• Title/Summary/Keyword: LC20

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Stable Maintenance of Recombinant Plasmid Containing trp $^+$ Operon in E. coli Cultures by the phe W$^+$ -pheS$^{t8}$ System (대장균 배양 중 phe W$^+$-pheS-$^{-ts}$ System에 의한 재조합 trp$^+$ 플라스미드의 안정적 유지)

  • 강충민;최장원;이세영
    • Microbiology and Biotechnology Letters
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    • v.18 no.1
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    • pp.89-93
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    • 1990
  • To improve the stability of recombinant pBR322-trip$^+$ plasmid (pLTW24) in E. coli culture, a positive selection system was devised. A DNA fragment containing pheW$^+$ gene (a structural gene for tRNA$^{phe}$ was isolated and inserted into the pBR322-trip$^+$ plasmid(pLTP24). A temperature sensitive host strain. LC901-pheS$^{-ts}$, was constructed for this plasmid by transducing pheS$^{-ts}$ allele (phenylalanyl-tRNA synthetase) to E. coli LC901 using P1kc bacteriophage. The LC901-pheS$^{-ts}$ cells were unable to grow at a restrictive temperature when they had lost the pBR322 :: pheW$^+$ (pLTP24) plasmid. The effects of pheW$^+$ gene on the plasmid stability and the expression level of trip$^+$ gene in LC901-pheS$^{-ts}$ strain were investigated. The proportion of Trip$^+$ colonies among LC901-pheS$^{-ts}$ strain carrying plasmid pLTP24 was 99%, whereas that of LC901 strain carrying plasmid pLTW24 was 7% at the end of 20 generations. After 100 generations of growth, the strain LC901-pheS$^{-ts}$ carrying plasmid pLTP24 showed little loss of plasmids. While the majority of plasmid pLTW24 in LC901 strain were lost in the same period. The activities of tryptophan synthetase (T. Sase) and anthranilate synthetase (A. Sase) in LC901 strain carrying pLTW24 were about 1.2 times and 1.8 times respectively of those in LC901-pheS$^{-ts}$ strain carrying pLTP24.

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Variation of Antimicrobial Susceptibility and Phage Types of Staphyloccus aureus Derived from Different Environmental Sources (서식환경에 따른 황색포도구균의 항균제감수성 및 Phage형별의 차이)

  • Cho, Dong-Taek;Lee, Yoo-Chul;Kim, Jin-Mo
    • The Journal of the Korean Society for Microbiology
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    • v.20 no.1
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    • pp.1-11
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    • 1985
  • A total of 211 strains of Staphylococcus aureus which included 118 strains isolated from various clinical specimens of admitted patients of University Hospital with systemic or severe cases of infection and 93 strains from infected skin diseases of out-patients of dermatology clinic located in rural area, were tested for the antimicrobial susceptibility to the 12 drugs of common use and the phage typing. An these were subjected to the study of plasmid profile analysis for the molecular epidemiology of nosocomial infections. University Hospital(UH) isolates showed higher frequency of resistance than local clinic(LC) isolates against 10 drugs excluding tetracycline(Tc), and trimethoprim(Tp). The MIC of UH isolates were above than $128{\mu}g/ml$ against 9 drugs except Tc, gentamicin(Gm), and Tp, but LC isolates did not show such a high level of MIC. There was difference of MIC needed to inhibit 90% of strains(MIC90) against each drugs tested between two groups of UH and LC isolates. UH isolates showed 2 to 4 times higher value of MIC90 by two-fold serial dilution of drug concentration than LC isolates. Tp was considered as an effective drug in treatment of staphylococcal infections whereas ampicillin and Gm were appeared to be ineffective. Seventy-three strains(61.9%) of UH isolates and 70(69.9%) of LC were typable with phages from Colindale Reference Laboratory. The prevailing phage type of UH isolates belonged to lytic group II were 27 strains(22.9%) and those of LC isolates belonged to lytic group II were 23 strains(24.7%). Thirteen strains(11.0%) of UH isolates were multiply resistant to more than 5 drugs to 10 drugs but none of LC isolates. Through the lysis method of Kado and Liu followed by agarose gel electrophoresis, none of 211 strains showed plasmid profile. These results were confirmed by re-examination through the method of Birnboim and Doly.

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Evaluation of the Method for Total Homocysteine in Plasma Using LC-MS/MS (LC-MS/MS를 이용한 Homocysteine 측정과 그 유용성 평가)

  • Jun, Sun-Hee;Lim, Mi-Suk;Jung, Yong-Sun;Song, Jung-Han
    • Korean Journal of Clinical Laboratory Science
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    • v.37 no.1
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    • pp.22-26
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    • 2005
  • Total homocysteine is now considered a risk factor for cardiovascular diseases. I increased interest has led to a multitude of studies requiring the determination of total homocysteine in conjunction with other factor. There are various methods for measuring total homocysteine, including HPLC, FPIA, GC-MS and LC-MS/MS. The most recent method for measuring total homocysteine uses a deuterium-labelled internal standard and tandem mass spectrometry. This development requires no derivatization and therefore leads to an increase in sample throughput compared to other techniques. We have evaluated the method for homocysteine by the LC-MS/MS method, and the correlation between the FPIA method and the LC-MS/MS method. The standard curve (0, 5, 10, 20, 50, 100 uM) was linear over the range examined (up to 100 uM). The lower limit of quantification (CV < 10 %) was 0.5 uM/L and the lower limit of detection (S/N >3) was 0.1 uM/L. Intra-assay variation and inter-assay variation were both <6 %. The comparision study for homocysteine concentration showed good correlation (r=0.9684) between the FPIA method and LC-MS/MS methods. Our conclusion is that the method showed relatively good precision, and was rapid and accurate.

