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Purification and Characterization of 5,10-Methenyltetrahydrofolate Synthetase from Chicken Liver (닭의 간 유래의 5,10-Methenyltetrahydrofolate Synthetase의 정제 및 특성)

  • Cho, Yong-Kweon
    • Journal of the Korean Chemical Society
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    • v.54 no.5
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    • pp.567-572
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    • 2010
  • 5,10-Methenyltetrahydrofolate synthetase from chicken liver was purified through 30-70% ammonium sulfate fractionation, Q Sepharose Fast Flow anion exchange and Source 15Phe hydrophobic interaction chromatography. Specific activities of cell extract, ammonium sulfate, Q Sepharose Fast Flow and Source 15Phe were 0.0085, 0.031, 0.80 and 1.27 U/mg, respectively. Purification fold activities of cell extract, ammonium sulfate, Q Sepharose Fast Flow and Source 15Phe were 1, 3.7, 94.1 and 149.4, respectively. HPLC gel permeation chromatography and SDS-polyacrylamide electrophoresis experiments indicated that the enzyme is a monomeric protein with a molecular weight of 22.8 kDa. Km for 5-methyl THF and Mg-ATP were $7.1\;{\mu}M$ and $63\;{\mu}M$, respectively. Optimum temperature and pH were $30^{\circ}C$ and 6.0, respectively. The data for metal ion specificity and stoichiometry showed that the maximum activity was obtained with a 1:l. ratio of $Mg^{2+}$. The ATP and Km values increased in the order of MgATP, MgCTP, MgUTP and MgGTP, and the maximum activities also decreased in the same order, indicating MgATP as the most efficient substrate. The enzyme was chemically modified only by tetranitrometane and 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide, indicating that tyrosine and carboxylate are present in the active site.

Biochemical Characterization of the Dual Positional Specific Maize Lipoxygenase and the Dependence of Lagging and Initial Burst Phenomenon on pH, Substrate, and Detergent during Pre-steady State Kinetics

  • Cho, Kyoung-Won;Jang, Sung-Kuk;Huon, Thavrak;Park, Sang-Wook;Han, Ok-Soo
    • BMB Reports
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    • v.40 no.1
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    • pp.100-106
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    • 2007
  • The wound-inducible lipoxygenase obtained from maize is one of the nontraditional lipoxygenases that possess dual positional specificity. In this paper, we provide our results on the determination and comparison of the kinetic constants of the maize lipoxygenase, with or without detergents in the steady state, and characterization of the dependence of the kinetic lag phase or initial burst, on pH, substrate, and detergent in the pre-steady state of the lipoxygenase reaction. The oxidation of linoleic acid showed a typical lag phase in the pre-steady state of the lipoxygenase reaction at pH 7.5 in the presence of 0.25% Tween-20 detergent. The reciprocal correlation between the induction period and the enzyme level indicated that this lag phenomenon was attributable to the slow oxidative activation of Fe (II) to Fe (III) at the active site of the enzyme as observed in other lipoxygenase reactions. Contrary to the lagging phenomenon observed at pH 7.5 in the presence of Tween-20, a unique initial burst was observed at pH 6.2 in the absence of detergents. To our knowledge, the initial burst in the oxidation of linoleic acid at pH 6.2 is the first observation in the lipoxygenase reaction. Kinetic constants (Km and kcat values) were largely dependent on the presence of detergent. An inverse correlation of the initial burst period with enzyme levels and interpretations on kinetic constants suggested that the observed initial burst in the oxidation of linoleic acid could be due to the availability of free fatty acids as substrates for binding with the lipoxygenase enzyme.

An Experimental Study for Performance Evaluation in Dogs of Preventive Contrast Media Extravasation with a Strain Gage Based Prototype Extravasation Detection Accessory System (잡견에서 조영제 혈관외유출 예방을 위한 스트레인 게이지 기반의 EDA 시스템 성능 평가를 위한 실험적 연구)

