RNA interference (RNAi) has been considered as an alternative strategy to control agricultural pests whereby double-strandedRNA triggers a potent and specific inhibition of its homologous mRNA. Since small double-stranded RNAs are requiredfor various RNAi applications, there is a need for cost-effective methods for producing large quantities of high-qualitydsRNA. Bacillus thuringiensis produces much insecticidal proteins with expression of their encoding genes being drivenby sporulation-dependent promoters. To develop dsRNA mass-production platform utilizing Bt, the pHT1K-EGFP plasmidvector which has cyt1Aa sporulation-dependent promoter was constructed. The transcriptional level of target gene (EGFP)is higher 113 times than Bt reference gene (ssPE). It was applied to protect honeybee from Sacbrood virus, so targetgene was replaced to SBV-vp1. By ingestion of Bt-derived dsRNA to honeybee shows positive effect on SBV suppression.

Oyster mushrooms are one of the most famous mushrooms for foods in Korea. ‘Santari’ developed on this study is P. pulmonarius of oyster mushroom species. ‘Santari’ is a new variety of oyster mushroom for the bottle culture. It was bred by mating with monokaryons isolated from ‘GMPO20404’ and ‘Hosan’. The optimum temperature for the mycelial growth was 26～29°C on PDA medium and that for the primordia formation and the growth of fruiting body of ‘Santari’ was 22°C and 20°C on sawdust media. It took 34 days to finish spawn running, 3 days to finish primordia formation, 3 days to finish fruitbody growth in the bottle culture. In the characteristics of fruit body, pilei were round type and brownish, stipe color was white and stipe shape was long and thick. The yield per bottle was 172g/1,100ml and was 43% higher than that of control variety(Hosan). As results of the physical properties of fruit body, springness, cohesive, gumminess and brittleness of stipe tissue were 87%, 82%, 193g and 16kg, respectively. The values of them are lower than those of control except cohesive.

Oyster mushrooms are one of the most famous mushrooms for foods in Korea. ‘Santari’ developed on this study is P. pulmonarius of oyster mushroom species. ‘Santari’ is a new variety of oyster mushroom for the bottle culture. It was bred by mating with monokaryons isolated from ‘GMPO20404’ and ‘Hosan’. The optimum temperature for the mycelial growth was 26~29°C on PDA medium and that for the primordia formation and the growth of fruiting body of ‘Santari’ was 22°C and 20°C on sawdust media. It took 34 days to finish spawn running, 3 days to finish primordia formation, 3 days to finish fruitbody growth in the bottle culture. In the characteristics of fruit body, pilei were round type and brownish, stipe color was white and stipe shape was long and thick. The yield per bottle was 172g/1,100ml and was 43% higher than that of control variety (Hosan). As results of the physical properties of fruit body, springness, cohesive, gumminess and brittleness of stipe tissue were 87%, 82%, 193g and 16kg, respectively. The values of them are lower than those of control except cohesive.

RNA interference (RNAi) is an universal gene-knockdown mechanism in eukaryotic organisms including insects. RNAi has been considered as an alternative strategy to control agricultural pests whereby double-stranded RNA triggers a potent and specific inhibition of its homologous mRNA. Bacillus thuringiensis (Bt) is a spore-forming bacterium that produces a copious amount of crystal proteins δ-endotoxins under the control of sporulation-dependent promoter. In order to develop dsRNA mass-production platform utilizing recombinant Bt, the pHT1K-EGFP which expresses dsRNA against EGFP under the control of Cyt1-Aa sporulation-dependent promoter was constructed and the expression level of transgene (EGFP) was confirmed by qPCR analysis. These results suggested that Bt’s potential of becoming a new platform in dsRNA production.

