• Title/Summary/Keyword: GLC

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Taurine Concentrations Are Closely Associated with Fatty Acids Concentrations in Breast Milk from Koreans (한국인 모유의 지방산 조성과 타우린 농도의 상관관계에 관한 연구)

  • 박태선
    • Journal of Nutrition and Health
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    • v.31 no.1
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    • pp.88-95
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    • 1998
  • Kong chain $\omega$3 & $\omega$6 fatty acids and taurine have been suggested to have structural and /or functional roles in the brain. The purpose of this study was to evaluate the changes in the percentage of fatty acids and taurine concentration in human milk obtained from Korean mothers(n=37) as lactation proceeds from birth to 5wks postpartum , and to determine the correlation between taurine concentration and fatty acid composition in breast milk. Level of fatty acids and taurine in breast milk were measured by GLC and HPLC, respectively. The percentages of medium -chain saturated fatty acids(10 : 0, 12 : 0 & 14: 0) were increased significantly , while percentages of long-chain saturated fatty acids(22 :0 & 24 : 0), and most of the long chain $\omega$6 and $\omega$3 fatty acids(20 : 3$\omega$6 , 20 :4$\omega$6, 22 :4$\omega$6 , 22 :5$\omega$6, 22 :5$\omega$3 & 22 : 6$\omega$3) were decreased significantly in mature milk (5 wks postpartum ) as compared to the colostrum. Taurine concentrations in colostrum and mature milk from Korean mothers were 549 $\pm$58 and 233 $\pm$41nmol/ml , respectively. Taurine concentrations in human milk was negatively correlated with the percentages of medium -chain saturated fatty acids ( 10 : 0 &12 : 0), and positively correlated with the percentages of long-chain saturated fatty acids( 16 :0, 20 : 0, 22 : 0& 24 : 0) and long-chain monounsaturated fatty acids( 20 : 1, 22 : 1 & 24 ; 1). Taurine concentration in human milk was positively correlated with elongation indices of both $\omega$6 and $\omega$3 fatty acids, and negatively correlated with the desaturation index of $\omega$6 fatty acids. These close relationships between fatty acids compositions and taurine level in human milk are worth pursing further investigation, especially with regard to their common roles in retina and brain development.

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Screening System for Chitin Synthase II Inhibitors from Natural Resources and its Inhibitor Prodigiosin

  • Hwang, Eui-Il;Kim, Young-Kook;Lee, Hyang-Bok;Kim, Hong-Gi;Kim, Sung-Uk
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.251-257
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    • 2000
  • Chitin synthases are identified as key enzymes of chitin biosynthesis in most of the fungi. Among them, chitin synthase II has been reported to be and essential enzyme in chitin biosynthesis, and exists as a membrane-bound form. To search and screen new antifungal agents from natural resources to inhibit chitin synthase II, the assay conditions were established using the enzyme isolated from Saccharomyces cerevisiae ECY38-38A(pAS6) that overproduces only chitin synthase II. This enzyme was activated only by partial proteolysis with trypsin. Its actibity reached the maximum at $80{\;}\mu\textrm{g}/ml$ of trypsin and was strongly stimulated by 2.0 mM $Co^{2+}$, 1.0 nM UDP-[$^{14}C$]-GicNAc, and 32 mM free-GlcNAc. Under these assay conditions, the highest chitin synthase II activity was observed by incubation at $30^{\circ}C$ for 90 min. However, and extremely narrow range of organic solvents up to as much as 25% of DMSO and 25% of MeOH was useful for determining optimal assay conditions. After a search or potent inhibitors of chitin synthase II from natural resources, prodigiosin was isolated from Serratia marcescens and purified by solvent extration and silica gel column chromatographies. The structure of prodigiosin was determined by UV, IR, Mass spectral, and NMR spectral analyses. Its molecular weight and formula were found to be 323 and $C_{20}H_{25}N_{3}O$, respectively. Prodigiosin ingibited chitin synthase II by 50% at the concentration of $115{\;}\mu\textrm{g}/ml$.

