• Title/Summary/Keyword: NRU assay

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Toxicity Assessment of Gas Phase in Cigarette Smoke Using Cell-free Assay

  • Park, Chul-Hoon;Sahn, Hyung-Ok;Shin, Han-Jae;Lee, Hyeong-Seok;Min, Yaung-Keun;Hyun, Hak-Chul
    • Journal of the Korean Society of Tobacco Science
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    • v.29 no.2
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    • pp.110-117
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    • 2007
  • In vitro toxicity tests such as cytotoxicity, mutagenicity and genotoxicity assay are useful for evaluating the relative toxicity of smoke or smoke condensates obtained from different cigarette configurations. A major disadvantage of these tests is relatively time-consuming, complicated and expensive. Recently, a cell-free glutathione consumption assay (GCA) as a rapid and simple screening method for the toxicity assessment of smoke has been reported by Cahours et al. (CORESTA, 2006). This study was carried out to assess the GCA application capable of predicting the toxicity of gas/vapor phase (GVP) of cigarette smoke and to identify individual compounds responsible for the glutathione (GSH) consumption in smoke. Each GVPs from 2R4F, standard cigarette, carbon filter cigarette (ExC) and new carbon filter cigarette (ExN), test cigarettes were collected by automatic smoking machine and evaluated the relative toxicity by GCA and neutral red uptake (NRU) assay. Toxic compounds existed in smoke were also chosen, relative toxicities of these compounds were screened by using two methods and compared individually. The overall order of toxicity by GCA was 2R4F > ExC > ExN, which was consistent with the result of Neutral Red Uptake assay. The levels of carbonyl compounds of ExN were lower than those of 2R4F and ExC, indicating that GSH consumption was associated with carbonyl compound yields. A major toxicant under current study is acrolein, which contributed to more than half of the GSH consumption. Collectively, the toxicity of GVP determined by GCA method may be mainly attributed to acrolein.

Improvement of Skin Photoaging by Polysaccharide Extract Derived from Tremella fuciformis (White Jelly Mushroom)

  • Choi, Jae-Hwan;Kim, Bora
    • Natural Product Sciences
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    • v.27 no.4
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    • pp.300-306
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    • 2021
  • Chronic ultraviolet (UV) radiation causes photoaging, which represents skin damage, disrupts skin barrier function, and promotes wrinkle formation. We investigated that the polysaccharide extract of an edible basidiomycetous white jelly mushroom, Tremella fuciformis, (TF-Glucan®) exhibited statistically photoprotective activity by inhibiting matrix metalloproteases (MMPs) and increasing collagen synthesis, and an anti-inflammatory activity by inhibiting nitric oxide and pro-inflammatory cytokines at the concentrations of less than 1000 ㎍/ml, which is not cytotoxic (p < 0.05). Additionally, TF-Glucan® increased the expression of involucrin and filaggrin to prevent the disruption of UVB-induced barrier function (p < 0.05). TF-Glucan® was assessed as a safe material by the human primary skin irritation (1, 3, 5%), human repeated insult patch test (no sensitization at 5%), 3T3 NRU phototoxicity assay (no phototoxicity, PIF < 2, MPE < 0.1), eye irritation test test by BCOP (no category, IVIS ≤ 3) and local lymph node assay (negative at 10, 25, 50%) for identifying potential skin sensitizing. These results suggest that TF-Glucan® may be useful as an anti-photoaging ingredient for developing cosmeceuticals.

Preventive Characteristics of Garlic Extracts Using in vitro Model System on Alzheimer's Disease (In vitro model system을 활용한 마늘 추출물의 치매예방 특성)

  • Choi, Gwi-Nam;Kim, Ji-Hye;Kwak, Ji-Hyun;Jeong, Chang-Ho;Jeong, Hee-Rok;Shin, Jung-Hye;Kang, Min-Jung;Sung, Nak-Ju;Heo, Ho-Jin
    • Journal of agriculture & life science
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    • v.44 no.4
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    • pp.45-55
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    • 2010
  • In this study, the acetylcholinesterase (AChE) inhibition and neuronal cell protective effects of water, 100% methanol and dichlromethane extracts from garlic were investigated. We found that dichloromethane extract of garlic resulted in a dose-dependent manner on AChE inhibition ($IC_{50}$: $36.1{\mu}g/mL$). In cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), cell viabilities of water, 100% methanol and dichlromethane extracts were lower (almost under 40%) than amyloid ${\beta}$ protein ($A{\beta}$)-induced neurotoxicity. Because $A{\beta}$ is also known to increase neuronal cell membrane breakdown, neuronal apoptosis was further confirmed by lactate dehydrogenase (LDH) and neutral red uptake (NRU) assay. Water extract presented relative protection against $A{\beta}$-induced membrane damage in LDH assay. However all garlic extracts showed significant problem with decrease of cell viability in NRU assay, especially at dichloromethan extract. To determine active compounds in column fractions (98:2 fraction) from dichloromethane extract which showed significant AChE inhibitory effect, we performed HPLC and LC-MS analysis. It was supposed that garlic may contain allyl methyl disulfide, diallyl monosulfide, and diallyl disulfide as active compounds.

