• Title/Summary/Keyword: Fold

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The development of anti-DR4 single-chain Fv (ScFv) antibody fused to Streptavidin (Streptavidin이 융합된 DR4 항원에 특이적인 single-chain Fv 항체의 개발)

  • Kim, Seo Woo;Wu, Sangwook;Kim, Jin-Kyoo
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.330-342
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    • 2018
  • The Streptavidin and Biotin system has been studied most extensively as the high affinity non-covalent binding of Biotin to STR ($K_D=10^{-14}M$) and four Biotin binding sites in tetrameric Streptavidin makes this system useful for the production of multivalent antibody. For the application of this system, we cloned Streptavidin amplified from Streptomyces avidinii chromosome by PCR and fused to gene of hAY4 single-chain Fv antibody specific to death receptor 4 (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand. The hAY4 single-chain Fv antibody fused to Streptavidin expressed in Escherichia coli showed 43 kDa monomer in heated SDS-PAGE. However, this fusion protein shown in both non-heated SDS-PAGE and Size-exclusion chromatography exhibited 172 kDa as a tetramer suggesting that natural tetramerization of Streptavidin by non-covalent association induced hAY4 single-chain Fv tetramerization. This fusion protein retained a Biotin binding activity similar to natural Streptavidin as shown in Ouchterlony assay and ELISA. Death receptor 4 antigen binding activity of purified hAY4 single-chain Fv fused to Streptavidin was also confirmed by ELISA and Westernblot. In addition, surface plasmon resonance analysis showed 60-fold higher antigen binding affinity of the hAY4-STR than monomeric hAY4 ScFv due to tetramerization. In summary, hAY4 single-chain Fv fused to Streptavidin fusion protein was successfully expressed and purified as a soluble tetramer in E. coli and showed both Biotin and DR4 antigen binding activity suggesting possible production of bifunctional and tetrameric ScFv antibody.

Effect of Semisulcospira libertina Extracts from Different Extraction Processes on Liver Cell Toxicity and Ethanol Metabolism (간세포 독성과 에탄올 대사에서 추출 조건에 따른 다슬기 추출물의 효과)

  • Cho, Kyoung Hwan;Choo, Ho Jin;Seo, Min Gyun;Kim, Jong Cheol;Shin, Yu Jin;Ryu, Gi Hyung;Cho, Hee Young;Jeong, Chi-Young;Hah, Young-Sool
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.158-166
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    • 2017
  • Although Semisulcospira libertina is generally regarded as a supplement for the alleviation of alcohol hangover, little is known about its effects on cell metabolism. Therefore, this study was conducted to analyze the constituents of the extracts prepared using different extraction methods and to compare their biochemical properties. The amino acid contents were found to be much higher in acidic and enzymatic hydrolysates than hot water extracts from S. libertina. DPPH radical scavenging activities in acidic and enzymatic hydrolysates were higher than those of hot water extracts. Three types of S. libertina hydrolysate was added to HepG2 cells damaged by acetaminophen (AAP), after which the survival rate of HepG2 cell were measured. In addition, lactate dehydrogenase (LDH) activities in the culture media were evaluated. The survival rates of HepG2 cells were $77.0{\pm}4.3%$ and $81.5{\pm}1.3%$ at 3 h and 5h enzymatic hydrolysates, respectively. These cell survival rates were higher compared to those of the negative control group ($67.8{\pm}4.3%$) treated only with acetaminophen. Cellular toxicities induced by treatment with AAP were also significantly alleviated in response to treatment with the extracts of S. libertina. In addition, the activities of 2 key enzymes that metabolize ethanol, alcohol dehydrogenase and aldehyde dehydrogenase, were upregulated by 4.7- and 2.7-fold respectively in response to treatment with a 3 h enzymatic hydrolysate of S. libertina. Taken together, these results provide biochemical evidence of the method by which S. libertina exerts its biological functions, including the alleviation of alcohol hangover and the protection of liver cells against toxic insults.

