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Effect of TGF-${\beta}$ Supplementation on In Vitro Maturation of Hanwoo COCs (Cumulus Oocytes Complexes) (TGF- ${\beta}$ 첨가가 한우 난포란의 체외성숙에 미치는 영향)

  • Choi, Sun-Ho;Lee, Hye-Hyun;Yeon, Seong-Heum;Han, Man-Hye;Kim, Hyun-Jong;Cho, Sang-Rae;Woo, Jae-Seok;Baek, Kwang-Soo;Ryu, Il-Sun;Son, Dong-Soo
    • Development and Reproduction
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    • v.8 no.2
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    • pp.119-122
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    • 2004
  • It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs in vitro. Attemps had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of TGF-${\beta}$ on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in TCM 199 containing 0.1, 1 or 10 ng/ml TGF-${\beta}$ for 24 hrs, metaphaseⅡ of COCs were obtained 95.8%, 100% of matured COCs, respectively and there were no differences among the concentrations of TGF-${\beta}$. Matured COCs with TGF-${\beta}$ cultured in maturation medium after in vitro fertilization, developmental rate to blastocyst were 0~0.8%. Matured COCs with TGF-${\beta}$ were cultured in TCM 199+10% FBS, 0.8% BSA, 0.1% PVA, blastocyst formation were showed in 12.4%, 12.8%, 8.5% of those and cultured in IVMD or IVD without serum were 38.4%, 34.8%, respectively. There were significant differences among the media (P<0.05). TGF-${\beta}$ is available for i vitro maturation of bovine COCs, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine development.

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Electrofusion of Yeast Cells and Their Genetic Analysis Using RAPD-PCR (효모세포의 전기융합 및 융합세포의 RAPD-PCR을 이용한 유전적 분석)

  • Kim, Seung;Kim, Jae-Sung;Sapkota, Kumar;Park, In-Sung;Cho, Moon-Gu;Park, Yeal;Chun, Hong-Sung;Choi, Bong-Suk;Park, Se-Eun;Choi, Han-Suk;Kim, Myung-Kon;Kim, Sung-Jun
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.186-191
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    • 2006
  • In the present study, an attempt has been made to produce hybrid yeast strains of different useful and dominant characteristics. The hybrid yeast strains were produced by electrofusion and their genetic analysis were performed by RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction). The protoplast of Saccharomyces cerevisiae KCTC 7904 and Zygosaccharomyces rouxii KCTC 7966 were obtained above 92% when treated with lyticase at $30^{\circ}C$ for $60{\sim}90$ min after the pretreatment of $1{\sim}2%$ 2-mercaptoethanol at $30^{\circ}C$ for $15{\sim}20$ min. The fusant was produced from paired protoplast stage under the electric pulse at high frequency conditions (1.5 MHz/50 pV, 615 $V/256\;{\mu}sec$) within glass-platinum made electrofusion chamber. Changes in RAPD patterns in mother cells and hybrid cells proved that the fusant contains two types of yeast gene originated from its parent. Furthermore, fermentation characters exhibits by the fusant cell confirmed its genetic changes. These results suggest that genetically stable hybrid yeast strains of economic importance can be produced by electrofusion technique and these electrofused yeast cells have an enormous impact in biotechnology and biomedicine.

Analysis and countermeasure of causes of inducing violence of private security companies on the actual sites of administrative execution by proxy (행정대집행 현장에서 민간경비업체의 폭력 유발 원인 분석과 대책)

