Since rice is the main food in Korea, there are no regulations on corn milling yet. Corn is known as one of the world's top three food crops along with wheat and rice, and it is known that 3.5 billion people worldwide use corn for food. In addition, corn mills are not developed or sold in Korea, but the use of corn mills is increasing significantly in many countries in Southeast Asia. In the Philippines, as Korea's rice mill import increases, Korea's KAMICO (Korea Agricultural Machinery Industry Cooperative) and domestic company A agreed to develop a corn mill jointly with PHilMech, an organization affiliated with the Philippine Ministry of Agriculture. However, research on corn milling was very insignificant, so the development was carried out based on the technology of Korea's rice mill. Rice milling is performed by peeling off the skin of rice and producing brown or white rice, so it is carried out by removing the skin and cutting the skin. On the other hand, in the corn mill, the skin of the corn is peeled, pulverized and selected to produce main products suitable for edible use. Therefore, in order to develop a corn mill, processes such as peeling, transfer, grinding, sorting, and by-product separation are required, and suitable parts must be developed. In addition, the performance must be gradually improved through experiments in which corn is repeatedly milled. The Philippines produces 7.98 million tons/year of corn, which is about 100 times that of Korea, and is mostly consumed as a staple food. This is about 10% of the total crop production in the Philippines. In addition, the main cultivation complexes of corn are the mountainous regions of Tarlac or Pangasinan, and the produced corn is 72.4% of the so-called yellow corn called Arabel and Sarangani, and the remaining 27.6% are known as white corn. In this study, it was intended to produce grains of 2.5 mm or less suitable for food for yellow corn and to develop a corn mill for 200 kg per hour. Detailed conditions for development are stipulated as more than 55% of the main product recovery rate, more than 31% of the by-product recovery rate, less than 5% of the raw material loss rate, and more than 80% of the embryo dislocation rate. In this study, to achieve this, the overall process of the corn mill was developed, and the optimal conditions for the corn mill were obtained through the development of parts and empirical tests to improve performance. In addition, it was intended to achieve the development goal by evaluating and analyzing the performance of each part so that it did not conflict.

Bees of the genus Osmia are cavity-nesting solitary species that play important roles as pollinators. Nest traps are one of the most common methods to study Osmia bees. To elucidate the optimal environmental conditions of nesting sites, we investigated the effects of location, direction, altitude, and sites of nest traps on the rate of trap-nesting Osmia spp. During the collection period, the average rate of trap-nesting Osmia spp. collected within 90 days after the installation of traps was 17.0±20.0%. This percentage was 2.7-fold higher than that of trap-nesting bees in 30 days after the installation of traps. The Jeongseon location exhibited the highest rates of trap-nesting Osmia spp. collected in 30 and 90 days, representing 11.1 ±17.6% and 23.2±22.5%, respectively. The direction of the nest traps did not affect the rate of trap-nesting Osmia spp. The altitude ranges of the traps were 0-199 m, 200-399 m, 400-599 m, 600-799 m and over 800 m. Interestingly, the altitude range of 600-799 m showed the highest rate of trap-nesting bees, which was 40.4±3.9%. Higher altitudes seemed to correspond to higher rates of trap-nesting bees. With regards to the sites where the nests were placed, the rate of trap-nesting bees in a mud wall of an old house was 45.1±25.2%, which was 3-fold higher than that of a nest in an apple orchard. The flowering plants collected at different locations during the trap-nesting activity of Osmia bees belonged to 18 families and 34 species. In conclusion, the rates of trap nests colonized by Osmia spp. were affected by altitude, site, and plant diversity.

