Melandrin의 방향족 유도체 합성에 관한 연구

임중기,이강노,함원훈,이상인 성균관대학교 약학연구소 1992 成均藥硏論文集 Vol.4 No.1

Melandrin, as a possible phytoalexin, which is expected to have the various pharmacological activities; antibacterial, antifungal and antiviral activity, was isolated in 1987 by Woo from Melandrin firmum (Caryophyllaceae). The synthesis of melandrin derivatives has been accomplished by the reaction of 5-hydroxy anthranilic acid ethyl ester with acid chlorides and then by hydrolysis of ethyl ester.

효소면역측정법에 의한 우유중의 Aflatoxin M1 분석

손동화,임선희,이인원 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.5

국내산 우유중 aflatoxin M_1 (AFM_1)의 오염현황을 조사하기 위하여, 효소면역측정법(ELISA)을 개발하고 이를 이용한 독소의 정량을 행하였다. 항체의 생산을 위하여 AFM_1 bovine serum albumin 결합체를 Freund's adjuvant와 함께 토끼에 피하주사하여 면역하였다. 가장 높은 항체역가를 보인 항혈청과 AFB_1-horseradish peroxidase 결합체를 이용하여 직접법에 의한 경합적 효소면역측정법(competitive direct ELISA ; cdELISA)의 조건을 확립하였으며, 그 검출한계는 0.003ppb로 나타났다. 또한 유사독소에 대한 교차반응율은 aflatoxin M_1, M_2, B_1, _2, G_1, G_2, B_2a, and G_2a 에 대하여 각각 100, 29.9, 25.0, 2.7, 13.0, 0.65, 0, 및 0%이었다. 다음으로, AFM_1을 인위적으로 오염시킨 우유를 C_18 cartridge로 세척후 cdELISA로 분석하였을 때, 0.01~1 ppb(10~1,000 ppt)의 범위에서 그 회수율의 평균은 104%(CV의 평균, 6.4%)로 나타났다. 시유 및 목장우유를 대상으로 AFM_1의 오염현황을 cdELISA로 조사하였을 때, 평균 오염도는 80.4±55.0 ppt (n=64; range, 5,6~280 ppt)이었다. For a survey of the occurrence of aflatoxin M_1 (AFM_1) in domestic cow's milk, we developed an enzyme-linked immunosorbent assay (ELISA) system, and quantitated the toxin in cow's milk. In order to produce specific antibodies AFM_1 conjugated to bovine serum albumin (AFM_1-BSA) and Freund's adjuvant were immunized subcutaneously to rabbits. By use of the antiserum showing the highest titer and AFB_1-HRP conjugate, we established a competitive direct ELISA (cdELISA) for AFM_1 whose detection limit was 0.003 ppb. The cross-reactivities of the antiserum against aflatoxin M_1, M_2, B_1, _2, G_1, G_2, B_2a, and G_2a were 100, 29.9, 25.0, 2.7, 13.0, 0.65, 0, and 0%, respectively. When the cdELISA was applied to the cow's milk spiked with AFM_1 and followed by cleanup with C_18 cartridge, the mean recovery of the assay was 104% (mean of CV, 6.4%) in the final concentration of 0.01~1 ppb (10~1,000 ppt). When cow's milk samples gathered from markets and farms were assayed by the cdELISA, the mean concentration and SD of AFM_1 was 80.4±55.0 ppt (n=64; range, 5,6~280 ppt).

효소면역측정법을 이용한 Fumonisin의 검출법 개발

손동화,한성민,임선희,이인원,조선희,강신영,이경애 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1

