• Title/Summary/Keyword: GRP7

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Construction of Candida antarctica Lipase B Expression System in E. coli Coexpressing Chaperones (대장균에서의 Chaperone 동시 발현을 통한 Candida antarctica Lipase B 발현 시스템 구축)

  • Jung, Sang-Min;Lim, Ae-Kyung;Park, Kyung-Moon
    • KSBB Journal
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    • v.23 no.5
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    • pp.403-407
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    • 2008
  • Recently, Candida antarctica lipase B (CalB) draws attention from industries for various applications for food, detergent, fine chemical, and biodiesel, because of its characteristics as an efficient biocatalyst. Since many industrial processes carry out in organic solvent and at high temperature, CalB, which is stable under harsh condition, is in demand from many industries. In order to reform CalB promptly, the expression system which has advantages of ease to use and low cost for gene libraries screening was developed using E. coli. The E. coli strains, Rosettagami with competence for enhanced disulfide bond formation, Novablue, and $DH5{\alpha}$, were exploited in this study. To obtain the soluble CalB, the pCold I vector expressing the cloned gene at $15^{\circ}C$ and the chaperone plasmids containing groES/groEL, groES/groEL/tig, tig, dnaK/dnaJ/grpE, and dnaK/dnaJ/grpE/groES/groEL were used for coexpression of CalB and chaperones. The colonies expressing functional lipase were selected by employing the halo plate containing 1% tributyrin, and the CalB expression was confirmed by SDS-PAGE. E. coli Rosettagami and $DH5{\alpha}$ harbouring groES/groEL chaperones were able to express soluble CalB effectively. From a facilitative point of view, E. coli $DH5{\alpha}$ is more suitable for further mutation study.

How Phenolic Composites were chosen - In Case of England (7) (페놀 컴포지트 실용화의 길 - 영국의 경우 (CASE STUDY 7))

  • Nomaguchi Kanemasa;Forsdyke Ken L.
    • Composites Research
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    • v.18 no.1
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    • pp.55-66
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    • 2005
  • Beside fire resistance, for Phenolic Composites(here-in-after PCs), in actual, moisture resistance are also needed when they are applied to components in wet uses such as in Naval vessels and in off-shore operations where direct water contact may be expected. PCs have so-called 'Micro-porous' when they are made by normal GRP manufacturing techniques, which causes much larger amounts of water absorption in them to higher level of around $7\%$. So, this figure, which indicate impending disaster in a polyester matrix laminate, has made some specifiers suspicious of the effect of the water on mechanical performance of PC in 'wet' service. To solve this problem, a large amount work has been carried out by workers in UK and US. This paper is an attempt to explain that the effect of the high water absorption of PCs is not the disaster that some 'prophets' may foretell. Let's open our next door fur better society much safetyfied by our own efforts with PCs. like in UK.

Variation of the structural stability for the sonar dome window in a naval vessel according to the state of the drain valve (소나돔 충, 배수 밸브의 상태에 따른 함정용 소나돔 윈도우의 구조안정도 변화)

  • Han, HyungSuk;Lee, KyungHyun;Park, SeongHo;Lim, YongSoo
    • Journal of Advanced Marine Engineering and Technology
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    • v.38 no.7
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    • pp.844-853
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    • 2014
  • Since the active sonar for a naval vessel is usually installed in a bulbous bow, GRP(Glass reinforced plastic) material with low density and high strength is used for the material of the sonar dome window in order to prohibit impact by slamming wave or foreign material in the sea. The structural safety of the sonar dome is varied according to the interior and exterior distributed pressure on the sonar dome. Therefore, the variation of the structural safety according to the pressure variation of the sonar dome window caused by the drain valve state is studied by CAE.

