• Title/Summary/Keyword: Assay system

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Genotoxicological Safety of Gamma-Irradiated Kwamegi(semi-dried Colobabis seira) (감마선 조사된 과메기의 유전독성학적 안전성 평가)

  • 육홍선;정영진;송현파;이주운;변명우
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.1
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    • pp.182-192
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    • 2004
  • Gamma irradiation at 5 and 10 kGy was applied to Kwamegi (semi-dried Colobabis seira) for their possible hygiene quality and carried out genotoxicological safety. In vitro genotoxicological safety of each 5 and 10 kGy-irradiated Kwamegi was evaluated by Salmonella typhimurium (TA98, TA100, TA1535 and TA1537) and E. coli WP2 uvrA reversion assay, SOS chromotest (Escherichia coli PQ37) and chromosome aberration test (Chinese hamster lung fibroblast cells) in the absence and presence of an exogenous metabolizing system (S9 mix). Gamma-irradiated samples were not different from nonirradiated-control to respective in vitro tests. And in vivo micronucleus test using ICR mice (male) micronucleus was not observed. Kwamegi exposed to 10 kGy-gamma ray revealed negative results in these three in vitro mutagenetic tests and in vivo micronucleus test up to 10,000 $\mu\textrm{g}$/plate, respectively. The results indicated that 5 and 10 kGy gamma-irradiated Kwamegi (semi-dried Colobabis seira) did not have mutagenicity.

The Neurotoxicological Alterations Induced by Narcotic Drugs and Industrial Chemicals in the Rat are Associated with Quantitative Changes in Glial Fibrillary Acidic Protein (마약류 및 산업환경화학물질에 의한 GFAP의 신경독성표지물질화에 관한 유용성)

  • Cho, Dae-Hyun;Jeong, Yong;Kim, Jun-Gyou;Lee, Bong-Hun;Hwang, Se-Jin;Lee, Won-Yong;Kim, Jeong-Goo;Cho, Tai-Soon;Kim, Jin-Suk;Moon, Hwa-Hwey
    • Toxicological Research
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    • v.11 no.2
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    • pp.315-327
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    • 1995
  • Diverse neurotoxic insults result in proliferation and hypertrophy of astrocytes, a subtype of glia in central nervous system. The hallmark of this response, often terms "reactive gliosis", is the enhanced expression of the major intermediate filament protein of castrocytes, glial fibrillary acidic protein (GFAP). These changes in the astrocytes suggest that GFAP may be a useful biochemical indicator of neurotoxicity. To investigate this possibility, we administered intra-peritoneally prototype nerotoxicants, metharnphetamine (MAP, 5 mg/kg), cocaine (30 mg/kg), N-buthyl benzenesulfonamide (NBBS, 300 mg/kg) and trimethytin (TMT, 8 mg/kg) to Wistar Rats and then assessed the effects of these agents on content of GFAP, which were determined by Sandwish ELISA and evaluated with neurotoxic symptoms, and quantitative changes of imrnunoreactivity of GFAP by light microscopic image analysis in specific regions. We found that assay of GFAP revealed time- and region-dependant patterns of neurotoxicity. The GFAP immunoreactivity of rat brain was increased in substantia nigra and hippocampus by MAP, NBBS and TMT; in roedial septal nucleus and nucleus accurnbens, it was also increased by RrBBS. Sandwich ELISA showed that GFAP levels of cerebrum in all groups on days 3 and 7 and that of brainstem(including cerebellum) in MAP, NBBS groups on day 1 and 3 were increased. A review of the background, design and results of these experiments are presented in this paper. Our findings indicate that GFAP is a sensitive and specific biomarker of neurotoxicity.otoxicity.

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Development of herbicide tolerant soybean using Agrobacterium tumefaciens (아그로박테리움을 이용만 제초제 저항성 콩 개발)

  • Lee, Ki-Jong;Park, Hong-Jae;Yi, Bu-Young;Lee, Kyeong-Ryeol;Kim, Myung-Sik;Woo, Hee-Jong;Jin, Yong-Moon;Kweon, Soon-Jong
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.69-74
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    • 2008
  • This study aims to establish the efficient soybean transformation system and develop soybean [Glycine max (L.) Merill] transformants using cotyledonary node explants. The cotyledonary node of soybean were co-cultivated with Agrobacterium tumefaciens strains (KYRT1, EHA105). These strains contain the binary vector pCAMBIA3301 which carries a herbicide-resistant far gene. Korean cultivars (Danbaekkong, Eunhakong) and foreign cultivars (Jack, Peking) were the most efficient in regenerating cotyledonary node. Therefore, they were chosen for the transformation. Results showed that the T-DNA transfer reached up to 60% and transformation efficiency reached up to 3% in the cotyledonary node explants from Jack cultivar, co-cultivated with EHA105 strain. Histochemical GUS evaluation showed that 12 individual lines, transformed with the 현 gene, have positive response. The transformed soybeans have been confirmed in the $T_0$ generation through phenotypic assay using herbicide $Basta^{(R)}$ and Southern blot analysis.

