Background : As a part of ongoing research to elucidate and characterize anti-inflammatory nutraceuticals, six kinds of plant extracts (aerial part of Nepeta cataria, leaves of Lonicera maackii, leaves of Platycarya strobilacea, flower of Fagopyrum dibotrys, flowers and fruits of Solanum nigrum, stem of Physostegia virginiana) were tested for their ability to suppress inflammation. The anti-inflammatory has been studied in lipopolysaccharide (LPS)-stimulated RAW264.7 cells which cells synthesized nitric oxide (NO) from L-arginine by nitric oxide synthase (NOS). In this study, NO synthesis inhibitory activity of six kinds of plant extracts on LPS-stimulated RAW 264.7 mouse macrophages was evaluated. Methods and Results : Six kinds of plant extracts were parceled out from RDA (Rural Development Administration). RAW 264.7 cells (1.5×105 cells/well) were seeded onto 96-well plates with DMEM media containing 10% FBS and 1% antibiotics. The cells were pretreated with the extracts and LPS-stimulated cells for 24 h. Cellular NO production was stimulated by adding 1 μg/mL of LPS. After incubation, Griess reagent was used to determine NO production. Absorbance was measured at 520 nm by microplate reader. NO synthesis inhibitory activity potential of these extracts was evaluated by assessing NO production by LPS-stimulated RAW 264.7 cells in the presence. As a result, inhibition rate of NO production was about 40% of L. maackii, 33% of F. dibotrys, 23% of P. strobilacea and 17% of P. virginiana. Meanwhile, there was no significant results in aerial part of N. cataria and flowers and fruits of S. nigrum. Conclusion : From the above results, we be able to confirm that leaves of L. maackii and flower of F. dibotrys appeared dose-dependent NO synthesis inhibitory activity and leaves of P. strobilacea appeared NO synthesis inhibitory activity in low-concentration. As screening NO synthesis inhibition of six extracts, they may be a good candidate for delaying the progression of human inflammatory diseases and warrants further studies.

Background : Ginseng berry(GB) is useful not only just in growing source but also in functional food source. The ingredients of crops varies with the maturity. So, GB ingredients need to be analyse for optimal harvesting stage of GB against appropriate use. Methods and Results : This study was carried out to determine optimal harvesting stage of GB. GB was harvested 5 day periods from July 12, started harvesting when pollination was 50 days old, until August 1. GB was analysed color, ginsenosides and fatty acids using colorimeter, LC and GC, respectively. As the majority of GB increase, color of freeeze drying GB powder were changed that lightness and yellowness was increased, redness was decreased. Ginsenoside Re, Rb1 and Rb2, major ginsenoside in GB, were increased and Ginsenoside F1, Rk1 and Rg5, minor ginsenoside, were increased for a time and then decreased. Oleic acid, the main fatty acid in GB, was decreased, and linoleic acid and total fatty acid content was increased to July 27 and then decreased. Conclusion : Total ginsenosides content was the highest on August 1 and total fatty acid content was the highest July 27. As the majority of GB increase, ratio of oleic acid on total fatty acid was decreased and linoleic acid was increased. Thus, GB is that the longer a harvest period and the more useful for food source.

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.

DGCR8 is a RNA-binding protein working with DROSHA to produce pre-microRNA in the nucleus, while DICER does not only mature microRNA but also endogenous siRNAs in the cytoplasm. Here, we have produced Dgcr8 conditional knock-out mice using progesterone receptor (PR)-Cre (Dgcr8flox/flox; PRcre/+ mice, Dgcr8d/d) and demonstrated that canonical microRNAs dependent of DROSHA-DGCR8 complex are required for uterine development as well as female fertility in mice. Adult Dgcr8d/d females did not undergo regular reproductive cycle and produce any pups when housed with fertile males, whereas administration of exogenous gonadotropins induced normal ovulation with corpus luteal formation in these mice. Ovulated oocytes from Dgcr8d/d mice had comparable fertilization potentials and were normally developed to the blastocyst after fertilization as compared to those in control Dgcr8f/f mice. Interestingly, PR-Cre-dependent Dgcr8 deletion showed aberrant infiltration of acute inflammatory immune cells to female reproductive organs only when Dgcr8d/d mice were mated with male mice. With respect to uterine development, gross morphology, histology, and weight of Dgcr8d/d uterus were similar to those of control at 3-week-old age. However, multiple uterine abnormalities were noticeable at 4-week-old age when PR expression is significantly increased, and these deformities became severe onwards. Gland formation and myometrial layers were significantly reduced, and stromal cell compartment did not expand and became atrophic during uterine development in these mice. These results were consistent with aberrantly reduced cell proliferation in stromal cell compartments of Dgcr8d/d mice. Collectively, our results suggest that DGCR8 dependent-canonical microRNAs are essential for development and physiology of the uterus with respect to morphogenesis, proper immune modulation, reproductive cycle, and steroid hormone responsiveness in mice.

