• Title/Summary/Keyword: environmental DNA

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Molecular Identification of the Toxic Alexandrium tamiyavanichii (Dinophyceae) by the Whole-cell FISH Method

  • Kim Choong-Jae;Yoshimatsu Sada-Akfi;Sako Yoshihiko;Kim Chang-Hoon
    • Fisheries and Aquatic Sciences
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    • v.7 no.4
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    • pp.175-183
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    • 2004
  • The dinoflagellate Alexandrium tamiyavanichii Balech, a producer of toxins causing paralytic shellfish poisoning (PSP), has recently been considered as one of main organisms responsible for toxication of shellfish in Japan. In this study, A. tamiyavanichii was subjected to a molecular phylogenetic analysis inferred from 28S rDNA D1-D2 sequences and a species-specific LSU rRNA-targeted oligonucleotide DNA probe was designed to identify A. tamiyavanichii using the whole cell-FISH (fluorescence in situ hybridization). The sequences of the 28S rDNA D1-D2 region of A. tamiyavanichii showed no difference from A. cohorticular AF1746l4 (present name A. tamiyavanichii) and formed a distinct clade from the 'tamarensis species complex'. The probe, TAMID2, reacted specifically with A. tamiyavanichii cultured cells, without any cross-reaction with other species belonging to the same genus, including A. tamarense, A. catenella, A. affine, A. fraterculus, A. insuetum and A. pseudogonyaulax. In a test of cross-reactivity with a field sample, TAMID2 reacted consistently with only A. tamiyavanichii, indicating that the present protocol involving the TAMID2 probe might be useful for detecting toxic A. tamiyavanichii in a simple and rapid manner.

Genotyping of Six Pathogenic Vibrio Species Based on RFLP of 16S rDNAs for Rapid Identification

  • Yoon, Young-Jun;Im, Kyung-Hwan;Koh, Young-Hwan;Kim, Seong-Kon;Kim, Jung-Wan
    • Journal of Microbiology
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    • v.41 no.4
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    • pp.312-319
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    • 2003
  • In an attempt to develop a method for rapid and accurate identification of six Vibrio species that are clinically important and most frequently detected in Korea, 16S rDNA restriction fragment length polymorphism (RFLP) of Vibrio type strains, as well as environmental isolates obtained from the Korean coastal area, was analyzed using ten restriction endonucleases. Digestion of the 16S rDNA fragments amplified by polymerase chain reaction (PCR) with the enzymes gave rise to 2~6 restriction patterns for each digestion for 47 Vibrio strains and isolates. An additional 2~3 restriction patterns were observed for five reference species, including Escherichia coli, Aeromonas hydrophila, A. salmonicida, Photobacterium phosphoreum, and Plesiomonas shigelloides. A genetic distance tree based on RFLP of the bacterial species correlated well with that based on 16S rDNA sequences. The very small 16S rDNA sequence difference (0.1%) between V. alginolyticus and V. parahaemolyticus was resolved clearly by RFLP with a genetic distance of more than 2%. RFLP variation within a species was also detected in the cases of V. parahaemolyticus, V. proteolyticus, and V. vulnificus. According to the RFLP analysis, six Vibrio and five reference species were assigned to 12 genotypes. Using three restriction endonucleases to analyze RFLP proved sufficient to identify the six pathogenic Vibrio species.

Carcinogenic Potentials of HPV-16 and NNK in Human in Vitro Model (인체 세포 모델을 이용한 HPV-16과 NNK의 발암 잠재력에 관한 연구)

