• Title/Summary/Keyword: environmental DNA

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Isolation of an Isocarbophos-Degrading Strain of Arthrobacter sp. scl-2 and Identification of the Degradation Pathway

  • Li, Rong;Guo, Xinqiang;Chen, Kai;Zhu, Jianchun;Li, Shunpeng;Jiang, Jiandong
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1439-1446
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    • 2009
  • Isocarbophos is a widely used organophosphorus insecticide that has caused environmental pollution in many areas. However, degradation of isocarbophos by pure cultures has not been extensively studied, and the degradation pathway has not been determined. In this paper, a highly effective isocarbophos-degrading strain, scl-2, was isolated from isocarbophos-polluted soil. The strain scl-2 was preliminarily identified as Arthrobacter sp. based on its morphological, physiological, and biochemical properties, as well as 16S rDNA analysis. The strain scl-2 could utilize isocarbophos as its sole source of carbon and phosphorus for growth. One hundred mg/l isocarbophos could be degraded to a non detectable level in 18 h by scl-2 in cell culture, and isofenphos-methyl, profenofos, and phosmet could also be degraded. During the degradation of isocarbophos, the metabolites isopropyl salicylate, salicylate, and gentisate were detected and identified based on MS/MS analysis and their retention times in HPLC. Transformation of gentisate to pyruvate and fumarate via maleylpyruvate and fumarylpyruvate was detected by assaying for the activities of gentisate 1,2-dioxygenase (GDO) and maleylpyruvate isomerase. Therefore, we have identified the degradation pathway of isocarbophos in Arthrobacter sp. scl-2 for the first time. This study highlights an important potential use of the strain scl-2 for the cleanup of environmental contamination by isocarbophos and presents a mechanism of isocarbophos metabolism.

Regulation of CYP 1A1 gene expression by retinoic acid receptor, retinoid X receptor and constitutive androstane receptor in rainbow trout hepatoma cells(RTH 149)

  • Kim, Ji-Sun;Yang, So-Yeun;Seo, Mi-Jung;Sheen, Yhun-Yhong
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.179-179
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    • 2003
  • Exposure of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) causes a variety of biological and toxicology effects, most of which are mediated by aryl hydrocarbon receptor (AhR). The ligand-bound AhR as a heterodimer with AhR nuclear translocator (ARNT) binds to its specific DNA recognition site, the dioxin-responsive element (DRE), and it results in increased transcription of CYP1A1 gene. Retinoic acid (RA) regulates the transcription of various genes for several essential functions through binding to two classes of nuclear receptors, the retinoic acid receptor (RAR) and retinoid X receptor (RXR). Constitutive androstane receptor (CAR) also regulates the transcription of gene. In this study, we have examined how RAR, RXR and CAR regulated CYP1A1 in rainbow trout hepatoma cell (RTH 149) using luciferase reporter gene assay system. We did transient transfection with CYP1A1 luciferase reporter gene and treated with TCDD, all-trans RA, 9-cis RA and phenobarbital. Treatment of all-trans RA, 9-cis RA or phenobarbital decreased the TCDD induced transcription of CYP1A1. When we did transient cotransfection with CYP1A1 luciferase reporter gene and RXR, as increase of RXR concentration, the TCDD induced transcription of CYP1A1 was decreased. Transfection with CAR also decreased the TCDD induced transcription of CYP1A1 in RTH 149 cells.

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New HDAC inhibitor, IN2001 induces apoptosis/cell cycle arrest in human breast cancer cells

