• BMB Reports
• /
• v.40 no.5
• /
• pp.832-838
• /
• 2007

A novel inhibitory protein against blood coagulation factor Va (FVa) was purified from muscle protein of granulated ark (Tegillarca granosa, order Arcoida, marine bivalvia) by consecutive FPLC method using anion exchange and gel permeation chromatography. In the results of ESI-QTOF tandem mass analysis and database research, it was revealed that the purified T. granosa anticoagulant protein (TGAP) has 7.7 kDa of molecular mass and its partial sequence, HTHLQRAPHPNALGYHGK, has a high identity (64%) with serine/threonine kinase derived from Rhodopirellula baltica (order Planctomycetales, marine bacteria). TGAP could potently prolong thrombin time (TT), corresponding to inhibition of thrombin (FIIa) formation. Specific factor inhibitory assay showed that TGAP inhibits FVa among the major components of prothrombinase complex. In vitro assay for direct-binding affinity using surface plasmon resonance (SPR) spectrometer indicated that TGAP could be directly bound with FVa. In addition, the binding affinity of FVa to FII was decreased by addition of TGAP in dose-dependant manner ($IC_{50}$ value = 77.9 nM). These results illustrated that TGAP might interact with a heavy chain of FVa ($FVa_H$) bound to FII in prothrombin complex. The present study elucidated that non-cytotoxic T. granosa anticoagulant protein (TGAP) bound to FVa can prolong blood coagulation time by inhibiting conversion of FII to FIIa in blood coagulation cascade. In addition, TGAP did not significantly (P < 0.05) show fibrinolytic activity and cytotoxicity on venous endothelial cell line (ECV 304).

• Proceedings of the Korean Vacuum Society Conference
• /
• 2014.02a
• /
• pp.429.2-429.2
• /
• 2014

ZnO nanowire is known as synthesizable and good mechanical properties. And, stimuli-responsive polymer is widely used in the application of tunable sensing device. So, we combined these characteristics to make precise tunable sensing devise. In this work, we investigate the dependence of ZnO nanowire alignment and morphology on si substrate using nanosphere template with various conditions via hydrothermal process. Also, pH-temperature dependant tuning ability of nanostructure was studied. The brief experimental scheme is as follow. First, Zno seed layer was coated on a si wafer ($20{\times}20mm$) by spin coater. And then $1.15{\mu}m$ sized close-packed PS nanospheres were formed on a cleaned si substrate by using gas-liquid-solid interfacial self-assembly method. After that, zinc oxide nanowires were synthesized using hydrothermal method. Before the wire growth, to specify the growth site, heat treatment was performed. Finally, NIPAM(N-Isopropylacrylamide) was coated onto as-fabricated nanostructure and irradiated by UV light to form the PNIPAM network. The morphology, structures and optical properties are investigated by FE-SEM(Field Emission Scanning electron Microscopy), XRD(X-ray diffraction), OM(Optical microscopy), and WCA(water contact angle).

• Journal of Microbiology and Biotechnology
• /
• v.19 no.1
• /
• pp.17-22
• /
• 2009

The mixotrophic growth with methanol plus thiosulfate was examined in nutrient-limited mixotrophic condition for Methylobacterium goesingense CBMB5 and Methylobacterium fujisawaense CBMB37. Thiosulfate oxidation increased the growth and protein yield in mixotrophic medium that contained 150mM methanol and 20mM sodium thiosulfate, at 144 h. Respirometric study revealed that thiosulfate was the most preferable reduced inorganic sulfur source, followed by sulfite and sulfur. M. goesingense CBMB5 and M. fujisawaense CBMB37 oxidized thiosulfate directly to sulfate, and intermediate products of thiosulfate oxidation such as polythionates, sulfite, and sulfur were not detected in spent medium and they did not yield positive amplification for tested soxB primers. Enzymes of thiosulfate oxidation such as rhodanese and sulfite oxidase activities were detected in cell-free extracts of M. goesingense CBMB5, and M. fujisawaense CBMB37, and thiosulfate oxidase (tetrathionate synthase) activity was not observed. It indicated that both the organisms use the "non-S4 intermediate" sulfur oxidation pathway for thiosulfate oxidation. It is concluded from this study that M. goesingense CBMB5, and M. fujisawaense CBMB37 exhibited mixotrophic metabolism in medium containing methanol plus thiosulfate and that thiosulfate oxidation and the presence of a "Paracoccus sulfur oxidation" (PSO) pathway in methylotrophic bacteria are species dependant.