  • 양재호;이세영
    • Toxicological Research
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    • v.12 no.2
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    • pp.271-275
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    • 1996
  • Carcinogenic potential of HPV-16 DNA and NNK in a human keratinocyte cell line was assessed to study effects of viral-chemical interaction. Human cells were transfected with HPV-16 DNA and 6 clonal cell lines were subsequently obtained. Clonal line-3 and 6 at passage 7 showed characteristics of tumor cells such as increases of saturation density, soft-agar colony formation, cell aggregation and foci appearance. Among cells treated with 1$\mu M$, 10$\mu M$, 100$\mu M$ or 1 mM of NNK for 4 weeks, 100$\mu M$ treatment showed most tumorigenic characteristics at passage 7. These results indicate that either HPV-16 or NNK alone is tumorigenic in this in human in vitro model. When cells transfected with HPV-16 were subsequently exposed by 100 uM NNK for 4 weeks, all the clonal cells except clone-1 showed higher levels of tumor cell characteristics than HPV-16 DNA or NNK exposure alone. Clonal line-6, the most tumorigenic cells, showed higher transcriptional level of fibronectin and lower level of TGF-$\beta_1$, as compared to control cells, suggesting that alteration of growth factor or extracellular matrix may play a role in carcinogenesis process induced by HPV-16 and NNK. Taken together, the present study indicates that viral-chemical interactions between HPV-16 DNA and NNK enhance carcinogenic potentials of human cells and implies that smoking among people infected with human papillomavirus may pose an additional risk of causing cancer.

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Inhibitory Effect of Vitamin C on Mutagenicity of 6-Sulfooxymethylbenzo[a]pyrene

  • Cho, Young-Sik;Hong, Sun-Taek;Chung, An-Sik
    • Toxicological Research
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    • v.12 no.1
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    • pp.21-27
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    • 1996
  • Vitamin C has been well known to be a potential chemopreventive agent for several toxic compounds. It reduced the mutation frequencies of 6-sulfooxymethylbenzo[a]pyrene (SMBP) and 6hydroxymethylbenzo[a]pyrene (HMBP) in Salmonella typhimurium TA98 and TA100, indicating that corbic acid affects both frameshift and base-pair substitution mtltations. A similar type of dose-response relationship was shown in the V79 cells, although the inhibitory effect was less dramatic compared with that in S. typhimurium. However, SMBP or HMBP binding to calf thymus DNA was not affected by the presence of vitamin C, suggesting that SMBP seems to be much more reactive to calf thymus DNA than vitamin C. This was supported by migration pattern and fluorescence intensity of SMBP-modified plasmid on the gel. These restilts were not correlated with mutation tests in bacterial and mammalian cell systems. It has been already reported that vitamin C inactivates SMBP through the formation of covalently bound addact. It was found from HPLC analysis that the reaction between vitamin C and SMBP was accomplished within just 5 min and then produced the several products. These findings indicate that the beneficiary of vitamin C is not merely derived from the covalent adducts. On the other hand, the addition of DNA to incubation mixture reduced the amounts of vitamin C adducts while the magnitude of HMBP peak increased, suggesting that DNA accelerates the SMBP hydrolysis to intercept the interaction between SMBP and vitamin C or forms rapidly complex with SMBP.

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Investigation of genomic integration of GX-12, a new anti-HIV DNA vaccine, into host cellular DNA following intramuscular injection in rats

  • Kang, Kyung-Koo;Park, Min-Seul;Ahn, Jun-Kook;Baik, Dae-Hyun;Lee, Dong-Sup;Park, Jae-Hun;Ahn, Byoung-Ok;Kim, Won-Bae
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.11b
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    • pp.199-200
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    • 2002
  • GX-12 is a naked DNA vaccine developed by research team of Dong-A Pharmaceutical Company, Green Cross Company and Genexine for the treatment of HIV infection. It consists of four separate plasmids (pGX10-GE HX, pGX10-dpol JR, pGX10-VN/TV JR, pGX10-hIL-12m), which were constructed by inserting the HIV-1 gag-env, pol, regulatory genes and a human IL-12 mutant gene into pGX10 plasmid vectors.(omitted)

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Four New Records of Parasitoid Wasps of Genus Aleiodes (Hymenoptera: Braconidae: Rogadinae) from South Korea (한국산 미기록 기생벌 Aleiodes (벌목: 고치벌과: 송충살이고치벌아과)속 4종에 대한 보고)

  • Gyeonghyeon Lee;Juhyeong Sohn;Hyeban Namgung;Jongok Lim;Hyojoong Kim
    • Korean journal of applied entomology
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    • v.62 no.1
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    • pp.49-56
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    • 2023
  • We first report 4 unrecorded species belonging to genus Aleiodes Wesmael discovered in South Korea: A. lipwigi, A. guidaae, A. subemarginatus, A. caprinus. In this study, diagnosis, distribution, and illustration, DNA barcode sequence are provided for identification of these species.

