• Title/Summary/Keyword: spindle assembly

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Cellular Toxic Effects and Action Mechanisms Of 2,2', 4,6,6'-Pentachlorobiphenyl

  • Kim Sun-Hee;Shin Kum-Joo;Kim Dohan;Kim Yun-Hee;Ryu Sung Ho;Suh Pann-Ghill
    • 한국생물공학회:학술대회논문집
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    • 2004.07a
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    • pp.1-20
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    • 2004
  • Polychlorinated biphenyls (PCBs), one a group of persistent and widespread environmental pollutants, have been considered to be involved in immunotoxicity, carcinogenesis, and apoptosis. However, the toxic effects and physical properties of a PCB congener are dependent on the structure. In the present study, we investigate the toxic effects and action mechanisms of PCBs In cells. Among the various congeners tested, 2,2',4,6,6'-PeCB-pentachlorobiphenyl (PeCB), a highly ortho-substituted congener having negligible binding affinity for aryl hydrocarbon receptor (AhR), caused the most potent toxicity and specific effects in several cell types. 2,2',4,6,6'-PeCB induced apoptotic cell death of human monocytic cells, suggesting that PCB-induced apoptosis may be linked to immunotoxicity. In addition, 2,2',4,6,6'-PeCB induced mitotic arrest by interfering with mitotic spindle assembly in NIH3T3 fibroblasts, followed by genetic instability which triggers p53 activation. Which suggests that 2,2',4,6,6'-PeCB may be involved in cancer development by causing genetic instability through mitotic spindle damage. On the other hand, 2,2',4,6,6'-PeCB increased cyclooxygenase-2 (COX-2) involved in cell survival through ERK1/2 MAPK and p53 in Rat-1 fibroblasts and mouse embryonic fibroblasts, triggering compensatory mechanism for abating its toxicity. Taken together, these results demonstrate that PCB congeners of different structure have distinct mechanism of action and 2,2',4,6,6'-PeCB causes several toxicity as well as compensatory mechanism in cells.

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IMMUNOHISTOCHEMICAL STUDY OF AURORA-2 KINASE IN THE ORAL SQUAMOUS CELL CARCINOMA (구강편평상피암종에서 Aurora-2 kinase 발현에 대한 면역조직화학적 연구)

  • Han, Se-Jin;Kim, Se-Woong;Kim, Kyung-Wook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.32 no.2
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    • pp.112-117
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    • 2010
  • Aurora kinases represent a novel family of serine/threonine kinases crucial for cell cycle control. Aurora-2 kinase is mainly involved in centrosome function, mitotic entry, and spindle assembly. Aurora-2 kinase overexpression causes centrosome amplification and the formation of multipolar mitotic spindles, which leads to tumor aneuploidy and so it has been found to play an important role in tumorigenicity in many cancers such as colorectal cancer, breast cancer and cervical cancer. Hence, the goal of this study is to identify the correlation of clinicopathlogical factors and overexpression of Aurora-2 kinase in oral squamous cell carcinoma. We studied the immunohistochemical staining of Aurora-2 kinase in 20 specimens of 20 patients with oral squamous cell carcinoma and the relationships between Aurora-2 kinase over expression and each of the clinico-pathological parameters were analyzed by Pearson correlation analysis. Statistical significance was set at P < 0.05. The results were as follows. 1. In the immunohistochemical study of poorly differentiated and invasive oral squamous cell carcinoma, the high level staining of Aurora-2 kinase was observed. 2. The correlation between immunohistochemical Aurora-2 kinase expression and histopathological differentiation of specimens was significant. These findings suggest that overexpression of Aurora-2 kinase may play a important role in carcinogenesis of oral squamous cell carcinoma.

Functional Analyses of Centrosomal Proteins, Nek2 and NuMA in Development of Mouse Gametes and Early Embryos

  • Youn, Hong-Hee;Oh, Hwa-Soon;Lee, Kwang-Hee;Son, Chae-Ick;Lee, Sang-Ho
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.96-96
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    • 2003
  • Nek2 (NIMA-related protein) is a mammalian cell cycle-regulated kinase that involves in chromosome condensation and centrosome regulation and NuMA (nuclear mitotic apparatus protein) is involved in spindle assembly during a cell cycle. The cellular distribution and organization of the centrosomal components is completely unknown during fertilization and embryonic development. We examined distribution of two well-known centrosomal proteins, Nek2 and NuMA in mouse gametes and embryos to get an insight in the reorganization of centrosomal proteins during germ cell development and early fertilization. Spermatogenic cells, gametes, and embryos were analyzed with anti-Nek2 or -NuMA antibodies by immunological assay, RT-PCR, and overexpression through gene transfection. Mitotically or meiotically active spermatogenic cells were intensively stained with these antibodies in both centrosomes and cytoplasm, whereas the oocytes showed different staining patterns depending on the meiotic stages. During maturation, GV, GVBD, and MI stage were clearly stained with NuMA antibody in the nucleus or cytoplasm at MII. Also, Nek2 was detectable in cytoplasm as scattered spots or chromosome associated at MII. In early developmental embryo, NuMA was detected in nucleus of each blastomere, while Nek2 was detected in cytoplasm. In contrast to previously reported results, Nek2 and NuMA were detected in both decondensing head, and the centriole of demembranated and decondensed sperm or whole body of trypsin-treated sperm for Nek2. During meiotic progress in oocytes, transcripts levels were the highest in MI stage and then downregulated in MII. Also, it shows dramatically change in early developmental embryos, firstly, it was increased until 4 cell stage and reduced in 8 cell stage, and finally, transcript levels were upregulated until blastoscyst. This finding suggests that cnetrosomal component may play an important role in reorganizing of functional centrosome during fertilization process and subsequent development.

