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Comparison of Two Laccases from Trametes versicolor for Application in the Decolorization of Dyes

  • Li, Qi;Ge, Lin;Cai, Junli;Pei, Jianjun;Xie, Jingcong;Zhao, Linguo
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.545-555
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    • 2014
  • It has been previously demonstrated that laccases exhibit great potential for use in several industrial and environmental applications. In this paper, two laccase isoenzyme genes, lccB and lccC, were cloned and expressed in Pichia pastoris GS115. The sequence analysis indicated that the lccB and lccC genes consisted of 1,563 and 1,584 bp, and their open reading frames encoded 520 and 527 amino acids, respectively. They had 72.7% degree of identity in nucleotides and 86.7% in amino acids. The expression levels of LccB and LccC were up to 32,479 and 34,231 U/l, respectively. The recombinant laccases were purified by ultrafiltration and $(NH_4)_2SO_4$ precipitation, showing a single band on SDS-PAGE, which had a molecular mass of 58 kDa. The optimal pH and temperature for LccB were 2.0 and $55^{\circ}C$ with 2,2'-azinobis-[ 3-ethylbenzthiazolinesulfonic acid (ABTS) as a substrate, whereas LccC exhibited optimal pH and temperature at 3.0 and $60^{\circ}C$. The apparent kinetic parameters of LccB were 0.43 mM for ABTS with a $V_{max}$ value of 51.28 U/mg, and the Km and $V_{max}$ values for LccC were 0.29 mM and 62.89 U/mg. The recombinant laccases were able to decolorize five types of dyes. Acid Violet 43 (100 g/ml) was completely decolorized by LccB or LccC (2 U/ml), and the decolorization of Reactive Blue KN-R (100 g/ml) was 91.6% by LccC (2 U/ml). Thus, the study characterizes useful laccase isoenzymes from T. versicolor that have the capability of being incorporated into the treatment of similar azo and anthraquinone dyes from dyeing industries.

The Effect of Magnetic Field Treatment on Enhanced Exchange Coupling of $Nd_{2}Fe_{14}B/Fe_{3}B$ Magnet (자장중 열처리에 의한 $Nd_{2}Fe_{14}B/Fe_{3}B$ 초미세립 복합자성상의 자기특성 향상)

  • 박언병;양충진
    • Journal of the Korean Magnetics Society
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    • v.6 no.2
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    • pp.80-85
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    • 1996
  • An externally applied magnetic field during annealing the $Nd_{2}Fe_{14}B/Fe_{3}B$ based spring magnet was found to enhance the exchange coupling between the hard and soft magnetic grains. More than 30 % increase in remanence values for melt-spun $Nd_{4}Fe_{73.5}Co_{3}(Hf_{1-x}-Ga_{x})B_{18.5}$(x=0, 0.5, 1.0) alloys was resulted from uniform distribution of $Fe_{3}B$, $\alpha$-Fe as well as $(Nd_{2}Fe_{14}B)$, and also from reduced grain size of those phases by 20 %. The result also showed that there is an optimum grain size exhibiting a high coercivity value which will be discussed in terms of previously simulated exchange coupling parameter.

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Implementation of a FLEX Protocol Signal Processor for High Speed Paging System (고속 페이징 시스템을 위한 FLEX 프로토콜 신호처리기의 구현)

  • Gang, Min-Seop;Lee, Tae-Eung
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.38 no.1
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    • pp.69-78
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    • 2001
  • This paper presents the design and FPGA implementation of a FLEX PSP(Protocol Signal Processor) for the portable high speed paging system. In this approach, two algorithms are newly proposed for implementing the PSP which provides capabilities of the maximum 6,400bps at speed, high-channel throughput, real time error correction and an effective frame search function. One is an accurate symbol synchronization algorithm which is applied for synchronizing the interleaved 4-level bit symbols which are received at input stage of A/D converter, and the other is a modified fast decoding algorithm which is provided for realizing double error correction of (31,21)BCH signal. The PSP is composed of six functional modules, and each module is modelled in VHDL(VHSIC Hardware Description Language). Both functional simulation and logic synthesis have performed for the proposed PSP through the use of Synopsys$^{TM}$ tools on a Axil-320 Workstation, and where Altera 10K libraries are used for logic synthesis. From logic synthesis, we can see that the number of gates is about 2,631. For FPGA implementation, timing simulation is performed by using Altera MAX+ PLUS II, and its results will be also given. The PSP which is implemented in 6 FPGA devices on a PCB has been verified by means of Logic Analyzer.r.

