• Title/Summary/Keyword: Specific on-resistance

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Solid-phase PEGylation for Site-Specific Modification of Recombinant Interferon ${\alpha}$-2a : Process Performance, Characterization, and In-vitro Bioactivity (재조합 인터페론 알파-2a의 부위 특이적 수식을 위한 고체상 PEGylation : 공정 성능, 특성화 및 생물학적 활성)

  • Lee, Byung-Kook;Kwon, Jin-Sook;Lee, E.K.
    • KSBB Journal
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    • v.21 no.2
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    • pp.133-139
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    • 2006
  • In 'solid-phase' PEGylation, the conjugation reaction occurs as the proteins are attached to a solid matrix, and thus it can have distinct advantages over the conventional, solution-phase process. We report a case study: rhIFN-${\alpha}$-2a was first adsorbed to cation exchange resin and then N-terminally PEGylated by aldehyde mPEG of 5, 10, and 20 kD through reductive alkylation. After the PEGylation, salt gradient elution efficiently recovered the mono-PEGylate in a purified form from the unwanted species such as unmodified IFN, unreacted PEG, and others. The mono-PEGylation and its purification were integrated in a single chromatographic step. Depending on the molecular weight of the mPEG aldehyde used, the mono-PEGylation yield ranged 50-64%. We could overcome the major problems of random, or uncontrollable, multi-PEGylation and the post-PEGylation purification difficulties associated with the solution-phase process. N-terminal sequencing and MALDI-TOF MS confirmed that a PEG molecule was conjugated only to the N-terminus. Compared with the unmodified IFN, the mono-PEGylate showed the reduced anti-viral activity as measured by the cell proliferation assay. The bioactivity was reduced more as the higher molecular weight PEG was conjugated. Immunoreactivity, evaluated indirectly by antibody binding activity using a surface plasmon resonance biosensor, also decreased. Nevertheless, trypsin resistance as well as thermal stability was considerably improved.

Characteristics of LSC coated Metallic Interconnect for Solid Oxide Fuel Cell (LSC가 코팅된 고체산화물 연료전지용 금속연결재의 특성 연구)

  • Pyo, Seong-Soo;Lee, Seung-Bok;Lim, Tak-Hyoung;Park, Seok-Joo;Song, Rak-Hyun;Shin, Dong-Ryul
    • Korean Chemical Engineering Research
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    • v.48 no.2
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    • pp.172-177
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    • 2010
  • This study reports the high-temperature oxidation kinetics, ASR(area specific resistance), and interfacial microstructure of metallic interconnects coated with conductive oxides in oxidation atmosphere at $800^{\circ}C$, The conductive material LSC($La_{0.8}Sr_{0.2}CoO_3$, prepared by Solid State Reaction) was coated on the Crofer22APU. The contact behavior of coating layer/metal substrate was increased by sandblast. The electrical conductivity of the LSC coated Crpfer22APU was measured by a DC two probe four wire method for 4000hr, in air at $800^{\circ}C$. Microstructure and composition of the coated layer interface were investigated by SEM/EDS. These results show that a coated LSC layer prevents the formation and growth of oxide scale such as $Cr_2O_3$ and enhances the long-term stability and electrical performance of metallic interconnects for SOFCs.

ssc-miR-185 targets cell division cycle 42 and promotes the proliferation of intestinal porcine epithelial cell

