West Nile Virus (WNV) is transmitted by infected mosquitoes. Vector mosquitoes usually acquire these pathogens fromfeeding on an infected host, and transmit the pathogens to a naive host during feeding events. To understand the virustransmission dynamics and to survey WNV throughout country, the present study has been conducted. We collected mosquitoesat urban parks in Seongnam, Wonju, Gunsan, Daegu, and Tongyeong using CDC light trap with Dry ice from April toSeptemper in 2017 (mosquito collecting is on going). Among collected mosquitoes, blood-fed mosquitoes were conductedblood meal identification assay and the other mosquitoes were subjected to virus detection using real-time PCR method.A total of 2,290 mosquitoes representing 6 genera and 15 species were collected. The most dominant species was Culexpipiens complex (42.1%) followed by Aedes albopictus (15.1%), Ae. vexans nipponii (14.6%), Ochlerotatus koreicus (9.8%),Cx. orientalis (6.5%), and Armigeres subalbatus (4.4%). The blood meal source were of mammal (93.3%), and birds (6.7%).So far, no WNV has been detected in any mosquitoes.

The members of the genus Flavivirus are noteworthy, as they cause infectious diseases in humans, such as Zika, denguefever, yellow fever, West Nile, and Japanese encephalitis. Due to the increased awareness of the public health risk posedby flavivirus-infected mosquitoes, mosquito collections were performed in six urban parks of South Korea, as the parksare designated for human recreation but also provide suitable habitats for mosquitoes. We examined the diversity andabundance of mosquito species and conducted molecular diagnostics for the detection of flavivirus infections. Monthlycollections were carried out in each park from March to August in 2017. A total of 4,851 mosquitoes (5 genera and13 species) were collected using BG-sentinel traps and then investigated for flavivirus infections. Pathogenic flavivirusinfections causing human diseases were not observed in the field-collected mosquitoes. However, insect-specific flavivirus(ISF) infections were detected in several mosquito pools. ISF has been previously known to enhance or suppress the replicationof medically important flaviviruses in co-infected mosquito cells. In this study, partial sequences of ISF were analyzed.However, further studies are needed in order to determine its genetic characterization and biological function in vivo.

Ochratoxin A, which is frequently detected in cereals and infant diets worldwidely, is a mycotoxin to damage mainly the kidney and liver. Because ochratoxin A is highly thermostable compound. it is necessary to study ways of reducing level of ochratoxin A by controling processing steps. However, food processes, including extrusion, expansion, roasting, and steam cooking, which are used in order to mitigate the contents of ochratoxin A, are known to produce polycyclic aromatic hydrocarbons, which are generated from radicals decomposed by pyrolysis. Therefore, this study analyzed the levels of 4 polycyclic aromatic hydrocarbons, benz (a) anthracene, chrysene, benzo (b) fluoranthene and benzo (a) pyrene in rice-based products made in high pressure and heating process. Rice samples were finely ground, and homogenized samples were alkaline treatement with 1 M KOH/EtOH and extracted with liquid-liquid extraction method using n-hexane. The extracted solution was pretreated with a silica cartridge. The purified solution was dried with nitrogen gas and dissolved in 1 mL of dichloromethane and injected into GC/MSD. We had overall recoveries for 4 polycyclic aromatic hydrocarbons spiked into rice samples ranging from 92.8 to 110.2%. The limit of quantitations of benz (a) anthracene, chrysene, benzo (b) fluoranthene and benzo (a) pyrene in rice-based product were 0.19 ng/g, 0.38 ng/g, 0.51 ng/g, and 0.31 ng/g, respectively. However, these 4 polycyclic aromatic hydrocarbons in all processed rice samples were not detected.

