• Title/Summary/Keyword: Half-life(t1/2)

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Engineering CotA Laccase for Acidic pH Stability Using Bacillus subtilis Spore Display

  • Sheng, Silu;Jia, Han;Topiol, Sidney;Farinas, Edgardo T.
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.507-513
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    • 2017
  • Bacillus subtilis spores can be used for protein display to engineer protein properties. This method overcomes viability and protein-folding concerns associated with traditional protein display methods. Spores remain viable under extreme conditions and the genotype/phenotype connection remains intact. In addition, the natural sporulation process eliminates protein-folding concerns that are coupled to the target protein traveling through cell membranes. Furthermore, ATP-dependent chaperones are present to assist in protein folding. CotA was optimized as a whole-cell biocatalyst immobilized in an inert matrix of the spore. In general, proteins that are immobilized have advantages in biocatalysis. For example, the protein can be easily removed from the reaction and it is more stable. The aim is to improve the pH stability using spore display. The maximum activity of CotA is between pH 4 and 5 for the substrate ABTS (ABTS = diammonium 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate). However, the activity dramatically decreases at pH 4. The activity is not significantly altered at pH 5. A library of approximately 3,000 clones was screened. A E498G variant was identified to have a half-life of inactivation ($t_{1/2}$) at pH 4 that was 24.8 times greater compared with wt-CotA. In a previous investigation, a CotA library was screened for organic solvent resistance and a T480A mutant was found. Consequently, T480A/E498G-CotA was constructed and the $t_{1/2}$ was 62.1 times greater than wt-CotA. Finally, E498G-CotA and T480A/E498G-CotA yielded 3.7- and 5.3-fold more product than did wt-CotA after recycling the biocatalyst seven times over 42 h.

Effect of Thidiazuron on the Formation of Micro-tubers and Plantlet Regeneration of Pinellia ternata T. (Thidiazuron이 반하(半夏)의 자구(子球) 및 식물체 재생에 미치는 영향)

  • Kim, Jae-Kwang;Cho, Hye-Kyoung;Yu, Chang-Yeon;Ahn, Sang-Deuk;Lim, Hak-Tae
    • Korean Journal of Medicinal Crop Science
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    • v.5 no.1
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    • pp.21-27
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    • 1997
  • This experiments were conducted to determine the effect of thidiazuron on forming tuberlets and plant regeneration of Pinellia ternata T. by tissue culture. The addition of $5\;{\mu}M$ TDZ to the medium had better regeneration than that of any other treatments of NAA and TDZ. At the combination treatments of NAA and TDZ, as the level of thidiazuron increased, the rate of shoot regeneration was incresed while the increment of NAA concentration inhibited the rate of shoot regeneration. The supplement of $5\;{\mu}M$ thidiazuron produced the best number of micro-tubers per explant and the number of micro-tuber formed was 25 in MS medium and 29 in MG medium on 30 day culture, respectively. Microtuber formation was the best on MG medium with 1.0 mg/l NAA and $5\;{\mu}M$ thidiazuron. MG medium was superior to MS and B5 medium for the growth of tuberlets. Half strength of MS medium with NAA 2 mg/l was the most effective for root formation. Rooting ability on nursery soil of plantlets produced in in vjtro was good as a 80% after 3 weeks.

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A Study about the Experienced Problems and the Demands of Counseling of the Adolescents according to the Physical, Psychological, Sexual and the Socio-cultural Problem Categories (일부 지역 청소년들의 신체적$\cdot$정신적$\cdot$성적$\cdot$사회문화적 영역별 문제경험정도와 상담의 필요성에 관한 연구)

