• Title/Summary/Keyword: environmental DNA

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Gene Expression Profiling of Eukaryotic Microalga, Haematococcus pluvialis

  • EOM HYUNSUK;PARK SEUNGHYE;LEE CHOUL-GYUN;JIN EONSEON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1060-1066
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    • 2005
  • Under environmental stress, such as strong irradiance or nitrogen deficiency, unicellular green algae of the genus Haematococcus accumulate secondary carotenoids, i.e. astaxanthin, in the cytosol. The induction and regulation of astaxanthin biosynthesis in microalgae has recently received considerable attention owing to the increasing use of secondary carotenoids as a source of pigmentation for fish aquacultures, and as a potential drug in cancer prevention as a free-radical quencher. Accordingly, this study generated expressed sequence tags (ESTs) from a library constructed from astaxanthin-induced Haematococcus pluvialis. Partial sequences were obtained from the 5' ends of 1,858 individual cDNAs, and then grouped into 1,025 non-overlapping sequences, among which 708 sequences were singletons, while the remainder fell into 317 clusters. Approximately $63\%$ of the EST sequences showed similarity to previously described sequences in public databases. H. pluvialis was found to consist of a relatively high percentage of genes involved in genetic information processing ($15\%$) and metabolism ($11\%$), whereas a relatively low percentage of sequences was involved in the signal transduction ($3\%$), structure ($2\%$), and environmental information process ($3\%$). In addition, a relatively large fraction of H. pluvialis sequences was classified as genes involved in photosynthesis ($9\%$) and cellular process ($9\%$). Based on this EST analysis, the full-length cDNA sequence for superoxide dismutase (SOD) of H. pluvialis was cloned, and the expression of this gene was investigated. The abundance of SOD changed substantially in response to different culture conditions, indicating the possible regulation of this gene in H. pluvialis.

Comparative evaluation of the mutagenicity and genotoxicity of smoke condensate derived from Korean cigarettes

  • Kim, Ha Ryong;Lee, Jeong Eun;Jeong, Mi Ho;Choi, Seong Jin;Lee, Kyuhong;Chung, Kyu Hyuck
    • Environmental Analysis Health and Toxicology
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    • v.30
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    • pp.14.1-14.7
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    • 2015
  • Objectives Cigarette smoking is associated with carcinogenesis owing to the mutagenic and genotoxic effects of cigarette smoke. The aim of this study was to evaluate the mutagenic and genotoxic effects of Korean cigarettes using in vitro assays. Methods We selected 2 types of cigarettes (TL and TW) as benchmark Korean cigarettes for this study, because they represent the greatest level of nicotine and tar contents among Korean cigarettes. Mutagenic potency was expressed as the number of revertants per ${\mu}g$ of cigarette smoke condensate (CSC) total particulate matter whereas genotoxic potency was expressed as a concentration-dependent induction factor. The CSC was prepared by the International Organization for Standardization 3308 smoking method. CHO-K1 cells were used in vitro micronucleus (MNvit) and comet assays. Two strains of Salmonella typhimurium (Salmonella enterica subsp. enterica ; TA98 and TA1537) were employed in Ames tests. Results All CSCs showed mutagenicity in the TA98 and TA1537 strains. In addition, DNA damage and micronuclei formation were observed in the comet and MNvit assays owing to CSC exposure. The CSC from the 3R4F Kentucky reference (3R4F) cigarette produced the most severe mutagenic and genotoxic potencies, followed by the CSC from the TL cigarette, whereas the CSC from the TW cigarette produced the least severe mutagenic and genotoxic potencies. Conclusions The results of this study suggest that the mutagenic and genotoxic potencies of the TL and TW cigarettes were weaker than those of the 3R4F cigarette. Further study on standardized concepts of toxic equivalents for cigarettes needs to be conducted for more extensive use of in vitro tests.