For the development of processed food for hypoglcemia, it is important to construct model system to confirm factors that reduce the glycemic index in real food. This study was to investigate the relationship between the gel model system and real food the high amylose type of segoami using response surface methodology. The independent variables were concentration (X1; 10, 15, 20, 25, and 30%) and steaming time (X2; 30, 35, 40, 45, and 50 min). The predicted glycemic index (Y1) was analyzed as a dependent variable. The regression of pGI was 0.7343, indicating that the model fits the data well in the prediction test. The predict glycemic index of the gel was in the range of 71.38 ~ 83.78, depending on the gel preparation conditions and predicted optimum condition was 23.7% gel concentration and 43.8 minutes of gelatinization. The predict glycemic index of rice gel were decreased with an increase of gel concentration, rather than the steaming time. In order to confirm the effect on in vitro digestibility in real food, the real food (Garraedduk) was prepared by applying the optimal conditions. As a result, there was no significant difference between the predicted value (77.1) and the experiment value (76.6). This result showed that the in vitro digestibility in real food can be predicted by applying the gel model system.

We performed a survey for flavivirus infection and distribution of Aedes albopictus that known as Zika and Dengue virus vector using black–light trap and BG-sentinel trap around urban area in Korea. Mosquitoes were collected in 27 cities during March to November (twice a month) year 2016. Total numbers of mosquitoes collected 102,102 including 19 species 8 genera during collecting period. Total 21,467 Ae. albopictus was collected that 20,961(24.3%) by BG-sentinel trap and 506 (3.2%) by Black-light trap in urban area. Trap index(trap/night) of Ae. albopictus was showed highest in Hamyang (TI:992.3) and lowest in Taebaek (TI:0.3) there was only collected by Black-light trap. A total of 894 pools from all collecting Ae. albopictus were performed a Flavivirus detection. Flavivirus was not detected during study period. This study may provide basic information for surveillance of imported diseases (include Zika virus) and vectors in Korea.

The tobacco cutworm, Spodoptera litura(Fabricius), is a major pest of tomato and frequently demands control measures. The timing of insecticide application is a key factor in determining its efficiency, so an experiment was designed to investigate this. Application of insecticide was based on three criteria: (i) the number of trap-caught moths in a Delta-type trap with a commercial sex pheromone lure placed in the center of the target area, soon after plant emergence; (ii) the percentage of plants exhibiting pinhole-type damage (10% or 20%) and (iii) the percentage of plants exhibiting shot hole-type damage (10% or 20%) compared to a check plot without any control measures. We found that the number of trap-caught moths was, compared to the other methods, the best means of deciding on insecticide application in tomato plant to control the tobacco cutworm. Using pheromone traps, we obtained the best performance of the insecticide Shinnago, causing > 90% larval mortality. Without insecticide application, tomato yield reduction due to the tobacco cutworm larva damage was 27%.

Pheromone biosynthesis activating neuropeptide (PBAN) produced in the subesophageal ganglion is known to stimulate pheromone production in the pheromone gland. A cDNA isolated from female adult heads of Maruca vitrata encodes 197 amino acids including PBAN, designated as Mvi-PBAN, and four other neuropeptides (NPs): diapause hormone (DH) homologue, α-NP, β-NP and γ-NP. All of the peptides are amidated in their C-termini and shared a conserved motif, FXPR(or K)L-NH2 structure. Mvi-PBAN consists of 35 amino acids as previously reported (Chang and Ramasamy, 2014). RT-PCR analysis revealed that Mvi-PBAN cDNA was expressed in all examined body parts. Nucleotide sequence analysis of RT-PCR products indicated the Mvi-PBAN sequence was identical in all examined body parts of both sexes. These results suggest that Mvi-PBAN expression is maintained in examined stages or tissues.

This study was conducted to evaluate the accumulation and distribution of hydrophobically modified glycol chitosan (HGC) as a degradable nanoparticle in the body. To determine the movement of degradable HGC nanoparticles in the body, 20 mg/kg of lutetium177-labeled HGC (Lu177-HGC) with the size ranging from 320 to 400 nm was injected intravenously into ICR mice, and the amount of radioactivity remaining in blood and several organs was measured at various time points during the period of 5 days. In the pharmacokinetics analysis using the Lu177 radioisotope, the free Lu177 was mainly distributed and accumulated in the order of kidney>liver>lung at 1 day after the injection of the radioisotope. However, the Lu177-HGC showed a high distribution of nanoparticles in the order of liver>spleen>kidney during the experimental period of 5 days. These results would provide a basic pharmacokinetics for the use of HGC as a drug carrier in drug delivery system.

