• Title/Summary/Keyword: environmental DNA

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Naegleria fowleri Induces Jurkat T Cell Death via O-deGlcNAcylation

  • Lee, Young Ah;Kim, Kyeong Ah;Shin, Myeong Heon
    • Parasites, Hosts and Diseases
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    • v.59 no.5
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    • pp.501-505
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    • 2021
  • The pathogenic free-living amoeba Naegleria fowleri causes primary amoebic meningoencephalitis, a fatal infection, by penetrating the nasal mucosa and migrating to the brain via the olfactory nerves. N. fowleri can induce host cell death via lytic necrosis. Similar to phosphorylation, O-linked β-N-acetylglucosamine (O-GlcNAc) glycosylation (O-GlcNAcylation) is involved in various cell-signaling processes, including apoptosis and proliferation, with O-GlcNAc addition and removal regulated by O-GlcNAc transferase and O-GlcNAcase (OGA), respectively. However, the detailed mechanism of host cell death induced by N. fowleri is unknown. In this study, we investigated whether N. fowleri can induce the modulation of O-GlcNAcylated proteins during cell death in Jurkat T cells. Co-incubation with live N. fowleri trophozoites increased DNA fragmentation. In addition, incubation with N. fowleri induced a dramatic reduction in O-GlcNAcylated protein levels in 30 min. Moreover, pretreatment of Jurkat T cells with the OGA inhibitor PUGNAc prevented N. fowleri-induced O-deGlcNAcylation and DNA fragmentation. These results suggest that O-deGlcNAcylation is an important signaling process that occurs during Jurkat T cell death induced by N. fowleri.

Mitochondrial DNA Mutation (3243A→G,1555A→4G,7445A→G) in Noise-Induced (소음성 난청에서의 Mitochondrial DNA A3243G, A1555G, A7445G 돌연변이)

  • Hong Young-Seoub;Nishio Hisahide;Lee Myeong-Jin;Kwak Ki-Young;Hwang Chan-Ho;Shin Dong-Hoon;Kwak Jong-Young;Lee Yong-Hwan;Kim Jong-Min;Kim Joon-Youn
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.913-919
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    • 2004
  • Mitochondrial DNA mutations have been reported in recent years in association with sensorineural hering loss. The purpose of this study is to identify the association between the noise-induced sensorineural hearing loss and the A to G mutation at nucleotide 3243, 1555, 7445 of mitochondrial DNA. Study subjects were established by history and chart review, and audiological and clinical data were obtained. Blood was sampled from 214 normal controls, 102 noise-induced hearing loss, and 28 sensorineural hearing loss. The DNA of these individuals were extracted, and mitochondrial DNA fragments were analyzed by polymerase chain reaction. Subsequently, the coding sequence of mitochondrial DNA 3243, 1555, 7445 were sequenced, and compared to the normal sequence, and all sequence variations were analyzed by restriction enzymes. Mitochondrial DNA mutations $(3243A{\rightarrow}G,\;1555A{\rightarrow}4G,\;7445A{\rightarrow}G)$ were not detected by polymerase chain reactions in any patients with noise-induced hearing loss, sensorineural hearing loss, and normal controls. The DNA sequencing of PCR products did not revealed an A to G substitution at nucleotide 3243, 1555, 7445 of mitochondrial DNA. The noise-induced sensorineural hearing loss was not associated with mitochondrial DNA mutation $(3243A{\rightarrow}G,\;1555A{\rightarrow}4G,\;7445A{\rightarrow}G)$.

Effects of Oxidative DNA Damage Induced by Polycyclic Aromatic Hydrocarbons and Genetic Polymorphism of the Paraoxonase-1 (PON1) Gene on Lung Cancer (다환성 방향족 탄화수소 노출에 의한 DNA 산화적 손상과 Paraoxonase-1(PON1) 유전자 다형성이 폐암 발생에 미치는 영향)