Appropriate In Vitro Methods for Genotoxicity Testing of Silver Nanoparticles

  • Kim, Ha Ryong;Park, Yong Joo;Shin, Da Young;Oh, Seung Min;Chung, Kyu Hyuck
    • Environmental Analysis Health and Toxicology
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    • v.28
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    • pp.3.1-3.8
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    • 2013
  • Objectives We investigated the genotoxic effects of 40-59 nm silver nanoparticles (Ag-NPs) by bacterial reverse mutation assay (Ames test), in vitro comet assay and micronucleus (MN) assay. In particular, we directly compared the effect of cytochalasin B (cytoB) and rat liver homogenate (S9 mix) in the formation of MN by Ag-NPs. Methods Before testing, we confirmed that Ag-NPs were completely dispersed in the experimental medium by sonication (three times in 1 minute) and filtration ($0.2{\mu}m$ pore size filter), and then we measured their size in a zeta potential analyzer. After that the genotoxicity were measured and especially, S9 mix and with and without cytoB were compared one another in MN assay. Results Ames test using Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains revealed that Ag-NPs with or without S9 mix did not display a mutagenic effect. The genotoxicity of Ag-NPs was also evaluated in a mammalian cell system using Chinese hamster ovary cells. The results revealed that Ag-NPs stimulated DNA breakage and MN formation with or without S9 mix in a dose-dependent manner (from $0.01{\mu}g/mL$ to $10{\mu}g/mL$). In particular, MN induction was affected by cytoB. Conclusions All of our findings, with the exception of the Ames test results, indicate that Ag-NPs show genotoxic effects in mammalian cell system. In addition, present study suggests the potential error due to use of cytoB in genotoxic test of nanoparticles.

Chestnut Ink Disease Caused by Phytophthora katsurae (Phytophthora katsurae 에 의한 밤나무 잉크병)

  • Oh, E.;Lee, J.-K.;Lee, S.-H.;Kim, K.-H.
    • Journal of Forest and Environmental Science
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    • v.23 no.1
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    • pp.65-71
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    • 2007
  • In early 2000's, about six Phytophthora species have been newly described leading mortality on coniferous and broad-leaved trees in forests. Also, some species of Phytophthora are responsible for ink disease in chestnut plantation near or within forests. Similar symptoms of ink disease were appeared in some areas of Kyungnam and Jeonnam providences in 2005, and the pathogen was isolated using Phytophthora- selective medium in 2006. Morphological and genetic analysis were performed to identify the isolate. Also, the pathogenicity was conducted to complete $K\ddot{o}ch^{\prime}s$ postulate and compare susceptibility among chestnut cultivars. The molecular analysis between P. katsurae and P. hevae were performed with the isolates obtained from different countries including Korea or the sequences downloaded from Phytophthora webpage. The result showed that the isolated pathogen from chestnut was P. katsurae. There is no report of P. katsurae in Korea until now. P. katsurae was re-isolated from inoculated chestnut cultivars. Also, there was a slight difference in susceptibility among chestnut cultivars. The rDNA sequence of our isolate showed 100% similarity with sequence of the isolate cultured from Japan and New Zealand.

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Isolation and Characterization of a Feather-Degrading Bacterium for Recycling of Keratinous Protein Waste (케라틴 단백질 폐기물의 재활용을 위한 우모부해 세균의 분리와 특성)

  • Kim, Jung-Chul;Kim, Min-Ju;Son, Hyeng-Sik;Ryu, Eun-Youn;Jeong, Seong-Yun;Kim, Mi-A;Park, Geun-Tae;Son, Hong-Joo;Lee, Sang-Joon
    • Journal of Environmental Science International
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    • v.16 no.12
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    • pp.1337-1343
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    • 2007
  • The aim of this study was to isolate chicken feather-degrading bacteria with high keratinolytic activity and to investigate cultural conditions affecting keratinolytic enzyme production by a selected isolate. A chicken feather-degrading bacterial strain CH3 was isolated from poultry wastes. Isolate CH3 degraded whole chicken feather completely within 3 days. On the basis of phenotypical and 16S rDNA studies, isolate CH3 was identified as Bacillus thuringiensis CH3. This strain is the first B. thuringiensis described as a feather degrader. The bacterium grew with an optimum at pH 8.0 and $37^{\circ}C$, where maximum keratinolytic activity was also observed. The composition of optimal medium for keratinolytic enzyme production was feather 0.1%, sucrose 0.7%, casein 0.3%, $K_2HPO_4$ 0.03%, $KH_2PO_4$ 0.04%, $MgCl_2$ 0.01% and NaCl 0.05%, respectively. The keratinolytic enzyme had a pH and temperature optima 9.0 and $45^{\circ}C$, respectively. The keratinolytic activity was inhibited ethylenediaminetetraacetic acid, phenylmethylsulfonyl fluoride, and metal ions like $Hg^{2+},\;Cu^{2+}\;and\;Zn^{2+}$. The enzyme activated by $Fe^{2+}$, dithiothreitol and 2-mercaptoethanol.