  • Kim Jin Kyung;Kim Eun Sook;Lee Jung Ae
    • Journal of Korean Public Health Nursing
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    • v.11 no.2
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    • pp.152-170
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    • 1997
  • The purpose of this study was to identify physical. psychological. sexual and socio-cultural problems that the adolescents have experienced and to assess the demands of counseling about the experienced problems. The design of this study was descriptive-survery study. The sample size was 601 middle and high school students. Data analysis was done by frequencies. Percent. t -test. ANOVA. Pearson correlation coefficiency, stepwise regression and Cronbach's $\alpha$ Score was produced for the reliability of the tool by using the item analysis method. The results was as follows : 1. The distribution of the each item that the adolescents experienced according to the physical, psychalogical. Sexual and Socio-cultural problem Categories was examined. In the physical problem category, the number of items that half and more than half of the adolescents experienced of often or frequently were 10 among all 19 items. In the psychological problem category. 13 item were among all 19 items. In the sexual problem category, 1 item among all 17 items and in the socio-cultural problem category, 10 items were among all 30 items. 2. The distribution of the each items that the adolescents wanted to the counseling according to the physical. psychological. sexual and socio-cultural problem categories was examined. In the physical problem category, the numbers of the items that half and more than half of the adolescents wanted counseling were 1 among all 19 items. In the psychological problem category, the numbers of the items that half and more than half of the adolescents wanted counseling were 2 items among all 19 items. In the sexual problem category, 0 item among all 17 items and in the socio-cultural problem category, 2 items were among all 30 items. 3. In the comparison of the female student and male student about the experienced problems and the demands of counseling according to the physical. psychological. sexual and socio-cultural problem categories. There was a significant difference statistically in all categories between groups(P<.001) 4. In the comparison of high school students and middle school students about the demands of counseling according to the physical. psychological. sexual and socio-cultural problem categories(P<.001). Particulary, grade II of high school students have experienced problems according to the physical. psychological. sexual and socio-cultural problem categories. There was a significant difference statistically in all categories(P<.001). 5. In the correlation between the experienced problem and the demands of counseling according to 4 categories. there was the positive correlation at P<.001 level. 6. The less the satisfaction for family and school life. the more experienced problems according to the physical. psychological. sexual and socio-cultural problem categories. There was a significant difference statistically in all categories(P<.001).

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Characterization of Laccase Purified from Korean Trametes hirsuta S1 (한국산 흰구름버섯(Trametes hirsuta S1)으로부터 정제된 Laccase의 특성)

  • Lim, Hyung-Seon;Yoon, Kyung-Ha
    • The Korean Journal of Mycology
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    • v.32 no.2
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    • pp.112-118
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    • 2004
  • Laccase produced by Trametes hirsuta S1 isolated from Korea was partially purified and characterized using ultrafiltration, anion exchange chromatography and affinity chromatography. The laccase was produced as the predominant extracellular enzyme during primary metabolism. Neither lignin peroxidase nor veratryl alcohol oxidase (VAO) were detected in the culture fluid. Addition of 2,5-xylidine enhanced 4-fold laccase production. Purified laccase was a single polypeptide having a molecular mass of approximately 66 kDa, as determined by SDS-polyacrylamide gel electrophoresis, and carbohydrate content of 12%. $K_{m}\;and\;V_{max}$ values for laccase with ABTS [2,2-azinobis (3-ethylbenzthiazoline 6-sulfonic acid)] as a substrate (Lineweaver-Burk plot) was determined to $51.2\;{\mu}M\;and\;56.8\;{\mu}mole$, respectively. The optimal pH for laccase activity was found to be 3.0. The enzyme was very stable for 1 hour at $50^{\circ}C$. Half-life ($t_{1/2}$) of the enzyme was about 20 min at $70^{\circ}C$. Spectroscopic analysis of purified enzyme indicated that the enzyme was typical of copper-containing protein. Substrate specificity and inhibitor studies for laccase also indicated to be a typical fungal laccase. The N-terminal amino acid sequence of the T. hirsuta S1 laccase showed 100% of homology to those of laccase from C. hirsutus.

Bioequivalence of Ramiprin® tablet to Tritace Protect® tablet (Ramipril 10 mg) (트리테이스프로텍트 ®정(라미프릴 10mg)에 대한 라미프린 ®정의 생물학적동등성)

