• Title/Summary/Keyword: environmental DNA

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[6]-Gingerol Inhibits Phorbol Ester-Induce d Expression of Cyclooxygenase-2 in Mouse Skin: p38 MAPK and p65/RelA as Possible Molecular Targets

  • Kim, Sue-Ok;Chun, Kyung-Soo;Surh, Young-Joon
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2003.05a
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    • pp.95.1-95
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    • 2003
  • Ginger (Zingiber officinale Roscoe, Zingiberaceae) has a wide array of pharmacologic effects. Our previous studies have demonstrated that [6]-gingerol, a major pungent ingredient of ginger, inhibits mouse skin tumor promotion and anchorage-independent growth of cultured mouse epidermal cells stimulated with epidermal growth factor. In this study, we have investigated the molecular mechanisms underlying chemopreventive effects of [6]-gingerol on mouse skin carcinogenesis. Cyclooxygenase-2 (COX-2), a key enzyme in the formation of prostaglandins, has been recognized as a molecular target of many chemopreventive as well as anti-inflammatory agents. The murine COX-2 promoter contains several transcriptional elements, particularly those involved in regulating inflammatory processes. One of the essential transcription factors responsible for COX-2 induction is NF-kappa B. Topical application of [6]-gingerol inhibited the COX-2 expression through suppression of NF-kappa B activation in phorbol ester-treated mouse skin. [6]-Gingerol, through down-regulation of p38 MAPK, abrogated the DNA binding activity of NF-kappa B by blocking phosphorylation of p65/RelA at the Ser 536 residue. These findings suggest that [6]-gingerol exerts an anti-tumor promotional activity through inhibition of the p38 MAPK-NF-kappa B siganling cascade in mouse skin.

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Assessment of genetic diversity and distance of three Cicuta virosa populations in South Korea

  • Nam, Bo Eun;Kim, Jae Geun;Shin, Cha Jeong
    • Journal of Ecology and Environment
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    • v.36 no.3
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    • pp.205-210
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    • 2013
  • Cicuta virosa L. (Apiaceae) is a perennial emergent plant designated as an endangered species in South Korea. According to the former records, only four natural habitats remain in South Korea. A former study suggested that three of four populations (Pyeongchang: PC, Hoengseong: HS, Gunsan: GS) would be classified as different ecotypes based on their different morphological characteristics and life cycle under different environmental conditions. To evaluate this suggestion, we estimated genetic diversity in each population and distance among three populations by random amplification of polymorphic DNA. Seven random primers generated a total of 61 different banding positions, 36 (59%) of them were polymorphic. Nei's gene diversity and the Shannon diversity index increased in the order of PC < HS < GS, which is the same order of population size. In the two-dimensional (2D) plot of first two principal components in principal component analysis with the presence of 61 loci, individuals could be grouped as three populations easily (proportion of variance = 0.6125). Nei's genetic distance for the three populations showed the same tendency with the geographical distance within three populations. And it is also similar to the result of discriminant analysis with the morphological or life-cycle factors from the previous study. From the results, we concluded that three different populations of C. virosa should be classified as ecotypes based on not only morphology and phenology but genetic differences in terms of diversity and distance as well.

Effects of habitat differences on the genetic diversity of Persicaria thunbergii

  • Nam, Bo Eun;Nam, Jong Min;Kim, Jae Geun
    • Journal of Ecology and Environment
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    • v.40 no.2
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    • pp.84-88
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    • 2016
  • To understand the effects of habitat characteristics on the genetic diversity of Persicaria thunbergii, three sites of different environmental conditions in a water system were surveyed. Site A was the closest to the source of the water system, and there was a dam between sites A and B. Site C is located on the lowest downstream in the water system. Vegetation survey of four quadrats at each site was performed, and soil samples were collected for physicochemical analysis. Random amplification of polymorphic DNA (RAPD) analysis of ten P. thunbergii individuals at each site was conducted to calculate population genetic diversity and genetic distance among populations. Soil was sterile sand at site A, whereas loamy soil at sites B and C. A pure stand of P. thunbergii appeared at site A, while other species occurred together (such as Humulus japonicus and Phragmites australis) at sites B (Shannon-Wiener index; $H_B=0.309$) and C ($H_C=0.299$). Similar to the species diversity, genetic diversity (Nei's gene diversity; h) within population of site A ($h_A=0.2381$) was relatively lower than sites B ($h_B=0.2761$) and C ($h_C=0.2618$). However, site C was separated from sites A and B in genetic distance rather than the geographical distance (Nei's genetic distance; A~B, 0.0338; B~C, 0.0685; A~C, 0.0833).

