• Title/Summary/Keyword: Fluorescence intensity

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The Expression of Adhesion Molecules on Alveolar Macrophages and Lymphocytes and Soluble ICAM-1 Level in Serum and Bronchoalveolar Lavge(BAL) Fluid of Patients with Diffuse Interstitial Lung Diseases(DILD) (간질성 폐질환환자들의 기관지 폐포세척액내 폐포 대식세포와 임파구의 접착분자 발현 및 Soluble ICAM-1 농도에 관한 연구)

  • Kim, Dong-Soon;Choi, Kang-Hyun;Yeom, Ho-Kee;Park, Myung-Jae;Lim, Chai-Man;Koh, Yoon-Suck;Kim, Woo-Sung;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.4
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    • pp.569-583
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    • 1995
  • Background: The expression of the adhesion molecules on the cell surface is important in the movement of cells and the modulation of immune response. DILD starts as an alveolitis and progresses to pulmonary fibrosis. So adhesion molecules in these patients is expected to be increased. There are several reports about adhesion molecules in DILD in terms of the percentage of positive cells in immuno-stain, in which the interpretation is subjective and the data were variable. Methods: So we measured the relative median fluorescence intensity(RMFI) which is the ratio of the FI emitted by bound primary monoclonal antibody to FI emitted by isotypic control antibody of the cells in BALF of 28 patients with DILD(IPF:10, collagen disease:7, sarcoidosis:9, hypersensitivity pneumonitis:2) and 9 healthy control. Results: RMFI of the ICAM-1 on AM($3.30{\pm}1.16$) and lymphocyte($5.39{\pm}.70$) of DILD were increased significantly than normal control($0.93{\pm}0.18$, $1.06{\pm}0.21$, respectively, p=0.001, P=0.003). RMFI of the CD18 on lymphocyte was also higher($24.9{\pm}14.9$) than normal($4.59{\pm}3.77$, p=0.0023). And there was a correlation between RMFI of ICAM on AM and the % of AM(r=-0.66, p=0.0001) and lymphocyte(r=0.447, p=0.0116) in BALF. Also RMFI of ICAM on lymphocyte had a significant (r=0.593, p=0.075) correlation with the % of IL-2R(+) lymphocyte in BALF. The soluble ICAM(sICAM) in serum was also significantly elevated in DILD($499.7{\pm}222.2\;ng/ml$) compred to normal($199.0{\pm}38.9$) (p=0.00097) and sICAM in BAL fluid was also significantly higher than normal control group($41.8{\pm}23.0\;ng/ml$ vs $20.1{\pm}13.6\;ng/ml$). There was a Significant correlation between sICAM level in serum and the expression of ICAM-l on AM(r=0.554, p=0.0259).Conclusion: These data suggest that in DILD the expression of adhesion molecules is increased in the AM and BAL lymphocytes with elevated serum sICAM, and these parameter may be useful in determining disease activity.

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Characterization of Iron Oxides in Soils of Cheju Island by Mössbauer Spectroscopy and Chemical Techniques (Mössbauer 분광법(分光法)과 선택적(選擇的) 추출방법(抽出方法)에 의한 제주도(濟州道) 토양(土壤)에서의 산화철(酸化鐵)의 특성(特性) 구명(究明))

  • Kang, Dong-Woo;Kim, Doo-Chul;Ko, Jeong-Dae;Hong, Sung-Rak;Song, Kwan-Cheol
    • Korean Journal of Soil Science and Fertilizer
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    • v.30 no.1
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    • pp.3-15
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    • 1997
  • Iron oxide compounds in 8 selected Cheju Island soil samples have been analized by X-ray fluorescence spectrometer(XRF), X-ray diffractometry(XRD), selected chemical techniques, and $M{\ddot{o}}ssbauer$ spectroscopy. The result of this analysis by XRF shows that the rate of quantity of $Fe_2O_3$ in 8 soil samples was from 8.03wt.%(Daejeong paddy soil) to 18.21wt.%(Songag soils). Songag, Heugag and Gueom soils were detected to have lower peaks of intensity of hematite by XRD. In addition, these soils were not detected to have hematite and goethite peaks. Ferrihydrite, which is a short-range-order mineral commonly present in volcanic ash soil, was not detected by XRD due to low concentration and/or poor cristallinity. Ferrihydrite contents estimated from Feo values were 8.8~35.2g/kg for volcanic ash soils and 0.85g/kg for the Daejeong soil. Most of the soil samples represented by the paramagnetic $Fe^{3+}$ doublet obtained from $M{\ddot{o}}ssbauer$ spectra at room temperature and 18K were considered to arise from the presence of ferrihydrite, superparamagnetic goethite, and silicate minerals. Also the paramagnetic $Fe^{2+}$ doublets are attributable to primary minerals such as olivine, illite, chlorite, augite, biotite, and hornblende. Goethite and hematite were identified as the dominant crystalline iron oxides in these soils from $M{\ddot{o}}ssbauer$ spectra obtained at room temperature and 18K. All the soil samples exhibited strong superparamagnetic relaxation. Collapse of the $M{\ddot{o}}ssbauer$ magnetic hyperfine splitting at room temperature was due to the small size(${\sim}180{\AA}$) of the oxide particles and/or Al-subsituted goethite.

