• Title/Summary/Keyword: inhibiting activity

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Development of Human Antibody Inhibiting RNase H Activity of Polymerase of Hepatitis B Virus Using Phage Display Technique (Phage Display 기법을 이용한 B형 간염 바이러스 Polymerase의 RNase H 활성을 억제하는 인간 단세포군 항체의 개발)

  • Lee, Seong-Rak;Song, Eun-Kyoung;Jeong, Young-Joo;Lee Young-Yi;Kim, Ik-Jung;Choi, In-Hak;Park, Sae-Gwang
    • IMMUNE NETWORK
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    • v.4 no.1
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    • pp.16-22
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    • 2004
  • Background: To develop a novel treatment strategy for hepatitis B virus infection, a major cause of liver chirosis and cancer, we aimed to make human monoclonal antibodies inhibiting RNase H activity of P protein playing in important role in HBV replication. In this regard, phage display technology was employed and demonstrated as an efficient cloning method for human monoclonal antibody. So this study analysed the usability of human monoclonal antibody as protein based gene therapy. Methods: RNase H of HBV was expressed as fusion protein with maltose binding protein and purified with amylose resin column. Single chain Fv (scFv) phage antibody library was constructed by PCR cloning using total RNAs of PBMC from 50 healthy volunteers. Binders to RNase H were selected with BIAcore 2000 from the constructed library, and purified as soluble antibody fragment. The affinity and sequences of selected antibody fragments were analyzed with BIAcore and ABI automatic sequencer, respectively. And finally RNase H activity inhibiting assay was carried out. Results: Recombinant RNase H expressed in E. coli exhibited an proper enzyme activity. Naive library of $4.46{\times}10^9cfu$ was screened by BIAcore 2000. Two clones, RN41 and RN56, showed affinity of $4.5{\times}10^{-7}M$ and $1.9{\times}10^{-7}M$, respectively. But RNase H inhibiting activity of RN41 was higher than that of RN56. Conclusion: We cloned human monoclonal antibodies inhibiting RNase H activity of P protein of HBV. These antibodies can be expected to be a good candidate for protein-based antiviral therapy by preventing a replication of HBV if they can be expressed intracellularly in HBV-infected hepatocytes.

Protective Effects of Phosphate and ATP Pretreatment on Pb-Inhibiting Photosystem II Activity (연(Pb)에 의한 광계 II 활성억제에 미치는 인산 및 ATP 전처리의 보호효과)

  • 성민웅
    • Journal of Plant Biology
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    • v.25 no.1
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    • pp.21-36
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    • 1982
  • The activity of photosystem II in isolated chloroplast form the leaves of Sedum sarmentosum was measured. The photoreduction rate of DCPIP by photosystem II showed the circadian rhythm with a peak at near midday sample for a continuing fine day and at near afternoon between nidday and sunset sample for a continuing cloudy day in summer. The optimum light intensity of photoreduction by photosystem II in the chloroplast preparation was about 5~9$\times$$10^4$ lux. The saturated light intensity was over 9$\times$$10^1$lux. Photosystem II activity was inhibited by even the lowest concentration of lead. When Pi and ATP of the same concentration as Pb were added to the reaction mixture containing Tris buffer lacking of Pi prior to Pb incubation, photosystem II activity was protected from Pb-inhibiting effect by the pretreament of Pi and ATP. It was assumed that Pb inhibiiton was probably due to one, P-depriving by the precipitates of $Pb_3$ $($Pb_4$)_2$ in the reaction mixture and the other, partially Pb-combing with Pi groups of the active site of photosystem II.

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Free Radical Scavenging Activity and Inhibition of Linoleic Acid Peroxidation of Commercial Tocotrienol Fraction

  • Kim, Joo-Shin;Chung, Hau-Yin
    • Preventive Nutrition and Food Science
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    • v.12 no.3
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    • pp.177-180
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    • 2007
  • Tocotrienols (T3) are minor plant constituents found abundantly in rice bran, which provide a significant source of vitamin E in animal feeds. T3 was reported to have an intrinsic hypocholesterolemic effect by inhibiting HMG-Co A reductase. It has similar antioxidative properties as tocopherols in food and biological system due to their similar chemical structures. However, the antioxidant activity and mechanism of T3 to scavenge free radicals and to inhibit the peroxidation of linoleic acid are less understood. The purpose of this study was to investigate the scavenging effect of T3 on free radicals and its inhibition of peroxide formation. Free radical scavenging activity was monitored by the DPPH (1,1-diphenyl-2-picrylhydrazyl) method whereas inhibition of linoleic acid peroxidation was evaluated using the thiocyanate method. Thiobarbituric acid (TBA) test was used to determine malonaldehyde formation from linoleic acid peroxidation. Free radical scavenging activity increased with increasing concentration levels of T3. T3 exhibited 38.2, 78.6, 92.7 and 96.2% radical scavenging activity at concentrations of 2, 8, 32 and 128 ppm, respectively. At 128 ppm, it was highly effective in inhibiting linoleic acid peroxidation. The activity of T3 evaluated by the thiocyanate method showed low absorbance values indicating a high level of antioxidant activity. All treatments showed similar trends in antioxidant activity when evaluated by both the thiocyanate method and TBA test.

