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The Biological Degradation of High Concentration of Trichloroethylene (TCE) by Delftia acidovornas EK2 (Delftia acidovorans EK2에 의한 고농도 Trichloroethylene (TCE)의 생물학적 분해 특성)

  • Park, Woo-Jung;Lee, Sang-Seob
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.183-191
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    • 2010
  • In this study, we isolated 179 bacterial strains using benzene, phenol, ethylbenzene, aniline, cumene, toluene as growth substrate from TCE contaminated soils and wastewaters. All the 179 strains were screened for TCE (30 mg/L) removal (growth substrate 0.2 g/L, $30^{\circ}C$, pH 7, cell biomass 1.0 g/L (w/v)) under aerobic condition for 21 days. EK2 strain using aniline showed the highest removal efficiency (74.4%) for TCE degradation. This strain was identified as Delftia acidovorans as the results of API kit, 16S rDNA sequence and fatty acid assay. In the batch culture, D. acidovorans EK2 showed the bio-degradation for TCE in the various TCE concentration (10 mg/L to 200 mg/L). However, D. acidovorans EK2 did not show the bio-degradation in the TCE 250 mg/L. D. acidovorans EK2 also show the removal efficiency (99.9%) for 12 days in the low concentration (1.0 mg/L). Optimal conditions to degrade TCE 200 mg/L were cell biomass 1.0 g/L (w/v), aniline 0.5 g/L, pH 7 and $30^{\circ}C$. Removal efficiency and removal rate by D. acidovorans EK2 strain was 71.0% and 94.7 nmol/h for 21 days under optimal conditions. Conclusion, we expect that D. acidovorans EK2 may contribute on the biological treatment in the contaminated soil or industrio us wastewater.

A Study of Reportable Range Setting through Concentrated Control Sample (약물검사에서 관리시료의 농축을 이용한 보고 가능 범위의 설정에 대한 연구)

  • Chang, Sang Wu;Kim, Nam Yong;Choi, Ho Sung;Park, Yong Won;Yun, Keun Young
    • Korean Journal of Clinical Laboratory Science
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    • v.36 no.1
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    • pp.13-18
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    • 2004
  • This study was designed to establish working range for reoportable range in own laboratory in order to cover the upper and lower limits of the range in test method. We experimented ten times during 10 days for setting of reportable range with between run for method evaluation. It is generally assumed that the analytical method produces a linear response and that the test results between those upper and lower limits are then reportable. CLIA recommends that laboratories verify the reportable range of all moderate and high complexity tests. The Clinical Laboratory Improvement Amendments(CLIA) and Laboratory Accreditation Program of the Korean Society for Laboratory Medicine states reportable range is only required for "modified" moderately complex tests. Linearity requirements have been eliminated from the CLIA regulations and from others accreditation agencies, many inspectors continue to feel that linearity studies are a part of good lab practice and should be encouraged. It is important to assess the useful reportable range of a laboratory method, i.e., the lowest and highest test results that are reliable and can be reported. Manufacturers make claims for the reportable range of their methods by stating the upper and lower limits of the range. Instrument manufacturers state an operating range and a reportable range. The commercial linearity material can be used to verify this range, if it adequately covers the stated linear interval. CLIA requirements for quality control, must demonstrate that, prior to reporting patient test results, it can obtain the performance specifications for accuracy, precision, and reportable range of patient test results, comparable to those established by the manufacturer. If applicable, the laboratory must also verify the reportable range of patient test results. The reportable range of patient test results is the range of test result values over which the laboratory can establish or verify the accuracy of the instrument, kit or test system measurement response. We need to define the usable reportable range of the method so that the experiments can be properly planned and valid data can be collected. The reportable range is usually defined as the range where the analytical response of the method is linear with respect to the concentration of the analyte being measured. In conclusion, experimental results on reportable range using concentrated control sample and zero calibrators covering from highest to lowest range were salicylate $8.8{\mu}g/dL$, phenytoin $0.67{\mu}g/dL$, phenobarbital $1.53{\mu}g/dL$, primidone $0.16{\mu}g/dL$, theophylline $0.2{\mu}g/dL$, vancomycine $1.3{\mu}g/dL$, valproic acid $3.2{\mu}g/dL$, digitoxin 0.17ng/dL, carbamazepine $0.36{\mu}g/dL$ and acetaminophen $0.7{\mu}g/dL$ at minimum level and salicylate $969.9{\mu}g/dL$, phenytoin $38.1{\mu}g/dL$, phenobarbital $60.4{\mu}g/dL$, primidone $24.57{\mu}g/dL$, theophylline $39.2{\mu}g/dL$, vancomycine $83.65{\mu}g/dL$, valproic acid $147.96{\mu}g/dL$, digitoxin 5.04ng/dL, carbamazepine $19.76{\mu}g/dL$, acetaminophen $300.92{\mu}g/dL$ at maximum level.

