatal septicemia and necrotizing wound infection. To investigate the role of cAMP in <I>V. vulnificus</I> virulence regulation, an in-frame deletion mutant of the <I>cya</I> gene encoding adenylate cyclase was constructed. The <I>cya</I> null mutation resulted in a pleiotropic change of virulence phenotypes. The production of hemolysin and protease, the motility, and the cytotoxicity were decreased by the <I>cya</I> mutation. The defects in the <I>cya</I> mutant were functionally complemented <I>in trans</I> by a plasmid carrying the wild type <I>cya</I> allele. The <I>V. vulnificus cya</I> mutant exhibited a 100-fold increase in LD<SUB>50</SUB> to mice. The result indicates that cAMP plays an essential role in the global regulation of <I>V. vulnificus</I> virulence.</P>

Effect of Salinity, Temperature, and Glucose on the Production of Vibrio vulnificus Hemolysin

Kim, Hyun-Soo,Shin, Sung-Heui,Park, Hae-Ryoung,Lee, Shee-Eun,Kim, Choon-Mee,Kim, Soo-Young,Kim, Young-Ran,Lee, Hyun-Chul,Chung, Sun-Sik,Rhee, Joon-Haeng The Korean Society for Microbiology 2002 Journal of Bacteriology and Virology Vol.32 No.4

Among the exotoxins produced by V. vulnificus, hemolysin (HS) has been reported to be the most potent one. To investigate the factors up- or down-regulating HS production in the context of pathogenesis, we observed the effects of salinity or/and temperature shifting, glucose, and acidic pH on the production of HS by V. vulnificus C7184 strain in vitro. Significantly more HS was produced when V. vulnificus was cultured in 0.9% salinity and $37^{\circ}C$ than in 2.5% and $25^{\circ}C$. When the culture condition reflecting natural habitat of V. vulnificus (2.5% salinity and $25^{\circ}C$) was changed into that reflecting human body (0.9% salinity and $37^{\circ}C$), 2.5 fold or more HS was produced than in the V. vulnificus being cultured continuously in 0.9% NaCl at $37^{\circ}C$. This result suggests that V. vulnificus somehow recognizes the shifting in salinity and temperature and stimulate HS production. Glucose addition in the culture medium resulted in a dose-dependent decrease in the HS production. Glucose itself and acidic pH resulting from its metabolism both appeared to inhibit the HS production. Glucose in itself had more dominant role in suppressing the HS production than the lowered pH accompanying the metabolism of glucose. This result suggests that HS production is down-regulated in the presence of glucose and under environmental acidic pH.

Selection and characterization of tenascin C targeting peptide

Kim, Mee Young,Kim, Ok Ran,Choi, Yong Seok,Lee, Heuiran,Park, Keerang,Lee, Choon-Taek,Kang, Keon Wook,Jeong, Sunjoo Springer-Verlag 2012 Molecules and cells Vol.33 No.1

<i>Ex Vivo</i> Characteristics of Human Amniotic Membrane-Derived Stem Cells

Kim, Jiyoung,Kang, Hyun Mi,Kim, Haekwon,Kim, Mee Ran,Kwon, Hyuck Chan,Gye, Myung Chan,Kang, Sung Goo,Yang, H. Seung,You, Juice Mary Ann Liebert 2007 Cloning & stem cells Vol.9 No.4

<P>Cells were isolated from four human amniotic membranes, and their biological characteristics analyzed during ex vivo expansion. Morphologically homogenous populations of fibroblast-like cells were obtained from the second or third passage. Under the appropriate culture conditions, these human amniotic membrane-derived mesenchymal cells (HAM) were shown to differentiate into adipocytes, osteocytes, chondrocytes and neuronal cells, as visualized by Oil Red O, von Kossa, alcian blue, anti-Neu N, and anti-Gal C antibody staining, respectively. Immunophenotype analysis of HAM cells revealed the presence of antigens for SSEA-3, SSEA-4, collagen type-I, -II, -III, -IV, -XII, fibronectin, alpha-SMA, vimentin, desmin, cytokeratin18 (CK18), HCAM-1, fibroblast surface protein, and human leukocyte antigen (HLA) ABC. ICAM-1 protein was weakly detectable, and proteins of TRA-1-60, VCAM-1, von Willebrand factor, PECAM-1, and HLA DR were not detected. HAM cells reached senescence after 14.5+/-0.9 passages, over a period of 146.8+/-8.9 days, and underwent an average of 36.9 4.7 population doublings. RT-PCR analysis showed that all four HAM cell lines consistently expressed genes of Oct-4, Rex-1, SCF, NCAM, nestin, BMP-4, GATA-4, HNF-4alpha, vimentin, and CK18, regardless of the passage number. The genes of Brachyury, FGF-5, Pax-6, and BMP2 were never expressed. Strikingly, alpha-fetoprotein (alphaFP), HLA ABC, and HLA DR genes were expressed in an earlier passage but not expressed in later passages. Telomerase activity of two HAM lines was discernable upon the third passage. These observations strongly suggest that HAM might be immune-privileged and, thus, advantageous as therapeutic cells.</P>

