• Title/Summary/Keyword: Binding medium

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Study on The Preventive Effect of Ginsenosides Against Hypercholesterolemia and Its Mechasnism (인삼사포닌의 고 cholesterol 혈증 강하작용에 관한 연구)

  • 윤수희;주충노
    • Journal of Ginseng Research
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    • v.17 no.1
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    • pp.1-12
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    • 1993
  • The Preventive effect of the saponin fraction of Panax ginseng C.A. Meyer against hyperchole- sterolemia was demonstrated by assaying the cholesterol and triacylglyceride level of the blood serum and liver of rats fed high-cholesterol diet with and/or without ginsenoside. To understand the mechanism of the preventive action of ginsenoside, ginsenoside effect on LDL receptor binding ability, cholesterol level, and cAMP level of Chinese hamster ovary (CHO) cells cultured under various conditions were examined. When LDL (20 $\mu$g/ml) was added to the culture medium for CHO cell culture, LDL receptor binding activity of the cell was suppressed down to 58% of that of normal group. Ginsenosides at 10--2% and 10-3% level (w/v) were shown to exert an appreciable stimulatory effect on CHO cell LDL receptor activity, which partially overcame the suppression due to the presence of LDL (20 $\mu\textrm{g}$/ml) in the medium. Ginsenosides also reduced the increased cholesterol level of test group almost to that of normal group, and it increased cAMP level, which was usually reduced to 55% of that of the normal group due to the presence of LDL in the medium. Comparison of Kd and Bmax value of CHO cells cultured under different conditions revealed that this stimulation was due not to the receptor's binding affinity but to its number. Addition of ginsenoside (10-2%) decreased the uptake of taurocholic acid as much as 20% at the actively transporting everted ileal sacs, but it failed to form a large mixed micelles with taurocholic acid, which was one of the proposed mechanisms by which ginsenoside inhibits bile acid reabsorption. From the above results, it seemed likely that ginsenoside prevented hypercholestrolemia by decreasing cholesterol level in cells thereby relieving the inhibition of LDL receptor synthesis by cholesterol and also by inhibiting bile acid reabsorption from the small intestine.

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Aluminium and Cadmium Interfere with the Estrogen Receptor Level in the Primary Culture of Hepatocytes in the Rainbow Trout Oncorhynchus mykiss

  • Hwang Un-Gi
    • Fisheries and Aquatic Sciences
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    • v.4 no.4
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    • pp.180-185
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    • 2001
  • Al and Cd-induced inhibition of vitellogenin (VTG) production was examined at the estrogen receptor (ER) level in rainbow trout Oncorhynchus mykiss hepatocytes. The binding of $[^3H]$ $estradiol-17\beta\;(E_2)$ to hepatocytes reached a plateau 3 days after addition of $E_2\;(2\times\;10^{-6} M)$to the medium. The binding activity was linearly reduced with the increased concentrations $(-10^{-5}\;M)$ of 4-hydroxy-tamoxifen (4-OHT) and specific binding linearly increased with the increased doses of $[^3H]\;E_2$, indicating that the radioligand bound to ER. Al $(-10^{-4}\;M)$and Cd $(10^{-6}\;M)$ as well as 4-OHT $(10^{-6}\;M)$ significantly reduced the $[^3H]\;E_2$-binding activity by $30­40\%$, while they completely inhibited VTG production. Al and Cd had no effect on $E_2-human$ $ER\alpha$ binding activity at any concentrations used $(-10^5\;nM\;each)$. These results suggested that Al and Cd inhibited VTG production in part by interfereing with the ER level. Inhibitory effects of these metals on the $E_z-dependent$ upregulation of ER activity are also discussed.

