• Title/Summary/Keyword: CDC

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Purification and Characterization of Endotoxin from Vibrio vulnificus (비브리오 패혈증균의 균체내독소 정제 및 특성에 관하여)

  • 김영만;정현정;신일식
    • Journal of Life Science
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    • v.7 no.2
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    • pp.79-87
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    • 1997
  • To determine the cause of Vibrio septicemia by understanding the characteristics endotoxin from Vibrio vulnificus, lethal dose, heat resistance and vascular permeability enhancing activity were svaluated using vegestative cell and cell homogenate and the result is as follows: 1. Vibrio vulnificus CDC B3547 of patient origin did not exihibit any significant difference in toxicity compared to Vibrio vulnificus B57 of enviroment origin. 2. Strong toxicity was observed when viable cell count of Vibrio vulnificus CDC B3547 was more than 10$^{7}$/ml. 3. Toxicity of cell homogenate was completely inactivited upon geating at 80$^{\circ}$C for 20min. 4. Cell homogenate did not show hemolyic activity but was acknowleged to have cytotoxicity. 5. Major lethal toxin against mouse was existed in Vibrio vulnificus CDC B3547; however, separation of LPS and LPS-protein complex was not successful using the current technique.

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A Case-Study of Implementation of Improved Strategies for Prevention of Laboratory-acquired Brucellosis

  • Castrodale, Louisa J.;Raczniak, Gregory A.;Rudolph, Karen M.;Chikoyak, Lori;Cox, Russell S.;Franklin, Tricia L.;Traxler, Rita M.;Guerra, Marta
    • Safety and Health at Work
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    • v.6 no.4
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    • pp.353-356
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    • 2015
  • Background: In 2012, the Alaska Section of Epidemiology investigated personnel potentially exposed to a Brucella suis isolate as it transited through three laboratories. Methods: We summarize the first implementation of the United States Centers for Disease Control and Prevention 2013 revised recommendations for monitoring such exposures: (1) risk classification; (2) antimicrobial postexposure prophylaxis; (3) serologic monitoring; and (4) symptom surveillance. Results: Over 30 people were assessed for exposure and subsequently monitored for development of illness. No cases of laboratory-associated brucellosis occurred. Changes were made to gaps in laboratory biosafety practices that had been identified in the investigation. Conclusion: Achieving full compliance for the precise schedule of serologic monitoring was challenging and resource intensive for the laboratory performing testing. More refined exposure assessments could inform decision making for follow-up to maximize likelihood of detecting persons at risk while not overtaxing resources.

Implementation of Polarization-Insensitive Directional Coupler using Curved Waveguides (곡면형 도파로를 사용한 편광 무의존성 방향성 결합기의 구현)

  • Ho, Kwang-Chun
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.16 no.1
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    • pp.239-244
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    • 2016
  • The polarization characteristics of polarization-insensitive directional coupler based on double sandwiched rib-type and curved waveguides are explored in detail by using conformal transformation method (CTM) and longitudinal modal transmission-line theory(L-MTLT). To obtain the polarization-insensitive condition of polarization-insensitive curved directional coupler(PI-CDC), the coupling length and coupling efficiency according to the inner radius of PI-CDC are analyzed for quasi-TE and quasi-TM modes. The numerical results show that the PI-CDC with a few micrometer scales can be realized by properly choosing the curvature and structural and material parameters of double sandwiched layers. Furthermore, the mode profiles propagating through PI-CDC are evaluated, and the influence on coupler performance has been investigated.

Direct characterization of E2-dependent target specificity and processivity using an artificial p27-linker-E2 ubiquitination system

