• Title/Summary/Keyword: cell library

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Undecanoic Acid, Lauric Acid, and N-Tridecanoic Acid Inhibit Escherichia coli Persistence and Biofilm Formation

  • Jin, Xing;Zhou, Jiacheng;Richey, Gabriella;Wang, Mengya;Choi Hong, Sung Min;Hong, Seok Hoon
    • Journal of Microbiology and Biotechnology
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    • v.31 no.1
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    • pp.130-136
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    • 2021
  • Persister cell formation and biofilms of pathogens are extensively involved in the development of chronic infectious diseases. Eradicating persister cells is challenging, owing to their tolerance to conventional antibiotics, which cannot kill cells in a metabolically dormant state. A high frequency of persisters in biofilms makes inactivating biofilm cells more difficult, because the biofilm matrix inhibits antibiotic penetration. Fatty acids may be promising candidates as antipersister or antibiofilm agents, because some fatty acids exhibit antimicrobial effects. We previously reported that fatty acid ethyl esters effectively inhibit Escherichia coli persister formation by regulating an antitoxin. In this study, we screened a fatty acid library consisting of 65 different fatty acid molecules for altered persister formation. We found that undecanoic acid, lauric acid, and N-tridecanoic acid inhibited E. coli BW25113 persister cell formation by 25-, 58-, and 44-fold, respectively. Similarly, these fatty acids repressed persisters of enterohemorrhagic E. coli EDL933. These fatty acids were all medium-chain saturated forms. Furthermore, the fatty acids repressed Enterohemorrhagic E. coli (EHEC) biofilm formation (for example, by 8-fold for lauric acid) without having antimicrobial activity. This study demonstrates that medium-chain saturated fatty acids can serve as antipersister and antibiofilm agents that may be applied to treat bacterial infections.

A Study on the Performance Variation of CNTFET SRAM by the Partial Density Change of Carbon Nanotubes (탄소나노튜브 부분 밀도 변화에 의한 CNTFET SRAM 성능 변화에 대한 연구)

  • Cho, Geunho
    • Journal of IKEEE
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    • v.26 no.1
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    • pp.83-88
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    • 2022
  • With high performance and wide application, a CNTFET has been attracting a lot of attention as a next-eneration semiconductor, but the manufacturing process of CNTFET has not been mature enough, which makes commercialization difficult. In order to overcome the imperfections of the CNTFET manufacturing process and to increase the possibility of commercialization, this paper analyzes the CNTFET SRAM performance variation according to the CNTFET partial density change based on the recently reported CNTFET manufacturing process. Through HSPICE circuit simulation analysis using the existing 32nm CNTFET HSPICE library file, transistors whose performance variation is less sensitive to partial CNT density are selected among the six transistors constituting the SRAM cell and acceptable CNT density range is proposed. As the result of analysis, it is found that when the CNT density of the two transistors connected to the bit line in SRAM cell changed from 6/32nm to 8/32nm, the deviation of SRAM performance is less than 9% and when the CNT density is less than 5/32nm, the SRAM delay is increased by more than 8 time.

Novel Anti-Mesothelin Nanobodies and Recombinant Immunotoxins with Pseudomonas Exotoxin Catalytic Domain for Cancer Therapeutics

  • Minh Quan Nguyen;Do Hyung Kim;Hye Ji Shim;Huynh Kim Khanh Ta;Thi Luong Vu;Thi Kieu Oanh Nguyen;Jung Chae Lim;Han Choe
    • Molecules and Cells
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    • v.46 no.12
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    • pp.764-777
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    • 2023
  • Recombinant immunotoxins (RITs) are fusion proteins consisting of a targeting domain linked to a toxin, offering a highly specific therapeutic strategy for cancer treatment. In this study, we engineered and characterized RITs aimed at mesothelin, a cell surface glycoprotein overexpressed in various malignancies. Through an extensive screening of a large nanobody library, four mesothelin-specific nanobodies were selected and genetically fused to a truncated Pseudomonas exotoxin (PE24B). Various optimizations, including the incorporation of furin cleavage sites, maltose-binding protein tags, and tobacco etch virus protease cleavage sites, were implemented to improve protein expression, solubility, and purification. The RITs were successfully overexpressed in Escherichia coli, achieving high solubility and purity post-purification. In vitro cytotoxicity assays on gastric carcinoma cell lines NCI-N87 and AGS revealed that Meso(Nb2)-PE24B demonstrated the highest cytotoxic efficacy, warranting further characterization. This RIT also displayed selective binding to human and monkey mesothelins but not to mouse mesothelin. The competitive binding assays between different RIT constructs revealed significant alterations in IC50 values, emphasizing the importance of nanobody specificity. Finally, a modification in the endoplasmic reticulum retention signal at the C-terminus further augmented its cytotoxic activity. Our findings offer valuable insights into the design and optimization of RITs, showcasing the potential of Meso(Nb2)-PE24B as a promising therapeutic candidate for targeted cancer treatment.