  • Euno, Joung-Ki;Nan, Min-Kyung;Jung, Cho-Min;Young, An-Jin;Kim, -Dae-Ki;Yhong, Sheen-Yhun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.180-180
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    • 2003
  • The acetylation of histone is one of the mechanisms involved in the regulation of gene expression and is tightly controlled by two core enzymes, histone acetyltransferase (HAT) and deacetylase (HDAC). There are several reports that imbalance of HAT and HDAC activity is associated with abnormal behavior of the cells in morphology, cell cycle, differentiation, and carcinogenesis. Recently, an increasing number of structurally diverse HDAC inhibitors have been identified that inhibit proliferation and induce differentiation and/or apoptosis of tumor cells in vivo and in vitro. In this study, we have investigated the effects of novel HDAC inhibitors, IN2001 on ER positive and ER negative human breast cancer cell lines. The growth inhibition, cell cycle arrest and apoptosis of cells by HDAC inhibitors were determined using SRB assay, DNA fragmentation, and flow cytometry. We found that IN 2001 as well as Trichostatin A inhibited cell growth dose-dependently in both ER Positive and ER negative human breast cancer cell lines. The growth inhibition with HDAC inhibitors was associated with profound morphological change. The result of cell cycle analysis after 24 h exposure of IN2001 showed G2-M cell cycle arrest in MCF-7 cell and apoptosis in T47B and MDA-MB-231 cell. In summary, IN2001 has antiproliferative effect on human breast cancer cells regardless of the expression of estrogen receptor. These findings heights the possibility of developing HDAC inhibitors as potential anticancer therapeutic agents for the treatment of breast cancer.

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Molecular Systematic Study of Bacterial Community Associated with Sand Dune Plants (사구식물 연관 세균 군집의 분자계통학적 연구)

  • Do, Jin-Ok;Park, Seong-Joo;Kim, Seung-Bum
    • Korean Journal of Environmental Biology
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    • v.25 no.4
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    • pp.356-362
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    • 2007
  • The rhizobacterial diversity associated with 9 native plant species inhabiting coastal sand dunes in Tae-an area, Chungnam Province, was studied using the denaturing gradient gel electrophoresis (DGGE) fingerprinting analysis over three times from October 2003 to March 2004. One dominant band commonly occurred in all of the rhizosphere samples, which was identified as that of Lysobacter enzymogenes. The other common bands included those derived from species of Pseudomonas and Bacillus. It was notable that L. enzymogenes was dominant in all of the 9 plant species and such dominance was consistent throughout the whole sampling period, which confirms the previous study by Lee et al. (2006a). The Bacillus bands were detected in all of the three samplings, and those of Pseudomonas were notable in the samples of December 2003. By the DGGE analysis alone, the significance of Lysobacter to the sand dune plants is not clear. However, considering their presence in healthy plants and the dominance in all plant species, Lysobacter may have positive roles in the survival or growth of the plants in sand dune area.

Molecular Phylogeny of Phytoplakton Isolated from Red Tides in Southern Coast of Korea (국내 남해안에 발생한 적조원인생물들의 24S rRNA 유전자 염기서열분석)

  • Lee, Soo-Woong;Lee, Hee-Woo;Park, Jong-Gyu;Lee, Jin-Ae;Park, Young-Shik
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.3 no.2
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    • pp.90-93
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    • 1998
  • Cultured isolates of Prorocentrum minimum, P. micans, P. triestinum, P. balticum, Gymnodinium sanguineum, Alexandrium catenella, Scrippsiella trochoidea, and Heterosigma akashiwo from red tides in southern coast of Korea were phylogenetically compared on the basis of their 24S rRNA genes. For each isolate approximately 700 bp of 24S rDNA, encompassing D1 and D2 hypervariable domains, was amplified using the polymerase chain reaction and sequenced. The sequences were aligned with those reported in Genbank by using ClustalW program and phylogenetic tree was generated to show that morphospecies designations in the Alexandrium and Prorocentrum species are congruent with terminal taxa defined by phylogenetic analysis of the 24S rRNA fragment. Our results suggest that the molecular phylogeny approach could facilitate identification of domestic dinoflagellates which have ambiguous morphologies.

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The impact of short-term acute heat stress on the rumen microbiome of Hanwoo steers

