• Title/Summary/Keyword: Bioassays

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Human Lung Insults due Air Pollutant -A Review for Priority Setting in the Research- (대기오염에 의한 폐장조직 손상 -연구방향의 설정을 위한 논의-)

  • 김건열;백도명
    • Journal of environmental and Sanitary engineering
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    • v.7 no.2
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    • pp.95-110
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    • 1992
  • Much progress has been made in understanding the subcellular events of the human lung injuries after acute exposure to environmental air pollutants. Host of those events represent oxidative damages mediated by reactive oxygen species such as superoxide, hydrogen peroxide, and the hydroxy, free radical. Recently, nitric oxide (NO) was found to be endogenously produced by endothelial cells and cells of the reticulo-endothelial system as endothelialderived relaxation factor (EDRF) which is a vasoactive and neurotransmitter substance. Together with superoxide, NO can form another strong oxidant, peroxonitrite. The relative importance of exogenous sources of $N0/N0_2$ and endogenous production of NO by the EDRF producing enzymes in the oxidative stresses to the heman lung has to be elucidated. The exact events leading to chronic irreversible damage are still yet to be known. From chronic exposure to oxidant gases, progressive epithelial and interstitial damages develop. Type I epithelial cells become thicker and cover a smaller average alveolar surface area while thee II cells proliferate instead. Under acute damages, the extent of loss of the alveolar epithelial cell lining, especially type II cells appears to be a good predictor of the ensuing irreversible damage to alveolar compartment. Interstitial matrix undergo remodeling during chronic exposure with increased collagen fibers and interstitial fibroblasts. However, Inany of these changes can be reversed after cessation of exposure. Among chronic lung injuries, genetic damages and repair responses received particular attention in view of the known increased lung cancer risks from exposure to several air pollutants. Heavy metals from foundry emission, automobile traffics, and total suspended particulate, especially polycystic aromatic hydrocarbons have been positively linked with the development of lung cancer. Asbestos in another air pollutant with known risk of lung cancer and mesothelioma, but asbestos fibers are nonauthentic in most bioassays. Studies using the electron spin resonance spin trapping method show that the presence of iron in asbestos accelerates the production of the hydroxy, radical in vitro. Interactions of these reactive oxygen species with particular cellular components and disruption of cell defense mechanisms still await further studies to elucidate the carcinogenic potential of asbestos fibers of different size and chemical composition. The distribution of inhaled pollutants and the magnitude of their eventual effects on the respiratory tract are determined by pollutant-independent physical factors such as anatomy of the respiratory tract and level and pattern of breathing, as well as by pollutant-specific phyco-chemical factors such as the reactivity, solubility, and diffusivity of the foreign gas in mucus, blood and tissue. Many of these individual factors determining dose can be quantified in vitro. However, mathematical models based on these factors should be validated for its integrity by using data from intact human lungs.

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A Study on the Degradation of Parathion and Reduction of Acute Toxicity in Solar $TiO_2$ Photocatalysis (태양광 $TiO_2$ 광촉매를 이용한 Parathion의 분해와 독성저감에 관한 연구)

  • Kim, Tak-Soo;Kim, Jung-Kon;Choi, Kyung-Ho;Zoh, Kyung-Duk
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.8
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    • pp.822-828
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    • 2005
  • The photocatalytic degradation of methyl parathion was carried out using a circulating $TiO_2$/solar system. Under the photocatalytic condition, parathion was more effectively degraded than by the photolysis or $TiO_2$ only condition. The parathion degradation followed pseudo first-order kinetics. With photocatalysis, 10 mg/L parathion was completely degraded within 90 min with a TOC decrease exceeding 63% after 150 min. The nitrogen from parathion was recovered mainly as ${NO_2}^-$, ${NO_3}^-$, and ${NH_4}^+$, 80% of sulfur as ${SO_4}^{2-}$, and less than 5% of phosphorus as ${PO_4}^{3-}$ during photocatalysis. The organic intermediates 4-nitrophenol and paraoxon were also identified, and these were further degraded. Two different bioassays using V. fischeri and D. magna were employed to measure the toxicity reduction in the solutions treated by both photocatalysis and photolysis. Relative toxicity was reduced almost completely after 150 min in both organisms under the photocatalysis, whereas in photolysis, 76 and 57% reduction was achieved for V. fischeri and D. magna, respectively. The acute toxicity reduction pattern corresponded with the decrease in parathion and TOC concentrations.