Polyhedrin is the major component of the nuclear viral occlusions produced during replication of the baculovirus Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV). To enhance the production efficiency of foreign protein in baculovirus expression system, the effects of various polyhedrin fragments were investigated by fusion expressing them with the enhanced green fluorescent protein (EGFP). Recombinant viruses were generated to express EGFP fused with polyhedrin fragments based on the previously reported minimal region for self-assembly and the KRKK nuclear localization signal (NLS). The marked increase of EGFP by these fusion expressions was confirmed through protein and fluorescence intensity analyses. Among the fusion-expressed protein in nucleus and cytoplasm, the most hyper-expression was observed in the fusion of amino acids 19 to 110 and 32 to 59 of polyhedrin. Also these fragments, some degradation of only the fused polyhedrin was observed in the fusion of amino acids 19 to 85 and 32 to 85. The production of E2 protein, which is a major antigen of classical swine fever virus, was dramatically increased by fusion expression with polyhedrin amino acids 19 to 110, and its preliminary immunogenicity was verified using experimental guinea pigs. The production of luciferase was approximately two folds increased by fusion expression with polyhedrin amino acids 32 to 59, and its activity was measured using Luminometer. This study suggests a new option for higher expression of useful foreign recombinant protein using the partial polyhedrin fusion expression in baculovirus.

The fact that flip-flops, one of many different types of unstable shoes, are light and relatively easy to put on, accounts for their popularity among people. But because flip-flops rely heavily on the support of a single thong between your first and second toes, they impose a huge amount of pressure onto lower leg. Thus in the following experiment we tried to examine the different effects of flip-flops and running shoes in terms of their effect on muscle activity and fatigue of tibialis anterior and gastrocnemius during walking. In order to measure an electromyogram we used Free EMG system. 10 men and 10 women in running shoes ran on treadmills for 15 minutes at 4.8km/h, 2 days later the same experiment was carried out, but this time, in flip-flops. p value turned out to be greater than .05 and thus there was no considerable difference between the effects of flip-flops and running shoes on muscle activity and fatigue during walking. Therefore we conclude that despite the fact that flip-flops are considered unstable, their effects on muscle activity and fatigue of tibialis anterior and gastrocnemius are negligible.

Insect-killing fungi have high potential for controlling agriculturally harmful pests. However, their pathogenicity is slow and this is one reason for their poor acceptance as a fungal insecticide. The expression of bumblebee, Bombus ignitus, venom serine protease (VSP) by Beauveria bassiana ERL1170 induced melanization of yellow spotted longicorn beetles, Psacothea hilaris as an over-reactive immune response, and caused substantially earlier mortality in beet armyworm, Spodopetra exigua larvae when compared to the wild type. No fungal outgrowth or sporulation was observed on the melanized insects, thus suggesting a self-restriction of the dispersal of the genetically modified fungus in the environment. The research is the first use of a multi-functional bumblebee VSP to significantly increase the speed of fungal pathogenicity, while minimizing the dispersal of the fungal transformant in the environment

ORF78 (ac78) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a baculovirus core gene of unknown function. To determine the role of ac78 in baculovirus life cycle, an ac78-deleted mutant AcMNPV, Ac78KO, was constructed. Quantitative PCR analysis revealed that ac78 is a late gene in the viral life cycle. After transfection into Spodoptera frugiperda cells, Ac78KO produced a single-cell infection phenotype indicating that no infectious budded viruses (BVs) were produced. The defection in BV production was also confirmed by both viral titration and Western blot. However, viral DNA replication is unaffected. Analysis of BV and occlusion derived virus (ODV) revealed that AC78 is associated with both forms of the virions and is a structural protein located to viral envelope. Electron microscopy showed that ac78 also plays an important role in embedding of ODV into occlusion body. This study therefore demonstrates that AC78 is a late virion associated protein and is essential for the viral life cycle.

ORF11 (ac11) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a highly conserved gene of unknown function. To determine the role of ac11 in baculovirus life cycle, an ac11-knockout mutant AcMNPV, Ac11KO, was constructed. qPCR analysis revealed that ac11 is an early gene in the life cycle. After transfection into Spodoptera frugiperda cells, Ac11KO produced a single cell infection phenotype indicating that no infectious budded viruses (BVs) were produced. The defection in BV production was confirmed by both viral titration and Western blot. However, viral DNA replication is unaffected. Electron microscopy showed that ac11 is required for nucleocapsids envelopment to form ODV and their subsequent embedding into OB. This study therefore demonstrates that ac11 is an early gene which is essential for the viral life cycle.