  • Choi, Kee-Nam
    • Korean Security Journal
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    • no.18
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    • pp.119-141
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    • 2009
  • Administrative execution by proxy is one of forced executions of administration and is also called as "enforced execution by proxy" in which administration institutions or the third party executes by proxy on behalf of parties who did not execute obligations under administration law and files claims to compensate expenses required in the proxy execution. Despite the actual site of administrative execution by law, social problems are generated because various violence and behaviors of infringement of human rights between executer and obligator are rampant and thus causing human damages since forced execution by physical force is carried out and cases of police indictments and petition to human rights committee are gradually increasing. Majority of people mobilized in this actual site of violence are supplied by private security companies which provide service contract and mobilization of people without qualification of guards or security service and irrational execution by proxy and violent actions by so-called service hooligans connected to violence organizations are now becoming social issues. In these actual sites of violence, structurally very complicated problems such as economic rights, right of residence, struggle for living, and intervention by outsiders are contained. This thesis has analyzed causes of outbreaks of violence and discussed about improvement countermeasure by paying attention to mobilization of people by private security companies. As the result, through revision and improvement of laws and systems, execution institution and policemen must be present at actual sites of execution by proxy to control physical execution of private security companies to be carried out legally and when violent collisions are occurring, it shall be stipulated that police should immediately intervene. Practices of execution by proxy of execution administration institutions shall be avoided and causes of occurrences of violence shall be eliminated by discrete decisions of execution by proxy, elimination of service contract conditions focused on accomplishments, and stipulation of responsibility of execution institutions when problems occur. Practices of solving petitions through collective actions of obligators shall be eliminated and strict enforcement of laws such as disturbance of official execution or compensation claims for expenses of execution by proxy must be carried out and intervention by the third parties must be intercepted. Mobilization of manpower by security companies shall be limited to people with prior registration who have acquired and finished qualification and education by security business law and before putting them on actual sites, it shall be obliged that execution plan with clear written records of working location, mission, and work rules must be submitted in advance to police station in charge and also they must be controlled to follow laws and statutes such as uniform and equipments. In addition, personal criminal responsibility for violent actions must be clearly stipulated and advanced securing soundness of security companies such as limits of service contracts with records of accidents is required. Order placement behaviors of special organizations under the pretext of rehabilitation business must be eradicated and companies with capability and strong intention of observation of laws must be able to receive orders by intercepting chains of contracts and sub-contracts. Issues of improvement countermeasure of social problem, living, and compensation including rights of residence and environment are excluded from the discussion.

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The Comparison and Index Components in Quality of Salt-Fermented Anchovy Sauces (멸치 액젓의 품질 비교 및 품질 지표성분에 관한 연구)

  • Oh, Kwang-Soo
    • Korean Journal of Food Science and Technology
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    • v.27 no.4
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    • pp.487-494
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    • 1995
  • To assay the quality of anchovy sauce, 10 kinds of commercial anchovy sauce(CAS) were purchased from markets and traditional anchovy sauce(TAS) were prepared. And their physicochemical-microbial characteristics were compared. The compositions of CAS were as followed; pH $5.5{\sim}5.7$, salinity $21.0{\sim}23.2%$, VBN $92.8{\sim}305.4\;mg/100g$, total nitrogen $928.0{\sim}1870.9\;mg%$, amino-nitrogen $338.6{\sim}680.3\;mg%$, and acidity $11.58{\sim}24.58\;ml$. The CAS was lower in pH, smaller in contents of VBN, total-N, amino-N and larger in contents of moisture, salinity than TAS. In Hunter values, CAS was generally lower in L, b values whereas higher in a and ${\Delta}E$ values than TAS. Viable cell counts on 0% NaCl-medium of CAS and TAS were $6.4{\times}10^1{\sim}3.0{\times}10^5\;and\;8.7{\times}10^4$, and those on 2.5% NaCl-medium were $0.8{\times}10^2{\sim}2.2{\times}10^5\;and\;1.6{\times}10^4{\sim}4.5{\times}10^5$, respectively. These viable cell counts in CAS and TAS were gradually decreased according to storage time. In composition of extractives, total free amino acid contents of CAS and TAS were $5498.5{\sim}12123.8\;mg%$, 12797.9 mg%, and these contents were gradually decreased during storage. The major amino acids were found glutamic acid, alanine and leucine in CAS, and alanine, glutamic acid, leucine and valine in TAS. Also contents of hypoxanthine, TMAO, TMA in CAS and TAS were shown $86.4{\sim}161.2\;mg%,\;51.6{\sim}99.2\;mg%,\;23.2{\sim}42.9\;mg%$ and 103.7 mg%, 128.8 mg%, 55.8 mg%, respectively. We may conclude from the results of present experiments that parts of tested CAS were somewhat putrefied and there was a great difference in the quality compared with TAS, whereas TAS maintained good conditions for preserving the quality until storage 2 years.

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Isolation and Characteristics of a Phenol-degrading Bacterium, Rhodococcus pyridinovorans P21 (페놀분해세균 Rhodococcus pyridinovorans P21의 분리 및 페놀분해 특성)

