• Title/Summary/Keyword: ${\gamma}$-Tubulin

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Three-Dimensional Quantitative Structure Activity Relationship Studies on the Flavone Cytotoxicity and Binding to Tubulin

  • Kim, Ja-Hong;Sohn, Sung-Ho;Hong, Sun-Wan
    • Journal of Photoscience
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    • v.8 no.3_4
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    • pp.119-121
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    • 2001
  • Three-Dimensional Quantitative Structure-Activity Relationship(QSAR) has been investigated over 67 flavonoids to correlate and predict GI$\sub$50/ values. The partial least-squares(PLS) model was performed to calculate the activity of each derivatives, and this was compared with the actual value. The results of the cross-validated(${\gamma}$$^2$=0.997) values show that cytotoxic activities play an important role which is in good agreement with the observed GI$\sub$50/ values.

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Regulation of Apoptosis and Cell Cycle in Irradiated Mouse Brain (마우스의 대뇌조직에서 방사선에 의한 아포토시스와 세포주기의 조절)

  • Oh, Won-Yong;Song, Mi-Hee;Chung, Eun-Ji;Seong, Jin-Sil;Suh, Chang-Ok
    • Radiation Oncology Journal
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    • v.19 no.2
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    • pp.146-152
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    • 2001
  • Purpose : To investigate the regulation of apoptosis and cell cycle in mouse brain irradiation. Materials and Methods : 8-week old male mice, C57B1/6J were given whole body $\gamma-radiation$ with a single dose of 25 Gy using Cobalt 60 irradiator. At different times 1, 2, 4, 8 and 24hr after irradiation, mice were killed and brain tissues were collected. Apoptotic cells were scored by TUNEL assay. Expression of p53, Bcl-2, and Bax and cell cycle regulating molecules; cyclins Bl, Dl, E and cdk2, cdk4, $p34^{cdc2}$ were analysed by Western blotting. Cell cycle was analysed by Flow cytometry. Results : The peak of radiation induced apoptosis is shown at 8 hour after radiation. With a single 25 Gy irradiation, the peak of apoptotic index in C57B1/6J is $24.0{\pm}0.25$ (p<0.05) at 8 hour after radiation. Radiation upregulated the expression of p53/tubulin, Bax/tubulin, and Bcl-2/tubulin with 1.3, 1.1 and 1.45 fold increase, respectively were shown at the peak level at 8 hour after radiation. The levels of cell cycle regulating molecules after radiation are not changed significantly except cyclin D1 with 1.3 fold increase. Fractions of Go-Gl, G2-M and S phase in the cell cycle does not specific changes by time. Conclusion : In mouse brain tissue, radiation induced apoptosis is particularly shown in a specific area, subependyma. These results and lack of radiation induced changes in cell cycle ofter better understanding of radiation response of noraml brain tissue.

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Cloning and Functional Characterization of Ptpcd2 as a Novel Cell Cycle Related Protein Tyrosine Phosphatase that Regulates Mitotic Exit

  • Zineldeen, Doaa H.;Wagih, Ayman A.;Nakanishi, Makoto
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3669-3676
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    • 2013
  • Faithful transmission of genetic information depends on accurate chromosome segregation as cells exit from mitosis, and errors in chromosomal segregation are catastrophic and may lead to aneuploidy which is the hallmark of cancer. In eukaryotes, an elaborate molecular control system ensures proper orchestration of events at mitotic exit. Phosphorylation of specific tyrosyl residues is a major control mechanism for cellular proliferation and the activities of protein tyrosine kinases and phosphatases must be integrated. Although mitotic kinases are well characterized, phosphatases involved in mitosis remain largely elusive. Here we identify a novel variant of mouse protein tyrosine phosphatase containing domain 1 (Ptpcd1), that we named Ptpcd2. Ptpcd1 is a Cdc14 related centrosomal phosphatase. Our newly identified Ptpcd2 shared a significant homology to yeast Cdc14p (34.1%) and other Cdc14 family of phosphatases. By subcellular fractionation Ptpcd2 was found to be enriched in the cytoplasm and nuclear pellets with catalytic phosphatase activity. By means of immunofluorescence, Ptpcd2 was spatiotemporally regulated in a cell cycle dependent manner with cytoplasmic abundance during mitosis, followed by nuclear localization during interphase. Overexpression of Ptpcd2 induced mitotic exit with decreased levels of some mitotic markers. Moreover, Ptpcd2 failed to colocalize with the centrosomal marker ${\gamma}$-tubulin, suggesting it as a non-centrosomal protein. Taken together, Ptpcd2 phosphatase appears a non-centrosomal variant of Ptpcd1 with probable mitotic functions. The identification of this new phosphatase suggests the existence of an interacting phosphatase network that controls mammalian mitosis and provides new drug targets for anticancer modalities.

