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Determination Method of Puerarin and Daidzin from Puerariae Radix by Reversed-Phase HPLC with Pulsed Amperometric Detection (RP-HPLC-PAD를 이용한 갈근(葛根)과 갈근(葛根) 함유 처방의 Puerarin과 Daidzin 분석)

  • Chiao, Chen-Ying;Kwon, Ha-Jeong;Jeong, Ji-Seon;Lee, Je-Hyun;Hong, Seon-Pyo
    • The Korea Journal of Herbology
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    • v.23 no.4
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    • pp.171-177
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    • 2008
  • Objectives: This study presents a reversed-phase high-performance liquid chromatography- pulsed amperometric detection(RP-HPLC-PAD) method for the determination of puerarin and daidzin in Puerariae Radix extract and Chinese medicinal preparations. Methods: Chromatographic separation was performed using a 10% acetonitrile with a reversed-phase column(Unison UK-C18, $100mm{\times}2.0mm$ I.D.; $3{\mu}m$). The analyses were detected by pulsed amperometric detector(PAD) in alkaline conditions by combining with post-column NaOH solution. Geniposide was used as an internal standard. Results: The limit of detection(S/N=3) and the limit of quantification(S/N=10) were 0.025 ng, 0.075 ng for puerarin, and 0.05 ng, 0.15 ng for daidzin, respectively. The intra- and inter-day precisions(RSDs) were less than 6.5% and average recoveries of puerarin were 99.7-101.3% and those of daidzin were 101.0-102.8%. Conclusions: According to above results, we developed a determination method for puerarin and daidzin in Puerariae Radix with high sensitivity and selectivitely.

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Characteristics of Immobilized Rhodopseudomonas sp. for Wastewater Treatment (폐수처리를 위한 고정화 Rhodopseudomonas sp.균의 특성)

  • 이범규;김상희;김중균
    • Journal of Life Science
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    • v.9 no.3
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    • pp.268-275
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    • 1999
  • Rhodopseudomonas sp. was immobilized in three supports(agar, k-carrageenan, and PVA) in order to remove nitrate in wastewater coming from fish farm. Among them 3% agar was the most suitable support when denitrification rate and bead durability were tested. Optimum bead size was 4mm-diameter when the substrate transfer into the bead and shear stress for bead were considered, and optimum cell loading was 25mg dry $cells/cm^2$gel gel. Ethanol was the best as a carbon source, and optimum C:N ratio, temperature and pH were 1.5:1, $31^{\circ}C$,, and 6, respectively. Under these conditions the maximum denitrification rate in synthetic wastewater was $$345{\MU}{\ell};N_2/Cm^3 gel{\cdot}hr;and that in modified MYC medium was 450{\MU}{\ell}};N_2/Cm^3 gel{\cdot}hr $$.

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Effect of Methionine Sulfoximine in nitrogenase activity by ammonia and glutamine in Rhodopseudomonas sphaeroides (암모니아와 glutamine에 의한 Rhodopseudomonas sphaeroides의 질소 고정 효소활성에 미치는 Methionine Sulfoximine의 영향)

  • 이혜주
    • Korean Journal of Microbiology
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    • v.26 no.3
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    • pp.215-222
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    • 1988
  • The effect of ammonia and glutamine on nitrogenase activity of Rhodopseudomonas sphaeroides was examined. The nitrogenase activity of this strain was inhibited by ammonia and glutamine. When ammonia and glutamine were exhausted, nitrogenase activity promptly resumed at its original rate. Methionine sulfoximine (MSX), irreversible glutamine synthetase (GS) inhibitor, is a structural analogue of glutamate. MSX was used in order to know whether the nitrogenase activity was inhibited by ammonia and glutamine directly or not. The ability of MSX to prevent nitrogenase switch-off by ammonia was found to be dependent upon the phase of culture. When the cells were sampled after 12 hour culture, $500{\mu}M$ MSX would not prevent the nitrogenase switch-off by ammonia. Twenty one percents of GS actibity was inhibited by $500{\mu}M$ of MSX and concentration of released ammonia decreased. But nitrogenase activiy was still inhibited by ammonia. However, nitrogenase switch-off after 20 hours would be prevented by $100{\mu}M$ of MSX. On the other hand, GS activity was ingibited completely by $100{\mu}M$ MSX and concentration of released ammonia somewhat increased. But nitrogenase activity was not inhibited. The data indicated that the inhibition of in vivo nitrogenase actibity of Rp. sphaeroides by ammonia seemed to be mediated by products of ammonia assimilation rather than by ammonia itself.