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Protective Effect of Mixed Extract including Hovenia dulcis Thunberg against Chronic Ethanol Treatment-induced Cytotoxicity in a Brain and Liver Tissue (만성 알코올 섭취로부터 유도되는 뇌 및 간 조직 독성에 대한 지구자 혼합 추출물의 보호 효과)

  • Kim, Jong-Min;Park, Seon-Kyeong;Guoa, Tian-Jiao;Kang, Jin-Yong;Ha, Jeong-Su;Lee, Du-Sang;Kwon, O-Jun;Lee, Uk;Heo, Ho Jin
    • Journal of agriculture & life science
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    • v.50 no.2
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    • pp.125-138
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    • 2016
  • To assess the industrial possibility of mixed-extracts containing Hovenia dulcis Thunberg and 12 different botanical ingredients, a protective effect was confirmed in the chronic ethanol-induced the liver, brain, and blood injury in mouse. Blood glucose levels of the normal control group(NG) and ethanol administration group(EG) were respectively 119.43mg/dL and 305.25mg/dL, and the mixed-extracts administration group(100, 200mg/kg body weight + 25% ethanol 5g/kg body weight respectively; ME100 & ME200) were decreased to 272.76mg/dL and 234.60mg/dL. Blood ethanol contents were decreased in ME100 and ME200(3.85mg/dL, 3.08mg/dL) compared to EG(4.08mg/dL), and blood acetaldehyde contents were also decreased in ME(15.76mg/dL, 15.16mg/dL) compared to EG(18.72mg/dL). The contents of hepatotoxic indicators such as glutamine pyruvic transaminase(GPT) and glutamic oxaloacetic transaminase (GOT), nephrotoxic indicators such as blood urea nitrogen(BUN), and creatine(CRE), and total cholestero(TCHO), and triglyceride(TG) in mouse blood serum were significantly decreased in the ME compared to EG. The acetylcholinesterase(AChE) activity of ME(109.00% and 108.47%, respectively) in mouse brain tissues was decreased in ME compared to EG(116.10%). Finally, ME was remarkable in vivo antioxidant activities in the mouse liver and brain tissues by superoxide dismutase(SOD), oxidized glutathione(GSH)/total GSH ratio and the malondialdehyde (MDA) assay. Therefore, the mixed-extracts was considered to be effective a high value food with protective effect against chronic ethanol traetment-induced cytotoxicity in liver and brain tissues.

A Study on the Inhibition of 2-deoxy-D-Glucose Transport of the Endogenous Glucose Transporters in Spodoptera frugiperda Clone 21-AE Cells by Using Hexoses

  • Lee Chong-Kee
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.487-492
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    • 2005
  • The baculovirus/insect cell expression system is of great value in the study of structure-function relationships in mammalian glucose-transport proteins by site-directed mutagenesis and for the large-scale production of these proteins for mechanistic and biochemical studies. Spodoptera frugiperda Clone 21 (Sf2l) cells grow well on TC-100 medium that contains $0.1\%$ D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, very little is known about the properties of the endogenous sugar transporter(s) in Sf2l cells, although a saturable transport system for hexose uptake has been previously revealed in the Sf cells. In order to further examine the substrate and inhibitor recognition properties of the Sf2l cell transporter, the ability of hexoses to inhibit 2-deoxy-D-glucose (2dGlc) transport was investigated by measuring inhibition constants $(K_i)$. The $K_i's$ for reversible inhibitors were determined from plots of uptake versus inhibitor concentration. Transport was effectively inhibited by D-mannose and D-glucose. Of the hexoses tested, L-glucose had the least effect on 2dGlc transport in the Sf2l cells, indicating that the transport is stereoselective. Unlike the human HepG2 type glucose transport system, D-mannose had a somewhat greater affinity for the Sf2l cell transporter than D-glucose, implying that the hydroxyl group at the C-2 position is not necessary for strong binding. However, epimerization at the C-4 position of D-glucose (D-galactose) resulted in a dramatic decrease in affinity of the hexose for the Sf2l cell transporter. Such a lowering of affinity might be the result of the involvement of the C-4 hydroxyl in hydrogen bonding. It is therefore suggested that Sf2l cells were found to contain an endogenous sugar transport activity that in several aspects resembles the human HepG2 type glucose transporter, although the insect and human transporters do differ in their affinity for cytochalasin B.

