• Title/Summary/Keyword: $2^{n}-translation$

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A Splice Variant of the C2H2-Type Zinc Finger Protein, ZNF268s, Regulates NF-κB Activation by TNF-α

  • Chun, Jung Nyeo;Song, In Sung;Kang, Dong-Hoon;Song, Hye Jin;Kim, Hye In;Suh, Ja Won;Lee, Kong Ju;Kim, Jaesang;Won, Sang
    • Molecules and Cells
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    • v.26 no.2
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    • pp.175-180
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    • 2008
  • $I{\kappa}B$ kinase (IKK), the pivotal kinase in signal-dependent activation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$), is composed of multiple protein components, including IKK ${\alpha}/{\beta}/{\gamma}$ core subunits. To investigate the regulation of the IKK complex, we immunoaffinity purified the IKK complex, and by MALDI-TOF mass spectrometry identified a splice variant of zinc finger protein 268 (ZNF268) as a novel IKKinteracting protein. Both the full-length and the spliced form of the ZNF268 protein were detected in a variety of mammalian tissues and cell lines. The genes were cloned and expressed by in vitro transcription/translation. Several deletion derivatives, such as KRAB domain (KRAB) on its own, the KRAB/spacer/4-zinc fingers (zF4), and the spacer/4-zinc fingers (zS4), were ectopically expressed in mammalian cells and exhibited had different subcellular locations. The KRAB-containing mutants were restricted to the nucleus, while zS4 was localized in the cytosol. TNF-${\alpha}$-induced NF-${\kappa}B$ activation was examined using these mutants and only zS4 was found to stimulate activation. Collectively, the results indicate that a spliced form of ZNF268 lacking the KRAB domain is located in the cytosol, where it seems to play a role in TNF-${\alpha}$-induced NF-${\kappa}B$ activation by interacting with the IKK complex.

Molecular Characterization of Ischemia-Responsive Protein 94 (irp94) Response to Unfolded Protein Responses in the Neuron

  • Kim Seung-Whan;Kwon Ki-Sang;Shin Kee-Sun;Kim Seung-Ho;Kwon O-Yu
    • Biomedical Science Letters
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    • v.12 no.2
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    • pp.81-89
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    • 2006
  • The ischemia-responsive 94 gene (irp94) encoding a 94 kDa endoplasmic reticulum resident protein was investigated its molecular properties associated with unfoled protein responses. First, the expression of irp94 mRNA was tested after the reperfusion of the transient forebrain ischemia induction at the central nervous system in three Mongolian gerbils. Second, irp94 expression in PC12 cells, which are derived from transplantable rat pheochromocytoma cultured in the DMEM media, was tested at transcriptional and translational levels. The half life of irp94 mRNA was also determined In PC12 cells. Last, the changes of irp94 mRNA expression were investigated by the addition of various ER stress inducible chemicals (A23187, BFA, tunicamycin, DTT and $H_2O_2$) and proteasome inhibitors, and heat shock. High level expression of irp94 mRNA was detected after 3 hours reperfusion in the both sites of the cerebral cortex and hippocampus of the gerbil brain. The main regulation of irp94 mRNA expression in PC 12 cells was determined at the transcriptional level. The half life of irp94 mRNA in PC12 cells was approximately 5 hours after the initial translation. The remarkable expression of irp94 mRNA was detected by the treatment of tunicamycin, which blocks glycosylation of newly synthesized polypeptides, and $H_2O_2$, which induces apoptosis. When PC12 cells were treated with the cytosol proteasome inhibitors such as ALLN (N-acetyl-leucyl-norleucinal) and MG 132 (methylguanidine), irp94 mRNA expression was increased. These results indicate that expression of irp94 was induced by ER stress including oxidation condition and glycosylation blocking in proteins. Expression of irp94 was increased when the cells were chased after heat shock, suggesting that irp94 may be involved in recovery rather than protection against ER stresses. In addition, irp94 expression was remarkably increased when cytosol proteasomes were inhibited by ALLN and MG 132, suggesting that irp94 plays an important role for maintaining the ERAD (endoplasmic reticulum associated degradation) function.

