Biochemical analysis of recombinant CYP4A11 allelic variant enzymes: W126R, K276T and S353G

Han, S.,Cha, G.S.,Chun, Y.J.,Lee, C.H.,Kim, D.,Yun, C.H. JAPANESE SOCIETY FOR THE STUDY OF XENOBIOTICS 2016 DRUG METABOLISM AND PHARMACOKINETICS Vol.31 No.6

Human CYP4A11 is the major ω-hydroxylase of fatty acids in the liver and kidneys. It produces 20-hydroxyeicosatetraenoic acid as well as hydroxylates fatty acids. In this study, we investigated the biochemical properties of three alleles of CYP4A11: W126R, K276T, and S353G. Site-directed mutagenesis of the wild type CYP4A11 was performed, to construct the W126R, K276T, and S353G variant clones. The CYP4A11 wild type and variant constructs were heterologously expressed in Escherichia coli. CO-binding spectra showed the expression of the wild type, K276T and S353G variants, indicating the functional P450 holoenzyme. The W126R variant was not expressed in E. coli. Binding affinities of lauric acid in K276T and S353G variants were stronger than that of wild type. Steady-state kinetics in the hydroxylation reaction of fatty acids were studied. The catalytic efficiencies (k<SUB>cat</SUB>/K<SUB>m</SUB>) of K276T and S353G variants in the reactions without cytochrome b<SUB>5</SUB> were approximately 2- and 4-fold higher, respectively, than that of wild type, and in the reactions with cytochrome b<SUB>5</SUB> they were approximately 2- and 3-fold higher, respectively. These results suggest that individuals carrying the alleles, K276T and S353G, might exhibit higher catalysis of CYP4A11, which may affect the endogenous metabolic products associated with regulation of blood pressure.

PLUG/PLAY 방식 고속 지능형 가공 시스템의 연구

윤원수,김찬봉,권용찬,한기상,양희구,김세광,김주한,박종권 한국공작기계학회 2000 한국공작기계학회 추계학술대회논문집 Vol.2000 No.-

This study aims at developing the high speed/intelligent machining system using the plug/play method of an open architecture controller. The plug/play technology by the Application Specific Function (ASF), can readily implement the open architecture controller into various machining system or other automatic devices. The plug/play method integrates the ASF, visual builder, controller OS technology. This study, as an example, presents a schematic diagram for integration of an open architecture CNC and individual component technology for the high speed/intelligent machining system.

표면항원 및 유전자분석에 의한 V. vulnificus균의 계통분류학적 연구(Ⅰ) : 지방산 조성 및 RAPD PCR에 의한 V. vulnificus균의 계통분류학적 연구

윤영희,김기상,신영학,이점규,오경수,유정식,이상원,이근용 국립보건연구원 1998 국립보건원보 Vol.35 No.-

Pinoresinol-4,4'-di-O-β-d-glucoside from Valeriana officinalis root stimulates calcium mobilization and chemotactic migration of mouse embryo fibroblasts

Do, K.H.,Choi, Y.W.,Kim, E.K.,Yun, S.J.,Kim, M.S.,Lee, S.Y.,Ha, J.M.,Kim, J.H.,Kim, C.D.,Son, B.G.,Kang, J.S.,Khan, I.A.,Bae, S.S. G. Fischer 2009 Phytomedicine Vol.16 No.6