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Effect of Medium Component on Plant Regeneration via Adventitious Bud Formation from Leaf Explant Cultures of Strawberry (Fragaria ananassa Duch.) (딸기 (Fragaria ananassa Duch.) 잎 절편체 배양으로부터 부정아 형성을 통한 식물체 재생)

  • 조덕이;소웅영;정원일
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.171-178
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    • 2001
  • This study was investigated to establish a regeneration system of plant via adventitious bud formation from leaf explant cultures of strawberry (Fragaria ananassa Duch). Effects of plant growth regulators (2,4-D, BAP), agar sucrose and myo-inositol on adventitious bud formation were investigated. When the leaf explants were cultured on MS medium supplemented with 0.1 mg/L 2,4-D and 3 mg/L BAP, the adventitious bud formation was most promoted. The adventitious bud formation was not induced from leaf explants cultured on MS medium containing 2,4-D alone. Adventitious bud formation was enhanced to almost 3 times on medium with low level of agar concentration (0.4%) in comparison with those on the medium with high level of agar (1%), but almost of shoot was vitrificated on the medium. Therefore, the normal adventitious bud formation from leaf explants was most effective on the medium containing 0.05 mg/L 2,4-D,1 mg/L BAP, 0.8% agar, 30 g/L sucrose and 100 mg/L myo-inositol. Thus, the mass propagation of healthy strawberry could be established using leaf explants.

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Effect of Plant Growth Regulators on Bulblet Formation and Plant Regeneration in Fritillaria thunbergii Miq. (패모조직배양에서 생장조절 물질이 자구형성 및 식물체 재생에 미치는 영향)

  • Park, Chul-Hyoung;Ryu, Jeom-Ho;Han, Kwang-Soo;Doo, Hong-Soo;Choi, Sun-Young
    • Korean Journal of Medicinal Crop Science
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    • v.4 no.2
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    • pp.119-125
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    • 1996
  • To improve the efficiency of mass propagation in vitro of Fritillaria thunbergii, bulb scale and nodes were cultured in LS medium supplemented with the combination of 2, 4-D and kinetin or NAA and BA. The number and size of bulb, the number of adventitious shoot, the ratio of callus formation, rooting, and the effects of light and dark on the culture, plant regeneration from calli, and the gelling substances were investigated. The combination of 2, 4-D and kinetin in media was more effective than the media of NAA and BA for the bulblet formation. The media supplemented with 2 mg/L 2, 4-D and 1 mg/L kinetin, $1{\sim}2\;mg/L$ 2, 4-D without kinetin and $1{\sim}3\;mg/L$ BAA only were effective in the adventitious shoot development. Callus formation and root formation, respectively were effective in the medium supplemented with 2mg/L 2, 4-D and 1mg/L kinetin. In bulb formation, the medium with 5 mg/L kinetin was effective, and the most of bulbs were formed from the axillary bud of node part. In bulb formation, shoot growth, callus and root formation, the light culture for 16 hours per day was better than that in the dark culture. Bulb was nicely formed in the medium with 0. 2 mg/L 2, 4-D, 1 mg/L kinetin. The medium without hormone was most effective for plant regeneration. The phytagel was more effective than agar in the medium as the gelling agent.

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L-Phenylalanine Production by Regulatory Mutants of Excherichia coli K-12 (Escherichia coli K-12 대사조절 변이주에 의한 L-페닐알라닌 생산)

  • Lee, Sae-Bae;Park, Chung;Won, Chan-Hee;Choi, Duk-Ho;Lim, Bun-San
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.174-179
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    • 1990
  • In order to overproduce L-phenylalanine, various kind of regulatory mutants were isolated from parental Escherichia coli K-12. MWEC 83 Producing 7.4g/l of L-phenylalanine has been derived as a tyrosine and tryptophan double auxotrophic mutant. To produce L-phenylalanine without adding L-tyrosine and L-tryptophan, revertant strain MWEC 101 was isolated from MWEC 83. Further various analogues and valine resistant mutants were isolated from MWEC 101. MWEC 101-5 was the most excellent strain that produced 17.9g/l of L-phenylalanine after having been cultivated for 54 hours in 15% glucose medium. It was disclosed that activities of rate-limiting enzymes including chorismate mutase and prephenate dehydratase in MWEC 101-5 were desensitized to 2mM L-phenylalanine in the enzyme reaction mixture and that activities level of 3-deoxy-D-arabino-heptulosonic acid-7-phosphate synthase and prephenate dehydratase were increased more than 20 times over those of the parental strain.