  • Wang, Wei;Wang, Pengfei;Xie, Kaihui;Luo, Ruirui;Gao, Xiaoli;Yan, Zunqiang;Huang, Xiaoyu;Yang, Qiaoli;Gun, Shuangbao
    • Animal Bioscience
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    • v.34 no.5
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    • pp.801-810
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    • 2021
  • Objective: microRNAs (miRNAs) can play a role in a variety of physiological and pathological processes, and their role is achieved by regulating the expression of target genes. Our previous high-throughput sequencing found that ssc-miR-185 plays an important regulatory role in piglet diarrhea, but its specific target genes and functions in intestinal porcine epithelial cell (IPEC-J2) are still unclear. We intended to verify the target relationship between porcine miR-185 and cell division cycle 42 (CDC42) gene in IPEC-J2 and to explore the effect of miR-185 on the proliferation of IPEC-J2 cells. Methods: The TargetScan, miRDB, and miRanda software were used to predict the target genes of porcine miR-185, and CDC42 was selected as a candidate target gene. The CDC42-3' UTR-wild type (WT) and CDC42-3'UTR-mutant type (MUT) segments were successfully cloned into pmirGLO luciferase vector, and the luciferase activity was detected after co-transfection with miR-185 mimics and pmirGLO-CDC42-3'UTR. The expression level of CDC42 was analyzed using quantitative polymerase chain reaction and Western blot. The proliferation of IPEC-J2 was detected using cell counting kit-8 (CCK-8), methylthiazolyldiphenyl-tetrazolium bromide (MTT), and 5-ethynyl-2'-deoxyuridine (EdU) assays. Results: Double enzyme digestion and sequencing confirmed that CDC42-3'UTR-WT and CDC42-3'UTR-MUT were successfully cloned into pmirGLO luciferase reporter vector, and the luciferase activity was significantly reduced after co-transfection with miR-185 mimics and CDC42-3'UTR-WT. Further we found that the mRNA and protein expression level of CDC42 were down-regulated after transfection with miR-185 mimics, while the opposite trend was observed after transfection with miR-185 inhibitor (p<0.01). In addition, the CCK-8, MTT, and EdU results demonstrated that miR-185 promotes IPEC-J2 cells proliferation by targeting CDC42. Conclusion: These findings indicate that porcine miR-185 can directly target CDC42 and promote the proliferation of IPEC-J2 cells. However, the detailed regulatory mechanism of miR-185/CDC42 axis in piglets' resistance to diarrhea is yet to be elucidated in further investigation.

Trait Variation and Molecular Characterization of Ds insertional rice lines (벼 Ds 삽입변이 계통의 특성변이 및 분자생물학적 특성)

  • Ahn, Byung-Ohg;Kang, Kyung-Ho;Eun, Moo-Yong;Jeon, Yong-Hee;Yun, Doh-Won;Ji, Hyeon-So;Park, Sung-Han;Nam, Min-Hee;Suh, Seok-Cheol;Lee, Myung-Chul
    • Korean Journal of Breeding Science
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    • v.40 no.1
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    • pp.39-47
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    • 2008
  • The 1,874 rice lines were selected from 3,000 Ds insertional mutant pool by Basta herbicide treatment and were surveyed for trait variation and molecular characteristics of genes knocked out by Ds insertion. Compared with "Donjin", an original japonica cultivar used for transformation, Ds insertion mutant pool showed large variation in major agronomic traits including tiller, panicle, and heading etc. Southern blot analysis demonstrated that these lines on the average had two Ds copies in Donjin genome, resulting in 38.4% of one copy, 32.5% of two copies, 16.7% of three copies, and 11.3% of over four copies. GUS analysis showed that 3.9% of lines (73/1,860) had tissue-specific expression in leaves, nodal parts, floral organs such as stigma and pollen, and roots. Data set obtained from agricultural trait variation and molecular characteristics for individual Ds insertional lines would provide researchers with more information for understanding the function of unknown rice genes controlling economically important traits.

Techno-Economic Analysis of Reversible Solid Oxide Fuel Cell System Couple with Waste Steam (폐스팀을 이용한 가역 고체산화물 연료전지의 기술적 경제적 해석)

  • GIAP, VAN-TIEN;LEE, YOUNG DUK;KIM, YOUNG SANG;AHN, KOOK YOUNG
    • Transactions of the Korean hydrogen and new energy society
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    • v.30 no.1
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    • pp.21-28
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    • 2019
  • Reversible solid oxide fuel cell (ReSOC) system was integrated with waste steam for electrical energy storage in distributed energy storage application. Waste steam was utilized as external heat in SOEC mode for higher hydrogen production efficiency. Three system configurations were analyzed to evaluate techno-economic performance. The first system is a simple configuration to minimize the cost of balance of plant. The second system is the more complicated configuration with heat recovery steam generator (HRSG). The third system is featured with HRSG and fuel recirculation by blower. Lumped models were used for system performance analyses. The ReSOC stack was characterized by applying area specific resistance value at fixed operating pressure and temperature. In economical assessment, the levelized costs of energy storage (LCOS) were calculated for three system configurations based on capital investment. The system lifetime was assumed 20 years with ReSOC stack replaced every 5 years, inflation rate of 2%, and capacity factor of 80%. The results showed that the exergy round-trip efficiency of system 1, 2, 3 were 47.9%, 48.8%, and 52.8% respectively. The high round-trip efficiency of third system compared to others is attributed to the remarkable reduction in steam requirement and hydrogen compression power owning to fuel recirculation. The result from economic calculation showed that the LCOS values of system 1, 2, 3 were 3.46 ¢/kWh, 3.43 ¢/kWh, and 3.14 ¢/kWh, respectively. Even though the systems 2 and 3 have expensive HRSG, they showed higher round-trip efficiencies and significant reduction in boiler and hydrogen compressor cost.