고준위방사성폐기물 심층처분에서 처분안전성의 신뢰도를 향상시킬 수 있는 방안으로 그리고 처분 프로그램 개발 및 인허가를 위해 많은 나라들에서 자국에 적합한 safety case를 개발하고 있다. 본 연구에서는 방사성폐기물 처분을 위한 safety case 의 의의, 필요성, 개발과정들을 정리하고 소개하였다. 그리고 처분안전성을 safety case의 다양한 측면에서 논의하였다. 아울러 스위스, 일본, 미국, 스웨덴, 핀란드 등 해외의 safety case 개발 현황과 현재 KAERI에서 개발 중인 safety case의 개발 전략을 간략히 소개하였다. 고준위방사성폐기물 처분안전성의 신뢰도 향상을 위해 safety case 기반 하에서 어떤 노력들이 필요한지를 분석하였다. 그리고 국내 상황을 반영하여 신뢰할 수 있는 정보자료의 구축, 안전성 관련 과정들의 이해, 안전성 평가의 불확실성 저감, 이해당사자와의 의사소통, 공정성과 투명성 확보 등의 실행 방안을 제안하고 논의하였다. 본 논문에 제시된 내용들은 심층처분 safety case를 이해하고, 국내에서 개발하고 있는 고준위방사성폐기물 처분 safety case 개발을 통한 처분안전성 신뢰도 향상에 기여할 수 있을 것으로 기대한다.

In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy, if it occurs. Progesterone primes the endometrium for possible implantation and inhibits uterine contraction until birth. Aldo-keto reductases (AKRs) belong to a superfamily of NADPH-dependent reductases that act on a wide range of substrates, including simple carbohydrates, steroid hormones, and endogenous prostaglandins. 20-alpha hydroxysteroid dehydrogenase (20α-HSD; EC.1.1.1.149) enzyme belongs to the family of aldo-keto reductases. 20α-HSD predominantly converts progesterone into its biologically inactive form 20α-hydroxyprogesterone (20α -OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. In addition, the activity of 20α-HSD during the luteal phase is known to be inhibited by prolactin. We have been reporting on the molecular characterizations of placental and ovarian 20α-HSD in the bovine, pig, deer and monkey. In this study, we focused on the 20α-HSD expression in testis(6, 9, 12, 18 and 21 days after birth) of miniature pig. The protein expression and localization were detected by Western blotting and Immunohistochemical analysis. 20α-HSD protein was detected at molecular weight of 37-kDa by Western blot analysis. Also the RNA expression were detected by Reverse Transcription-PCR and quantification PCR. Additionally, We are going to analysis the function and role of 20α-HSD in the pig testis.

The glycoprotein hormone family consists of follicle-stimulating hormone (FSH; GTH1), luteinizing hormone (LH; GTH2), and thyroid-stimulating hormone (TSH), which are secreted by the pituitary gland in all mammalian species, and chorionic gonadotropin, which is secreted by placental trophoblast cells in primates and equids. These hormones consist of non-covalently associated α-, β- subunits. Within a species, the amino acid sequence of α-subunit is identical across all glycoprotein hormones and is encoded by a single gene. The αβ dimer is the active form of the hormone, and biological specificity is conferred by the β-subunit. Both of α and β subunit of eel FSH has two N-glycosylation sites (α-subunit: Asn56 and Asn79; β -subunit: Asn5 and Asn22, respectively). In the present study, we constructed deglycosylated mutants at single and double sites in each subunits of eel FSH for identification of Asn linked oligosaccharides' biological role. Mutant cDANs were cloned into pcDNA3 expression vector and transiently transfected into CHO suspension cells. The quantity of rec-eelFSHs were quantified by sandwich ELISA system, using monoclonal antibodies produced in our lab. The wild type rec-FSH protein was detected at the predicted molecular weight of 34 kDa by western blot. The molecular weight of deglycosylated mutants at single site decreased with about 4 kDa and of mutants at double sites decreased with 8 kDa. After PNGase treatment in the rec-eel FSH proteins, molecular weight also decreased to 7-8 kDa. We generated stably parental cell lines, engineered to express a β-arrestin 2EA fusion protein, expressing eel FSHR and C-terminal deleted mutant. 2 out of 5 receptor cells each were selected by G-418 and we tested these cell lines in a receptor functionality using PathHunter β arrestin assay (DiscoverX). Follicle stimulating hormone acts through binding to its specific receptor. Binding of ligand to the receptor activates the adenosine 3',5'-cyclic monophosphate (cAMP) pathway (McFarland et al., 1989; Ji and Ji, 1991a; Rose, 1998) and the inositol 1phosphate (IP1) the second messenger systems. After stimulation of eelFSH receptor stably transfected Parental CHO cells with FSH wild type and mutant hormones as a ligand, production of cAMP and IP-1 were evaluated (Cisbio). cAMP IC-50 values by eelFSHwt; αΔ56; αΔ79; αΔ56_79; and βΔ5 were 33.1; 1154.7; 22; 410 and 311.9 ng/ml, respectively. IP-1 IC-50 values by eelFSHwt; αΔ56; αΔ79; αΔ56_79 and βΔ5 were 6.8; 7.1; 4.4; 3.8 and 10.2 ng/ml, respectively too. The cAMP activation was greatly decreased in the αΔ56αmutant. Thus, the site of α56 oligosaccharide in the eel plays an pivotal role for the cAMP stimulation using eel FSH receptor cell lines. In the IP-one assay, the activity in the αΔ56 and βΔ5 mutants was a little decreased than the wt. The biological roles of N-linked oligosaccharides in GPCR internalization are going to be estimated by measuring β arrestin recruitment system.