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Probiotic Characteristics of Lactobacillus acidophilus KY1909 Isolated from Korean Breast-Fed Infant (한국인 유아 분변에서 분리한 Lactobacillus acidophilus KY1909의 프로바이오틱 특성)

  • Park, Jong-Gil;Yun, Suk-Young;Oh, Se-Jong;Shin, Jung-Gul;Baek, Young-Jin
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1244-1247
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    • 2003
  • The purpose of this study was to isolate lactic acid bacteria that produced L(+) lactic acid from infant feces. Thirteen colonies were isolated with a MRS-plate containing 0.5% $CaCO_3$ to determine their ability to produce lactic acid. Based on their lactic acid production, 10 strains of Lactobacillus were identified to assess the ratio of lactate isomer using HPLC. A strain producing L-lactic acid was identified as Lactobacillus acidophilus, using API carbohydrate fermentation patterns and physiological tests, and named KY1909. The strain exhibited good acid tolerance in an artificial gastric juice as well as high bile resistance in MRS containing 0.5% bile acids. L. acidophilus KY1909 produced D(-) and L(+) lactic acid at a ratio of 6 : 94; whereas commercial strains of Lactobacillus acidophilus produced D(-) and L(+) lactic acid at a ratio of 1 : 1. These results demonstrate the L. acidophilus KY1909 can be utilized in fermented milk products and dietary supplements as a probiotic culture.

Anomalous Structure of Somatic Embryos Developed from Leaf Explant Cultures of Angelica gigas Nakai

  • Cho, Duck-Yee;Lee, Eun-Kyong;Soh, Woong-Young
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.1-5
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    • 1998
  • This study describes the effect of the growth regulators such as 2,L-D and BA, on the structural abnormalities of somatic embryos derived from leaf explants of Angelica gigas Nakai, Also, the relationship between the cotyledon number of a somatic embryo and its germinability is explored. Embryogenic calli were selected from calli formed on explants cultured on MS solid basal medium supplemented with 0.5mg/L 2,4-D, 1mg/L 2,4-D, 1mg/L plus 0.1mg/L BA, and 1 mg/L 2,4-D plus 0.5mg/L BA. Cotyledonary abnormalities were observed in somatic embryos which were developed from embryogenic calli cultured on MS medium containing 1mg/L 2,4-D for 8 weeks and then subcultured on 2,4-D free MS medium for 3 weeks. The frequency of abnormalities was as follows: 22.8% one cotyledon, 42.5% two cotyledons, 16.8% three cotyledons, 7.8% four cotyledons, 1.8% five cotyledons, and 8.2% jar shaped cotyledon. In addition, ABA treatment indicated an improvement of the somatic embryo with normal cotyledon (65.3%). ABA was important role to the high production of normal somatic embryos. Two cotyledon embryos showed germinability 77.8%. However the germinability of somatic embryos with anomalous cotyledons was prominently low: One cotyledon, 62.5%; three cotyledons, 43.3%; four cotyledons, 60%; five cotyledons, 50% and jar shaped cotyledon, zero%. Thus, germinability was essentially, inversely proportional to cotyledon number.

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Somatic Embryogenesis in Immature Zygotic Embryo Cultures of Korean Soybean (Glycine max L.) Culitivars and Effect of 2,4-Dichlorophenoxyacetic Acid on Somatic Embryo Morphology (한국 품종 대두(Glycine max L) 미숙배로부터 체세포배발생과 배지의 2,4-Dichlorophenoxyacetic Acid 농도가 체세포배의 형태에 미치는 영향)

  • Pil S Choi;Yoong Y. SOH;Duck Y.Choi;ang R. LIU
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.1
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    • pp.7-14
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    • 1994
  • Immature zygotic embryos of five Korean soybean cultivars cultured on Murashige and Skoog's (MS) medium supplemented with various concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) produced somatic embryos without forming an intervening callus. The highest frequency (up to 90%) of somatic embryo formation was obtained when zygotic embryos were cultlued on medium containing 1 to 2 mga 2, 0-D in four cultivars. However the frequency was highly variable to the cultivars. Transversely sliced primary somatic embryo halves were also capable of forming secondary embryos at frequencies of up to 70% when cultured on medium containing 0.1 to 1 mg/L 2,4-D. Somatic embryos formed on zygotic embryos cultured on medium containing 0.1 to 0.2 mg/L 2,4-D had two cotyledons more frequently than one horn-type cotyledon and those on medium containing 0.5 to 4mg/L 2,4-LD had a horm-type cotyledon at a prominently higher freequency. However somatic embryos on medium containing 10mg/L or higher concentrations of 2,4-D were usually shunted at the globular stage even after transfer to medium containing lower concentrations of 2,4-D or other growth regulators. non somatic embryos with one or two cotyledons or a hem-type cotyledon were transferred to medium containing $GA_3$, those with two cotyledons converted to plantlets at a higher frequency (25%) than the others.

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