• Mass Spectrometry Letters
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• v.10 no.2
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• pp.66-70
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• 2019

Rivaroxaban (RRN) is the first available active direct factor Xa inhibitor (anticoagulant) with oral administration. Due to its success in market, there have been efforts to develop various RRN formulations, and the development of good analytical methods for its in vivo evaluation is an essential prerequisite. Thus, here, a simple and efficient method to determine RRN in rat plasma using liquid-liquid extraction (LLE) and liquid chromatography and multiple reaction monitoring (LC-MRM) was presented. The use of ethyl acetate as the LLE solvent results appropriate extraction and purification of RRN and it also helps the significant reduction of rat plasma volume required for RRN quantitation. The developed method showed good analytical performance including specificity, linearity ($r^2{\geq}0.999$ within 0.5 - 500 ng/mL), sensitivity (the lower limit of quantitation at 0.5 ng/mL), accuracy (89.3 - 107.0%), precision (${\geq}12.7%$), and recovery (89.2 - 105.7%). Additionally, RRN in sample extracts showed good stability. Finally, the applicability of the validated method to the PK evaluation of RRN was confirmed after its oral administration to normal rats. The present method is the first analytical method employing LLE for the simple and efficient extraction and purification of RRN in rat plasma. Therefore, the present method can contribute to the development of new RRN formulations as well as to the monitoring of RRN in special clinical situations through its efficient determination in various samples with or without minor modification.

• Journal of Microbiology
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• v.45 no.6
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• pp.593-596
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• 2007

In this study, we describe the development of a simple and efficient method for cell lysis via the insertion of a bacteriophage lambda lysis gene cluster into the pET22b expression vector in the following order; the T7 promoter, a gene for a target protein intended for production, Sam7 and R. This insertion of R and Sam7 into pET22b exerted no detrimental effects on cellular growth or the production of a target protein. The induction of the T7 promoter did not in itself result in the autolysis of cells in culture but the harvested cells were readily broken by freezing and thawing. We compared the efficiency of the cell lysis technique by freezing and thawing to that observed with sonication, and determined that both methods completely disintegrated the cells and released proteins into the solution. With our modification of pET22b, the lysis of cells became quite simple, efficient, and reliable. This strategy may prove useful for a broad variety of applications, particularly in experiments requiring extensive cell breakage, including library screening and culture condition exploration, in addition to protein purification.

• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.2
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• pp.82-86
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• 1998

The maltose binding protein (MBP) fusion protein system is a versatile tool to express and isolate recombinant proteins in E. coli. In this system, MBP fusion proteins are efficiently isolated from whole cell lysate using amylose conjugated agarose beads and then eluted by competition with free maltose. Since MBP is a rather large molecule (∼42 kDa), for further experiments, the MBP part is usually proteolytically cleaved from the fusion protein and subsequently removed by ion-exchange chromatography or rebinding to amylose columns after washing out excess and MBP-bound maltose. In the present study, we have developed an improved method for the removal of cleaved MBP, which is advantageous over conventional methods. In this method, factor Xa cleaved MBP fusion proteins were incubated with Sepharose beads conjugated with MBP specific monoclonal antibodies and then precipitated buy centrifugation, resulting in highly purified proteins in the supernatant.

• Archives of Plastic Surgery
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• v.33 no.1
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• pp.21-30
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• 2006

Alginate, a polymer of guluronic and mannuronic acid, is used as a scaffolding material in biomedical applications. The research was to produce highly-purified alginate from seaweeds and to evaluate the efficacy of alginate as dermal substrate. Our alginate purification method showed a production rate as high as 25%. The purified alginate contained little polyphenol contents and endotoxin, proteins. For study of wound healing, full thickness skin defects were made on the dorsal area of the animal models. And then alginate, fibroblast-growth-factor mixed alginate, alginate-collagen complex, vaseline gauze as control were applied on the wound, respectively, and were evaluated grossly and histopathologically. For biocompatibility test, alginate and alginate-collagen complex discs were implanted on the back of Sprague-Dawly rats. Four weeks after implantation, the animals were examined immunologically against alginate and collagen. Alginate and FGF-mixed alginate, alginate-collagen complex group showed statistically higher percentage of wound contraction and wound healing than control group(p<0.05). Alginate-collagen complex group and FGF-mixed alginate group showed statistically higher percentage of wound healing than alginate group. The experiment of biocompatibility and immunologic reaction against impanted alginate or collagen needs more investigation. Highly-purified alginate from seaweeds by our purification method, showed the effect of wound healing, and addition of FGF or collagen increases the alginate's wound healing effect. It shows the possibility of alginate as a dermal substrate.

