• Journal of Life Science
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• v.30 no.8
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• pp.701-707
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• 2020

Gene editing technology using the clustered regularly interspaced short palindromic repeat (CRISPR) and the CRISPR associated protein (Cas)9 has been highly anticipated in developing breeding techniques. In this study, we discuss gene scissors as a tool for silkworm molecular breeding through analysis of Bombyx mori Kynurenine 3-Monooxygenase (BmKMO) gene editing using the CRISPR/Cas9 system and analysis of generational transmission through mutagenesis and selective crossing. The nucleotide sequence of the BmKMO gene was analyzed, and three guide RNAs (gRNAs) were prepared. Each synthesized gRNA was combined with Cas9 protein and then analyzed by T7 endonuclease I after introduction into the BM-N silkworm cell line. To edit the silkworm gene, K1P gRNA and Cas9 complexes were subsequently microinjected into the silkworm embryos; the hatching rate was 18% and the incidence of mutation was 60%. The gene mutation was verified in the heterozygous G0 generation, but no phenotypic change was observed. In homozygotes generated by self-crossing, a mutant phenotype was observed. These results suggest that silkworm molecular breeding using the CRISPR/Cas9 system is possible and could be an effective way of shortening the time required.

• Journal of Life Science
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• v.8 no.5
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• pp.509-519
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• 1998

The effect of N-Nitrosodimethylamine(NDMA) on the glycoconjugates of rat lingual salivary gland was examined by prelectin histochemical methods. Sprague-Dawley rats weighing about 250-300g were divided into control and experimental groups. Each rat of experimental groups was administrated NDMA(17mg/kg) orally and sacrificed in 3, 6, 12, 24, 48, 72, 96 and 120 hours after NDMA administration. The regional differences and change of glycoco-njugates were elucidated by prelectin histochemical methods, such as periodic acid Schiff's(PAS) reaction, alcian blue (AB) pH 2.5, AB pH 0.4, 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 major morphological changes in the von Ebner’s gland of NDMA administrated groups were withering and des-truction of serous acini, diminution and disappearance of cytoplasmic granules and vacuolation in cytoplasm of serous cells, and mucinous changes of duct epithelial cells. These changes were noted in NDMA administrated groups for 12 to 72 hours. In the lingual mucous gland of NDMA administrated groups, the major morphological changes were enlargement, fusion and destruction of mucous acini, loss of cytoplasmic granules and vacuolated generation in cytop-lasm of mucous cells, and mucinous change of duct epithelial cells. These changes were severe in NDMA administra-ted groups for 12 to 72 hours. In NDMA administrated groups of lingual von Ebner's gland for 12 and 72 hours, the neutral glycoconjugates be-come diminished remarkably compared to the control group. The decreased amount of neutral glycoconjugates tended to be gradually recovered from 96 hours group. The acidic glycoconjugates which were not detected in control group were found in a few serous cells of these gland of NDMA administrated groups for 6 to 48 hours and 120 ho-urs. The remarkable decrease of neutral and acidic glycoconjugates was observed in the lingual mucous glands 3, 24 and 48 hours after NDMA administration, and the striking decrease of acidic glycoconjugates was found in 72 hours groups. Among acidic glycoconjugates, sulfated glycoconjugates tended to decrease in NDMA administrated groups for 72 hours, while sialic glycoconjugates were increased in NDMA administrated groups for 3, 12 and 48 hours.

• Journal of Korean Society of Archives and Records Management
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• v.1 no.1
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• pp.37-52
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• 2001

1. establishment and development of China archives science: With the centuries-old history of archives and archives management, early China archives science came into being in 1930s, and the research pushed forward by archives enterprise has made great achievements since then. 1.1 Expanding research fields: Foundation

• Journal of Korean Home Economics Education Association
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• v.26 no.3
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• pp.69-89
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• 2014

The purpose of this paper was to understand the outlines of 1) common and free curricula implemented by the first pilot schools and 2) Home Economics related programs from both curricula. This study analyzed the 42 reports from the first pilot schools which implemented the free learning semester in 2013. The results showed that more than half of schools set two or more emphasis models of the free learning semester. In 15 out of 28 schools, the lesson hours of Home Economics were reduced by one to three hours. Various common curricula were implemented in many aspects of teaching-learning methods, curriculum, evaluation, and subject related career lessons. Free curriculum was implemented in four areas such as career exploration, student optional program, club activity, and art sports activity. In common curriculum, Home Economics was carried out in 17 out of 39 schools. The Home Economics class included subject related career lessons and improvement of teaching- learning methods. Various interesting activities and programs as a free curriculum were implemented in career exploration, students' optional program, and club activities in 38 schools. The most frequently implemented was the activity program related to food and nutrition. Further research was proposed to develop students' optional programs related to Home Economics subject, responding to their needs.

