본 연구는 비정시 환자의 양안시 조건과 관련하여， 양안굴절검사를 시행할 경우 구 면 원주 굴절력과 원주 축의 변화들에 영향을 주는지 그리고 어떤 방법으로 영향을 주는지를 평가하기 위해 시행되었다. 기존의 연구에 따르면 양안굴절검사를 시행할 경우 대부분의 환자들에서 마이너스 굴절력의 감소와 축의 변화가 있다고 하였다.7 세에서 58 세 사이의 다양한 연령대의 115명의 환자들을 대상으로 통상적인 단안굴절 검사 후 경사각이 설정된 험프리 즉시 대비 (HIC) 검사를 실시하였다. 원거리 안편위 와 원주축의 편위 사이에 상관관계가 있었다 (F = 3.296, p = 0.012). 양안굴절검사에서 단위 조절량에 대한 조절성 폭주비 (AC/A 비)와 구면굴절력 (F = 1.627, p = 0.010), 원주 굴절력 (F = 1.739, p = 0.004) 및 원주 축의 편위 (F = 1.702, p = 0.005) 사이 에 유의 한 상 관관계가 있었다. 그러나 대상 환자에서 폭주 근점 (NPC) 과 구변굴절력 (F = 0.793, p = 0.667), 원주굴절력 (F = 0.783, p 二0.677) 및 원주축의 편위 (F = 1.448 p = 0.14이와는 어 떤 관련성도 없었다. 양안굴절검사에서 원주축의 내회선 편위 현상이 나타난 환자의 구변굴절력 (t = -3.452, p = 0.001) 과 원주굴절력 (t = -5.571, p 二0.000) 은 감소하였으며， 원주 축은 평균 5.73 :t 8.00。내회선하였다. 원주축의 외회선 편위가 발생한 환자의 원 주굴절력은 감소하였고 (t = -3.630, p = 0.000), 원주축은 평균 5.38 :t5.36。외회선 편위 하였으며， 원주축의 변화가 없는 경우 구면 원주굴절력의 변화도 없는 것으로 나타났 다 (t 二-1.621 ， p 二0.113). 원거리 안편위의 형태와 AC/A 비에 의해 구변 원주굴절력 값에 변화가 있으며， 원주축은 원거리 안편위 유형에 따라서 원주축의 회선방향에 특 정적인 차이가 있음을 알 수 있았다. 이상의 결과들은 단안굴절검사와 양안굴절검사 사이의 구변-원주굴절력 값과 원주축의 편위 값을 비교한 기존의 연구들의 결과들과 유사한 값을 보였다. 본 연구에서 나타난 결과들은 양안굴절검사에서 비정시 환자의 양안시 조건이 구면 원주굴절력의 변화와 원주축의 편위에 영향을 미치는 주요 인자 로서 역할을 할 수도 있다는 것을 나타낸다고 할 수 있다. 향후 좀 더 많은 환자를 대상으로 원거리 안편위 형태와 원주 축의 편위 사이의 관계에 대해 연구할 필요가 있을 것이다

Dendropanax morbifera Leveille (Araliaceae) is an endemic species growing in the south-western part of South Korea that has been used in folk medicine and health functional food. In this study, we investigated an extract of quercetin in Jeju D. morbifera by varying different parts (fruit, sprouts, leaves, sprigs, and branches), harvest times, and extraction solvents. In addition, we aimed to establish a simple and reliable HPLC/UV analytical method to determination of quercetin for the quality control and base line data of the Jeju D. morbifera extract as a health functional food ingredient. The analytical specificity was determined with retention time and photo diode array (PDA) spectrum by analyzing quercetin using HPLC and comparing the results to those of extracts. This analytical method for quercetin was validated for its limit of detection (LOD), limit of quantitation (LOQ), precision, and accuracy. A high linearity in the standard calibration curve was obtained, with a coefficient of determination (R2) of 0.9996. Also, the LOD and LOQ values were found to be 0.28 μg/mL and 0.85 μg/mL, respectively, and the recoveries of quantified compounds ranged from 97.91% to 104.10%. Furthermore, the relative standard deviation (RSD) values of data from the intra- and inter-day precision analyses were less than 1.36% and 3.65%, respectively. As a result, the highest quercetin content among the extracts of Jeju D. morbifera leaves was found to be 20.14 mg/g, which was extracted at harvest in May (cultivation period 10 years) with 60% EtOH. All in all, we believe that the results obtained would be helpful in the development of nutraceutics and natural medicines and for the quality control of D. morbifera.