To enhance the production efficiency of foreign protein in baculovirus expression system, the effects of polyhedrin fragments were investigated by fusion expression them with the enhanced green fluorescence protein (EGFP). Recombinant viruses were generated to express EGFP fused with polyhedrin fragments based on the minimal region for self-assembly and the KRKK nuclear localization signal (NLS). The increase of EGFP production by fusion expressions was confirmed through protein and fluorescence intensity analyses. The importance of nuclear localization for enhanced production of EGFP was shown by the mutation of the NLS within the fused polyhedrin fragment. Among the fusion expressed protein in cytoplasm, the most hyper-expression was observed in the fusion of amino acids 32 to 59 of polyhedrin. Polyhedrin fragment fusion expression with classical swine fever virus E2 protein also resulted hyper-enhanced expression of E2 protein. However, the fusion expression of porcine circovirus ORF2 with polyhedrin fragment did not show significant enhance of ORF2 production. These results suggested that the enhancement of foreign protein production when fused with polyhedrin is caused by the enhanced stability of expressed protein.

This study was conducted to evaluate the effect of the exercise on elderly balance ability by using hippotherapy and therapeutic ball exercise. 10 patients were assigned to the hippotherapy group and they got with 30 minutes of hippotherapy. Another 10 elderly were assigned to the therapeutic ball group and they got with 30 minutes of therapeutic ball exercise. All procedures were repeated 5 times a week for the total of four weeks. To investigate the participants balancing abilities, the Time“ Up & Go”(TUG) and One Leg Stand Test(OLST) were evaluated. The results of study were significant differences between pre-test and post-test of TUG and OLST(p<.05), and there were no significant differences between hippotherapy and therapeutic ball exercise(p>.05). The conclusion showed that both the hippotherapy and the therapeutic ball exercises were effective on elderly balancing ability. Consequently, it would be better to practice therapeutic ball than hippotherapy for elderly exercise because the more economical and there is less restriction of space than the hippotherapy.

Insect-killing (entomopathogenic) fungi have high potential for controlling agriculturally harmful pests. However, their pathogenicity is slow and this is one reason for their poor acceptance as a fungal insecticide. The expression of bumblebee, Bombus ignitus, venom serine protease (VSP) by Beauveria bassiana (ERL1170) induced melanization of yellow spotted longicorn beetles (Psacothea hilaris) as an over-reactive immune response, and caused substantially earlier mortality in beet armyworm (Spodopetra exigua) larvae when compared to the wild type. No fungal outgrowth or sporulation was observed on the melanized insects, thus suggesting a self-restriction of the dispersal of the genetically modified fungus in the environment. The fungal transformant also shows mammal fibrinolytic activity, by which the transformant can be used pharmaceutically. The research is the first use of a multi-functional bumblebee VSP to significantly increase the speed of fungal pathogenicity, while minimizing the dispersal of the fungal transformant in the environment.

Palatal development is one of the crucial events in craniofacial morphogenesis, according to the significant signaling pathway including the out growth, elevation, and fusion of palatal shelves. In the fusion of palatal shelves, epithelial to mesenchymal transition (EMT) is a fundamental process to achieve the proper morphogenesis of palate. Mechanisms of EMT have been reported as the processes of migration, apoptosis or general EMT through the modulations through various signalling molecules. Rgs19, known as a regulator of G protein signaling (RGS) family through GTPase activity, showed the interesting epithelial expression patterns in various organogeneses including the significant expression patterns of Rgs19 in palatal development. To evaluate the precise function of Rgs19 in palatogenesis, we employed the gain and loss of function studies using ASODN treatments and gene electroporations while in vitro palate organ cultivations. Knockdown of Rgs19 using treatments of AS-ODN showed the retarded palatal fusion with the decreased patterns of apoptosis in mesial epithelium edge (MEE). In addition, alteration patterns of related genes were examined with the qRT-PCR. And epithelial mesenchyme transition (EMT) process was delayed in medial edge epithelium (MEE) throught immunohistochemistry of pancytokeratin, which known as epithelial cell marker. Morphological changes were observed with the three dimensional reconstruction method. These results show that expression of Rgs19 in MEE has crucial role of EMT, also Rgs19 affects to palatal fusion by regulation of apoptosis through the signalling modulations.