• Journal of Korean Society of Forest Science
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• v.101 no.3
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• pp.387-394
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• 2012

The study was conducted to develop an effective forest resources use models for an alternate use of abandoned quarry by an attitude survey. According to the result of survey, a pessimistic view due to dust, noise pollution, and forest damage was 5% higher than an affirmative view by economic benefits from the development of quarry. The 42% of the respondents preferred the alternate use of abandoned quarry and the 25% of the respondents wanted an art and cultural space. The optimum size of alternate use was 5-10 ha (43%) with the requirement of nearby residents (32%). According to the SWOT analysis for abandoned quarry, the strength factors were an effective use of land, the content development of modern industrial inheritance + cultural and art fusion, attraction for nearby city and visitors, a harmony of beauty landscape and clean environment, and a sustainable increase of domestic and foreign visitors with the 5-day-work week. The opportunity factors were the improvement of traffic networks through KTX and local highway, the creation of the new growth engines with the establishment of artistic creation belts, the providing of unique cultural and art space through grafting of tour and education, the creation of local income through stone processed goods, and the vitalization of local development through eco-city. The weakness factors were a psychological remoteness and backwardness, and the weakness of staying tour infra. The threat factors were a poor financial support for sustainable development in nearby quarry and a modify of legal and institutional system for the alternated use of abandoned quarry. The developed restoration models for the alternate use in abandoned quarry are classified to a sculpture park, a waterfall and lake park, a rock-climbing, a sports park + forest park, a native botanical garden, a culture and art park, a complex park, a water storage site, a water storage site to extinguish forest fire, a geriatric hospital, an agricultural facility, and a school site types etc. The results suggest that the alternate use in the abandoned soil and stone quarry is needed to establish facility use models with consideration of user's preference.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.4
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• pp.389-394
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• 2006

The change of antioxidative character by germination of brown rice was evaluated. From the total methanolic extract of brown rice, 2.5 mm-germinated brown rice, and 5 mm-germinated brown rice, SOD-like activity and nitrite scavenging ability were identified as antioxidative character. SOD-like activities and nitrite scavenging abilities of all samples were changed dose-dependently and germination-dependently. After successive partitioning with hexane, ethyl acetate (EtOAc) and water, each fraction was tested for these activities. SOD-like activities of all fractions were increased by germination, and especially hexane fraction and EtOAc fraction of 5 mm-germinated brown rice had more strong activities than 50 ppm vitamin C. The $EC_{50}$ values of SOD-like activity showed a gradual decrease by germination and that of EtOAc fraction of 5 mm-germinated brown rice was 17 ppm, which was lower concentration than that of 50 ppm vitamin C. The $IC_{50}$ values of nitrite scavenging ability at PH 1.5 also underwent a great decrease by germination and germinated brown rice had the nitrite scavenging ability at lower concentration than brown rice. The results suggest that SOD-like activity and nitrite scavenging ability are thought to be enhanced by the germination effect.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.1
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• pp.35-43
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• 1996