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Light Quality and Photoperiod Affect Growth of Sowthistle (Ixeris dentata Nakai) in a Closed-type Plant Production System (밀폐형 식물생산시스템에서 광질과 광주기에 따른 씀바귀의 생육)

  • Kim, Hye Min;Kang, Jeong Hwa;Jeong, Byoung Ryong;Hwang, Seung Jae
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.67-76
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    • 2016
  • This study was conducted to examine the optimal environmental condition for promoting the growth of sowthistle as affected by light quality and photoperiod in a closed-type plant production system. Seeds were sown in 240-cell plug trays and then germinated for 3 days at a 24-hour photoperiod in a closed-type plant production system with LED lights (R:B:W = 8:1:1). Seedlings were transplanted and grown under 3 types of LED (R:B:W = 8:1:1, R:W = 3:7, or R:B = 8:2) and 4 photoperiods (24/0, 16/8, 8/16, or 4/20 hours) with $230{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ light intensity at a density of $20cm{\times}20 cm$. The experimental design was a randomized complete block design. Plants were cultured for 40 days un der the condition of $21{\pm}2^{\circ}C$ and $70{\pm}10%$ relative humidity after transplanting. Plants were fed with a recycling nutrient solution (pH 7.0 and EC $2.0dS{\cdot}m^{-1}$) contained in a deep floating tank. Fresh weight and dry weight of shoot or root, leaf length, and leaf area were the greatest in the photoperiod of 24/0 (light/dark) with RW LED. The highest number of leaves occurred in the photoperiod of 16/8 (light/dark) with RB LED, while the incidence of tip burn was higher in the photoperiod of 24/0 (light/dark) compared to the other treatments. Chlorophyll value was the highest in the 16/8 (light/dark) photoperiod and there was no significant difference by light quality. Chlorophyll fluorescence was the lowest in the photoperiod of 24/0 (light/dark) compared with other treatments. Therefore, in terms of economic feasibility and productivity for Ixeris dentata Nakai cultivation in a closed-type plant production system, the results obtained suggest that plants grew the best when kept in a photoperiod of 16/8 (light/dark) and light quality of combined LED RW (3:7).

Signal Analysis of Optical Biosensor to Detect Peroxide Using Electrically Controlled Release System (전기적 방출 조절 시스템을 이용한 광 페록사이드센서의 개발)

  • Min, Jun-Hong;Lim, In-Hee;Kim, Hyo-Han;Lee, Sang-Beak;Choi, Jeong-Woo;Lee, Won-Hong
    • Journal of Sensor Science and Technology
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    • v.6 no.1
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    • pp.35-42
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    • 1997
  • The optical biosensor using the electrically controlled release of reactive reagent is developed for the detection of peroxide. Rapid degradation of polymer complex of PEOx and PMAA occurs as the applied current increases and thus released amount of HPA increases. The degradation velocity of polymer and the amount of HPA released are linearly proportional to the applied current. Peroxide is reacted with the released reagent by peroxidase and then the product, a fluorescent dimer DBDA, is formed. The monochromic light from light source (150W Xe arc ramp) excites the DBDA and the excited light is transmitted through an optical fiber to be detected by a photodiode array. The change of fluorescence intensity is related to the change of peroxide concentration. The peroxidase is entrapped in Ca-alginate get on the inner surface. The biosensor has the linear signal range of 0.025mM-10.mM peroxide. By applying the step function of peroxide, reproducibility of biosensor has been investigated. The mathematical model is constructed by the combination of enzyme kinetics with reactor flow model. Good agreement is obtained between the experimental result and model prediction in the sensor signal.

