• Journal of Life Science
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• v.33 no.10
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• pp.808-819
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• 2023

This study was conducted to develop a selection marker for the identification of the Bacillus licheniformis K12 strain in microbial communities. The strain not only demonstrates good growth at moderate temperatures but also contains enzymes that catalyze the decomposition of various polymer materials, such as proteases, amylases, cellulases, lipases, and xylanases. To identify molecular markers appropriate for use in a microbial community, a search was conducted to identify variable gene regions that show considerable genetic mutations, such as recombinase, integration, and transposase sites, as well as phase-related genes. As a result, five areas were identified that have potential as selection markers. The candidate markers were two recombinase sites (BLK1 and BLK2), two integration sites (BLK3 and BLK4), and one phase-related site (BLK5). A PCR analysis performed with different Bacillus species (e.g., B. licheniformis, Bacillus velezensis, Bacillus subtilis, and Bacillus cereus) confirmed that PCR products appeared at specific locations in B. licheniformis: BLK1 in recombinase, BLK2 in recombinase family protein, and BLK3 and BLK4 as site-specific integrations. In addition, BLK1 and BLK3 were identified as good candidate markers via a PCR analysis performed on subspecies of standard B. licheniformis strains. Therefore, the findings suggest that BLK1 can be used as a selection marker for B. licheniformis species and subspecies in the microbiome.

• Journal of Bio-Environment Control
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• v.30 no.1
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• pp.77-84
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• 2021

This study attempted to find a way to maintain the quality of mixing baby wild leaf vegetables with existing baby leaf vegetables in various ratios. The crops for mixing baby leaf vegetables were Peucedanum japoincum Thunberg and Ligularia stenocephala, as wild vegetables, and red romaine, which is widely used in young leafy vegetables. The mixing ratio of red romaine and wild vegetables was red romaine 0: mantilla oil 5: L. stenocephala ratio 5 (R0: P5: L5), red romaine 3.3: P. japoincum 3.3: L. stenocephala ratio 3.3 (R3.3: P3.3: L3.3), red romaine 5: P. japoincum 2.5: L. stenocephala 2.5 (R5: P2.5: L2.5), red romaine 8: P. japoincum 1: L. stenocephala 1 (R8: P1: L1), red romaine 10: P. japoincum 0: L. stenocephala 0 (R10: P0: L0). All treatments were packaged in OTR (oxygen transmittance) 10,000 cc m-2·day-1·atm-1 film and stored for 27 days at 2℃/85% RH conditions. Fresh weight, carbon dioxide, oxygen, and ethylene concentrations of the baby leaf packages were examined approximately every 3 days, and visual quality, chlorophyll content, and chromaticity were examined on the 27th day of storage. The oxygen and carbon dioxide concentration in the packages were affected by the respiration rate of the crop. As the mixing ratio of lettuce, which had a low respiration rate, increased, the oxygen concentration in the packages was higher and the carbon dioxide concentration was lower. Oxygen concentration decreased significantly after 15 days, but was remained above 16%, and on the contrary, carbon dioxide concentration was kept at 1-4% until the 15th, and then gradually increased to 2-5% on the 27th day. The concentration of ethylene was maintained at 3-6 µL·L-1 until the end of storage (27th day). Visual quality score measured at the end of storage was slightly less than 3.0, which is the limit of marketability of all treatments. Although there was no significant difference, the chlorophyll content (SPAD) of red romaine and P. japoincum were most similar with an initial value in R8:P1:1 treatment, and L. stenocephala was higher value in R8:P1:L1 and R5:P2.5:L2.5 treatments at the end of storage. The leaf color (L∗, a∗, b∗, chroma) of the three crops at end of storage compared with the heat map showed the least change in the R5:P2.5:L2.5 and R8:P1:L1 treatments at the end of storage. Among them, R8:P1:L1 treatment maintained the highest chlorophyll content, the second lowest ethylene concentration, and adequate carbon dioxide concentration of 2-3%. Therefore, it is judged that the mixed ratio of red romaine 8: P. japoincum 1: L. stenocephala 1 (R8: P1: L1) is most suitable for the mixed package of baby leaf vegetables of these three crops.

