• Title/Summary/Keyword: Systems Biotechnology

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RNAseq-based Transcriptome Analysis of Burkholderia glumae Quorum Sensing

  • Kim, Sunyoung;Park, Jungwook;Kim, Ji Hyeon;Lee, Jongyun;Bang, Bongjun;Hwang, Ingyu;Seo, Young-Su
    • The Plant Pathology Journal
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    • v.29 no.3
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    • pp.249-259
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    • 2013
  • Burkholderia glumae causes rice grain rot and sheath rot by producing toxoflavin, the expression of which is regulated by quorum sensing (QS). The QS systems of B. glumae rely on N-octanoyl homoserine lactone, synthesized by TofI and its cognate receptor TofR, to activate the genes for toxoflavin biosynthesis and an IclR-type transcriptional regulator gene, qsmR. To understand genome-wide transcriptional profiling of QS signaling, we employed RNAseq of the wild-type B. glumae BGR1 with QS-defective mutant, BGS2 (BGR1 tofI::${\Omega}$) and QS-dependent transcriptional regulator mutant, BGS9 (BGR1 qsmR::${\Omega}$). A comparison of gene expression profiling among the wild-type BGR1 and the two mutants before and after QS onset as well as gene ontology (GO) enrichment analysis from differential expressed genes (DEGs) revealed that genes involved in motility were highly enriched in TofI-dependent DEGs, whereas genes for transport and DNA polymerase were highly enriched in QsmR-dependent DEGs. Further, a combination of pathways with these DEGs and phenotype analysis of mutants pointed to a couple of metabolic processes, which are dependent on QS in B. glumae, that were directly or indirectly related with bacterial motility. The consistency of observed bacterial phenotypes with GOs or metabolic pathways in QS-regulated genes implied that integration RNAseq with GO enrichment or pathways would be useful to study bacterial physiology and phenotypes.

Imidazole Antifungal Drugs Inhibit the Cell Proliferation and Invasion of Human Breast Cancer Cells

  • Bae, Sung Hun;Park, Ju Ho;Choi, Hyeon Gyeom;Kim, Hyesook;Kim, So Hee
    • Biomolecules & Therapeutics
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    • v.26 no.5
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    • pp.494-502
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    • 2018
  • Breast cancer is currently the most prevalent cancer in women, and its incidence increases every year. Azole antifungal drugs were recently found to have antitumor efficacy in several cancer types. They contain an imidazole (clotrimazole and ketoconazole) or a triazole (fluconazole and itraconazole) ring. Using human breast adenocarcinoma cells (MCF-7 and MDA-MB-231), we evaluated the effects of azole drugs on cell proliferation, apoptosis, cell cycle, migration, and invasion, and investigated the underlying mechanisms. Clotrimazole and ketoconazole inhibited the proliferation of both cell lines while fluconazole and itraconazole did not. In addition, clotrimazole and ketoconazole inhibited the motility of MDA-MB-231 cells and induced $G_1$-phase arrest in MCF-7 and MDA-MB-231 cells, as determined by cell cycle analysis and immunoblot data. Moreover, Transwell invasion and gelatin zymography assays revealed that clotrimazole and ketoconazole suppressed invasiveness through the inhibition of matrix metalloproteinase 9 in MDA-MB-231 cells, although no significant changes in invasiveness were observed in MCF-7 cells. There were no significant changes in any of the observed parameters with fluconazole or itraconazole treatment in either breast cancer cell line. Taken together, imidazole antifungal drugs showed strong antitumor activity in breast cancer cells through induction of apoptosis and $G_1$ arrest in both MCF-7 and MDA-MB-231 cells and suppression of invasiveness via matrix metalloproteinase 9 inhibition in MDA-MB-231 cells. Imidazole drugs have well-established pharmacokinetic profiles and known toxicity, which can make these generic drugs strong candidates for repositioning as antitumor therapies.