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Esterified-Glucomannan in Broiler Chicken Diets-Contaminated with Aflatoxin, Ochratoxin and T-2 Toxin: Evaluation of its Binding Ability (in vitro) and Efficacy as Immunomodulator

  • Raju, M.V.L.N.;Devegowda, G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1051-1056
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    • 2002
  • In vitro binding efficacy of esterified glucomannan (E-GM) (0.1%) on aflatoxin B1 (AF) (300 ppb), ochratoxin A (OA) (2 ppm) and T-2 toxin (T-2) (3 ppm), when present alone or in combination, was evaluated in toxin-contaminated feed at pH 4.5 and 6.5. Esterified glucomannan showed significantly (p<0.01) higher binding with AF (81.6%), whereas those recorded with T-2 (27.8%) and OA (25.6%) were moderate. Binding of each toxin decreased as the number of toxins in feed increased. pH of medium showed no effect on mycotoxin binding ability of E-GM. A $2{\times}2{\times}2{\times}2$ factorial experiment of 5 week duration was conducted to study the effects of two dietary levels each of AF (0 and 300 ppb), OA (0 and 2 ppm), T-2 (0 and 3 ppm ) and E-GM (0 and 0.1%) on the immune competence of a total of 960 day-old commercial broilers. Reductions in size of thymus (by AF and T-2) and bursa (by AF) and antibody titers against Newcastle disease and Infectious Bursal disease (by all the toxins) were noted. Additive and antagonistic interactions were seen among the toxins on certain parameters. Esterified glucomannan significantly (p<0.01) improved antibody titers and weights of bursa ofFabricius and thymus indicating its counteracting efficacy against immunosuppression in mycotoxicosis of multiple origin.

Change of hCG Binding Capacity on the Granulosa Cell of Porcine Ovary During Follicular Atresia (돼지 난소내 여포의 폐쇄과정중 과립세포의 결합능의 변화)

  • Chang, Choul-Soo;Lee, Chang-Joo;Yoon, Yong-DaI;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.13 no.1
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    • pp.29-38
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    • 1986
  • In order to study the mechanism of follicular atresia, follicles were classified into the normal groups and the atretic ones, according to the criteria with or without corpus luteum, size of follicles, vascularization, status of granulose cells and the hypertrophy of theca layers in the porcine ovary. To estimate the binding capacity of human chorionic gonadotropin (hCG) receptor on the granulosa cells during atresia, hCG were iodinated by the chloramine-T method and then purified through the column chromatography. The concentration" of hCG receptor in each group were measured by the hCG receptor binding assay. Binding capacity in large normal follicles were 1.16%, but atretic ones were 0.45%. But in medium and small follicles (below 6mm in diameter), the binding capacity in normal follicles were 0.09%, but atretic ones were 0.05%, which was lower than those of large follicles. The present ( ) that the concentrations of hCG receptors on granulosa cells is decreased when the follicles become atretic and be used as a sort of creteria for the identification of follicles atresia.

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Intragenic Suppressors for Expory-defective Signal Sequence Mutation of Ribose-binding Protein in Escherichia coli (대장균 리보스 결합단백질의 신호배열 변이에 대한 숙성체 부위의 회복돌연변이)

  • 이영희;송택선;김정호;박순희;박찬규
    • Korean Journal of Microbiology
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    • v.29 no.5
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    • pp.270-277
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    • 1991
  • A mutational alteration in the signal sequence of ribose-binding protein (RBP) of Escherichia coli, rbsB103, completely blocks the export of the protein to the periplasm. Intragenic suppressors for this mutation have been selected on minimal medium with ribose as a sole carbon source. Six suppressor mutations were characterized in detail and were found to have single amino acid wubstitution in the mature portion of RBP, which resulted in the mobility shift of the proteins on SDS polyacrylamide gel. Amino acid changes of these suppressors were localized in several peptides which are packed to form the N terminal domain of typical bilobate conformation of RBP. The involvement of SecB, a molecular chaperone, was investigated in the suppression of signal sequence mutation. Translocation efficency was found to be increased by the presence of SecB for all suppressors. It is likely that the folding characteristics of RBP altered by the suppressor mutations affect the affinity of interaction between SecB and RBP.