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Binding Site of Spermine at Poly$[d(A-T)_2]$ and Poly$[d(G-C)_2]$ (Poly$[d(A-T)_2]$, Poly$[d(G-C)_2]$와 스퍼민의 결합 형태에 관한 연구)

  • Yun, Byeong Hwa;Jeon, Sun Hee;Song, Young Dae;Cho, Tae Sub;Kim, Seog K.
    • Journal of the Korean Chemical Society
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    • v.42 no.5
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    • pp.506-511
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    • 1998
  • When the spermine, which is one of the polyamines containing cation in vivo, binds to DNA, it can increase the stability of DNA. At the same time, it can cause B-form to Z-form transformations of DNA. However, because we cannot determine the binding geometry of the spermine to DNA by using spectroscopic methods, nobody can show the accurate binding mechanism of a DNA-spermine complex. Thus, we used DAPI as a spectroscopic probe of spermine, which binding geometry was well known. At the result of base selective binding geometry of spermine to synthetic DNA, the concentration of spermine gets higher, it grows the hydrophobic environment of DAPI which bound the minor groove of adenine-thymine base pair. Simultaneously, spermine seems to bridge the backbones around the minor groove of $poly[d(A-T)_2]$. So that, the intensity of fluorescence spectrum of that shows sudden increasement. In guanine-cytocine base pair, $poly[d(G-C)_2]$, we can suppose that spermine bind to the major groove of that, shoving out the DAPI which is partially intercalated between the base pocket across the major groove of it. In both cases, spermine doesn't show the base selectivity against to DNA.

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CD11b as a Biomarker for Canine Systemic Inflammatory Response Syndrome and Sepsis (개 전신성염증반응증후군 및 패혈증의 진단적 표지자로서 CD11b의 활용)

  • Yu, Do-Hyeon;Noh, Dong-Ho;Song, Ru-Hui;Kim, Jun-Hwan;Lee, Da-Mi;Kim, Sue-Hee;Park, Chul;Park, Jin-Ho
    • Journal of Veterinary Clinics
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    • v.27 no.6
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    • pp.627-630
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    • 2010
  • The aim of this study is to investigate neutrophil activation markers among canine ICU (intensive care unit) control, systemic inflammatory response syndrome (SIRS) and sepsis. These markers include WBC (white blood cells), platelets counts, blood film examination (neutrophilic band to segmentation ratio and neutrophilic degenerative changes), and flow cytometric analysis (CD11b expression of neutrophils). As a result, the mean CD11b fluorescence intensity of neutrophils and the neutrophilic degenerative change scores were both significantly higher in sepsis group (P<0.05). In addition, mortality was also found to be correlated with the up-regulation of CD11b expression in circulating neutrophils. This study demonstrates that CD11b expression of neutrophils could be more a reliable biomarker to predict prognosis in ICU patients than traditional blood film examination according to this study.

Curcumin Attenuates Hydrogen Peroxide Induced Oxidative Stress on Semen Characteristics during In Vitro Storage of Boar Semen

