• Title/Summary/Keyword: life science domain

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Differential Intracellular Localization of Mitotic Centromere-associated Kinesin (MCAK) During Cell Cycle Progression in Human Jurkat T Cells (인체 Jurkat T 세포에 있어서 세포주기에 따른 MCAK 단백질의 세포 내 위치변화)

  • Jun Do Youn;Rue Seok Woo;Kim Su-Jung;Kim Young Ho
    • Journal of Life Science
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    • v.15 no.2 s.69
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    • pp.253-260
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    • 2005
  • Mitotic centromere-associated kinesin (MCAK), which is a member of the Kin I (internal motor domain) subfamily of kinesin-related proteins, is known to play a role in mitotic segregation of chromosome during M phase of the cell cycle. In the present study, we have produced a rat polyclonal antibody using human MCAK (HsMCAK) expressed in E. coli as the antigen. The antibody specifically recognized the HsMCAK protein (81 kDa), and could detect its nuclear localization in human Jurkat T cells and 293T cells by Western blot analysis. The specific stage of the cell cycle was obtained through blocking by either hydroxyl urea or nocodazole and subsequent releasing from each blocking for 2, 4, and 7 h. While the protein level of HsMCAK reached a maximum level in the S phase with slight decline in the $G_{2}-M$ phase, the electrophoretic mobility shift from $p81^{MCAK}\;to\;p84^{MCAK}$ began to be induced in the late S phase and reached a maximum level in the $G_{2}/M $ phase, and then it disappeared as the cells enter into the $G_{1}$ phase. Immunocytochemical analysis revealed that HsMCAK protein localized to centrosome and nucleus at the interphase, whereas it appeared to localize to the spindle pole, centromere of the condensed mitotic DNA, spindle fiber, or midbody, depending on the specific stage of the M phase. These results demonstrate that a rat polyclonal antibody raised against recombinant HsMCAK expressed in E. coli specifically detects human MCAK, and indicate that the electrophoretic mobility shift from $p81^{MCAK}\;to\;p84^{MCAK}$, which may be associated with its differential intracellular localization during the cell cycle, fluctuates with a maximum level of the shift at the $G_{2}-M$ phase.

Studies on the Function of the Rv2435c Gene of the Mycobacterium bovis BCG (Mycobacterium bovis BCG Rv2435c 유전자의 기능에 대한 연구)

  • Lee Seung-Sil;Bae Young-Min
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.415-422
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    • 2005
  • library of the mutants was prepared by transposon mutagenesis of the Mycobacterium bovis BCG. We screened this library for the resistance to an anti-tuberculosis antibiotic, PA-824. Most of the mutants resistant to the PA-824 were not able to synthesize the coenzyme $F_{420}$ which is normally produced by the wild type M. bovis BCG strains. HPLC analysis of the cellular extract showed that one of those mutants which lost the ability to synthesize $F_{420}$ still produced F0. The insertion site of the transposon in this mutant was determined by an inverse PCR and the transposon was found to be inserted in the Rv2435c open reading frame (ORF). Rv2435c ORF is predicted to encode an 80.3 kDa protein. Rv2435c protein appears to be bound to the cytoplasmic membrane, its N-terminal present in the periplasm and C-terminal in the cytoplasm. The C-terminal portion of this protein is highly homologous with the adenylyl cyclases of both prokaryotes and eukaryotes. There are 15 ORFs which have homology with the class III AC proteins in the genome of the M. tuberculosis and M. bovis. Two of those, Rv1625c and Rv2435c, are highly homologous with the mammalian ACs. We cloned the cytoplasmic domain of the Rv2435c ORF and expressed it with six histidine residues attached on its C-terminal in Escherichia coli to find out if this protein is a genuine AC. Production of that protein in E. coli was proved by purifying the histidine-tagged protein by using the Ni-NTA resin. This protein, however, failed to complement the cya mutation in E. coli, indicating that this protein lacks the AC activity. All of the further attempts to convert this protein to a functional AC by a mutagenesis with UV or hydroxylamine, or construction of several different fusion proteins with Rv1625c failed. It is, therefore, possible that Rv2435c protein might affect the conversion of F0 to $F_{420}$ not by synthesizing cAMP but by some other way.

