• Title/Summary/Keyword: Kinetic study

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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.

Metabolic Adjustments of Lactate Dehydrogenase Isozymes to the Environmental Temperature in Bluegill (Lepomis macrochirus) (환경온도에 대한 파랑볼우럭(Lepomis macrochirus) 젖산탈수소효소 동위효소들의 대사조절)

  • Ku, Bora;Cho, Sung Kyu;Yum, Jung Joo
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1105-1112
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    • 2016
  • The aim of this study was to examine the metabolic adjustment of lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes to the environmental temperature in bluegill (Lepomis macrochirus). This study included three groups of bluegill collected in April (group Ⅰ), May (group Ⅱ), and September (group Ⅲ). The LDH activities of skeletal muscle, heart, and brain tissues were higher in group Ⅲ than in groups Ⅰ and Ⅱ. The citrate synthase (EC 4.1.3.7, CS) activity was higher in skeletal muscle but lower in heart and brain tissues of group Ⅱ as compared to group Ⅰ. In contrast, the CS activity was lower in skeletal muscle and higher in heart and brain tissues in group Ⅲ than in group Ⅱ. Furthermore, the LDH/CS activity ratio was higher in the skeletal muscle and brain in group Ⅲ than in groups Ⅰ and Ⅱ. Accordingly, anaerobic metabolism was increased in group Ⅲ. LDH A4, A2B2, and B4 isozymes were expressed in skeletal muscle, heart, liver, and brain tissues. The LDH C hybrid was detected in brain tissue. The LDH A4 isozyme was successfully purified by affinity chromatography. The molecular weight of the purified LDH A4 isozyme was 136 kDa and its optimal pH for enzymatic activity was 8.0. The KmPYR values of LDH in skeletal muscle were 0.161-0.227 mM using pyruvate as a substrate. These kinetic properties of LDH in skeletal muscle are consistent with the fact that bluegill is a cold-adapted species. These results may be useful for predicting the habitat use of this fish.

Removal Characteristics of Sulfonamide Antibiotic Compounds in Biological Activated Carbon Process (생물활성탄 공정에서의 Sulfonamide계 항생물질 제거특성)

  • Son, Hee-Jong;Jung, Jong-Moon;Roh, Jae-Soon;Yu, Pyung-Jong
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.2
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    • pp.96-101
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    • 2009
  • In this study, the effects of three different biological activated carbon (BAC) materials (each coal, coconut and wood based activated carbons), empty bed contact time (EBCT) and water temperature on the removal of sulfonamide 5 species in BAC filters were investigated. Experiments were conducted at three water temperatures (5, 15 and $25^{\circ}C$) and four EBCTs (5, 10, 15 and 20 min). The results indicated that coal based BAC retained more attached bacterial biomass on the surface of the activated carbon than the other BACs, increasing EBCT or increasing water temperature increased the sulfonamide 5 species removal in BAC columns. In the coal-based BAC columns, sulfachloropyridazine (SCP), sulfamethazine (SMT) and sulfathiazole (STZ) removal efficiencies were 30~80% and sulfadimethoxine (SDM), sulfamethoxazole (SMX) removal efficiencies were 18~70% for 5~20 min EBCT at $25^{\circ}C$. The kinetic analysis suggested a first-order reaction model for sulfonamide 5 species removal at various water temperatures (5~$25^{\circ}C$). The pseudo-first-order reaction rate constants and half-lives were also calculated for sulfonamide 5 species removal at 5~$25^{\circ}C$. The reaction rate and half-lives of sulfonamide 5 species ranging from 0.0094~0.0718 $min^{-1}$ and 9.7 to 73.7 min various water temperaturs and EBCTs in this study could be used to assist water utilities in designing and operating BAC filters for sulfonamide antibiotic compounds removal.

Correlation Analysis of Diffusion Metrics (FA and ADC) Values Derived from Diffusion Tensor Magnetic Resonance Imaging in Breast Cancer (유방암의 확산텐서 자기공명 영상에서 유도된 확산 지표(FA, ADC) 값의 연관성 분석)