An Efficient Hardware Implementation of ARIA Block Cipher Algorithm (블록암호 알고리듬 ARIA의 효율적인 하드웨어 구현)

  • Kim, Dong-Hyeon;Shin, Kyung-Wook
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2012.05a
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    • pp.91-94
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    • 2012
  • This paper describes an efficient implementation of ARIA crypto algorithm which is a KS (Korea Standards) block cipher algorithm. The ARIA crypto-processor supports three master key lengths of 128/192/256-bit specified in the standard. To reduce hardware complexity, a hardware sharing is employed, which shares round function in encryption/decryption module with key initialization module. It reduces about 20% of gate counts when compared with straightforward implementation. The ARIA crypto-processor is verified by FPGA implementation, and synthesized with a 0.13-${\mu}m$ CMOS cell library. It has 33,218 gates and the estimated throughput is about 640 Mbps at 100 MHz.

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Design and FPGA Implementation of Scalar Multiplication for A CryptoProcessor based on ECC(Elliptic Curve Cryptographics) (ECC(Elliptic Curve Crptographics) 기반의 암호프로세서를 위한 스칼라 곱셈기의 FPGA 구현)

  • Hwang Jeong-Tae;Kim Young-Chul
    • Proceedings of the IEEK Conference
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    • 2004.06b
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    • pp.529-532
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    • 2004
  • The ECC(Elliptic Curve Cryptogrphics), one of the representative Public Key encryption algorithms, is used in Digital Signature, Encryption, Decryption and Key exchange etc. The key operation of an Elliptic curve cryptosystem is a scalar multiplication, hence the design of a scalar multiplier is the core of this paper. Although an Integer operation is computed in infinite field, the scalar multiplication is computed in finite field through adding points on Elliptic curve. In this paper, we implemented scalar multiplier in Elliptic curve based on the finite field GF($2^{163}$). And we verified it on the Embedded digital system using Xilinx FPGA connected to an EISC MCU. If my design is made as a chip, the performance of scalar multiplier applied to Samsung $0.35 {\mu}m$ Phantom Cell Library is expected to process at the rate of 8kbps and satisfy to make up an encryption processor for the Embedded digital doorphone.

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Design of Efficient Flicker Detector for CMOS Image Sensor (CMOS Image sensor 를 위한 효과적인 플리커 검출기 설계)

  • Lee, Pyeong-Woo;Lee, Jeong-Guk;Kim, Chae-Sung
    • Proceedings of the IEEK Conference
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    • 2005.11a
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    • pp.739-742
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    • 2005
  • In this paper, an efficient detection algorithm for the flicker, which is caused by mismatching between light frequency and exposure time at CMOS image sensor (CIS), is proposed. The flicker detection can be implemented by specific hardware or complex signal processing logic. However it is difficult to implement on single chip image sensor, which has pixel, CDS, ADC, and ISP on a die, because of limited die area. Thus for the flicker detection, the simple algorithm and high accuracy should be achieved on single chip image sensor,. To satisfy these purposes, the proposed algorithm organizes only simple operation, which calculates the subtraction of horizontal luminance mean between continuous two frames. This algorithm was verified with MATLAB and Xilinx FPGA, and it is implemented with Magnachip 0.18 standard cell library. As a result, the accuracy is 95% in average on FPGA emulation and the consumed gate count is about 7,500 gates (@40MHz) for implementation using Magnachip 0.18 process.

