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Characterization of a Psychrophilic Metagenome Esterase EM2L8 and Production of a Chiral Intermediate for Hyperlipemia Drug (메타게놈유래의 저온성 에스터라제 EM2L8의 효소적 특성과 이를 활용한 고지혈증 치료제 키랄소재의 생산)

  • Jung, Ji-Hye;Choi, Yun-Hee;Lee, Jung-Hyun;Kim, Hyung-Kwoun
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.118-124
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    • 2009
  • Esterase EM2L8 gene isolated from deep sea sediment was expressed in Escherichia coli BL21 (DE3) and the esterase activity of the cell-free extract was assayed using p-nitrophenyl butyrate-spectrophotometric method. Its optimum temperature was $40-45^{\circ}C$ and 45% activity of the maximum activity was retained at $15^{\circ}C$. The activation energy at $15-45^{\circ}C$ was calculated to be 4.9 kcal/mol showing that esterase EM2L8 was a typical cold-adapted enzyme. Enzyme activity was maintained for 6 h and 4 weeks at $30^{\circ}C$ and $4^{\circ}C$, respectively. When each ethanol, methanol, and acetone was added to the reaction mixture to 15% concentration, enzyme activity was maintained. In the case of DMSO, enzyme activity was kept up to 40% concentration. (S)-4-Chloro-3-hydroxy butyric acid is a chiral intermediate for the synthesis of Atorvastatin, a hyperlipemia drug. When esterase EM2L8 (40 U) was added to buffer solution (1.2 mL, pH 9.0) containing ethyl-(R,S)-4-chloro-3-hydroxybutyrate (38 mM), it was hydrolyzed into 4-chloro-3-hydroxy butyric acid with a rate of $6.8\;{\mu}mole/h$. The enzyme hydrolyzed (S)-substrate more rapidly than (R)-substrate. When conversion yield was 80%, e.e.s value was 40%. When DMSO was added, hydrolysis rate increased to $10.4\;{\mu}mole/h$. The plots of conversion yield vs e.e.s in the presence or absence of DMSO were almost same, implying that the reaction enantioselectivity was not changed by the addition of DMSO. Taken together, esterase EM2L8 had high activity and stability at low temperatures as well as in various organic solvents/aqueous solutions. These properties suggested that it could be used as a biocatalyst in the synthesis of useful pharmaceuticals.

Monte-Carlo Simulations of Non-ergodic Solute Transport from Line Sources in Isotropic Mildly Heterogeneous Aquifers (불균질 등방 대수층 내 선형오염원으로부터 기원된 비에르고딕 용질 이동에 관한 몬테카를로 시뮬레이션)

  • Seo Byong-min
    • Journal of Soil and Groundwater Environment
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    • v.10 no.6
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    • pp.20-31
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    • 2005
  • Three dimensional Monte-Carlo simulations of non-ergodic transport of a lion-reactive solute plume by steady-state groundwater flow under a uniform mean velocity in isotropic heterogeneous aquifers were conducted. The log-normally distributed hydraulic conductivity, K(x), is modeled as a random field. Significant efforts are made to reduce tile simulation uncertainties. Ensemble averages of the second spatial moments of the plume and plume centroid variances were simulated with 1600 Monte Carlo runs for three variances of log K, ${\sigma}_Y^2=0.09,\;0.23$, and 0.46, and three dimensionless lengths of line plume sources normal to the mean velocity. The simulated second spatial moment and the plume centroid variance in longitudinal direction fit well to the first order theoretical results while the simulated transverse moments are generally larger than the first order results. The first order theoretical results significantly underestimated the simulated dimensionless transverse moments for the aquifers of large ${\sigma}_Y^2$ and large dimensionless time. The ergodic condition for the second spatial moments is far from reaching in all cases simulated, and transport In transverse directions may reach ergodic condition much slower than that in longitudinal direction. The evolution of the contaminant transported in a heterogeneous aquifer is not affected by the shape of the initial plume but affected mainly by the degree of the heterogeneity and the size of the initial plume.