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Species, Planting Position and Scenic Utilization of 'Paulownia Tree(梧桐)' in the Traditional Garden (전통정원에서 '오동(梧桐)'의 수종, 식재 위치와 경관적 활용)

  • Hong, Hyoung-Soon
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.31 no.2
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    • pp.20-30
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    • 2013
  • 'Paulownia tree', one of a tree species which has been with human civilization, has been widely used as a garden plant. The purpose of this study is to investigate concrete species of 'paulownia tree,' which had been planted in Korean traditional garden, the position of plantation, and consider its usefulness therefrom. The results of the study can be summarized as follows. 1. As the result of considering old literatures as encyclopedia, agricultural book (農書), gardening book, etc., there's a difference in the description of 'paulownia tree' depending on the complier, as well, the classification of concrete species is quite ambiguous. Therefore, it judged a limit which is planted based on point of the compass is not apply to species of tress of paulownia tree. Merely, the point of suitability and evasion(宜忌) related to the plantation of 'paulownia tree' could be identified in "Jeungbosanrimgyeongje(增補山林經濟) and "Imwongyeongjeji(林園經濟志)", not "Sanrimgyeongje (山林經濟)". 2. It could be confirmed again through poetry and prose which describe old garden that the words such as 'O(梧)', 'Dong(桐)', 'Odong(梧桐)', etc. were used without significant division. However, it is supposed that the species 'Odong' which was actually adopted at the garden might be Catalpa as well including Korean Paulownia and Chinese parasol tree. 3. It is considered that the reference point of suitability and evasion(宜忌) regarding 'paulownia tree' plantation was not generally applied. That is, species of paulownia tree was not divided for planting according to direction, as well, they seemed to willingly plant paulownia trees nearby the house as well, e.g. front yard or nearby yard, etc. 4. The usefulness of paulownia tree as a garden plant of an old garden played a role of 'the messenger of fall,' emphasizing a sense of the season. 5. Paulownia tree has another usefulness as a tree which adds an Ephemeral landscape. Therefore, the ancient people considered 'paulownia tree' that goes with 'the moon' the best, and enjoyed the quaint beauty of those two are juxtaposed. Also, 'paulownia tree' was utilized as a tree which adds an atmosphere of a rainy day, such as enjoying the sound of rain dropping on the 'paulownia tree', etc. The limitation of this study is that the research was performed being restricted to the translation among lots of Chinese references. Later-on task of research is the necessity of a more in-depth study through the discover of new historical sources and the accumulation of translation outcome.

Effect of different levels of protein concentrates supplementation on the growth performance, plasma amino acids profile and mTOR cascade genes expression in early-weaned yak calves

  • Peng, Q.H.;Khan, N.A.;Xue, B.;Yan, T.H.;Wang, Z.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.2
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    • pp.218-224
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    • 2018
  • Objective: This study evaluated the effects of different levels of protein concentrate supplementation on the growth performance of yak calves, and correlated the growth rate to changes occurring in the plasma- amino acids, -insulin profile, and signaling activity of mammalian target of rapamycin (mTOR) cascade to characterize the mechanism through which the protein synthesis can be improved in early weaned yaks. Methods: For this study, 48 early (3 months old) weaned yak calves were selected, and assigned into four dietary treatments according to randomized complete block design. The four blocks were balanced for body weight and sex. The yaks were either grazed on natural pasture (control diet) in a single herd or the grazing yaks was supplemented with one of the three protein rich supplements containing low (17%; LP), medium (19%; MP), or high (21%; HP) levels of crude proteins for a period of 30 days. Results: Results showed that the average daily gain of calves increased (0.14 vs 0.23-0.26 kg; p<0.05) with protein concentrates supplementation. The concentration of plasma methionine increased (p<0.05; 8.6 vs $10.1-12.4{\mu}mol/L$), while those of serine and tyrosine did not change (p>0.05) when the grazing calves were supplemented with protein concentrates. Compared to control diet, the insulin level of calves increased (p<0.05; 1.86 vs $2.16-2.54{\mu}IU/mL$) with supplementation of protein concentrates. Addition of protein concentrates up-regulated (p<0.05) expression of mTOR-raptor, mammalian vacuolar protein sorting 34 homolog, the translational regulators eukaryotic translation initiation factor 4E binding protein 1, and S6 kinase 1 genes in both Longissimus dorsi and semitendinosus. In contrast, the expression of sequestosome 1 was down-regulated in the concentrate supplemented calves. Conclusion: Our results show that protein supplementation improves the growth performance of early weaned yak calves, and that plasma methionine and insulin concentrations were the key mediator for gene expression and protein deposition in the muscles.

Cloning of Gene Fragment having Homology with the Polypetide Chymotrypsin Inhibitor from the Potato Proteinase Inhibitor II Gene and Its Expression in E. coli. (감자 단백질 분해효소 억제제-II 유전자로부터의 폴리펩타이드 카이모트립신 저해제와 homology가 있는 유전자단편의 클로닝 및 대장균에서의 발현)

