Most traditional genome sequencing projects involving infectious viruses include culturing and purification of the virus. This can present difficulties as an analysis of multiple populations from multiple locations may be required to acquire sufficient amount of high-quality DNA for sequence analysis. The electrophoretic method provides a strategy whereby the genomic DNA sequences of the Korean isolate of Pieris rapae granulovirus (PiraGV-K) were analyzed by purifying it from host DNA by pulsed-field gel electrophoresis, thus simplifying sampling and labor time. The genomic DNA of infected P. rapae was embedded in agarose plugs, digested with a restriction nuclease and methylase, and pulsed-field gel electrophoresis (PFGE) was used to separate PiraGV-K DNA from the DNA of P. rapae, followed by mapping of fosmid clones of the separated viral DNA. The double-stranded circular genome of PiraGV-K encodes 120 open reading frames (ORFs), covering 92% of the sequenced genome. BLAST and ORF arrangement showed the presence of 78 homologs to other genes in the database. The mean overall amino acid identity of PiraGV-K ORFs was highest with the Chinese isolate of PiraGV (~99%), followed up with Choristoneura occidentalis ORFs at 58%. PiraGV-K ORFs were grouped, according to function, into 10 genes involved in transcription, 11 involved in replication, 25 structural protein genes, and 15 auxiliary genes. Genes for Chitinase (ORF 10) and cathepsin (ORF11), involved in the liquefaction of the host, were found in the genome. The recovery of PiraGV-K DNA genome by pulse-field electrophoretic separation from host genomic DNA had several advantages, compared with its isolation from particles harvested as virions or inclusions from the P. rapae host. We have sequenced and analyzed the 108,658 bp PiraGV-K genome purified by the pulsed field electrophoretic method. The method appears to be applicable to the analysis of genomes of large viruses. The chitinase, identified by PiraGV-K genome sequence, was functionally characterized by quantitative PCR, Western blot analysis, immunohistochemistry and transmission electron microscopy.

The cultivation methods of Acanthopanax in Korea need to be optimized. Hence, this study investigated the effect of different fertilizer ratios and planting dates on the growth and acanthoside D content of two (2) Acanthopanax species. The current recommended fertilizer rate of 10.5-8.5-8.5 kg/ha- (N-P2O5-K2O, respectively) produced the best plant growth of Acanthopanax. For the first year, the acanthoside D content resulting from the 2P (2x phosphate) rate was higher than that from the other fertilizer ratios, yet there were no significant differences resulting from the various treatments for either Acanthopanax divaricatus or Acanthopanax koreanum. Similarly, for the second year, there were no significant differences in the acanthoside D content resulting from the various fertilizer ratios, although for both species the acanthoside D content resulting from the 2P rate was slightly higher than that from the other treatments. Therefore, the results indicated that doubling the amount of phosphate increased the acanthoside D content. Plus, the optimum planting date with respect to growth and productivity for Acanthopanax divaricatus was identified as April 15. The cultivation methods of Acanthopanax in Korea need to be optimized. Hence, this study investigated the effect of different fertilizer ratios and planting dates on the growth and acanthoside D content of two (2) Acanthopanax species. The current recommended fertilizer rate of 10.5-8.5-8.5 kg/ha- (N-P2O5-K2O, respectively) produced the best plant growth of Acanthopanax. For the first year, the acanthoside D content resulting from the 2P (2x phosphate) rate was higher than that from the other fertilizer ratios, yet there were no significant differences resulting from the various treatments for either Acanthopanax divaricatus or Acanthopanax koreanum. Similarly, for the second year, there were no significant differences in the acanthoside D content resulting from the various fertilizer ratios, although for both species the acanthoside D content resulting from the 2P rate was slightly higher than that from the other treatments. Therefore, the results indicated that doubling the amount of phosphate increased the acanthoside D content. Plus, the optimum planting date with respect to growth and productivity for Acanthopanax divaricatus was identified as April 15. The cultivation methods of Acanthopanax in Korea need to be optimized. Hence, this study investigated the effect of different fertilizer ratios and planting dates on the growth and acanthoside D content of two (2) Acanthopanax species. The current recommended fertilizer rate of 10.5-8.5-8.5 kg/ha- (N-P2O5-K2O, respectively) produced the best plant growth of Acanthopanax. For the first year, the acanthoside D content resulting from the 2P (2x phosphate) rate was higher than that from the other fertilizer ratios, yet there were no significant differences resulting from the various treatments for either Acanthopanax divaricatus or Acanthopanax koreanum. Similarly, for the second year, there were no significant differences in the acanthoside D content resulting from the various fertilizer ratios, although for both species the acanthoside D content resulting from the 2P rate was slightly higher than that from the other treatments. Therefore, the results indicated that doubling the amount of phosphate increased the acanthoside D content. Plus, the optimum planting date with respect to growth and productivity for Acanthopanax divaricatus was identified as April 15. The cultivation methods of Acanthopanax in Korea need to be optimized. Hence, this study investigated the effect of different fertilizer ratios and planting dates on the growth and acanthoside D content of two (2) Acanthopanax species. The current recommended fertilizer rate of 10.5-8.5-8.5 kg/ha- (N-P2O5-K2O, respectively) produced the best plant growth of Acanthopanax. For the first year, the acanthoside D content resulting from the 2P (2x phosphate) rate was higher than that from the other fertilizer ratios, yet there were no significant differences resulting from the various treatments for either Acanthopanax divaricatus or Acanthopanax koreanum. Similarly, for the second year, there were no significant differences in the acanthoside D content resulting from the various fertilizer ratios, although for both species the acanthoside D content resulting from the 2P rate was slightly higher than that from the other treatments. Therefore, the results indicated that doubling the amount of phosphate increased the acanthoside D content. Plus, the optimum planting date with respect to growth and productivity for Acanthopanax divaricatus was identified as April 15. The cultivation methods of Acanthopanax in Korea need to be optimized. Hence, this study investigated the effect of different fertilizer ratios and planting dates on the growth and acanthoside D content of two (2) Acanthopanax species. The current recommended fertilizer rate of 10.5-8.5-8.5 kg/ha- (N-P2O5-K2O, respectively) produced the best plant growth of Acanthopanax. For the first year, the acanthoside D content resulting from the 2P (2x phosphate) rate was higher than that from the other fertilizer ratios, yet there were no significant differences resulting from the various treatments for either Acanthopanax divaricatus or Acanthopanax koreanum. Similarly, for the second year, there were no significant differences in the acanthoside D content resulting from the various fertilizer ratios, although for both species the acanthoside D content resulting from the 2P rate was slightly higher than that from the other treatments. Therefore, the results indicated that doubling the amount of phosphate increased the acanthoside D content. Plus, the optimum planting date with respect to growth and productivity for Acanthopanax divaricatus was identified as April 15.

