Astragalus membranaceus is a well known oriental medicinal herb. The polysaccharides of the aboveground parts (AMA) and the radix (AMR) of A. membranaceus are the most important functional constituents. Methods and Results: The aim of this study was to determine the effects of AMA and AMR on the oxidative damage induced in the skeletal muscle of rats subjected to exhaustive exercise. Sprague-Dawley rats were randomly divided into exercise and non-exercise groups; in the groups receiving the test compounds, AMA and AMR were administered orally for 30 days. Skeletal muscle samples were collected from each rat after running to exhaustion on a treadmill to determine the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) and the concentation of malondialdehyde (MDA). The antioxidant enzyme activities of SOD and GSH-Px of skeletal muscle of AMA- and AMR-treated groups were significantly higher than those of the control and commercial sports drink (SPD)-treated groups in exhaustive exercise rats. In addition, MDA concentrations in the skeletal muscle of the AMA- and AMR-treated groups were significantly lower than those of the control and SPD-treated groups. In the present study, the effects of AMA and AMR on exercise endurance capacity were also evaluated in mice subjected to a swimming exercise test. AMA and AMR supplementation prolonged the swimming time of mice and enhanced exercise endurance capacity. AMA and AMR possess the ability to retard and lower the production of blood lactate, and prevent the decrease of serum blood glucose. Conclusions: These results showed that, AMR and AMA exerted beneficial effect in mice, increasing the activity of the antioxidant systems and inhibiting oxidative stress induced by exhaustive exercise. The compounds improved exercise performance and showed anti-fatigue effects against exhaustive exercise.

Background: This study were to investigate the effect of Pediococcus pentosaceus fermented Radix astragali (AMRP) and non-fermented products (AMRNP) on collagen synthesis in the cultures of human dermal fibroblasts, and their inhibitory effects on the matrix-degrading enzymes (collagenase, elastase, and gelatinase). Methods and Results: Both AMRP and AMRNP significantly improved cell growth and proliferation of HDF cells. However, the enzyme-linked immunosorbent assay and Western blot analysis demonstrated that AMRP, but not AMRNP, significantly and dose-dependently stimulated the biosynthesis of type I procollagen in both aged (74 y) and young (21 y) HDF cells. Real-time reverse transcription-polymerase chain reaction revealed that expression of type I, type III procollagen and transforming growth factor β1 (TGF-β1) mRNA was significantly stronger in AMRP-treated HDF cells than that of AMRNP-treated and un-treated HDF cells. The AMRP revealed an increase in astragaloside Ⅳ only depending on increase in fermentation period, because other astragalside converted to astragaloside Ⅳ, which it detached acyl group by fermentation processing of Pediococcus pentosaceus. Conclusion: The results also suggested that AMRP could stimulate the collagen biosynthesis in human dermal fibroblasts, which is, associated with the regulation of procollagen biosynthesis resulting from AMRP-induced TGF-β1 expression and the mitogenic activity in HDF cells, and therefore, is expected to reduce the age-dependent loss of extracellular matrix proteins.

Background: Hot steaming is known to be effective in improving the biological activities of plant extracts by breaking down useful compounds to low molecular weight ones. Methods and Results: This study aimed to develop an optimal extraction and steam processing method for enhancing the low molecular ginsenoside contents of the adventitious roots culture of wild mountain ginseng. The total ginsenoside was optimally extracted when 70% EtOH was used at 50℃, whereas low molecule ginsenoside such as Rg2, Rh1, Rh4 and Rk1 could be extracted using 70% EtOH at 70℃. The adventitious roots culture of wild mountain ginseng is known to contain four major ginsenosides, i.e., Rb2, Rb1, Rg1 and Rd, however new ginsenosides Rg6, Rh4, Rg3, Rk1 and Rg5 were new abundantly obtaind after steam processing method was applied. The contents of total ginsenosides were the highest when thermal steam processing was conducted at 120℃ for 120 min. Unlike ginsenosides such as Rg1, Re, Rb1, Rc, Rb2, and Rh1, which decreased after steam processing, Rg3, Rk1, and Rg5 increased after thermal processing. Steam processing significanltly reduced the content of Rb1, increased that of Rg6 by about ten times than that in the adventitious roots culture of wild mountain ginseng. Conclusions: Our study showed that the optimal extraction and steam processing method increased the content of total ginsenosides and allowed the extraction of minor ginsenosides from major ones.

