• Title/Summary/Keyword: Systems Biology

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Treatment of Epidermal Growth Factor (EGF) enhances Nuclear Maturation of Porcine Oocytes and Stimulates Expression of ER/Golgi Transport Proteins

  • Hwangbo, Yong;Oh, Hae-In;Lee, Sang-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Development and Reproduction
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    • v.21 no.2
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    • pp.131-138
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    • 2017
  • This study was conducted to investigate stimulatory effect of epidermal growth factor (EGF) on nuclear maturation and the expression level of EGF-receptor (EGFR), GM-130 (a marker of Golgi apparatus), transport protein Sec61 subunit beta ($Sec61{\beta}$), and coatomer protein complex subunit gamma 2 (COPG2) in porcine oocytes. The cumulus-oocyte complexes were collected from follicle with 3-6 mm in diameter. They were incubated in medium with/without EGF for 22 h (IVM I) and subsequently incubated hormone-free medium with/without EGF for 22 h (IVM II). Nuclear maturation state was checked by aceto-orcein stain. Protein expression of EGFR, GM-130, $Sec61{\beta}$, and COPG2 were measured by immunofluorescence. In results, nuclear maturation of oocytes in EGF non-treated oocytes were significantly lower than EGF-treated groups at IVM I or IVM II stage (P<0.05), whereas maturational rate in EGF treatment groups at both of IVM stage was higher in among the all treatment groups (P<0.05). EGFR, GM-130, $Sec61{\beta}$ and COPG2 were expressed in the cytoplasm of oocytes. Especially, GM-130 and EGFR were strongly expressed, but $Sec61{\beta}$ and COPG2 were weakly expressed in cortical area of cytoplasm. The protein level of GM-130, $Sec61{\beta}$, and COPG2 were significantly higher in the EGF-treated groups (P<0.05). However EGFR was no difference between non EGF-treated groups and control. In conclusion, EGF plays an important role in the systems for oocyte maturation with endoplasmic reticulum and Golgi apparatus. In addition, the protein levels of $Sec61{\beta}$ and COPG2 could be changed by EGF in the porcine oocytes during maturation.

Comparison of Foliar Ultrastructure of 3 Dubautia species (Dubautia속(屬) 3종(種) 식물(植物)의 엽육조직(葉肉組織) 미세구조(微細構造) 비교연구(比較硏究))

  • Kim, In-Sun
    • Applied Microscopy
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    • v.24 no.4
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    • pp.13-31
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    • 1994
  • The fine structure of palisade chloroplasts has been studied in the mature leaves of 3 Dubautia species (D. scabra var. leiophylla, D. knudsenii and D. scabra var. leiophylla${\times}$D. knudsenii) to explore variation at the ultrastructural level, since the parental species exhibit quite different morphological and anatomical features. Types of thylakoidal membrane systems, occurrence and distribution of phytoferritin-like structures, lipid droplets, starch grains, mitochondria and microbodies were examined. Four different types of thylakoidal membranes were found in D. scabra var. leiophylla, 2 rather uniform types in D. knudsenii and 3 intermediate types in their hybrid. D. scabra var. leiophylla and the hybrid were marked by statistically significant differences in mean numbers of thylakoids per granum, while no significant difference was found between D. knudsenii and the hybrid. Phytoferritin-like structures which were about $100-120{\AA}$ in diameter as a whole particle each were found in all 3 species. The amount and distribution of particles varied by species. Lipid droplets, plastoglobuli, and starch grains occurred in all 3 species, but the frequency of starch grains also varied with the species. More frequent and larger starch grains were observed in D. knudsenii than in the other two species. Microbodies, or peroxisome, were observed throughout all species. They occurred, either with or without crystalline inclusions, around the chloroplasts.

