• Title/Summary/Keyword: environmental DNA

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Prediction of Photo-Carcinogenicity from Photo-Ames Assay (Photo-Ames Assay를 이용한 광발암성 예측)

  • Hong Mi Young;Kim Ji Young;Chung Moon Koo;Lee Michael
    • Environmental Mutagens and Carcinogens
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    • v.25 no.1
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    • pp.6-12
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    • 2005
  • Many compounds might become activated after absorption of UV light energy. In some cases, the resulting molecule may undergo further biological reaction of toxicological relevance related especially to the photo-carcinogenicity resulting from photo-genotoxicity. However, no regulatory requirements have been issued with the exception of guideline issued by the Scientific Committee of Cosmetology, Commission of the European Communities (SCC/EEC) on the testing of sunscreens for their photo-genotoxicity. Thus, the objectives of this study are to investigate the utility of photo-Ames assay for detecting photo-mutagens, and to evaluate its ability to predict rodent photo-carcinogenicity. Photo-Ames assay was performed on five test substances that demonstrated positive results in photo-carcinogenicity tests: 8-methoxypsoralen (photoactive substance that forms DNA adducts in the presence of ultraviolet A irradiation), chlorpromazine (an aliphatic phenothiazine an a-adr-energic blocking agent), lomefloxacin (an antibiotic in a class of drugs called fluoroquinolones), anthracene (a tricyclic aromatic hydrocarbon a basic substance for production of anthraquinone, dyes, pigments, insecticides, wood preservatives and coating materials) and retinoic acid (a retinoid compound closely related to vitamin A). Out of 5 test substances, 3 showed a positive outcome in photo-Ames assay. With this limited data set, an investigation into the predictive value of this photo-Ames test for determining the photo-carcinogenicity showed that photo-Ames assay has relatively low sensitivity (the ability of a test to predict carcinogenicity). Thus, to determine the use of in vitro genotoxicity tests for prediction of carcinogenicity,' several standard photo-genotoxicity assays should be compared for their suitability in detecting photo-genotoxic compounds.

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Detection of 8-Hydroxy-2' -Deoxyguanosine in Human Peripheral Blood Leukocytes and Stomach Tissues (사람 백혈구 및 위 조직중의 8-Hydroxy-2'-Deoxyguanosine 측정에 관한 연구)

  • Kang Ho Il;Eom Mi Ok;Park Mi Sun;Ryeom Tai Kyung;Jee Seung Wan;Jeon Hea Myung;Kim Ok Hee
    • Environmental Mutagens and Carcinogens
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    • v.25 no.1
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    • pp.13-18
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    • 2005
  • In the present study, we have measured 8-hydroxy-2'-deoxyguanosine in DNA of stomach cancers, adjacent stomach cancer tissues, normal stomach tissues and peripheral blood leukocytes of the same stomach cancer patients (n = 48) to investigate their etiological association with gastric cancer and possibility whether peripheral blood leukocytes can use surrogate marker for early stomach cancer diagnosis by HPLC/ECD system. In normal stomach tissues, we found that 8-hydroxy-2'-deoxyguanosine levels in tissues infected with Helicobacter pylori were 1.4 fold higher than those in tissues without infected with Helicobacter pylori. However, in adjacent stomach cancer tissues, we found that 8-hydroxy-2'-deoxyguanosine levels in tissues infected with Helicobacter pylori were 1.5 fold lower than those in tissues without infected with Helicobacter pylori. In stomach cancer tissues, 8-hydroxy-2'-deoxyguanosine levels in tissues infected with Helicobacter pylori were not significantly different from those in tissues without infected with Helicobacter pylori. In Helicobacter pylori-negative specimens, 8­hydroxy-2'-deoxyguanosine levels of adjacent stomach cancer tissues were found to be significantly higher than those of normal stomach and cancer tissues. The 8-hydroxy-2'-deoxyguanosine levels of female were 1.7 fold higher than those of male in peripheral blood leukocytes of the same stomach cancer patients. The 8-hydroxy-2'­deoxyguanosine levels in Helicobacter pylori-negative specimens among adjacent stomach cancer tissues were found to be reversely correlated with those in peripheral blood leukocytes, suggesting that 8-hydroxy-2'-deox­yguanosine in peripheral blood leukocytes may not use as surrogate marker for the early diagnosis of human stomach cancer.

