• Title/Summary/Keyword: in vitro toxicity

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Synergistic action of pesticide mixtures using glutathione-s-transferase- and esterase-inhibiting properties in diamondback moth (Plutella xylostella L.) (Glutathione-S-transferase와 esterase 효소 저해특성을 이용한 농약의 혼합 상승효과)

  • Yu, Yong-Man;Hong, S.S.;Kim, S.;Hur, J.H.
    • The Korean Journal of Pesticide Science
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    • v.7 no.1
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    • pp.38-44
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    • 2003
  • In vitro inhibitory activity of 34 insecticides and 31 fungicides to glutathione-S-transferase and esterases extracted from rats was determined. Of tested pesticides, the pesticides with high activity on both detoxifying enzymes were mixed with pesticides that are known to be detoxified by detoxifying enzymes. Glutathione-S-transferase was inhibited by thiodicarb $(I_{50}:1.87\times10^{-4}M)$, thiocyclam $(7.40\times10^{-4}M)$, dithianon $(7.55\times10^{-5}M)$, and tolylfluanide $(8.66\times10^{-5}M)$, while esterases by dichlorvos $(8.95\times10^{-8}M)$, pirimicarb $(2.74\times10^{-6}M)$, pyrazophos $(3.31\times10^{-5}M)$, and benomyl $(4.96\times10^{-5}M)$. After acephate known to be detoxified by glutathione-S-transferase was mixed with glutathione-S-transferase-inhibiting pesticides and phenthoate known to be detoxified by esterases was mixed with esterases-inhibiting pesticides, insecticidal activities of such mixtures were determined against diamondback moth (PlutelLa xylostella L.). Synergistic action was observed in all pesticide combinations. The highest synergistic action was obtained when phenthoate was combined with dichlorvos, showing that co-toxicity coefficients were 1512 and 1877 after 24 and 48 hours of treatment, respectively. Several other combinations of pesticides, such as phenthoate with benomyl, and acephate with dithianon, also showed synergism, showing that their co-toxicity coefficients were about 1,000 and 500, after 24 hours of treatment, respectively. Our results showed that combinations of pesticides inhibited by detoxifying enzymes and ones detoxified by detoxifying enzymes resulted in increased toxicities of pesticides, suggesting that such combinations could be used to develop pesticide mixtures with more broad spectrum and high effectiveness.

In vitro Antimutagenic and Genotoxic Effects of Azadirachta indica Extract (님추출물의 in vitro 항돌연변이원성 및 유전독성 영향)

  • Yoon, Hyunjoo;Cho, Hyeon-Jo;Kim, Jin Hyo;Park, Kyung-Hun;Gil, Geun-Hwan;Oh, Jin-Ah;Cho, Namjun;Paik, Min-Kyoung
    • Journal of Applied Biological Chemistry
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    • v.57 no.3
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    • pp.219-225
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    • 2014
  • Azadirachta indica extract (AIE) has been regarded as a promising source of environment-friendly organic materials owing to their low mammalian toxicity. However, quite a bit of research has been reported that AIE may cause clastogens in human lymphocytes. Therefore, this study was conducted to evaluate the antimutagenic and genotoxicity of two samples of AIE. Antimutagenic test was experimented by using bacterial reverse mutation test. In the bacterial reverse mutation test, five strains Salmonella Typhimurim of two samples of AIE in order to evaluate its mutagenic potential. Bacterial reverse mutation test was also performed on positive control and negative control groups in the presence of the metabolic activation system (S-9 mix) and metabolic non-activation system. In the chromosome aberration test, Chinese hamster lung cells were exposed to AIE for 6 or 24 h with BPS, or for 6 h with S-9 mix. Negative and positive control groups were experimented for chromosome aberration test. As a result, the number of mutated colonies induced by 4-NQO were reduced by AIE treatment in all strains, indicating that AIE may have antimutagenic effects. Bacterial reverse mutation and chromosomal aberration were not shown at all concentration of AIE, regardless of activation of the metabolic system. we concluded that two AIE samples used in this study have no genotoxic effects to human, according to the genotoxicity battery system suggested by ICH (International Conference on Harmonization).