Epigenetic Age Prediction of Alzheimer's Disease Patients Using the Aging Clock (노화 시계를 이용한 알츠하이머병 환자의 후성유전학적 연령 예측)

  • Jinyoung Kim;Gwang-Won Cho
    • Journal of Integrative Natural Science
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    • v.16 no.2
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    • pp.61-67
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    • 2023
  • Human body ages differently due to environmental, genetic and pathological factors. DNA methylation patterns also differs depending on various factors such as aging and several other diseases. The aging clock model, which uses these differences to predict age, analyzes DNA methylation patterns, recognizes age-specific patterns, predicts age, and grasps the speed and degree of aging. Aging occurs in everyone and causes various problems such as deterioration of physical ability and complications. Alzheimer's disease is a disease associated with aging and the most common brain degenerative disease. This disease causes various cognitive functions disabilities such as dementia and impaired judgment to motor functions, making daily life impossible. It has been reported that the incidence and progression of this disease increase with aging, and that increased phosphorylation of Aβ and tau proteins, which are overexpressed in this disease and accelerates epigenetic aging. It has also been reported that DNA methylation is significantly increased in the hippocampus and entorhinal cortex of Alzheimer's disease patients. Therefore, we calculated the biological age using the Epi clock, a pan-tissue aging clock model, and confirmed that the epigenetic age of patients suffering from Alzheimer's disease is lower than their actual age. Also, it was confirmed to slow down aging.

Effect of the Magnetized Water Supplementation on Lymphocyte DNA Damage in Mice Treated with Diethylnitrosamine (Diethyl Nitrosamine (DEN) 처리 실험동물에 있어 기간에 따른 자화육각수의 임파구 DNA 손상 개선효과)

  • Lee, Hye-Jin;Jo, Hye-Ryun;Jeon, Eun-Jae;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.43 no.6
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    • pp.570-577
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    • 2010
  • Water gets magnetically charged when it is contacted with a magnet. Although magnetic water products have been promoted since the 1930's, they have received very little recognition due to questionable effectiveness. Diethylnitrosamine (DEN) is a widely occurring nitrosamine that is one of the most important environmental carcinogens primarily inducing tumors of liver. In this study, the effect of magnetized water supplementation on lymphocyte DNA damage in ICR mice treated with DEN was evaluated using the Comet assay. Mice were divided into 3 groups: control, DEN, and DEN + magnetized water group. Fifteen mice were maintained in each group for the entire experimental period of 6, 12 and 18 weeks. Five mice in each group were sacrificed at 6, 12, and 18th weeks, followed by the Comet assay using the blood obtained from heart puncture of the mice. The level of lymphocyte DNA damage reflected by tail moment and other DNA damage indices of tail DNA (%) or tail length of the magnetized water group were significantly decreased after the 6th, 12th and 18th weeks of supplementation compared with the positive control, the DEN group. The relative DNA damage of the magnetized water groups compared to the DEN control group after 6th, 12th, and 18th weeks of supplementation were 42.2%, 40.8%, and 32.9% for DNA in tail, 31.2%, 32.6%, and 21.3% for tail length, and 33.8%, 33.8%, and 24.6% for tail moment, respectively. This is the first report demonstrating that magnetized water may be involved in the lowering effect of the DNA damage in DEN-treated ICR mice. This result suggests that the magnetized water might have minimized the DNA damage by improving the antioxidant status of the mice. However, further studies are needed to characterize the condition of the magnetization and examine the long-term effect of the water product.

