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Transgenic Siberian Ginseng Cultured Cells That Produce High Levels of Human Lactoferrin (인체 락토페린 생산 형질전환 가시오갈피 배양세포)

  • Jo Seung-Hyun;Kwon Suk-Yoon;Kim Jae-Whune;Lee Ki-Teak;Kwak Sang-Soo;Lee Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.209-215
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    • 2005
  • Lactoferrin is an iron-binding glycoprotein with many biological roles, including the protection against microbial and virus infection, stimulation of the immune system. We developed the transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing the human lactoferrin (hLf) protein following Agrobacterium tumefaciens-mediated transformation. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to hLf cDNA under the control of an oxidative stress-inducible SWPA2 promoter was engineered. Transgenic Siberian ginseng cultured cells to produce a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analysis. ELISA and western blot analysis showed that full length-hLf protein was synthesized in the transgenic cells. The production of hLf increased proportionally to cell growth and reached a maximal (up to 3% of total soluble proteins) at the stationary phase. These results suggest that the transgenic Siberian ginseng cultured cells in this study will be biotechnologically useful for the commercial production of medicinal plant cell cultures to produce hLf protein.

Metabolic Engineering of the Brassinosteroid Biosynthetic Pathways (Brassinosteroid의 대사공학)

  • Lee, Mi-Ock;Song, Ki-Hong;Lee, Hyun-Kyung;Jung, Ji-Yoon;Choe, Vit-Nary;Choe, Sung-Hwa
    • Journal of Plant Biotechnology
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    • v.29 no.2
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    • pp.139-144
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    • 2002
  • Sterols play two major roles in plants: a bulk component in biological membranes and precursors of plant steroid hormones. Physiological effects of plant steroids, brassinosteroids (BRs), include cell elongation, cell division, stress tolerance, and senescence acceleration. Arabidopsis mutants that carry genetic defects in BR biosynthesis or its signaling display characteristic phenotypes, such as short robust inflorescences, dark-green round leaves, and sterility. Currently there are more than 100 dwarf mutants representing 7 genetic loci in Arabidopsis. Mutants of 6 loci, dwf1/dim1/cbb1, cpd/dwf3, dwf4, dwf5, det2/dwf6, dwf7 are rescued by exogenous application of BRs, whereas bri1/dwf2 shares phenotypes with the above 6 loci but are resistant to BRs. These suggest that the 6 loci are defective in BR biosynthesis, and the one locus is in BR signaling. Biochemical analyses, such as intermediate feeding tests, examining the levels of endogenous BR, and molecular cloning of the genes revealed that dwf7, dwf5, and dwf1 are defective in the three consecutive steps of sterol biosynthesis, from episterol to campesterol via 5-dehydroepisterol. Similarly, det2/dwf6, dwf4, and cpd /dwf3 were shown to be blocked in D$^4$reduction, 22a-hydroxylation, and 23 a-hydroxylation, respectively. A signaling mutant bril/dwf2 carries mutations in a Leucine-rich repeat receptor kinase. Interestingly, the bri1 mutant was shown to accumulate significant amount of BRs, suggesting that signaling and biosynthesis are dynamically coupled in Arabidopsis. Thus it is likely that transgenic plants over-expressing the rate-limiting step enzyme DWF4 as well as blocking its use by BRIl could dramatically increase the biosynthetic yield of BRs. When applied industrially, BRs will boost new sector of plant biotechnology because of its potential use as a precursor of human steroid hormones, a novel lead compound for cholesterol-lowering effects, and a various application in plant protection.

Recombinant Human Thioredoxin-1 Protects Macrophages from Oxidized Low-Density Lipoprotein-Induced Foam Cell Formation and Cell Apoptosis