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Effects of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin (TCDD) on Gene Expression in Mouse Skin Carcinogenesis (마우스 피부암 발생과정에 있어서 2,3,7,8-Tetrachlorodibenzo-p­Dioxin (TCDD) 처리에 의한 유전자발현 변화 연구)

  • Ryeom Tai Kyung;Kim Ok Hee;Kong Mi Kyung;Park Mi Sun;Jee Seung Wan;Eom Mi Ok;Kang Ho Il
    • Environmental Mutagens and Carcinogens
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    • v.25 no.1
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    • pp.40-46
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    • 2005
  • 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) displays high toxicity in animals and has been implicated in human carcinogenesis. Although the mechanism of carcinogenesis by TCDD is unclear, it is considered to be a non-genotoxic compound and tumor promoter. In our experiment, we investigated the effects of TCDD on gene expression in mouse skin carcinogenesis. We used cDNA microarray to detect the differential gene expression in tumors induced in hairless mouse skin by MNNG plus TCDD protocol. We found that erb-2, c-ets2 and p27$^{kip1}$ were significantly up-regulated, but TNFR2, AKT-l, integrin $\beta$l, maspin, IGF-l, c-raf-l, Rb were significantly down-regulated, in tumor region, respectively. We also found that the expression of 53 genes involved in cen cycle, signal transduction, apoptosis, adhesion molecule, angiogenesis, and invasion, were changed two fold more, in tumor surrounding region. These data suggest that TCDD alters the expression of a large array of genes involved in apoptosis, cytokine production and angiogenesis in mouse skin carcinogenesis.

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Protective Effect Naringin on Carbon Tetrachloride Induced Hepatic Injury in Mice (나린진(Naringin)의 $CCl_4$에 의한 급성 간독성 보호효과)

  • Chae, Soo-Chul;Kho, Eun-Gyeong;Choi, Seung-Hyun;Ryu, Geun-Chang
    • Environmental Analysis Health and Toxicology
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    • v.23 no.4
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    • pp.325-335
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    • 2008
  • The protective effects of the Naringin, on carbon tetrachloride ($CCl_4$)-induced hepatotoxicity and the possible mechanisms involved in this protection were investigated in mice. Pretreatment with Naringin prior to the administration of $CCl_4$ significantly prevented an increase in serum alanine, aspartate aminotransferase activity and hepatic lipid peroxidation in a dose-dependent manner. In addition, pretreatment with Naringin also significantly prevented the depletion of glutathione (GSH) content in the livers of $CCl_4$-induced mice. However, reduced hepatic glutathione levels was unaffected by treatment with Naringin alone. In addition, Naringin prevented $CCl_4$-induced apoptosis and necrosis, as indicated by a liver DNA laddering. To determine whether caspase-8,-3 pathway involved in $CCl_4$-induced acute liver injury, caspase-8, -3 activities were tested by ELISA. Naringin attenuated $CCl_4$induced caspase-8, -3 activities in mouse livers. $CCl_4$-induced hepatotoxicity was also prevented, as indicated by a liver histopathologic study. The effects of Naringin on the cytochrome P450 (CYP) 2E1, the major isozyme involved in $CCl_4$ were also investigated. Treatment of mice with Naringin resulted in a significant decrease of the CYP2E1-dependent hydroxyl at ion and aniline in a dose-dependent manner. These findings suggest that protective effects of Naringin against the $CCl_4$-induced hepatotoxicity may be due to its ability to block CYP2E1-mediated $CCl_4$ bioactivation and that is also protects against caspase-8, -3 pathway mediated apoptosis.

Characterization and Antifungal Activity from Soilborne Streptomyces sp. AM50 towards Major Plant Pathogens