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SEPT12 Interacts with SEPT6 and This Interaction Alters the Filament Structure of SEPT6 in Hela Cells

  • Ding, Xiangming;Yu, Wenbo;Liu, Ming;Shen, Suqin;Chen, Fang;Wan, Bo;Yu, Long
    • BMB Reports
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    • v.40 no.6
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    • pp.973-978
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    • 2007
  • Septins are a family of conserved cytoskeletal GTPase forming heteropolymeric filamentous structure in interphase cells, however, the mechanism of assembly are largely unknown. Here we described the characterization of SEPT12, sharing closest homology to SEPT3 and SEPT9. It was revealed that subcelluar localization of SEPT12 varied at interphase and mitotic phase. While SEPT12 formed filamentous structures at interphase, it was localized to the central spindle and to midbody during anaphase and cytokinesis, respectively. In addition, we found that SEPT12 can interact with SEPT6 in vitro and in vivo, and this interaction was independent of the coiled coil domain of SEPT6. Further, co-expression of SEPT12 altered the filamentous structure of SEPT6 in Hela cells. Therefore, our result showed that the interaction between different septins may affect the septin filament structure.

A Study on the Development and Surface Roughness of Roller Cam SCM415 by 5-Axis Machining (5축 가공에 의한 SCM415 롤러 캠 개발과 표면조도 연구)

  • Kim, Jin Su;Lee, Dong Seop;Kang, Seong Ki
    • Journal of the Korean Society for Precision Engineering
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    • v.30 no.4
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    • pp.397-402
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    • 2013
  • In this study, we carried out the each lines of section, using GC (green silicon carbide) whetstone, the SCM415 material which separated by after and before heat treatments process, in 3+2 axis machining centers for integrated grinding after cutting end mill works, the spindle speed 8000 rpm and feed rate 150 mm/min. For the analysis of the centerline average roughness (Ra), we measured by 10 steps stages. Using Finite element analysis, we found the result of the load analysis effect of the assembly parts, when applied the 11 kg's load on both side of the ATC (Automatic tool change) arm. The result is as follows. For the centerline average roughness (Ra) in the non-heat treatment work pieces, are appeared the most favorable in the tenth section are $0.510{\mu}m$, that were shown in the near the straight line section which is the smallest deformation of curve. In addition, the bad surface roughness appears on the path is to long by changing angle, the more inclined depth of cut, because the chip discharging is not smoothly.

Caffeine Treatment during Oocyte Aging Improves the Developmental Rate and Quality in Bovine Embryos Developing In Vitro

  • Choi, Hyun-Yong;Lee, Sung-Hyun;Xu, Yong-Nan;Lee, Seung-Eun;Kim, Nam-Hyung
    • Reproductive and Developmental Biology
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    • v.37 no.4
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    • pp.281-287
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    • 2013
  • In mammal, unfertilized oocytes remain in the oviduct or under in vitro culture, which is called "oocyte aging". This asynchrony negatively affects fertilization in pre- and post-implantation embryo development. Caffeine a phosphodiesterase inhibitor is known to rescue oocyte aging in several species. The objective of this study is to determine the cytoskeleton distribution in aged oocytes and the embryo developmental ability of aged oocytes in the present or absence of caffeine during maturation. Caffeine treatment increased the incidence of normal spindle assembly of aged oocytes (treatment, $67.57{\pm}4.11%$ aging, $44.61{\pm}6.4%$) and no significant differences compared to control group. Fluorescence values were compared using ROS (Reactive oxidation species) stain. Fluorescence values appear of control group intensity rate ($51.53.{\pm}3.80$), aging group ($68.10{\pm}5.54$) and treatment of caffeine ($45.04{\pm}2.98$). Aged oocytes that were derived from addition of caffeine to the IVM (in vitro maturation) medium had significantly increased 2-cell that developed to the blastocyst stage compared to the aging group. Blastocysts, derived from caffeine treatment group, significantly increased the total cell number compare aging ($90.44{\pm}10.18$ VS $67.88{\pm}7.72$). Apoptotic fragments of genomic DNA were measured in individual embryo using TUNEL assay. Blastocyst derived from caffeine treatment group decreased significantly the apoptotic index compared to blastocyst derived from aging group. In conclusion, we inferred that the caffeine treatment during oocyte aging can improve the developmental rate and quality in bovine embryos developing in vitro.