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Effect of Silane Coupling Agent on Thermal Stability and Adhesion Properties of DGEBF Epoxy Resin (실란 커플링제에 따른 DGEBF 계열 에폭시의 열안정성 및 접착특성 평가)

  • Lee, Dong Su;Lee, Seul-Yi;Min, Byung-Gak;Seo, Young Soo;Lee, Bong Han;Park, Soo-Jin
    • Polymer(Korea)
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    • v.38 no.6
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    • pp.787-790
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    • 2014
  • In this work, the epoxy specimens were prepared from diglycidyl ether of bisphenol F (DGEBF) with silane coupling agents (3-glycidoxypropyl trimethoxysilane (GPTMS)) in different ratios. Thermal stability was studied in terms of polymer decomposition temperature (PDT), temperature of maximum rate of weight loss ($T_{max}$), integral procedural decomposition temperature (IPDT), and decomposition activation energy ($E_a$) using TGA analysis. Adhesion properties of epoxy composite specimens were measured by UTM (universal testing machine) at atmosphere temperature. In this result, the adhesion properties of DGEBF were improved by addition of silane coupling agents compared to non-treated epoxy resin. However, when the content of GPTMS agent is more than 10 phr, adhesion properties decreased with increasing GPTMS agent.

Pharmacokinetics and Bioequivalence of Haloperidol Tablet by Liquid Chromatographic Mass Spectrometry with Electrospray Ionization

  • Yun Min-Hyuk;Kwon Jun-Tack;Kwon Kwang-il
    • Archives of Pharmacal Research
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    • v.28 no.4
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    • pp.488-492
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    • 2005
  • The purpose of this study is to investigate the bioequivalence of two haloperidol 5 mg tablets, Myung In haloperidol (Myung In Pharm. Co., Ltd., test drug) and $Peridol^{R}$(Whanin Pharm. Co., Ltd., reference drug), and also to estimate the pharmacokinetic parameters of haloperidol in Korean volunteers. The bioavailability and pharmacokinetics of haloperidol tablets were examined on 24 healthy volunteers who received a single oral dose of each preparation in the fasting state in a randomized balanced 2 way crossover design. After an oral dosing, blood samples were collected for a period of 60 h. Plasma concentrations of haloperidol were determined using a liquid chromatographic electrospray mass spectrometric (LC-MS) method. The pharmacokinetic parameters were calculated with noncompartmental pharmacokinetic analysis. The geometric means of $AUC_{0-60h} and C_{max}$ between test and reference formulations were $17.21\pm8.26 ng\cdot/mL vs 17.31\pm13.24 ng\cdot/mL and 0.87\pm0.74 ng/mL vs 0.85\pm0.62 ng/mL$. respectively. The $90\%$ confidence intervals of mean difference of logarithmic transformed $AUC_{0-60h} and C_{max} were log0.9677{\sim}log1.1201 and log0.8208{\sim}log1.1981$, respectively. It shows that the bioavailability of test drug is equivalent with that of reference drug. The geometric means of other pharmacokinetic parameters ($AUC_{inf}. t_{1/2}, V_{d}/F, and CL/F$) between test drug and reference drug were $21.75\pm8.50 ng{\cdot}h/mL vs 21.77\pm15.63 ng{\cdot}h/mL, 29.87\pm8.25 h vs 29.60\pm7.56 h, 11.51\pm5.45 L vs 12.90\pm6.12 L and 0.26\pm0.09 L/h vs 0.31\pm0.17 L/h$, respectively. These observations indicate that the two formulation for haloperidol was bioequivalent and, thus, may be clinically interchangeable.