  • Jang, Jong-Ok;Lee, Jung-Bok;Kim, Beam-Soo;Kang, Sun-Chul;Hwang, Cher-Won;Shin, Kee-Sun;Kwon, Gi-Seok
    • Korean Journal of Environmental Agriculture
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    • v.30 no.3
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    • pp.346-356
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    • 2011
  • BACKGROUND: Chemical fungicides not only may pollute the ecosystem but also can be environmentally hazardous, as the chemicals accumulate in soil. Biological control is a frequently-used environment-friendly alternative to chemical pesticides in phytopathogen management. However, the use of microbial products as fungicides has limitations. This study isolated and characterized a three-antifungal-enzyme (chitinase, cellulase, and ${\beta}$-1,3-glucanase)-producing bacterium, and examined the conditions required to optimize the production of the antifungal enzymes. METHOD AND RESULTS: The antifungal enzymes chitinase, cellulase, and ${\beta}$-1,3-glucanase were produced by bacteria isolated from an sawmill in Korea. Based on the 16S ribosomal DNA sequence analysis, the bacterial strain AM50 was identical to Streptomyces sp. And their antifungal activity was optimized when Streptomyces sp. AM50 was grown aerobically in a medium composed of 0.4% chitin, 0.4% starch, 0.2% ammonium sulfate, 0.11% $Na_2HPO_4$, 0.07% $KH_2PO_4$, 0.0001% $MgSO_4$, and 0.0001% $MnSO_4$ at $30^{\circ}C$. A culture broth of Streptomyces sp. AM50 showed antifungal activity towards the hyphae of plant pathogenic fungi, including hyphae swelling and lysis in P. capsici, factors that may contribute to its suppression of plant pathogenic fungi. CONCLUSION(S): This study demonstrated the multiantifungal enzyme production by Streptomyces sp. AM50 for the biological control of major plant pathogens. Further studies will investigate the synergistic effect, to the growth regulations by biogenic amines and antifungal enzyme gene promoter.

Diagnostic Method for the Detection of JC Polyomavirus Using Loop-mediated Isothermal Amplification (등온증폭법을 이용한 고감도 JC polyomaviruses 진단법 개발)

  • Cho, Kyu Bong
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.4
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    • pp.414-419
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    • 2019
  • JC polyomavirus (JCPyV) is a human pathogenic virus belonging to the family Polyomaviridae, a viral group containing dsDNA nucleic acid. A recent recommendation is to apply the presence of JCPyV as a fecal indicator for water contamination in environments like sewage, and techniques to monitor JCPyV in water are being proposed. To date, the conventional PCR system has been applied as a diagnostic method for detecting JCPyV. There is a need for a more rapid and sensitive JCPyV diagnostic detection method in clinical and environmental samples. In this study, we developed a loop-mediated isothermal amplification (LAMP) primer set for the detection of JCPyV. Our results indicate that the LAMP method using a specific primer set shows about 10-fold higher detection sensitivity than the conventional PCR system. The effectiveness of the LAMP method developed in this study has been validated by PCR product digestion using the HaeIII restriction enzyme. We, therefore, propose that the LAMP method using a specific primer set can be applied as a rapid and sensitive detection method for monitoring JCPyV in clinical and environmental samples.

Analysis of gene expression in red maple (Acer rubrum) and trembling aspen (Populus tremuloides) populations from a mining region

  • Kalubi, K.N.;Michael, P.;Omri, A.
    • Genes and Genomics
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    • v.40 no.11
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    • pp.1127-1136
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    • 2018
  • The Greater Sudbury Region has been known as one of the most ecologically disturbed areas in Canada for the past century. Plant adaptation to environmental stressors often results in modifications in gene expression at the transcriptional level. The main objective of the present study was to compare the expression of genes associated with nickel resistance in Acer rubrum and Populus tremuloides growing in areas contaminated and uncontaminated with metals. Primers targeting Nramps4, Nas 3, At2G, MRP4 and alpha-tubulin genes were used to amplify cDNA of both species. The expression of the At2G gene, was $2{\times}$ and $9{\times}$ higher in P. tremuloides than in A. rubrum for St. Charles (uncontaminated site) and Kelly Lake (metal contaminated site), respectively. There was a much smaller difference between the two species for the Nramps 4 gene as its expression was $2.5{\times}$ and $3{\times}$ higher in P. tremuloides compared to A. rubrum from St. Charles and Kelly Lake, respectively. The same trend was observed for the MRP4 gene whose expression was $2{\times}$ and $14{\times}$ higher in P. tremuloides than in A. rubrum from St. Charles and Kelly Lake, respectively. For the Nas 3 gene, the expression was similar in both sites. This gene was upregulated $11{\times}$ and $10{\times}$ in P. tremuloides compared to A. rubrum in samples from St. Charles and Kelly Lake, respectively. In general, no significant difference was observed between the metal contaminated and uncontaminated sites for gene expression. In depth analysis revealed that AT2G and MRP4 genes were significantly down regulated in A. rubrum from the metal contaminated sites compared to those from uncontaminated areas, but environmental factors driving this differential gene expression couldn't be established.

