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Safety assessment of the AtCYP78A7 protein expressed in genetically modified rice tolerant to abiotic stress

  • Nam, Kyong-Hee;Kim, Do Young;Shin, Hee Jae;Pack, In-Soon;Park, Jung-Ho;Yoon, Won Kee;Kim, Ho Bang;Kim, Chang-Gi
    • Korean Journal of Agricultural Science
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    • v.45 no.2
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    • pp.248-257
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    • 2018
  • Overexpression of AtCYP78A7, a gene encoding a cytochrome P450 protein, has been reported to improve tolerance to drought stress in genetically modified (GM) rice (Oryza sativa L.). The aim of this study was to evaluate the potential allergenicity and acute oral toxicity of the AtCYP78A7 protein expressed in GM rice. Bioinformatics analysis of the amino acid sequence of AtCYP78A7 did not identify any similarities with any known allergens or toxins. It showed that no known allergen had more than a 35% amino acid sequence homology with the AtCYP78A7 protein over an 80 amino acid window or more than 8 consecutive identical amino acids. The gene encoding the AtCYP78A7 protein was cloned in the pGEX-4T-1 vector and expressed in E. coli. Then, the AtCYP78A7 protein was purified and analyzed for acute oral toxicity. The AtCYP78A7 protein was fed at a dose of 2,000 mg/kg body weight in mice, and the changes in mortalities, clinical findings, and body weight were monitored for 14 days after the dosing. Necropsy was carried out on day 14. The protein did not cause any adverse effects when it was orally administered to mice at 2000 mg/kg body weight. These results indicate that the AtCYP78A7 protein expressed in GM rice would not be a potential allergen or toxin.

Simple and Novel Assay of the Host-Guest Complexation of Homocysteine with Cucurbit[7]uril

  • Park, Se-Ho;Lee, Jae-Yeul;Cho, Hyun-Nam;Kim, Kyoung-Ran;Yang, Seun-Ah;Kim, Hee-Joon;Jhee, Kwang-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.29 no.1
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    • pp.114-126
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    • 2019
  • This paper introduces three ways to determine host-guest complexation of cucurbit[7]uril (CB[7]) with homocysteine (Hcy). After preincubating Hcy and cysteine (Cys) with CB[7], Ellman's reagent (DTNB) was used to detect Hcy and Cys. Only Cys reacted with DTNB and Hcy gave a retarded color change. This suggests that the -SH group of Hcy is buried inside CB[7]. Human cystathionine ${\gamma}-lyase$ (hCGL) decreased the level of Hcy degradation after preincubating Hcy and CB[7]. These results suggest that the amount of free Hcy available was decreased by the formation of a Hcy-CB[7] complex. The immunological signal of anti-Hcy monoclonal antibody was decreased significantly by preincubating CB[7] with Hcy. The ELISA results also show that ethanethiol group ($-CH_2CH_2SH$) of Hcy, which is an epitope of anti-Hcy monoclonal antibody, was blocked by the cavity in CB[7]. Overall, CB[7] can act as a host by binding selectively with Hcy, but not Cys. The calculated half-complexation formation concentration of CB[7] was 58.2 nmol using Ellman's protocol, 97.9 nmol using hCGL assay and 87.7 nmol using monoclonal antibody. The differing binding abilities of Hcy and Cys towards the CB[7] host may offer a simple and useful method for determining the Hcy concentration in plasma or serum.

A study of the metabolites for 7-keto-DHEA-acetate in human urine (II) (뇨시료에 함유된 7-keto-DHEA-acetate의 대사체에 관한 연구 (II))

  • Kim, Yunje;Lee, Jinhee
    • Analytical Science and Technology
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    • v.17 no.5
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    • pp.401-409
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    • 2004
  • The 7-keto-DHEA-acetate is converted to 7-keto-DHEA, a metabolite of DHEA, and similar to its metabolism. We studied the metabolite M3, M4, and M5 of 7-keto-DHEA-acetate. The estimated molecular weight of M3 and M4 was 304 which were supposed to have more 3 hydroxyl and/or ketone groups. We could know that M3 is the 7-OH-DHEA which has the hydroxyl groups on 3 and 7-carbon and a ketone group on 17-carbon. In case of M4, it is the 7-oxo-diol metabolite which has the hydroxyl groups on 3 and 17-carbon and a ketone group on 7-carbon. The M5 was supposed that the molecular weight is 320 and has the three hydroxyl groups on 3, 6, and 16 carbon and the ketone group on 17-carbon. After dosing, 7-OH-DHEA showed the maximum urine flow in human urine after 5 hr and decreased rapidly. But we could find it until 58 hr why is a higher remaining substance.

