Anthocyanins are naturally occuring phytochemicals and the main components of the coloring of plants, flowers and fruits. They are known to elicit antioxidative, anti-inflammatory and cancer preventive activity. In this study, we investigated anthocyanins in black / yellow soybean seedcoats using different methods of detection - thin layer chromatography (TLC), capillary zone electrophoresis (CZE) and HPLC analysis. The anthocyanins in soybean seedcoats were extracted by five independent methods of extraction and the aglycons (anthocyanidins) of the corresponding anthocyanins were prepared by acid mediated hydrolysis. The anthocyanin / anthocyanidin in black soybean seedcoat showed characteristic TLC mobility, CZE electrophoretic retention and HPLC migration time while little of anthocyanins were detected from yellow soybean seedcoat. The extracted anthocyanins showed pH dependent retention time in CZE and spectral change in UV-Vis spectrum. HPLC analysis of the hydrolyzed extract of black soybean seedcoat identified the presence of four anthocyanidins. The major anthocyanin in black soybean seedcoat was cyanin (cyanidin-3-O-glucoside), with the relative order of anthocyanidin in cyanidin > delphinidin > petunidin > pelargonidin.

In our present study, total methanol extracts prepared from B. platyphylla var. japonica showed a significant increase in cell proliferation upon the induction of oxidative stress by hydrogen peroxide or y-ray irradiation. Total methanol extracts were fractionated into five separate preparations i.e. n-hexane, dichloromethane, ethylacetate, n-butanol and water fractions. Among these, the ethylacetate and butanol fractions of B. platyphylla var. japonica showed the highest protective effects against oxidative stress induced by hydrogen peroxide. These fractions also showed strong protective effects against y-ray irradiation. When we evaluated the cytotoxicity of these fractions, the butanol fraction showed no effects in a colony formation assay. In addition, the butanol fraction showed a cell proliferation activation effect evidenced by significant increase in the colony formation of y-ray irradiated cells. Both a radical scavenging activity and clonogenic activity assay suggested that the mechanism behind this protective effect against reactive oxygen species may be due to the radical scavenging and cell proliferation activity of B. platyphylla var. japonica extracts.

Temperature-dependent development and oviposition component models were developed for Pseudococcus cryptus Hempel (Homoptera: Pseudococcidae). Egg development times decreased with increasing temperature and ranged from 2.4d at 16℃ to 1.0d at 28℃. Total development times of nymphs reared on citrus leaves decreased from 54.9d at 16℃ to 17.4d at 28℃ and 19.3d at 32℃. As P. cryptus showed an ovoviviparous reproductive behavior, the periods of egg and the 1st nymph were combined. By fitting linear models to the data the lower developmental threshold temperatures for egg-1st nymphs, the 2nd nymphs, the 3rd nymphs, and all nymphs combined were calculated as 8.7, 12.8, 13.1, and 12.1℃, respectively. The thermal constants were 198.6, 84.7, 69.8, and 296.3 degree-days for each of the above stages. The non-linear model based on a Gaussian equation, which fits the relationship between development rate and temperature was well for all stages. Adult longevity decreased with increasing temperature and ranged from 80.4d at 16 to 31.3d at 32.0℃. Also, preoviposition and oviposition periods showed a similar pattern with the longevity. P. cryptus had a maximum fecundity of 111 eggs per female at 28℃, which declined to 102.7 eggs per female at 32℃.

This study was to taken to demonstrate the effects of exogenous nitric oxide(NO) on hu rnan pu lp cell s ‘ In volvement of cyclic 3’, 5' -monophosphate(cGMP) in p버 paJ protection induced by herne oxygenase-l (J-lO-l) against NO-induced cytotoxicity , By use of Western blotting and cell viabi lity assay, we have examined the cytotoxicity and J-lO-l induction in pulp cells that were treated with NO donor ‘ S-nitroso-N-acetyl-D, L-penici 1 lamine(SNAP) , We have assessed wheathel' HQ--l contributes the cytoprotective effect against the cytotoxicity caused by NO, and inves tigated the l'elationship between HO-l and cGMP in the s ignaling pathway, SNAP decreased cell via bility but in creased HO-l expl'ession in a concentl'ation- and time一dependent manner in hurnan pu lp cells NO-induced cyto toxicity was inhibited in the presence of the hemin(inducer of HO-l) , whel'eas was en hanced in the pl'esence zinc protoporphyrin IX(ZnPP IX, HO-l inhibitor), thus Lhe NO-induced cytoLoxicity was cOl'related with HO- l expression. R‘ etreatment with a rnemhrane-permeable cGMP analog, 8-bromo-cGMP, restored cell death and enhanced the HO-l protein expression induced by SNAP, ln contrast‘ inhibition of guanylate cyclase by lI-l -[1,2,4] ox adiazole[ 4,3 口]quinoxalin-l-one(ODQ) pretreated pulp cells to 1 mM SNAP resulting in marked cytotoxicity , These findings , demonstrating a link between J-lO-l, regulated thl'ough the cGMP system and NO-induced cytotox.icity in huma띠 p버 p ceJls , suggesti ng a protective 1'ole of HO-l in pulp infl ammatory disease

