Human embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo. Human ES cells have the capacity to differentiate into various types of cells in the body. Human ES cells are indefinite source of cells for cell therapy in various degenerative disorders including neuronal disorders. Directed differentiation of human ES cells is a prerequisite for their clinical application. The objective of this study is to develop the culture condition for the derivation of neural precursor cells from human ES cells. Neural precursor cells were derived from human ES cells in a stepwise culture condition. Neural precursor cells in the form of neural rosette structures developed into neurospheres when cultured in suspension. Suspension culture of neurospheres has been maintained over 4 months. Expressions of nestin, soxl, sox2, pax3 and pax6 transcripts were upregulated during differentiation into neural precursor cells by RT-PCR analysis. In contrast, expression of oct4 was dramatically downregulated in neural precursor cells. Immunocytochemical analyses of neural precursor cells demonstrated expression of nestin and SOX1. When induced to differentiate on an adhesive substrate, neuro-spheres were able to differentiate into three lineages of neural systems, including neurons, astrocytes and oligo-dendrocytes. Transcripts of sox1 and pax6 were downregulated during differentiation of neural precursor cells into neurons. In contrast, expression of map2ab was elevated in the differentiated cells, relative to those in neural precursor cells. Neurons derived from neural precursor cells expressed NCAM, Tuj1, MAP2ab, NeuN and NF200 in immunocytochemical analyses. Presence of astrocytes was confirmed by expression of GFAP immuno-cytochemically. Oligodendrocytes were also observed by positive immuno-reactivities against oligodendrocyte marker O1. Results of this study demonstrate that a stepwise culture condition is developed for the derivation of neural precursor cells from human ES cells.

High-quality and high-phytonutrient watermelon fruits have strong market opportunities besides their health related benefits. Hence, investigating quality and nutritional related traits of watermelon genetic resources could provide important baseline data in breeding for increased lycopene content thereby increasing the marketability of watermelon. To this end, we have examined some fruit morphological traits and lycopene content of 105 genetic resources. Seeds, originally obtained from 22+ countries, were obtained from the National Agrobiodiversity Center, Jeonju, South Korea, grown in an experimental field and harvested at a fully mature stage. The size of pistil scar (SPS), the width of stripes (WS), weight of fruit (WF), length of fruit (LF), width of fruit (WIF), the thickness of pericarp (TP), soluble solids content (SSC), fruit shape in longitudinal section, ground color of skin, the intensity of the green color of skin, fruit shape at the apical part, grooving distribution, conspicuousness of stripes, and main color of the flesh were recorded on the field and inside laboratory and the lycopene was measured using spectrophotometric and HPLC methods. Watermelon fruits have shown a diverse morphological characters. Red and pink fleshed fruits dominated in the entire collections. Fruits with higher thickness of rind were found to exhibit less soluble solid content (SSC). Korean origin fruits were characterized by intermediate SSC while the United States of America (USA), Russia (RUS), Tajikistan (TJK), Turkmenistan (TKM), Taiwan (TWN), and Uruguay (URY) originated fruits had the highest SSC. The lycopene content varied between 41.37 and 182.82 ㎍/g, 2.81 and 163.72 ㎍/g, and 3.54 and 255.47 ㎍/g using HPLC, UV-Vis spectrophotometer, and microplate reader spectrophotometer, respectively. Red- and pink-fleshed fruits had the highest levels of lycopene content compared to the yellow- and orange-fleshed. Lycopene content had a significant positive correlation with SSC, however, no correlations were detected between lycopene and other quantitative fruit morphological characters. Our study demonstrated high diversity exists in fruit morphological traits and lycopene content of the germplasm collections which provide beneficial baseline data for a future breeding program and utilization of watermelon germplasm collections in gene banks for the maintenance and improvement of the current levels of production, marketability, and health-related benefit of watermelon fruits.

Aralia cordata (A. cordata), which belongs to Araliaceae, is a perennial herb widely distributed in East Asia. We evaluated the anti-inflammatory effect of stems (AC-S), roots (AC-R) and leaves (AC-L) extracted with 100% methanol of A. cordata and elucidated the potential signaling pathway in LPS-stimulated RAW264.7 cells. The AC-L showed a strong anti-inflammatory activity through inhibition of NO production. AC-L dose-dependently inhibited NO production by suppressing iNOS, COX-2 and IL-β expression in LPS-stimulated RAW264.7 cells. AC-L inhibited the degradation and phosphorylation of IκB-α, which donated to the inhibition of p65 nuclear accumulation and NF-κB activation. Furthermore, AC-L suppressed the phosphorylation of ERK1/2 and p38. These results suggested that AC-L may utilize anti-inflammatory activity by blocking NF-κB and MAPK signaling pathway and indicated that the AC-L can be used as a natural anti-inflammatory drugs.