  • Jung, Jin;Park, Sang-Gyu
    • Applied Biological Chemistry
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    • v.38 no.5
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    • pp.382-386
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    • 1995
  • The potato proteinase inhibitor II (PI-II) protein contains chymotrypsin and trypsin inhibitory site. Among several PI-II genes isolated from genomic library, amino acid sequence deduced from PI-IIT gene has 84% identity with that of the polypeptide chymotrypsin inhibitor (PCI). Therefore a gene fragment having homology with the PCI was cloned into a vector using polymerase chain reaction(PCR) from the potato proteinase inhibitor IIT gene. Two different primers were utilized for cloning; primer A contains NdeI restriction site and 30 nucleotides, which has AUG N-terminal methionine codon, primer B contains BclI restriction site and 28 nucleotides, which has TAG translation stop codon. After PCR, about 160 bp-long DNA fragment was cloned into pRT146, derivative of pUC118, and sequenced. The sequenced NdeI/BclI fragment was moved to pET3a, containing bacteriophage T7 promoter and terminator. The expressed proteins in E. coli BL2l(DE3) were determined on a polyacrylamide gel containing sodium dodecyl sulfate. The expected size of protein deduced from the sequenced gene fragment is about 6,500 dalton whose size was similar to the IPTG-induced protein (6,000 dalton) on a gel. However the expression level was much lower than expected.

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D99 Type I Signal Peptidase Implicated Stabilizing the Protein Structure (Type I 신호펩디드 가수분해효소에 존재하는 D99 아미노산 잔기의 구조적 역할 가능성)

  • Sung, Meesook;Eunyoung Han;Lee, Hoyoung
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.140-144
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    • 2003
  • Type Ⅰ signal peptidase is an integral membrane protein that functions to cleave signal peptides from secreted and membrane proteins. The enzyme serves as a potential target for the development of novel antibacterial agents due to its unique physiological properties. Despite being one of the best characterized enzymes, the catalysis of Type Ⅰ signal peptidase still remains controversy over the catalytic serine/lysine dyad mechanism. It appears that the dyad proteases are generally less efficient than the prototypical serine/histidine/aspartic acid triad found in most enzymes, although Type Ⅰ signal peptidase is an exception to this rule. In this paper, we have proposed that Type Ⅰ signal peptidase may act as the serine/lysine/aspartic acid triad cataltytic mechanism. Therefore, the aspartic acid 99 residue in the E. coli signal peptidase was chosen and mutated to an alanine to see if there is any possible role of the aspartic acid in the catalytic function. Type Ⅰ signal peptidase D99A protein was inactive in vitro assay using the procoat synthesized by in vitro transcription translation. However, the mutant was active using a highly sensitive in vivo assay. Pulse-chase experiments show that the replacement of aspartic acid 99 with alanine results in a very unstable signal peptidase molecule. Therefore, we conclude that it is unlikely that the residue is directly involved in catalysis, but rather plays an important role in stabilizing the protein structure.

Function of Global Regulator CodY in Bacillus thuringiensis BMB171 by Comparative Proteomic Analysis

  • Qi, Mingxia;Mei, Fei;Wang, Hui;Sun, Ming;Wang, Gejiao;Yu, Ziniu;Je, Yeonho;Li, Mingshun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.152-161
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    • 2015
  • CodY is a highly conserved protein in low G+C gram-positive bacteria that regulates genes involved in sporulation and stationary-phase adaptation. Bacillus thuringiensis is a grampositive bacterium that forms spores and parasporal crystals during the stationary phase. To our knowledge, the regulatory mechanism of CodY in B. thuringiensis is unknown. To study the function of CodY protein in B. thuringiensis, BMB171codY- was constructed in a BMB171 strain. A shuttle vector containing the ORF of cry1Ac10 was transformed into BMB171 and BMB171codY-, named BMB171cry1Ac and BMB171codY-cry1Ac, respectively. Some morphological and physiological changes of codY mutant BMB171codY-cry1Ac were observed. A comparative proteomic analysis was conducted for both BMB171codY-cry1Ac and BMB171cry1Ac through two-dimensional gel electrophoresis and MALDI-TOF-MS/MS analysis. The results showed that the proteins regulated by CodY are involved in microbial metabolism, including branched-chain amino acid metabolism, carbohydrate metabolism, fatty acid metabolism, and energy metabolism. Furthermore, we found CodY to be involved in sporulation, biosynthesis of poly-β-hydroxybutyrate, growth, genetic competence, and translation. According to the analysis of differentially expressed proteins, and physiological characterization of the codY mutant, we performed bacterial one-hybrid and electrophoretic mobility shift assay experiments and confirmed the direct regulation of genes by CodY, specifically those involved in metabolism of branched-chain amino acids, ribosomal recycling factor FRR, and the late competence protein ComER. Our data establish the foundation for in-depth study of the regulation of CodY in B. thuringiensis, and also offer a potential biocatalyst for functions of CodY in other bacteria.