  • Kweon, D.C.;Yoo, B.G.;Lee, J.S.;Cho, M.S.;Yang, S.H.
    • Journal of Biomedical Engineering Research
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    • v.29 no.1
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    • pp.66-72
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    • 2008
  • The major risk associated with the use of automated power injectors is the well known complication of contrast material extravasation at the injection site. Automated injection of computed tomography (CT) contrast media can produce the compartment syndrome. The purpose of this study was to assess the ability of this device during clinically important episodes of extravasation. The extravasation detection accessory (EDA) system was composed of a strain gage, an amplifier and a computer based system. A strain gage pliable adhesive patch was applied to the skin aver the intravenous catheter and the catheter was connected to the power injector with a cable to monitor the resolution data. If the programmed monitoring, which was developed with MS Visual C++, at the extravasation occurred, then the injection was interrupted the auto injector. CT was used to demonstrate the clinically important extravasation. This study was a prospective, observational study in which the EDA system was used to monitor the automated mechanical injection of contrast material in 7 dogs. There were two true-positive cases (range of extravasation volumes: $18{\sim}22ml$), twenty three true-negative cases, three false-positive cases and no false-negative cases. The EDA system had a sensitivity of 100% and a specificity of 88% for the detection of clinically important extravasation. The EDA system had good sensitivity for the detection of clinically important extravasation and the EDA system has the clinical potential for the early detection of extravasation of the contrast medium that is administered with power injectors. The EDA system is easy to use safe and accurate for the monitoring extravasation of the intravenous injections, and this system may prove especially useful in CT applications.

THE EFFECT OF PDGF-BB AND IGF-I COMBINATION ON THE HEALING OF ARTIFICIAL PERIAPICAL LESIONS IN BEAGLE DOGS (PDGF-BB와 IGF-I 혼합 투여가 비글견 인공 치근단 병소의 치유에 미치는 영향에 관한 연구)

  • Kim, Mi-Ri;Kim, Min-Kyum;Yoon, Soo-Han
    • Restorative Dentistry and Endodontics
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    • v.25 no.1
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    • pp.1-16
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    • 2000
  • It is difficult to treat the endodontic apical perforation successfully. In this study, we hypothesized that the application of PDGF-BB and IGF-I into periapical perforation site may accelerate periapical healing and lead to bone deposition. And the specificity of osteonectin in periapical healing was investigated. The experiments were performed on the upper and lower 51 premolar teeth of 4 beagle dogs. The pulp chamber of each tooth was opened and the dental plaque was inserted into the canal for developing the periapical lesion for 5 weeks. Then, the roots were artificially perforated at the apex with the number 4 profile of .06 taper. In each step, standard periapical radiographs were taken to compare the size of lesion each other. The radiographs were scanned and analyzed by image analysis system. The mean and standard deviation of periradicular radiolucency ratios were calculated in each group. ANOVA was used for comparison. 51 premolars were grouped into 3 groups; control group, calcium hydroxide-treated group and calcium hydroxide plus growth factors-treated group. In the control group, the apical perforations were not sealed and obturated with gutta-percha and ZOE sealer by lateral condensation technique. In the experimental groups, the apical perforation were sealed with calcium hydroxide and with/without $4{\mu}g$ of PDGF-BB & IGF-I in cellulose gel and obturated by lateral condensation technique. Fluorescent bone markers were used to measure new bone formation. Following 2, 4, 12 weeks after experiment the dogs were sacrificed and histologic sections were prepared. Each tooth block including periapical lesion was sectioned mesiodistally. One half of the sections were decalcified with 6% nitric acid and processed by standard paraffin embedding technique. The sections were stained by hematoxylin and eosin, and immunostained for osteonectin. Histomorphometrical measurement of neoformed bone was performed using a light microscope. And the other half of the sections were prepared by undecalcified preparation, and confocal laser scanning microscopic investigations were done.

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Cloning and Expression of a Alkaline Protease from Bacillus clausii I-52 (Bacillus clausii I-52로부터 alkaline protease 유전자의 클로닝 및 발현)