Nuclear and Microtubule Reorganization in Cattle Nuclear Transfered Embryos

  • Shin, Mi-Ra;Park, Sang-Wook;Cui, Xiang-Shun;Shim, Ho-Sup;Kim, Nam-Hyung
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.18-18
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    • 2001
  • Despite of importance of integrated events of nucleus and microtubule remodeling in nuclear transferred embryos with somatic cells, little information is available on this subject. In this study we configured chromatin and microtubule organization following somatic cell nuclear transfer in pre- and non-activated bovine oocytes in order to clearify nuclear remodeling process and to demonstrate centrosome inheritance during nuclear transfer. The cumulus-oocyte complexes were collected from slaughterhouse and were matured in vitro for 20 h in TCM 199 supplemented hormone. Matured bovine oocytes were enucleated by aspirating the frist polar body and metaphase chromatin using a beveled pipette. Bovine fibroblast cells were fused into enucleated oocyte by electrical stimulation. Reconstructed oocytes were activated with ionomycine and 6-dimethylaminopurin, and then cultured in CRlaa medium. The organization of nuclear and microtubules were observed using laser-scanning confocal microscopy. At 1 hour after fusion, microtubule aster was seen near the transferred nucleus in most oocytes regardless activation condition. While most of fibroblast nuclei remodeled to premature chromosome condensation (PCC) and to the two masses of chromosome in non-activated oocytes, a few number of fibloblasts went to PCC and multiple pronuclear like structures in activated oocytes. Microtubular spindle was seen around condensed chromosome. Gamma-tubulin was detected in the vicinity of condensed chromosome, suggesting this is a transient spindle. The spindle seperated nucleus into two masses of chromatin which developed to the pronuclear like structures. Two pronuclear like structures were than apposed by microtubular aster and formed one syngamy like nuclear structure at 15 h following nuclear transfer. At 17 to 18 h after fusion, two centrosomes were seen near the nucleus, which nucleates micrtubules for two cell cleavage. While 31% of reconstructed oocytes in non-activated condition developed to morulae and blastocysts, a few reconstructed oocytes in pre-activated condition developed to the blastocyst. These results suggested introduction of foreign centrosome during nuclear transfer, which appeared to give an important role for somatic cell nuclear reprogramming.

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Chk2 Regulates Cell Cycle Progression during Mouse Oocyte Maturation and Early Embryo Development

  • Dai, Xiao-Xin;Duan, Xing;Liu, Hong-Lin;Cui, Xiang-Shun;Kim, Nam-Hyung;Sun, Shao-Chen
    • Molecules and Cells
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    • v.37 no.2
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    • pp.126-132
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    • 2014
  • As a tumor suppressor homologue during mitosis, Chk2 is involved in replication checkpoints, DNA repair, and cell cycle arrest, although its functions during mouse oocyte meiosis and early embryo development remain uncertain. We investigated the functions of Chk2 during mouse oocyte maturation and early embryo development. Chk2 exhibited a dynamic localization pattern; Chk2 expression was restricted to germinal vesicles at the germinal vesicle (GV) stage, was associated with centromeres at pro-metaphase I (Pro-MI), and localized to spindle poles at metaphase I (MI). Disrupting Chk2 activity resulted in cell cycle progression defects. First, inhibitor-treated oocytes were arrested at the GV stage and failed to undergo germinal vesicle breakdown (GVBD); this could be rescued after Chk2 inhibition release. Second, Chk2 inhibition after oocyte GVBD caused MI arrest. Third, the first cleavage of early embryo development was disrupted by Chk2 inhibition. Additionally, in inhibitor-treated oocytes, checkpoint protein Bub3 expression was consistently localized at centromeres at the MI stage, which indicated that the spindle assembly checkpoint (SAC) was activated. Moreover, disrupting Chk2 activity in oocytes caused severe chromosome misalignments and spindle disruption. In inhibitor-treated oocytes, centrosome protein ${\gamma}$-tubulin and Polo-like kinase 1 (Plk1) were dissociated from spindle poles. These results indicated that Chk2 regulated cell cycle progression and spindle assembly during mouse oocyte maturation and early embryo development.