Anticytotoxic and Radical Scavenging Activities of Acer tegmentosum Maxim Stem Extracts (산겨릅나무 줄기 추출물의 세포독성 억제 및 라디칼 소거 활성)

  • Rhim, Tae-Jin
    • Journal of Environmental Science International
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    • v.24 no.11
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    • pp.1315-1329
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    • 2015
  • The objective of this study was to investigate anticytotoxic and antioxidatative capacities of ethanol extracts from Acer tegmentosum Maxim (A. tegmentosum) stem in vitro. The extract at concentration of 200 ug/mL inhibited 10 and 20 ug/mL arsenic trioxide-induced cytotoxicity of HepG2 cells by 79.3 and 57.5%, respectively. The extract at concentration of 200 ug/mL inhibited 0.2 and 0.5 mM t-BHP-induced cytotoxicity of HepG2 cells by 66.3 and 35.7%, respectively. Antioxidative effects of the extract were examined via measurement of ABTS, superoxide, and peroxyl radical scavenging activities. ABTS radical scavenging activity of the extract was higher than that of ${\alpha}$-tocopherol. Superoxide scavenging activity of the extract was higher than that of catechin. Oxygen radical absorbance capacity of the extract was higher than that of ascorbic acid. Cupric reducing antioxidant capacity of the extract was higher than that of ${\alpha}$-tocopherol. The extract at concentrations of 100 and $500{\mu}g/mL$ inhibited 10 mM t-BHP-induced lipid peroxidation of HepG2 cells by 38.2 and 80.7%, respectively. The extract prevented supercoiled DNA strand breakage induced by hydroxyl or peroxyl radical. Total phenolic and flavonoid contents of the extract at concentration of $100{\mu}g/mL$ were 71.3 nmol/mL gallic acid and 18.8 nmol/mL catechin equivalents, respectively. Thus, strong cytoprotective and antioxidant effects of A. tegmentosum stem extract seem to be due to, at least in part, the prevention from free radicals-induced oxidation as well as high levels in polyphenolic contents.

Research Perspective of an Extremophilic Bacterium, Deinococcus radiodurans on Bioremediation of Radioactive Wastes (방사성 폐기물의 생물정화를 위한 극한세균 데이노코쿠스 라디오두란스의 연구적 고찰)

  • Jeong, Sun-Wook;Choi, Yong Jun
    • Applied Chemistry for Engineering
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    • v.28 no.2
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    • pp.133-140
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    • 2017
  • Increasing concerns on radioactive wastes have drawn much attention on the development of remediation technologies. Massive amounts of radioactive wastes generated from hospital and nuclear power plants were exposed to our environment. Although physicochemical removal methods were developed, an eco-friendly remediation method has not yet been demonstrated. Recently, an extremophilic bacterium has received much attention due to their extraordinary characteristics. Among them, Deinococcus radiodurans (D. radiodurans) strain was regarded as the best host organism for the removal of radioactive heavy metals and radionuclides, because of their superb characteristics like tolerance against the high level of radioactivity. In this article, we briefly introduced the extraordinary nature of D. radiodurans and also discussed the potential use of D. radiodurans strain for the removal of radioactive wastes.

Mitochondrial DNA Sequence Variations and Genetic Relationships among Korean Thais Species (Muricidae: Gastropoda)