본 연구는 콘크리트 교량의 지진취약도 곡선을 개발함에 있어 성능 스펙트럼 기법(Capacity Spectrum Method)에 대한 고찰을 통해 가장 적절한 해석방법을 제시하는데 그 목적이 있다. 원래 성능 스펙트럼 기법은 빌딩 구조물을 위한 간략화된 정적 비선형 해석의 일환으로 개발되었는 바, 본 연구에서는 이 기법을 교량의 지진취약도 곡선을 개발하는데 응용하였다. 서로 다른 네가지의 방법으로 성능 스펙트럼 기법을 통해 구해진 취약도 곡선들을 비선형 시간이력해석 방법에 의해 구해진 취약도 곡선과 비교하였다. 취약도 곡선은 두 개의 변수를 가진 lognormal 분포를 따르는 것으로 가정하였으며 PGA(Peak Ground Acceleration)의 함수로 나타내어졌다. FEMA(Federal Emergency Management Agency) SAC(SEAOC-ATC-CUREe) steel 프로젝트에 의해 개발된 로스앤젤레스 지역 60개의 지진이 교량해석을 위해 사용되었다. 성능 스펙트럼 기법과 시간이력해석에 따라 만들어진 교량의 지진취약도 곡선들을 비교 검토한 바, 이 중 하나의 방법이 부합되는 결과를 보여주었다. 요구 스펙트럼 작성시 본 논문에서 제시된 지침을 따르면 비선형 시간이력 해석시와 유사한 결과를 얻을 수 있을 것으로 사료된다. 다만 지진과 교량이 지닌 특수성으로 인해 본 연구의 결과가 항상 적용되는지는 더 심도있는 연구를 통해 검증되어야 할 것이다.

흡연 유무의 남성을 대상으로 뇌 회백질의 손상 유무를 파악 할 수 있는 확산텐서영상을 검사하여 영상을 획득 한 후 Tract-Based Spatial Statics(TBSS)방법으로 뇌 회백질 부위의 기저핵 신경섬유로의 비등방도 FA(fractional anisotropy)값을 측정 분석한 결과 모든 영역에서 흡연자가 비흡연자보다 비등방성 측정값이 낮게 관찰되었으며 FA값은 통계적으로 유의하였다. 본 연구의 측정한 FA결과 값으로 추측하자면 즉, 흡연이 뇌 회백질 기저핵의 모든 해부학적 미세 구조성 변화에 크게 영향을 미치며 신경 섬유로를 손상시키고 이와 관련된 기능적 이상에 영향을 준다고 할 수 있다.

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.

Hepatocytes and hepatic progenitors derived from human ES cells may be a useful source for clinical application. Therefore, identification and purification of these cell types would be following important issues. There are very few candidate surface markers that can be used to identify and purify hepatic progenitor cells. In addition, indocyanine-green can be uptaken by mature hepatocytes, but cannot be applied for fluorescence activated cell sorting (FACS) due to its long emission wavelength. In the present study, we tested EpCAM as a potential marker for magnetic-activated cell sorting (MACS) of hepatic progenitors and also modified indocyanine-green into fluorescent indomonocarbocyanine for FACS-mediated sorting of mature hepatocytes after differentiation of human ES cells. Hepatic progenitor cells were sorted by MACS after incubation with anti-human EpCAM antibodies. After the final differentiation, the differentiated cells and mouse primary hepatocytes (control group) were incubated with indomonocarbocyanine and were sorted by FACS. MACS and immunocytochemistry data showed that approximately 45% of differentiated cells were EpCAM-positive cells. EpCAM-positive cells expressed α-fetoprotein, FOXa2, HnF4a, and CK18. Differentiation efficiency into albumin-positive cells was significantly higher in EpCAM-positive cells, compared to EpCAM-negative cells. Importantly, indomonocarbocyanine successfully stained cells that expressed ALB. Furthermore, FACS analysis data showed that the purity of hepatocytes that expressed albumin was significantly increased after purification of indomonocarbocyanine-positive cells. Our data demonstrated that human ES cell-derived hepatic progenitors can be efficiently isolated by MACS using EpCAM antibody. In addition, we also showed that indomonocarbocyanine can be successfully used to identify and purify mature hepatocytes using FACS.