The glycoprotein hormone family consists of luteinizing hormone (LH), follicle stimulating hormone (FSH) and thyroid stimulating hormone, which are secreted by the pituitary gland in all mammalian species, and choriogonadotropin (CG), which is secreted by the placenta in primates and equids. The hormones are composed of a common α subunit and a hormone specific β subunit which are non-covalently associated. Recent advances in biotechnology, particularly in the production of recombinant proteins, have provided opportunities to produce sufficient quantities of recombinant fish GTHs using various expression hosts. Japanese eel Anguilla japonica is one of the most important fish species being aquacultured in Japan but is hampered from the fact that this species does not reproduce in captivity. Artificial induction of gonadal maturation has been successful by administration of pituitary extracts or human chorionic gonadotropin, but the understanding the regulatory mechanism of gonadal development moderated by follicle stimulating hormone (FSH) and luteinizing hormone (LH) remains elusive due to lack of suitable amounts of eel gonadotropins (GTHs). In the present study, we produced tethered rec-eel LH and deglycosylated mutants (56, 79 and 56-79 of α subunit; 10 of β-subunit) of Asn-linked oligosaccharides in CHO suspension cells. Luteinizing hormone acts through binding to its specific receptor. Binding of ligand to the receptor activates the adenosine 3',5'-cyclic monophosphate (cAMP) pathway (McFarland et al., 1989; Ji and Ji, 1991a; Rose, 1998) and the inositol 1 phosphate (IP1) secondary messenger systems. After stimulation of eelLH/CG receptor transfected CHO cells with rec-LH wild type (wt) and mutant hormones as a ligand, production of cAMP and IP-1 were evaluated (Cisbio). cAMP IC-50 values by rec-eelLH wt; αΔ56; αΔ79; αΔ56_79 and βΔ10 were 606.2; 374.9; 100.3; 14.2 and 210.9 ng/ml, respectively. IP-1 IC-50 values by rec-eelLH wt; αΔ56; αΔ79; αΔ56.79 and βΔ10 were 28.3; 16.04; 4.3; 2.1 and 3.6ng/ml, respectively too. As seen in both of the second messenger production, general stimulatory pattern is analogous. cAMP and IP-1 stimulation by wild type and αΔ56, as well as αΔ79 and βΔ10 were approximate, but the stimulating effect of double mutant (αΔ56_ 79) was drastically higher. According to the data, deglycosylated eelLH may bind to the receptor with high affinity and cAMP production is gradually increased. Furthermore, receptor activation by tethered rec-eel mutant ligands (FreeStyle CHO-MAX Expression System) will be evaluated with β arrestin recruitment and GPCR internalization for N-linked oligosaccharides’ biological role in activation of eelLH/CGR.

Streptomyces is the largest genus of Actinobacteria that forms fungus-like branched networks of hyphae. Streptomyces has been clinically important because they produce various secondary metabolites with antibacterial, antifungal, and nematocidal activities. In order to explore novel insecticidal compounds, extracts from 363 strains of Actinobacteria were screened for their juvenoid and anti-juvenoid activities using yeast-two hybrid system. Among them, extract of Streptomyces spp. showed high anti-juvenoid activity. This extract also showed high level of insecticidal activities against larvae of Aedes albopictus, Laodelphax striatellus, and Ostrinia furnacalis. These results suggested that the secondary metabolites of Streptomyces could be natural sources of novel insecticidal compounds.