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Gene annotation by the "interactome"analysis in KEGG

  • Kanehisa, Minoru
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2000.11a
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    • pp.56-58
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    • 2000
  • Post-genomics may be defined in different ways depending on how one views the challenges after the genome. A popular view is to follow the concept of the central dogma in molecular biology, namely from genome to transcriptome to proteome. Projects are going on to analyze gene expression profiles both at the mRNA and protein levels and to catalog protein 3D structure families, which will no doubt help the understanding of information in the genome. However complete, such catalogs of genes, RNAs, and proteins only tell us about the building blocks of life. They do not tell us much about the wiring (interaction) of building blocks, which is essential for uncovering systemic functional behaviors of the cell or the organism. Thus, an alternative view of post-genomics is to go up from the molecular level to the cellular level, and to understand, what I call, the "interactome"or a complete picture of molecular interactions in the cell. KEGG (http://www.genome.ad.jp/kegg/) is our attempt to computerize current knowledge on various cellular processes as a collection of "generalized"protein-protein interaction networks, to develop new graph-based algorithms for predicting such networks from the genome information, and to actually reconstruct the interactomes for all the completely sequenced genomes and some partial genomes. During the reconstruction process, it becomes readily apparent that certain pathways and molecular complexes are present or absent in each organism, indicating modular structures of the interactome. In addition, the reconstruction uncovers missing components in an otherwise complete pathway or complex, which may result from misannotation of the genome or misrepresentation of the KEGG pathway. When combined with additional experimental data on protein-protein interactions, such as by yeast two-hybrid systems, the reconstruction possibly uncovers unknown partners for a particular pathway or complex. Thus, the reconstruction is tightly coupled with the annotation of individual genes, which is maintained in the GENES database in KEGG. We are also trying to expand our literature surrey to include in the GENES database most up-to-date information about gene functions.

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The Activities of Antioxidant Enzymes in Response to Oxidative Stresses and Hormones in Paraquat-tolerant Rehmannia glutinosa Plants

  • Choi, Dong-Geun;Yoo, Nam-Hee;Yu, Chang-Yeon;De Los Reyes, Benildo;Yun, Song-Joong
    • BMB Reports
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    • v.37 no.5
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    • pp.618-624
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    • 2004
  • All members of R. glutinosa show the unique characteristic of intrinsic tolerance to paraquat (PQ). Antioxidant enzymes have been proposed to be the primary mechanism of PQ resistance in several plant species. Therefore, the antioxidant enzyme systems of R. glutinosa were evaluated by comparatively analyzing cellular antioxidant enzyme levels, and their responses of oxidative stresses and hormones. The levels of ascorbate peroxidase (APX), glutathione reductase (GR), non-specific peroxidase (POX), and superoxide dismutase (SOD) were 7.3-, 4.9-, 2.7- and 1.6-fold higher in PQ-tolerant R. glutinosa than in PQ-susceptible soybeans. However, the activity of catalase (CAT) was about 12-fold higher in the soybeans. The activities of antioxidant enzymes reduced after PQ treatment in the two species, with the exception of POX and SOD in R. glutinosa, which increased by about 40%. Interestingly, the activities of APX, SOD and POX in R. glutinosa, relative to those in soybeans, were further increased by 49, 67 and 93% after PQ treatment. The considerably higher intrinsic levels, and increases in the relative activities of antioxidant enzymes in R. glutinosa under oxidative stress support the possible role of these enzymes in the PQ tolerance of R. glutinosa. However, the relatively lower levels of SOD versus PQ tolerance, and the mixed responses of antioxidant enzymes to stresses and hormones, suggest a possible alternative mechanism(s) for PQ tolerance in R. glutinosa.

