CDK2 inhibition plays a central role in DNA damage–induced cell cycle arrest and DNA repair. However, whether CDK2 also influences early porcine embryo development is unknown. In this study, we examined whether CDK2 is involved in the regulation of oocyte meiosis and early embryonic development of porcine. We found that disrupting CDK2 activity with RNAi or an inhibitor did not affect meiotic resumption or MII arrest. However, CDK2 inhibitor-treated embryos showed delayed cleavage and ceased development before the blastocyst stage. Disrupting CDK2 activity is able to induce sustained DNA damage as demonstrated by the formation of distinct γH2AX foci in nuclei of day 3- and day 5-embryos. Inhibiting CDK2 triggers a DNA damage checkpoint by activating of the ATM-P53-P21 pathway. However, the mRNA expression of genes involved in non-homologous end-joining (NHEJ) or homologous recombination (HR) pathways for double strand break (DSB) repair reduced after administering CDK2 inhibitor to 5-day-old embryos. Furthermore, CDK2 inhibition caused apoptosis in day 7 blastocysts. Thus, our results indicate that an ATM-P53-P21 DNA damage checkpoint is intact in the absence of CDK2; however, CDK2 is important for proper repair of the damaged DNA by either directly or indirectly influencing DNA repair-related gene expression.

Mitotic spindle formation is regulated by centrosomes, composed of a centriole pair surrounded by pericentriolar materials(PCM) proteins. However, mammalian oocytes rely on acentriolar MTOCs for the function of meiotic spindle. The composition of acentriolar MTOCs and the molecular precesses that regulate the localization and accumulation in mammalian oocyte are not well understood. In this study, we analyzed the mechanisms of spindle microtubule nucleation and stability from MTOCs in mouse oocyte, and indentified Centrosomal protein192(CEP192) as a key regulator for acentriolar MTOC formation. CEP192 specifically colocalized with pericentrin (PCNT) during the oocyte maturaion. CEP192 proteins are localized throughout cytoplasm and around nucleus at GV stage, and then after BD stage, CEP192 proteins were further fragmented into smaller MTOCs around chromosomes. At metaphase, CEP192 proteins were concentrated in spindle pole. Knockdown of CEP192 using siRNAs resulted in metaphase I arrest. The arrested oocytes were characterized by reduced microtubule intensity and misalignment chromosome. Also at BD and ProMI stage, the oocytes reduced microtubule density and PCNT intensity. To confirm the mechanism of CEP192 regulation, we confirmed that PLK1 and AuroraA kinase were involved in CEP192 activation. The investigations for detailed molecular mechanisms of CEP192 and RanGTP for microtubule nucleation in oocytes are underway using various techniques including siRNA, mRNA, and positive or negative dominant injection and inhibitors.

Prolonged communication between oocytes and the surrounding somatic cells is one of the unique reproductive physiology in canine. Paracrine Kit ligand (KITL) signaling is a well-known communication between granulosa cells and the oocyte. KITL is a cytokine growth factor secreted by granulosa cells that signals via the c-kit receptor expressed by oocytes. Paracrine factors, including growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), exert their effects by binding with the kinase receptors expressed on the granulosa cells. However, the regulations of GDF9 and BMP15 in the canine KITL expression are currently poorly understood. Therefore, we investigated the effects of GDF9 and BMP15 on the expression of KITL in canine ovarian granulosa cells in vitro. In Annexin V assay recombinant GDF9 and BMP15 did not induce apoptosis in the cultured ovarian granulosa cells. When treated, FSH significantly increased KITL expression, and hCG suppressed its expression. When both FSH and hCG were treated, the expression of KITL was affected by GDF9 and BMP15 in dose and time dependent manner in the luteal granulosa cells. GDF9 (10 ng/mL) significantly decreased KITL expression after12 h. BMP15 (10 ng/mL) significantly also decreased KITL expression after 24 h. Western blot and immunochemistry results indicate that GDF9 activated Smad2/3. After blocking ALK 4/5/7 receptors by SB, GDF9 failed to activate Smad2/3, also BMP15 did not activate Smad1/5/8 after blocking ALK 2/3/6 receptors by DM. So GDF9 exerts its effects via using ALK 4/5/7 receptors to activate SMAD2/3 signaling, and BMP15 binds ALK 2/3/6 receptors to activate SMAD1/5/8 signaling. The expression of KITL was not changed by SB or DM treatment. However, the effect of GDF9 and BMP15, which decreased the expression of KITL, was suppressed by SB or DM treatment. In conclusion, this study provides the first evidence that recombinant GDF9 and BMP15 decrease KITL expression in canine ovarian granulosa cells.

