• Journal of Embryo Transfer
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• v.33 no.4
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• pp.245-254
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• 2018

Pigs are considered as optimal donor animal for the successful xenotransplantation. To increase the possibility of clinical application, genetic modification to increase compatibility with human is an important and essential process. Genetic modification technique has been developed and improved to produce genetically modified pigs rapidly. CRISPR/Cas9 system is widely used in various fields including the production of transgenic animals and also can be enable multiple gene modifications. In this study, we developed new gene targeting vector and enrichment system for the rapid and efficient selection of genetically modified cells. We conducted co-transfection with two targeting vectors for simultaneous inactivation of two genes and enrichment of the genetically modified cells using MACS. After this efficient enrichment, genotypic analysis of each colony showed that colonies which have genetic modifications on both genes were confirmed with high efficiency. Somatic cell nuclear transfer was conducted with established donor cells and genetically modified pigs were successfully produced. Genotypic and phenotypic analysis of generated pigs showed identical genotypes with donor cells and no surface expression of ${\alpha}$-Gal and HD antigens. Furthermore, functional analysis using pooled human serum revealed dramatically reduction of human natural antibody (IgG and IgM) binding level and natural antibody-mediated cytotoxicity. In conclusion, the constructed vector and enrichment system using MACS used in this study is efficient and useful to generate genetically modified donor cells with multiple genetic alterations and lead to an efficient production of genetically modified pigs.

• Korean Journal of Food Science and Technology
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• v.19 no.3
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• pp.225-230
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• 1987

The foam separation of bovine serum proteins was investigated and the protein fractionation by foam separation was analyzed by PAG electrophoresis. The protein concentration for the surface excess formation of bovine serum was in the range of $20-800\;{\mu}g/ml$. At pH 5, the foamate volume was maximum, but the enrichment ratio minimum. As the temperature was elevated, the foamate volume decreased and the enrichment ratio increase. As the gas flow rate increased from 25 to 100 ml/min, the foamate volume decreased and the enrichment ratio increased. The enrichment ration became maximum when the added ionic strength of serum solution was in the range of 1-3 by the addition of different types of salts, and this was related to the reduction of surface tension of the solution. In general, BSA, ${\alpha}_1$, and ${\alpha}_2-globulins$, which have relatively small molecular weight and high hydrophobicity, moved easily to the foam, and the separation of protein fractions in the serum varied with the changes in pH, temperature, gas flow rate and ionic strength of the solution.

• Journal of Sensor Science and Technology
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• v.27 no.4
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• pp.237-241
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• 2018

The real-time enrichment and detection of pathogens are serious issues and rapidly evolving field of research because of the ability of these pathogens to cause infectious diseases. In general, bacterial detection is accomplished by conventional colony counting or by polymerase chain reaction (PCR) after DNA extraction. As colony counting requires considerable time to cultivate, PCR is an attractive method for rapid detection. A small number of pathogens can cause diseases. Hence, a pretreatment process, such as enrichment is essential for detecting bacteria in an actual environment. Thus, in this study, we developed a microfluidic chip capable of performing rapid enrichment of bacteria and the extraction of their genes. A lectin, i.e., Concanavalin A (ConA), which shows binding affinity to the surface of most bacteria, was coated on the surface of magnetic particles to nonspecifically capture bacteria. It was subsequently concentrated through magnetic forces in a microfluidic channel. To lyse the captured bacteria, magnetic particles were irradiated by a wavelength of 532nm. The photo-thermal effect on the particles was sufficient for extracting DNA, which was consequently utilized for the identification of bacteria. Our device will help monitor the existence of bacteria in various environmental situations such as water, air, and soil.