Lignans are major constituents of plant extracts and have important pharmacological effects on mammalian cells. Here we showed that pinoresinol-4,4'-di-O-β-d-glucoside (PDG) from Valeriana officinalis induced calcium mobilization and cell migration through the activation of lysophosphatidic acid (LPA) receptor subtypes. Stimulation of mouse embryo fibroblast (MEF) cells with 10μM PDG resulted in strong stimulation of MEF cell migration and the EC<SUB>50</SUB> was about 2μM. Pretreatment with pertussis toxin (PTX), an inhibitor of G<SUB>i</SUB> protein, completely blocked PDG-induced cell migration demonstrating that PDG evokes MEF cell migration through the activation of the G<SUB>i</SUB>-coupled receptor. Furthermore, pretreatment of MEF cells with Ki16425 (10μM), which is a selective antagonist for LPA<SUB>1</SUB> and LPA<SUB>3</SUB> receptors, completely blocked PDG-induced cell migration. Likewise, PDG strongly induced calcium mobilization, which was also blocked by Ki16425 in a dose-dependent manner. Prior occupation of the LPA receptor with LPA itself completely blocked PDG-induced calcium mobilization. Finally, PDG-induced MEF cell migration was attenuated by pretreatment with a phosphatidylinositol 3-kinase (PI3K) inhibitor such as LY294002. Cells lacking downstream mediator of PI3K such as Akt1 and Akt2 (DKO cells) showed loss of PDG-induced migration. Re-expression of Akt1 (but not Akt2) completely restored PDG-induced DKO cell migration. Given these results, we conclude that PDG is a strong inducer of cell migration. We suggest that the pharmacological action of PDG may occur through the activation of an LPA receptor whereby activation of PI3K/Akt signaling pathway mediates PDG-induced MEF cell migration.

Effects of electron beam irradiation on six insect pests in different sections of flower boxes for export

Yun, S.H.,Koo, H.N.,Kim, H.K.,Cho, S.,Kim, G.H. 한국응용곤충학회 2015 Journal of Asia-Pacific Entomology Vol.20 No.3

We explored effects of electron beam irradiation on disinfestation of six floriculture insect pests (Liriomyza trifolii, Spodoptera litura, Myzus persicae, Tetranychus urticae, Bemisia tabaci, and Frankliniella intonsa) placed at top, middle, or bottom section in boxes of roses and chrysanthemums. After irradiation with an electron beam of 200Gy, eggs of T. urticae, B. tabaci, and F. intonsa were prevented from hatching at every position in the boxes, whereas some eggs of L. trifolii and S. litura hatched at the bottom of the boxes. The pupation and emergence of L. trifolii and S. litura larvae and B. tabaci nymphs were inhibited at every position in the boxes. However, the emergence of T. urticae and M. persicae nymphs was not inhibited, even at the top of the boxes. When pupae were irradiated, the emergence of L. trifolii was inhibited at every section in the boxes, whereas S. litura was not inhibited completely, even at the top section. When adult T. urticae were irradiated, the hatching of the F<SUB>1</SUB> generation was completely inhibited at the middle section in rose boxes but not completely in chrysanthemum boxes. The insect pests that were not inhibited completely at 200Gy were completely inhibited at 300Gy, except for T. urticae. Therefore, the doses of electron beam irradiation required might depend on the types of flowers, the species of insect pests, and the insect pest sections within the boxes.

Lipoteichoic acid of Streptococcus mutans interacts with Toll-like receptor 2 through the lipid moiety for induction of inflammatory mediators in murine macrophages

Hong, S.W.,Baik, J.E.,Kang, S.S.,Yun, C.H.,Seo, D.G.,Han, S.H. Pergamon Press 2014 Molecular immunology Vol.57 No.2

Streptococcus mutans is a pathogenic Gram-positive bacterium that is closely associated with dental caries and subsequent pulpal inflammation. Although lipoteichoic acid (LTA) is considered a major virulence factor of Gram-positive bacteria, little is known about the innate immunity to S. mutans LTA. In this study, we purified LTA from S. mutans (Sm.LTA) through n-butanol extraction, hydrophobic interaction column chromatography, and ion-exchange column chromatography to investigate its immunological properties using murine macrophages. The Sm.LTA preparation had no detectable contamination with endotoxins, proteins, or nucleic acids. Upon exposure to Sm.LTA, the murine macrophage cell-line RAW 264.7 cells produced TNF-α and nitric oxide (NO) in a dose-dependent manner. Sm.LTA preferentially bound to and activated CHO/CD14/TLR2 cells rather than CHO/CD14/TLR4 cells, which are stable transfectants expressing CD14 and TLR2 or CD14 and TLR4, respectively. Sm.LTA could not induce TNF-α or NO production in macrophages derived from TLR2-deficient mice whereas it dose-dependently induced those inflammatory mediators in wild-type macrophages. TLR2-dependent induction of NO by Sm.LTA was also confirmed in RAW 264.7 cells using specific antibodies blocking TLR2. Furthermore, Sm.LTA deacylated by alkaline hydrolysis neither stimulated TLR2 nor induced TNF-α or NO production, suggesting that Sm.LTA lipid moieties are crucial for the immuno-stimulatory activity of Sm.LTA. Unlike Staphylococcus aureus LTA, which has potent immuno-stimulating activity, Sm.LTA showed a modest induction of NO production comparable to LTAs of other oral bacteria Enterococcus faecalis and Lactobacillus plantarum. In conclusion, our results suggest that the Sm.LTA interacts with TLR2 through the lipid moiety for the induction of inflammatory mediators in macrophages.