Anti-inflammatory Effect of Ethanol Extract from Sargassum fulvellum on Lipopolysaccharide Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears (LPS로 유도된 RAW 264.7 세포와 마우스 귀조직에 대한 참모자반 (Sargassum fulvellum) 에탄올 추출물의 항염증 효과)

  • Jeong, Da-Hyun;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Kang, Bo-Kyeong;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Ahn, Na-Kyung;Choi, Yeon-Uk;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.8
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    • pp.1158-1165
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    • 2014
  • This study investigated the anti-inflammatory effects of Sargassum fulvellum ethanol extract (SFEE) on the lipopolysaccharide (LPS)-induced inflammatory response. SFEE remarkably suppressed production of NO and pro-inflammatory cytokines (IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ at 50 and $100{\mu}g/mL$. There were no cytotoxic effects on proliferation of macrophages treated with SFEE compared to the control. SFEE reduced expression of iNOS and COX-2 proteins in a dose-dependent manner. The formation of edema in mouse ears was reduced at the highest dose tested compared to the control. Moreover, in the acute toxicity test, no mortality occurred in mice administered 5,000 mg/kg body weight of SFEE over the 2-week observation period. These results suggest that SFEE may have significant effects on inflammatory factors and be a potential anti-inflammatory therapeutic material.

Effect of Plant Extracts with Superoxide Dismutase-like Activity on Survival of Fruit Flies under Oxidative Stress (Superoxide Dismutase유사활성을 지닌 식물체가 Oxidative Stress를 받고 있는 초파리의 수명에 미치는 영향)

  • Han, Dae-Seok;Kwak, Jae-Hyock;Kim, Sang-Hee;Kim, Seok-Joong
    • Korean Journal of Food Science and Technology
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    • v.28 no.5
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    • pp.865-869
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    • 1996
  • Biological effect of aqueous extracts of 12 plants which showed superoxide dismutase (SOD)-like activity in vitro was evaluated using Drosophila melanogaster. Survival percentage of the flies was a criterion of effect when the flies were exported to paraquat, which generated superoxide anion radical in vivo. When flies were co-administered with paraquat and aqueous extracts of garlic, lettuce, kiwi, and nameko, they showed no defensive effect against of oxygen toxicity. If flies were exposed to 60 mM paraquat after adaptation to feed containing plant extracts with SOD-like activity for 10 days, however, survival percentage of flies fed with phytochemicals was $35{\sim}63%$ while that percentage of flies fed without phytochemical was only 11%. This result indicated that adaptation of flies to plant extracts with SOD-like activity could prevent the flies from oxidative injury. On the other hand, lettuce, kiwi, nameko, onion, persimmon, fern brake and cauliflower showed a reparative effect on an oxidative stress. Dropwort, shiitake, agaric mushroom and broccoli did not show such an effect.

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Screening of Traditional Herbal Medicines to Develop New Materials for Anti-aging and Anti-wrinkle in the Skin (전통 생약재에서의 항노화와 주름 개선 활성 신소재 탐색)

  • Lee Sang Eun;Son Dong Wook;Yoon Year Pill;Lim Tae Young;Cho Ja Wun;Kim Haen Su
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.2 s.51
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    • pp.147-152
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    • 2005
  • As a part of searching tot the natural components which inhibit the skin aging and wrinkle formation, the $80\%$ methanolie extracts of 121 species of traditional herbal medicines used to treat lung and skin disease were investigated for their in vitro anti-oxidative activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide radicals, and inhibitory activity against elastase. We selected 9 kinds of the traditional herbal medicines showing inhibitory activities of winkle formation. The effective concentrations of 9 candidates for anti-wrinke/skin firming activity was less than 0.1 mg/mL, and there is no toxicity to cell viability at these concentrations. Through analysis of human skin primary patch test data, the traditional herbal medicines represented non-irritant materials. We suggest that these 9 candidates with ability to help anti-wrinkle/skin firming may be useful for functional cosmetic materials.

Effects of Caffeine and calcium on the activities of the mouse osteoblastic cells (카페인과 칼슘이 골모 세포의 활성에 미치는 영향)