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Novel Modification of Growth Medium Enables Efficient E. coli Expression and Simple Purification of an Endotoxin-Free Recombinant Murine Hsp70 Protein

  • Zachova, Katerinat;Krupka, Michal;Chamrad, Ivo;Belakova, Jana;Horynova, Milada;Weigl, Evzen;Sebela, Marek;Raska, Milan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.7
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    • pp.727-733
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    • 2009
  • Heat shock protein 70 kDa (hsp70), a molecular chaperone involved in folding of nascent proteins, has been studied for its ability to activate innate and specific immunity. High purity hsp70 preparation is generally required for immunization experiments, because endotoxins and other immunologically active contaminants may affect immune responses independently of hsp70. We have developed a novel modification of E. coli-expression medium that enabled a simple two-step production and purification method for endotoxin-free recombinant hsp70. During Ni-NTA-based affinity purification of hsp70, a contaminating protein from host E. coli cells, L-glutamine-n-fructose-6-phosphate aminotransferase (GFAT), was identified. By testing various compounds, supplementation of growth medium with a GFAT metabolite,N-acetylglucosamine, was found to reduce GFAT expression and increase the total hsp70 yield five times. The new protocol is based on column purification of His-tagged hsp70 protein produced by E. coli with the modified medium, followed by endotoxin removal by Triton X-114 extraction. This approach yielded hsp70 with high purity and minimal endotoxin contamination, making the final product acceptable for immunization experiments. In summary, a simple modification of growth medium allowed production of recombinant mouse hsp70 in high yield and purity, thus compatible with immunological studies. This protocol may be useful for production of other Histagged proteins expressed in E. coli.

Analysis of Organochlorine Pesticide Residues in the Presence of Polychlorinated Biphenyls (PCBs) -Part II. Analysis of River Sediments and Cultivating Soils in the Peripheries of Several Industrial Estates- (Polychlorinated Biphenyls(PCBs) 존재하(存在下)에 유기염소계(有機鹽素系) 살충제의 잔류분(殘留分) 분석(分析) -제2보(第2報), 공단주변(工團周邊)의 하천(河川) 및 전답토양(田畓土壤) 분석(分析)-)

  • Park, Chang-Kyu;Park, No-Dong
    • Applied Biological Chemistry
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    • v.23 no.1
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    • pp.58-63
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    • 1980
  • Present investigation is undertaken to evaluate residues of organochlorine pesticides and PCBs in river sediments and cultivated soils by GLC. Main sampling areas chosen were three industrial estates, Suwon, Iri and Kumi. Samples consist of river sediments and cultivated soils, in Suwon, Iri and Chilgok, which are likely to be accumulated with PCBs from nearby factories. Organochlorine pesticide residues were found in all sediments and soil samples while no PCBs residue was detected in all samples analyzed. The pesticide residues were greatest in the Suwon and least in Chilgok soils. p,p'-DDT and BHC isomers $({\alpha}-,\;and\;{\gamma}-)$ were found in all soil samples. p,p'-DDT, at the range of 0.006-0.840 ppm, was the most abundant residues among organochlorine insecticides.

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Changes of Volatiles from Apple Fruits during Maturity and Storage -Part. I. Identification and Determination of Volatiles in the Fruits- (사과 성숙(成熟) 및 저장중(貯藏中) 향기성분(香氣成分)의 변화(變化) -제일보(第一報). 과실(果實) 성숙중(成熟中) 향기성분(香氣成分)의 동정(同定) 및 정량(定量)-)

  • Shim, Ki-Hwan;Sohn, Tae-Hwa;Kim, Myung-Chan;Kang, Shin-Kwoen;Park, Seok-Kyu
    • Applied Biological Chemistry
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    • v.27 no.1
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    • pp.14-20
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    • 1984
  • Volatiles from Mallus pumila Miller Var. Fuji, Ralls Janet and Jonathan during maturity were examined. The volatiles of apples were collected by the headspace trapping method using. Tenax GC and identified by gas liquid chromatography and GC-mass spectrometry. Among over 34 GLC peaks, 23 esters, 9 alcohols and 2 aldehydes were identified. The quantity of volatiles produced was increased in proportion to the degree of maturity, and 1-butanol, isobutyl butyrate, 2-pentanol, ethyl valerate and hexanal were major.

