• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1563-1570
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• 2010

To construct a highly sensitive detection system for endocrine disruptors (EDs), we have compared the activity of promoters with the n-alkane-inducible cytochrome P450 gene (ALK1), isocitrate lyase gene (ICL1), ribosomal protein S7 gene (RPS7), and the translation elongation factor-1${\alpha}$ gene (TEF1) for the heterologous gene in Yarrowia lipolytica. The promoters were introduced into the upstream of the lacZ or hERa reporter genes, respectively, and the activity was evaluated by ${\beta}$-galactosidase assay for lacZ and Western blot analysis for hER${\alpha}$. The expression analysis revealed that the ALK1 and ICL1 promoters were induced by n-decane and by EtOH, respectively. The constitutive promoter of RPS7 and TEF1 showed mostly a high level of expression in the presence of glucose and glycerol, respectively. In particular, the TEF1 promoter showed the highest ${\beta}$-galactosidase activity and a significant signal by Western blotting with the anti-estrogen receptor, compared with the other promoters. Moreover, the detection system was constructed with promoters linked to the upstream of the expression vector for the hER${\alpha}$ gene transformed into the Y. lipolytica with a chromosome-integrated lacZ reporter gene under the control of estrogen response elements (EREs). It was indicated that a combination of pTEF1p-hER${\alpha}$ and CXAU1-2XERE was the most effective system for the $E_2$-dependent induction of the ${\beta}$-galactosidase activity. This system showed the highest ${\beta}$-galactosidase activity at $10^{-6}\;M\;E_2$, and the activity could be detected at even the concentration of $10^{-10}\;M\;E_2$. As a result, we have constructed a strongly sensitive detection system with Y. lipolitica to evaluate recognized/suspected ED chemicals, such as natural/synthetic hormones, pesticides, and commercial chemicals. The results demonstrate the utility, sensitivity, and reproducibility of the system for identifying and characterizing environmental estrogens.

• Nutrition Research and Practice
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• v.18 no.1
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• pp.1-18
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• 2024

BACKGROUND/OBJECTIVES: Endoplasmic reticulum (ER) stress in adipose tissue causes an inflammatory response and leads to metabolic diseases. However, the association between vitamin D and adipose ER stress remains poorly understood. In this study, we investigated whether 1,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃) alleviates ER stress in adipocytes. MATERIALS/METHODS: 3T3-L1 cells were treated with different concentrations (i.e., 10-100 nM) of 1,25(OH)₂D₃ after or during differentiation (i.e., on day 0-7, 3-7, or 7). They were then incubated with thapsigargin (TG, 500 nM) for an additional 24 h to induce ER stress. Next, we measured the mRNA and protein levels of genes involved in unfold protein response (UPR) and adipogenesis using real-time polymerase chain reaction and western blotting and quantified the secreted protein levels of pro-inflammatory cytokines. Finally, the mRNA levels of UPR pathway genes were measured in adipocytes transfected with siRNA-targeting Vdr. RESULTS: Treatment with 1,25(OH)₂D₃ during various stages of adipocyte differentiation significantly inhibited ER stress induced by TG. In fully differentiated 3T3-L1 adipocytes, 1,25(OH)₂D₃ treatment suppressed mRNA levels of Ddit3, sXbp1, and Atf4 and decreased the secretion of monocyte chemoattractant protein-1, interleukin-6, and tumor necrosis factor-α. However, downregulation of the mRNA levels of Ddit3, sXbp1, and Atf4 following 1,25(OH)₂D₃ administration was not observed in Vdr-knockdown adipocytes. In addition, exposure of 3T3-L1 preadipocytes to 1,25(OH)₂D₃ inhibited transcription of Ddit3, sXbp1, Atf4, Bip, and Atf6 and reduced the p-alpha subunit of translation initiation factor 2 (eIF2α)/eIF2α and p-protein kinase RNA-like ER kinase (PERK)/PERK protein ratios. Furthermore, 1,25(OH)₂D₃ treatment before adipocyte differentiation reduced adipogenesis and the mRNA levels of adipogenic genes. CONCLUSIONS: Our data suggest that 1,25(OH)₂D₃ prevents TG-induced ER stress and inflammatory responses in mature adipocytes by downregulating UPR signaling via binding with Vdr. In addition, the inhibition of adipogenesis by vitamin D may contribute to the reduction of ER stress in adipocytes.