Comparative Investigation of Glutathione S-Transferases, Glyoxalase-I and Alliinase Activities in Different Vegetable Crops

  • Hossain, Md Daud;Rohman, Md Motiar;Fujita, Masayuki
    • Journal of Crop Science and Biotechnology
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    • v.10 no.1
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    • pp.19-26
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    • 2007
  • Glutathione S-transferases(GSTs, EC 2.5.1.18), glyoxalase-I(EC 4.4.1.5) and alliin lyase(alliinase, EC 4.4.1.4) are important enzyme systems in plant bodies. The first two are mainly detoxifying enzymes that utilize glutathione(GSH) in the defense mechanism, and the last one is mainly involved in secondary metabolism and relevant to sulfur compounds derived from GSH. The activities of the three enzymes have been investigated in soluble extracts of vegetable crops, including pumpkin, cabbage, broccoli, radish, carrot, potato, sweet potato, mungbean, and onion. GST activities were detected in all of the vegetables, and the extract of onion bulb exhibited the highest specific activity(648 nmol/min/mgP). The putative GSTs of most of the vegetables were found to be induced by ethanol. The activities of GSTs in onion bulb were found to be markedly inhibited by S-hexyl glutathione and were also inhibited by S-butyl glutathione and S-propyl glutathione. The anti-CmGSTF1 antiserum recognized a thick band for putative onion GST. The estimated glyoxalase-I activity level was also high in onion bulb(4540 nmol/min/mgP), indicating that the thick band detected by Western blot analysis might result from partial recognition of glyoxalase-I by the antiserum. The specific activities for glyoxalase-I were moderate in radish and carrot, and the extracts of other vegetables had rather low levels of activities. The extract of onion also showed the highest specific activity level for alliinase(2069nmol pyruvate/mgP). The extracts of other vegetables also had alliinase activities, although the estimated values were much lower than that of onion.

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Characteristic Changes in Brown Rice (Oryza sativa L.) Cultivars of 3 Ecotypes During Different Storage Conditions

  • Oh, Sea-Kwan;Hwang, Pil-Seong;Lee, Choon-Ki;Kim, Yeon-Gyu;Seo, Woo-Duck;Cho, Kye-Man;Choung, Myoung-Gun;Lee, Jin-Hwan
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1091-1095
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    • 2009
  • The aim of this study investigated the fluctuations of 3 characters from 3 ecotypes [early ripening (ER), middle ripening (MR), and late ripening (LR)] of 20 Korean brown rice cultivars in different storage systems [time: 12 and 24 weeks, temperature: low ($10^{\circ}C$) and room ($25^{\circ}C$)]. With increase of storage time and temperature, lipoxygenase activity, and fat acidity increased, whereas germination rate was reduced. ER cultivars exhibited the highest lipoxygenase activity of $35.49{\pm}2.46$ unit/mg protein during 24 weeks storage at $25^{\circ}C$, followed by LR ($32.73{\pm}1.33$) and MR ($32.66{\pm}1.62$) cultivars. The amounts of fat acidity also were observed by the same order (ER: $20.40{\pm}2.12$>LR: $19.68{\pm}1.86$>MR: $19.64{\pm}1.35$ mg KOH/100 g). Germination rate slightly decreased with increase of time and temperature (MR>LR>ER), but MR and LR cultivars showed the most significant changes (ER: $60.90{\pm}23.47%$, MR: $32.66{\pm}13.95%$, and LR: $32.53{\pm}5.87%$). On the basis of above results, MR cultivars were evaluated the highest quality, because high lipoxygenase activity, high fat acidity, and low germination rate have deteriorated in quality and generated off-odor. Thus, MR cultivars might be very important sources in food processing and stored dietary supplement aspects.

Simple and Sensitive Liquid Chromatography Electrospray Ionization Mass Spectrometry Method for Determination of Glycoalkaloids in Potato (Solanum tuberosum L.)

