Embryonic stem (ES) cells are known to have an infinite proliferation and pluripotency that are associated with complex processes. The objective of this study was to examine expression of genes differentially regulated during differentiation of human ES cells by suppression subtractive hybridization (SSH). Human ES cells were induced to differentiate into neural precursor cells via embryoid body. Neural precursor cells were isolated physically based on morphological criteria. Immunocytochemical analysis showed expression of pax6 in neural precursor cells, confirming that the isolated cells were neural precursor cells. Undifferentiated human ES cells and neural precursor cells were subject to the SSH. TPX2 (Targeting Protein for Xklp2 (Xenopus centrosomal kinesin-like protein 2)) was identified, cloned and analyzed during differentiation of human ES cells into neural lineages. Expression of TPX2 was gradually down-regulated in embryoid bodies and neural precursor cells relative to undifferentiated ES cells. Targeting Protein for Xklp2 has been shown to be involved in cell division by interaction with microtubule development in cancer cells. Taken together, result of this study suggests that TPX2 may be involved in proliferation and differentiation of human ES cells.

The odorants from wastewater sludge treated with four different chemical oxidants, i. e., potassium ferrate, sodium hypochlorite, sodium permanganate, and calcium nitrate, were analyzed. The release of odorants from the treated sludge was not completely eliminated, only retarded, possibly due to the low one time doseof oxidants. In a comparison of the concentration profiles of methyl mercaptan and dimethyl sulfide, calcium nitrate was the best of the four different oxidants at reducing their emission. For methyl mercaptan, calcium nitrate gave the best result, while for dimethyl sulfide, potassium permanganate was found to be the best oxidant. From this study, it was found that the oxidation-reduction potential (ORP) would be an easy and inexpensive parameter for the monitoring of the release of offensive odors.

Hereditary dentin defects consists of dentin dysplasia(DD) and denti nogenesis imperfecta(Dr) ‘ The Dl associated with osteogenesis imperfecta has been classified as DI type 1. whereas isolated inherited defects have been categori zed as DI types II and III , However‘ whether DI type III should be considered a distinct phenotype 01' a variation of DI type 1I is debatable , Recent genetic findings have focused attention on the role of the dentin sialo phosphoprotein(DSPP) gene in the etiology of inherited defects of tooth dentin, We have identified novel mlltation( c,727G - > A, p,D243N) at the 243th codon of exon 4 of the DSPP gene in a Korean patient with DI type III The radiographic and histologic features of the patient revealed the classic phenotype of shell teeth These findings sllggest that DI type II and III are not separate diseases bllt rather the phenotypic variation 01' a s ingle disease

Dendropanax morbifera Léveille (Araliaceae) is an endemic species growing in the south-western part of South Korea and has been used in folk medicine and health functional food. Several studies have indicated that extract of D. morbifera (DP) has cytotoxic activities on a number of human cancers, such as, breast cancer, lung cancer, hepatoma, and chorioepithelioma. Recently, polyacetylene and triterpene compounds have been isolated from the DP and showed to have anti-complement activity. β-Amyrin, α-amyrin, dendropanoxide, and β-sitosterol are isolated from DP. However, its biological activities in cancer have not yet been clearly elucidated. In this study, we evaluated the anti-cancer activity of isolated triterpenoids from the DP leaves by measuring the levels of cytotoxicity against MCF-7 human breast cancer and A549 human lung cancer cells. Six triterpenoids were isolated from the n-hexane fraction of DP leaves along with the known compounds. β-amyrin (1), α-amyrin (2), olean-12-en-3,24 β-diol (3), dendropanoxide (4), β-sitosterol (5), and stigmasterol (6). Compound 3 and 6 were isolated from DP for the first time. Cytotoxic activities of six compounds were evaluated against two human cancer cell lines by using the MTT in vitro assay. Among them, five compounds (1, 2, 4, 5, and 6) showed moderate cytotoxic activities toward the tested cell lines, and were safe to normal cells. Western blot analysis showed a decreased expression of anti-apoptotic protein Bcl-2 and increased levels of pro-apoptotic protein Bax in MCF-7 and A549 cells treated by β-amyrin and α-amyrin. Flow cytometry analysis confirmed that five compounds (1, 2, 4, 5, and 6) treatment increased populations of sub-G1 (apoptosis) phase. The results of the present study revealed that triterpenoids from DP have the potential for further development as anticancer agents.

