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Amino Acid Biosynthesis and Gene Regulation in Seed (종자내 아미노산 합성 조절 유전자에 관한 연구)

  • ;;;;;Fumio Takaiwa
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.61-74
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    • 1996
  • Human and monogastric animals can not synthesize 10 out of the 20 amino asids and therefor need to obtain these from their diet. The plant seed is a major source of dietary protein. It is particular important in their study to increase nutritional quality of the seed storage proteins. The low contents of lysine, asparagine and threonenein various cereal seeds and of cystein and methionine. In legume seeds is due to the low proportions of these amino acids in the major storage proteins, we have tried to apply the three strategies; (1) mutagenesis and selection of specific amino acid analogue resistance, (2) cloning and expression study of lysine biosynthesis related gene, (3) transfomation of lysine rich soybean glycinin gene. The 5-methyltryptophan (5MT) resistant cell lines, SAR1, SAR2 and SAR3 were selected from anther derived callus of rice (Oryza sativa L. "Sasanishiki"). Among these selected cell lines, two (SAR1 and SAR3) were able to grow stably at 200 mg/L of 5MT. Analysis of the freed amino acids in callus shows that 5MT resistant cells (SAR3) accumulated free tryptophan at least up to 50 times higher than those that of the higher than of SAS. These results indicated that the 5MT resistant cell lines are useful in studies of amino acid biosynthesis. Tr75, a rice (Oryza sativa L., var. Sasanishiki) mutant resistant to 5MT was segregated from the progenies of its initial mutant line, TR1. The 5MT resistant of TR75 was inherited in the M8 generations as a single dominant nuclear gene. The content of free amino acids in the TR75 homozygous seeds increased approximately 1.5 to 2.0 fold compared to wild-type seeds. Especially, the contents of tryptophan, phenylalanine and aspartic acid were 5.0, 5.3 and 2.7 times higher than those of wild-type seeds, respectively. The content of lysine is significantly low in rice. The lysine is synthesized by a complex pathway that is predominantly regulated by feedback inhibition of several enzymes including asparginase, aspatate kinase, dihydrodipicolinat synthase, etc. For understanding the regulation mechanism of lysine synthesis in rice, we try to clone the lysine biosynthetic metabolism related gene, DHPS and asparaginase, from rice. We have isolated a rice DHPS genomic clone which contains an ORF of 1044 nucleotides (347 amino acids, Mr. 38, 381 daltons), an intron of 587 nucleotides and 5'and 3'-flanking regions by screening of rice genomic DNA library. Deduced amino acid sequence of mature peptide domain of GDHPS clone is highly conserved in monocot and dicot plants whereas that of transit peptide domain is extremely different depending on plant specie. Southern blot analysis indicated that GDHPS is located two copy gene in rice genome. The transcripts of a rice GDHPS were expressed in leaves and roots but not detected in callus tissues. The transcription level of GDHPS is much higher in leaves indicating enormous chloroplast development than roots. Genomic DNA clones for asparaginase genes were screened from the rice genomic library by using plaque hybridization technique. Twelve different genomic clones were isolated from first and second screening, and 8 of 12 clones were analyzed by restriction patterns and identified by Southern Blotting, Restriction enzyme digestion patterns and Southern blot analysis of 8 clones show the different pattern for asparaginase gene. Genomic Southern blot analysis from rice were done. It is estimated that rice has at least 2-3 copy of asparaginase gene. One of 8 positive clones was subcloned into the pBluescript SK(+) vector, and was constructed the physical map. For transformation of lysine rich storage protein into tobacco, soybean glycinin genes are transformed into tobacco. To examine whether glycinin could be stably accumulated in endosperm tissue, the glycinin cDNA was transcriptionally fused to an endosperm-specific promotor of the rice storage protein glutelin gene and then introduced into tobacco genomic via Agrobacterium-mediated transformation. Consequently the glycinin gene was expressed in a seed-and developmentally-specific manner in transgenic tobacco seeds. Glycinin were targeted to vacuole-derived protein bodies in the endosperm tissue and highly accumulated in the matrix region of many transgenic plant (1-4% of total seed proteins). Synthesized glycinin was processed into mature form, and assembled into a hexamer in a similar manner as the glycinin in soybean seed. Modified glycinin, in which 4 contiguous methionine residues were inserted at the variable regions corresponding to the C - teminal regions of the acidic and basic polypeptides, were also found to be accumulated similarly as in the normal glycinin. There was no apparent difference in the expression level, processing and targeting to protein bodies, or accumulation level between normal and modified glycinin. glycinin.

