• 제목/요약/키워드: Fluorometry

검색결과 44건 처리시간 0.028초

P18 Nitric Oxide and Hypoxia Affect TCDD Induced EROD Activity

  • Kim, Yeo W.;Cha Y. Baek;Hong K. Min;Yhun. Y. Sheen
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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    • pp.138-138
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    • 1998
  • Effects of nitric oxide and hypoxia on ethoxyresorufin deethylase in Hepa I cells and MCF-7 human breast cancer cells were examined. TCDD treatment have resulted in the stimulation of ethoxyresorufin deethylase activity based on fluorometry in Hepa I in dose and time dependent manner. 0.1 nM TCDD showed maximal stimulation of ethoxyresorufin deethylase activity and 24 hour treatment also showed maximal stimulation of ethoxyresorufin deethylase activity. In MCF-7 human breast cancer cells, untreated cells showed high basal level of ethoxyresorufin deethylase activity. TCDD treatment to MCF-7 cells resulted minor stimulation of ethoxyresorufin deethylase activity compared to that in Hepa I cells. Nitric oxide and hypoxia inhibit TCDD effects on ethoxyresorufin deethylase activity in both cell lines. And also flavonoids, such as quercetin showed an inhibition of ethoxyresorufin deethylase activity that is stimulated with TCDD or 3-Methylcholanthrene. Estrogen and estrogen metabolite such as 16 a-estriol and 2-hydroxyestradiol also affects the ethoxyresorufin deethylase activity in MCF-7 cells.

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남서해역에서 양식되는 방사무늬김(Porphyra yezoensis Ueda)의 Diving-PAM에 의한 광합성 특성 (Photosynthetic Characteristics of Porphyra yezoensis Ueda Measured in situ by Diving Pulse-Amplitude Modulated (PAM) Fluorometry on the Southwestern Coast of the Korean Peninsula)

  • 김정배;이원찬;김형철;최희구;박정임;조윤식;박환희
    • 환경생물
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    • 제30권3호
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    • pp.210-218
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    • 2012
  • 일반적으로 많이 양식되는 방사무늬김(Porphyra yezoensis Ueda)에 대하여 방사무늬김 엽체의 형태적 특성, 탄소 및 질소 성분의 농도, 안정동위원소 비값, 광합성 효율을 방사무늬김 양식이 이루어 지고 있는 남서해역에서 조사를 하였다. 방사무늬김의 형태적 특성에 대해서 살펴보면 평균 엽장은 11.6~16.3 (13.8) cm, 평균 엽폭은 4.6~6.3 (5.4) cm이었고, 단위면적당 방사무늬김 엽체의 평균 엽중량은 $1.1{\sim}2.6(1.86)g\;DW\;m^{-2}$이었다. 단위면적당 Chl a 농도는 2.18~17.77 (평균 9.65) mg DW Chl a $m^{-2}$이었다. 방사무늬김의 탄소 농도는 $201{\sim}317(240)mg\;DW\;g^{-1}$이었고, 질소 농도는 $39.8{\sim}50.0(43.5)mg\;DW\;g^{-1}$이었으며, C/N비는 5.0~6.7 (5.5)이었다. 방사무늬김의 방사성 안정동위원소비 중에서 탄소 안정동위원소 비는 ${\delta}^{13}C$=-25.6‰ 에서 ${\delta}^{13}C$=-24.0‰ (평균 -24.7‰)의 값을 보였고, 질소 안정동위원소 비는 ${\delta}^{15}N$=1.3‰ 에서 ${\delta}^{15}N$=4.1‰ (평균 2.1‰)의 값을 보였다. PAM에 의한 해조류의 광합성 특성은 광합성 활동의 지시자로서 사용될 수 있다. 우리는 Diving-PAM을 이용하여 각 정점 해조류인 방사무늬김의 광합성율을 분석하였다. 최대양자수율은 0.46~0.55 (평균 0.52)로서 최대 양자수율의 변동은 정점간 큰 차이는 없었다. 최대상대전자전달률은 4.71~5.84 (평균 5.33) ${\mu}mol\;electrons\;m^{-2}\;s^{-1}$ 로서 최대양자수율과 비슷한 분포를 보였다. 기울기 (${\alpha}$)는 0.027~0.045 (평균 0.036)을 보였고, 전자전달을 위한 포화광은 지역에 따라서 일부 차이를 보였으나 $139{\sim}180(156){\mu}mol\;photons\;m^{-2}\;s^{-1}$이었다. 남서해역 방사무늬김 엽체의 탄소 및 질소 농도와 광합성 효율은 지역에 따른 큰 차이는 보이지 않았다. 광합성 특성은 낮은 최대양자수율과 최대 상대전자전달률로 인한 낮은 광합성 효율이 나타났다.

