The use of high throughput screening (HTS) in drug development is principally for the selection new drug candidates or screening of chemical toxicants. This system minimizes the experimental environment and allows for the screening of candidates at the same time. Umbilical cord-derived stem cells have some of the characteristics of fetal stem cell and have several advantages such as the ease with which they can be obtained and lack of ethical issues. To establish a HTS system, optimized conditions that mimic typical cell culture conditions in a minimal space such as 96 well plates are needed for stem cell growth. We have thus established a novel HTS system using human umbilical cord derived-mesenchymal stem cells (hUC-MSCs). To determine the optimal cell number, hUC-MSCs were serially diluted and seeded at 750, 500, 200 and 100 cells per well on 96 well plates. The maintenance efficiencies of these dilutions were compared for 3, 7, 9, and 14 days. The fetal bovine serum (FBS) concentration (20, 10, 5 and 1%) and the cell numbers (750, 500 and 200 cells/well) were compared for 3, 5 and 7 days. In addition, we evaluated the optimal conditions for cell cycle block. These four independent optimization experiments were conducted using an MTT assay. In the results, the optimal conditions for a HTS system using hUC-MSCs were determined to be 300 cell/well cultured for 8 days with 1 or 5% FBS. In addition, we demonstrated that the optimal conditions for a cell cycle block in this culture system are 48 hours in the absence of FBS. In addition, we candidates using our HTS system which demonstrates the feasibility if using hUC-MSCs for this type of screen. Moreover, the four candidate compounds can be tested for stem cell research application.

The generation and application of porcine iPSCs (piPSCs) as a large animal model may enable the test for the efficacy and safety of the therapy in the field of human regenerative medicine. Here, we report the generation of piPSC from wild (a 10-day-old Massachusetts General Hospital miniature pig; MGH minipig) and genetically modified pig, alpha1,3-Galactosyltransferase knock-out (—/—) (GalT KO homo) and human CD46 (membrane cofactor protein) knock-in (hCD46 KI) MGH minipig (a 10-day-old). Fibroblasts were isolated from the ear skin of wild and MGH minipigs, respectively. After 2 passages, each of fibroblasts was transduced with cocktail of 6 human factors (POU5F1, NANOG, SOX2, C-MYC, KLF4, and LIN28) and cultured on a mitotically inactive mouse embryonic fibroblast (MEF) monolayer. Both of reprogrammed somatic cells expressed the classical pluripotency markers (POU5F1, NANOG, and SOX2) and surface marker (SSEA1). Similar to mouse ESCs, both piPSCs from wild and transgenic minipigs were negative for SSEA3, Tra-1-60, and Tra-1-81. Further these cells could form embryoid body (EB) and differentiate into 3 germ layers in vitro (ectoderm: FOXJ3 and PAX6, mesoderm: HAND2, and endoderm: SOX17 and GATA6). Our piPSCs may provide useful source as a large animal model for studying approaches that can reduce an immune- rejection of cell or organ transplantation.

The Fe-based self-fluxing alloy powders and TiC particles were ball-milled and subsequently compacted and sintered at various temperatures, resulting in the TiC particle-reinforced Fe self-fluxing alloy hybrid composite, and the microstructure and micro-hardness were investigated. The initial Fe-based self-fluxing alloy powders and TiC particles showed the spherical shape with a mean size of approximately 80 and the irregular shape of less than 5 , respectively. After ball-milling at 800 rpm for 5 h, the powder mixture of Fe-based self-fluxing alloy powders and TiC particles formed into the agglomerated powders with the size of approximately 10 that was composed of the nanosized TiC particles and nano-sized alloy particles. The TiC particle-reinforced Fe-based self-fluxing alloy hybrid composite sintered at 1173 K revealed a much denser microstructure and higher micro-hardness than that sintered at 1073 K and 1273 K.

Mass production of graphene-based materials, which have high specific surface area, is of importance for industrial applications. Herein, we report on a facile approach to produce thermally modified graphene oxide (TMG) in large quantities. We performed this experiment with a hot plate under environments that have relatively low temperature and no using inert gas. TMG materials showed a high specific surface area (430 m2g-1). Successful reduction was confirmed by elemental analysis, X-ray photoelectron spectroscopy, thermogravimetic analysis, and X-ray diffraction. The resulting materials might be useful for various applications such as in rechargeable batteries, as hydrogen storage materials, as nano-fillers in composites, in ultracapacitors, and in chemical/bio sensors.

The experimental manipulation of protooncogenes and their gene products is a valuable research tool for the study of human neoplasia. In this study, the recently identified human cervical cancer protooncogene (HccR-2) was expressed in transgenic mice under the control of the tetracycline regulatory system. The phenotype observed was similar in many respects to human chronic neutrophilic leukemia (CNL). Thus, the HccR-2 transgenic mouse model is important not only for investigating the biological properties of the HccR-2 protooncogene in vivo, but also for analyzing the mechanisms involved in the progression of CNL.

