• Title/Summary/Keyword: Fluorescence life times

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Amphiphilic graft copolymers: Effect of graft chain length and content on colloid gel

  • Nitta, Kyohei;Kimoto, Atsushi;Watanabe, Junji;Ikeda, Yoshiyuki
    • Biomaterials and Biomechanics in Bioengineering
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    • v.2 no.2
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    • pp.97-109
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    • 2015
  • A series of amphiphilic graft copolymers were synthesized by varying the number of graft chains and graft chain lengths. The polarity of the hydrophobic graft chain on the copolymers was varied their solution properties. The glass transition temperature of the copolymers was in the low-temperature region, because of the amorphous nature of poly (trimethylene carbonate) (PTMC). The surface morphology of the lyophilized colloid gel had a bundle structure, which was derived from the combination of poly(N-hydroxyethylacrylamide)( poly(HEAA)) and PTMC. The solution properties were evaluated using dynamic light scattering and fluorescence measurements. The particle size of the graft copolymers was about 30-300 nm. The graft copolymers with a higher number of repeating units attributed to the TMC (trimethylene carbonate) component and with a lower macromonomer ratio showed high thermal stability. The critical association concentration was estimated to be between $2.2{\times}10^{-3}$ and $8.9{\times}10^{-2}mg/mL$, using the pyrene-based fluorescence probe technique. These results showed that the hydrophobic chain of the graft copolymer having a long PTMC segment had a low polarity, dependent on the number of repeating units of TMC and the macromonomer composition ratio. These results demonstrated that a higher number of repeating units of TMC, with a lower macromonomer composition, was preferable for molecular encapsulation.

Comparison of Conditions for Cell Death-Inducing Agents Using a High Throughput-Compatible Nuclear Staining Assay (핵 염색을 이용한 세포사멸 유도물질 스크리닝의 조건 비교)

  • Lee, Sang-Han
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1312-1315
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    • 2008
  • High throughput-drug screening plays a pivotal role for early stage of drug discovery process. In the course of assay development for screening of cell death-inducing agents, a protocol that is simple, time-saving, and high throughput-compatible was designed which was confirmed that the protocol can be worked by a HTS-compatible machine. In 96-well format, PC-3 cancer cells (1${\times}10^{4}$ cells/ml) were cultured for 24 hr. After 24 h-incubation with various medicinal plants extracts, the cells were then stained with DAPI for 30 min. The fluorescence intensity of the stained cells was measured semi-automatically with a multilabel counter. To test whether the present assay system effectively works, we screened about 850 medicinal plant extracts, and selected 1 positive crude extracts that contained cell death-inducing activity. These results suggest that the protocol is highly amenable to HTS implementation for a cell death-inducing agent(s).

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.

Analysis of the Behavior of Fluorescent Whitening Agents in Recycling Process of White Ledger (형광증백제가 함유된 백상고지의 재활용에 따른 형광증백제의 거동 분석)

  • Lee, Ji Young;Kim, Chul Hwan;Park, Jong-Hye;Kim, Eun-Hea;Sung, Yong Joo;Heo, Young-Jun;Kim, Young-Hoon;Kim, Yeon-Oh
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.47 no.1
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    • pp.52-58
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    • 2015
  • White ledger usually includes white office paper, computer paper, and copy machine paper. Because these grades need high optical properties, fluorescent whitening agents (FWAs) are widely used in the papermaking process. FWAs are the most powerful and effective chemical used to obtain high CIE whiteness and ISO brightness in papers. The rising demand for white or ultra-white papers has increased the use of FWAs. However, FWAs used in white ledger can restrict its use, even though white ledger is widely used as a raw material in paperboard mills. Therefore, it is necessary to develop methods to control FWAs from white ledger to increase its use in paperboard mills. In this study, the behaviors of disulpho fluorescent whitening agent (D-FWA), tetrasulpo fluorescent whitening agent (T-FWA), and hexasulpho fluorescent whitening agent (H-FWA) during the recycling process were identified as a first step to remove FWAs from white ledger. We prepared four types of papers (dyed with D-FWA, T-FWA, and H-FWA), disintegrated these papers, and made handsheets. This recycling process was carried out three times in a laboratory. After each round of recycling, the hand-sheets' CIE whiteness and fluorescence index were measured, and the distribution of FWAs in the Z-direction was observed using CLSM images. FWA reductions in the model papers were calculated using fluorescence indices as a function of the number of recycling. FWAs in handsheets containing T-FWA and H-FWA decreased linearly as a function of the number of recycling, but D-FWA did not show a significant reduction in the fluorescence index after recycling. T-FWA and H-FWA showed similar distributions of D-FWA after recycling. Therefore, as much T-FWA and H-FWA as possible must be detached in the early processes of papermaking at paperboard mills.