Tetracycline Inducible Retrovirus Vector System에 의한 GFP 유전자의 발현 조절 (Regulation of GFP Expression Using the Tetracycline Inducible Retroviral Vector System)

  • 구본철;권모선;김태완
    • Reproductive and Developmental Biology
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    • 제29권1호
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    • pp.57-62
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    • 2005
  • 본 연구에서는 retrovirus를 이용한 유전자 전이에 있어서 대두되는 큰 문제점의 하나인 외래 유전자의 지속적인 발현으로 인한 개체의 생리적인 손상을 최소화하기 위하여 tetracycline계 물질의 공급 여부에 따라서 발현을 유도적으로 조절할 수 있는 one vector 형태의 Tet-On system을 구축하고자 하였다. 또한 WPRE 서열을 이 vector 상에 도입하여 유도적 조건에서 외래 유전자의 발현이 보다 강하게 일어날 수 있는 효율적인 retrovirus vector system을 확립하고자 하였다. 구축한 각각의 vector system에서 fluorometry와 western blotting을 이용하여 GFP 유전자의 발현 정도를 비교 측정한 결과, RevTRE-EGFP-WPRE-RSVp-rtTA2SM2 virus를 이용하여 유전자를 전이시킨 표적세포에서 GFP의 절대적인 발현량이 가장 큰 것으로 나타났고, 유전자 발현의 turn on/off에 의한 유도율은 RevTRE-EGFP-RSVp-rtTA2SM2-WPRE virus의 경우에서 8∼21배로 가장 높은 것으로 나타났다. 이상의 결과에서 외래 유전자의 발현을 효율적으로 조절할 수 있는 vector system은 WPRE가 rtTA2SM2 서열의 3에 위치한 형태로, 이 system을 이용하여 생산한 고감염가의 virus는 유전자 치료나 형질전환 동물의 생산에 있어서 요구되는 외래 유전자의 발현을 효율적으로 조절할 수 있는 수단이 될 것이다.

수용성계의 Linoleic Acid와 LDL에 대한 한국산 홍삼의 산화방지효과 (Antioxidant Effect of Korean Red Ginseng Extract on Aqueous Linoleic Acid and LDL)

  • 이종원;이성계;도재호;성현순;이형옥
    • Applied Biological Chemistry
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    • 제40권4호
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    • pp.283-288
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    • 1997
  • 1% linoleic acid를 함유하는 수용성 완충용액과 LDL(low-density lipoprotein, 1 mg protein/ml)을 함유하는 수용성 완충용액 각각에 $H_2O_2$$FeCl_2$를 가하여 유발된 산화과정 중 한국산 홍삼 extract(red ginseng extract; RGE)에 대한 산화방지활성을 비교물질로 ${\alpha}-tocopherol$을 사용하여, HPLC를 이용한 MDA(malondialdehyde) 정량법과 fluorometry를 이용하여 측정 비교하였다. LDL(0.25 mg protein/ml)에서 생성된 conjugated diene도 spectrometry로 측정하였다. Linoleic acid를 기질로 사용한 경우, MDA 생성량으로 비교한 산화방지효과에 있어서 1000 ppm RGE의 첨가로 71.8%의 우수한 산화저해율이 나타났으며, 이때 비교물질로 사용한 100 ppm ${\alpha}-tocopherol$의 경우에는 76.1%이었다. LDL(1 mg/ml)을 기질로 사용하였을 경우에는 RGE 200 ppm 첨가시 가장 우수한 항산화효과가 나타났으며, 이때의 산화저해율은 25.2%이었고, 100 ppm ${\alpha}-tocopherol$의 경우는 21.2%이었다. 또한 LDL(0.25 mg protein/ml)을 기질로 사용하여 생성된 c-diene을 측정한 경우에도 50 ppm RGE의 첨가로 44.2% 저해효과가 입증되었다.