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Effect of Drug Eluting Uniformity for Biodegradable Stent by Solid Freeform Fabrication (쾌속조형기법을 이용한 생분해성 스텐트용 메쉬필름의 약물방출거동 효과)

  • Cheong, Sin Young;Kim, Yang Eun;Koh, Young Joo;Shin, Wang Soo;Lee, Jun Hee;Kim, Wan Doo;Yoo, Young Eun;Park, Su A
    • Polymer(Korea)
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    • v.38 no.1
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    • pp.93-97
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    • 2014
  • Biodegradable drug-eluting stent has dual functions of supporting the lumen and treating internal tumor preventing the restenosis by releasing drug. In this study, the polycaprolactone (PCL) based three dimensional (3D) mesh loaded with paclitaxel (PTX) was presented by rapid prototyping (RP) technique of solid freeform fabrication (SFF) for biodegradable drug-eluting stent application. PCL has many advantageous properties such as good biocompatibility, good mechanical properties, and good drug permeability. PTX is widely used in the cancer treatment by inhibiting tumor cell proliferation. Analytical methods of HPLC and NMR were used for simultaneous quantification of PTX. Scanning electron microscopy (SEM) was performed to observe the architecture and morphologies of 3D mesh. The cytotoxicity assay results indicated released PTX's biological activity. This study provided that PCL based 3D mesh loaded with PTX by RP technique has great potential for biodegradable drug-eluting stent application.

Dynamic Model Considering the Biases in SP Panel data (SP 패널데이터의 Bias를 고려한 동적모델)

  • 남궁문;성수련;최기주;이백진
    • Journal of Korean Society of Transportation
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    • v.18 no.6
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    • pp.63-75
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    • 2000
  • Stated Preference (SP) data has been regarded as more useful than Revealed Preference (RP) data, because researchers can investigate the respondents\` Preference and attitude for a traffic condition or a new traffic system by using the SP data. However, the SP data has two bias: the first one is the bias inherent in SP data and the latter one is the attrition bias in SP panel data. If the biases do not corrected, the choice model using SP data may predict a erroneous future demand. In this Paper, six route choice models are constructed to deal with the SP biases, and. these six models are classified into cross-sectional models (model I∼IH) and dynamic models (model IV∼VI) From the six models. some remarkable results are obtained. The cross-sectional model that incorporate RP choice results of responders with SP cross-sectional model can correct the biases inherent in SP data, and also the dynamic models can consider the temporal variations of the effectiveness of state dependence in SP responses by assuming a simple exponential function of the state dependence. WESML method that use the estimated attrition probability is also adopted to correct the attrition bias in SP Panel data. The results can be contributed to the dynamic modeling of SP Panel data and also useful to predict more exact demand.

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Acibenzolar-S-Methyl(ASM)-Induced Resistance against Tobamoviruses Involves Induction of RNA-Dependent RNA Polymerase(RdRp) and Alternative Oxidase(AOX) Genes

