In Korea, the construction of dry storage facilities for spent nuclear fuel is being promoted through the 2nd basic plan for high-level radioactive waste management. When operating dry storage facilities, exposure dose assessment for workers should be performed, and for this, exposure scenarios based on work procedures should be derived prior. However, the dry storage method has not yet been sufficiently established in Korea, so the work procedure has not been established. Therefore, research is needed to apply it domestically based on the analysis of spent nuclear fuel management methods in major overseas leading countries. In this study, the procedure for receiving and storing spent nuclear fuel in a concrete overpack-based storage facility was analyzed. Among the various spent nuclear fuel management systems, the metal overpack-based HI-STAR 100 system and the concrete overpackbased HI-STORM 100 system are quite common methods in the United States. Therefore, in this study, work procedures were analyzed based on each final safety analysis report. First, the HI-STAR 100 overpack enters the facility and is placed in the transfer area. Remove the impact limiter of the overpack and install the alignment device on the top of the overpack. Place the HI-TRAC, an on-site transfer device, on top of the alignment unit and remove the lids of the two devices to insert the canister into the HI-TRAC. When the canister transfer is complete, reseat the lid to seal it, and disconnect the HI-TRAC from the HI-STAR 100. Raise the canister-loaded HI-TRAC over the alignment device on the top of the HI-STORM 100 overpack and remove the lids of the two devices that are in contact. Insert the canister into the HI-STORM 100 and reseat the lid. The HI-STORM 100 loaded with spent nuclear fuel is transferred to the designated storage area. In this study, the procedure for receiving and storing spent nuclear fuel in a concrete overpack-based storage facility was analyzed. The main procedure was the transfer of canisters between overpacks, and it was confirmed that HI-TRAC was used in the work procedure. The results of this study can be used as basic data for evaluating the exposure dose of operating workers for the construction of dry storage facilities in Korea.

점착트랩은 특정 광파장에 유인되는 곤충의 반응을 이용하여 비행 곤충의 유살 및 모니터링에 널리 이용되고 있다. 그러나 유색 점착트랩은 해당 색상에 반응을 보이는 모든 곤충을 유인하기 때문에 비표적곤충(Non target insect)이 부착되는 문제가 발생한다. 비표적곤충은 부착면을 차지하여 표적곤충의 유인을 방해하며, 해충밀도 모니터링에 필요한 노력과 시간 소요를 증대시킨다. 본 연구에서는 비표적 곤충과 이물질의 부착을 물리적으로 방지하기 위하여 내경 5mm, 10mm의 방지망을 설치하였을 때, 각 처리에 따른 유입 곤충의 종다양성 지수를 분석하여 방지망의 현장 적용 가능성을 검정하였다. 그 결과 망을 처리하지 않은 트랩과 5mm, 10mm 방지망을 설치한 트랩 간 종다양성 지수에서 유의한 차이를 보이지 않았다. 그러나 포획된 총 종의 수는 망을 처리하지 않은 트랩에서 높게 나타났는데 이는 대형 비표적 곤충의 포획수가 높은 것이 주 요인이었다. 표적 해충인 총채벌레, 가루이 등 미소해충의 유입 빈도 또한 무처리, 5mm, 10mm 간에 유의한 차이를 보이지 않아 방지망의 설치가 표적곤충인 미소해충의 유입에 악영향이 없으며, 5mm 망을 적용하여도 무방하다는 결과를 얻었다.

