• Title/Summary/Keyword: GRP

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Neuroprotective Effect of Hwangryunhaedok-tang Against Glucose-deprivation Induced Cytotoxicity in PC-12 Cells (Glucose-deprivation으로 유발된 PC-12 cell의 세포 고사에 대한 황연해독탕(黃連解毒湯)의 보호 효과)

  • Kim, Hye-Yoon;Shin, Sun-Ho;Lee, In
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.6
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    • pp.1462-1469
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    • 2008
  • The purpose of the study was to confirm what effect HRHDT treatment had on cell extinction by damage of endoplasmic reticulum induced to PC-12 cell damage by glucose deprivation. The study confirmed what effect it had on forming the condition of glucose deprivation within a culture fluid of PC-12 cell and on a nerve cell's survival rates and tested whether HRHDT could prevent extinction of PC-12 cell by glucose deprivation. Also, the study confirmed what effect HRHDT treatment had on the emitted quantity of LDH by glucose deprivation. To examine PC-12 cell's behavioral change under the condition of glucose deprivation and a protective effect of HRHDT on the change, the study observed PC-12 cell's behavioral change with a microscope. Also, the study confirmed density of calcium ion within cells followed by a culture time in the condition of glucose deprivation with FACS and confirmed what effect HRHDT treatment had on the above density of calcium ion within cells. Finally, the study carried out the western blot and confirmed what effect HRHDT treatment had on revelation of GRP 78 and CHOP protein and a segmental type of aspase 12. In this study, HRHDT rescued PC-12 cells from glucose deprivation-induced cell death. HRHDT also prevents the LDH release, Ca++ accumulation, and morphological change, which was associated with the ER stress. Furthermore, HRHDT reduced the expression of ER chaperone (Grp78 and CHOP) proteins by glucose deprivation in PC-12 cells. These results suggest that HRHDT might provide a useful therapeutic strategy in treatment of the neurodegenerative diseases caused by glucose deprivation injuries.

An immunohistochemical study of the gastro-entero endocrine cells in the snakehead, Channa(ophicephalus) argus (가물치 위장관에 있어서 내분비세포의 면역조직화학적 연구)

  • Lee, Hyeung-sik;Lee, Jae-hyun
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.297-303
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    • 1996
  • The regional distribution and relative frequency of endocrine cells in the GIT of the snakehead, Channa(Oph icephalus) argus were studied immunohistochemically. Five kinds of endocrine cells and one kind of nerve cell were identified in this study. A few numbers of secretin-immunoreactive cells were restricted to the cardia and fundus of the stomach. Moti-lin- and GRP-immunoreactive cells were rare in the epithelium of the distal and proximal intestines. PYY-immunoreactive cells were found in a few number in the pyloric caeca and were rare in the distal intestine. Substance P-immunoreactive cells were distributed relatively numerous from the fundus to the distal intetsine. Also, their nerve cells were detected in occurrence in the lamina propria in the distal intestine. No neurotensin-, met-Enk- and GIP-immunoreactive cells were found in the GIT of the snakehead.

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Hull Strength Evaluation of Dissemination 12ft Bass Fishing Boat Using FEA (보급형 12피트 배스 낚시보트의 유한요소해석을 통한 선체강도평가)

  • Oh, Young-Cheol;Ko, Jae-Yong;Chung, Se-Yun;Choi, Jeong-Hwan;Kim, Hyeon-Jin
    • Proceedings of the Korean Institute of Navigation and Port Research Conference
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    • 2012.06a
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    • pp.225-226
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    • 2012
  • Recently, In the domestic is used mainly fiberglass reinforced plastic(GRP) and is built for maritime leisure vessel & structures. but it less attention is insufficient for ship of structural strength assessment of maritime leisure vessel. Therefore, The structural strength evaluation suitable for the domestic situation formulate and is applied to domestic regulations of "Guidance for Recreational Crafts" of Korea register of shipping(KR) & "Reinforced plastic line structure standards" of Ministry of Land, Transport and Maritime Affairs. the structural design and finite element analysis(FEA) to ensure the reliability.

