In this study, we investigated the antioxidant activities, inhibition activity against ACE (angiotensin converting enzyme) and antitumor activity of extract from Acanthopanax senticosus HARMS fruits for development novel functional resources. In order to understand the factors responsible for the potent antioxidant and antihypertensive ability of fruits in A. senticosus, it has been evaluated for anti-oxidative activity using Fenton's reagent/ethyl linoleate system and for free radical scavenging activity using the l,l-diphenyl-2-picryl hydrazyl free radical generating system. The fruits extract of A. senticosus showed higher antioxidant activities than positive control, ${\alpha}$-tocopherol at all concentrations, while fruits extract of A. senticosus showed same degree of radical scavenging activity with positive control, ${\alpha}$-tocopherol. The ability of fruit extracts from A. senticosus to influence the inhibitory activity of angiotensin converting enzyme (ACE) and xanthine oxidase (XOase) has also been discussed. The activity of growth-inhibitory of fruit extracts of A. senticosus was screened by SRB (sulphorhodamine B) method on diverse cancer cells representing different types of cancers. The fruit extracts of A. senticosus showed moderate inhibition on proliferation of LNCaP and MOLT-4F cells and did not inhibit the proliferation of other cancer cells. The fruit extracts of A. senticosus inhibited the proliferation of cancer cells with $GI_{50}$ values ranging from 5 to $10{\mu}g/mL$. This result revealed that the fruit extracts of A. senticosus was expected to be good candidate for development into source of free radical scavengers, antihypertentive, and anti-tumor agent.
Tributyltin (TBT) used world-wide in antifouling paints for ships is a widespread environmental pollutant and cause reproductive organs atrophy in rodents. At low doses, antiproliferative modes of action have been shown to be involved, whereas at higher doses apoptosis seems to be the mechanism of toxicity in reproductive organs by TBT. In this study, we investigated that the mechanisms underlying DNA fragmentation induced by TBT in the rat leyding cell line, R2C. Effects of TBT on intracellular Ca$\^$2+/ level and reactive oxygen species (ROS) were investigated in R2C cells by fluorescence detector. TBT significantly induced intracellular Ca$\^$2+/ level in a time-dependent manner. The rise in intracellular Ca$\^$2+/ level was followed by a time-dependent generation of reactive oxygen species (ROS) at the cytosol level. Simultaneously, TBT induced the release of cytochrome c from the mitochondrial membrane into the cytosol. Furthermore, ROS production and the release of cytochrome c were reduced by BAPTA, an intracellular Ca$\^$2+/ chelator, indicating the important role of Ca$\^$2+/ in R2C during these early intracellular events. In addition, Z-DEVD FMK, a caspase-3 inhibitor, decreased apoptosis by TBT. Taken together, the present results indicated that the apoptotic pathway by TBT might start with an increase in intracellular Ca$\^$2+/ level, continues with release of ROS and cytochrome c from mitochondria, activation of caspases,and finally results in DNA fragmentation.
Journal of the Korean Society of Food Science and Nutrition
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v.45
no.4
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pp.510-517
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2016
This study was conducted to investigate the antioxidant and hepatoprotective effects of eriodictyol compound against hydrogen peroxide-induced oxidative stress in HepG2 cells by measuring expression levels of antioxidant enzymes, liver function index enzyme activities, and inhibitory effects against reactive oxygen species (ROS) production. HepG2 cell viability was assessed using 3-(4,5-dimethyl thiazole-2-yl)-2,5-diphenyl tetrazolium bromide assay. In the concentration range of $10{\sim}50{\mu}g/mL$, eriodictyol displayed over 98% cell viability in HepG2 cells. The effects of increased gene expression on hydrogen peroxide-induced oxidative stress were analyzed by monitoring antioxidant enzyme (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPx) gene expression levels using real-time PCR. Eriodictyol compound significantly increased gene expression levels of SOD, CAT, and GPx in a dose-dependent manner ($10{\sim}50{\mu}g/mL$). Hepatoprotective effects against hydrogen peroxide-induced oxidative stress were analyzed by monitoring glutamic oxaloacetic transaminase (GOT), lactate dehydrogenase (LDH), and gamma-glutamyl transferase (GGT) activities in HepG2 cell culture medium using a biochemistry analyzer. Eriodictyol compound significantly reduced GOT, LDH, and GGT activities in a dose-dependent manner in HepG2 cells. ROS level in HepG2 cells was analyzed by 2',7'-dichlorofluorescein fluorescence diacetate assay, and eriodictyol compound effectively reduced the intracellular ROS level in HepG2 cells. The results reveal that eriodictyol compound can be useful for development of effective antioxidant and hepatoprotective agents.