Differentiating Uterine Sarcoma From Atypical Leiomyoma on Preoperative Magnetic Resonance Imaging Using Logistic Regression Classifier: Added Value of Diffusion-Weighted Imaging-Based Quantitative Parameters

Kim Hokun,Rha Sung Eun,Shin Yu Ri,Kim Eu Hyun,Park Soo Youn,Lee Su-Lim,Lee Ahwon,Kim Mee-Ran 대한영상의학회 2024 Korean Journal of Radiology Vol.25 No.1

Objective: To evaluate the added value of diffusion-weighted imaging (DWI)-based quantitative parameters to distinguish uterine sarcomas from atypical leiomyomas on preoperative magnetic resonance imaging (MRI). Materials and Methods: A total of 138 patients (age, 43.7 ± 10.3 years) with uterine sarcoma (n = 44) and atypical leiomyoma (n = 94) were retrospectively collected from four institutions. The cohort was randomly divided into training (84/138, 60.0%) and validation (54/138, 40.0%) sets. Two independent readers evaluated six qualitative MRI features and two DWI-based quantitative parameters for each index tumor. Multivariable logistic regression was used to identify the relevant qualitative MRI features. Diagnostic classifiers based on qualitative MRI features alone and in combination with DWI-based quantitative parameters were developed using a logistic regression algorithm. The diagnostic performance of the classifiers was evaluated using a cross-table analysis and calculation of the area under the receiver operating characteristic curve (AUC). Results: Mean apparent diffusion coefficient value of uterine sarcoma was lower than that of atypical leiomyoma (mean ± standard deviation, 0.94 ± 0.30 10-3 mm2/s vs. 1.23 ± 0.25 10-3 mm2/s; P < 0.001), and the relative contrast ratio was higher in the uterine sarcoma (8.16 ± 2.94 vs. 4.19 ± 2.66; P < 0.001). Selected qualitative MRI features included ill-defined margin (adjusted odds ratio [aOR], 17.9; 95% confidence interval [CI], 1.41–503, P = 0.040), intratumoral hemorrhage (aOR, 27.3; 95% CI, 3.74–596, P = 0.006), and absence of T2 dark area (aOR, 83.5; 95% CI, 12.4–1916, P < 0.001). The classifier that combined qualitative MRI features and DWI-based quantitative parameters showed significantly better performance than without DWI-based parameters in the validation set (AUC, 0.92 vs. 0.78; P < 0.001). Conclusion: The addition of DWI-based quantitative parameters to qualitative MRI features improved the diagnostic performance of the logistic regression classifier in differentiating uterine sarcomas from atypical leiomyomas on preoperative MRI.

Loss of heterozygosity of chromosome 3p in patients with carcinoma of the cervix

Kim, Jin Woo,Hong, Sung Ran,Park, So Yeon,Ryu, Hyun Mee,Park, Chong Taik,Lee, Ki Heon,Chung, Hwan Wook,Lim, Kyung Taek,Seong, Seok Ju,Park, In Sou,Shim, Jae Uk,Kim, Tae Jin 대한부인종양 콜포스코피학회 2002 Journal of Gynecologic Oncology Vol.13 No.1

청소년 리더십 검사도구의 타당화 및 리더십 특성 분석

김미숙(Mee Sook Kim),유효현(Hyo Hyun Yoo),박효정(Hyo Jung Park),전미란(Mi Ran Chun),박춘성(Choon Sung Park) 한국교육심리학회 2008 敎育心理硏究 Vol.22 No.1