The ultimate goal of this study is to assess the accumulation and distribution of hydrophobically modified glycol chitosan (HGC) as a degradable nanoparticle in the body. To understand the movement of degradable nanoparticle HGC in the body, we intravenously injected a dose of 20 mg/kg of Cy5.5-labeled HGC with size ranging from 320 to 400 nm into ICR mice, and measured the amount of fluorescence remaining in blood and several organs at various time intervals. In blood, the level of Cy5.5-labeled HGC was the highest at 15 min, then after 30 min it decreased rapidly and reached a plateau form 30 min to 28 days. In the tissue we confirmed the presence of nanoparticles at high levels in the order of kidney>liver>submandibular gland until 28 days after injection. However, we did not find the presence of the particles in the brain or testes. These results will provide basic information on HGC as a drug delivery agent.

Antibiotic Detection Kit (Combination I), a lateral flow immunoassay (LFIA) developed for the detection of antibiotic residues in milk, was utilized for the analysis of antibiotic residues in the muscle tissue of olive flounder. After 60-min treatment by dipping in water dosed with ampicillin (200-g/ton water), the residue depletion of ampicillin was investigated in 25 cultured olive flounder (Paralichthys olivaceus). Muscles of fish were sampled on the 1st, 2nd, 3rd, 4th and 5th day after drug treatment. The concentration of ampicillin in the muscle was determined by LFIA. The absorbance ratio of the sample to the control blank (Bs/Bo) was employed as an index to determine the muscle residues in olive flounder. To investigate the recovery rate, standard solutions were added to muscle samples to give final concentrations in the muscle of 4 and 8 ng/ml. The recovery rates of all spiked samples were > 96% of the spiked value. Ampicillin was detected in the muscle of fish treated with the drug until the 2nd day of the withdrawal period. The present study showed that the LFIA can be easily adopted to predict ampicillin residues in tissue of farmed fishes.

Matrix metalloproteinases (MMPs) have been known to affect to cell migration, proliferation, morphogenesis and apoptosis by degrading the extracellular matrix. In the previous studies, undifferentiated mouse embryonic stem cells (ESCs) were successfully proliferated inside the extracellular matrix (ECM) analog-conjugated three-dimensional (3D) poly ethylene glycol (PEG)-based hydrogel. However, there is no report about MMP secretion in ESCs, which makes it difficult to understand and explain how ESCs enlarge space and proliferate inside 3D PEG-based hydrogel constructed by crosslinkers containing MMP-specific cleavage peptide sequence. Therefore, we investigated what types of MMPs are released from undifferentiated ESCs and how extracellular signals derived from various niche conditions affect MMP expression of ESCs at the transcriptional level. Results showed that undifferentiated ESCs expressed specifically MMP2 and MMP3 mRNAs. Transcriptional up-regulation of MMP2 was caused by the 3D scaffold, and activation of integrin inside the 3D scaffold upregulated MMP2 mRNAs synergistically. Moreover, mouse embryonic fibroblasts (MEFs) on 2D matrix and 3D scaffold induced upregulation of MMP3 mRNAs, and activation of integrins through conjugation of extracellular matrix (ECM) analogs with 3D scaffold upregulated MMP3 mRNAs synergistically. These results suggest that successful proliferation of ESCs inside the 3D PEG-based hydrogel may be caused by increase of MMP2 and MMP3 expression resulting from 3D scaffold itself as well as activation of integrins inside the 3D PEG-based scaffold.