A study on Indoor Insulating Tent Design for Senior Citizens who Live Alone (독거노인을 위한 융복합 실내 보온용 텐트디자인에 관한 연구)

  • Lee, Dae Hyun;Kim, Sang Sik
    • Korea Science and Art Forum
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    • v.37 no.2
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    • pp.219-230
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    • 2019
  • According to the National Statistical Office, the number of senior citizens aged 65 or over has exceeded 7 million as of 2017, and the number of senior citizens who live alone among them exceeded 1.3 million. Most of the senior citizens who live alone suffer from absolute poverty (68.5% of them has monthly income less than 500,000 won) and they have difficulties for basic living in the blind spot of our society. In particular, the heating is quite a serious issue for the senior citizens who live alone and belong to vulnerable social group in the winter, and to make it worse, they are exposed to frequent fire accidents due to the negligence in handling electric appliances such as electric heaters and electric pads. The main reason the indoor tent products are being used by senior citizens who live alone is that it saves energy, ensures warm sleep, and improves fire safety. Following the expansion of the indoor tent market, this study focused on the idea that there is a need for an in door tent suitable for senior citizens who live alone and belong to low income bracket and intended to improve its efficiency in relation to use, resting, and storage. For this, a basic survey was conducted on the products of existing brands to analyze advantages and disadvantages and it was possible to understand the demands that consumers have for existing products. Accordingly, a survey on consumer preference was conducted using a designed model and the Zabara typp, which demonstrated the best efficiency in terms of installation convenience, space usability, and appearance design was selected. Based on the results of selection, the product design and final prototype were completed. The results and details of the study are as follows; First, factors that were not recognized in product development phase could be identified through usability survey and interview with actual users. Second, for the effective aspect of the prototype, senior citizens could install and fold the tend more easily and quickly than expected. Based on these results of this study, it is expected that not only senior citizens who live alone but also various other users can use the tent to create another comfortable private space indoors.

A Study on Classifications and Trends with Convergence Form Characteristics of Architecture in Tall Buildings (초고층빌딩의 융합적 건축형태 분류와 경향에 관한 연구)

  • Park, Sang Jun
    • Korea Science and Art Forum
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    • v.37 no.5
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    • pp.119-133
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    • 2019
  • This study is as skyscrapers are becoming increasingly taller, more constructors have decided the height alone cannot be a sufficient differentiator. As a result, atypical architecture is emerging as a new competitive factor. Also, it can be used for symbolizing the economic competitiveness of a country, city, or business through its form. Before the introduction of digital media, there was a discrepancy between the structure and form of a building and correcting this discrepancy required a separate structural medium. Since the late 1980s, however, digitally-based atypical form development began to be used experimentally, and, until the 2000s, it was used mostly for super-tall skyscrapers for offices or for industrial chimneys and communication towers. Since the 2000s, many global brand hotels and commercial and residential buildings have been built as super-tall skyscrapers, which shows the recent trend in architecture that is moving beyond the traditional limits. Complex atypical structure is formed and the formative characteristics of diagonal lines and curved surfaces, which are characteristics of atypical architecture, are created digitally. Therefore, it's goal is necessary to identify a new relationship between the structure and forms. According to the data of Council on Tall Buildings and Urban Habitat (CTBUH), 100-story and taller buildings were classified into typical, diagonal, curved, and segment types in order to define formative shapes of super-tall skyscrapers and provide a ground of the design process related to the initial formation of the concept. The purpose of this study was to identify the correlation between different forms for building atypical architectural shapes that are complex and diverse. The study results are presented as follows: Firstly, complex function follows convergence form characteristics. Secondly, fold has inside of architecture with repeat. Thirdly, as curve style which has pure twist, helix twist, and spiral twist. The findings in this study can be used as basic data for classifying and predicting trends of the future super-tall skyscrapers.

