Adipose tissue-derived mesenchymal stem cells (ASCs) are very interesting in several laboratory animals and humans because they are easy to harvest and expand to generate millions of cells from a small quantity of fat. ASCs are known as useful materials for clinical applications in human cell therapy and as a donor cell in somatic cell nuclear transfer (SCNT). Here, we investigated if 1) minipig ASCs can be isolated, self-renewed and differentiated into multiple tissue lineages, 2) ASCs can be a suitable donor cell type for generation of cloned pig. In order to isolate ASC, adipose tissues were collected from inguinal region of a 6-year-old female minipig. The ASCs were attached to the culture dish with a fibroblast-like morphology. They expressed cell-surface marker characteristics of stem cell, underwent osteogenic, adipogenic, myogenic, neurogenic and chondrogenic differentiation when exposed to specific differentiation-inducing conditions. To investigate its potential as donor cell for cloning, we respectively carried out SCNT using ASC, adult skin fibroblast (ASF) and fetal fibroblast (FF) derived from same minipig. The ratio of blastocysts to 2-cell embryos and total cell number of blastocysts were monitored as experimental parameters. In results, cleavage and developmental competence to blastocysts rate showed no significant difference among the three groups. On the other hand, total cell numbers of blastocysts derived from ASC and FF were significantly higher than in ASF (89±7.9 and 105±5.5 vs. 57.5±5.2, respectively). Our results demonstrated that ASC have potential compared to ASF and FF in terms of the in vitro development and blastocyst formation ability. In further study, we will investigate the in vivo developmental ability of ASC as donor cell for pig cloning. * This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), Institute for Veterinary Science, the BK21 program, TS Corporation and Optifarm Solution.

Insects or insect remains found in beer are one of major issues in consumer claim. Accurate estimation of inflow time isa critical factor for the settlement of such claims related with beer-contaminating insects but no reliable methods have been developed. In an attempt to establish a molecular marker-based diagnostic method, the degradation rates of 18S rRNA genes in the insectssoaked in 500 ml beer were investigated by quantitative real-time PCR (qPCR) over one month period at room temperature. Among the six insect species tested, the house fly (Musca domestica) and honey bee (Apis mellifera) revealed high correlations (r2=0.974-0.990) between the degradation of 18S rRNA gene and inflow time. In these insects, statistically significant distinction was possible between the samples stored in beer less than 14 days and more than 14 days. Other insects, including the fruit fly, common house mosquito, German cockroach and Indian meal moth, displayed poor correlations, which appeared attributed to the inefficient genomic DNA extraction likely due to small sample size or disintegration of body parts during storage in beer. With proper improvement in DNA extraction, this 18S rRNA-based diagnostic method would be applicable for estimating the inflow time of beer-contaminating insects.

The purpose of this study was to determine the effects of heat appli˗ cation on the immune activities of the human body. To exam, further˗ more, the immune effect from the healthy volunteer(male:15, female:15) by monitoring changes of immune substances such as various leukocytes[total white blood cell(WBC), eosinophil, neutrophil, basophil, monocyte, and lymphocyte], a comparative study with warm water immersion(40.8±0.3℃) and infrared(250W) was carried out. The plasma analysis showed that the count of white blood cell, eosinophil, and neutrophil were elevated in warm water immersion- or infrared˗ stimulated group compared with control group. However, the count of basophil was decreased in both warm water immersion- and infrared-stimulated group than control group. Therefore, these results suggest that the thermostimulation improved immune activity.

