• Title/Summary/Keyword: 16S-rRNA

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Induction of Microsomal Epoxide Hydrolase, rGSTA2, rGSTA3/5, and rGSTM1 by Disulfiram, but not by Diethyldithiocarbamate, a Reduced Form of Disulfiram

  • Kim, Sang-Geon;Kim, Hye-Jung
    • Toxicological Research
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    • v.13 no.4
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    • pp.339-347
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    • 1997
  • Disulfiram (DSF) and diethyldithiocarbamate (DDC), a reduced form of DSF, protect the liver against toxicant-induced injury through inhibition of cytochrome P450 2E1. The effect of DSF and DDC on the levels of major hepatic microsomal epoxide hydrolase (mEH) and glutathione S-transferase (GST) expression was comparatively studied, given the view that these enzymes are involved in terminal detoxification events for high energy intermediates of xenobiotics. Treatment of rats with a single dose of DSF (20-200 mg/kg, po) resulted in 2- to 15-fold increases in the mEH mRNA level at 24 hr with the ED$_{50}$ value being noted as 60 mg/kg. The mEH mRNA level was elevated ~15-fold at 24 hr after treatment at the dose of 100 mg/kg, whereas the hepatic mRNA level was rather decreased from the maximum at the dose of 200 mg/kg, indicating that DSF might cause cytotoxicity at the dose. In contrast to the effect of DSF, DDC only minimally elevated the mEH mRNA level at the doses employed. DSF moderately increased the major GST mRNA levels in the liver as a function of dose, resulting in rGSTA2, rGSTA3/5 or rGSTM1 mRNA levels being elevated 3- to 4-fold at 24 hr post-treatment, whereas the rGSTM2 mRNA level was not altered. DDC, however, failed to stimulate the mRNA levels for major GST subunits, indicating that the reduced form of DSF was ineffective in stimulating the GST the expression. The effect of other organosulfides including aldrithiol, 2, 2'-dithiobis(benzothiazole) (DTB), tetramethylthiouram disulfide (TMTD) and allyl disulfide (ADS) on the hepatic mEH and GST mRNA expression was assessed in rats in order to further confirm the increase in the gene expression by other disulfides. Treatment of rats with aldrithiol (100 mg/kg, po) resulted in a 16-fold increase in the mEH mRNA level at 24 hr post-treatment. DTB, TMTD and ADS also caused 5-, 9- and 12-fold increases in the rnRNA level, respectively, as compared to control. Thus, all of the disulfides examined were active in stimulating the mEH gene in the liver. The organosulfides significantly increased the rGSTA2, rGSTA3, rGSTA5 and rGSTM1 mRNA levels at 24 hr after administration. In particular, aldrithiol was very efficient in stimulating the rGSTA and rGSTM genes among the disulfides examined. These results provide evidence that DSF and other sulfides effectively stimulate the mEH and major GST gene expression at early times in the liver and that DDC, a reduced form of DSF, was ineffective in stimulating the expression of the genes, supporting the conclusion that reduced form(s) of organosulfur compound(s) might be less effective in inducing the mEH and GST genes through the antioxidant responsive element(s).

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Characterization of the Microbial Diversity in a Korean Solar Saltern by 16S rRNA Gene Analysis

  • Park, Soo-Je;Kang, Cheol-Hee;Rhee, Sung-Keun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1640-1645
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    • 2006
  • We studied the diversity of the halophilic archaea and bacteria in crystallizer ponds of a Korean solar saltern by analyzing 16S rRNA gene libraries. Although diverse halophilic archaeal lineages were detected, the majority (56%) were affiliated with the uncultured and cultured Halorubrum group. Halophilic archaea that have been frequently observed in solar saltern environments previously, such as Halogeometricum, Halococcus, Haloarcula, and Haloferax, were not detected in our samples. The majority of clones (53%) belonged to the Cytophaga-Flavobacterium-Bacteroides and ${\alpha}-,\;{\gamma}-,\;and\;{\delta}-Proteobacteria$ groups, with 47% of the clones being affiliated with ${\gamma}-Proteobacteria$. We also identified new ${\delta}-Proteobacteria$-related bacteria that have not been observed in hypersaline environments previously. Our data show that the diversity of the halophilic archaea and bacteria in our Korean saltern differs from that of solar salterns found in other geographic locations. We also showed by quantitative real-time PCR analysis that bacteria can form a significant component of the microbial community in solar salterns.