Bacterial Growth-inhibiting Activity of Amniotic Fluid Against E. coli (양수의 대장균에 대한 세균증식 억제효과)

  • Kim, Soo-Yong;Choi, Myung-Sik;Chang, Woo-Hyun;Cha, Chang-Yong
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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    • pp.233-240
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    • 1987
  • The amniotic fluid provides a medium in which the fetus can readily move, cushions him against possible injury and helps him maintain an even temperature. Besides above mentioned functions, investigators reported that human amniotic fluid contains host-resistance factors which prevent bacteria from producing infectious disease and this activity shows difference among human racial groups or bacterial genera, species and strains. 40 amniotic fluid specimens from Korean women in their second and third trimesters of pregnancy were examined for inhibiting the growth of Escherichia coli. And various factors which might affect bacterial growth inhibiting activity such as pH, initial inoculum size, concentration of amniotic fluid, and heat resistance, were also tested using a strongly inhibitory amniotic fluid specimen. Finally plate diffusion tests were carried out using other strongly inhibitory amniotic fluid. The following results were obtained: 1. Of the 40 fluid samples examined, 18 specimens(45%) had inhibitory activity and samples from women in their second trimester of pregnanancy showed non-inhibitory activity(2 specimens). 2. The pH of the fluids varied between 7.43 and 8.33. There was no correlation between pH and inhibitory activity. 3. No. 19 amniotic fluid showed bacteriostatic activity after 24 hours incubation when an inoculum of $10^2$ organisms per milliliter was used, but non-inhibitory with an inoculum of $10^3$ and $10^4$ bacteria per milliliter. 4. The content of amniotic fluid in culture media influenced E. coli growth. At 90 percent, E. coli was inhibited growth but at 10 percent and 50 percent. 5. Inhibitory activity of No. 19 amniotic fluid was retained after heating to $50^{\circ}C$ for 30 minutes or 100^{\circ}C$ for 30 minutes. 6. Plate diffusion tests with No. 27 amniotic fluid showed that 0.7ml amniotic fluid gave clear zone of growth inhibition around the central well but 0.2ml and 0.1ml amniotic fluids were not.

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Anti-oxidation and Anti-aging Effect of Mixed Extract from Korean Medicinal Herbs (한약재 복합 추출물이 항산화 및 항노화에 미치는 영향)

  • Hwang, Jae-Gyu;Yun, Jong-Kuk;Han, Kil-Hwan;Do, Eun-Ju;Lee, Jin-Sang;Lee, Eun-Ju;Kim, Jong-Boo;Kim, Mi-Ryeo
    • The Korea Journal of Herbology
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    • v.26 no.1
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    • pp.111-117
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    • 2011
  • Objectives : The present study was designed to investigate effects of mixed extract from korean medicinal herbs (MIX) on oxidation/reduction reaction-related and aging-related enzyme in vitro. Methods : We performed MTT assay, collagenase inhibition assay, elastase inhibition assay, tyrosinase inhibition assay, DPPH free radical scavenging assay, SOD-like activity and xanthine oxidase inhibition assay. Results : Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. The MIX showed 97% inhibition of collagenase activity, and 64% inhibition of elastase activity at 1 mg/ml concentration of MIX, next only to positive control, which indicate good efficacy for anti-wrinkle ingredient. Also it's treatment showed 34% inhibition of tyrosinase activity, to relate whitening effect, at the same dose of MIX. Antioxidant activities were determined by DPPH radical scavenging, xanthine oxidase (XO) inhibiting activity and SOD-like activity. Also these scavenging, XO-inhibiting and SOD-like activities were measured in 91%, 80%, and 63% inhibition, respectively, at a treated dose of 1 mg/ml, compare to control. Conclusions : These results suggest that possibility of mixed korean medicinal herbs as a functional ingredient for anti-wrinkle and whitening, anti-oxidation and anti-aging cosmetic formula.

