• 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.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.2 no.2
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• pp.53-68
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• 1997

Lake Shihwa, artificially constructed since 1988, shows a typical two-layered system depending on strong haline density stratification. Sill of the water gate at 6 m depth greatly restricts physical mixing with outer seawater and circulation in the lake, and contributes to the enhancement of anoxic environment in the deeper layer. With this enclosed physical environment, Lake Shihwa receives enormous amounts of organics, ammonia, and other pollutants from the neighboring municipal and industrial complexes through six major streams, thus developing biogeochemical differentiation of anoxic to suboxic environment in the high saline bottom water and highly eutrophicated brackish surface water. This study investigated vertical structures, biogeochemical behaviors and processes of various organic and inorganic compounds around oxic-anoxic interface. Nitrite and nitrate rapidly decreased below the pycnocline where about $1{\times}10^8$ tons of hypoxic bottom water exist. In this bottom layer, ammonium ranged from 75 to 360 ${\mu}M$ mainly resulting from deamination of dissolved organic nitrogen and ammonification of precipitated organic particles. Despite large amounts of surface water discharge and dilution by outer seawater inflow about $3{\times}10^8$ tons from April to August, 1996, bottom layer did not show any improvement of water quality and maintained highly reduced environment. The main reason seems to be imbalance between ineffectiveness of dilution due to shallow depth and large surface area, overloaded POC influx from the eutrophicated surface biological activity, and poor replenishment of oxygen in this artificial lake system. Therefore, as long as current salinity dependent two-layered system maintains with its physical limitations, any improvement of water quality cannot be foreseen in Lake Shihwa.

• Korean Journal of Plant Resources
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• v.19 no.2
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• pp.300-307
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• 2006

To clarify the morphological and geographical differentiation among the polyploid complexes of L. lancifolium collections in Korea, the mo게hological variation of 173 accessions were analyzed by ANOVA (one-way analysis of variance) and PCA (principal component analysis) on the basis of 38 morphological characters. 173m accessions were grouped into 78 diploids and 95 triploids by ploid levels and the triploids separated into 75 inland triploids (all around the Korea) and 20 island triploids (Backryung-do and Sochung-do, westemmost and northernmost islands of Korea) by geographic distribution and morphology. Island triploids showed significant morphological differences with inland triploids in ANOVA by many floral and leaf characters. In PCAs, diploids were separated from inland triploids by having longer plant height, smaller flower characters, higher pollen fertility and more stomata. The first four principal components accounted for 44.1% of the total variation. Plots of the island and inland groups for the first and second principal components separated each other with slight overlapping. Although the ploid forms are different between diploid and island triploid, island triploids were more closely overlapped with diploids by principal component 1 and 2 than inland triploids. This reflects that the whole external morphology of island triploids are similar to that of diploids. This, the phenotypic differentiation between inland and island triploids seems to be partly related to their geographical origins.

• The Journal of the Petrological Society of Korea
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• v.10 no.2
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• pp.106-120
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• 2001

The Busan area in the northeastern part of the Ogcheon metamorphic zone, Korea, consists mainly of Precambrian Busan and Bakdallyeong gneiss complexes, Ogcheon metamorphic rocks and Mesozoic granitoids. The Busan gneiss complex shows Heart-shaped distribution laying down eastward, and is surrounded by the Ogcheon metamorphic rocks in the central part of the Busan area. In this study structural examination on the main constituent rocks (Busan gneiss complex and Ogcheon metamorphic rocks) was conducted to clarify the geological structure of the Ogcheon metamorphic zone in the Busan area. It indicates that the geological structure was formed at least by three phases of deformation. (1) Dl deformation: the formative period of the structural units of WNW trend (Sanjeoteo, Busan-II, Busan-I, and Chungiu nappes) and the mylonitic foliations related to the compression of NNE-SSW direction, (2) D2 deformation: the differential E-W shortening and N-S extension period of the structural units of WNW trend related to the compression of E-W or WNW-ESE directions, (3) D3 deformation: the formative period of the kink or open folds of E-W trend related to the compression of N-S direction in the eastern and southern parts of Busan area where the structural units of N-S or NNE trends reoriented owing to the intense D2 deformation were developed. These three phases of deformation are closely connected with the distribution of the structural units and the Heart-shaped Busan gneiss complex laying down eastward, and in this paper a new geodynamic model to the Heart-shaped Busan gneiss complex is suggested: Dl deformation-the zonal distribution of WNW trend with a constant width, D2 deformation - the formation of convex wedges northeastward and southward, D3 deformation - the formation of concave wedge westward.

