• Title/Summary/Keyword: Environmental DNA

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Diversity and its Characteristics of Life Phenomenon (생명현상의 다양성과 특징에 관한 고찰 - 보건의료에 미치는 영향을 중심으로 -)

  • Lee, Sun-Dong
    • Journal of Society of Preventive Korean Medicine
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    • v.14 no.1
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    • pp.1-12
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    • 2010
  • This research was intended to delve into the diversity of life phenomenon and its characteristics. First of all, this research gave real examples to compare the differences in men's health, disease, and longevity in order to confirm the existence of diversity of life phenomenon. In addition, it also studied the process and mechanism of manifestation of life phenomenon, as well as the influence and problems of existing studies' results and implications. The results are as follow. 1. Differences in health, diseases, and longevity were very big and diverse in researches on different races, nations, ages, socioeconomic status, positions, and even (monozygotic) twins. 2. The basic foundation of all organisms is DNA, and environmental factors change DNA methylation and the structure of chromatin by constantly influencing DNA. Due to this, the manifestation, control, and phenotype of DNA change, resulting in diversified life phenomenon. Therefore, it is the environmental factors, not DNA, that has more influence on the diversity of life. 3. Looking at available studies, the most reasonable perspective on human requires focusing on the diversity of life phenomenon, holistic thinking, and reversible change instead of irreversible determinism. Considerable differences in life phenomenon between entities require a change in malformed perspective on life. Public health and medicine deals with live human beings, a more precise and accurate perspective on life is very important. Because management methods of health and disease, such as structure and approaches of medical research, prevention and cure, must be different by life perspectives.

A New Record of Euonymus hamiltonianus Wall. in Dokdo: Using Molecular Markers of Nuclear and Plastid DNA (독도의 미기록 식물 참빗살나무: 핵과 엽록체 DNA의 분자마커 이용)

  • Song, Im-Geun;Park, Seon-Joo
    • Korean Journal of Environmental Biology
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    • v.32 no.1
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    • pp.88-94
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    • 2014
  • The present scientific scenario shows that, the long-distance dispersal of any species plays a major role in population as well as the flow of individual between populations. However, newly recorded plant at Dokdo was confirmed by using morphological and molecular markers of nrDNA and cpDNA. This scientific study has identified the plant species was as a Euonymus hamiltonianus Wall. (Celastraceae). And after careful studies, the laboratory results shows that the distribution of tree species at Dokdo as an Oceanic Island is a meaningful thing because of the inflow of plant in adverse condition of environment such as a shallow soil layers and steep terrain. So it is necessary to continuous monitor for the inflow of non-indigenous plant into Dokdo.

Evaluation of DNA damage in Pesticide Sprayers using Single Cell Gel Electrophoresis (단세포전기영동법(single Cell Gel Electrophoresis Assay)을 이용한 농약 살포자의 DNA손상 평가)

  • 이연경;이도영;이은일;이동배;류재천;김해준;설동근
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.128-134
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    • 2001
  • Single cell gel electrophoresis (SCGE) assay, also called comet assay, is a rapid and sensitive method to detect DNA damage in single cell level. To evaluate the DNA damage of lymphocytes of pesticides sprayers, SCGE assay was carried out for 50 pesticides sprayer and 58 control subjects. They were interviewed with structured questionnaire to get the information about the kinds and amount of pesticide. Insecticides and fungicides were predominant among pesticides. Major components of pesticides were organophosphorus, organosulfate, cartap, carbamates, and triazole. Sprayed pesticides were classified into two groups. Group I included organophosphorus, organoarsenic, organotin, tetrazine, triazole and gramoxone, which were known to cause DNA damages. Group II pesticide were carbamates, surfactants, organosulfates, etc., which were not found as DNA damaging agents in scientific documents. Olive tail moments of 100 lymphocytes were measured by KOMET 3.1 program for each person. The means of tail moments were compared between farmers exposed to pesticides and control subjects. Farmers showed higher tail moments than control subjects (2.07$\pm$1.40 vs 1.53$\pm$0.77, p<0.05). The means of tail moments also were compared among group I sprayers (n=36), group II sprayers (n=24) and, control subject, and the means or tail moments were 3.4s$\pm$3.2o, 2.66$\pm$2.20 and 1.53$\pm$0.77 respectively. The difference between means of group I sprayers and controls was statistically significant (p<0.05). In conclusion, this study showed higher DNA damage in farmers exposed to pesticides than control subjects, and comet assay could be useful as a biological monitoring method of genotoxic pesticides for farmers.

