Setdb1/Eset, a histone lysine methyltransferase, is recruited by various transcription factors to modify local chromatin. The observation that Setdb1-null blastocysts fail to produce epiblast-lineage cells suggests a role for Setdb1 in generating mouse embryonic stem cells (mESCs). When examined in mouse zygotes, Setdb1 proteins appeared as dots at perinucleolar rims of pronuclei, with the dot-shaped signals more prominent in male pronuclei. Setdb1 signals were observed diffusely in the nucleus from the two-cell stage onward and, by the blastocyst, took a punctate form, away from nucleolus. Such varying expression patterns suggest its involvement in diverse biological processes at preimplantation stage. Setdb1 appeared in Oct4-positive cells of inner-cell-mass origin but not in trophectoderm-lineage cells in blastocyst outgrowths. Setdb1 co- immunoprecipitated with Oct4 in mESCs, and Setdb1 expression was markedly reduced upon retinoic acid- induced differentiation. These observations suggest that Setdb1 has an important role in maintaining the self-renewal of mESCs through collaboration with Oct4.

Microtubule-associated protein 1B (MAP1B), a member of MAP1 family, plays a key role in the brain development. MAP1B binds to many kinds of proteins directly or indirectly. In our previous studies, MAP1B and glyceraldehydes 3-phosphate dehydrogenase (GAPDH) were down-regulated in bovine follicular cystic follicles (FCF). This study was performed to examine interaction between MAP1B and GAPDH in bovine follicles using immunoprecipitation (IP) with western blot analysis and immunohistochemisty. MAP1B and GAPDH mRNA expression levels were down-regulated in bovine FCFs. Consistent with the semi-quantitative PCR data, their protein expressions were also down-regulated in FCFs. IP data showed that MAP1B bound to GAPDH in normal follicles, but their binding was absent in FCFs, suggesting that these data might be resulted from a low level of MAP1B and/or GAPDH expression. These results suggest that GAPDH does not as always function as a loading control in bovine follicles.

Sweet potato whitefly (SPW), Bemisia tabaci, is the most serious pest of greenhouse tomato as a vector of tomato yellow leaf curl virus since 2008 in Jeju. This study was conducted for within-plant distribution of SPW nymphs on leaves and the characteristic of spatial distribution of SPW adults using by yellow-color sticky trap in greenhouse-grown tomato plants. The nymphs of SPW were mainly distributed on leaves of mid-lower stratum in three stratum of tomato plant. They were also more distributed on three to four leaflets from basal part in a leaf. Ten plots and three plants per plot were appropriated to estimate of mean density of SPW nymph using by visual inspection in a tomato greenhouse. The more SPW adults were caught on horizontally placed sticky trap above 50～80 cm than vertically placed sticky trap above tomato canopy. The horizontal direction trap should be placed in opened site. SPW adults were shown the aggregated distribution pattern using by Taylor’s power law (TPL) and Iwao’s patchiness regression (IPR) model which are commonly used for spatial distribution pattern analysis. TPL was showed better description than IPR for spatial distribution pattern of SPW adults. The required number of trap tended to decrease according to increase the mean density of SPW adult

Gene targeting is a genetic technique that utilizes homologous recombination between an engineered exogenous DNA fragments with the endogenous genome of an organism. In domestic animal, gene targeting has provided an important tool for producing Knock-out pig for GGTA1 gene to use xenotransplantation. The frequency of homologous recombination is a critical parameter for the success of gene targeting. The efficiency of homologous recombination in somatic cells is lower than that in mouse ES cells. So the application of gene targeting in somatic cells has been limited by its low efficiency. Recently, knock-out rat and mouse was generated by introducing nonhomologous end joining (NHE)-mediated deletion or insertion at the target site using zinc-finger nucleases (ZFN). Therefore, the development of effective knock-out and knock-in techniques in domestic animal is very important in biomedical research. In this present study, we investigated whether homologous recombination events occurs at cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) gene locus using ZFN in porcine primary fibroblast. CMAH-targeted ZFN DNA and mRNA were purchased from SIGMA-Aldrich. CMAH neo targeting vector consists of the neomycin resistance gene as a positive selectable marker gene, 789 bp 5’ arm and 763 bp 3’ arm from Exon 8 of CMAH gene. For transfection, the targeting vector and ZFN DNA or mRNA were introduced into ear fibroblasts cells of Chicago miniature pig by electroporation. After selection of G-418, PCR analysis was performed using 213 colonies transfected with ZFN DNA or mRNA. As a result, 39 positive colonies were identified in colonies transfected with ZFN DNA or mRNA. To our knowledge, this study provides the first evidence that the efficiency of gene targeting using ZFN was higher than that of conventional gene targeting in the porcine fibroblast. These cell lines may be used in the production of CMAH knock-out for xenotransplantation.

