• Title/Summary/Keyword: NR assay

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Taxol Production by an Endophytic Fungus, Fusarium redolens, Isolated from Himalayan Yew

  • Garyali, Sanjog;Kumar, Anil;Reddy, M. Sudhakara
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1372-1380
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    • 2013
  • Different endophytic fungi isolated from Himalayan Yew plants were tested for their ability to produce taxol. The BAPT gene (C-13 phenylpropanoid side chain-CoA acetyl transferase) involved in the taxol biosynthetic pathway was used as a molecular marker to screen taxol-producing endophytic fungi. Taxol extracted from fungal strain TBPJ-B was identified by HPLC and MS analysis. Strain TBPJ-B was identified as Fusarium redolens based on the morphology and internal transcribed spacer region of nrDNA analysis. HPLC quantification of fungal taxol showed that F. redolens was capable of producing $66{\mu}g/l$ of taxol in fermentation broth. The antitumour activity of the fungal taxol was tested by potato disc tumor induction assay using Agrobacterium tumefaciens as the tumor induction agent. The present study results showed that PCR amplification of genes involved in taxol biosynthesis is an efficient and reliable method for prescreening taxol-producing fungi. We are reporting for the first time the production of taxol by F. redolens from Taxus baccata L. subsp. wallichiana (Zucc.) Pilger. This study offers important information and a new source for the production of the important anticancer drug taxol by endophytic fungus fermentation.

Effects of Sophorae Radix Extract in Pulmonary Vascular Endothelial Cells Damaged by XO/HX (고삼 추출물이 XO/HX에 의해 손상된 혈관내피세포에 미치는 영향(I))

  • Kwon Kang Beom;Lee Ho Seung;Kim In Su;Kim In Gyu;Ryu Do Gon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.549-552
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    • 2003
  • To investigate the protective effect of Sophorae Radix (SR) on the damage by pulmonary vascular endothelial cells by xanthine oxidase (XO)/hypoxanthine (HX)-induced oxygen tree radical, Neutral Red (NR) and c-fos immunopositive cell assay were used. The results were obtained as follows ; The viability of vascular endothelial cells treated with XO/HX was decreased. And c-fos immunopositive cells represented a maximal increase in group treated with XO/HX for 2 hour in pulmonary vasvular endothelial cells. But pretreated groups with SR extracts were not inhibited the increase of c-fos immunopositive cells by XO/HX in a dose-dependent manner. These results show that XO/HX elicits toxic effects in cultured pulmonary vascular endothelial cells, and suggest that SR extract is very effective in the prevention of XO/HX-induced increase of c-fos immunopositive cells.

Metal Effects of Urban Air Particulates on Cytokine Production and DNA Damage

  • Lee, Kwan-Hee;Hong, Yun-Chul
    • Toxicological Research
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    • v.17 no.4
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    • pp.255-265
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    • 2001
  • Epidemiologic studies have demonstrated an association between short-term exposure to particulate air pollutants and increased mortality. However the biological mechanism underlying these associations have not been fully established and also the chemical and physical characteristics of the pollutant particles are not well understood. The metal constituents of air pollutant particles and their bioavailability are considered to Play an important role as possible mediators of Particle-induced airway injury and inflammation. Sprague-Dawley rat alveolar macrophage cells (NR8383) were exposed to airborne and acid-leached particulate matter (PM). Titanium oxide and nickel subsulfide were used as negative and positive controls. Particle-induced reactive oxygen species formation in cells was detected using the fluorescent probe 2',7'-dichlorofluorescin diacetate. Expression of TNF-$\alpha$ and IL-6 were measured by enzyme-linked immunosorbent assay, and PM-induced DNA double-strand breaks were determined with $\lambda$DNA/Hind III marker. Metals associated with air pollutant particles mediated intracellular oxidant production in alveolar macrophages, and the cytotoxicity and proinflammatory cytokine production induced by PM were associated with oxidative stress. The oxidants produced by air pollutant particles also are likely to induce DNA double-strand breaks. Our findings in alveolar macrophage cells exposed to PM and acid-leached PM support the hypothesis that metal components in urban air pollutants and their bioavailabilities might play an Important role in the induction of the adverse health effects.