• Journal of Life Science
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• v.6 no.1
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• pp.14-26
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• 1996

In order to effectively utilize fish(Cod, Gadus macrocephalus) bone obtained as fish waste in fish manufactory, the preparation of the fish bone gelatin were attempted by heat extracting method from collagen protein contained in the fish bone. The methods of two kinds pretreatments (the B-type by alkali pretreatment and the E-type by enzyme pretreatment) for fish bone and the optimal extraction conditions to prepare gelatin from pretreated fish bone were investigated. Physical properties and functionalities of the two type fish bone gelatins obtained were compared with the commercial gelatin and the fish skin gelatin. The optimal extraction conditions of the B-type and the E-type gelatins were 5 folds of added water with material(w/w), pH 5.0, 3 hrs of extraction time and 60$\circ$C of extraction temperature. The yield of the B-type and the E-type gelatins were 32.6% and 28.1 %, respectively. The B-type gelatin was superior to the E-type un all physical properties. Molecular weight of the B-type was larger than that of the E-type due to its pretreatment method. Among the composition of amino acids, the amino acids such as glycine, alanine, glutamic acid and imino acids(proline and hydroxyproline) were responsible for 68$\sim$70% of the total amino acids. Functionalities of the fish bone gelatin were almost similar to commercial gelatin.

• International Journal of Automotive Technology
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• v.7 no.3
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• pp.277-282
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• 2006

With the aim of achieving half the regulated value of EURO-3 Emission Regulations, an ultra low emission motorcycle has been developed based on a motorcycle with an 1800 $cm^3$, horizontal opposed 6-cylinder engine. For the fuel supply system, an electronically controlled fuel injection system was applied. For the emission purification system, three-way catalysts, a feedback control system with a LAF(Linear Air-Fuel ratio) sensor, and a secondary air induction system were applied. To reduce CO and HC emissions during cold starting, an early catalyst activation method combining RACV(Rotary Air Control Valve) and retarded ignition timing was applied. After the catalyst activation, air-fuel ratio was controlled to maximize the purification ratio of the catalyst according to vehicle speed. For the air-fuel ratio control system, the LAF sensor was used. Furthermore, fine adjustment by the LAF feedback control reduced torque fluctuation due to the air-fuel ratio change. As a result, smooth ride feeling was maintained. Owing to these technologies, half the regulated value of EURO-3 has been achieved without any negative impact to the large-scaled motorcycles' drivability. This paper presents the developed ultra low emission technologies including the control method using an LAF sensor.

• Analytical Science and Technology
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• v.24 no.6
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• pp.477-483
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• 2011

This work is on the synthesis of EMI-$BF_4$ (1-ethyl-3-methylimidazolium tetrafluoroborate) and purification of spectroscopic grade using aluminium oxide method, activated charcoal method, and liquid/liquid fractional extraction method in order to make supercapacitor finally. But the aluminum oxide method and the activated charcoal method were not suitable for obtaining high-purity ionic liquids. The liquid/liquid fractional distillation method turned out that as the concentration of solvent ($H_2O$) was increased, the higher purity of EMI-$BF_4$ was obtained and the electrical capacity of this compound was increased to higher value. When the solvent was changed to from methylene chloride to 1,2-dichloroethane, the higher purity of EMI-$BF_4$ was obtained.

