• Title/Summary/Keyword: time to $V_{max}$

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Characterization of a Lichenase Isolated from Soil Metagenome

  • Kim, Sang-Yoon;Oh, Doo-Byoung;Kwon, Ohsuk
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1699-1706
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    • 2014
  • A lichenase gene (mt-lic) was identified for the first time through function-based screening of a soil metagenomic library. Its deduced amino acid sequence exhibited a high degree of homology with endo-${\beta}$-1,3-1,4-glucanase (having both lichenase and chitosanase activities), encoded by the bgc gene of Bacillus circulans WL-12. The recombinant lichenase overexpressed and purified from Escherichia coli was able to efficiently hydrolyze both barley ${\beta}$-glucan and lichenan. The enzyme showed maximal activity at a pH of 6.0 at $50^{\circ}C$, with Azo-barley-glucan as the substrate. The metal ions $Mn^{2+}$, $Mg^{2+}$, $Ca^{2+}$, and $Fe^{2+}$ enhanced the enzymatic activity, whereas the $Cu^{2+}$ and $Zn^{2+}$ ions inhibited the enzymatic activity. The $K_m$ and $V_{max}$ values of the purified lichenase were determined to be 0.45 mg/ml and 24.83 U/min/mg of protein, respectively.

Optimization of $\beta$-Galactosidase Production in Stirred Tank Bioreactor Using Kluyveromyces lactis NRRL Y-8279

  • Dagbagh, Seval;Goksungur, Yekta
    • Food Science and Biotechnology
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    • v.18 no.6
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    • pp.1342-1350
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    • 2009
  • This paper investigates the production and optimization of $\beta$-galactosidase enzyme using synthetic medium by Kluyveromyces lactis NRRL Y-8279 in stirred tank bioreactor. Response surface methodology was used to investigate the effects of fermentation parameters on $\beta$-galactosidase enzyme production. Maximum specific enzyme activity of 4,622.7 U/g was obtained at the optimum levels of process variables (aeration rate 2.21 vvm, agitation speed 173.4 rpm, initial sugar concentration 33.8 g/L, incubation time 24.0 hr). The optimum temperature and pH of the $\beta$-galactosidase enzyme produced under optimized conditions were $37^{\circ}C$ and pH 7.0, respectively. The enzyme was stable over a pH range of 6.0-7.5 and a temperature range of $25-37^{\circ}C$. The $K_m$ and $V_{max}$ values for O-nitrophenol-$\beta$-D-galactopyranoside (ONPG) were 1.20 mM and $1,000\;{\mu}mol/min{\cdot}mg$ protein, respectively. The response surface methodology was found to be useful in optimizing and determining the interactions among process variables in $\beta$-galactosidase enzyme production. Hence, this study fulfills the lack of using mathematical and statistical techniques in optimizing the $\beta$-galactosidase enzyme production in stirred tank bioreactor.

Pharmacokinetic Drug Interaction between Carvedilol and Ticlopidine in Rats

  • Choi, Jun-Shik;Choi, Dong-Hyun
    • Biomolecules & Therapeutics
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    • v.18 no.3
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    • pp.343-349
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    • 2010
  • This study was designed to investigate the effects of ticlopidine on the pharmacokinetics of carvedilol after oral or intravenous administration of carvedilol in rats. Carvedilol was administered orally (3 mg/kg) or intravenously (1 mg/kg) without or with oral administration of ticlopidine (4, 12 mg/kg) to rats. The effects of ticlopidine on P-glycoprotein (P-gp) and cytochrome P450 (CYP) 2C9 activity were also evaluated. Ticlopidine inhibited CYP2C9 activity in a concentration-dependent manner with 50% inhibition concentration ($IC_{50}$) of $25.2\;{\mu}M$. In addition, ticlopidine could not significantly enhance the cellular accumulation of rhodamine 123 in MCF-7/ADR cells overexpressing P-gp. Compared with the control group (given carvedilol alone), the area under the plasma concentration-time curve (AUC) was significantly (12 mg/kg, p<0.05) increased by 14-41%, and the peak concentration ($C_{max}$) was significantly (12 mg/kg, p<0.05) increased by 10.7-73.3% in the presence of ticlopidine after oral administration of carvedilol. Consequently, the relative bioavailability (R.B.) of carvedilol was increased by 1.14- to 1.41-fold and the absolute bioavailability (A.B.) of carvedilol in the presence of ticlopidine was increased by 36.2-38.5%. Compared to the i.v. control, ticlopidine could not significantly change the pharmacokinetic parameters of i.v. administered carvedilol. The enhanced oral bioavailability of carvedilol may result from inhibition of CYP2C9-mediated metabolism rather than P-gpmediated efflux of carvedilol in the intestinal and/or in liver and renal eliminatin of carvedilol by ticlopidine.