Development of Low-Cost, Double-Speed, High-Precision Operation Control System for Range Extender Engine (레인지 익스텐더 전기자동차 엔진용 저가형 2단속도 고정밀 운전제어시스템 개발)

  • Ham, Yun-Young;Lee, Jeong-Jun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.11
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    • pp.529-535
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    • 2018
  • The range extender vehicle runs on a mechanism that allows the small power generation engine to start in the most efficient specific operating range to charge the battery and extend the mileage. In this study, we developed a step motor type intake air supply system that replaces existing throttle body system to develop a simple low cost control logic system. The system was applied to the existing base engine, and in order to improve the performance by increasing the amount of intake air, the effect of changing the length of the intake and exhaust manifold was experimentally examined. As a result, the Type B intake air control actuator operated by one step motor showed higher performance than the Type A in all the operation region, but the performance was lower than that of the base engine due to the increase of flow resistance. To improve this, it was confirmed that the engine performance was improved at both speeds of 2200rpm and 4300rpm when the 140mm adapter was installed in the intake manifold and when the newly designed 70mm exhaust manifold was applied. Through this process, high - precision operation control was realized by connecting the generator load to the optimized engine for the range extender electric vehicle. Experimental results showed that the speed change rate was within ${\pm}2.5%$ at 2200rpm in 1st stage and 4300rpm in 2nd stage and the speed follow-up result of 610 rpm/s was obtained when the speed was increased from 2200rpm to 4300rpm.

Expression of Antimicrobial Peptide (AMP), Moricin Using SUMO Fusion Tag in Escherichia coli (대장균에서 SUMO fusion tag을 이용하여 항균펩타이드인 moricin의 발현)

  • Ahn, Dong-gyu;Park, Sun Ill;Kim, Soon Young
    • Journal of Life Science
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    • v.32 no.12
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    • pp.956-961
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    • 2022
  • Plant Chloroplast have several advantages as an expression platform of biopharmaceuticals over conventional expression platforms such as mammalian cells, yeast and bacteria. First, plants do not serve as a host for mammalian infectious virus and have endotoxin like bacteria which can cause anaphylactic shock. In addition, high copy number of chloroplast genome allows for chloroplast transformants to reach the high level of expression of heterologous genes. Moreover, the integration of transgenes into specific region of chloroplast genomes makes chloroplast transformants unaffected by positional effect which can be frequently observed from nuclear transformants, resulting in loss of transgene expressions. Antimicrobial peptides (AMPs) are a kind of innate immunity which is found from bacteria to humans. Unlike conventional antibiotics, very less dosage of AMPs can have catastrophic effect on bacterial survival. Further, the repeated use of AMPs does not trigger the development of bacterial resistance. Moricin, one of the AMPs, was isolated from Bombyx mori, a silkworm moth. The C-terminal of moricin consists largely of basic amino acids, and the N-terminal has an α-helix structure. Moricin was chosen and expressed in a SUMO/SUMOase without leaving any unwanted amino acids which could potentially affect the anti-bacterial activity of the moricin. The transformation vector used in this study has already been created in this lab for the expression in both prokaryotic systems such as E. coli and chloroplast. The expressed moricin was purified using Ni columns and SUMOase, and the antibacterial activity of the purified moricin was confirmed using an agar diffusion assay.

