Fetal bovine serum (FBS) is the most frequently used serum for the cultivation of mammalian cells. However, since animal-derived materials might not be appropriate due to safety issues, allogeneic human serum (HS) has been used to replace FBS, particularly for the culture of human cells. While there has been a debate about the advantages of HS, its precise effect on human adult stem cells have not been clarified. The present study aimed to investigate the effect of HS on the human eyelid adipose stem cells (HEACs) in vitro. When HEACs were cultivated in a medium containing 10% HS, many cells moved into several spots and aggregated there. The phenomenon was observed as early as 9 days following 10% HS treatment, and 12 days following 5% HS plus 5% FBS treatment. However, the aggregation was never observed when the same cells were cultivated with 10% FBS or bovine serum albumin. To examine whether cell density might affect the aggregation, cells were seeded with different densities on 12-well dish. Until the beginning of aggregation, cells seeded at low densities exhibited the longest culture period of 16 days whereas cells seeded at high densities showed the shortest period of 9 days to form aggregation. The number of cells was as the least for the low density group, and as the greatest for the high density group. When human cord blood serum or normal bovine serum was examined for the same effect on HEACs, interestingly, cord blood serum induced the aggregation of cells whereas bovine serum treatment has never induced. When cells were cultivated with 10% HS for 9 days, they were obtained and analyzed by RT-PCR. Compared to FBS-cultivated HEACs, HS-cultivated HEACs did not express VIM, and less expressed GATA4, PALLD. On the other hand, HS-cultivated HEACs expressed MAP2 more than FBS-cultivated HEACs. In conclusion, human adult stem cells could move and form aggregates by the treatment with human body fluids.

Early growth response 1 (Egr1) is an immediate early response gene which is induced by various external stimuli and acts as transcription factor to direct second-wave gene expression leading to cell growth, differentiation and/or apoptosis. It is well known that Egr1 regulates transcription of a cluster of genes in cancers and luteinizing hormone (LH) beta subunit in the pituitary. In addition to function of Egr1 in cancers and pituitary, we recently showed that Egr1 acts as a local master regulator to mediate estrogenic actions in the uterus. However, regulatory mechanism by which Egr1 directs transcription of its downstream target genes in the uterus remains to be yet explored. Thus, we have tried to identify direct target genes of Egr1 in the uterus by analyzing mRNA microarray data sets followed by in silico promoter analyses with chromatin immunoprecipitation (CHIP). mRNA expression profiles of Egr1(－/－) uteri and Egr1(－/－) ovaries were compared to those of wildtype mice to provide a potential list of direct target genes of Egr1 in the uterus. Whereas Egr1 is rapidly and transiently induced in the ovary and the uterus by external stimuli, LH and estrogen, respectively with a similar manner, a list of differentially expressed genes between Egr1(+/+) and Egr1(－/－) mice were barely overlapped between these two datasets. This result suggests that the transcriptional network of Egr1 in the uterus is quite different from that in the ovary. The list of differentially expressed genes in Egr1(－/－) uterus was enriched by RT-PCR. In silico analyses with MatInspector provided evidence that Egr1 binding sites are relatively enriched in －500 bp promoter regions of genes in the list. CHIP assays for Egr1 antibody with uterine tissues 2 h after estrogen treatment reinforced the possibility that genes identified in this study such as Gadd45g and Lbh could be directly regulated by Egr1 in uterine context. Collectively, we show that bioinformatic analyses of expression profiles with in silico analyses could be a useful tool to enrich potential candidates of direct target genes of transcription factors.