Background : Scopolamine induces cholinergic dysfunction and oxidative stress, and the impairment of memory function. Therefore, oxidative stress and cholinergic dysfunction are important role of the brain pathology of amnesia. In this study, we investigated the impact of Safflower seed against oxidative stress and cholinergic dysfunction on scopolamine-induced amnesic mice. Methods and Results : Mice were orally pretreated with safflower seed (100 ㎎/㎏ body weight) or vehicle for 7 days, and scopolamine (1 ㎎/㎏ body weight) was injected intraperitoneally, 30 min before the behavior tests such as T-maze and novel objective recognition test on first day. To evaluate learning and memory function, the Morris water maze task was performed for 5 days, consecutively. The results showed that spatial perceptive ability and novel object recognition was significantly increased by preadministration of safflower seed compared with scopolamin-induced control mice in the behavior tests. Consistently, immuno blot revealed the elevated expression of superoxide dismutase 1 in the safflower seed pretreated mice, compared to the control mice. Moreover, protein expression of acetylcholinesterase was decreased in safflower seed pre-treated group. Conclusion : Subsequently, our results suggests that the Safflower seed extract improved memory impairment through inhibition of cholinergic dysfunction and oxidative stress.

본 연구는 완주에서 재배된 세계 주요 11개 적포도 품종으로 제조된 적포도주의 향기 성분을 headspace-solid phase microextraction 분석법으로 확인하였다. 향기성분은 총 75종이 확인되었다. 아로마화합물은 그들의 OAV 값에 의해 5 그룹으로 나뉘었다. 알콜, 알데하이드, 에스테르, C6 화합물이 11개 적포도주의 주요한 향기성분이었다. Isoamy alcohol 알콜과 phenylethyl 알콜은 11개 포도주에서 공통적으로 꽃향기, 달콤한 향을 나타내는데 중요한 물질이었다. Octanoic acid, ethyl ester, hexanoic acid ethylester은 모든 레드와인에서 과실향과 꽃향, 달콤한 향을 내는 중요한 성분이었다. 1-Hexanol은 모든 포도주에서 분석되었으나 풀향을 나타내는 향으로 나타났다. Chanceller, Malbec, marchel, Nsrsha, Pinot Meunier, Sangiovetto 포도주의 주요 향기성분은 과실 향인 것으로 나타났으며 Cabernet Franc, Cabernet Sauvignon, Sauvignon Vert 포도주의 주요 향기성분은 풀향인 것으로 조사되었다. 또한, MBA와 Narsha 포도주의 경우 꽃향이 주요 향인 것으로 조사되었다. 본 연구 결과를 바탕으로 적포도주용 품종을 육성할 때 선발기준으로 향기성분 분석을 활용할 수 있을 것으로 판단되었다.

The study compared the juice and wine odorants of the Cheongsoo grape cultivar with those of Chardonnay and Reisling wines. The volatile compounds of the three grape varieties were analyzed using headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry. The most common volatile compounds in the juices from the three cultivars were terpenes, C13-norisoprenoids, etones, alcohols, and aldehydes. Terpenes were established as the most abundant group of volatiles in Cheongsoo grape juice, where as aldehydes predominated in Chardonnay and Riesling juices. Forty-two volatile compounds (acids, alcohols, esters, and others) were detected in the three white wines. The concentration of esters was about four times higher in Cheongsoo wine than in Chardonnay and Riesling wines. Five esters found in the Cheongsoo wine, namely, isoamyl acetate, ethyl butanoate, ethyl hexanoate, ethyl octanoate, and ethyl decanoate, exhibited high odor activity values (OAV) of >1. Furthermore, only Cheongsoo wine had a high OAV for isoamyl acetate odorant, which is associated with banana and sweet aromas. Therefore, the abundant and varied esters are believed to be key volatile fruity/sweet odorants in Cheongsoo wine.