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Autoxidative Stability of Triglyceride Molecular Species (트리글리세리드 분자종의 산화안정성에 관한 연구)

  • Yoon, Hyeung-Sik;Kim, Seon-Bong;Park, Yeung-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.18 no.2
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    • pp.205-210
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    • 1989
  • The influence of triglyceride molecular species on autoxidation was investigated by determining the residual molecular species after incubating soybean oil triglycerides. The molecular species of soybean oil triglycerides were analyzed by capillary column gas chromatography and electron impact ionization mass spectrometry utilizing selected ion monitoring. The autoxidative stability of each molecular species in soybean oil triglycerides appeared to decrease in proportion to the increase in the number of double bonds present in the acyl residues, and it was affected by degree of unsaturation of fatty acid when the number of double bonds in triglyceride were the same. And it appeared to be enhanced by a decreases in the length of the saturated acyl chain present in the glycerides.

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Serum Stress Responses during Seawater Acclimation in Coho Salmon, Oncorhynchus kisutch (은연어의 해수 순치에 따른 스트레스 반응의 변화)

  • Hong, Kyung-Pyo;Kim, Pyong-Kih;Jeon, Joong-Kyun;Kim, You-Hee;Park, Yong-Ju;Myoung, Jung-Goo;Kim, Jong-Man
    • Ocean and Polar Research
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    • v.26 no.3
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    • pp.433-438
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    • 2004
  • Stress response to the seawater acclimation in coho salmon (Oncorhynchus kisutch) smolt was investigated. Salt concentration of rearing water was gradually increased for 3 days from freshwater to seawater (30 ppt salt level). The changes of serum concentrations of cortisol as a primary stress indicator, and as secondary indicators, glucose (GLC), lactate (LAC), triglyce.ide (TG), cholesterol (CHOL), sodium ion $(Na^+)$, chloride ion $(Cl^-)$ and enzyme activities (alanine aminotransferase, ALT: aspartate aminotrasferase, AST; lactate dehydrogenase, LDH) were quantified during the acclimation experiment. Among them, cortisol, LAC, TG, CHOL, ALT, AST concentrations showed rapid increase at the first exposure to the 10ppt salt level (day 1), and began to decrease to the constant values after day 2 of adaptation at 20ppt salt level. However, LDH concentration tended to decrease during the whole experimental period. $Na^+\;and\;Cl^-$ showed slight decrease at day 1, and increased to a little bit higher values after day 2 rather than those in freshwater. All the fishes started on taking a food after day 4 of seawater adaptation. From these results, to reduce osmotic shock inducible stress to fish in seawater acclimation, gradual increase of salt levels is recommended.

A Study on the Production Medium of Lactic Acid Bacterial Cells by Using Corn Steep Liquor (Corm Steep Liquor를 이용한 젖산균이 생산배지에 관한 연구)

  • 안영태;김근배;인영민;정석근;함준상;김동운;이경욱;김선기;김현욱
    • Food Science of Animal Resources
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    • v.20 no.3
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    • pp.181-191
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    • 2000
  • 젖산균의 생장을 위한 질소, 탄소 공급원으로서 corn steep liquor의 이용 가능성을 시험하고 반응 표면 분석(Response surface metho-dology)을 이용하여 젖산균의 최적 생상 배지 조성을 연구하였다. 반응 표면 분석에서 L.fermentum의 생장배지에 첨가되 corn steep liquor와 yeast extract의 농도(p<0.01) 그리고 corn steep liquor와 yeast extract의 교호작용(P<0.05)이 L.ferm-entum의 생장에 큰 영향을 미치는 것으로 나타났으며, 이때 생균수가 최대인 corn steep liquor의 함량은 10.77%, yeast extract는 3.39%.Tween 80은 1.69%으로 예측되었다. 한편, Lc, lactis ssp. lactis 의 생장배지는 corn steep liquor 의 농도(P<0.01) 그리고 corn steep li-quor와 $\beta$-glycerophosphate disodium salt의 교호작용(P<0.05)인 Lc. lactis ssp. lactis의 생장에 큰 영향을 미치는 것으로 나타났으며 이때 생균수가 최대인 corn steep liquor의 함량은 3.5%, $\beta$-glycerophosphate disodium salt는 4.38%로 예측되었다. MRS broth와 예측된 최적 배지에서 L.fermentum의 젖산과 초산의 생성량은 각각 0.166, 0.114과 0.273, 0.081 M이고 M 17glc broth와 최적배지에서 Lc. lacti ssp. lactis의 젖산과 초산의 생성량은 각각 0.089, 0.003과 0.189, 0.003M이었다. 따라서 corn steep liquor는 L. fermentum와 Lc. lactis ssp, lactis 의 생장을 위해 질소 또는 탄소 공급원으로서 배지에 첨가 될 수 있는 우수한 농업 부산물로 판단되었다.

