• The Korean Journal of Physiology
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• v.24 no.2
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• pp.363-375
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• 1990

The present investigation was performed to purify bombesin-like immunoreactivity (BBS-LI) from the skin of frogs, B. orientalis inhabiting Korea. For extraction of BBS-LI, the fresh skin of 360 g from frogs was immersed in 1,800 ml of 100% methanol and then kept at $4^{\circ}C$ for 5 days. BBS-LI was partially purified by liquid chromatography using an alkaline alumina column followed by a Sephadex G-10 column. BBS-LI was further purified by using sequential HPLC of reversed phase C18 preparation, gel permeation, SP-ion exchange and reversed phase C18 analysis. BBS-LI in fractions of each step was monitored by radioimmunoassay for which bombesin antiserum with a titer of 1 : 188,800 was raised in a guinea pig. Eventually, two different BBS-LI were successfully purified and each BBS-LI showed the following character. 1) BBS-LI was well separated into two peaks in SP-ion exchange HPLC. One (BBS-LI-K1) bound to the column while the other (BBS-LI-K2) did not. 2) BBS-LI-K1, 73.8% of total BBS-LI, was not differentiated from synthetic bombesin in reversed phase C18 analytical and gel permeation HPLC. 3) BBS-LI-K2, 26.2% of total BBS-LI, eluted later than synthetic bombesin in reversed phase C18 analytical HPLC, but it eluted with a retention time identical to that of synthetic bombesin in gel permeation HPLC. 4) The two forms of BBS-LI and synthetic bombesin identically stimulated gastrin release and pancreatic exocrine secretion including volume, protein output and amylase output in anesthetized rats. It is concluded from the above results that the skin of B. orientalis contains two different forms of BBS-LI which are very identical to bombesin immunologically and biologically. In comparison with synthetic bombesin containing 14 amino acid residues, the major form shows quite similar pattern in all HPLC used in the present study, but the minor form exhibits quite different pattern in SP-ion exchange and reversed phase C18 analytical HPLG.

• Journal of the Korean Graphic Arts Communication Society
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• v.29 no.1
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• pp.23-33
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• 2011

This experimental focus to characterize luminescence properties related to CNT (Carbon Nano Tube) element dispersedly implanted in ZnS-based phosphor thin film panel fabricated by a screen printing method. More specifically FE-SEM measurements, L-V(Luminescence vs. Voltage) and photo luminescence were carried out to determine an optimum value of CNT concentration and film thickness for the thin film structure of CNT-ZnS:(Cu, Al) by the screen printing method. We confirmed that an optimum value of CNT concentration in the ZnS:(Cu, Al) film panel is about 0.75 wt% resulting that the electric conductivity is 1.6 times higher than that of pure CNT sample and showing that the luminescence intensity is increasing until the optimum concentration. Clearly, CNT is presenting in the luminescence process providing a pathway for the creation of hot electron and a channel for the electron-hole recombination but overly inserted CNT may hinder to produce the hot electron for making an avalanching process. In case of the overly doped CNT 1.0 wt% in the ZnS-based phosphor, the luminescence intensity is decreasing although the electric conductivity is exponentially increasing. Based on these results, we realized that hot electron occurred by the external electric field or exciton arose by the external photon source are reduced dramatically over the critical value of CNT concentration because CNT element provide various isolated residues in the composites of ZnS based phosphor rather than pathway or channel for the D-A(Donnor to Acceptor) pair transition or the radiative recombination of electron-hole.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.151-157
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• 2010

Barley malt produces two different $\alpha$-amylase isozymes (AMY1 and AMY2), which share up to 80% of amino acid sequence identity with each other. However, their enzymatic properties differ remarkably. In this study, five chimeric enzymes between AMY1 and 2 were constructed by staggered extension process (StEP) technique, and their enzymatic properties were characterized. According to the results, chimeric AMY-D2, D8, and E12 showed the mixed or intermediate types of calcium-dependent activity between AMY1 and 2. Meanwhile, only AMY-E10 chimera could be significantly inhibited by barley $\alpha$-amylase/subtilisin inhibitor (BASI) protein. Chimera AMY-C6 showed the same calcium-dependency as AMY1, while AMY-E10 was closely similar to AMY2. As a result, it can be proposed that some amino acid residues in the region II, III, and IV of barley $\alpha$-amylases can play very important roles in the interaction with BASI, and those in III, V, VI, and VII may partly affect on the calcium-dependent activity.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1701-1710
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• 2017