  • Lee, Chul-Ho;Lee, Kye-Young;Choe, Kang-Hyeon;Hong, Yun-Chul;Kim, Yong-Dae;Kang, Jong-Won;Kim, Heon
    • Journal of Preventive Medicine and Public Health
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    • v.38 no.3
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    • pp.345-350
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    • 2005
  • Objectives : Polycyclic aromatic hydrocarbons (PAHs), which are risk factors for lung cancer, have been reported to induce oxidative DNA damage. The paraoxonase (PON) plays a significant role in the detoxification of a variety of organophosphorous compounds, with paraoxonase-1 (PON1) being one of the endogenous free-radical scavenging systems in the human body. The aim of this case-control study was to investigate the effects of PAH exposure, oxidative stress and the Q192R polymorphism of PON1 genes, and their interactions in the carcinogenesis of lung cancer. Methods : One hundred and seventy seven lung cancer patients and 177 age- and sex-matched controls were enrolled in this study. Each subject was asked to complete a questionnaire concerning their smoking habits and environmental exposure to PAHs. The Q192R genotypes of the PON1 gene was examined, and the concentrations of urinary 1-hydroxypyrene (1-OHP), 2-naphthol and 8-hydroxydeoxyguanosine (8-OH-dG) measured. Results : Cigarette smoking was found to be a significant risk factor for lung cancer. The urinary 8-OH-dG level was higher in the patients, whereas the urinary 1-OHP and 2-naphthol levels were higher in the controls. There was a significant correlation between the urinary levels of 8-OHdG and 1-OHP in both the cases and controls. The PON1 polymorphism was associated with an increased risk of lung cancer. Individuals carrying the Q/Q genotype of the PON1 gene were found to be at higher risk of developing lung cancer. There was a significant correlation between the urinary levels of 8-OH-dG and 1-OHP in those with the PON1 Q/Q genotype. Conclusions : These results lead to the conclusion that PAHs would induce oxidative DNA damage, especially in individuals with the PON1 Q/Q genotype. Therefore, people with the PON1 Q/Q genotype would be more susceptible to lung cancer than those with the R/R or Q/R genotypes of the PON1 gene.

Complete genome sequence of Spirosoma aerolatum KACC 17939T, a bacterium related to the DNA repair (DNA 복원에 관련된 박테리아 Spirosoma aerolatum KACC 17939T의 완전한 게놈 서열)

  • Kim, Dong-Uk;Kim, Ju-Young;Kim, Su Jeong;Kim, Min Ji;Lee, Ju Yeon;Kim, Myung Kyum
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.230-232
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    • 2017
  • A Gram-stain-negative, yellow-pigmented bacterial strain, designated Spirosoma aerolatum KACC $17939^T$, was isolated from a biofilm of car air conditioner collected in Republic of Korea. In this study, we report the complete genome sequence of a bacterium Spirosoma aerolatum KACC $17939^T$ obtained using the PacBio RS II platform. The genome comprised of 7,959,595 bp with the G + C content of 48.3%, the genome included 6,640 genes were predicted, among them, 6,471 genes are protein-coding genes.

Change of Microbial Communities in Fermentative Hydrogen Production at Difference Cultivation pHs (혐기성 수소생산 시 운전 pH 변화에 따른 미생물의 군집 변화)

  • Jun, Yoon-Sun;Lee, Kwan-Yong;Cho, Yoon-A;Lee, Tae-Jin
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.12
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    • pp.1239-1244
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    • 2008
  • In this study, PCR-DGGE was conducted to investigate the variations of microbial community according to pH conditions from pH 3 to pH 10 during anaerobic fermentation process of hydrogen production. Maximum hydrogen yield was 1.8 mol $H_2$/mol substrate at pH 5. The microbial growth rate was not proportional to the hydrogen production rate at each pH. Variations of microbial community was observed at each condition from PCR-DGGE experiment of 16s rDNA. Klebsiella was main species of the microbial community. Streptococcus and Clostridium were mainly contributed for hydrogen production.

Next-generation Sequencing for Environmental Biology - Full-fledged Environmental Genomics around the Corner (차세대 유전체 기술과 환경생물학 - 환경유전체학 시대를 맞이하여)

  • Song, Ju Yeon;Kim, Byung Kwon;Kwon, Soon-Kyeong;Kwak, Min-Jung;Kim, Jihyun F.
    • Korean Journal of Environmental Biology
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    • v.30 no.2
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    • pp.77-89
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    • 2012
  • With the advent of the genomics era powered by DNA sequencing technologies, life science is being transformed significantly and biological research and development have been accelerated. Environmental biology concerns the relationships among living organisms and their natural environment, which constitute the global biogeochemical cycle. As sustainability of the ecosystems depends on biodiversity, examining the structure and dynamics of the biotic constituents and fully grasping their genetic and metabolic capabilities are pivotal. The high-speed high-throughput next-generation sequencing can be applied to barcoding organisms either thriving or endangered and to decoding the whole genome information. Furthermore, diversity and the full gene complement of a microbial community can be elucidated and monitored through metagenomic approaches. With regard to human welfare, microbiomes of various human habitats such as gut, skin, mouth, stomach, and vagina, have been and are being scrutinized. To keep pace with the rapid increase of the sequencing capacity, various bioinformatic algorithms and software tools that even utilize supercomputers and cloud computing are being developed for processing and storage of massive data sets. Environmental genomics will be the major force in understanding the structure and function of ecosystems in nature as well as preserving, remediating, and bioprospecting them.