Anti-inflammatory Effects of Haliotis discus-derived Antibacterial Peptide (AMP) Ab4-7 as a Functional material for Improving Inflammatory Skin Diseases (염증성 피부질환 개선 기능 소재로서 둥근전복(Haliotis discus) 유래 항균펩타이드 Ab4-7의 항염증 효과)

  • Choi, Soo-Cheol;Seo, Jung-Kil;Hwang, Joon-Ho;Lee, Ki-Young;Lee, In-Ah
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.2
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    • pp.147-157
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    • 2020
  • Abalone, a marine organism inhabiting the west coast of Korea, has antioxidant and anti-inflammatory effects, and is a resource with potential to be used in various industries such as antibiotic development and cosmetic raw materials. In this study, we chose abalone among various marine lives on the west coast. Antibacterial peptide (AMP) was separated from abalone and its derivative Ab4-7 was identified and its physiological activity was studied. The treatment of Ab4-7 in inflammatory RAW 264.7 to check the anti-inflammatory efficacy nhibited inflammatory cytokines, TLR4, TNF-α, IL-1β, COX-2, and iNOS and increased mRNA manifestation of HO-1, genes related to antioxidants. Based on the strong antioxidant and anti-inflammatory effects of Ab4-7, the effects of Ab4-7 in the inflammatory RAW 264.7 cells were identified through RT-PCR, which regulates the gene Matrix Metalloproteinase (MMPs) that induces a variety of inflammatory skin diseases by engaging in the decomposition of the extrocellular matrix metalloproteinase (ECM). Taken together, it is concluded that Ab4-7 has a powerful anti-inflammatory and antioxidant effect and can be used as a functional material for various inflammatory skin disease treatments by controlling the genes associated with matrix metalloproteinase (MMPs).

A Study on the Analysis of 1.7GHz Frequency Interference for Domestic Digital Cordless Phone (1.7GHz 국내 디지털 코드리스폰 도입을 위한 주파수 간섭 분석에 관한 연구)

  • Kang, Gun-Hwan;Kim, Jong-Ho;Park, Duk-Kyu
    • Proceedings of the Korea Contents Association Conference
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    • 2006.11a
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    • pp.52-57
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    • 2006
  • This research studies and analyzes the current trends and the frequency allocation bands for digital cordless phone(DCP) in other county. From these results, we propose 1.7GHz & 2.4GHz as a effective candidate frequency band for domestic DCP. A proposed 1.7GHz is expected to introduce DECT system of Europe. Therefore it is necessary to make an analysis of interference between 1.7GHz band and an adjacent IMT-2000 band. In this paper, we proposed the allocation of channel for 1.7GHz on the basis of the analysis of frequency interference between 1.7GHz band and an adjacent IMT-2000 band.

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Clustering of MPEG-7 Data for Efficient Management (MPEG-7 데이터의 효율적인 관리를 위한 클러스터링 방법)

  • Ahn, Byeong-Tae;Kang, Byeong-Shoo;Diao, Jianhua;Kang, Hyun-Syug
    • Journal of Korea Multimedia Society
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    • v.10 no.1
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    • pp.1-12
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    • 2007
  • To use multimedia data in restricted resources of mobile environment, any management method of MPEG-7 documents is needed. At this time, some XML clustering methods can be used. But, to improve the performance efficiency better, a new clustering method which uses the characteristics of MPEG-7 documents is needed. A new clustering improved query processing speed at multimedia search and it possible document storage about various application suitably. In this paper, we suggest a new clustering method of MPEG-7 documents for effective management in multimedia data of large capacity, which uses some semantic relationships among elements of MPEG-7 documents. And also we compared it to the existed clustering methods.

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Synthesis and in vitro Antitumor Activity of 2-Alkyl, 2-Aryl, and 2-Piperazinyl Benzimidazole-4, 7-dione Derivatives

  • Ahn, Chan-Mug;Tak, Jung-Ae;Choi, Sun-Ju
    • Archives of Pharmacal Research
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    • v.23 no.4
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    • pp.288-301
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    • 2000
  • A series of 2-alkyl, 2-aryl, and 2-piperazinyl benzimidazole-4,7-dione derivatives (7a-h) and 16m-o) were prepared, and their cytotoxicities were tested against three cancer cell lines (mouse lymphocytic leukemia cell line P388, and human gastric carcinoma cell lines SNU-1 and SNU-16). These compounds showed potent cytotoxicity against all of three cell lines tested, and especially SNU-16 was sensitive to them. 2-Aryl (7g,h) and 2-piperazinyl benzimidazole-4,7-dione derivative (I6 m) were more potent than mitomycin C against P388 and SNU-16. Among benzimidazole-4,7-dione derivatives with alkyl group at position 2, 7a had the most potent cytotoxicity against all of the cell lines tested.