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Experimental Validation of Isogeometric Optimal Design (아이소-지오메트릭 형상 최적설계의 실험적 검증)

  • Choi, Myung-Jin;Yoon, Min-Ho;Cho, Seonho
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.27 no.5
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    • pp.345-352
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    • 2014
  • In this paper, the CAD data for the optimal shape design obtained by isogeometric shape optimization is directly used to fabricate the specimen by using 3D printer for the experimental validation. In a conventional finite element method, the geometric approximation inherent in the mesh leads to the accuracy issue in response analysis and design sensitivity analysis. Furthermore, in the finite element based shape optimization, subsequent communication with CAD description is required in the design optimization process, which results in the loss of optimal design information during the communication. Isogeometric analysis method employs the same NURBS basis functions and control points used in CAD systems, which enables to use exact geometrical properties like normal vector and curvature information in the response analysis and design sensitivity analysis procedure. Also, it vastly simplify the design modification of complex geometries without communicating with the CAD description of geometry during design optimization process. Therefore, the information of optimal design and material volume is exactly reflected to fabricate the specimen for experimental validation. Through the design optimization examples of elasticity problem, it is experimentally shown that the optimal design has higher stiffness than the initial design. Also, the experimental results match very well with the numerical results. Using a non-contact optical 3D deformation measuring system for strain distribution, it is shown that the stress concentration is significantly alleviated in the optimal design compared with the initial design.

Application of CSP Filter to Differentiate EEG Output with Variation of Muscle Activity in the Left and Right Arms (좌우 양팔의 근육 활성도 변화에 따른 EEG 출력 구분을 위한 CSP 필터의 적용)

  • Kang, Byung-Jun;Jeon, Bu-Il;Cho, Hyun-Chan
    • Journal of IKEEE
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    • v.24 no.2
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    • pp.654-660
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    • 2020
  • Through the output of brain waves during muscle operation, this paper checks whether it is possible to find characteristic vectors of brain waves that are capable of dividing left and right movements by extracting brain waves in specific areas of muscle signal output that include the motion of the left and right muscles or the will of the user within EEG signals, where uncertainties exist considerably. A typical surface EMG and noninvasive brain wave extraction method does not exist to distinguish whether the signal is a motion through the degree of ionization by internal neurotransmitter and the magnitude of electrical conductivity. In the case of joint and motor control through normal robot control systems or electrical signals, signals that can be controlled by the transmission and feedback control of specific signals can be identified. However, the human body lacks evidence to find the exact protocols between the brain and the muscles. Therefore, in this paper, efficiency is verified by utilizing the results of application of CSP (Common Spatial Pattern) filter to verify that the left-hand and right-hand signals can be extracted through brainwave analysis when the subject's behavior is performed. In addition, we propose ways to obtain data through experimental design for verification, to verify the change in results with or without filter application, and to increase the accuracy of the classification.

A study on the connected-digit recognition using MLP-VQ and Weighted DHMM (MLP-VQ와 가중 DHMM을 이용한 연결 숫자음 인식에 관한 연구)