  • Lee, Jae-Heun;Lee, Hyo-Yeong
    • Journal of the Korean Society of Radiology
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    • v.12 no.6
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    • pp.755-762
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    • 2018
  • The purpose of this study was to compare the FA(faractional anisotropy) and ADC(apparent diffusion coefficient) values, which were derived from diffusion tensor imaging in breast cancer patients. The diffusion gradient used in this study was derived from quantitative diffusion indices using 20 directions(b-value, 0 and $1,000s/mm^2$). Quantitative analysis was analyzed using Pearson's correction and qualitative analysis using for correction coefficients. As a result, $FA_{min}$, $FA_{mean}$ and $FA_{max}$ were $0.098{\pm}0.065$, $0.302{\pm}0.142$ and $0.634{\pm}0.236$, respectively(p > 0.05). The $ADC_{min}$, $ADC_{mean}$ and $ADC_{max}$ were $0.741{\pm}0.403$, $1.095{\pm}0.394$ and $1.530{\pm}0.447$, respectively(p > 0.05). The $FA_{min}$, $FA_{mean}$, and $FA_{max}$ mean values were $0.132{\pm}0.050$, $0.418{\pm}0.094$, and $0.770{\pm}0.164$ for Category 6 and Kinetic Curve Pattern III, respectively. $ADC_{min}$, $ADC_{mean}$, and $ADC_{max}$ were $0.753{\pm}0.189$, $1.120{\pm}0.236$, and $1.615{\pm}0.372$, respectively. Quantitative analysis showed negative correlation between $ADC_{mean}-FA_{mean}$ and $ADC_{max}-FA_{max}$(p = 0.001, 0.003). The qalitative analysis showed ADC 0.628(p = 0.001), FA 0.620(p = 0.001) in the internal evaluations, ADC 0.677(p = 0.001), FA 0.695(p = 0.001) in external evaluations. In conclusion, based on the morphological examination, time to signal intensity graph is the form of wash-out(pattern III) in the dynamic contrast enhance examination, As a result, the $ADC_{mean}$ $1.120{\pm}0.236$ and $FA_{mean}$ values were $0.032{\pm}0.142$ with a negative correlation (Y=1.44-1.12X). Therefore, If we understand the shape of time to signal intensity graph and the relationship between ADC and FA, It will be a criterion for distinguishing malignant diseases in breast cancer.

Water Digital Twin for High-tech Electronics Industrial Wastewater Treatment System (II): e-ASM Calibration, Effluent Prediction, Process selection, and Design (첨단 전자산업 폐수처리시설의 Water Digital Twin(II): e-ASM 모델 보정, 수질 예측, 공정 선택과 설계)

  • Heo, SungKu;Jeong, Chanhyeok;Lee, Nahui;Shim, Yerim;Woo, TaeYong;Kim, JeongIn;Yoo, ChangKyoo
    • Clean Technology
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    • v.28 no.1
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    • pp.79-93
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    • 2022
  • In this study, an electronics industrial wastewater activated sludge model (e-ASM) to be used as a Water Digital Twin was calibrated based on real high-tech electronics industrial wastewater treatment measurements from lab-scale and pilot-scale reactors, and examined for its treatment performance, effluent quality prediction, and optimal process selection. For specialized modeling of a high-tech electronics industrial wastewater treatment system, the kinetic parameters of the e-ASM were identified by a sensitivity analysis and calibrated by the multiple response surface method (MRS). The calibrated e-ASM showed a high compatibility of more than 90% with the experimental data from the lab-scale and pilot-scale processes. Four electronics industrial wastewater treatment processes-MLE, A2/O, 4-stage MLE-MBR, and Bardenpo-MBR-were implemented with the proposed Water Digital Twin to compare their removal efficiencies according to various electronics industrial wastewater characteristics. Bardenpo-MBR stably removed more than 90% of the chemical oxygen demand (COD) and showed the highest nitrogen removal efficiency. Furthermore, a high concentration of 1,800 mg L-1 T MAH influent could be 98% removed when the HRT of the Bardenpho-MBR process was more than 3 days. Hence, it is expected that the e-ASM in this study can be used as a Water Digital Twin platform with high compatibility in a variety of situations, including plant optimization, Water AI, and the selection of best available technology (BAT) for a sustainable high-tech electronics industry.

Lower Limbs Muscle Comparative Research for Verification Effect of Rehabilitation Training Program of Total Hip Arthroplasty (재활운동 프로그램에 참가한 엉덩인공관절 수술자의 하지근력 변화에 대한 비교연구)

  • Jin, Young-Wan
    • Journal of Life Science
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    • v.20 no.4
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    • pp.543-548
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    • 2010
  • The purpose of this study was to examine the differences in kinetics between 6 months of rehabilitation training and 12 months of rehabilitation training after total hip arthroplasty. 10 unilateral THA participants performed kinetic tests. Three dimensional kinematics and hip flexors and abductors electromyography (EMG) were collected during each trial. T-test was used for statistical analysis (p<0.05). There was no significant difference in EMG data between the two groups, but the mean comparison EMG data was higher in the 12 months rehabilitation training group than the 6 months rehabilitation training group. The moment value was found with motion-dependent interaction analyzing method which was used by Feltner and Dapena. There was no significant difference between moment values of the two groups. There was no significant difference between ground reaction forces of the two groups; however, there were some differences shown in Fz (vertical reaction force) between the two groups ($892{\pm}104\;N$, $820{\pm}87\;N$). The first peak impact force was about 9% lower in the 12 months group compared to the 6 months group. The second peak active force was nearly equal between the two groups. More research is necessary to determine exactly what constitutes optimal rehabilitation training biomechanics for patients with total hip arthroplasty.