TWO-COLOR VR CCD PHOTOMETRY OF THE INTERMEDIATE POLAR `RXS J062518.2+733433

  • Kim, Yong-Gi;Andronov Ivan L.;Park, Sung-Su;Chinarova Lidia L.;Baklanov Alexey V.;Jeon, Young-Beom
    • Journal of Astronomy and Space Sciences
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    • v.22 no.3
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    • pp.197-210
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    • 2005
  • Results of 7 nights of CCD VR photometry of the intermediate polar 1RXS J062518.2+733433 obtained at the Korean 1.8m telescope are reported. The corrected ephemeris for the orbital minimum is BJD (Orb.min) = 2453023.6159(42)+0.1966431(33) (E-1735). The corrected ephemeris for the spin maximum is BJD (spin max) = 2452893 78477 (10)+0.01374116815 (17) (E-15382) (cycle numbering corresponds to that of Staude et al.2003). The variations of the shape of the individual spin variations are highly correlated in V and R. The phase of the spin maximum is found to be dependent on the orbital phase. The corresponding semi-amplitude of sinusoidal variations of phase is $0.11{\pm}0.03.$ This new phenomenon is explained by the changing viewing conditions of the accreting magnetic white dwarf, and should be checked in further observations this star and for other intermediate polars. To avoid influence of this effect on the analysis of the long-term spin period variations, the runs of at least one orbital period are recommended. Results of time series analysis are presented in tables.

Comparative Study on the Transport Characteristics of Canalicular Liver Plasma Membrane Vesicles Prepared by Two Different Methods (제조 방법에 따른 간 모세담관막 소포계의 수송 특성 비교)

  • Song, Im-Sook;Chung, Suk-Jae;Shim, Chang-Koo
    • Journal of Pharmaceutical Investigation
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    • v.29 no.1
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    • pp.13-19
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    • 1999
  • Canalicular liver plasma membrane vesicles (cLPM) were prepared according to two different methods (Inoue method and Meier method), and were evaluated for their protein yield, enzyme activity and transport characteristics. No difference was found between the methods in the protein yield (i.e., $0.14{\pm}0.031$ and $0.15{\pm}0.050$ mglg liver for Inoue method and Meier method, respectively). The activity of alkaline phosphatase, a marker enzyme of canalicular membrane, was significantly (P<0.05) higher in the vesicles of Meier method $(3.52{\pm}0.91\;mmol/mg/hr)$than in the vesicles of Inoue method ($2.28{\pm}0.94$ mmol/mg/hr) indicating that more purified cLPM were obtained from Meier method compared with Inoue method. ATP-dependent vesicular uptake of taurocholate and tributylmethylammonium (TBuMA) was observed for vesicles of both methods, and the kinetic parameters responsible for the transport were similar between the vesicles of both methods (for example, $V_{max}:$ 9.72 nmol/mg protein/30sec and $K_m:$ 0.63 mM for Inoue method; $V_{max}:$ 10.1 nmol/mg protein/30sec and $K_m:$ 0.70 mM for Meier method). A pH gradient dependent counter transport of TBuMA was also observed for both vesicles with similar kinetic characteristics. Either the uptake of taurocholate in the absence of ATP or that of TBuMA in the absence of pH gradient, which may represent passive diffusion of respective compound into the vesicles, was more rapid for the vesicles of Meier method than for the vesicles of Inoue method. For example, passive diffusion rate constants $(K_d)$ for TBuMA uptake into the vesicles were 0.00030 and 0.00052\;{\mu}l/mg$ protein/min for the vesicles of Inoue method and Meier method, respectively. It may indicate that more leaky vesicles are obtained form the Meier method compared with the Inoue method. These aspects together with the time necessary to prepare the vesicles (i.e., 8 hr for Inoue method and 23 hr for Meier method) should be considered before selecting an appropriate method for the preparation of cLPM.