Determination of Abamectin Residue in Paprika by High-Performance Liquid Chromatography

  • Xie, Wen-Ming;Ko, Kwang-Yong;Kim, Sung-Hun;Chang, Hee-Ra;Lee, Kyu-Seung
    • Korean Journal of Environmental Agriculture
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    • v.25 no.4
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    • pp.359-364
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    • 2006
  • Reversed-phase high-performance liquid chromatography (HPLC) techniques were developed to quantify abamectin (ABM) in paprika (Capsicum annum). Separation was achieved on a $C_{18}$ ODS column with a mobile phase of acetonitrile/water (96/4, v/v) mixture in an isocratic elution at the flow tate of 1.2 mL/min for avermectins (AVMs). The retention times were 8.0 and 9.7mins for AVM $B_{lb}$ and AVM $B_{1a}$, respectively. Residual AVMs (sum of AVM $B_{1a}$, AVM $B_{1b}$ and 8,9-Z-AVM $B_{1a}$) in the vegetable were extracted with acetonitrile, and the silica solid-phase extraction cartridges were used to purify the extract. AVMs were derivatized using trifluoroacetic acid and 1-methylimidazole, and the derivatives were determined with a fluorescence detector (excitation at 365 nm and emission at 470 nm). High and consistent recoveries, ranging from 93% to 115%, were obtained for AVM $B_{1a}$ and 8, 9-Z-AVM $B_{1a}$ at fortified levels of $20{\mu}g/kg\;and\;200{\mu}g/kg$ for paprika. The limit of quantitation (LOQ) was $2{\mu}g/kg$. The residual levels of AVMs in paprika in a field experiment from one day to seven days after the last application decreased from 18.40 to $7.59{\mu}g/kg$. The half-life $(T_{1/2})$ of AVMs in paprika was 1.47 days.

An Advanced Kinetic Method for HO2·/O2-· Determination by Using Terephthalate in the Aqueous Solution

  • Kwon, Bum Gun;Kim, Jong-Oh;Kwon, Joong-Keun
    • Environmental Engineering Research
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    • v.17 no.4
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    • pp.205-210
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    • 2012
  • Hydroperoxyl radical/superoxide anion radical ($HO_2{\cdot}/O_2^-{\cdot}$, $pK_a$=4.8) as an intermediate is of considerable importance in oxidation processes. Hence, the method of detecting $HO_2{\cdot}/O_2^-{\cdot}$ with high sensitivity is necessary to be developed. To achieve this objective, this study newly employed terephthalate (TA) as a probe for the measurement of $HO_2{\cdot}/O_2^-{\cdot}$ in the kinetic method presented in our previous study. This method was based on the hydroxylation of TA to produce mainly hydroxyterephthalic acid or hydroxyterephthalate (OHTA), which was analyzed by fluorescence detection (${\lambda}_{ex}$=315nm, ${\lambda}_{ex}$=425nm). The life-time of $HO_2{\cdot}/O_2^-{\cdot}$ and its concentration formed from the photolysis technique of $H_2O_2$ were reported in this study. At range of pH 2-10, the life-time of $HO_2{\cdot}/O_2^-{\cdot}$ was 51-422 sec. In particular, an increase in the life-time with pH was observed. The sensitivities of the kinetic method by using TA were always higher with 1.7-2.5 times at pH 8.0 than those by using benzoic acid. From these results, this study can contribute to understanding the basic functions of $HO_2{\cdot}/O_2^-{\cdot}$ in oxidation processes.

Determination of Hydroperoxyl/superoxide Anion Radical (HO2·/O2·-) Concentration in the Decomposition of Ozone Using a Kinetic Method