Foraging preference in prey size and type is influenced by a variety of factors including energy requirements, season-dependent food availability, and social context (e.g. competition and predation risk). The oriental stork (Ciconia boyciana) is known as a wetland forager that inhabits human-managed wetlands such as paddy fields while breeding. However, it became an internationally endangered species. Information on its foraging preference is anticipated to play an important role in maintaining storks in captivity with a variety of food types as well as managing the food availability in foraging habitats of reintroduction sites. Specifically, the present study investigated the patterns of foraging preference of the subject in captivity as a partial study of the prerelease training and habitat management programs prior to reintroduction. The observations of foraging behavior of breeding adult storks included foraging preference in prey size (i.e. small, less than 6 cm, vs. large, larger than 6 cm, mudfish) during the incubation and nestling periods (March to April of 2009~2010) and prey type (i.e. mudfish, Misgurnus spp., crickets, Gryllus spp., and earthworms, Lumbricus spp.) during the postnesting period (October of 2009~2010). Our results indicated that storks in captivity not only preferred large to small mudfish independent of breeding stage but also preferred mudfish to crickets and earthworms. To our knowledge, captive storks did not appear to be constrained by providing offspring with various mudfish size and were likely dependent on mudfish, suggesting that a mudfish population in paddy fields should be monitored and managed for the main food resource for breeding storks prior to reintroduction.

The aim of this study was to investigate the suppressive activity of 70% ethanol extract and its dichloromethane fraction from Auricularia auricula-judae against adipogenesis and lipogenesis in 3T3-L1 preadipocytes. The ethanol extract and its dichloromethane fraction suppressed the differentiation and decreased lipid droplets in vitro. The dose-dependent increasing concentration of glycerol and lower triglycerides accumulation were found significantly (P < 0.05) with the treatment of both fractions from 70% ethanolic Auricularia auricula-judae extract and the glycerol-3-3phosphate dehydrogenase (GPDH) activity was also inhibited by both extracts. Further, the expression of adipogenic mRNAs were investigated by RT-PCR amplification. The key transcriptional factors, PPARγ and C/EBPα were decreased significantly at dose-dependent manner by both extracts of Auricularia auricula-judae. The expressions of LPL and FAS were also decreased by presence of these extracts. The decreased expressions of C/EBPβ, C/EBPγ and ACC1 were observed only by ethanol extract at 300 μg/ml concentration, while the expression of SREBP-1c, GLUT4 and aP2 were not altered in the 3T3-L1 adipocytes. These changes were occurred without the cytotoxic effect of both extracts against 3T3-L1 adipocytes in vitro. A positive control, fenofibrate inhibited the differentiation, triglycerides accumulation through PPARγ signaling by 2/3 reduction of PPARγ expression. Thus, these findings suggest that both extracts of Auricularia auricula-judae might be used to inhibit the differentiation of 3T3-L1 preadipocytes and reduction of triglycerides accumulation.

The aim of this study was to investigate the antitumor activity of solvent fractions from Auricularia auricula-judae 70% ethanol extract and confirmed the active components of dichloromethane fraction showing a potent antitumor activity than other fractions in the broncheoalveolar and gastric cancer cells. The solvent fractions of Auricularia auricula-judae extract, inhibited the growth proliferation of tumor cells in dose-dependent manner. The principle components of dichloromethane fraction were 5,11,17,23-tetrakis (1,1-dimethyl)-28-methoxypentacyclo [19.3.1.1 (3,7).1 (… (65.85%) and diazane (6.17%). The antitumor active components, diazane and gibberellic acid (GA3) were identified in this fraction by GC-MS analysis and lower antitumor activities than dichloromethane fraction. The unknown components of dichloromethane fraction were responsible for its cytotoxic effects on tumor cells. Based on IC50 value, gibberellic acid was little cytotoxic itself. According to PCR amplification, the apoptosis of tumor cells were induced by the down-regulation of Bcl-2 and over-expression of P53 on the presence of solvent fractions, diazane and gibberellic acid. Thus, these findings suggest that the dichloromethane might be used as functional feed additive that suppress the tumor growth in the body than other solvent fractions of Auricularia auricula-judae extracts.

Periodontitis results from the activation of host immune and inflammatory defense responses to subgingival plaque bacteria, most of which are gram-negative rods with lipopolysaccharides (LPSs) in their cell walls. LPSs have been known to induce proinflammatory responses and recently it was reported also that they induce the expression of microRNAs(miRNAs) in host cells. In our current study therefore, we aimed to examine and compare the miRNA expression patterns induced by the LPSs of major periodontopathogens in the human gingival epithelial cell line, Ca9-22. The cells were treated with 1 μg/ml of E. coli (Ec) LPS or 5 μg/ml of an LPS preparations from four periodontopathogens Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Aggregatibacter actinomycetemcomitans (Aa), and Fusobacterium nucleatum (Fn) for 24 h. After small RNA extraction from the treated cells, miRNA microarray analysis was carried out and characteristic expression profiles were observed. Fn LPS most actively induced miRNAs related to inflammation, followed by Aa LPS, Pi LPS, and Ec LPS. In contrast, Pg LPS only weakly activated miRNAs related to inflammation. Among the miRNAs induced by each LPS, miR-875-3p, miR-449b, and miR-520d-3p were found to be commonly up-regulated by all five LPS preparations, although at different levels. When we further compared the miRNA expression patterns induced by each LPS, Ec LPS and Pi LPS were the most similar although Fn LPS and Aa LPS also induced a similar miRNA expression pattern. In contrast, the miRNA profile induced by Pg LPS was quite distinctive compared with the other bacteria. In conclusion, miR-875- 3p, miR-449b, and miR-520d-3p miRNAs are potential targets for the diagnosis and treatment of periodontal inflammation induced by subgingival plaque biofilms. Furthermore, the observations in our current study provide new insights into the inflammatory miRNA response to periodontitis.