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산삼의 배양 및 그 응용에 관한 연구

  • 신미희
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.27 no.2
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    • pp.45-56
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    • 2001
  • Korea mountain ginseng known as oriental miracle drug is an important medicinal plant. The effect of mountain ginseng adventitious roots extract has been described. The valuable root of mountain ginseng contained several kinds of ginsenosides that have been confirmed to have many active functions for the human body. However, the study of mountain ginseng has a limit because the price of wild ginseng is very expensive and rare. The mountain ginseng adventitious roots were derived from mountain ginseng callus that were induced from mountain ginseng roots. Adventitious roots were separated from callus and grown in solid media(Murachige and stoog media). It was cultured in a 20L bioreactor. After culturing for 40days, adventitious roots were harvested. Afterwards the harvested mountain ginseng adventitious roots were dryed and extracted. We examined the effect on melanogenesis of mountain ginseng adventitious roots extract. Here, we report the inhibitory effect of melanin biosynthesis on the adventitious roots extract of In vitro test. Also, we assessed the safety of adventitious roots extract. In vitro, cytotoxicity of adventitious roots extract was assessed in mouse fibroblast using two method: The neutral red uptake assay and the MTT assay. In vivo, the allergic and irritant were Patch teated in 30 patients. Consequently, extract of mountain ginseng adventitious roots have inhibitory effect on melanin biosynthesis in B-16 melanoma cell test, tyrosinase inhibitory test and DOPA auto-oxidation test. There were decreased 86%(0.5% concentration), 45%(1% concentration) and 61%(1% concentration), respectively.

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Anti-thrombotic Activities of Hot-water Extracts Prepared from Various Parts of Nelumbo nucifera Gaertner (다양한 부위의 연(Lotus) 열수 추출물의 항혈전 활성)

  • Ahn, Seon-Mi;Sung, Hwa-Jung;Kim, Jong-Sik;Park, Jong-Yi;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1156-1162
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    • 2018
  • To investigate the anti-thrombotic activities of the lotus (Nelumbo nucifera Gaertner), various hot-water extracts were prepared from the leaf, pod of seed (PS), seed, embryo of seed (ES), root, and the node of root (NR) of the lotus. The highest extraction ratio was found in the NR (20.3%), followed by the seed, root, leaf, ES, and PS. These extracts had pH and acidity levels ranging from 5.6~6.5 and 0.06~0.20%, respectively. The seed extract showed 70% brix, whereas the leaf and PS extracts showed less than 0.1% brix. The highest contents of total polyphenol (179.7 mg/g), total flavonoids (161.4 mg/g), and reducing sugar (161.4 mg/g) were observed in the leaf extract, and the highest total sugar content (873.0 mg/g) in the seed extract. Anti-coagulation assay of the extracts of NR, leaf, and PS showed strong activities. In particular, at a concentration of 5 mg/ml, the PS extract had 15-fold extended thrombin, prothrombin, and activated partial thromboplastin times. However, only the ES extract showed activities inhibitory to platelet aggregation, with treatment with 0.25 mg/ml of ES extract decreasing platelet aggregation to 25.1%, a reduction comparable to that effected by aspirin. The extracts other than the seed extract showed no hemolysis activities against human RBC at treatments of up to 1 mg/ml. These results suggest that the NR, PS, seed, and ES, all byproducts of the lotus agriculture industry, have high potential as novel sources of anti-thrombotic agent.

Cytotoxicity of Hydrogen Peroxide and Effects of Rhizoma Gastrodiae Against Hydrogen Peroxide in Mouse Cerebral Neurons (생쥐의 배양 대뇌신경세포에 대한 Hydrogen Peroxide의 세포독성 및 천마의 영향)

  • Choi Yu Sun;Lee Eun Mi;Son Young Woo;Lee Kang Chang;Shin Yong Il;Song Myung Su;Choi Young Ja;Choi Kyu Chul;Kang Hyung Won;Lim Chang Yong;Rhu Ti Yong;Park Sea Hong;Park Seung Taeck
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.5
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    • pp.928-931
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    • 2002
  • To elucidate the toxic effect of oxygen free radicals on cultured mouse cerebral neurons damaged by hydrogen peroxide(H₂O₂)-induced neurotoxicity, we examined the neurotoxicity induced by oxygen radicals by NR assay when cultured cerebral neurons were grown in the media containing various concentrations of H202 for 6 hours. In addition, neuroprotective effects of herb extracts such Rhizoma Gastrodiae(RG) on H202-induced neurotoxicity in cultured cerebral neurons were evaluated after cultured cerebral neurons were preincubated with various concentrations of herb extract, RG for 2 hours before 50uM H₂O₂ for 6 hours. H₂O₂ decreased remarkably cell viability in dose-and time-dependent manner in these cultures, and also herb exract, RG decreased LDH activity of cerebral neurons damaged by H₂O₂. From the above results, it is suggested that H₂O₂ was toxic in cultured cerebral neurons from mouse, and RG was effective in blocking the neurotoxicity induced by oxygen radicals in these cultures.