Root Colonization and Quorum Sensing of the Antagonistic Bacterium Pseudomonas fluorescens 2112 involved in the Red-pepper Rhizosphere (생물방제균 Pseudomonas fluorescens 2112의 고추 근권정착능과 Quorum-sensing 기능)

  • Jung, Byung-Kwon;Kim, Yo-Hwan;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.105-111
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    • 2013
  • Biofilm formation of multifunctional plant growth promoting rhizobacterium (PGPR), Pseudomonas fluorescens 2112 is necessary for P. fluorescens 2112 to have a positive impact on the rhizosphere of red-pepper. This study investigated whether signal molecules of the quorum sensing AHLs are produced in order to confirm biofilm formative ability. Through the use of Petri dish bioassays a blue circle formed evidence of AHLs. It was confirmed that P. fluorescens 2112 produced six-carbon-chain-long AHLs by TLC bioassay. The bacterial density of P. fluorescens 2112 on the top and bottom of pepper plant roots was estimated as $3{\times}10^5$ and $8{\times}10^3$ CFU/g root, respectively. P. fluorescens 2112 exist more with high-density of $3.5{\times}10^6$ CFU/g soil at a depth of 1 cm but at a low-density of $1.1{\times}10$ CFU/g soil at a depth of 5 cm, from the surface of rhizosphere soil. In addition, biofilm formation of P. fluorescens 2112 on the epidermises and the tips of the red-pepper roots were confirmed visually by SEM. Thus, the production of AHLs by P. fluorescens 2112 brings about quorum sensing signaling and the formation of biofilm on the roots which has a positive effect on economically important crops such as red-pepper by additionally producing a variety of antifungal substances and auxin.

Application and utilization of marker assisted selection for biotic stress resistance in hybrid rice (Oryza sativa L.)

  • Song, Jae-Young;Ouk, Sothea;Nogoy, Franz Marielle;Nino, Marjohn C.;Kwon, Soon Wook;Ha, Woongoo;Kang, Kwon-Kyoo;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.317-331
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    • 2016
  • Development of disease resistant plant is one of the important objectives in rice breeding programs because biotic stresses can adversely affect rice growth and yield losses. This study was conducted to identify lines with multiple-resistance genes to biotic stress among 173 hybrid rice breeding lines and germplasms using DNA-based markers. Our results showed that one hybrid rice line [IR98161-2-1-1-k1-3 (IR86409-3-1-1-1-1-1/IRBB66)] possessed 5 bacterial blight resistance genes (Xa4, xa5, Xa7, Xa13 and Xa21) while two hybrid rice lines [IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66) and 7292s (IR75589-31-27-8-33S(S1)/IR102758B)] possessed 3 bacterial blight resistance genes (Xa4, Xa7 and Xa21, and Xa3, Xa4 and xa5). Molecular survey on rice blast disease revealed that most of these lines had two different resistant genes. Only 11 lines possessed Pib, Pi-5, and Pi-ta. In addition, we further surveyed the distribution of insect resistant genes, such as Bph1, Bph18(t), and Wbph. Three hybrid breeding lines [IR98161-2-1-1-k1-3 (IR86409-3-1-1-1-1-1/IRBB66), IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66), and 7292s (IR75589-31-27-8-33S(S1) /IR102758B)] contained all three resistance genes. Finally, we obtained four hybrid rice breeding lines and germplasms [IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66), Damm-Noeub Khmau, 7290s, and 7292s (IR75589-31-27-8-33S(S1)/IR102758B)] possessing six-gene combination. They are expected to provide higher level of multiple resistance to biotic stress. This study is important for genotyping hybrid rice with resistance to diverse diseases and pests. Results obtained in this study suggest that identification of pyramided resistance genes is very important for screening hybrid rice breeding lines and germplasms accurately for disease and pest resistance. We will expand their cultivation safely through bioassays against diseases, pests, and disaster in its main export countries.

Improved Soil Application Bioassay for Efficient Development of Natural Pre-emergence Herbicides (토양처리용 천연물제초제 개발을 위한 생물검정법 개선)

  • Kim, Jae-Deog;Hwang, Hyun-Jin;Seo, Bo-Ram;Choi, Jung-Sup;Kim, Jin-Seog
    • Korean Journal of Weed Science
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    • v.31 no.3
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    • pp.229-239
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    • 2011
  • This study was carried out to establish an improved bioassay system on the side of practicality, pre-emergence bioassay which is more effective in developing soil application natural herbicides. A miniaturized method which have a 50 cm2 of soil surface area and was efficient by 7 times compared to the existing soil application assay ($350cm^2$ of soil surface area) was established, in which four weed species (Echinochloa crus-galli, Digitaria sanguinalis, Aeschynomene indica, and Abutilon theophrasti) were planted and grown in greenhouse. This would be applicable when the amount of screening compound is much more than 50 mg. The initial application rate was desirable at $10,000{\mu}g\;mL^{-1}$. On the other hand, the 6 well plate assay which has 4 weed species in each well containing upland soil and could be conducted in growth chamber, was established. This assay was resulted in minimizing in level of 1/14 test volume and 1/14 amounts of test compound to the conventional method that has been used for screening of synthetic compounds in KRICT, and applicable for the small amount of test compound (less than 10 mg). Therefore, the improved bioassays established in this study would be helpful for a rapid and efficient development of soil application natural herbicides.

