Livestock Wastewater shall cause a high concentration of organic matter and nutrients such as rivers because of the lake and groundwater contamination, such as the accumulation of nutrients in the soil contained in the manure, livestock wastewater containing large amounts of organic matter that will flow to the river or appeal If eutrophication, and comfortable living environment to cause harm, such as odor and pest damage and can. Organic waste and organic waste, such as the world has a direct interest in acquiring the available resources and the development of clean energy from waste is a growing desire, is expected to contribute to the environment from waste materials industry growth by developing innovative technologies such as direct electrical energy production. In the case of livestock waste water and high concentration of organic material in addition to containing ammonia nitrogen, nitrate nitrogen for nitrification is created due to the electron acceptor is used as the fuel cell according to this has been reported to decrease the efficiency of electricity production. Therefore, to derive the electricity production efficiency due to organic concentration and ammonia nitrogen concentration in order to apply a microbial fuel cell (MFC) livestock wastewater treatment process in this study, and to derive the energy production potential with livestock waste water through the study. Lab. scale Reactor fabrication and operation to try to derive the reaction factor of the optimum operating conditions in accordance with the livestock wastewater applied through the evaluation of trends and removal efficiency of organic matter and nutrients in the microbial fuel cell. In addition, from the final research results, I try to present the direction of future research for the improvement of application possibilities and microbial fuel cell power generation efficiency of microbial fuel cell in the livestock wastewater treatment facilities.

A prototype of the GMT FSM has been developed to acquire and to enhance the key technology – mirror fabrication and tiptilt actuation. The ellipsoidal off-axis mirror has been designed, analyzed, and fabricated from light-weighting to grinding, polishing, and figuring of the mirror surface. The mirror was tested by using an interferometer together with CGHs, which revealed the surface error of 13.7 nm rms in the diameter of 1030 mm. The SCOTS test was employed to independently validate the test results. It measured the surface error to be 17.4 nm rms in the diameter of 1010 mm. Both tests show the optical surface of the FSMP mirror within the required value of 20 nm rms surface error.

새로운 베지클인 glyceryl citrate/ lactate/ linoleate/ oleate를 이용한 수중유형 형태의 아스타잔틴 나노에멀젼에 대해 항산화 효과, 세포 생존력, 단백질과 관련한 효소의 영향, 피부 침투도 그리고 피부에 대한 보습 및 탄력 등의 약용화장품적인 측면에서의 전반적 연구를 실시하였다. 항산화력 및 세포 생존력에 대해선 각각 DPPH법과 MMT assay를 이용하여 측정하였다. 아스타잔틴 나노에멀젼에 대한 또 다른 성질은 2D-Page를 이용한 단백질 분석 및 컨포칼, in-vivo 테스트를 통해 측정하였다. 본 연구를 통해, 아스타잔틴을 포함하는 나노에멀젼은 MMP발현에 관련한 단백질 억제 및 세포외 기질의 분해를 막고 라디칼의 소거에 매우 우수한 결과를 보였다. 종전의 레시친을 이용한 나노에멀젼 보다는 새로운 베지클을 이용한 아스타잔틴 나노에멀젼의 피부 침투가 매우 효과적임을 CLSM을 통해 측정하였다. 또한 28일 동안의 한국 성인 여성 11명을 통한 보습 및 탄력 인비보 테스트에서 우수한 효과를 확인할 수 있었다.