  • Oh, Soo-Yeon;Cho, Jong-Tae;Kim, Hyung-Gun;Kim, Yoon-Gyoon
    • Journal of Pharmaceutical Investigation
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    • v.38 no.1
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    • pp.73-78
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    • 2008
  • To evaluate the bioequivalence of two ramipril formulations, a standard 2-way randomized cross-over study was conducted in twenty-six healthy male Korean volunteers. A single oral dose of 10 mg of test formulation $Ramiprin^{(R)}$ (tablet) or reference formulation Tritace $Protect^{(R)}$ (tablet) was administered with one-week washout period. Plasma concentrations of ramipril were assayed over a period of 12 hr with a well validated method using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The values of area under the plasma concentration-time curve, from time zero to last sampling time $(AUC_t)$ and from time zero to time infinity $(AUC_{inf})$ were $77.45{\pm}44.78\;and\;78.96{\pm}45.64$ for test, and $70.30{\pm}42.27\;and\;71.99{\pm}43.55ng\;hr/mL$ for reference formulation, respectively. Similarly, maximum concentration $(C_{max})$ and elimination half-life $(t_{1/2})$ were $65.61{\pm}19.96ng/mL$ and $2.15{\pm}0.75hr$ for test, and $63.63{\pm}25.50ng/mL$ and $2.16{\pm}0.73hr$ for reference formulations, respectively. Time to reach maximum concentration $(T_{max})$ for the test and the reference, were $0.51{\pm}0.22hr\;and\;0.52{\pm}0.18hr$, respectively. The parametric 90% confidence intervals on the mean of the differences between the two formulations (test-reference) of the log-transformed values of $AUC_t\;and\;C_{max}$ were 1.03 to 1.19 and 0.98 to 1.17, respectively. The overall results indicate that the two formulations are bioequivalent and can be prescribed interchangeably.

The Properties of the Metal Hydride electrodes prepared by Silicon Sealant (Si-sealant를 이용하여 제조한 금속수소화물 전극의 특성)

  • CHOI, Jeon;PARK, Choong-Nyeon
    • Transactions of the Korean hydrogen and new energy society
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    • v.4 no.2
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    • pp.23-28
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    • 1993
  • The $(LM)Ni_{4.5}Co_{0.1}Mn_{0.2}Al_{0.2}$ hydrogen storage alloy powders were conducted 25wt% electroless copper plating in an acidic bath. For the preparation of a hydride electrodes, the copper coated alloy powder was mixed with Si-sealant(organosilicon) and compacted with $6t/cm^2$ at room temperature. The electrode characteristics were examined through electrochemical measurements in a half cell. As a sealant contents increased, the initial discharge capacity of si-sealant bounded electrode was lower and the activation rate in high current density was slower. For extended cycles, however, the electrodes with the Si-sealant were superior in a high rate discharge and useful range of temperature over the sealant-free electrode. In addition, the cycle life increased with increasing the amount of Si-sealant added. It can be suggested from the results that the Si-sealant as a binder could be applied to the preparation of the metal hydride electrode.

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Pharmacokinetic Study of Florfenicol in Healthy and Vibriosis-infected Pseudosciaena crocea after Oral Administration

  • Wang, Li;Han, Yan-nan;Jin, Shan;Ma, Yin;Wang, Guo-liang;Zhao, Qing-song;Chen, Yin-er
    • Journal of Applied Biological Chemistry
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    • v.58 no.4
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    • pp.363-368
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    • 2015
  • The pharmacokinetics of florfenicol were studied in healthy and vibriosis-infected large yellow croaker (Pseudosciaena crocea) following administration of a single oral dose of $20mg{\cdot}kg^{-1}$ at $25{\pm}2^{\circ}C$. After oral administration, florfenicol levels in tissues (liver, kidney, muscle, serum, and skin) were analyzed using high-performance liquid chromatography. A two-compartment open model was used to describe the pharmacokinetics of florfenicol following oral administration. Compared to the healthy group, the absorption rate of vibriosis-infected fish significantly decreased, peak-time ($T_{max}$) delayed, maximum concentration ($C_{max}$) declined, total body clearance decreased, the elimination half-life ($T_{1/2{\beta}}$) was extended, and the area under the curve increased. These results indicate that a $20mg{\cdot}kg^{-1}$ oral dose of florfenicol administered once daily continuously for 4 or 5 days can be used for the treatment of Vibrio alginolyticus infection in large yellow croaker (Pseudosciaena crocea).

Effect of Morin on the Pharmacokinetics of Nifedipine in Rats (흰쥐에서 모린이 니페디핀의 약물동태에 미치는 영향)