Fumonisin에 대한 효소면역측정법(ELISA)을 개발하기 위하여 특이항체를 생산하고, 분석조건을 확립하여, 인위적으로 오염시킨 옥수수 시료의 분석을 행하였다. Fumonisin B_1 (FB_1)을 cholera toxin (CT)에 결합시킨 FB_1-CT conjugate를 면역원으로 하여 Freund's adjuvant와 함께, 또는 단독으로 두 군의 토끼에 면역하고 특이항체를 생산하였다. 항혈청을 ELISA로 분석한 결과 그 adjuvant와 함께 면역한 경우 항체역가(titer)가 높게 나타났다. 가장 높은 항체역가(1 : 16,000)를 나타낸 항혈청 및 그로부터 정제한 항체를 이용하여 간접법 및 직접법에 의한 경합적 ELISA(ciELISA 및 cdELISA)를 각각 확립하였다. 이 항체의 유사독소와의 교차반응을 ciELISA로 조사하였을 때, FB_3에 대한 교차반응율은 2%로 매우 낮았으나, FB_2에 대한 것은 179%로 다소 높았다. FB_1의 검출한계는 ciELISA에서 0.03ppb, cdELISA에서 0.3 ppb로 각각 나타났다. 실제 곡물시료의 분석에 ELISA 활용가능성을 조사하기 위하여 인위적으로 FB_1을 오염시킨 옥수수를 75% methanol로 추출하고 그 회수율을 측정한 결과, 분석이 불안정하여 회수율이 일정하게 나타나지 않았다. 그러나 방해물질의 제거를 위하여 시료추출액을 strong anion exchange(SAX) cartridge를 거쳐 세척한 다음 분석하였을 때, 3~10 ppm 범위의 FB_1 오염 옥수수에서 평균 34%(CV의 평균, 8.2%)의 안정된 회수율을 보였다. In order to develop enzyme-linked immunosorbent assay (ELISA) for fumonisins, production of specific antibodies, establishement of ELISA conditions, and quantitation of the toxin from spiked corns by ELISA were performed. Fumonisin B_1 (FB_1) conjugated to cholera toxin (CT) with or without Freund's adjuvant was subcutaneously injected into 2 groups of rabbits. When the titer of the antisera produced by each rabbit tested, higher titer was observed in case of the immunization with the adjuvant. By use of the antiserum showing the highest titer (1 : 16,000) and its purified antibodies, competitive indirect and direct ELISA's (ciELISA and cdELISA) were established, respectively. When the cross-reactivity of the antibody against fumonisin analogs was investigate by the ciELISA is was very low against B_3 (2%) but high against fumonisin B_2 (179%). The sensitivity of the ELISAs was also very high, because the detection limit for FB_1 was 0.03 ppb in ciELISA and 0.3 ppb in cdELISA. When the ELISA's were applied to the spiked corns after extraction with 75% methanol, the assay recovery of FB_1 was too unstable to assay. However, when cleanup by strong anion exchange (SAX) cartridge was introduced to remove interfering materials, the mean ELISA recovery of FB_1 from corns spiked to 3~10 ppm was found to be 34.0% and stable (mean of CV, 8.2%).

Targeted cellular delivery of robust enzyme nanoparticles for the treatment of drug-induced hepatotoxicity and liver injury

Lee, Min Sang,Kim, Nak Won,Lee, Jung Eun,Kim, Myung Goo,Yin, Yue,Kim, Sun Young,Ko, Bo Sung,Kim, Aeseon,Lee, Jong Han,Lim, Su Yeon,Lim, Dong Woo,Kim, Sun Hwa,Park, Ji Won,Lim, Yong Taik,Jeong, Ji Hoon Elsevier 2018 Acta Biomaterialia: structure-property-function re Vol.81 No.-