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Optimal Conditions of Single Cell Gel Electrophoresis (Comet) Assay to detect DNA single strand breaks in Mouse Lymphoma L5178Y cells

  • Ryu, Jae-Chun;Kwon, Oh-Seung;Kim, Hyung-Tae
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.89-94
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    • 2001
  • Recently, single cell gel electrophoresis, also known as comet assay, is widely used for the detection and measurement of DNA strand breaks in vitro and in vivo in many toxicological fields such as radiation exposure, human monitoring and toxicity evaluation. As well defined, comet assay is a sensitive, rapid and visual method for the detection of DNA strand breaks in individual cells. Briefly, a small number of damaged cells suspended in a thin agarose gel on a microscope slide were lysed, unwinded, electrophoresed, and stained with a fluorescent DNA binding dye. The electric current pulled the charged DNA from the nucleus such that relaxed and broken DNA fragments migrated further. The resulting images which were subsequently named for their appearance as comets, were measured to determine the extent of DNA damages. However, some variations could be occurred in procedures, laboratories's conditions and kind of cells used. Hence, to overcome and to harmonize these matters in comet assay, International Workshop on Genotoxicity Test Procedure (IWGTP) was held with several topics including comet assay at Washington D.C. on March, 1999. In spite of some consensus in procedures and conditions in IWGTP, there are some problems still remained to be solved. In this respect, we attempted to set the practical optimal conditions in the experimental procedures such as lysis, unwinding, electrophoresis and neutralization conditions and so on. First of all, we determined optimal lysis and unwinding time by using 150 $\mu$M methyl methanesulfonate (MMS) which is usually used concentration. And then, we determined optimal positive control concentrations of benzo(a)pyrene (BaP) and MMS in the presence and absence of S9 metabolic activation system, respectively.

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Genotoxic Effects of Diesel Exhaust Particle Extract in NIH/3T3 Cells (디젤분진이 체세포에서의 DNA 손상에 미치는 영향)

  • Heo Chan;Kim Nam Yee;Chung Kyu-Hyuek;Moon Chang-Kiu;Heo Moon Young
    • Environmental Analysis Health and Toxicology
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    • v.19 no.4
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    • pp.335-344
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    • 2004
  • Diesel exhaust particle (<2.5 ${\mu}{\textrm}{m}$, DEP$_{2.5}$) is known to be probarbly carcinogenic (IARC group 2A). DEP$_{2.5}$ contains organic compounds such as polycyclicaromatic hydrocarbon (PAH), heterocyclic compounds, phenols, and nitroarenes. Reactive oxygen species (ROS) are generated by DEP$_{2.5}$ without any biological activation system. Therefore, an alternative mechanism by which DEP$_{2.5}$ could be carcinogenic is known by the generation of oxidative DNA damage. The aim of this study was to investigate genotoxic effects of DEP$_{2.5}$ using single cell gel electrophoresis. In order to evaluate the mechanisms of DEP$_{2.5}$ genotoxicity, the rat micro-some mediated and DNA repair enzyme treated comet assays together with routine comet assay were performed. DEP$_{2.5}$ was collected from diesel engine bus and dichloromethane extract was obtained. The organic extract of DEP$_{2.5}$ revealed DNA damage itself in NIH/3T3 cells. And it showed both oxidative and microsome mediated DNA damages. Vitamin C as an model antioxidant reduced DNA damage in endonuclase III treated comet assay. One of flavonoid, galangin as a CYP1A1 inhibitor reduced DNA damage in the presence of S-9 mixture. Our results show that DEP$_{2.5}$ are genotoxic and a great source of oxidative stress, but antioxidants can significantly reduce oxidative DNA damages. And DEP$_{2.5}$ may contain indirect mutagens which can be inhibited by CYP inhibitors.d by CYP inhibitors.

Development of a Highly Efficient Isolation Protocol for Mitochondrial DNA and RNA Using Small Scale Plant Tissues (식물의 초경량 조직을 이용한 미토콘드리아의 DNA와 RNA 정제)

  • Kim Kyung-Min;Lim Yong-Suk;Shin Dong-Ill;Sul Ill-Whan
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.240-244
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    • 2006
  • We present a fast and simple protocol for purification of mitochondria, mitochondrial DNA, and RNA from small amounts of tomato leaves. This method uses a high ionic strength medium to isolate mitochondria and extract mitochondrial DNA and RNA from a single preparation and is easily adaptable to other plant species. Mitochondria was confirmed by MitoTracker. The mitochondrial DNA was not contaminated by plastid DNA, was successfully used for PCR. Similarly, the isolated mitochondrial RNA was not contaminated only slightly contaminated (leaves) by plastid RNA. RNA prepared according to our method was acceptable for RT-PCR analysis

