• Journal of Applied Biological Chemistry
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• 제44권4호
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• pp.167-172
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• 2001

A cDNA fragment encoding the chloroplastic fructose-1, 6-bisphosphatase (FBPase) was cloned via PCR from the cDNA library of pea leaves. The cloned cDNA, about 1.05 kbp without signal sequence, was introduced into a pET-28a vector for expression in E. coli strain BL21(DE3)pLysS. The recombinant FBPase was purified through $Ni^+-NTA$ affinity chromatography and characterized. Molecular mass of the monomer was about 42,000. Enzymatic activity of the purified enzyme as the native pea chloroplastic FBPase was the highest at alkaline pH (pH 9.0). The recombinant enzyme was activated by a reducing agent DTT and was insensitive to AMP. The activation energy (Ea) and Arrehenius frequency factor were 42.67 kcal/mol and $2.65{\times}10^{14}/s$, respectively, slightly higher than those of the native enzyme. $K_M$ and $V_{max}$ were $99.98{\mu}M$ and $52.9{\mu}M/min$, respectively, which were comparable with the native enzyme.

• 천문학회보
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• 제42권2호
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• pp.60.2-61
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• 2017

A Convolutional Neural Network(CNN) is one of the well-known deep-learning methods in image processing and computer vision area. In this study, we apply CNN to two kinds of flare forecasting models: flare classification and occurrence. For this, we consider several pre-trained models (e.g., AlexNet, GoogLeNet, and ResNet) and customize them by changing several options such as the number of layers, activation function, and optimizer. Our inputs are the same number of SOHO)/MDI images for each flare class (None, C, M and X) at 00:00 UT from Jan 1996 to Dec 2010 (total 1600 images). Outputs are the results of daily flare forecasting for flare class and occurrence. We build, train, and test the models on TensorFlow, which is well-known machine learning software library developed by Google. Our major results from this study are as follows. First, most of the models have accuracies more than 0.7. Second, ResNet developed by Microsoft has the best accuracies : 0.77 for flare classification and 0.83 for flare occurrence. Third, the accuracies of these models vary greatly with changing parameters. We discuss several possibilities to improve the models.

• 한국멀티미디어학회논문지
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• 제25권9호
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• pp.1257-1265
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• 2022

Recognition of sign language is challenging due to the occlusion of hands, accuracy of hand gestures, and high computational costs. In recent years, deep learning techniques have made significant advances in this field. Although these methods are larger and more complex, they cannot manage long-term sequential data and lack the ability to capture useful information through efficient information processing with faster convergence. In order to overcome these challenges, we propose a word-level sign language recognition (SLR) system that combines a real-time human pose detection library with the minimized version of the gated recurrent unit (GRU) model. Each gate unit is optimized by discarding the depth-weighted reset gate in GRU cells and considering only current input. Furthermore, we use sigmoid rather than hyperbolic tangent activation in standard GRUs due to performance loss associated with the former in deeper networks. Experimental results demonstrate that our pose-based optimized GRU (Pose-OGRU) outperforms the standard GRU model in terms of prediction accuracy, convergency, and information processing capability.

• Journal of Microbiology
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• 제37권3호
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• pp.154-158
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• 1999

The arginine residue at position 243 (Arg 243) of the yeast transcription factor, Gcn4p, is invariably conserved among bZIP transcription factors. Using site-directed oligonucleotide saturation mutagenesis involving two-step polymerase chain reaction (PCR) amplification, random mutations were successfully introduced at the codon of 243 in the basic domain of Gcn4p. This mutant library was transformed ito Gcn4p defective yeast strain and selected for the transcriptionally active colonies. All colonies which were transcriptionally active had arginines in the codon 243. In this study, the strand preference by Taq polymerase during mutagenesis was also tested. Oligonucleotides were specially designed to test whether or not the polymerase was preferred using the strand as a template. A population of randomly mutated products were cloned into an appropriate vector and characterized by DNA sequencing analysis. Saturation mutagenesis which was performed efficiently by this method revealed a strong bias in terms of strand preference of Taq polymerase by an approximate ratio of 3 to 1 in this study.

