• Development and Reproduction
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• v.12 no.1
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• pp.1-8
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• 2008

Specific protocols to increase the differentiation of neuronal cells from embryonic stem (ES) cells have been well established, such as retinoic acid induction and lineage selection of neuronal cells. For the neuropathological studies, ES-derived neurons (ES neurons) must show normal physiological characteristics related to cell death and survival and should be maintained in vitro for a sufficient time to show insults-specific cell death without spontaneous death. When mouse ES cells were plated onto astrocytes monolayer after retinoic acid induction, most ES cells differentiated into neuronal cells, which were confirmed by the presence of specific neuronal markers, and the cultures were viable for at least four weeks. When these cultures were examined for vulnerability to glutamate excitotoxicity, ES neurons were vulnerable to excitotoxic insults mediated by agonist-specific receptors. The vulnerability to excitotoxic death increased with developmental age of ES neurons in vitro. Specific receptors for Neurotrophin and GDNF family ligands were present in ES neurons. GDNF and NT-3 could modulate the survival and excitotoxic vulnerability of ES neurons. The vulnerability and resistance to toxic insults, which are essential requirements of model culture systems for neuropathological studies, make ES neurons to a useful model culture system. Especially ES cell are highly amenable to genetic modification unlikely to primary neuronal cells, which will give us a chance to answer more complicated neurophysiological questions. Recently there was an outstanding attempt to explore the cellular toxicity using human ES cells (Schrattenholz & Klemm, 2007) and it suggested that ES cells could be a new model system for neurophysiological studies soon and go further a large-scale screening system for pharmacological compounds in the future.

• Development and Reproduction
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• v.14 no.2
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• pp.99-105
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• 2010

The seminiferous epithelium, with its division into 12 spermatogenic stages in the mouse, is a very complex tissue. Glutathione peroxidase (GPx) is a representative antioxidant enzyme that is capable of reducing organic hydroperoxides to their corresponding hydroxyl compounds utilizing glutathione and is related to the mammalian spermatogenesis. In this study, a real-time PCR was performed in the stage-specific seminiferous tubules of mouse testes excised by a laser capture microdissection (LCM) in order to quantitate the expression levels of a series of GPx genes including cytosolic GPx (cGPx), gastrointestinal GPx (GI-GPx), plasma GPx (pGPx), and phospholipid hydroperoxide GPx (PHGPx). Frozen sections (10 ${\mu}m$) were obtained from normal adult mouse testes. LCM was used to capture all the cells that were grouped into stages I-V, VII-VIII, and IX-XI in cross-sections of seminiferous tubules. The expression level of PHGPx mRNA was remarkably higher than those of other GPx mRNAs in mouse testes. During spermatogenesis, the expressions of GI-GPx, pGPx, and PHGPx mRNAs were highest on stages VII-VIII, began to decrease after stage XI, and showed a lowest level on stage I-V. However, the expressions of cGPx mRNA were highest on stages VII-VIII, and showed a lowest level on stage XI-XI. These findings indicate that GPx genes are expressed differentially on mouse spermatogenesis and also LCM can be an useful tool in cellular quantitative analysis of testes.

• Microbiology and Biotechnology Letters
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• v.28 no.2
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• pp.97-104
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• 2000

Screening of Biologically Active Essential Oils from Ligusticum tenuissimum. Kim, Min-Hae, Young-Gil Kim, Jin-Ha Lee, Keo-Pyo Hong, Jung-Ki Hong, Young-Joon Kong, and Hyeon-Yong Lee*. Division of Food and Biotechnology, Kangwon National University, Chunchon 200-701, Korea, 1 Regional Crop Development Station, Kangwon Agricultural Research & Extension Services, Chunchon 200-150, Korea-The biological activities of the crude essential oils from Ligusticum tenuissimum and the control(phthalic anhydride) were compared. About 60% of the growth of MCF7, A549, and Rep3B cells were inhibited by adding 1.0 mg/ml of the crude essential oils and below 40% was observed by the control. Cytotoxicity on human normal lung cell(IMR90) was scored as 34.4% for the crude oil and 26.4% for control, respectively. It was found that the crude essential oils were more effective than the control in anti mutagenecity tested by both Rec-assay and CRG V79 cells. The growth of human T-cell(Jurkat) was enhanced up to 1.21 times by adding the crude essential oil compared with the control. 50% of a-glucosidase activity was inhibited by both the crude essential oil and the control. ACE activities were inhibited 80.1 % and 65.3% by adding 1.0 mg/ml of the crude oil and the control, respectively. The higher enhancement of glutathione-S-transferase activity was observed in the crude oil than those in the control: 301 % v.s 234% at 1.0 mg/ml of the treatment. Thrombolytic activity was measured as 42.9% and 28.6% for the crude oil and the standard, respectively. The effect of the oil on the nerve cells PCI2, was observed as follows: the neurite of PCl2 cells was lengthened up to 255 /-lm longer than 205 /-lm of control. The number of neurite-bearing cells were about two times higher than control. The survival ratio of the crude essential oil was also increased up to 56.4% which was about two fold higher than in control.

