• Title/Summary/Keyword: 침전제

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The Change of Cell-cycle Related Proteins and Tumor Suppressive Effect in Non-small Cell Lung Cancer Cell Line after Transfection of p16(MTS1) Gene (폐암세포에 p16 (MTS1) 유전자 주입후 암생성능의 변화 및 세포주기관련 단백질의 변동에 관한 연구)

  • Kim, Young-Whan;Kim, Jae-Yeol;Yoo, Chul-Gyu;Han, Sung-Koo;Shim, Young-Soo;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.796-805
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    • 1997
  • Background : It is clear that deregulation of cell cycle progression is a hallmark of neoplastic transformation and genes involved in the $G_1$/S transition of the cell cycle are especially frequent targets for mutations in human cancers, including lung cancer. p16 gene product, one of the G1 cell-cycle related proteins, that is recently identified plays an important role in the negative regulation of the the kinase activity of the cyclin dependent kinase (cdk) enzymes. Therefore p16 gene is known to be an important tumor suppressor gene and is also called MTS1 (multiple tumor suppressor 1). No more oncogenes have been reported to be frequently related to multiple different malignancies than the alterations of p16 gene. Especially when it comes to non-small cell lung cancer, there was no expression of p16 in more than 70% of cell lines examined. And also it is speculated that p16 gene could exert a key role in the development of non-small cell lung cancer. This study was designed to evaluate whether p16 gene could be used as a candidate for gene therapy of non-small cell lung cancer. Methods : After the extraction of total RNA from normal fibroblast cell line and subsequent reverse transcriptase reaction and polymerase chain reaction, the amplified p16 cDNA was subcloned into eukaryotic expression plasmid vector, pRC-CMV. The constructed pRC-CMV-p16 was transfected into the NCI-H441 NSCLC cell line using lipofectin. The changes of G1 cell-cycle related proteins were investigated with Western blot analysis and immunoprecipitation after extraction of proteins from cell lysates and tumor suppressive effect was observed by clonogenic assay. Results : (1) p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 showed the formation of p16 : cdk 4 complex and decreased phosphorylated Rb protein, while control cell line did not. (2) Clonogenic assay demonstrated that the number of colony formation was markedly decreased in p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 than the control cell line. Conclusion : It is confirmed that the expression of p16 protein in p16 absent NSCLC cell line with the gene transfection leads to p16 : cdk4 complex formation, subsequent decrease of phosphorylated pRb protein and ultimately tumor suppressive effects. And also it provides the foundation for the application of p16 gene as a important candidate for the gene therapy of NSCLC.

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Purification of Complement System-Activating Polysaccharide from Hot Water Extract of Young Stems of Cinnamomum cassia Blume (계지(桂枝) 열수추출물로부터 보체계 활성화 다당의 정제)

  • Kweon, Mee-Hyang;An, Hyun-Jung;Shin, Kwang-Soon;Na, Gyeong-Su;Sung, Ha-Chin;Yang, Han-Chul
    • Korean Journal of Food Science and Technology
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    • v.29 no.1
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    • pp.1-8
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    • 1997
  • A complement system-activating (anti-complementary) polysaccharide was purified from the hot water extract of young stems of Cinnamomum cassia Blume. Crude polysaccharide fraction (CC-1) was prepared from the hot water extract of the young stems followed by methanol-reflux, precipitation with ethanol, dialysis, and lyophilization. The anti-complementary activity of CC-1 was decreased greatly by periodate oxidation, but was not changed by pronase digestion. These suggest that carbohydrate moiety may be related to the activation of complement system. According to its ionic strength CC-1 was fractionated first using cetavlon to give 4 fractions, CC-2, 3, 4 and 5. Among them CC-2 fraction was found to retain the highest activity and yield. CC-2 was separated to an unabsorbed neutral sugar portion (CC-2-I) and seven absorbed acidic sugar fractions $(CC-2-II{\rightarrow}CC-2-VIII)$ on DEAE-Toyopearl 650C (Cl-). CC-2-III showing higher anti-complementary activity and yield than those of other fractions, was further purified on the gel permeation of Sephadex G-100 and Sepharose CL-6B to CC-2-IIIa-3. CC-2-IIIa-3 was determined to have a homogeneity hy GPC (Sepharose CL-6B) and HPLC. Gel chromatography using standard dextrans gave a value of $2.4{\times}10^5$ for the molecular weight. The purified polysaccharide, CC-2-IIIa-3 consisted of arabinose, xylose, glucose, galactose, galacturonic acid and glucuronic acid in a molar ratio of 5.56 : 3.77 : 1.87 : 1.00 : 5.12 : 3.13 and contained no nitrogen.