Studies on the Changes of Nitrate, Nitrite Contents and the Viable Bacterial Population during Milk Storage (우유(牛乳) 저장중(貯藏中) 질산염(窒酸鹽) 및 아질산염(亞窒酸鹽)의 함량(含量)과 생균수(生菌數)의 변화(變化))

  • Choi, Jae-Chun;Kwon, Joong-Ho;Shin, Dae-Hyu;Yoon, Hyung-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.11 no.3
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    • pp.15-19
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    • 1982
  • The Changes of nitrate and nitrite contents at different storage temperatures (low tempenature : $4{\sim}7^{\circ}C$, high temperature: $28{\sim}30^{\circ}C$) were studied in relation to the viable bacterial population. The contents of nitrate and nitrite in milk were 1. 10 ppm and 0.03 ppm, respectively. When milk was storaged for 7 days at $28{\sim}30^{\circ}C$, nitrate concentration decreased to 0.3 to 0.6 ppm, whereas nitrite increased up to 0.13 to 0.08 ppm. However, during the storage at $4{\sim}7^{\circ}C$, their contents were little changed. The number of viable bacterial population in raw milk appeared $9{\times}10^6/ml$, and rapidly increased throughout 3 days storage at high temperature, but afterward it tended to decrease. This seemd to indicate that nitrate was reduced to nitrite by the nitrate reducing bacteia in milk, and the changes in the viable bacterial population were also related to the changes of pH and titratable acidity.

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Voluntary Motor Control Change after Gait Training in Patients with Spinal Cord Injury (척수신경손상 환자의 보행훈련 전.후의 능동적 근육제어의 변화)

  • 임현균;이동철;이영신;셔우드아더
    • Journal of Biomedical Engineering Research
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    • v.24 no.2
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    • pp.133-140
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    • 2003
  • In this study, muscle activity was measured using surface EMG (sEMG) during a voluntary maneuver (ankle dorsiflexion) in the supine position was compared pre and post gait training. Nine patients with incomplete spinal cord injury participated in a supported treadmill ambulation training (STAT), twenty minutes a day, five days a week for three months. Two tests, a gait speed test and a voluntary maneuver test, were made the same day, or at least the same week, pre and post gait training. Ten healthy subjects' data recorded using the same voluntary maneuvers were used for the reference. sEMG measured from ten lower limb muscles was used to observe the two features of amplitude and motor control distribution pattern, named response vector. The result showed that the average gait speed of patients increased significantly (p〈0.1) from 0.47$\pm$0.35 m/s to 0.68$\pm$0.52 m/s. In sEMG analysis, six out of nine patients showed a tendency to increase the right tibialis anterior activity during right ankle dorsiflexion from 109.7$\pm$148.5 $mutextrm{V}$ to 145.9$\pm$180.7 $mutextrm{V}$ but it was not significant (p〈0.055). In addition, only two patients showed increase of correlation coefficient and total muscle activity in the left fide during left dorsiflexion. Patients' muscle activity changes after gait training varied individually and generally depended on their muscle control abilities of the pre-STAT status. Response vector being introduced for quantitative analysis showed good Possibility to anticipate. evaluate, and/or guide patients with SCI, before and after gait training.

Life Cycle of Index Derivatives and Trading Behavior by Investor Types (주가지수 파생상품 Life Cycle과 투자자 유형별 거래행태)

  • Oh, Seung-Hyun;Hahn, Sang-Buhm
    • The Korean Journal of Financial Management
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    • v.25 no.2
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    • pp.165-190
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    • 2008
  • The degree of informational asymmetry relating to the expiration of index derivatives is usually increased as an expiration day of index derivatives approaches. The increase in the degree of informational asymmetry may have some effects on trading behavior of investors. To examine what the effects look like, 'life cycle of index derivatives' in this study is defined as three adjacent periods around expiration day: pre-expiration period(a week before the expiration day), post-expiration period(a week after the expiration day), and remaining period. It is inspected whether stock investor's trading behavior is changed according to the life cycle of KOSPI200 derivatives and what the reason of the changing behavior is. We have four results. First, trading behavior of each investor group is categorized into three patterns: ㄱ-pattern, L-pattern and U-pattern. The level of trading activity is low for pre-expiration period and normal for other periods in the ㄱ-pattern. L-pattern means that the level of trading activity is high for post-expiration period and normal for other periods. In the U-pattern, the trading activity is reduced for remaining period compared to other periods. Second, individual investors have ㄱ-pattern of trading large stocks according to the life cycle of KOSPI200 index futures while they show U-pattern according to the life cycle of KOSPI200 index options. Their trading behavior is consistent with the prediction of Foster and Viswanathan(1990)'s model for strategic liquidity investors. Third, trading pattern of foreign investors in relation to life cycle of index derivatives is partially explained by the model, but trading pattern of institutional investors has nothing to do with the predictions of the model.