• Journal of Microbiology and Biotechnology
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• v.17 no.1
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• pp.110-115
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• 2007

Electrochemically active bacteria were successfully enriched in an electrochemical cell using a positively poised working electrode. The positively poised working electrode (+0.7 V vs. Ag/AgCl) was used as an electron acceptor for enrichment and growth of electrochemically active bacteria. When activated sludge and synthetic wastewater were fed to the electrochemical cell, a gradual increase in amperometric current was observed. After a period of time in which the amperometric current was stabilized (generally 8 days), linear correlations between the amperometric signals from the electrochemical cell and added BOD (biochemical oxygen demand) concentrations were established. Cyclic voltammetry of the enriched electrode also showed prominent electrochemical activity. When the enriched electrodes were examined with electron microscopy and confocal scanning laser microscopy, a biofilm on the enriched electrode surface and bacterium-like particles were observed. These experimental results indicate that the electrochemical system in this study is a useful tool for the enrichment of an electrochemically active bacterial consortium and could be used as a novel microbial biosensor.

• Journal of Life Science
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• v.27 no.4
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• pp.435-441
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• 2017

Perchlorate ($ClO_4^-$) is an emerging contaminant detected in soil, groundwater, and surface water. Previous study revealed bacterial community in the enrichment culture tdegraded perchlorate using elemental sulfur as an electron donor. Quantitative and qualitative molecular methods were employed in this study to investigate archaeal community in the enrichment culture. Real-time qPCR showed that archaeal 16S rRNA gene copy number in the culture was about 1.5% of bacterial 16S rRNA gene copy number. This suggested that less archaea were adapted to the environment of the enrichment culture and bacteria were dominant. DGGE banding pattern revealed that archaeal community profile of the enrichment culture was different from that of the activated sludge used as an inoculum for the enrichment culture. The most dominant DGGE band of the enrichment culture was affiliated with Methanococci. Further research is necessary to investigate metabolic role of the dominant archaeal population to better understand microbial community in the perchlorate-reducing enrichment culture.

• Journal of Food Hygiene and Safety
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• v.38 no.6
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• pp.449-456
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• 2023

In this study, we developed Rapid Enrichment Broth for Vibrio parahaemolyticus (REB-V), a broth capable enriching V. parahaemolyticus from 10⁰ CFU/mL to 10⁶ CFU/mL within 6 hours, which greatly facilitates the rapid detection of V. parahaemolyticus. Using a modified Gompertz model and response surface methodology, we optimized supplement sources to rapidly enrich V. parahaemolyticus. The addition of 0.003 g/10 mL of D-(+)-mannose, 0.002 g/10 mL of L-valine, and 0.002 g/10 mL of magnesium sulfate to 2% (w/v) NaCl BPW was the most effective combination of V. parahaemolyticus enrichment. Optimal V. parahaemolyticus culture conditions using REB-V were at pH 7.84 and 37℃. To confirm REB-V culture efficiency compared to 2% (w/v) NaCl BPW, we assessed the amount of enrichment achieved in 7 hours in each medium and extracted DNA samples from each culture every hour. Real-time PCR was performed using the extracted DNA to verify the applicability of this REB-V culture method to molecular diagnosis. V. parahaemolyticus was enriched to 5.452±0.151 Log CFU/mL in 2% (w/v) NaCl BPW in 7 hours, while in REB-V, it reached 7.831±0.323 Log CFU/mL. This confirmed that REB-V enriched V. parahaemolyticus to more than 10⁶ CFU/mL within 6 hours. The enrichment rate of REB-V was faster than that of 2% (w/v) NaCl BPW, and the amount of enrichment within the same time was greater than that of 2% (w/v) NaCl BPW, indicating that REB-V exhibits excellent enrichment efficiency.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.2
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• pp.106-117
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• 2006