• Bulletin of the Korean Chemical Society
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• v.33 no.8
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• pp.2508-2512
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• 2012

Cold shock proteins (CSPs) are a family of proteins induced at low temperatures. CSPs bind to single-stranded nucleic acids through the ribonucleoprotein 1 and 2 (RNP 1 and 2) binding motifs. CSPs play an essential role in cold adaptation by regulating transcription and translation via molecular chaperones. The solution nuclear magnetic resonance (NMR) or X-ray crystal structures of several CSPs from various microorganisms have been determined, but structural characteristics of psychrophilic CSPs have not been studied. Therefore, we optimized the purification process to obtain highly pure Lm-Csp and determined the three-dimensional structure model of Lm-Csp by comparative homology modeling using MODELLER on the basis of the solution NMR structure of Bs-CspB. Lm-Csp consists of a ${\beta}$-barrel structure, which includes antiparallel ${\beta}$ strands (G4-N10, F15-I18, V26-H29, A46-D50, and P58-Q64). The template protein, Bs-CspB, shares a similar ${\beta}$ sheet structure and an identical chain fold to Lm-Csp. However, the sheets in Lm-Csp were much shorter than those of Bs-CspB. The Lm-Csp side chains, E2 and R20 form a salt bridge, thus, stabilizing the Lm-Csp structure. To evaluate the contribution of this ionic interaction as well as that of the hydrophobic patch on protein stability, we investigated the secondary structures of wild type and mutant protein (W8, F15, and R20) of Lm-Csp using circular dichroism (CD) spectroscopy. The results showed that solvent-exposed aromatic side chains as well as residues participating in ionic interactions are very important for structural stability. Further studies on the three-dimensional structure and dynamics of Lm-Csp using NMR spectroscopy are required.

• BMB Reports
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• v.35 no.3
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• pp.267-272
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• 2002

TNF-$\alpha$ elicits various responses including apoptosis, proliferation, and differentiation according to cell type. In neuronal PC12 cells, TNF-$\alpha$ induces moderate apoptosis while lipopolysarccaharide or trophic factor deprivation can potentiate apoptosis that is induced by TNF-$\alpha$. TNF-$\alpha$ initiates various signal transduction pathways leading to the activation of the caspase family, NF-${\kappa}B$, Jun N-terminal kinase, and p38 MAPK via the death domain that contains the TNF-$\alpha$ receptor. Inhibition of translation using cycloheximide greatly enhanced the apoptotic effect of TNF-$\alpha$. This implies that the induction of anti-apoptotic genes for survival by TNF-$\alpha$ may be able to protect PC12 cells from apoptosis. Accordingly, Bcl-2, an anti-apoptotic genes for survival by TNF-$\alpha$ may be able to protect PC12 cells from apoptosis. Accordingly, Bcl-2, an anti-apoptotic Bcl-2 family member, was highly expressed in response to TNF-$\alpha$. In this study, we examined the anti-apoptotic role of p38 MAPK that is activated by TNF-$\alpha$ in neuronal PC12 cells. The phosphorylation of p38 MAPK in response to TNF-$\alpha$ slowly increased and lasted several hours in the PC12 cell and DRG neuron. This specific inhibitor of p38 MAPK, SB202190, significantly enhanced the apoptosis that was induced by TNF-$\alpha$ in PC12 cells. This indicates that the activation of p38 MAPK could protect PC12 cells from apoptosis since there is no known role of p38 MAPK in resoonse to TNF-$\alpha$ in neuron. This discovery could be evidence for the neuroprotective role of the p38 MAPK.

• Proceedings of the Korean Society of Crop Science Conference
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• 2004.04a
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• pp.12-27
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• 2004