This study was conducted to examine the effect of IRES controlled reporter gene on screening and production of recombinant human erythropoietin (EPO) proteins from cultured CHO cells. The cDNA was cloned for EPO from human liver cDNA. Using site-directed mutagenesis, we generated recombinant human EPO (rhEPO) with two additional N-glycosylations (Novel erythropoiesis-stimulating protein: NESP). Wild type hEPO and NESP were cloned into expression vectors with GFP reporter gene under regulatory control of CMV promoter and IRES so that the vectors could express both rhEPO and GFP. The expression vectors were transfected to cultured CHO-K1 cells. Under microscopy, expression of GFP was visible. Using supernatant of the culture, ELISA assay, immunocytochemistry and in vitro assay using EPO dependant cell line were performed to estimate biological activity to compare the production characteristics (secretion levels, etc.) between rhEPO and NESP. The activity of NESP protein, obtained by mutagenesis, was described and compared with its rhEPO counterpart produced under same conditions. Although NESP had less secretion level in CHO cell line, the biological activity of NESP was greater than that of rhEPO. These results are consistent with previous researches. We also demonstrated that rhEPO and GFP proteins expressed simultaneously from transfected CHO cell line. Therefore we conclude that use of GFP reporter gene under IRES control could be used to screen and produce rhEPO in cultured CHO cells.

We present BV CCD photometry for the open clusters Czernik 24 and Czernik 27. These clusters have never been studied before, and we provide, for the first time, the cluster parameters; reddening, distance, metallicity and age. Czernik 24 is an old open cluster with age 1.8 ± 0.2 Gyr, metallicity [Fe/H]=-0.41 ± 0.15 dex, distance modulus (m-M)0=13.1 ± 0.3 mag (d=4.1 ± 0.5 kpc), and reddening E(B-V)=0.54 ± 0.12 mag. The parameters for Czernik 27 are estimated to be age=0.63 ± 0.07 Gyr, [Fe/H]=-0.02 ± 0.10 dex, (m-M)0=13.8 ± 0.2 mag (d=5.8 ± 0.5 kpc), and E(B-V)=0.15 ± 0.05 mag. The metallicity and distance values for Czernik 24 are consistent with the relation between the metallicity and the Galactocentric distance of other old open clusters. We find the metallicity gradient of 51 old open clusters including Czernik 24 to be Δ[Fe/H]/ΔRgc = -0.064±0.009 dex kpc-1.

Experiments were carried out to develop an optimal nutrient solution for the single-stemmed rose (Rosa hybrida L.) 'Red velvet' in a closed aeroponic system. Plants were grown in 1/3, 1/2, 1, or 3/2 strength of the nutrient solution of National Horticultural Research Station in Japan (NHRS). Significantly less changes of pH and EC (dS·m-1) in the drainage were observed in 1/2 strength treatment as compared to other treatments. The NO3-N, K, Ca, and Mg concentrations in the drainage solution of 1/2 strength treatment were maintained at optimal levels. These results indicated that the rose uptakes of both nutrients and water was more stable than those in other concentration. The concentration of macronutrients in nutrient solution were adjusted based on the ratio of nutrient:water (n/w) taken up by plants grown in the 1/2 strength solution. The composition of the new solution (classified the University of Seoul (UOS) solution) was as follow; NO3-N 8.8, NH4-N 0.67, P 2.0, K 4.8, Ca 4.0, Mg 2.0 me·L-1. To further evaluate new solution on crop growth, the rose 'Red Velvet' was grown again in l/2, 1, and 2 strength UOS solution to compare with 1.0 strength PBG (proefstion voor bloemisterij en glasgroenpe) solution. Overall the plant growth, including the stem length and number of five-leaflet leaves was higher in 1.0 strength of UOS solution than other treatments. Results presented in this study indicate that the nutrients in the UOS solution are well balanced for the single-stemmed rose in the closed aeroponic system.