Global Rice Production, Consumption and Trade: Trends and Future Directions

  • Bhandari, Humnath
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2019.09a
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    • pp.5-5
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    • 2019
  • The objectives of this paper are (i) to analyze past trends and future directions of rice production, consumption and trade across the world and (ii) to discuss emerging challenges and future directions in the global rice industry. Rice is a staple food of over half of the world's 7.7 billion people. It is an important economic, social, political, and cultural commodity in most Asian countries. Rice is the $1^{st}$ most widely consumed, $2^{nd}$ largely produced, and $3^{rd}$ most widely grown food crop in the world. It was cultivated by 144 million farms in over 100 countries with harvested area of over 163 million ha producing about 745 million tons paddy in 2018. About 90% of the total rice is produced in Asia. China and India, the biggest rice producers, account for over half of the world's rice production. Between 1960 and 2018, world rice production increased over threefold from 221 to 745 million tons (2.1% per year) due to area expansion from 120 to 163 million ha (0.5% per year) and paddy yield increase from 1.8 to 4.6 t/ha (1.6% per year). The Green Revolution led massive increase in rice production prevented famines, provided food for millions of people, reduced poverty and hunger, and improved livelihoods of millions of Asians. The future increase in rice production must come from yield increase as the scope for area expansion is limited. Rice is the most widely consumed food crop. The world's average per capita milled rice consumption is 64 kilograms providing 19% of daily calories. Asia accounted for 84% of global consumption followed by Africa (7%), South America (3%), and the Middle East (2%). Asia's per capita rice consumption is 100 kilograms per year providing 28% of daily calories. The global and Asian per capita consumption increased from the 1960s to the 1990s but stable afterward. The per capita rice consumption is expected to decline in Asia but increase outside Asia especially in Africa in the future. The total milled rice consumption was about 490 million tons in 2018 and projected to reach 550 million tons by 2030 and 590 million tons by 2040. Rice is thinly traded in international market because it is a highly protected commodity. Only about 9% of the total production is traded in global rice market. However, the volume of global rice trade has increased over six-fold from 7.5 to 46.5 million tons between the 1960s and 2018. A relatively small number of exporting countries interact with a large number of importing countries. The top five rice exporting countries are India, Thailand, Vietnam, Pakistan, and China accounting for 74% of the global rice export. The top five rice importing countries are China, Philippines, Nigeria, European Union and Saudi Arabia accounting for 26% of the global rice import. Within rice varieties, Japonica rice accounts for the highest share of the global rice trade (about 12%) followed by Basmati rice (about 10%). The high concentration of exports to a few countries makes international rice market vulnerable to supply disruptions in exporting countries, leading to higher world prices of rice. The export price of Thai 5% broken rice increased from 198 US$/ton in 2000 to 421 US$/ton in 2018. The volumes of trade and rice prices in the global market are expected to increase in the future. The major future challenges of the rice industry are increasing demand due to population growth, rising demand in Africa, economic growth and diet diversification, competition for natural resources (land and water), labor scarcity, climate change and natural hazards, poverty and inequality, hunger and malnutrition, urbanization, low income in rice farming, yield saturation, aging of farmers, feminization of agriculture, health and environmental concerns, improving value chains, and shifting donor priorities away from agriculture. At the same time, new opportunities are available due to access to new technologies, increased investment by the private sector, and increased global partnership. More investment in rice research and development is needed to develop and disseminate innovative technologies and practices to overcome problems and ensure food and nutrition security of the future population.

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Comparative Analysis of Detection Methods for Food-borne Pathogens in Fresh-cut Agricultural Materials (신선 농산물내 식중독균 검출 방법의 비교 분석)

  • Jang, Hye-Jeong;Kim, Hye-Jeong;Park, Ji-in;Yu, Sun-Nyoung;Park, Bo-Bae;Ha, Gang-Ja;Ahn, Soon-Cheol;Kim, Dong-Seob
    • Journal of Life Science
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    • v.31 no.1
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    • pp.10-16
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    • 2021
  • The consumption of fresh-cut agricultural materials is increasing due to increased public interest in health and the increase of single-person households. Most fresh-cut agricultural materials can be eaten without heating, thus easily exposing the consumer to food-borne pathogens. As a result, food-borne diseases are increasing worldwide. In the analysis of food-borne pathogens, it is important to detect the strains, but this is time consuming and laborious. Alternative detection methods that have been introduced, include polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), which is performed without prior culturing. Samples of fresh-cut agricultural materials, such as vegetables, were analyzed by the culture-based method. In 129 samples, non-pathogenic Escherichia coli (3.9%), Bacillus cereus (31.8%), Clostridium perfringens (5.4%), Yersinia enterocolitica (0.8%), and enterohemorrhagic E. coli (0.8%) were detected. Eight samples contaminated with bacteria were randomly selected, further analyzed by PCR-DGGE, and compared with the culture-based method. Two cases detected non-pathogenic E. coli by PCR-DGGE only, despite a lack of detection by the culture method. It was supposed there was possibility of sample loss during its 10-fold dilution for appropriate cultivation. In the detection of high-risk food-borne pathogens, it was found that the detection limit was lower in PCR-DGGE than in the culture-based method (10 CFU/g). This suggests that PCR-DGGE can be alternatively used to detect strains. On the other hand, low-risk food-borne pathogens seem to have higher detection limits in PCR-DGGE. Consequentially, this study contributes to the improvement of food-borne pathogen detection and the prevention of its related-diseases in fresh-cut agricultural materials.