The cancer and Parkinson's disease associated protein DJ-1 is multifunctional protein that involves in diverse cellular process. DJ-1 protein has a cellular protective role and promoted cell survival under an oxidative stress. However, the cellular protective mechanism of DJ-1 is not fully understand, and we needs to be further study their functions in novel organisms. In the present study, we investigated the protective role of DJ-1 against induced oxidative stress in canine cell line. On the basis of these experiments, canine DJ-1 overexpressing and null cell lines were established. The stable overexpression and down regulation of DJ-1 efficiency confirmed by the western blot analysis. Subsequently, the DJ-1 gene transfected cell lines and control cells were subjected to induced the oxidative stress, and then cell viability, cell proliferation assay, cellular apoptosis detection analysis (Annexin V and TUNEL assay), intracellular ROS and mitochondrial activity were measured appropriately. The results showed that DJ-1 overexpressed cells were up-regulated cell viability under oxidative stress conditions induced by the rotenone and hydrogen peroxide (H2O2), whereas loss of DJ-1 cells were down-regulated the cell survival activity. Additionally, overexpression of DJ-1 cells increased cell resistance to oxidative stress and inhibited the elevation of cell death and cellular ROS induced apoptosis. Moreover, DJ-1 overexpressed cells was increased mitochondrial functions by using confocal microscopy with MitoTracker staining. On the contrary to this, DJ-1 null cells show defective cellular protection and mitochondria activity against oxidative stress conditions. Our data indicate that canine DJ-1 protein attenuates cellular apoptosis and ROS generation, enhances the cellular survival activity and promote mitochondrial function under the oxidative stress, likewise other mammalian cells. Importantly, DJ-1 overexpression may be an important part of a protective strategy as a sensor for oxidative stress.

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.

Beauveria bassiana JEF-007 with strong virulence against Riptortus pedestiris was selected for the Agrobacerium tumefaciens-mediated transformation(AtMT). AtMT generated two transformants, B1-06 and C1-49, showed significantly reduced virulence against R. pedestris. To identify the virulence-related genes, thermal asymmetric interlaced(TAIL) PCR and flanking region analysis were performed. From the analysis, two genes, Complex I intermediated-associated protein 30(CIA30) and Autophagy protein 22(Atg22), possibly related virulence in B. bassiana JEF-007. For the analysis of two putative virulence-related genes in JEF-007, hairpin RNA interference (hpRNAi) is under consideration. This work can provide the functional roles of the virulence-related genes in 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.

Chemical pesticides have been used to control persimmon pests, however the overuse of the pesticides caused insect resistance. Herein we investigate the potential of eco-friendly organic pesticides (EFOP) on the control persimmon pests, Stathmopoda masinissa and Riptortus pedestris. Eleven commercially available EFOP were sprayed on the target insects in laboratory conditions. The chemical pesticide, buprofezin+dinotefuran wettable powder served as a positive control. In the bioassay against persimmon fruit moth, the mixture of Chinese scholar tree, goosefoot and subtripinnata extracts (EFOP-2) treatment showed the highest control efficacy: 27.7% (5 days) of survival rates. In the bioassay against bean bugs, the EFOP-2 showed the highest control efficacy: 20.0% (5 days) of survival rates. These results suggest that the mixture of three plants extracts (EFOP-2) has high and multiple potential in the management of the persimmon pests.

The spotted-wing drosophila (SWD), Drosophila suzukii (Diptera: Drosophilidae), is an economically damaging pest that feeds on most thin-skinned fruits. In this study, we sequenced portions of the mitochondrial (mt) COI and ND4 genes from a total of 195 individuals collected mainly from Korea. A total of 139 haplotypes were obtained from the concatenated COI and ND4 sequences. A dataset combining GenBank sequences with our own data identified a total of 94 worldwide COI haplotypes with a maximum sequence divergence of 5.433% (32 bp). A rough estimate of genetic diversity in each country showed higher diversity in ancestral distributional ranges, but the invasion over Asian countries seems to have been substantial because haplotype diversity was only 2.35-3.97-fold lower in the USA, Canada, and Italy than that in the populations ancestral ranges.

Alternative control agents, hopefully overcoming the present issues and problems in agrochemicals, should be considered and finally applied to the vector controls. Entomopathogenic fungi can used as one of the possible novel control agents and great considerations are given to the control of soil- or water-dwelling stages of vectors, such as mosquitoes and ticks. 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 vector 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 mosquitoes and ticks in field stands. R&D of down-stream process should be seriously considered as the key step.