Studies on the Cytotoxicity and Antitumor Activity of Perilla frutescens (소엽의 세포독성 및 항암작용에 관한 연구)

  • Han, Du-Seok;Chung, Boung-Ho;Yoo, Hyeon-Gyeong;Kim, Young-Ok;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.25 no.3
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    • pp.249-257
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    • 1994
  • The cytotoxic and antitumor activity of Perilla frutescens extract on cultured 3T3 fibroblast and skin melanoma cells were evaluated by tetrazolium MTT (MTT) and neutral red (NR) colorimetric assay methods. Lactate dehydrogenase activity was also measured. The light microscopic study was carried out to observe morphological changes of cultured mouse fibroblast and skin melanoma cells. The results were as follows: 1. Water and ether extracts showed a significant cytotoxicity in 3T3 fibroblast and all extracts exhibited a significant anti-tumor activity in skin melanoma cells. Methanol, ethyl acetate and ethanol extracts showed low cytotoxic effects, but exhibited a high anti-tumor activity. 2. The MTT absorbance in 3T3 fibroblast was significantly decreased by treatment with ether, water, chloroform and ethanol extracts and skin melanoma cells was significantly decreased by treatment with all extracts. The difference in MTT absorbance in two cell types was most remarkable when treated with methanol and ethanol extracts. 3. Methanol and ethyl acetate extracts showed the strongest effect in growth inhibition of melanoma cells. These results indicated that methanol extract possessed a low cytotoxicity and a strong anti-tumor activity.

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Study on the Anti-aging Activity of Gentianae sino-ornata Ethanol Extract (용담화 에탄올 추출물의 항노화 작용 연구)

  • Choi, Hyung-Wook;Li, Shun-Hua;Kim, Eun-Joo;Kim, Soo-Kyung;Lee, Jang-Cheon;Lim, Kyu-Sang
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.28 no.2
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    • pp.1-12
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    • 2015
  • Objective : The goal of this study is to identify the effects of extract ofGentianae sino-ornata(GSO) on the anti-oxidative activity of skin.For this purpose, several functions of GSO were analyzed in terms of skin-lightening activity and wrinkle improvement. Methods : Cell viability was measured by neutral red (NR) assay, and GSO showed highly efficacy in DPPH radical scavenging activity. The level of tyrosinase and matrix metalloproteinase-1 (MMP-1) in media was analyzed by ELISA kit, and the expressions of p-JNK and p-ERK was measured by Western blot. To elucidate inhibitory effects of GSO on melanin synthesis, I determined the tyrosinase activity and melanin production in B16F10 cells. Results : MMP-1 production in UVB-stimulated HDF cells was inhibited by GSO treatments, and also GSO inhibited protein expression levels of p-JNK and p-ERK. GSO significantly reduced tyrosinase activity and melanin synthesis in B16F10 cells. Conclusions : From these results, GSO appears to be effective on skin elasticity increase, wrinkle improvement, whitening as anti-aging activity.

Effects of Gamdu-tang Extract in Rat Cardiac Endothelial Cells (심장내피세포의 DNA 합성량에 미치는 감두탕의 영향(I))

  • Kwon Kang Beom;Kim Woo Kyung;Kim In Su;Kang Gil Seong;Kim In Gyu;Kim In Seob;Ryu Do Gon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.352-355
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    • 2003
  • To investigate the protective effect of Gamdu-tang(GDT) and its constituents. Radix Glycyrrhizae(RG) and Semen Glycine(SG) on the damage of cardiac endothelial cells by xanthine oxidase (XO)/hypoxanthine (HX)-induced oxygen free radical, Neutral Red (NR) and DNA synthesis assay were used. The results were obtained as follows ; Cardiac endothelial cells treated with XO/HX showed the cytotoxicity such as decreases in viability and DNA synthesis. Cardiac endothelial cells pretreated with GDT extracts were not showed the decrease of DNA synthesis by XO/HX, These results show that XO/HX elicits toxic effects in cultured cardiac endothelial cells derived from neonatal rat, and suggest that GDT extract is very effective in the prevention of XO/HX-induced toxicity.

Study on Anti-inflammatory Effect of Ethanol Extract of Gentianae sino-ornata (용담화 에탄올 추출물의 항염증 효과에 관한 연구)

  • Choi, Hyung-Wook;In, Myung-Hee;Lee, Shun-Hua;Kim, Eun-Joo;Lim, Kyu-Sang;Woo, Won-Hong
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.28 no.4
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    • pp.51-61
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    • 2015
  • Objective : This research has been conducted to comparative analysis of anti-inflammatory effects to suggest a usefulness of ethanol extracts fromGentianae sino-ornata(GSO) as a functional material for inflammatory activities.Methods : Cell viability was measured by neutral red (NR) assay, and nitric oxide (NO) production in RAW264.7 cells was monitored by measuring the nitrite content in culture medium. The expressions of cyclooxigenase-2 (COX-2) was determined by western blot analysis, and Inducible nitric oxide syntase (iNOS) and cytokine were determined by reverse transcriptase-polymerase chain reaction (RT-PCR).Results : When the GSO extract was added the concentration of 5-20 ㎍/㎕, the viability of cells was maintained 90% or more at all levels. NO production was suppressed by the treatment of GSO in LPS-stimulated RAW264.7 cells. GSO inhibited the expression of iNOS, COX-2, IL-1βand IL-6 in LPS-stimulated RAW264.7 cells.Conclusions: From this results, we consider that GSO can be a useful therapeutic and preventive approach to various inflammatory diseases as a functional material for inflammatory activities.