  • Kim, Jae-Kwang;Bae, Shin-Cheol;Baek, Hyung-Jin;Seo, Hyo-Won;Ryu, Tae-Hun;Kim, Jung-Bong;Won, So-Youn;Sohn, Soo-In;Kim, Dong-Hern;Kim, Sun-Ju;Cho, Myoung-Rae
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.113-117
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    • 2009
  • A method was developed using enhanced liquid chromatography coupled with electrospray ionization mass spectrometry for the analysis and quantitation of 2 main potato glycoalkaloids, $\alpha$-chaconine, and $\alpha$-solanine, without any pre-concentration or derivatisation steps. Calibration curves generated by this technique exhibited a linear dynamic range from 0.025 to $50{\mu}g/mL$ and from 0.05 to $50{\mu}g/mL$ for $\alpha$-chaconine and $\alpha$-solanine, respectively. Matrix effects were evaluated by comparing calibration curves measured in matrix-matched and solvent-based systems. Ion suppression due to matrix effects was weak and extraction recoveries of 88 to 114% were obtained in different sample matrices spiked with analyte concentrations ranging from 15 to $35{\mu}g/mL$. Potatoes that had been genetically modified to tolerate glufosinate contained the same glycoalkaloid levels as their non-transgenic counterpart. We suggest complementing compositional comparison assessment strategy by validating quantitative analytical methods for the toxic glycoalkaloids in potato plants.

Technical Reviews on Ecosystem Modeling Approach and its Applicability in Ecosystem-Based Coastal Management in Saemangeum Offshore and Geum River Estuary (생태계기반 연안관리를 위한 생태모델 개발방향에 대한 기술적 검토 - 새만금 외해역 및 금강 하구역 사례)

  • Kim, Hae-Cheol;Kim, Yong Hoon;Chang, Won-Keun;Ryu, Jongseong
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.18 no.3
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    • pp.233-244
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    • 2015
  • Marine ecosystem modelling has become a more widely used decision-making tool in coastal ecosystem-based management. However, it is not trivial to develop a well calibrated/validated model with potential applicability and practicality because understanding ecological processes with complexities is a pre-requisite for developing robust ecosystem models and this accompanies a great deal of well coordinated efforts among field-going ecologists, laboratory scientists, modelers, stake-holders and managers. This report aims to provide a brief introduction on two different approaches in marine ecological models: deterministic (mechanistic) and stochastic (statistical) approach. We also included definitions, historical overview of past researches, case studies, and contextual suggestions for coastal management in Korea. A long list of references this report included in this study might be used as an introductory material for those who wish to enter ecosystem modelling field.

Inducible spy Transcription Acts as a Sensor for Envelope Stress of Salmonella typhimurium

  • Jeong, Seon Mi;Lee, Hwa Jeong;Park, Yoon Mee;Kim, Jin Seok;Lee, Sang Dae;Bang, Iel Soo
    • Food Science of Animal Resources
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    • v.37 no.1
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    • pp.134-138
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    • 2017
  • Salmonella enterica infects a broad range of host animals, and zoonostic infection threatens both public health and the livestock and meat processing industries. Many antimicrobials have been developed to target Salmonella envelope that performs essential bacterial functions; however, there are very few analytical methods that can be used to validate the efficacy of these antimicrobials. In this study, to develop a potential biosensor for Salmonella envelope stress, we examined the transcription of the S. enterica serovar typhimurium spy gene, the ortholog of which in Escherichia coli encodes Spy (${\underline{s}}pheroplast$ ${\underline{p}}rotein$ ${\underline{y}}$). Spy is a chaperone protein expressed and localized in the periplasm of E. coli during spheroplast formation, or by exposure to protein denaturing conditions. spy expression in S. typhimurium was examined by constructing a spy-gfp transcriptional fusion. S. typhimurium spy transcription was strongly induced during spheroplast formation, and also when exposed to membrane-disrupting agents, including ethanol and the antimicrobial peptide polymyxin B. Moreover, spy induction required the activity of regulator proteins BaeR and CpxR, which are part of the major envelope stress response systems BaeS/BaeR and CpxA/CpxR, respectively. Results suggest that monitoring spy transcription may be useful to determine whether a molecule particularly cause envelope stress in Salmonella.