Background: Polysaccharides are the most important functional constituent in Astragalus membranaceus. The purpose of the present study was to evaluate the effect of polysaccharides isolated from the aboveground parts of A. membranaceus (AMA) and polysaccharides isolated from the roots of A. membranaceus (AMR) immune function by modulated cytotoxic T cell and Th1- and Th2- related cytokines kinetics. Methods and Results: Sprague-Dawley rats were randomly divided into exhaustive exercise case groups and non-exercise case, AMA and AMR samples were administered orally for 30 days (500 ㎎/㎏/day and 10 ㎎/㎏/day, respectively) and were compared to those rats in the groups fed commercial sports drink (SPD) and vehicle. Both exhaustive exercise groups and non-exercise groups had a lower ratio of CD4+ and CD8+ cells in the spleens of the rat fed AMA and AMR compared to those in the rats fed SPD and vehicle group. These results suggested that AMA and AMR promote an increase in the proportion of cytotoxic T cells. The IL-4- producing T lymphocytes decreased significantly in the AMR (10 ㎎/㎏/day) group compared to SPD and vehicle, whereas the AMA group increased the IL-4 concentration more than the SPD and vehicle in exhaustive exercise group. However, the populations of IFN-γ-producing T lymphocytes of AMR and AMA increased. AMA decreased the concentration of IFN-γ to inhibit the Th1 response and thereby increased the concentration of IL-4 to induce a Th2 response that was related to humoral immunity in the non-exercise group. Conclusions: These results showed that, in addition to Th1/Th2 regulation, AMR and AMA played an important immuno-modulatory role after exhaustive exercise-induced Th1/Th2 lymphocyte imbalance, which might be correlated with cytokine producing immunoregulatory cells.

Backgrounds : The action mechanisms of several chemopreventive agents derived from herbal medicine and edible plants have become attractive issues in cancer research. Tea is the most widely consumed beverage worldwide. It have been demonstrated that the active principles of tea sources such as flower extract Camellia sinensis (CSF) and Camellia japonica (CJF)were attributed to their tea polyphenols. We focused on investigating CSF, CJF, mixtures of CSF and CJF has been proven to suppress colonic tumorigenesis. Methods and Results : In this study, human colorectal carcinoma HT-29 cells were treated with CSF, CJF, mixture of CSF and CJF to examine the anti-proliferative and pro-apoptotic effects of mixture of CSF and CJF (3 : 1), as well as the molecular mechanism underlying these effects. Cell viability assay, nuclear staining, DNA fragmentation, caspase assay, cytochrome c release, were utilized to dissect the signaling pathways. In mixture of CSF and CJF (3 : 1), CSF appeared most anticancer effect by both MTT assays and the cleavage analysis of apoptosis-related molecules and PARP. Interestingly, we found that CJF make it possible to express the apotosis inducing by CSF in a short time and apoptosis effect of CSF maintained sustainable. Conclusion : In summary, our results from this study suggest that in HT-29 human colon cancer cells (i) CSF treatment causes damage to mitochondria, and (ii) CJF contributed CSF induced apoptotic cell death mediates cytochrome C release, (ⅲ) mixture of CSF and CJF (3 : 1) the potential to function as a chemopreventive agent against colon cancer.