ORF11 (ac11) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a highly conserved baculovirus gene whose homologs are found in all lepidoteran Group I NPV, but its function is unknown so far. To determine the role of ac11 in baculovirus life cycle, ac11 knock-out mutant, Ac11KO, was constructed using the plasmid capture system (PCS). Real-Time PCR analysis showed that ac11 transcript was first detected at 6 h post-infection (p.i.) and accumulated to maximum at 48 h p.i., indicating that ac11 is belong to late gene. When the genomic DNA of Ac11KO was transfected into Sf9 cells, viral replication was restricted to a cell transfected originally. While viral transmission of the Ac11KO was not observed in Sf9 cells, production of budded virus (BV) in Sf9 cells transfected with Ac11KO was observed by transmission electron microscopy (TEM). These results suggest that the ac11 is essential for AcMNPV to produce infective BV.

Palatal development is one of the crucial events in craniofacial morphogenesis, according to the significant signaling pathway. In the fusion of palatal shelves, EMT is a fundamental process to achieve the proper morphogenesis of palate. Mechanisms of EMT have been reported as the processes of migration, apoptosis or general EMT through the modulations through various signalling molecules. Rgs19, known as a RGS family through GTPase activity, showed the interesting epithelial expression patterns in various organogenesis including the significant expression patterns of Rgs19 in palatal development. To evaluate the precise function of Rgs19 in palatogenesis, we employed the loss of function studies using AS-ODN treatments while in vitro palate organ cultivations. Knock-down of Rgs19 using treatments of AS-ODN showed the retarded palatal fusion with the decreased patterns of apoptosis in mesial epithelium edge (MEE). In addition, alteration patterns of related genes were examined with the qRT-PCR. And EMT process was delayed in MEE throught staining of pancytokeratin, which known as epithelial cell marker. These results show that expression of Rgs19 in MEE has crucial role of EMT, also Rgs19 affects to palatal fusion by regulation of apoptosis through the signalling modulations. Palatal development is one of the crucial events in craniofacial morphogenesis, according to the significant signaling pathway. In the fusion of palatal shelves, EMT is a fundamental process to achieve the proper morphogenesis of palate. Mechanisms of EMT have been reported as the processes of migration, apoptosis or general EMT through the modulations through various signalling molecules. Rgs19, known as a RGS family through GTPase activity, showed the interesting epithelial expression patterns in various organogenesis including the significant expression patterns of Rgs19 in palatal development. To evaluate the precise function of Rgs19 in palatogenesis, we employed the loss of function studies using AS-ODN treatments while in vitro palate organ cultivations. Knock-down of Rgs19 using treatments of AS-ODN showed the retarded palatal fusion with the decreased patterns of apoptosis in mesial epithelium edge (MEE). In addition, alteration patterns of related genes were examined with the qRT-PCR. And EMT process was delayed in MEE throught staining of pancytokeratin, which known as epithelial cell marker. These results show that expression of Rgs19 in MEE has crucial role of EMT, also Rgs19 affects to palatal fusion by regulation of apoptosis through the signalling modulations.

Although baculoviruses have a long history of safe use as specific, environmentally benign insect control agents, their use has been limited by several factors, especially their slow speed of action. In this study, we intended to improve the insecticidal activities of Autographa californica nucleopolyhedrovirus (AcMNPV) by expressing Kunitz-type toxin isolated from venoms of Bombus ignitus or Araneus ventricosus. For this, recombinant AcMNPVs, AcBi-KTT, AcAv-Tox1 and AcAv-Tox2 expressing Bi-KTT, Av-Tox1 and Av-Tox2, respectively, under the control of p10 gene promoter were constructed. While polyhedra produced by these recombinant viruses were identical to those of the wild-type AcMNPV in shape, their sizes were relatively smaller than those of the AcMNPV. Among recombinant viruses, AcBi-KTT and AcAv-Tox2 showed significant reduction in median lethal time (LT50) against Spodoptera exigua larvae. Especiaaly, these two viruses showed about 6.2~10-folds higher polyhedra production rate compared to that of the AcMNPV. These results suggested that Kunitz-type toxins from insect venom could be successfully applied to improve insecticidal activity of baculoviruses.