Purpose: A retrospective study was performed to evaluate the efficiency and feasibility of twice daily radiation therapy plus concurrent chemotherapy for limited-stage small cell lung cancer in terms of treatment response, survival, patterns of failure, and acute toxicities. Materials and Methods: Between February 1993 and October 2002, 76 patients of histologically proven limited-stage small cell lung cancer (LS-SCLC) were treated with twice daily radiation therapy and concurrent chemotherapy. Male was in 84% (64/76), and median age was 57 years (range, 32-75 years). Thoracic radiation therapy consisted of 120 or 150 cGy per fraction, twice a day at least 6 hours apart, 5 days a week. Median total dose was 50.4 Gy (range, 45-51 Gy). Concurrent chemotherapy consisted of CAV ($cytoxan\;1000mg/m^2,\;adriamycin\;40mg/m^2,\;vincristine\;1mg/m^2$) alternating with PE ($cisplatin\;60mg/m^2,\;etoposide\;100mg/m^2$) or PE alone, every 3 weeks. The median cycle of chemotherapy was six (range, 1-9 cycle). Prophylactic cranial irradiation (PCI) was recommended to the patients who achieved a complete response (CR). PCI scheme was 25 Gy/10 fractions. Median follow up was 18 months (range, 1-136 months). Results: Overall response rate was 86%; complete response in 39 (52%) and partial response in 26 (34%) patients. The median overall survival was 23 months. One, two, and three year overall survival rate was 72%, 50% and 30%, respectively. In univariate analysis, the treatment response was revealed as a significant favorable prognostic factor for survival (p<0.001). Grade 3 or worse acute toxicities were leukopenia in 46 (61%), anemia in 5 (6%), thrombocytopenia in 10 (13%), esophagitis in 5 (6%), and pulmonary toxicity in 2 (2%) patients. Of 73 evaluable patients, 40 (55%) patients subsequently had disease progression. The most frequent first site of distant metastasis was brain. Conclusion: Twice daily radiation therapy plus concurrent chemotherapy produced favorable response and survival for LS-SCLC patients with tolerable toxicities. To improve the treatment response, which proved as a significant prognostic factor for survival, there should be further investigations about fractionation scheme, chemotherapy regimens and compatible chemoradiotherapy schedule.
Background : Arsenic trioxide ($As_2O_3$) has been used to treat acute promyelocytic leukemia, and it induces apoptosis in a variety of solid tumor cell lines including non-small cell lung cancer cells. However, nonsteroidal antiinflammatory drugs (NSAID) can enhance tumor response to chemotherapeutic drugs or radiation. It was previously demonstrated that a combination treatment with $As_2O_3$ and sulindac induces the apoptosis of NCI-H157 human lung carcinoma cells by activating the caspase cascade. This study aimed to determine if a combination treatment augmented its apoptotic potential through other pathways except for the activation of the caspase cascade. Material and Methods : The NCI-H157 cells were treated with $As_2O_3$, sulindac and antioxidants such as glutathione (GSH) and N-acetylcysteine (NAC). The cell viability was measured by a MTT assay, and the level of intracellular hydrogen peroxide ($H_2O_2$) generation was monitored fluorimetrically using a scopoletin-horse radish peroxidase (HRP) assay. Western blotting and mitochondrial membrane potential transition analysis were performed in order to define the mechanical basis of apoptosis. Results : The viability of the cells was decreased by a combination treatment of $As_2O_3$ and sulindac, and the cells were protected using antioxidants in a dose-dependent manner. The increased $H_2O_2$ generation by the combination treatment was inhibited by antioxidants. The combination treatment induced changes in the mitochondrial transmembrane potential as well as the expression of the Bcl-2 family proteins, and increased cytochrome c release into the cytosol. However, the antioxidants inhibited the effects of the combination treatment. Conclusion : Combination treatment with $As_2O_3$ and sulindac induces apoptosis in NCI-H157 human lung carcinoma cells via ROS generation with a mitochondrial dysfunction.