Backgrounds : Camellia sinensis is known to have a very high antioxidant activity, but its petals are small and it is difficult to use it because of its low yield. In comparison Camellia japonica has many petals and yield, however, the biological effects of C. japonica have been less frequently studied. In the present study, we focused on investigating the in vitro antioxidant effect of the ethanol extract from flower of C. sinensis (CSF), C. japonica (CJF) and mixture of CSF and CJF. Methods and Results : Content of total phenolics and total flavonid, DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activities, reducing power, superoxide anion and hydroxy radical scavenging activity of CSF and CJF were compared in vitro experimetal models. Total phenolics was contained the higer in CJF (172.19±1.65 mgCAE/gEX) than 146.75±0.15 mgCAE/gEX in CSF. And effects of antioxidant measured by reducing power, superoxide anion generated by xanthine/xanthine oxidase and hydroxyl radical generated by the Fenton reaction (FeSO4 + H2O2) in a cell-free system was shown higher in mixture of CSF and CJF than BHT, ascorbic acid as a chemical oxidant which was detected by electron spin resonance spectrometry coupled with DMPO spin trapping. These results suggest that Camellia flower extract such as CSF and CJF exhibits antioxidant properties by scavenging ROS. Camellia extract contained quercetin, quercetin-3-O-glucoside, quercitrin and kaempferol, which are antioxidant compounds. Conclusion : As a result, the combination of CSF and CJF showed higher antioxidative effect than using CSF or CJF alone.

Abeliophyllum distichum is a monotypic taxon of Oleaceae and endemic to Korea. A comprehensive study on embryogeny and fruit and seed coat ontogeny in Abeliophyllum was carried out via microtome and light microscopy. The fertilization occurs during mid– to late April and embryo matures by early July. The embryo development follows the general fashion from globular embryo – transition embryo – heart shaped embryo – torpedo embryo – walking-stick embryo to mature embryo. The pericarp clearly differentiates into three histological zones: exocarp, mesocarp, and endocarp. The young seed comprises 10-12 cells thick seed coat and the mature seed coat comprises an exotesta, 6-8 mesotesta and an endotesta. Any crystals, phenolic-like compounds, idioblasts, and the sclereids are not found in pericarp as well as seed coat. An overall development confirms Solanade type of embryogenesis in Abeliophyllum. The endocarp becomes more prominent in mature fruit

The THO/TREX complex mediates the transport of nascent mRNAs from the nucleus towards the cytoplasm in animals, and it has a role in small RNA-dependent processes in plants. Here we describe five mutant alleles of Arabidopsis thaliana THO2, whichencodes a core subunit of the plant THO/TREX complex. tho2 mutants present strong developmental defects resembling those in plants compromised in microRNA (miRNA) activity. In agreement, not only the levels of siRNAs, but also of mature miRNAs were reduced in tho2 mutants. As a consequence miRNA target mRNAs accumulated to higher levels than in wild type. Yeast two hybrid experiments showed that THO2 does not seem to interact with any of the known miRNA biogenesis components, implying a more indirect role of THOs in small RNA biogenesis. We also detected alterations in the splicing pattern of genes encoding Serine/Arginine-rich proteins in tho2 mutants, suggesting a previously unappreciated role of the THO/TREX complex in alternative splicing.

VitE (tocotrienols and tocopherols) are micronutrients with antioxidant properties synthesized by photosynthetic bacteria and plants that play important roles in animal and human nutrition. A new mutant line, T1001-1, was isolated from in vitro mutagenized population by ionizing radiation and shown to have increased VitE contents. The total VitE content was 26% increased in the T1001-1 mutant seeds compare with cv. Dongan (wild-type). In addition, we showed that the mutant confers retarded seedling growth during the early seedling growth stage in rice. To study the molecular mechanism of VitE biosynthesis, we used the rice microarray to identify genes that are upor down-regulated in T1001-1 mutant. In addition, we identified differentially regulated pathway using MapMan analysis, which provides deep insight into changes in transcript and metabolites. Our results enhanced the transcription of genes involved in starch and lipid metabolism in T1001-1 mutant. To identify the molecular mechanisms of the events involving transcription factors in tocopherol accumulation, we compared the expression patterns of transcription factors. The AP2-EREBP, WRKY, C2H2 transcription factor were up-regulated, whereas the MYB family was down-regulated in T1001-1 mutant. Our results demonstrate change of important transcript in high level of VitE accumulating rice mutant.