Interferon-stimulated gene 15 (ISG15) is known to interfere with viral replication and infection by limiting the viral infection of cells. Interferon-stimulated gene 15 (ISG15) interferes with viral replication and infectivity by limiting viral infection in cells. It also plays an important role in the immune response. In this study, tissue-specific expression of ISG15 in healthy rock bream samples and spatial and temporal expression analysis of rock bream ISG15 (RbISG15) were performed following rock bream iridovirus (RSIV) infection. RbISG15 expression was significantly higher in the eye, gill, intestine, kidney, liver, muscle, spleen, and stomach, but low in the brain. There were particularly high levels of expression in the liver and muscle. RbISG15 expression was also examined in several tissues and at various times following RSIV infection. ISG15 expression increased within 3 h in the whole body and decreased at 24 h after infection. In addition, temporal expression of several tissues following RSIV infection showed a similar pattern in the muscle, kidney, and spleen, increasing at 3 h and decreasing at 72 h. These results suggest that ISG15 plays an important role in the immune response of rock bream. Overall, this study characterizes the response of RbISG15 following RSIV infection.

Background: Mahonia Nepalensis DC. (Hoang lien o ro), the specie of the family Berberidaceae, is widely distributed in the high mountainous areas at altitudes 1700 – 1900 m of Vietnam. It is found that the stem of Mahonia nepalensis indicated anti-inflammatory, anti-bacterial and antifungal activities and they are used particularly for the treatment of eczema, psoriasis, and other skin conditions. However, no study on the antioxidant and anti-cancer activities of Mahonia Nepalensis stem has been previously reported. The aim of this study was to evaluate anti-oxidant and anti-cancer activities of Mahonia Nepalensis stem. Methods and Results: The stem pieces of Mahonia Nepalensis were dried and extracted three times with 100% methanol. After that, the extract was suspended in distilled water and then partitioned with n-hexane, ethyl-acetate (EtOAc) and butanol (water saturated BuOH) fractions were then evaporated using a vacuum rotary evaporator. Evaluation of the anti-oxidative activity of Mahonia Nepalensis was carried out using a DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-producing system. The results revealed that the ethyl acetate fraction of M. nepalensis possessed higher potential DPPH radical scavenging activity (IC50, 81.88 ± 1.33㎍/㎖) than other fractions as well as BHT (2,6-Di-tert-Butyl-4-methylphenol) (IC50, 250.49 ± 1.60㎍/㎖). The reducing power assay was also investigated and EtOAc fraction showed higher absorbance values than other fractions. At 1.0 mg/ml concentration, EtOAc fraction showed absorbance of 1.72, be higher than Ascorbic acid. Cell viability was evaluated according to the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyl tetrazolium Bromide) assay. By MTT assay, all fractions showed a significant reduction in cell viability on COLO 205 (Human colon carcinoma cell) at the highest concentration tested (200㎍/ ㎖) with over 70% decrease in cell viability was obtained, and the highest significantly inhibiting effect occurred in butanol fraction with approximately 90% reduction in cell viability. Conclusion: We demonstrated that Mahonia Nepalensis stem extract has highly potential in anti-cancer activity. Further studies are necessary in order to explore the variety of Mahonia Nepalensis stem to be applied as a valuable natural material.

