• KOREAN JOURNAL OF CROP SCIENCE
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• v.45 no.3
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• pp.151-157
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• 2000

This study was carried out to isolate and identify the maysin and related flavonoid analogues in corn silks. Silks were covered with silk bag to prevent pollination and were sampled at 3-5 days after silking. The silks were filled with 100% MeOH and stored at $0^{\circ}C$ until analysis. The MeOH extracts of corn silks were filtered and concentrated at 35-4$0^{\circ}C$. The ${CH}_2$${Cl}_2$ was added on the concentrated aqueous solution to remove the chlorophyll and lipids. The Cis open column (25mm$\times$54 cm) was washed and activated with serial treatment of 500$m\ell$ of 100% MeOH(twice)longrightarrow75% MeOH longrightarrow50% MeOHlongrightarrow30% MeOHlongrightarrow100% $H_2$O(2 times). The concentrated aqueous solution was applied to the $C_{18}$ column and washed with $H_2O$ several times to remove the sugars and water soluble pigments. Neochlorogenic acid, chlorogenic acid and 4-caffeoylquinic acid were eluted with 10% MeOH, and rhamosyl isoorientin was eluted with 30% MeOH, but maysin was eluted with 50% MeOH from the $C_18$ open column. Collected fractions were analyzed with HPLC by using revers-phase Ultras-phere $C_{18}$ column (4.6$\times$250mm, 5$\mu\textrm{m}$) and $H_2$O (10% MeOH containing 0.1% $H_3$${PO}_4$)/MeOH (100% MeOH containing 0.1% H$_3$PO$_4$) linear gradient from 20% to 90% MeOH for 35 minutes, a flow rate of 1 $m\ell$/min and detection at 340nm. The selected fractions were concentrated and applied to the silicic acid column. Maysin was eluted with 500$m\ell$ of 100% ethyl acetate from the silicic acid column for the first purification, and the purity of collected fractions was about 75%, but the purity from the second purification with the Cis column (1/2 $\times$ 43") was greater than 95%. FAB-MS spectral data was obtained with VG7O-VSEQ VG analytical fast atom bombardment mass (UK). $^1$H-NMR and $^{13}$ C-NMR data were obtained with Bruker DPX 400 MHz NMR spectrometers (German) in DMSO-d$_{6}$ at 400 and 100 MHz, respectively.vely.

• Development and Reproduction
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• v.10 no.1
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• pp.1-7
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• 2006

Ovarian follicular atresia in mammals is finely regulated by gonadotropins and sex steroid hormones. It is well known that granulosa cell pyknosis is a common cytological feature of atretic follicles in the ovary. The present study hypothesized that granulosa cell pyknosis during follicular atresia might be related to apoptotic process and associated with caspase-3 activation. Healthy (normal) and atretic follicles were isolated from porcine ovaries based on macro-morphological criteria. Isolated follicles were either processed for histological observation or used for collection of granulosa cells by aspiration. Hoechst 33258 staining of the cells showed a significantly higher number of fragmented nuclei, a typical morphological feature of apoptotic cell, in granulosa cells from atretic follicles than those from healthy follicles. In addition, the rate of cell death was significantly higher in granulosa cells from atretic follicles than healthy follicles, as measured by flow-cytometric cell cycle analysis. In situ detection of apoptotic cells by TUNEL revealed that apoptosis was mostly restricted to granulosa cells in follicles. Theca cells were TUNEL-negative. Finally, it has been shown by caspase-3 activity assay that granulosa cells from atretic follicles retain a higher caspase-3 activity compared to healthy follicles. Taken together, it is suggested that granulosa cell degeneration during folliclar atresia occurs by caspase-3-dependent apoptotic fashion.

• Clinical and Experimental Pediatrics
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• v.49 no.4
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• pp.439-445
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• 2006

Purpose : ${\alpha}$-Galactosylceramide (${\alpha}$-GalCer)-stimulated human $V{\alpha}24$ natural killer T (NKT) cells exert antitumor activity against some leukemia in a CD1d dependent and TCR-mediated manner, but could not kill CD1d - negative neuroblastoma (NB) cells. There are few reports about the direct antitumor effect of highly secreted cytokines by these cells on activation. In this study, using a cell-free supernatant (SPN) collected from plate bound hCD1d/${\alpha}$ GalCer tetramers-stimulated NKT cells, we examined whether they could be helpful in the immunotherapeutic treatment of NB. Methods : Cells were cultured in IMDM. The cytokines produced by NKT cells were measured with Cytometric Bead Array (CBA) analysis. Cell viability was evaluated by calcein-AM fluorescence with digital image microscopy scanning (DIMSCAN). The percentage of specific apoptosis was calculated by flow cytometric detection of apoptosis using annexin V and 7-AAD. Results : The activated NKT cells secreted high levels of IL-2, INF-${\gamma}$, TNF-${\alpha}$. The SPN was significantly cytotoxic against four out of eight tested NB cell lines, through mainly apoptosis as evidenced by annexin-V staining and inhibition with the pretreatment of pancaspase blocker. This apoptosis was significantly inhibited when anti-TNF-${\alpha}$ and anti-IFN-${\gamma}$ neutralizing mAbs were used separately and it was completely abolished when the two mAbs were combined. Conclusion : IFN-${\gamma}$ and TNF-${\alpha}$ produced by NKT cells could exert synergistically direct antitumor activity through apoptosis on some NB cell lines.