Early growth response 1 (Egr1) belongs to the Egr family of zinc finger transcription factors that regulates cell growth, differentiation, and apoptosis. Egr1(－/－) female mice are infertile due to anovulation resulting from luteinizing hormone β subunit (LHβ) deficiency. While it is clear that Egr1 is a critical factor to regulate transcription of LHβ in the pituitary gland, function of Egr1 and mechanisms by which estrogen (E2) and/or progesterone (P4) regulates Egr1 in uterus still remain unexplored. Using multiple approaches, here we have characterized regulatory mechanism of Egr1 induction in the uterus and uterine phenotypes of Egr1(－/－) mice. Eight-week-old female mice were ovariectomized (OVX) and rested for a week. Uteri of OVX mice treated with various concentrations of E2 and/or other hormones were collected at 2h after hormone treatment unless otherwise indicated. Collected uteri were utilized for mRNA microarrays, realtime-RT-PCR, Western blotting, and histological analyses for immunofluorescence and BrdU staining. Egr1 mRNA was rapidly induced with the highest level at 2h after E2 treatment and gradually decreased to basal levels at 12 h. E2-induced phosphorylation of ERK1/2 and AKT, and Egr1 transcription were effectively inhibited by pretreatment of ICI 182,780. Pharmacological inhibition of ERK1/2, but not AKT significantly blocked E2-induced Egr1 expression in the uterus. P4 effectively dampened E2-dependent Egr1 transcription and its antagonistic effects were partially interfered with RU486 pretreatment. Interestingly, BrdU incorporation experiments provided evidence that epithelial cells undergo hyperproliferation in Egr1(－/－) mice. This is consistent with microarray data that several key factors for cell cycle progression such as cyclin Ds and E2F1 are overexpressed in these mice. Furthermore, in the uteri of OVX Egr1(－/－) mice treated with E2+P4, stromal cell proliferation is severely impaired and epithelial cells persistently proliferating. While ovulation, fertilization and embryo development normally occur in Egr1(－/－) mice treated with a superovulation regime to rescue LH deficiency, embryo implantation is severely impaired. Blastocysts were not able to implant even on day 6 of pregnancy in Egr1(－/－) mice. In addition to embryo implantation, uterine response to artificial decidualization in hormone-primed Egr1(－/－) OVX mice was relatively less than that of wildtype mice. Collectively, our results show that Egr1, which is rapidly induced by E2-ER-ERK1/2 pathways, is a critical factor to control E2-dependent cell proliferation via regulation of a spectrum of genes for embryo implantation in the uterus.

MicroRNAs (miRNAs) are single-stranded, small non-coding RNAs which are critical for gene regulatory networks by directing the translational repression or degradation of complementary target mRNAs. They can be divided into two subclasses: canonical and non-canonical miRNAs. Canonical miRNAs are produced from long primary transcripts by sequential complex events in which RNA III enzymes such as Drosha and Dicer and accessory factors such as DGCR8 and Argonautes work cooperatively. DGCR8 is a RNA-binding protein that assists Drosha to process canonical miRNAs in the nucleus. To understand function of canonical miRNAs in uterine physiology, we have characterized uterine phenotypes of uterine-specific DGCR8 knock-out mice (uDGCR8 KO, DGCR8flox/flox; PRcre/+), and hormonal regulation of expression profiles of major factors working for miRNA biogenesis in the uterus. Gross morphological and histological analyses, immunohistochemistry, PCR and realtime RT-PCR were performed. While DGCR8 and Drosha do not seem to be regulated by ovarian steroid hormones, expression of Dicer, Exportin 5 and Argonaute 2 was transiently increased by E2 but not by P4. Combination of E2+P4 did not have any additional effects on their expression profiles. Genomic DNA PCR analyses showed that while DGCR8 gene is not completely deleted in the uterus, DGCR8 is specifically deleted in the uterus where PR is strongly expressed. uDGCR8 KO females bred with fertile males did not produce any offspring, suggesting that these mice are infertile. Vaginal smear analyses provided evidence that these mice do not undergo estrous cycle. Whereas gross morphology and histological analyses of uteri of 3-week-old uDGCR8 KO mice is similar to that of DGCR8flox/flox mice (control), uteri of 5- and 8-week-old mice are extremely thinner and shorter than those of control mice. These results were supported by significant decrease in uterine weight/body weight of uDGCR8 KO mice at 5-week-old age onward. Interestingly, this phenotype is reflected by significant increase of PR expression in the uteri of 4-week-old mice. Expression of DGCR8 and Dicer is significantly increased after birth. BrdU incorporation experiments showed that cell proliferation governed by ovarian steroid hormones does not normally occur in these mutant mice. Furthermore, artificial decidualization does not occur in these mice. Collectively, these results conclude that canonical miRNAs plays critical roles in normal uterine development and steroid hormone-dependent uterine function.

It is well‐known in the literature of Korean linguistics that the difficulty with the semantic treatment of –ess lies in the fact that it can be used to refer to a past eventuality or a perfect eventuality, depending on the context of use. For this reason, after pointing out that it is of no use to posit that –ess is a past tense marker, a perfect marker, or both, this paper argues that –ess itself is neither a past tense marker nor a perfect marker, which completely departs from the previous treatments of –ess . To put it differently, -ess is indeterminate between a past tense marker and a perfect marker across the sentences in which it occurs. The context of use serves to resolve its indeterminacy by presenting the most salient semantic interpretation that may arise from –ess .

