• Applied Biological Chemistry
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• v.46 no.4
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• pp.353-360
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• 2003

For the simple rapid bioassay of organophosphorus and carbamate pesticide residues, a mass-production system of acetycholinesterase (AChE, EC 3.1.1.7, MAChE) using baculovirus and insect cell culture was constructed. The cDNA for AChE was synthesized from Drosophila melanogaster in Halla Mountain, the lipid anchor tail was removed by PCR and was used for the site-directed mutagenesis of three amino acid residues (E107Y, F368L, L408F). The mutated cDNA was inserted into the baculovirus vector and expressed in insect cells. Maximum cell growth and enzyme activity were reached when the cells $(2{\times}10^6\;cell/ml)$ were infected four times at four-day-intervals. His-tag containing MAChE was purified using Ni-NTA column and used for characterization. The activity was maintained under various pHs (3-10) and temperatures $(20-50^{\circ}C)$ under experimental conditions. As an extraction solution for pesticides, methanol is more effective than ethanol. Against major organophosphate and carbamate pesticides, the MAChE showed better sensitivity than AChE and AChE from housefly (Taiwan).

• Korean journal of applied entomology
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• v.35 no.3
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• pp.232-237
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• 1996

Toxicity of 5 acaricides, 4 insecticides. and 6 fungicides to the twospotted spider mite, Tetranychus urticae Koch, and its predator Amblyseius womersleyi Schicha was assessed in a laboratory using a leaf disk bioassay. Dicofol was equally very toxic to aduly females of T. urticae and A. womersleyi, and caused 46% and 40% mortality of eggs of those two species, respectively. Adult females of A. womersleyi dipped in Abamectin solution showed low mortality(16.6%), while all T. urticae females died within 24 hours after dipping. Three ovicidal acaricides (Tetradifon, Clofentezine, and Hexythiazox) showed no mortality of predator eggs, but more than 90% mortality of T. urticae eggs. Four insecticides (Teflubenzuron, Triflumuron, Diflubenzuron, and Imidacloprid) and three fungicides (Propineb, Fenarimol, and Polyoxin-B captan) were non toxic to the A. womersleyi adult females. The other fungicides (Chlorothalonil, Nuarimol.Mancozeb, and Folpet) were a little toxic to adult females of the predator, showing$\leq$23.3% mortality, It may be suggested from these results that 4 insecticides, 4 acaricides, and 6 fungicides described could be incorporated into the integrated pest management system with A. womersleyi in apple orchard.

• Environmental Mutagens and Carcinogens
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• v.24 no.4
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• pp.198-205
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• 2004

Cytochrome P4501B1(CYP1B1) is known to be inducible by xenobiotic compounds such as policyclic aromatic hydrocarbon(PAH) and dioxins such as 2,3,7,8-tetrachloro-dibenzo-p-dioxin(TCDD). And these induction of CYP1B1 is also regulated by many categories of chemicals. In order to investigate the effects of several chemicals on CYP1B1 gene expression in Hepa-I and MCF-7 cells, 5' flanking DNA of human CYP1B1 was cloned into pGL3 basic vector containing luciferase gene, and then transfected into these cells. After treatment of chemicals, the luciferase activity was measured. CYP1B1 enzyme metabolize PAHs and estradiol. CYP1B1 metabolize estradiol to 4-hydrozyestradiol that is considered as carcinogenic metabolite. Recent industrialized industrialized society, human has been widely been exposed to widespread environmental contaminants such as PAHs(polycyclic aromatic hydrocarbon) that are originated from the imcomplete combustion of hydrocarbons. PAHs are known to be ligands of the AhR(aryl hydrocarbon receptor). Induction of cytochrome P4501B1(CYP1B1) in cell culture is widely used as a biomarker for PAHs. Therefore we have studied the effect of PAHs in the human breast cancer cells MCF-7 to evaluate bioactivity of PAHs. We have used the United State of America EPA selected 13 different PAHs, PAHs mixtures and extracts from environmental samples to evaluate the bioassay system. We examined effects of PAHs on the CYP1B1-luciferase reporter gene and CYP1B1 mRNA level. Benzo(k)fluoranthene and dibenzo(a, h)anthracene showed strong response to CYP1B1 promoter activity stimulation, and also CYP1B1 mRNAs increase in MCF-7 cells in a concentration-dependent manner. RT-PCR analysis indicated that PAHs significantly up-regulate the level of CYP1B1 mRNA. Some flavonoids such as genistein, daidzein, chrysin, naringenin and morin were also investigeted. These flavonoids decreased B(k)F infuced luciferase activity at low concentration. But, these flavonoids exhibited stimulatory effect at high concentration.