• Korea and Global Affairs
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• v.1 no.2
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• pp.209-242
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• 2017

The purpose of this study is to contribute to enhancing spiritual combat power as the core of intangible combat power in Korean armed forces through analyses and suggestions on Admiral Yi Sun-sin's leadership for four major sweeping victories based upon jointness which effectively integrates tangible and intangible combat power in armed forces to maximize the synergy of fighting power. As our armed forces has improved their military structure in the dimension of hardwares so as to enhance their efficiency, according to the results of analyzing the process to promote the jointness between our armed forces and our allied powers in the dimension of softwares supporting such hardware dimensions, it was necessary to innovate the system for reinforcing future-oriented spiritual combat power as well as all the tasks related to leadership as the core of intangible combat power jointly and harmoniously. In order to derive tasks about the leadership of Korean armed forces in the dimension of softwares which should be combined with military structural reform for strengthening spiritual combat power for national defense, this study selected research questions linked with jointness. That is, (1) what is the core of military leadership in Western advanced countries in the age of jointness? (2) What are the contemporary illuminations or implications of Korean leaderships through Admiral Yi Sun-sin's war history? Then, this study analyzed literature reviews, this author's field interviews in the time of war participation, and leadership war history focusing on Admiral Yi Sun-sin's leadership for four major sweeping victories. According to the results of these analyses, this study extracted (1) the strategic leadership to predict and prepare the future, (2) the leadership of integration to create synergy effects, and (3) the leadership of knowledge to be practiced focusing on combats. In addition, in order to reinforce spiritual combat power based upon jointness, (1) it is necessary to precede in-depth and substantial leadership diagnosis for enhancing jointness. (2) It is necessary to embody national defense reform as well as integration for jointness improvement after scientifically comparing and analyzing the differentiation and integration between the Ministry of National Defense, army-navy-air force leadership centers, and PKO centers. (3) It is necessary to promote the merger and abolition between institutions related to intangibale combat power under the Ministry of National Defense.

• Journal of Life Science
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• v.28 no.7
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• pp.835-840
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• 2018

Quantitative determination of the protein expression levels is one of the most important parts in assessment of the safety of foods derived from genetically modified (GM) crops. Overexpression of AtCYP78A7, a gene encoding cytochrome P450 protein, has been reported to improve tolerance to abiotic stress, such as drought and salt stress, in transgenic rice (Oryza sativa L.). In the present study, an enzyme-linked immunosorbent assay (ELISA) kit for diagnosing AtCYP78A7 protein including AtCYP78A7-specific monoclonal antibody was developed. GST-AtCYP78A7 recombinant protein was induced and purified by affinity column. Four monoclonal antibodies (mAb 6A7, mAb 4C2, mAb 11H6, and mAb 7E8) against recombinant protein were also produced and biotinylated with avidin-HRP. After pairing test using GST-AtCYP78A7 protein and lysate of rice samples, mAb 4C2 and mAb 7E8 were selected as a capture antibody and a detecting antibody, respectively, for ELISA kit. Product test using rice samples indicated that percentages of detected protein in total protein were greater than 0.1% in AtCYP78A7-overexpressing transgenic rice (Line 10B-5 and 18A-4), whereas those in negative control non-transgenic rice (Ilpum and Hwayoung) were less than 0.1%. The ELISA kit developed in this study can be useful for the rapid detection and safety assessment of transgenic rice overexpressing AtCYP78A7.

• Cartoon and Animation Studies
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• s.37
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• pp.393-411
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• 2014

The volume of world market for serious game in year 2015 is expected to be about 9.6 trillion, and the volume of educational serious game market is expected to surpass half of the whole serious game market. In Korea, the development of game for educational purpose has dominated around the education enterprises since late 90s. In 2008, 'Serious Game Forum' was founded led by the Ministry of Culture, Sports, and Tourism with experts from many fields in the society and there were progressing of making policies and plans for potential development of the serious game industry, but the effects were not successful than expected. In 2012, the Ministry of Education, Science, and Technology announced commercialization policy of digital textbook by 2015 and the serious game for educational purpose got attention again. Then, the serious game market became more vigorous with the dispersion of smart devices.13) As a result, the serious games on the smart devices or interlocking between the online and smart devices became an important issue in development rather than the online only serious games. Math field has international competitive power through export in the educational serious game market which takes more than half of the serious game market. Therefore, developing serious game for math education is a good area to raise competitiveness in domestic and international game industries. Moreover, it has no received preferences from students and parents although it has high potential for positive change of individuals and society. The reason is that students recognize it as educational content rather than a game and they avoid it, while parents recognize it as game but not an education. This phenomenon happens because the game elements and educational elements are not properly mixed but focused only on education or emphasized only the fun factors of game when it was developed. Therefore, the purpose of this research is to suggest a direction of developing serious games effectively combining with elementary math for elementary students to get interested in math while playing games. The research will analyze the current elementary math textbooks and find contents which may be combined with the game genre that elementary students enjoy playing these days. This research received advice from serious game developers and math education expert group to reflect the inclination of elementary school students, and respond to the demands from parents and educational institutions, and suggested a direction of developing serious games for effective math education.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.12
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• pp.1568-1575
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• 2008