Identification and Biological Activity of Two New Phytotoxins Isolated from Botrytis cinerea (Botrytis cinerea로부터 분리한 두 개의 새로운 phytotoxin의 구조 결정 및 생물활성)

  • Kim, Geum-Jung;Yoon, Mi-Young;Kim, Heung-Tae;Choi, Gyung-Ja;Jang, Kyoung-Soo;Choi, Yong-Ho;Park, Myung-Soo;Cha, Byeong-Jin;Kim, Jin-Cheol
    • Research in Plant Disease
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    • v.15 no.2
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    • pp.112-119
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    • 2009
  • We discovered two novel phytotoxins produced by the pathogenic fungus, Botrytis cinerea. Among the twenty-five B. cinerea isolates, which were obtained from various host plants in 1994 and 1996, twenty-two showed strong or moderate pathogenicity on five plants such as cucumber, tomato, red pepper, tobacco and Chinese cabbage. The culture filtrate of the B. cinerea 2-16 strain showed the most potent phytotoxic activity in a tobacco leaf-wounding assay. Two novel phytotoxins were isolated from the liquid cultures of B. cinerea 2-16 by ethyl acetate extraction, flash silica gel column chromatography, silica gel column chromatography, Sephadex LH-20 column chromatography, preparative TLC and subsequently preparative HPLC. Their chemical structures were determined to be 3-O-acetyl botcinol and 3-O-acetyl botcinolide, respectively, by mass and NMR spectral analyses. These two phytotoxins caused leaf necrosis in a leaf-wounding bioassay, and significant electrolyte leakage from leaf tissues of tobacco. In the two bioassays tested, 3-O-acetyl botcinol exhibited stronger phytotoxic activity than 3-O-acetyl botcinolide. This is the first report on the production of both 3-O-acetyl botcinol and 3-O-acetyl botcinolide from B. cinerea.

Conditions for Stable light Production of Recombinant Escherichia coli Containing Lux Operon and Sensitivity to Toxic Chemicals (Lux operon을 함유한 유전자 재조합 Escherichia coli의 발광 안정화 조건 및 독성물질에 대한 민감성)

  • 배희경;이상민;정윤철;송방호;신평균
    • KSBB Journal
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    • v.17 no.6
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    • pp.571-576
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    • 2002
  • Recombinant E. coli DH5 ${\alpha}$/pSB311 was made by cloning the genes encoding bacterial luciferase and aldehyde substrate proteins from Photohabdus luminescense, to complement defects of Lumistox, which is normally used in bioassays to monitor toxic substances in water environmental systems. The conditions for stable light production by the recombinant strains were investigated with respect to cell growth stage, cell number, and buffer conditions. The optimum growth stage was a middle-exponential stage with an OD$_{660nm}$ value of 0.6-0.7. ADout 10$^{6}$-10$^{7}$ cells per test tube was optimum for stable light emission. The effect of buffer was not significant if an optimum viable cell number was maintained. The bioluminescence of the recombinant E. coli harboring the lux operon of Photohabdus luminescense was not affected by temperature, while the bioluminescence of Lumistox was temperature sensitive. The recombinant E. coli was more sensitive to heavy metals (Cd, Cu, Hg, Zn) than Lumistox, because it does not require high concentrations of NaCl in the buffer.

Gut Bacterial Diversity of Insecticide-Susceptible and -Resistant Nymphs of the Brown Planthopper Nilaparvata lugens Stål (Hemiptera: Delphacidae) and Elucidation of Their Putative Functional Roles