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Effects of $\beta$-Mercaptoethanol on lipid Peroxidation and Fertilization Ability In Vitro by Xanthine-Xanthine Oxidase System in Pig (Xanthine-Xanthine Oxidase System,하에서 돼지 동결-융해정자의 Lipid Peroxidation과 체외수정능력에 대한 $\beta$-Mercaptoethanol의 영향)

  • 사수진;정희태;이장희;유일선;양부근;김정익;박춘근
    • Korean Journal of Animal Reproduction
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    • v.26 no.3
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    • pp.263-273
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    • 2002
  • This study was undertaken to evaluate the effects of $\beta$-mercaptoethanol ($\beta$-ME) on lipid peroxidation and fertilization ability in vitro by xanthine (X) - xanthine oxidase (XO) system in boar spermatozoa frozen-thawed. The boar spermatozoa were treated with X and/or XO, and the spermatozoa viability were measured by the eosin-nigrosin stain method. In control group, level of vitality in boar spermatozoa were higher than in medium with X, XO and X+XO groups. No significant differences, however, were observed under the all conditions. The percentage of spermatozoa that reached acrosome reaction were significantly (P<0.05) higher in sperm treated without that than with $\beta$-ME under the all conditions. On the other hand, when spermatozoa were inseminated in medium with X and/or XO, the penetration rates in all conditions were higher in medium with that than without $\beta$-ME. However, significant differences were not observed between medium with and without $\beta$-ME. The lipid peroxidation of sperm was evaluated on the basis of malondialdehyde (MDA) production. The MDA were higher in sperm treated without that than with $\beta$-ME under the above all conditions. However, significant differences were not observed between medium with and without $\beta$-ME. Sperm-SH group were higher detected in medium with that than without $\beta$-ME under the all conditions. The activity of sperm binding to Bona pellucida was also evaluated through binding to salt-stored porcine oocytes. In control group, sperm binding to zona pellucida were significantly (P<0.05) higher than in medium with X+XO groups. The sperm binding in all conditions were higher in medium with that than without $\beta$-ME. However, significant differences were not observed between medium with and without $\beta$-ME. These results suggest that addition of $\beta$-ME in X-XO system may play a positive role in improving of fertilization ability in vitro.

19F NMR Investigation of F1-ATPase of Escherichia coli Using Fluorinated Ligands

  • Jung, Seun-Ho;Kim, Hyun-Won
    • BMB Reports
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    • v.31 no.1
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    • pp.44-47
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    • 1998
  • Asymmetry amongst nucleotide binding sites of Escherichia coli $F_1$-ATPase was examined using $^{19}F$ NMR signal from fluorinated analogs of adenine nucleotides bound to nucleotide binding sites. ADP-$CF_2-{PO_3}^{2-}$ showed no inhibitory effect to $F_1$-ATPase. But ADP-CHF-${PO_3}^{2-}$ (racemic mixture) showed competitive inhibition of $F_1$-ATPase with $K_i$ of $60\;{\mu}m$. ADP-CHF-${PO_3}^{2-}$ shows only negligible binding to $EF_1$ in the absence of $Mg^2+$. With the addition of $Mg^2+$ to the medium, the $^{19}F$ resonance of free ADP-CHF-${PO_3}^{2-}$ disappeared and the new broad resonances appeared. Appearance of more than two new asymmetric resonances following the binding of ADP-CHF-${PO_3}^{2-}$ to $EF_1$ may indicate that at least one of the isomers showed split resonances. This may suggest that the region between ${\alpha}$-and ${\beta}$-phosphate of ADP-CHF-${PO_3}^{2-}$ which is bound to catalytic sites is experiencing a different environment at different sites.