Background : Sea buckthorn (Hippophae rhamnoides L.) was used as medicinal plant in Tibetan and Mongolian traditional medicines. It has been recognised as a versatile nutraceutical crop with diverse uses for the treatment of diseases, such as gastric ulcers, lung disorders, cardiovascular diseases, mucosal injuries and skin disorders. Physiological research on mixture of sea buckthorn leaf and fruit have not be reported. Therefore, in this study, using sea buckthorn mixture, antioxidant and anti-inflammatory effects were determined. Methods and Results : The experiment was carried out using 11 samples (100% leaf extract - 100% fruit juice powder). The antioxidant and free radical scavenging activities of sea buckthorn mixture were evaluated using DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. The leaf extract with fruit juice in the ratio of 60 : 40 (w/w) showed a significant effect (86.43%). The mixture of sea buckthorn leaf and fruit were investigated for anti-inflammatory activity using LPS stimulated Raw 264.7 cells. The results showed that the higher ratio of leaf extract indicated greater anti-inflammatory activity (approximately 10%, NO production ). Conclusion : These result showed that the mixture of sea buckthorn leaf and fruit can be used as a variety of antioxidant and other functional product research and development processes as valuable natural materials.

Lectins belong to the pattern-recognition receptors (PRRs) class and play important roles in the recognition and elimination of pathogens via the innate immune system. Recently, it was reported that lily-type lectin-1 is involved when a pathogen attacks in the early immune response of fish. However, this study is limited to information that the lectin is involved in the innate immune response against viral infection. In the present study, the lily-type lectin-2 and -3 of Oplegnathus fasciatus (OfLTL-2 and 3) have been presented to be included B-lectin domain and two D-mannose binding sites in the amino acid sequence that an important feature for the fundamental structure. To investigate the functional properties of OfLTLs, the tissue distribution in the healthy rock bream and temporal expression during early developmental stage analysis are performed using quantitative real-time PCR. OfLTL-2 and 3 are predominantly expressed in the liver and skin, but rarely expressed in other organ. Also, the transcripts of OfLTLs are not expressed during the early developmental stage but its transcripts are increased after immune-related organs which are fully formed. In the challenge experiment with RBIV (rock bream iridovirus), the expression of OfLTLs was increased much more strongly in the late response than the early, unlike previously known. These results suggest that OfLTLs are specifically expressed in the immune-related tissues when those organs are fully formed and it can be inferred that the more intensively involved in the second half to the virus infection.

Background : Haskap berries commonly refer to fruits of Lonicera caerulea L., recognized by the Japanese aborigines as the “The elixir of life.”. Due to their recent arrival on the North American market, haskap berries have not yet been positioned among other berries and compared in terms of their phytochemical content. And haskap berries have higher ascorbic acid and anthocyanin content than other berries known for their health-promoting benefits, such as blueberries. However, no study has reported on the antioxidant and anti-cancer activity of Lonicera caerulea stem. The purpose of this study is to present the current research on the chemical content, antioxidant and anti-cancer activities of Lonicera caerulea stem. Methods and Results : The stem of Lonicera caerulea L. ware dried in the shade at room temperature and extracted with 100% methanol. The extract was suspended in deionized water and partitioned sequentially with n-hexane, chloroform, ethyl-acetate and butanol (water saturated BuOH) fractions. Antioxidant activities were measured by determination of antioxidants, DPPH (2,2-diphenyl-1-picrylhydrazyl). Cell viability was determined by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. All cell lines were purchased from the Korean Cell Line Bank (Seoul, Korea). All results were performed with three replications were processed statistically. By DPPH assay, the Lonicera caerulea L. the highest activity was obtained from the ethyl-acetate fraction (IC50=15.46 ㎍/㎖). By MTT assay, the chloroform fraction showed a significant growth inhibiting effect on MCF-7 (Human breast cancer, IC50=225.91 ㎍/㎖), COLO 205 (Human colon cancer, IC50=179.55 ㎍/㎖), but on AGS (Human stomach cancer) and other fractions it did not show effect. Conclusion : We demonstrated that Lonicera caerulea L. stem extract and fractions has antioxidant and antiproliferation activity in vitro. Further studies should identify the active constituents in Lonicera caerulea L stem to evaluate the potential in vitro antioxidant and antiproliferation activities of the extract.