Pharmacokinetics of Verapamil and Its Major Metabolite, Norverapamil from Oral Administration of Verapamil in Rabbits with Hepatic Failure Induced by Carbon Tetrachloride

  • Choi Jun Shik;Burm Jin Pil
    • Archives of Pharmacal Research
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    • v.28 no.4
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    • pp.483-487
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    • 2005
  • The aim of this study was to investigate the pharmacokinetic changes of verapamil and its major metabolite, norverapamil, after oral administration of verapamil (10 mg/kg) in rabbits with slight, moderate and severe hepatic failure induced by carbon tetrachloride. The plasma verapamil concentrations in all groups of hepatic failure were significantly higher (p<0.01) than the control. However, the plasma norverapamil concentrations in severe hepatic failure were significantly higher (p<0.05) than the control. The peak concentrations ($C_{max}$) and the areas under the plasma concentration-time curve (AUC) of verapamil in the rabbits were significantly (p<0.01) higher than the control. The absolute bioavailability ($F_{A.B}$) and the relative bioavailability ($F_{R.B}$) of verapamil in the rabbits with hepatic failure were significantly higher ($13.6-22.2\% and 150-244\%$, respectively) than the control ($9.1\% and 100\%$, respectively). Although the AUC and $C_{max}$ of its major metabolite, norverapamil, in slight, moderate hepatic failure were not significantly lower than the control, the metabolite-parent AUC ratio in all groups of hepatic failure was decreased significantly (p<0.05, in slight group; p<0.01, in moderate and severe group) than the control. This could be due to decrease in metabolism of verapamil in the liver because of suppressed hepatic function in the hepatic failure groups because verapamil is mainly metabolized in the liver. From our data, it would seem appropriate that in patients with liver disease, doses of verapamil should be decreased by degree of hepatic failure.

Preparation and Characterization of Solid Dispersions of Itraconazole by using Aerosol Solvent Extraction System for Improvement in Drug Solubility and Bioavailability

  • Lee, Si-Beum;Nam, Kyung-Wan;Kim, Min-Soo;Jun, Seoung-Wook;Park, Jeong-Sook;Woo, Jong-Soo;Hwang, Sung-Joo
    • Archives of Pharmacal Research
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    • v.28 no.7
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    • pp.866-874
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    • 2005
  • The objective of this study was to elucidate the feasibility to improve the solubility and bioavailability of poorly water-soluble itraconazole via solid dispersions by using supercritical fluid (SCF). Solid dispersions of itraconazole with hydrophilic polymer, HPMC 2910, were prepared by the aerosol solvent extraction system (ASES) under different process conditions of temperature/pressure. The particle size of solid dispersions ranged from 100 to 500 nm. The equilibrium solubility increased with decrease (15 to 10 MPa) in pressure and increase (40 to $60^{\circ}C$) in temperature. The solid dispersions prepared at $60^{\circ}C$/15 MPa showed a slight increase in equilibrium solubility (approximately 27-fold increase) when compared to pure itraconazole, while those prepared at $60^{\circ}C$/10MPa showed approximately 610-fold increase and no endothermic peaks corresponding to pure itraconazole were observed, indicating that itraconazole might be molecularly dispersed in HPMC 2910 in the amorphous form. The amorphous state of itraconazole was confirmed by DSC/XRD data. The pharmacokinetic parameters of the ASES-processed solid dispersions, such as $T_{max},\;C_{max},\;and\;AUC_{0-24h}$ were almost similar to $Sporanox_{\circledR}$ capsule which shows high bioavailability. Hence, it was concluded that the ASES process could be a promising technique to reduce particle size and/or prepare amorphous solid dispersion of drugs in order to improve the solubility and bioavailability of poorly water-soluble drugs.

HPLC Determination and Steady-State Bioavailability Study of Levodropropizine Sustained-release Tablets in Dogs