This study was performed to determine alleviating effects of green tea (GT) on the N-nitrosodimethylamine (NDMA) toxicity to mucins in the rat lingual salivary glands. Sprague-Dawley rats were divided into five groups: untreated (Control), one week-NDMA administrated (NDMA), one week-GT administrated after NDMA for one week (NDMAGT), one week-NDMA administrated with GT (GTNDMA), and one-week NDMA administrated with GT after GT for one week (GTGTNDMA). The mucin properties were elucidated by periodic acid Schiff (PAS) reaction, alcian blue (AB) pH 2.5, AB pH 2.5-PAS, aldehyde fuchsin (AF) pH 1.7-AB pH 2.5, and high iron diamine (HID)-AB pH 2.5 staining. The lingual von Ebner's glands of NDMA group showed some changes such as contraction and destruction of the serous acini, decrease and disappearance of the cytoplasmic granules, vacuolation of cytoplasm, and the mucigenous duct cells. Also, in the lingual mucous glands, enlarged lumens, fused acini, disappearance of granules, vacuolation of cytoplasm, and mucigenous duct cells were observed. All the GT administrated groups had a tendency to recover. GTGTNDMA group recovered almost up to the state of the control group. The lingual von Ebner's glands of NDMA group showed a decrease of neutral mucin and the lingual mucous glands showed a decrease of both neutral and acidic mucins decreased in comparison with control group, although mucous acini secreting strong sulfomucin decreased while those secreting non-sulfomucin (sialomucin) increased. All the GT administrated groups had a tendency to recover. The degree of recovery in the GTNDMA group was stronger than in NDMAGT group and that of the GTGTNDMA group was almost up to the state of the control group. Plus, lingual von Ebner's glands of GTGTNDMA group contained much more neutral mucin than in the control group. In conclusion, it is suggested that NDMA exhibit the toxicity which affects on the mucin properties and which green tea alleviates.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.329-334
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• 2010

Escherichia coli O157:H7 causes hemolytic uremic syndrome and hemorrhagic colitis in humans. The objectives of this study were to produce monoclonal antibody(MAb) against E. coli O157:H7 and to develop an enzyme linked immunosorbent assay(ELISA) for the rapid detection of E. coli O157:H7 in agri-stockbreeding. The characterization of MAb produced from hybridoma cell (HKEC 4G8-5) was validated by ELISA and Western blot. The produced MAb was specific to E. coli O157:H7 and showed weak cross-reaction to Staphylococcus aureus. The detection limit of ELISA based on 4G8-5 MAb was $10^5\;cell/mL$. Although the ELISA could not detect E. coli O157:H7 in the meat and sprout samples inoculated with $1{\times}10^1\;cell$/10 g without enrichment, the same samples after enrichment for 6 hr were confirmed to be positive by ELISA. These results indicated that the ELISA combined with short enrichment (6 hr) is useful tool for rapid screening of E. coli O157:H7 in various samples.

• Journal of Life Science
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• v.28 no.6
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• pp.656-662
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• 2018