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Comparison of Nutritional Composition in Korean Rices (쌀 품종별 백미와 현미의 영양성분 조성 비교)

  • 최정숙;안훈희;남희정
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.885-892
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    • 2002
  • The purpose of this research was to prove the excellency of unpolished (brown) rice with respect to nutritional composition and to compare chemical composition among six varieties of rice. Some nutritional composition in the Polished (well -milled) rice and unpolished (brown) rice of the six typical Korean varieties (Chucheong, Hwasung, Odae, Ilmi, Dongjin, Ilpum) were determined. Most of nutrients in the brown rice were much greater than those in the milled rice and there was significant difference among rice varieties in some chemical compositions. The major minerals of milled rices were Ca, 6~15 mg%;P, 91~125mg%;Fe, 0.3~1.2mg%; Na, 10~14mg%;K, 106~205 mg%; Zn, 1.0~1.8mg%; and Mg, 32~58 mg%. The mineral contents of brown rices wre Ca, 4~11mg%;P, 92~286mg%;Fe, 1.3~1.9mg%;Na, 12~15mg%;K, 243~320mg%;Zn, 1.5~2.3mg%;Mg, 112~140mg%. Major fatty acids in six rice varieties were linoleic acid, oleic acid and palmitic acid, which comprised of about 93% of total fatty acid. Amino acid analysis showed that aspartic acid, glutamic acid, arginine and leucine were major amino acids, whereas histidine, threonine and tryptophan were minor ones of rice. Most ammo acid contents was higher in Ilmi than the other varieties. The contents of vitamin in brown rices were considerably higher than those in well-milled rice. There was not significant difference in total dietary fiber among rice varieties : 1.1~1.2% for milled rice,3.2~3.5% for brown rice.

Determination of Tumor Boundaries on CT Images Using Unsupervised Clustering Algorithm (비교사적 군집화 알고리즘을 이용한 전산화 단층영상의 병소부위 결정에 관한 연구)

  • Lee, Kyung-Hoo;Ji, Young-Hoon;Lee, Dong-Han;Yoo, Seoung-Yul;Cho, Chul-Koo;Kim, Mi-Sook;Yoo, Hyung-Jun;Kwon, Soo-Il;Chun, Jun-Chul
    • Journal of Radiation Protection and Research
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    • v.26 no.2
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    • pp.59-66
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    • 2001
  • It is a hot issue to determine the spatial location and shape of tumor boundary in fractionated stereotactic radiotherapy (FSRT). We could get consecutive transaxial plane images from the phantom (paraffin) and 4 patients with brain tumor using helical computed tomography(HCT). K-means classification algorithm was adjusted to change raw data pixel value in CT images into classified average pixel value. The classified images consists of 5 regions that ate tumor region (TR), normal region (NR), combination region (CR), uncommitted region (UR) and artifact region (AR). The major concern was how to separate the normal region from tumor region in the combination area. Relative average deviation analysis was adjusted to alter average pixel values of 5 regions into 2 regions of normal and tumor region to define maximum point among average deviation pixel values. And then we drawn gross tumor volume (GTV) boundary by connecting maximum points in images using semi-automatic contour method by IDL(Interactive Data Language) program. The error limit of the ROI boundary in homogeneous phantom is estimated within ${\pm}1%$. In case of 4 patients, we could confirm that the tumor lesions described by physician and the lesions described automatically by the K-mean classification algorithm and relative average deviation analyses were similar. These methods can make uncertain boundary between normal and tumor region into clear boundary. Therefore it will be useful in the CT images-based treatment planning especially to use above procedure apply prescribed method when CT images intermittently fail to visualize tumor volume comparing to MRI images.

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Effect of Radiation Dose for Radiotherapy on Ovarian Follicle Atresia in Rat (치료 방사선량이 쥐의 난포 퇴축에 미치는 영향)

  • Lee, Won-Jeong;Seon, Jong-Ryul;Yoo, Se-Jong;Ahn, Bong-Seon
    • Journal of Radiation Protection and Research
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    • v.37 no.4
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    • pp.208-212
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    • 2012
  • In previous studies, ovarian follicle in rat has been used a higher radiation dose than that for cancer radiotherapy in clinical practice. The aim of this study was to evaluate the effect of radiation dose used for cancer radiotherapy on ovarian follicle atresia in rat. Mice of 4-week-old female were whole body irradiated with 2 cGy or 2 Gy (Mevatron 67, Siemens, Germany) and sacrificed by cervical dislocation. Ovaries were collected at 24 hours after irradiation to observe the degree of follicular atresia. Ovaries were fixed in neutral formaldehyde solution for 24 hours and embedded with paraffin. Cutted in $5{\mu}m$ thickness with microtome and stained with hematoxylin and eosin (H&E) and TUNEL immunohistochemical stain, and examined histologically under a light microscope. All data were presented as mean ${\pm}SD$, calculating the ratio of normal or atretic follicles to total ovarian follicles. Statistical analysis was performed by the Mann Whitney test using the SPSS ver 19.0. Ratio of atretic to total follicles of 2 Gy group was significantly higher than control or 2 cGy groups (p<0.05). Ratio of normal to total follicles of 2 Gy group was significantly lower than control group in preantral follicle (64.0 vs. 87.7, p=0.027). Ratio of normal to total follicles of 2 cGy group was significantly increased more than control or 2 Gy groups in antral follicle, and there were no significant difference between control and 2 Gy groups (p=0.522). Radiation dose of 2 Gy for cancer radiotherapy have a significant effect on ovarian follicle atresia in rat.