• Journal of the Korean Ceramic Society
• /
• v.36 no.7
• /
• pp.734-741
• /
• 1999

Al2O3/t-ZrO2 particulate composites were prepared by sintering at 150$0^{\circ}C$ and 1$600^{\circ}C$ for 2h in air and microstructure and mechanical properties of the composites were investigated. Although most ZrO2 particles existed at Al2O3 grain boundaries a few ZrO2 particles within Al2O3 grains. Al2O3 grain growth was depressed due to the pinning effect by ZrO2 particles. During sintering coarsening of intergranular ZrO2 particles occurred as a results of the elimination of ZrO2 intraagglomerate grain boundaries and the coalescence of dragged ZrO2 particles by migrating Al2O3 grain boundries. Changes in mechanical properties of Al2O3 composites were dependant on microstructure of Al2O3 matrix and on size and structure of dispersed ZrO2.

• Analytical Science and Technology
• /
• v.9 no.1
• /
• pp.35-42
• /
• 1996

A rapid analytical method based on capillary electrophoresis is described for the determination of trace anions in high-purity chemicals which is used to prevent corrosion demage in nuclear power plants. Separations are carried out at 20kV using trimethylsilane-coated fused-silica capillary ($70cm{\times}50$ or $75{\mu}m$ i.d.) with the electrolyte of 5mM Chromate(pH=8). Detection was achieved using on-column indirect photometry at 254nm. The simultaneous analysis of inorganic anions, chloride, nitrate, sulfate, azide and phosphate was performed using methods of hydrodynamic(>1ppm) and / or electrokinetic(<1ppm) injection. The results of studies on the coexisting anions on analyte ions shows that peak responses of analyte in hydrodynamic injection is constant without effect of coexisting anions, but those of analysis in electrokinetic injection is strongly dependant upon the kind of coexisting anions and its ionic mobility. The analyte enrichment rate, hence peak response, is positive relationship with the resistance of the sample solution. Thus, appropriate measures, such as standard addition or internal standard technique, must be used to account for differences in conductance of standard and sample solutions.

• Korean Journal of Poultry Science
• /
• v.36 no.2
• /
• pp.177-186
• /
• 2009

It is well-known that unregulated over-expression of foreign gene may have unwanted physiological or toxic effects in transgenic animals. To circumvent these problems, we constructed retrovirus vector designed to express the foreign gene under the control of the tetracycline-inducible promoter. However, gene expressions in the tetracycline-inducible expression system (Tet system) are not completely regulated but a little leaky due to the inherent defects in conventional Tet-based systems. A more tightly controllable regulatory system can be achieved when the advanced versions ($rtTA2^SM2$) of rtTA and a minimal promoter in responsive components (pTRE-tight) are used in combination therein. In this study, we tried to produce human thrombopoietin (hTPO) from various target cells and transgenic chickens using the retrovirus vector combined with Tet system. hTPO is the primary regulator of platelet production and has an important role in the survival and expansion of hematopoietic stem cells. In a preliminary experiment in vitro, higher hTPO expression and tighter expression control were observed in chicken embryonic fibroblast (CEF) cells. We also measured the biological activity of the hTPO using Mo7e cells whose proliferation is dependant on hTPO. The biological activity of the recombinant hTPO from CEF was higher than both its commercial counterpart and hTPO from other target cells. The recombinant retrovirus was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 138 injected eggs, 15 chicks hatched after 21 days of incubation. Among them, 8 hatched chicks were hTPO positive. When the Go transgenic chicken was fed doxycycline (0.5 mg per 1 gram of feed), a tetracycline derivative, hTPO concentration of the transgenic chicken blood was 200 ng/mL. Germline transmission of the transgene was confirmed in sperm of the Go transgenic roosters. These results are informative to establish transgenic chickens as bioreactors for the mass production of commercially valuable and biological active human cytokine proteins.