<P><B>Abstract</B></P> <P>Direct delivery of proteins into cells has been considered an effective approach for treating the protein-related diseases. However, clinical use of proteins has still been limited due to their instability in the blood and poor membrane permeability. To achieve an efficient cellular delivery of the protein to target cells via a systemic administration, a multifunctional carrier system having desirable stability both in the blood stream and the cells, specific cell-targeting property and endosomal escape functions may be required. In this study, we prepared a catalytic nanoparticle containing an active enzyme by cross-tethering multiple superoxide dismutase (SOD) molecules with catechol-derivatized hyaluronic acid (HA). The permeable shell of hydrophilic HA chains effectively protects the enzyme from degradation in the blood after intravenous administration and provides an additional function for targeting hepatocytes expressing HA receptor (CD44). The structure and catalytic activity of the enzyme molecules in the nanoparticle were not significantly compromised in the nanoparticle. In addition, ultra-small calcium phosphate nanoparticles (USCaP, 2–5 nm) were crystalized and decorated on the surface of the nanoparticle for the efficient endosomal escape after cellular uptake. The SOD-containing nanoparticle fortified with USCaP was used for the treatment of acetaminophen (APAP)-induced fulminant hepatotoxicity and liver injury. The nanoparticle achieved the efficient hepatic cellular delivery of SOD via a systemic administration and resulted in efficient removal of reactive oxygen species (ROS) in the liver and remarkable improvement of APAP-induced hepatotoxicity and liver injury in animals.</P> <P><B>Statement of Significance</B></P> <P>Despite the enormous therapeutic potential, the intracellular delivery of proteins has been limited due to their poor membrane permeability and stability. In this study, we demonstrated an active enzyme-containing nanoparticle functionalized by hyaluronic acid and ultra-small size calcium phosphate nanoparticles (2–5 nm) for targeted cellular delivery of superoxide dismutase (SOD). The nanoparticle was designed to integrate all the essential functions, including serum stability, target specificity, and endosomal escape capability, for a systemic delivery of a therapeutic protein to the cells of the liver tissue. The intravenous administration of the nanoparticle efficiently removes reactive oxygen species (ROS) in the liver and remarkably improves the drug-induced hepatotoxicity and the progress of fulminant liver injury in an acetaminophen-overdose animal model.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Functional Analyses of Two Acetyl Coenzyme A Synthetases in the Ascomycete Gibberella zeae

Lee, Seunghoon,Son, Hokyoung,Lee, Jungkwan,Min, Kyunghun,Choi, Gyung Ja,Kim, Jin-Cheol,Lee, Yin-Won American Society for Microbiology 2011 EUKARYOTIC CELL Vol.10 No.8

<B>ABSTRACT</B><P> Acetyl coenzyme A (acetyl-CoA) is a crucial metabolite for energy metabolism and biosynthetic pathways and is produced in various cellular compartments with spatial and temporal precision. Our previous study on ATP citrate lyase (ACL) in Gibberella zeae revealed that ACL-dependent acetyl-CoA production is important for histone acetylation, especially in sexual development, but is not involved in lipid synthesis. In this study, we deleted additional acetyl-CoA synthetic genes, the acetyl-CoA synthetases ( <I>ACS</I> genes <I>ACS1</I> and <I>ACS2</I> ), to identify alternative acetyl-CoA production mechanisms for ACL. The <I>ACS1</I> deletion resulted in a defect in sexual development that was mainly due to a reduction in 1-palmitoyl-2-oleoyl-3-linoleoyl-rac-glycerol production, which is required for perithecium development and maturation. Another ACS coding gene, <I>ACS2</I> , has accessorial functions for <I>ACS1</I> and has compensatory functions for <I>ACL</I> as a nuclear acetyl-CoA producer. This study showed that acetate is readily generated during the entire life cycle of G. zeae and has a pivotal role in fungal metabolism. Because ACSs are components of the pyruvate-acetaldehyde-acetate pathway, this fermentation process might have crucial roles in various physiological processes for filamentous fungi. </P>

Variation in 8-ketotrichothecenes and zearalenone production by Fusarium graminearum isolates from corn and barley in Korea

( Yin Won Lee,Jeong Ah Seo ) 한국균학회 1997 97 KOREA CHINA JOINT SYMPOSIUM Vol.- No.-

Self-assembled PEGylated albumin nanoparticles (SPAN) as a platform for cancer chemotherapy and imaging

Lee, Jung Eun,Kim, Myung Goo,Jang, Yeon Lim,Lee, Min Sang,Kim, Nak Won,Yin, Yue,Lee, Jong Han,Lim, Su Yeon,Park, Ji Won,Kim, Jaeyun,Lee, Doo Sung,Kim, Sun Hwa,Jeong, Ji Hoon TaylorFrancis 2018 DRUG DELIVERY Vol.25 No.1