Comparative Transcriptome Analysis of the Response of Two Lines of Rapeseed (Brassica napus L.) to Cold Stress (유채 두 계통에서 저온 스트레스에 반응하는 전사체 발현 비교 분석)

  • Lee, Ji-Eun;Kim, Kwang-Soo;Cha, Young-Lok;An, Da-Hee;Byun, Jong-Won;Kang, Yong-Ku
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.1
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    • pp.37-71
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    • 2021
  • Rapeseed is a typical winter crop, and its freezing stress tolerance is a major feature for winter survival. Therefore, it is important to comprehend clearly the physical and molecular mechanisms of rapeseed under freezing stress conditions. This study investigates the physical and transcriptome changes of two rapeseed lines, 'J8634-B-30' and 'EMS26', under cold acclimation and freezing temperature treatments. The proline content of 'J8634-B-30' at 5 ℃ increased 8.7-fold compared to that before treatment, and there was no significant change in that of 'EMS26' RNA-sequencing analysis revealed 5,083 differentially expressed genes (DEGs) of 'J8634-B-30' under cold acclimation condition. Among the genes, 2,784 (54.8%) were up-regulated and 2,299 (45.2%) were down-regulated. The DEGs of 'EMS26' under cold acclimation condition were 5,831 genes, and contained 2,199 up-regulated genes (37.7%) and 3,632 down-regulated genes (62.3%). Among them, only DEGs annotated in the cold response-related signaling pathways were selected, and their expression in the two rapeseed lines was compared. Comparative DEGs analysis indicated that cold response related signaling pathways are proline metabolism and ABA (Abscisic acid) signaling. And ICE (Inducer of CBF expression) - CBF (C-repeat-binding factor) - COR (Cold-regulated) signaling were the significantly differentially expressed transcripts in the two rapeseed lines. The major induced transcripts of 'J8634-B-30' induced P5CS (Δ'-pyrroline-5-carboxylate synthetase), which is related to proline biosynthesis, PYL (pyrabactin resistance-like protein, ABA receptor) and COR413 (cold-regulated 413 plasma membrane 1). In conclusion, these result provide a foundation for understanding the mechanisms of freezing stress tolerance in rapeseeds. Further functional studies should be performed on the freezing stress-related genes identified in this study, which can contribute to the transgenic and molecular breeding for freezing stress tolerance in rapeseed.

Effects of Coenzyme Q10 on the Antioxidant System in Blood and Liver of Laying Hens (산란계에서 Coenzyme Q10 급여가 혈액 및 간 조직의 항산화 작용에 미치는 영향)

  • Moon, Yang-Soo;Jang, In-Surk
    • Korean Journal of Poultry Science
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    • v.49 no.2
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    • pp.115-124
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    • 2022
  • The objective of this study was to investigate the effects of dietary coenzyme Q10 (CoQ10) sources on the antioxidant defense system in the blood and liver of laying hens. Thirty-six 40-wk old Lohmann Brown hens were randomly assigned to three groups based on body weight, with four cages with three layers each. Laying hens were divided into one of the following groups: control (CON), powdered CoQ10 (PCoQ, 100 mg/kg diet), and emulsified CoQ10 (ECoQ, 100 mg/kg diet). All hens were fed a control diet or a control diet supplemented with powdered or emulsified CoQ10 ad libitum for five weeks. There were no differences in body weight, weight gain, and organ weights among the treatment groups, including the liver and spleen. The blood total antioxidant power (TAP) in the ECoQ group increased (P<0.05) by approximately 2-fold compared to that in the CON group. However, there was no significant difference in blood TAP levels between the PCoQ and ECoQ groups, although a decreasing trend (P<0.13) was observed for levels of TAP in the ECoQ group. The mRNA expression and specific activities of superoxide dismutase, glutathione peroxidase, and catalase in the liver were not affected by dietary CoQ10 or type of CoQ10. However, hepatic lipid peroxidation in the ECoQ group was lower (P<0.05) than in the CON group. In conclusion, emulsified CoQ10 increased blood TAP and decreased hepatic lipid peroxidation without affecting antioxidant enzymes, suggesting that emulsified CoQ10 might be more applicable as an active antioxidant supplement than powdered type in laying hens.