  • Madhusudhan, Kallahally Nagendra;Deepak, Saligrama Adavigowda;Prakash, Harishchandra Sripathi;Agrawal, Ganesh Kumar;Jwa, Nam-Soo;Rakwal, Randeep
    • Journal of Crop Science and Biotechnology
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    • v.11 no.2
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    • pp.127-134
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    • 2008
  • Tobamoviruses are the major viral pathogens of tomato and bell pepper. The preliminary results showed that Acibenzolar-Smethyl(ASM; S-methylbenzo(1,2,3) thiadiazole-7-carbothiate) pre-treatment to tomato and tobacco plants reduces the concentration of Tomato mosaic tobamovirus(ToMV) and Tobacco mosaic tobamovirus(TMV) in tomato and bell pepper seedlings, respectively. Pre-treatment of the indicator plant(Nicotiana glutinosa) with the ASM followed by challenge inoculation with tobamoviruses produced a reduced number and size of local lesions(67 and 79% protection over control to TMV and ToMV inoculation, respectively). In order to understand the mechanism of resistance the gene expression profiles of antiviral genes was examined. RT-PCR products showed higher expression of two viral resistance genes viz., alternative oxidase(AOX) and RNA dependent RNA polymerase(RdRp) in the upper leaves of the ASM-treated tomato plants challenge inoculation with ToMV. Further, the viral concentration was also quantified in the upper leaves by reverse transcription PCR using specific primer for movement protein of ToMV, as well as ELISA by using antisera against tobamoviruses. The results provided additional evidence that ASM pre-treatment reduced the viral movement to upper leaves. The results suggest that expressions of viral resistance genes in the host are the key component in the resistance against ToMV in the inducer-treated tomato plants.

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Natural Dyeing of Silks Dyed Singly with Caesalpinia sappan, Dactylopius coccus, Combination-Dyed with Caesalpinia sappan and Phellodendron Amurense Rupr., and with Dactylopius coccus and Phellodendron Amurense Rupr. (소목 및 코치닐로 단독 염색한 견직물, 그리고 소목과 황벽 및 코치닐과 황벽으로 복합 염색한 견직물의 천연염색)

  • Jung, Jin Soun
    • The Journal of the Convergence on Culture Technology
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    • v.6 no.4
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    • pp.275-281
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    • 2020
  • We studied the dyeability and functionality of silks dyed singly with Caesalpinia sappan, Dactylopius coccus, combination-dyed with Caesalpinia sappan and Phellodendron Amurense Rupr., and with Dactylopius coccus and Phellodendron Amurense Rupr.. The color of silk dyed singly with Caesalpinia sappan was Red with Hue of 2.2 R, the color of silk dyed singly with Dactylopius coccus was Reddish Purple with Hue of 3.0 RP, The color of silk combination-dyed with Caesalpinia sappan and Phellodendron Amurense Rupr. was 2.7YR being in the Orange family, and the color of silk combination-dyed with Dactylopius coccus and Phellodendron Amurense Rupr. was Red with Hue of 4.3R. In the case of antimicrobial activity, except for silk dyed only with Dactylopius coccus, the other three types of dyed silk showed excellent antimicrobial activity against Staphylococcus aureus achieving higher than 91.1% of bacteria reduction rate. Against Klebsiella pneumoniae, silk combination-dyed with Dactylopius coccus and Phellodendron Amurense Rupr., and with Caesalpinia sappan and Phellodendron Amurense Rupr. presented very excellent antimicrobial property achieving bacteria reduction rate of 99.9%.

Restriction map of a cryptic plasmid from Pseudomonas putida (Pseudomonas putida로 부터 분리한 cryptic플라스미드의 제한효소지도)

  • 김훈규;고상균;이영록
    • Korean Journal of Microbiology
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    • v.24 no.1
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    • pp.7-11
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    • 1986
  • We screened lysates of the laboratory strains of pseudomonads utilizing hydrocarbon by agarose gel electrophoresis and cesium chloride-ethidium bromide equilibrium centrifugation, to find an intrinsic plasmid as a vector and to examine the relationship between the plasmid and hydrocarbon degradation. Only one strain from the examined strains, Pseudomonas putida KU190, contained a plasmid. We named the plasmid pKU41. The molecular size of pKU41 was determined as 41kb, using covalently closed circular forms of RP4 and pSY343 as standard size markers. The restriction sites of pKU41 for BamHI, BglII, EcoRI, HindIII, and SalI were 3, 1, 3, 6 and more than 13, respectively. With double or triple digestion, restriction map of pKU41 was constructed for BamHI, BglII and HindIII. For elucidation on the biological function of the plasmid, test was conducted on the ability of hydrocarbon utilization of the host strain but no apparent relationship was observed.