Study on the Storage of Chestnut (밤 저장(貯藏)에 관(關)한 연구(硏究))

  • Yim, Ho;Kim, Choung-Ok;Shin, Dang-Wha;Suh, Kee- Bong
    • Korean Journal of Food Science and Technology
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    • v.12 no.3
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    • pp.170-175
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    • 1980
  • A mass production of chestnut necessiates the development of economic long-term storage method. The main objective of this study was to confirm the technical aspect of the chestnut storage method which was developed by two year project and to review the method of commercial application. The chestnut used for the experiments were separated in brine $(5.5{\sim}6.0^{\circ}\:B{\acute{a}}ume)$ into matured and unmatured lots and fumigated with $CS_2$ at a 5 $lb/27\;m^3$ level for $25{\sim}30\;hrs.$ The chestnuts were packed in wooden boxes with sawdust (50% moisture) in the ratio of 1 : 1 by volume. The boxes were stored in the cold room $(1{\pm}1^{\circ}C,\;85{\sim}95%\;RH)$ and the cellar ($0{\sim}10^{\circ}C$, controlled only by circulating night cool air). The results obtained were as follows: 1. Fully matured chestnut could be successfully preserved $8{\sim}9\;months$ at a l0% decay level in the cold room and $4{\sim}5\;months$ months in cellar. 2. Immatured chestnuts wire inferior to the matured in storage stability. At the maximum storage period, its storage life was two months shorter. 3. The heat transfer equation of piled chestnuts with sawdust can be suggested as $T_{\infty}-T_0=(T_{\infty}-T_0){\cdot}10^{-t/320}$ and j and $f_h$ values were 1 and 320 min, respectively. 4. The chestnuts in the package of storage unit had longer shelf life than naked chestnut during the retail distribution at ambient temperature.

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Ferritin, an Iron Storage Protein, Associates with Kinesin 1 through the Cargo-binding Region of Kinesin Heavy Chains (KHCs) (철 저장 단백질 ferritin과 kinesin 1 결합 규명)

  • Jang, Won Hee;Jeong, Young Joo;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.26 no.6
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    • pp.698-704
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    • 2016
  • The intracellular transport of organelles and protein complexes is mediated by kinesin superfamily proteins (KIFs). The first kinesin, kinesin 1, was identified as a molecular motor protein that moves various organelles and protein complexes along the microtubule rails in cells. Kinesin 1 is a tetramer of two heavy chains (KHCs, also called KIF5s) and two kinesin light chains (KLCs). KIF5s interact with many different proteins through their tail region, but their binding proteins have not yet been fully identified. To identify the interaction proteins for KIF5A, we performed yeast two-hybrid screening and found a specific interaction with ferritin heavy chain (Frt-h), which has a role in iron storage and detoxification. Frt-h bound to the amino acid residues between 800 and 940 of KIF5A and to other KIF5s in the yeast two-hybrid assay. The coiled-coil domain of Frt-h is essential for interaction with KIF5A. In addition, ferritin light chain (Frt-l) interacted with KIF5s in the yeast two-hybrid assay. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to KHC specifically co-immunoprecipitated Frt-h and Frt-l from mouse brain extracts. These results suggest the kinesin 1 motor protein may transport the ferritin complex in cells.

Transcriptome Analyses for the Anti-Adipogenic Mechanism of an Herbal Composition (생약복합물의 지방세포형성억제 기전규명을 위한 전사체 분석)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Bae, Sung-Min;Chae, Soo-Ahn;Lee, Jung-Ju;Oh, Dong-Jin;Park, Suk-Won;Cho, Soo-Hyun;Shim, Yae-Jie;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1054-1065
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    • 2010
  • SH21B is a natural composition composed of seven herbs: Scutellaria baicalensis Georgi, Prunus armeniaca Maxim, Ephedra sinica Stapf, Acorus gramineus Soland, Typha orientalis Presl, Polygala tenuifolia Willd and Nelumbo nucifera Gaertner (Ratio 3:3:3:3:3:2:2). In our previous study, we reported that SH21B inhibited adipogenesis and fat accumulation in 3T3-L1 cells through modulation of various regulators in the adipogenesis pathway. The aim of this study was to analyze the transcriptome profiles for the anti-adipogenic effects of SH21B in 3T3-L1 cells. Total RNAs from SH21B-treated 3T3-L1 cells were reverse-transcribed into cDNAs and hybridized to Affymetrix Mouse Gene 1.0 ST array. From microarray analyses, we identified 2,568 genes of which expressions were changed more than two-fold by SH21B, and the clustering analyses of these genes resulted in 9 clusters. Three clusters among the 9 showed down-regulation by SH21B (cluster 4, cluster 6 and cluster 9), and two clusters showed up-regulation by SH21B (cluster 7 and cluster 8) during the adipogenesis of 3T3-L1 cells. It was found that many genes related to cell proliferation and adipogenesis were included in these clusters. Clusters 4, 6 and 9 included genes which were related with adipogenesis induction and cell cycle arrest. Clusters 7 and 8 included genes related to cell proliferation as well as adipogenesis inhibition. These results suggest that the mechanisms of the anti-adipogenic effects of SH21B may be the modulation of genes involved in cell proliferation and adipogenesis.