Kinetic and Statistical Analysis of Adsorption and Photocatalysis on Sulfamethoxazole Degradation by UV/$TiO_2$/HAP System (UV/$TiO_2$/HAP 시스템에서 Sulfamethoxazole의 흡착과 광촉매반응에 대한 동역학적 및 통계적 해석)

  • Chun, Suk-Young;Chang, Soon-Woong
    • Journal of the Korean GEO-environmental Society
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    • v.13 no.5
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    • pp.5-12
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    • 2012
  • Antibiotics have been considered emerging compounds due to their continuous input and persistence in environment. Due to the limited biodegradability and widespread use of these antibiotics, an incomplete removal is attained in conventional wastewater treatment plants and relative large quantities are released into the environment. In this study, it was determined the adsorption and photocatalysis kinetics of antibiotics (Sulfamethoxazole, SMX) with various catalyst (Titanium dioxide; $TiO_2$, Hydroxyapatite; HAP) conditions under UV/$TiO_2$/HAP system. In addition, the statistical analysis of response surface methods (RSM) was used to determine the effects of operating parameters on UV/$TiO_2$/HAP system. $TiO_2$/HAP adsorbent were found to follow the pseudo second order reaction in the adsorption. In the result of applied intrapaticle diffusion model, the constants of reaction rate were $TiO_2$=$0.064min^{-1}$, HAP=$0.2866min^{-1}$ and $TiO_2$/HAP=$0.3708min^{-1}$, respectively.The result of RSM, term of regression analysis in analysis of variance (ANOVA) showed significantly p-value (p<0.05) and high coefficients for determination values($R^2$=96.2%, $R^2_{Adj}$=89.3%) that allowed satisfactory prediction of second order regression model. And the estimated optimal conditions for Y(Sulfamethoxazole removal efficiency, %) were $x_1$(initial concentration of Sulfamethoxazole)=-0.7828, $x_2$(amount of catalyst)=0.9974 and $x_3$(reation time)=0.5738 by coded parameters, respectively. According to the result of intraparticle diffusion model and photocatalysis experiments, it was shown that the $TiO_2$/HAP was more effective system than conventional AOPs(advanced oxidation processes, UV/$TiO_2$ system).

Study on Pretreatment Methods to Prevent Tissue Softening of Heated Onion (가열 양파의 조직 연화 방지를 위한 전처리 방법에 관한 연구)

  • Choi, Jun-Bong;Cho, Won-Il
    • Korean Journal of Food Science and Technology
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    • v.47 no.1
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    • pp.56-62
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    • 2015
  • Various pretreatment methods were evaluated to prevent tissue softening of heated onion. Changes in onion tissue firmness during heating were explained by 3-mechanism model consisting of texture hardening at low temperature ($60-80^{\circ}C$) and substrate softening at high temperature. Preheating of onion in a $Ca^{2+}$-containing solution significantly improved its texture after high-temperature heating. The improvement of firmness by preheating at low temperature was related to the formation of strong cross-linking between carboxyl groups and $Ca^{2+}$ by the action of pectin methylesterase in onion. The highest firmness was obtained by pre-heating at $70^{\circ}C$ for 120 min in 0.5% calcium solution. This result was supported by chemical analysis showing that the amount of bound calcium was the highest at $70^{\circ}C$. Further investigation should be carried out to establish the optimal conditions to prevent the softening of various vegetables.

A Biomechanical Analysis of Judo's Kuzushi(balance-breaking) Motion (유도 팔방기울이기 동작의 생체역학적 특성 분석)