4(5)-methylimidazole (4(5)-MI) is carcinogenic, nitrogen-containing compound, mainly found during the manufacturing of caramel coloring. Hence, presence of 4(5)-MI is well-known in any food products with addition of caramel coloring for desirable sensory characteristics, such as cookies. Limited work has been conducted to develop suitable analytical method and to investigate effect of caramel coloring in cookies. This study aimed at developing the analytical method for quantification of 4(5)-MI and confirming influence of caramel coloring on level of 4(5)-Mi in cookies. Gas chromatography-mass spectrometry (GC-MS) was utilized for qualification and quantification of 4(5)-MI. Sample preparation procedure specialized in bakery products was fully developed in this study. The concentration of 4(5)-MI in 15 commercial cookies and biscuits ranged from 71.5 to 1254.8 ng/g. Correlation equation (y = 706.42x + 21.792) was obtained to estimate effect of caramel colorant on level of 4(5)-MI in cookies. Further, analytical method developed and results of correlation equation can be utilized in future studies on reduction of 4(5)-MI in many food.

This study performed to investigate the changes in S-allyl-L-cysteine (SAC) content of garlic with different cooking methods. Methods for determining SAC was validated by determining specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision and accuracy using HPLC-FLD system. Results showed high linearity in the calibration curve with a coefficient of correaltion (R 2 ) of 0.9999. The LOD and LOQ values for SAC were 0.15 and 0.49 μg/mL, respectively. The relative standard deviations (RSDs) for intra- and inter-day precision of SAC were less than 5%. The recovery rate of SAC was in the range from 97.35% to 97.47%. The SAC content of raw garlic was 2.77 mg/g, and there are no significantly difference among blanching and microwave treatment in SAC content. However, SAC content of boiling, autoclaving at 110℃ and 121℃ increased in the range of 3.50~9.19 mg/g, 6.52~16.21 mg/g and 14.15~50.24 mg/g as increasing cooking temperature and time, respectively.

The present study was made to assess annual changes on seasonal occurrence of Apiformes, commonly known as bees, as global warming apparently happens. Bees were collected from ten sites using a malaise trap for each site, and sample collection was made every two weeks from April to September in 2011 to 2014. In overall diversity, abundance and richness of bees appeared higher in northern sites, particularly Gwangneung and Hongcheon, than southern. The abundance of bees declined sharply in 2013 and decreased even further in 2014. Among the bee families the most decreased families were Halictidae and Andrenidae. In order to see the effect of annual temperature fluctuation correlation analysis was used with collection data and weather data. However, the result indicated that the decline of the bees was not significantly affected by changes of temperature although most of the bee families except Halictidae showed negative relation with temperature data(p < 0.05). In addition we also analysed the bee collection data and flowering data during spring in Gwangneung. Dufrene-Legendre indicator species analysis (ISA) for latitudinal sites identified seventeen indicator species (p < 0.05) that were limited to northern region.

This review presents current progress in the preparation methods of liquid crystalline nanocarbon materials and the liquid crystalline spinning method for producing nano-carbon fibers. In particular, we focus on the fabrication of liquid crystalline carbon nanotubes by spinning from superacids, and the continuous production of macroscopic fiber from liquid crystalline graphene oxide.

Bean bug, Riptortus pedestris is an agriculturally serious pest in north eastern Asian countries, damaging to several legumes and fruit trees. Chemical pesticides have been largely used to control the pest but it encounters insecticide-resistance and environmental toxicity issues. Alternatively different mode of action and environmentally sound pest management system can be found in entomopathogenic fungal insecticides. Herein we developed a platform to optimize the fungal production to express their maximum virulence against bean bug, by focusing on solid culture system for thermotolerance, formulation to select effective surfactants to carry the fungal conidia on the cuticles, and relationship between environmental abiotic factors and fungal mortality. First to produce highly thermotolerance fungal conidia, Beauveria bassiana and Metarhizium anisopliae isolates were cultured on several granular cereal substrates, which could be subjected to formulation process. Among the tested media, four media (millet, non-glutinous italian foxtail millet, glutinous italian foxtail millet, brown rice) were superior to the other grains in the spore production and thermotolerance. Next to efficiently deliver the fungal conidia on the cuticles of bean bug, total of six surfactant (CO-2.5, CO-12, LE-7, PE-61, TED-3 and Siloxane) was used to experiment. CO-12 was superior to the other surfactant in mortality of 100 ppm consistence. This work suggests that solid culture system and formulation and application should be seriously considered to reach an optimal level of mortality by inducing their maximum virulence.