Recent research on stem cell conditioned medium (CM) has been revealed that CM could influence on the embryo development when supplemented to in vitro culture medium. However, the optimal basal medium for CM production has not determined although it is the fundamental factor of CM. The purpose of this study is to examine the effect of human derived adipose stem cell CM (hASC-CM) with different basal medium on mice embryo development after parthenogenetic activation (PA). hASC-CM was collected from 2 kinds of serum free basal medium, DMEM and KSFM, respectively on day 5 from the culture of hASC isolated from human fat tissue. Intra-peritoneal injection of PMSG and hCG was conducted into 7-week-old ICR mice for superovulation. The oocytes were recovered from the oviductal ampulla, 18 h after hCG injection, and denuded using 0.1% hyaluronidase. PA of oocytes was conducted with KSOM media including strontium chloride. The parthenotes were in vitro cultured in 3 groups: 100% KSOM (Control), 75% KSOM + 25% DMEM or KSFM without FBS (DMEM or KSFM group) and 75% KSOM + 25% hASC-CM from DMEM or KSFM (DMEM-CM or KSFM-CM group). Cleavage rate was assessed after 2 days post IVC and blastocyst formation rate was evaluated after 6 days post IVC both using stereomicroscope. Total cell number of blastocysts was counted by Hoechst staining. 1way ANOVA from Graphpad prism 5 was used for statistical analysis and the values are presented as means ± standard error of mean. As a result, blastocyst formation rate of DMEM-CM group (16.09±3.32%, P<0.05) was significantly lower than control and DMEM group (34.43±2.89% and 34.49±5.34%, P<0.05) but cleavage rate and total cell number of blastocysts showed no significant difference among groups. In case of KSFM, there was no significant difference in cleavage rate, blastocyst formation rate and total cell number of blastocysts among the control, KSFM group and KSFM-CM group. The sort of basal medium used for the CM collection affected the development of parthenotes during in vitro culture differently. Therefore, further research should be conducted to find out the alternative basal medium of CM able to improve the embryo development. This research was supported by Nature Cell (#550-20170028), Cooperative Research Program of RDA (CCAR, #PJ013954022018), Research Institute for Veterinary Science and the BK21 plus program.

A recent study reported that Pleurotus ostreatus has the potential to be used as a β-glucan-based cream for supportive complementary therapy of atopic dermatitis. KH054 is a new herbal prescription consisting of P. ostreatus and Panax ginseng. The effects of atopic dermatitis-induced materials on the expression of cytokine genes in human monocytes (THP-1, EoL- 1) have been examined. Some reports demonstrated that P. ginseng augments the activity of natural killer cells, which plays an important role in innate immunity against infection and tumor development. Monocyte chemotactic protein 1 (MCP-1), interleukin (IL)-6, and IL-8 have important roles in mediating the infiltration of various cells into the skin of atopic dermatitis and psoriasis. The present study investigated whether KH054 on induced IL-6, IL-8, and MCP-1 secretion by house dust mite (Dermatophagoides pteronissinus) in THP-1 (human acute monocytic leukemia) and EoL-1(Human eosinophilic leukemia) cell. D. pteronissinus functions in the pathogenesis of allergic diseases, including atopic dermatitis and asthma. The inhibitory effect of KH054 on the induction of IL-6, IL-8, and MCP-1 secretion by D. pteronissinus extract in THP-1 and EoL-1 cells was examined. KH054 potently suppressed the elevated production of IL-6 and IL-8 induced by D. pteronissinus treatment in THP-1 and EoL-1 cells. Based on the present results, KH054 may be useful for developing functional foods to treat atopic dermatitis.

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.

This experiment was conducted to determine the effects of applying level of pig slurry on the characteristics of runoff and leaching water in lysimeter. Treatments were broken down into five treatments such as non-chemical fertilizer, chemical fertilizer (200㎏ N/㏊) and pig slurry (200, 400, 600㎏ N/㏊), and 3-time-repeated randomized complete block design was fulfilled. The concentrations of NO₃-N was raised(p＜0.05) in proportion to the applying level of pig slurry. NO₃-N concentration of leaching water collected from soil depth 20㎝ and 40㎝ was intensified as the applying level of pig slurry was higher (p＜0.05) in pig slurry 600㎏ N/㏊ application. In conclusion, pig slurry at the volcanic ash soil in Jeju area can replace the chemical fertilizer as it is tested that applying 200㎏ N/㏊ of pig slurry and chemical fertilizer show the same productivity of sorghum×sudangrass hybrid. But more than 200㎏ N/㏊ of pig slurry may not be appropriate because it may contaminate the runoff and leaching water even though it may increase for the forage productivity.

Background : Rehmannia glutinosa is a perennial herb belonging to the family Scrophulariaceae. Its roots have been utilized as a traditional medicine but the aerial parts (flower, flower stalk, leaf) were not used. In this paper, we aimed to determine the content of three compounds [aucubin, catalpol, and γ-aminobutyric acid (GABA)] in different organs of R. glutinosa. Methods and Results : The flower, flower stalk, leaf, and root of R. glutinosa were harvested in the end of August. The aucubin and catalpol were analyzed by LC/MS, whereas GABA was analyzed by GC/MS. The aucubin content was the highest in the leaf, while catalpol and GABA were the highest in the flower. The aucubin content of in the leaf was 1.43, 0.81, and 1.07 ㎎/g, respectively. The catalpol content of flower in Dakang, Tokang, and Suwon 9 was 41.06, 28.78 and 37.48 ㎎/g, respectively. The GABA content of flower in Dakang, Tokang, and Suwon 9 were 0.79, 0.76 and 0.65 ㎎/g, respectively. Conclusions : The contents of aucubin, catalpol, and GABA were higher in leaf and flower than that of root. This study provides the important information of R. glutinosa leaf and flower as a potential supplement.