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Identification and Characterization of a Novel Antioxidant Peptide from Bovine Skim Milk Fermented by Lactococcus lactis SL6

  • Kim, Sang Hoon;Lee, Ji Yoon;Balolong, Marilen P.;Kim, Jin-Eung;Paik, Hyun-Dong;Kang, Dae-Kyung
    • Food Science of Animal Resources
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    • v.37 no.3
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    • pp.402-409
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    • 2017
  • A novel peptide having free radical scavenging activity was separated, using an on-line high-performance liquid chromatography (HPLC) - ABTS screening method, from bovine skim milk fermented by Lactococcus lactis SL6 (KCTC 11865BP). It was further purified using reverse phase-HPLC (RP-HPLC) and sequenced by RP-HPLC-tandem mass spectrometry. The amino acid sequence of the identified peptide was determined to be Phe-Ser-Asp-Ile-Pro-Asn-Pro-Ile-Gly-Ser-Glu-Asn-Ser-Glu-Lys-Thr-Thr-Met-Pro-Leu-Trp (2,362 Da), which is corresponding to the C-terminal fragment of bovine ${\alpha}_{s1}$-casein (f179-199). The hydroxyl radicals scavenging activity ($IC_{50}$ $28.25{\pm}0.96{\mu}M$) of the peptide chemically synthesized based on the MS/MS data showed a slightly lower than that of the natural antioxidant Trolox ($IC_{50}$ $15.37{\pm}0.52{\mu}M$). Furthermore, derivatives of the antioxidant peptide were synthesized. The antioxidative activity of the derivatives whose all three proline residues replaced by alanine significantly decreased, whereas replacement of two proline residues in N-terminal region did not affect its antioxidative activity, indicating that $3^{rd}$ proline in C-terminal region is critical for the antioxidative activity of the peptide identified in this study. In addition, N-terminal region of the antioxidant peptide did not show its activity, whereas C-terminal region maintained antioxidative activity, suggesting that C-terminal region of the peptide is important for antioxidative activity.

Method development and validation of spectrophotometric and RP-HPLC methods for simultaneous estimation of spironolactone and furosemide in bulk and combined tablet dosage forms

  • Chavan, Rohankumar R.;Bhinge, Somnath D.;Bhutkar, Mangesh A.;Randive, Dheeraj S.;Salunkhe, Vijay R.
    • Analytical Science and Technology
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    • v.34 no.5
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    • pp.212-224
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    • 2021
  • The intent of the present work was to develop a simple, sensitive, accurate, precise, rapid and economical UV- spectrophotometric and reverse phase high pressure liquid chromatographic method for the simultaneous estimation of Spironolactone and Furosemide in bulk and combined tablet dosage forms. UV-Spectrophotometry was carried out by simultaneous equation method using 0.02 M potassium dihydrogen phosphate buffer pH 3.5: Acetonitrile (50:50) v/v as a solvent. The linearity range was 2-14 ㎍ mL-1 for Spironolactone and Furosemide with a correlation coefficient > 0.99. The chromatographic separation was achieved on 250 mm × 4.6 mm, hypersil BDS C18 column with particle size 5 ㎛, by using an isocratic mixture of 0.02 M potassium dihydrogen phosphate buffer pH 3.5: Acetonitrile: tert butyl methyl ether (49:50:1) v/v/v as a solvent at a flow rate of 1 mL min-1 and UV detection was carried out at 254 nm. The retention time were observed to be 3.666 and 6.661 minutes for Furosemide and Spironolactone respectively. The two developed methods were validated according to the ICH guidelines for accuracy, precision, linearity, LOD, LOQ and were found to be within the limits. It can be concluded that these two methods could be successfully used for the simultaneous estimation of Spironolactone and Furosemide in bulk and combined tablet dosage forms.