장 건강에 유익한 프리바이오틱스 소재를 개발하기 위하여 배변을 촉진하는 효능을 지닌 붉은팥을 식품 발효에 이용되는 Bacillus subtilis KCCM 11965P로 발효하여 다음과 같은 결과를 얻었다. 붉은팥의 일반성분은 회분 3.35±0.04%, 조단백질 21.1±0.19%, 조지방 0.35±0.02% 함 유되었다. 붉은팥 원물 1%, 3%, 5%와 Bacillus subtilis KCCM 11965P 3% (v/v)를 접종하여 0, 24, 48, 72시간 배양 하였다. 배양액의 총균수를 측정한 결과 붉은팥 원물을 3% 첨가한 후 72시간 배양군에서 Bacillus subtilis KCCM 11965P 발효가 가장 적합하였다. 발효 시간에 증가함에 따라 총 폴리페놀 함량과 DPPH 라디칼 소거활성이 증가하였다. Protease 활성은 붉은팥 원물 5% 첨가한 후 72시간 배양한 군(2.69±0.003 unit/mL)에서 활성이 가장 높았다. 발효시간과 붉은팥 원물 첨가 농도가 증가함에 따라 α -amylase 활성도 증가하였으며, 붉은팥 원물 5% 첨가한 후 72시간 배양한 군에서 0시간 배양군(1.0±0.1 unit/mL) 보다 26.0±0.2 unit/mL로 증가하였다. Bacillus subtilis KCCM 11965P로 72시간 배양한 후 유리아미노산을 측정한 결과 leucine은 붉은팥 원물 5% 첨가한 0시간 배양군 5.22 mg/L 에서 67.59 mg/L로 증가하였으며, 비필수아미노산인 tyrosine은 5% 첨가 0시간 배양군 10.08 mg/L에서 259.35 mg/L로 증가하였다. 이와 같이 Bacillus subtilis KCCM 11965P로 붉은팥을 발효하면 항산화 활성, protease 효소활성, 및 α-amylase 효소 활성이 증가하였으며, 유리아미노 산과 유기산이 증가하였다. 붉은팥을 발효하는데 Bacillus subtilis KCCM 11965P가 적합할 것으로 판단되며, 붉은팥은 프로바이오틱스를 활성화시켜 장 건강을 증진시킬 수 있는 프리바이오틱스 소재로 개발할 수 있는 가능성을 시사 하였다.

The purpose of this study was to investigate the improvement of growth in Israeli carp (Cyprinus carpio), and the cross experiment was carried out with two strains of Israeli carp. Four combinations of Israeli carp from Jeonbuk fisheries farm and Songpu mirror carp from Heilong Jiang, China (KK; Jeonbuk ♀ × Jeonbuk ♂, KC; Jeonbuk ♀ × China ♂, CC; China ♀ × China ♂ and CK; China ♀ × Jeonbuk ♂) were developed and reared. Body length, body weight and condition factor were determined at 20, 40, 60 and 170 days post-hatch (DPH). The results showed that there were differences in growth rate of the four groups. Body length of four groups were CK > CC > KC > KK and body weight were CC > CK > KC > KK at 170 DPH. The growth perfomance of four groups were statistically significant difference (P<0.05). During the rearing, CC group had longer length and higher weight at 170 DPH compared to other three groups and also condition factor was highest in the CC group, but there was no significant difference in a survival rate. These results indicated that the growth performance mainly depended upon brooder combination but survival rate could not significantly affect brooder.

Background : Gentinae Macrophyllae Radix is one of the traditional medicines originated from the roots of multiple plants, Gentiana macrophylla Pall., Gentiana straminea Maxim., Gentiana crassicaulis Duthie ex Burkill and Gentiana dahurica Fisch., in Gentianaceae. Multi-origin traditional medicine usually has adulteration problem based on the morphological similarity and/or misunderstanding the species. Therefore, accurate and reliable identification criteria to ensure drug safety and quality is necessary. Methods and Results : We collected four original species of Gentinae Macrophyllae Radix from plantations and markets in China and Korea. DNA barcoding with four barcoding markers (Internal Transcribed Spacer (ITS), rbcL, trnL intron, trnL-F intergenic sapcer) was performed. Intra-specific variation was observed in ITS nucleotide sequence however, successfully distinct four original species based on the nucleotide discrepancy while trnL intron has no difference. trnL-F intergenic spacer has two transitions(T→C and A→G) sites only in G. crassicaulis and rbcL shows one transition(C→T) site in G. dahurica and G. macrophylla. Phylogenetic relationship analysis of the Gentinae Macrophyllae Radix revealed two major clades – clade I including three groups, G. macrophylla, G. straminea and G. dahurica, and clade II including G. crassicaulis. This aspects was shown more clear with multi-region combined analysis. Conclusion : DNA barcoding will be accurate and powerful criteria for the analysis the origin of Gentinae Macrophyllae Radix. However, single region analysis might be deficient such as trnL intron, rbcL and trnL-F intergenic spacer results in this research. Multi-region combined analysis based on the multiregional DNA barcode markers will be overcome the disadvantage and also increase the precision.