교육기회의 메리토크라시 구조 ― 대학 유형과 서열별 진학기회를 중심으로 ―

김미란(Kim Mee-Ran) 고려대학교 교육문제연구소 2004 敎育問題硏究 Vol.0 No.20

본 연구는 한국의 대학생 조사를 통해 교육기회 구조를 실증적으로 분석한 것이다. 고등교육 기회에는 부모의 직업과 학력 등의 사회계층에 따른 불평등이 존재하며 특히 학업성적으로 좌우되는 대학 유형과 서열별 진학기회에도 부모의 사회계층요인이 작용하고 있음을 알 수 있었다. 이를 성별로 분석하면 고등교육 진학기회 뿐 아니라 대학 유형별·서열별 진학기회에 대한 출신 계층의 영향력은 남자보다 여자에게 강하게 나타난다. 특히 지방 출신의 여학생은 지방 출신의 남학생보다 서울 소재의 유명대학에 진학하는 기회가 적어 성에 따른 교육기회의 차별적 구조를 형성하고 있음을 알 수 있었다. 이 같은 분석을 통해 고등교육기회의 계층간 불평등 구조를 밝히고 교육기회의 성차에 대한 메카니즘을 시사했다는 점에서 중요한 의의를 찾을 수 있을 것이다. This paper analysis the structure of educational opportunities in South-Korea, Which has strong tradition of Confucianism and is famous for intense competition for admission to colleges and universities. In line with analysis of inquiry to college and university students in Seoul, it investigates how inequality in educational opportunities remains in South-Korean society in which the equal opportunities seem to be growing. The main findings are as follows : 1) in spite of popularization of higher education, there still remains inequality in educational opportunities due to social class of a student portrayed by job and educational background of his/her parents: and 2) the social class affects admission opportunities for college and university types ranked by the student's individual abilities in accordance with their school performance, the is, by merit. It is confirmed that inheritance of a mechanism reproducing the social class exists in the system of higher education of which opportunities are thought to be offered to everybody based on the meritocratic principle. However, 3) the Knowledge is gained that a factor of merit-that is, one's ability rather than that of the social class-dominates admission of male students the top universities in accordance with a mechanism that excellent male students from local towns and villages can enter top universities even though they belong to lower classes. Therefore, the result that a factor of the social class has strong effect on educational opportunities for female students is acquired. In spite of the fact that differences in educational opportunities and their correlation with the social class emerge from discrepancy between the two genders, mechanism how and why the differences that come out have been hardly studied. Therefore, the kind of previously overlooked viewpoint on female students considered in this paper may shed light on various aspects of gender studies in addition to studies of educational opportunities and social class.

Isocudraxanthone K Induces Growth Inhibition and Apoptosis in Oral Cancer Cells via Hypoxia Inducible Factor-1 <b><i>α</i></b>

Shin, Mee-Ran,Lee, Hwa-Jeong,Kang, Soo-Kyung,Auh, Q-Schick,Lee, Young-Man,Kim, Youn-Chul,Kim, Eun-Cheol Hindawi Publishing Corporation 2014 BioMed research international Vol.2014 No.-

<P>Isocudraxanthone K (IK) is a novel, natural compound from a methanol extract of the root bark of <I> Cudrania tricuspidata</I>. It has not been shown previously that IK possessed antitumor activity. We investigated the antitumor effects and molecular mechanism of IK and related signal transduction pathway(s) in oral squamous cell carcinoma cells (OSCCCs). The MTT assay revealed that IK had an antiproliferative effect on OSCCCs, in a dose- and time-dependent manner. IK induced apoptosis in OSCCCs, as identified by a cell-cycle analysis, annexin V-FITC and propidium iodide staining, and the nuclear morphology in cell death. IK caused time-dependent phosphorylation of Akt, p38, and ERK (extracellular signal-regulated kinase). In addition, IK increased the cytosolic to nuclear translocation of nuclear factor-<I>κ</I>B (NF-<I>κ</I>B) p65 and the degradation and phosphorylation of I<I>κ</I>B-<I>α</I> in HN4 and HN12 cells. Furthermore, IK treatment downregulated hypoxia-inducible factor 1<I>α</I> (HIF-1<I>α</I>) and its target gene, vascular endothelial growth factor (VEGF). Cobalt chloride (CoCl<SUB>2</SUB>), a HIF-1<I>α</I> activator, attenuated the IK-induced growth-inhibiting and apoptosis-inducing effects, and blocked IK-induced expression of apoptosis regulatory proteins, such as Bax, Bcl-2, caspase-3, caspase-8, and caspase-9, and cytochrome c. Collectively, these data provide the first evidence of antiproliferative and apoptosis-inducing effects of IK as a HIF-1<I>α</I> inhibitor and suggest it may be a drug candidate for chemotherapy against oral cancer.</P>