Anticancer and Antioxidant Effects of Solvent Extracts from Dried Onion with Different Drying Methods (건조방법에 따른 건조 양파 추출물의 항암 및 항산화 효과)

  • Jang, Joo-Ri;Kim, Kyun-Kun;Lim, Sun-Young
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1271-1277
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    • 2008
  • We investigated to determine the inhibitory effects of solvent extracts from dried onion on growth of cancer cell lines (HT-1080 human fibrosarcoma and HT-29 human colon cancer cells) and $H_{2}O_{2}$-induced oxidative stress. Two different drying methods, low temperature vacuum dryer and freeze dryer, were employed to dry onion. Inhibitory effects of acetone with methylene chloride (A+M) and methanol (MeOH) extracts from onion by two drying methods on the growth of HT-1080 and HT-29 cancer cells increased in a dose dependent manner (p<0.05) and the higher inhibitory effect was shown in onion extracts dried by low temperature vacuum dryer. The treatments of hexane, 85% aq. methanol, butanol and water fractions significantly inhibited the growth of both cancer cells (p<0.05) and onion fractions dried by freeze dryer showed a higher inhibitory effect compared with those dried by low temperature vacuum dryer. In order to determine a protective effect on H2O2-induced oxidative stress, DCHF-DA (dichlorodihydrofluorescin diacetate) assay was conducted. All fractions including crude extracts of dried onion appeared to significantly reduce the levels of intracellular reactive oxygen species (ROS) (p<0.05). Higher antioxidant effect was observed in onions dried by the low temperature vacuum dryer method. These results indicate that the low temperature vacuum dryer is useful to dry and produce onion powder.

Involvement of a LiCl-Induced Phosphoprotein in Pigmentation of the Embryonic Zebrafish (Danio rerio) (LiCl에 의해 유도되는 phosphoprotein이 embryonic zebrafish (Danio rerio)의 pigmentation에 미치는 영향)

  • Jin, Eun-Jung;Thibaudeau, Giselle
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1219-1224
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    • 2008
  • The embryonic zebrafish (Danio rerio) is rapidly becoming an important model organism for studies of early events in vertebrate development. Neural crest-derived pigment cell precursors of the embryonic zebrafish give rise to melanophores, xanthophores, and/or iridophores. Cell-signaling mechanisms related to the development of pigmentation and pigment pattern formation remain obscure. In this study, zebrafish embryos were treated with various signaling-related molecules - LiCl (an inositol-phosphatase inhibitor), forskolin (a protein kinase-A activator), a combination of LiCl/forskolin, and LiCl/heparin (an IP3 inhibitor) in order to identify the mechanisms involved in pigmentation. LiCl treatment resulted in ultrastructural and morphological alterations of melanophores. To identify the possible proteins responsible for this ultrastructural and morphological change, phosphorylation patterns in vitro and in vivo were analyzed. LiCl and LiCl/forskolin treatment elicited dramatic increases in the phosphorylation of a 55-kDa protein which was inhibited by heparin treatment. LiCl treatment also induced phosphorylation of a 55-kDa protein in melanophores purified from adult zebrafish. Collectively these results suggest that a LiCl-induced 55-kDa phosphoprotein plays a role in melanophore morphology and ultrastructure and ultimately effects gross pigmentation.