Entomopathogenic fungus Aschersonia aleyrodis naturally occurred on citrus whitefly, Dialeurodes citri nymph was often observed in organic citrus orchards, Jeju. The genus Aschersonia is also known to be toxic against scale insects and other pests. However, little is known about artificial media for mass production of spores of Aschersonia species. Grains are excellent sources of media for mass conidia production of various entomopathogenic fungi. The yeast extract, which converts carbohydrates to carbon dioxides and alcohols, contains a large amount of vitamin B complexes which facilitate the carbohydrate metabolism. The more yeast extract content the more conidia production on artificial medium made from commercial corn flour and corn gluten feed. The number of conidium produced on oat, millet, sorghum, and unhulled barley medium containing 1% yeast extract were 1.8, 1.8, 1.6, and 2.1×1010/plate (90mm × 15mm), respectively. However, the greatest yeast effect among four media showed appeared on sorghum medium, which produced 25 times higher spore production than sorghum alone. Furthermore, the conidia from solid sorghum medium could be easily harvested with cell scraper.

Aschersonia aleyrodis was well-known to be a biological control agent for citrus whitefly, Dialeurodes citri. This entomopathogenic fungus is naturally occurred in organic farming citrus orchards in Jeju. Both lime-sulfur and Bordeaux mixtures are extensively used today to control citrus diseases like citrus melanose and citrus scab, especially in organic farming pest management program. The high concentrated lime-sulfur is also used for pest control such as pink citrus rust mite and scale insects. This study was focused to test the conidial germination and sporulation of Aschersonia aleyrodis on potato dextrose agar (PDA) medium containing different concentrations of two fungicides. The conidia of Aschersonia aleyrodis grown on PDA mixed with commercial bordeaux mixtures, CM150-505, at dilution rate 1:200 (water : bordeaux mixture) were well-germinated but not sporulated at all. On the other hand, Aschersonia aleyrodis did not sporulate and germinate on lime sulfur treated PDA medium even at extremely low dilution rate 1:2,048,000 (water : lime sulfur). However, when the mycelial cells grown at 25o C for at least 7 days at soluble starch-tryptone medium were added to PDA, they were well-sporulated even at high dilution rate 1:100 (water : lime sulfur). This result suggested that the spore mixtures of Aschersonia aleyrodis should be applied to field quite long after lime-sulfur spray.

Riptortus clavatus is one of the most serious pests affecting the cultivation of sweet persimmon, which is one of the most important export fruits in Korea. A pheromone-baited trap has been used in attempts to control this pest. Although the trap showed relatively good trap efficacy, it was often observed that R. clavatus walks around and rests on the surface of the trap rather than entering the trap. In the present study, the development of a more efficient pheromone-baited trap for R. clavatus was pursued by investigating 3 different traps: the fish trap, the pyramidal trap, and the symmetry trap. The fish trap captured the lowest number of R. clavatus, and the pyramidal trap showed moderate capture efficacy. Significantly greater numbers of R. clavatus adults were captured in the symmetry trap. Significantly increased trapping efficacy of the symmetry trap would be useful for mass trapping of R. clavatus.

Beet armyworm (BAW), Spodoptera exigua (Hübner), is a polyphagous lepidopteran pest mainly distributed in tropical and subtropical regions. BAW are known to migrate long distances to find suitable host plants and environmental condition. The possibility of its overwintering in temperate regions has been suspected. This study was conducted for investigating overwintering and phenology of BAW using by sex pheromone trap in Jeju during 2007 to 2011. BAW was able to overwinter at pupae stage regardless of temperature of winter season in Jeju. The times of BAW caught first on traps were different by surveyed years - early March in 2007, middle March in 2008 and 2011 and early April in 2009 and 2010. The times of BAW caught on trap tended to be more dependent on temperature variation during on November and December than on mean temperature of winter season. The peak season of adults of the overwintering generation was early to middle of May. The results of seasonal periodicity of BAW monitored by sex pheromone trap during 2009 and 2010 indicated that five generations were passed in Jeju. The peak season of 1st generation was on middle of April, 2nd generation on middle of July, 3rd generation on early of August, 4th generation on late of August and 5th generation on middle of September. There were similar patterns by different above sea levels except the season of overwintering generation adults occurrence (March to May).