Antioxidant and nitric oxide inhibition effect of domestic and foreign fermented black tea extracts (국내외 홍차 추출물의 항산화 효과 및 Nitric Oxide 생성 저해 효과)

  • Moon, Gyo-Ha;Kim, Gyeong-Ji;Lee, Yu-Rim;Kim, Jong Cheol;Shim, Doobo;Chung, Kang-Hyun;Lee, Kwon-Jai;An, Jeung Hee
    • Korean Journal of Food Science and Technology
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    • v.53 no.4
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    • pp.454-462
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    • 2021
  • This study investigated the antioxidant and nitric oxide (NO) inhibition effects of three domestic and seven foreign fermented black tea 70% ethanol extracts. Bosung showed the highest total polyphenol and tannin contents. Jeju showed the highest total flavonoid contents. The theanine content was higher in Hadong (651.50 mg%) than in foreign fermented black tea. At 25 ㎍/mL, the domestic fermented black tea extract showed the highest DPPH and ABTS racial-scavenging activities, reducing power assays. Domestic fermented black tea showed higher NO inhibitor activity than foreign black tea at 50 ㎍/mL. Bosung black tea extract showed an increase in SOD-1 level (1.39-fold) compared to the LPS-only group. Bosung and Jeju decreased the GST protein by 1.52- and 1.46-folds, respectively, compared to the LPS-only group. Thus our results suggest that domestic fermented black tea (Bosung, Jeju, and Hadong) are effective antioxidants in RAW 264.7 cells.

CLK3 is a Novel Negative Regulator of NF-κB Signaling (NF-κB 신호경로에서 CLK3의 새로운 음성 조절자로서의 기능)

  • Byeol-Eun, Jeon;Chan-Seong, Kwon;Ji-Eun, Lee;Ye-Lin, Woo;Sang-Woo, Kim
    • Journal of Life Science
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    • v.32 no.11
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    • pp.833-840
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    • 2022
  • Chronic inflammation has been shown to be closely associated with tumor development and progression. Nuclear factor kappa B (NF-κB) is composed of a family of five transcription factors. NF-κB signaling plays a crucial role in the inflammatory response and is often found to be dysregulated in various types of cancer, making it an attractive target in cancer therapeutics. In this study, CDC-like kinase 3 (CLK3) was identified as a novel kinase that regulates the NF-κB signaling pathway. Our data demonstrate that CLK3 inhibits the canonical and non-canonical NF-κB pathways. Luciferase assays following the transient or stable expression of CLK3 indicated that this kinase inhibited NF-κB activation mediated by Tumor necrosis factor-alpha (TNFα) and Phorbol 12-myristate 13-acetate (PMA), which are known to activate NF-κB signaling via the canonical pathway. Consistent with data on the ectopic expression of CLK3, CLK3 knockdown using shRNA constructs increased NF-κB activity 1.5-fold upon stimulation with TNFα in HEK293 cells compared with the control cells. Additionally, overexpression of CLK3 suppressed the activation of this signaling pathway induced by NF-κB-inducing kinase (NIK) or CD40, which are well-established activators of the non-canonical pathway. To further examine the negative impact of CLK3 on NF-κB signaling, we performed Western blotting following the TNFα treatment to directly identify the molecular components of the NF-κB pathway that are affected by this kinase. Our results revealed that CLK3 mitigated the phosphorylation/activation of transforming growth factor-α-activated kinase 1 (TAK1), inhibitor of NF-κB kinase alpha/beta (IKKα/α), NF-κB p65 (RelA), NF-κB inhibitor alpha (IκBα), and Extracellular signal-regulated kinase 1/2-Mitogen-activated protein kinase (ERK1/2-MAPK), suggesting that CLK3 inhibits both the NF-κB and MAPK signaling activated by TNFα exposure. Further studies are required to elucidate the mechanism by which CLK3 inhibits the canonical and non-canonical NF-κB pathways. Collectively, these findings reveal CLK3 as a novel negative regulator of NF-κB signaling.