• Journal of Microbiology and Biotechnology
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• 제24권7호
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• pp.925-935
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• 2014

A cold-adapted carbohydrate esterase, CEST, belonging to the carbohydrate esterase family 6, was cloned from Microbulbifer thermotolerans DAU221. CEST was composed of 307 amino acids with the first 22 serving as a secretion signal peptide. The calculated molecular mass and isoelectric point of the mature enzyme were 31,244 Da and pH 5.89, respectively. The catalytic triad consisted of residues Ser37, Glu192, and His281 in the conserved regions: GQSNMXG, QGEX(D/N), and DXXH. The three-dimensional structure of CEST revealed that CEST belongs to the ${\alpha}/{\beta}$-class of protein consisted of a central six-stranded ${\beta}$-sheet flanked by eight ${\alpha}$-helices. The recombinant CEST was purified by His-tag affinity chromatography and the characterization showed its optimal temperature and pH were $15^{\circ}C$ and 8.0, respectively. Specifically, CEST maintained up to 70% of its enzyme activity when preincubated at $50^{\circ}C$ or $60^{\circ}C$ for 6 h, and 89% of its enzyme activity when preincubated at $70^{\circ}C$ for 1 h. The results suggest CEST belongs to group 3 of the cold-adapted enzymes. The enzyme activity was increased by $Na^+$ and $Mg^{2+}$ ions but was strongly inhibited by $Cu^+$ and $Hg^{2+}$ ions, at all ion concentrations. Using p-nitrophenyl acetate as a substrate, the enzyme had a $K_m$ of 0.278 mM and a $k_{cat}$ of $1.9s^{-1}$. Site-directed mutagenesis indicated that the catalytic triad (Ser37, Glu192, and His281) and Asp278 were essential for the enzyme activity.

• 한국의학물리학회지:의학물리
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• 제29권1호
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• pp.29-41
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• 2018

The objective of this study was to evaluate the chemical exchange saturation transfer (CEST) effect of amino acids and neurotransmitters, which exist in the human brain, depending on the concentration, pH, and amplitude of the saturation radiofrequency field. Phantoms were developed with asparagine (Asn), ${\gamma}-aminobutyric$ acid (GABA), glutamate (Glu), glycine (Gly), and myoinositol (MI). Each chemical had three different concentrations of 10, 30, and 50 mM and three different pH values of 5.6, 6.2, and 7.4. Full Z-spectrum CEST images for each phantom were acquired with a continuous-wave radiofrequency (RF) saturation pulse with two different $B_1$ amplitudes of $2{\mu}T$ and $4{\mu}T$ using an animal 9.4T MRI system. A voxel-based CEST asymmetry was mapped to evaluate exchangeable protons based on amide (-NH), amine ($-NH_2$), and hydroxyl (-OH) groups for the five target molecules. For all target molecules, the CEST effect was increased with increasing concentration and B1 amplitude; however, the CEST effect with varying pH displayed a different trend depending on the characteristics of the molecule. On CEST asymmetric maps, Glu and MI were well visualized around 3.0 and 0.9 ppm, respectively, and were well separated macroscopically at a pH of 7.4. The exchange rates of Asn, Glu, BABA, and Gly usually decreased with increasing pH. The CEST effect was dependent on the concentration, acidity of the target molecules, and B1 amplitude of the saturation RF pulse. The CEST effect for Asn can be observed in a 9.4T MRI system. The results of this study are based on applying the CEST technique in patients with neurodegenerative diseases when proteins in the brain are increased with disease progression.

• Korean Journal of Radiology
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• 제22권5호
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• pp.770-781
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• 2021

Objective: Chemical exchange-dependent saturation transfer (CEST) MRI is sensitive for detecting solid-like proteins and may detect changes in the levels of mobile proteins and peptides in tissues. The objective of this study was to evaluate the characteristics of chemical exchange proton pools using the CEST MRI technique in patients with dementia. Materials and Methods: Our institutional review board approved this cross-sectional prospective study and informed consent was obtained from all participants. This study included 41 subjects (19 with dementia and 22 without dementia). Complete CEST data of the brain were obtained using a three-dimensional gradient and spin-echo sequence to map CEST indices, such as amide, amine, hydroxyl, and magnetization transfer ratio asymmetry (MTR_asym) values, using six-pool Lorentzian fitting. Statistical analyses of CEST indices were performed to evaluate group comparisons, their correlations with gray matter volume (GMV) and Mini-Mental State Examination (MMSE) scores, and receiver operating characteristic (ROC) curves. Results: Amine signals (0.029 for non-dementia, 0.046 for dementia, p = 0.011 at hippocampus) and MTR_asym values at 3 ppm (0.748 for non-dementia, 1.138 for dementia, p = 0.022 at hippocampus), and 3.5 ppm (0.463 for non-dementia, 0.875 for dementia, p = 0.029 at hippocampus) were significantly higher in the dementia group than in the non-dementia group. Most CEST indices were not significantly correlated with GMV; however, except amide, most indices were significantly correlated with the MMSE scores. The classification power of most CEST indices was lower than that of GMV but adding one of the CEST indices in GMV improved the classification between the subject groups. The largest improvement was seen in the MTR_asym values at 2 ppm in the anterior cingulate (area under the ROC curve = 0.981), with a sensitivity of 100 and a specificity of 90.91. Conclusion: CEST MRI potentially allows noninvasive image alterations in the Alzheimer's disease brain without injecting isotopes for monitoring different disease states and may provide a new imaging biomarker in the future.