• Journal of Korean Society of Environmental Engineers
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• v.27 no.6
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• pp.590-598
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• 2005

An adequate method to identify chromium separation, Cr(III) and Cr(VI), in water samples were studied by using High Performance Liquid Chromatography(HPLC) coupled with Inductively Coupled Plasma Mass Spectometer(ICP-MS) equipped with Dynamic Reaction Cell(DRC). The characteristic distribution of Cr(III) and Cr(VI) in the raw water taken at the six water intake stations in Seoul, was analyzed by the method developed by the authors. The chromium species separated by HPLC was isocratically conducted by using tetrabutylammonium phosphate monobasic(1.0 mM TBAP), ethylenediaminetetraacetic acid(0.6 mM EDTA) and 2% v/v methanol as the mobile phase. 5% v/v methanol was used as flushing solvent. A reactive ammonia($NH_3$) gas was used to eliminate the potential interference of $ArC^+$. Several Parameters such as solvent ratio, pH, flow rate and sample injection volume were optimized for the successful separation and reproducibility. Although it has been reported thai the separation sensitivity of Cr(III) is superior to that of Cr(VI), the authors observed Cr(VI) was more sensitive than Cr(III) when ammonia($NH_3$) gas was used as the reaction gas. It took less than 3 minutes to analyze chromium species with this method and the estimated detection limits were $0.061\;{\mu}g/L$ for Cr(III) and $0.052\;{\mu}g/L$, for Cr(VI). According to the results from the analysis on chromium species in the raw water of the six intake stations, the concentrations of Cr(III) ranged from 0.048 to $0.064\;{\mu}g/L$(ave. $0.054\;{\mu}g/L$) while that of Cr(VI) ranged from 0.014 to $0.023\;{\mu}g/L$(ave. $0.019\;{\mu}g/L$). Recovery ratio was very high($90.1{\sim}94.1%$). There were two or three times more Cr(III) than Cr(VI) in the raw water.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.1
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• pp.61-67
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• 2016

An HPLC analysis method was developed for standard determinations of chlorogenic acid, 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, and 4,5-di-O-caffeoylquinic acid as functional health materials in Ligularia fischeri extract. HPLC was performed on a $C_{18}$ Kromasil column ($4.6{\times}250mm$, $5{\mu}m$ column) with a gradient elution of 0.1% (v/v) trifluoroacetic acid and acetonitrile at a flow rate of 1.0 mL/min at $30^{\circ}C$. The analytes were detected at 330 nm. The HPLC method was validated in accordance with the International Conference on Harmonization guideline of analytical procedures with respect to specificity, precision, accuracy, and linearity. The limits of detection and quantitation for the four compounds were 3.0~14.6 and $9.2{\sim}44.4{\mu}g/mL$, respectively. Calibration curves showed good linearity ($r^2$ > 0.999), and the precision of analysis was satisfied (less than 0.9%). Recoveries of quantified compounds ranged from 98.96 to 101.81%. This result indicates that the established HPLC method is very useful for the determination of marker compounds in Ligularia fischeri leaf extracts.

• Journal of the Korean Society of Radiology
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• v.7 no.6
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• pp.369-376
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• 2013

Prepared her for the early detection of the risk of cardiovascular disease prevention for the various attempts being made ill through a lot of research to the criteria of coronary artery calcification, but the figure is more than 100 the progress of the flow can be expected to be was. In this study, we quantify the correlation between body composition analysis, blood lipid levels of calcium and public asymptomatic even at low calcium levels in comparison with the existing studies by analyzing potential risk of cardiovascular disease will be represented on the were evaluated. Studies, the calcium scores in the body composition analysis "1-10", and when "11-100" when there was a significant correlation in BMI and WHR look more normal frequency range of the mean values in the normal range meomulreoteuna 21% relative HDL and TG in lipid profiles, no significant correlation look more normal frequency range of risk were derived from the mean values in the normal range, 21% to 40% of the relative risk were derived. meomulreoteuna. In addition, there is no correlation between BFM and BMR Also, average increases in the frequency of a higher standard was higher relative risk derived 31 to 93%, and geological correlation test TC above 20mg/dl atherosclerotic sclerosis seems to be the value of HDL to act to remove cholesterol from atheroma already looking at the same time to suppress the occurrence of atherosclerosis, increased LDL showed higher values occurs rapidly increasing. At this time, the relative risk of 43-50% have been identified. In other words, even when calcium levels are low, that is inherent in the incidence of cardiovascular disease was unknown.