  • Lee, Sang-Hwa;Kim, Tae-Ho;Lee, Jun-Hee;Lee, Jong-Rak;Park, Joong-Ki
    • Animal Systematics, Evolution and Diversity
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    • v.27 no.1
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    • pp.1-17
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    • 2011
  • Thais Roding, 1798, commonly known as rock-shell, is among the most frequently found gastropod genera worldwide on intertidal rocky shores including those of Japan, China, Taiwan and Korea. This group contains important species in many marine environmental studies but species-level taxonomy of the group is quite complicated due to the morphological variations in shell characters. This study examined the genetic variations and relationships among three Korean Thais species based on the partial nucleotide sequences of mitochondrial cox1 gene fragments. Phylogenetic trees from different analytic methods (maximum parsimony, neighbor-joining, and maximum likelihood) showed that T. bronni and T. luteostoma are closely related, indicating the most recent common ancestry. The low sequence divergence found between T. luteostoma and T. bronni, ranging from 1.53% to 3.19%, also corroborates this idea. Further molecular survey using different molecular marker is required to fully understand a detailed picture of the origin for their low level of interspecific sequence divergence. Sequence comparisons among conspecific individuals revealed extensive sequence variations within the three species with maximum values of 2.43% in T. clavigera and 1.37% in both T. bronni and T. luteostoma. In addition, there is an unexpectedly high level of mitochondrial genotypic diversity within each of the three Korean Thais species. The high genetic diversity revealed in Korean Thais species is likely to reflect genetic diversity introduced from potential source populations with diverse geographic origins, such as Taiwan, Hong Kong, and a variety of different coastal regions in South China and Japan. Additional sequence analysis with comprehensive taxon sampling from unstudied potential source populations will be also needed to address the origin and key factors for the high level of genetic diversity discovered within the three Korean Thais species studied.

Analysis of a Microbial Community Denitrying Nitrate to Nitrogen Gas in a Nitrate-Contaminated Aquifer

  • Jin-Hun, Kim;Bong-Ho, Son;Su-Yeol, Gwon;Seong-Uk, Eo;Yeong, Kim
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.09a
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    • pp.175-178
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    • 2004
  • Little study has been published specifically addressing the dynamics of nitrate reducing bacteria (NBR) during the bioremediation of nitrate-contaminated aquifer. In our previous study we successfully quantified fumarate-enhanced microbial nitrate reduction rate in a nitrate-contaminated aquifer by using a series of single-well push-pull tests (PPTs). In this study we analyzed the suspended population during PPTs. To monitor changes in the microbial community, PCR amplification of 16S rDNA genes and denaturing gradient gel electrophoresis (DGGE) were used to study the dynamics of the bacterial community in detail. Before the stimulation of NBR, the dominant DGGE bands obtained by PCR were affiliated with V-Proteobacteria consisting of Acinetobacter spp. and Pseudomonas fluorescens. However, as NBR biostimulation proceeded, the dominant patterns of DGGE bands changed, and they were affiliated with Azoarcus denitrificans Td-3 and Flavobacterium xanthum. Azoarcus denitrificans Td-3 is known to completely reduce nitrate to nitrogen gas. The series of single-well push-pull tests in this study should prove useful for conducting rapid, low-cost feasibility assessments for in situ denitrification and provide important information about which microorganisms play a key role in bioremediation of a nitrate contaminated aquifer.

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The Relationship between Urinary 8-hydroxydeoxyguanosine and Polycyclic Aromatic Hydrocarbons Exposure in Working Environment

  • Lee Jong-Seong;Kim Eun-A;Lee Yong-Hag;Moon Deog-Hwan;Kim Kwang-Jong
    • Biomedical Science Letters
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    • v.11 no.2
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    • pp.143-152
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    • 2005
  • To investigate the exposure effect of polycyclic aromatic hydrocarbons (PAHs), we analyzed the relationship between urinary 8-hydroxydeoxyguanosine (8-OHdG) concentration and PAHs exposure. The study population contained 44 workers in steel-pipe coating and paint manufacture industries. We measured airborne total PAHs as an external dose, urinary 1-hydroxypyrene (1-OHP) as an internal dose of PAHs exposure, and urinary 8-OHdG as an effective dose of oxidative DNA damage. There was significant correlation between the urinary concentration of l-OHP and the environmental concentration of PAHs, pyrene, urinary cotinine, AST, and GGT. The mean of urinary 8-OHdG was $17.07\pm1.706{\mu}g/g$ creatinine in workers exposed to airborne PAHs. There was significant correlation between the urinary concentration of 8-0HdG and the airborne concentration of PAHs. From the results of stepwise multiple regression analysis about 8-OHdG, significant independents was total PAHs. In this study, there were significant correlation between the urinary concentration of 8-OHdG and the airborne concentration of PAHs. The urinary 1-OHP was effective index as a biomarker of airborne PAHs in workplace. But it was influenced by non-occupational PAHs source, smoking and biomarkers of liver function test.

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