The purpose of this study is to analysis of muscle fatigue in the upper trapezius and splenius capitis muscles according to therapy table height variation. The subjects were consisted of 15 healthy adults(10 males, 5 females) who had no medical history of neurological and musculoskeletal problems. In experiment, wireless electrode EMG system was measured for each the upper trapezius and splenius capitis muscles during the treatment performed on table. the differences in the muscle fatigue was compared for 4 types of table height(-6cm, -3cm, 0, +3cm from elbow in 90° flexion position). Muscle fatigue according to therapy table height were significant difference except for left upper trapezius. And muscle fatigue of right upper trapezius and splenius capitis showed significant decrease in +3cm table height compared to -6cm table height(p<.05). Muscle fatigue of right upper trapezius and splenius capitis were the highest in -6cm table height, but those were the lowest in +3cm table height. This study propose to change therapy table height higher than +3cm from elbow in 90° flexion position, if you hope to reduce muscle fatigue.

Emergence of resistant two-spotted spider mite (TSSM) can induce the over usage of standard amount of acaricides and result in various side effects. Rapid resistance monitoring is essential step for the efficient management of resistant populations by enabling the selection of appropriate acaricides. Here, we evaluated the 10 acaricides to determine its suitability on residual contact vial bioassay (RCV) by using PyriF strain as standard. Five acaricides (Abamectin, monocrotophos, tebufenpyrad, bifenazate and cyflumetofen) revealed the dose-dependent mortality within 8 h, whereas other remaining acaricides (Fenbutatin oxide, fenpyroxymate, flufenoxuron, spiromesifen and etoxazole) did not. This finding suggests that the application of RCV method is limited depending on the mode of action and physicochemical properties of each acaricide. Resistance levels to five acaricides (Abamectin, monocrotophos, tebufenpyrad, bifenazate and cyflumetofen) were determined for four field populations of TSSM by using RCV diagnostic kit. All TSSM populations showed the highest sensitivity to cyflumetofen, indicating that it would be most effective in controling field populations. RCV diagnostic kit would enable to provide crucial information for choosing the most appropriate acaricides in the field.