The Significance of SDF-1α-CXCR4 Axis in in vivo Angiogenic Ability of Human Periodontal Ligament Stem Cells

  • Bae, Yoon-Kyung;Kim, Gee-Hye;Lee, Jae Cheoun;Seo, Byoung-Moo;Joo, Kyeung-Min;Lee, Gene;Nam, Hyun
    • Molecules and Cells
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    • v.40 no.6
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    • pp.386-392
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    • 2017
  • Periodontal ligament stem cells (PDLSCs) are multipotent stem cells derived from periodontium and have mesenchymal stem cell (MSC)-like characteristics. Recently, the perivascular region was recognized as the developmental origin of MSCs, which suggests the in vivo angiogenic potential of PDLSCs. In this study, we investigated whether PDLSCs could be a potential source of perivascular cells, which could contribute to in vivo angiogenesis. PDLSCs exhibited typical MSC-like characteristics such as the expression pattern of surface markers (CD29, CD44, CD73, and CD105) and differentiation potentials (osteogenic and adipogenic differentiation). Moreover, PDLSCs expressed perivascular cell markers such as NG2, ${\alpha}-smooth$ muscle actin, platelet-derived growth factor receptor ${\beta}$, and CD146. We conducted an in vivo Matrigel plug assay to confirm the in vivo angiogenic potential of PDLSCs. We could not observe significant vessel-like structures with PDLSCs alone or human umbilical vein endothelial cells (HUVECs) alone at day 7 after injection. However, when PDLSCs and HUVECs were co-injected, there were vessel-like structures containing red blood cells in the lumens, which suggested that anastomosis occurred between newly formed vessels and host circulatory system. To block the $SDF-1{\alpha}$ and CXCR4 axis between PDLSCs and HUVECs, AMD3100, a CXCR4 antagonist, was added into the Matrigel plug. After day 3 and day 7 after injection, there were no significant vessel-like structures. In conclusion, we demonstrated the perivascular characteristics of PDLSCs and their contribution to in vivo angiogenesis, which might imply potential application of PDLSCs into the neovascularization of tissue engineering and vascular diseases.

Cellular Protective Effects and Antioxidative Activity of Resveratrol (레스베라트롤의 세포보호 및 항산화 효과)

  • Jo, Na Rae;Park, Su Ah;Jeon, So Ha;Ha, Ji Hoon;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.24 no.5
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    • pp.483-488
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    • 2013
  • In this study, the cellular protective effect of resveratrol on oxidative damage and its antioxidative activity were investigated. The free radical-scavenging activity ($FSC_{50}$) of resveratrol was measured to be $103{\mu}M$. The reactive oxygen species-scavenging activity ($OSC_{50}$) of resveratrol on the ROS generated in a $Fe^{3+}-EDTA/H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. Resveratrol displayed $0.042{\mu}M$ ROS scavenging activity, which is 9.6-fold higher than that of L-ascorbic acid ($0.405{\mu}M$) and had a more prominent cellular protective effect than (+)-${\alpha}$-tocopherol. When HaCaT cells were exposed to $800mJ/cm^2$ of UVB or treated with $30{\mu}M$ rose bengal, resveratrol protected the cells against oxidative stress in a concentration-dependent manner; however, it was unable to protect the cells when the damage was induced by 10 mM $H_2O_2$. These results indicate that resveratrol could be employed to improve and prevent the skin aging through its antioxidative and cellular protective activities.

Effects of Helicobacter pylori Antigen on Producton of Transforming growth factor-$\beta$1 and Nitric oxide in Human Fibroblast (사람성유아세포의 Transforming growth factor-$\beta$1과 Nitric oxide 생성에 미치는 Helicobacter pylori 항원의 효과)

  • 박무인;박선자;구자영;김광혁
    • Journal of Life Science
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    • v.11 no.2
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    • pp.181-189
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    • 2001
  • Cytokines are hormone-like proteins which mediate and regulast inflammatory and immune responses. Transforming growth factor -$\beta$1(TGF-$\beta$) plays an important role in the control of the immune response and wound healing, and in the development o various tissues and organs, Nitric oxide(NO) is major messenger molecule regulating immune function and blood vessel dilation and serving as a neurotransmitter in the brain and peripheral nervous system. Also, NO is to be a potent mutagen that cause mutation in the p53 tumor suppressor gene in early phases of human gastric carcinogenesis. The purpose of this study was to investigate the effect of Helicobacter phlori lystes, lipopolysaccharide (LPS), and Staphylococcus enterotoxin B(SEB) on production of TGF-$\beta$1 and NO by human fibroblasts. Primary cultured human fibroblasts were incubated with H. pylori lysates(Hp), LPs, SEB, Hp+LPS, Hp+SEB, Hp+LPS+SEB. Cultured supernatants that were collected at 24, 48 and 72 hr were assessed for TGF-$\beta$1 by enzyme-linked immunosorbent assay and NO production by quantification of nitrite ion. TGF-$\beta$1 production in fibroblasts exposed with Hp, LPS or SEB for 48 hrs was enhanced, but for 72 hrs inhibited. Its production by doble exposure such as Hp+LPS, Hp+SEB, Hp+LPS+SEB was lowered in comparison with single exposure of Hp in cases of 24 and 48 hrs incubation, but for 72 hrs decreased in Hp vaculoating toxin(+), increased in Hp vacuolating toxin(-). No production in fibroblasts increaed at all doses of LPS. But its production by exposure of SEB increased or decreased according to dose and incubation time. Also, NO production by Hp vacuolating toxin(+) increased at all doses, but its production by Hp vacuolating toxin(-) decreased. Its production by doble exposure such as Hp+LPS, Hp+SEB, Hp+LPS+SEB decreased in comparison with single exposure Hp Therefore, quantities pf TGB-$\beta$1 and NO released by human fibroblasts shows differences according to kinds of stimulants. Also, in care stimulated with same kinds of stimulants, its productions exhibit quantitative differences according to exposure times. These results suggest that the decreased of TGF-$\beta$1 in fibroblasts by mixed exposure with Hp producing vacuolating toxin and bacterial toxins such as LPS and SEB may effect negatively in healing of host tissue and increased of NO by infection oh H. pylori may related to the increased susceptibility for human gastric carcinogenesis.