The transcription factor, early growth response protein 1 (EGR1), act as immediate early response genes to control various cellular and reproductive events. Egr1-deficient female mice show infertility by anovulation resulting from luteinizing hormone-β (LH-β) subunit deficiency. While ovulation, fertilization and embryo development normally occur in Egr1-deficient mice treated with a superovulation regime to rescue LH deficiency, embryo implantation was completely failed. The morphology and ultrastructure of uterine tissues were observed by light and transmission electron microscopy during the peri-implantation period in Egr1-deficient mice. To examine alterations in cellular organelles, the uterine horns were fixed with 2.5% glutaraldehyde and postfixed with 1% osmium tetroxide in PBS. After dehydration and infiltration, the samples were embedded in Epon 812. Semi-thin sections 0.5 μm thick were cut with an ultramicrotome and stained with toluidine blue for light microscopy. Thin sections were cut with a diamond knife of the ultramicrotome and placed on copper grids. The sections were double stained and examined under a transmission electron microscope. The height of luminal epithelial cells was decreased and the polarity was poorly differentiated in the Egr1-deficient comparing to the wild mice. The abundant mucinous materials were observed in the surface of luminal epithelial cells of the Egr1-deficient. It was confirmed the microarray and real time qPCR data. The luminal epithelial cells of wild mice had many dense lipophilic granules and healthy mitochondria, but not in the Egr1-deficient. It may related to production and secretion of steroid hormones and prostaglandins in the luminal epithelial cells for successful implantation. These results show that Egr1 is a critical transcription factor to fine-tune subcellular morphological and functional changes for the receptive phase of peri-implantation period of uterine tissue in mice.

AtRabG3b and CaMsrB2 genes incorporated into pPZP vetor were transformed to Korean soybean cultivar Kwangan using highly efficient transformation system. AtRabG3b gene plays a positive role in xylem development in Arabidopsis and 64 transgenic plants were produced. CaMsrB2 gene is known to confer drought tolerance in rice and 63 transgenic plants were produced. As a result of PPT leaf painting assay, about 20% of transformation efficiency was observed from 2 times of inoculation. These transgenic plants were confirmed for gene introduction using PCR. Currently, the copy number and the gene expression is investigating using qRT-PCR and RT-PCR. Moreover, 62 lines and 53 lines of T1 seeds from AtRabG3b and CaMsrB2, respectively, were sown in GMO field.

The PepMoV has been considered the most frequently detected potyvirus. When it co-infects with CMV or PMMoV, it gives severe impact to total pepper harvest in Korea. Since F1 hybrid that resistant to PepMoV has not been developed, we have developed transgenic peppers using Agrobacterium-mediated transformation with a Hc-Pro gene of the PepMoV. A large number of GM peppers were tested for resistance to the PepMoV, and after consequent self-crossing up to T4 generation, a highly tolerant pepper to PepMoV called T20 was selected. So far, BC4F1 lines have been selected by back-crossing with 4 elite lines through a breeding program. Very recently, based on molecular analysis, we have selected another event, #10-2, which is also resistant to PepMoV. Horticultural difference was investigated for both GM lines, #T20 and 10-2, and no significance was found comparing to non-GM lines.

Soybean mosaic virus (SMV), a member of Potyviridae family, is one of the most typical viral diseases and results in yield and quality loss of cultivated soybean. Due to the depletion of genetic resources for resistance breeding, a trial of genetic transformation to improve disease resistance has been performed by introducing SMV-CP and HC-Pro gene by RNA interference (RNAi) method via Agrobacterium-mediated transformation. Transgenic plants were infected with SMV strain G5 and investigated the viral response. As a result, two lines (3 and 4) of SMV-CP(RNAi) transgenic plants and three lines (2, 5 and 6) of HC-Pro(RNAi) transgenic plants showed viral resistance. In genomic Southern blot analysis, most of lines contained at least one T-DNA insertion in both SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. Subsequent investigation confirmed that no viral CP and HC-Pro gene expression was detected in two SMV-resistant lines of SMV-CP(RNAi) and three lines of HC-Pro(RNAi) transgenic plants, respectively. On the other hand, non-transgenic plants and other lines showed viral RNA expression. Viral symptoms affected seed morphology, and clean seeds were harvested from SMV-resistant line of SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. In addition, strong viral gene expression was detected from seeds of SMV-susceptible non-transgenic plants and SMV-susceptible transgenic lines. When compared the viral resistance between SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants, soybean transgenic plants with the HC-Pro gene using RNAi strategy showed much stronger and higher frequency of viral resistance.

