The α-Gal epitope (Galα1,3Galα1,4GlcNAc-R) is responsible for hyperacute rejection (HAR) during transgenic pig-to-non-human primate xenotransplantation. To overcome HAR after xenografts, it is essential for the inactivation of α1,3Galactosyltransferase (GT) gene by the homozygotic knocked out of GT-/- and the isoglobotrihexosylceramide synthase (iGb3s-/-). This study was performed to investigate the generation and characterization of the α1,3GT-MCP/-MCP+iGb3-/- transgenic cells. Ear fibroblast cells from the GT-MCP/-MCP pig were cultured and used to positive control. For iGb3s knock out, the Cas9-GFP-iGb3s vector was transfected into the GT-MCP/-MCP cells. The Cas9-GFP-iGb3s transfected cells were sorted and sequenced for the selection of GT-MCP/-MCP+ iGb3s-/- cells. Among the three sorted cell lines, one transgenic cell line was homozygously deleted 3 bases and 10 bases in each chromosome, respectively. To characterize an expression of α-Gal epitope, a wild type and the transgenic cells were measured by FACS Aria using BS-IB4 lectin antibody. The expression of α-Gal epitope in GT-MCP/-MCP cells (<0.01 %) were significantly down-regulated to the range of wild type (99.4 %) fibroblast cells (p<0.05). To analyze the function of iGb3s, α -Gal epitope expressions were measured for the GT-MCP/-MCP, GT-MCP/-MCP+iGb3s-/+, and GT-MCP/-MCP+iGb3s-/-. The range was 95.8%, 94.2%, and 75.8%, respectively. Interestingly, there was a negative range (16.2%) of α-Gal epitope -/- section in GT-MCP/-MCP+iGb3s-/-, compared to 2.74% of GT-MCP/-MCP+iGb3s-/+ and 1.4% of WT, respectively. Our results demonstrated that iGb3s-/-combined with GT-/- had a function to inhibit α-Gal epitope expression in pig cells. Further studies are needed to evaluate the functions of double gene knock out to minimize a HAR response after xenotransplantation.

The α-Gal epitope (Galα1,3Galα1,4GlcNAc-R) is responsible for hyperacute rejection (HAR) during transgenic pig-to-non-human primate xenotransplantation. There are genes related to the expression of α-Gal epitope such as α1,3Galactosyltransferase gene (GT-/-) and the isoglobotrihexosylceramide synthase (iGb3s-/-). This study was performed to investigate the expression of α-Gal epitope in the skin derived from GT-/- transgenic pig. The skin (7/1000 inches) was obtained by dermatome (Zimmer® Electric Dermatome) from one month old of wildtype (WT) and GT-/- piglets, respectively. The skins were fixed, dehydrated, cleaned, and embedded. To analyze the expression of α-Gal epitope, the paraffin section of WT and GT-/- were stained with BS-IB4 lectin and isoglobotrihexosylceramide synthase antibody. There was a strong BS-IB4 lectin signal in the skin of WT, but not detected in GT-/-. However, the iGb3s positive signals were stained in the skin of both WT and GT-/-. Taken together, it can be postulated that the knocked out of GT gene may not enough to inhibit the expression of α-Gal epitope. Further studies are needed to evaluate the functions of the double knock out of GT and iGb3s on the expression of α-Gal epitope.

Insect growth regulators (IGRs) are attractive pest control agents due to their high target specificity and relative safety to the environment. Recently, plants have been shown to synthesize IGRs that affect the insect juvenile hormone (JH) as a part of their defense mechanisms. We identified several JH agonists (JHAs) and antagonists (JHANs) from plant essential oil compounds using a yeast two-hybrid system transformed with the Aedes aegypti JH receptor as a reporter system. They showed high mosquitocidal activities with relatively low LC50 values and caused retardation of ovarian development in female mosquitoes. While the JHAs increased the expression of JH-induced gene, the JHANs caused reduction in the expression of the same gene. The compounds identified in this study could provide insights on the plant-insect interactions and may be useful for the development of novel IGR insecticides.

Bacillus thuringiensis (Bt) produces a variety of insecticidal crystal proteins and widely used as one of the most successful biological control agents. Recently, studies that introduce cry genes into crops to create pest resistance have made much progress, and the total area of land planted with Bt crops has increased substantially. In this study, pest resistance of 8 transgenic Bt rice events with a synthetic cry1Ac gene linked to rice rbcS-tp sequence were assessed under laboratory conditions. Bioassays were performed against Cnaphalocrocis medinalis, which is a significant pest of rice in Asia. C. medinalis larvae were shown to be susceptible to all eight events, even though there were differences between the causes of death. The results differed between developmental stages of the larvae, despite the fact that all 8 events led to high mortalities. These results may be a significant foundation for the evaluation of improved transgenic Bt rice.