Background: Glycyrrhiza uralensis Radix (GR) is a crude drugs used in Asian countries that has been reported to prevent the progression of neurodegenerative diseases such as Alzheimer’s disease. The present study examined whether GR and its active compounds, glycyrrhizic acid (GA) and isoliquiritigenin (IL), exerted protective effects on H2O2-induced oxidative damage in C6 glial cells. Methods and Results: We exposed C6 glial cells to hydrogen peroxide (H2O2) for 24 h and investigated the cellular response to GR and its active compounds by evaluating cell viability, reactivie oxygen species (ROS) production, and apoptosis-related protein expression. GR successfully mitigated the reduced cell viability and ROS production induced by H2O2 in C6 glial cells, IL and GA significantly increased the cell viability and decreased ROS production. In addition, IL and GA down-regulated apoptotic Baxdependent caspase-3 activation, but each compound exerted different mechanisms, i.e., IL dose-dependently decreased ROS production and, GA up-regulated anti-apoptotic Bcl-2 expression. Conclusions: These results demonstrated that GR and its active components, IL and GA, exhibit potential for use as natural neurodegenerative agents for the modulation of apoptosis in C6 glial cells.

Platycodon grandiflorum (Bell flower) is an important plant that has traditionally been used as herbal medicine for the treatment of cough, phlegm, sore throats, lung abscesses, chest pains, dysuria, and dysentery. The present study was initiated to investigate the feasibility of inducing shoot and root organogenesis in cultured explants of P. grandiflorum in a range of culture media and through use of various plant growth regulators (PGRs). The plantlets (Stem containing one node) were isolated and cultured on different concentrations of Murashige and Skoog (MS) medium supplemented with PGRs. We found that proliferation and elongation of shoots and roots could be achieved on ¼ MS for P. grandiflorum with wild and green petals and on ⅛ MS for P. grandiflorum with double petals. The highest levels of development and elongation of adventitious shoots and roots were observed when petal explants were cultured on ¼ MS (pH 3.8) supplemented with 5% sucrose. Increasing the agar concentration reduced shoot growth and rooting potential; nevertheless, the highest number of shoots and roots was observed on 0.6% agar. In the case of growth regulators, ¼ MS supplemented with 1 mg L-1 6-benzylaminopurine (BA) was found to be best for shooting, although higher concentrations of BA tended to reduce shoot and root elongation. The highest number of shoots was achieved on 0.5 mg ․ L-1 thidiazuron (TDZ) from double petal explants grown on ⅛ MS. However, root and shoot elongation were found to decrease when TDZ concentrations were increased. Low concentrations of kinetin, naphthalene acetic acid, indole acetic acid, and 3-indole butyric acid induced shoot and root proliferation and elongation. Taken together, our study showed that low concentrations of PGRs induced the greatest root formation and elongation, showing that the optimal concentration of PGRs for shoot proliferation was species-dependent.

Background : Despite the presence of various bioactive compounds in ginseng, there is lack of study on the variations of bioactive compounds in ginseng according to the cultivation of soil and the applied fertilizer types (or amount). Therefore, this study aims to examine the variations of 37 fatty acids (FA) and 8 vitamin E (Vit-E) vitamers in 6-year-old ginseng root cultivated in different soil types with different fertilizers regimes. Methods and Results : The profiling of 37 FAs and 8 Vit-E vitamers in 6-year-old ginseng roots was measured by gas chromatography coupled with a flame ionization detector, and then these results were statistically analyzed with chemometrics. The FA and Vit-E content in ginseng roots varied significantly with respect to soil cultivation conditions due to organic fertilizer types and amounts used. Unsaturated FA in ginseng is approximately 2.7 fold higher than the saturated FA. Linoleic, palmitic, and oleic acids were the most abundant FAs found in the ginseng roots. Also, the major Vit-E vitamer found in ginseng root is α -tocopherol. In particular, the application of rice straw compost or food waste fertilizer was increased to create nutritionally-desirable FAs and bioactive Vit-E in ginseng root. In addition, phytonutrient profiling coupled with chemometrics can be used to discriminate the cultivation conditions of ginseng. Conclusion : This preliminary study extends our understanding about the variations of FA and Vit-E in ginseng root depending on cultivation conditions. Hence, these results can be useful as basic information for reliable ginseng production containing high amounts of phytonutrients in a paddy-converted field.