• Applied Biological Chemistry
• /
• v.38 no.4
• /
• pp.364-369
• /
• 1995

Anticarcinogenic activity of Moutan radix for mouse ascites cancer induced by mouse Sarcoma 180 (S-180) cells was investigated. Methanol extract of Moutan radix including other folk medicinal plants (Taxus cuspidata, Curcuma longa, Artemisia capillaris, Ligrstri fructus, and Liriope platyphylla) used to remedy or cure many chronic human diseases like cancer was fractionated into hexane, chloroform ($CHCl_3$), ethylacetate (EtOAc), and butanol (BuOH) fractions. Anticarcinogenic activity of the fractions, exhibited a strong cytotoxicity for L1210 and S-180 cells, was examined for mouse ascites cancer induced by S-180 cells. Male ICR mice (7 mice/treatment, $5{\sim}6$ weeks of age, $23{\pm}1\;g$ were injected i.p. with S-180 cells ($1{\times}10^{7}\;cell/1\;ml$ PBS). One day later, each mouse was given 0.1 ml of 10% DMSO containing sample ($30\;{\mu}g/g$ body weight) every day for 10 consecutive days. Control mice were only given 0.1ml S-180 cells and 0.1 ml 10% DMSO. Mice treated with EtOAc fraction of Moutan radix showed 28.7 days of life, which is 167% of control mice's life. Based on the dose-dependant experiment mice treated with $30\;{\mu}g$ showed longer life relative to mice treated with ootherr doses (5, 15, $60\;{\mu}g$), and mice treated with $60\;{\mu}g$ exhibited toxic symptoms. Body weight of mice treated with Moutan radix was significantly reduced relative to that of control mice (p<0.05). GC-MS analysis in conjunction with silica-gel column chromatography revealed that the EtOAc fraction contained 2-methoxylphenol, benzoic acid, 1-(4-hydroxy-3-methoxyphenyl)ethanone, 8-methyl-2,4(1H,3H)pteridinedione and 2,5-furan-dicarboxylic dimethyl ester as regards to the anticarcinogenic property of the EtOAc fraction. These results suggest that Moutan radix might be included as an anticarcinogenic medicinal plant for treatment of ascites cancer.

• Journal of fish pathology
• /
• v.18 no.1
• /
• pp.67-80
• /
• 2005

Phosphoinositide-specific phospholipase $C\delta$ $PLC\delta$) plays an important role in many cellular responses and is involved in the production of second messenger. The present study was conducted to obtain the biochemical characteristics of the expressed recombinant $PLC\delta$ in E. coli cloned from Misgurnus mizolepis and partially purified $PLC\delta$ enzymes from liver tissues of M. mizolepis (wild ML-$PLC\delta$). The ML $PLC\delta$ gene was cloned and expressed under the previous report (Kim et al., 2004), and purified the recombinant protein by successive chromatography using $Ni^{2+}$-NTA affinity column and gel iltration FPLC column. The wild ML-$PLC\delta$ protein was solublized with 2 M KCI and purified by successive chromatography on open heparin-Sephagel and analytical TSKgel heparin-5PW. Both the recombinant and wild ML-$PLC\delta$ form of protein showed a concentration-dependent PLC activity to phosphatidylinositol 4,5-bis-phosphate (PIP$_2$) or phosphatidylinositol (PI). Its activity was absolutely $Ca^{2+}$- dependant, which was similar to mammalian $PLC\delta$ isozymes. Maximal PI-hydrolytic activations of recombinant and wild ML- TEX>$PLC\delta$ was at pH 7.0 and pH 7.5, respectively. In addition, the enzymatic activities of recombinant and wild ML-$PLC\delta$ were increased in concentration-dependent manner by detergent, such as sodium deoxycholate SDC), phosphatidylethanolamine (PE) and phosphatidylcholine (PC). The activities decreased in contrast by a polyamine, such as spermine. Western blotting showed that several types of $PLC\delta$ isozymes exist in various organs. Taken together our results, it suggested that the biochemical characteristics of ML-$PLC\delta$ are similar with those of mammalian $PLC\delta1$ and ${\delta}3$ isozymes.