The heavy metal accumulations of avian eggshells were studied in order to test a feral pigeon (Columba livia)'s eggshell as an indicator for the environmental monitoring of pollutants. The reviews on the eggs of the different 19 avian species showed that it is the eggshell rather than the egg content that can better reflect the heavy metals in the environment; in most cases the CVs (coefficients of variations) of the heavy metal concentrations in the eggshells were higher than those in the egg contents. This can indicate that the heavy metal accumulations are homeostatically controlled in the egg contents, but the accumulations in the eggshells are varied according to the environmental conditions. To test the reviews, the feral pigeon eggs from the two different sites, one representing urban and the other rural environment, were analyzed for lead (Pb) and cadmium (Cd). The result showed that the eggshells of the urban pigeons (Hangang) had the higher metal concentrations than those of the rural pigeons (Hampyeong). The same difference can also be found in the internal organs (liver, bone) and blood. However, the analyses of the egg contents between the two sites did not reveal the differences. In other words, the result suggests that the feral pigeons, like the other avian species, are able to control the heavy metals into the egg contents homeostatically. Therefore, it is more useful to use the feral pigeon eggshell rather than the egg content in case of monitoring heavy metals in different habitats.
The aim of research is the evaluation of the $Cr^{6+}$ emission features of the liquid injection through emission experiments in varying conditions, based on a field-mixing ratio. The results showed that the content of $Cr^{6+}$ content in cement measured had an Ordinary Potland Cement (OPC) of 25.3 mg/kg, which constitute the largest portion among the other materials. Likewise, the emission experiment of homo-gel and sand-gel generally satisfied the standard of KSLT (Korea Standard Leaching Test) in waste of 1.5 mg/L, but in case of the standard of KSLT in soil the emission of OPC $Cr^{6+}$ of 4.85 mg/kg. These conditions is a little exceeded the criteria in the ‘Ga’ area in terms of Korea Soil Environmental Preservation Law. In addition, results generated by the mock-up injection facilities revealed that $Cr^{6+}$ emission increased as Water/Cement and injection pressure increased. At injection pressure higher than 4 kg/㎤, $Cr^{6+}$ emission exceeded the water preservation standard of 0.5 mg/L. Similarly, a pattern experiment of C $r^{6+}$ emission according to pH was conducted, in order to evaluate the $Cr^{6+}$ emission features of grout materials in leachate below pH 5 such as pH 4 acid rain or landfill. Results show that $Cr^{6+}$ emission dramatically increased in high acidic or basic state. It indicates that $Cr^{6+}$ emission will probably increase in an environment where grout materials are injected. On the other hand, concentration of leachate was determined in areas where grout materials are used. The results show that the concentration of emission in an ultra purity condition does not manifest intensity, and is affected in the OPC>MC>SC order. It means that the pollutants or $Cr^{6+}$ emission increases with decreasing concentration. As such, $Cr^{6+}$ emission will probably exceed the countermeasure criteria according to the types of gout materials. Similarly, high pressure or injection will cause increased $Cr^{6+}$ emission. Therefore, the selection of materials or mixing ratio should be considered in general as well as according to specific industries, based on the strength and pH of $Cr^{6+}$ emission.
Ethanol is a very valuable material, however, it is also a source of stress, as the accumulation of ethanol in a medium inhibits cell viability and decreases productivity of the target product. Therefore, the ethanol tolerance of yeast, which is closely related to ethanol productivity, is an important factor in industrial ethanol production. In this study, the YDJ1 and PEP5 genes were selected as target genes for elucidating ethanol-tolerant mechanisms by analyzing the expression regulation of these genes. The pA-YDJ1 and pA-PEP5 plasmids containing YDJ1 and PEP5 genes under an ADH1 promoter, respectively, were constructed and transformed into BY4742 (host strain), BY4742△ydj1, and BY4742△pep5 strains. The ethanol tolerance in the BY4742△ydj1/ pA-YDJ1 and BY4742△pep5/pA-PEP5 transformants was restored by overexpression of the YDJ1 and PEP5 genes to the host strain level. The YDJ1 and PEP5 genes were also introduced into the double gene disruptant (BY4742△ydj1△pep5) to investigate the expression regulation of the YDJ1 and PEP5 genes. The simultaneous overexpression of the YDJ1 and PEP5 genes restored ethanol tolerance to the 90% level of the BY4742 strain under 8% ethanol stress. The YDJ1 gene induced more overexpression of the PEP5 gene in the BY4742△ydj1 △pep5/pA-YDJ1, pA-PEP5 strain, suggesting that the YDJ1 gene partially regulates the expression of the PEP5 gene as an upstream regulator.