  • Cho, Kwang-Sik;Lee, Sang-Mee;Shin, Myung-Jae;Park, Soo-Yun;Lee, Ye-Ram;Jang, Eun-Young;Son, Hong-Joo
    • Journal of Life Science
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    • v.24 no.9
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    • pp.988-994
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    • 2014
  • The effluents of chemical and petroleum industries often contain non-biodegradable aromatic compounds, with phenol being one of the major organic pollutants present among a wide variety of highly toxic organic chemicals. Phenol is toxic upon ingestion, contact, or inhalation, and it is lethal to fish even at concentrations as low as 0.005 ppm. Phenol biodegradation has been studied in detail using bacterial strains. However, these microorganisms suffer from substrate inhibition at high concentrations of phenol, whereby growth is inhibited. A phenol-degrading bacterium, P21, was isolated from oil-contaminated soil. The phenotypic characteristics and a phylogenetic analysis indicated the close relationship of strain P21 to Rhodococcus pyridinovorans. Phenol biodegradation by strain P21 was studied under shaking condition. The optimal conditions for phenol biodegradation by strain P21 were 0.09% $KNO_3$, 0.1% $K_2HPO_4$, 0.3% $NaH_2PO_4$, 0.015% $MgSO_4{\cdot}7H_2O$, 0.001% $FeSO_4{\cdot}7H_2O$, initial pH 9, and $20-30^{\circ}C$, respectively. When 1,000 ppm of phenol was added to the optimal medium, the strain P21 completely degraded it within two days. Rhodococcus pyridinovorans P21 could grow in up to 1,500 ppm of phenol as the sole carbon source in a batch culture, but it could not grow in a medium containing above 2,000 ppm. Moreover, strain P21 could utilize toxic compounds, such as toluene, xylene, and hexane, as a sole carbon source. However, no growth was detected on chloroform.

tans-Resveratrol Content of Varieties and Growth Period in Peanut (땅콩 품종 및 생육기별 trans-Resveratrol 함량)

  • Lee, Mi-Ja;Cheong, Young-Keun;Kim, Hyung-Soon;Park, Ki-Hun;Doo, Hong-Soo;Suh, Duck-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.6
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    • pp.429-433
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    • 2003
  • trans-Resveratrol(3,5,4'-trihydroxystilbene) is phenolic compound present in grapes, wines, and peanuts, has been reported to have health benefits including anticarcinogenic effects, protection against cardiovascular diseases and reduced cancer risk. A simple method for the quantitative extraction of trans-resveratrol from peanut has been developed. Optimal conditions for extraction were investigated. Type of solvent, time, and temperature assayed influenced trans-resveratrol yield. Adequate extraction condition was decided to ethanol/water (80:20v/v) maintained at $25^{\circ}C$ for 45 min. After extraction, the protocol consists of sample preparation using a $\textrm{C}_{18}$ solid-phase extraction (SPE) cartridge after concentrate with rotary evaporator and quantified by reversed phase HPLC using a $\textrm{C}_{18}$ column at 308 nm. Analytical methods for measuring trans-resveratrol in peanut were adapted to isolate, identify, and quantify trans-resveratrol in 11 peanut varieties by HPLC (high-performance liquid chromatography) with UV detector, The 11 peanut varieties content ranged from 0.018 to 1.125 $\mu\textrm{g}/\textrm{g}$ with an average of 0.289 $\mu\textrm{g}/\textrm{g}$. The contents were higher in the seeds with than without testa, regardless of varieties. The trans-resveratrol content was Higher in 110, 130 days after sowing than that of other period.

Micropropagation of Oak Seedlings from 37 Plus Half-Sib Families (참나무류(類) 수형목(秀型木) 37가계(家系)의 기내증식(器內增殖))

  • Moon, Heung Kyu;Youn, Yang;Son, Sung Ho;Lee, Suk Koo;Yi, Jae Sun
    • Journal of Korean Society of Forest Science
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    • v.82 no.1
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    • pp.26-33
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    • 1993
  • In vitro shoot proliferation and rooting were tested for 2-0 seedlings of half-sib families of 4 plus oaks trees. Nodal segments having axillary buds from 37 families(16 of Quercus acutissima, 10 of Q. variabilis, 7 of Q. serrata, and 4 of Q. mongolica) were cultured on WPM(Woody Plant Medium) supplemented with 0.5 mg/l BA (6-benzyladenine) and 0.01 mg/l NAA(${\alpha}$-naphthalene acetic acid) and subcultured at 2-3 weeks of intervals fur 6 months. In vitro rooting was carried out on GD(Gresshoff and Doy) medium supplemented with 0.5mg/l IBA(indole butyric acid). The capacity for shoot proliferation and rooting was highly varied with families. Generally, white oaks(Q. serrata and Q. mongolica) showed poor response than black oaks(Q. acutissima and Q, variabilis) in shoot proliferation and rooting. Among the total of 37 families, 7 of Q. acutissima, each 2 of Q. variabilis, Q. serrata, and Q. mongolica revealed abilities for continuous shoot proliferation, and the others failed to proliferate. Rooting of the selected oak trees also greatly varied among the families. In Q. acutissima, rooting ratio ranged from 10.0%(CB 25. KG 4) to 89.8%(CB 18). Although 26.7% of KG 16 in Q. variabilis, 3.3% of JN 15 in Q. serrata were rooted, Q. mongolica was not rooted at all in this experimental conditions. No relationship between shoot growth and the rooting ability was observed. Present results suggest the possibility of large-scale micropropagation, but further studies on family differences, shoot-tip necrosis, and callusing of rooting junction are still required to develop reliable micropropagation systems.