Choline is an amine essential to maintenance of good health. Here, we modified the ion chromatographic method for measuring choline described by Laikhtman & Rohrer and validated by the ICH guideline and applied it to monitoring of choline content in infant formula in the Republic of Korea. The Certified Reference Materials (CRM), NIST 1849a, were used for validation of the modified method. The parameters of validation were linearity, range, specificity, accuracy, precision, LOD, LOQ and recovery. The linearity, R2, was above 0.999 and the analytical range was 2.5 to 50 μg/mL. The specificity was confirmed by the retention time (RT) and positive tests with calibration standard and CRM. The accuracy, precision and recovery values were 99.6 ± 2.41 %, 2.42 % and 90.8 - 92.8 %. The LOD and LOQ values were 0.03 mg/kg and 0.17 mg/kg, respectively. Five livestock products inspection agencies voluntarily participated in the collaborative study. The result, HORRAT value was 0.8 which was within the reference value 0.5 – 2.0. The validated method was applied to the analysis of choline in infant formula in the Republic of Korea. The results also conformed to 'The Standard for Livestock Products Labeling'.

Vitamin K1 is a fat-soluble vitamin essential for human health. We used the Association of Official Analytical Chemists(AOAC) method 999.15, validated by the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use(ICH) Guideline, to monitor vitamin K1 content in infant formula in the Republic of Korea. The Certified Reference Materials (CRM), NIST 1849a, were used for validation of the test method. The parameters of validation were linearity, range, specificity, accuracy, precision, LOD, LOQ and recovery. The linearity, R2 was above 0.999 and the analytical range was 0.001 to 0.22 μg/mL. The specificity was confirmed by the retention time (RT) and positive/negative sample test. The accuracy, precision and recovery values were 101.13 ± 0.13 %, 3.15 % and 81.60 % - 99.78 %. The LOD and LOQ values were 0.582 ng/mL and 1.76 ng/mL, respectively. Five livestock products inspection agencies voluntarily participated in the collaborative study. The HORRAT value was 0.6, which was within the reference value 0.5 – 2.0. The validated test method was applied for the monitoring of vitamin K1 in infant formula in the Republic of Korea. The results also conformed to 'The Standard for Livestock Products of Processing and Ingredient Specifications' and 'The Standard for Livestock Products Labeling'.

Acorus gramineus is a herb used frequently in oriental medicine. Clinically, the effects of Acorus gramineus are known as awakening of one’s consciousness, removal of sputum, and relief of autism. Experimentally, Acorus gramineus root(AGR)-induced cerebral blood flow increase depending on the concentration causes diverse responses of blood pressure and regional cerebral blood flow and possesses strong inhibitory effects of apoptosis in the nervous system. We investigated the effects of AGR water extract on the adipose tissue status. Male C57BL/6 mice were fed high fat diet and a total of 35 mice were divided into 5 groups: normal group, control group, and groups treated with AGR water extract at concentrations of 20, 100, and 500 mg/kg. The mice were treated by oral administration once a day for consecutive 6 weeks, and their weight changes were monitored. As the results, the groups treated with AGR water extract showed weight loss. Also, the weights of liver, epididymal fat, retroperitoneal fat and peri-renal fat were decreased in the groups treated with AGR water extract. In histopathological examination, the sizes of adipose tissues decreased in liver, epididymal fat, retroperitoneal fat, and peri-renal fat. These results indicate that AGR water extract have an anti-obesity activity, which could be used as an useful material to decrease body adipose tissues.

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.