Identification of stathmin 1 expression induced by Epstein–Barr virus in human B lymphocytes

Baik, S. Y.,Yun, H. S.,Lee, H. J.,Lee, M. H.,Jung, S. E.,Kim, J. W.,Jeon, J. P.,Shin, Y. K.,Rhee, H. S.,Kimm, K. C.,Han, B. G. Blackwell Publishing Ltd 2007 Cell proliferation Vol.40 No.2

<P>Abstract. </P><P><I>Introduction</I>: The Epstein–Barr virus transforms resting B cells into proliferating lymphoblastoid cells, the origin of cell lines. <I>Method and results</I>: Our cDNA microarray analyses led to the identification of 232 up-regulated and 112 down-regulated genes with more than a 3-fold difference in lymphoblastoid cell lines compared to resting B cells. The functional classification of these genes exhibited the distinct expression signature for cell proliferation, cell cycle and an immune response. Among them, we verified the differential expression of several oncogenes such as <I>stathmin 1</I> (<I>STMN1</I>), <I>RAB27A</I>, <I>RAB9A</I>, <I>BACH1</I> and <I>BACH2</I> using quantitative real-time reverse transcriptase-polymerase chain reactions or Western blot analysis. Expression of <I>STMN1</I> (which is involved in regulation of the microtubule filament system, cell growth and S-phase of cell cycle) was increased in lymphoblastoid cell line as well as in 7-day post-Epstein–Barr virus infection B cells, compared to resting B cells. <I>Conclusion</I>: Thus, this study suggests that Epstein–Barr virus infection induces <I>STMN1</I> expression, which play a role in cell cycle progression and proliferation in the human B lymphocyte.</P>

Cartilage Oligometric Matrix Protein-Angiopoietin-1 Promotes Revascularization Through Increased Survivin Expression in Dermal Endothelial Cells of Skin Grafts in Mice

Byun, S.J.,Choi, K.S.,Park, S.H.,Cho, N.W.,Hyun Yoo, C.,Yun, K.J.,Koh, Y.J.,Koh, G.Y.,So, B.J.,Yoon, K.H. American Association of Pathologists and Bacteriol 2007 The American journal of pathology Vol.171 No.5

The present study examined the effects of cartilage oligometric matrix protein angiopoietin-1 (COMP-Ang1) on the revascularization of mice skin grafts. Full-thickness skin grafts were autotransferred into BALB/c mice. The donor grafts were soaked in COMP-Ang1 protein (50 μg/ml, n = 10) or in bovine serum albumin (BSA) (50 μg/ml, n = 10) dissolved in 1 ml of sterile, phosphate-buffered saline for 5 minutes before transfer. Revascularization of the grafts was monitored using an intravital microscope on postoperative days 3, 4, and 5. Morphological and immunohistochemical analyses were performed to evaluate platelet-endothelial cell adhesion molecule-1 and survivin expression and apoptotic signal in the transplanted grafts. Grafts soaked in COMP-Ang1 (COMP-Ang1 group) showed significantly increased revascularization compared with grafts soaked in BSA (BSA group) on intravital microscopy and platelet-endothelial cell adhesion molecule-1 staining. The COMP-Ang1 group showed a significant increase of survivin expression in the endothelial cells and a reduction of apoptotic signal in comparison to the BSA group. Therefore, we believe that COMP-Ang1 provides the therapeutic benefit of enhancing the survival of vascular endothelial cells during transplantation of skin graft.