  • Chung, Kwang-Woo;Hong, Kwang-Seok
    • Journal of the Korean Institute of Telematics and Electronics S
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    • v.35S no.8
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    • pp.96-105
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    • 1998
  • The aim of this paper is to propose the method of WDHMM(Weighted DHMM), using the MLP-VQ for the improvement of speaker-independent connect-digit recognition system. MLP neural-network output distribution shows a probability distribution that presents the degree of similarity between each pattern by the non-linear mapping among the input patterns and learning patterns. MLP-VQ is proposed in this paper. It generates codewords by using the output node index which can reach the highest level within MLP neural-network output distribution. Different from the old VQ, the true characteristics of this new MLP-VQ lie in that the degree of similarity between present input patterns and each learned class pattern could be reflected for the recognition model. WDHMM is also proposed. It can use the MLP neural-network output distribution as the way of weighing the symbol generation probability of DHMMs. This newly-suggested method could shorten the time of HMM parameter estimation and recognition. The reason is that it is not necessary to regard symbol generation probability as multi-dimensional normal distribution, as opposed to the old SCHMM. This could also improve the recognition ability by 14.7% higher than DHMM, owing to the increase of small caculation amount. Because it can reflect phone class relations to the recognition model. The result of my research shows that speaker-independent connected-digit recognition, using MLP-VQ and WDHMM, is 84.22%.

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Study on CsRCI2D and CsRCI2H for improvement of abiotic stress tolerance in Camelina sativa L.

  • Lim, Hyun-Gyu;Kim, Hyun-Sung;Kim, Jung-Eun;Ahn, Sung-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.196-196
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    • 2017
  • Oilseed crop Camelina (Camelina sativa L.) is a suitable for biodiesel production that has high adaptability under low-nutrient condition like marginal land and requires low-input cost for cultivation. Enhanced abiotic stress tolerance of Camelina is very important for oil production under the wide range of different climate. CsRCI2s (Rare Cold Inducible 2) are related proteins in various abiotic stresses that predicted to localized at plasma membrane (PM) and endoplasmic reticulum (ER). These proteins are consist of eight-family that can be divided into tail (CsRCI2D/E/F/G) and no-tail (CsRCI2A/B/E/H) type of C-terminal. However, it is still less understood the function of C-terminal tail. In this study, CsRCI2D/H genes were cloned through gateway cloning system that used pCB302-3 as destination vector. And we used agrobacterium-mediated transformation system for generation of overexpression (OX) transformants. Overexpression of target gene was confirmed using RT-PCR and segregation ratio on selection media. We analyzed physiological response in media and soil under abiotic stresses using CsRCI2D and CsRCI2H overexpression plant. To compare abiotic stresses tolerance, wild type and CsRCI2D/H OX line seeds were sown on agar plate treated with various NaCl and mannitol concentration for 7 days. In the test of growth rate under abiotic stress on media, CsRCI2H OX line showed similar to NaCl and mannitol stress. In the other hand, CsRCI2D OX line showed to be improved stress tolerance that especially increased in 200mM NaCl but was similar on mannitol media. In greenhouse, WT and CsRCI2D/H OX lines for physiological analysis and productivity under abiotic stresses were treated 100, 150, 200mM NaCl. Then it was measured various parameters such as leaf width and length, plant height, total seed weight, flower number, seed number. CsRCI2H OX line in greenhouse did not show any changes in physiological parameters but CsRCI2D OX line was improved both physiological response and productivity under NaCl stress. Among physiological parameters of CsRCI2D OX line under NaCl stress, leaf length and width were observed shorter than WT but it were slightly longer than WT in 200mM NaCl stress. Furthermore, total seed weight of CsRCI2D OX line under stress displayed to decrease than WT in normal condition, but it was gradually raised with increasing NaCl stress then more than WT relatively. These results suggested CsRCI2D might be contribute to improve abiotic stress tolerance. However, function of CsRCI2H is need to more detail study. In conclusion, overexpression of CsRCI2s family can generate various environmental stress tolerance plant and may improve crop productivity for bio-energy production.

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TLS (Total Least-Squares) within Gauss-Helmert Model: 3D Planar Fitting and Helmert Transformation of Geodetic Reference Frames (가우스-헬머트 모델 전최소제곱: 평면방정식과 측지좌표계 변환)