• KSBB Journal
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• v.24 no.2
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• pp.163-169
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• 2009

Lovastatin (also known as Mevinolin, Mevacor, and Monacolin K), an inhibitor of the HMG-CoA reductase produced by Aspergillus terreus and other fungi, is used to reduce serum cholesterol levels in human beings. It is derived biosynthetically from two polyketides. One of these is a nonaketide that undergoes cyclization at a hexahydronaphthalene ring system, and the other is a simple diketide, 2-methylbutyrate. Two primer pairs were designed based on the amino acid sequences of lovastatin polyketide synthase and lovastatin diketide synthase for the PCR screening of lovastatin-producing strains. Among the seven selected strains, SJ-2 evidenced the highest level of lovastatin production in both liquid and solid cultures. Soybeans with SJ-2 were treated via 1 hour of heat shock at $30^{\circ}C$ for the mass production of lovastatin. The heat-treated soybeans were inoculated on rice bran and the koji extract was obtained after 15 days of incubation. It yielded the highest level of lovastatin production among the strains, and also evidenced 75% inhibition activity against HMG-CoA reductase. We developed an efficient PCR screening method for lovastatin-producing strains, using lovastatin biosynthesis genes.

• Food Science and Preservation
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• v.23 no.7
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• pp.989-994
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• 2016

As part of our continuing search for bioactive natural products, the antioxidant and ${\alpha}$-glucosidase inhibitory activities of an 80% methanolic extract and organic solvent soluble-portions of Aruncus dioicus var. kamtschaticus roots were investigated by using a bioassay system. The antioxidant activity of A. dioicus var. kamtschaticus roots extract and organic solvent soluble-portions were assessed by examining with 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) ($ABTS^+$) radical scavenging. In addition, anti-diabetic effects of the A. dioicus var. kamtschaticus root extract and organic solvent soluble-portions were tested via ${\alpha}$-glucosidase inhibition assay. The total phenolic contents of the products were determined by applying UV-VIS spectrophotometry. All tested samples showed dose-dependent radical scavenging and ${\alpha}$-glucosidase inhibitory properties. In particular, the ${\alpha}$-glucosidase inhibitory and radical scavenging effects of the ethyl-acetate (EtOAc)-soluble portion from the roots of A. dioicus var. kamtschaticus were greater than those from other solvent-soluble portions. These results indicate that A. dioicus var. kamtschaticus could be considered a new effective source of natural antioxidants and anti-diabetic materials. More systematic research of the constituents of the roots of this A. dioicus variety will be conducted to further develop its antioxidative and anti-diabetic properties.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.64-64
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• 2019

Screening and identification of genetic resources based on their phytoconstituents and morphological characters potentially provide baseline data for researchers, breeders, and nutraceutical companies who wish to formulate a nutrient-dense diet and health beneficial supplement. Thus, we evaluated the amount of total phenolic content and major phenolic compounds; examined if phenolic compounds could be used as distinguishing factors for perilla genetic resources; and investigated the relation between some quantitative and qualitative morphological characters with the contents of phenolic compounds in 360 accessions obtained from National Agrobiodiversity Center gene bank, Jeonju, Korea. Total phenolic content (TPC) was estimated using Folin-Ciocalteu colorimetric assay. Individual phenolic compounds were determined using an Ultra-High Performance Liquid Chromatography system equipped with Photodiode Array detector. Considerable variations were observed in TPC (7.99 to 117.47 mg GAE/g DE), rosmarinic acid (RA) (ND to 19.19 mg/g DE), caffeic acid (CA) (ND to 0.72 mg/g DE), apigenin-7-O-diglucuronide (ADG) (ND to 1.24 mg luteolin equivalent (LUE)/g DE), scutellarein-7-O-glucuronide (SG) (ND to 4.32 mg LUE/g DE), and apigenin-7-O-glucuronide (AG) (ND to 1.60 mg LUE/g DE). RA was the most dominant phenolic compound in most accessions (95.3%) followed by SG. The adaxial leaf color was light green, green and dark green in 13.8%, 65.0%, and 21.1 % of the accessions, respectively. 78.8% of the accessions had light green color at the abaxial side with the remaining being described as green. Most of the accessions (96.9%) were cordate shape, the remaining being eclipse. Intensities of green pigment at abaxial and adaxial leaf surfaces were correlated with contents of individual phenolic compounds and TPC whereas leaf length and width had no correlation with TPC, CA and RA, and negatively correlated with ADG, AG, and SG. Leaf shape was not related with content of phenolic compounds, color of leaves, or the length or width of leaves. Accessions IT57426, IT157434, IT267710, and IT267712 which contained relatively high contents of TPC and major phenolic compounds (RA and SG) could be used for further research in breeding and bioassay test. Our study result showed the contents of total phenolics and individual phenolic compounds along with the morphological characters could be useful distinguishing factors for perilla genetic resources.