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Bioproduction of trans-10,cis-12-Conjugated Linoleic Acid by a Highly Soluble and Conveniently Extracted Linoleic Acid Isomerase and an Extracellularly Expressed Lipase from Recombinant Escherichia coli Strains

  • Huang, Mengnan;Lu, Xinyao;Zong, Hong;Zhuge, Bin;Shen, Wei
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.739-747
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    • 2018
  • The low solubility and high-cost recovery of Propionibacterium acnes polyunsaturated fatty acid isomerase (PAI) are key problems in the bioproduction of high value-added conjugated linoleic acid (CLA). To improve the solubility of recombinant PAI, six chaperone proteins were coexpressed with PAI. Introduction of GroELS proteins dramatically improved the PAI solubility from 29% to 97%, with increased activity by 57.8%. Combined expression of DnaKJ-GrpE and GroELS proteins increased the activity by 11.9%. In contrast, coexpression of DnaKJ-GrpE proteins significantly reduced the activity by 57.4%. Plasmids pTf16 harboring the tig gene and pG-Tf2 containing the tig and groEL-groES genes had no visible impact on PAI expression. The lytic protein E was then introduced into the recombinant Escherichia coli to develop a cell autolysis system. A 35% activity of total intracellular PAI was released from the cytoplasm by suspending the lysed cells in distilled water. The PAI recovery was further improved to 81% by optimizing the release conditions. The lipase from Rhizopus oryzae was also expressed in E. coli, with an extracellular activity of 110.9 U/ml. By using the free PAI and lipase as catalysts, a joint system was established for producing CLA from sunflower oil. Under the optimized conditions, the maximum titer of t-10,c-12-CLA reached 9.4 g/l. This work provides an effective and low-cost strategy to improve the solubility and recovery of the recombinant intracellular PAI for further large-scale production of CLA.

A Study on Gamma ray effects on Stress Response and Cellular Toxicity using Bacterial Cells

  • Min, Ji-Ho;Lee, Hyeon-Ju;Lee, Chang-U;Gu, Man-Bok
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.187-190
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    • 2000
  • Effects of gamma ionizing radiation on recombinant Escherichia coli cells containing stress promoters, recA, fabA, grpE, or katG, fused to luxCDABE originated from Vibrio fischeri were characterized by monitoring transcriptional responses reflected by bioluminescent output. Quantification of gamma-ray intensity may be possible using the recA and fabA promoter fusion since a linear enhancement of bioluminescence emission with increasing gamma-ray intensity was observed. Other strains sensitive to either oxidative stress (DPD2511, katG::luxCDABE) or protein-damaging stress (TV1061, grpE::luxCDABE) were also irradiated by gamma-rays, and resulted in no noticeable bioluminescent output while DPD2794 with recA promoter and DPD2540 with fabA promoter irradiated by the same intensities of gamma-rays gave a significant bioluminescent output. This indicates that the main stresses in the recombinant bacteria caused by ionizing radiation are DNA and membrane-damage, not protein- or oxidative-damage. In addition, in this study, to investigate the relationship between the radiation dose rate and bacterial responses, two recombinant Escherichia coli strains, DPD2794 and GC2, containing lac promoter fused to luxCDABE originating from Photorhapdus luminescences, were used for detecting DNA damage and cellular toxicity under various radiation dose rates. Throughout this study, it was found that these bacteria showed quantitative stress responses to DNA damage and general toxicity caused by gamma rays, depending on the radiation dose rates, indicating that the bacterial stress responses and general toxicity were seriously influenced according to radiation dose rates.

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Effects of Parafibromin Expression on the Phenotypes and Relevant Mechanisms in the DLD-1 Colon Carcinoma Cell Line

  • Zhao, Shuang;Sun, Hong-Zhi;Zhu, Shi-Tu;Lu, Hang;Niu, Zhe-Feng;Guo, Wen-Feng;Takano, Yasuo;Zheng, Hua-Chuan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4249-4254
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    • 2013
  • Background: Parafibromin is a protein encoded by the HRPT2 (hyperparathyroidism 2) oncosuppressor gene and its down-regulated expression is involved in pathogenesis of parathyroid, breast, gastric and colorectal carcinomas. This study aimed to clarify the effects of parafibromin expression on the phenotypes and relevant mechanisms of DLD-1 colon carcinoma cells. Methods: DLD-1 cells transfected with a parafibromin-expressing plasmid were subjected to examination of phenotype, including proliferation, differentiation, apoptosis, migration and invasion. Phenotype-related proteins were measured by Western blot. Parafibromin and ki-67 expression was detected by immunohistochemistry on tissue microarrays. Results: The transfectants showed higher proliferation by CCK-8, better differentiation by electron microscopy and ALP activity and more apoptotic resistance to cisplatin by DNA fragmentation than controls. There was no difference in early apoptosis by annexin V, capase-3 activity, migration and invasion between DLD-1 cells and their transfectants. Ectopic parafibromin expression resulted in down-regulated expression of smad4, MEKK, GRP94, GRP78, $GSK3{\beta}$-ser9, and Caspase-9. However, no difference was detectable in caspase-12 and -8 expression. A positive relationship was noted between parafibromin and ki-67 expression in colorectal carcinoma. Conclusions: Parafibromin overexpression could promote cell proliferation, apoptotic resistance, and differentiation of DLD-1 cells.