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Comparative Study of LC Scheme with Some Conventional Schemes by Truncation Error Analysis (선형특성 (LC) 법과 그 외 고전적 방법들과의 절단오차 분석에 의한 비교연구)

  • Kim, Chan-Hyeong;Kim, Jong-Kyung;Yook, Chong-Chul
    • Nuclear Engineering and Technology
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    • v.20 no.3
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    • pp.179-188
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    • 1988
  • A recently developed spatial differencing scheme, Linear Characteristic (LC) scheme is compared with some traditionally used schemes such as Step Difference (SD), Diamond Difference (DD), and Step Characteristic (SC) scheme by analyzing the truncation error calculated numerically in slab geometry. Those four candidate schemes are applied to one simple source sink problem and two criticality problems (one is calculation of multiplication factor and the other is slab critical half thickness). The calculated results are then examined by some equitable measures of error. It is concluded that the LC scheme is terribly more powerful than any other candidate scheme that has been prevalent up to the present time. Moreover, the LC scheme estimates integral parameter such as multiplication factor and critical half thickness much more efficiently than SD or SC scheme. This is due to the fact that the fortuitous error cancellation, which occurs when the deviations of cell average flux are summed over the whole gamut of spatial meshes, happens much more favorably to the LC scheme.

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Fodder Productivity and Growth Persistency of Three Local Cassava Varieties

  • Tung, C.M.;Liang, J.B.;Tan, S.L.;Ong, H.K.;Jelan, Z.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.9
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    • pp.1253-1259
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    • 2001
  • Three cassava varieties, namely MM 92 (MM), Black Twig (BT) and Local (LC), were arranged in a randomized complete block design to evaluate their dry fodder and crude protein (CP) productivity as well as growth persistency. Cassava plants grown in small plots of $5m{\times}10m$ at a planting distance of $25cm{\times}25cm$ were harvested every 6 weeks starting from 3 months after planting. Dry fodder yields of MM, BT and LC over the 8 harvests were 8.55, 8.01 and 6.15 t/ha, respectively. All varieties produced more leaves than stems with average leaf:stem ratios of 5, 5.9 and 4.8 for MM, BT and LC, respectively. In terms of CP production, MM was the highest yielder (272 kg/ha/harvest), followed by BT and LC (238 and 184 kg/ha/harvest, respectively). The total accumulative CP amounts over the 8 harvests were 2179, 1903 and 1474 kg/ha for MM, BT and LC, respectively. The mortality rates were 9.91, 14.01 and 13.98% for MM, BT and LC, respectively. Phosphorus content was more stable than potassium content during defoliation. MM, BT and LC had whole plant phosphorus contents of 0.41, 0.41 and 0.39%, respectively; whole plant potassium contents were 1.25, 1.38 and 1.20%.

Isolation and Structural Identification of Antioxidant Substances from Ethyl Acetate Extract of Conyza canadensis (망초(Conyza canadensis) Ethyl Acetate 추출물의 항산화성 물질의 분리와 동정)

  • Hyun Sook Song
    • Journal of Naturopathy
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    • v.12 no.1
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    • pp.7-15
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    • 2023
  • Background: As a result of analyzing the components of wild Conyza canadensis, it contains physiologically active ingredients, so it is necessary to identify the compound. Purposes: It was to study the compound's molecular structure; a previous study showed that C. canadensis contains antioxidant substances. Methods: The ultrasonic pulverized lysate of C. canadensis stem and leaves was first extracted with 90% methanol and then five organic solvents. Next, the extracts was fractionated by HPLC, LC/MS chromatography, and NMR analyzers identified the molecular structure. Results: 100 g of dry C. canadensis was sonicated in 90% methanol and concentrated under reduced pressure to 11.96 g of a crude extract. Then, this crude was extracted with five types of solvents to obtain 123.8 mg of n-hexane, 448.2 mg of dichloromethane, 1047.7 mg of ethyl acetate (EA), 2563.8 mg of butanol, and 7.04 g of water. The EA extracts were fractionated by LC-MS and then re-fractionated to obtain F1 to F20. Next, the F15 was further fractionated to obtain nine fine fractions. Finally, the F17 fraction was re-fractionated to obtain ten fine fractions. As a result of LC-MS and NMR spectrometer analysis of the F15-7, the structure of this compound was confirmed as 3,5-dicaffeoylquinic acid. As a result of examining the structures of the F17-4 and F17-5 fractions, Quercetin-3-o-β-galactose was identified. In addition, the form of the F17-10 was confirmed to be 1,3,4-tri-caffeoylquinic acid. Conclusions: This study demonstrated that C. canadensis contained phenolic antioxidants, and its utilization may be expected.