Application of Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry (Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry의 활용)

  • Pil Seung KWON
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.4
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    • pp.244-252
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    • 2023
  • The timeliness and accuracy of test results are crucial factors for clinicians to decide and promptly administer effective and targeted antimicrobial therapy, especially in life-threatening infections or when vital organs and functions, such as sight, are at risk. Further research is needed to refine and optimize matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based assays to obtain accurate and reliable results in the shortest time possible. MALDI-TOF MS-based bacterial identification focuses primarily on techniques for isolating and purifying pathogens from clinical samples, the expansion of spectral libraries, and the upgrading of software. As technology advances, many MALDI-based microbial identification databases and systems have been licensed and put into clinical use. Nevertheless, it is still necessary to develop MALDI-TOF MS-based antimicrobial-resistance analysis for comprehensive clinical microbiology characterization. The important applications of MALDI-TOF MS in clinical research include specific application categories, common analytes, main methods, limitations, and solutions. In order to utilize clinical microbiology laboratories, it is essential to secure expertise through education and training of clinical laboratory scientists, and database construction and experience must be maximized. In the future, MALDI-TOF mass spectrometry is expected to be applied in various fields through the use of more powerful databases.

Antimicrobial Photodynamic Therapy on Pseudomonas aeruginosa Using a Diode Laser and PhotoMed, Methyl Pheophorbide A, or Radachlorin® (다이오드 레이저와 PhotoMed, Methyl Pheophorbide A, Radachlorin®을 이용한 녹농균에 대한 항균 광역학 요법)

  • Young-Kyu SONG;Keun-Dol YOOK;Ji-Won KIM
    • Korean Journal of Clinical Laboratory Science
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    • v.56 no.1
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    • pp.52-58
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    • 2024
  • Photodynamic therapy (PDT) activates intracellular oxygen using a photosensitizer activated by light of a specific wavelength and is a potential means of treating wound infections caused by antibiotic-resistant bacteria. Pseudomonas aeruginosa (P. aeruginosa) is typically non-pathogenic in healthy individuals but can induce severe illnesses like sepsis in the immunocompromised. Antibiotics have been conventionally used to treat P. aeruginosa infections, but increasing antibiotic resistance caused by drug misuse poses a growing challenge to the management of these infections. This study aimed to investigate the ability of PDT using photosensitizers (PhotoMed, Methyl pheophorbide A, or Radachlorin®) and a diode laser to inhibit P. aeruginosa. Suspensions of P. aeruginosa and a photosensitizer were inoculated into Petri dishes and incubated for 30 minutes. Samples were then irradiated with the laser at 3 J/cm2, and after incubation, colony areas were measured. P. aeruginosa killing rates were 79.65% for PhotoMed, 47.36% for Methyl pheophorbide A, and 40.91% for Radachlorin®. This study shows that PDT using a diode laser and a photosensitizer constitutes an effective practical therapeutic approach for inhibiting P. aeruginosa.

Effects of Controlling the Pine Needie Gall Midaes by Salicylic Acid Content in Needles of Some Pinus spp. (소나무류(類) 침엽내(針葉內)의 salicylic acid에 의한 솔잎혹파리의 방제효과(防除效果))

  • Son, Doo-Sik;Eom, Tae-Jin;Choi, Chang-Ok;Zhang, Ruo Ming
    • Journal of Korean Society of Forest Science
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    • v.88 no.1
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    • pp.31-37
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    • 1999
  • The frequency of gall formation by pine needle gall midges ranged from 35% to 40% in both P. densiflora and P. thunbergii species. However, there was no indication of gall development from larva on both P. virgiana and P. rigida species suggesting that some specific compounds might play role in preventing gall formation of larva. The susceptible species to pine needle gall midges including P. densiflora and P. thunbergii contained a little salicylic acid under the free phenolic compound conditions while the resistant species against such insects including P. virginiana and P. rigida species contained about 37ppm to 50ppm of salicylic acid. Thus, this compound might have important roles in insect resistance. The contents of internal salicylic acid in the needles of susceptible pines increased from 9.5ppm to 20.6ppm after direct external irrigation of salicylic acid solution and flour treatment on roots. As a result, the frequency of gall formation decreased dramatically 17~19 times lower when compared with that of control. According to our results, the application of salicylic acid for effective prevention against insect damages should be performed before hatching eggs. The frequency of gall formation in the hybrid pines of P. thunbergii(susceptible) and P. virginiana(resistant) showed approximately 24% as average value of those in both species. In the case of specific individuals, it ranged from 2.8% to 11.5% in hybrids of both species. Therefore, effective production of insect resistant pines can be obtained through hybrids by crossing between susceptible and resistant species including P. thunbergii and P. virginiana by increasing internal salicylic acid contents of the needles.

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