• Journal of Microbiology and Biotechnology
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• 제22권5호
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• pp.600-606
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• 2012

Studies into the cell death program termed apoptosis have resulted in new information regarding how cells control and execute their own demise, including insights into the mechanism by which death-preventing factors can inhibit Bax-induced caspase activation. We investigated high temperature stress-induced cell death in Brassica rapa. Using a yeast functional screening from a Brassica rapa cDNA library, the BH5-127 EST clone encoding an apoptotic suppressor peptide was identified. However, a phylogenic tree showed that BH5-127 clusters within a clade containing SUMO-1 (Small Ubiquitin-like Modifier-1). BH5-127 was confirmed similar to have function to SUMO-1 as Fas suppression. Expression of BH5-127 showed that substantial suppression of cell death survived on SD-galactose-$Leu^-$-$Ura^-$ medium. The results suggest that BrSE ($\underline{B}$rassica rapa $\underline{S}$entrin $\underline{E}$ST, BH5-127) is one of the important regulatory proteins in programming cell death, especially in the seedling stage of Chinese cabbage.

• Molecules and Cells
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• 제23권3호
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• pp.316-322
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• 2007

Zinc finger transcription factor genes are a significant fraction of the genes in the vertebrate genome. Here we report the isolation and characterization of a human zinc finger-containing gene, ZNF435, from a fetal brain cDNA library. ZNF435 cDNA is 1290 base pairs in length and contains an open reading frame encoding 349 amino acids with four C2H2-type zinc fingers at its carboxyl terminus and a SCAN motif at its amino terminus. RT-PCR results showed that ZNF435 was expressed in all tested tissues. A ZNF435-GFP fusion protein was located in the nucleus and the four zinc fingers acted as nuclear localization signals (NLSs). ZNF435 was found to be capable of homo-association, and this effect was independent of its zinc fingers. Furthermore, ZNF435 proved to be a transcription repressor as its overexpression in AD293 cells inhibited the transcriptional activities of AP-1.

• BMB Reports
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• 제51권5호
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• pp.236-241
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• 2018

Anoctamin 1 (ANO1) is an anion channel that is activated by changes in cytosolic $Ca^{2+}$ concentration and noxious heat. Although the critical roles of ANO1 have been elucidated in various cell types, the control of its gating mechanisms by $Ca^{2+}$ and heat remain more elusive. To investigate critical amino acid residues for modulation of $Ca^{2+}$ and heat sensing, we constructed a randomized mutant library for ANO1. Among 695 random mutants, reduced $Ca^{2+}$ sensitivity was observed in two mutants (mutant 84 and 87). Consequently, the E143A mutant showed reduced sensitivity to $Ca^{2+}$ but not to high temperatures, whereas the E705V mutant exhibited reduced sensitivity to both $Ca^{2+}$ and noxious heat. These results suggest that the glutamic acids (E) at 143 and 705 residues in ANO1 are critical for modulation of $Ca^{2+}$ and/or heat responses. Furthermore, these findings help to provide a better understanding of the $Ca^{2+}$-mediated activation and heat-sensing mechanism of ANO1.

• BMB Reports
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• 제39권6호
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• pp.709-716
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• 2006

Mammalian polo-like kinase 1 (Plk1) acts at various stages in early and late mitosis. Plk1 localizes at the centrosome and maintains this position through mitosis. Thereafter Plk1 moves to the kinetochore and midbody region, important sites during chromosome separation and cytokinesis. The catalytic domain of Plk1 is in the N-terminus region, whereas the non-catalytic region in the C-terminus of Plk1 has a conserved motif, named the Polobox. This motif is critical for Plk localization. EGFP proteins fused with the N-terminus and C-terminus of Plk1 localize in the nucleus and centrosomes, respectively. The core sequences of the polo-box (50 amino acids) also localize in Plk1 target organelles. To screen for domain-specific target proteins of Plk1, we constructed an N-terminal domain and a tandem repeat polo-box motif, and used them as templates in a yeast two-hybrid screen. The HeLa cell cDNA library indicated several proteins including the centrosome/kinetochore components or regulators, to be characterized as positive clones. Through in vitro protein binding analyses, we confirmed an interaction between these proteins and Plk1. The data reported from this study indicate that the N- and C- termini of Plk1 may function through recruitment and/or activation of domain-specific target proteins in dividing cells. Additionally, tandem repeats of the conserved core motif of the polo-box are sufficient for targeting and may be useful as a centrosome/kinetochore-specific targeting peptide.