Effects of Hydrophilic Additives on the Release Rate of Protein Drugs (단백질 약물 방출속도에 미치는 친수성 첨가제의 영향)

  • Kwon, Young-Kwan;Kim, Ji-Hyeon;Yoo, Young-Je
    • KSBB Journal
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    • v.22 no.4
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    • pp.213-217
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    • 2007
  • It has been reported that hydrophobic additives generally decrease the release rate of protein drugs from drug delivery systems (DDS) and hydrophilic additives increase the release rate. In many cases, however, the addition of hydrophilic molecule is necessary for improving the stability of protein drugs. In the present work, the effects of hydrophilic additives on the release profiles, and micelle formation of protein drug formulations were investigated to develop a novel method for protein drug delivery. For model protein drug, bovine serum albumin (BSA) was employed and several hydrophilic additives were used in the release experiments. Hydrophilic additive D-sorbitol showed the lower release rates of BSA than other hydrophobic additives due to the gel strengthening ability of the additive and the optimum concentration of D-sorbitol was 3 w/v % for the retarded release rate. In addition, it was found that the addition of D-sorbitol was very effective for obtaining homogeneous and stable DDS. The results were discussed in terms of the micelle formation and the micelle structure, i.e., the differences in gel structure and the distribution of drugs in micelles.

Genetic Synthesis and Applications of Repetitive Protein Polymers (반복단위 단백질 고분자의 유전공학적 합성 및 응용)

  • Park, Mi-Sung;Choi, Cha-Yong;Won, Jong-In
    • KSBB Journal
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    • v.22 no.4
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    • pp.179-184
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    • 2007
  • This study introduces the characteristics and some applications of repetitive polypeptides, especially to the biomaterial, tissue engineering scaffolds, drug delivery system, and DNA separation systems. Since some fibrous proteins, which consist of repeating peptide monomers, have been reported that their physical properties are changed dramatically by means of temperature alteration or pH shifting. For that reason, fibrous protein-mimetic polypeptides, which are produced by the recombinant technology, can be applied to the diverse biological fields. Repetitive polypeptides can also be used in the bioseparation area such as DNA sequencing, because they make DNA separation possible in free-solution electrophoresis by conjugating DNA fragments to them. Moreover, artificial synthesis of repetitive polypeptides helps to demonstrate the correlations between mechanical properties and structures of natural protein polymer, which have been proven that repetitive domains are affected by the sequence of the repeating domains and the number of repeating subunits. Repetitive polypeptides can be biologically synthesized using some special cloning methods, which are represented here. Recursive directional ligation (RDL) and controlled cloning method (CCM) have been proposed as excellent cloning methods in that we can control the number of repetition in the multimerization of polypeptides and the components of repetitive polypeptides by either method.

Antioxidant Activity of a Chitin-degrading Bacterium Bacillus idriensis (CGH18) (키틴분해 박테리아 Bacillus idriensis (CGH18)의 항산화 활성)

  • Jung, Myoung Eun;Hong, Joo Wan;Lee, Jeong-Im;Kwak, Myoung Kuk;Kim, Hojun;Sohn, Jae Hak;Song, Young-Sun;Oh, Kwang-Suk;Seo, Youngwan
    • KSBB Journal
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    • v.28 no.4
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    • pp.217-224
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    • 2013
  • A bacterium CGH18 exhibiting antioxidizing and chitin-degrading activities in the colloidal chitin culture medium was isolated from salt-fermented crab. This strain was identified as Bacillus idriensis based on 16S rDNA sequence homology search. Its crude extract was partitioned between n-BuOH and $H_2O$. The organic layer was further partitioned between $CH_2$ $Cl_2$ and $H_2O$. Antioxidant activities of crude extract and its solvent fractions were evaluated using five different bioassay methods, including the degree of occurrence of intracellular reactive oxygen species (ROS), peroxynitrite scavenging (ONOO), and oxidative damage of genomic DNA. All fractions exhibited significant antioxidant activity in bioassay systems used.