• Bulletin of the Korean Chemical Society
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• v.17 no.11
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• pp.1018-1022
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• 1996

Adenosine deaminase (ADA) has been utilized as the label in devising a potentiometric homogeneous assay for riboflavin and riboflavin binding protein (RBP). The proposed homogeneous assay method employs an ADA-biotin conjugate as the signal generator and an avidin-riboflavin conjugate as the signal modulator in the solution phase. The catalytic activity of the ADA-biotin conjugate is inhibited in the presence of an excess amount of the avidin-riboflavin conjugate, and the observed inhibition is reversed in an amount proportional to the concentration of RBP added. When the analyte riboflavin is added to this mixture of ADA-biotin, avidin-riboflavin and RBP, the activity of the enzyme conjugate is re-inhibited in an amount proportional to the concentration of riboflavin. Since the enzyme label used in this system is ADA, an ammonia-producing enzyme, a potentiometric rather than photometric detection scheme is used to monitor the enzymatic activity in the assay.

• Korean journal of food and cookery science
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• v.27 no.3
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• pp.1-10
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• 2011

The antioxidant activity and cytotoxic effect of an ethanol extract from Seoritae were analyzed to develop new functional food materials. The antioxidant activity of Seoritae was determined by measuring electron donating ability with 1,1-diphenyl-2picrylhydrazyl (DPPH) and 2-2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assays, as well as the ferric reducing antioxidant power (FRAP) assay. The cytotoxic effect of the Seoritae ethanol extract was measured with the 3-(4,5-dimethylthiazol-2-yl)-2,5-dipheltetrazolium (MTT) and sulforhodamine B (SRB) assays. As a result, the electron donating abilities of Seoritae against the DPPH and ABTS radicals were 63.75% and 87.68% at 500 ${\mu}g$/assay, respectively. The $IC_{50}$ values of Seoritae in the DPPH and ABTS assays were 385.39 ${\mu}g$/assay (128.46 ${\mu}g/mL$) and 209.39 ${\mu}g$/assay (51.83 ${\mu}g/mL$). Additionally, the FRAP value of Seoritae was 0.84 $FeSO_4$ eq. mM at 800 ${\mu}g$/assay. The total amounts of polyphenols and flavonoids, which indicate the antioxidant capability of Seoritae extract were 1.65 mg/g and 0.59 mg/g, respectively. Moreover, Seoritae extract showed a high cytotoxic effect of up to 81% against human cancer cells, particularly A-549 and HeLa cells. The growth inhibition rate of Seoritae extract against A-549 and HeLa cells was up to 76.48% and 75.67% in the MTT assay, and 78.98% and 80.54% in the SRB assay, respectively. The results of this study suggest that an ethanol extract of Seoritae is a potentially good natural antioxidant.

• Journal of Acupuncture Research
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• v.24 no.5
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• pp.23-32
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• 2007

Objectives : Angiogenesis consists of the proliferation, migration, and differentiation of endothelial cells, and angiogenic factors and matrix protein interactions modulate this process. The aim of this study was to determine whether herbs medicine(KHBJs) could induce angiogenic activity in human umbilical vein endothelial cells(HUVECs). Methods : The angiogenic activity of KHBJs were evaluated by proliferation using BrdU assay, chemotactic migration assay, tube formation assay, and measurement of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor(VEGF) in HUVECs. Also, In order to identify enhance angiogenic activity by activity guided fractionation, the angiogenic activity of fractions of KHBJs such as KHBJB or KHBJR were evaluated in vitro and in vivo Matrigel plug angiogenesis asaay. Results : About 9 KHBJs significantly increased HUVECs proliferation in a dose-dependent manner. In addition, 9 herbs medicine(KHBJs) increased migration and tube-like formation in HUVECs. Interestingly the expression of bFGF and VEGF, an angiogenesis-inducing growth factor, were dose-dependently increased by KHBJs. However, angiogenic activity of fractionated KHBJs(KHBJB or KHBJR) not enhanced more than KHBJs in HUVECs and Matrigel plug in vivo angiogenesis assay. Conclusions : 9 KHBJs significantly induces angiogenesis in in vitro and in vivo. These results suggest that 9 KHBJs potent angiogenic agents and promising drug for the induction of neovascularization.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.27 no.1
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• pp.17-27
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• 2001