  • Lee, Chong-Ki;Choi, Jun-Shik
    • YAKHAK HOEJI
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    • v.51 no.3
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    • pp.169-173
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    • 2007
  • The aim of this study was to investigate the effect of morin on the pharmacokinetics of nifedipine in rats. The pharmacokinetic parameters of nifedipine were measured after the oral administration of nifedipine (5 mg/kg) in the presence or absence of morin (1.5, 7.5 and 15 mg/kg, respectively). Compared to the control groups, the presence of 7.5 mg/kg and 15 mg/kg of morin significantly (p<0.05) increased the area under the plasma concentration-time curve (AUC) of nifedipine by 48.5${\sim}$68.2%, and the peak concentration (C$_{max}$,) of nifedipine by 59.9~84.2%. The absolute bioavailability(AB%) of nifedipine was significantly (p<0.05) increased by 21.5${\sim}$24.5% compared to the control (14.5%). While there was no significant change in the time to reach the peak plasma concentration (T$_{max}$) and the terminal half-life (T$_{1/2}$) of nifedipine in the presence of morin. It might be suggested that morin altered disposition of nifedipine by inhibition of both the first-pass metabolism and p-glycoprotein (P-gp) efflux pump in the small intestine of rats. In conclusion, the presence of morin significantly enhanced the oral bioavailability of nifedipine, suggesting that concurrent use of morin or morin-containing dietary supplement with nifedipine should require close monitoring for potential drug interaction.

Residue level and pharmacokinetics of trichlorfon in the Japanese eel (Anguilla japonica) after bath treatment (Trichlorfon (TCF)의 약욕 투여에 따른 뱀장어 체내 약물 잔류량 및 약물동태학 연구)

  • Jo, Hyun Ho;Chung, Joon Ki
    • Journal of fish pathology
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    • v.35 no.1
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    • pp.93-102
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    • 2022
  • This study performed a trichlorfon (TCF) residue and pharmacokinetic analysis with Japanese eels, Anguilla japonica, to obtain baseline data to establish the maximum residue level (MRL) of TCF in A. japonica. After dipping A. japonica in 30 ppm and 150 ppm of TCF at 28℃ and 18℃, drug residue in the body was analyzed with LC-MS/MS, and these results were further analyzed with the PK solver program to obtain the pharmacokinetic parameters of TCF in the serum, muscles, and liver. The maximum concentrations (Cmax) in the serum, muscles, and liver were 25.87-357.42, 129.91-1043.73, and 40.47-375.20, respectively, and the time to maximum concentration (Tmax) was 0.13-1.32h, 1.17-3.34h, and 0.14-5.40h, respectively. The terminal elimination half-life (T1/2) was 2.13-3.92h, 5.30-10.35h, and 0.65-13.81h, respectively. In the 30 mg/L concentration group, TCF was not detected in the serum of eels 96 hours after bathing, and was below the detection limit after 336 hours in muscle and liver. On the other hand, in the 150 mg/L concentration group, TCF was not detected in the serum of eels 336 hours after bathing, but was detected in muscle and liver at 336 hours. In conclusion, the results of this study would be useful in establishing the MRL of TCF in farmed A. japonica.

Study on Physicochemical Properties of Pesticide. (I) Water Solubility, Hydrolysis, Vapor Pressure, and n-Octanol/water Partition Coefficient of Captafol (농약의 물리화학적 특성연구 (I) Captafol의 수용성, 가수분해, 증기압, 옥탄올/물 분배계수)

  • Kim, Jeong-Han;Lee, Sung-Kyu;Kim, Yong-Hwa;Kim, Kyun
    • Applied Biological Chemistry
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    • v.40 no.1
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    • pp.71-75
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    • 1997
  • Important physicochemical properties of captafol [N-(1,1,2,2-tetrachloro-ethylthio)cyclohex-4-ene-1,2-dicarboximide], water solubility, vapor pressure, hydrolysis and octanol/water partition coefficient(Kow) were measured based on the standard EPA and OECD methods. Water solubility of the chemical was 2.24 ppm at $25^{\circ}C$. Half-life by hydrolysis at $25^{\circ}C$ in the buffer solution of pH 3.0, pH 7.0, and pH 8.0 was 77.8 hr, 6.54 hr and 0.72 hr, respectively, demonstrating instability in alkaline solution. The half-life in acid condition was not significantly different by temperature change, however, that in neutral or alkaline solution became shorter at $40^{\circ}C$. Hydrolysis study with a reference compound, diazinon, proved that the experimental method of the present study is reliable. Vapor pressure of captafol, $8.27{\times}10^{-9}$ torr at $20^{\circ}C$, was calculated from the equation, log P=6.94-(4401.6/T) plotted on the experiment results under different temperature conditions, 40, 50, and $60^{\circ}C$. pressure of captafol, the contamination of captafol would not happen easily in environment by vaporization. High Kow value of 1,523 was observed and this might result in bioconcentration through food chain when captafol was exposed. However, affecting human health through aquatic bioaccumulation is not likely to occur due to its rapid hydrolysis in the environment.

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