Highly homogeneous and functional stem cell-derived hepatocyte-like cells (HLCs) are considered a promising option in the treatment of liver disease and the development of effective in vitro toxicity screening tool. However, the purity of cells and expression and/or activity of drug metabolizing enzymes in stem cell-derived HLCs are usually too low to be useful for clinical or in vitro applications. Here, we describe a highly optimized differentiation protocol, which produces more than 90% albumin-positive HLCs with no purification process. In addition, we show that hepatic enzyme gene expressions and activities were significantly improved by generating three-dimensional (3D) spheroidal aggregate of HLCs. The 3D differentiation method increased expressions of nuclear receptors that regulate the proper expression of key hepatic enzymes. Furthermore, a significantly increased hepatic functions such as albumin and urea secretion were observed in 3D hepatic spheroids and HLCs in the spheroid exhibited morphological and ultrastructural features of normal hepatocytes. Importantly, we show that repeated exposures to xenobiotics facilitated the functional maturation of HLC, as confirmed by increased expression of genes for drug metabolizing enzymes and transcription factors. In conclusion, the 3D culture system with repeated exposures to xenobiotics may be a new strategy for enhancing hepatic maturation of stem cell-derived HLCs as a cell source for in vitro high-throughput hepatotoxicity models.

Hepatocyte-like cells (HLCs) derived from human pluripotent stem cells have received extensive attention in the development of drug screening and toxicity testing. However, it has been reported that stem cell-derived HLCs showed hepatic functions that were too limited to be of use in drug screening and toxicity testing, possibly due to the lack of sufficient intercellular communication under conventional two-dimensional (2D) culture conditions. Therefore, a 3D differentiation system may overcome the in vitro limitation of 2D culture to produce stem cell-derived hepatocytes with mature metabolic functions. In this study, the feasibility of using a silicone-based spherofilm, specifically designed to produce spherical cell clusters, to generate uniformly sized 3D hepatic spheroids from hESCs was investigated. Hepatic spheroids generated on the spherofilm showed more homogenous size and shape than those generated in conventional low-attachment suspension culture dishes. Results of immunohistochemical analysis showed that expression of the mature hepatic marker albumin (ALB) increased over time during the hepatic maturation process. Furthermore, the 3D culture system mimicked the in vivo 3D microenvironment. Laminin, which is an important component of hepatic ECM, was expressed in hepatic spheroids. The results of immunohistochemical analysis indicated that the 3D culture environment is capable of generating an in vivo-like microenvironment. In addition, quantitative PCR analysis showed that the mature hepatic marker ALB and cytochrome P450 (CYP) enzymes CYP3A4 and CYP3A7 were expressed at higher levels in 3D culture than in 2D culture. This indicates that the 3D culture system is suitable for hepatic maturation and that our size-controlled 3D culture conditions might accelerate hepatic function. These results suggest that 3D hepatic spheroids significantly enhance metabolic maturation of hepatocytes derived from hESCs