  • Chun, Youn-Sic;Baik, Hye-Jung
    • The korean journal of orthodontics
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    • v.32 no.2 s.91
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    • pp.129-142
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    • 2002
  • The purpose of this study was to evaluate the effects of caffeine and calcium on the activities of the osteoblastic cell from mouse calvaria. The author cultured osteoblastic cells obtained from the mouse calvaria and were divided into three groups : the caffeine-treated, the calcium-treated and the combine-treated group. In caffeine-treated group, the cell toxicity was measured by MTT assay at 1, 2 and 4 days after treatment of caffeine. In all groups, the densities of the mineralized bone nodules were measured by imaging analyzer after Von Kossa staining. The alkaline phosphotase (ALP) activities were measured at 2, 7, 14, 21 and 28 days and the interleukin-1 ${\beta}$ activities at 48 hours after treatment of caffeine and calcium. The measurements were statistically executed with ANOVA test and the results were as follows. 1. The cellular toxicity of the caffeine increased with the concentration of caffeine during the incubation period. 2. The maximum densities of mineralization were observed at 0.2 mM caffeine-treated group, 1.2 mM calcium-treated group, 0.1 mM caffeine and 1.8 mM calcium-treated group. 3. The activities of ALP were peaked at 14 days at calcium-treated group as no-treated. But, the activities of ALP increased with concentrations of caffeine at caffeine-treated group. At combine-treated group, the act of ALP were peaked at 24 days at 1.2 mM, 1.8 mM calcium-treated group, But decreased at 2.5 mM calcium-treated group. 4. The activites of the IL-1 ${\beta}$ were increased significantly at 0.2 mM caffeine-treated group, 1.8 mM calcium-treated group and 0.1 mM caffeine and 1.8 mM calcium-treated group. But, they were decreased at all groups of high concentration.

Sister Chromatid Exchange (SCE) Frequency and In Vitro Development of Mouse Zygote Cryopreserved by Vitrification (초자화 동결에 의한 생쥐 1-세포기배의 체외 발달과 SCE 빈도)

  • Kim, M.K.;Baik, C.S.;Uhm, S.J.;Kim, E.Y.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.3
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    • pp.379-384
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    • 1996
  • This study was undertaken to investigate the sister chromatid exchange (SCE) frequency and embryonic development after exposure to cryoprotectants and vitrification of mouse zygotes. Mouse IVF zygotes were cryopreserved by vitrification using vitrification solution, EFS40 (40% ethylene glycol, 30% Ficoll and 0.3 M sucrose in phosphate buffer saline containing 10% FBS). After mouse zygotes were exposed to EFS40 at $25^{\circ}C$ for 30 sec., they were immediately plunged into $LN_2$ or cultured for cryoprotectant toxicity test without freezing. The results obtained in these experiments were summarized as follows: After thawing, survival rates to the 2-cell stage of zygotes exposed to or vitrified in EFS 40 (98.5%, 95.2%) were not significantly difference compared with that of control (100%). However, the developmental rates upto blastocyst and hatching blastocyst in vitrified groups (66.7, 50.0%) were lower than those of control (93.9, 81.8%) or exposed group (94.0, 78.8%) (p<0.05). When the influence of vitrification and exposure to cryoprotectant on the in vitro development of mouse zygotes was assessed by the SCE frequency, the SCE frequency in exposed ($20.2{\pm}2.1$) to or vitrified embryos ($21.4{\pm}3.2$) was higher than that in control embryos ($16.8{\pm}1.5$). These results suggest that the frequency of SCE was increased after cryoprotectant exposure or Vitrification although developmental rates of zygotes upto blastocysts and /or hatching blastocysts were not afected by cryoprotectant.

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The Anticancer Effect and Mechanism of Photodynamic Therapy Using 9-Hydroxypheophorbide-a and 660 nm Diode Laser on Human Squamous Carcinoma Cell Line. (9-hydroxypheophorbide-a와 660 nm 다이오드 레이저를 이용한 광역학치료의 항암효과와 치료기전에 대한 연구)

  • Ahn, Jin-Chul
    • Journal of Life Science
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    • v.19 no.6
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    • pp.770-780
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    • 2009
  • A new photosensitizer, 9-Hydroxypheophorbide-a (9-HpbD-a), was derived from Spirulina platensis. We conducted a series of experiments, in vitro and in vivo, to evaluate the anticancer effect and mechanism of photodynamic therapy using 9-HpbD-a and 660 nm diode lasers on a squamous carcinoma cell line. We studied the cytotoxic effects of pheophytin-a, 9-HpbD-a, 9-HpbD-a red and 660 nm diode lasers in a human head and neck cancer cell line (SNU-1041). Cell growth inhibition was determined by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. The effects of 9-HpbD was higher than those of 9-HpbD-a red or pheophytin-a in PDT. We then tested the cytotoxic effects of 9-hydroxypheophorbide-a (9-HpbD-a) in vitro. The cultured SNU-I041 cells were treated with serial concentrations of 9-HpbD-a followed by various energy doses (0, 0.1, 0.5, 3.2 J/$cm^{2}$) and by various interval times (0, 3, 6, 9, 12 hr) until laser irradiation, then MTT assay was applied to measure the relative inhibitory effects of photodynamic therapy (PDT). Optimal laser irradiation time was 30 minutes and the cytotoxic effects according to incubation time after 9-HpbD-a treatment increased until 6 hours, after which it then showed no increase. To observe the cell death mechanism after PDT, SUN-I041 cells were stained by Hoechst 33342 and propidium iodide after PDT, and observed under transmission electron microscopy (TEM). The principal mechanism of PDT at a low dose of 9-HpbD-a was apoptosis, and at a high dose of 9-HpbD-a it was necrosis. PDT effects were also observed in a xenografted nude mouse model. Group I (no 9-HpbD-a, no laser irradiation) and Group II (9-HpbD-a injection only) showed no response (4/4, 100%), and Group III (laser irradiation only) showed recurrence (1/4,25%) or no response (3/4, 75 %). Group IV (9-HpbD-a + laser irradiation) showed complete response (10/16, 62.5%), recurrence (4/16, 25%) or no response (2/16, 12.5%). Group IV showed a significant remission rate compared to other groups (p<0.05). These results suggest that 9-HpbD-a is a promising photosensitizer for the future and that further studies on biodistribution, toxicity and mechanism of action would be needed to use 9-HpbD-a as a photosensitizer in the clinical setting.