  • Zhang, Hui;Liu, Qi;Lin, Jia-Le;Wang, Yu;Zhang, Ruo-Xi;Hou, Jing-Bo;Yu, Bo
    • Biomolecules & Therapeutics
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    • v.26 no.2
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    • pp.121-129
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    • 2018
  • Oxidized low-density lipoprotein (ox-LDL)-induced macrophage foam cell formation and apoptosis play critical roles in the pathogenesis of atherosclerosis. Thioredoxin-1 (Trx) is an antioxidant that potently protects various cells from oxidative stress-induced cell death. However, the protective effect of Trx on ox-LDL-induced macrophage foam cell formation and apoptosis has not been studied. This study aims to investigate the effect of recombinant human Trx (rhTrx) on ox-LDL-stimulated RAW264.7 macrophages and elucidate the possible mechanisms. RhTrx significantly inhibited ox-LDL-induced cholesterol accumulation and apoptosis in RAW264.7 macrophages. RhTrx also suppressed the ox-LDL-induced overproduction of lectin-like oxidized LDL receptor (LOX-1), Bax and activated caspase-3, but it increased the expression of Bcl-2. In addition, rhTrx markedly inhibited the ox-LDL-induced production of intracellular reactive oxygen species (ROS) and phosphorylation of p38 mitogen-activated protein kinases (MAPK). Furthermore, anisomycin (a p38 MAPK activator) abolished the protective effect of rhTrx on ox-LDL-stimulated RAW264.7 cells, and SB203580 (a p38 MAPK inhibitor) exerted a similar effect as rhTrx. Collectively, these findings indicate that rhTrx suppresses ox-LDL-stimulated foam cell formation and macrophage apoptosis by inhibiting ROS generation, p38 MAPK activation and LOX-1 expression. Therefore, we propose that rhTrx has therapeutic potential in the prevention and treatment of atherosclerosis.

The Process of Occupational Socialization of the Court Security Team (법원경비관리대의 직업사회화 과정 분석)

  • Park, Ok-Cheol;Kwon, Tae-Il
    • Journal of the Society of Disaster Information
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    • v.8 no.3
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    • pp.276-286
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    • 2012
  • The purposes of this study are to investigate various experience factors of the court security team's occupational socialization process and suggest basic materials which are necessary to establish educational service strategies of preliminary court security and guard. Therefore, in-depth interview and ethnographic study were conducted for 4 court securities whose career is more than 3 years. As the result, the process of occupational socialization process of the court security was divided into preparation period, adaptation period, conflict period and maturity period. In the preparation period, vision of university departments, acquisition of certificates and information of the court security team are the factors of basic stage for becoming court securities. In the adaptation period, they adapt themselves for playing their own roles sincerely and become professional manpower after being court securities, through the university educational program useful for practical duties, certificates for practical duties and occupational satisfaction. In the conflict period, they aware conflicts of the company and job stress, that court securities should experience, and endure them. In the maturity period, they are grown by one more step as court securities, through the motivation, job professionalism, occupational prospect and efforts of self-development.

A LONGITUDINAL STUDY ON THE WIDTH CHANGE OF ANTERIOR KERATINIZED GINGIVA DURING TRANSITIONAL PERIOD (치아 교환시기중 전치부 각화치은의 폭경변화에 관한 연구)

  • Kim, Kee-Hong;Kim, Byung-Ok;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.24 no.1
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    • pp.15-25
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    • 1994
  • Keratinized ginigva has clinical singificance in periodontal health because it plays important roles in resistance to mechanical trauma, to penetration of bacteria, and to tensional stress by muscle attachment. In order to investigate the width change of anterior keratinized gingvial during transitional period, the width of kerainized gingiva on anterior teeth was measured annually in elementary school children with deciduous dentition until the successive anterior teeth were completely erupted. The distance from the gingival margin on mid-portion of crown to mucogingival junction was measured by Boley gauge(Hu-Friedy, U.S.A.) and was recorded as the width of keratinized gingiva. The difference of the width of keratinized gingiva according to sex and dentition was analyzed statistically by Student t-test. Following results were obtained : 1. In deciduous dentition, it was shown that the width of keratinized gingiva on maxillary central incisors($3.28{\pm}0.83mm$) was the widest(p<0.01), and that on maxillary lateral incisors, mandibular central and lateral incisors was $2.69{\pm}0.86mm$, $2.51{\pm}0.71mm$, $2.43{\pm}0.68mm$, respectively. 2. In mixed dentition, it was shown that the width of keratinized gingiva on maxillary central incisors ($5.10{\pm}0.86$) was the widest(p<0.01), and that on maxillary lateral incisors, mandibular central and lateral incisors was $4.35{\pm}0.83mm$, $3.51{\pm}0.60mm$, $3.57{\pm}0.66mm$, respectively. 3. The width of anterior keratinized gingiva was significantly increased after the successive anterior teeth were completely erupted(p<0.001). The width of keratinized gingiva on maxillary central and lateral incisors, and mandibular central and lateral incisors was increased by $1.82{\pm}0.83mm$, $1.65{\pm}0.69mm$, $0.99{\pm}0.39mm$, and $1.14{\pm}0.98mm$, respectively. 4. There was no statistical significance in the difference of the width of anterior keratinized gingiva between male and female(p>0.05).