• 한국의학물리학회지:의학물리
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• 제28권4호
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• pp.135-143
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• 2017

Chemical Exchange Saturation Transfer (CEST) imaging is a method to detect solutes based on the chemical exchange of mobile protons with water. The solute protons exchange with three different patterns, which are fast, slow, and intermediate rates. The CEST contrast can be obtained from the exchangeable protons, which are hydroxyl protons, amine protons, and amide protons. The CEST MR imaging is useful to evaluate tumors, strokes, and other diseases. The purpose of this study is to review the mathematical model for CEST imaging and for measurement of the chemical exchange rate, and to measure the chemical exchange rate using a 3T MRI system on several amino acids. We reviewed the mathematical models for the proton exchange. Several physical models are proposed to demonstrate a two-pool, three-pool, and four-pool models. The CEST signals are also evaluated by taking account of the exchange rate, pH and the saturation efficiency. Although researchers have used most commonly in the calculation of CEST asymmetry, a quantitative analysis is also developed by using Lorentzian fitting. The chemical exchange rate was measured in the phantoms made of asparagine (Asn), glutamate (Glu), ${\gamma}-aminobutyric$ acid (GABA), glycine (Gly), and myoinositol (MI). The experiment was performed at a 3T human MRI system with three different acidity conditions (pH 5.6, 6.2, and 7.4) at a concentration of 50 mM. To identify the chemical exchange rate, the "lsqcurvefit" built-in function in MATLAB was used to fit the pseudo-first exchange rate model. The pseudo-first exchange rate of Asn and Gly was increased with decreasing acidity. In the case of GABA, the largest result was observed at pH 6.2. For Glu, the results at pH 5.6 and 6.2 did not show a significant difference, and the results at pH 7.4 were almost zero. For MI, there was no significant difference at pH 5.6 or 7.4, however, the results at pH 6.2 were smaller than at the other pH values. For the experiment at 3T, we were only able to apply 1 s as the maximum saturation duration due to the limitations of the MRI system. The measurement of the chemical exchange rate was limited in a clinical 3T MRI system because of a hardware limitation.

• 한국의학물리학회지:의학물리
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• 제30권4호
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• pp.139-149
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• 2019

Purpose: With neurodegeneration, the signal intensity of the cerebrospinal fluid (CSF) in the brain increases. The objective of this study was to evaluate chemical exchange saturation transfer (CEST) signals with and without the contribution of CSF signals in elderly human brains using two different 3T magnetic resonance imaging (MRI) sequences Methods: Full CEST signals were acquired in ten subjects (Group I) with a three-dimensional (3D)-segmented gradient-echo echo-planar imaging (EPI) sequence and in ten other subjects (Group II) with a 3D gradient and spin-echo (GRASE) sequence using two different 3T MRI systems. The segmented tissue compartments of gray and white matter were used to mask the CSF signals in the full CEST images. Two sets of magnetization transfer ratio asymmetry (MTR_asym) maps were obtained for each offset frequency in each subject with and without masking the CSF signals (masked and unmasked conditions, respectively) and later compared using paired t-tests. Results: The region-of-interest (ROI)-based analyses showed that the MTR_asym values for both the 3D-segmented gradient-echo EPI and 3D GRASE sequences were altered under the masked condition compared with the unmasked condition at several ROIs and offset frequencies. Conclusions: Depending on the imaging sequence, the MTR_asym values can be overestimated for some areas of the elderly human brain when CSF signals are unmasked. Therefore, it is necessary to develop a method to minimize this overestimation in the case of elderly patients.

• 한국자기학회:학술대회 개요집
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• 한국자기학회 2015년도 임시총회 및 하계학술연구발표회
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• pp.59-59
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• 2015

• Korean Journal of Radiology
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• 제21권5호
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• pp.588-597
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• 2020