  • Jang, Hyun-Yong;Kim, Young-Han;Cheong, Hee-Tae;Kim, Jong-Taek;Park, In-Chul;Park, Choon-Keun;Yang, Boo-Keun
    • Reproductive and Developmental Biology
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    • v.33 no.2
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    • pp.99-105
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    • 2009
  • Curcumin is a major active component of the food flovour tumeric. It has been used for the treatment of many diseases such as inflammatory and infectious diseases, cancer and other disease due to its antioxidant properties. Curcumin is a powerful scavenger of many free radicals such as superoxide anion, hydroxyl radical and nitric oxide. The objective of this study was to investigate the antioxidative effects of curcumin against hydrogen peroxide on semen quality during in vitro storage of boar semen. The sperm treated with different concentration of curcumin (1, 5 and 10 ${\mu}M$) in the presence or absence of hydrogen peroxide (250 ${\mu}M\;H_2O_2$) were incubated for 3, 6 and 9 hr at $37^{\circ}C$ and analyzed sperm characteristics such as motility, membrane integrity (MI), lipid peroxidation (LPO), reactive oxygen species (ROS) and DNA fragmentation (DF). The sperm motility and MI in $H_2O_2$ treated group ($47.8%{\pm}6.8$ and $24.8%{\pm}2.2$) were significantly decreased when compare to curcumin treated group ($79.8%{\pm}2.7$ and $34.6%{\pm}1.0$, respectively) irrespective of incubation periods(p<0.05). The LPO of spermatozoal plasma membrane was measured by thiobarbituric acid (TBA) reactions for malondialdehyde (MDA), MDA level in control ($11.6{\pm}0.6\;nmol/L{\times}10^6$) and curcumin groups ($10.7{\pm}0.3\;nmol/L{\times}10^6$) were lower than those of curcumin plus $H_2O_2$ ($17.1{\pm}0.8\;nmol/L{\times}10^6$) or $H_2O_2$ group ($22.5{\pm}1.9\;nmol/L{\times}10^6$) from 3 to 9 hr incubation periods. The DF by sperm chromatin dispersion (SCD) test and ROS production measured by 2',7'-dichlorofluorescein (DCF) fluorescence intensity were no significantly difference through all experimental groups (p>0.05). Correlation among evaluation methods for sperm quality, motility vs MI and DF vs ROS was positively correlated while motility vs DF and ROS vs LPO were negatively correlated in all treatment groups. These results demonstrate that curcumin can effectively improve the sperm quality during in vitro storage of boar semen through its hydrogen peroxide scavenging mechanism as an antioxidant.

Amplification of Porcine SRY Gene for Sex Determination

  • Choi, S.G.;Bae, M.S.;Lee, E.S.;Kim, S.O.;Kim, B.K.;Yang, J.H.;Jeon, C.E.;Kim, H.H.;Hwang, Y.J.;Lee, E.S.;Kim, D.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.8
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    • pp.1107-1112
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    • 2009
  • The separation of X and Y chromosome-bearing sperm is of use in many aspects of livestock maintenance. In this study, we sought to determine the difference in DNA content between X- and Y-bearing sperm, separate sperm into X- and Y-enriched pools, and assess the efficacy of sorting. Sperm collected from Duroc and miniature pigs were stained with 20.8 $\mu{M}$ Hoechst 33342 and analyzed using a high-speed cell sorter. Measurement of the fluorescence intensity of stained sperm nuclei revealed that the X-bearing sperm of Duroc and miniature pigs respectively contain 2.75% and 2.88% more DNA than Y-bearing sperm. In total, 50.18% of the sperm were assigned to the X-sorted sample and 49.82% was assigned to the Y-sorted sample for Duroc pigs. For miniature pigs, the Xsorted sample represented 50.19% of the population and the Y-sorted represented 49.81% of the population. Duplex PCR was used to evaluate accuracy of sorting. A fast and reliable method for porcine sexing was developed through amplification of the sex-determining region of the Y chromosome gene (SRY). Oligonucleotide primers were designed to amplify the conserved porcine SRY high motility group (HMG) box sequence motif. We found that the primer pair designed in this study was 1.46 times more specific than previously reported primers. Thus, this study shows that the present method can be applied in porcine breeding programs to facilitate manipulation of the sex ratio of offspring and to achieve precise sexing of porcine offspring by amplification of the HMG box of the SRY gene.

Determination of Terazocin in Human Plasma by Liquid Chromatography and Bioequivalence Study of Teratonin® Tablets (액체크로마토그래프법에 의한 사람 혈장 중 테라조신의 정량 및 테라토닌® 정의 생물학적 동등성)