A Study on the Pipe Position Estimation in GPR Images Using Deep Learning Based Convolutional Neural Network (GPR 영상에서 딥러닝 기반 CNN을 이용한 배관 위치 추정 연구)

  • Chae, Jihun;Ko, Hyoung-yong;Lee, Byoung-gil;Kim, Namgi
    • Journal of Internet Computing and Services
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    • v.20 no.4
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    • pp.39-46
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    • 2019
  • In recently years, it has become important to detect underground objects of various marterials including metals, such as detecting the location of sink holes and pipe. For this reason, ground penetrating radar(GPR) technology is attracting attention in the field of underground detection. GPR irradiates the radar wave to find the position of the object buried underground and express the reflected wave from the object as image. However, it is not easy to interpret GPR images because the features reflected from various objects underground are similar to each other in GPR images. Therefore, in order to solve this problem, in this paper, to estimate the piping position in the GRP image according to the threshold value using the CNN (Convolutional Neural Network) model based on deep running, which is widely used in the field of image recognition, As a result of the experiment, it is proved that the pipe position is most reliably detected when the threshold value is 7 or 8.

Protective effect of Buddha's Temple extract against tert-butyl hydroperoxide stimulation-induced oxidative stress in DF-1 cells

  • Eun Hye Park;Sung-Jo Kim
    • Animal Bioscience
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    • v.36 no.7
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    • pp.1120-1129
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    • 2023
  • Objective: This study aimed to determine the protective efficacy of Buddha's Temple (BT) extract against tert-butyl hydroperoxide (t-BHP)-induced oxidative stress in Gallus gallus chicken embryo fibroblast cell line (DF-1) and its effects on the cell lipid metabolism. Methods: In this experimental study, Gallus gallus DF-1 fibroblast cells were pretreated with BT 10-7 for 24 hours, followed by their six-hour exposure to t-BHP (100 μM). Water-soluble tetrazolium salt-8 (WST-8) assays were performed, and the growth curve was computed. The intracellular gene expression changes caused by BT extract were confirmed through quantitative polymerase chain reaction (qPCR). Flow cytometry, oil red O staining experiment, and thin-layer chromatography were performed for the detection of intracellular metabolic mechanism changes. Results: The WST-8 assay results showed that the BT pretreatment of Gallus gallus DF-1 fibroblast cell increased their cell survival rate by 1.08%±0.04%, decreased the reactive oxygen species (ROS) level by 0.93%±0.12% even after exposure to oxidants, and stabilized mitochondrial activity by 1.37%±0.36%. In addition, qPCR results confirmed that the gene expression levels of tumor necrosis factor α (TNFα), TIR domain-containing adapter inducing IFN-beta (TICAM1), and glucose-regulated protein 78 (GRP78) were regulated, which contributed to cell stabilization. Thin-layer chromatography and oil red O analyses showed a clear decrease in the contents of lipid metabolites such as triacylglycerol and free fatty acids. Conclusion: In this study, we confirmed that the examined BT extract exerted selective protective effects on Gallus gallus DF-1 fibroblast cells against cell damage caused by t-BHP, which is a strong oxidative inducer. Furthermore, we established that this extract significantly reduced the intracellular ROS accumulation due to oxidative stress, which contributes to an increase in poultry production and higher incomes.

Impact of particulate matter on the morbidity and mortality and its assessment of economic costs