• Journal of Pharmaceutical Investigation
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• v.34 no.1
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• pp.49-55
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• 2004

To develop a aceclofenac soft capsule, four preparations with various solubilizers were prepared and their dissolution test was carried out. Among four preparations tested, a preparation with ethanolamine was selected as a formula of aceclofenac soft capsule (Clanza $S^{TM}$), since it showed the fastεst dissolution rate. Bioequivalence of aceclofenac tablet, $Airtal^{TM}$ (Dae-Woong Pharmaceutical Co., Ltd.) and aceclofenac soft capsule, Clanza $S^{TM}$ (Korea United Pharmaceutical Co., Ltd.) was evaluated according to the guideline of KA Fourteen normal male volunteers (age 20 - 25 years old) were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After oral administration of one tablet or capsule containing 100 mg of aceclofenac, blood was taken at predetermined time intervals and the concentration of aceclofenac in plasma was determined with an HPLC method under UV detector The pharmacokinetic parameters ($C_{max}$ and $AUC_t$) were calculated and ANOVA was utilized for the statistical analysis of parameters using logarithmetically transformed $AUC_t$, $C_{max}$ and $T_{max}$. The results showed that the differences in $AUC_t$, $C_{max}$ and $T_{max}$ between Aral tablet and Clanza soft capsule were 2.89%, 0.18% and 43.0%, respectively. There were no sequence effects between two formulations in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log(0.8) to log(15) (e.g. log(0.81) -log(1.23) ad log(0.89) -log(1.4)) fo $AUC_t$ and $C_{max}$, respectively. Thus, the criteria of the KFDA guidelines for the equivalence was satisfied, indicating that Clanza $S^{TM}$ soft capsule is bioequivalent to$Airtal^{TM}$ tablet.

• Korean Journal of Agricultural and Forest Meteorology
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• v.4 no.4
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• pp.224-236
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• 2002

A long-term growth simulation was performed at 496 land units in the western coastal plains (WCP) of North Korea to test the potential adaptability of each land unit for growing South Korean rice cultivars. The land units for rice cultivation (CZU), each of them represented by a geographically referenced 5 by 5 km grid tell, were identified by analyzing satellite remote sensing data. Surfaces of monthly climatic normals for daily maximum and minimum temperature, precipitation number of rain days and solar radiation were generated at a 1 by 1 km interval by spatial statistical methods using observed data at 51 synoptic weather stations in North and South Korea during 1981-2000. Grid cells felling within a same CZU and, at the same time, corresponding to the satellite data- identified rice growing pixels were extracted and aggregated to make a spatially explicit climatic normals relevant to the rice growing area of the CZU. Daily weather dataset for 30 years was randomly generated from the monthly climatic normals of each CZU. Growth and development parameters of CERES-rice model suitable for 11 major South Korean cultivars were derived from long-term field observations. Eight treatments comprised of 2 transplanting dates $\times$ 2 cropping systems $\times$ 2 irrigation methods were assigned to each cultivar. Each treatment was simulated with the randomly generated 30 years' daily weather data (from planting to physiological maturity) for 496 land units in WCP to simulate the growth and yield responses to the interannual climate variation. The same model was run with the input data from the 3 major crop experiment stations in South Korea to obtain a 30 year normal performance of each cultivar, which was used as a "reference" for comparison. Results were analyzed with respect to spatial and temporal variation in yield and maturity, and used to evaluate the suitability of each land unit for growing a specific South Korean cultivar. The results may be utilized as decision aids for agrotechnology transfer to North Korea, for example, germplasm evaluation, resource allocation and crop calendar preparation.

• Horticultural Science & Technology
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• v.32 no.2
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• pp.157-164
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• 2014