• Economic and Environmental Geology
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• v.28 no.4
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• pp.337-350
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• 1995

The Hwacheon-ri mineralized area is located within the Cretaceous Gyeongsang Basin of the Korean peninsula. The mineralized area includes the Hwacheon, Daeweon, Kuryong and Cheongryong mines. Each of these mines occurs along copper-bearing hydrothermal quartz veins that crosscut late Cretaceous volcanic rocks, although some disseminated ores in host rocks also exist locally. Mineralization can be separated into three distinct stages (I, II, and III) which developed along preexisting fracture zones. Stage I is ore-bearing, whereas stages II and III are barren. The main phase of ore mineralization, stage I, can be classified into three substages (Ia, Ib and Ic) based on ore mineral assemblages and textures. Substage Ia is characterized by pyrite-arsenopyrite-molybdenite-pyrrhotite assemblage and is most common at the Hwacheon deposit. Substage Ib is represented by main precipitation of Cu, Zn, and Pb minerals. Substage Ic is characteristic of hematite occurrence and is shown only at the Kuryong and Cheongryong deposits. Some differences in the ore mineralization at each mine in the area suggest that the evolution of hydrothermal fluids in the area varied in space (both vertically and horizontally) with respect to igneous rocks relating the ore mineralization. Fluid inclusion data show that stage I ore mineralization mainly occurred at temperatures between ${\approx}350^{\circ}$ and ${\approx}200^{\circ}C$ from fluids with salinities between 9.2 and 0.5 wt.% eq. NaCl. In the waning period of substage Ia, the high temperature and salinity fluid gave way to progressively cooler, more dilute fluids of later substage Ib and Ic (down to $200^{\circ}C$, 0 wt.% NaCl). There is a systematic decrease in the calculated ${\delta}^{18}O_{H2O}$ values with paragenetic time in the Hwacheon-ri hydrothermal system from values of ${\approx}2.7$‰ for substage Ia, through ${\approx}-2.8$‰ for substage Ib, to ${\approx}-9.9$‰ for substage Ic. The ${\delta}D$ values of fluid inclusion water also decrease with decreasing temperature (except for the Daeweon deposit) from -62‰ (substage Ia) to -80‰ (substage Ic and stage III). These trends are interpreted to indicate the progressive cooler, more oxidizing unexchanged meteoric water inundation of an initial hydrothermal system which is composed of highly exchanged meteoric water. Equilibrium thermodynamic interpretation of the mineral assemblages with the variation in amounts of chalcopyrite through the paragenetic time, and the evolution of the Hwacheon-ri hydrothermal fluids indicate that the solubility of copper chloride complexes in the hydrothermal system was mainly controlled by the variation of temperature and $fo_2$ conditions.

• Applied Microscopy
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• v.38 no.3
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• pp.175-183
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• 2008

Liver regeneration is a result of highly coordinated proliferation of hepatocytes and non-parenchymal liver cells. At this process, induction of metallothionein (MT), which is low molecular and cysteine rich, has been reported. The present study was carried to find the ultrastructure of hepatocytes and determine the expression of MT in regenerating rat liver after partial hepatectomy. As a result, the remnant liver after PH grew fast from 1 day until 7 days. Various changes were morphologically observed. Disintegration of cell plates and liver lobule appeared shortly after PH. And hepatocytes showed the rapid proliferation, characterized by high nuclear cytoplasmic ratio, weak intercellular junctional complexes, chromatin condensation, increase of ribosomes and mitochondria, and temporary increase of lipid droplets. Finally, remodeling of the liver lobule was completed through the rearrangement of blood vessels and cell plates by 7 days after PH. On histochemistry, immunoreactivity indicating the presence of MT appeared moderately throughout the cytoplasm of control rat hepatocyte. After PH, positive reactions for MT increased at the cytoplasm and the nucleus. These results suggest that the remnant liver cells immediately entered cell proliferation and increase of MT expression after PH. It is thought that MT protein might be associated with transfer of some factors needed to cell division from the cytoplasm to the nucleus for regeneration of the liver after PH.