  • Ramazanova, Elmira;Tokazhanov, Galym;Kerimray, Aiymgul;Lee, Woojin
    • Advances in environmental research
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    • v.10 no.1
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    • pp.17-41
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    • 2021
  • Kazakhstan's cities experience high concentrations levels of atmospheric particulate matter (PM), which is well-known for its highly detrimental effect on the human health. A further increase in PM concentrations in the future could lead to a higher air pollution-caused morbidity and mortality, causing an increase in healthcare expenditures by the government. However, to prevent elevated PM concentrations in the future, more stringent standards could be implemented by lowering current maximum allowable PM concentration limit to Organization for Economic Co-operation and Development (OECD)'s limits. Therefore, this study aims to find out what impact this change in environmental policy towards PM has on state economy in the long run. Future PM10 and PM2.5 concentrations were estimated using multiple linear regression based on gross regional product (GRP) and population growth parameters. Dose-response model was based on World Health Organization's approach for the identification of mortality, morbidity and healthcare costs due to air pollution. Analysis of concentrations revealed that only 6 out of 21 cities of Kazakhstan did not exceed the EU limit on PM10 concentration. Changing environmental standards resulted in the 71.7% decrease in mortality and 77% decrease in morbidity cases in all cities compared to the case without changes in environmental policy. Moreover, the cost of morbidity and mortality associated with air pollution decreased by $669 million in 2030 and $2183 million in 2050 in case of implementation of OECD standards. Thus, changing environmental regulations will be beneficial in terms of both of mortality reduction and state budget saving.

Differential Proteomic Analysis of Chinese fir Clone Leaf Response to Salicylic Acid

  • Yang, Mei;Lin, Sizu;Cao, Guangqiu
    • Journal of Forest and Environmental Science
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    • v.26 no.2
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    • pp.83-94
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    • 2010
  • Chinese fir (Latin name: Cunninghaimia lanceolata) is one of the major commercial coniferous trees. Most of Chinese fir forests are managed in successive rotation sites, which lead productivity to decline. Autotoxicity is the important reason for soil degradation of Chinese fir plantation, especially, phenolic acids are considered as the major allelopathic toxins which induce autotoxicity in Chinese fir rotation stands. We performed here proteomic approach to investigate the response of proteins in Chinese fir leaves to salicylic acid. The tube plantlets of Chinese fir clone were treated with 120 mg/L salicylic acid for 1, 3 and 5th day. 2-DE, coupled with MALDI-TOF-TOF/MS, was used to separate and identify the responsive proteins. We found 12, 7, and 12 candidate protein spots that were up- or down-regulated by at least 2.5 fold after 1, 3, and 5th day of the stress, respectively. Of these protein spots, 16 spots were identified successfully. According to the putative physiological functions, these proteins were categorized into five classes (1) the proteins involved in protein stability and folding, including 26S proteome, Grp78, Hsp70, Hsp90 and PPIase; (2) the protein involved in photosynthesis and respiration, including OEC 33 kDa subunit, GAPDH; (3) the protein related to cell endurance to acid, F-ATPase; (4) the protein related to cytoskeleton, tubulin; (5) the protein related to protein translation: prolyl-tRNA synthetase. These results give new insights into autotoxic substance stress response in Chinese fir leaves and provide preliminary footprints for further studies on the molecular signal mechanisms induced by the stress.

Effect of Bcl-2 Inhibitor Treatment on Embryo Developmental Competence, Apoptosis and ER-stress in Pigs (Bcl-2의 저해제 처리에 따른 돼지 수정란의 배발달 능력, 세포 사멸 및 소포체 스트레스 양상)

  • Hong, Joo-Hee;Min, Sung-Hun;Lee, Enok;Son, Hyeong-Hoon;Yeon, Ji-Yeong;Park, Humdai;Koo, Deog-Bon
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.167-172
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    • 2012
  • The key regulators of apoptosis are the interacting protein of the Bcl-2 family. Bcl-2, an important member of this family, blocks cytochrome C release by sequestering pro-apoptotic BH3-only proteins such as Bid, Bad, Bax and Bim. The pro-survival family members (Bcl-2, Bcl-XL, Bcl-W) are critical for cell survival, since loss of any of them causes cell death in certain cell type. However, its role during early porcine embryonic development is not sufficient. In this study, we traced the effects of Bcl-2 inhibitor, ABT-737, on early porcine embryonic development. We also investigated several indicators of developmental potential, including gene expression (apoptosis-related genes) and apoptosis, which are affected by ABT-737. Porcine embryos were cultured in the PZM-3 medium with or without ABT-737 for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without ABT-737 ($14.7{\pm}3.0$ vs $30.3{\pm}4.8%$, p<0.05). TUNEL assay showed that the number of containing fragmented DNA at the blastocyst stage increased in the ABT-737 treated group compared with control (4.7 vs 3.7, p<0.05). The mRNA expression of the pro-apoptotic gene Bax increased in ABT-737 treated group (p<0.05), whereas expressions of the anti-apoptotic Bcl-2 family members (Bcl-2, Bcl-XL, Bcl-W) decreased (p<0.05). Also, expressions of the ER stress indicator genes (GRP78, XBP-1 and sXBP-1) increased in ABT-737 treated group (p<0.05). In conclusion, Bcl-2 is closely associated with of apoptosis- and ER stress-related genes expressions and developmental potential in pig embryos.