A report of 20 unrecorded bacterial species in Korea, isolated from soils of coastal areas in 2022

  • Seung Hyeok Soung;Jaeho Song;Seung Yeol Shin;Song-Ih Han
    • Journal of Species Research
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    • v.12 no.4
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    • pp.267-276
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    • 2023
  • To obtain unrecorded bacterial species in Korea, various soils of coastal areas were collected from the Republic of Korea in 2022. After plating the samples on marine agar and incubating aerobically and anaerobically, approximately 1,700 bacterial strains were isolated and identified using 16S rRNA gene sequences. A total of 20 strains showed ≥98.7% 16S rRNA gene sequence similarity with validly published bacterial species but not reported in Korea, indicating they are unrecorded bacterial species in Korea. The unrecorded bacterial strains belonged to four phyla, six classes, 15 orders, 16 families, and 19 genera which were assigned to Blastomonas and Sphingomonas of the class Alphaproteobacteria; Pseudidiomarina, Kushneria, Salinicola, and Salinisphaera of the class Gammaproteobacteria; Evansella, Virgibacillus, and Paenibacillus of the class Bacilli; Cyclobacterium of the class Cytophagia; Pedobacter of the class Sphingobacteriia; and Demequina, Ornithinimicrobium, Blastococcus, Jatrophihabitans, Kineococcus, Glaciihabitans, Aeromicrobium and Streptomyces of the class Actinomycetes. The details of the 20 unreported species, including Gram reaction, morphology, biochemical characteristics, and phylogenetic position are also provided in the description of the strains.

Molecular Systematics of the Genus Megoura (Hemiptera: Aphididae) Using Mitochondrial and Nuclear DNA Sequences

  • Kim, Hyojoong;Lee, Seunghwan
    • Molecules and Cells
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    • v.25 no.4
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    • pp.510-522
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    • 2008
  • To construct the molecular systematics of the genus Megoura (Hemiptera: Aphididae), DNA based-identification was performed using four mitochondrial and three nuclear DNA regions: partial cytochrome c oxidase I (COI), partial tRNA-leucine + cytochrome c oxidase II (tRNA/COII), cytochrome b (CytB), partial 12S rRNA + tRNA-valine + 16S rRNA (12S/16S), elongation factor-1 alpha ($EF1{\alpha}$), and the internal transcribed spacers 1 and 2 (ITS1, ITS2). Pairwise sequence divergences between taxa were compared, and phylogenetic analyses were performed based on each DNA region separately, and the combined datasets. COI, CytB, $EF1{\alpha}$, ITS1, and ITS2 were relatively effective in determining species and resolving their relationships. By contrast, the sequences of tRNA/COII and 12S/16S were not able to separate the closely related species. CytB and $EF1{\alpha}$ gave better resolution with higher average sequence divergences (4.7% for CytB, 5.2% for $EF1{\alpha}$). The sequence divergence of COI (3.0%) was moderate, and those of the two ITS regions (1.8% for ITS1, 2.0% for ITS2) were very low. Phylogenetic trees were constructed by minimum evolution, maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses. The results indicated that the phylogenetic relationships between Megoura species were associated with their host preferences. Megoura brevipilosa and M. lespedezae living on Lespedeza were closely related, and M. nigra, monophagous on Vicia venosa, was rather different from M. crassicauda, M. litoralis, and M. viciae, which are oligophagous on Lathyrus and Vicia. The three populations of M. crassicauda formed a clade separated from M. litoralis and M. viciae. Nevertheless M. litoralis and M. viciae, which are morphologically similar, were not separated due to negligible sequence divergence. We discuss the phylogenetic relationships of the Megoura, and the usefulness of the seven DNA regions for determining the species level phylogeny of aphids.

