• 제목/요약/키워드: Nuclear localization

검색결과 308건 처리시간 0.025초

Expression and Localization of Heat Shock Protein 70 in Frozen-thawed IVF and Nuclear Transferred Bovine Embryos

  • Chung, K.S.;Choi, Y.J.;Song, S.J.;Do, J.T.;Yoon, B.S.;Kim, Y.J.;Lee, H.T.
    • 한국가축번식학회지
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    • 제26권4호
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    • pp.311-320
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    • 2002
  • The objective of this study was to assess the developmental potential in vitro produced embryos frozen-thawed with the various containers, and also examined expression and localization of heat shock protein 70 at these embryos. For the vitrification, 2-cell, 8-cell and blastocyst stage embryos produced by in vitro fertilization (IVF) and nuclear transfer (NT) were exposed the ethylene glycol 5.5 M freezing solution (EC 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop, and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min. and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid and cryo-loop. However, survival rates by straw were relatively lower than other containers. The use of cryo-loop resulted in only survival of nuclear transferred embryos (43.7%). Also, there embryos after IVF or NT were analysed by semi-quantitive reverse transcription-polymerase chain reaction (RT- PCR) methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNh were higher thawed embryos than control embryos. Immunocytochemistry used to localize the hsp 70 protein in embryos. Two and 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some frozen-thawed embryos. However, in the control, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform In distribution. Therefore, this result suggests that the exploiting Hsp 70 in the early embryos may be role for protection of stress condition for increase viability of embryos within IVF, NT and there frozen-thawed embryos.

Regulation of Nek6 Functions by Its SUMOylation on the $K^{252}$ Residue

  • Lee, Eun-Jeoung;Hyun, Sung-Hee;Chun, Jae-Sun;Shin, Sung-Hwa;Lee, Kyung-Eun;Park, In-Suk;Kang, Sang-Sun
    • Animal cells and systems
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    • 제11권2호
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    • pp.205-213
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    • 2007
  • Nek6 belongs to NIMA1 (never in mitosis, gene A) related kinase, which was originally identified in Aspergillus nidulans as a serine/threonine kinase critical for cell cycle progression. We noticed that the putative SUMOylation site is localized on the $K^{252}$ residue in $^{251}FKsD^{254}$ of Nek6, based on the consensus sequence ${\Phi}KxE$; where ${\Phi}$ represents L, I, V or F and x is any amino acid. We observed that the Nek6 SUMO mutant (K252R) has decreased protein kinase activity, nuclear speckle localization and protein stability, compared with that of the Nek6 wild type. However, the Nek6 SUMO mutant increased the cell survival rate of COS-1 cells as determined by FACS analysis. Therefore, our data suggest that SUMOylation on the $K^{252}$ residue of Nek6 is required for its normal functions, such as proper nuclear localization, kinase activity and protein stability, to control cell cycle.

The Expression and the Subcellular Localization of Regulatory Subunits of Class IA Phosphoinositide 3-Kinase in L6 Skeletal Muscle Cell

  • Woo Joo-Hong;Lim Jeong-Soon;Kim Hye-Sun
    • 대한의생명과학회지
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    • 제12권3호
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    • pp.201-208
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    • 2006
  • PI3-kinase activity through p85, the regulatory subunit of class IA PI3-kinase, is indispensable for the growth, differentiation, and survival of skeletal muscle cells, but little is known about the function of other regulatory subunits such as p55 and p50. We examined the subcellular localization and the expression of the regulatory subunits of class IA PI3-kinase in L6 myoblasts. Both p55 and p50 as well as p85 were expressed in L6 myoblasts. Whereas p85 was localized at both cytosolic and nuclear tractions, p55 and p50 were localized at only the nuclear traction. During the differentiation of L6 myoblasts, the protein concentrations of both p55 and p50 were decreased but that of p85 was not significantly changed. Menadione-induced oxidative stress induced the translocation of p85 from cytosol to nucleus and the increase of p55 expression. These results suggest that the regulatory subunits of class IA PI3-kinase play an important role in L6 myoblasts.