Background : The advancement of next-generation sequencing technology dramatically reduces the cost for sequencing and it contributes to create a new research environment that utilizes large amount of genome sequences to answer many biological questions. With this new research trend, reference genome sequences of several major crops have been released to the research community and utilized in various researches in agriculture. Coupled with molecular breeding technology, NGS based genome research will possibly allow selecting a new plant material possessing useful traits in early stage and efficiently developing a superior cultivar. Methods and Results : The objectives of this research are to collecting various genetic variations (SNPs, indels and TE mediated variations) in major and minor crops, to develop molecular markers using NGS based genomic data (resequencing, GBS, transcriptome), and to develop a visualization tools to enhance the utility of the NGS data. Currently major analysis pipelines have been developed to detect SNPs, indel and polymophic SSRs using whole genome and transcriptome data, and a pipeline for identification of MITE insertion polymorphism is under development. In addition to that, for orphan crop, we also implemented an efficient and robust method to assemble a complete chloroplast, mitochondria and 45S rDNA using low coverage whole genome data in order to develop an inter- and intra-specific molecular barcode markers. Conclusion : NGS provide a new level of researches in many crop plants. Large amount of genomic information provides an opportunity to understand domestication and genetic variations, and to develop a better crop for future.

Background : This study was carried out to survey the effect of IBA (indole-3-butric acid) concentration on root development characteristics in Viburnum dilatatum cuttings. Methods and Results : The cuttings were collected on July 28, 2016 from plants growing in Korea National Arboretum. Cuttings were treated with different IBA concentrations [0 (control), 10, 100, 1000, 2000 ㎎⋅L-¹] for 10 seconds. Rooting parameters (root number, the longest root length, root fresh weight, root dry weight, rooting percentage, and survival rate) were observed at 8 weeks after cuttings. Rooting percentage was 43, 33, 17, 47, and 40 % at 0, 10, 100, 1000, and 2000 ㎎⋅L-¹ of IBA, respectively. Root number (13.4) was highest under 1000 ㎎⋅L-¹ IBA. Root length (9.4 ㎝) and Root weight (fresh and dry) were highest under 1000 ㎎⋅L-¹ IBA. Conclusion : According to the results, 1000 ㎎⋅L-¹ IBA was the best treatment for increasing percentage rooting percentage and other root development characteristics of V. dilatatum cuttings.

Background : Clematis trichotoma is a deciduous climber belonging to the family Ranunculaceae. This study was carried out to survey the effect of shading levels on growth characteristics in Clematis trichotoma seedling. Methods and Results : Seeds were collected from plants growing in the Mt. Kariwang Jeongseon, Gangwon-do in October 2013, and they were sown to 96 cell plug tray filled with Peatmoss (TKS-2) soil at March, 2015. The experiment was performed with four different shading levels (0, 30, 60, 90%) at July, 2015. According to the experiment, plant height was the highest under 90% of shading. It was found that fresh weight and dry weight of clematis trichotoma were the highest under 90% of shading. The leaf number was the highest under 30% of shading. The leaf number decreased as the shading level increased. Root number and length were the highest under 90% of shading. Conclusion : According to the results, Clematis trichotoma seedling showed the highest growth under 90% shading.

Background : Scrophularia koraiensis is a herbaceous perennials belonging to the family Scrophulariaceae. This study was carried out to survey the effect of LED on root develop characteristics in Scrophularia koraiensis seedling. Methods and Results : Seeds were collected from plants growing in the Mt. Kariwang Jeongseon, Gangwon-do in September 2014, and they were sown to 128 cell plug tray filled with Peatmoss (TKS-2) soil at March, 2015. The experiment was performed with three different LED (Blue, Red, Blue + Red) at July, 2015. Morphological characteristics of root (total root length, root projet area, root surface area, root diameter and root volume) were analyzed with WinRHIZO software. Seedling root growth of scrophularia koraiensis was surveyed to be the highest at the Blue + Red LED in all measuring. Total root length was measured high in the order of Blue + Red, Red, Blue LED. Conclusion : According to the results, Scrophularia koraiensis seedling showed the highest root growth in Blue + Red LED.