Manganese () is a trace element that is essential for normal physiology, and is predominantly obtained from food. Several lines of evidence, however, demonstrated that overexposure to exerts serious neurotoxicity, immunotoxicity and developmental toxicity, particularly in male. The present study aimed to evaluate the effect of 0, 1.0, 3.3, and 10 mg/kg/day doses of on the reproductive organs in the immature female rats. Rats (PND 22; S.D. strain) were exposed to () dissolved in drinking water for 2 weeks. The animals were sacrificed on PND 35, then the tissues were immediately removed and weighed. Histological studies were performed using the uteri tissue samples. Serum LH and FSH levels were measured with the specific ELISA kits. Body weights of the experimental group animals were not significantly different from those of control group animals. However, ovarian tissue weights in 1 mg and 3.3 mg dose groups were significantly lower than those of control animals (p<0.05 and p<0.01, respectively). Uterine tissue weights of 3.3 mg dose groups were significantly lower than those of control animals (p<0.01), while the 1 mg dose and 10 mg dose failed to induce any change in uterine weight. Similarly, only 3.3 mg dose could induce the significant decrease in the oviduct weight compared to the control group (p<0.05). Non-reproductive tissues such as adrenal and kidney failed to respond to all doses of exposure. The uterine histology revealed that the exposure could affect the myometrial cell proliferation particularly in 3.3 mg dose and 10mg dose group. Serum FSH levels were significantly decreased in 1mg dose and 10 mg groups (p<0.05 and p<0.01, respectively). In contrast, treatment with 1 mg dose induced a significant increment of serum LH level (p<0.05). The present study demonstrated that exposure is capable of inducing abnormal development of reproductive tissues, at least to some extent, and altered gonadotropin secretions in immature female rats. Combined with the well-defined actions of this metal on GnRH and prolactin secretion, one can suggest the might be a potential environmental mediator which is involved in the female pubertal process.

Molecular genetic markers have been widely used as powerful tools for analyzing the genome. In particular, SSR markers have practical applications in breeding systems because they can be used in high-throughput analyses for genetic mapping, heritable diversity testing, purity analysis, and marker-assisted selection. Currently, due to technical advances in DNA sequencing, large sequence databases are available for large-scale SSR mining and marker development. Here, we describe an automated method, the SSR Finder program, for SSR discovery in the onion sequence database, and primer design for amplifying the detected SSRs. A total of 1,049 SSR primers were obtained for genetic purity testing, and 100 SSR sets were analyzed in 14 bulb onion breeding lines using clustering analysis. A total of 20 selected markers from screening of all 1,049 SSR primers, were finally applied for genetic purity testing in three breeding lines, NW1, NW9, and NW10. The initial tests showed that 15%, 71%, and 97% of individuals within NW1, NW9, and NW10, respectively, were genetically homogeneous. These markers produced using the SSR Finder will be useful for investigating the genetic purity of onion breeding lines.

Ionizing radiation is known to cause chromosomal alterations such as inversions and deletions and affects gene expression within the plant genome. To monitor the genome-wide transcriptome changes by ionizing radiation, we used rice Affimetrix GeneChip microarray to identify genes that are up- or down regulated by gamma-ray (200 Gy, 60Co source), cosmic-ray and ion beam (40 Gy, 220 MeV carbon ion). The overall expression patterns between gamma-ray and ion beam were similar but cosmic-ray was regulated differently. Combined results from all 3 radiations identified 27 up-regulated genes and 188 down regulated genes. These results mean the induction of similar mechanism changes in treatments of gamma ray and ion beam. However the different expression in treatment of cosmic-ray might be due to the other environmental conditions. Among the commonly up- or down- regulated genes, we chose highly up- or down- regulated several genes and confirmed its regulation in response to ionizing radiation exposure by RT-PCR analysis. Moreover, we showed that specific co-expression networks of candidate radio marker genes by ARACNE algorithm. Our results present profiles of gene expression related to different ionizing radiation and marker gene to predict sensitivity to ionizing radiation, such as GS (glutelin subunit) and FBX322.