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Contents of Phytic Acid and Minerals of Rice Cultivars from Korea

  • Lee, Heok-Hwa;Rhee, Hae-ik;Lee, Sang-Young;Kim, Chon-Ho;Park, Yong-Soon
    • Preventive Nutrition and Food Science
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    • 제2권4호
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    • pp.301-303
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    • 1997
  • To elucidate the contents of phytic acid and minerals in rice produced in Korea, samples of 68 verieties of brown rice and 9 varieties of polished rice were analyzed for phytic acid by colorimetric method, and Ca, Fe, Mg and Zn contents by the atomic absorption spectrophytometer. Selenium was measured by fluorometry. Averages of phytic acid content of the 68 brown rice were 12.6 g/kg, whereas that of the 8 polished rice were 1.83 g/kg on dry matter. Averages of Ca, Fe, Mg and Zn contents on the 68 brown rice were 111, 7.4, 1,068 and 19.1 mg/kg, and those of the polished rice were 45.6, 2.1, 250 and 14 mg/kg, respectively. A averge of selenium content of the brown rice was 38.3 $\mu\textrm{g}$/kg.

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STUDY CYTOCHROME P450IA1 GENE EXPRESSION BY RTPCR.

  • Lee, Soo-Young;Yhun Y. Sheen
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1995년도 춘계학술대회
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    • pp.128-128
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    • 1995
  • To investigate the mechanism of the regulation of cytochrome P450IA1 gene expression, ethoxyresorufin deethylase(EROD) and benzo(a)pyrene hydroxylase in B6 mouse liver, in isolated perfused rat liver system. and in B6 mouse hepatocyte Hepa-I cells were examined. In C57BL/6N mouse, 3-methylcholan- throne( 3MC ) treatment have resulted in the stimulation of EROD activity based on fluorometry by 2.79 fold comparirng with that of control. Measurement of mRNA of cytochrome P450 was carried out by either nothern blot or dot blot analysis. Findings are similar to that of studies with enzymes. Furhtermore, when RTPCR method was applied to detect mRNA in Hepa I cell and liver tissues the results were more clear. Cytochrome P450IA1 upstream DNA containing CAT construct was transfected into Hepa-1 cells. After transfection of CAT construct, 3MC and flavonoids, such as, chrysin, hesperetin, kaempferol, morin, myricetin and aminoyrine were treated. 48 Hours after treatments, cells were harvested and assayed for CAT mRNA by RTPCR. 3MC treatment to hepa I cells transfected with trout P450IA1-CAT construct increased CAT mRNA by 2.81 fold when it was compared with that of control. This increase CAT mRNA was decreased by concomitantly treated flavonoids and aminopyrine. The level of CAT protein was 29.2-58.0% of 3MC stimulated CAT protein. Results of this study suggested that RTPCR seems to be a very good method to study regulation of gene expression in liver tissue or Hepa cells.

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Form, Function and Longevity in Fucoid Thalli: Chlorophyll a Fluorescence Differentiation of Ascophyllum nodosum, Fucus vesiculosus and F. distichus (Phaeophyceae)

  • Kim, Kwang-Young;Garbary, David j
    • ALGAE
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    • 제24권2호
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    • pp.93-104
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    • 2009
  • Imaging-PAM fluorometry was used to assess the chlorophyll a fluorescence parameter ${\Phi}_{PSII}$ (effective quantum yield) in Frcus vesiculosus. F. disttchus. ssp. distichus and AscophyIIum nodosum. The objective was to show variadon in fluorescence yield associated with age and frond organ, and to illustrate the spatial scales at which photosynthetic parameters vary on fucoid thalli. In addition, our species represented taxa in different but related genera, species with different ecoloeies (rock pool and non rock pool species), morphologies (with and without air bladders) and longevities (several to 20 or more years). A further objective was to determine the extent to which photosynthetic parameters reflected these differences- Effective quantum yield declined substantially with age in F. vesiculosus and F. distichus ssp. distichus, whereas ${\Phi}_{PSII}$ in A. nodosum was maximal after three years. In A. nodosum ${\Phi}_{PSII}$ was still high in branch segments at least seven years old. Older branches of A. nodosum showed relatively higher and more homogeneous photosynthetic capacity relative to Fucus species. Surfaces of air bladders in A. nodosum and F. vesicu- losus had ${\Phi}_{PSII}$ that was not significantly different from the highest rates, achieved in these species. The heterogene- ity of photosynthetic efficiency is consistent with morphological and developmental differences among the species and their ecology. in particular the longevity of A. nodosum fronds.