Myeloid differentiation factor 88 (MyD88) is an intracellular adaptor protein involved in Toll pathway. In this study, we monitored the response of 4 key genes of the insect immune system against Beauveria bassiana JEF-007 in Tenebrio molitor using RT-PCR. To better understand the roles of Toll pathway in mealworm immune system, TmGPR and TmMyD88 was knocked down by RNAi silencing. Target gene expressions were decreased at 6 days post-dsRNA injection. Therefore, mealworms were compromised by B. bassiana JEF-007 at 6 days post-dsRNA injection. Silencing of the TmMyD88 and TmGPR resulted in reducing the resistance of the host to fungal infection. However, only dsTmMyD88 showed significant difference with dsEGFP by statistical analysis, which may be due to partial gene knock down of dsGPR. These results indicate that TmMyD88 is required in mealworms for survival against B. bassiana JEF-007.

A viral genome was assembled de novo from next-generation sequencing (NGS) data from bean bugs, Riptortus pedestris, infected with an entomopathogenic fungus, Beauveria bassiana (Bb), and was further confirmed via the RACE method. Based on the phylogenetic analysis of the RdRp sequences, RiPV-1 was clustered in the unassigned insect RNA viruses with two other viruses, APV and KFV. These three viruses were suggested to constitute a new group of insect RNA viruses. Interestingly, RiPV-1 replication was increased dramatically in bean bugs 2 to 6 days after fungal infection. In conclusion, a novel insect RNA virus was found by NGS data assembly. This virus can provide further insight into the interaction between virus, fungus and the host.

The development of nanostructured functional materials derived from biomass and/or waste is of growing importance for creating sustainable energy-storage systems. In this study, nanoporous carbonaceous materials containing numerous heteroatoms were fabricated from waste coffee grounds using a top-down process via simple heating with KOH. The nanoporous carbon nanosheets exhibited notable material properties such as high specific surface area (1960.1 m2 g–1), numerous redox-active heteroatoms (16.1 at% oxygen, 2.7 at% nitrogen, and 1.6 at% sulfur), and high aspect ratios (>100). These unique properties led to good electrochemical performance as supercapacitor electrodes. A specific capacitance of ~438.5 F g–1 was achieved at a scan rate of 2 mV s–1, and a capacitance of 176 F g–1 was maintained at a fast scan rate of 100 mV s–1. Furthermore, cyclic stability was achieved for over 2000 cycles.

Composite resins are developed as restorative materials to improve esthetics and mechanical properties. To improve the physical properties of resin material, resin filler have to be added. However, no imaging method is adopted for resin filler distribution. Optical coherence tomography (OCT) is a optical imaging technique to delineate microscopic structures within biological tissue. The OCT application to dental composites resin and its filler is not described yet. So, this new and advanced optical method is needed for clinical application for evaluation of dental composite resin. To analyze the spatial distribution of dental composite resin and to evaluate the resin restoration in cavity, frequency domain optical coherence tomography (FD-OCT) was used for their analysis. Resin restored tooth was prepared. For morphological observation, serially sectioned teeth, conventional X-ray taking and micro computed-tomography (CT) images were compared with OCT images. The experiment has done to evaluate the success of the resin restoration using 3 dimensional structure OCT image. In this research, OCT is evaluated as a new technique to image resin restoration. The evaluation of resin restored tooth was performed by OCT. Inappropriate restoration such as marginal adaptation, large porosities, internal integrity and poor contour could be detected. Resin filler also could be checked by OCT. The distribution, number, regularity and size of resin filler can be differentiated from several commercial products. Considering the characteristics of the OCT, it can be used to evaluate the defects of resin restoration, resin filler distribution, and internal integrity between resin material and tooth structure. The OCT can be considered to be a new and advanced method for the evaluation of resin restorations.

The Chinese ecosystem (especially in traditional markets) has been completely subverted by on-line stores such as Taobao and JingDong after 2010. This research is to understand how to improve traditional markets since 92.5% practitioners in the retail industry conduct business in traditional markets. By the mean time Korean traditional markets attract more customers. In this research the multi-group analysis is used to analyze the difference between Chines customers and Korean customers upon customer equity in traditional markets. The research objectives are: First, to understand the relationships among service quality, customer equity drivers and customer satisfaction in Chinese and Korean traditional markets; Second, find out how to improve the three drivers of customer equity through these variables mentioned above in traditional markets. In this study the relationships among service quality, the drivers of customer equity, customer satisfaction and customer lifetime value were studied based on the analysis of the data which were collected in Chinese and Korean traditional markets.

The Korea Mushroom Resource Bank (KMRB) was launched as a national research resource bank in 2015 by the Ministry of Science, ICT and Future Planning. The main goal of the KMRB is to secure important biological resources, mushroom-forming basidiomycota, significant sources of fundamental and novel substances and materials, as dried specimen, cultures, and genomic DNA. For wider application of fungal resources in education, medicinal and industrial uses, the KMRB will undertake following tasks: 1) Survey natural environments across Korea to catalogue mushroom diversity, 2) Establish resource management system based on accurate identification of mushroom, 3) Evaluate the usefulness of the discovered mushroom, 4) Create a secure preservation and loan system. With a global focus on utilizing natural resources, mushroom resources provide excellent opportunities for academic research, and discovering novel substances for use as medicine and energy.