  • Kwon, Bum-Gun;Lee, Jai H.
    • Bulletin of the Korean Chemical Society
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    • v.27 no.11
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    • pp.1785-1790
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    • 2006
  • A novel kinetic method for determination of $HO_2{^{\cdot}}/O_2{^{{\cdot}-}}$ in ozone decomposition in water is described. In this study, potential interferences of $O_3$ and the hydroxyl radicals, $^{\cdot}OH_{(O3)}$, are suppressed by $HSO_3{^-}/SO_3{^{2-}}$. $HO_2{^{\cdot}}/O_2{^{{\cdot}-}}$ formed in ozone decomposition reduces $Fe^{3+}$-EDTA into $Fe^{2+}$-EDTA and subsequently the well-known Fenton-like (FL) reaction of $H_2O_2$ and $Fe^{2+}$-EDTA produces the hydroxyl radicals, $^{\cdot}OH_{(FL)}$. Benzoic acid (BA) scavenges $^{\cdot}OH_{(FL)}$ to produce OHBA, which are analyzed by fluorescence detection (${\lambda}_{ex}=320nm$ and ${\lambda}_{ex}=400nm$). The concentration of $HO_2{^{\cdot}}/O_2{^{{\cdot}-}}$ in ozone decomposition has been determined by the novel kinetic method using the experimentally determined half-life ($t_{1/2}$). The steady-state concentration of $HO_2{^{\cdot}}/O_2{^{{\cdot}-}}$ is proportional to the $O_3$ concentration at a given pH. However, the steady-state concentration of $HO_2{^{\cdot}}/O_2{^{{\cdot}-}}$ in ozone decomposition is inversely proportional to pH values. This pH dependence is due to significant loss of $O_2{^{{\cdot}-}}$ by $O_3$ at higher pH conditions. The steady-state concentrations of $HO_2{^{\cdot}}/O_2{^{{\cdot}-}}$ are in the range of $2.49({\pm}0.10){\times}10^{-9}M(pH=4.17){\sim}3.01({\pm}0.07){\times}10^{-10}M(pH=7.59)$ at $[O_3]_o=60{\mu}M$.

Optical Spectroscopy of$Eu^{3+}$in Borosilicate Glasses (붕규산유리 매질 내에서의$Eu^{3+}$이온의 형광특성)

  • Kim, Jong Goo;Lee, Jong Gyu;Kim, Jung Suk;Kim, Seung Soo;Chun, Kwan Sik
    • Journal of the Korean Chemical Society
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    • v.41 no.5
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    • pp.251-255
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    • 1997
  • Fluorescence spectra and life time of $Eu^{3+}$ ion in borosilicate glass medium are measured. Electronic transitions of $Eu^{3+}$ ion in borosilicate glass medium are found to come from $5D0{\rightarrow}7FJ$(J=0, 1, 2, 3, 4) transitions of SL coupling system in $f^b$ electrons configuration. From the number of Stark sublevels in spetra, crystal field for $Eu^{3+}$ ion is also found to have the symmetric character of low symmetry order, $n{\leq}2$. The fraction and the number of components of life times were varied depending on the composition of $Eu^{3+}$ in borosilicate glasses, from which the binding condition between the $Eu^{3+}$ ion and anionic oxygen of borosilicate glass can be deduced.

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Studies on the Nuclei Adduction and Expression of c-myc Gene by Benzo(a)pyrene and Doxorubicin in Human NC-37 Cells (사람 NC-37 세포에서 Benzo(a)pyrene과 Doxorubicin에 의한 Nuclei내전과 c-myc 유전자의 발현에 대한 연구)

  • 김호찬;정인철;조무연
    • Journal of Life Science
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    • v.8 no.4
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    • pp.400-409
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    • 1998
  • Formation of adduct was studied in benzo(a)pyrene(BP)- and doxorubicin(Dx)-treated human NC-37 cells and isolated nuclei. Major adducts formed were determined by fluorescence absorption spectrophotometery and DNA-lin-ked protein assay. When isolated nuclei were exposed to carcinogens BP and DMBA, and anticancer drugs m-AMSA, ellipticine and Dx, varying degrees of adduct formation occured between DNA-protein complex and these drugs. When the mixture was centrifuged 1.7 M sucrose solution, binding BP and DMBA appeared to be similar between the sediment and the supernatant. When the sediment was centrifuged again with 0.35% polymin-P, the amount of BP bound was 2-fold greater in the protein(1077$\pm$55cpm) than in DNA fraction (470$\pm$20cpm), whereas that of DMBA was 1.6-fold greater in the DNA than in protein fraction. In the case of m-AMSA, ellipticine and Dx, the amount of binding was slightly greater in supernatant than in sediment in centrifugation with 1.7 M sucrose, and more than 3 times greater in the DNA- than in protein- fraction in centrifugation with 0.35% polymin P. DNA fractions which associated with a subset of nonhistone chromosomal protein were isolated from NC-37 cells exposed to $^{3}$H-BP and $^{14}$C-Dx. They were separated into two distince components DNA-S and DNA-P by centrifugation with 2M Nacl chromatin extraction. The results indicated that the amount of $^{3}$H-BP bound was 6.0-fold greater in DNA-P as compared with DNA-S, while that of $^{14}$C-Dx binding appreaed to be 6.2-fold greater in DNA-S than in DNA-P fraction. When $^{3}$H-BP binding wasdetermined in the presence of cold Dx, the amount of binding was reduced only in the DNA-P fraction, indicating that the interaction between DNA and protein is decreased. Gene expression by these drugs, BP treated cells were increased to compare with nomal cells but reduced by treatment with BP-Dx. These results suggest that the protein moiety which tightly bound to DNA-P fraction may play an important role in the regulation of gene expression.

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