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Increased Expression of Epithelial Cell Adhesion Molecule (EpCAM) in Rat Hepatic Tumors Induced by Diethylnitrosamine

  • Kang, Jin Seok
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권8호
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    • pp.3627-3630
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    • 2012
  • The epithelial cell adhesion molecule (EpCAM) is a pan-epithelial differentiation antigen that is expressed on almost all carcinomas. However, a role in rat liver carcinogenesis has never been reported previously. Thus, its expression was investigated herein in rat liver tumors induced by diethylnitrosamine (DEN). Twenty male 5-week-old F344 rats were used in this experiment. Mini-osmotic pumps containing doses of 47.5 mg of DEN were inserted into the abdominal cavity of each animal to initiate liver carcinogenesis. All animals were sacrificed at 26 weeks after DEN treatment. At necropsy, hepatic masses were processed for histopathological examination, which revealed forty-four hepatocellular adenomas (HCAs) and twenty hepatocellular carcinomas (HCC). Tumors were immunohistochemically analyzed for EpCAM, proliferating cell nuclear antigen (PCNA) and co-localization of the two. EpCAM expression was mainly detected in hepatic tumor cells, showing a cytoplasmic staining pattern. However, expression was also slightly observed in normally-appearing surrounding hepatic cells. PCNA expression was highly detected in tumor cells, showing nuclear staining. Double staining of EpCAM and PCNA in tumors showed many cells with co-localization. Taken together, EpCAM and PCNA expression were increased in DEN-induced tumors and many tumor cells showed co-expression. It is suggested that EpCAM may increase during DEN-induced tumors, possibly associated with cell proliferation.

Fraxetin Induces Heme Oxygenase-1 Expression by Activation of Akt/Nrf2 or AMP-activated Protein Kinase α/Nrf2 Pathway in HaCaT Cells

  • Kundu, Juthika;Chae, In Gyeong;Chun, Kyung-Soo
    • Journal of Cancer Prevention
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    • 제21권3호
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    • pp.135-143
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    • 2016
  • Background: Fraxetin (7,8-dihydroxy-6-methoxy coumarin), a coumarin derivative, has been reported to possess antioxidative, anti-inflammatory and neuroprotective effects. A number of recent observations suggest that the induction of heme oxygenase-1 (HO-1) inhibits inflammation and tumorigenesis. In the present study, we determined the effect of fraxetin on HO-1 expression in HaCaT human keratinocytes and investigated its underlying molecular mechanisms. Methods: Reverse transcriptase-PCR and Western blot analysis were performed to detect HO-1 mRNA and protein expression, respectively. Cell viability was measured by the MTS test. The induction of intracellular reactive oxygen species (ROS) by fraxetin was evaluated by 2′,7′-dichlorofluorescin diacetate staining. Results: Fraxetin upregulated mRNA and protein expression of HO-1. Incubation with fraxetin induced the localization of nuclear factor-erythroid-2-related factor-2 (Nrf2) in the nucleus and increased the antioxidant response element-reporter gene activity. Fraxetin also induced the phosphorylation of Akt and AMP-activated protein kinase $(AMPK){\alpha}$ and diminished the expression of phosphatase and tensin homolog, a negative regulator of Akt. Pharmacological inhibition of Akt and $AMPK{\alpha}$ abrogated fraxetin-induced expression of HO-1 and nuclear localization of Nrf2. Furthermore, fraxetin generated ROS in a concentration-dependent manner. Conclusions: Fraxetin induces HO-1 expression through activation of Akt/Nrf2 or $AMPK{\alpha}/Nrf2$ pathway in HaCaT cells.

Apoptin gene delivery by a PAMAM dendrimer modified with a nuclear localization signal peptide as a gene carrier for brain cancer therapy