Background : Water uptake and flow across cellular membranes is a fundamental requirement for plant growth and development, and plant water status is important not only for plant growth under favorable conditions but also for ability of a plant to tolerate adverse environmental conditions. Thus identification of plasma membrane water channel genes (aquaporins) in ginseng provides extensive information for functional studies and the development of markers for salinity stress tolerance. Methods and Results : For salinity treatment, the plants were grown for 4 weeks in culture medium gelled with 0.8% Phytoagar, and the old media were replaced with the fresh medium containing NaCl at 0, 50, 100, 200 and 400 mM, respectively. The samples for stress treated and non-stressed plants were collected from 6h to 72h, and frozen immediately into liquid nitrogen. According to the sequence information from the assembled transcripts, four primer pairs were designed from the aquaporin gene regions. In order to determine the pattern of aquaporins expression in ginseng seedlings to salinity stress, we conducted semi-quantitative RT-PCR. Conclusion : A tonoplast intrinsic protein 1 (TIP1)-type aquaporin is not only believed to be essential for plant life, but also to be beneficial for growth under salinity stress. Therefore, a deeper understanding of aquaporin genes in ginseng will be essential for crop improvement, which could help us to understand the molecular genetic basis for the ginseng genetic improvement and also provide the functional genetic resources for selective breeding and transgenic research.

Background : This study was carried out to understand the effect of seedling weight (SW) on growth and flowering in Panax ginseng. Methods and Results : The testing materials were Chunpoong (CP), Yunpoong (YP) and Jakyeongjong (JK). The increase of seedling (1yr) weight led to an increase in ratio of flowering plant and in number of flower per plant. The seed setting rate of two year-old plant (CP, YP, JK) increased with increase of SW at the planting time (PT) and number of flower per plant of three year-old plant (CP, YP) increased also. In the two year-old plant (JK), the ratio of three leaves per plant was 8.8, 19.6, 31.0, 42.0, 44.7 and 58.2%, respectively, in the SW of >0.6, 0.6~0.8, 0.8~1.0, 1.02~1.2, 1.2~1.4 and 1.4g<. The growth of ginseng plant was good with increase of SW at the PT. Conclusion : There was a highly positive correlation between seedling weight and flowering characteristics.

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

We estimated the orbit of the Communication, Ocean and Meteorological Satellite (COMS), a Geostationary Earth Orbit (GEO) satellite, through data from actual optical observations using telescopes at the Sobaeksan Optical Astronomy Observatory (SOAO) of the Korea Astronomy and Space Science Institute (KASI), Optical Wide field Patrol (OWL) at KASI, and the Chungbuk National University Observatory (CNUO) from August 1, 2014, to January 13, 2015. The astrometric data of the satellite were extracted from the World Coordinate System (WCS) in the obtained images, and geometrically distorted errors were corrected. To handle the optically observed data, corrections were made for the observation time, light-travel time delay, shutter speed delay, and aberration. For final product, the sequential filter within the Orbit Determination Tool Kit (ODTK) was used for orbit estimation based on the results of optical observation. In addition, a comparative analysis was conducted between the precise orbit from the ephemeris of the COMS maintained by the satellite operator and the results of orbit estimation using optical observation. The orbits estimated in simulation agree with those estimated with actual optical observation data. The error in the results using optical observation data decreased with increasing number of observatories. Our results are useful for optimizing observation data for orbit estimation.

To understand molecular mechanisms underlying adaptation of plant cells to saline stress and stress memory, we developed Arabidopsis callus suspension-cultured cells adapted to high salt. Adapted cells to high salt exhibited enhanced tolerance compared to control cells. Moreover, the salt tolerance of adapted cells was stably maintained even after the stress is relieved, indicating that the salt tolerance of adapted cells was memorized. Salt-adapted and stress memorized cells were densely aggregated and formed multi-layered cell lump. Cell morphology analysis using transmission electron microscopy indicated that cell wall thickness of salt-adapted cells was significantly induced compared to control cells. In order to characterize metabolic responses of plant cells during adaptation to high salt stress as well as stress memory, we compared metabolic profiles of salt-adapted and stress-memorized cells with control cells by using NMR spectroscopy. A principle component analysis showed clear metabolic discrimination among control, salt-adapted and stress-memorized cells. Compared with control cells, metabolites related to shikimate metabolism such as tyrosine, and flavonol glycosides, which are related to protective mechanism of plant against stresses were largely up-regulated in adapted cell lines. Moreover, coniferin, a precursor of lignin, was more abundant in salt-adapted cells than control cells. The results provide new insight into metabolic level mechanisms of plant adaptation to saline stress as well as stress memory.