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Expression Study on the Scaffold Gene of CRL4 Complex in Rice (Oryza sativa L.) (벼에 존재하는 CRL4 복합체 scaffold 유전자의 발현 양상에 대한 연구)

  • Bae, Yoowon;Kim, Hani;Kim, Sang-Hoon;Lee, Jae-Hoon
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1132-1139
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    • 2018
  • The stability of diverse cellular proteins in eukaryotes is regulated via ubiquitination. Moreover, E3 ligase plays a crucial role in determining substrate specificity and transfers ubiquitins into the substrates during the ubiquitination process. As a type of multi-subunit E3 ligase, cullin4 (CUL4)-based E3 ligase (CRL4) complex is involved in a variety of cellular processes, such as hormonal and stress responses in plants. In spite of several reports on the versatile roles of CRL4 in various signalings in Arabidopsis, CRL4's function in rice has been poorly known. To learn about CRL4-mediated cellular processes in rice in more detail, OsCUL4 that exhibits the highest homology with Arabidopsis CUL4 was isolated, and its expression patterns in various tissues and in response to plant hormones and abiotic stresses were monitored. Exogenous application of ABA or cytokinin increased the transcript levels of the OsCUL4 gene. Moreover, OsCUL4 was significantly upregulated in response to drought and salt stresses. These findings imply that OsCUL4 may be functionally related to ABA- and/or cytokinin-mediated cellular responses. OsCUL4 directly interacted with OsDDB1, an adaptor protein of CRL4, indicating that OsCUL4 can act as a scaffold protein of CRL4. An expression study on the OsCUL4 gene from this report could be used as a starting point to elucidate cellular responses in which a CRL4-mediated ubiquitination process is involved in rice.

A Study on the Planning Characteristics of Training Facilities Complex - Focusing on Training Facilities Planned through the Domestic Competitions after 2000s - (연수시설 단지의 계획특성 연구 - 2000년대 이후 국내 현상공모를 통해 계획된 연수시설을 중심으로 -)

  • Park, Hoon
    • Journal of the Korean Institute of Educational Facilities
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    • v.25 no.1
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    • pp.13-24
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    • 2018
  • Planning and designing training facilities has been developed from educational facilities, and increasingly diversified society has raised a need for facilities dedicated to training and education in a differentiated space separate from the one for regular works. This particular need of our times has led to the expansion of training facilities nationwide although they have something to be desired when it comes to planning and designing with sustainability associated with the locational characteristics of urban space as well as the unique types of facilities taken into account. Against this backdrop, this study will examined a variety of training facilities that have been established since 2000 through theoretical review and conduct intensive analysis on the characteristics of the planning aspects to suggest their significance and implications and to present the overall meanings and ramifications of planning approaches in consideration of new challenges modern training facilities are faced with, which have been revealed through architectural design competitions in recent years. The relevant implications are as follows. First, one of the locational advantages of training facilities, which is commanding beautiful scenery of the surrounding area, can be considered as intent to stress the aspect of a resort, one of the functions of any training facilities. As this study has demonstrated, many training facilities are located near around a beach or a lake. Second, training facilities can be classified into three different types in terms of their location: urban, suburban and resort and such locational characteristics are directly related to intended programs and differentiated links with target users. Third, the architectural styles of training institutes are differentiated in terms of harmonious arrangement between beautiful natural scenery and buildings in consideration of the layout characteristics of major facilities and the distance of ramps in and out of the facilities along with architectural features, including the transparency of building elevation and the type of slopes of roof structure. Fourth, the individual lodging buildings feature a variety of types depending on the design concept with different roles depending on the directional aspects such as the connection of ramps and the relations with the outside. Fifth, outdoor space plans are differentiated according to the intended purpose of training facilities. When it comes to gym facilities, for example, different outdoor space plans are found to be made depending on the original design concept such as outdoor playground-centered planning or golf facilities.

Effects of Mutation at Two Conserved Aspartate Residues and a Serine Residue on Functions of Yeast TSA 1 (Saccharomyces cerevisiae TSA1의 보존된 아스파트산 잔기 및 세린 잔기의 변이가 과산화효소 활성 및 샤페론 활성에 미치는 영향)

  • Lee, Songmi;Cho, Eun Yi;Kim, Kanghwa
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.81-86
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    • 2017
  • Alignment of 967 reference sequences of the typical 2-Cys peroxiredoxin family of proteins revealed that 10 amino acids were conserved, with over 99% identity. To investigate whether the conserved aspartic acid residues and serine residue affect the peroxidase and chaperone activity of the protein, we prepared yeast TSA1 mutant proteins in which aspartic acids at positions 75 and 103 were replaced by valine or asparagine, and serine at position 73 was replaced by alanine. By non-reducing SDS-PAGE, TSA1 and the S73A, D75V and D75N mutants were detected in dimeric form, whereas the D103V and D103N mutants were detected in various forms, ranging from high molecular-weight to monomeric. Compared with wild type TSA1, the D75N mutant exhibited 50% thioredoxin peroxidase activity, and the S73A and D75V mutants showed 25% activity. However, the D103V and D103N mutants showed no peroxidase activity. All proteins, except for the D103V and D103N mutants, exhibited chaperone activity at $43^{\circ}C$. Our results suggest that the two conserved aspartic acid residues and serine residue of TSA1 play important roles in its thioredoxin peroxidase activity, and D103 plays a critical role in its chaperone activity.

