• Journal of Microbiology and Biotechnology
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• v.24 no.7
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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.

• Progress in Medical Physics
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• v.29 no.1
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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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• v.22 no.5
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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.

• Progress in Medical Physics
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• v.28 no.4
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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.

• Progress in Medical Physics
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• v.30 no.4
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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.

• Proceedings of the Korean Magnestics Society Conference
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• 2015.05a
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• pp.59-59
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• 2015

• Korean Journal of Radiology
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• v.21 no.5
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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.

• The Journal of Korean Institute of Communications and Information Sciences
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• v.42 no.5
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• pp.1069-1076
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• 2017

C-ITS(Cooperative-Intelligent Transportation System) provides services that require strict real-time such as forward collision warning, road safety service and emergency stop. WAVE(Wireless Access in Vehicular Environments), a core technology of C-ITS, is a technology designed for high-speed driving. However, in order to provide stable communication service by applying to real road environment, various performance tests of real vehicular environment are required. In the real road environment, WAVE communication performance is influenced by the surrounding environment such as moving vehicle, road shape and topography. Especially, when the vehicle is moving at high speed, the traveling position according to the speed of the vehicle, The surrounding environment changes rapidly. Such changes are factors affecting the communication performance, therefore a system and methods for analyzing them are needed. In this paper, we propose the configuration and test method of an effective performance evaluation system under high-speed driving and describe the results of analyzing the communication performance based on the data measured through the actual vehicle test.

• The Journal of The Korea Institute of Intelligent Transport Systems
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• v.18 no.6
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• pp.176-190
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• 2019

The prevention of pedestrian accidents in crosswalks and intersections is very important. The C-ITS services provide a warning service for preventing accidents between cars and pedestrians. In the current pedestrian collision prevention warning service according to the C-ITS standard, however, it is difficult to provide real-time service because it detects pedestrians from a video-analysis server in the control center and sends service messages through the ITS system. This paper proposes a pedestrian collision-prevention warning system that detects pedestrians in the local field using an edge camera and sends a warning message directly to the driver through a roadside unit. An evaluation showed that the proposed system could deliver the pedestrian collision prevention-warning message to the driver satisfying the delay time within the 300 ms required by the C-ITS standard, even in the worst case.

• Journal of Advanced Marine Engineering and Technology
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• v.37 no.8
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• pp.918-931
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• 2013

The conventional object tracking techniques using PTZ camera detects object movements by analyzing acquired image. However, this technique requires expensive hardware devices to perform a complex image processing. And it is occasionally hard to detect object movements, if an acquired image is low quality or image acquisition is impossible. In this paper, we proposes a smart security CCTV system applying to wireless sensor network technique based on IEEE 802.15.4 standard to overcome the problems of conventional object tracking technique, which enables to track suspicious objects by detecting object movements and GPS data in sensor node. This system enables an efficient control of PTZ camera to observe a wide area, decreasing image processing complexity. Also, wireless sensor network is implemented using mesh networks to increase the efficiency of installing sensor node.