Compensation Analysis of Cell Delay Variation for ATM Transmission in the TDMA Method (TDMA 방식에서 ATM 전송을 위한 셀 지연 변이의 보상 해석)

  • Kim, Jeong-Ho;Choe, Gyeong-Su
    • The Transactions of the Korea Information Processing Society
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    • v.3 no.2
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    • pp.295-304
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    • 1996
  • Toprovide economical BISDN service, with which integration process of many types of media is possible, it is necessary to construct a system with ground network and satellite network combined. The method for this type of transmission using satellite is TDMA that can provide services to many users in various area. However, the most difficult task to connect TDMA which uses synchronous method to ATM which used asynchronous transfer mode is the deterioration n of ATM transmission quality such as cell delay variation. Therefore, it is necessary to develop delay variation compensation method which can confront to the ATM. Efficient ways to use satellite links under the conditions such that maximum efficiency of the delay variation is limited under the required value, and the burst characteristic of transmission cell does not increase are being researched for translation between in ATM and TDMA. This paper points out the problems when time stamp method, reviewd in ground network, is applied to the satellite links to compensate the delay variation .To solve the problem, discrete cell count method is introduced along with the calculation of transmission capacity and error rate.Also, from the observation of stab-ility of the system and verification of reliability even when singal error occurred in the cell transmission timing information, the proposed compensation method appeared to be excellent.

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Superoxide Dismutase Gene Expression in the Endotoxin-Treated Rat Lung (내독소에 의한 백서 폐장의 Superoxide Dismutase 유전자 발현에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.3
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    • pp.215-221
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    • 1994
  • Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

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A Study of the Attitudes of Nursing Students toward Their Clinical Affiliation in a Mental Hospital (정신과간호 실습에 대한 간호 학생들의 태도 조사연구)

  • 김소야자
    • Journal of Korean Academy of Nursing
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    • v.3 no.3
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    • pp.15-26
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    • 1973
  • (Directed by Professor Hong, Shin Yong) Today, over seventy five Percent of the schools of nursing in Korea Provide a psychiatric experience in the basic curriculum. The psychiatric presents numerous major problems of adjustment to the student. The importance of positive attitudes toward the nursing care of psychiatric patients is recognized by the nursing profession. The purpose of this study was to determine the expressed attitudes of fifty-three nursing students toward their psychiatric affiliation. An attempt, also, was made to determine what implications these attitudes revealed relative to future program planning for students during the psychiatric nursing affiliation. A questionnaire, a Korean translation of the "psychiatric Nursing Attitude Questionnaire" by Milder Elizabeth Fletcher, was administered to fifty-three nursing students from three schools of nursing in Seoul, Who had completed a four-week psychiatric affiliation in a large mental hospital during Mar. 19, 1973 to May 19, 1973. The questionnaire of 100 statements was administered in the following way: (1) Part 1, Preconceptions. was. given in individual conferences with each subject, during the first few days of their affiliation, and again during the final week of the affiliation. The responses to Part Ⅰ were oral. (2) Part Ⅱ , Expectations, Part Ⅱ, Personal Relations, Part Ⅳ, Personal Feelings, and Part V , Attitudes and Activities of Patients were given to all of the subjects in a group meeting during the second week of the affiliation, and again. during the fourth week at the termination of the affiliation. Responses to Parts B, B, n, and f, wire written. Each of the 100 statements of the questionnaire was considered to be either Positive or Negative. A favorable response was assigned the Positive value of land an unfavorable response was assigned the Negative value of O. The coefficient of correlation was computed between the two sets of scores for the fifty-three nursing students., The mean score, the standard deviation, and the differences in the means on each of the five parts of the questionnaire were computed and the relationships calculated by a t-test. The results. of the study were as follows: 1. There was no significant correlation between the two sets of scores for the fifty-three nursing students during the four-week psychiatric affiliation. (r=573) 2. There was no significant difference in the mean scores between the first and final tests for any of the five parts of the questionnaire. 3. The Part.1, Preconceptions, data indicated nursing students enter the psychiatric affiliation with certain attitudes and preconceptions toward tile psychiatric affiliation which affect their psychiatric nursing experience, 4. The Part Ⅰ, Expectations, data indicated inappropriate expectations of students related to lack of experience, Lack of pre-psychiatric affiliation orientation, lack of social understanding, and feelings of insecurity. 5. The Part Ⅲ, Personal relations, data indicated some students have negative attitudes in personal relations with normal people in respect to psychological security and social responsibilities. 6. The Part Ⅳ, Personal feelings, data indicated nursing students have psychological insecurity & inappropriateness. 7. The Part Ⅴ, Attitudes and activities of patients, data indicated nursing students have negative attitudes of fear and frustration due to the psychotic behavior of certain patients in certain situations. 8. The data indicated preconceptions are predominate in unfavorable attitudes of students toward psychiatric nursing affiliation. Further researches indicated in the following areas: 1. Because of the limited number of students in this study, similar studies should be performed with larger groups for further validation of the results. 2. Because of the findings concerning the influence of the opinions of people in close contact with the students, similar studies of the attitudes of the staff in nursing schools, attitudes of graduate nurses and attitudes of the public should be done to determine weakness and strengths of present programs.

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