The canine major histocompatibility complex (MHC) is referred to dog leukocyte antigens (DLA), which is known to be the most polymorphic genetic system in canine species. Many cloned dogs have been produced since Snuppy, first cloned dog, there was no research about genetic identity of MHC among cloned animals. Recently in Lee’s group, two non-transgenic cloned beagles (BG1, 2) were produced by somatic cell nuclear transfer (SCNT) using fetal fibroblast (BF). Also, four transgenic cloned beagles (Ruppy 1-3, 5) were generated using transgenic BF transfected with Red fluorescent protein (RFP) gene. We hypothesize that non-transgenic (BG1, 2) and transgenic (Ruppy 1-3, 5) cloned beagles derived from identical donor cells have the same immunological genetic characteristic except for RFP gene insertion in the genome. Thus, the aim of this study is to confirm the immunological identity of DLA class II in cloned beagles produced using same nuclear donor cell. Genomic DNA was extracted from blood of BG1, BG2, Ruppy 1, 2, 3 and 5. Genomic DNA of normal two control beagle, no correlation with BF was also investigated for rulling out the possibility that beagles were inbred. Forward and reverse primers used for DLA-DQA1 and DQB1 respectively were DQAF: 5’-TAAGGTTCTTTTCTCCCTCT-3’ and DQAR: 5’-GGACAGATTCAGTGAAGAGA-3’ DQBR:5’-CTCACTGGCCCGGCTGTCTC-3’ and DQBR: 5’-CACCTCGC CGCTGCAACGTG-3’. Polymerase Chain Reaction (PCR) products were purified, sequenced directly using the Big Dye Terminator kit. Sequencing analysis was performed on an automated 3730xl DNA analyzer. In experiment 1, sequence of DLA-DQ alpha 1 (DQA1) and DLA-DQ beta 1 (DQB1) exon 2, hypervariabel region, was compared in BG1 and BG2. Experiment 2 also compared the sequence of DQA1 and DQB1 among Ruppy 1, 2, 3 and 5. Experimental 3 compared sequence of DQA1 and DQB1 among all cloned dogs (BG1, BG2 and Ruppy 1-3, 5). As a result, BG1 and BG2 have same allele for DQA1 and DQB1 as we expected. They share DQA1*00101 and DQB1*02901 in experiment 1. In experiment 2, Ruppy 1, 2, 3 and 5 also have identical DQA1*00101 and DQB1*02901 allele. No discrimination between transgenic dogs and cloned dogs was seen in DQA1 and DQB1 Allele in experiment 3. DQA1, DQB1 allele was identified as *00101 and *02901 in all dogs. We provided the allele identity of DQA1and DQB1 in cloned beagles, which can be used as preliminary data for immunological related studies. In conclusion, transgenic cloned dogs despite of red fluorescent protein genes being inserted in their nuclear DNA were immunologically compatible with non-transgenic cloned dogs. We demonstrated that cloned beagles produced using identical nuclear donor were immunologically compatible.

A resveratrol synthase (RS) gene was isolated from peanut (Arachis hypogaea, L. cv. Jinpoong) plants. This gene was placed under the control of the cauliflower mosaic virus 35S promoter (CaMV35S) and introduced into two Korean varieties of potato (Solanum tuberosum L. cvs. Jasim and Jowon) plants by Agrobacterium-mediated gene transfer. Putative transformants were screened by PCR with primers designed from CaMV 35S promoter, NOS terminator and RS gene. Most of selected transgenic potato plants showed the amplification of expected fragments by PCR of genomic DNA with gene-specific primers, while they were absent in untransformed control plants. Expression of the resveratrol synthase gene was also examined by northern blot analysis. The transformants showed a band which was lacking in the control plant, confirming that the introduced gene is transcribed into mRNA in the transformants. The strength of the band, which reflected the level of mRNA expression, differed among the individual transformants. Among the transformants obtained, the highest trans-resveratrol content in the transgenic young leaves of purple-fleshed "Jashim" was 2.11 μgg-1 fresh weight and that in the microtubers in vitro of purple fleshed "Jashim" was 8.31 μgg-1 fresh weight. This amount of resveratrol may have a positive biological effect on human health.