This study aimed to evaluate the role of D-dimer in terminal cancer patients. Among 343 incurable cancer patients, only 4 patients (2.8%) had normal D-dimer levels, and the majority (n=115, 81.0%) had levels >2.0 μg/mL. The survival time was significantly differed according to D-dimer levels with a median of 48 days when D-dimer levels were <2 μg/mL and 19 days when D-dimer levels were higher. D-dimer levels were negatively correlated with survival according to Spearman’s rank correlation analysis (coefficient -0.335, p<0.001). In terminal cancer patients, D-dimer concentrations did exhibit prog-nostic significance.

TCR subunits are members of membrane-bound receptors which allow the fast and efficient elimination of the specific fish pathogens have regulated function in adaptive immunity. Sequence structure of TCR subunits have been reported for various teleosts, but the information of each TCR subunit functional characterization through expression analysis in fish was unknown. In this study, we examined the gene expression of TCR subunits in the early developmental stages and observed transcript levels in various tissues from healthy adult olive flounder by RT-PCR. The mRNA expression of alpha subunit was already detected in the previous hatching step. But the transcripts of another TCR subunit were not observed during embryo development and increased after hatching and maintained until metamorphosis at the same level. It was found that all TCR subunits mRNAs are commonly expressed in the immune-related organ such as spleen, kidney and gill, also weak expressed in fin and eye. TCR alpha and beta subunit were expressed in brain, whereas gamma and delta were not expressed same tissue. The sequence alignment analysis shows that there are more than 80% sequence homology between TCR subunits. Because it has a high similarity of amino acid sequence to expect similar in function, but expression analysis show that will have may functional diversity due to different time and place of expression.

The innate immune system is the only defense weapon that invertebrates have, and it is the fundamental defense mechanism for fish. The innate immune response is important in newly hatched flounders because it is closely involved in the initial feeding phase, which is why it is essential for survival during the juvenile period. The expression analysis of genes involved in the innate immune response in the olive flounder (Paralichthys olivaceus) in the days after hatching is incomplete. Therefore, we have begun to examine the expression patterns of genes specifically induced during the development of the innate immune system in newly hatched flounders. Microscopic observation showed that pronephron formation corresponded with the expression of perforin-encoding gene. These results suggest that perforin plays a vital role in the innate immunity of the kidney during developmental stages. Perforin expression was strong at the start of the development of the innate immune response, and continued throughout all the development stages. Our findings have important implications with respect to perforin’s biological role and the evolution of the first defense mechanisms in olive flounder. Further studies are required to elucidate the perforin-mediated innate immunity response and to decipher the functional role of perforin in developmental stages.

Fish larvae are immediately exposed to microbes from hatching to maturation of their lymphoid organs, therefore effective innate mechanisms is very important for survival in such an environment. The key component of innate immune system, C3 is central protein of all activation pathways of the complement system, leading to inflammatory reactions, such as opsonisation, chemotaxis, and cell lysis of pathogens. Although, innate mechanisms is essential for survival in the early stage of development, little is known about defence mechanisms. In this study, the alignment analysis showed that amino acid sequence of C3 from olive flounder liver EST homologous to other known C3 sequences with 73-99% identity. Also, we examined the tissue distribution of olive flounder C3 and analyzed expression pattern from the fertilized egg until 28 days post hatching. As a result, olive flounder C3 mRNA was expressed only in the liver and the mRNA level more increased as developmental proceed during the early stage. These results may suggest that olive flounder C3 plays an important function in the early immune response of olive flounder larvae.

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.

Cathepsins are members of the multigene family of lysosomal cysteine proteinases and have regulated function in several life processes. The potential role of cathepsin F cysteine gene was expected as protease in the yolk processing mechanism during early developmental stage, but expression analysis was unknown after fertilization. The alignment analysis showed that amino acid sequence of cathepsin F from olive flounder liver expressed sequence tag (EST) homologous to cathepsin F of other known cathepsin F sequences with 87－98% identity. In this study, we examined the gene expression analysis of cathepsin F in various tissues at variety age flounder. Tissue distribution of the cathepsin F mRNA has been shown to be ubiquitous and constitutive pattern regardless of age in each group, although derived from cDNA library using liver sample. The mRNA level of cathepsin F more increased as developmental proceed during embryogenesis and early developmental stage, especially increased in the blastula, hatching stage and 3 days post hatching (dph). As a result, it may suggest that the proteolysis of yolk proteins (YPs) has been implicated as a mechanism for nutrient supply during early larval stages in olive flounder.