Journal of the Society of Cosmetic Scientists of Korea
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v.43
no.2
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pp.157-164
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2017
Stevia (Stevia rebaudiana) is a perennial plant of the genus Stevia, originated in South America. It stores many forms of glycosides, mainly stevioside and rebaudioside A, in which steviol is the basic structure. Steviol glycosides, widely used as sweeteners, are superior to sugar in sweetness. Recently, it has been reported that steviol glycosides are involved not only in the skin whitening and anti-inflammatory effect but also in enhancing skin barrier function through tight junction regulation. Thus, we examined anti-inflammatory effect of rebaudioside A and tried to identify its potential for improving atopic dermatitis as cosmetic ingredients. To investigate the anti-inflammatory effect, cell viability and mRNA expression level of inflammation-related cytokines were measured using mouse macrophage RAW264.7 cells. Cell counting kit 8 (CCK-8) assay was carried out to measure cell viability and the maximum concentration without cytotoxicity was set to $250{\mu}M$. A quantitative real-time RT-PCR method was used for the study of the inflammatory suppression of rebaudioside A. Rebaudioside A inhibited expression of inducible nitric oxide synthase (iNOS) up to 47% and COX-2 up to 41% compared to LPS treated condition. NO synthesis was decreased by rebaudioside A. Also, mRNA expression of interleukin (IL)-$1{\alpha}$, IL-$1{\beta}$ and IL-6 in LPS-stimulated RAW264.7 cells was decreased to 40%, 45% and 59%, respectively, as a concentration-dependent manner. In conclusion, rebaudioside A inhibited the inflammatory response by regulation of cytokine gene expression. From these results, we expect that steviol glycoside, such as rebaudioside A, can be used as a material for improving atopic dermatitis in the future.
This study examined a new functional cosmetic material possessing application possibility of Sambucus sieboldiana var. pendula (Nakai) (SS) extract. For this, we analyzed the toxic effect of the SS extract on macrophages (RAW 264.7 cells) by performing MTT assay. Results of the MTT assay showed ${\geq}100%$ cell viability after treatment with $500{\mu}g/ml$ SS extract. To determine the anti-inflammatory activity of the SS extract, we examined its inhibitory effect on lipopolysaccharide (LPS)-induced NO production in RAW 264.7 cells by performing Griess assay. Result of the Griess assay showed that the SS extract inhibited LPS-induced NO production in a concentration-dependent manner. Next, we examined the effect of the SS extract on the production of proinflammatory factors inducible NOS (iNOS) and cyclooxygenase-2 (COX-2) in LPS-stimulated RAW 264.7 cells. First, we determined the inhibitory effect of 50, 100, and $500{\mu}g/ml$ SS extract on iNOS and COX-2 protein expression by performing western blot analysis, with ${\beta}$-actin as a positive control. Results of western blotting showed that treatment with $500{\mu}g/ml$ SS extract decreased iNOS and COX-2 protein expression by 31.2% and 54.7%, respectively. Next, we determined the inhibitory effect of 50, 100, and $500{\mu}g/ml$ SS extract on iNOS and COX-2 mRNA expression by performing reverse transcription-polymerase chain reaction (PCR), with GAPDH as a positive control. Results of reverse transcription-PCR showed that treatment with $500{\mu}g/ml$ SS extract decreased the mRNA expression of iNOS and COX-2 by 72.2% and 89%, respectively. These results suggest that the SS extract is a highly valuable natural compound because of its functional components and anti-inflammatory activity.
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