• Journal of Practical Agriculture & Fisheries Research
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• v.22 no.2
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• pp.135-146
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• 2020

In the strawberry cultivation areas and domestic farms, two-spotted spider mites are the most serious pests. It decreases the product yield due to the direct feeding of chlorophyll by the mites resulting in reduced photosynthetic ability of host plant. In this study, a simplified acaricidal effect evaluation system (SAEES) was employed to choose the effective acaricidal products among 10 items based on leaf-dipping bioassay methods. SAEES had the advantage of being able to screen four commercial products with three replications at the same time in the recommended concentration. The susceptible strains (SL_YS) showed a high mortality rate of about 90-100%, whereas the mortality of local strains was differed by each acaricides. It suggests that the acaricide responses of field populations might differ due to spray frequencies and acaricide product. An efficacy index (0.8-1.0) was determined based on the mortality of susceptible strain, which would allow the most effective commercial products to be selected by the range of this index. In summary, SAEES will enable the selection of effective commercial products and contribute to increasing control against T. urticae in strawberries.

• Journal of Sericultural and Entomological Science
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• v.23 no.1
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• pp.1-31
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• 1981

Rootability of the hardwood cuttings of mulberry was related not only histological characteristics but dependent on biochemical properties. In this connection, the characteristics of the hardwood cuttings were histologically observed and the growth substances produced by the cuttings were also identified by means of mung bean bioassay. Amino acid, carbohydrate, nucleic acid contents, and the C/N ratio were also analysed. The results are summarized as follows. 1. There were differences in rootability of cuttings between mulberry species and varieties Among the three mulberry species tested, Morus Lhou Koidz. showed the highest rootability while M. bombycis showed the lowest one. In varietal differences in rootability, it was shown that the varieties could be grouped according to rootability: high varieties(above 80%), medium(41~79%), and low(below 40%). The higher varieties were Kemmochi, Nakamaki, Kosen, and Wusuba roso. 2. The histological characteristic of the hardwood cuttings most closely related to rootability was cell layer arrangement in the sclerenchyma tissue. The lower rootability varieties developed two or three overlapping cell layers in the bark tissue and in the higher rootability varieties they were scattered over the primary cortex. 3. In the higher rootability varieties, there was a positive correlation between the development of root primodia and rootability of the hardwood cuttings. It was also shown that there was a close relationship between the size of primodia and the surface area of the lenticel with rootability of the cuttings. 4. Effect of growth substances extracted from the hardwood cuttings were determined by mung bean bioassay. The higher rootability varieties usually showed higher activities of the growth substances, in contrast the lower rootability varieties showed higher activities of the inhibitory substances. 5. It was evident that the substance separated by paper chromatography was identified as indole acetic acid with $R_f$ value ranging from 0.3 to 0.5. The other substances detected at a $R_f$ value ranging from 0.8 to 1.0 and origin to 0.1 were also responsible for rooting. 6. There exists a quantitatively different distribution of growth substances in a synergistic system in the tissues of cuttings, and the balance between growth and inhibitory substances gives rise to the development of rooting. Particularly, no descent of the substances from winter buds resulted in no rooting of cuttings but these substances were produced a week after planting in a warm environment. 7. It was shown that there were positive correlations between carbohydrate ($r=0.72^*$) and total sugar ($r=0.67^*$) and rootability, respectively, but there were negative correlations between reducing sugars ($r=-0.75^*$) and rootability. 8. High C/N ratio gave rise to high rootability($r=0.67^*$). The latter therefore depended on high amount of carbohydrate rather than nitrogen in the cuttings. 9. The content of RNA and DNA in the cuttings was not changed for upto two weeks after the cuttings were planted. Then an increase in RNA content took place in only the high rootability varieties. 10. There were quantitative and qualitative differences in the compositions of the amino acids between the high rootability varieties and the low rootability varieties. More aspartic acid and cystine were found in the higher rootability varieties than in the low rootability varieties.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.299-308
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• 2002