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Toxicity Assessment of Silver Ions Compared to Silver Nanoparticles in Aqueous Solutions and Soils Using Microtox Bioassay (Microtox 생물검정법을 이용한 은 이온과 은 나노입자의 수용액과 토양에서의 독성 비교 평가)

  • Wie, Min-A;Oh, Se-Jin;Kim, Sung-Chul;Kim, Rog-Young;Lee, Sang-Phil;Kim, Won-Il;Yang, Jae E.
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.6
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    • pp.1114-1119
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    • 2012
  • This study was conducted to assess the microbial toxicity of ionic silver solution ($Ag^+N$) and silver nanoparticle suspension ($Ag^0NP$) based on the Microtox bioassay. In this test, the light inhibition of luminescent bacteria was measured after 15 and 30 min exposure to aqueous solutions and soils spiked with a dilution series of $Ag^+N$ and $Ag^0NP$. The resulting dose-response curves were used to derive effective concentration (EC25, $EC_{50}$, EC75) and effective dose ($ED_{25}$, $ED_{50}$, $ED_{75}$) that caused a 25, 50 and 75% inhibition of luminescence. In aqueous solutions, $EC_{50}$ value of $Ag^+N$ after 15 min exposure was determined to be < $2mg\;L^{-1}$ and remarkably lower than $EC_{50}$ value of $Ag^0NP$ with $251mg\;L^{-1}$. This revealed that $Ag^+N$ was more toxic to luminescent bacteria than $Ag^0NP$. In soil extracts, however, $ED_{50}$ value of $Ag^+N$ with 196 mg kg-1 was higher than $ED_{50}$ value of $Ag^0NP$ with $104mg\;kg^{-1}$, indicating less toxicity of $Ag^+N$ in soils. The reduced toxicity of $Ag^+N$ in soils can be attributed to a partial adsorption of ionic $Ag^+$ on soil colloids and humic acid as well as a partial formation of insoluble AgCl with NaCl of Microtox diluent. This resulted in lower concentration of active Ag in soil extracts obtained after 1 hour shaking with $Ag^+N$ than that spiked with $Ag^0NP$. With longer exposure time, EC and ED values of both $Ag^+N$ and $Ag^0NP$ decreased, so their toxicity increased. The toxic characteristics of silver nanomaterials were different depending on existing form of Ag ($Ag^+$, $Ag^0$), reaction medium (aqueous solution, soil), and exposure time.

독창적 아이디어에서 창조적 혁신까지 : 인공씨감자 기술혁신 성공사례 분석

  • 현재호
    • Proceedings of the Technology Innovation Conference
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    • 1997.07a
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    • pp.222-223
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    • 1997
  • By analyzing the successful innovation case of potato microtuber mass production technology, a representative case of technology-push type creative innovation in an imitation oriented research culture, this paper attempts to figure out conceptual model of creative innovation that is initiated by the public laboratories in catching-up country, Stages of creative innovation can be divided into the internal R&D stage and the external commercialization stage. Success of the internal R&D stage depended on autonomy to secure creative research idea and commitment of individual researchers. Psychological pressure evoked from sportlights of mass media and commitment of sponsor increased the intensity of research efforts of the researcher Recognition of research problem and its significance was intensified by site visits of agricultural fields, and the recognized higher impacts of expected research results and knowledge creation achieved were a fundamental source of self-motivation. In the stage of commercialization stage, various legal, socio-economic, and psychological barriers were confronted. In a catching-up country lacking of experiences of creative innovation, creative innovation process can be regarded as a barrier elimination and cultural revolution process. Among the barriers, psychological refusal of farmers to corn-sized potato seeds was critical, which finally enforced to further researches to enlarge the size of potato seeds. In addition, the researcher has concentrated his research efforts in one specialized research area by getting a series of similar research project funds rather than diversification. It was lucky for him to have a chance to carry out a series of similar researches in one research area during the last 10 years. In getting research funds from government and private companies continuously in one research area, both internal and external promoters played significant roles.