Thanks to spectacular advances in the techniques for identifying proteins separated by two-dimensional electrophoresis and in methods for large-scale analysis of proteome variations, proteomics is becoming an essential methodology in various fields of plant sciences. Plant proteomics would be most useful when combined with other functional genomics tools and approaches. A combination of microarray and proteomics analysis will indicate whether gene regulation is controlled at the level of transcription or translation and protein accumulation. In this review, we described the catalogues of the rice proteome which were constructed in our program, and functional characterization of some of these proteins was discussed. Mass-spectrometry is a most prevalent technique to identify rapidly a large of proteins in proteome analysis. However, the conventional Western blotting/sequencing technique us still used in many laboratories. As a first step to efficiently construct protein data-file in proteome analysis of major cereals, we have analyzed the N-terminal sequences of 100 rice embryo proteins and 70 wheat spike proteins separated by two-dimensional electrophoresis. Edman degradation revealed the N-terminal peptide sequences of only 31 rice proteins and 47 wheat proteins, suggesting that the rest of separated protein spots are N-terminally blocked. To efficiently determine the internal sequence of blocked proteins, we have developed a modified Cleveland peptide mapping method. Using this above method, the internal sequences of all blocked rice proteins (i. e., 69 proteins) were determined. Among these 100 rice proteins, thirty were proteins for which homologous sequence in the rice genome database could be identified. However, the rest of the proteins lacked homologous proteins. This appears to be consistent with the fact that about 30% of total rice cDNA have been deposited in the database. Also, the major proteins involved in the growth and development of rice can be identified using the proteome approach. Some of these proteins, including a calcium-binding protein that fumed out to be calreticulin, gibberellin-binding protein, which is ribulose-1,5-bisphosphate carboxylase/oxygenase activate in rice, and leginsulin-binding protein in soybean have functions in the signal transduction pathway. Proteomics is well suited not only to determine interaction between pairs of proteins, but also to identify multisubunit complexes. Currently, a protein-protein interaction database for plant proteins (http://genome .c .kanazawa-u.ac.jp/Y2H)could be a very useful tool for the plant research community. Recently, we are separated proteins from grain filling and seed maturation in rice to perform ESI-Q-TOF/MS and MALDI-TOF/MS. This experiment shows a possibility to easily and rapidly identify a number of 2-DE separated proteins of rice by ESI-Q-TOF/MS and MALDI-TOF/MS. Therefore, the Information thus obtained from the plant proteome would be helpful in predicting the function of the unknown proteins and would be useful in the plant molecular breeding. Also, information from our study could provide a venue to plant breeder and molecular biologist to design their research strategies precisely.

• KSBB Journal
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• v.22 no.5
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• pp.362-365
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• 2007

Cyanobacteria have been identified as one of the most promising group producing novel biochemically active natural products. Cyanobacteria are a very old group of prokaryotic organisms that produce very diverse secondary metabolites, especially non-ribosomal peptide and polyketide structures. Large multienzyme complexes which are responsible for the non-ribosomal biosynthesis of peptides are modular for the addition of a single amino acid. An activation of amino acid substrates results in an amino adenylate occuring via an adenylation domain (A-domain). A-domains are responsible for the recognition of amino acids as substrates within NP synthesis. The A-domain contains ten conserved motifs, A1 to A10. In this study, ten conserved motifs from A1 to A10 were checked regarding their amino acid sequence of the NRPS-module of Microcystis aeruginosa PCC 7806. The part of the amino acid sequence chosen was that which contained as many conserved motives as possible, and then these amino sequence were compared between other cyanobacteria to design a new degenerate primer. A new degenerate primer (A3/A7 primer) was designed to detect any putative NP synthetase region in unkwon cyanobacteria by a reverse translation of the conserved amino acid sequence and a search for cyanobacterial DNA bank.

• The Journal of Korean Medical History
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• v.27 no.1
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• pp.135-156
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• 2014

Through a simple translation and analysis of the "Su$\acute{s}$uta-saṃhit$\bar{a}$(The fascinating tales) S$\bar{u}$trasth$\bar{a}$na(The Introduction)", I have summarized the results of the research into the $\bar{A}$yurveda Herbal theory. 1. The title of chapter 40 is "dravya-rasa-guṇa-vip$\bar{a}$ka-vij$\tilde{n}$anīya(Medcation Taste Nature Effects Digestion)", and it states theoretical knowledge on materia medica. It says that 'matter' is the most important thing, and that it decides the curing effect. Although it does acknowledge the reason behind the doctors that emphasized things like 'taste', it mainly has a critical tone. The difference in philosophies and medical theory can be seen when we compare this to the "Caraka-saṃhit$\bar{a}$", 'theory on taste'. 2. The title of chapter 41 is "dravya-vi$\acute{s}$eṣa-vij$\tilde{n}$anīya(A Discussion on the Characteristics of Materia Medica)", and the contents of the chapter have a similar tone to the thoughts of the S$\bar{a}$mkhya(數論派). All 'matter' was created by the combination of the five elements of earth, water, fire, wind, and ether, and the predominant element makes it have an earth, water, fire, wind, or ether nature. Earth has an unmoving and down going nature, and makes the body strong. Water has an irrigating nature, and moistens the tissue and induces secretion and excretion. Fire has an upward-going nature, and improves sight and complexion. Wind relaxes and dries the body, and makes the mind and body agile. Ether gives flexibility, porosity, and candidness. The title of chapter 42 is "rasa-vi$\acute{s}$eṣa-vij$\tilde{n}\bar{a}$nīya(A Discussion on the Types of Rasa(taste))". "Rasa" is an important concept in $\bar{A}$yurveda. Sometimes it is translated as 'one of the seven types of bodily tissue' and seen as chyle, and sometimes it is seen as the tastes that can be felt with the tongue such as spicy, sweet, sour, bitter, salty, and astringent(澁). Volume 1, chapter 42 of the "Caraka-saṃhit$\bar{a}$" is dedicated to the "types of taste", and in chapter 26, it theorizes the concept of 'taste'.