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Studies on pathogenicity of Escherichia coli isolated from chickens: Correlation between in vitro Congo-red binding properties and in vivo virulence in avian Escherichia coli (닭유래 Escherichia coli의 병원성에 관한 연구 : 시험관내 Congo-red 결합능과 병원성간의 상관관계)

  • Woo, Yong Ku;Kim, Ki Seuk;Kim, Bong Hwan
    • Korean Journal of Veterinary Research
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    • v.31 no.1
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    • pp.55-61
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    • 1991
  • The correlation between in vitro Congo-red binding properties of E coli and in vivo invasiveness of the organisms in SPF chickens and mice was investigated. Congo-red positive E coli colonies were dark-red color with a typical colonial morphology of rough appearance when grown on Congo-red medium, while Congo-red negative colonies showed pale-pink color and smooth surfaced colonial morphology. Pathogenicity of 10 Congo-red positive E coli for mice was observed in 92.5% but that of 5 Congo-red negative E coli was 45%. Invasiveness of 10 Congo-red positive E coli for chickens was observed in 96% of the SPF chickens tested but that of 5 Congo-red negative E coli was 16% only. These results of pathogenicity studies with E coli isolates indicate a significant correlation between Congo-red binding ability and virulence in avian Escherichia coli.

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Studies on Cd and Removal Ability and Detoxification of Oenanthe stolonifera (미나리 ( Oenanthe stolonifera ) 의 Cd, Zn 제거능과 내성에 관한 연구)

  • Lee Soo;In Sook Lee
    • The Korean Journal of Ecology
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    • v.19 no.6
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    • pp.519-527
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    • 1996
  • To examine the possibility of biomonitoring of heavy metal removal ability and soil, a study was performed to investigate the heavy metal removal ability and metal-binding protein (MBP) as detoxification process using Oenanthe stolonifera. After O. stolonifera was exposed to individuals (cadmium, zinc) and mixture (cadmium+zinc)for 4 days, removal rate of heavy metal and pH in the treatment medium was measured. MBP was assayed by means of ion exchange column chromatography. The exposure to mixture (Cd:76.8%, Zn:75%) rather than individuals (Cd:82.9%, Zn:90.4%) showed a synergism raising the toxic effect. Initial removal rate was different for each heavy metal : in case of exposure to cadmium it was over 60% on day 1, while for zinc it was 75~90% on day 4. Throughout the experimental period, pH value of treatment medium continuously decreased, since cortex in the roots may secret organic acid to adjust and prevent toxicity of metals. The existence or MBP in the 70~80 fraction and the presence of Zn-enzyme pool was ascertained with the column chromatography. This study demonstrated a possibility that heavy utilized as a biomarker of heavy metal pollution.

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Effects of Individual Fatty Acids on Receptor-Mediated Binding, Internalization and Degradation of $[^{125}I]LDL$

  • Choue, Ryo-Won;Cho, Byung-Hee Simon
    • BMB Reports
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    • v.30 no.1
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    • pp.1-6
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    • 1997
  • The ability of Hep-G2 cells to process $[^{125}I]LDL$ under basal conditions was investigated. The receptor-binding and internalization of $[^{125}I]LDL$ increased with the time of incubation in a saturable manner. After 4 h of incubation, 31.4 ng of $[^{125}I]LDL$ was cell bound. The cells rapidly internalized $[^{125}I]LDL$ via specific, receptor-mediated endocytosis. The amount of internalized $[^{125}I]LDL$ reached a maximun of 96.7 ng at 2 h of incubation and remained constant for the next 2 h. The rate of degradation of internalized $[^{125}I]LDL$ proceeded in a linear manner over the entire 4 h of incubation after an initial lag period. The effects of individial fatty acids (C18:0. C18:1, C18:2. and C18:3), differing in their degree of unsaturation. on the receptor-binding, internalization and degradation of $[^{125}I]LDL$ were also investigated. Inclusion of 1.0 mM of each fatty acid into the culture medium significantly increased $[^{125}I]LDL$ metabolism in Hep-G2 cells. Among the fatty acids tested, stearic acid had the least effect on the receptor-binding activity. There were no significant differences among the unsaturated fatty acids in LDL-receptor binding. The effect of individual fatty acids on the $[^{125}I]LDL$ uptake was similar to that of the receptor-binding. showing a significantly lower effect with stearic acid. The amount of degraded material of internalized $[^{125}I]LDL$ was the lowest with stearic acid when it was compared with unsaturated fatty acids.

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