The chemical composition and biological activities of volatile flavor components of Elsholtzia splendens (꽃향유 전초의 향기성분 분석과 생리활성 평가)

  • Jeong, Jae Hoon;Lim, Heung Bin
    • Analytical Science and Technology
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    • v.18 no.6
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    • pp.500-510
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    • 2005
  • This study was conducted to investigate the chemical composition of essential oil, absolute and oleoresin isolated from Elsholtzia splendens and their biological activities. Yields of essential oil, absolute and oleoresin extracted from Elsholtzia splendens were 0.28%, 12.45% and 9.95%, respectively. The major component was 2-cyclohexen-1-one in essential oil, methyl linolenate in absolute and 9,12,15-octadecatrienoic acid in oleoresin. Essential oil and oleoresin showed the inhibitory activities in enzyme-dependent, enzyme-independent and autooxidative lipid peroxidation system. $EC_{50}$ values in nuetral red uptake (NRU) assays for the exposure times of 24 h were $46.4{\mu}g/ml$, $681.7{\mu}g/ml$ and $17.6{\mu}g/ml$ in essential oil, absolute and oleoresin, and oleoresins showed the cytotoxic effect at the only high dose. Any mutagenic and antibiotic activity did not show in absolute and oleoresin, but, there were mutagenic and antibiotic activities only when treated with essential oil $500{\mu}g/ml$ above in Ames test. Essential oil and oleoresin might be somewhat effective in prolongating the ciliostasis of rat trachea.

Antioxidant and Neuronal Cell Protective Effects of Methanol Extract from Schizandra chinensis using an in vitro System (In vitro system에서 오미자 메탄올 추출물의 항산화 및 신경세포 보호효과)

  • Kim, Ji-Hye;Jeong, Chang-Ho;Choi, Gwi-Nam;Kwak, Ji-Hyun;Choi, Sung-Gil;Heo, Ho-Jin
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.712-716
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    • 2009
  • In this study, the antioxidant and neuronal cell protective effects of methanol extract from Schizandra chinensis were evaluated. The proximate composition and total phenolics content of the extract were as follows: 64.88% nitrogen free extract, 10.56% crude fiber, 10.22% moisture, 8.33% crude protein, 5.05% ash, 0.96% crude fat, and 83.04 mg/g of total phenolics. In assays the methanol extract of Schizandra chinensis presented ferric reducing/antioxidant power (FRAP) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity in a dose-dependent manner. In a cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), the methanol extract showed protective effect against $H_2O_2$-induced neurotoxicity, and lactate dehydrogenase (LDH) release into medium was also inhibited by various concentrations of extracts (68-80%). Cell viability after treatment of the methanol extract was higher than that shown for vitamin C ($100\;{\mu}M$) using a neutral red uptake (NRU) assay. Therefore, these data suggest that the methanol extract of Schizandra chinensis may be useful for neurodegenerative diseases including Alzheimer's disease.

Anticancer Activity of Petroselinum sativum Seed Extracts on MCF-7 Human Breast Cancer Cells

  • Farshori, Nida Nayyar;Al-Sheddi, Ebtesam Saad;Al-Oqail, Mai Mohammad;Musarrat, Javed;Al-Khedhairy, Abdulaziz Ali;Siddiqui, Maqsood Ahmed
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5719-5723
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    • 2013
  • Pharmacological and preventive properties of Petroselinum sativum seed extracts are well known, but the anticancer activity of alcoholic extracts and oil of Petroselinum sativum seeds on human breast cancer cells have not been explored so far. Therefore, the present study was designed to investigate the cytotoxic activities of these extracts against MCF-7 cells. Cells were exposed to 10 to $1000{\mu}g/ml$ of alcoholic seed extract (PSA) and seed oil (PSO) of Petroselinum sativum for 24 h. Post-treatment, percent cell viability was studied by 3-(4, 5-dimethylthiazol-2yl)-2, 5-biphenyl tetrazolium bromide (MTT) and neutral red uptake (NRU) assays, and cellular morphology by phase contrast inverted microscopy. The results showed that PSA and PSO significantly reduced cell viability, and altered the cellular morphology of MCF-7 cells in a concentration dependent manner. Concentrations of $50{\mu}g/ml$ and above of PSA and $100{\mu}g/ml$ and above of PSO were found to be cytotoxic in MCF-7 cells. Cell viability at 50, 100, 250, 500 and $1000{\mu}g/ml$ of PSA was recorded as 81%, 57%, 33%, 8% and 5%, respectively, whereas at 100, 250, 500, and $1000{\mu}g/ml$ of PSO values were 90%, 78%, 62%, and 8%, respectively by MTT assay. MCF-7 cells exposed to 250, 500 and $1000{\mu}g/ml$ of PSA and PSO lost their typical morphology and appeared smaller in size. The data revealed that the treatment with PSA and PSO of Petroselinum sativum induced cell death in MCF-7 cells.

Chemical Composition and Biological Activities of Elsholtzia ciliata (Thunb.) Hylander (향유의 향기성분 분석 및 생리활성 검정)

  • Jeong, Jae-Hoon;Lim, Heung-Bin
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.6
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    • pp.463-472
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    • 2004
  • This study was carried out to investigate the chemical composition of essential oils, absolutes and oleoresins isolated from Elsholtzia ciliata and the biological activities of them. Yields of essential oils, absolutes and oleoresins were 0.34%, 11.33% and 15.24%, respectively. The major component was naginate ketone in essential oils, methyl linolenate in absolutes and 9,12,15-octadecatrienoic acid in oleoresins. Eseential oils and oleoresins showed the inhibitory activities in enzyme-dependent, enzyme-independent and autooxidatve lipid peroxidation systems. $EC_{50}$ values in neutral red uptake assays 24 h of exposure times were $23.3\;{\mu}g/m{\ell}$, $341.0\;{\mu}g/m{\ell}$ and $17.2\;{\mu}g/m{\ell}$ in essential oils, absolutes and oleoresins, respectively, and essential oils and oleoresins showed the cytotoxic effect at the only high dose. Absolutes and oleoresins did not show antibiotic and mutagenic activities. On the contrary, essential oils with over $500\;{\mu}g/plate$ showed antibiotic and mutagenic activities in Ames test. Essential oils and oleoresins have a prolongating effect the ciliostasis of rat trachea.