Currently, honeybee colonies are not stable and suffer from the infection of pathogens, affecting the pollination. For the alternatives to this difficulty, Bombus terrestris has been imported and used for pollination in agricultural fields. Although imported insects for pollination are very useful, the potential risk exposing to novel pathogens has been raised. To assess the risk primarily, we designed and synthesized PCR primers for detection of pathogens and parasites in B. terrestris. The samples were obtained from companies importing B. terrestris or field collections and genomic DNAs not showing physical shearing were purified. PCR for detection of pathogen- or parasite-specific gene revealed several DNA fragments were amplified in expected molecular size including Kashmir Bee Virus, Varroa jacobsoni, V. rindereri, Acarapis woodi and Aspergillus flavus. These amplified DNA fragments are in the process of cloning for DNA sequencing to confirm the target gene amplification. We also have plans to optimize the PCR conditions for each amplified target gene and try to develop biomarkers for diagnosis.

Environmental problem due to environmental pollution and CO2 emission has brought a strong attention on issues of sustainable urban development in urban planning area. Many urban planning researches have been argued that 'decentralized concentration' is one of the key issues for the sustainable urban spatial structure. Decentralized city is to create sustainable urban structure by realizing the supply system for urban energy efficiency through restriction of car use, proper urban density, and mixed-use. This study aims at exploring the concept of 'decentralized concentration' and comparing its characteristics of 'decentralized concentration' urban structure found in urban planning theories.

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.

The purpose of this study effectiveness of core strengthening exercise programs on symmetric double limb support and balance ability for elderly. The subjects that 30 persons between the ages of 65~80 elderly participated were divided into two groups randomly for 8 weeks. Tetrax interactive balance system and Berg's balance scale were used to assess support and stability. Paired t-tests were used to evaluate the changes before and after intervention. The difference between the groups was compared using an independent t-test. The experimental group showed significantly increase weight support, stability, balance(p<.05). However, the control group not showed significantly increase weight support, stability, balance(p>.05). In a variation, experimental and control groups showed significantly increased rate of weight support, stability, balance(p<.05). Consequently, core strengthening exercise program should be considered as a therapeutic method for the elderly to improve the balance ability and effectiveness on falls.

The faulty regulation of imprinting gene lead to the abnormal development of reconstructed embryo after nuclear transfer. However, the correlation between the imprinting status of donor cell and preimplantation stage of embryo development is not yet clear. In this study, to determine this correlation, we used the porcine spermatogonial stem cell (pSSC) and fetal fibroblast (pFF) as donor cells. As the results, the isolated cells with laminin matrix selection strongly expressed the GFRα-1 and PLZF genes of SSCs specific markers. The pSSCs were maintained to 12 passages and positive for the pluripotent marker including OCT4, SSEA1 and NANOG. The methylation analysis of H19 DMR of pSSCs revealed that the zinc finger protein binding sites CTCF3 of H19 DMRs displayed an androgenic imprinting pattern (92.7%). Also, to investigate the reprogramming potential of pSSCs as donor cell, we compared the development rate and methylation status of H19 gene between the reconstructed embryos from pFF and pSSC. This result showed no significant differences of the development rate between the pFFs (11.2±0.8%) and SSCs (13.3±1.1%). However, interestingly, while the CTCF3 methylation status of pFF-NT blastocyst was decreased (36.3%), and the CTCF3 methylation status of pSSC-NT blastocyst was maintained. Therefore, this result suggested that the genomic imprinting status of pSSCs is more effective than that of normal somatic cells for the normal development because the maintenance of imprinting pattern is very important in early embryo stage.

This study aims to reveal how EA affects BAX and NF-kB involved in cell deaths from global ischemia, and to do this, observes the changes of BAX and NF-kB caused by EA application after transient global ischemia. The experimental method is to give rise to global ischemia and apply EA to 27 SD rats with the particulars of being six-week-old, male, around-300 gram-weighing, and adapted to laboratory environment for more than a week, and divide them into three groups, that is, GV20 EA group(n=9), L14 EA group(n=9), no-treatment GI group(n=9), and then observe their changes of BAX and NF-kB at the time lapse of 6 hours, 9 hours and 12 hours after ischemia, using western blotting. The numerical decrease of BAX expression at the time lapse of 9 hours after EA application, though not statistically significant, was observed in GV20 EA group and L14 EA group, and the NF-kB expression appeared statistically significant decrease in GV20 EA group and L14 EA group, but the expression was higher in the group with EA application. Therefore, EA application at the early phase of global ischemia is considered to affect BAX and NF-kB and play a positive role in decreasing apoptosis and cell deaths by inflammation.