Four Members of Heat Shock Protein 70 Family in Korean Rose Bitterling (Rhodeus uyekii)

  • Kim, Jung Hyun;Dong, Chun Mae;Kim, Julan;An, Cheul Min;Baek, Hae Ja;Kong, Hee Jeong
    • Development and Reproduction
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    • v.19 no.3
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    • pp.135-144
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    • 2015
  • Heat shock protein (HSP) 70, the highly conserved stress protein families, plays important roles in protecting cells against heat and other stresses in most animal species. In the present study, we identified and characterized four Hsp70 (RuHSP4, RuHSC70, RuHSP12A, RuGRP78) family proteins based on the expressed sequence tag (EST) analysis of the Korean rose bitterling R. uyekii cDNA library. The deduced RuHSP70 family has high amino acid identities of 72-99% with those of other species. Phylogenetic analysis revealed that RuHsp70 family clustered with fish groups (HSP4, HSC70, HSP12A, GRP78) proteins. Quantitative RT-PCR analysis showed the specific expression patterns of RuHsp70 family members in the early developmental stages and several tissues in Korean rose bitterling. The expression of 4 groups of Hsp70 family was detected in all tested tissue. Particularly, Hsp70 family of Korean rose bitterling is highly expressed in hepatopancreas and sexual gonad (testis and ovary). The expression of Hsp70 family was differentially regulated in accordance with early development stage of Rhodeus uyekii.

ER Stress-Induced Jpk Expression and the Concomitant Cell Death

  • Kim Hye Sun;Chung Hyunjoo;Kong Kyoung-Ah;Park Sungdo;Kim Myoung Hee
    • Biomedical Science Letters
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    • v.11 no.2
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    • pp.135-141
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    • 2005
  • A Jopock (Jpk), a trans-acting factor associating with the position-specific regulatory element of murine Hoxa-7, has shown to have a toxicity to both prokaryotic and eukaryotic cells when overexpressed. Since Jpk protein harbors a transmembrane domain and a putative endoplasmic reticulum (ER)-retention signal at the N-terminus, a subcellular localization of the protein was analyzed after fusing it into the green fluorescent protein (GFP): Both N-term (Jpk-EGFP) and C-term tagged-Jpk (EGFP-Jpk) showed to be localized in the ER when analyzed under the fluorescence microscopy after staining the cells with ER- and MitoTracker. Since ER stress triggers the ER-stress mediated apoptosis to eliminate the damaged cells, we analyzed the expression pattern of Jpk under ER-stress condition. When MCF7 cells were treated with the ER-stress inducer such as DTT and EGTA, the expression of Jpk was upregulated at the transcriptional level like that of Grp78, a molecular chaperone well known to be overexpressed under ER-stress condition. In the presence of high concentration of ER-sterss inducer (10 mM), about 70 (DTT) to $95\%$ (EGTA) of cells died stronly expressing ($10\~12$ fold) Jpk. Whereas at the low concentration ($0.001\~1.0\;mM$) of the inducer, the expression of Jpk was increased about 2.5 (EGTA) to 5 fold (DTT), which is rather similar to those of ER chaperone protein Grp78. These results altogether indicate that the ER-stress upregulated the expression of Jpk and the excess stress induces the ER-stress induced apoptosis and the concomitant expression of Jpk.

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Changes in the Contents of Prosapogenin in Ginseng Radix Palva (Panax ginseng) Depending on the Extracting Conditions (미삼의 추출 조건에 따른 인삼 프로사포게닌 성분 변화)

  • Lee, Sun-A;Jo, Hee-Kyung;Sung, Min-Chang;Cho, Soon-Hyun;Song, You-Chan;Im, Byung-Ok;Ko, Sung-Kwon
    • Korean Journal of Pharmacognosy
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    • v.43 no.2
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    • pp.152-156
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    • 2012
  • This study compared the contents of ginseng prosapogenin depending on the extracting conditions of Ginseng Radix palva(Panax ginseng) to provide basic information for developing Ginseng Radix palva-based functional foods. Our findings show that the content of crude saponin peaked at 18 hours of extraction and when extracted twice at $100^{\circ}C$ (GRP-18). However, the content of total saponin reached its height at 6 hours of extraction at $100^{\circ}C$ (GRP-6) and when extracted twice. On the other hand, the content of ginsenoside $Rg_3$, $Rg_5$ and $Rk_1$ from Red and Black ginseng reached their heights at 18 hours of extraction, followed by 72 hours and 15 hours of extraction at $100^{\circ}C$. And at $100^{\circ}C$ the main prosapogenin of the content of Black ginseng ginsenoside $Rg_5$ and $Rk_1$ reached their heights at 18 hours of extraction, followed by 72 hours and 15 hours of extraction.