In Korea, six mealybug species have been reported on pears. This study investigated the occurrence of mealybugs in 19 pear orchards from 2013 to 2014. Two species, Crisicoccus matsumotoi (Siraiwa) (Hemiptera: Pseudococcidae) and Pseudococcus comstocki (Kuwana) were mainly found. The dominant species was C. matsumotoi identified on 73% of infested fruit. Toxicities of 14 registered insecticides (7 single formulations and 7 mixed formulations) commonly used to control P. comstocki were evaluated to C. matsumotoi nymphs and adults at the recommended concentration. As a results, all insecticides exhibited strong insecticidal activity with 100% mortality of both nymphs and adults. These results indicate that the 14 insecticides can be used in control for C. matsumotoi in field.

The Korea Mushroom Resource Bank (KMRB) was launched as a national research resource bank in 2015 by the Ministry of Science, ICT and Future Planning. The main goal of the KMRB is to secure important biological resources, mushroom-forming basidiomycota, significant sources of fundamental and novel substances and materials, as dried specimen, cultures, and genomic DNA. For wider application of fungal resources in education, medicinal and industrial uses, the KMRB will undertake following tasks: 1) Survey natural environments across Korea to catalogue mushroom diversity, 2) Establish resource management system based on accurate identification of mushroom, 3) Evaluate the usefulness of the discovered mushroom, 4) Create a secure preservation and loan system. With a global focus on utilizing natural resources, mushroom resources provide excellent opportunities for academic research, and discovering novel substances for use as medicine and energy. PⅣ-2

Hypocreales entomopathogenic Beauveria bassiana and Metarhizium anisopliae are well-known biological control agents worldwide and have high potential in industrialization. However their thermo-susceptibility limits long-term storage under high temperature conditions and high insecticidal activity after application to target pests. Herein we isolated highly virulent isolates, B. bassiana JEF006 and JEF007 and M. anisopliae JEF003 and 004, and produced in three grains, as substrates for solid cultures, followed by thermotolerance assays to compare the potential of the three substrates. The JEF isolates were exposed to dry and wet heat at 50°C and overall conidia were more stable under dry heat condition rather than wet heat. Of the three grains, Italian millet was superior to the other grains in the production of thermotolerant conidia. Additionally Italian millet did not severely aggregated, which enabled air to penetrate into the substrate well compared to the sorghum and millet. JEF isolates were more thermotolerant when they were kept in oil conditions. The conidia of JEF 007 and 003 wild type and AtMT-based generating random mutants were subjected to SDS-PAGE. A significant relationship between conidial thermotoelrance and detected proteins was observed. This work suggests that Italian millet can be used as an effective substrate to produce more thermotolerant conidia, thus maintaining viability for long times under unfavorable environment and biological activity against target pests.

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.

Entomopathogenic fungi are facultative microorganisms, dwelling in soil or infecting host insects, and some of the genera have been used as biological control agents worldwide. Collection of fungal isolates should be a platform for the development of highly effective resources, thus in this work we constructed a fungal library using a mealworm pathogenecity-based fungal collection method and further characterized some isolates with high virulence. A phylogenetic three was generated, and of the isolates 17 isolates’ biological features were characterized, such as morphology, spectrum of virulence, cultural characteristics, thermo-stability of fungi, production of biologically active materials, such as enzymes. This work reports an attractive entomopathogenic fungal library including the information of effective isolates in pest management.

The present issue of insect resistance and environmental toxicity of pesticides is triggering deep discussion about the pest management tactics, in which pest monitoring and control activity are mainly involved. Novel control agents, hopefully overcoming the present issues and problems, should be researched and commercially applied to the farm fields. With the monitoring-based research, additionally we have to focus on the control-based, particularly control agent-based research and application. Entomo- pathogenic fungi can used as one of the possible novel control agents once considerations are given to the control of soil- or water-dwelling pests. In our research group, the entomopathogenic fungal library has been constructed using the mealworm-based isolation system, which showed a variety of opportunities of their use in pest control. Important key production technologies including granular formulation have been developed to increase their industrialization. Some entomopathogenic fungal isolates showed high biological performance in the control of rice weevils, western flower thrips and Japanese bettles in field stands. To elucidate the fungal mode of action, a fungal transformation system using AtMT and gene identification tools were established. Recently a more deep study about the relationship between insect and entomopathogenic fungi is be investigated using RNA seq. We suggest that to make the entomopathogenic fungal products be applied to agricultural farm field, R&D of down-stream process should be seriously considered as the key step.