본 연구에서는 심지층 처분장 부지선정 시에 고려되는 요소를 지질, 수리지질, 지화학 등으로 분류하고 그 첫 번째 단계로 지질분야의 세부 항목을 지형, 토양층, 암종, 구조지질, 역학적 안정성, 지질학적사건으로 분류하였으며, 이들 항목에 대한 국외 기준분석을 수행하였다. 부지선정요소(Siting factor)에 대한 기준(Criteria)은 각 국가의 처한 지질환경에 따라 다른 조건혹은 값을 제시하고 있다. 화산 및 지진활동이 빈번한 일본에서는 이에 대한 기준을 상대적으로 자세히 기술하고 있으며, 빙하작용이 예상되는 스웨덴에서는 빙하작용에 의한 지반 융기·침식에 대한 영향을 상세히 분석하였다. 따라서, 본 논문 결과는 향후 국내의 심지층 처분장 부지선정 기준 수립시에 중요한 참고자료로 활용될 수 있을 것으로 판단된다.

The present study purposed to examine the effects of transcutaneous electrical nerve stimulation, self-stretching and functional massage on the recovery of muscle contraction force for muscle fatigue caused by sustained isotonic contraction. The subjects of this study were 45 healthy students. They were divided into transcutaneous electrical nerve stimulation group(n=15), self-stretching group(n=15) and functional massage group(n=15), and using Primus RS. We observed the pattern of changes in maximal voluntary isometric contraction force(MVIC) after causing muscle fatigue in quadriceps femoris muscle through sustained isotonic contraction. Maximal voluntary isometric contraction force(MVIC) were greatly increased after transcutaneous electrical nerve stimulation, self-stretching and functional massage. In the comparison of recovery rate of muscle contraction force for muscle fatigue caused by sustained isotonic contraction among the treatment groups, it did not show any significant differences. However, it showed that each treatment may be effective in recovery of muscle fatigue caused by sustained isotonic contraction.

Semen can be divided into two parts. One is cellular part which contains sperms the other is liquid part which is called by seminal plasma. The seminal plasma is a nutritive and protective medium for the sperms. Fructose, which is major energy source, is supplied to sperms swim to female oocyte. Alkalic property protects sperms from hostile environment of female reproductive organ. Also, seminal plasma induces tolerance to preexisted immune cells, and changes intra‐uterine environment to better conditions for fertilized embryos to implant. However, the effects of seminal plasma in in vitro culture of fertilized embryos are unclear. Second fraction of fresh semen was obtained from a normal farm pig. The semen was centrifuged to remove sperms, and then supernatant was filtrated. The filtered seminal plasma was stored in — 30℃. In this study, electrically activated and chemically activated porcine embryos were employed to investigate the developmental rate after 2 hours treatment of none, 0.1%, 0.5%, and 1% seminal plasma in culture media by two days of activation. Both electrically and chemically activated embryos, cleavage rate and cell numbers of blastocysts were not significant difference within four groups. Blastocyst formation rate of electrically activated embryos also did not show significant difference within any groups. However 0.1% seminal plasma treatment group showed significantly increase of blastocyst formation rate in chemically activated group (None; 24.8%, 0.1%; 31.7%, 0.5%; 19.4, and 1%; 16.5%, respectively. p<0.05).

In the present study, we investigated the effect of porcine follicular fluid (PFF) concentration (10% vs. 1%) and protein-free media (PFF 0%) on maturation of porcine oocytes in vitro and analysed difference in gene expression in resulting blastocysts following parthenogenetic activation. Three groups were tested; 1) 10% PFF: Tissue culture medium (TCM) 199+10% PFF; 2) 1% PFF: TCM 199+1% PFF; and 3) 0.1% PVA: TCM 199+0.1 PVA. Cumulus-oocyte-complexes were cultured in the respective media containing gonadotrophin (1 ug/ml), epidermal growth factor (10 ng/ml), cystein (0.57 mM), sodium pyruvate (0.91 mM), insulin (5 ug/ml), 9-cis retinoic acid (5 nM) for 20～22 h and then without hormonal supplements for an additional 20-22 h. Data was analyzed using statistical analysis system(SAS) program. There was no significant difference in oocyte maturation rate. However, significantly higher (p<0.05) proportions of embryos developed to the blastocyst stage when oocytes were matured in 10% PFF group (45%) than in the 1% PFF group (31.1%). The total cell numbers were not significantly different among groups (52 ± 1.3 vs. 54.6±3.1 vs. 54.4±2.5, respectively). The relative abundance (ratio to beta-actin mRNA) of gene transcripts related to apoptosis in blastocysts was measured by real- time PCR. The expression of anti-apoptotic gene (BclxL) was up-regulated and the expression of pro-apoptotic gene (Bax) was down-regulated in 10% PFF group than in the other groups. Therefore, it can be concluded that supplementation of 10% PFF during in vitro maturation improves embryo development by reduction of apoptosis. * This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), MKE (#10033839-2011-13), Institute for Veterinary Science, the BK21 program and TS Corporation.