Performance Test of Portable Hand-Held HPGe Detector Prototype for Safeguard Inspection (안전조치 사찰을 위한 휴대형 HPGe 검출기 시제품 성능평가 실험)

  • Kwak, Sung-Woo;Ahn, Gil Hoon;Park, Iljin;Ham, Young Soo;Dreyer, Jonathan
    • Journal of Radiation Protection and Research
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    • v.39 no.1
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    • pp.54-60
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    • 2014
  • IAEA has employed various types of radiation detectors - HPGe, NaI, CZT - for accountancy of nuclear material. Among them, HPGe has been mainly used in verification activities required for high accuracy. Due to its essential cooling component(a liquid-nitrogen cooling or a mechanical cooling system), it is large and heavy and needs long cooling time before use. New hand-held portable HPGe has been developed to address such problems. This paper deals with results of performance evaluation test of the new hand-held portable HPGe prototype which was used during IAEA's inspection activities. Radioactive spectra obtained with the new portable HPGe showed different characteristics depending on types and enrichments of nuclear materials inspected. Also, Gamma-rays from daughter radioisotopes in the decay series of $^{235}U$ and $^{238}U$ and characteristic x-rays from uranium were able to be remarkably separated from other peaks in the spectra. A relative error of enrichment measured by the new portable HPGe was in the range of 9 to 27%. The enrichment measurement results didn't meet partially requirement of IAEA because of a small size of a radiation sensing material. This problem might be solved through a further study. This paper discusses how to determine enrichment of nuclear material as well as how to apply the new hand-held portable HPGe to safeguard inspection. There have been few papers to deal with IAEA inspection activity in Korea to verify accountancy of nuclear material in national nuclear facilities. This paper would contribute to analyzing results of safeguards inspection. Also, it is expected that things discussed about further improvement of a radiation detector would make contribution to development of a radiation detector in the related field.

Quality Characteristics of Fresh Noodles With Perilla Leaves (들깻잎을 첨가한 생면의 품질특성)

  • Kim, Chang-Yeol;Choi, Sang-Ho;Kim, Jung-Soo
    • Culinary science and hospitality research
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    • v.18 no.2
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    • pp.182-196
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    • 2012
  • This study investigates the optimal application of perilla leaves to fresh noodles in terms of a variety of beneficial effects and functions such as the anti-microbial function of the leaves. First, we measured the water contents of the fresh noodles and found that the ones added with 7% lyophilized perilla leaf extract showed the maximum water contents whereas the control noodles without perilla leaf extract the minimum. Increasing amount of raw perilla leaf extract or lysophilzed perilla leaves in the fresh noodles elevated the L value and significantly reduced the A value. The mass and volume of the fresh noodles were gradually decreased with raw perilla leaves added. Addition of raw perilla leaf extract and lyophilized perilla leaves decreased the turbidity of the fresh noodles. Hardness of the fresh noodles was increased by the addition of lyophilized Perilla leaves. Next, we tested the effect of the addition of sesame leaves on microbial growth. No microbes were found in the fresh noodles in the absence or presence of sesame leaves at day 0. When the noodles were stored for 3 days, the greatest number of bacteria was detected in the noodles without perilla leaves while addition of perilla leaves lowered the amount of bacteria in the noodles. We then performed the sensory test. For the raw perilla leaf extract addition, the noodles with 9% of extract exhibited the highest in appearance, flavor, color (6.47), texture (6.60), and overall acceptability (7.67). Texture was the highest in the ones with 3% (6.87) and 5% (6.20) of extracts added. Overall acceptability (7.07) was the best when 3% perilla leaves were added. Overall, 9% addition of raw perilla leaf extract or 3% addition of lysophilized perilla leaves showed optimal tastes.

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