Flavin-containing monooxygenases from Corynebacterium (cFMOs) were mutagenized based on homology modeling to develop variants with an enhanced indigoid production capability. The four mutants, F170Y, A210G, A210S, and T326S, which fused to a maltose-binding protein (MBP), were constructed, and their biochemical properties were characterized. Of these, purified MBP-T326S required a higher concentration of exogenous FAD (100 mM) than the wild-type MBP-cFMO for optimal activity and showed a 3.8-fold increase in the $k_{cat}/K_m$ value at $100{\mu}M$ FAD compared to that of MBP-cFMO at $2{\mu}M$ FAD. The indole oxygenase activities of MBP-T326S decreased to 63-77% compared to that of the MBP-cFMO In addition, MBP-T326S displayed a very low level of futile NADPH oxidase activities (21-24%) in the absence of a substrate. Mutant proteins except for T326S displayed similar $K_m$ and increased $k_{cat}/K_m$ values compared to the wild-type. MBP-F170Y and -A210S mutants showed elevated indole oxygenase activity higher than 3.1- and 2.9-fold, respectively, in comparison with MBP-cFMO. When indigoid production was carried out in LB broth with 2.5 g/l of tryptophan, Escherichia coli expressing cFMO produced 684 mg/l of indigo and 104 mg/l of indirubin, while cells harboring T326S produced 1,040 mg/l of indigo and 112 mg/l of indirubin. The results indicate that the production of indigo was 13% higher when compared to a previous report in which an E. coli expressing FMO from Methylophaga produced 920 mg/l of indigo. The protein engineering of cFMO based on homology modeling provided a more rational strategy for developing indigoid-producing strains.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.7 no.1
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• pp.15-27
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• 1974

Since the summer of the year 1972 ulcer disease of common carp, colour carp ana goldfish had suddenly spread widely and caused a serious mortality at fish farms especially around Gim-hae and Yang-san, near Busan. The present study aimed to find out the causative organisms, histopathological changes and the way of treatment, and the results are summarized as follows : Two kinds of pathogenic bacteria, i. e, Chondrococcus columnaris and Aeromonas sp. were isolated from the mucus of the lesion. According to the macroscopic findings, these symptoms began with the hemorrhagic spots under scales which fell out, after the collapse of the dermis, which was followed by lesion to form ulcer, and then the muscle was exposed. The fin was eroded mostly from the distal part, but sometimes from the basal part. Gills showed grey colour, and this part of the tissue collapsed. According to the histopathological findings, the dermis was exposed after the collapsed of the epithelium of the skin, and the necrosis of the musculature occurred, the muscular fibre being destroyed. The epithelial cells of gill tissue proliferated, thus gill filaments were conglutinated and collapsed. Fatty degeneration happened at the liver but the other organs seem to be normal. Treatment with the following mixtures were effective the water temperature of $22\~25^{\circ}C$, but not effective when the temperature was under as low as $13\~15^{\circ}C$. Mixture 1. Aivet (HB-115.HCI) 0.3ppm Malachite green 0.2ppm. Dipterex 0.4ppm Mixture 2. Furanace (P-7138) 0.05ppm Malachite green 0.2ppm Dipterex 0.4ppm When lesion healed, the epithelium, dermis, and the muscular fibre were regenerated.

• Analytical Science and Technology
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• v.31 no.5
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• pp.208-218
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• 2018

Autophagy is a cellular process whereby cytosolic materials or organelles are taken up in a double-membrane vesicle structure known as an autophagosome and transported into a lysosome for degradation. Although autophagy has been studied at the genetic, cellular, or biochemical level, systematic ultrastructural quantitative analysis of autophagosomes during the autophagy process by using transmission electron microscopy (TEM) has not yet been reported. In this study, we performed ultrastructural analysis of autophagosomes in wild-type (WT) mouse embryonic fibroblasts (MEFs) and autophagy essential gene (atg5) knockout (KO) MEFs. First, we performed ultrastructural analysis of autophagosomes in WT MEFs compared to atg5 KO MEFs in basal autophagy or starvation-induced autophagy. Although we observed phagopore, early, late autophagosomes, or autolysosomes in WT MEFs, atg5 KO MEFs had immature autophagosomes that showed incomplete closure. Upon starvation, late autophagosomes accumulated in WT MEFs while the number of immature autophagosomes significantly increased in atg5 KO MEF indicating that atg5 plays an important role in the maturation of autophagosomes. Next, we examined autophagosomes in the cell model expressing polyQ-expanded N-terminal fragment of huntingtin. Our TEM analysis indicates that the number of late autophagosomes was significantly increased in the cells expressing the mutant huntingtin, indicating that improving the fusion of autophagosome with lysosome may be effective to enhance autophagy for the treatment of Huntington's disease. Taken together, the results of our study indicate that ultrastructural and quantitative analysis of autophagosomes using TEM can be applied to various human cellular disease models, and that they will provide an important insight for cellular pathogenesis of human diseases associated with autophagy.