Objective of this research was to develop root media containing expanded rice hull (ERH) and carbonized rice hull (CRH). To achieve this, the physico chemical properties of two materials were analysed and blended with peatmoss (PM) or coir dust (CD) with various ratio. Based on the physical properties of the blended materials, 4 root media were selected for future experiment. After the analysis of pH and EC of the selected root media, the kinds and amount of pre-planting nutrient charge fertilizers (PNCF) incorporated into each root medium were varied, and then, final chemical properties of the root media were analysed. The total porosity (TP), container capacity (CC), and air-filled porosity (AFP) were 81.3%. 39.9%, and 41.4% in ERH and 77.6%, 64.1%, and 13.5% in CRH, respectively. The percentage of easily available water (EAW, from CC to 4.90 kPa tension) and buffering water (BW, 4.91-9.81 kPa tension) were 11.37% and 5.27%, in ERH and 17.26% and 14.28% in CRH, respectively. The pH of ERH was 7.1, but it was extremely high in CRH such as 11.2. The EC and CEC were $1.31dS{\cdot}m^{-1}$ and $12.1meq{\cdot}100g^{-1}$ in ERH and $6.53dS{\cdot}m^{-1}$ and CEC 7.79 $meq{\cdot}100g^{-1}$ in CRH, respectively. The ranges of TP, CC and AFP in 4 selected media (PM + ERH, 6:4, v/v; CD + ERH, 8:2; PM + CRH, 7:3; CD+CRH 6:4) were 89.2-90.3%, 67.3-81.8%, and 8.3-21.9%, respectively. The pHs and ECs in root media containing peatmoss such as PM + ERH (6:4) and PM + CRH (7:3) were 4.0-4.3 and $0.33-0.365dS{\cdot}m^{-1}$, whereas those of CD + CRH were 7.4-7.9 and $1.282dS{\cdot}m^{-1}$. The pHs and ECs, however, analysed before and after the incorporation of PNCF in each root medium were not significant different. This result indicated that the incorporated fertilizers in PNCF to adjust medium pH did not dissolve enough to influence medium pH, but it is very normal in root media containing dolomitic lime and sulfur powder in adjusting pH. The Information obtained in this study may facilitate an effective formulation of root media containing rice hulls.

• Clinical and Experimental Pediatrics
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• v.48 no.7
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• pp.772-778
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• 2005

Purpose : Kawasaki disease(KD) is a systemic panvasculitis that causes coronary artery lesions. KD is accompanied by immunoregulatory abnormalities. Nitric oxide(NO) can induce relaxation of blood vessels by activating guanylate cyclase in smooth muscle cells and high levels of NO may result in coronary artery lesions. We investigated tumor necrosis factor$(TNF)-{\alpha}$ and NO production before and after intravenous immunoglobulin(IVIG) therapy to study the roles of NO and $TNF-{\alpha}$ in KD with coronary artery lesions. Methods : Serum levels of NO and $TNF-{\alpha}$ were measured in 24 patients with KD(group I, eight patients with normal coronary artery; group II, 16 patients with coronary artery lesions) and 23 controls(group III, 13 afebrile controls; group IV, 10 febrile controls). Blood samples from each subject were drawn before and after IVIG therapy and in the convalescent stage. Serum concentrations of NO and $TNF-{\alpha}$ were measured by enzyme linked immuno sorbent assay. Results : The NO levels before IVIG therapy were significantly higher in group II than in group I, group III and group IV. After IVIG therapy the levels of NO were significantly higher in group I and group II than in group III. The $TNF-{\alpha}$ levels before IVIG therapy were significantly higher in group I and group II than in group III. The serum $TNF-{\alpha}$ and NO levels were higher before IVIG therapy and decreased through the convalescent stage in KD patients. In the acute stage of KD patients with coronary artery lesions, serum NO levels significantly correlated with white blood cells (r=0.43, P<0.05). Conclusion : The serum concentration levels of $TNF-{\alpha}$ and NO were abnormally high in KD patients and NO concentrations were statistically higher in the KD patients with coronary artery abnormalities than those without coronary abnormality during the early stage of the KD. These results suggest NO may be involved in the development of coronary artery lesions.

• Clinical and Experimental Pediatrics
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• v.48 no.6
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• pp.624-633
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• 2005