Background : Heat stress induced from high temperature are known to crucially affecting on physiological properties and yield in Cnidium officinale. Methods and Results : The effect of foliar application of mixture including a urea, ascorbic acid and calcium chloride on high temperature injury of Cnidium officinale. Photosynthesis and leaf temperature in Cnidium officinale were investigated after cultivating for 24 hours at 35℃. Net photosyntheis rate, transpiration was measured at 1,000 μmol m-2 s-1 of photon flux density and leaf temperature was analyzed by thermal image. Net photosyntheis rate, stomatal conductance and transpiration rate in mixture traetment were 2 times of higher than in control. Water use efficience was not different significantly. Leaf temperature was lower in mixture treatment (25.3℃) than in control (29.0℃). Conclusion : This result show that foliar application of urea, ascorbic acid and calcium chloride was reducing a high temperature injury through a improving photosynthetical capacity and decreasing leaf temperature of Cnidium officinale.

Background : Management of air temperature are known to primarily affecting on physiological properties and yield in plant. Methods and Results : The effect of air temperature on characteristics of photosynthesis and chlorophyll fluorescence in Cnidium officinal were investigated using growth chamber after cultivating for 24 hours under controlled condition. Net photosyntheis rate, transpiration was measured at 1,000 μmol m-2 s-1 of photon flux density and chlorophyll fluorescence was analyzed by OJIP method. Net photosyntheis rate was highest in treatment of 25℃. Although transpiration rate was lowest, water use efficience was also in treatment of 25℃. Stomatal conductance was mainly influenced from ambient climatric factors such as vapor pressure deficit. As results of chlorophyll fluorescence by OJIP analysis, maximum quantum yield (Fv/Fm) of photosystem II (PSII), PIabs and the relative activities per reaction center such as ABS/RC, DIo/RC were not changed at air temperature. Therefore, elevated air temperatue during short term influence the dark reaction in photosystem through controlling a water use efficience and transpiration. Conclusion : This result show that 25℃ of air temperature may be a adequate temperature to improving the efficiency of photosynthesis in Cnidium officinale.

Hepatocytes and hepatic progenitors derived from human ES cells may be a useful source for clinical application. Therefore, identification and purification of these cell types would be following important issues. There are very few candidate surface markers that can be used to identify and purify hepatic progenitor cells. In addition, indocyanine-green can be uptaken by mature hepatocytes, but cannot be applied for fluorescence activated cell sorting (FACS) due to its long emission wavelength. In the present study, we tested EpCAM as a potential marker for magnetic-activated cell sorting (MACS) of hepatic progenitors and also modified indocyanine-green into fluorescent indomonocarbocyanine for FACS-mediated sorting of mature hepatocytes after differentiation of human ES cells. Hepatic progenitor cells were sorted by MACS after incubation with anti-human EpCAM antibodies. After the final differentiation, the differentiated cells and mouse primary hepatocytes (control group) were incubated with indomonocarbocyanine and were sorted by FACS. MACS and immunocytochemistry data showed that approximately 45% of differentiated cells were EpCAM-positive cells. EpCAM-positive cells expressed α-fetoprotein, FOXa2, HnF4a, and CK18. Differentiation efficiency into albumin-positive cells was significantly higher in EpCAM-positive cells, compared to EpCAM-negative cells. Importantly, indomonocarbocyanine successfully stained cells that expressed ALB. Furthermore, FACS analysis data showed that the purity of hepatocytes that expressed albumin was significantly increased after purification of indomonocarbocyanine-positive cells. Our data demonstrated that human ES cell-derived hepatic progenitors can be efficiently isolated by MACS using EpCAM antibody. In addition, we also showed that indomonocarbocyanine can be successfully used to identify and purify mature hepatocytes using FACS.

Highly homogeneous and functional stem cell-derived hepatocyte-like cells (HLCs) are considered a promising option in the treatment of liver disease and the development of effective in vitro toxicity screening tool. However, the purity of cells and expression and/or activity of drug metabolizing enzymes in stem cell-derived HLCs are usually too low to be useful for clinical or in vitro applications. Here, we describe a highly optimized differentiation protocol, which produces more than 90% albumin-positive HLCs with no purification process. In addition, we show that hepatic enzyme gene expressions and activities were significantly improved by generating three-dimensional (3D) spheroidal aggregate of HLCs. The 3D differentiation method increased expressions of nuclear receptors that regulate the proper expression of key hepatic enzymes. Furthermore, a significantly increased hepatic functions such as albumin and urea secretion were observed in 3D hepatic spheroids and HLCs in the spheroid exhibited morphological and ultrastructural features of normal hepatocytes. Importantly, we show that repeated exposures to xenobiotics facilitated the functional maturation of HLC, as confirmed by increased expression of genes for drug metabolizing enzymes and transcription factors. In conclusion, the 3D culture system with repeated exposures to xenobiotics may be a new strategy for enhancing hepatic maturation of stem cell-derived HLCs as a cell source for in vitro high-throughput hepatotoxicity models.