In this work, the effect of glycidyl methacrylate grafted multi-walled carbon nanotubes (GMA-MWCNTs) on the viscoelastic behaviors of polypropylene (PP) based nanocomposites was studied. The GMA-MWCNTs/PP was prepared using a bravender at 200℃ by melt mixing as a function of GMA-MWCNT content. The viscoelastic behaviors of GMA-MWCNTs/PP nanocomposites were measured by a rheometer. It was found that the GMA-MWCNTs were homogeneously dispersed in the PP matrix. The GMA-MWCNTs/PP nanocomposites showed higher storage modulus, loss modulus, and shear viscosity compared to pure PP nanocomposites and the maximum value was shown at 2.0 wt% GMA-MWCNTs loading. These results were probably attributed to the strong interfacial interaction between the GMA-MWCNT and the PP matrix.

In this study, the anti-oxidant, anti-tumorigenic, anti-hypertensive, anti-thrombic, anti-diabetic, and anti- inflammatory properties of 18 different species of genus Pleurotus were investigated. In addition, the amino acid, β-glucan, and polyphenol contents were also measured. The β-glucan and polyphenol contents were the highest in Pleurotus cornucopiae var. citrinopileatus (yellow Pleurotus) of all species. The yellow Pleurotus also exhibited the highest physiological activity as assessed by the DPPH IC, Cytotoxic activity, ACE activity and so on. To confirm the mechanism for the physiological activity of the yellow Pleurotus, we performed further examinations within ICR mice. The yellow Pleurotus reduced glucose, triglyceride and total cholesterol levels in the ICR mice blood for 4 weeks after feeding, and also significantly lowered both GOT and GPT levels. Taken together, our data indicates the yellow pleurotus is a promising functional food ingredient.

Mushroom is cultivated as one of the major economical crops in many areas of the Korea. The total production have steadily increased approximately 151,913 M/T in 2000 to 186,400 M/T in 2007. This study was carried out to investigate applicability of mushroom production using various organic media resources within the country. Eight organic resources were collected from various areas. Pleurotus ostreatus and Fulammulina velutipes showed the highest growth at the media of 10% red ginseng marc, 20% lacquer tree, 20% Juglans mandshurica, 10% Cudrania tricuspidata, 10~20% Acer pensylvanicum, 10% Lindera glauca. Mushroom mycelial growth at red ginseng marc media was slower than that of the control. But the sponin of the red ginseng was not detected at the fruiting body grown from red ginseng marc media, And three organic resources(barly powder, sweet potato powder, potato powder) was used to substitute rice bran used in mushroom cultivation. Pleurotus ostreatus and Fulammulina velutipes showed the highest growth at 10~30% sweet potato powder, 20% barly powder and 10% potato powder.

Auricularia auricula-judae has long been used as food and traditional remedies in Asian countries such as Korea and China. In this study, we evaluated the in vitro anti-tumor activity of various fractions from the ethanol extracts of Auricularia auricula-judae using various tumor cell lines. To do this, the mesh of Auricularia auricula-judae was mixed with 70% ethanol and heated at 1000C for 6 hrs and ethanol extract (ETOH) was collected. Ethanol extract was fractionated with dichloromethane (DCM), ethyl acetate, n-butanol and a water extract at room temperature as well as concentrated in a vacuum concentrator at a controlled temperature(<500C). The P388D1 macrophage and Sarcoma 180, human NSCLC NCI H358 (bronchioalveolar) and SNU1 cells (Gastric carcinoma) were cultured in RPMI. As the results, the cytotoxicity of the fractional extracts decreased significantly (P<0.05) in a dose-dependent manner. Dichloromethane extract (1 mg/ml) was the highest (P<0.05) in all experimental cell lines. There was also a significantly different sensitivity (P<0.05) among the P388D1, Sarcoma 180, NCI H358 and SNU1 cells for the fractional extracts. According to IC50 values, the most potent cytotoxic activity of dichloromethane fraction was found in Sarcoma 180 and NCI H358 cell lines. Butanol fraction appeared more cytotoxic to SNU1 cell line and water fraction had the highest cytotoxicity in P388D1 cell line. We did not find any significant difference between MTT and SRB assays in their ability to estimate cytotoxicity in all cell lines. Our findings suggest a potent antitumor activity of various fractions from the ethanol extracts of Auricularia auricula-judae depending on the solvent fractions and tumor cell lines. Further in vitro and in vivo studies will provide more information on the active compounds responsible for these activities and their potential as an anti-cancer remedy.