Purpose : This study analyzed the expression of HLA-DR and DQ genotypes and anti-thyroid autoantibodies[anti-thyroid peroxidase(TPO) and anti-thyroglobulin(TG) antibodies] in Korean patients with type 1 diabetes(T1DM) to investigate the susceptible HLA alleles to T1DM in Korea and the prevalence of thyroid autoantibodies and their significance for the development of thyroid disorders. Methods : A total of 59 Korean patients with type 1 diabetes[26 males, median age 13.7 years(range 5.7-29.9 years), diabetes duration 7.6 years(-1.7-22.5 years)] were enrolled in this study, and 200 healthy Koreans without a family history of diabetes were selected as a normal control for the comparison of HLA genotypes. Seventeen patients with anti-TPO or anti-TG were followed [median duration 3.96 years(1 day-10.7 years)] with measurement of anti-TPO, anti-TG, $T_3$, $T_4$ or free $T_4$, TSH levels and physical examinations. HLA-DR and DQ genotyping were done by PCR-SSO, PCR-SSCP, PCR-RFLP and PCR-SSP methods. Results : HLA analysis showed higher frequencies of HLA-DRB1*0301, *090102 and DQB1*0201, *030302 alleles, DRB1*0301/*090102, *090102/*090102 and DQB1 *0201/*030302, *030302/*030302, *0201/ *0302 genotypes in T1DM patients compared to controls(Pc<0.05). Fifteen(25.4 percent) had anti-TPO antibody, 12(20.3 percent) had anti-TG, 17(28.8 percent) had either autoantibody and 10(16.9 percent) had both autoantibodies. No clinical or subclinical hypothyroidism developed during follow-up after the first detection of anti-thyroid autoantibody. There was no significant correlation between thyroid autoimmunity and gender, onset age of T1DM, and diabetes duration, respectively(P>0.05). Conclusion : We thought this unique HLA-DR, DQ allele distribution might be an important factor for the low incidence of T1DM in Korea. And a high prevalence of thyroid autoantibodies in these populations suggests examinations of thyroid antibodies should be performed regularly. Optimal age for the initial screening and the frequency of re-screening for associated thyroid autoimmune diseases in T1DM remains to be determined through prospective follow-up.

• Clinical and Experimental Pediatrics
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• v.48 no.7
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• pp.745-752
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• 2005

Purpose : Adolescent obesity is known to be associated with complications such as hypertension, coronary artery disease and insulin resistance. We measured the common carotid artery by ultrasound as a predictor of atherosclerosis, and investigated the relationship between carotid artery parameters and serum cytokines in obese adolescents. Methods : Twenty-nine obese adolescents(16-17 years old, obesity index>130 percent) and twentyseven normal controls were included. Obesity index and body mass index were calculated from their height and weight. Skin fold thickness was measured at the triceps, and fat mass and fat distribution by bioelectrical impedence analysis. Blood pressure was measured at resting state and serum lipid, insulin, and homocysteine levels after a 12-hour fasting period. Intimal wall thickness, systolic and diastolic diameters of the common carotid artery were measured by ultrasound, and compliance and distensibility calculated by equation. Results : Systolic and diastolic diameters of the carotid artery significantly correlated with arm circumference, body mass index, fat distribution and fat mass. The higher systolic blood pressure was, the larger systolic and diastolic diameter. The higher diastolic blood pressure was, the larger carotid intimal thickness. Insulin levels had positive correlations with systolic, diastolic diameters and serum homocysteine level with intimal thickness. Conclusion : The carotid artery diameter significantly increased with the degree of obesity and blood pressure. The carotid intimal wall thickness significantly correlated with diastolic blood pressure. Measurement of carotid artery thickness, insulin and homocysteine levels might be useful to predict the development of coronary artery disease.

• Journal of Yeungnam Medical Science
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• v.9 no.2
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• pp.359-381
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• 1992

This study was designed to investigate the effect of diazepam on the spontaneous contraction and oxytocin induced contraction of the isolated rat uterus. Female rat(Sprague-Dawley) pretreated with oophorectomy and 4 days administration of estrogen, weighing about 200 g, was sacrificed by cervical dislocation, and the uteruses were isolated. A longitudinal muscle strip was placed in temperature controlled($37^{\circ}C$) muscle chamber containing Locke's solution and myographied isometrically. Diazepam inhibited the spontaneous contraction and oxytocin induced contraction of the isolated rat uterus in a concentration-dependent manner. GABA, muscimol, a GABA A receptor agonist, bicuculline, a competitive GAGA A receptor antagonist, picrotoxin, a non competitive GABA A receptor antagonist, baclofen, a GABA B receptor agonist, and delta-aminovaleric acid, a GABA B receptor antagonist, did not affect on the spontaneous and oxytocin induced contraction of the isolated rat uterus. The inhibitory actions of diazepam on the spontaneous and oxytocin induced contraction were not affected by all the GABA receptor agonists and antagonists, but exceptionally potentiated by bicuculline. This potentiation-effect by bicuculline was not antagonized by muscimol. In normal calcium PSS, addition of calcium restored the spontaneous contraction preinhibited by diazepam and recovered the contractile of oxytocin preinhibited by diazepam. A23187, a calcium inophore, enhanced the restoration of both the spontaneous and oxytocin induced contraction by addition of calcium. In calcium-free PSS, diazepam suppressed the restoration of spontaneous motility by addition of calcium but allowed the recovery of spontaneous motility to a considerable extent. Diazepam could not inhibit some development of contractility by oxytocin in calcium-free PSS, but inhibited the increase in contractility by subsequent addition of calcium. These results suggest that the inhibitory action of diazepam on the rat uterine motility does not depend on or related to GABA receptors and that diazepam inhibits the extracellular calcium influx to suppress the spontaneous and oxytocin induced contractilities.