Hepatocyte-like cells (HLCs) derived from human pluripotent stem cells have received extensive attention in the development of drug screening and toxicity testing. However, it has been reported that stem cell-derived HLCs showed hepatic functions that were too limited to be of use in drug screening and toxicity testing, possibly due to the lack of sufficient intercellular communication under conventional two-dimensional (2D) culture conditions. Therefore, a 3D differentiation system may overcome the in vitro limitation of 2D culture to produce stem cell-derived hepatocytes with mature metabolic functions. In this study, the feasibility of using a silicone-based spherofilm, specifically designed to produce spherical cell clusters, to generate uniformly sized 3D hepatic spheroids from hESCs was investigated. Hepatic spheroids generated on the spherofilm showed more homogenous size and shape than those generated in conventional low-attachment suspension culture dishes. Results of immunohistochemical analysis showed that expression of the mature hepatic marker albumin (ALB) increased over time during the hepatic maturation process. Furthermore, the 3D culture system mimicked the in vivo 3D microenvironment. Laminin, which is an important component of hepatic ECM, was expressed in hepatic spheroids. The results of immunohistochemical analysis indicated that the 3D culture environment is capable of generating an in vivo-like microenvironment. In addition, quantitative PCR analysis showed that the mature hepatic marker ALB and cytochrome P450 (CYP) enzymes CYP3A4 and CYP3A7 were expressed at higher levels in 3D culture than in 2D culture. This indicates that the 3D culture system is suitable for hepatic maturation and that our size-controlled 3D culture conditions might accelerate hepatic function. These results suggest that 3D hepatic spheroids significantly enhance metabolic maturation of hepatocytes derived from hESCs

Chronic gamma irradiation can be used an alternative mutation breeding methods for induction of many useful mutants. Seedlings of purple-colored wheat plants were irradiated with wide range doses of chronic gamma-rays (20, 25, 30, 40, 50, 70, 100, 125, 150, 200, 250, 300 Gy) during 6 weeks at gamma-phytotron in the Korea Atomic Energy Research Institute, respectively. To identify the biological responses purple-colored wheat, we examined the plant height, chlorophyll, carotenoid and total anthocyanin contents in leaf. Plant growth, chlorophyll and carotenoid contents in leaf were decreased when the dose rate increased. Anthocyanin contents were increased with the increase of the radiation dose until 50 Gy treatment. To confirm the real contents of anthocyanin, we also investigated cyanidin-3-glucoside in purple-colored wheat leaf by using UPLC analysis. These results indicate that anthocyanin accumuation was observed under chronic gamma irradiation.

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.

Compared to wide ranges of genetic variation of natural populations, very limited Miscanthus cultivar has been released. This study was the first report on the development of Miscanthus cultivar by means of radiation breeding. Seeds of M. sinensis were initially exposed to gamma rays of 250 Gy for 24 h, generated from a 60Co gamma-irradiator. The irradiated seeds were sown and then the highly tiller-producing mutants were selected for this study. Biomass-related parameters including tiller number, plant height, stem diameter, and leaf number were measured. Ploidy level and internal transcribed spacer (ITS) were investigated to characterize the mutants compared to wild type (WT) Miscanthus. Plant height and tiller number were negatively related, where multi-tillering mutants were relatively short after 4 month growth. However stem diameter and leaf number were greater in mutants. All the materials used in this study were diploid, implying that the mutants with greater tiller numbers and stem diameter were not likely related to polyploidization. Based on the sequence of ITS regions, the mutants demonstrated base changes from the gamma irradiation where G+C content (%) was decreased in the ITS1, but increased in ITS2 when compared to WT sequence. ITS2 region was more variable than in ITS1 in the mutants, which collectively allows identification of the mutants from WT. Those mutants having enhanced tillers and allelic variations might be used as breeding materials for enhanced biomass-producing Miscanthus cultivars.

Cryopreservation has been known as an efficient method for long-term preservation of clonally propagated plants, and several cryopreservation methods have been developed. Among them, a droplet-vitrification method for potato using axillary shoot tips in vitro has been established previously. In this study, we have optimized the procedure in which explants were submitted to a step-wise pre-culture in liquid sucrose-enriched medium (0.3 and 0.7 M for 7 and 17 h, respectively). The pre-cultured explants were dehydrated with PVS3 (w/v, 50% glycerol + 50% sucrose) for 90 min or modified PVS2 vitrification solution (w/v, 37.5% glycerol + 15% DMSO + 15.0% ethylene glycol + 22.5% sucrose) for 30 min. This two dehydration solutions produced post-cryopreservation regeneration percentages of 57.2% and 80.9%, respectively. We also compared a new post-culture medium (0.1 mg L ･ -1 GA3, 0.1 mg L ･ -1 kinetin) with the conventional one (0.15 mg L ･ -1 IAA, 0.2 mg L ･ -1 zeatin, 0.05 mg L ･ -1 GA3); the shooting initiation rates were 80.9% and 43.5%, respectively. The results suggest that the modified droplet-vitrification protocol described in this study is more effective, easier to implement, and more economical than the droplet-vitrification protocols currently used for potato.