A cell-based assay system for monitoring NF-$textsc{k}$B activity was developed to determine the influence of activated NF-$textsc{k}$B in human HaCaT cells. The pNF-$textsc{k}$B-SEAP-NPT plasmid that permits expression of the secreted alkaline phosphatase (SEAP) reported gene in response to the NF-$textsc{k}$B activity and contains neomycin phosphotransferase (NPT) gene for the geneticin resistance in host cells was constructed and transfected into human keratinocyte cell line HaCaT. Human HaCaT transfectant cells secreted the SEAP enzyme into the culture medium in a time-dependent manner until 72h. NF-$textsc{k}$B activities were measured in the SEAP reporter gene assay using a fluorescent detection method. The treatment of HaCaT cell transfectants with known antioxidants [e.g., N-acetyl-L-cysteine and vitamin C] showed inhibition of NF-$textsc{k}$B activity in a time-and concentration-dependent manner. The phorbol 12-myristate 13-acetate (PMA) known as a stimulator of NF-$textsc{k}$B expression demonstrated that it increased NF-$textsc{k}$B activity in a time- and concentration-dependent manner. This assay system could be used to determine the quantitative measurement of NF-$textsc{k}$B activity in the human skin and allow the screening of anti-inflammatory agents from various synthetic chemicals and natural products for dermatological purpose. Abbrevitions used: NF-$textsc{k}$B, nuclear factor kappa B; I-$textsc{k}$B, Inhibitory kappa B; SEAP, secreted alkaline phosphatase; NPT, neomycin phosphotransferease; PCR, polymerase chain reaction: dNTP, deoxynucleoside triphosphates; DMEM, dulbecco’s modified eagle medium; FBS, fetal bovine serum; PBs, phosphate-buffered saline; MUP, 4-methylumbellifery phosphate; NAC, N-acetyl-L-cysteine; DMSO, dimethyl sulfoxide; PMA, phorbol 12-myristate 13-acetate.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.255-255
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• 2002

For rapid and sensitive measurement of antiviral activities, application of a recombinant virus containing firefly luciferase gene was attempted. Recombinant human cytomegalovirus (HCMV) containing luciferase gene driven by HCMV late gene pp28 promoter (HCMV/pp28-luc) was used to test the antiviral activities of three known compounds and the result was compared with results from the conventional plaque assay for measuring the production of infectious viruses. When human fibroblast cells were infected with HCMV/pp28-luc, luciferase activity was observed at 2 days after infection and reached maximum at 6 days after infection, whereas the production of infectious virus was maximal at 4 days after infection. The antiviral activities of ganciclovir, acyclovir, and papaverine were measured in HFF cells infected with HCMV/PP28-luc and the luciferase activity was compared with the infectious virus titers. Luciferase activity decreased as the concentration of ganciclovir or papaverine increased, while there was a slight decrease in luciferase activity with acyclovir. The level of the decrease in Luciferase activity was comparable to the level of decrease in the production of infectious virus. Therefore, the antiviral assay using recombinant virus HCMV/pp28-luc resulted in sensitivity similar to the conventional plaque assay with a significant reduction in assay time.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.1
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• pp.137-147
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• 2004

In order to evaluate the antitumor activity and immunomodulatory effects of Seoleosojong-tang(SST), studies were done. We measured the cytotoxic activity for various kinds of cancer cells, inhibitory effect on activity of DNA topoisomerase I, cell adhesion to complex extracellular matrix, survival time in ICR bearing S-180, pulmonary colonization and histological changes of lung in C57BL/6 injected i.v. with B16-F10, CAM assay, expression of CD4/sup +/, CD8/sup +/, B220/sup +/, cytokine gene in spleen cell. The results were obtained as follows: 1. In cytotoxicity against A549, HT1080, 816-F10, NCL-H661 was showed cytotoxicity as compared with control. 2. The inhibitory effect on adhesion of A549, 816-F10 to complex extracellular matrix was over 40% at 100 ㎍/㎖ of SST. 3. In DNA topoisomerase I assay, SST has inhibitory effect. 4. The T/C% was 120.8 in SST treated group in S-180 bearing ICR mice. 5. In pulmonary colonization assay, a number of colonies were decreased significantly and histological changes were showed that infiltration area of cancer cells were inhibited effectively in SST treated group. 6. In CAM Assay, SST has antiangiogenic effect. 7. On the expression of positive cell to CD4/sup +/, CD8/sup +/ and 8220/sup +/ in spleen cells, CD4/sup +/ cells were increased significantly in SST treated group. 8. Effect of SST on IL-1β gene expression in splenic cell was significantly increased as function of whole concentration. 9. The gene expression of IL-4, IL-6, IL-10, IL-12, IFN-γ, TNF-α were increased in SST treated group. From above results SST could be usefully applied for antitumor activity and immunomodulatory effects, but further research of SST should be required.