Staphylococcus aureus Siderophore-Mediated Iron-Acquisition System Plays a Dominant and Essential Role in the Utilization of Transferrin-Bound Iron

  • Park Ra Young;Sun Hui Yu;Choi Mi Hwa;Bai Young Hoon;Shin Sung Heui
    • Journal of Microbiology
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    • v.43 no.2
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    • pp.183-190
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    • 2005
  • Staphylococcus aureus is known to be capable of utilizing transferrin-bound iron, via both siderophore­and transferrin-binding protein (named IsdA)-mediated iron-acquisition systems. This study was designed in order to determine which iron-acquisition system plays the essential or dominant role with respect to the acquisition of iron from human transfenin, in the growth of S. aureus. Holotransferrin (HT) and partially iron-saturated transferrin (PT), but not apotransferrin (AT), were found to stimulate the growth of S. aureus. S. aureus consumed most of the transferrin-bound iron during the exponential growth phase. Extracellular proteases were not, however, involved in the liberation of iron from transferrin. Transferrin-binding to the washed whole cells via IsdA was not observed during the culture. The expression of IsdA was observed only in the deferrated media with AT, but not in the media supplemented with PT or HT. In contrast, siderophores were definitely produced in the deferrated media with PT and HT, as well as in the media supplemented with AT. The siderophores proved to have the ability to remove iron directly from transferrin, but the washed whole cells expressing IsdA did not. In the bioassay, the growth of S. aureus on transferrin-bound iron was stimulated by the siderophores alone. These results demonstrate that the siderophore-mediated iron-acquisition system plays a dominant and essential role in the uptake of iron from transferrin, whereas the IsdA-mediated iron-acquisition system may play only an ancillary role in the uptake of iron from transferrin.

Characterization of a Squalene Synthase from the Thraustochytrid Microalga Aurantiochytrium sp. KRS101

  • Hong, Won-Kyung;Heo, Sun-Yeon;Park, Hye-Mi;Kim, Chul Ho;Sohn, Jung-Hoon;Kondo, Akihiko;Seo, Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.759-765
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    • 2013
  • The gene encoding squalene synthase (SQS) of the lipid-producing heterotrophic microalga Aurantiochytrium sp. KRS101 was cloned and characterized. The krsSQS gene is 1,551 bp in length and has two exons and one intron. The open reading frame of the gene is 1,164 bp in length, yielding a polypeptide of 387 predicted amino acid residues with a molecular mass of 42.7 kDa. The deduced krsSQS sequence shares at least four conserved regions known to be required for SQS enzymatic activity in other species. The protein, tagged with $His_6$, was expressed into soluble form in Escherichia coli. The purified protein catalyzed the conversion of farnesyl diphosphate to squalene in the presence of NADPH and $Mg^{2+}$. This is the first report on the characterization of an SQS from a Thraustochytrid microalga.

Effects of Reactive Oxygen Species and Antioxidants during In Vitro Maturation Oocytes and Embryo Development in Pigs (돼지 난자의 체외성숙과 배아발달 동안 ROS와 항산화제의 영향)

  • Lee, Won-Hee;Park, Ji-Eun;Hwangbo, Yong;Kim, Hwa-Young;Lee, Ji-Eun;Kang, Byeong-Buhm;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.41 no.1
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    • pp.17-23
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    • 2017
  • The oocyte undergoes various events during in vitro maturation (IVM) and subsequence development. One of the events is production of reactive oxygen species (ROS) that is a normal process of cell metabolism. But imbalances between ROS production and antioxidant systems induce oxidative stress that negatively affect to mammalian reproductive process. In vitro environments, in vitro matured oocytes have many problems, such as excessive production of ROS and imperfect cytoplasmic maturation. Therefore, in vitro matured oocytes still have lower maturation rates and developmental competence than in vivo matured oocytes. In order to improve the IVM and in vitro culture (IVC) system, antioxidants, vitamins were added to the IVM, IVC medium. Antioxidant supplementation was effective in controlling the production of ROS and it continues to be explored as a potential strategy to overcome mammalian reproductive disorders. Based on these studies, we expect that the use of antioxidants in porcine oocytes could improved maturation and development rates.