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LB30057, an Orally Effective Direct Thrombin Inhibitor, Prevents Arterial and Venous Thrombosis in Rats and Dogs

  • Park, Hee-Dong;Kim, Hee-Jin;Oh, Yeong-Soo;Kim, In-Chull;Kim, Yong-Zu;Koh, Hyun-Chul;Shin, In-Chul;Lee, Yong-Hee;Lee, Chang-Ho
    • Archives of Pharmacal Research
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    • v.26 no.3
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    • pp.224-231
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    • 2003
  • The anti-thrombotic effects of LB30057, a direct thrombin inhibitor, were evaluated with in vivo rat and dog thrombosis models. In rats, 1 mg/kg of LB30057 inhibited half of the clot formations in the inferior vena cava at 5 minutes after intravenous application. When measured at 2 hours after oral application, 100 mg/kg prevented approximately half of the clot formations in the inferior vena cava and 50 mg/kg prolonged the mean occlusion time from $15.6{\pm}1.3$ minutes to $47.2{\pm}8.3$ minutes in the carotid artery. In dogs, the formation of thrombus in the jugular vein was reduced to half at a dose range of 20-30 mg/kg at 6 hours after oral application. In addition, the LB30057 dosage required to reduce venous clot formation by approximately 80-90% in dogs was only about 10% of that required for the same reduction in rats. This is probably due to the variation in its time-dependent blood concentration profiles in each species; for example, the plasma half-life of LB71350 in dogs was longer than that in rats ($153.0{\pm}3.0$ vs. $129.7{\pm}12.7$ min at 30 mg/kg, i.v., respectively). AUG, $T_{max},{\;}G_{max}$, and BA in dogs were 59, 8.9, 9.17, and 13.3 times higher than those in rats at oral 30 mg/kg, respectively. Taken together, these results suggest that LB30057 administered orally is effective in the prevention of arterial and venous thrombosis in rats and dogs. It therefore represents a good lead compound for investigations to discover a new, orally available, therapeutic agent for treating thrombotic diseases.

Grazing Soybean to Increase Voluntary Cow Traffic in a Pasture-based Automatic Milking System

  • Clark, C.E.F.;Horadagoda, A.;Kerrisk, K.L.;Scott, V.;Islam, M.R.;Kaur, R.;Garcia, S.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.3
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    • pp.422-430
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    • 2014
  • Pasture-based automatic milking systems (AMS) require cow traffic to enable cows to be milked. The interval between milkings can be manipulated by strategically allocating pasture. The current experiment investigated the effect of replacing an allocation of grazed pasture with grazed soybean (Glycine max) with the hypothesis that incorporating soybean would increase voluntary cow traffic and milk production. One hundred and eighty mixed age, primiparous and multiparous Holstein-Friesian/Illawarra cows were randomly assigned to two treatment groups (n = 90/group) with a $2{\times}2$ Latin square design. Each group was either offered treatments of kikuyu grass (Pennisetum clandestinum Hoach ex Chiov.) pasture (pasture) or soybean from 0900 h to 1500 h during the experimental period which consisted of 2 periods of 3 days following 5 days of training and adaptation in each period with groups crossing over treatments after the first period. The number of cows trafficking to each treatment was similar together with milk yield (mean ${\approx}18$ L/cow/d) in this experiment. For the cows that arrived at soybean or pasture there were significant differences in their behaviour and consequently the number of cows exiting each treatment paddock. There was greater cow traffic (more cows and sooner) exiting pasture allocations. Cows that arrived at soybean stayed on the allocation for 25% more time and ate more forage (8.5 kg/cow/d/allocation) relative to pasture (4.7 kg/cow/d/allocation). Pasture cows predominantly replaced eating time with rumination. These findings suggest that replacing pasture with alternative grazeable forages provides no additional incentive to increase voluntary cow traffic to an allocation of feed in AMS. This work highlights the opportunity to increase forage intakes in AMS through the incorporation of alternative forages.

Pharmacokinetics and Lymphatic Delivery of Oligopeptide after Intramuscular Injection of Oligopeptide-bearing Liposomes to Rats (흰쥐에서 올리고펩타이드 함유 리포솜의 근육주사후 체내동태 및 임파이행)