• Journal of Chest Surgery
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• v.34 no.9
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• pp.653-661
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• 2001

Background: Retrograde cerebral perfusion(RCP) is one of the methods used for brain protection during aortic arch surgery. The author previously published the data, however, for the safety of it, there still remains many controversies. The author performed RCP and checked various parameters to clarify the possibility of early detection of cerebral injury. Material and Method: The author used pigs(Landrace species) weighing 25 to 30kg and performed RCP for 120 minutes. After weaning of cardiopulmonary bypass, we observed pigs for another 120 minutes. Rectal temperature, jugular venous oxygen saturation, central venous pressure were continuously monitored, and the hemodynamic values, histological changes, and serum levels of neuron-specific enolose(NSE) and S100$\beta$ protein were checked. Central venous pressure during RCP was maintained in the range of 20 to 25 mmHg. Result: Flow rates(ml/min) during RCP were 224.3$\pm$87.5(20min), 227.1$\pm$111.0(40min), 221.4$\pm$119.5(60min), 230.0$\pm$136.5(80min), 234.3$\pm$146.1(100min), and 184.3$\pm$50.5(120min). Serum levels of NSE did not increase after retrograde cerebral perfusion. Serum levels of S100$\beta$ protein(ng/ml) were 0.12$\pm$0.07(induction of anesthesia), 0.12$\pm$0.07(soon after CPB), 0.19$\pm$0.12(20min after CPB), 0.25$\pm$0.06(RCP 20min), 0.29$\pm$0.08(RCP 40min), 0.41$\pm$0.05(60min), 0.49$\pm$0.03(RCP 80min), 0.51$\pm$0.10(RCP 100min), 0.46$\pm$0.11(RCP 120min), 0.52$\pm$0.15(CPBoff 60min), 0.62$\pm$0.15(60min after rewarming), 0.76$\pm$0.17(CPBoff 30min), 0.81$\pm$0.20(CPBoff 60min), 0.84$\pm$0.23(CPBoff 90min) and 0.94$\pm$0.33(CPBoff 120min). The levels of S100$\beta$ after RCP were significantly higher than thosebefore RCP(p<0.05). The author could observe the mitochondrial swellings using transmission electron microscopy in neocortex, basal ganglia and hippocampus(CA1 region). Conclusion: The author observed the increase of serum S100$\beta$ after 120 minutes of RCP. The correlation between its level and brain injury is still unclear. The results should be reevaluated with longterm survival model also considering the confounding factors like cardiopulmonary bypass.

• International Journal of Vascular Biomedical Engineering
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• v.4 no.2
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• pp.19-24
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• 2006

Background: Pulse wave velocity (PWV), which is inversely related to the distensibility of an arterial wall, offers a simple and potentially useful approach for an evaluation of cardiovascular diseases. In spite of the clinical importance and widespread use of PWV, there exist no standard either for pulse sensors or for system requirements for accurate pulse wave measurement. Objective of this study was to assess the reproducibility of PWV values using a newly developed PWV measurement system in healthy subjects prior to a large-scale clinical study. Methods: System used for the study was the PP-1000 (Hanbyul Meditech Co., Korea), which provides regional PWV values based on the measurements of electrocardiography (ECG), phonocardiography (PCG), and pulse waves from four different sites of arteries (carotid, femoral, radial, and dorsalis pedis) simultaneously. Seventeen healthy male subjects with a mean age of 33 years (ranges 22 to 52 years) without any cardiovascular disease were participated for the experiment. Two observers (observer A and B) performed two consecutive measurements from the same subject in a random order. For an evaluation of system reproducibility, two analyses (within-observer and between-observer) were performed, and expressed in terms of mean difference ${\pm}2SD$, as described by Bland and Altman plots. Results: Mean and SD of PWVs for aorta, arm, and leg were $7.07{\pm}1.48m/sec,\;8.43{\pm}1.14m/sec,\;and\;8.09{\pm}0.98m/sec$ measured from observer A and $6.76{\pm}1.00m/sec,\;7.97{\pm}0.80m/sec,\;and\;\7.97{\pm}0.72m/sec$ from observer B, respectively. Between-observer differences ($mean{\pm}2SD$) for aorta, arm, and leg were $0.14{\pm\}0.62m/sec,\;0.18{\pm\}0.84m/sec,\;and\;0.07{\pm}0.86m/sec$, and the correlation coefficients were high especially 0.93 for aortic PWV. Within-observer differences ($mean{\pm}2SD$) for aorta, arm, and leg were $0.01{\pm}0.26m/sec,\;0.02{\pm}0.26m/sec,\;and\;0.08{\pm}0.32m/sec$ from observer A and $0.01{\pm}0.24m/sec,\;0.04{\pm}0.28m/sec,\;and\;0.01{\pm}0.20m/sec$ from observer B, respectively. All the measurements showed significantly high correlation coefficients ranges from 0.94 to 0.99. Conclusion: PWV measurement system used for the study offers comfortable and simple operation and provides accurate analysis results with high reproducibility. Since the reproducibility of the measurement is critical for the diagnosis in clinical use, it is necessary to provide an accurate algorithm for the detection of additional features such as flow wave, reflection wave, and dicrotic notch from a pulse waveform. This study will be extended for the comparison of PWV values from patients with various vascular risks for clinical application. Data acquired from the study could be used for the determination of the appropriate sample size for further studies relating various types of arteriosclerosis-related vascular disease.