  • Bae, Tae-Suk;Hong, Chang-Ki;Lim, Soo-Hyeon
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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    • v.40 no.4
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    • pp.315-324
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    • 2022
  • The conventional LESS (LEast-Squares Solution) is calculated under the assumption that there is no errors in independent variables. However, the coordinates of a point, either from traditional ground surveying such as slant distances, horizontal and/or vertical angles, or GNSS (Global Navigation Satellite System) positioning, cannot be determined independently (and the components are correlated each other). Therefore, the TLS (Total Least Squares) adjustment should be applied for all applications related to the coordinates. Many approaches were suggested in order to solve this problem, resulting in equivalent solutions except some restrictions. In this study, we calculated the normal vector of the 3D plane determined by the trace of the VLBI targets based on TLS within GHM (Gauss-Helmert Model). Another numerical test was conducted for the estimation of the Helmert transformation parameters. Since the errors in the horizontal components are very small compared to the radius of the circle, the final estimates are almost identical. However, the estimated variance components are significantly reduced as well as show a different characteristic depending on the target location. The Helmert transformation parameters are estimated more precisely compared to the conventional LESS case. Furthermore, the residuals can be predicted on both reference frames with much smaller magnitude (in absolute sense).

Development of an Anomaly Detection Algorithm for Verification of Radionuclide Analysis Based on Artificial Intelligence in Radioactive Wastes (방사성폐기물 핵종분석 검증용 이상 탐지를 위한 인공지능 기반 알고리즘 개발)

  • Seungsoo Jang;Jang Hee Lee;Young-su Kim;Jiseok Kim;Jeen-hyeng Kwon;Song Hyun Kim
    • Journal of Radiation Industry
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    • v.17 no.1
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    • pp.19-32
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    • 2023
  • The amount of radioactive waste is expected to dramatically increase with decommissioning of nuclear power plants such as Kori-1, the first nuclear power plant in South Korea. Accurate nuclide analysis is necessary to manage the radioactive wastes safely, but research on verification of radionuclide analysis has yet to be well established. This study aimed to develop the technology that can verify the results of radionuclide analysis based on artificial intelligence. In this study, we propose an anomaly detection algorithm for inspecting the analysis error of radionuclide. We used the data from 'Updated Scaling Factors in Low-Level Radwaste' (NP-5077) published by EPRI (Electric Power Research Institute), and resampling was performed using SMOTE (Synthetic Minority Oversampling Technique) algorithm to augment data. 149,676 augmented data with SMOTE algorithm was used to train the artificial neural networks (classification and anomaly detection networks). 324 NP-5077 report data verified the performance of networks. The anomaly detection algorithm of radionuclide analysis was divided into two modules that detect a case where radioactive waste was incorrectly classified or discriminate an abnormal data such as loss of data or incorrectly written data. The classification network was constructed using the fully connected layer, and the anomaly detection network was composed of the encoder and decoder. The latter was operated by loading the latent vector from the end layer of the classification network. This study conducted exploratory data analysis (i.e., statistics, histogram, correlation, covariance, PCA, k-mean clustering, DBSCAN). As a result of analyzing the data, it is complicated to distinguish the type of radioactive waste because data distribution overlapped each other. In spite of these complexities, our algorithm based on deep learning can distinguish abnormal data from normal data. Radionuclide analysis was verified using our anomaly detection algorithm, and meaningful results were obtained.

On-line Image Guided Radiation Therapy using Cone-Beam CT (CBCT) (콘빔CT (CBCT)를 이용한 온라인 영상유도방사선치료 (On-line Image Guided Radiation Therapy))

  • Bak, Jin-O;Jeong, Kyoung-Keun;Keum, Ki-Chang;Park, Suk-Won
    • Radiation Oncology Journal
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    • v.24 no.4
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    • pp.294-299
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    • 2006
  • $\underline{Purpose}$: Using cone beam CT, we can compare the position of the patients at the simulation and the treatment. In on-line image guided radiation therapy, one can utilize this compared data and correct the patient position before treatments. Using cone beam CT, we investigated the errors induced by setting up the patients when use only the markings on the patients' skin. $\underline{Materials\;and\;Methods}$: We obtained the data of three patients that received radiation therapy at the Department of Radiation Oncology in Chung-Ang University during August 2006 and October 2006. Just as normal radiation therapy, patients were aligned on the treatment couch after the simulation and treatment planning. Patients were aligned with lasers according to the marking on the skin that were marked at the simulation time and then cone beam CTs were obtained. Cone beam CTs were fused and compared with simulation CTs and the displacement vectors were calculated. Treatment couches were adjusted according to the displacement vector before treatments. After the treatment, positions were verified with kV X-ray (OBI system). $\underline{Results}$: In the case of head and neck patients, the average sizes of the setup error vectors, given by the cone beam CT, were 0.19 cm for the patient A and 0.18 cm for the patient B. The standard deviations were 0.15 cm and 0.21 cm, each. On the other hand, in the case of the pelvis patient, the average and the standard deviation were 0.37 cm and 0.1 cm. $\underline{Conclusion}$: Through the on-line IGRT using cone beam CT, we could correct the setup errors that could occur in the conventional radiotherapy. The importance of the on-line IGRT should be emphasized in the case of 3D conformal therapy and intensity-modulated radiotherapy, which have complex target shapes and steep dose gradients.