Classical swine fever virus (CSFV) is the etiologic agent of classical swine fever, a highly contagious disease that causes significant economic losses to the swine industry. The lapinized C-strain, a widely used vaccine strain against CSFV, has low growth efficiency in cell culture, which limits the productivity in the vaccine industry. In this study, a recombinant virus derived from C-strain was constructed and subjected to continuous passaging in PK-15 cells with the goal of acquiring a high progeny virus yield. A cell-adapted virus variant, RecCpp80, had nearly 1,000-fold higher titer than its parent C-strain but lost the ability to induce fever in rabbits. Sequence analysis of cell-adapted RecC variants indicated that at least six nucleotide changes were fixed in RecCpp80. Further adaption of RecCpp80 variant in swine testicle cells led to a higher virus yield without additional mutations. Introduction of each of these residues into the wild-type RecC backbone showed that one mutation, M979R (T3310G), located in the C-terminal region of E2 might be closely related to the cell-adapted phenotype. Rabbit inoculation revealed that $RecCpp40_{+10}$ failed to induce fever in rabbits, whereas $RecCpp80_{+10}$ caused a fever response similar to the commercial C-strain vaccine. In conclusion, the C-strain can be adapted to cell culture by introducing specific mutations in its E2 protein. The mutations in RecCpp80 that led to the loss of fever response in rabbits require further investigation. Continuous passaging of the C-strain-based recombinant viruses in PK-15 cells could enhance its in vitro adaption. The non-synonymous mutations at 3310 and 3531 might play major roles in the enhanced capacity of general virus reproduction. Such findings may help design a modified C-strain for improved productivity of commercial vaccines at reduced production cost.

• Microbiology and Biotechnology Letters
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• v.48 no.2
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• pp.158-166
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• 2020

A gene coding for the xylanase was cloned from Paenibacillus woosongensis, followed by determination of its complete nucleotide sequence. This xylanase gene, designated as xyn10A, consists of 1,446 nucleotides encoding a polypeptide of 481 amino acid residues. Based on the deduced amino acid sequence, Xyn10A was identified to be a modular enzyme composed of a catalytic domain highly homologous to the glycosyl hydrolase family 10 xylanase and a putative carbohydrate-binding module (CBM) in the C-terminus. By using DEAE-sepharose and phenyl-sepharose column chromatography, Xyn10A was purified from the cellfree extract of recombinant Escherichia coli carrying a P. woosongensis xyn10A gene. The N-terminal amino acid sequence of the purified Xyn10A was identified to exactly match the sequence immediately following the signal peptide predicted by the Signal5.0 server. The purified Xyn10A was a truncated protein of 33 kDa, suggesting the deletion of CBM in the C-terminus by intracellular hydrolysis. The purified enzyme had an optimum pH and temperature of 6.0 and 55-60℃, respectively, with the kinetic parameters V_max and K_m of 298.8 U/mg and 2.47 mg/ml, respectively, for oat spelt xylan. The enzyme was more active on arabinoxylan than on oat spelt xylan and birchood xylan with low activity for p-nitrophenyl-β-xylopyranoside. Xylanase activity was significantly inhibited by 5 mM Cu²⁺, Mn²⁺, and SDS, and was noticeably enhanced by K⁺, Ni²⁺, and Ca²⁺. The enzyme could hydrolyze xylooligosaccharides larger than xylobiose. The predominant products resulting from xylooligosaccharide hydrolysis were xylobiose and xylose.

• Korean Journal of Environmental Agriculture
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• v.30 no.4
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• pp.446-452
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• 2011

BACKGROUND: To investigate the dissipation patterns of 3 pesticides, azoxystrobin, difenoconazole and iprodione, on green garlic after field treatment pesticides were treated as foliar treatment by single application at recommended and double the recommended rates. METHODS AND RESULTS: Residue samples were harvested at 0, 1, 2, 5, 7 and 10 days post-treatment for azoxystrobin and 0, 1, 2, 5, 7, 10, 15 and 21 days post-treatment for difenoconazole and iprodione. After preparation the fortified samples were extracted and analyzed by gas chromotography-electron capture detector (GC-ECD) to determine the residue levels. Recoveries ranged from 87 to 109% for azoxystrobin, difenoconazole and iprodione at two different levels. The limit of Quantification (LOQ) values were 0.002 mg/kg for azoxystrobin and difenoconazole and 0.01 mg/kg for iprodione. CONCLUSION(S): Half-lives of azoxystrobin, difenoconazole and iprodione in green garlic after treatment were 1.2, 3.8 and 3.2 days at recommended and 1.4, 3.3 and 3.2 at double the recommended rate, respectively. Residue level of azoxystrobin, difenoconazole and iprodione in green garlic were below the maximum residue limits (MRLs) at 0 day, 0 day and 5 days, respectively. Therefore, these pesticide were considered that residues was satisfied to the requirement of domestic trade related to the consumer safety.