Background : Astilboides tabularis (Hemsl.) Engl. is a perennial herbaceous plant, distributed in the northern high mountains of the Korean peninsula and China. It is an excellent ornamental plant currently at risk of overharvesting and therefore, is designated as an endangered wild plant Class II by the Ministry of Environment. Physiological research on A. tabularis has not be reported. Therefore, in this study, using A. tabulari extracts, antioxidant and Anti-inflammatory effects were determined. Methods and Results : The antioxidant and free radical scavenging activities of A. tabularis extracts were evaluated using DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. The results showed that the ethyl acetate fraction of A. tabularis possesses potent DPPH radical scavenging activity (2.90±0.08㎍/㎖), similar to the scavenging activity of ascorbic acid (2.19±0.06㎍/㎖), and better than the powerful antioxidant α-tocopherol (10.60±0.40㎍/㎖) as well as BHA (butylatedhydroxy anisole)(6.12±0.27㎍/㎖). The ethyl acetate fraction possessed a significantly higher concentration of total phenolic (549.70±2.72㎎GAE/g) and flavonolic content (154.58±1.04㎎QE/g). It was also found that the ethyl acetate fraction exhibited high reducing power and inhibition of ROS (Reactive Oxygen Species) formation. Different fractions of A. tabularis were tested for anti-inflammatory activity using LPS stimulated Raw 264.7 cells. The n-hexane and ethyl acetate fractions exhibited a high inhibitory effect on NO (Nitrite oxide) production (22.43±1.06%, 19.30±0.45%, respectively) at 200㎍/㎖ concentration. The mRNA of IL-1β, iNOS and COX-2 gene expression was decreased by treatment with the ethyl acetate fraction. These results showed that A. tabularis extracts can be used as natural substances to control inflammation. Conclusion : These result showed that A. tabularis extracts can be used in a variety of antioxidant and other functional product research and development processes as valuable natural materials.

Background : This study aimed to investigate the antioxidant and anti-inflammatory activities of water chestnut (Trapa japonica Flerow) extract. Methods and Results : The antioxidant and anti-inflammatory activities of 100% methanol extract of water chestnut were investigated. The methanol extract was evaluated for its total phenolic and flavonoid content, DPPH•(1,1-diphenyl-2-picrylhydrazyl) free-radical scavenging activity,reducing power, andeffect on nitric oxide (NO) production and cell viability using real-time polymerase chain reaction (qPCR). The total phenolic content was 438.31 ㎍ allic acid equivalent (GAE)/㎎ extract and the total flavonoid content was 61.40 ㎍ quercetin equivalent (QE)/㎎ extract. In addition, results revealed the extract possessed antioxidant activity (DPPH• free-radical scavenging activity) with IC50 value of 5.28 ㎍㎖ The reducing power of the extract was assayed spectro photometrically and showed Abs of 0.71 at 100 ㎍㎖ Furthermore, extracts of water chestnut exhibited no cytotoxicity in RAW 264.7 cells. In addition, the NO assay revealed that LPS-induced NO production was significantly inhibited following treatment with water chestnut extracts. The expression of pro-inflammatory proteins such as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 decreased in a concentration-dependent manner. The water chestnut extract also decreased tumor necrosis factor α (TNF-α) release. Conclusion : Therefore, the present findings provide scientific evidence for the nutritional potential, chemical composition, and biological activities of Trapa japonica Flerow anddemonstrate its potential use as a functional food forapplication in the pharmaceutical industry