An advanced tool, droplet digital PCR (ddPCR), for absolute quantification of the red-tide dinoflagellate, Cochlodinium polykrikoides Margalef (Dinophyceae)

  • Lee, Hyun-Gwan;Kim, Hye Mi;Min, Juhee;Kim, Keunyong;Park, Myung Gil;Jeong, Hae Jin;Kim, Kwang Young
    • ALGAE
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    • v.32 no.3
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    • pp.189-197
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    • 2017
  • To quantify the abundance of the harmful dinoflagellate Cochlodinium polykrikoides in natural seawaters, we developed the innovative procedure using a droplet digital PCR (ddPCR) with C. polykrikoides-specific primers targeting the internal transcription sequence (ITS). The abundance of C. polykrikoides was estimated by the specific copy number of target ITS DNA segments per cell in cultures and natural water samples. The copy number per C. polykrikoides cell as acquired by ddPCR was $157{\pm}16$, which was evaluated against known cell numbers through a simplified protocol preparing DNAs. The abundances of C. polykrikoides in the waters of different locations estimated by ddPCR agreed with the number of cells visually counted under a microscope. This protocol was used to measure the abundance of C. polykrikoides close to and further off the southern coast of Korea in August of 2016 and 2017. The practical application showed that this method can reduce time for analysis and increase accuracy.

Isolation and Proteomic Analysis of a Chlamydomonas reinhardtii Mutant with Enhanced Lipid Production by the Gamma Irradiation Method

  • Baek, Jaewon;Choi, Jong-il;Park, Hyun;Lim, Sangyong;Park, Si Jae
    • Journal of Microbiology and Biotechnology
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    • v.26 no.12
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    • pp.2066-2075
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    • 2016
  • In this study, an enhanced lipid-producing mutant strain of the microalga Chlamydomonas reinhardtii was developed by gamma irradiation. To induce the mutation, C. reinhardtii was gamma irradiated at a dose of 400 Gy. After irradiation, the surviving cells were stained with Nile red. The mutant (Cr-4013) accumulating 20% more lipid than the wild type was selected. Thin-layer chromatography revealed the triglyceride and free fatty acid contents to be markedly increased in Cr-4013. The major fatty acids identified were palmitic acid, oleic acid, linoleic acid, and linolenic acid. Random amplified polymeric DNA analysis showed partial genetic modifications in Cr-4013. To ascertain the changes of protein expression in the mutant strain, two-dimensional electrophoresis was conducted. These results showed that gamma radiation could be used for the development of efficient microalgal strains for lipid production.

The Effect of Jazf1 Overexpression in Zebrafish Cardiac Development

  • Shin, Mi-Jung;Yuh, Hyung-Soo;Seo, Byoung-Boo;Park, Hum-Dai;Yoon, Du-Hak;Ryoo, Zae-Young
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.457-461
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    • 2011
  • JAZF1 (Juxtaposed with Another Zinc Finger gene 1) transcription factor are Zn-finger proteins that bind to the nuclear orphan receptor TAK/TR4 (Nakajima et al., 2004). The nuclear orphan receptor TAK1/TR4 functions as a positive as well as a negative regulator of transcription. It was recently reported that congenital cardiovascular malformations are significantly more frequent in Neurofibromatosis 1 (NF1) patients with microdeletion syndrome than in those with classical NF1. JAZF1 was expressed in adult heart of patients with microdeletion syndrome. JAZF1 is highly conserved among various species include zebrafish. We hypothesized that the expression of zebrafish Jazf1 may lead to severe forms of congenital heart disease that allow the survival of newborns and adults. In this study, we created Jazf1 transgenic zebrafish which over-express zebrafish Jazf1 cDNA under control of the CMV promoter. Our results suggested that Jazf1 expression may play an important role in zebrafish cardiac development.

