• Title/Summary/Keyword: Cellular System

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Evaluation of Antioxidant Activity of Sugar Alcohols Using TOSC (Total Oxy-radical Scavenging Capacity) Assay (TOSC 법을 이용한 당알코올의 항산화 활성 평가)

  • Kang, Keon-Wook;Kwak, Sang-Hoon;Yun, Sei-Young;Kim, Sang-Kyum
    • Toxicological Research
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    • v.23 no.2
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    • pp.143-150
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    • 2007
  • Although animal and epidemiological studies have suggested oxidative stress as an etiological factor in pathogenesis including cancer, inflammation, sepsis, fibrosis, cardiovascularlneurodegenerative diseases and aging-related disorders, conflicting results have been obtained in clinical trial with antioxidants. The reason for this discrepancy remains unknown but may be due, in part, to the lack of a validated assay system for evaluating antioxidant capacity. The antioxidant activity of a series of sugar alcohols against peroxyl radicals, hydroxyl radicals and peroxynitrites was determined by the total oxy-radical scavenging capacity (TOSC) assay and cell-based assay using H4IIE cells. Specific TOSC values calculated from the slope of the linear regression for erythritol, xylitol, sorbitol or mannitol against peroxyl radicals was $2.1{\pm}0.2,\;3.7{\pm}0.3,\;9.1{\pm}0.3$ or $8.7{\pm}1.1$ TOSC/mM, respectively. Specific TOSC values for erythritol, xylitol, sorbitol or mannitol against peroxynitrite was $1.9{\pm}0.3,\;3.9{\pm}0.4,\;7.8{\pm}0.7$ or $7.7{\pm}0.5$ TOSC/mM, respectively. These results suggest that oxy-radical scavenging capacity is dependent on the number of aliphatic hydroxyl group in sugar alcohols of monosaccharide. Tert-butylhydroperoxide (t-BHP)-induced cell toxicity determined by MTT assay was marginally attenuated by 10 mM erythritol, but completely inhibited by 10 mM xylitol, 2 mM sorbitol or 0.75 mM maltitol, a disaccharide alcohol. Oxidative stress markers, such as glutathione (GSH) and malondial-dehyde (MDA) levels, were measured in t-BHP-treated cells using HPLC equipped with a fluorescence detector and a reverse phase column. Erythritol did not change the levels of GSH and MDA in H411E cells treated with t-BHP. The t-BHP-induced changes in cellular GSH and MDA levels were ameliorated by 10 mM xylitol and completely blocked by 10 mM sorbitol and maltitol. These results indicate that sugar alcohols protect cells against oxidative stress via scavenging oxy-radical and suggest that TOSC assay in conjunction with cell-based assay is a valid method for evaluating antioxidant capacity of natural and synthetic chemicals.

A Study on Preparation and Binding Properties of Germanium-fortified Yeast (게르마늄강화효모의 제조 및 이의 게르마늄 결합에 관한 연구)

  • Lee, Sung-Hee;Ahn, Sang-Doo;Rho, Sook-Nyung;Sohn, Tsang-Uk
    • Applied Biological Chemistry
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    • v.48 no.4
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    • pp.382-387
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    • 2005
  • The aim of this study was to identify binding properties of germanium (Ge) in Germanium-fortified Yeast using optimum manufacturing process. The ratio of yeast cell and germanium solution was 1 : 0.5 (50%), and pH 6.5, $35^{\circ}C$ and 20 h during fermentation, and Germanium-fortified Yeast produced. In results of the XRD, NMR and FT-IR analysis, it was different adding inorganic Ge $(GeO_2)$ during fermentation process from transformed into germanium in Germanium-fortified Yeast. And germanium concentration was not shown any difference before and after in the dialysis test with SGF (simulated gastric fluids). Therefore, Germanium-fortified Yeast of Geranti made by using biosynthetic technology was considered that transformed into organic properties during fermentation process. And, this result showed that Germanium-fortified Yeast was not dissociated under SGF (simulated gastric fluids) condition because of its structural binding safety. Thus, Germanium-fortified Yeast was transformed into organic germanium during biosynthetic cultivation. It is expected that this Germanium-fortified Yeast can be applied as a new dietary functional materials for cellular immunity, recovery of injured cells and immune system, and possible anticancer activities by activation immune cells like macrophage.