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.265-273
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• 2004

This study was carried out to examine the effects of fertilization media and sperm concentration on in vitro fertilization (IVF) and development (IVD) of porcine oocytes matured in vitro. Cumulus-oocyte complexes (COCs) were collected from antral follicles of porcine ovaries collected from abattoir, and were matured in vitro in modified NCSU-23 (mNCSU-23) supplemented with 10％ porcine follicular fluid (pFF). After the fertilization by experimental scheme, putative embryos were developed in vitro in NCSU-23. The results are as follows. When the oocytes were fertilized in vitro in modified TBM or modified TLP-PVA by 1 ${\times}$10$^{5}$ sperm/$m\ell$, all of the fertilization parameters were not significantly different between two media. Subsequently, as these putative embryos were developed in vitro in NCSU-23, the percentage of oocytes cleaved and of blastocysts were not different between two media, either. When the oocytes were fertilized in vitro in mTBM by 5${\times}$10$^4$, 1${\times}$10$^{5}$ or 5${\times}$10$^{5}$ sperm/$m\ell$, all of the fertilization parameters were significantly (P<0.05 or P<0.01) increased as sperm concentration was elevated. Subsequently, as these putative embryos were developed in vitro in NCSU-23, the percentage of oocytes cleaved and of blastocysts were significantly boosted (P<0.01) as sperm concentration at fertilization was elevated from 5${\times}$10$^4$ to 1${\times}$10$^{5}$ sperm/$m\ell$, but were not different between 1${\times}$10$^{5}$ and 5${\times}$10$^{5}$ sperm/$m\ell$.

• Journal of Embryo Transfer
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• v.22 no.1
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• pp.1-7
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• 2007

This study was conducted to examine the role of intact cumulus cells during in vitro fertilization (IVF) on sperm penetration, male pronuclear (MPN) formation and subsequent embryo development of oocytes matured and fertilized in vitro. Cumulus-oocyte complexes obtained from the slaughtered gilt ovaries were matured for 44 h in TCM199 containing 10% porcine follicular fluid, epidermal growth factor and hormones. After maturation culture, denuded oocytes or oocytes with intact cumulus cells were coincubated with frozen-thawed boar semen for 8h in a modified tris-buffered medium containing 5mM caffeine and 10mM calcium chloride. Putative zygotes were fixed and examined for sperm penetration and MPN formation (Experiments $1{\sim}3$), or cultured in North Carolina State University-23 medium fo. 156 h (Experiment 3). In Experiment 1, sperm penetration was examined after insemination of denuded oocytes and oocytes with intact cumulus cells at the concentration of $7.5{\times}10^5$ sperm/ml. Optimal sperm concentration for IVF of cumulus-intact oocytes was determined in Experiment 2 by inseminating intact oocytes with $2{\sim}5{\times}10^6$ sperm/ml. In Experiment 3, denuded or intact oocytes were inseminated at the concentrations of $7.5{\times}10^5$ and $4.0{\times}10^6$ sperm/ml, respectively, and in vitro embryo development was compared. Sperm penetration was significantly (p<0.01) decreased in cumulus-intact oocytes compared to denuded oocytes (35.2% vs. 77.4%). Based on the rates of sperm penetration and normal fertilization, the concentration of $4.0{\times}10^6$ sperm/ml was optimal for the IVF of intact oocytes compared to other sperm concentrations. The presence of intact cumulus cells during IVF significantly (p<0.05) improved embryo cleavage (48.8% vs. 58.9%), blastocyst (BL) formation (11.0% vs. 22.8%) and embryo cell number $(22{\pm}2\;vs.\;29{\pm}2\;cells)$ compared to denuded oocytes. In conclusion, these results suggest that intact cumulus cells during IVF inhibit sperm penetration but improve embryo cleavage, BL formation and embryo cell number of porcine embryos produced in vitro.