Physiological Function of a DNA-Binding Protein from Starved Cells in Combating Diverse External Stresses in Escherichia coli (대장균 세포 내 다양한 외부 스트레스에 대한 DPS 단백질의 생리적 기능)

  • Lee, Joo Hyeong;Cheong, Su Jin;Oh, Hun Taek;Kim, Woe Yeon;Jung, Young Jun
    • Journal of Life Science
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    • v.23 no.4
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    • pp.479-486
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    • 2013
  • The DNA-binding protein from starved cells (DPS), originally identified as a DNA binding protein in Escherichia coli, is known to play an important role in DNA protection. The aim of this study was to evaluate the functional roles of DPS in E. coli against various kinds of external stresses by comparing the properties of wild-type E. coli cells and dps knockout mutant E. coli (${\Delta}dps$) cells. Under various stress conditions, we measured the cell growth of the wild-type E. coli and the dps knockout mutant E. coli (${\Delta}dps$) cells using a UV spectrophotometer. The growth rate of the cells was compared to investigate the functional roles of the DPS protein in E. coli. In comparison to the properties of the wild-type E. coli cells, the dps knockout mutant E. coli (${\Delta}dps$) cells showed highly sensitive phenotypes under various stress conditions, such as heat shock, acidic pH, nutrient deficiency, and different concentrations of reactive oxygen species (ROS), suggesting that DPS plays key roles in E. coli in combating diverse external stresses. The DPS DNA-binding protein in E. coli plays crucial roles in bacterial cell growth and in the protection of the cells from environmental stresses by tightly binding and preserving their DNA molecules.

Distribution Status of Invasive Alien Species (Procambarus clarkii (Girard, 1852)) Using Biomonitoring with Environmental DNA in South Korea (생물모니터링 및 환경유전자(eDNA)를 이용한 침입외래종 미국가재(Procambarus clarkii)의 한국분포 현황)

  • Jung, Sang Woo;Lee, Jae-Ha;Kawai, Tadashi;Kim, Phil-Jae;Kim, SuHwan
    • Korean Journal of Environment and Ecology
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    • v.36 no.4
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    • pp.368-380
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    • 2022
  • Biomonitoring of an invasive alien crayfish species, Procambarus clarkii (Girard, 1852), was performed from February to October 2021, along with environmental DNA analysis, at five locations including Wanju-gun, Hampyeong-gun, Naju-si, Gurye-gun, and Cheongju-si. For the investigation, an umbrella-shaped trap for adults and a hand net for young crayfish were used, and 8 to 10L of freshwater was collected for eDNA analysis. The current status and past distributional records of crayfish in Korea were analyzed along with benthic macroinvertebrates at each survey site. As a result of the investigation, a total of 122 individuals were identified, and Hampyeong-gun recorded the largest number of populations with 59 individuals (48.36%) and the highest environmental DNA (eDNA). The frequency of appearance of P. clarkii was highest in May. The ratio of females to males was 21:5, and the body size was 72.2±21.1mm for female, 80.5±15.6mm for male, and 25.3±9.8mm for young crayfish. P. clarkii introduced into Korea is mainly spreading in the southwest region and it has not been observed in Seoul where there had been a record of appearance in the past. No external symbiosis (Branchiobdellida) of P. clarkii has been identified, and P. clarkii that has appeared in Korea was presumed to be imported from Japan. There are more than eight kinds of exotic crayfish distributed in Korea, and among them, the marbled crayfish (P. virginalis) was identified as a harmful species to the ecosystem of Korea by the Ministry of Environment in 2021. The identified species of benthic macroinvertebrates inhabiting the survey area where P. clarkii has appeared were 69 belonging to 39 families, 15 orders, five classes, and three phyla. Among them, Odonata were the most abundant (16 spp.; 24.62%), followed by Coleoptera (11 spp.; 16.92%) and Hemiptera (11 spp.; 16.92%). In the survey area, one Korean endemic species (Rhoenanthus coreanus), one species (Helophorus auriculatus) classified as Near Threatened (NT) on the Korean Red List, and six species of Crustaceans appeared. In the functional feeding group, the predators appeared predominantly while in habitat oriented group, the climbers appeared to be abundant. It was confirmed that P. clarkii in the survey area prefers an area rich in aquatic vegetation with waterside vegetation, and has high resistance to turbid water quality. The omnivore invader P. clarkii is expected to maintain a competitive relationship with carnivorous benthic macroinvertebrates that are predominant in the same species, and is expected to continue to generate ecosystem disturbance along the food chains.