  • Ryu, Kyoung-Seok;Choi, Yun-Seok;Ko, Jun-Sang;Kim, Seong-Ock;Kim, Hyun-Jung;Cheong, Hae-Kap;Jeon, Young-Ho;Choi, Byong-Seok;Cheong, Chae-Joon
    • BMB Reports
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    • v.41 no.12
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    • pp.852-857
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    • 2008
  • Little attention has been paid to the specificity between E2 and the target protein during ubiquitination, although RING-E3 induces a potential intra-molecular reaction by mediating the direct transfer of ubiquitin from E2 to the target protein. We have constructed artificial E2 fusion proteins in which a target protein (p27) is tethered to one of six E2s via a flexible linker. Interestingly, only three E2s (UbcH5b, hHR6b, and Cdc34) are able to ubiquitinate p27 via an intra-molecular reaction in this system. Although the first ubiquitination of p27 (p27-Ub) by Cdc34 is less efficient than that of UbcH5b and hHR6b, the additional ubiquitin attachment to p27-Ub by Cdc34 is highly efficient. The E2 core of Cdc34 provides specificity to p27, and the residues 184-196 are required for possessive ubiquitination by Cdc34. We demonstrate direct E2 specificity for p27 and also show that differential ubiquitin linkages can be dependent on E2 alone.

Naphthoquinone Analog-induced G1 Arrest is Mediated by cdc25A Inhibition and p53-independent p21 Induction in Human Hepatocarcinoma Cells

  • Kim, Won-Ho;Kim, Jung-Woong;Jang, Sang-Min;Song, Ki-Hyun;Ham, Seung-Wook;Choi, Kyung-Hee
    • Animal cells and systems
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    • v.11 no.1
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    • pp.9-15
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    • 2007
  • The naphthoquinone analog (2,3-dichloro-6,9-dihydroxy-1,4-naphtoquinone, NA) has an inhibitory effect on cdc25A protein phosphatase in vitro, which is responsible for G1/S transition during cell cycle. However, the exact mechanism inducing the growth inhibition is not understood. In this study, we investigated the regulatory mechanisms of growth arrest induced by NA, as a new potent inhibitor of cdc25A phosphatase, in human hepatocarcinoma SK-hep-1 cells. We found that NA induced the G1 arrest by perturbation of protein tyrosine dephosphorylation of Cdk2, which may be resulting from inhibition of cdc25A phosphatase. In addition, p21 was expressed in a p53-independent manner and participated in the NA-induced G1 arrest by inhibiting Cdk2 activity. Although the exact mechanism is not known, the p21 expression might be related to MAPK activation. From these results, we suggest that NA induces G1 arrest via inhibition of cdc25A and induction of p53-independent p21 expression in SK-Hep-1 cells.

Ginsenoside 20(S)-Rg3 reduces KIF20A expression and promotes CDC25A proteasomal degradation in epithelial ovarian cancer

  • Rong Zhang;Lei Li;Huihui Li;Hansong Bai;Yuping Suo;Ju Cui;Yingmei Wang
    • Journal of Ginseng Research
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    • v.48 no.1
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    • pp.40-51
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    • 2024
  • Background: Ginsenoside 20(S)-Rg3 shows promising tumor-suppressive effects in ovarian cancer via inhibiting NF-kB signaling. This study aimed to explore the downstream tumor suppressive mechanisms of ginsenoside Rg3 via this signaling pathway. Materials and methods: A systematical screening was applied to examine the expression profile of 41 kinesin family member genes in ovarian cancer. The regulatory effect of ginsenoside Rg3 on KIF20A expression was studied. In addition, we explored interacting proteins of KIF20A and their molecular regulations in ovarian cancer. RNA-seq data from The Cancer Genome Atlas (TCGA) was used for bioinformatic analysis. Epithelial ovarian cancer cell lines SKOV3 and A2780 were used as in vitro and in vivo cell models. Commercial human ovarian cancer tissue arrays were used for immunohistochemistry staining. Results: KIF20A is a biomarker of poor prognosis among the kinesin genes. It promotes ovarian cancer cell growth in vitro and in vivo. Ginsenoside Rg3 can suppress the transcription of KIF20A. GST pull-down and co-immunoprecipitation (IP) assays confirmed that KIF20A physically interacts with BTRC (β-TrCP1), a substrate recognition subunit for SCFβ-TrCP E3 ubiquitin ligase. In vitro ubiquitination and cycloheximide (CHX) chase assays showed that via interacting with BTRC, KIF20A reduces BTRC-mediated CDC25A poly-ubiquitination and enhances its stability. Ginsenoside Rg3 treatment partly abrogates KIF20A overexpression-induced CDC25A upregulation. Conclusion: This study revealed a novel anti-tumor mechanism of ginsenoside Rg3. It can inhibit KIF20A transcription and promote CDC25A proteasomal degradation in epithelial ovarian cancer.