• Analytical Science and Technology
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• v.11 no.1
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• pp.73-78
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• 1998

The reverse detection heteronuclear multiple quantum coherence, HMQC study of metcyano complex of horse myoglobin(MbCN) has provided the complete assignment of hyperfine shifted resonances of heme carbons attached with proton(s). The application of HMQC experiment to the paramagnetic low-spin MbCN gives clear $^1H$ and $^{13}C$ coherences for the paramagnetic amino acid residues as well as heme side chains, and can be extended to the low-spin paramagnetic hemoprotein derivative for the assignment of natural abundance $^{13}C$ resonances. This assignment strategy can avoid possible ambiguities that may result from the sole utilization of $^1H$ nuclear Overhauser effect for the assignment of heme $^1H$ signals resonating in the diamagnetic region. The resulting 2,4-vinyl ${\alpha}$-carbons and 7-propionate ${\beta}$-carbon follow anomalous anti-Curie behavior, and are indicative of incoplanarity with heme plane. Magnetic/electronic asymmetry of heme induced by proximal histidine(His) makes spread that the hyperfine shifted heme carbon resonances over the range of 250 ppm at $25^{\circ}C$. These heme carbon resonances would be the much more sensitive probe than those of proton resonances in analyzing the nature of heme electronic structure of myoglobin.

• The Korean Journal of Pesticide Science
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• v.7 no.3
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• pp.182-187
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• 2003

In order to elucidate a degradation characteristics of herbicide mefenacet in soil, the persistence in soils was studied under laboratory conditions for $90\sim120$ days at $28^{\circ}C$. Mefenacet residues were determined from the two soils which pre-treated by sterilization and flooding, respectively. Non-sterilized upland soil was used as a control. When 70 days elapsed from application time, $55\sim63%$ of mefenacet applied were dissipated in control soils. However, $32\sim33%$ of mefenacet applied were dissipated in the sterilized soils and $33\sim35%$ was dissipated in the flooded soils. 까 lese results indicated that the degradation of mefenacet was assumed to be due to microorganism, especially aerobic microbes. In order to elucidate the influence of water content on the persistence of mefenacet in soil, water content in soils was adjusted to 20, 50, and 80% of the water-holding capacity(Field capacity, WHC). The half-life of mefenacet in soil containing 20% and 50% of WHC were 82 and 73 days, respectively, after incubation for 90 days. However, the half-life in soil containing 80% of WHC was shortened to 61 days. These results indicated that degradation of mefenacet in soil was influenced by the activity of soil microorganism, organic matter content and water content.

• The Korean Journal of Pesticide Science
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• v.9 no.4
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• pp.330-337
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• 2005

This study was investigated to elucidate the pesticide mobility in three different soil textures(Heongseong sandy loam; Chuncheon, loam; Taeback, silty clay loam) and four different gravel contents(0, 20, 40, 60%) of Taebaek soil using soil column. Carbofuran, which ranks the highest water solubility among 7 pesticides(carbendazim, carbofuran, chlorpyrifos, cypermethrin, dimethomorph, diniconazole and endosulfan) was defected over 87% in leachate samples within all soil types from early sampling time. Amount of 5 residual pesticides excluding carbendazim and carbofuran were ordered silty clay loam > loam > sandy loam, indicating pesticide residues are related to percentage of clay contents in soils. Comparing the amount of residual pesticides in soil column(upper, middle and lower layer), 6 pesticides apart from carbofuran were found in the range of $50{\sim}92%$ on the upper layer of silty clay loam and loam. Mobility of pesticides either in soil or leachate samples is dependant on water solubility of pesticide and clay content of soil. The results obtained from four different gravel contents of Tacback soil were similar to the results of three different soil textures. Also it was found that more the gravel contents, faster the flow velocity of leachate water. These results possibly provide an idea to select proper pesticides and to reduce soil and water contamination at alpine and sloped-land.

• The Korean Journal of Pesticide Science
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• v.5 no.2
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• pp.1-12
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• 2001

A competitive indirect enzyme-linked immunosorbent assay (ci ELISA) for the organophosphorus insecticide acephate, O,S-dimethyl acetylphosphoramidothioate, was developed using a polyclonal antibody. Three different haptens mimicking the analyze and containing hexanoic acid moiety as a linker were synthesized, and then conjugated with the carrier proteins bovine serum albumin and keyhole limpet hemocyanin by the N-hydroxysuccinimide active ester method. Polyclonal antibodies raised against hapten-KLH conjugates in rabbits and the hapten-BSA conjugates as coating antigens were screened and selected for the assay in the homologous and/or heterologous ELISA system. The effects of various assay conditions, including blocking reagents, detergent content, organic solvents, pH, and preincubation of tile mixture of the polyclonal antibody and the analyze on the sensitivity were evaluated. The $IC_{50}$ value of acephate of 110 ng/mL was obtained in an optimized heterologous system using hapten-3-BSA as a coating antigen and a polyclonal antibody 8377, showing the detection range of 10-1000 ng/mL and the lowest detection limit of 4 ng/mL. The cross-reactivities of the structurally related insecticides, including methamidophos were less than 0.02%. These results indicate that the ELISA could be a convenient and alternative tool for monitoring acephate residues in agricultural products and environmental samples.