  • Kim, Sung-Sup;Kim, Eui-Hwan;Kim, Tae-Whan
    • Korean Journal of Applied Biomechanics
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    • v.17 no.2
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    • pp.207-216
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    • 2007
  • The purpose of this study was to biomechanical analysis Judo's Kuzushi throwing motion in order to increase the effectiveness of Nage-waja(throwing technique). The Tori was a Judo player with 18 years experience(4th degree) while the Uke was a player with 2 years experience(1st degree). The kinematic data was captured using the Vicon motion system (7 cameras) and the kinetics were recorded by force plates(2 AMTI). The following were the results; While leaning to the front the subject's trunk's angle was $14.5^{\circ}$, the lower limbs angle was $23.8^{\circ}$, knee angle was $179.6^{\circ}$ and the vertical reaction of the left leg was 325.42N(BW 0.34) and the right leg was 233.7N(BW 0.47). While leaning back the subject's trunk's angle was $11.3^{\circ}$, the lower limbs angle was $4.1^{\circ}$, knee angle was $1761^{\circ}$ and the vertical reaction of the left leg was 299.53N(BW 0.43) and the right leg was 441.7N(BW 0.64). While leaning to the left the subject's trunk's angle was $30.8^{\circ}$, the lower limbs angle was $2.7^{\circ}$, knee angle was $175.2^{\circ}$ and the vertical reaction of the left leg was 711N(BW 1.03) and the right leg was 9.2N(BW 0.01). While leaning to the right the subject's trunk's angle was $36.5^{\circ}$, the lower limbs angle was $10.4^{\circ}$, knee angle was $175.2^{\circ}$ and the vertical reaction of the left leg was 13.2N(BW 0.02) and the right leg was 694.7N(BW 1.01). While leaning to the left front corner the subject's trunk's angle was $19.8^{\circ}$ (front) and $15.1^{\circ}$ (left), the lower limbs angle was $17.8^{\circ}$ (front) and $2.4^{\circ}$ (left), knee angle was $177.8^{\circ}$ (front) and $173.9^{\circ}$(left), and the vertical reaction of the left leg was 547.4N(BW 0.8) and the right leg was 117.8N(BW 0.17). While leaning to the right front corner the subject's trunk's angle was $15.4^{\circ}$ (front) and $17.7^{\circ}$ (right), the lower limbs angle was $21.1^{\circ}$, (front) and $5.7^{\circ}$ (right), knee angle was $175.5^{\circ}$ (front) and $178.9^{\circ}$(right), and the vertical reaction of the left leg was 53N(BW 0.08) and the right leg was 622.4N(BW 09). While leaning to the left rear corner the subject's trunk's angle was $9.2^{\circ}$ (back) and $13.8^{\circ}$ (left), the lower limbs angle was $2^{\circ}$, (back) and $5.7^{\circ}$ (left), knee angle was $175.5^{\circ}$ (back) and $172.8^{\circ}$(left), and the vertical reaction of the left leg was 698.2N(BW 1.02) and the right leg was 49.6N(BW 0.07). While leaning to the right rear corner the subject's trunk's angle was $8.9^{\circ}$ (back) and $19.6^{\circ}$ (right), the lower limbs angle was ${0.6^{\circ}}_"$ (back) and $3.1^{\circ}$ (right), knee angle was $174.6^{\circ}$ (back) and $175.6^{\circ}$(right), and the vertical reaction of the left leg was 7.2N(BW 0.01) and the right leg was 749.4N(BW 1.09). It was observed that during the Judo motion Kuzushii the range of the COM varied from $26.5{\sim}39.9cm$. It was concluded that the upper body leaned further than the lower body as there was knee extension. There was high left leg reaction forces while leaning to the left and likewise for the right side. It was therefore deduced that the Kuzushi was a more effective throwing technique for the left side.

Prediction of Growth of Escherichia coli O157 : H7 in Lettuce Treated with Alkaline Electrolyzed Water at Different Temperatures

  • Ding, Tian;Jin, Yong-Guo;Rahman, S.M.E.;Kim, Jai-Moung;Choi, Kang-Hyun;Choi, Gye-Sun;Oh, Deog-Hwan
    • Journal of Food Hygiene and Safety
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    • v.24 no.3
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    • pp.232-237
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    • 2009
  • This study was conducted to develop a model for describing the effect of storage temperature (4, 10, 15, 20, 25, 30 and $35^{\circ}C$) on the growth of Escherichia coli O157 : H7 in ready-to-eat (RTE) lettuce treated with or without (control) alkaline electrolyzed water (AIEW). The growth curves were well fitted with the Gompertz equation, which was used to determine the specific growth rate (SGR) and lag time (LT) of E. coli O157 : H7 ($R^2$ = 0.994). Results showed that the obtained SGR and LT were dependent on the storage temperature. The growth rate increased with increasing temperature from 4 to $35^{\circ}C$. The square root models were used to evaluate the effect of storage temperature on the growth of E. coli O157 : H7 in lettuce samples treated without or with AIEW. The coefficient of determination ($R^2$), adjusted determination coefficient ($R^2_{Adj}$), and mean square error (MSE) were employed to validate the established models. It showed that $R^2$ and $R^_{Adj}$ were close to 1 (> 0.93), and MSE calculated from models of untreated and treated lettuce were 0.031 and 0.025, respectively. The results demonstrated that the overall predictions of the growth of E. coli O157: H7 agreed with the observed data.