Isolation of Bacillus atrophaeus MPL-01 from A Wild Boar and Characterization of Its Antifungal Activity (멧돼지 대장으로부터 Bacillus atrophaeus MPL-01의 분리 및 항진균 활성의 특성)

  • Yun, Sung-Jo;Rho, Jae-Young
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.195-199
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    • 2013
  • A bacterial strain MPL-01 was isolated from the large intestine of a wild boar. The strain was shown to have morphological, physiological and biochemical characteristics, fatty acids composition typical of Bacillus. The 16S rRNA gene sequence showed that the isolate formed distinct phyletic line that was most closely related to this of Bacillus atrophaeus (99.99%). It was proposed that the strain is classified as B. atrophaeus MPL-01. The strain MPL-01 exhibited the strongest antifungal activity against Colletotrichum acutatum, the pathogen of anthracnose of chili peppers. The ethyl acetate extract of culture filtrate possessed not only the antifungal activity but also the bio-surfactant activity. Therefore, the strain MPL-01 could be a useful bacterium in the development of bio-control process against the pathogenic fungi.

Relationship of Oral Bacterial Load Over One Year of Smoking Cessation

  • Kim, Sunghyun;Seo, Min-Seock;Hwang, Soo-Jeong
    • Journal of dental hygiene science
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    • v.19 no.4
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    • pp.213-219
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    • 2019
  • Background: Smoking exerts an adverse effect on the periodontal tissue by reorganizing the ecosystem of oral microorganisms and is considered to be an important factor in the development of periodontal disease. Although cross-sectional studies on smokers and non-smokers have been attempted to investigate the microbial differences in periodontal oral cavity, only few studies have been conducted to investigate the changes in oral microorganisms during smoking cessation. The purpose of this study was to investigate the changes of bacteria in saliva and gingival crevicular fluid (GCF) over a period of one year among 11 smokers trying to quit smoking. Methods: Eleven smokers trying to quit smoking visited the clinic at baseline, two weeks, two months, four months, six months, and 12 months to give saliva and GCF samples. The amounts of 16S rRNA, Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum subsp. nucleatum, Streptococcus mutans, and Streptococcus sobrinus in saliva and GCF were quantified using real-time polymerase chain reaction TaqMan probe assay. The results were analyzed by nonparametric statistical analysis using Friedman test and Spearman correlation coefficient. Results: After cessation of smoking, the amounts of 16S rRNA corresponding to P. gingivalis, F. nucleatum, P. intermedia, and T. denticola in saliva decreased and then again increased significantly. The amount of F. nucleatum 16S rRNA in GCF decreased significantly after smoking cessation. Positive correlations were observed between 16S rRNA and F. nucleatum and between F. nucleatum and T. denticola in saliva and GCF. Conclusion: Even if the number of subjects in this study was small, we suggest that smoking cessation may reduce the total bacterial amount and F. nucleatum in GCF. However, the results regarding changes in the microbial ecosystem due to smoking or smoking cessation were inconsistent. Therefore, further in-depth studies need to be carried out.

Emergence of CTX-M-15 Extended Spectrum β-lactamase and ArmA-Producing Enterobacter cloacae (CTX-M-15형 Extended Spectrum β-lactamase와 ArmA 동시 생성 Enterobacter cloacae의 출현)

  • Sung, Ji-Youn
    • Journal of Digital Convergence
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    • v.13 no.12
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    • pp.313-318
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    • 2015
  • We investigated the prevalence of extended spectrum ${\beta}$-lactamase (ESBL) genes and 16S rRNA methyltransferase genes to study antimicrobial resistance mechanisms of Enterobacter cloacae strains isolated from a university hospital in the Chungcheong province of Korea. Eight of the bacteria strains involved in this study contained CTX-M-15 type ESBL. Among 8 strains harboring the ESBL gene, 3 strains also harbored armA gene. The three isolates showed resistance to antimicrobial agents belonged to third cephalosporin, aminoglycoside, and fluoroquinolones. Furthermore, interspecies plasmid transfer of the antimicrobial resistant genes may induced horizontal spreading of the genes and emergence of multidrug resistant bacteria. Therefore, surveillance for existence of antimicrobial resistance determinants is important to prevent distribution of antimicrobial resistant strains.