햄·소시지제품에 대한 소비자 의식 및 구매 실태

조수현,박범영,진구복,유영모,채현석,안종남,이종문,윤상기 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.2

본 설문조사는 국내 육가공제품의 시장에 대한 소비자 의식과 구매형태를 파악하고 시장확대를 위한 기본방안 구축을 위하여 10월부터 12개월까지 3개월 조사한 결과이다. 육가공제품의 구입횟수는 월 1∼2회로 구입하는 소비자가 40.73%로 가장 많았고 그 다음이 주 1∼2회 구입하는 것(28.07%)으로 나타났다. 햄과 소시지를 구분할 수 있는지에 대한 질문에 구분한다가 86.75%로 나타난 반면 구분하지 못한다가 13.25%로 나타났다. 햄과 소시지를 구분하지 못하는 소비자들은 그 이유로서 두 제품의 고기 함량 및 첨가내용물들이 비슷하여서라는 응답이 47.19%로 가장 많았다. 햄과 소시지를 구입할 때 가장 중요하게 고려하는 사항은 회사명(brand name)인 것으로 나타났으며(32.76%) 원료육의 종류(16.53%), 유통기한(16.38%) 및 원료육 생산지(16.23%)인 것으로 나타났다. 햄 또는 소시지 구입시 제품설명 표시내용을 읽느냐는 질문에 응답자들 중 72.25%가 그렇다고 응답한 반면 27.25%는 읽지 않는다고 응답하였다. 소비자들은 햄과 소시지 구입시 불만사항으로 소비자들이 제품내에 고기보다는 결착제를 비롯한 다른 첨가제들이 많아서 고기맛을 거의 느낄 수 없다고 응답하였으며(47.65%), 외관 및 내용물이 비슷한 제품이 너무 많다고 응답한 소비자 및 맛이 없다고 응답한 소비자가 각각 27.70%, 11.62%인 것으로 나타났다. 햄 및 소시지 제품에 대한 요구사항으로 고기 함량이 높은 제품을 생산하기를 원한다고 응답한 소비자가 64.04%였다. 햄과 소시지 제품 판매시 품질에 따른 가격차등제 실시에 대하여 찬성한다고 응답한 소비자가 83.54%인 것으로 나타나 현재 많은 소비자들이 햄과 소시지 제품의 품질에 따른 가격차등제 실시를 원하는 것으로 나타났다. 우리나라도 품질에 따른 등급 기준 설정과 품질별 가격차등제 실시를 통하여 육가공 제품시장의 확대가 가능할 것으로 판단되며 이는 국내돈육의 비선호부위의 소비촉진도 가능할 것이다. Consumer survey(n=1,343) was carried out to investigate the consumer's perception, purchase behavior and demand on the processed meat products. Most consumers purchased the ham and sausage products once a month (40.73%). Consumers(47.19%) didn't distinguish between ham and sausage products due to the similarity of meat contents and additives in products. Consumers(72.75%) considered the product label when they purchased whereas 27.25% of consumers were not interested in the product label. Consumers(47.65%) were not satisfied with ham and sausage products because there were too much additives other than meat, there were many similar processed products in kinds and appearance(27.70%) and they could hardly detect meat taste(11.62%). Consumers demanded to produce the ham and sausage products which contain high meat contents(64.04%). Consumers were favored with establishment of the price differential system depending on the quality based on meat contents in the product(83.54%). In conclusion, the establishment of the price differential system depending on quality as well as the quality grading system for processed meat products is needed not only to increase the processed meat market but also increase the pork consumption especially for non-preferred pork portions.

Identification and functional characterization of novel CYP2J2 variants: G312R variant causes loss of enzyme catalytic activity

Lee, S. S.,Jeong, H.-E.,Liu, K.-H.,Ryu, J.-Y.,Moonb, T.,Yoon, C. N.,Oh, S. J.,Yun, C.-H.,Shin, J.-G. LIPPINCOTT WILLIAMS WILKINS 2005 Pharmacogenetics and genomics Vol.15 No.2