• Title/Summary/Keyword: GC analysis

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Analysis of in planta Expressed Orphan Genes in the Rice Blast Fungus Magnaporthe oryzae

  • Sadat, Md. Abu;Jeon, Junhyun;Mir, Albely Afifa;Kim, Seongbeom;Choi, Jaeyoung;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.30 no.4
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    • pp.367-374
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    • 2014
  • Genomes contain a large number of unique genes which have not been found in other species. Although the origin of such "orphan" genes remains unclear, they are thought to be involved in species-specific adaptive processes. Here, we analyzed seven orphan genes (MoSPC1 to MoSPC7) prioritized based on in planta expressed sequence tag data in the rice blast fungus, Magnaporthe oryzae. Expression analysis using qRT-PCR confirmed the expression of four genes (MoSPC1, MoSPC2, MoSPC3 and MoSPC7) during plant infection. However, individual deletion mutants of these four genes did not differ from the wild-type strain for all phenotypes examined, including pathogenicity. The length, GC contents, codon adaptation index and expression during mycelial growth of the four genes suggest that these genes formed during the evolutionary history of M. oryzae. Synteny analyses using closely related fungal species corroborated the notion that these genes evolved de novo in the M. oryzae genome. In this report, we discuss our inability to detect phenotypic changes in the four deletion mutants. Based on these results, the four orphan genes may be products of de novo gene birth processes, and their adaptive potential is in the course of being tested for retention or extinction through natural selection.

Analysis of Agrochemical Residues in Tobacco Using Solid Phase Microextraction-Gas Chromatography with Different Mass Spectrometric Techniques

  • Lee, Jeong-Min;Jang, Gi-Chul;Kim, Hyo-Keun;Hwang, Geon-Joong
    • Journal of the Korean Society of Tobacco Science
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    • v.30 no.2
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    • pp.117-124
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    • 2008
  • A solid phase microextraction (SPME) method in combination with gas chromatography/mass spectrometric techniques was used for the extraction and quantification of 12 selected agrochemical residues in tobacco. The parameters such as the type of SPME fiber, adsorption/desorption time and the extraction temperature affecting the precision and accuracy of the SPME method were investigated and optimized. Among three types of fibers investigated, polyacrylate (PA), polydimethylsiloxane (PDMS) and polydimethylsiloxane-divinylbenzene (PDMS-DVB), PDMS fiber was selected for the extractions of the agrochemicals. The SPME device was automated and on-line coupled to a gas chromatograph with a mass spectrometer. Mass spectrometry (MS) was used and two different instruments, a quadrupole MS and triple quadrupole MS-MS mode, were compared. The performances of the two GC-MS instruments were comparable in terms of linearity (in the range of 0.01$\sim$0.5 $\mu$g/mL) and sensitivity (limits of detection were in the low ng/mL range). The triple quadrupole MS-MS instrument gave better precision than that of quadrupole MS system, but generally the relative standard deviations for replicates were acceptable for both instruments (< 15%). The LODs was fully satisfied the requirements of the CORESTA GRL. Recoveries of 12 selected agrochemicals in tobacco yielded more than 80% and reproducibility was found to be better than 10% RSD so that SPME procedure could be applied to the quantitative analysis of agrochemical residues in tobacco.

Anti-inflammatory Activity of Essential Oil Extracted from Chamaecyparis obtusa (Sieb. et Zucc.) Wood (편백 목부 정유의 항염증 효과 평가)