A Generic Time-resolved Fluorescence Assay for Serine/threonine Kinase Activity: Application to Cdc7/Dbf4

  • Xu, Kui;Stern, Alvin S.;Levin, Wayne;Chua, Anne;Vassilev, Lyubomir T.
    • BMB Reports
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    • v.36 no.4
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    • pp.421-425
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    • 2003
  • The serine/threonine protein kinase family is a large and diverse group of enzymes that are involved in the regulation of multiple cellular pathways. Elevated kinase activity has been implicated in many diseases and frequently targeted for the development of pharmacological inhibitors. Therefore, non-radioactive antibody-based kinase assays that allow high throughput screening of compound libraries have been developed. However, they require a generation of antibodies against the phosphorylated form of a specific substrate. We report here a time-resolved fluorescence assay platform that utilizes a commercially-available generic anti-phosphothreonine antibody and permits assaying kinases that are able to phosporylate threonin residues on protein substrates. Using this approach, we developed an assay for Cdc7/Dbf4 kinase activity, determined the $K_m$ for ATP, and identified rottlerin as a non-ATP competitive inhibitor of this enzyme.

Molecular Docking Analysis of Protein Phosphatase 1D (PPM1D) Receptor with SL-175, SL-176 and CDC5L

  • Madhavan, Thirumurthy
    • Journal of Integrative Natural Science
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    • v.11 no.1
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    • pp.25-29
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    • 2018
  • Protein phosphatase manganese dependent 1D (PPM1D), a Ser/Thr protein phosphatise, play major role in the cancer tumorigenesis of various tumors including neuroblastoma, pancreatic adenocarcinoma, medulloblastoma, breast cancer, prostate cancer and ovarian cancer. Hence, analysis on the structural features required for the formation of PPM1D-inhibitor complex becomes essential. In this study, we have performed molecular docking of SL-175 and -176 and protein-protein docking of CDC5L with PPM1D. On analysing the docked complexes, we have identified the important residues involved in the formation of protein-ligand complex. Research concentrating on these residues could be helpful in understanding the pathophysiology of various tumors related to PPM1D.

Growth Arrest by Bufonis Venenum is Associated with Inhibition of Cdc2 and Cdc25C, and Induction of p21WAF1/CIP1 in T24 Human Bladder Carcinoma Cells (섬수 추출물에 의한 T24 인체 방광암세포의 증식억제에 관한 연구)

  • Park Tae Yeol;Park Cheol;Yoon Hwa Jung;Choi Yung Hyun;Ko Woo Shin
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.5
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    • pp.1449-1455
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    • 2004
  • Bufonis venenum (dried toad venom; Chinese name, Chan su) is a traditional Chinese medicine obtained from the skin venom gland of the toad. It has long been used in treating arrhythmia and other heart diseases in China and other Asian countries. Additionally, Bufonis venenum has been reported to selectively inhibit the growth of various lines of human cancer cells. In the present study, it was examined the effects of extract of Bufonis venenum (EBV) on the growth of human bladder carcinoma cell line T24 in order to investigate the anti-proliferative mechanism and induction of apoptosis by EBV. Treatment of T24 cells to EBV resulted in the growth inhibition, morphological change and induction of apoptotic cell death in a dose-dependent manner. Flow cytometric analysis revealed that EBV treatment caused G2/M phase arrest of the cell cycle and down-regulation of cyclin A, cyclin B1 and Cdc2, which was associated with a marked up-regulation of cyclin-dependent kinases (Cdks) inhibitor p21 (WAF1/CIP1) in a p53-independent manner. The Cdc25C expression was also significantly inhibited by EBV treatment, however Wee1 kinase expression was not affected. The induction of apoptotic cell death by EBV was connected with down-regulation of anti-apoptotic Bcl-XS/L expression without alteration pro-apoptotic Bax expression. Taken together, these findings suggest that EBV may be a potential chemotherapeutic agent for the control of human bladder carcinorma cells and further studies will be needed to identify the active compounds that confer the anti-cancer activity of EBV.