This study aims to identify the degree of safety when mariners take their actions in several different situations. We have carried out many experiments to observe mariners' behavior and then measured the safety level based on their actions to avoid dangerous situations of ships collision. One of the most important actions that mariners have to take, either as their daily routine or when they are in a collision situation and then want to avoid that situation is the lookout. In this paper, behaviors on the lookout have been defined as a standard sequence of three steps which are "time of first detection", "time of recognition as risky vessel" and "time of starting avoiding action", and the suitability and applicability of the definition have been shown. And also we propose the risk assessment on ships collision and the recommendation for reducing ships collision at sea. Some analyzing results and the application of the results are reported. By combining these knowledge and some systematic studies, we propose the risk assessment on ships collision and the recommendation for reducing ships collision at sea.

The antioxidant potential and enzyme activities in Salicornia herbacea, Corylopsis coreana, Erythronium japonicum, Phragmites communis, Momordica Charantia, Nelumbo nucifera, Salvia plebeia, Portulaca oleracea, Ficus carica, Citrus junos and Cornus officinalis were determined. Their antioxidant activities were measured using DPPH radical scavenging and nitrite scavenging activity. Enzyme activities in investigated plants were evaluated as superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX). The DPPH scavenging rate from 100 to 2500 mgL-1 was the highest in the flower of Corylopsis coreana. However, it was not detected in most of the samples at concentration below 100 mgL-1. The nitrite scavenging activity according to each kind of resource plants was significantly higher in the stem of Corylopsis coreana and leaf of Nelumbo nucifera. The root extract of Erythronium japonicum had the highest SOD enzyme activity of 94.0% while leaf of Salvia plebeia showed the lowest SOD enzyme activity of 30.4%. The activity of CAT and APX showed higher values in the stem of Corylopsis coreana, root of Erythronium japonicum and root of Phragmites communis in comparison with other plants. The activity of POD showed significantly high values in stem of Corylopsis coreana, Momordica Charantia and pericarp of Citrus junos extracts. The antioxidant enzyme activities differ significantly in different plants. In conclusion, we showed that Corylopsis coreana, Erythronium japonicum Cornus officinalis, and Momordica Charantia had the potent biological activities. Therefore, these plant resources showing antioxidant activity could be good materials for development of source of functional healthy food.

본 연구는 구강 내 방사선 촬영 시 동일조건 하에서 촬영장비의 기종을 다르게 한 경우와 동일 기종에서 노출조건을 일정하게 하였을 경우 획득되는 방사선의 선량변화를 알아보고자 하였다.장비의 기종에 따른 변화를 알아보기 위해 3대의 각기 다른 촬영 장치를 이용하여 상악 전치부, 소구치부, 대구치부의 3가지의 촬영조건을 이용하여 선량을 측정하였고 장치의 재현성 검사를 위해서 각 장치에 동일한 조건으로 3회 조사하여 선량변화를 알아보았다. 선량의 측정은 반도체 측정기를 사용하였으며, 필름의 농도값을 획득하여 진단에 유효적인 범위에 만족하는지 분석해 보았다. 같은 촬영기를 사용하여 실험한 결과 각 부위별로 선량의 차이는 최대값과 최소값이 적게는 3배 많게는 10배의 선량 차이를 보였다. 흑화도의 범위 또한 적절한 범위인 0.25∼2.0를 초과한 2.10∼2.95의 허용범위를 보였다.동일한 촬영기로 특정 부위를 같은 조건에서 촬영하더라도 측정된 선량값은 큰 차이를 보였으며 필름의 농도 분석의 결과 적합한 농도 분포를 보이지 않았던 것으로 나타났다. 이는 환자의 재촬영을 발생시킬 수 있으며 진단시 오류를일으킬 수 있어 환자의 피폭에 직·간접적인 피해를 일으킬 수 있다. 따라서 치과 방사선 촬영장치 또한 정기점검이 필요할 것으로 생각되며 표준화된 촬영조건과 현상조건이 제공되어야 한다고 사료된다.