• KSBB Journal
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• v.14 no.1
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• pp.9-16
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• 1999

Transforming growth factor $\beta$1(TGF-$\beta$1) has potentials to be used as a new therapeutic agent. However, studies with TGF-$\beta$ were hindered by its high cost. In this study, we developed an improved method to purify TGF-$\beta$1 from human platelets, for which four purification steps were used: platelet extraction, gel filtration, cation exchange chromatography, and reverse phase high performance liquid chromatography. After a final step of purification, a pure protein with a molecular weight of 25,000 corresponding to the commercially available TGF-$\beta$1 was obtained, which were confirmed by silver staining and Western blotting after SDS-polyacrylamide gel electrophoresis (SDS-PAGE). It was confirmed by the inhibitory effects of TGF-$\beta$1 on a mink lung epithelial cell line that the purified TGF-$\beta$1 had its biological activity, whose activity is slightly higher than that of the commercially available TGF-$\beta$1. About 3.7$\mug of the purified TGF-$\beta$1 was obtained from 10 units of concentrated human platelets, the final yield of which is about 21%.

• Journal of Radiation Protection and Research
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• v.24 no.2
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• pp.73-86
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• 1999

The ion-exchange method for the purification of primary coolant has been used broadly in PWR(pressurized water reactor)-type nuclear power plants due to its high decontamination efficiency, simple system, and easy operation. However, its non-selective removal of metal and non-radionuclides shortens its life, resulting in the generation of a large amount of waste ion-exchange resin. In this study, the feasibility of electrodeionization (EDI) was investigated for the purification of primary cooling water using synthetic solutions under various experimental conditions as an alternative method for the ion exchange. The results shows that as the feed flow-rate increased, the removal efficiency increased and the power consumption decreased. The removal rate was observed as a 1000 decontamination factor(DF) at a nearly constant level. For the synthetic solution of 3 ppm TDS (Total Dissolved Solid), the power consumption was 40.3 mWh/L at 2.0 L/min of feed flow rate. The higher removal rate of metal species and lower power consumption were obtained with greater resin volume per diluting compartment. However, the flow rate of the EDI process decreased with the elapsed time because of the hydrodynamic resistivity of resin itself and resin fouling by suspended solids. Thus, the ion-exchange resin was replaced by an ion-conducting spacer in order to overcome the drawback. The system equipped with the ion-conducting spacer resolved the problem of the decreasing flow rate but showed a lower efficiency in terms of the power consumption, the removal rate of metal species and current efficiency. In the repeated batch operation, it was found that the removal efficiency of metal species was stably maintained at DF 1000.

• Journal of The Korean Society of Agricultural Engineers
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• v.62 no.5
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• pp.39-46
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• 2020

This study evaluated the nitrogen oxide (NO_x) removal efficiency by air purification concrete blocks with titanium dioxide (TiO₂). The concrete in the mixtures had a 30% water:cement ratio, to which TiO₂ was added at 0%, 5%, and 10% of cement weight. The compressive strength reduction rate and removal efficiency of NO_x were investigated. The result of the compressive strength test in the study indicated that addition rate of TiO₂ did not lead to signifcant effect. In terms of the average removal efficiency of NO_x, mix No. 1 using a TiO₂ mixing ratio of 0% had a removal efficiency of 0.57% on average; thus, the removal effect w as not significant. For the other samples prepared by mixing, the average removal efficiencies for mix No. 2 (5% TiO₂) were 58.86% and 62.05% for normal and washing surface treatments, respectively, and those of sample No. 3 (10% TiO₂) were 59.94% and 67.61%. mixs No. 4 (5%) and No. 5 (10%), in which TiO₂ diluted with distilled water was sprayed onto the block surface, had an average NO_x removal efficiency of 61.72% and 68.48%, respectively. In terms of NOx removal efficiency, Mixs No. 3 and No. 5 with 10% TiO₂ were better than Mixs No. 2 and No. 4 with 5% TiO₂. In addition, analyzing the NO_x removal efficiency results from the fixing method, it was capable to apply mixing (washing) and the diluted spray methods. Therefore, it was found that the diluted spray method applied in this study can be employed in any manufacture of air purification concrete blocks.