복분자 열매의 총 페놀성분의 정량 및 항산화 활성 (Determination of Total Phenolic Compounds from the Fruit of Rubus coreanum and Antioxidative Activity)

  • 이종원;도재호
    • 한국식품영양과학회지
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    • 제29권5호
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    • pp.943-947
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    • 2000
  • The purpose of this study was to investigate the extraction method of phenolic compounds from Rubus coreanum and antioxidative activity. antioxidative activities of Rubus coreanum were tested with ability of donating hydrogen to DPPH, and HPLC, fluorometry which measure the amount of MDA after reacting linoleic acid with $H_2O$$_2$, and LDL with $H_2O$$_2$ and FeCl$_2$. The most suitable extraction conditions of the phenolic compounds from Rubus coreanum was 3 times with 60% ethanol, and the yield of extract containing 35% moisture was 15.28%. In extraction efficacy of phenolic compounds, 60% ethanol was superior to water as extraction solvent, and extraction efficacy with 60% ethanol did not differ from disolving by water after evaporation of 60% ethanol extract. 60% ethanol extract of Rubus coreanum had an ability of hydrogen donating to DPPH, MDA determination showed the antioxidative effect with inhibition ratio of 77.91% on linoleic acid oxidation by addition of Rubus coreanum extract with the concentration of 1.500 ppm. and about 65.74% of LDL oxidation was inhibited by addition of 1,000 ppm.

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Fluorescence Spectroscopy Studies on Micellization of Poloxamer 407 Solution

  • Lee, Ka-Young;Shin, Sang-Chul;Oh, In-Joon
    • Archives of Pharmacal Research
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    • 제26권8호
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    • pp.653-658
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    • 2003
  • It has been reported that at low temperature region, poloxamers existed as a monomer. Upon warming, an equilibrium between unimers and micelles was established, and finally micelle aggregates were formed at higher temperature. In this study, the fluorescence spectroscopy was used to study the micelle formation of the poloxamer 407 in aqueous solution. The excitation and emission spectra of pyrene, a fluorescence probe, were measured as a function of the concentration of poloxamer 407 and temperature. A blue shift in the emission spectrum and a red shift in the excitation spectrum were observed as pyrene transferred from an aqueous to a hydrophobic micellar environment. From the $I_1/I_3 and I_{339}/I_{333}$ results, critical micelle concentration (cmc) and critical micelle temperature (cmt) were determined. Also, from the fluorescence spectra of the probe molecules such as 8-anilino-1-naphthalene sulfonic acid and 1-pyrenecarboxaldehyde, the blue shift of the $\lambda_{max}$ was observed. These results suggest a decrease in the polarity of the microenvironment around probe because of micelle formation. The poloxamer 407 above cmc strongly complexed with hydrophobic fluorescent probes and the binding constant of complex increased with increasing the hydrophobicity of the probe.

Serum Kynurenic Acid: Possible Association with Invasiveness of Non-small Cell Lung Cancer

  • Sagan, Dariusz;Kocki, Tomasz;Kocki, Janusz;Szumilo, Justyna
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권9호
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    • pp.4241-4244
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    • 2012
  • The lung adenocarcinoma is considered more aggressive than other types of non-small cell lung cancer. As metabolites of tryptophan degradation along the kynurenine pathway, including kynurenic acid, have been shown to induce immunosuppression and facilitate escape of tumor cells from immune surveillance, a hypothesis was set up that differences in biological behavior between types of lung cancer may be associated with altered activity of the kynurenine metabolic pathway. The aim of the study was to determine kynurenic acid levels in the serum of patients with bronchial adenocarcinoma for comparison with other types of non-small cell lung cancer. A total of 227 patients with non-small cell lung cancer were enrolled in the study, including 71 with adenocarcinoma and 96 with squamous cell carcinoma. Serum kynurenic acid concentration was determined with use of high performance liquid chromatography and fluorometry. The level of kynurenic acid in the serum of patients with adenocarcinoma was significantly higher than in those with squamous cell lung cancer ($107.1{\pm}62.8$ pmol/ml; 95%CI: 92.4 to 132.3 pmol/ml versus $82.1{\pm}47.6$ pmol/ml; 95%CI: 78.5 to 91.2 pmol/ml, respectively; p = 0.027). Differences between other histological types of lung cancer were insignificant. We conclude that increased activity of kynurenine metabolic pathway manifested by elevated serum kynurenic acid level may be one of the factors associated with clinically distinct biological behavior of adenocarcinoma, in particular high invasiveness and rapid progression.