• Journal of the Institute of Electronics Engineers of Korea SP
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• v.45 no.6
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• pp.27-37
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• 2008

Applications of 3D data increase with advancement of multimedia technique and contents, and it is necessary to manage and to retrieve for 3D data efficiently. In this paper, we propose a new method using the sliced shape which extracts efficiently a feature description for shape-based retrieval of 3D models. Since the feature descriptor of 3D model should be invariant to translation, rotation and scale for its model, normalization of models requires for 3D model retrieval system. This paper uses principal component analysis(PCA) method in order to normalize all the models. The proposed algorithm finds a direction of each axis by the PCA and creates orthogonal n planes in each axis. These planes are orthogonalized with each axis, and are used to extract sliced shape image. Sliced shape image is the 2D plane created by intersecting at between 3D model and these planes. The proposed feature descriptor is a distribution of Euclidean distances from center point of sliced shape image to its outline. A performed evaluation is used for average of the normalize modified retrieval rank(ANMRR) with a standard evaluation from MPEG-7. In our experimental results, we demonstrate that the proposed method is an efficient 3D model retrieval.

• The Journal of Korean Institute of Communications and Information Sciences
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• v.34 no.12C
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• pp.1167-1176
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• 2009

In This Paper, we introduce an efficient IFFT design technique named for transmitter of OFDM (Orthogonal Frequency Division Multiplexing) system. In OFDM system, a cyclic prefix is inserted in forepart of OFDM symbol to prevent ICI(Inter-channel Interference) and ISI (Inter-symbol Interference). Attaching cyclic prefix causes delay in storing and copying IFFT result. The proposed IFFT removes this delay because its output is cyclic shifted by the length of cyclic prefix. So we can make a complete OFDM symbol by just copying the forepart of IFFT output to the end. In many cases, the length of cyclic prefix is 1/2n of FFT size, and this IFFT does not require additional hardware complexity and it does not cause any performance degradation.

• Korean Journal of Cognitive Science
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• v.4 no.2
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• pp.29-51
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• 1994

This paper presents a new approach to describing the meanings of English relative clauses with the theoretical framework of Conceptual Structure Theory (henceforth CST)which builds on the pionerring work of Sowa.And this paper aims at proposing some extensions to his work. CST describes the conceptual structrures of sentences with conceptual graphs(henceforth CG). which have begun to be used as an intermediate language in natural language processing and machine translation of computer.CGs are composed of concept types and conceptual relation types. They are a system of logic for semantic representation of sentences. This paper focuses on showing the differences of the CGs according to the functions of English relative clauses. English relative clauses are divided into restrictive and nonrestrictive uses.And this paper describes a restrictive clause with a CG including a expression.which derives from the viewpoint of Montague-semantics and Nom-S Analysis.This paper deals mainly with the relative clauses of double restroction as an example of restrictive relative clauses.The description of a nonrestrictive relative clause does not need any-expression, for it doesn's involve the meaning of set.And this paper links the CG of an appositive relative clause,which is a kind of nonrestrictive clauses,to the concept of the antecedent in the main clause.The description of a nonrestrictive relative clause with adverbial meaning is strated with two CGs for the main clause and the relative clause.They are linked with an appropriate intersentential conceptual relation type according to the contextual realtions between them.This paper also presents a CG of a sentential relative clause,which gives a comment on the main clause.