Insects or insect remains found in beer are one of major issues in consumer claim. Accurate estimation of inflow time isa critical factor for the settlement of such claims related with beer-contaminating insects but no reliable methods have been developed. In an attempt to establish a molecular marker-based diagnostic method, the degradation rates of 18S rRNA genes in the insectssoaked in 500 ml beer were investigated by quantitative real-time PCR (qPCR) over one month period at room temperature. Among the six insect species tested, the house fly (Musca domestica) and honey bee (Apis mellifera) revealed high correlations (r2=0.974-0.990) between the degradation of 18S rRNA gene and inflow time. In these insects, statistically significant distinction was possible between the samples stored in beer less than 14 days and more than 14 days. Other insects, including the fruit fly, common house mosquito, German cockroach and Indian meal moth, displayed poor correlations, which appeared attributed to the inefficient genomic DNA extraction likely due to small sample size or disintegration of body parts during storage in beer. With proper improvement in DNA extraction, this 18S rRNA-based diagnostic method would be applicable for estimating the inflow time of beer-contaminating insects.

The purpose of this study was to determine the effects of heat appli˗ cation on the immune activities of the human body. To exam, further˗ more, the immune effect from the healthy volunteer(male:15, female:15) by monitoring changes of immune substances such as various leukocytes[total white blood cell(WBC), eosinophil, neutrophil, basophil, monocyte, and lymphocyte], a comparative study with warm water immersion(40.8±0.3℃) and infrared(250W) was carried out. The plasma analysis showed that the count of white blood cell, eosinophil, and neutrophil were elevated in warm water immersion- or infrared˗ stimulated group compared with control group. However, the count of basophil was decreased in both warm water immersion- and infrared-stimulated group than control group. Therefore, these results suggest that the thermostimulation improved immune activity.

The Mental Image is designed and proposed for making a comfort and amenity in built environments using digital color analysis in the regional urban artificial environment. For a long time, it is crucial to relate visual environment with thermal environments. Especially, color makes an important role to making thermal comfort. In order to making this information, this paper describes how we can analyze the color information of the regional urban artificial environment using the color syntax program and transfer the data to form maker and create the mental image. Because of the rapid urbanization and the acceptance of mixed function, modern urban landscape is disordered and is not controlled. The Mental Image prepares our environmental color system corresponding to our mental image with perceiving regional urban environment color. This research aims to systematize the process making the Mental Image in the regional urban landscape’s color and suggest a theoretical basis to extract, embody and use the information of the regional main color from urban landscape color. Through these concepts, this research will suggest how to systematize, develop the Mental Image with emotional environmental color system. Finally, it is expected to offer a theoretical basis to embody the Mental Image and build up the emotional environments.

This study was conducted to examine the effect of IRES controlled reporter gene on screening and production of recombinant human erythropoietin (EPO) proteins from cultured CHO cells. The cDNA was cloned for EPO from human liver cDNA. Using site-directed mutagenesis, we generated recombinant human EPO (rhEPO) with two additional N-glycosylations (Novel erythropoiesis-stimulating protein: NESP). Wild type hEPO and NESP were cloned into expression vectors with GFP reporter gene under regulatory control of CMV promoter and IRES so that the vectors could express both rhEPO and GFP. The expression vectors were transfected to cultured CHO-K1 cells. Under microscopy, expression of GFP was visible. Using supernatant of the culture, ELISA assay, immunocytochemistry and in vitro assay using EPO dependant cell line were performed to estimate biological activity to compare the production characteristics (secretion levels, etc.) between rhEPO and NESP. The activity of NESP protein, obtained by mutagenesis, was described and compared with its rhEPO counterpart produced under same conditions. Although NESP had less secretion level in CHO cell line, the biological activity of NESP was greater than that of rhEPO. These results are consistent with previous researches. We also demonstrated that rhEPO and GFP proteins expressed simultaneously from transfected CHO cell line. Therefore we conclude that use of GFP reporter gene under IRES control could be used to screen and produce rhEPO in cultured CHO cells.

고무화합물 형태로 구성된 조영제의 병에 Syringe Connector의 Spike를 연결 시 고무의 찢김 정도를 알아보고 찢김 및 분쇄로 인한 합성고무의 혼입 유무와 분쇄된 합성고무가 검출 시 분쇄물의 크기를 실험을 통해 알아보고자 하였다. 그 결과 찢김 정도의 경우 Syringe Connector의 끝과 최초 접촉하는 앞면이 약 3.14±0.04 ㎜로 뒷면 보다 많이 찢겼으며, 실험 대상인 10 병의 조영제에서 평균 7 개에서 15 개로 모두 분쇄물이 검출되었다. 검출된 분쇄물을 이용하여 크기를 측정한 결과 평균크기는 약 7.89±0.31 ㎛이었다. 향 후 다양한 실험 및 분석방법을 통한 추가실험과 더불어 흡인된 분쇄물 차단을 위한 미세 필터타입 자동주입장치의 개발이 필요하며, 분쇄물 유입 시 치명적 사고를 대비하여 관련기관의 관심 또한 필요할 것으로 사료된다.