Involvement of Endoplasmic Reticulum Stress Response in the Neuronal Differentiation

  • Cho, Yoon-Mi;Jang, Yoon-Seong;Jang, Young-Min;Seo, Jin-Young;Kim, Ho-Shik;Lee, Jeong-Hwa;Jeong, Seong-Whan;Kim, In-Kyung;Kwon, Oh-Joo
    • The Korean Journal of Physiology and Pharmacology
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    • v.11 no.6
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    • pp.239-246
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    • 2007
  • Expressions of endoplasmic reticulum stress response (ERSR) genes were examined during the neuronal differentiation of rat fetal cortical precursor cells (rCPC) and rat pheochromocytoma PC12 cells. When rCPC were differentiated into neuronal cells for 7 days, early stem cell marker, nest in, expression was decreased from day 4, and neuronal markers such as neurofilament-L, -M and Tuj1 were increased after day 4. In this condition, expressions of BIP, ATF6, and phosphorylated PERK as well as their down stream signaling molecules such as CHOP, ATF4, XBP1, GADD34, Nrf2 and $p58^{IPK}$ were significantly increased, suggesting the induction of ERSR during neuronal differentiation of rCPC. ERSR was also induced during the differentiation of PC12 cells for 9 days with NGF. Neurofilament-L transcript was time-dependently increased. Both mRNA and protein levels of Tuj1 were increased after the induction, and the significant increase in NeuN was observed at day 9. Similar to the expression patterns of neuronal markers, BIP/GRP78 and CHOP mRNAs were highly increased at day 9, and ATF4 mRNA was also increased from day 7. These results strongly suggest the induction and possible role of ERSR in neuronal differentiation process. Further study to identify targets responsible for neuronal induction will be necessary.

The Anti-Bacterial Activity and Anti-Inflammatory Effect of Ethanol Complex Extracts of Safflower and Mother Wort (사플라워와 마더워트 등 에탄올복합추출물의 항균활성과 항염증 효과)

  • Hyun Kyoung Kim;Yungi Lee;Subin Choi;DO Wan Kim
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.1
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    • pp.717-724
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    • 2023
  • we are investigated the anti-inflammatory effects of Safflower and Mother wort Ethanol Complex Extracts(SEC) on lipopolysaccharide (LPS)-activated RAW 264.7 cells. The results demonstrated that pretreatment of SEC(500㎍/mL) significantly reduced NO production by suppressing iNOS protein expression in LPS-stimulated cells. Anti-inflammatory effects by Safflower and Mother wort Ethanol Complex Extracts were observed in the following. Safflower and Mother wort Ethanol Complex Extracts inhibited the translocation of NF-κB from the cytosol to the nucleus via the suppression of IκB-α phosphorylation and also inhibited LPS-stimulated NF-κB transcriptional activity. These findings suggest that Safflower and Mother wort Ethanol Complex Extracts exert anti-inflammatory actions and help to elucidate the mechanisms underlying the potential therapeutic values of Safflower and Mother wort Ethanol Complex Extracts. Therefore, Safflower and Mother wort Ethanol Complex Extracts could be regarded as a potential source of natural anti-inflammatory agents.