  • Malathi, Vijayakumar M.;More, Ravi P.;Anandham, Rangasamy;Gracy, Gandhi R.;Mohan, Muthugounder;Venkatesan, Thiruvengadam;Samaddar, Sandipan;Jalali, Sushil Kumar;Sa, Tongmin
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.976-986
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    • 2018
  • Knowledge about the gut bacterial communities associated with insects is essential to understand their roles in the physiology of the host. In the present study, the gut bacterial communities of a laboratory-reared insecticide-susceptible (IS), and a field-collected insecticide-resistant (IR) population of a major rice pest, the brown planthopper Nilaparvata lugens, were evaluated. The deep-sequencing analysis of the V3 hypervariable region of the 16S rRNA gene was performed using Illumina and the sequence data were processed using QIIME. The toxicological bioassays showed that compared with the IS population, IR population exhibited 7.9-, 6.7-, 14.8-, and 18.7-fold resistance to acephate, imidacloprid, thiamethoxam, and buprofezin, respectively. The analysis of the alpha diversity indicated a higher bacterial diversity and richness associated with the IR population. The dominant phylum in the IS population was Proteobacteria (99.86%), whereas the IR population consisted of Firmicutes (46.06%), followed by Bacteroidetes (30.8%) and Proteobacteria (15.49%). Morganella, Weissella, and Enterococcus were among the genera shared between the two populations and might form the core bacteria associated with N. lugens. The taxonomic-to-phenotypic mapping revealed the presence of ammonia oxidizers, nitrogen fixers, sulfur oxidizers and reducers, xylan degraders, and aromatic hydrocarbon degraders in the metagenome of N. lugens. Interestingly, the IR population was found to be enriched with bacteria involved in detoxification functions. The results obtained in this study provide a basis for future studies elucidating the roles of the gut bacteria in the insecticide resistance-associated symbiotic relationship and on the design of novel strategies for the management of N. lugens.

Antioxidant, Antimicrobial and Anti-inflammatory Activities of Essential Oil from Erigeron annuus L. Flower (개망초꽃 에센셜 오일의 항산화, 항균 및 항염 활성)

  • Yi, Mi-Ran;Jeon, Ah-Lim;Kang, Chang-Hee;Bu, Hee-Jung
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.4
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    • pp.717-725
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    • 2016
  • This study was designed to examine the in vitro antioxidant, antimicrobial and anti-inflammation effects of essential oils of Erigeron annuus L. Flower. Erigeron annuus L. essential oils were obtained by solvent extraction. Antioxidative ability was evaluated by bioassays using ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid diammonium salt) radical scavenging effect and 2, 2-diphenyl-1-1-picrydrazyl (DPPH) free radical scavenging activity. Erigeron annuus L. essential oil exhibited free radical scavenging activity on ABTS and DPPH 98.6%, 48.3% respectively, at a concentration of $500{\mu}g/ml$. Antimicrobial activity of essential oils of Erigeron annuus L. were tested against Staphylococcus aureus (S. aureus), Propionibacterium acnes (P. acne) and Escherichia coli (E. coli) by paper disc method, MIC and MBC. Erigeron annuus L. essential oil showed excellent antibacterial activities against S. aureus with MIC and MBC values of 0.31 mg/mL. The clear zone, indicating antimicrobial activity against P. acnes, was 14 mm, MIC and MBC values 0.31 mg/mL, 0.63 mg/mL, respectively. For the anti-inflammatory activity in RAW 264.7 cell, the Erigeron annuus L. essential oils inhibited not only NO production but also the expression of pro-inflammatory cytokines such as, TNF-${\alpha}$, IL-6 in a dose-dependent manner. These results suggested that Erigeron annuus L. essential oils has considerable potential as a cosmetic ingredient with antioxidative, antimicrobial and anti-inflammation effects.

Antioxidation and Anti-inflammation Activity of Isodon inflexus (Thunb.) Kudo Extract and Its Isolated Substance (산박하 추출물과 분리물질의 항산화 및 항염증 활성)

  • Bu, Hee-Jung;Lee, Sunjoo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.3
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    • pp.257-268
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    • 2016
  • The functions of anti-oxidation and anti-inflammation were investigated with the crude 80% methanol extract, subfractions and henryin isolated from Isodon inflexus (Thunb.) Kudo (I. inflexus (Thunb.) Kudo). Antioxidative ability was evaluated by bioassays using 2, 2-diphenyl-1-1-picrydrazyl (DPPH) free radical scavenging activity, xanthine oxidase inhibition, and superoxide radical scavenging effects. Ethyl acetate and butanol fractions exhibited free radical scavenging activity on superoxide with $IC_{50}$ values of $0.9{\mu}g/mL$, $0.2{\mu}g/mL$, respectively, which were stronger activity than that of allopurinol ($2.2{\mu}g/mL$) as reference. For the inhibition of anti-inflammatory activity in RAW 264.7 cell, the ethyl acetate fraction showed strong inhibition activity NO production, and henryin isolated from its subfraction reduced the activity in a dose-dependent manner. Ethyl acetate fraction and henryin suppressed not only mRNA expression of iNOS and COX-2, but also the mRNA expression of pre-inflammatory cytokines such as, TNF-${\alpha}$, 1L-$1{\beta}$, IL-6, in a dose-dependent manner. These results suggested that ethyl acetate fraction of I. inflexus (Thunb.) Kudo has considerable potential as a cosmetics ingredient with an antioxidant and anti-inflammatory effects and henryin can be applied as an functional reference.