• Tuberculosis and Respiratory Diseases
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• v.41 no.4
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• pp.354-362
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• 1994

Objectives and Methods : The presence of aneuploidy or high proliferative activity in cytologic specimens is considered as complementary for the diagnosis of malignancy. To evaluate the diagnostic usefulness of DNA ploidy and cell cycle analysis in lung cancer, we compared the diagnostic yielding rates of DNA ploidy test by brushing specimens using flow cytometry with bronchoscopic forceps biopsy and brushing cytology. Results : Of the seventy-six cases, 55 cases proved to have malignant diseases(squamous cell cancer: 27, adenocarcinoma: 7, large cell cancer: 1, undifferentiated: 4 and small cell cancer: 16). The incidence of aneuploidy in lung cancer patients was 32.7%(18/55), as opposed to no cases in benign disease. And the proportion of high proliferative activity(S+G2M>22%) in lung cancer patients was 42.9%(15/35), but none in benign diseases. In fifty-six of 75 cases(74.7%), cytology of brushing specimens and DNA analysis(either aneuploidy or high proliferative activity vs. diploidy and low proliferative activity) were in concordance. The sensitivity with only brushing cytology was 41.8%(23/55), but with the addition of DNA analysis, it was increased to 56.4%(31/55), without decreasing the specificity(100%). And there was a case whose clue for malignancy was absent except aneuploidy, and he was confirmed to have squamous cell cancer following open thoracotomy. There were no differences in the frequency of aneuploidy or high proliferative activity between histologic subtypes of bronchogenic malignancy. Conclusions : The diagnostic detection rate of lung cancer was improved with the addition of DNA ploidy and cell cycle analysis, and the presence of aneuploidy or high proliferative activity was a relatively specific indicator of malignant disease. It would be useful to test DNA ploidy and cell cycle analysis with brushing specimen for the diagnosis of bronchogenic malignancy particularly in patients whose biopsy specimen could not be obtainable.

• Tuberculosis and Respiratory Diseases
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• v.40 no.5
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• pp.540-547
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• 1993

Backgrounds: The measurement of volume of isoflow has been considered as a sensitive test for detecting small airway diseases showing normal pulmonary function in a routine pulmonary function test. To evaluate the functions of small airway among dust exposed workers, the changes of volume of isoflow were measured and its applicability of managing early stage pneumoconiosis patients was studied. Method: The subjects were 67 male, pneumoconiosis with small opacity and FEV1>80%, FEV1/FVC>75% in spirometry and the controls were 20 male, no dust-exposed office workers. The maximal epiratory volume curves after inhalation of indoor air and $He-O_2$ gas mixtures were measured and ${\Delta}V_{max50},\;{\Delta}V_{max75},\;V_{iso}V/VC$ between the dust exposed and control workers were compaired. Results: 1) There were no significant differences between two group in ${\Delta}V_{max50}$ and ${\Delta}V_{max75}$. But the ratio of $V_{iso}V/VC$ of the subjects was siginificantly higher than that of the control (p<0.01). This study confirms that $V_{iso}V/VC$ is a very useful index in early detection of small airway dysfunction. 2) The ratio of $V_{iso}V/VC$ of the subjects was signigicantly different between only smoker group and mixed group(smoker and nonsmoker). It suggestes that smoking is an important cousative factor of small airway dysfunction. 3) As the profusion of the chest X-ray increased, the rartio of $V_{iso}V/VC$ increased, but no significant difference of $V_{iso}V/VC$ was found between categories of pneumoconiosis. The categories of pneumoconiosis and small airway dysfunction may not be related. 4) No significant relationship was established between the duration of work and the ratio of $V_{iso}V/VC$. Conclusions : It is concluded that the measurement of $V_{iso}V/VC$ is useful to detect small airway dysfuction of early stage pnuemoconiosis patents with small opacities but showing normal pulmonary function in a routine pulmonary function test.