Objective: To investigate the value of combined chemical exchange saturation transfer (CEST) and conventional magnetization transfer imaging (MT) in detecting metabolic and structural changes of renal fibrosis in rats with unilateral ureteral obstruction (UUO) at 3T MRI. Materials and Methods: Thirty-five Sprague-Dawley rats underwent UUO surgery (n = 25) or sham surgery (n = 10). The obstructed and contralateral kidneys were evaluated on days 1, 3, 5, and 7 after surgery. After CEST and MT examinations, ¹⁸F-labeled fluoro-2-deoxyglucose positron emission tomography was performed to quantify glucose metabolism. Fibrosis was measured by histology and western blots. Correlations were compared between asymmetrical magnetization transfer ratio at 1.2 ppm (MTR_asym(1.2ppm)) derived from CEST and maximum standard uptake value (SUV_max) and between magnetization transfer ratio (MTR) derived from MT and alpha-smooth muscle actin (α-SMA). Results: On days 3 and 7, MTR_asym(1.2ppm) and MTR of UUO renal cortex and medulla were significantly different from those of contralateral kidneys (p < 0.05). On day 7, MTR_asym(1.2ppm) and MTR of UUO renal cortex and medulla were significantly different from those of sham-operated kidneys (p < 0.05). The MTR_asym(1.2ppm) of UUO renal medulla was fairly negatively correlated with SUV_max (r = -0.350, p = 0.021), whereas MTR of UUO renal medulla was strongly negatively correlated with α-SMA (r = -0.744, p < 0.001). Conclusion: CEST and MT could provide metabolic and structural information for comprehensive assessment of renal fibrosis in UUO rats in 3T MRI and may aid in clinical monitoring of renal fibrosis in patients with chronic kidney disease.

• 한국통신학회논문지
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• 제42권5호
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• pp.1069-1076
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• 2017

C-ITS(Cooperative-Intelligent Transportation System)는 전방 충돌 경고, 도로 안전 서비스 및 비상 정지와 같은 엄격한 실시간성이 요구되는 서비스들을 제공한다. C-ITS의 핵심 기술인 WAVE(Wireless Access in Vehicular Environments)는 고속 이동 환경을 위해 고안된 기술이지만, 실제 도로 환경에 적용하여 안정적인 통신 서비스를 제공하기 위해서는 실차 환경의 다양한 성능 시험이 요구된다. 실제 도로 환경에서 WAVE 통신 성능은 이동 중인 차량, 도로의 형상 및 지형 등의 주변 환경에 의해 영향을 받으며 특히, 차량이 고속으로 이동하는 경우 차량의 속도에 따라 주행하는 위치와 노변장치와의 접근성 등 주변 환경이 빠르게 변화한다. 이 변화는 통신 성능에 영향을 주는 요소이므로 이를 분석하기 위한 시스템과 방법이 필요하다. 본 논문에서는 고속 주행 환경에서 효과적인 성능 시험 시스템 구성과 시험 방법을 제안하고, 실차 시험을 통해 측정한 데이터를 기반으로 통신 성능을 분석한 결과를 제시한다.

• 한국ITS학회 논문지
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• 제18권6호
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• pp.176-190
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• 2019

최근 횡단보도 및 교차로에서 보행자 충돌사고 예방의 중요성이 증가하고 있다. C-ITS 서비스에서 이러한 사고를 줄이기 위하여 보행자 충돌방지 경고 서비스를 제시하고 있다. 그러나 현재 C-ITS 표준에 따른 보행자 충돌방지 경고 서비스는 현장의 카메라에서 보행자를 바로 검출하여 서비스를 제공하는 것이 아니라 관제 센터의 영상분석 서버에서 보행자를 검출하고 ITS 시스템과 연계하여 서비스를 제공하기 때문에 실시간성을 만족하기 어렵다. 본 논문에서는 엣지 카메라를 이용하여 현장에서 보행자를 검출하고 검출결과를 V2X 인프라를 통해 바로 운전자에게 제공하는 보행자 충돌방지 경고 시스템을 제안하고, 구현한 후 성능 평가를 시행하였다. 평가 결과, 최악의 상황에서도 보행자 충돌방지 경고 메시지를 C-ITS 표준에서 요구하고 있는 300ms 이내의 지연시간을 만족하여 전달할 수 있음을 확인하였다.

• Journal of Advanced Marine Engineering and Technology
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• 제37권8호
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• pp.918-931
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• 2013

기존의 Pan-Tilt Zooming 카메라를 이용한 객체 추적은 카메라를 통해 획득한 영상을 분석하여 물체의 이동을 검출하였다. 그러나 이 방식은 영상처리를 위한 알고리즘과 이를 위한 하드웨어 비용이 소요되었다. 그리고 주변 환경의 변화나 영상 잡음 등으로 유효하지 않은 영상이 획득되는 경우나 사각지대가 존재하여 영상의 획득 자체가 불가능한 경우에는 대응이 어렵다. 본 논문에서는 영상 정보처리 기반의 객체 추적 기술이 가지는 한계를 극복하기 위해 무선 센서 네트워크 기반의 IEEE 802.15.4 표준을 적용한 GPS수신기가 장착된 센서노드를 시설물에 장착하여 센서노드의 가속도 센서가 장착된 시설물의 움직임을 인식하고 해당 센서노드의 GPS 정보를 이용하여 물체를 추적할 수 있는 시스템을 제안한다. 이 시스템은 영상처리에 요구되는 연산량을 감소시킴과 동시에 넓은 지역의 이동 물체를 추적할 수 있도록 효율적인 PTZ 카메라 제어를 가능하게 한다. 또한 센서노드의 유동적 장착이 가능할 수 있도록 무선의 메쉬 네트워크를 구현하여 설치의 효율성을 올렸다.