사용후핵연료 심지층처분장 부지선정과 최종 처분장부지의 처분적합성을 평가하는 업무는 시행-착오 를 줄이고 기술적 신뢰성 확보와 합리적이고 효율적인 업무수행을 추구하여야 한다. 이에 선행하여, 우리 나라에 적용 가능한 처분장부지의 지질환경 요건 설정을 위한 기본방향과 개별 인자의 처분적합성지표를 가능한 한 정량화하여 설정하고 업무에 적용하여야 한다. 사용후핵연료 처분장부지 선정과 최종처분장 부지의 안전성확보를 위한 처분요건과 관련하여 IAEA 및 OECD 회원국들과 처분연구 및 상용사업 수행 관련 선진국가들의 사례를 바탕으로 요건 별로 구분하여 현황을 분석하였다. 여기서는 사용후핵연료 처 분장 부지로서 암석·암반이 갖추어야 할 충분 혹은 선호요건에 대한 이해 제고와 관련 세부 기술지침을 도출하는데 기여하고자 하였다. 이를 토대로 어떠한 암석·암반이 상대적으로 보다 유리한 조건을 가지 는 선호요건으로 제시해야 하는지, 그리고 충분요건과 선호요건을 적용하여 후보부지 조사·선별평가 기 간 동안 부지선정업무에 반영하고 평가하고 결정하여야 하는 방법론을 도출할 수 있도록 기본 골격을 제 시하였다. 또한 처분안전성 확보를 위해 필요한 기본적인 사항을 검토하고 서술하였다. 본 논문에서 기술 한 항목들은 처분안전성 확보를 위한 처분요건의 기술지침 구성 체계, 처분안전성 확보개념, 다중방벽 기 능 조건, 천연방벽의 지질환경 기본요건, 그리고 우리나라에 적용 가능한 처분장부지 지질환경 기본요건 (안) 등으로 구성된다. 우리나라의 사용후핵연료 심지층처분장 부지의 위치에 관한 사업자 기술지침 요건 으로 제안하였다. 이와 관련하여 충분요건과 선호요건으로, 화산활동, 지진활동, 단층운동 융기·침강 운 동 및 기후·해수면변동 등 장기지질안정성 요건을 비롯한 15개 충분요건과 48개 선호요건을 제안하였 다. 이들 요건은 우리나라의 지질환경 특성을 충분히 반영하여 후속되는 각 부문별 특성에 적합한 정량적 인 기술 기준 및 지침으로 개발되어야 할 것이다. 정량적 기술지침의 도출은 상용 처분장부지 선별평가과 정 및 처분장 부지적합성평가 과정으로부터 확립될 수 있을 것이다. 또한 다양한 부문별 안전사례(safety case) 작성 혹은 연구용 지하처분연구시설 (underground research laboratory: URL)을 이용한 처분시스 템의 실증과정 등을 통하여 객관적이고 신뢰성있는 정량적인 지침들이 확립될 수 있을 것이다.

This study aims at classifying and analyzing the vegetation structure of Castanopsis sieboldii forest, one of the evergreen broad-leaved forests found under the warm-temperate climate of Korea. It is also compared with the ones of the Castanopsis sieboldii forest in Japan where most similar such forest of Korea, to find unique vegetation structures of the only Korean forest. Vegetation structure of Korean Castanopsis sieboldii forest was divided into two units at the level of community units both of Ardisia japonica-Castanopsis sieboldii community and Ardisio-Castanopsietum sieboldii association. The association carries similar type with the vegetation system of Japan, but any subunits differentiated with the Japan were found vary much. Hierarchical cluster analysis brings in similar result with the analysis on the vegetation structure as well.

Human eyelid adipose-derived stem cells (hEAs) and amniotic mesenchymal stem cells (hAMs) are very valuable sources for the cell therapeutics. Both types of cells have a great proliferating ability in vitro and a multipotency to differentiate into adipocytes, osteoblasts and chondrocytes. In the present study, we evaluated their stem cell characteristics after long-time cryopreservation for 6, 12 and 24 months. When frozen-thawed cells were cultivated in vitro, their cumulative cell number and doubling time were similar to freshly prepared cells. Also they expressed stem cell-related genes of SCF, NANOG, OCT4, and TERT, ectoderm-related genes of NCAM and FGF5, mesoderm/endoderm-related genes of CK18 and VIM, and immune-related genes of HLA-ABC and 2M. Following differentiation culture in appropriate culture media for 2-3 weeks, both types of cells exhibited well differentiation into adipocyte, osteoblast, and chondrocyte, as revealed by adipogenic, osteogenic or chondrogenic-specific staining and related genes, respectively. In conclusion, even after long-term storage hEAs and hAMs could maintain their stem cell characteristics, suggesting that they might be suitable for clinical application based on stem cell therapy.

This experiment was conducted to clarify the antitussive effect of Fritillaria unibracteata Hsiao et K. C. Hsia. The antitussive effect in ethanol (75, 50, 30%) extracts from F. unibracteata was relatively high. The ethanol extracts of 20, 40 mg/kg concentrations were especially good antitussive effect. The ethanol extracts of F. unibracteata showed lower antitussive activities of 20~30% than sodium chloride injection used as a blank space control.