The Effect of Vitamin A Derivatives on the Activity of Drug-metabolizing Enzyme in Rat Liver (Vitamin A 유도체로 인한 간의 약물대사효소 변동)

  • Lee, H.W.;Ryu, K.Z.;Ro, J.Y.;Hong, S.S.
    • The Korean Journal of Pharmacology
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    • v.18 no.1
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    • pp.65-72
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    • 1982
  • It has been known that retinoids are intrinsically of critical importance for control of premalignant epithelial cell differentiation. In the absence of retinoids, normal cellular differentiation and growth does not occur in epithelia such as those of trachea and bronchi. Furthermore, it was also reported that retinoid deficiency enhanced susceptibility to chemical carcinogenesis in the respiratory system, in the bladder, and in the colon of the experimental animal. In 1974, Bollag examined the effects of synthetic retinoids in prevention of development of cancer and demonstrated synthetic retinoids to have more favorable therapeutic index than retinoic acid for causing regression of skin papilloma in mice. Therefore, it was assumed that this anticarcinogenic effect of vitamin A derivatives could be due to modification of the metabolism of the carcinogenic polycyclic hydrocarbon, which must first be activated to exert their effect. Hill and Shih reported that vitamin A compounds and analogs had inhibitory effect on drug metabolizing enzyme from liver and lung tissue of mouse and hamster. Lucy suggested that the chemoprevention effect of vitamin A derivatives is due to reaction with molecular oxygen, and it is possible that inhibition of hydroxybenzpyrene formation is a result of this property. On the other hand, butylated hydroxytoluene which is a potent antioxidant strongly inhibited the formation of mammary tumor induced by dimethylbenranthracene. Also, it was observed that this antioxidant inhibited cancer induction in rats by N-2-fluo-renylacetamide. The purpose of this experiment was to investigate the effect of vitamin A derivatives such as retinoic acid and retinoid on drug-metabolizing enzyme and to determine whether riboflavin tetrabutylate or vitamin E could prevent of modify any changes induced by vitamin A delivatives in the rats. The results obtained were as followings. 1) Body weight was significantly reduced by retinoic acid, but not by retinoid. 2) Retinoic acid markedly increased liver weight while retincid showed no effect on liver weight. Treatment of riboflavin tetrabutylate did not affect retinoic acid-induced change in both body weight and liver weight. 3) Both retinoic acid and retinoid remarkably decreased the activity of aminopyrine demethylase. Pretreatment of riboflavin tetrabutylate, however, prevented inhibitory effect of retinoic acid on the enzyme activity. 4) No significant effect of vitamin E on aminopyrine demethylase was observed in both groups treated with retinoic acid and retinoid.

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Resveratrol-loaded Nanoparticles Induce Antioxidant Activity against Oxidative Stress