Effect of Pulsed Electromagnetic Field Stimulation on TNF-α/IFN-γ induced inflammatory response in human skin keratinocytes HaCaT Cell to reduce inflammatory factors (인간 피부각질세포 HaCaT Cell에서 TNF-α/IFN-γ로 유도된 염증 반응에 대한 펄스형 전자기장(PEMF) 자극의 염증 인자 완화 효과)

  • Jun Young Kim;Chan Ho Park;Chang Soon Park;Yong Heum Lee
    • Journal of Biomedical Engineering Research
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    • v.44 no.6
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    • pp.443-449
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    • 2023
  • The purpose of this study was to observe cell death in human keratinocytes stimulated against the infectious cytokines TNF-α and IFN-γ, and to observe the expression of Phospho-NF-κB due to phosphorylation of IkB to confirm the mechanism of inhibiting the expression of inflammatory cytokines. As a result of cell viability analysis, differences in PEMF stimulation time were observed little by little after 1 hour, 3 hours, 6 hours, 12 hours, 24 hours, and 48 hours, but there was no statistical significance according to PEMF stimulation time for each time (p>0.05). No significant difference was observed in the total amount of NF-κB present in the cytoplasm and nucleus, but a significant decrease in the expression of phosphorylated NF-κB was observed in the group exposed to PEMF stimulation for 24 hours (*p<0.05). The expression of IL-1β was observed in all inflammation-induced groups, and the concentration of IL-1β compared to α-Tubulin expression was reduced by about 54% in the PEMF-stimulated group for 24 hours compared to the control group (***p<0.001). As a result of the study, it is shown that PEMF stimulation does not negatively affect HaCaT cells from 0 to 48 hours and can inhibit the expression of inflammatory cytokines by inhibiting the pathway of NF-κB.

Distribution and Characteristics of Penicillium spp. in Meju, aKorean Traditional Fermented Soybean Brick (전통 메주에서의 Penicillium spp.의 분포 및 특징)

  • Kang Uk Kim;Jungho Lee;Shin Young Roh;Yong-Ho Choi;Byung-Serk Hurh;Inhyung Lee
    • Microbiology and Biotechnology Letters
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    • v.51 no.4
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    • pp.441-448
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    • 2023
  • Penicillium spp. are frequently found in meju, a Korean traditional fermented soybean brick. We isolated and identified 96 Penicillium spp. from 22 traditional meju, and their β-tubulin genes were sequenced for the genetic and taxonomic study. Penicillium Section Viridicata was the most commonly isolated group. Notably, we also isolated and identified Penicillium roqueforti, a crucial industrial strain employed in the fermentation of blue cheese. Additionally, certain strains exhibited relatively high protease and γ-glutamyl transpeptidase activities, suggesting that they might contribute to the development of kokumi flavor during meju fermentation. Interestingly, all eight Penicillium spp., including P. roqueforti, were found to possess both types of MAT1 genes. This intriguing finding suggests the feasibility of strain improvement through mating, thereby offering opportunities for industrial applications. Therefore, these studies pave the way for a deeper exploration of Penicillium's role in meju fermentation, potentially leading to the development of starters for producing plant-based cheese-flavored condiments.