A Thermodynamic Study on the Interaction of Quinolone Antibiotics and DNA

  • Lee, Byung-Hwa;Yeo, Ga-Young;Jang, Kyeung-Joo;Lee, Dong-Jin;Noh, Sang-Gyun;Cho, Tae-Sub
    • Bulletin of the Korean Chemical Society
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    • v.30 no.5
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    • pp.1031-1034
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    • 2009
  • Fluorescence of quinolones including norfloxacin, ciprofloxacin and S- and R-ofloxacin is quenched upon association with single and double-stranded DNA (ss- and ds-DNA). The ratios of fluorescence intensity in the presence of DNA to its absent were plotted with respect to the DNA concentration to construct the Stern-Volmer plot. The slope of the Stern-Volmer plot become larger as the temperature is lowered, ensuring that the fluorescence quenching is static process, i.e., the fluorescence is quenched by formation of the non-fluorescent complex between quinolone and DNA. In the static quenching mechanism, the quenching constant which is equivalent to the slope of the Stern-Volmer plot, is considered as the equilibrium constant for the association of quinolones and DNA. From the temperature-dependent equilibrium constant, ${\Delta}H^0\;and\;{\Delta}S^0$ was obtained using the van’t Hoff relation. In general, association of the quinolone with ds- as well as ss-DNA is energetically favorable (an exothermic) process while the entropy change was unfavorable. Due to the steric effect of the substituents, the effect of the quinolone ring is smaller on the ss-DNA compared to ds-DNA.

Analysis of genes expressed during pepper-Phytophthora capsici interaction

  • Park, Woobong;Jeon, Myoung-Seung;Kim, Yean-Hee;Park, Eun-Woo;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.86-86
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    • 2003
  • Phytophthora capsici is a pathogen on several economically important crops including pepper. In pepper growing areas in Korea, Phytophthora blight caused by p. capsici has been considered as the most serious problem in pepper production. The Oomycete attacks the roots, stems, leaves and fruits of the plant. To understand the molecular mechanisms involved in the disease development, the genes expressed doting pepper p. capsici interaction were explored by analyzing expressed sequence tags (ESTs). A complementary DNA (cDNA) library was constructed from total RNA extracted from pepper leaves challenged with p. capsici for 3 days resulting in early stage of symptom development. The comprehensive analysis on the single pass sequencing of over 4000 randomly selected cDNA clones with contig assembly, unique gene extraction, sequence comparison, and functional categorizing will be presented with an emphasis on the genes involved in plant defense and pathogenicity during disease development of the pepper Phytophthora blight.

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Gene Expression Profiles of Dibutyl Phthalate and 17$\beta$-Estradiol using cDNA microarray in MCF 7 Human Breast Cancer Cell Line

  • Ryu, Jae-Chun;Kim, Hyung-Tae;Kim, Youn-Jung
    • Environmental Mutagens and Carcinogens
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    • v.22 no.4
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    • pp.274-278
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    • 2002
  • Phthalates, suspected endocrine disruptor, are plasticizer and solvent used in industry, and some phthalates are known as potential carcinogen. Most common human exposure to this compounds may occur with contaminated food. It may migrate into food from plastic wrap or may enter food from general environmental contamination, and it has become widespread environmental pollutants, thus leading to a variety of phthalates that possibly threaten the public health. Dibutyl phthalate (DBP) may playa part of cell proliferator, which mediates changes in gene expression and the metabolism of xenobiotics. An understanding of the role of DBP in modulating gene regulation should provide insight regarding mechanisms of DBP induced xenoestrogenic impact. To elucidate the type of genes that are associated with estrogenic activity induced by DBP at the dose (10$^{-8}$ M) appeared proliferating effects, the pattern of gene expression in MCF7 cells was compared between 17$\beta$-estradiol and DBP exposure in the cDNA microarray. From the results, it showed some differences of gene expression patterns between MCF7 cells treated with 17$\beta$-estradiol and DBP, and also DBP shows estrogenic potential with changes in estrogen-related gene expression levels.

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Evaluation of DNA Microarray Approach for Identifying Strain-Specific Genes

  • Hwang, Keum-Ok;Cho, Jae-Chang
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1773-1777
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    • 2006
  • We evaluated the usefulness of DNA microarray as a comparative genomics tool, and tested the validity of the cutoff values for defining absent genes in test genomes. Three genome-sequenced E. coli strains (K-12, EDL933, and CFT073) were subjected to comparative genomic hybridization with DNA microarrays covering almost all ORFs of the reference strain K-12, and the microarray results were compared with the results obtained from in silico analyses of genome sequences. For defining the K-12 ORFs absent in test genomes (reference strain-specific ORFs), we applied and evaluated the cutoff level of -1. The average sequence similarity between ORFs, to which corresponding spots showed a log-ratio of>-1, was $96.9{\pm}4.8$. The numbers of spots showing a log-ratio of <-1 (P<0.05, t-test) were 90 (2.5%) and 417 (10.6%) for the EDL933 genome and the CFT073 genome, respectively. Frequency of false negatives (FN) was ca. 0.2, and the cutoff level of -1.3 was required to achieve the FN of 0.1. The average sequence similarity of the false negative ORFs was $77.8{\pm}14.8$, indicating that the majority of the false negatives were caused by highly divergent genes. We concluded that the microarray is useful for identifying missing or divergent ORFs in closely related prokaryotic genomes.