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Expression and Purification of the Phosphatase-like Domain of a Voltage-Sensing Phosphatase, Ci-VSP (막 전위 감지 탈인산화 효소, Ci-VSP의 유사 탈인산화 효소 도메인의 발현과 정제)

  • Kim, Sung-Jae;Kim, Hae-Min;Choi, Hoon;Kim, Young-Jun
    • 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.

Influences of Roasted or Non-roasted Brown Rice Addition on the Nutritional and Sensory Properties and Oxidative Stability of Sunsik, Korean Heated Cereal Powder (현미의 볶음 처리 유무와 첨가량이 선식의 영양적 관능적 특성 및 산화안정성에 미치는 영향)

  • Lee, Byung-Yong;O, Jin-Hwan;Kim, Mi-Hyun;Jang, Ki-Hyo;Lee, Jae-Cheol;Surh, Jeong-Hee
    • Korean journal of food and cookery science
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    • v.26 no.6
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    • pp.872-886
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    • 2010
  • Six kinds of sunsik containing different contents of brown rice(BR; 20, 30, and 50%) were prepared and subjected to various processing conditions(with or without roasting at $200^{\circ}C$ for 20 min e.g., designated as RBR50 or BR50) to assess their functionality as ready-to-eat foods. They were also assessed for their nutritional and sensory properties and oxidative stability. Dietary fiber contents were proportionate to the levels of added BR. Protein was highest in RBR50 (p<0.001), which also had the highest amounts of free and structural amino acids. The amount of free amino acids tended to increase with roasting, although most amino acids were present in structural form. Oleic acid and linoleic acid were the predominant fatty acids in all prepared sunsik, and RBR50 presented noticeably higher peroxidability index due to its higher amount of linoleic acid(p<0.05). Nevertheless, RBR50 showed good oxidative stability; this phenomenon was observed in all sunsik with roasted BR but not in those with non-roasted BR. It is implied that potential antioxidants might have been newly formed or converted from their precursors while BR was roasted. Roasting process also had an impact on the sensory properties of sunsik, e.g., sunsik with added roasted BR showed lower dissolution and darker color intensity compared to its counterpart sunsik.

Study on Effective Preservation of Bovine Pericardium Using Decellulariation and ${\alpha}$-galactosidase for Eliminating Xenoreactive Antigen (이종 항원 제거를 위한 무세포화와 알파-갈락토시다아제를 이용한 효과적인 우심낭 보존 방법에 관한 연구)

  • Kim, Min-Seok;Park, Cham-Jin;Kim, Soo-Hwan;Lim, Hong-Gook;Kim, Yong-Jin
    • Journal of Chest Surgery
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    • v.43 no.6
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    • pp.576-587
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    • 2010
  • Background: Effective decellularization and fixation process is critical, in order to use xenogenic valves clinically. In the present study, we decellularized bovine pericardium using sodium dodecyl sulfate (SDS) and N-lauroyl sarcosinate, treated with $\alpha$-galactosidase, and then fixed in various manners, to find out the most effective tissue preservation & fixation procedure. Material and Method: Bovine pericardium was decellularized with SDS and N-lauroyl sarcosinate, and treated with $\alpha$-galactosidase. Both groups were fixed differently, by varying glutaraldehyde (GA) or EDC (1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide)/N-hydroxysuccinamide (NHS) treatment conditions. Thereafter, physical examination, tensile strength test, thermal stability test, cytotoxicity test, pronase test, pronase-ninhydrin test, purpald test, permeability test, compliance test, H&E staining, DNA quantification, and $\alpha$-galactose staining were carried out to each groups. Result: GA fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, EDC/NHS-GA dual fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, and showed better thermal stability than GA fixed groups. In pronase test and pronase-ninhydrin test, GA fixed groups and EDC/NHS-GA dual fixed groups showed stronger crosslinks than EDC/NHS groups. Permeability and compliance tended to increase in EDC/NHS-GA dual fixed groups, compared to GA fixed groups. But, EDC/NHS-GA dual fixed groups had stronger tensile strength and lower cytotoxicity than GA fixed groups. Conclusion: We have verified that EDC/NHS-GA dual fixation can make effective crosslinks and lower the toxicity of GA fixation. Henceforth, we will verify if EDC/NHS-GA dual fixation can lower calcifications & tissue failure in vivo experiment.