Supply and demand patterns have dynamically changed in recent years due to the increased global competition. Firms have selected quality and price as competition components and have accomplished corporate innovation in order to achieve competitive advantage. Innovation has been recognized as the way to enhance corporate competitiveness and to continuously grow in churning global competition (Homburg, Schwemmle, & Kuehnl, 2015; Moon, Miller, & Kim, 2013). Although innovation becomes a common means to improve a firm’s performance, it has the limitation of achieving a firm’s strategic goal as a long-term strategy. Thus, firms need to have more fine-grained strategies to survive in dynamically changing business environment, such as design innovation (Moon et al., 2015). For example, Apple has produced its products (e.g., iPhone, iPod and iPad) focusing on innovative product design to influence consumer purchase intention. Design can lead to a distinct competitive advantage (Bolch, 1995). Furthermore, product design can be used by firms to create amd enhance brand recognition, as well as to increase firms’ value (Mozota, 2002). Brand experience positively influences customer satisfaction and brand loyalty (Brakus, Schmitt, & Zarantonello, 2009). Thus, we assume that design experience and product innovation have a positive effect on consumers’ purchase behavior and customer value. However, while the importance of design innovation is recognized, it is not easy to apply the design innovation to marketing due to the lack of relevant research in the field. In fact, relevant research on the influence of innovative new product design and design experience on customer value is scarce. Therefore, the purpose of this study is to investigate how brand experiences and innovative product design affect customer value. Successful innovation is accomplished by identifying customer needs first and developing innovative products to satisfy their needs (Hauser et al., 2006). Design is seen as the core of innovation and the moment when a new object is conceived of, devised, and shaped in a prototype form (Landwehr, Wentzel, & Herrmann, 2013). Verganti (2008) studies the concept of "user-centered design," which describes how companies can use design to improve their relationships with users and develop a better understanding of user needs. In this research, design innovation has three dimensions: aesthetic attributes, feature attributes, and emotional attributes. First, aesthetic attributes focus on the product design itself. The aesthetic appearance of a product has a large bearing on its potential market share (Liu, 2003). Second, feature attributes focus on the product features and functional aspects that are required to satisfy customer needs. Feature attributes enable performance that can give results in the operating process (Crawford & Di Benedetto, 2007). Third, emotional attributes focus on consumers’ feeling when they purchase a new product to satisfy their needs. Emotional attributes are generated by consumers’ experience when they purchase a new product in the store. The more the product design satisfies customer's emotional needs, the more customers’ attention is attracted to the purchase of a product (Mokarian, 2007). A product satisfying the aesthetic, feature, and emotional attributes through design innovation provides a new experience to customers (Desmet & Hekkert, 2007). A consumer’s purchase decision making is affected by both direct and indirect experience of using the product and the function of product (Brakus, Schmitt, & Zarantonello, 2009). Therefore, a designer comes to design the product, taking an interest in the experience that the product gives besides its shape and function. Product experiences occur when a customer interacts with the product ̶ for example, when customers search for, examine, and evaluate products (Hoch, 2002). The product experience can be direct, i.e. when there is physical contact with the product (Hoch & Ha 1986) or indirect, i.e. when a product is presented virtually or in an advertisement (Hoch & Ha 1986; Kempf and Smith 1998). Brand experience can be split into four dimensions (sensory, affective, intellectual, and behavioral), which are differentially evoked by various brands (Brakus, Schmitt, & Zarantonello, 2009). According to previous studies, these four experience dimensions (sensory, affective, intellectual, and behavioral experience dimensions) have an effect on customer satisfaction and customer loyalty. Customer value can be defined as the trade-off between the benefits and sacrifices as a customer purchases a product or service from a supplier (Ulaga & Chacour, 2001). Ulaga and Chacour presented three dimensions of customer value: product-related components, service-related components, and promotion-related components. The researchers argued that customer value could be an important strategic marketing tool to clarify a firm's proposition to customers. Product-related components are intrinsic product characteristics. Product quality is a key factor of relationship value (Ulaga, 2003). Customer value consists of product value and service value. Customer value can be enhanced by quality, diversity, payment, and service quality and decreased by price, convenience, and risk (Jarvenpaa & Todd, 2003). Service-related components include all aspects of service associated with the product. Various service components play an important role in differentiating a supplier's offering (Narus & Anderson, 1996). Promotion-related components include all items used to promote the product to the customer. For all purchasing processes, it is necessary to assess the perceived customer value, such as service quality and promotional quality (Qualls & Rosa, 1995). In the present study, we conducted a research survey with 300 subjects and analyzed the data. In order to test the reliability of questionnaire, Cronbach's alpha was used. In order to test the validity, an exploratory factor analysis was conducted. Measurement was made for 3 variables, i.e. design innovation, design experience, and customer value. Three variables (aesthetical attributes, feature attributes, and emotional attributes) were used for design innovation. Three types of attributes were introduced for a mobile phone design innovation. Four dimensions (sensory, affective, intellectual, behavioral) were measured using a technical design experience and a humanistic design experience. Three variables (product-related customer value, service-related customer value, and promotion-related customer value) were employed for customer value. The results of the analyses demonstrate that design innovation has a positive effect on design experience, while design experience has a positive effect on customer value. Aesthetic attributes of design innovation have a positive effect on technical design experience and humanistic design experience. Feature attributes have a positive effect on the technical design experience and the humanistic design experience. Emotional attributes have an effect on the technical design experience and the humanistic design experience. The technical design experience has a positive effect on product-related customer value, service-related customer value, and promotion-related customer value. The humanistic design experience has a positive effect on product-related customer value, service-related customer value and promotion-related customer value. This study will make it possible to empirically examine how customer's experience in design innovation affects customer value. Our results will provide a theoretical foundation for examining a relationship between variables regarding how design innovation influences customer value through design experience. It is intended to give a direction as to the design innovation of firms by clarifying and presenting antecedent factors having an effect that design innovation produces on customer value. The results of the present study will inspire designers to design in consideration of design experience. Finally, our study will provide marketers with guidelines as to how design experience can influence customer satisfaction and loyalty.