  • Baek, Youl Chang;Choi, Hyuck;Jeong, Jinyoung;Lee, Sung Dae;Kim, Min Ji;Lee, Seul;Ji, Sang Yun;Kim, Minseok
    • Journal of Animal Science and Technology
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    • v.62 no.2
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    • pp.208-217
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    • 2020
  • Heat stress negatively affects cattle productivity by reducing feed intake. In the present study, we assessed if the rumen microbiome composition of Hanwoo steers was altered by exposure to heat stress. Rumen samples were collected from four Hanwoo steers that were individually housed in climate-controlled chambers with 60% humidity and environmental temperatures of: 1) 15℃ (0-day group), 2) 35℃ for 3 days (3-day group), and 3) 35℃ for 6 days (6-day group). The total community DNA of samples was extracted, and 997,843 bacterial and 1,508,770 archaeal sequences were analyzed using next-generation sequencing. Assessment of the relative abundances revealed 15 major phyla of which Bacteroidetes was found to be the most dominant. After 3 days of heat stress exposure there were no significant changes in the rumen microbiome composition, except for a decrease in the Planctomycetes. However, after 6 days of heat stress exposure, we found that the relative abundance of fibrolytic Ruminococcaceae had decreased while that of lactate-producing Lactobacillaceae and amylolytic Prevotella and Ruminobacter had increased. The normal rumen microbiome of Hanwoo cattle was shown to be disrupted after 6 days of heat stress, which led to the decrease in fibrolytic bacteria that are sensitive to low pH and the increase in both lactate-producing and amylolytic bacteria. We have demonstrated that the microbiome composition of the rumen is affected by acute heat stress. Our findings may contribute to the development of different feeding strategies to restore heat stress-induced disruption of the rumen microbiome.

Cadmium-induced COX-2 Expression in Cerebrovascular Endothelial Cells (카드뮴이 뇌혈관 내피세포에서의 $PGE^2$ 및 COX-2 발현에 미치는 영향)

  • Park Dong-Hyun;Kim Young-Chae;Moon Chang-Kiu;Jung Yi-Sook;Baik Eun-Joo;Moon Chang-Hyun;Lee Soo-Hwan
    • Environmental Analysis Health and Toxicology
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    • v.21 no.3 s.54
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    • pp.275-282
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    • 2006
  • In order to get insight into the mechanism of cadmium (Cd)-induced brain injury, we investigated the effects of Cd on the induction of COX-2 in bEnd.3 mouse brain endothelial cells. Cd induced COX-2 expression and $PGE_2$ release, which were attenuated by thiol-reducing antioxidant N-acetylcysteine (NAC) indicating oxidative components might contribute to these events. Indeed, Cd increased cellular reactive oxygen species (ROS) level and DNA binding activity of nuclear factor-kB (NF-kB), an oxidative stress sensitive transcription factor. Cd-induced $PGE_2$ production and COX-2 expression were significantly attenuated by Bay 11 7082, a specific inhibitor of NF-kB and by SB203580, a specific inhibitor of p38 mitogen activated protein kinase (MAPK). These data suggest that Cd induces COX-2 expression through activation of NF-kB and p38 MAPK, the oxidative stress-sensitive signaling molecules, in brain endothelial cells.

Expression of Rat Hepatic Glutathione-S-Transferases by Benzoazoles (Benzoazole계 화합물이 glutathione-S-transferases의 유도발현에 미치는 영향)

  • 서경원;김연정;김태완;김효정;조민경;김상건
    • Environmental Analysis Health and Toxicology
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    • v.13 no.3_4
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    • pp.55-61
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    • 1998
  • Glutathione-S-transferases (GSTs) detoxify electrophilic xenobiotics and reactive metabolites. Recently benzene-fused heterocycles have been shown to increase the total amount of hepatic GSTs in rats. Primarily this study aimed to determine the induction of GSTs by benzoazoles (BAs) including benzoxazole (BX), 2-methylbenzoxazole (M-BX), 2,5-dimethyl benzoxazole (D-BX), benzothiazole (BT), aminobenzothiazole (A-BT) and 2-mercaptobenzothiazole (M-BT) in rats. Hepatic cytosol and poly(A)$^+$ mRNA were prepared from rats after oral administration of BX, BT, M-BX, D-BX, A-BT and M-BT for 5 consecutive days at doses of 1 mmol/kg. Western immunoblot and northern blot analysis were conducted with rabbit anti-GST Ya, Yb$_1$, Yb$_2$, Yc antibodies and cDNA probes containing = 500 bps in the specific coding regions of Ya, Yb$_1$, Yb$_2$, Yc$_1$, and Yc$_2$, respectively. All BAs increased the amount of enzymes and mRNA levels of GSTs. BT was the most effective inducer of GSTs among the compounds examined in this study. Although A-BT and M-BT, the derivatives of BT, induced GSTs, these chemicals had lesser effect on induction of GSTs than BT. The derivatives of BX also induced less GSTs than the parent compound and the addition of methyl group to the benzene ring of BX reduced the induction of GSTs. BAs had better inductive effects on the class $\alpha$(Ya, Yc) than class $\mu$ GSTs (Yb$_1$, Yb$_2$). BAs enhanced mRNA levels of GSTs in parallel with the protein levels. These results indicate that 1) most of BAs induced various isozymes of GSTs, 2) the induction of GSTs appears to be correlated with the chemical structure of the derivatives, and 3) the expression of GST by BAs is presumably under the transcriptional regulation.