Olive flounder (Paralichthys olivaceus) is a most important aquaculture species in Korea. Like most marine fishes, olive flounders are stomachless at first feeding and aquired gastric function during the metamorphosis, so food was mainly digested by pancreatic enzyme from first feeding to premetamorphosis. However, comprehensive analysis of pancreatic and gastric digestive enzyme of olive flounder at early developmental period is still unclear. In the expression study of pancreatic and gastric digestive enzyme by real-time PCR at early developmental stage, pancreatic enzyme such as chymotrypsinogen 2, preproelastase 2 and 4, pancreatic protein somatomedin-B domain (PPSB) mRNA expression were initiated at first feeding and strongly expressed in the pancreas developmental stage, while gastric digestive enzyme signal was not at all detected during same period. Although, trypsinogens were secreted from pancreas and have similar amino acid sequence, trypsinogen 3 expression induction was detected both pancreas and stomach developmental stage, while trypsinogen 2 expression was significantly increased only post-metamorphosis period. Pepsinogen mRNA expression was only detected at metamorphosis according to stomach differentiation. Lipid digestive enzyme, lipase and intestine fatty acid binding protein 1 (I-FABP 1), were already reached a certain level at beginning of hatching and more increased during early developmental stage and then gradually decreased before metamorphosis. These results suggested that feed ingestion of olive flounder was exclusive charged by pancreatic digestive enyme, lipid digestive enzyme and trypsinogen 3 from first feeding and then fully swiched by gastric digestive enzyme and trypsinogen 2 from metamorphosis period.

For the study of population genetic structure with mtDNA, it is essential to measure genetic diversity at each mtDNA regions. Also, to evaluate the variation according to the each region should follow as well as to see if there are differences. In this study, we delved into the variations and dendrogram among samples of seven mtDNA regions (NDⅡ, NDⅤ, NDⅣ, NDⅣL, NDⅥ, NDⅠ, 12SrRNA) from wild Pacific abalone, Haliotis discus hannai collected in Yeosu, Korea. The region with the highest genetic variation was NDⅣ region (Haplotype diversity = 1.0000, Nucleotide diversity = 0.010823) with two to five times higher variation than the others. Furthermore, the study to see if there is a difference between the regions of samples showed that similar aspects of dendrogram in NDⅡ and NDⅠ(divergence of 90% and 87%), which forms a group with hd4, 7, 8 and 10 at bootstrap support, based on 1000 replications. Also, pair-wise FST between clusters within the regions showed high values; 0.4061 (P=0.0000), 0.4805 (P=0.0000) respectively. Therefore we can infer that it is the most efficient and accurate way to analyze the region of NDⅣ with the highest variation in addition to the regions of NDⅡ and NDⅠ, which formed clusters with high bootstrap value, for study of population genetic structure in this species.

The Egr family of zinc finger transcription factors is rapidly induced by various mitogens and regulates cell growth, differentiation, and apoptosis. While it is clear that loss of Egr1 leads to anovulatory infertility due to LHβ deficiency in female mice, molecular function of Egr1 in male reproduction has not been clearly investigated. Here, we demonstrate that Egr1 acts as an intrinsic transcription factor in Leydig cells to regulate their proliferation and steroidogenesis in the testis as well as an extrinsic factor for male reproduction via LHβ transcription in the pituitary. Egr1 is predominantly expressed in spermatogonia and Leydig cells in immature testes and later detected in some of these cell types in mature testes. The fertility potential of Egr1(-/-) male mice is relatively deteriorated even at 2 month-old age and aggravated with aging. The incidence of abnormalities of seminiferous tubules such as Sertoli cell only was dramatically increased with aging. The number and mean size of Leydig cells were significantly reduced in Egr1(-/-) testes. The impairment of Leydig cells is consistent with significant reduction in levels of testosterone and expression of factors critical for steroidogenesis such as StAR in Egr1(-/-) testes. Exogenous administration of hCG rapidly and transiently induced Egr1 expression in Leydig cells culture in vitro. hCG could reinstate reduced mean size of Leydig cells but not reduced number of Leydig cells and aberrantly low StAR expression, suggesting that Egr1 has critical functions for Leydig cell proliferation and their steroidgenesis. In addition, daily sperm production and in vitro fertilization (IVF) competence were significantly reduced, and apoptosis was facilitated in these mice. Furthermore, hCG administration to compensate for relatively low LH levels in Egr1(-/-) males could not restore the compromised reproductive phenotypes such as IVF competence and apoptosis in these mice. Interestingly, expression of Egr2, a member of Egr family, is significantly elevated in Egr1(-/-) Leydig cells suggesting that genetic compensation of Egr2 may alleviate phenotypic aberration of Egr1(-/-) male testes. Collectively, these results suggest that Egr1 act as an intrinsic transcription factor required for proliferation and steroidogenesis of Leydig cells to govern spermatogenesis in the testis.