C-Ring Cleavage of Isoflavones Daidzein and Genistein by a Newly-Isolated Human Intestinal Bacterium Eubacterium ramulus Julong 601

  • Wang, Xiu-Ling;Kim, Ki-Tae;Lee, Je-Hyeon;Hur, Hor-Gil;Kim, Su-Il
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.766-771
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    • 2004
  • Julong 601, a Gram-positive anaerobic bacterium strain capable of cleaving the C-ring of isoflavones daidzein and genistein, was isolated from human feces. BLAST search revealed that its complete 16S rDNA gene sequence has 99% similarity to Eubacterium ramulus. Metabolites of daidzein and genistein were determined as O-desmethylangolensin (O-Dma) and 2-(4-hydroxyphenyl) propionic acid (2-HPPA), respectively, based on UV, EI-MS, and $^1H$ and ^{13}C$ NMR spectral analyses. Enantiomers of O-Dma and 2-HPPA were isolated by chiral stationary-phase HPLC (CSP HPLC). Cleavage of the C-ring of daidzein and genistein by strain Julong 601 was highly enantioselective. Specific rotation ([$\alpha]_D$) and circular dichroism (CD) spectra of the enantiomers are reported here for the first time. Biotransformation kinetics of daidzein and genistein indicated that the C-ring of genistein has a higher susceptibility to bacterial degradation than that of daidzein.

Inhibition of p65 Nuclear Translocation by Baicalein

  • Seo, Min-Bum;Lee, Seog-Ki;Jeon, Young-Jin;Im, Jin-Su
    • Toxicological Research
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    • v.27 no.2
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    • pp.71-76
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    • 2011
  • We demonstrate that baicalein, a bioactive flavonoid originally isolated from Scutellaria baicalensis, inhibits LPS-induced expression of iNOS gene in RAW 264.7 cells. Treatment of peritoneal macrophages and RAW 264.7 cells with baicalein inhibited LPS-stimulated nitric oxide production in a dose-related manner. Immunohistochemical staining of iNOS and RT-PCR analysis showed that the decrease of NO was due to the inhibition of iNOS gene expression in RAW 264.7 cells. Immunostaining of p65, EMSA, and reporter gene assay showed that baicalein inhibited NF-${\kappa}$B nuclear translocation, DNA binding, and transcriptional activation, respectively. Collectively, these series of experiments indicate that baicalein inhibits iNOS gene expression by blocking NF-${\kappa}$B nuclear translocation. Due to the critical role that NO release plays in mediating inflammatory responses, the inhibitory effects of baicalein on iNOS suggest that baicalein may represent a useful anti-inflammatory agent.

Genotoxicity Assessment of Erythritol by Using Short-term Assay

  • Chung, Young-Shin;Lee, Michael
    • Toxicological Research
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    • v.29 no.4
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    • pp.249-255
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    • 2013
  • Erythritol is a sugar alcohol that is widely used as a natural sugar substitute. Thus, the safety of its usage is very important. In the present study, short-term genotoxicity assays were conducted to evaluate the potential genotoxic effects of erythritol. According to the OECD test guidelines, the maximum test dose was 5,000 ${\mu}g$/plate in bacterial reverse mutation tests, 5,000 ${\mu}g/ml$ in cell-based assays, and 5,000 mg/kg for in vivo testing. An Ames test did not reveal any positive results. No clastogenicity was observed in a chromosomal aberration test with CHL cells or an in vitro micronucleus test with L5178Y $tk^{+/-}$ cells. Erythritol induced a marginal increase of DNA damage at two high doses by 24 hr of exposure in a comet assay using L5178Y $tk^{+/-}$ cells. Additionally, in vivo micronucleus tests clearly demonstrated that oral administration of erythritol did not induce micronuclei formation of the bone marrow cells of male ICR mice. Taken together, our results indicate that erythritol is not mutagenic to bacterial cells and does not cause chromosomal damage in mammalian cells either in vitro or in vivo.

Effect of Orostachys japonicus A. Berger on Apoptosis Induction of Human Leukemia HL60 Cells (와송의 HL60백혈병세포의 Apoptosis유도 효과)

  • Oh, Chan-Ho;Bae, Jin-Beom;Kim, Nam-Seok;Jeon, Hoon;Han, Kwang-Soo;Lee, Moon-Jun;Kwon, Jin
    • Korean Journal of Pharmacognosy
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    • v.40 no.2
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    • pp.118-122
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    • 2009
  • Methanol extracts of Orostachys japonicus A. Berger (OAB) were found to exhibit apoptosis induction of HL60 human acute promyelocytic leukemia cells. Treatment of OAB exerted strong cytotoxicity against HL60 cells. OAB induced DNA fragmentation of HL60 cells in a dose dependent manner. Nitric oxide production were also increased in OAB-treated RAW264.7 macrophage cell lines. Treatment of OAB increased the expression of p53 and iNOS gene and the expression of p53, $NF-{\kappa}B$ and iNOS protein in cultured HL60 and RAW264.7 cells. These results suggest that OAB are effective on strong anti-cancer properties and can be useful as a chemo-preventive agents.