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Molecular characterization and functional analysis of a protease-related protein in Chang-liver cells

  • Wang, Congrui;Zhang, Huiyong;Feng, Huigen;Yang, Baosheng;Pramanik, Jogenananda;Guo, Zhikun;Lin, Juntang
    • BMB Reports
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    • v.43 no.5
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    • pp.375-381
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    • 2010
  • In this study, the cDNA library of Chang-liver cells was immunoscreened using common ADAMs antibody to obtain ADAM related genes. We found one positive clone that was confirmed as a new gene by Blast, which is an uncharacterized helical and coil protein and processes protease activity, and named protease-related protein 1 (ARP1). The submitted GenBank accession number is AY078070. Molecular characterizations of ARP1 were analyzed with appropriate bioinformatics software. To analyse its expression and function, ARP1 was subcloned into glutathione S-transferase fusion plasmid pGEX-2T and expressed by E. coli system. The in vitro expression product of ARP1 was recognized by common ADAMs antibody with western blot. Interestingly, ARP1 cleaves gelatine at pH9.5, which suggests it is an alkaline protease. Semi-quantitative RT-PCR result indicates that ARP1 mRNA is strongly transcribed in the liver and the treated Chang-liver cells.

Production of Monoclonal Antibody about Specific Key Enzyme of Hyoscyamine $6{\beta}-Hydroxylase$ (H6H) in Scopolia parviflora

  • Kang, Young-Min;Jung, Hee-Young;Kang, Seung-Mi;Jin, Byung-Rae;Lee, Sang-Chul;Lee, Byung-Hyun;Choi, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.2
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    • pp.135-140
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    • 2004
  • Total RNAs were isolated from cultured roots of Scopolia parviflora, $poly(A)^+$ RNA was obtained through the mRNA purification, cDNA library of Hnh6h was constructed. Recombinant baculoviruses in Spodoptera frugiperda (Sf) cells were constructed by use of the transfer vector pBacPAK, which has the AcNPV sequence under the polyhedrin promoter. The expression vector carrying Hnh6h gene was transferred to S. parviflora and obtained transgenic hairy root lines. Our results confirmed the over expression of the H6H protein was used by anti-pBacPAK about cDNAs of S. parviflora. This study will served for production of tropane alkaloids by metabolic engineering.

Molecular cloning, tissue distribution and quantitative analysis of two proopiomelanocortin mRNAs in Japanese flounder (Paralichthys olivaceus)

  • Kim, Kyoung-Sun;Kim, Hyun-Woo;Chen, Thomas T.;Kim, Young-Tae
    • BMB Reports
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    • v.42 no.4
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    • pp.206-211
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    • 2009
  • Proopiomelanocortin (POMC) plays an essential role in the stress response of the hypothalamic-pituitary-adrenal axis, and is the precursor of biologically active peptides such as adrenocorticotropin (ACTH), $\alpha$-melanocyte-stimulating hormone ($\alpha$-MSH), $\beta$-melanocyte-stimulation hormone ($\beta$-MSH) and $\beta$-endorphin. We have synthesized two different forms of POMC cDNA clones, POMC-I and POMC-II, from a pituitary cDNA library for Paralichthys olivaceus, or Japanese flounder. jfPOMC-I cDNA consists of 954bp and encodes a polypeptide of 216 amino acid residues, whereas jfPOMC-II consists of 971bp which encode a polypeptide of 194 amino acid residues. The high levels of jfPOMC-I and -II mRNAs detected in the pituitary tissue and moderate levels detected in the brain tissue plus our quantitative RT-PCR analysis, which showed there to be no significant difference between the levels of jfPOMC-I and -II mRNAs, indicate that there may be no functional separation between these two mRNAs in the flounder.

cDNA Cloning and Overexpression of an Isoperoxidase Gene from Korean-Radish, Raphanus sativus L.

  • Park, Jong-Hoon;Kim, Soung-Soo
    • BMB Reports
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    • v.29 no.2
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    • pp.137-141
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    • 1996
  • A partial cDNA encoding a Korean radish isoperoxidase was obtained from a cDNA library prepared from 9 day old radish root. In order to obtain Korean radish isoperoxidase cDNA, 5' RACE (rapid amplification cDNA end) PCR was performed and a cDNA (prxK1) encoding a complete structural protein was obtained by RT (reverse transcription)-PCR. Sequence analysis revealed that the length of the cDNA was 945 base pairs, and that of the mRNA transcript was ca. 1.6 kb. The deduced amino acid of the protein were composed of 315 amino acid residues and the protein was 92% homologous to turnip peroxidase, and 46% to 50% homologous to other known peroxidases. The 945 bp cDNA encoding Korean radish isoperoxidase was overexpressed in Escherichia coli up to approximately 9% of total cellular protein. The recombinant fusion protein exhibited 43 kDa on SDS-PAGE analysis and the activity level of the recombinant nonglycosylated protein was two fold higher in IPTG induced cell extracts than that of uninduced ones.

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