Inferring the Molecular Phylogeny of Chroococcalian Strains (Blue-green algae/Cyanophyta) from the Geumgang River, Based on Partial Sequences of 16S rRNA Gene

  • Lee, Wook-Jae;Bae, Kyung-Sook
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.335-339
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    • 2002
  • Partial sequences of 16S rRNA gene of five chroococcalian blue-green algal strains, Aphanothece nidulans KCTC AG10041, Aphanothece naegelii KCTC AG10042, Microcystis aeruginosa KCTC AG10159, Microcystis ichthyoblabe KCTC AG10160, and Microcystis viridis KCTC AG10198, which were isolated from water from the Geumgang River, were determined and were inferred their phylogenetic and taxonomic positions among taxa of order Chroococcales. Most taxa of Chroococcales whose partial 16S rRNA gene sequences were aligned in this study, are clustered with other related taxa. Aphanothece nidulans KCTC AG10041 and Aphanothece naegelii KCTC AG10042 made a cluster with other European species of these genera, which supported 100% of the bootstrap trees with a very high sequence similarity (97.4-99.4%) in this study. Three strains, Microcystis aeruginosa KCTC AG10159, M. ichthyoblabe KCTC AG10160, and M. viridis KCTC AG10198, formed a cluster with other Microcystis spp. supported 100 % of the bootstrap trees with a similarity of 97.0-99.9% except for two strains. However, this phylogentic tree made no resolution among the species of Microcystis spp. The topology of the tree reconfirmed the taxonomic status of three species of Microcystis, identified in this study based on the morphology, as three colonial types of Microcystis aeruginosa com. nov. Otsuka et al. (1999c). The genera of chroococcalian cyanophytes are heterogeneously clustered in these sequence analyses. We suggest that more molecular studies on the genera of Chroococcales with reference strains, widely collected from restricted geographic or environmental ranges, get accurate taxonomic or phylogenetic determinations.

Bacterial Diversity in the Guts of Sea Cucumbers (Apostichopus japonicus) and Shrimps (Litopenaeus vannamei) Investigated with Tag-Encoded 454 Pyrosequencing of 16S rRNA Genes (16S rRNA 유전자의 454 파이로서열 분석을 이용한 해삼(Apostichopus japonicas)과 새우(Litopenaeus vannamei)의 장내 세균의 다양성 연구)

  • Noh, Eun Soo;Kim, Young-Sam;Kim, Dong-Hyun;Kim, Kyoung-Ho
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.237-244
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    • 2013
  • Bacterial diversities in the guts of sea cucumbers (Apostichopus japonicus) and shrimps (Litopenaeus vannamei) were investigated using barcoded or tag-encoded 454 pyrosequencing of 16S rRNA genes. In sea cucumbers, most of sequences were related to two genera, the genus Propionigenium in the phylum Fusobacteria and an unclassified genus in the family Flavobacteriaceae of phylum Bacteroidetes. Shrimps showed various kinds of genera including Lactococcus, Leuconostoc, Prochlorococcus, and Vibrio as well as the unclassified genera in the families, Flavobacteriaceae, Rhodobacteraceae, Desulfobulbaceae, and Helicobacteraceae and in the order Mycoplasmatales. Unclassified genera containing environmental sequences only are more than half of genera from sea cucumbers and shrimps. Sea cucumbers and shrimps could be unexplored sources of novel microbes and the bacterial diversity of them was revealed by high throughput 454 pyrosequencing.

Selection of Stable Reference Genes for Real-Time Quantitative PCR Analysis in Edwardsiella tarda

  • Sun, Zhongyang;Deng, Jia;Wu, Haizhen;Wang, Qiyao;Zhang, Yuanxing
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.112-121
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    • 2017
  • Edwardsiella tarda is a gram-negative pathogenic bacterium in aquaculture that can cause hemorrhagic septicemia in fish. Many secreted proteins have already been identified as virulent factors of E. tarda. Moreover, since virulent phenotypes are based on the expression regulation of virulent genes, understanding the expression profile of virulent genes is important. A quantitative RT-PCR is one of the preferred methods for determining different gene expressions. However, this requires the selection of a stable reference gene in E. tarda, which has not yet been systematically studied. Accordingly, this study evaluated nine candidate reference genes (recA, uup, rpoB, rho, topA, gyrA, groEL, rpoD, and 16S rRNA) using the Excel-based programs BestKeeper, GeNorm, and NormFinder under different culture conditions. The results showed that 16S rRNA was more stable than the other genes at different culture growth phases. However, at the same culture time, topA was identified as the reference gene under the conditions of different strains, different culture media, and infection, whereas gyrA was identified under the condition of different temperatures. Thus, in experiments, the expression of gapA and fbaA in E. tarda was analyzed by RT-qPCR using 16S rRNA, recA, and uup as the reference genes. The results showed that 16S rRNA was the most suitable reference gene in this analysis, and that using unsuitable reference genes resulted in inaccurate results.