Mungbean (Vigna radiata) is a fast-growing, warm-season legume crop that is primarily cultivated in developing countries of Asia. We constructed a draft genome sequence of mungbean to facilitate genome research into the subgenus Ceratotropis and to enable a better understanding of the evolution of leguminous species. The draft genome sequence covers 80% of the estimated genome, of which 50.1% consists of repetitive sequences. In total, 22,427 high confidence protein-coding genes were predicted. Based on the de novo assembly of additional wild mungbean species, the divergence of what was eventually domesticated and the sampled wild mungbean species appears to have predated domestication. Moreover, the de novo assembly of a tetraploid Vigna species (Vigna reflexo-pilosa var. glabra) provided genomic evidence of a recent allopolyploid event. To further study speciation, we compared de novo RNA-seq assemblies of 22 accessions of 18 Vigna species and protein sets of Glycine max and Cajanus cajan. The species tree was constructed by a Bayesian Markov chain Monte Carlo method using highly confident orthologs shared by all 24 accessions. The present assembly of V. radiata var. radiata will facilitate genome research and accelerate molecular breeding of the subgenus Ceratotropis.

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.

Soybean mosaic virus (SMV), a member of Potyviridae family, is one of the most typical viral diseases and results in yield and quality loss of cultivated soybean. Due to the depletion of genetic resources for resistance breeding, a trial of genetic transformation to improve disease resistance has been performed by introducing SMV-CP and HC-Pro gene by RNA interference (RNAi) method via Agrobacterium-mediated transformation. Transgenic plants were infected with SMV strain G5 and investigated the viral response. As a result, two lines (3 and 4) of SMV-CP(RNAi) transgenic plants and three lines (2, 5 and 6) of HC-Pro(RNAi) transgenic plants showed viral resistance. In genomic Southern blot analysis, most of lines contained at least one T-DNA insertion in both SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. Subsequent investigation confirmed that no viral CP and HC-Pro gene expression was detected in two SMV-resistant lines of SMV-CP(RNAi) and three lines of HC-Pro(RNAi) transgenic plants, respectively. On the other hand, non-transgenic plants and other lines showed viral RNA expression. Viral symptoms affected seed morphology, and clean seeds were harvested from SMV-resistant line of SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. In addition, strong viral gene expression was detected from seeds of SMV-susceptible non-transgenic plants and SMV-susceptible transgenic lines. When compared the viral resistance between SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants, soybean transgenic plants with the HC-Pro gene using RNAi strategy showed much stronger and higher frequency of viral resistance.

Recently, detection of esophageal diverticula by neck ultrasonography has been increasing. In particular, esophageal diverticula near the thyroid gland can be diagnosed as thyroid nodule on ultrasonography. To date, 13 cases of ultrasonographic features of Killian-Jamieson (K-J) diverticula have been reported. We report on four cases of K-J diverticula and discuss clinical characteristics, including ultrasonographic findings, in comparison with previously reported cases. Awareness of ultrasonographic finding of esophageal diverticula, a hypoechoic nodule containing echogenic foci found in the posterolateral aspect of the thyroid, is most important for making a differential diagnosis of K-J diverticulum from a thyroid nodule.

Totally, 26 collections, 17 from Korea and 9 from China, were investigated for their sequences of 5S rDNA, especially the non-transcribed spacers (NTSs). Sequences of 5S rDNA were isolated by PCR using the primers, 5s-rRNA1 and 5s-rRNA2. Genomic DNA PCR produced single amplification of 300, 330, or 350 base pair fragments. Sequence analysis revealed that all inserts contained the part of 5S rDNA gene sequence and the full length of the NTS region. Three different sizes of the fragments were confirmed due to different size of NTS and their length were 300bp, 330bp and 350bp, respectively. Among 17 Korean foxtail millets tested, 14 collections showed single 300bp amplification. Longest fragment amplification, 350bp, was obtained only from the foxtail millet from China origin, even though 2 of them include 300bp fragment. CLUSTALW multiple alignments of 26 foxtail millets clearly revealed 4 areas with certain degree of sequence heterogeneity (region I, II, III, IV). Among 4 boxed areas, foxtail millet genotypes from China have distinct insertion especially in region III. Five of them have extra insertion of sequence and their additional sequences were either 45 or 48 base pair. Three Korean foxtail millets have 32 bp insertion. Other 8 Korean collections have short insert sequences (6 to 8 bp), 3 with 8 bp and 5 with 6 bp. In addition to insert, deletion sequences were also confirmed as major deletion was observed in region II of Chinese collection. The size of deletion was 7 bp long. According to phylogenic tree constructed using MEGA4 program, clear grouping was not revealed. To obtain more convincing results various collections from many countries should be obtained and analyzed to distinguish different germplasm from different origin.