  • Kim, Jae-Hwan;Park, Eun-Young;Ha, Ho-Kyung;Jo, Chan-Mi;Lee, Won-Jae;Lee, Sung Sill;Kim, Jin Wook
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.2
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    • pp.288-298
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    • 2016
  • Resveratrol acts as a free radical scavenger and a potent antioxidant in the inhibition of numerous reactive oxygen species (ROS). The function of resveratrol and resveratrol-loaded nanoparticles in protecting human lung cancer cells (A549) against hydrogen peroxide was investigated in this study. The 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay was performed to evaluate the antioxidant properties. Resveratrol had substantially high antioxidant capacity (trolox equivalent antioxidant capacity value) compared to trolox and vitamin E since the concentration of resveratrol was more than $50{\mu}M$. Nanoparticles prepared from ${\beta}$-lactoglobulin (${\beta}$-lg) were successfully developed. The ${\beta}$-lg nanoparticle showed 60 to 146 nm diameter in size with negatively charged surface. Non-cytotoxicity was observed in Caco-2 cells treated with ${\beta}$-lg nanoparticles. Fluorescein isothiocynate-conjugated ${\beta}$-lg nanoparticles were identified into the cell membrane of Caco-2 cells, indicating that nanoparticles can be used as a delivery system. Hydrogen peroxide caused accumulation of ROS in a dose- and time-dependent manner. Resveratrol-loaded nanoparticles restored $H_2O_2$-induced ROS levels by induction of cellular uptake of resveratrol in A549 cells. Furthermore, resveratrol activated nuclear factor erythroid 2-related factor 2-Kelch ECH associating protein 1 (Nrf2-Keap1) signaling in A549 cells, thereby accumulation of Nrf2 abundance, as demonstrated by western blotting approach. Overall, these results may have implications for improvement of oxidative stress in treatment with nanoparticles as a biodegradable and non-toxic delivery carrier of bioactive compounds.

고온성 알콜발효 효모의 Alcohol Dehydrogenase의 특성

  • Yea, Sang-Soo;Lim, Si-Kyu;Sohn, Ho-Yong;Jin, Ing-Nyul;Rhee, In-Koo;Kim, Young-Ho;Seu, Jung-Hwn;Park, Wan
    • Microbiology and Biotechnology Letters
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    • v.25 no.4
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    • pp.386-390
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    • 1997
  • The characteristics of alcohol dehydrogenase (ADH, EC 1.1.1.1, alcohol:NAD oxidoreductase) of thermotolerant alcohol-producing yeasts, Saccharomyces cerevisiae RA-74-2 and Kluyveromyces marxianus RA-912, were compared with that of mesophilic S. cerevisiae D, an industrial strain. Under anaerobic culture condition, both S. cerevisiae RA-74-2 and D had similar level of ADH activity at 30$\circ$C, and the activity of S. cerevisiae RA-74-2 at 37$\circ$C was the same level at 30$\circ$C. However, the level of ADH activity of S. cerevisiae D at 37$\circ$C decreased about 70% of that at 30$\circ$C. The level of enzyme activity of K. marxianus RA-912, which showed lower alcohol productivity than S. cerevisiae RA-74-2 and D, was about 43% of those strains at 30$\circ$C, and decreased somewhat at 37$\circ$C. The results showed a good correlation between the alcohol productivities and the level of ADH activities of these strains grown at 30$\circ$C and 37$\circ$C. And the higher heat stability of ADH of S. cerevisiae RA-74-2 than that of S. cerevisiae D seemed to reflect the ability of high temperature fermentation. Despite of its fermentation ability even at 45$\circ$C, however, the ADH of K. marxianus RA-912 showed lower heat stability than that of S. cerevisiae D. Both S. cerevisiae RA-74-2 and D showed similar patterns of two bands of ADH isozyme, and the low band of S. cerevisiae RA-74-2 moved slightly faster than that of S. cerevisiae D. The staining intensity of the bands of S. cerevisiae D at 37$\circ$C was weaker than those at 30$\circ$C. However, S. cerevisiae RA-74-2 showed no differences in total intensity of the bands of 30$\circ$C and 37$\circ$C. As the patterns of cellular proteins and ADH isozyme of K. marxianus RA-912 were different from S. cerevisiae RA-74-2 and D, K. marxianus might have its own characteristic ADH system.