Embryo transfer (ET) is the final procedure for getting pregnancy through assisted reproductive technology such as IVF (in vitro fertilization), SCNT (somatic cell nuclear transfer). In our laboratory, the porcine cloned embryos loaded in ET medium are carried for 3 hours by portable incubator because of the great distance from the laboratory to the experimental farm. Thus, before transferring into recipient, porcine cloned embryos are exposed in vitro condition for long time. Medium which is used in this process is the TALP (Tyrode’s medium supplemented with 10 mM HEPES), but it includes little nutrients for embryo. Thus, the aim of this study is to determine whether ET media containing nutrients affect the in vitro development of embryos compared to TALP. For the experiment, porcine zygote medium (PZM)-5 which has amino acids for developing embryo was chosen as ET medium containing nutrients, added 10 mM Hepes as PZM-5 does not contain buffering system. For experiment, we carried out parthenogenesis through a chemical method using Thi/DTT. Parthenogenetic embryos were cultured in PZM-5 for 2 days, and then they were randomly divided into two group; loaded in a straw with TALP or PZM-5-Hepes, respectively. They were stored in a portable incubator for 3 hours to simulate the time consumed in ET, thereafter embryos in both TALP and PZM-5-Hepes groups were respectively cultured in PZM-5 for additional 5 days. All experiments were repeated 5 times. In result, blastocyst formation rate were 22.46%±1.47 and 23.17%± 2.13, respectively and total cell number were 32.9±2.22 and 37.09±2.18, respectively. There is no significant difference between TALP and PZM-5-Hepes groups. * Further study will investigate effect of PZM-5-Hepes on in vivo development of porcine cloned embryo. This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), Institute for Veterinary Science, the BK21 program and TS Corporation.

Adipose tissue-derived mesenchymal stem cells (ASCs) are very interesting in several laboratory animals and humans because they are easy to harvest and expand to generate millions of cells from a small quantity of fat. ASCs are known as useful materials for clinical applications in human cell therapy and as a donor cell in somatic cell nuclear transfer (SCNT). Here, we investigated if 1) minipig ASCs can be isolated, self-renewed and differentiated into multiple tissue lineages, 2) ASCs can be a suitable donor cell type for generation of cloned pig. In order to isolate ASC, adipose tissues were collected from inguinal region of a 6-year-old female minipig. The ASCs were attached to the culture dish with a fibroblast-like morphology. They expressed cell-surface marker characteristics of stem cell, underwent osteogenic, adipogenic, myogenic, neurogenic and chondrogenic differentiation when exposed to specific differentiation-inducing conditions. To investigate its potential as donor cell for cloning, we respectively carried out SCNT using ASC, adult skin fibroblast (ASF) and fetal fibroblast (FF) derived from same minipig. The ratio of blastocysts to 2-cell embryos and total cell number of blastocysts were monitored as experimental parameters. In results, cleavage and developmental competence to blastocysts rate showed no significant difference among the three groups. On the other hand, total cell numbers of blastocysts derived from ASC and FF were significantly higher than in ASF (89±7.9 and 105±5.5 vs. 57.5±5.2, respectively). Our results demonstrated that ASC have potential compared to ASF and FF in terms of the in vitro development and blastocyst formation ability. In further study, we will investigate the in vivo developmental ability of ASC as donor cell for pig cloning. * This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), Institute for Veterinary Science, the BK21 program, TS Corporation and Optifarm Solution.