Antihepatotoxic effect of ethanol extracts from steam-dried ginseng berry on ᴅ-galactosamine/lipopolysaccharide-sensitized mice (ᴅ-galactosamine/lipopolysaccharide로 감작된 급성간독성 마우스 모델에서 인삼열매추출물의 간독성 개선 효과)

  • Jang, Su Kil;Park, Jun Sub;Ahn, Jeong Won;Jo, Boram;Kim, Hyun Soo;Kim, JeongHoon;Kim, Sang Yun;Park, Jung Youl;Lee, Do Ik;Park, Hee Yong;Joo, Seong Soo
    • Korean Journal of Food Science and Technology
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    • v.49 no.6
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    • pp.676-684
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    • 2017
  • The present study aimed to examine the hepatoprotective effects of ethanol extracts from steam-dried ginseng berry (SGBE) in both $\text\tiny{D}$-Galactosamine/Lipopolysaccharide ($\text\tiny{D}$-GalN/LPS)-sensitized mice and in vitro models. Our results clearly demonstrated that SGBE significantly reduced the level of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and lactate dehydrogenase in blood, and $TNF{\alpha}$ was normalized in 8 h after the treatment with $\text\tiny{D}$-GalN/LPS. Coincidently, major organs remained unimpaired when compared to $\text\tiny{D}$-GalN/LPS control group. Moreover, p38, which stimulates expression of NAFLD-associated cytokines, was markedly inhibited when treated with SGBE. In vitro analysis revealed that the main components of SGBE, linoleic acid and ginsenoside Re/Rd, may play a role in protecting liver from $\text\tiny{D}$-GalN/LPS-induced toxicity. Finally, we concluded that SGBE may be a promising therapeutic agent for preventing damage to the liver.

Cell Death Study in Embryonic Stem Cell-derived Neurons and Its Applications (배아줄기세포 유래 신경계세포에서의 세포사멸 연구와 그 응용)

  • Lee, Chul-Sang
    • Development and Reproduction
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    • v.12 no.1
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    • pp.1-8
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    • 2008
  • Specific protocols to increase the differentiation of neuronal cells from embryonic stem (ES) cells have been well established, such as retinoic acid induction and lineage selection of neuronal cells. For the neuropathological studies, ES-derived neurons (ES neurons) must show normal physiological characteristics related to cell death and survival and should be maintained in vitro for a sufficient time to show insults-specific cell death without spontaneous death. When mouse ES cells were plated onto astrocytes monolayer after retinoic acid induction, most ES cells differentiated into neuronal cells, which were confirmed by the presence of specific neuronal markers, and the cultures were viable for at least four weeks. When these cultures were examined for vulnerability to glutamate excitotoxicity, ES neurons were vulnerable to excitotoxic insults mediated by agonist-specific receptors. The vulnerability to excitotoxic death increased with developmental age of ES neurons in vitro. Specific receptors for Neurotrophin and GDNF family ligands were present in ES neurons. GDNF and NT-3 could modulate the survival and excitotoxic vulnerability of ES neurons. The vulnerability and resistance to toxic insults, which are essential requirements of model culture systems for neuropathological studies, make ES neurons to a useful model culture system. Especially ES cell are highly amenable to genetic modification unlikely to primary neuronal cells, which will give us a chance to answer more complicated neurophysiological questions. Recently there was an outstanding attempt to explore the cellular toxicity using human ES cells (Schrattenholz & Klemm, 2007) and it suggested that ES cells could be a new model system for neurophysiological studies soon and go further a large-scale screening system for pharmacological compounds in the future.

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