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Anticarcinogenic Activity of a Novel Anthraquinone Derivative DHAQ-97: Induction of Apoptosis in Human Breast Cancer Cell Line MCF-7 (새로운 Anthraquinone 유도체, DHAQ-97의 항암작용: 아폽토시스에 의한 인체 유방암세포 사멸 유도)

  • 허연진;김정환;장정희;안병준;서영준
    • Environmental Mutagens and Carcinogens
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    • v.20 no.1
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    • pp.14-20
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    • 2000
  • DHAQ-97, (2-(3-[p-bis(2-chloroethyl)aminophenyl]-2 formylaminopropanoyloxy) methy1-1,4-dihy-droxy-9,10-anthraquinone), is a novel anthraquinone derivative synthesized for use as an anti-neoplastic agent. In the present study, we have evaluated the selective cytotoxicity of DHAQ-97 by comparing its effects on viability and proliferation of human breast cancer cell line (NCF-7) versus normal immortalized breast epithelial cell line (MCF-10A). Thus, DHAQ-97 reduced both viability and proliferation of MCF-7 cells to a much greater extent than did for MCF-10A cells. The growth inhibitory and anti-proliferative properties of DHAQ-97 appear to be attributable to its ability to induce apoptosis as revealed by positive staining after in 냐셔 nick-end labeling (TUNEL), cleavage of poly(ADP-ribose)polymerase, release of mitochondrial cytochrome c into cytoplasm, and increased expression of pro-apoptotic Bax protein. Recent studies have indicated possible involvement of the ubiquitous eukaryotic transcription factor, NF-kappa B (NF-kB) in the regulation of apoptotic cell death. In line induced cytotoxicity in cultured MCF-7 cells. Furthermore, mild activation of NF-kB, as determined by its increased DNA binding capability, was observed 30 min after treatment with 10$\mu\textrm{m}$ DHAQ-97. Taken together, the above findings suggest that DHAQ-97 exerts selective cytotoxicity towards cancer cells through induction of apoptosis, which appears to be regulated by NF-kB.

Development of distinction methods for male-sterile and dwarfism herbicide tolerant Zoysia japonica Steud (웅성불임 및 왜성형질의 제초제저항성 들잔디(zoysia japonica Steud.)의 판별기술 개발)

  • Lee, Bum Kyu;Kang, Hong-Gyu;Ra, Nu Ri;Sun, Hyeon-Jin;Kwon, Yong-Ik;Song, In-Ja;Kim, Chang-Gi;Ryu, Tae-Hun;Park, Kee Woong;Lee, Hyo-Yeon
    • Korean Journal of Agricultural Science
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    • v.41 no.3
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    • pp.187-191
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    • 2014
  • The cultivation area and use of genetically modified (GM) crops have been increased continuously over the world. Concerns about the potential risks of GM crops are also increasing. Safe management for the development and production of GM crops is required according to Living Modified Organism Act in Korea. Planning about the methods, duration, and frequency of environmental monitoring is also required for commercial use of GM crops. GM Zoysia japonica Steud. (event name: JG21) expressing resistance to glufosinate-ammonium has been generated previously. By using gamma ray treatment to JG21 we also developed male sterility and dwarf Z. japonica (event name: JG21-MS). The objective of this study was to establish the monitoring system for environment release of JG21-MS. In this study we extracted RNA from JG21 and JG21-MS and conducted RAPD (random amplified polymorphic DNA) method to distinguish JG21 and JG21-MS.