Arabidopsis MAP3K16 and Other Salt-Inducible MAP3Ks Regulate ABA Response Redundantly

  • Choi, Seo-wha;Lee, Seul-bee;Na, Yeon-ju;Jeung, Sun-geum;Kim, Soo Young
    • Molecules and Cells
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    • v.40 no.3
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    • pp.230-242
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    • 2017
  • In the Arabidopsis genome, approximately 80 MAP3Ks (mitogen-activated protein kinase kinase kinases) have been identified. However, only a few of them have been characterized, and the functions of most MAP3Ks are largely unknown. In this paper, we report the function of MAP3K16 and several other MAP3Ks, MAP3K14/15/17/18, whose expression is salt-inducible. We prepared MAP3K16 overexpression (OX) lines and analyzed their phenotypes. The result showed that the transgenic plants were ABA-insensitive during seed germination and cotyledon greening stage but their root growth was ABA-hypersensitive. The OX lines were more susceptible to water-deficit condition at later growth stage in soil. A MAP3K16 knockout (KO) line, on the other hand, exhibited opposite phenotypes. In similar transgenic analyses, we found that MAP3K14/15/17/18 OX and KO lines displayed similar phenotypes to those of MA3K16, suggesting the functional redundancy among them. MAP3K16 possesses in vitro kinase activity, and we carried out two-hybrid analyses to identify MAP3K16 substrates. Our results indicate that MAP3K16 interacts with MKK3 and the negative regulator of ABA response, ABR1, in yeast. Furthermore, MAP3K16 recombinant protein could phosphorylate MKK3 and ABR1, suggesting that they might be MAP3K16 substrates. Collectively, our results demonstrate that MAP3K16 and MAP3K14/15/17/18 are involved in ABA response, playing negative or positive roles depending on developmental stage and that MAP3K16 may function via MKK3 and ABR1.

The Analysis of Regional Stressor Level on Personal Factor and Reaction According of Radiological Technologist (방사선사의 개인요인과 반응에 따른 지역별 스트레스원 수준 분석)

  • Jung Hong-Ryang;Son Bu-Soon;Lim Cheong-Hwan
    • Journal of Environmental Health Sciences
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    • v.31 no.4 s.85
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    • pp.266-279
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    • 2005
  • The present study was conducted to analyze the causes of stress on the part of Korean radiological technologists depending on their working environments. For this purpose, 890 questionnaires were distributed to and collected from radiological technologists who were working at 44 general hospitals in 16 cities and provinces of Korea during the period from the mid July to the end of August 2003. The collected data were compared between regions, divided into the capital, metropolitan, medium and small cities, whose results are as follows: 1. As factors affecting working conditions, drinking was found to exert more influence on radiological technologists in small- and medium-sized cities, while loyalty to immediate seniors and organizations was the one producing greater influence on them in metropolitan cities than other cities. Overall, position, job satisfaction and physical condition seemed to have a great effect on them (p<0.001).2. In terms of factors related to their roles, job satisfaction and physical condition played in general a greater role for radiological technologists (p<0.001). Immediate seniors and behavioral conducts were also found to have a lot influence on them (p<0.01).3. As for job conflicts, the kind of medical institutions seemed to exercise a great deal of impact on radiological technologists (p<0.05), while colleagues, immediate seniors, job satisfaction and physical conditions also produced a lot of influence on them as a whole (p<0.001).4. Regarding job autonomy, radiological technologists were found to have been less influenced by position in metropolitan cities and by immediate seniors in the capital city (p<0.01). As a whole, however, age and job satisfaction acted as decisive factors of influence on them (p<0.001). 5. Among factors related to job burden, gender played a bigger role for radiological technologists in metropolitan cities as the kind of medical institutions did in small- and medium-sized cities. Centrally, job satisfaction and behavioral conducts exercised a great impact on them (p<0.001).6. Job stability seemed to be more immensely linked to gender and physical conditions in metropolitan cities whereas position did more in the capital city. In general, how ever, job satisfaction produced a far-reaching (p<0.001).