• The Journal of Korean Society of Virology
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• v.27 no.2
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• pp.227-237
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• 1997

In order to search for anti-Herpes simplex virus (HSV) type-1 agents from Korean medicinal herbs and Korean traditional prescriptions (herb complexes), we selected 80 medicinal herbs and 45 prescriptions, based on a review of the Korean traditional medicine books. Both methanol extracts and boiling-water extracts were tested by means of the MTT assay (tetrazolium based colorimetric assay). Ten of the 125 methanol extracts: CM-11, CM-18, CM-19, CM-21, CM-22, CM-39, MM-3, MM-18, MM-29, MM-73 (see explanation of nomenclature below), showed efficacy against HSV-1. Twelve of the water extracts: CW-2, CW-3-I, CW-3-II, CW-18, MW-3, MW-5 MW-6, MW-12, MW-47, MW-69, MW-73 and MW-79 were active. #3 (individual herb) and #73 (individual herb) were interesting because both water and methanol extracts were active. Especially, #3 is a part of composition of Hong-il-$laksamd{\check{u}}ngbang$ and Hojanghaedokt'ang which have anti-HSV-1 activitives. The SI value of MW-69 and CW-18 was relative high as $10.2{\pm}0.7$ and $11.8{\pm}2.2$. The cytotoxic effect on Vera cells of $Panch'{\check{o}}nch'onch'{\check{o}}ngbang$, Taraxacum platycarpum H. Dahlst. and acycloguanosine was determined by MTT assay. Water extracts of $Panch'{\check{o}}nch'onch'{\check{o}}ngbang$ (prescription) and Taraxacum platycarpum H. Dahlst. showed very weak cytotoxic effects on Vero cells at > $100\;{\mu}g/ml$ but acycloguanosine showed strang cytotoxic effects on Vera cells at > $100\;{\mu}g/ml$. As a result, #3, #73, MW-69 and CW-18 are considered as potentially useful for anti-HSV-1 agent and will be the focus of further research. Abbreviations: CM - methanol extracts of traditional prescriptions; CW - water extracts of traditional prescriptions; MM - methanol extracts of individual herbs; MW - water extracts of individual herbs.

• Journal of Bio-Environment Control
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• v.26 no.3
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• pp.181-187
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• 2017

The purpose of this study was to provide basic data for development of environmental design technology for greenhouses constructed in reclaimed lands. The climatic conditions around seven major reclaimed land areas with a plan to install advanced horticultural complexes in Korea were analyzed. The characteristics of natural ventilation and temperature rise through the thermal environment measurement of the greenhouse in Saemangeum were analyzed. The part to be applied to the environmental design of the greenhouses in reclaimed lands were reviewed. Results of comparing the ventilation rate of the greenhouse according to the presence or absence of plants showed the greenhouse with plants had the lower ventilation rate, but the smaller rise of indoor temperature due to the evapotranspiration of plants. In the greenhouse with plants, the number of air changes was in the range of 0.3 to 0.9 volumes/min and the average was 0.7 volumes/min. The rise of indoor temperature relative to outdoor temperature was in the range of 1 to $5^{\circ}C$ and the average $2.5^{\circ}C$. The natural ventilation performance of the experimental greenhouse constructed in the reclaimed land almost satisfied the recommended ventilation rate in summer and the rise of indoor temperature relative to outdoor temperature did not deviate considerably from the cultivation environment of plants. Therefore, it was determined that the greenhouse cultivation in Saemangeum reclaimed land is possible with only natural ventilation systems without cooling facilities. As the reclaimed land is located in the seaside, the wind is stronger than the inland area, and the fog is frequent. This strong wind speed increases the ventilation rate of greenhouses, which is considered to be a factor for reducing the cooling load. In addition, since the fog duration is remarkably longer than that of inland area, the seasonal cooling load is expected to decrease, which is considered to be advantageous in terms of the operation cost of cooling facilities.