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A Study on the Causes That Have Influence over the Effect of PPL in the Game (게임 속 PPL의 효과에 영향을 미치는 요인에 관한 연구)

  • Kim, Young-Rak;Cho, Youn-Gon;Choi, Gui-Young
    • Journal of Korea Multimedia Society
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    • v.13 no.8
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    • pp.1256-1262
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    • 2010
  • The game has been rapidly evolving through various media such as computer, console game machine, cellular phone and PMP based on the advanced development of scientific technology. In terms of demand, the interest in and desire to consume the game as a way of spending people's spare time have been on the increase constantly while the level of income has been improved. Eventually, the game has gradually expanded its scope of supply and demand, has established its own status as one of the media that is scientifically-intensive and has been developed into a game industry, a large-scale industry. Unlike image media, the methods of exposure in PPL are varied in accordance with the genre of games. This study divides the causes that have influence over the effect of PPL in the game into the genre of game and the skill of gamer. The results of the experiment on how much the aforementioned two elements have influence over the effect of PPL are as in the following: It has been demonstrated that the effect of PPL could appear different according to the genre of game and the skill of gamer on the game. Besides, the genre of game that is dynamic in its screen change in the game has relatively lower effect of PPL than that is not dynamic. Meanwhile, the persons who are highly skilled in the game have higher degree of recognition and preference to the inserted PPL than those who are lowly skilled. In this regard, it has given us a theoretical ground that the fees system for PPL ads should be established variously in accordance with the genre of game and the level of online game users.

Mutant p53-Notch1 Signaling Axis Is Involved in Curcumin-Induced Apoptosis of Breast Cancer Cells

  • Bae, Yun-Hee;Ryu, Jong Hyo;Park, Hyun-Joo;Kim, Kwang Rok;Wee, Hee-Jun;Lee, Ok-Hee;Jang, Hye-Ock;Bae, Moon-Kyoung;Kim, Kyu-Won;Bae, Soo-Kyung
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.4
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    • pp.291-297
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    • 2013
  • Notch1 has been reported to be highly expressed in triple-negative and other subtypes of breast cancer. Mutant p53 (R280K) is overexpressed in MDA-MB-231 triple-negative human breast cancer cells. The present study aimed to determine whether the mutant p53 can be a potent transcriptional activator of the Notch1 in MDA-MB-231 cells, and explore the role of this mutant p53-Notch1 axis in curcumin-induced apoptosis. We found that curcumin treatment resulted in an induction of apoptosis in MDA-MB-231 cells, together with downregulation of Notch1 and its downstream target, Hes1. This reduction in Notch1 expression was determined to be due to the decreased activity of endogenous mutant p53. We confirmed the suppressive effect of curcumin on Notch1 transcription by performing a Notch1 promoter-driven reporter assay and identified a putative p53-binding site in the Notch1 promoter by EMSA and chromatin immunoprecipitation analysis. Overexpression of mutant p53 increased Notch1 promoter activity, whereas knockdown of mutant p53 by small interfering RNA suppressed Notch1 expression, leading to the induction of cellular apoptosis. Moreover, curcumin-induced apoptosis was further enhanced by the knockdown of Notch1 or mutant p53, but it was decreased by the overexpression of active Notch1. Taken together, our results demonstrate, for the first time, that Notch1 is a transcriptional target of mutant p53 in breast cancer cells and suggest that the targeting of mutant p53 and/or Notch1 may be combined with a chemotherapeutic strategy to improve the response of breast cancer cells to curcumin.

High Throughput-compatible Screening of Anti-oxidative Substances by Insect Extract Library (약용곤충추출물 라이브러리를 이용한 항산화 활성의 초고속 검색)

  • Park, Ja-Young;Heo, Jin-Chul;An, Sang-Mi;Yun, Eun-Young;Han, Sang-Mi;Hwang, Jae-Sam;Kang, Seok-Woo;Yun, Chi-Young;Lee, Sang-Han
    • Food Science and Preservation
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    • v.12 no.5
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    • pp.482-488
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    • 2005
  • Oxidant stress is well-known for a pivotal parameter related to neuro-inflammatory diseases including Alzheimer's disease, Parkinson's disease, and ALS (Lou Gehrig's disease). In order to effectively screen for anti-inflammatory agents, we first established the infrastructure of high throughput screening for anti-oxidant agents from medicinal insect library extracted with water, methanol, ethanol, and dimethyl sulfoxide. By the screening system, we found that Tenodera angustipennis Saussure, Pyrocoela rupa Olivier and Papilio maackii Mntris had strong anti-oxidant activity. Moreover, Tenodera angustipennis Saussure and Tenodera aridifolia (Stoll) exhibited protection effects of cellular damage by treatment of an oxidant hydrogen peroxide. Together, the results suggest that some selected hits could be a potential agent against neuro-inflammation, although the in vivo studies should be clearly tested.