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Cloning and Characterization of a Novel Laccase Gene, fvlac7, Based on the Genomic Sequence of Flammulina velutipes

  • Kim, Jong-Kun;Lim, Seon-Hwa;Kang, Hee-Wan
    • Mycobiology
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    • v.41 no.1
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    • pp.37-41
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    • 2013
  • Laccases (EC 1.10.3.2) are copper-containing polyphenol oxidases found in white-rot fungi. Here, we report the cloning and analysis of the nucleotide sequence of a new laccase gene, fvlac7, based on the genomic sequence of Flammulina velutipes. A primer set was designed from the putative mRNA that was aligned to the genomic DNA of F. velutipes. A cDNA fragment approximately 1.6-kb long was then amplified by reverse transcriptase-PCR using total RNA, which was subsequently cloned and sequenced. The cDNA sequence of fvlac7 was then compared to that of the genomic DNA, and 16 introns were found in the genomic DNA sequence. The fvlac7 protein, which consists of 538 amino acids, showed only 42~51% identity with 12 different mushroom species containing two laccases of F. velutipes, suggesting the fvlac7 is a novel laccase gene. The first 25 amino acids of Fvlac7 correspond to a predicted signal sequence, four copper-binding sites, and four N-glycosylation sites. Fvlac7 cDNA was heterologously overexpressed in an Escherichia coli system with an approximate expected molecular weight of 60 kDa.

Molecular Cloning and Characterization of Two Major Endoglucanases from Penicillium decumbens

  • Wei, Xiao-Min;Qin, Yu-Qi;Qu, Yin-Bo
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.265-270
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    • 2010
  • Two major endoglucanase genes (cel7B and cel5A) were cloned from Penicillium decumbens 114-2 using the method of modified thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR). The result of Southern blotting suggested that P. decumbens has a single copy of the cel5A gene and a single copy of the cel7B gene in its chromosomal DNA. The expression levels of cel5A and cel7B were determined by means of real-time quantitative PCR, suggesting that the two genes were coordinately expressed, and repressed by glucose and induced by cellulose. Both endoglucanase genes were expressed in Saccharomyces cerevisiae and the recombinant proteins were purified. The recombinant Cel7B and Cel5A were both optimally active at $60^{\circ}C$ and pH 4.0. The recombinant Cel7B showed more than 8-fold, 30-fold, and 5-fold higher enzyme activities toward carboxymethyl cellulose, barley $\beta$-glucan, and PASC, respectively, in comparison with that of Cel5A. However, their activities toward pNPC and Avicel showed minor differences. The results suggested that Cel7B is a strict endoglucanase, whereas Cel5A showed processivity because of its relative higher ability to hydrolyze the crystal cellulose.

Effect of Duration Treated on Reproductive Performance in Holstein Cows Treated with Progesterone with and without Estradiol (인공수정 후 외인성 Progesterone과 Estradiol 병용 및 Progesterone 단독 투여 기간이 젖소의 번식 성적에 미치는 영향)

  • Baek, K.S.;Lee, W.S.;Park, S.B.;Park, S.J.;Kim, H.S.;Lee, H.J.;Ki, K.S.;Kang, S.J.;Ahn, B.S.
    • Journal of Embryo Transfer
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    • v.22 no.3
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    • pp.195-198
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    • 2007
  • This study was carried out to investigate the effects of hormones treated (progesterone, and combination of progesterone and estradiol) on the reproductive performance of dairy cattle. The intravaginal CIDR was inserted in the vagina of cow on day 14 (P-7, PE-7), 16 (P-5, PE-5) and 18 (P-3, PE-3) post-insemination to stimulate progesterone secretion with (PE) and without estradiol(P). The CIDR was removed on 7 (P-7, PE-7), 5 (P-5, PE-5) and 3 (P-3, PE-3) days after. The cows for control group were not treated with CIDR or hormones. Conception rate, the duration needed for conception after AI and the number of AI services needed for conception were measured. Conception rate in control, CIDR without estradiol (P-7, P-5 and P-3) and CIDR treated (PE-7, PE-5 and PE-3) cows were 15.8%, 40.0%, 31.5%, 28.6%, 62.5%, 50.0% and 0%, respectively. The days needed for conception after AI in control, P-7, P-5, p-3, PE-7, PE-5 and PE-3 were 64.9, 63.0, 59.1, 8.0, 0.0, 18.9 and 83.3 days, respectively. The days needed for conception in cows treated with CIDR (PE-7) has shown longer than control and PE-3 (p<0.05). The number of AI services needed for conception in control, CIDR (PE-7, PE-5 and PE-3) and CIDR without estradiol treated cows (P-7, P-5 and P-3) were 27, 2.3, 1.9, 1.3, 1.0, 1.7 and 2.8 times, respectively. The number of AI services needed for conception have shown significantly lower in PE-7 compared to control and PE-3.