• Journal of Microbiology and Biotechnology
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• v.19 no.3
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• pp.331-337
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• 2009

Interleukin-13 (IL-13) has been proposed as a therapeutic target for bronchial asthma as it plays crucial roles in the pathogenesis of the disease. We developed an in vitro test system measuring transcriptional downregulatory activities on IL-13 as a primary screening method to select drug candidates from natural products. The promoter region of IL-13 (-2,048 to +1) was cloned into the upstream of a luciferase gene in the plasmid pGL4.14 containing the hygromycin resistance gene as a selection marker, generating pGL4.14-IL-13. The EL-4 thymoma and RBL-2H3 mast cells transiently expressing this plasmid highly produced the luciferase activities by responding to PI (PMA and ionomycin) stimulation up to 8-fold and 13-fold compared with the control, respectively, whereas cyclosporin A, a well-known antiasthmatic agent, significantly downregulated the activities. The BF1 clone of RBL-2H3 cells constitutively expressing pGL4.14-IL-13 was established by selecting surviving cells under a constant lethal dose of hygromycin treatment. The feasibility of this system was evaluated by measuring the downregulatory activities of 354 natural products on the IL-13 promoter using the BF1 clone. An extract from Morus bombycis (named TBRC 156) significantly inhibited PI-induced luciferase activities and IL-13 mRNA expression, but not the protein expression. Fisetin (named TBRC 353) inhibited not only PI-induced luciferase activities and mRNA expression, but also the IL-13 protein secretion, whereas myricetin (named TBRC 354) could not suppress the IL-13 expression at all. Our data indicated that this in vitro test system is able to discriminate the effects on IL-13 expression, and furthermore, that it might be suitable as a simple and time-saving primary screening system to select antiasthmatic agents by measuring transcriptional activities of the IL-13 promoter.

• The Korean Journal of Physiology and Pharmacology
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• v.15 no.1
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• pp.61-66
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• 2011

P2Y receptors are metabotropic G-protein-coupled receptors, which are involved in many important biologic functions in the central nervous system including retina. Subtypes of P2Y receptors in retinal tissue vary according to the species and the cell types. We examined the molecular and pharmacologic profiles of P2Y purinoceptors in retinoblastoma cell, which has not been identified yet. To achieve this goal, we used $Ca^{2+}$ imaging technique and western blot analysis in WERI-Rb-1 cell, a human retinoblastoma cell line. ATP ($10\;{\mu}M$) elicited strong but transient $[Ca^{2+}]_i$ increase in a concentration dependent manner from more than 80% of the WERI-Rb-1 cells (n=46). Orders of potency of P2Y agonists in evoking $[Ca^{2+}]_i$ transients were 2MeS-ATP>ATP>>UTP=${\alpha}{\beta}$-MeATP, which was compatible with the subclass of $P2Y_1$ receptor. The $[Ca^{2+}]_i$ transients evoked by applications of 2MeS-ATP and/or ATP were also profoundly suppressed in the presence of $P2Y_1$ selective blocker (MRS 2179; $30\;{\mu}M$). $P2Y_1$ receptor expression in WERI-Rb-1 cells was also identified by using western blot. Taken together, $P2Y_1$ receptor is mainly expressed in a retinoblastoma cell, which elicits $Ca^{2+}$ release from internal $Ca^{2+}$ storage sites via the phospholipase C-mediated pathway. $P2Y_1$ receptor activation in retinoblastoma cell could be a useful model to investigate the role of purinergic $[Ca^{2+}]_i$ signaling in neural tissue as well as to find a novel therapeutic target to this lethal cancer.

• Microbiology and Biotechnology Letters
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• v.39 no.2
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• pp.140-145
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• 2011

Porcine epidemic diarrhea virus (PEDV) infection causes acute enteritis with lethal watery diarrhea resulting in a high mortality rate in piglets. As with the other members of group 1 coronaviruses, PEDV also utilizes the host aminopeptidase N (APN) as the major cellular receptor for entry into target cells. The coronavirus spike (S) protein is known to interact with the cellular surface for viral attachment and the S1 domain of all characterized coronaviruses contains a receptor-binding domain (RBD) that mediates a specific high-affinity interaction with their respective cellular receptors. Although the RBDs of several coronaviruses have been mapped, the location of the PEDV RBD has to date not been defined. As a first step toward the identification of the region of the S protein of the PEDV that is critical for recognition with the cellular receptor, we generated a series of S1-truncated variants and examined their abilities to bind to the porcine APN (pAPN) receptor. Our data indicate that the N-terminus of the S1 domain is required for pAPN association. The results from the present study may assist in our understanding of the molecular interactions between the PEDV S protein and the pAPN receptor.