• 대한간호학회지
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• 제8권1호
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• pp.16-42
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• 1978

This survey was made for a month starting from November 15 to December 15, 1977 covering 711 students taking the junior. (3rd-yea.) and the senior. (4th-year) at nine college of nursing in Seoul concerning their perception and Attitude toward the profession of nursing, motive for the necessity of learning, environment of study, attitude of study and particulars relevant with study performance, particulars of library, references and reading, assignments and particulars of the degree of confidence for the learning achievement. Through the survey of the above Particulars, the following results were obtained by classifying all subject matters and by analysing motive of the selection of their course, awarding or not awarding of scholarships. 1. General characteristics: it was revealed that 406 students (57.1%) were attending at the junior. while 305 students (42.9%) were taking the senior. Thus, the total number was 711 and their average age was 21.4 years. Their dwelling category was; 73.9 percent of them resided at their parent's home, 214 students (30.1%) were awarded with scholarships. The reason to be attracted by nursing science was the possibility of continuing social life after graduation (43.5%). 2. Their perception and attitude toward the profession of nursing: According to the perception of profession by the students of each grade, students of the 4th grade showed comparatively strong conception. Also, students of the 4th grade showed more positive attitude in the purchase and reading of magazines relative with the science of nursing, in the reading of Code for Nurses and in their interest in the activity of nursing field. For the necessity of mission of nurse, 97.7 percent of the entire number of students covered responded to necessity. For the necessity of the particular humanity and particularity in the character of nurses, 95.8 percent of those students responded to necessity. By the each grade, students of the 4th grade showed more response. 3. As to professional field desired after completing the professional course: 57 percent of those students desired for clinician nurse while 55 percents desired for community health-nurse. 4. As to the environment of study: they were mostly satisfied with their present residential environment. However, they complained of inconvenience at their lecture-halls. Students of the 3rd grade showed more complain. As to their attitude toward the adjustment of environment of study, they showed a affirmative response. As to the opinion of factors which interfere with their study, comparatively strong response was showed in their scepticism in the science of nursing, insufficient comprehension in general learning, relation with professors n4 discrepancy in the method of study. According to opinions of students at each grade, students of the 4th grade showed more scepticism. 5. Particulars relative with their attitude and performance of study : As to their knowledge of the objectives of their study of subject, the majority was to study with a partial knowledge of the objectives of their study. As to the plan of study, a low percentage indicated management of routine life under regular scheduling. Students of the 4th grade responded to rather planned life. As to time spent in independent study, response to concentrated study when necessary was stronger than that to regular daily study. Students of the 4th grade showed stronger response to regular study than that of the 3rd grade. As to the contents of their note-taking, 67.4 percents of those students responded to such regulatory procedure performing in the lecture-hall as they listen to lectures. 17.3 percents of those students showed response to adding supplementary informations from references to what was entered in choir note-taking at their lecture-halls. 6. Particulars of library, references and reading books: As to receiving of instruction for the utilization of library and time of receiving such instruction 64.7 percents of those students had received such instruction. 66.7 percents of the those responded received such instruction at orientation conducted for freshmen. As to the convenience of the utilization of library, 49.9 percents of those students responded to convenience. However, students of the 3rd grade showed a much stronger response to inconvenience. As to the time of the utilization of library,92.5 percents of those students showed a response to occasional utilization for particular purpose than regular utilization. 53.2 percents of those students responded to ordinary in quantity that library have references. 34.2 percents of those students responded to insufficient. As to the particular relative with the method and field of reading: 53.5 percents of those students responded to intensive reading and was the majority. As to the reading field, fiction u as the majority. When read any books for their major, they usually rend Korean text-b, oks. 7. Particular relative with giving assignment: All respondents were well aware of the objectives of giving home tasks. As to the attitude toward assignments and performing home tasks, 54.8 percents of those students to making ostentatious study because of an excessive quantity of assignments imposed. For performing assignment, they showed comparatively positive response. Also, 52.2 percents of those students responded that they usually submitted complected assignment with references. 8. As to motive to realize the necessity of study : 55.6 percents of those students responded that they realized such necessity in communication with patients when they were engaged in clinical practice. Also, 8.6, the lowest percents of those students responded that they realized such necessity in the course of conversation with nurses when they were engaged in clinical practice. 9. As to the determination of their confidence in the performance of study relative with clinical experience: They showed a general inclination of having in nursing. The major response was that they came to well comprehend the patients families. the lowest response was that they could apply what was learned at lecture-hall to practice. This response incidentally showed the distance the lecture-hall and practical study. In general items, students of the 4th grade showed more favorable response than students of the 3rd grade and there was a significant difference. 10. As to the perception and attitude toward profession according to the motive of selecting the nursing science : Those who selected the nursing voluntarily showed stronger conception than those who selected the nursing through indirect influence. However, there was no significant difference on this point. Only there was a remarkable difference in the reading of Code for Nurses. 11. Those who showed a stronger conception in the profession of nursing according to the motives of attractive nursing science indicated a strong will and ability to manage stable life and comparatively strong response was shown in the management of good home life because of the good adaptability of the science to their character. This group showed a strong conception of the profession than those who responded that they prefer this profession out of a longing for the work of a hospital and for the easy obtaining of opportunity to immigrate to over seas and for economic cause and for high school grade. There was significant difference between these two groups, 12. As to the conception and attitude toward the profession of nursing according to benefits by scholarships, those who were benefitted by scholarship showed stronger conception of profession than those who did not receive scholarship and there was a remarkable difference between these two categories. However, there was no remarkable difference between these two categories in the extent of interest of the activities of nursing fields and in the reading of Code for Nurses. 13. As activation for study according to the benefits of scholarships, those who were benefitted by scholarships showed stronger response to the motive for study comparing with those who receive. 14. As to tile field of reading according to the benefits by scholarships, those who received scholarships tended to read autobiographies and essayers to a considerable extent. Those who did not receive scholarships tended to read novels. Those who received scholarships more read nursing boots than those who did not receive scholarships. 15. As to the attitude of study and doing of assignment according to benefits of scholarships, those who received scholarships managed a favorable life with schedules for study, More students of receiving scholarship showed a regular study for more than one hour per day. Also, in the method of doing home tasks, more students of receiving scholarship showed reference to relative books frequently for the submission of completed assignments.