  • Yang, Jiyoon;Ahn, Changhwan;Jeung, Eui-Bae;Choi, Won-Sil;Kim, Jae-Woo;Park, Mi-Jin
    • Korean Journal of Pharmacognosy
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    • v.48 no.1
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    • pp.18-24
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    • 2017
  • This study was conducted to determine the anti-inflammatory effect of essential oil extracted from the wood of Chamaecyparis obtusa Sieb. et Zucc. Endl. (Cupressaceae). The essential oil was extracted from the wood of C. obtusa by hydrodistillation method, and conducted the analysis on the chemical composition of the extracted C. obtusa wood oil through GC-MS. The major constituents of the oil were found to be: ${\alpha}-pinene$ (11.4%), cadinene (5.4%), ${\delta}-cadiene$ (9.0%), ${\tau}-muurolol$ (22.2%), ${\alpha}-cadinol$ (20.8%) etc. We attempted to identify the anti-inflammatory activities of the oil when it is injected in the lipopolysaccharide (LPS)-stimulated RBL-2H3 cells, along with its effects on the secretion of interleukin-4 (IL-4), interleukin-13 (IL-13), ${\beta}-hexosaminidase$. According to the cell viability analysis conducted by MTT assay, the oil in $10^{-7}{\sim}10^{-5}%$concentration showed no effect on the cell viability. After RBL-2H3 cells treated by LPS stimulation were exposed to $10^{-7}%$ concentration of C. obtusa wood oil, the expression levels of IL-4, IL-13 within the cell were observed to remarkably decrease. Also, it was attenuated the release of ${\beta}-hexosaminidase$ from mast cells to a significantly meaningful level. These results suggest that C. obtusa wood oil exerts the anti-inflammatory effect, by regulating the expression of inflammatory cytokines, which is a valuable feature to be highly utilized as the functional materials in the future.

Comparison of the accuracy of digital models made from white light scanner by scanning method (스캐닝 방법에 따른 백색광 스캐너 기반으로 채득된 디지털 모형의 정확성 비교)

  • Kim, Ki-Baek;Lee, Gyeong-Tak;Kim, Jae-Hong
    • Journal of Korean society of Dental Hygiene
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    • v.12 no.6
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    • pp.1082-1089
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    • 2012
  • Objectives : The aim of this study was to determine the accuracy of digitized stone models, impression materials compared to the master model and the reliability of the computer aided analysis. Methods : A master model(500B-1, Nissin dental product, Japan) with the prepared lower full arch tooth was used. Ten vinyl polysiloxane impressions(Examix$^{(R)}$, GC Industrial Corp, Japan) of master model were taken and type IV stone(aesthetic-base gold$^{(R)}$, Dentona, Germany) were poured in stone models. The linear distance between the reference points were measured and analyzed on the Delcam Copycad$^{(R)}$(Delcam plc, UK). The t-student test for paired samples was used for statistical analysis. Results : The mean differences to master model for stone model and impression material were 0.11~0.19mm, and 0.19~0.29mm, respectively. There were statistical differences in dimensional accuracy for full arch impression between master model and stone model/impressions(p<.05). Conclusions : Two different scanning methods showed clinically acceptable accuracy of full arch digital impression produced by them. These results will have to be confirmed in further clinical studies.

Development of Emission Factors for Greenhouse Gas CO2) from Anthracite Fired Power Plants in Korea (무연탄 화력발전소의 이산화탄소 배출계수 개발)

  • Jeon, Eui-Chan;Myeong, Soo-Jeong;Jeong, Jae-Hak;Lee, Sung-Ho;Sa, Jae-Whan;Roh, Gi-Hwan;Kim, Ki-Hyun;Bae, Wi-Sup
    • Journal of Korean Society for Atmospheric Environment
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    • v.23 no.4
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    • pp.440-448
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    • 2007
  • Although the anthracite power plant is an important source of greenhouse gas, research on this type of power plant has not been conducted much. The present study investigated the entire anthracite power plants in Korea and analyzed the emitted gas in connection with GC/FD and a methanizer in order to develop $CO_2$ emission factors. The study also sampled the anthracite to analyze the amount of carbon and hydrogen using an element analyzer, and to measure the calorie using an automatic calorie analyzer. The emission factors computed through the fuel analysis was 30.45 kg/GJ and that computed through the $CO_2$ gas analysis was 26.48 kg/GJ. The former is approximately about 15% higher than the latter. When compared the carbon content factors of anthracite with that of bituminous coal, the value of anthracite was 24% higher Compared with IPCC values, the emission factors by the fuel was 14% higher, and that by the emitted $CO_2$ gas was about 1.2% lower. More research is needed on our own emission factors of various energy-consuming facilities in order to stand on a higher position in international negotiations regarding the treaties on climate changes.