In horse, a single gene encodes both eCG and eLH $\beta$ subunits. The difference between eCG and eLH lies in the structure of their glycoresidues, which are both sialylated and sulfated in LH and sialylated in CG eCG consists of highly glycosyiated $\alpha$- and $\beta$-subunits and is an unique member of the gonadotropin family because it elicits response characteristics of both FSH and LH in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of gonadotropin structure-function relationships and the understanding of the molecular bases of the specific interactions of these hormones with their receptors. Thus, eCG is a dintinct molecule from the view points of its biological function and glycoresidue structures. The oligosaccharide at Asn 56 of the $\alpha$-subunit plays an indispensable role, whereas the carboxyl-terminal extension of the eCG $\beta$-subunit with its associated O-linked oligosaccharides is not improtant for, the in vitro LH-like activity of eCG. In contrast, both N- and O-linked oligosaccharides play important roles for FSH-like activity and increase FSH-like activity by removal of N- and O-linked oligosaccharides. Therefore, the dual LH- and FSH-like activities of eCG can be clearly separated by removal of either the N-linked oligosaccharide on the $\alpha$-subunit or CTP-associated O-linked oligosaccharides from its $\beta$-subunit. The glycoresidues seem to play crucial roles fer biological activities. The tethered-eCG was effciently secreted and showed similar LH-like activity to the dimeric eCG $\alpha$/ $\beta$ and native eCG. FSH-like activity of the tethered-eCG was also shown similarly in comparison with the native and wild type eCG $\alpha$/ $\beta$. Our data for the first time suggest that the tethered-eCG can be expressed efficiently and the produced product by the CHO-Kl cells is fully LH- and FSH-like activities in rat in vitro bioassay system. Our results also suggest that this molecular can imply particular models ot FSH-like activity not LH-like activity in the eCG. Taken together, these data indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion.

• The Korean Journal of Pesticide Science
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• v.4 no.4
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• pp.40-47
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• 2000

The comparative toxicity of ten acaricides, seven insecticides and five fungicides to the two-spotted spider mite, Tetranychus urticae and its predators, Amblyseius womersleyi and A. eharai was evaluated by a leaf spray bioassay. Five of the acaricides tested, bifenazate, etoxazole, acequinocyl, flufenoxuron and chlorfenapyr were much less toxic to adult females of A. womersleyi and A. eharai than to T. urticae adult females. A. womersleyi adult females treated with five acaricides produced $52{\sim}93%$ as many eggs as untreated adult females. And A. eharai adult females treated with five acaricides laid $54{\sim}73%$ as many eggs as untreated adult females. The remaining acaricides showed high toxicity to adult females of A. womersleyi and A. eharai. All the insecticides tested were less toxic to T. urticae adult females than to adult females of A. womersleyi and A. eharai. However, tebufenozide and diflubenzuron did not significantly affect the survival and reproduction of adult females of A. womersleyi and A. eharai. All the fungicides tested showed low mortality (${\leq}24%$) to adult females of predatory mites. However, benomyl had significant effect on the reproduction of adult females of A. womersleyi and A. eharai. Four acaricides (bifenazate, acequinocyl, flufelloxuron and chlorfenapyr) were much less toxic to eggs of A. womersleyi and A. eharai than to T. urticae eggs. However, etoxazole caused relatively low hatchability ($58{\sim}62%$) of eggs of A. womersleyi and A. eharai. All the insecticides and fungicides tested did not significantly affect the hatch of eggs of predatory mites. It may be suggested from these results that four acaricides, two insecticides and four fungicides described could be Incorporated into the integrated mite management system with A. womersleyi and A. eharai in pear orchard.