Inhibition Effect of the Extracts of Trichloma matsutake Mycelia on Tyrosinase Activity (송이[Tricholoma matsutake]에서 분리된 균사 배양액의 Tyrosinase 억제효과)

  • 우현정;양덕조
    • KSBB Journal
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    • v.18 no.1
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    • pp.45-50
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    • 2003
  • In this study, the optimum conditions for mycelium culture of the mushroom Tricholoma matsutake and the inhibitory effect of the mycelium extracts no tyrosinase activity have been examined. When the extracts of the Tricholoma matsutake mycelia were tested for inhibitory activity on tyrosinase, it was found that the components extracted with ethyl acetate and water showed the highest inhibitory activity. The effect of antioxidants on the growth of mycelium and tyrosinase-inhibiting activity was also investigated. The results showed that tocopherol inhibited the growth in a concentration-dependent manner. In terms of tyrosinase-inhibiting activity, however, tocopherol was found to enhance the inhibitory activity.

Effect of Proline on First Polar Body Formation in Porcine Primary Oocyte

  • Oh, H. J.;Lee, E. J.;B. C. Yang;W. K. Chang;Kim, J. S.;J. K. Lim;Y. K. Yeo;M. A. Della-Fera;Park, Y. S.
    • Korean Journal of Animal Reproduction
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    • v.26 no.2
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    • pp.165-171
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    • 2002
  • Follicular fluid (FF) contains an oocyte maturation inhibitor with unknown chemical properties. This study was carried out to chemically define the factor(s) inhibiting cumulus cell denudation (CD) and first polar body formation (PBF). Porcine FF (PFF) was extracted with methanol and the extract was serially separated using gel filtration on Superose 12 and Superdex columns. A Superdex fraction was derived with PITC and analyzed with an amino acid analysis column. The results obtained are as follows; PFF had an activity inhibiting both CD and PBF of porcine primary oocytes. Superdex fractions RV2.11 prepared from PFF exhibited an activity inhibiting CD and PBF. By amino acid analysis, the fraction RV2.11 appeared to be proline having the same activity inhibiting CD and PBF. In conclusion, PFF had oocyte maturation inhibitors, of which proline should inhibit CD and PBF.

Tyrosinase Inhibitor from the Flowers of Impatiens balsamina

  • Lim, Young-Hee;Kim, In-Hwan;Seo, Jung-Ju;Kim, Jeong-Keun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1977-1983
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    • 2006
  • Kaempferol was isolated and identified from the methanol extract of the flowers of Impatiens balsamina. Kaempferol showed inhibitory activity against mushroom tyrosinase with an $ID_{50}$ of 0.042 mM. Inhibition kinetics, as determined using a Lineweaver-Burk plot, showed kaempferol to be a competitive inhibitor of mushroom tyrosinase with a $K_i$ value of 0.011 mM. The lag phase of tyrosine hydroxylation catalyzed by mushroom tyrosinase clearly increased on increasing the concentration of kaempferol. In addition to its tyrosinase inhibiting activity, kaempferol strongly inhibited melanin production by Streptomyces bikiniensis, in a dose-dependent manner, without inhibiting cell growth. For comparative purposes, the tyrosinase inhibitory activity of kaempferol was also assayed versus quercetin, a positive standard.

In Vitro α-Amylase, α-Glucosidase, Pancreatic Lipase, Xanthine Oxidase Inhibiting Activity of Agaricus bisporus Extracts

  • Jung Han Kim;Myoung Jun Jang;Youn Jin Park
    • Mycobiology
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    • v.51 no.1
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    • pp.60-66
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    • 2023
  • In this study, the α-amylase inhibitory activity, α-glucosidase inhibitory activity, pancreatic lipase inhibitory activity, and Xanthine Oxidase inhibitory activity of the fruiting body extracts of 5 varieties of Agaricus bisporus (AB) were confirmed. First, the α-amylase inhibitory activity of AB12, AB13, AB18, AB34, and AB40 methanol extracts was lower than that of acarbose, a positive control, in all concentration ranges. The α-glucosidase inhibitory activity of the AB40, AB13, and AB12 methanol extracts at the extract concentration of 1.0 mg/mL was 80.5%, 81.3%, and 78.5%, respectively, similar to that of acarbose, a positive control. The pancreatic lipase inhibitory activity of the methanol extract of Agaricus bisporus fruiting body was significantly lower than that of the positive control orlistat in the concentration range of 50~1.000 (mg/mL). The Xanthine Oxidase inhibitory activity was 0.5~8.0 mg/mL of each extract, which was significantly lower than that of the positive control allopurinol in the same concentration range. However, the Xanthine Oxidase inhibitory activity of AB13 and AB40 at 8.0 mg/mL was about 70%, which was higher than that of other mushrooms. In conclusion, five kinds of Agaricus bisporus fruiting bodies seem to have inhibitory effects on enzymes such as α-amylase, α-glucosidase, pancreatic lipase, and Xanthine Oxidase that degrade starch and protein. In particular, it has an inhibitory effect and a reduction effect on xanthine oxidase that causes gout, so it is expected that it can be developed and used as a food or health supplement with health functional properties through future research.