Arrowhead scale, Unaspis yanonensis, is a serious scale insect pest of citrus in Jeju. The sticky taping of 1.0 cm diameter twig was a good survey method for arrowhead scale crawler. There are three times crawler occurrence season - late May to early July, late July to middle September and late September to late October. But the third occurred crawlers are all died during overwintering in open-field groves. This study was conducted to provide information on efficient sampling plan for arrowhead scale within the tree. Twig and fruit was fit for secondary sample unit by two-stage variance analysis. Both sample number per tree (primary sample unit) was eight twigs or fruits. Taylor’s power law was better described the distribution characteristic of arrowhead scale than Iwao’s patchiness regression. The slope of both linear regressions was greater than “1” which means aggregative distribution pattern. Minimum sample size to estimate the density was calculated using by two parameters of Taylor’s power law. To estimate the mean density on twig and fruit by binomial sampling plan, the optimum tally threshold was 5 and 7, respectively.

공학적 안전설비 공기정화계통과 관련된 실험 수행을 위해 원자력등급 ESF 공기정화계통 시뮬레이터 를 설계, 제작, 검증하였다. 영광 5,6호기 주제어실 공기정화계통의 공급자 정보, 도면 등을 기준으로 실 사를 통해 치수를 확인하여 3D CAD 모델을 작성하였다. 모델과 현장 계통의 실측 유량을 기준으로 CFD 분석을 수행하였다. 공기정화계통으로 유입되는 공기는 30 ℃, 유동형태는 균일한 것으로 가정하고, 검사 기록지에 의한 주제어실 ESF 공기정화계통의 유량이 12,986 CFM이고, 610×610 ㎟의 HEPA 필터가 9개 설치되어 있으므로 HEPA 필터 단면를 지나는 유속은 1.83 m/s이다. 주제어실 공기정화계통 모델링시 공 기 유동이 흐르지 않는 필터 테두리 지지대를 고려하여 현장과 유사한 유동현상을 모사하였다. 약 8 m/s 로 기록된 활성탄 흡착기 하단의 공기유동은 별도의 분석을 통해 7 m/s 이상의 유속이 모사되도록 CFD 분석하였다. 연료건물 비상배기계통 및 비상노심냉각계통 기기실 배기정화계통의 공기정화계통에 대해 서도 CFD 분석한 결과, 시뮬레이터의 유속을 조절하면 세가지 ESF 공기정화계통을 모두 모사할 수 있음 을 확인하였다. CFD 분석 후 시뮬레이터를 원자력등급으로 제작하였고, 본 실험에 착수하기 전에 공기유 동 분포도실험을 통해 시뮬레이터의 신뢰도를 검증하였다. 검증결과 중급 필터를 장착한 상태에서 시뮬 레이터의 필터 지지대 부분을 제외한 내부에서 공기유동이 고르게 분포함을 확인하였고, 제작된 시뮬레 이터는 Reg. Guide 1.52(Rev.3) 개정 내용 확인을 위한 실험에 사용되었다.

공학적 안전설비 공기정화계통의 규제지침인 Reg. Guide 1.52(Rev.3)의 변경사항중 성능시험과 관련 한 운전가능성 시험시간 단축, HEPA 필터 현장누설시험용 시험물질 변경 및 활성탄 성능시험 Methyl Iodidie 투과허용율 상향 변경을 영광 5,6호기에 적용하고자, 모사실험장치와 현장 설비를 활용하여 기술 적 타당성을 확인하는 실험을 수행하였다. 10시간 이상의 장시간 운전가능성 시험을 통해 계통내 습분을 제거하여도 시험후 1∼4일만에 회복됨을 확인하여 운전가능성 시험은 기기적 운전가능성 점검에 적합한 매월 15분 이상의 시험을 수행하는 것이 타당함을 확인하였다. HEPA 필터 현장누설시험용 시험물질 변 경을 위해 DOP와 PAO의 에어로졸 입자크기, 발생량, 누설인지도를 비교한 결과 PAO는 원전에서도 DOP 대체시험물질로 사용 가능함을 확인하였다. 베드깊이 4 인치 이상의 활성탄여과기에 대한 Methyl Iodide의 투과율 허용치가 0.175 %에서 0.5 %로 상향 변경된 것은, ASTM D3803(1989)으로의 활성탄 성 능시험 방법 변경에 따른 것으로서, 30 ℃ 상대습도 95 %에서의 Methyl Iodide 투과허용율 0.5 %가 사용 중 활성탄의 성능을 시험하기에 충분히 보수적인 시험방법임을 확인하였다. 본 실험 결과를 바탕으로 영 광 5,6호기는 인허가변경을 완료하였다.