Citrus is one of the major fruits produced in Korea. There are about 20 species mainly grown in Jeju Island, Korea. Four representative species, which are quite different in the shape of leaf and the taste of fruit, were selected and were used to profile the transcriptomes. These species are ‘Miyagawa Wase’ (C. unshiu Marcov.) satsuma mandarin, ‘Kiyomi’ (C. unshiu Marcov. × C. sinensis) mandarin hybrid, ‘Dangyuja’ (C. grandis) and ‘Natsudaidai’ (C. natsudaidai). Classification of the up-regulated and down-regulated genes using the Cluster of Orthologous Groups of proteins (COG) database reveals that the number of genes included in each group differed significantly among the four species. Several genes that showed significant differences in expression on the microarray were selected and their expression patterns were examined by reverse transcription- ploymerase chain reaction. Metabolic genes such as tyrosine decarboxylase and β-glucosidase ligase were found to be highly expressed in Miyagawa Wase, relative to other species. On the other hand, the expression level of mannose phosphate isomerase was lower in Miyagawa Wase. An efflux pump gene was found to be up-regulated in Kiyomi, whereas cinnamyl-alcohol dehydrogenase was down-regulated. β-carotene 15,15’-dioxygenase, which is involved in the vitamin metabolism, was up-regulated in Natsudaidai. Interspecific differentiations of gene expression are analyzed in terms of the metabolic pathways and their possible roles in citrus species.

Protein disulfide isomerase (PDI) is a chaperone protein that involves in oxidative protein folding by acting as catalysts and folding assistants in the endoplasmic reticulum (ER). Genome database showed that rice contains three PDI-like genes. But, their functions and subcellullar localization are not clearly identified. Here, we show possible functions of rice PDI (OsPDI) during seed development. Seeds of OsPDI T-DNA insertion mutants which were identified by genomic DNA PCR and western blot display chalky phenotype. Electron microscope analysis revealed that endosperms of the OsPDIL1-1Δ mutant show imperfect packing of round starch granules, causing floury-white color. Abnormal form of protein body I (PB-I) containing prolamin and thick aleurone layer were also observed in the OsPDIL1-1Δ mutants. Protein content per seed was significantly low in the OsPDIL1-1Δ mutant. However, free sugar content was high in the OsPDIL1-1Δ mutant seed. Northern and western blot analyses showed that during seed development, OsPDI protein is steadily accumulated in the seed until maturation while its transcript level was highest at 10 days after flowering and rapidly decreased to basal level. In addition, OsPDI strongly interacts with cysteine protease OsCP1 and chaperone BiP protein accumulates in OsPDIL1-1Δ mutant. Besides, proteomic analysis of the OsPDIL1-1Δ mutant seed showed that OsPDI is post-translationally regulated and its loss causes accumulation of many types of seed proteins. Our results indicate that OsPDI plays a critical role in seed development through its regulatory activity for various proteins.

Grain yield, one of the most important agronomic traits, is greatly affected by architecture in rice. Here, we show that an OsPrMC3, a rice PrMC3 orthologue with a lipase or esterase domain, involves in yielding by tillering. Phenotypic analysis of T-DNA insertion mutant revealed that it has high number of tillers than wild type although height and leaf width are shorter and narrower than wild type. Size and branch number of panicle were greatly reduced in the mutant, which resulted in significant decrease of seed number per panicle and dry weight of the seeds. OsPrMC3 is highly expressed in the leaf during the early stage of development. However, it is mainly expressed in mature seed and root after flowering although its expression is detected in all of the tissues. Our result indicates that OsPrMC3 involves in leaf growth and tillering during vegetative growth and also seed development after flowering, suggesting its crucial regulatory role in yielding