MFG-E8 (Milk fat globule-epidermal growth factor VIII), also called lactadherin or BA46, SED1 is a glycoprotein found in milk and mammary epithelial cells, it is a major protein component associated with milk fat globule membrane. Previously, our study showed that expression of MFG-E8 is gradually increased with hepatic differentiation of human embryonic stem cells (hESCs). Therefore, we hypothesized that MFG-E8 would be an early cancer stem cell marker, which may predict cancer progression. Our results showed that MFG-E8 was expressed in various human cancer cell lines such as HepG2, Hep3B, and Huh7. Production and secretion of the MFG-E8 were also confirmed in the conditioned media of those three cell lines using enzyme-linked immunosorbent assay. Next, we analyzed the MFG-E8 expression in 11 clinical cases of cholangiocellular carcinoma (CC) and 33 cases of hepatocellular carcinoma (HCC) by immunohistochemistry and examined the potential correlation with β-catenin and AFP, which are known cancer markers. According to hitological criteria, the progression of HCC and CC was evaluated and classified into high, low, metastatic, and well-, moderate-, poor-differentiated, respectively. Statistical analysis indicated that incidence of both HCC and CC is significantly associated with male compared to female (P<0.05). Tumor size also has positive correlation with age (r2=08948). Our immunohistochemistry data showed that MFG-E8 was expressed both HCC and CC tissue. Interestingly, the MFG-E8 expression was significantly increased with cancer progression (P<0.05) in both cases. Additionally, b-cateninexpression was increased and its localization was changed from membrane to cytoplasm and nucleus with the degree of HCC. Likely b-catenin, AFP was also increased with the degree of HCC but it was not correlated with severalty of CC. Importantly, both AFP and b-catenin were highly co-localized with MFG-E8 in HCC. These results suggest that MFG-E8 may have important physiological roles and its expression in HCC and CC would be considered as an important prognostic factor.

Compared to wide ranges of genetic variation of natural populations, very limited Miscanthus cultivar has been released. This study was the first report on the development of Miscanthus cultivar by means of radiation breeding. Seeds of M. sinensis were initially exposed to gamma rays of 250 Gy for 24 h, generated from a 60Co gamma-irradiator. The irradiated seeds were sown and then the highly tiller-producing mutants were selected for this study. Biomass-related parameters including tiller number, plant height, stem diameter, and leaf number were measured. Ploidy level and internal transcribed spacer (ITS) were investigated to characterize the mutants compared to wild type (WT) Miscanthus. Plant height and tiller number were negatively related, where multi-tillering mutants were relatively short after 4 month growth. However stem diameter and leaf number were greater in mutants. All the materials used in this study were diploid, implying that the mutants with greater tiller numbers and stem diameter were not likely related to polyploidization. Based on the sequence of ITS regions, the mutants demonstrated base changes from the gamma irradiation where G+C content (%) was decreased in the ITS1, but increased in ITS2 when compared to WT sequence. ITS2 region was more variable than in ITS1 in the mutants, which collectively allows identification of the mutants from WT. Those mutants having enhanced tillers and allelic variations might be used as breeding materials for enhanced biomass-producing Miscanthus cultivars.

Influences of different seeding dates on growth, seed yield, fatty acid composition and oil content were investigated in flax plants for two years. The results indicated that plant height in early seeding date was higher than that of delayed seeding dates during first season. Furthermore, seeding date also significantly affected the ripened seed rate and the rate increased with the delay in seeding date in first season. Seed yield in the first crop season was significantly higher than the second crop season. Palmitic acid showed variation in different seeding dates. Contrarily, stearic acid was stable and did not changed by different seeding dates. Linolenic acid was found in highest amount in all seeding dates consecutively in two cropping years. Highest oil content was recovered from the seeds of flax sown at 29 Apr. and May 9 in first and second cropping year respectively.

Estrogens are ubiquitous signaling molecules that influence nearly every cell type, and exert profound effects on embryonic development, and differentiation. Wnt pathway, which recruits β-catenin into nuclei, and activates The Wnt-dependent transcription factors, also plays an important role in embryonic development and stem cell maintenance, and differentiation. Accumulating evidences indicate that potential convergence between these two pathways in carcinoma cells. However, physiological roles of estrogens in development and differentiation of human embryonic stem cells (hESCs) are relatively unknown. Here, we demonstrated that estrogenic compounds 17α-ethinylestradiol (EE2) and genistein (GEN) significantly increased β-catenin expression in undifferentiated hESCs cultured in feeder-free media. Interestingly, GEN treatement induced an increased trend of mesendodermal gene expressions, and significantly inhibited ectodermal gene expressions (Nestin and Pax6) in embrioid body (EB). Expectantly, GEN increased epithelial-mesenchymal transition (EMT) related gene expression (Snail2, and Twist), whereas decreased E-cadherin on day 6 of EB development. Taken together, these suggest that estrogens may in part the powerful effects on normal hESC differentiation. Mechanistic studies of estrogen signaling continue to suggest novel drug targets for stem cells and will also improve screening methods of developmental toxicity.