Limitations on Exclusive Rights of Authors for Library Reprography : A Comparative Examination of the Draft Revision of Korean Copyright Law with the New American Copyright Act of 1976 (저작권법에 준한 도서관봉사에 관한 연구 -미국과 한국의 저자재산권의 제한규정을 중시으로-)

  • 김향신
    • Journal of Korean Library and Information Science Society
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    • v.11
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    • pp.69-99
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    • 1984
  • A dramatic development in the new technology of copying materials has presented us with massive problems on reconciling the conflicts between copyright owners and potential users of copyrighted materials. The adaptation to this changing condition led some countries to revise their copyright laws such as in the U. S. in 1976 and in Korea in 1984 for merging with the international or universal copyright conventions in the future. Copyright defined as exclusive rights given to copyright owners aims to secure a fair return for an author's creative labor and to stimulate artistic creativity for the general public good. The exclusive rights on copyrightable matters, generally for reproduction, preparation of derivative works, public distribution, public performance, and public display, are limited by fair use for scholarship and criticism and by library reproduction for its preservation and interlibrary loan. These limitations on the exclusive rights are concerned with all aspects of library services and cause a great burden on librarian's daily duty to provide balance between the rights of creators and the needs of library patrons. The fair use as one of the limitations on it has been coupled with enormous growth of a new technology and extended from xerography to online database systems. The implementation of the fair use and library reprography in Korean law to the local practices is examined on the basis of the new American copyright act of 1976. Under the draft revision of Korean law, librarians will face many potential problems as summarized below. 1. Because the new provision of 'life time plus 50 years' will tie up substantial bodies of material longer than the old law, until that date librarians would need permissions from the owners and should pay attention to the author's death date. 2. Because the copyright can be sold, distributed, given to the heirs, donated, as a whole or a part, librarians should chase down the heirs and other second owners. In case of a derivative work, this is a real problem. 3. Since a work has its protection from the moment of its creation, the coverage of copyrightable matter would be extended to the published or the unpublished works and librarian's work load would be heavier. Without copyright registration, no one can be certain that a work is in the public domain. Therefore, librarians will need to check with an authority. 4. For implementation of limitations on exclusive rights, fair use and library reproduction for interlibrary loan, there can be no substantial aggregate use and there can be no systematic distribution of multicopies. Therefore, librarians should not substitute reproductions for subscriptions or purchases. 5. For the interlibrary loan by photocopying, librarians should understand the procedure of royalty payment. 6. Compulsory licenses should be understood by librarians. 7. Because the draft revision of Korean law is a reciprocal treaty, librarians should take care of other countries' copyright law to protect foreign authors from Korean law. In order to solve the above problems, some suggestions are presented below. 1. That copyright clearinghouse or central agency as a centralized royalty payment mechanism be established. 2. That the Korean Library Association establish a committee on copyright. 3. That the Korean Library Association propose guidelines for each occasion, e.g. for interlibrary loan, books and periodicals and music, etc. 4. That the Korean government establish a copyright office or an official organization for copyright control other than the copyright committee already organized by the government. 5. That the Korean Library Association establish educational programs on copyright for librarians through seminars or articles written in its magazines. 6. That individual libraries provide librarian's copyright kits. 7. That school libraries distribute subject bibliographies on copyright law to teachers. However, librarians should keep in mind that limitations on exclusive rights are not for an exemption from library reprography but as a convenient access to library resources.

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An Ethnographic Study about Taegyo Practice in Korea (태교 실천에 대한 일상생활 기술적 연구)