• Journal of Animal Environmental Science
• /
• v.10 no.1
• /
• pp.47-58
• /
• 2004

This work was carried out to investigate the effects of rice hull continuously utilized and/or replenished on the composting properties and to obtain the fundamental data between an unsupported wall and a soil supported wall during the period of composting with pig slurry in winter season. There were no the temperature holding effects in soil supported wall. New compost facility design for the temperature holding effects from a soil supported wall was required. The results were as follows; 1. Composting 1㎥ of pig slurry caused to save on 0.31㎥ of bulking agent in the unsupported wall in comparison with a soil supported wall in the rice hull single addition, and 0.45㎥ in the rice hull gradual addition. 2. The pile in the rice hull single addition had a high temperature in 4 days of composting indicating $71^{\circ}C$ and had a tendency in repeating periodically between $40^{\circ}C$ and $65^{\circ}C$ till 43 days of composting. And also the temperature of the pile was maintained between $48^{\circ}C$ and $28^{\circ}C$ after 50 days of composting. The pile of a rice hull gradual addition had the lower point of the temperature high increasingly according to adding up rice hull during the 35 days of composting. 3. The pH recorded in the rice hull single addition was higher(8.35∼10.02) compared to the rice hull gradual addition(8.6∼9.8). The pile of a rice hull single addition had a tendency in abruptly decreasing pH of the unsupported wall during the period of between 0.363$\textrm m^3$ and 0.537$\textrm m^3$ as a unit of pig slurry per rice hull. EC depending upon the way in adding rice hull was changed between 1.10 mS/$\textrm {cm}^3$ and 1.87 mS/$\textrm {cm}^3$. 4. The organic matter in an unsupported wall of the hull single addition was maintained the level of 55% during the period between 0.119㎥ and 0.363㎥ as a unit of pig slurry per rice hull while in the soil supported wall between 48 and 70. Water soluble C:N ratio was maintained between 1 and 2 in the rice hull single addition, while between 1 and 3 in the rice hull gradual addition. 5. Fertilizer constituents were detected higher level in the unsupported wall than in the soil supported wall in all treatments. This was dependant upon the input of pig slurry.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.54 no.2
• /
• pp.172-180
• /
• 2009

This study was carried out to compare the physicochemical characteristics and sensory quality of Sogokju (Korean traditional rice wine) prepared with waxy rice varieties. Among tested waxy rice varieties, highest protein contents (8.11%) was observed in cv. Sangjuchal, and highest whiteness and L-value were observed in cv. Hwaseonchal, while no significant differences in gelatinization temperature could be observed among tested varieties, and cv. Boseokchal and Sinseonchal exhibited higher breakdown viscosity compared to the others. The alcohol contents of Sogokju with waxy rice varieties ranges from 13.0 to 13.4%, and the brix degree and turbidity were within the range of 20.5 to 24.6 $^{\circ}Bx$, and 0.0344 to 0.0530, respectively. The highest L-value (6.90), b-value (l.45), pH (4.79), total acidity (0.8384%), and glucose content (10.843 g/100 ml) could be observed in Sogokju made with cv. Sangjuchal. The organic acids such as succinic acid, malic acid, citric acid and oxalic acid could be detected in Sogokju. Although no variety-dependant differences in appearance, aroma, and taste could be found in sensory evaluation, Sogokju made with cvs. Sinseonchal, Haepyeongchal, and Hwaseonchal showed higher overall quality than Sogokju made with cv. Dongjinchal, which is most widely used for Sogokju used for Sogokju production nowadays.