• Molecules and Cells
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• 제26권5호
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• pp.443-453
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• 2008

The Bric-a-brac, Tramtrack, Broad-complex (BTB) domain is a protein-protein interaction domain that is found in many zinc finger transcription factors. BTB containing proteins play important roles in a variety of cellular functions including regulation of transcription, regulation of the cytoskeleton, protein ubiquitination, angiogenesis, and apoptosis. Here, we report the cloning and characterization of a novel human gene, KLHL31, from a human embryonic heart cDNA library. The cDNA of KLHL31 is 5743 bp long, encoding a protein product of 634 amino acids containing a BTB domain. The protein is highly conserved across different species. Western blot analysis indicates that the KLHL31 protein is abundantly expressed in both embryonic skeletal and heart tissue. In COS-7 cells, KLHL31 proteins are localized to both the nucleus and the cytoplasm. In primary cultures of nascent mouse cardiomyocytes, the majority of endogenous KLHL31 proteins are localized to the cytoplasm. KLHL31 acts as a transcription repressor when fused to GAL4 DNA-binding domain and deletion analysis indicates that the BTB domain is the main region responsible for this repression. Overexpression of KLHL31 in COS-7 cells inhibits the transcriptional activities of both the TPA-response element (TRE) and serum response element (SRE). KLHL31 also significantly reduces JNK activation leading to decreased phosphorylation and protein levels of the JNK target c-Jun in both COS-7 and Hela cells. These results suggest that KLHL31 protein may act as a new transcriptional repressor in MAPK/JNK signaling pathway to regulate cellular functions.