Enterococcus faecalis, a gram-positive bacterium, has been implicated in endodontic infections, particularly in chronic apical periodontitis. Proinflammatory cytokines, including tumor necrosis factor-α (TNF-α), are involved in the pathogenesis of these apical lesions. E. faecalis has been reported to stimulate macrophages to produce TNF-α. The present study investigated the mechanisms involved in TNF-α production by a murine macrophage cell line, RAW 264.7 in response to exposure to E. faecalis. Both live and heat-killed E. faecalis induced high levels of gene expression and protein release of TNF-α. Treatment of RAW 264.7 cells with cytochalasin D, an inhibitor of endocytosis, prevented the mRNA up-regulation of TNF-α by E. faecalis. In addition, antioxidant treatment reduced TNF-α production to baseline levels. Inhibition of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein (MAP) kinase also significantly attenuated E. faecalis-induced TNF-α expression by RAW 264.7 cells. Furthermore, activation of NF-κB and AP-1 in RAW 264.7 cells was also stimulated by E. faecalis. These results suggest that the phagocytic uptake of bacteria is necessary for the induction of TNF-α in E. faecalis-stimulated macrophages, and that the underlying intracellular signaling pathways involve reactive oxygen species, ERK, p38 MAP kinase, NF-κB, and AP-1.

Effects of Vegemil® containing soybean proteins and isoflavones on the growth and bone density of broiler chickens were investigated. One-week-old male and female Arbor Acres broiler chickens were fed on Vegemil® A containing 3% soybean proteins and 162 ppm isoflavones, instead of water, for 30 days and their growth indices (body weight, leg weight and femur length) and bone density were analyzed. The body weight gains in male and female chickens were increased by 15.6% and 31.7%, respectively, following feeding Vegemil® A compared to normal water. Vegemil® A increased leg weight as well as femur length of females by 22.9% and 15.0%, respectively. In addition, Vegemil® A feeding enhanced femoral bone density by 21.3% in comparison with water feeding. Therefore, it is suggested that Vegemil® A not only facilitates body growth, but also strengthens bone density of normal chickens, and that it could be a promising candidate for the improvement of infant growth and for the prevention of menopausal osteoporosis.

Porphyromonas gingivalis is a major etiologic agent of chronic periodontitis and cytokines produced by macrophages play important roles in the pathogenesis of periodontal diseases. In this study we investigated the cytokine response of phorbol myristate acetatedifferentiated THP-1 cells exposed to P. gingivalis. Compared with the prominent cell wall components of P. gingivalis (lipopolysaccharide and the major fimbrial protein FimA), live P. gingivalis stimulated much higher levels of cytokine production. In addition, whereas low multiplicity of infection challenges (MOI=10) of P. gingivalis 381 stimulated high levels of monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β, high dose challenges with this bacterium (MOI = 100) resulted in a substantially diminished production of MCP-1 and IL-6. Moreover, high MOI P. gingivalis challenges achieved only low levels of induction of MCP-1 and IL-6 mRNA. The decreased production of MCP-1 and IL-6 appeared to be mediated by P. gingivalis proteases, because high MOI challenges with congenic protease mutant strains of this microorganism (MT10 and MT10W) did not result in a diminished production of MCP-1 and IL-6. Similar to its protease mutant strains, leupeptin (a protease inhibitor)- treated P. gingivalis at high doses induced high levels of MCP-1 production. To examine the mechanisms underlying the diminished production of MCP-1 by P. gingivalis proteases, the activation of mitogen-activated protein (MAP) kinases and NF-xB was compared between the 381 and MT10W strains. Whilst high doses of both 381 and MT10W similarly activated the three members of the MAP kinase family, the DNA binding activity of NF-xB, as revealed by gel shift assays, was greatly increased only by MT10W. Taken together, our data indicate that P. gingivalis stimulates the production of high levels of TNF-α, IL-1β, IL-6, and MCP-1 but that high dose challenges with this bacterium result in a diminished production of MCP-1 and IL-6 via the protease-mediated suppression of NF-B activation in THP-1 macrophagic cells.