• Toxicological Research
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• v.29 no.4
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• pp.263-278
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• 2013

The silkworm extract powder contain 1-deoxynojirimycin (DNJ), a potent ${\alpha}$-glycosidase inhibitor, has therapeutic potency against diabetes mellitus. Therefore, natural products containing DNJ from mulberry leaves and silkworm are consumed as health functional food. The present study was performed to evaluate the safety of the silkworm extract powder, a health food which containing the DNJ. The repeated toxicity studies and gentic toxicity studies of the silkworm extract powder were performed to obtain the data for new functional food approval in MFDS. The safety was evaluated by a single-dose oral toxicity study and a 90 day repeated-dose oral toxicity study in Sprague-Dawley rats. The silkworm extract powder was also evaluated for its mutagenic potential in a battery of genetic toxicity test: in vitro bacterial reverse mutation assay, in vitro chromosomal aberration test, and in vivo mouse bone marrow micronucleus assay. The results of the genetic toxicology assays were negative in all of the assays. The approximate lethal dose in single oral dose toxicity study was considered to be higher than 5000 mg/kg in rats. In the 90 day study, the dose levels were wet at 0, 500, 1000, 2000 mg/kg/day, and 10 animals/sex/dose were treated with oral gavage. The parameters that were monitored were clinical signs, body weights, food and water consumptions, ophthalmic examination, urinalysis, hematology, serum biochemistry, necropsy findings, organ weights, and histopathological examination. No adverse effects were observed after the 90 day administration of the silkworm extract powder. The No-Observed-Adverse-Effect-Level (NOAEL) of silkworm extract powder in the 90 day study was 2000 mg/kg/day in both sexes, and no target organ was identified.

• Korean journal of applied entomology
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• v.56 no.4
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• pp.395-402
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• 2017

Overuse of synthetic pesticides caused negative side-effects such as environmental contamination, development of insect pests' resistance, and effects on non-target organisms. Plant origin substances without/or with low mammalian toxicity have been considered as promising alternatives to the synthetic pesticides. Fumigant toxicity of the essential oil of Iranian Yarrow, Achillea millefolium L., was investigated against a cosmopolitan stored-product insect pest: saw-toothed grain beetle (Oryzaephilus surinamensis L.). Chemical profile of this essential oil was studied by Gas Chromatography-Mass Spectrometry. Tested concentrations were significantly effective to the mortality of insect pest. A positive correlation between essential oil concentrations and pest mortality were realized. LC50 value (lethal concentration needed to 50% mortality) was achieved as $17.977(16.195{\pm}20.433){\mu}l/l$ air. The main components were 1,8-Cineole (13.17%), nerolidol (12.87%), ${\alpha}$-cubebene (12.35%), artemisia ketone (6.69%), ${\alpha}$-terpineol (5.27%), alloaromadendrene oxide (4.71%) and borneol (3.99%). Terpenic compounds including monoterpene hydrocarbons (8.19%), monoterpenoids (44.23%), sesquiterpene hydrocarbons (21.69%) and sesquiterpenoids (22.24%) were 96.35% of the total identified compounds. Results indicated that the terpene-rich A. millefolium essential oil may be considered as a safe bio-agent in the O. surinamensis management.

• Korean journal of applied entomology
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• v.48 no.2
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• pp.123-158
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• 2009