Isolation and Identification of Postharvest Spoilage Fungi from Mulberry Fruit in Korea

  • Kwon, O-Chul;Ju, Wan-Taek;Kim, Hyun-Bok;Sung, Gyoo-Byung;Kim, Yong-Soon
    • Korean Journal of Environmental Agriculture
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    • v.37 no.3
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    • pp.221-228
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    • 2018
  • BACKGROUND: Spoilage fungi can reduce the shelf life of fresh fruits and cause economic losses by lowering quality. Especially, mulberry fruits have high sensitivity to fungal attack due to their high water content (> 70%) and soft texture. In addition, the surface of these fruits is prone to damage during harvesting and postharvest handling. However, any study on postharvest spoilage fungi in mulberry fruit has not been reported in Korea. This study aimed to examine the spoilage fungi occurring in mulberry fruits during storage after harvest. METHODS AND RESULTS: In this study, we isolated postharvest spoilage fungi from mulberry fruits stored in refrigerator (fresh fruits) and deep-freezer (frozen fruits) and identified them. In the phylogenetic analysis based on comparisons of the ITS rDNA sequences, the 18 spoilage fungi isolated from mulberry fruits and the 25 reference sequences were largely divided into seven groups that were subsequently verified by high bootstrap analysis of 73 to 100. Alternaria spp. including A. alternate and A. tenuissima, were the most frequently isolated fungi among the spoilage isolates: its occurrence was the highest among the 18 isolates (38.9%). CONCLUSION: The findings of this study will be helpful for increasing the shelf life of mulberry fruits through the application of appropriate control measures against infection by spoilage fungi during storage.

Differentiations between the viable but nonculturable (VBNC) or dead state of Edwardsiella tarda by ethidium monoazide (EMA) treatment-PCR (Ethidium monoazide (EMA) - PCR 법을 이용한 비배양성 생존 상태(VBNC)의 Edwardsiella tarda 검출)

  • Kang, Nam I;Kim, Eunheui
    • Journal of fish pathology
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    • v.31 no.2
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    • pp.93-99
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    • 2018
  • Edwardsiella tarda predominantly causes edwardsiellosis in fish at high temperature, but is rarely isolated from water when water temperature is low. However, E. tarda is viable but nonculturable (VBNC) in low water temperature, but it can be revived when water temperature rises and cause disease to fish. Therefore, in order to prevent disease, it is very important to identify pathogens that are in the VBNC state in environmental water. In this study, E. tarda cells in the VBNC state were detected by the ethidium monoazide (EMA)-PCR method using the low-temperature oligotrophic sea water microcosm obtained by inoculation of E. tarda at a concentration of $10^8CFU/ml$. In order to distinguish between live and dead bacteria in E. tarda, each sample was treated with EMA at different concentrations, photoactivated with a 500 W halogen lamp, and PCR was performed with E. tarda specific primer. At the concentration of $10^7CFU/ml$ bacterium, DNA amplification was observed only in the live cells when treated with $60{\mu}g/ml$ of EMA, and smaller amounts of live cells could be distinguished from dead cells by adjusting the EMA concentration. In addition, the VBNC cells of E. tarda in the oligotrophic low temperature seawater microcosm were estimated to be in the range of $10^4{\sim}10^5CFU/ml$ by EMA-PCR. Therefore, it is possible to detect VBNC cells that will act as potential pathogens in environmental water using EMA-PCR method, and quantitative confirmation using concentration change is also possible.

Complete genome sequence of Lactobacillus koreensis 26-25, a ginsenoside converting bacterium, isolated from Korean kimchi (김치에서 분리한 진세노사이드 전환 능력이 있는 Lactobacillus koreensis 26-25의 유전체 서열 분석)

  • Kim, Ju-Hyeon;Liu, Qing-Mei;Srinivasan, Sathiyaraj;Kim, Myung Kyum;Kim, Sang Yong;Wee, Ji-Hyang;Im, Wan-Taek
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.477-479
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    • 2018
  • A Gram-positive, rod-shaped, ivory colored, and motile, Lactobacillus koreensis 26-25 was isolated from Korean kimchi. Strain 26-25 showed the ability of conversion from major ginsenosides into minor ginsenosides for which whole genome was sequenced. The whole genome sequence of Lactobacillus koreensis 26-25 consisted of one circular chromosome comprised of 3,006,812 bp, with a DNA G + C content of 49.23%. The whole genome analysis of strain 26-25 showed many glycosides hydrolase genes, which may contribute to identify the genes responsible for transformation of major ginsenosides into minor ginsenosides for its high pharmacological effects.