  • Joo, Han-Seung;Choi, Jang Won
    • Journal of agriculture & life science
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    • v.45 no.6
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    • pp.201-212
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    • 2011
  • The alkaline protease gene was cloned from a halo-tolerant alkalophilic Bacillus clausii I-52 isolated from the heavily polluted tidal mud flat of West Sea in Inchon Korea, which produced a strong extracellular alkaline protease (BCAP). Based on the full genome sequence of Bacillus subtilis, PCR primers were designed to allow for the amplification and cloning of the intact pro-BCAP gene including promoter region. The full-length gene consists of 1,143 bp and encodes 381 amino acids, which includes 29 residues of a putative signal peptide and an additional 77-amino-acid propeptide at its N-terminus. The mature BCAP deduced from the nucleotide sequence consists of 275 amino acids with a N-terminal amino acid of Ala, and a relative molecular weight and pI value was 27698.7 Da and 6.3, respectively. The amino acid sequence shares the highest similarity (99%) to the nattokinase precursor from B. subtilis and subtilisin E precursor from B. subtilis BSn5. The substrate specificity indicated that the recombinant BCAP could hydrolyze efficiently the synthetic substrate, N-Suc-Ala-Ala-Pro-Phe-pNA,and did not hydrolyze the substrates with basic amino acids at the P1 site. The recombinant BCAP was strongly inhibited by typical serine protease inhibitor, PMSF, indicating that BCAP is a member of the serine proteases.

Investigation of the Molecular Diagnostic Market in Animals (동물 분자 진단 시장의 동향)

  • Park, Chang-Eun;Park, Sung-Ha
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.1
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    • pp.26-33
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    • 2019
  • Recently, the rapid growth of the companion animal market has led to the development of animal disease diagnosis kits. Therefore, the utility of the introduction of biomarkers for the development of animal molecular diagnostics is being reevaluated. A good biomarker should be precise and reliable, distinguish between normal and diseased states, and differentiate between different diseases. Recently reported genetic markers, tumor markers (cell free DNA, circulating tumor cells, granzyme, and skin tumors), and others (brucellosis, programmed death recovery-1, symmetric dimethylarginine, periostin, and cysteinyl leukotrien) have been developed. The biomarkers are used for risk prediction or for the screening, diagnosis, and monitoring of disease progression. The most important criteria for related biomarkers are disease specificity. Many potential biomarkers have emerged from laboratory and test studies, but they have not been validated in independent or large-scale clinical studies. Candidate biomarkers evaluate disease associations, verify the effectiveness of biomarkers for early detection and disease progression, and incorporate them into humans and animals. In the future, it will be necessary to reevaluate the utility of well-structured biomarker-based research and study the development of kits that can be used in on-site tests in accordance with the trends introduced in the diagnosis of animal diseases.

A Study on the Guidelines for the Development of Domestic Flight Test Operation Manual (국내 비행시험 운영 매뉴얼 개발 기준 연구)

  • Kim, Mu-Geun;Baek, Seung-Don;Song, Chan-Yong;Ahn, Dae-Whee;Han, Jeongho;Yoo, Beong-Seon;Kang, Ja-Young
    • Journal of Advanced Navigation Technology
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    • v.23 no.4
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    • pp.281-288
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    • 2019
  • A Flight test is an important process that must be performed to maintain airworthiness in the aircraft lifecycle, including aircraft development and certification. Flight test can be conducted for the purpose of demonstrating or verifying compliance with the applicable standard. Results of the flight test air accident analysis confirm that it is caused by a combination of various factors as well as aircraft. The International Civil Aviation Organization recommends the establishment and implementation of a safety management system to prevent aircraft accidents, but it does not reflect the specificity of the flight test. As such, advanced countries are providing separate criteria to assist stakeholder in conducting flight test safety management. This study investigates and analyzes relevant regulations of advanced countries, in accordance with the construction of a national flight performance test site in Korea, and proposes government-level guidelines to help stakeholders develop their flight test operation manuals. The guidelines discuss topics such as organizational composition, safety and risk management, flight operation, personnel management, and administrative issues.

Development and validation of a Korean Affective Voice Database (한국형 감정 음성 데이터베이스 구축을 위한 타당도 연구)

  • Kim, Yeji;Song, Hyesun;Jeon, Yesol;Oh, Yoorim;Lee, Youngmee
    • Phonetics and Speech Sciences
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    • v.14 no.3
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    • pp.77-86
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    • 2022
  • In this study, we reported the validation results of the Korean Affective Voice Database (KAV DB), an affective voice database available for scientific and clinical use, comprising a total of 113 validated affective voice stimuli. The KAV DB includes audio-recordings of two actors (one male and one female), each uttering 10 semantically neutral sentences with the intention to convey six different affective states (happiness, anger, fear, sadness, surprise, and neutral). The database was organized into three separate voice stimulus sets in order to validate the KAV DB. Participants rated the stimuli on six rating scales corresponding to the six targeted affective states by using a 100 horizontal visual analog scale. The KAV DB showed high internal consistency for voice stimuli (Cronbach's α=.847). The database had high sensitivity (mean=82.8%) and specificity (mean=83.8%). The KAV DB is expected to be useful for both academic research and clinical purposes in the field of communication disorders. The KAV DB is available for download at https://kav-db.notion.site/KAV-DB-75 39a36abe2e414ebf4a50d80436b41a.