  • 김현옥
    • Journal of Korean Academy of Nursing
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    • v.27 no.2
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    • pp.411-422
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    • 1997
  • The purpose of this study is twofold : (i) to investigate how much effort the married couples are making for the good health of both the pregnant woman and her unborn child from the time of their marriage to and during the period of conception : and (ii) to comprehensive investigate socio-cultural back-grounds which affect prenatal effort. Result of this study provide a basis for the prenatal care program which will be appropriate to our culture. This study has been done by the ethnographic research method. The subjects of this study are 53 people in all consisting of 33 pregnant women and 20 husbands. In order to investigate socio-cultural factors which influence Taegyo, producers of Taegyo music were interviewed. In addition the researcher surveyed the markets of Taegyo music, participated in special courses of prenatal education, analyzed the content of the books and periodicals dealing with Taegyo, and collected the concept of Taegyo distributed by the mass media. The full-fledged study continued for eight months from February to August.1996. The data were analyzed as soon as they were collected. Spradly's(1979, 1980) developmental, sequential method of domain analysis. taxonomic analysis, componential analysis, and theme analysis in this order was adopted as the procedure of analyzing the data. To obtain the exactness of study, Sandelowski's (1986) four criteria, that is, Credibility, Fittingness, Auditability, and Confirmability were applied to all stages of data collection, data analysis, the interpretation of the result, and the description of the result. The following are the result : 1. The couples' Taegyo at the stage of preconception was related to their physical, psychological, spiritual conditions under which a healthy baby will be born. Specific methods they prefer are : "the choice of one's spouse." "physical check-up," "physical good health, " "praying, " and so on. 2. When the marriod couple have sex in order to conceive, their Taegyo was related to the imposition of their physical, psychological, and environmental conditions. Specific methods they prefer are : "having sex at specific time, " "having sex in nice place." "to purify their minds while having sex," and so on. 3. The married couples' Taegyo while they are in pregnancy was related to the imposition of their physical. psychological, emotionmental. environmental, social and spiritual conditions. Specific methods they prefer are : "listening to music. " "reading," "looking at beautiful things only," "to avoid looking at or listening to bad things." "to eat food in good shape, " "to avoid drugs," "eating Korean herbal medicine." "sexual abstinence," "to avoid dangerous places," "to keep emotional tranquility," "moderate exercises and rest." "leading a pure life." "praying." "being aware of their words and behavior." "for the couple to keep a good relationship." "interaction with their unborn child," "to support Taegyo for pregnant women," and so on. 4. The married couple put Taegyo into practice on the basis of the following principles : the principle of respecting an unborn child, the principle of forming a good disposition. the principle of top-down parental love, the principle of synergy between a pregnant woman and her unborn child, the principle of expecting a good child, the principle of forming a good habit, and the principle of acquiring a parental role. 5. The practice of Taegyo is influenced by such factors as the married couple, the supporting system, and the mass media. As the husband -and-wife factor, their information of Taegyo, the degree of importance is assigned to their characters, their time to spare, their healthiness, the age of pregnant woman, their conception plan, their religion, their belief of the Taegyo effects, and the birth of a baby in this order. The factor of the supporting system consists of her husband's support, her family support, and her neighbor's support. The mass media factors include the broadcasting media, books specialized in Taegyo, periodicals for pregnant women, booklets for advertizing powdered milk, Taegyo music of record manufacturing companies, and the teaching materials for gifted children. Among these the mass media is especially taking advantage of Taegyo as its main source of economic profits are leading the public behavior pattern to a prodigal one. Taegyo is a self-control behavior which requires practice for the following : the physical and psychological good health of the pregnant woman and her unborn child, the development of the unborn child's good character, the development of the unborn child's intelligence and talents, the expectation of the unborn child's good features. shape a good habit, the expectation of the unborn child's bright future, and the learning of a parental role, the expectation of male birth. Above all it is a type of our good cultural tradition which pursues a value higher than the one that the prenatal care does. The principles of pregnancy care inherent in the habit of Taegyo will provide us a guideline for the development of the prenatal care.

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Physiological and Proteomic Responses of Barley Seedlings to Salt Stress (보리의 생육초기 염 스트레스에 따른 생리적 반응과 프로테옴 변화)

  • Kim, Dea-Wook;Yun, Seong-Kun;Park, Hyoung-Ho;Hwang, Jong-Jin;Han, Ok-Kyu;Park, Tae-Il;Jung, Gun-Ho;Lee, Jae-Eun;Kim, Sun-Lim;Chung, Young-Ho
    • Journal of the Korean Society of International Agriculture
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    • v.23 no.5
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    • pp.537-545
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    • 2011
  • This study was conducted to obtain basic information on physiological and proteomic responses of barley seedlings to salt stress. Shoot dry weight decreased significantly as the level of soil salinity increased. Salt stress-induced decrease of relative shoot dry weight was lower in cv. "Sanglok" than in cv. "Sunwoo". Under the salt stress, SPAD value decreased, and the value was higher in cv. "Sanglok" than in cv. "Sunwoo". Sodium ion content in the leaves increased as NaCl concentration increased, and the content was higher in cv. "Sunwoo" than in cv. "Sanglok". The K+/Na+ ratio was higher in cv. "Sanglok" than in cv. "Sunwoo". Salt stress-induced alterations in protein expression of the leaves were detected by two dimensional electrophoresis, and 47 protein spots showing altered expression were selected. Among the selected protein spots, 17 protein spots were up-regulated and 28 spots down-regulated in cv. "Sanglok". In cv. "Sunwoo", 14 protein spots were up-regulated and 27 spots down-regulated. Out of 47 deferentially expressed protein spots, 18 protein spots were identified using mass spectrometry and NCBI protein database. Among the identified proteins, ten proteins are known to be involved in various stress responses, but the others are not directly involved in stress responses.