Kinetic behavior of sophoricoside by gas chromatography/mass spectrometry in rats

  • Jeon, Hee-Kyung;Park, Hae-Yeon;Kim, Youn-Jung;Kim, Youngsoo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.189-189
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    • 2003
  • Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). To develope as novel anti-allergic drug, kinetic study was performed in rats. Serum concentration of sophoricoside was measured by gas chromatography-mass spectrometry (GC/MS) in male Sprague-Dawley rat (250${\pm}$10g, n=5) after oral administration of sophoricoside (100mg/kg). The recovery of sophoricoside after extraction and concentration was above 95 % from rat serum. Between-day precision(relative standard deviation 2.2-2.8%) and within-day precision(2.0-12.1%) were determined from replicate analysis of a spiked control and incurred serum sample. The detection limits of sophoricoside in this serum was approximately 0.1 ng/mL. The Pharmacokinetic parameters were derived from the noncompartmental analysis. The C$\_$max/(3.56${\pm}$0.34 $\mu\textrm{g}$/mL) value for sophoricoside in male rat was observed at 7.6 h. The elimination half-life(t$\_$1/2/) of sophoricoside was approximately 4.47 h, the mean residence time (MRT) averaged 10.75 h, the total body clearance (Cl) averaged 0.0042 mL/min/kg. and the area under the serum concentration-time curve (AUC$\_$0-$\infty$/) was 24.93 $\mu\textrm{g}$$.$hr/mL.

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Effectiveness of Bioremediation on Oil-Contaminated Sand in Intertidal Zone

  • Oh, Young-Sook;Sim, Doo-Suep;Kim, Sang-Jin
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.437-443
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    • 2003
  • Bioremediation technologies were applied to experimental microcosms, simulating an oil spill in a lower intertidal area. Three treatments (oil only, oil plus nutrients, and oil plus nutrients and microbial inocula) were applied, and each microcosm was repeatedly filled and eluted with seawater every 12 h to simulate tidal cycles. To minimize washing-out of the inoculum by the tidal cycles, microbial cells were primarily immobilized on diatomaceous earth before they were applied to the oiled sand. Oil degradation was monitored by gravimetric measurements, thin layer chromatography/flame ionization detector (TLC/FID) analysis, and gas chromatography (GC) analysis, and the loss of oil content was normalized to sand mass or nor-hopane. When the data were normalized to sand mass, no consistent differences were detected between nutrient-amended and nutrient/inoculum-amended microcosms, although both differed from the oil-only microcosm in respect of oil removal rate by a factor of 4 to 14. However, the data relative to nor-hopane showed a significant treatment difference between the nutrient-amended and nutrient/inoculum-treated microcosms, especially in the early phase of the treatment. The accelerating effect of inoculum treatment has hardly been reported in studies of oil bioremediation in the Tower intertidal area. The inoculum immobilized on diatomaceous earth seemed to be a very effective formulation for retaining microbial cells in association with the sand. Results of this study also suggest that interpretation of the effectiveness of bioremediation could be dependent on the selection of monitoring methods, and consequently the application of various analytical methods in combination could be a solution to overcome the limitations of oil bioremediation monitoring.

An Assessment of Genotoxicity on Organic Solvent Workers by $^{32}$P-postlabelling Method ($^{32}$P-postlabelling법을 이용한 유기용제 작업장 근로자의 유전독성 평가)

  • 홍대용;김장락;이장호;문중갑;이한우;김동일;박성학;정주화;이홍근
    • Environmental Analysis Health and Toxicology
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    • v.9 no.1_2
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    • pp.37-51
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    • 1994
  • To evaluate the genotoxicities of workers exposed to glue and glue cleaning solution, ambient air monitoring of working place, animal study and human monitoring were carried out. By GC-MS analysis, air samples collected from shoesmaking plant were found to be toluene, xylene, cyclohexane, n-hexane, methyl ethyl ketone, trichloroethylene, butylacetate, isopropyl alcohol. Glue and glue cleaning solution from shoesmaking plant were applicated topically to the CD-1 mice. DNA was isolated from skin 24 hr following the application and analysed for DNA-adducts using the nuclease $P_1$version of $^{32}$P-postlabelling assay. RAL (Relative Adduct Labelling, adducts$10^8$ nucleotides) was significantly increased in a dose-dependent manner in the glue cleaning solution treated mice skin. Peripheral blood DNA-adducts of workers exposed to glue and glue cleaning solution were also analysed by the same method, but there were not significant differences in the peripheral blood DNA-adducts level between exposed and control workers. In addition, glue cleaning solution from shoes factory was evaluated for mutagenicity in the Salmonella plate incorporation assay using strains TA 100 and TA 1535 in the presence and absence of Arochlor 1254-induced rat liver S$_{9}$. There was evident mutagenicity for cleaning solution in TA 100 regardless of $S_9$, but TA 1535 showed positive only in the absence of $S_9$when predicted by Stead model of mutagenicity prediction (p=0.0000). The urine concentrates from workers and controls were also assayed for mutagenicity towards strain TA 100 of Salmonella typhimurium in the presence of $S_9$ using Kado's microsuspension assay, but their mutagenic activities were not found to be significant. These data suggest that shoesmaking workers are exposed to genotoxic compounds and need to be monitored by testing the mutagenicity of human urines. However, $^{32}$P-postlabelling application requires further validation for the routine monitoring of human exposure.osure.