  • Shin, Dae-Hwan;Cho, Byung-Suk;Choi, Kyu-Seok;Song, Suk-Gil;Lee, Chong-Kil;Chung, Youn-Bok
    • Journal of Pharmaceutical Investigation
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    • v.38 no.3
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    • pp.191-197
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    • 2008
  • The purpose of the present study was to examine the pharmacokinetics and lymphatic delivery of the oligopeptide, a model peptide of X antigen epitope peptides, after the intramuscular administration of the peptide-bearing liposomes in rats. $^{14}C$-labelled peptide was used as a tracer to analyze the peptide levels in plasma, bile, urine, tissue homogenates, and lymph nodes (superior cervical nodes, brachial nodes and superior mesenteric nodes). Model peptide rapidly disappeared from the plasma by 30 min (${\alpha}$ phase) after i.v. administration, which was followed by the late disappearance. The apparent plasma half-lives ($t_{1/2({\alpha}),app}$) of the peptide at the ${\alpha}$ phase when administered at a dose of 0.2-1.0 mg/kg were about 5 min. The maximum plasma concentration ($C_{max}$) was $1.52\;{\mu}g/mL$, after the i.m. administration of the peptide at a dose of 1.0 mg/kg. The bioavailability, which was calculated from the time zero to last quantitative time, of the i.m. administered peptide was over 60%. Of the various tissues tested, the peptide was mainly distributed in the kidney after the i.m. administration. The peptide levels in the kidney 3 hr after the i.m. administration were higher than those of maximum plasma concentration ($C_{max}$). The cumulative amounts of the peptide found in the urine 72 hr after the administration of 1.0 mg/kg were 2-folder higher than those in the bile, suggesting that the peptide is mostly excreted in the urine. Moreover, the concentrations of the peptide in the lymph nodes were as high as that of the plasma and the tissues. In conclusion, the peptide concentration in the lymph nodes was maintained by 24 hr after the i.m. administration of the peptide-bearing liposomes.

Effect of a New Hepatoprotective Agent, YH-439, on the Hepatobiliary Transport of Organic Cations (OCs): Selective Inhibition of Sinusoidal OCs Uptake without Influencing Glucose Uptake and Canalicular OCs Excretion

  • Hong Soon Sun;Li Hong;Choi Min Koo;Chung Suk Jae;Shim Chang Koo
    • Archives of Pharmacal Research
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    • v.28 no.3
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    • pp.330-334
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    • 2005
  • The effect of a new hepatoprotective agent, YH-439, on the hepatobiliary transport of a model organic cation (OC), TBuMA (tributylmethylammonium), was investigated. The area under the plasma concentration-time curve (AUC) from time zero to 4 h following iv administration of TBuMA (6.6 $\mu$mol/kg) was increased significantly when YH-439 in corn oil (300 mg/kg) was orally administered to rats 24 h prior to the experiment. Nevertheless, the cumulative biliary excretion of TBuMA remained unchanged. As a consequence, the apparent biliary clearance ($CL_b$) of TBuMA was decreased significantly as a result of YH-439 pretreatment, consistent with the fact that the in vivo excretion clearance of TBuMA across the canalicular membrane ($CL_{exc}$) was not changed by the pretreatment. The in vitro uptake of TBuMA into isolated hepatocytes was decreased by one half by the pretreatment, owing to a decrease in the apparent V$_{max}$ and $CL_{linear}$, but the $K_m$ for the process remained constant. Most interestingly, however, the sinusoidal uptake of glucose, a nutrient, into hepatocytes was not influenced by the pretreatment, suggesting the YH-439 pretreatment specifically impaired the sinusoidal uptake of OCs. Thus, the OC-specific inhibition of hepatic uptake, without influencing the uptake of glucose, a nutrient, appeared to be associated with the hepatoprotective activity of YH-439.

Surface Micro-Climate Analysis Based on Urban Morphological Characteristics: Temperature Deviation Estimation and Evaluation (도시의 지표형태학적 특성에 기반한 지면미기후 분석: 기온추정 및 평가)

  • Yi, Chaeyeon;An, Seung Man;Kim, KyuRang;Kwon, Hyuk-gi;Min, Jae-Sik
    • Atmosphere
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    • v.26 no.3
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    • pp.445-459
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    • 2016
  • Air temperature deviation (ATD) is one of major indicators to represent spatial distribution of urban heat island (UHI), which is induced from the urbanization. The purpose of this study is to evaluate the accuracy of air temperature deviation about Climate Analysis Seoul (CAS) workbench, which had developed by National Institute Meteorological Science and TU Berlin. Comparison and correlation analysis for CAS ATD including meso-scale air temperature deviation, local-scale air temperature deviation, total air temperature deviation, surface heat flux deviation, cold air production deviation among meso-scale numerical modelling variable in 'Seoul Region', micro-scale numerical modelling in 'Detail Region', and CAS workbench variable using observation data in ground stations. Comparison between night time OBS ATD and CAS ATD show that have most close values. Most of observations ($dT_{max}$ and $dT_{min}$) have highly positive ($dT_{SHP}$, $dT_{CA}$, MD, TD, $f_{BS}$, $f_{US}$, $f_{WS}$, $h_B$) and negative ($f_{VS}$, $f_{TV}$, $h_V$, Z) correlations. However, CAS workbench needs further improvement of both observational framework and analytical framework to resolve the problems; (1) night time OBS ATD of has closer values in compare with at high rise mountain area and (2) correlations are very dependable to meteorological scale.