A Study on the Method of Activation of Space of Gwangheemun Considering Historical and Cultural Speciality (역사·문화적 특수성을 고려한 광희문(光熙門)의 공간 활성화 방안 연구)

  • Kim, Ji Eun;Park, Eun Soo
    • Korea Science and Art Forum
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    • v.19
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    • pp.243-257
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    • 2015
  • The Cultural Heritage Administration has selected Seoul Fortress Wall as the representative heritage of Korea to be applied for being registered as UNESCO World Heritage and announced the plan to restore and organize it, which has increased the interest to the Seoul Fortress Wall, the Historical Site No. 10. The great work to make the heritage value of Walls, 4 Big Gates and 4 Small Gates composing the Seoul Fortress wall with the length of 18.627 km to be recognized worldwide has limits if it is focused only on the physical restoration. It is because the Seoul Fortress Wall represents the long historical and cultural value as the space of life which connects closely the capital city and its vicinity. We need the plan to discover and utilize historical and cultural contents of Seoul Fortress Wall and its vicinity. Especially, Gwangheemun, which is one of the four Small Gates of Seoul Fortress Wall, is a precious cultural heritage which represents the transition of fortification technology of Chosun period as the space representing ordinary people. However, now Gwangheemun and its vicinity does not stand out the charm because of passive accessibility, landscape falling behind and absence of program. This made the lack of domestic and overseas tourists and the convenient space and various contents. This reality is because the value of space has been considered simply as a cultural heritage without considering the traditional, historical and cultural specificity. Therefore, this study is aimed to find the meaning and value of Gwangheemun by discovering its own latent intangible cultural, historical and artistic resource, and to find the way to connect with Gwangheemun, the existing tangible traditional architectural space and the way of vitalizing Gwangheemun as a new space.

A Study for Selecting Protected Wildlife Species and Potential Habitats in Seoul Metropolitan City (서울시 보호야생동.식물 종 및 잠재 서식지 선정 연구)

  • Han, Bong-Ho;Lee, Kyong-Jae;Ki, Kyong-Seok;Choi, Byeong-Eon
    • Korean Journal of Environment and Ecology
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    • v.25 no.4
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    • pp.601-613
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    • 2011
  • The purpose of this study is to provide an protected wildlife species and potential habitats in Seoul through the overall present status of the species which live in Seoul. This study was performed in Seoul, the entire area of which is 605.52 $km^2$. We analyzed a total of 57 studies, including academic papers and research reports, for the natural ecology in Seoul. As for the results from the study, it was found that a total 1,907 species from 5 taxa were inhabiting and growing within the area. Among the total, plants of 1,656 species accounted for 86.7%, 197 species of wild birds made up 10.3%, 29 species of amphibians and reptiles consisted of 1.5%, and 27 species of mammalia totaled 1.4%. A total of 440 species except foreign species and species legally preserved by the Ministry of Environment, common species and uncertain species were selected as primary candidates. By taxon, it shows 212 species of plants, 180 species of wild birds, 24 species of mammalia, and 24 species of amphibians and reptiles. For extensive research subjects and research data limited in certain insect family, analysis by literature for discovered insects was excluded. The selection criteria for the second candidate species was based on extinction crisis, rarity, historicity and locality, academic value, habitat specificity, etc. The second candidate species from the primary species were selected through taxon experts' evaluations. As for the results of the selection of the second candidates, a total 77 species were selected through the experts' evaluations. Each species by taxon was 25 species for plants, 21 species for wild birds, 5 for mammalia, 10 for amphibians and reptiles and 16 species for insects. A site investigation for the second candidates was conducted to select the final candidate species. For the target areas, the habitats of the second candidates given by the literature and 46 potential areas in Seoul, which show good natural resistance, were finally selected. From the result of the site investigation, it was found that 14 species of plants, 19 species of wild birds, 2 species of mammalia, 4 species of amphibians and reptiles, and 16 species of insects were actually inhabiting Seoul; thus, the final 55 protected species were selected.