Isolation and Identification of ura5 Gene in Entomopathogenic Fungus, Metarhizium anisopoliae (살충성곰팡이 Metarhizium anisopliae의 ura5 유전자의 분리동정)

  • Park, In-Cheol;Lee, Dong-Kyu;Kang, Sun-Cheol;Hwang, Cher-Won
    • Applied Biological Chemistry
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    • v.40 no.1
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    • pp.30-33
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    • 1997
  • About 250 bp ura5 gene (Orotate phosphoribosyl transferase) fragment was cloned from genomic DNA of entomopathogenic fungus Metarhizium anisopliae by using PCR method. Entire nucleotide sequences of cloned DNA fragment were determined and analysed as compared with other fungus ura5 genes. The amino acid sequence deduced from the nucleotide sequence showed 85.5% homology to ura5 protein of Trichoderma reesei. Using this 250 bp PCR fragment we have isolated full ura5 gene of M. anisopliae by genomic Southern hybridization and the isolated 4.4 kb DNA fragments were mapped by restrictional enzyme.

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Cloning and Characterization of UV-B Inducible Chalcone Synthase from Grape Cell Suspension Culture System and Its Expression Compared with Stilbene Synthase

  • Song, Won-Yong;In, Jun-Gyo;Lim, Yong-Pyo;Park, Kwan-Sam
    • Journal of Photoscience
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    • v.7 no.2
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    • pp.53-58
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    • 2000
  • We performed the cloning of a chalcone synthase (CHS) gene, the key enzyme in the anthocyanin biosynthesis, from the cDNA library constructed with grape suspension cells irradiated UV-B. The PCR fragment was used to cloning the CHS gene. One CHS cDNA clone containing an open reading frame and a partial stilbene synthase (STS)cDNA, the stilbene-type phytoalexin, were isolated. The CHS cDNA clone (VCHS) showed 87% sequence homology with VvCHS (V.vinifea) and 72.3% identity with VSTSY(V.vinifea). its amino acid sequences were longer than any other CHS genes as 454 residues. Two genes were weakly expressed in white light irradiated cells, but highly induced in UV-B irradiated condition during 32 hours. Interestingly, the STS was quickly and abundantly expressed from 2 hours when supplemented with jasmonic acid (JA) and the maximum expression was observed at 4 hours and then gradually decreased. But, the additional UV-B or white light quickly degraded the STS expression than only JA treated grape suspension cells. The CHS also was rapidly induced with JA and the synergistical effect was observed at the addigional light treatment of UV-B or white light. These results are indicated that CHS and STS have different response mechanisms against the environmental stresses.

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Genetic Relationships of Cattle Breeds Assessed by PCR-RFLP of the Bovine Mitochondrial DNA D-loop Region

  • Yoon, Du Hak;Lee, Hak Kyo;Oh, Sung Jung;Hong, Ki Chang;Jeon, Gwang Joo;Kong, Hong Sik;Lee, Jun Heon
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.10
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    • pp.1368-1374
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    • 2005
  • To investigate the genetic relationships among various cattle breeds, bovine mtDNA D-loop region was used in 411 animals of 18 cattle breeds, including 8 Asian Bos taurus, 7 European Bos taurus, 1 Asian Bos indicus, and 2 African Bos indicus. The size of amplified PCR products from mtDNA D-loop region was 964 bp and the products were digested by 15 different restriction enzymes. Two different band patterns were identified in eight restriction enzymes (BstXI, Hae III, Msp I, Apa I, Taq I, Alu I, BamH I, EcoN I) and the rest of restriction enzymes showed more than 3 different band patterns among which Apo I and MspR9 resulted in 7 different restriction patterns. The genotypes, number of haplotype, effective number of haplotype, and degree of heterozygosity were analyzed. Based on all the PCR-RFLP data, different haplotypes were constructed and analyzed for calculating genetic distances between these breeds using Nei's unbiased method and constructing a phylogenetic tree.

Molecular Identification of Arbuscular Mycorrhizal Fungal Spores Collected in Korea

  • Lee, Jai-Koo;Park, Sang-Hyeon;Eom, Ahn-Heum
    • Mycobiology
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    • v.34 no.1
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    • pp.7-13
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    • 2006
  • Arbuscular mycorrhizas (AM) have mutualistic symbiosis with plants and thus efforts have been placed on application of these symbiotic relationships to agricultural and environmental fields. In this study, AM fungi were collected from 25 sites growing with 16 host plant species in Korea and cultured with Sorghum bicolor in greenhouse condition. AM fungal spores were extracted and identified using both morphological and molecular methods. Using morphological characters, total 15 morpho-speices were identified. DNA was extracted from single spore of AM fungi and a partial region on 18S rDNA was amplified using nested PCR with AM fungal specific primers AML1/AML2. A total of 36 18S rDNA sequences were analyzed for phylogenetic analysis and 15 groups of AM fungi were identified using both morphological and molecular data of spores. Among the species, 4 species, Archaeospora leptoticha, Scutellospora castanea, S. cerradensis, S. weresubiae were described for the first time in Korea and two species in Glomus and a species in Gigaspora were not identified. Morphological and molecular identification of AM fungal spores in this study would help identify AM fungal community colonizing roots.