DGCR8 is a RNA-binding protein working with DROSHA involved in critical processes for microRNA production in the nucleus. To understand function of miRNAs in the uterus, we have produced uterus-specific Dgcr8 conditional knock-out mice using two well-known Cre mouse models, anti-Mullerian hormone receptor 2 (Amhr2)-Cre and progesterone receptor (PR)-Cre. Dgcr8flox/flox;PRcre/+ mice were mainly analyzed and considered as uDgcr8 KO in this study unless otherwise indicated as Dgcr8flox/flox;Amhr2cre/+ mice. Morphological and histological analyses, embryo cultures, genomic DNA PCR, realtime RT-PCR and Western blotting were performed. uDgcr8 KO females bred with fertile males did not produce any offspring, suggesting that these mice are infertile. Vaginal smear analyses showed that these mice do not undergo estrous cycle, whereas Dgcr8flox/flox;Amhr2cre/+ mice exhibited regular estrous cyclicity. In vitro culture of 2-cell stage embryos and histological analyses for CL in uDgcr8 KO demonstrated that they can respond to gonadotrophins to ovulate healthy oocytes with comparable fertilization potentials as compared to those in Dgcr8flox/flox mice (Control). Gross morphology, histology, and weight of uteri of uDgcr8 KO mice were similar to those of control at 3-week-old stage. However, uterus become extremely thinner and shorter from 4-week-old stage onward. Histological examination showed significant reduction in gland numbers and stromal area from 4-week-old stage. Interestingly, this phenotype is reflected by significant increase of PR expression in the uteri of 4-week-old mice. In addition, stromal cell proliferation of uDgcr8 KO is severely impaired. BrdU incorporation experiments showed that while epithelial cells undergo proliferation by E2 treatment, stromal cells do not incorporate BrdU under the uterine conditions provided with E2+P4. Collectively, these results conclude that microRNAs are essential for uterine stromal cell proliferation in mice.

Agastache rugosa, a member of the mint family (Labiatae), is a perennial herb widely distributed in East Asian countries. It is used in traditional medicine for the treatment of cholera, vomiting, and miasma. This study assessed the genetic diversity and population structures on 65 accessions of Korean mint A. rugosa germplasm based on inter simple sequence repeat (ISSR) markers. The selected nine ISSR primers produced reproducible polymorphic banding patterns. In total, 126 bands were scored; 119 (94.4%) were polymorphic. The number of bands generated per primer varied from 7 to 18. A minimum of seven bands was generated by primer 874, while a maximum of 18 bands was generated by the primer 844. Six primers (815, 826, 835, 844, 868, and 874) generated 100% polymorphic bands. This was supported by other parameters such as total gene diversity (HT) values, which ranged from 0.112 to 0.330 with a mean of 0.218. The effective number of alleles (NE) ranged from 1.174 to 1.486 with a mean value of 1.351. Nei's genetic diversity (H) mean value was 0.218, and Shannon's information index (I) mean value was 0.343. The high values for total gene diversity, effective number of alleles, Nei's genetic diversity, and Shannon's information index indicated substantial variations within the population. Cluster analysis showed characteristic grouping, which is not in accordance with their geographical affiliation. The implications of the results of this study in developing a strategy for the conservation and breeding of A. rugosa and other medicinal plant germplasm are discussed.

This study deals with the design of a spring backboned soft arm, which will be employed for generation of human gesture as an effective means of Human‐Robot interaction. The special features of the proposed mechanism are the light weight and the flexibility of the whole mechanism by using a spring backbone. Thus, even in the case of collision with human, this device is able to absorb the impact structurally. The kinematics and the design for the soft arm are introduced. The performance of this mechanism was shown through experiment emulating several human gestures expressing human emotion and some service contents. Finally, this soft arm was implemented as the wing mechanism of a penguin robot.