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Generation of Female Porcine Fibroblasts Expressing Efficiently Membrane Cofactor Protein at ${\alpha}1$,3-Galactosyltransferase locus (${\alpha}1$,3-Galactosyltransferase 유전자 좌위에서 Membrane Cofactor Protein을 효과적으로 발현하는 자성 돼지 섬유아세포의 생산)

  • Oh, Keon Bong;Kim, Bella;Hwang, Seongsoo;Ock, Sun-A;Im, Seoki;Park, Jin-Ki
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.289-295
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    • 2013
  • Xenotransplantation of pig organs into primates results in fatal damage, referred as hyperacute rejection (HAR), and acute humoral xenograft rejection (AHXR), to the organ graft mediated by antibodies pre-existing and newly-producing in primates against their cognate pig antigens. Functional ablation of ${\alpha}1$,3-galactosyltransferase (Gal-T KO) of pig which is an enzyme involved in synthesis of Gala1-3Galb1-4GlcNAc-R antigen is essentially required to prevent HAR. Moreover, additional genetic modification under Gal-T KO background for enforced expression of human complement regulatory proteins which can inhibits complement activation is known to effectively imped HAR and AHXR. In this study, we constructed a membrane cofactor protein (MCP) expression cassette under control of human $EF1{\alpha}$ promoter. This cassette was inserted between homologous recombination regions corresponding to Gal-T locus. Subsequently this vector was introduced into ear skin fibroblasts of female pig by nucleofection. We were able to obtained 40 clones by neomycin selection and 4 clones among them were identified as clones targeted into Gal-T locus of MCP expression cassette by long-range PCR. Real time RT-PCR was shown to down-regulation of Gal-T expression. From these results, we demonstrated human $EF1{\alpha}$ promoter could induce efficient expression of MCP on cell surface of fibroblasts of female pig.

Hexose Uptake and Kinetic Properties of the Endogenous Sugar Transporter(s) in Spodoptera frugiperda Clone 21-AE Cells

  • Lee Chong-Kee
    • Biomedical Science Letters
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    • v.11 no.3
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    • pp.327-332
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    • 2005
  • Sf21 cells become popular as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains $0.1\%$ D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike human glucose transporters, very little is known about the characteristics of the endogenoussugar transporter(s) in Sf21 cells. Thus, some kinetic properties of the sugar transport system were investigated, involving the uptake of 2-deoxy-D-glucose (2dG1c). In order to obtain a true measure of the initial rate of uptake, the uptake of $[^3H]2dGlc$ from both low $(100{\mu}M)$ and high (10 mM) extracellular concentrations was measured over periods ranging from 30 sec to30 min. The data obtained indicated that the uptake was linear for at least 2 min at both concentrations, suggesting that measurements made over a 1min time course would reflect initial rates of the jexpse uptake. To determine $K_m\;and\;V_{max}$ of the endogenous glucose transporter(s) in Sf21 cells, the uptake of 2dG1c was measured over a range of substrate concentrations $(50{\mu}M\~10mM)$ 2dG1c uptake by the Sf21 cells appeared to involve both saturable and non-saturable (or very low affinity) components. A saturable transport system for 2dG1c was relatively high, the $K_m$ value for uptake being < 0.45 mM. The $V_{max}$ value obtained for 2dG1c transport in the Sf21 cells was about 9.7-folds higher than that reported for Chinese hamster ovary cells, which contain a GLUT1 homologue. Thus, it appeared that the transport activity of the Sf21 cells was very high. In addition, the Sf21 glucose transporter was found to have very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter

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