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Studies on Antioxidant Activity and Inhibition of Nitric Oxide Synthesis of Germinated Brown Rice Soaked in Mycelial Culture Broth of Phellinus linteus (상황버섯균사체배양액에 침지한 발아현미의 항산화 및 nitric oxide 합성저해에 관한 연구)

  • Jung, Il-Sun;Kim, Yu-Jung;Choi, In-Soon;Choi, Eun-Young;Shin, Su-Hwa;Gal, Sang-Wan;Choi, Young-Ju
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1141-1146
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    • 2007
  • This study investigated the effects on the biological activities of germinated brown rice soaked in mycelial culture broth of Phellinus linteus. The level of free amino acid was higher in the GBRP extract than those of BR and GBR. The major free amino acids were alanine, valine, isoleucine and methionine in both extracts. The level of ${\gamma}$-aminobutyric acid (GABA) was also increased significantly in the GBR and GBRP. Antioxidant activities of methanol extract of BR, GBR and GBRP were measured by using DPPH radical scavenging and SOD-like activity. Antioxidant activities showed the highest level of 83% and 76% when 100 mg/ml GBR and GBRP, respectively. Stimulation of the macrophages RAW264.7 cells with lipopolysaccharide (LPS) resulted in increased production of nitric oxide (NO) in the medium. However, the methanol extract of GBR and GBRP showed marked inhibition of NO synthesis in a does-dependant manner. These results showed that GBR and GBRP were significant role for activation of immune system in the pathogenesis of inflammatory diseases.

Antibacterial Effects of Pseudomonas aeruginosa MB I-3 against Listonella anguillarum (어류 병원세균, Listonella anguillarum에 대한 Pseudomonas aeruginosa MB I-3의 항균 효과)

  • Lee, Su-Jung;Youn, I Na;Kim, Jin-Do;Lee, Jung Sick;Kim, Eunheui
    • Journal of fish pathology
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    • v.27 no.1
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    • pp.17-24
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    • 2014
  • To study the possible use of probiotics in fish farming, The in vitro and in vivo antibacterial effects of Pseudomonas aeruginosa MB I-3 (MB I-3) against the fish pathogenic bacterium Listonella anguillarum were evaluated. The inhibitory effects of MB I-3 against vibrios were investigated by the double layer method and the co-culture. The results showed that MB I-3 inhibited the growth of pathogenic vibrios including Listonella anguillarum, Vibrio alginolyticus, Vibrio cholerae, Vibrio fluvialis, Vibrio furnissii, Vibrio harveyi, Vibrio parahaemolyticus and Vibrio vulnificus. Extracellular substances obtained from the cultural supernatant of MB I-3 by ethyl acetate extraction showed inhibitory effects on L. anguillarum. The antibacterial substance of MB I-3 was evaluated to destroy the cell membrane of L. anguillarum in electron micrographs. The probiotic effects of MB I-3 was tested by exposing olive flounder (Paralichthys olivaceus) fry to L. anguillarum with or without MB I-3. The cumulative mortality of olive flounder fry infected with L. anguillarum was 24% in the group with MB I-3, while it was 46% in the control group without MB I-3. These results indicate that MB I-3 has potential applications as a probiotic for the control of fish pathogenic vibrios in fish rearing system.