The purpose of this study was to develop a supplemental healthy food that can help prevent high blood pressure-related diseases caused due to the excessive consumption of sodium in salt. This was achieved by using ion-displacement techniques to produce mineral salt with lower sodium content by using fermented brown rice by-products rich in minerals. Mineral salt containing 2019.2 mg/100 g of potassium, 678.5 mg/100 g of magnesium, 48.7 mg/100 g of calcium, and 19.5 mg/100 g of sodium was obtained by fermenting brown rice by-products to create a culture medium for the mineral salt. Mineral salt containing 1769.7 mg/100 g of potassium, 573.6 mg/100 g of magnesium, 35.3 mg/100 g of calcium, and 19.5 mg/100 g of sodium was obtained by filtering and refining the by-product extract of fermented brown rice. The results showed that when the stream velocity of the instrument used for electrolysis was 200 mL/min and the current and the concentration of the reactive liquid in the purified water chamber were higher, the effect of electrolysis was greater. Ion hot water extraction of the fermented brown rice by-products improved by up to 95% and was collected as purified water within 90 min of the reaction time. Chloride ions with pH 7.4 were produced by mixing sodium hydroxide in a purified saline water chamber with electro-analyzed water. The salt produced in this study contained low sodium, 5.7~30%, as compared to 40% sodium content of the normal salt.

The objective of the study was to identify 52 Asian pear accessions, two primary pear species, and one reference pear Asian pear with 12 microsatellite markers to maintain pear germplasm collection. The number of alleles of 12 microsatellites detected ranged from eight at CH03d12 to 18 at CH01f07. Gene diversity ranged from 0.7053 at CH01d08 to 0.9224 at CH01f07. The lowest value of PIC was 0.6600 at CH01d08 and the highest was 0.9171 at CH01f07. A group consisting of ‘Ooharabeni,’ ‘Bartlett,’ and P. calleryana was out-grouped and served as a reference to determine the relationship among Asian pear accessions. Except for the out-group, 50 Asian pears were segregated into two groups. Group I was divided in two small groups. Each small group was characterized by P. bretschneideri and P. ussuriensis, respectively. Group II was characterized as P. pyrifolia, and the group was divided in four small groups. The eigenvalue, difference, proportion, and cumulative of six principal components based on PCA to 12 microsatellite. The eigenvalue of the first principal components was 5.5850. The proportion of the first principal component was 0.9308. The cumulative value of the first two principal components was 0.9801. Consequently, nearly all of the results were elucidated by the two principal components. The results from analysis of the standard set of microsatellites in this study may be used as basic materials for the management of Asian pear germplasm collections, and the data might be useful in the development of a core collection.

Proline has been shown to accumulate in plant under various type of stresses. In our previous study, changes in cold hardiness and proline content showed contrasting patterns during a constant deacclimation. This study was performed to investigate the proline accumulation and related gene expression in response to repeated deacclimation and reacclimation in peach cultivar ‘Daewol’. Proline content was analyzed using the ninhydrin method and related gene expressions were examined using quantitative real-time RT-PCR. Proline contents of ‘Daewol’ increased during the repeated deacclimation treatments. Interestingly, during the twice deacclimation, expressions of P5CS (Δ1-pyrroline-5-carboxylatesynthase) constantly decreased, whereas expressions of P5CR (Δ1-pyrroline-5-carboxylatereductase) increased. Expressions of OAT (ornithine-δ-aminotransferase) indicated up- and down- pattern in response to repeated deacclimation and reacclimation. Our results indicated that proline responds positively to higher temperature in the shoots of peach cultivar ‘Daewol’ and expressions of both P5CS and P5CR genes could show contrasting patterns during the deacclimation. Moreover, our results suggest that ornithine pathway could serve as an alternative pathway in proline synthesis process during deacclimation in peach.