The Study of Spontaneous Developmental Abnormalities and Toxicology of Benomyl and Its Metabolite on Salamander, Hynobius leechii.

  • Park, Yong-Uk
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2005.12a
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    • pp.38-45
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    • 2005
  • The egg bags of Korean salamander(Hynobius leechii) were collected from farmlands in Gyeongsangnam-do area. The assumed breeding time, numerical variation of embryos in each egg bag, mortality and the rates of abnormalities were investigated. The toxicity of benomyl, the metabolite carbendazim and BIC which were frequently spread in agricultural area and caused spontaneous embryonic malformation was investigated. The assumed breeding time between the end of February and the end of March has the difference about a month because of a habitat and it takes about 2 or 3 weeks from laying eggs to hatching. The length of each egg bag and the number of embryos were very varied in each area. It is due to geographical variation. Among egg bags in total study area, only 406 of egg bags(17.70% of total egg bags) developed all of embryos to normal larvae, and 78.49% of total embryos were normally developed. The patterns of spontaneous embryonic malformation were 26 species from A to Z and the abnormal patterns in individual were 8 species and above. the geographical differences about the abnormal pattern were identified and 11 habitats categorized 4 groups. The most frequent abnormality in Gyeongsangnam-do area is the dysplasia of external gill. The caudal dysplasia, abdominal blister and dysplasia of fin were also frequently observed. Individuals showing severe external defect were histologically studied and they showed retinal hypo-pigmentation, thyroid carcinoma, somatic muscular dysplasia, degeneration of cephalic neuron and various organ dysplasia. Benomyl and carbendazim were treated by 10pM$^{\sim}$10uM and BIC was treated by 1$^{\sim}$40ppm to know the effect of toxicity about toxic substance of salamander. After benomyl was treated, a survival rate was sharply dropped from 2 to 8 days. $LC_{100}$ identified in $1{\mu}M$, $LC_{50}$ identified between 100nM and $1{\mu}M$. $EC_{50}$ was assumed between 10nM and 100nM. The prevalent external malformation was abdomen swelled abnormally and histo-pathological effects were abdomen, neural tube and lens hernia. This suggests that benomyl is the toxicitic substance which inhibits the development of digestive system and nervous system. The result of treated carbendazim was similar to that of the treated benomyl. The survival rate is sharply dropped between 2 and 6 days. $LC_{100}$ was identified $1{\mu}M$ and $LC_{50}$ was identified between 10nM and 100nM. This shows that cabendazim has stronger lethal toxicity than benomyl. Ventral blister, eye dysplasia and cephalic dysplasia in the individual of external malformation mean that cabendazim affected nervous system much more than benomyl. Because the toxicity of BIC affected less in the beginning but affected more in the near hatching period, the period causing toxicity is somewhat different. $LC_{100}$ identified near 40ppm and $LC_{50}$ identified near 25ppm. The external defect shows mainly ventral blister and histo-pathological results show intestinal deformities. This result suggests the BIC inhibited strongly the development of digestive system. These abnormal developments may be caused by antimitotic action, inhibition of tubulin complex, destruction of microtubule, inhibitions of neurulation and closing of neural fold, and by the inhibition of movement of neural crest cells of benomyl. These abnormal developments may be caused by the rupture of epithelium, the loss of microtubule, the reduction of spindle size, the inhibition of spindle assembly formation, the destruction of spindle poles of carbendazim. These abnormal developments may be caused cytotoxicity by inhibition of the synthesis of a number of macromolecules and similar reaction the inhibition of benomyl.

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Finite Element Analysis on the Deformation Behavior Safety of a Gas Valve (가스밸브의 변형거동 안전성에 관한 유한요소해석)

  • Kim, Chung Kyun
    • Journal of the Korean Institute of Gas
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    • v.21 no.4
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    • pp.70-75
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    • 2017
  • This paper presents the FEM analysis results on the deformation behavior safety of automatic cut-off horizontal and conventional vertical gas valves. Based on the FEM analysis, the primary maximum deformation of $4.4{\mu}m$ was formed on the right end side of a valve body when the internal gas pressure was supplied on the screw port and gas discharge port of an automatic cut-off horizontal gas valve. And the secondary maximum deformation of $2.9{\mu}m$ was formed on the end side of safety valve port. This small deformation of an automatic cut-off horizontal gas valve is strongly related to the balanced design of a horizontal gas valve main body, which is composed of a screw part, gas outlet port, port for a stem and spindle shaft assembly, and safety valve port. But, the primary maximum deformation of 0.076mm was formed on the upper part of a conventional automatic cut-off vertical gas valve when the internal gas pressure was supplied on the screw port and gas discharge port. And the secondary maximum deformation of 0.055mm was formed on the left end side of a gas outlet port. This may effect on the sealing clearance of o-ring that is inserted on the groove of an automatic cut-off unit. Thus, this paper recommends an automatic cut-off horizontal gas valve compared with that of a conventional gas valve for a gas leakage free mechanism of a LPG cylinder valve.