Background : Ginseng has been commonly used as a traditional oriental medicine for its wide spectrum of medicinal properties, including anti-inflammatory, antitumorigenic, adaptogenic and anti-aging properties. 20(S)-Protopanaxadiol (PPD), a intestinal metabolite of ginsenosides, is one of the active ingredients in ginseng. In this study, we have found inflammation-related genes regulated by 20(S)-PPD in mouse bone marrow-derived macrophage (BMDM) to elucidate the role of 20(S)-PPD in inflammatory signaling pathways. Methods and Results : We examined cell viability of BMDM cells after treatment of 20(S)-PPD and found that 20(S)-PPD has no cytotoxicity up to 10 uM. BMDM cells treated with none or 10uM 20(S)-PPD were used for RNA extraction and microarray analysis. It was found that 2 inflammation-related genes are upregulated and 4 genes are downregulated by 20(S)-PPD. Conclusion : These results can give clues to elucidate the role of 20(S)-PPD in inflammatory signaling pathways.

Background : Ginseng, one of most famous traditional oriental medicines, has been known for a number of pharmacological properties including anti-tumor, anti-diabetic, anti-fatigue, anti-stress, anti-oxidative, and anti-aging effects. 20(S)-Protopanaxadiol (PPD), a intestinal metabolite of ginsenosides, is one of the active ingredients in ginseng. In this study, we investigated the synergistic anticancer effect of 20(S)-PPD and temozolomide (TMZ) and the mechanism of 20(S)-PPD on glioblastoma cells. Methods and Results : We examined cell viability and the morphological changes of C6 cells after treatment of 20(S)-PPD and TMZ. 20(S)-PPD showed a potent antiproliferative activity against C6 cells by triggering apoptosis. 20(S)-PPD-induced apoptosis was characterized by a dose-dependent mitochondrial damage. 20(S)-PPD and TMZ had a synergistic effect in increasing mitochondrial damage via caspase 3 activation. Conclusion : These results revealed an unexpected mechanism of 20(S)-PPD and TMZ, triggering a mitochondrial-mediated apoptosis in C6 cells. Our findings encourage further studies of 20(S)-PPD as a promising chemopreventive agent against glioblastoma.

Cleft palates with or without cleft lip is one of the most common congenital craniofacial defects in dogs. It has been reported that monogenic autosomal recessive inheritance caused this defect in this species. However, here, we aimed to report cleft palate in a cloned dog. A fibroblast cell line was established from skin tissues of an eight-year-old German shepherd dog. Blood was collected from oocyte donor dogs, and serum progesterone concentration was measured by chemiluminescence enzyme immunoassay method. Ovulation was determined when serum progesterone results reached 5-10 ng/ml, and in vivo matured oocytes were collected surgically about 72 hr after ovulation. Donor cells were cultured with Dulbecco’s modified Eagle medium supplemented with 10% (v/v) fetal bovine serum until confluence. An in vivo matured oocyte was enucleated, and a donor cell was injected into the perivitelline space. The oocyte-cell couplet was electrically fused, and chemically activated. Reconstructed embryos were transferred to an oviduct of a recipient. Pregnancy diagnosis was performed 27 days after the embryo transfer, and ultrasonography of fetal heart beat, and rectal temperature and serum progesterone value of recipient was monitored until the day of delivery. Microsatellite analysis was performed using genomic DNA of cell donor, clones, and oocyte donors. As results, a total of 74 cloned embryos were transferred to five recipients, and one recipient diagnosed as pregnant with two fetuses by ultrasonography and radiology. Caesarean section was performed on day 58 after embryo transfer due to a decreased heart beat of a fetus, which was lower than 180. Two cloned puppies with 640g and 320g of birth weight were delivered safety, but the small one was born with a cleft palate. Microsatellite analysis results of both clones were identical with the cell donor. Cleft palate of the clone was surgically corrected on day 40 after birth. To our knowledge, there has been no report about cleft palate in cloned dogs, and also, no report about clones with different phenotype of cleft palate in dogs. Therefore, this study can give a clue of cleft palate in dogs, which might not be a genetic cause. Further studies about aberrant epigenetic reprogramming in those clones are needed.