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Genetic Relationships of Four Korean Oysters Based on RAPD and Nuclear rDNA ITS Sequence Analyses

  • Kim, Woo-Jin;Lee, Jeong-Ho;Kim, Kyung-Kil;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee-Jeong;Jung, Hyung-Taek
    • The Korean Journal of Malacology
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    • v.25 no.1
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    • pp.41-49
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    • 2009
  • Random amplified polymorphic DNA (RAPD) marker and sequence analyses of the internal transcribed spacer (ITS) region of ribosomal DNA were used to assess phylogenetic relationships of four Korean oyster species. The average number of species-specific markers identified from five universal rice primers (URPs) by RAPD-PCR was 1.8 for Crassostrea gigas, 3.2 for C. nippona, 3.6 for C. ariakensis, and 4.6 for Ostrea denselamellosa. The length of the ITS (ITS1-5.8S-ITS2) region ranged from 1,001 to 1,206 bp (ITS1, 426-518 bp; 5.8S, 157 bp; and ITS2, 418-536 bp), while the GC content ranged from 55.5-61.1% (ITS1, 56.8-61.8%; 5.8S, 56-57.3%; and ITS2, 54.1-62.2%). A phylogenetic analysis of the oysters based on our RAPD, ITS1, and ITS2 sequence data revealed a close relationship between C. gigas and C. nippona and a distant relationship between the genera Crassostrea and Ostrea. Our results indicated that RAPD and ITS sequence analysis was a useful tool for the elucidation of phylogenetic relationships and for the selection of species-specific markers in Korean oysters.

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Volatile Sulfur Compounds, Proximate Components, Minerals, Vitamin C Content and Sensory Characteristics of the Juices of Kale and Broccoli Leaves (케일 및 브로콜리잎즙의 함황 향기성분, 일반성분, 무기질, Vitamin C 함량 및 관능적 특성)

  • 김미리;김진희;위대성;나종현;석대은
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.6
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    • pp.1201-1207
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    • 1999
  • To utilize the leaves of broccoli, the shape of which was similar with that of kale leaves, as a vegetable juice, the extracts from two types of vegetable leaves were subjected to the analyses of proximate com ponents, volatile sulfur compounds, vitamin C and minerals. The sensory evaluation of the juices, prepared from kale and broccoli leaves, were performed by duo trio test and scoring test. Among eight varieties of broccoli, four varieties('Pilgrim', 'Greenbelt', 'Salinas' and 'Shasta')were not significantly different from kale(p<0.05), based on the duo trio sensory test. Sulfur and nitrogen compounds present in the dichloromethane extracts of kale and broccoli leaves 'Pilgrim' were identified as 3 butenyl, allyl, butyl and 4 methylsulfinylbutyl isothiocyanate(sulforaphane), 4,5 epithiovalero, 4,5 epithio 3 hydroxyvalero, benzenepropane, 3 hydroxy 4 propene, 4 methylthio butane, 3 hydroxy 3 phenylpropane, 5 methy lsulfinylpentane, 4 methoxyphenyl 3 hydroxypropane nitrile and dimethyl trisulfide by GC/MSD analysis. Proximate components were observed to be relatively similar between kale and broccoli leaves. Vitamin C and sugar content were higher in broccoli leaves(125∼180mg% and 8∼12°Brix) than in kale(101mg% and 7oBrix). In mineral analysis by AA and ICP Mass, 'Pilgrim' showed a higher content of K, Fe, Mn, Zn and Se. In separate experiment, sensory scores of bitter taste and astringent taste were lower and scores of over all taste and over all acceptability, higher in broccoli('1243') juice than those in kale juice.

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