  • Yan, Lin;Li, Tongling;Zhang, Rongqin;Xu, Xiaohong;Zheng, Pengcheng
    • Archives of Pharmacal Research
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    • v.29 no.6
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    • pp.514-519
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    • 2006
  • A simple HPLC method using UV detection was developed and validated for the determination of levodropropizine (LDP) In dog plasma. The sample was prepared for injection using a liquid-liquid extraction method with 1-phenypiperazine as the internal standard. The mobile phase was methanol - diethylamine solution (0.05 M) (20:80, v/v, pH adjusted to 3.0 with $H_3PO_4$) with a detection wavelength of 240 nm. The limit of quantitation (LOQ) of LDP in a biological matrix was determined to be 25.25 ng/mL. The calibration curve was linear across the concentration range of 25.25 to 2020 ng/mL. The intra-day and inter-day precision values (CV%) were within 7% and accuracy (R.E. %) was within 6% of the nominal values for medium (252.5 ng/mL) and high (2020 ng/mL) LDP concentrations. For the LDP concentration at the LOQ, the intra-day and inter-day precision and accuracy were within 20% and 10%, respectively. The average absolute recovery for LDP was 70.28%. This method was successfully used to analyze plasma samples in a steady-state bioavailability study of a newly developed sustained-release LDP tablets (SR) using immediate-release tablets (IR) as the reference. The relative bioavailability of the SR was determined to be $106.3\;{\pm}\;12.8%$ (n=6). The $C_{max}$ of the SR was significantly lower (p<0.05), and the $t_{max}$ was significantly longer than that of the IR (p<0.05). The results of ANOVA and two one-sided tests indicated that the SR exhibited acceptable sustained release properties and was bioequivalent to the IR.

Self-Reprogramming of Spermatogonial Stem Cells into Pluripotent Stem Cells without Microenvironment of Feeder Cells

  • Lee, Seung-Won;Wu, Guangming;Choi, Na Young;Lee, Hye Jeong;Bang, Jin Seok;Lee, Yukyeong;Lee, Minseong;Ko, Kisung;Scholer, Hans R.;Ko, Kinarm
    • Molecules and Cells
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    • v.41 no.7
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    • pp.631-638
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    • 2018
  • Spermatogonial stem cells (SSCs) derived from mouse testis are unipotent in regard of spermatogenesis. Our previous study demonstrated that SSCs can be fully reprogrammed into pluripotent stem cells, so called germline-derived pluripotent stem cells (gPS cells), on feeder cells (mouse embryonic fibroblasts), which supports SSC proliferation and induction of pluripotency. Because of an uncontrollable microenvironment caused by interactions with feeder cells, feeder-based SSC reprogramming is not suitable for elucidation of the self-reprogramming mechanism by which SSCs are converted into pluripotent stem cells. Recently, we have established a Matrigel-based SSC expansion culture system that allows longterm SSC proliferation without mouse embryonic fibroblast support. In this study, we developed a new feeder-free SSC self-reprogramming protocol based on the Matrigel-based culture system. The gPS cells generated using a feeder-free reprogramming system showed pluripotency at the molecular and cellular levels. The differentiation potential of gPS cells was confirmed in vitro and in vivo. Our study shows for the first time that the induction of SSC pluripotency can be achieved without feeder cells. The newly developed feeder-free self-reprogramming system could be a useful tool to reveal the mechanism by which unipotent cells are self-reprogrammed into pluripotent stem cells.

Theoretical Study for Thermally Activated Delayed Fluorescence (TADF) Property in Organic Light-Emitting Diode (OLED) Candidates (유기발광소재(OLED) 후보물질의 지연형광(TADF) 성질에 대한 이론적 연구)

  • Seo, Hyun-il;Jeong, Hyeon Jin;Yoon, Byung Jin;Kim, Seung-Joon
    • Journal of the Korean Chemical Society
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    • v.63 no.3
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    • pp.151-159
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    • 2019
  • The TADF properties for carbazol-dicyanobenzene, carbazol-diphenyl sulfone, carbazol-benzonitrile derivatives as OLED candidate materials are theoretically investigated using density functional theory (DFT) with $6-31G^{**}$, cc-pVDZ, and cc-pVTZ basis sets. The optimized geometries, harmonic vibrational frequencies, and HOMO-LUMO energy separations are predicted at the B3LYP/$6-31G^{**}$ level of theory. The harmonic vibrational frequencies of the molecules considered in this study show all real numbers implying true minima. The time dependent density functional theory (TD-DFT) calculations have been also applied to investigate the absorption and emission wavelength (${\lambda}_{max}$), energy differences (${\Delta}E_{ST}$) between excited singlet ($S_1$) and triplet ($T_1$) states of candidate materials.