Dependence Potential of Quetiapine: Behavioral Pharmacology in Rodents

  • Cha, Hye Jin;Lee, Hyun-A;Ahn, Joon-Ik;Jeon, Seol-Hee;Kim, Eun Jung;Jeong, Ho-Sang
    • Biomolecules & Therapeutics
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    • v.21 no.4
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    • pp.307-312
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    • 2013
  • Quetiapine is an atypical or second-generation antipsychotic agent and has been a subject of a series of case report and suggested to have the potential for misuse or abuse. However, it is not a controlled substance and is not generally considered addictive. In this study, we examined quetiapine's dependence potential and abuse liability through animal behavioral tests using rodents to study the mechanism of quetiapine. Molecular biology techniques were also used to find out the action mechanisms of the drug. In the animal behavioral tests, quetiapine did not show any positive effect on the experimental animals in the climbing, jumping, and conditioned place preference tests. However, in the head twitch and self-administration tests, the experimental animals showed significant positive responses. In addition, the action mechanism of quetiapine was found being related to dopamine and serotonin release. These results demonstrate that quetiapine affects the neurological systems related to abuse liability and has the potential to lead psychological dependence, as well.

Enhancing resistance to major fungal pathogens of Panax ginseng, by BTH-induced systemic resistance (BTH 처리한 배배양 인삼에서 주요 진균병 저항성 증진 효과)

  • Ryu, Hojin
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.99-103
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    • 2016
  • In perennial ginseng plantations, the effective control of various diseases is one of the most critical factors for increasing yields. Enhancing the resistance to disease through induced systemic resistance (ISR) and anti-microbial activity of beneficial soil bacteria, is currently considered to be a potential promising approach to integrate pathogen management for sustainable agriculture. However, the effective in vitro culture systems for testing ISR in ginseng plants have been rarely reported. In this study, I have successfully developed an in vitro germ-free culture system of Panax ginseng seedling for diverse purposes. With this useful system, we also tested BTH-induced priming effects against Botrytis cinerea and Colletotrichum panacicola. Compared to the drain method for enhancing ISR effects to ginseng seedlings, the direct method of spraying leaves somewhat increased the defense activity to these major fungal pathogens. Consistently, the expression of pathogen related PgPR10 and PgCAT were greatly and rapidly enhanced in the BTH-treated ginseng seedlings by treatment with C. panacicola. Our results revealed that the in vitro culture system can be used for developing eco-friendly and versatile bio-control agents for harmful diseases in ginseng cultivation.

Agricultural biotechnology: Opportunities and challenges associated with climate change (기후변화에 대응한 농업생명공학의 기회와 도전)

  • Chang, An-Cheol;Choi, Ji-Young;Lee, Shin-Woo;Kim, Dong-Hern;Bae, Shin-Chul
    • Journal of Plant Biotechnology
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    • v.38 no.2
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    • pp.117-124
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    • 2011
  • Considering that the world population is expected to total 9 billion by 2050, it will clearly be necessary to sustain and even accelerate the rate of improvement in crop productivity. In the 21st century, we now face another, perhaps more devastating, environmental threat, namely climate change, which could cause irreversible damage to agricultural ecosystem and loss of production potential. Enhancing intrinsic yield, plant abiotic stress tolerance, and pest and pathogen resistance through agricultural biotechnology will be a critical part of feeding, clothing, and providing energy for the human population, and overcoming climate change. Development and commercialization of genetically engineered crops have significantly contributed to increase of crop yield and farmer's income, decrease of environmental impact associated with herbicide and insecticide, and to reduction of greenhouse gas emissions from this cropping area. Advances in plant genomics, proteomics and system biology have offered an unprecedented opportunities to identify genes, pathways and networks that control agricultural important traits. Because such advances will provide further details and complete understanding of interaction of plant systems and environmental variables, biotechnology is likely to be the most prominent part of the next generation of successful agricultural industry. In this article, we review the prospects for modification of agricultural target traits by genetic engineering, including enhancement of photosynthesis, abiotic stress tolerance, and pest and pathogen resistance associated with such opportunities and challenges under climate change.