Efficacy of Some Insecticides against Ectinohoplia rufipes (Coleoptera: Scarabaeidae) on Different Host Plant Species (기주 식물에 따른 몇 가지 살충제의 주황긴다리풍뎅이, Ectinohoplia rufipes(Coleoptera: Scarabaeidae)에 대한 살충효과)

  • 최우근;이동운;이승욱;추호렬;박정규;사공영보
    • Asian Journal of Turfgrass Science
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    • v.15 no.3
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    • pp.119-126
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    • 2001
  • The susceptibility of Ectinohoplia rufipes, insect pest of landscaping trees in golf courses to registered insecticides in golf courses was investigated. In addition, effect of host plants, Ainus firma, Prunus serrulata var. spontanae, and Osmantus asiaticus on susceptibility of E. rufipes were tested in laboratory with efficacy of some insecticides in laboratory and fields. The insecticides used in experiments were cabaryl 40%+phosalone 20% WP, chlorpyrifos-methyl 25% EC, chlorpyrifos 20% + diflubenzuron 7% n, deltamethrin 1% EC, ethopenprox 8% +diazinon 25% WP, and fenitrothion 50% EC. Mortalities of E.rufipes were 100% except chlorpyrifos-methyl. when A. firma leaves dipped in recommended concentration of given insecticides were supplied as food to adult of E. rufipes. LC$_{50}$s of insecticides were different depending on host plants, that is, those of deltamethrin were 0.21 ppm in Ο. asiaticus, 0.45 ppm in P. serrulata var. spontanae, and 0.77 ppm in A. firma. LC$_{50}$s of fenitrothion were 24.1 ppm, 64.0 ppm, and 11.0 ppm and those of chlorpyrifos-methyl were 97.2 ppm, 40.6 ppm, and 29.6 ppm, respectively. LC$_{50}$ was the lowest in Ο. asiaticus and that of chlorpyrifos-methyl and fenitrothion was the lowest in A. firma. Deltamethrin was the most effective against adult of E. rufipes in field like in laboratory and persisted in two weeks.weeks.

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Sensitive Determination of Felodipine in Human Plasma by LC-MS/MS

  • Kim, Ho-Hyun;Roh, Hyeon-Jin;Lee, He-Joo;Han, Sang-Beom
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.283.2-283.2
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    • 2003
  • This study established a highly sensitive novel quantification method for detecting felodipine in human plasma using LC-MS/MS. The mobile phase used after degassing was composed of 1 mM ammonium acetate and acetonitrile (20:80, pH 6.0), with flow rate of 200 uL/min. One mL plasma were pipetted into glass tubes and spiked with 0.1 mL of internal standard solution. (omitted)

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Determination of Glimepiride in Human Plasma by LC-MS/MS

  • Kim, Ho-Hyun;Roh, Hyeong-Jin;Lee, Hee-Joo;Han, Sang-Beom
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.282.1-282.1
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    • 2003
  • This study established a sensitive novel Quantification method for detecting glimepiride in human plasma using LC-MS/MS for pharmacokinetic studies. The mobile phase used after degassing was composed of 10 mM ammonium acetate and acetonitrile (20:80, pH 3.0), with flow rate of 200uL/min. One mL plasma were pipetted into glass tubes and spiked with 0.1 mL of internal standard solution. (omitted)

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Multiresidue Analysis of 124 Pesticides in Soils with QuEChERS extraction and LC-MS/MS (QuEChERS 및 LC-MS/MS를 이용한 토양 중 124종 잔류농약다성분 분석법)

  • Gwon, Ji-Hyeong;Kim, Taek-Kyum;Seo, Eun-Kyung;Hong, Su-Myeong;Kwon, Hye-Yong;Kyung, Ki-Sung;Kim, Jang-Eok;Cho, Nam-Jun
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.296-313
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    • 2014
  • A QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) multiresidue method was developed for the simultaneous analysis of 124 pesticides in soil by LC-MS. The procedure involved liquid extraction of soil immersed with 0.2N $NH_4Cl$ by acetonitrile with 1% acetic acid, followed by anhydrous $MgSO_4$ and sodium acetate, and dispersive SPE cleanup with $MgSO_4$, primary secondary amine (PSA) and $C_{18}$. The extracts were analyzed with LC-MS/MS in ESI positive mode. Standard calibration curves were made by matrix matched standards and their correlation coefficients were higher than 0.99. Recovery studies for the validation were carried out using two type soils, loam and sandy loam, at four concentration levels (0.005, 0.01, 0.02, and 0.1 mg/kg). The recoveries of pesticides were in the range of 70-120% with < 20% RSD except 4 pesticides, Benfuracarb, Ethiofencarb, Pymetrozine, and Pyrethrin. This result indicated that the method using QuEChERS and LC-MS/MS could be applied for the simultaneous determination of pesticide residues in soils.