An attempt was made to stimulate future research by providing exemplary information, which would integrate published knowledge to solve specific pest problem caused by resistance. This review was directed to find a way for delaying resistance development with consideration of chemical(s) nature, of mixture, rotation, or mosaics, and of insecticide(s) compatible with the biological agents in integrated pest management (IPM). The application frequency, related to the resistance development, was influenced by insecticide activity from potentiation, residual period, and the vulnerability to resistance development of chemical, with secondary pest. Chemical affected feeding, locomotion, flight, mating, and predator avoidance. Insecticides with negative cross-resistance by the difference of target sites and mode of action would be adapted to mixture, rotation and mosaic. Mixtures for delaying resistance depend on each component killing very high percentage of the insects, considering allele dominance, cross-resistance, and immigration and fitness disadvantage. Potential disadvantages associated with mixtures include disruption of biological control, resistance in secondary pests, selecting very resistant population, and extending cross-resistance range. The rotation would use insecticides in high and low doses, or with different metabolic mechanisms. Mosaic apply insecticides to the different sectors of a grid for highly mobile insects, spray unrelated insecticides to sedentary aphids in different areas, or mix plots of insecticide-treated and untreated rows. On the evolution of pest resistance, selectivity and resistance of parasitoids and predator decreased the number of generations in which pesticide treatment is required and they could be complementary to refuges from pesticides To enhance the viability of parasitoids, the terms on the insecticides selectivity and factors affecting to the selectivity in field were examined. For establishment of resistant parasitoid, migration, survivorship, refuge, alternative pesticides were considered. To use parasitoids under the pressure of pesticides, resistant or tolerant parasitoids were tested, collected, and/or selected. A parasitoid parasitized more successfully in the susceptible host than the resistant. Factors affecting to selective toxicity of predator are mixing mineral oil, application method, insecticide contaminated prey, trait of individual insecticide, sub-lethal doses, and the developmental stage of predators. To improve the predator/prey ratio in field, application time, method, and formulation of pesticide, reducing dose rate, using mulches and weeds, multicropping and managing of surroundings are suggested. Plant resistance, predator activity, selective insect growth regulator, and alternative prey positively contributed to the increase of the ratio. Using selective insecticides or insecticide resistant predator controlled its phytophagous prey mites, kept them below an economic level, increased yield, and reduced the spray number and fruits damaged.

• The Korean Journal of Physiology
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• v.3 no.2
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• pp.9-16
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• 1969

It is well known that a number of -SH and -SS containing substances afford a certain measure of protection against radiation effects in many biological systems, and it is conceivable that inherent -SH levels in Ehrlich ascites tumour (ELD)cells may be of decisive improtance with respect to the development of cellular radiation injury. So far, little effort has been directed to elucidate the changes in levels of different -SH and -SS groups in ELD cells when the tumour-bearing whole animal was subjected to the sublethal dose of X-irradiation. The present study was designed to bring some lights in the possible changes of and relationship between various sulfhydryl levels, such as P-SH, NP-SH and NP-SS, as well as the content of protein and cell volume of ELD cells, after subjecting the ELD mice to 1,200 r of X-irradiation. The animals used in this experiment were all mixed bred mice of $20{\sim}25\;gm$ in body weight (approximately 2 months old) irrespective of sex. 12 mice in one experiment were inoculated intraperitoneally with 0.2 ml of ascites tumour cells $(2{\times}10^6\;cells)$, and on the 7th day of the tumour growth, they were X-irradiated with 1,200 r, using the conventional X-ray machine under the following conditions: 200 Kv at 15 mA, 0.5 mm Cu filter, target-skin distance: 50 cm. Radiation dose was measured with the the Philip integrating dosimeter. At 24, 36, 48 and 60 hours after the X-irradiation, the mice were killed by cervical dislocation, and the tumours were taken out. Freshly withdrawn ascites tumours were placed in ice, and immediately the cell concentration was measured with the Coulter Cell Counter (Model B), and the hematocrit of the tumour cells were also determined. Cell volume was thus calculated by the cell concentration and hematocrit value. P-SH content of ELD cells was measured potentiometrically according to the method of Calcutt & Doxey, and NP-SH and NP-SS contents were measured spectrophotometrically by the method described by Ellman. Protein content of ELD cells was determined with the Folin phenol reagent by Lowry et al. Altogether, 48 experimental mice were used, and 12 mice with the only exception of X-irradiation were used as the control. Results obtained indicate that the contents of all the cellular sulfhydryl groups as well as cell volume and protein content of the ELD cells increase significantly as time progresses after the sub-lethal X-ray dose of 1,200 r was given and that all the increase is in a lineal fashion. The regression lines of the relative values, (i. e., taking each control value as 1) of all the values obtained, and the regression lines of cell volume, protein and NP-SH are identical, whereas those of NP-SS and P-SH appear to be widely seperated. However, the difference of those two lines (NP-SS & P-SH) were found to be not significant statistically (p>0.05). Therefore, it can be concluded from the above results that all the values examined increase in a lineal fashion with no statistically significant difference among them. Also, with the radiation dose of 1,200 r, the ELD cell becomes enlarged and swollen progressively up to 60 hours post-irradiation and it becomes more than two times of the original normal size at 60 hours after the irradiation, and up to this stage, it seems apparent that the cell division has been slow due to the X-irradiation applied in this experiment. It is well understandable that the contents of NP-SH, NP-SS, P-SH and protein of the ELD cells increase in parallel with the increase of the cell volume by the X-ray does used, but it also seems interesting to note that all the cellular substances tested show no appreciable difference in the pattern of increase.