The isolated single coronary artery is a rare congenital anomaly, in which both coronary arteries arise from a solitary ostium. Diagnosis of coronary anomalies and identification of the exact anatomy of coronary arteries has significant clinical importance, hence, myocardial ischemia or sudden cardiac death is usually related to its course of anomalous coronary artery. Most patients with a single coronary artery are asymptomatic and have normal electrocardiogram and negative stress tests. However, if the patient has other structural abnormalities, for example, ventricular hypertrophy, the exam is determined. This report describes a case of single coronary artery, where the right coronary artery originated from the distal left circumflex artery in a patient with hypertrophic ardiomyopathy.

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.

Olive flounder (Paralichthys olivaceus) is one of the commercial important flatfish species in Korea. The ocular signal transduction pathway is important in newly hatched flounders because it is closely involved in the initial feeding phase thus essential for survival during the juvenile period. However, the study of gene expression during ocular development is incomplete in olive flounder. Therefore we examined the expression analysis of specifically induced genes during the development of the visual system in newly hatched flounders. We searched ocular development-involved gene in the database of expressed sequence tags (ESTs) from olive flounder eye and this gene similar to arrestin with a partial sequence homology. Microscopic observation of retinal formation corresponded with the time of expression of the arrestin gene in the developmental stage. These results suggest that arrestin plays a vital role in the visual signal transduction pathway of the retina during ocular development. The expression of arrestin was strong in the ocular system during the entirety of the development stages. Our findings regarding arrestin have important implications with respect to its biological role and evolution of G-protein coupled receptor (GPCR) signaling in olive flounder. Further studies are required on the GPCR-mediated signaling pathway and to decipher the functional role of arrestin.

Low temperature is a major factor restrict to growth and limiting productivity of rice crops. We used a cDNA microarray approach to monitor the expression profile of rice (Oryza sativa) under chilling stress and identified 20 chilling inducible genes in previously study. Ten such genes encoding bHLH, metal transporter and, zinc finger protein with unknown functions showed a significant change in expression under various abiotic stresses. Among them, OsCHI1 (Os07g15460), OsCHI2 (Os02g43660), and OsCHI3 (Os01g61160), were selected for further study. They have structural features such as metal-binding signature sequences in their protein sequences, and OsCHI genes were expressed in root of rice seedling and induced in chilling and salt or drought. Expression of OsCHI1, OsCHI3 and OsCHI2 were targeted to membrane and ER when transiently expressed in tobacco cell, respectively. The Arabidopsis (Arabidopsis thaliana) transgenic plants overexpressing showed increased tolerance to salt and drought stress in the seed germination and root elongation than that of wild type. This comprehensive study provides insight into the biological function of OsCHIs, which may be useful in understanding how rice plants adapt to unfavorable environmental conditions.

Green roof policies has helped to advance urban greening by creating green spaces on previously unused rooftops. However, there are no requirements covering matters such as greening methods, operation and maintenance obligations and many problems for sustainability and functioning of green roof and green space have been pointed out. This research is intended to clarify the contents and criteria of green roof policies currently implemented in Korea, Japan and other countries, compare them with the evaluation criteria for sustainable and public benefit rating systems and propose comprehensive indices for evaluating the multidimensional functions of green roof from the perspective of urban open space. As a result, it is found that obligation, grant, reimbursement, density bonus and tax credit are frequently used as green roof policies and 41 criteria including percentage of greening area, percentage of trees or shrubs, reduction of heat island effect, open space, landscape, and promotion of biodiversity are identified as criteria applicable to evaluating the multidimensional functions of green roof from the perspective of urban open space, respectively.