Proline (Pro) accumulation is a common physiological reaction in response to abiotic stresses in many plants. Accumulation of Pro is believed to play the important role in protecting cellular components from dehydrating effects due to such stresses. The study was performed to investigate the relationship between cold hardiness and Pro content or expression of related genes in peach cultivars during a constant experimental deacclimation. Changes in cold hardiness were determined using electrolyte leakage method in the shoots of 10 peach cultivars (Prunus persica ‘Aikawanakajima’, ‘Chiyomaru’, ‘Daewol’, ‘Janghowon Hwangdo’, ‘Kiraranokiwami’, ‘Mihong’, ‘Misshong’, ‘Soomee’, ‘Suhong’, and ‘Sun Gold’). Pro content was analyzed using the ninhydrin method and related gene expressions were examined using quantitative real-time RT-PCR. While cold hardiness of 10 peach cultivars decreased, Pro contents of those increased during the deacclimation. Notably, at the same time, expression of P5CS (Δ1-pyrroline-5-carboxylatesynthase) decreased in 10 peach cultivars, whereas expressions of P5CR (Δ1-pyrroline-5-carboxylatereductase) and OAT (ornithine-δ-aminotransferase) increased. Our results demonstrate that Pro responds positively to higher temperature in the shoots of 10 peach cultivars and expression of both P5CS and P5CR genes could show contrasting patterns during the deacclimation. Furthermore, our results suggest that ornithine pathway, which has been suggested to be important during seedling development, could serve as an alternative pathway in Pro synthesis process during the deacclimation in peach.

In this study, we sought to identify primary pears species and Korean native pears, without the use of morphological characteristics. In addition, this study was to establish pear DNA fingerprinting data for Korean native pears using 12 microsatellite markers, and to accurately classify a database for management of the Korean pear collection. Forty two pear accessions (7 primary pears, 5 Asian pears, 29 Korean pears, and 2 reference pears) were analyzed with twelve primers covering whole pear genome. In the present study, all pear accessions were successfully classified along with their pedigrees, and the distribution of primary pears was parallel to those of the previous taxonomic results. Korean pears were divided into 3 groups. Group I was characterized by Pyrus calleryana, and included Korean pea pears. Group II was characterized by P. pyrifolia, and was classified into 2 small groups. The first small group comprised of ‘Najucheongbae’, ‘Sunchanggulimdolbae’, ‘Andongmookbae’, ‘Andongdangsilri’, and ‘Najucheongbae’ and was presumed to be cultivars of P. pyrifolia. The second small group consisted of ‘Cheongdangrori’ and ‘Pyeongchangsuhyangri’. These two accessions were assumed to be a hybrid of P. pyrifolia and the other cultivar. Group III was characterized by P. ussuriensis. ‘Goesanhwangbae’, ‘Andongcheongsilri’, ‘Gongjucheongsilri’, and ‘Yecheoncheongbae’ were assumed to be cultivars of P. ussuriensis. Contrary to ‘Ulreungdocheongbae A’, ‘Ulreungdocheongbae B’ was classified as belonging to the P. ussuriensis group. It is possible that this is a consequence of, P. ussuriensis genes being transferred into ‘Ulreungdocheongbae B’. The result of this research reaffirmed the efficiency of a standard set of microsatellite markers and provides data, which will be useful for developing a core collection of pears.

To successful molecular breeding, identification and functional characterization of breeding related genes and development of molecular breeding techniques using DNA markers are essential. Although the development of a useful marker is difficult in the aspect of time, cost and effort, many markers are being developed to be used in molecular breeding and developed markers have been used in many fields. Single nucleotide polymorphisms (SNPs) markers were widely used for genomic research and breeding, but has hardly been validated for screening functional genes in olive flounder. We identified single nucleotide polymorphisms (SNPs) from expressed sequence tag (EST) database in olive flounder; out of a total 4,327 ESTs, 693 contigs and 514 SNPs were detected in total EST, and these substitutions include 297 transitions and 217 transversions. As a result, 144 SNP markers were developed on the basis of 514 SNP to selection of useful gene region, and then applied to each of eight wild and culture olive flounder (total 16 samples). In our experimental result, only 32 markers had detected polymorphism in sample, also identified 21 transitions and 11 transversions, whereas indel was not detected in polymorphic SNPs. Heterozygosity of wild and cultured olive flounder using the 32 SNP markers is 0.34 and 0.29, respectively. In conclusion, we identified SNP and polymorphism in olive flounder using newly designed marker, it supports that developed markers are suitable for SNP detection and diversity analysis in olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.