The appearance of the primordial germ cells (PGC's), early gonadogenesis, sex differentiation and sex ratio of the embryo in the viviparous teleost, Ditrema temmincki were investigated by using photomicroscopy. The PGC's were first observed in the fibrous mesenchymal tissue located between the early alimentary tract and the dorsal body wall in the late embryonic development stage. During the period from the hatching to the individual total length (TL) of 4.0 mm the PGC's were evenly distributed in the fibrous mesenchymal tissue between alimentary tract and body wall. But the period of TL 5.0 mm mesenchymal tissue separated from the dorsal body wall, the PGC's moved to the posterior mesenchymal tissue and formed the primitive gonad. During the early gonadogenesis, differentiation of the testis and ovary were distinguished from the arrangement of the germ cells and somatic cells. Gonad of embryo in TL 10.0 mm can be separated into the ovary and testis by external morphology. The testis had a separated form which was consisted with two lobes, and the ovary had a fused form in half-posterior part. In the testicular differentiation of the embryo, histological pattern of the seminiferous tubule appeared when TL of the embryo was to be 25.0 mm, for the seminal vesicle was formed In the individual TL of 30.0mm. The testis of the embryo with TL of 45.0 mm was similar to that of the adult fish in the external and internal structures. In the ovarian differentiation, formation of the ovigerous folds and the ovarian cavity were clearly observed when the TL reached to 30.0 mm. The ovary from the individual with TL of 60.0 mm was differentiated into a similar ovary as seen in the adult fish in the external and internal structure. Right before parturition the total length of the individual was approximately 63.0 mm of which the individual embryo has an ovary containing the oocytes in the chromatin nucleolus stage, or a testis containing the spermatogonia, respectively. And the embryonic sex ratio of female to male was 1.65 : 1. Ditrema temmincki is dioecism and the pattern of sex differentiation is belonged to a differentiation type.

• Journal of Life Science
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• v.18 no.6
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• pp.796-803
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• 2008

Mutations in the APP gene lead to enhanced cleavage by ${\beta}-$ and ${\gamma}-secretase$, and increased $A{\beta}$ formation, which are closely associated with Alzheimer's disease (AD)-like neuropathological changes. Recent studies have shown that exercise training can ameliorate pathogenic phenotypes ($A{\beta}-42$, BDNF, GLUT-1 and HSP70) in experimental models of Alzheimer's disease. Here, we have used NSE/APPsw transgenic mice to investigate directly whether exercise training ameliorates pathogenic phenotypes within Alzheimer's brains. Sixteen weeks of exercise training resulted in a reduction of $A{\beta}-42$ peptides and also facilitated improvement of cognitive function. Furthermore, GLUT -1 and BDNF proteins produced by exercise training may protect brain neurons by inducing the concomitant expression of genes that encode proteins (HSP-70) which suppress stress induced neuron cell damages from APPsw transgenic mice. Thus, the improved cognitive function by exercise training may be mechanistically linked to a reduction of $A{\beta}-42$ peptides, possibly via activation of BDNF, GLUT-1, and HSP-70 proteins. On the basis of the evidences presented in this study, exercise training may represent a practical therapeutic management strategy for human subjects suffering from Alzheimer's disease.

• Journal of Life Science
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• v.17 no.2 s.82
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• pp.198-203
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• 2007

DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.

• Journal of Life Science
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• v.21 no.7
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• pp.1032-1038
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• 2011

Recently identified Ciona intestinalis voltage sensor-containing phosphatase (Ci-VSP) consists of an ion channel-like transmembrane domain (VSD) and a phosphatase-like domain. Ci-VSP senses the change of membrane potential by its VSD and works as a phosphoinositide phosphatase by its phosphatase domain. In this study, we present the construction of His-tagged phosphatase-like domain of Ci-VSP, its recombinant expression and purification, and its enzymatic activity behavior in order to examine the biochemical behavior of phosphatase domain of Ci-VSP without interference. We found that Ci-VSP(248-576)-His can be eluted with an elution buffer containing 25 mM NaCl and 100 mM imidazole during His-tag purification. In addition, we found the proper measurement condition for kinetics study of Ci-VSP(248-576)-His against p-nitrophenyl phosphate (pNPP). We measured the kinetic constant of Ci-VSP(248-576)-His at $37^{\circ}C$, pH 5.0 or 5.5, under 30 min of reaction time, and less than $2.0\;{\mu}g$ of protein amount. With these conditions, we acquired that Ci-VSP(248-576)-His has $K_m$ of $354{\pm}0.143\;{\mu}M$, $V_{max}$ of $0.0607{\pm}0.0137\;{\mu}mol$/min/mg and $k_{cat}$ of $0.359{\pm}0.009751\;min^{-1}$ for pNPP dephosphorylation. Therefore, we produced a pure form of Ci-VSP(248-576)-His, and this showed a higher activity against pNPP. This purified protein will provide the road to a structural investigation on an interesting protein, Ci-VSP.