We measured the concentrations of organic matter and metallic elements (Al, Fe, Cr, Mn, Ni, Cu, Zn, As, Cd, Pb and Hg) in the surface sediments of Masan Harbor (in the southern sea, Korea) to evaluate the geochemical characters of sediment and the pollutions by organic matter and metallic elements. The mean grain size of the surface sediments in the study area ranged from $5.6{\phi}$ to $7.8{\phi}$, indicating silt sediment. The water content of the surface sediments exceeded 60% except at some stations. The contents of ignition loss (IL), total organic carbon (TOC) and total nitrogen (TN) ranged from 7.2-14.3%, 1.2-3.2%, and 0.10-0.28%, respectively. Based on the C/N ratios, the organic matter in the surface sediments of Masan Harbor may originate from terrigenous sources including fluvial inputs (mainly sewage in urban areas). The chemical oxygen demand (COD) and acid volatile sulfide (AVS) ranged from $11.3-29.9\;mgO_2/g\;dry$ and 0.20-4.47 mgS/g dry, respectively, and low concentrations were observed near a shipping route. In addition, the concentrations of metallic elements showed large spatial variations in Masan Harbor and the distributions of metallic elements were also comparable to those of organic matter. This implies that the distributions of organic matter and metallic elements in the surface sediments of Masan Harbor are mainly controlled by biogenic matter and artificial action (mainly dredging). In addition, we calculated the enrichment facto. (EF) and geoaccumulation index (Igeo) in order to evaluate pollution by metallic elements. The enrichment of metallic elements relative to Al was three to eighteen times higher at the study sites, compared to levels in the Earth's crust except for Fe, Ni and Mn. In addition, the Igeo class indicated that the surface sediments in the study area were moderately to strongly polluted in terms of metallic elements.

• Proceedings of KOSOMES biannual meeting
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• 2007.05a
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• pp.27-33
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• 2007

Heavy metals in the surface seawaters and sediments were measured in Jinhae Bay. The high concentrations of heavy metals in the seawaters were found at the stations near the islands. In the sea waters, the mean concentrations of dissolved heavy metals except for Pb were not higher than previous data in this bay. Higher heavy metal contents in the surface sediments were observed at the stations adjacent to the Kojedo or Kadukdo of the Kadoksudo. The contents of Co, Ni, Zn, Cu, As and Cd in the surface sediments showed relatively high correlation coefficients with IL and COD. The order of enrichment factors(EFs) of heavy metals in the sediments was As>Cd>Pb>Zn>Co>Cu>Hg>Ni, and the EFs of As, Cd, Pb and Zn were higher than 1.

• Corrosion Science and Technology
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• v.21 no.6
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• pp.466-475
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• 2022

Surface oxides and intergranular (IG) oxidation phenomena in Alloy 600 depending on hydrogen concentration were characterized to obtain clear insight into the primary water stress corrosion cracking (PWSCC) behavior upon exposure to pressurized water reactor primary water. When hydrogen concentration was between 5 and 30 cm³ H₂/kg H₂O, NiFe₂O₄ and NiO type oxides were found on the surface. NiO type oxides were found inside the oxidized grain boundary when hydrogen concentration was 5 cm³ H₂/kg H₂O. However, only NiFe₂O₄ spinel on the surface and Ni enrichment were observed when hydrogen concentration was 30 cm³ H₂/kg H₂O. These results indicate that the oxidation/reduction reaction of Ni in Alloy 600 depending on hydrogen concentration can considerably affect surface oxidation behavior. It appears that the formation of NiO type oxides in a Ni oxidation state and Ni enrichment in a Ni reduction (or metallic) state are common in primary water. It is believed that the above different oxidation/reduction reactions of Ni in Alloy 600 depending on hydrogen concentration can also significantly affect the resistance to PWSCC of Alloy 600.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.6
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• pp.777-787
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• 2017

Organic enrichment and pollution was investigated in surface sediments from Jinhae Bay and Geoje-Hansan Bay of Korea, which contain the largest oyster farms in Korean coastal areas. Geochemical indicators (chemical oxygen demand, total organic carbon, ignition loss, and acid volatile sulfide) in sediments, ammonium and nitrate in pore water, and bioluminescence inhibition test for sediment extracts were analyzed. Temporal changes of organic enrichment were also investigated using sediment core samples from Geoje-Hansan Bay. The level of organic pollution in sediments from Jinhae Bay was significantly greater than that of sediments from Geoje-Hansan Bay. Compared with other sites, Jinhae Bay was one of the most polluted coastal areas of Korea. The levels of geochemical indicators in May were comparable to, or higher than, in August. Ammonium concentrations in pore water were two orders of magnitudes greater than the nitrate concentrations, suggesting that the bays are reducing environments. The concentrations of total organic carbon in core sediment samples from shellfish-farming areas increased significantly from 2000 to the present year, and it seems to be associated with increases in anthropogenic activities.