<P><B>Abstract</B></P><P>Paclitaxel (PTX) is used as a major antitumor agent for the treatment of recurrent and metastatic breast cancer. For the clinical application of PTX, it needs to be dissolved in an oil/detergent-based solvent due to its poor solubility in an aqueous medium. However, the formulation often causes undesirable complications including hypersensitivity reactions and limited tumor distribution, resulting in a lower dose-dependent antitumor effect. Herein, we introduce a facile and oil-free method to prepare albumin-based PTX nanoparticles for efficient systemic cancer therapy using a conjugate of human serum albumin (HSA) and poly(ethyleneglycol) (PEG). PTX were efficiently incorporated in the self-assembled HSA-PEG nanoparticles (HSA-PEG/PTX) using a simple film casting and re-hydration procedure without additional processes such as application of high pressure/shear or chemical crosslinking. The spherical HSA-PEG nanoparticle with a hydrodynamic diameter of ca. 280 nm mediates efficient cellular delivery, leading to comparable or even higher cytotoxicity in various breast cancer cells than that of the commercially available Abraxane<SUP>®</SUP>. When systemically administered in a mouse xenograft model for human breast cancer, the HSA-PEG-based nanoparticle formulation exhibited an extended systemic circulation for more than 96 h and enhanced intratumoral accumulation, resulting in a remarkable anticancer effect and prolonged survival of the animals.</P>

Erratum: “Chiral magnetoresistance in Pt/Co/Pt zigzag wires” [Appl. Phys. Lett. <b>110</b>, 122401 (2017)]

Yin, Yuxiang,Han, Dong-Soo,Kim, June-Seo,Lavrijsen, Reinoud,Lee, Kyung-Jin,Lee, Seo-Won,Kim, Kyoung-Whan,Lee, Hyun-Woo,Swagten, Henk J. M.,Koopmans, Bert American Institute of Physics 2018 Applied Physics Letters Vol.112 No.24

Chiral magnetoresistance in Pt/Co/Pt zigzag wires

Yin, Yuxiang,Han, Dong-Soo,Kim, June-Seo,Lavrijsen, Reinoud,Lee, Kyung-Jin,Lee, Seo-Won,Kim, Kyoung-Whan,Lee, Hyun-Woo,Swagten, Henk J. M.,Koopmans, Bert American Institute of Physics 2017 APPLIED PHYSICS LETTERS Vol.110 No.12

Development of a Conditional Gene Expression System Using a Zearalenone-Inducible Promoter for the Ascomycete Fungus Gibberella zeae

Lee, Jungkwan,Son, Hokyoung,Lee, Seunghoon,Park, Ae Ran,Lee, Yin-Won American Society for Microbiology 2010 Applied and environmental microbiology Vol.76 No.10

<B>ABSTRACT</B><P>The ascomycete fungus <I>Gibberella zeae</I> is an important plant pathogen that causes fusarium head blight on small grains. Molecular studies of this fungus have been performed extensively to uncover the biological mechanisms related to pathogenicity, toxin production, and sexual reproduction. Molecular methods, such as targeted gene deletion, gene overexpression, and gene fusion to green fluorescent protein (GFP), are relatively easy to perform with this fungus; however, conditional expression systems have not been developed. The purpose of this study was to identify a promoter that could be induced by zearalenone (ZEA) for the development of a conditional expression system in <I>G. zeae</I>. Through microarray analysis, we isolated one zearalenone response gene (<I>ZEAR</I>) whose expression was increased more than 50 times after ZEA treatment. Northern blot analysis showed that the <I>ZEAR</I> transcript dramatically increased after 1 h of ZEA treatment. To determine the utility of the <I>ZEAR</I> promoter, called Pzear, in a conditional expression system, we transformed a <I>Pzear</I>::<I>GFP</I> fusion construct into <I>G. zeae</I>. Our data showed a ZEA concentration-dependent increase in GFP expression. We also replaced the promoter of <I>G. zeae metE</I> (<I>GzmetE)</I>, an essential gene for methionine biosynthesis, with the Pzear promoter. The growth of the <I>Pzear-GzmetE</I> mutant on minimal medium was dependent on the ZEA concentration supplemented in the medium and showed that GzMetE expression was induced by ZEA. This study is the first report of an inducible promoter in <I>G. zeae.</I> Our system will be useful for the characterization of essential gene functions in this fungus through differential and ZEA-dependent gene expression. In addition, the Pzear promoter may be applicable as a biosensor for the detection of ZEA contamination in agricultural products.</P>