CGMMV (cucumber green mottle mosaic virus) infects many cucurbitaceae species causing mosaic symptoms, yellowish leaf, and fruit deterioration. Several isolates of CGMMV from Korea, Israel, Japan, Greece and Spain have been characterized. In fact, CGMMV outbreaks have caused huge losses in the total yields of cucurbitaceous crops in Korea. Unfortunately, no genetic source is available for the resistance against CGMMV infection, and therefore, the breeding management offers no access to a solution yet. Thus, an alternative was to transform the viral gene such as CGMMV-CP into crops to induce tolerance against CGMMV. Here, we present the successful transformation of watermelon rootstock (gongdae) by Agrobacterium-mediated transformation with the CGMMV-CP gene. We obtained 29 independent T0 watermelon rootstock plants from the transformation of 7,887 explants. T0 plants were self-crossed and T1 plants were screened by the virus infection. Those gongdaes tolerant to virus were selected continuously, and BC2T1 and T3 generation were obtained. We have also obtained watermelons of BCF generation by crossing GM gongdae to watermelon line and those BCF watermelones were strongly tolerant to CGMMV.

To produce abiotic stress resistant transgenic cucumber, the cotyledonary node explants of cucumber (c.v. Eunsung) were inoculated with A. tumefaciens strain EHA105 containing the binary vector (pPZP211) carrying Nit gene. The 491 explants inoculated with bacterium solution for 30 min were maintained on 50 mg/L paromomycin contained shoot induction (SI) medium for first 2 weeks and then subcultured on 100 mg/L paromomycin to obtain transgenic adventitious shoots for 4 x 14 days. So far, 5 plant were selected, and then acclimated in soil. Of them, 3 transgenic plants with Nit gene were confirmed by Southern blot analysis.

R genes are a key component of genetic interactions between plants and biotrophic bacteria and are known to regulate resistance against bacterial invasion. The most common R proteins contain a nucleotide-binding site and a leucine-rich repeat (NBS-LRR) domain. Some soybean NBS-LRR genes have also been reported to function in disease resistance. A total of 319 genes were determined to be putative NBS-LRR genes in the soybean genome. The number of NBS-LRR genes on each chromosome was highly correlated with the number of disease resistance QTL in the 2-Mb flanking regions of NBS-LRR genes. In addition, the recently duplicated regions contained duplicated NBS-LRR genes and duplicated disease resistance QTL, and possessed either an uneven or even number of NBS-LRR genes on each side. The significant difference in NBS-LRR gene expression between a resistant near-isogenic line (NIL) and a susceptible NIL after inoculation of Xanthomonas axonopodis pv. glycines supports the conjecture that NBS-LRR genes have disease resistance functions in the soybean genome. The number of NBS-LRR genes and disease resistance QTL in the 2-Mb flanking regions of each chromosome was significantly correlated, and several recently duplicated regions that contain NBS-LRR genes harbored disease resistance QTL for both sides. In addition, NBS-LRR gene expression was significantly different between the BLP-resistant NIL and the BLP-susceptible NIL in response to bacterial infection. From these observations, NBS-LRR genes are suggested to contribute to disease resistance in soybean. Moreover, we propose models for how NBS-LRR genes were duplicated, and apply Ks values for each NBS-LRR gene cluster.

Gene expression profiles can serve as a valuable reference for deciphering gene functions. We exploited the potential of whole genome microarrays to measure the temporal expression profiles of rice genes in 13 stages of reproductive development. We could profile expression of 17,676 genes in at least one of the tissues. Differential expression analysis with compare to leaf and preceding stages of development revealed reproductive stage-preferential/-specific genes. we identified 35 genes expressing specifically during panicle and seed development. The metabolic/hormonal pathways and transcription factor families playing key role in reproductive development were elucidated after overlaying the expression data on the public databases and manually curated list of transcription factors, respectively. During floral meristem differentiation (P1cm) and male meiosis (P5cm), the genes involved in jasmonic acid and gebbellin biosynthesis were significantly upregulated. F11DAP stage of seed, containing enlargement organ, exhibited enrichment of transcripts involved in starch or sucrose biosynthesis. Genes regulating auxin biosynthesis were induced during early seed development. We validated the stage-specificity of regulatory regions of two panicle-specific genes, AK072471, Os08g0538700, and AK121412, an early seed-specific gene, in transgenic rice. The data generated here provides a snapshot of the underlying complexity of the gene networks regulating rice reproductive development.