Insect Resistance of Tobacco Plant Expressing CpBV-ELP1 Derived from a Polydnavirus (폴리드나바이러스 유래 CpBV-ELP1 발현 담배의 내충성)

  • Kim, Eunseong;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.56 no.1
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    • pp.19-28
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    • 2017
  • Polydnaviruses (PDVs) are a group of double-stranded DNA viruses symbiotic to some endoparasitoid wasps. Cotesia plutellae bracovirus (CpBV) is a PDV symbiotic to an endoparasitoid wasp, C. plutellae, parasitizing young larvae of Plutella xylostella. An early expressed gene, CpBV-ELP1, plays an important role in the parasitism by suppressing host cellular immunity by its cytotoxic activity against hemocytes. This study aimed to test its oral toxicity against insect pest by expressing it in a recombinant tobacco plant. A recombinant CpBV-ELP1 protein was produced using a baculovirus expression system and secreted to cell culture medium. The cell cultured media were used to purify CpBV-ELP1 by a sequential array of purification steps: ammonium sulfate fractionation, size exclusion chromatography, and ion exchange chromatography. Purified rCpBV-ELP1 exhibited a significant cytotoxicity against Spodoptera exigua hemocytes. CpBV-ELP1 was highly toxic to the fifth instar larvae of S. exigua by injection to hemocoel. It also showed a significant oral toxicity to fifth instar larvae of S. exigua by a leaf-dipping assay. CpBV-ELP1 was cloned into pBI121 vector under CaMV 35S promoter with opaline synthase terminator. Resulting recombinant vector (pBI121-ELP1) was used to transform Agrobacterium tumefaciens LBA4404. The recombinant bacteria were then used to induce callus of a tobacco (Nicotiana tabacum Xanthi) leaves and subsequent generation (T1) plants were selected. T1 generation tobacco plants expressing CpBV-ELP1 gave significant insecticidal activities against S. exigua larvae. These results suggest that CpBV-ELP1 gene can be used to control insect pests by constructing transgenic crops.

Protective effects of Seoritae Chungkukjang added with green tea powder against 3-morpholinosydnonimine-induced oxidative stress (녹차 첨가 서리태 청국장의 3-morpholinosydnonimine에 의한 산화적 스트레스 개선 효과)

  • Cho, Eun-Ju;Park, Hyun-Young;Lee, Sang-Hyun;Kim, Hyun-Young
    • Korean Journal of Agricultural Science
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    • v.42 no.4
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    • pp.407-414
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    • 2015
  • To increase antioxidative activity of Chungkukjang, the protective effect of Seoritae Chungkukjang (SC) added with green tea powder against oxidative stress was evaluated under the cellular system using LLC-$PK_1$ cells. The treatment of 3-morpholinosydnonimine showed increase in lipid peroxidation, and decrease in endogenous anti-oxidant enzymes activity and cell viability. The methanol extract of SC inhibited lipid peroxidation by 70.9%, and significantly increased cell viability up to more than 33.2%. In addition, it enhanced superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities. Particularly, the addition of green tea in SC exerted protective effect against oxidative stress by ONOO- through elevation in activities of SOD and GSH-Px, and inhibition of lipid peroxidation. More addition of green tea showed stronger protective activity. These results suggest that the addition of green tea to SC leads to the increase in the antioxidative effect of Chungkukjang through elevation in antioxidative enzyme activities and protection from lipid peroxidation.