• Archives of Pharmacal Research
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• v.29 no.11
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• pp.1049-1054
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• 2006

Activation of Sphingosine kinase (Sphk) increases a bioactive lipid, sphingosine 1-phosphate (S1P) and has been observed in a variety of cancer cells. Therefore, inhibition of Sphk activity was an important target for the development of anticancer drugs. As a searching tool for Sphk inhibitor, we developed fluorescent Sphk activity assay combined with high performance liquid chromatography (HPLC). Previously we established murine teraticarcinoma mutant F9-12 cells which lack S1P lyase and stably express Sphk1. By using F9-12 cells, optimal assay conditions were established as follows; $100\;{\mu}M\;of\;C_{17}-Sph\;and\;30\;{\mu}g$ protein of F9-12 cells lysate in 20 min. Sphingosine analog $C_{17}-Sph$ was efficiently phosphorylated by Sphk activity ($K_{m}:67.08\;{\mu}M,\;V_{max}\;:1507.5\;pmol/min/mg$). New product $C_{17}-S1P$ was separated from S1P in reversed-phase HPLC. In optimized conditions, 300 nM of phorbol 12-myristate 13-acetate (PMA) increased Sphk activity approximately twice while $20\;{\mu}M$ of N,N-dimethylsphingosine (DMS) reduced 70% of Sphk activity in F9-12 cells lysate. In conclusion, we established non-radioactive but convenient Sphk assay system by using HPLC and F9-12 cells.

• The Korean Journal of Food And Nutrition
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• v.24 no.4
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• pp.496-500
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• 2011

The purpose of this investigation was to evaluate anti-tumor activity and detect what compounds affect its activity form $Alpinia$ $officinarum$ Hance. Two fractions, methanol and ethylacetate, were isolated by Amberlite XAD-2 resin column chromatography from methanol extract of the rhizomes of $Alpinia$ $officinarum$. In hollow fiber assay, the methanol extract and methanol fraction were found to inhibit the tumor growth against colon tumor cell lines such as Colo-320, HCT116 and WiDr. Three diarylheptanoids [5-hydroxy-1,7-diphenyl-3-heptanone, 5-hydroxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone and 3,5-dihydroxy-1,7-diphenylheptane] and two flavonoids [galangin and kaempheride] were isolated and identified from the methanol fraction, which is higher activity than ethylacetate fraction. Among these diarylheptanoids and flavonoids, 3,5-dihydroxy-1,7-diphenylheptane, galangin and kaempheride as active components on anti-tumor activity were mainly posited in methanol fraction.

• Mycobiology
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• v.40 no.2
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• pp.134-137
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• 2012

Mushrooms collected from Deogyu mountain, Korea, in 2011, were identified as four classes, four orders, 13 families, 22 genera, and 33 species. In particular, agaricales was most abundant and comprised more than 70%. Their antioxidant activities were estimated using three different bioassay methods, the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) radical scavenging assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, and reducing power assay. As a result, the methanol extracts of Stereum ostrea, Laetiporus sulphureus var. miniatus, and Tyromyces sambuceus exhibited potent antioxidant activity in all bioassays tested.

• Korean Journal of Pharmacognosy
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• v.44 no.3
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• pp.224-229
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• 2013

Bromotopsentin (BSM) is a bisindole alkaloid compound, which is recognized as a metabolite of the marine sponge Spongosorites sp. In this study, the antioxidant activity of BSM was investigated. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, the trolox equivalent antioxidant capacity (TEAC) assay, the superoxide radical scavenging (NBT) assay, the lipid peroxidation and hydroxyl radical-induced DNA damage assays were carried out to evaluate the antioxidant activity of BSM. It was found that BSM had stronger scavenging activity on the stable free radical DPPH and superoxide radical than L-ascorbic acid with an $IC_{50}$ value of 62 and 64 ${\mu}M$, respectively. The TEAC value which indicated the total antioxidant capacity of BSM was about 0.8, which was also stronger than L-ascorbic acid. About 1.3 ${\mu}M$ of BSM induced 50% inhibition of lipid peroxidation. 60 nM of BSM exhibited a significant protective activity against DNA strand scission by hydroxyl radical on pBR322 DNA. Taken together, we suggest that BSM possesses strong antioxidant activity, and could be a valuable new addition to the list of anti-aging chemotherapeutic agents.