  • Cho, Eun-Sook;Kang, Sung-Ha;Chun, In-Koo
    • Journal of Pharmaceutical Investigation
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    • v.32 no.2
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    • pp.119-125
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    • 2002
  • A rapid, selective and reproducible high-performance liquid chromatographic method has been developed for the determination of terazocin in human plasma. Terazocin plus the internal standard, prazocin hydrochloride, were extracted from alkalified plasma with tert-butylmethyl ether, back-extracted into 0.05% phosphoric acid. Fifty ${\mu}l-portions$ of extract were injected onto a octadecylsilane column and eluted with a mixture of acetonitrile, water and triethylamine (30 : 70 : 0.1 v/v, adjusted to pH 5.0 with dilute phosphoric acid) at a flow rate of 1.0 ml/min. The fluorescence intensity of column eluents was monitored at excitation wavelength of 250 nm and emission wavelength of 370 nm. No interference peaks were observed. The practical limit of quantitation was 5 ng/ml for terazocin. The average intraday and interday coefficients of variation were 4.15 and 3.54%, respectively. Also intraday and interday precisions over the range $5{\sim}60\;ng/ml$ were $0.49{\sim}2.92\;and\;0.38{\sim}5.12%$, respectively. The bioequivalence of two terazosin tablets, the $Hytrine^{\circledR}$ (Il Yang Pharmaceutical Co., Ltd.) and the $Teratonin^{\circledR}$ (Sam-A Pharmaceutical Co., Ltd.), was evaluated according to the guideline of Korea Food and Drug Administration (KFDA). Sixteen healthy male volunteers $(24.6{\pm}2.0\;years\;old)$ were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After one tablet containing 2 mg of terazosin was orally administered, blood was taken at predetermined time intervals and the concentration of terazosin in plasma was determined with a HPLC method using spectrofluorometric detector. AUC was calculated by the linear trapezoidal method. $C_{max}\;and\;T_{max}$ were compiled from the plasma drug concentration-time data. Analysis of variance (ANOVA) was utilized for the statistical analysis of the parameters. The results showed that the differences in $AUC_t,\;C_{max}\;and\;T_{max}$ between the two preparations were 0.21 %, 5.53% and 8.82%, respectively. The powers $(1-{\beta})\;for\;AUC_t,\;C_{max}\;and\;T_{max}$ were >99%, 97.49%, and 33.26%, respectively. Minimum detectable differences $({\Delta},\;%)\;at\;{\alpha}=0.1\;and\;1-{\beta}=0.8$ and the 90% confidence intervals were all less than ${\pm}20%$ except for $T_{max}.\;AUC_t\;and\;C_{max}$ met the criteria of KDFA for bioequivalence, indicating that $Teratonin^{circledR}$ tablets are bioequivalent to $Hytrine^{circledR}$ tablets.

Desalinated underground seawater of Jeju Island (Korea) improves lipid metabolism in mice fed diets containing high fat and increases antioxidant potential in t-BHP treated HepG2 cells

  • Noh, Jung-Ran;Gang, Gil-Tae;Kim, Yong-Hoon;Yang, Keum-Jin;Lee, Chul-Ho;Na, O-Su;Kim, Gi-Ju;Oh, Won-Keun;Lee, Young-Don
    • Nutrition Research and Practice
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    • v.4 no.1
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    • pp.3-10
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    • 2010
  • This study was performed to investigate the effect of desalinated underground seawater (named as 'magma seawater', MSW) of Jeju Island in Korea on lipid metabolism and antioxidant activity. MSW was collected from underground of Han-Dong in Jeju Island, and freely given to high fat diet (HFD)-fed C57BL/6 mice for 10 weeks. Although there were no significant differences in the body weight changes and plasma lipid levels, hepatic triglyceride levels were significantly lower in the MSW group than in the normal tap water (TW)-drunken control group. Furthermore, the activity of fatty acid synthase (FAS) was significantly decreased and carnitine palmitoyltransferase (CPT) activity was increased in MSW group compared to TW group. Similarly, real-time PCR analysis revealed that mRNA expressions of lipogenic genes were lowered in MSW groups compared to the control group. In a morphometric observation on the liver tissue, accumulation of fats was remarkably reduced in MSW group. Meanwhile, in vitro assay, tree radical scavenging activity measured by using diphenylpicrylhydrazyl (DPPH) was increased in MSW group. The 2'-7'-dichlorofluorescein diacetate (DCF-DA) staining followed with fluorescent microscopy showed a low intensity of fluorescence in MSW-treated HepG2 cells, compared to TW-treated HepG2 cells, which indicated that the production of reactive oxygen species by tert-butyl hydroperoxide (t-BHP) in HepG2 cells was decreased by MSW treatment. The antioxidant effect of MSW on t-BHP-induced oxidative stress in HepG2 cells was supported by the increased activities of intracellular antioxidant enzymes such as catalase and glutathione reductase. From these results, we speculate that MSW has an inhibitory effect on lipogenesis in liver and might play a protective role against cell damage by t-BHP-induced oxidative stress.