Isogeometric Shape Design Optimization of Power Flow Problems at High Frequencies (고주파수 파워흐름 문제의 아이소-지오메트릭 형상 최적설계)

  • Yoon, Minho;Ha, Seung-Hyun;Cho, Seonho
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.27 no.3
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    • pp.155-162
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    • 2014
  • Using an isogeometric approach, a continuum-based shape design optimization method is developed for steady state power flow problems at high frequencies. In case the isogeometric method is employed to the shape design optimization, the NURBS basis functions used in CAD geometric modeling are directly utilized to embed the exact geometry into the computational framework so that the design parameterization for shape optimization is much easier than that in the finite element method and consequently provides the enhanced smoothness of design perturbations. Thus, exact geometric models can be used in both the response and the shape sensitivity analyses, where normal vector and curvature are continuous over the whole design space so that enhanced shape sensitivity can be expected. Through numerical examples, the developed isogeometric sensitivity is compared with finite difference one to provide excellent agreement. Also, it turns out that the proposed method works very well in the shape optimization problems.

EZH2-Mediated microRNA-139-5p Regulates Epithelial-Mesenchymal Transition and Lymph Node Metastasis of Pancreatic Cancer

  • Ma, Jin;Zhang, Jun;Weng, Yuan-Chi;Wang, Jian-Cheng
    • Molecules and Cells
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    • v.41 no.9
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    • pp.868-880
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    • 2018
  • Pancreatic cancer (PC) is one of the most aggressive cancers presenting with high rates of invasion and metastasis, and unfavorable prognoses. The current study aims to investigate whether EZH2/miR-139-5p axis affects epithelial-mesenchymal transition (EMT) and lymph node metastasis (LNM) in PC, and the mechanism how EZH2 regulates miR-139-5p. Human PC and adjacent normal tissues were collected to determine expression of EZH2 and miR-139-5p, and their relationship with clinicopathological features of PC. Human PC cell line was selected, and treated with miR-139-5p mimics/inhibitors, EZH2 vector or shEZH2 in order to validate the regulation of EZH2-mediated miR-139-5p in PC cells. Dual-luciferase report gene assay and chromatin immunoprecipitation assay were employed to identify the relationship between miR-139-5p and EZH2. RT-qPCR and Western blot analysis were conducted to determine the expression of miR-139-5p, EZH2 and EMT-related markers and ZEB1/2. Tumor formation ability and in vitro cell activity were also analyzed. Highly-expressed EZH2 and poorly-expressed miR-139-5p were detected in PC tissues, and miR-139-5p and EZH2 expressions were associated with patients at Stage III/IV, with LNM and highly-differentiated tumors. EZH2 suppressed the expression of miR-139-5p through up-regulating Histone 3 Lysine 27 Trimethylation (H3K27me3). EMT, cell proliferation, migration and invasion were impeded, and tumor formation and LNM were reduced in PC cells transfected with miR-139-5p mimics and shEZH2. MiR-139-5p transcription is inhibited by EZH2 through up-regulating H3K27me3, thereby down-regulation of EZH2 and up-regulation of miR-139-5p impede EMT and LNM in PC. In addition, the EZH2/miR-139-5p axis presents as a promising therapeutic strategy for the treatment of PC.