Among 154 putative ORFs of Autographa californica multiple nucleopolyhedrovirus (AcMNPV), ac78 and ac79 are highly conserved genes in baculovirus, but their functions in the virus life cycle have been unknown so far. To determine their roles in AcMNPV replication, knockout mutants, ac78KO and ac79KO, were constructed using the plasmid capture system (PCS). Real-Time PCR analysis showed that both of ac78 and ac79 transcripts were first detected at 6 hours post-infection, and accumulated to maximum at 24 hours post-infection, suggesting that both of ac78 and ac79 are belong to late gene. When the genomic DNA of ac78KO was transfected into Sf9 cells, viral replication was restricted to a single cell infection. These results demonstrated that the ac78 play an important role in BV production, and therefore is essential for AcMNPV to mount a successful infection. Whereas Sf9 cells infected with the ac79KO showed normal viral symptoms such as rounding and swelling, OBs were not observed from majority of infected cells. These results suggested that the ac79 might play an important role in OB production.

Bee venom is a rich source of pharmacologically active substances. In this study, we identified a bumblebee (Bombus ignitus) venom Kunitz-type serine protease inhibitor (Bi-KTI) that acts as a plasmin inhibitor. Bi-KTI showed no detectable inhibitory effect on factor Xa, thrombin, or tPA. However, it strongly inhibited plasmin, although this inhibitory ability was two-fold weaker than that of aprotinin. The activities of B. ignitusvenom serine protease (Bi-VSP) and plasmin in the presence of Bi-KTI indicate that Bi-KTI targets plasmin more specifically than Bi-VSP. These findings demonstrate a novel mechanism for bee venom by which Bi-KTI acts as an antifibrinolytic agent, raising interest in Bi-KTI as a potential clinical agent.

Porcine Circovirus Type2 (PCV2), a single-stranded DNA virus associated with Postweaning multisystemic wasting syndrome(PMWS) of swine, has two major open reading frames, ORF1 and ORF2. The genomic size and molecular weight of ORF2 is respectively 699bp, 28kDa. ORF2 encodes the capsid protein (structural protein) that has type-specific epitopes and is very immunogenic and associated with the induction of neutralizing antibodies, suggesting its potential use in diagnostic assays as well as vaccine development. For efficient production of the capsid proteins, we expressed the PCV2 ORF2 gene with baculovirus in the insect cells. In this study, PCV2 ORF2 was appropriately ligated into the baculovirus transfer vector, pBacPAK9 and pB9-Acpol19-110-EK. Sf21 cells were transfected with a mixture of the purified recombinant transfer vector and bAcGOZA. We generated and purified recombinant viruses containing PCV2 ORF2, and named rAc-B9-PCV2ORF2 and rAc-B9-19-110-EK-PCV2ORF2, respectively. Expression levels of capsid fusion proteins with a partial polyhedrin region of AcNPV more increased than recombinant proteins from non-fusion expressed. Also, expression efficiency increased over time and differed at MOI. As a results, fusion expression of porcine circovirus type2 ORF2 using baculovirus could be utilized as an alternative expression method to produce recombinant antigen against PCV2 infection and is worthy of further investigation.

Polyhedrin is the major component of the nuclear viral occlusions produced during replication of the baculovirus Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV). To enhance the expression level of baculovirus vector system, we constructed several fusion vectors using various fragments of the polyhedrin. The polyhedrin fragments were genetically fused to the enhanced green fluorescent protein (eGFP) under the control of polyhedrin promoter, and their expressions were analyzed in Sf21 insect cells. Expression of the fusion protein was identified by SDS-PAGE and Western blot analysis using anti-GFP and anti-Polyhedrin. The expression level of eGFP was markedly increased by the fusion of partial polyhedrin. Also, the fluorescence intensity of fusion proteins was higher than that of non-fusion protein. Confocal laser scanning microscopy demonstrated that fusion proteins were localized to the cytosol or nucleus of insect cells. In additional, the glycoprotein E2 (gE2) of classical swine fever virus (CSFV) expressed by the these vectors was dramatically increased and its immunogenicity was proofed using experimental animal guinea pigs that were immunized with the partial polyhedrin containing gE2. This study provides a new option for the higher expression of useful foreign recombinant protein by using the partial polyhedrin in BEVS.