  • Bae, Yoonhee;Lee, Jell;Kho, Changwon;Choi, Joon Sig;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
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    • 제25권5호
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    • pp.467-478
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    • 2021
  • In this study, we aimed to synthesize PAMAMG3 derivatives (PAMAMG3-KRRR and PAMAMG3-HKRRR), using KRRR peptides as a nuclear localization signal and introduced histidine residues into the KRRR-grafted PAMAMG3 for delivering a therapeutic, carcinoma cell-selective apoptosis gene, apoptin into human primary glioma (GBL-14) cells and human dermal fibroblasts. We examined their cytotoxicity and gene expression using luciferase activity and enhanced green fluorescent protein PAMAMG3 derivatives in both cell lines. We treated cells with PAMAMG3 derivative/apoptin complexes and investigated their intracellular distribution using confocal microscopy. The PAMAMG3-KRRR and PAMAMG3-HKRRR dendrimers were found to escape from endolysosomes into the cytosol. The JC-1 assay, glutathione levels, and Annexin V staining results showed that apoptin triggered cell death in GBL-14 cells. Overall, these findings indicated that the PAMAMG3-HKRRR/apoptin complex is a potential candidate for an effective nonviral gene delivery system for brain tumor therapy in vitro.

Reactor Condition Monitoring via Wavelet Transform De-noising

  • Park, Chang-Je;Cho, Nam-Zin
    • 한국원자력학회:학술대회논문집
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    • 한국원자력학회 1996년도 추계학술발표회논문집(1)
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    • pp.67-72
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    • 1996
  • Wavelets are localized in space and in frequency. This localization properties result from the multiresolution analysis of wavelets. The wavelet transform can be used to detect singularity of dynamic systems after the signal is de-noised. We applied the wavelet transform decomposition and do-noising procedures to the Hanaro dynamics consisting of 39 nonlinear differential equation plus Gaussian noise. The numerical tests demonstrate that the wavelet transform de-noising is effective for detection of the abrupt reactivity change and computationally efficient. Thus this wavelet theory could be profitably utilized in a real-time system for automatic event recognition (e.g., reactor condition monitoring).

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원자력 등급용 안전방출밸브 개발 (The Development of Safety Relief Valve for Nuclear Service.)

  • 김칠성;김강태;김지헌;장기종;홍기성
    • 대한기계학회:학술대회논문집
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    • 대한기계학회 2003년도 춘계학술대회
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    • pp.629-636
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    • 2003
  • The purpose of this study is localization of safety relief valves for Nuclear Service through technical development with overall design, fabrication, inspection, capacity certification test and functional qualification test of safety relief valves in accordance with ASME Section III and KEPIC Code. Safety relief valve is the important equipment used to protect the pressure vessel, the steam generator and the other pressure facility from overpressure by discharging the operating medium when the pressure of system is reaching the design pressure of the system. But we're depending on technology of the other country up to the present time. Because we don‘ have our own technologies, we have been spent the great time and money on installing and repairing safety relief valve at nuclear power plant. Therefore we have to achieve the development of safety relief valves for Nuclear Service with our own technologies.

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Tc-99m HMPAO WBC 스캔에서 섭취 증가 소견을 보인 연골 육종 1례 (Increased Uptake of Tc-99m HMPAO WBC in Chondrosarcoma)

  • 강현구;최윤영;조석신
    • 대한핵의학회지
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    • 제35권6호
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    • pp.389-392
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    • 2001
  • Tc-99m HMPAO WBC scintigraphy is a highly sensitive and specific method for detection and localization of infection and abscess formation. Nevertheless, WBC accumulation in some neoplastic lesions has been reported. We experienced a case with unusual increased uptake of Tc-99m HMPAO WBC in chondrosarcoma. Although the exact cause ms unclear, the possible mechanism is discussed.

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발전소 설계 인력 수요의 통계적 분석 (Manpower Simulation for the Power Plant Design Engineering)

  • Byung Soo Moon;Poong Eil Juhn
    • Nuclear Engineering and Technology
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    • 제14권1호
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    • pp.1-9
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    • 1982
  • 이 논문에서는 Sargent & Lundy 회사로부터 입수한 발전소 설계 인력 분포에 관한 49개 실적치 커브와 Bechtel사가 제안한 수 개의 모델 커브를 분석하였다. 이들 커브로 부티 발전소 설계에 소요되는 인력분포를 Gamma분포로 가정하고 그 확률함수를 하나의 모델커브로 선정하였다. 이 모델커브를 이용하여 정부의 장기 원자력 발전소 설계계획에 따른 인력 수요를 년도별로 산출하였다. 또한 감응도 분석의 하나로서 엔지니어들의 숙련도에 따른 설계 인력 수요의 변화를 분석하였다.

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