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Genetic Environments of Hydrothermal Vein Deposits in the Pacitan District, East Java, Indonesia (인도네시아 동부자바 빠찌딴(Pacitan) 광화대 열수 맥상 광상의 성인 연구)

  • Choi, Seon-Gyu;So, Chil-Sup;Choi, Sang-Hoon;Han, Jin-Kyun
    • Economic and Environmental Geology
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    • v.28 no.2
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    • pp.109-121
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    • 1995
  • The hydrothermal vein type deposits which comprise the Kasihan, Jompong and Gempol mineralized areas are primarily copper and zinc deposits, but they are also associated with lead and/or gold mineralization. The deposits occur within the Tertiary sedimentary and volcanic rocks in the Southern Mountain zone of the eastern Java island, Indonesia. Mineralization can be separated into two or three distinct stages (pre-and/or post- ore mineralization stages and main ore mineralization stage) which took place mainly along pre-existing fault breccia zones. The main phase of mineralization (the main ore stage) can be usually classified into three substages (early, middle and late) according to ore mineral assemblages, paragenesis, textures and their chemical compositions. Ore mineralogy and paragenesis of the three areas in the district are different from each other. Pyrite, pyrrhotite (/arsenopyrite), iron-rich (up to 20.5 mole % FeS) sphalerite and (Cu-)Pb-Bi sulfosalts are characteristic of the deposits in the Kasihan (/Jompong) area. On the other hand, pyrite + hematite + magnetite + iron-poor (2.7 to 3.6 mole % FeS) sphalerite assemblage is restricted to the Gempol area. Fluid inclusion data suggest that fluids of the main ore stage evolved from initial high temperatures (near $350^{\circ}C$) to later lower temperatures (near $200^{\circ}C$) with salinities ranging from 0.8 to 10.1 equiv. wt. percent NaCl. Each area represents a separate hydrothermal system: the mineralization at Kasihan and Jompong were largely due to early fluid boiling coupled with later cooling and dilution, whereas the mineralization at Gempol was mainly resulted from cooling and dilution by an influx of cooler meteoric waters. Fluid inclusion evidence of boiling indicates that pressures of ${\geq}95$ to 255 bars (${\geq}95$ bars for the Gempol area: $\approx$ 120 to 170 bars for the Jompong area: $\approx$ 140 to 255 bars for the Kasihan area) during portions of main ore stage mineralization. Equilibrium thermodynamic interpretation indicates that the evolution trends of the temperature versus fS2 variation of ore stage fluids in the Pacitan district follow two fashions: ore fluids at Kasihan and Jompong changed from the pyrite-pyrrhotite sulfidation stage towards pyritehematite- magnetite state, whereas those at Gempol evolved nearly along pyrite-hematite-magnetite reaction curve with decreasing temperature. The sulfur isotope compositions of sulfide minerals are consistent with an igneous source of sulfur with a ${\delta}^{34}S_{{\Sigma}s}$ value of about 3.3 per mil. The oxygen and hydrogen isotopic compositions of the fluids in each area indicate a progressive shift from the dominance of highly exchanged meteoric water at early hydrothermal systems towards an un- or less-exchanged meteoric water at later hydrothermal systems.

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The Treatment of Heavy Metal-cyanide Complexes Wastewater by Zn$^{+2}$/Fe$^{+2}$ Ion and Coprecipitation in Practical Plant (II) (아연백법 및 공침공정을 이용한 복합 중금속-시안착염 폐수의 현장처리(II))

  • Lee, Jong-Cheul;Lee, Young-Man;Kang, Ik-Joong
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.5
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    • pp.524-533
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    • 2008
  • Industrial wastewater generated in the electroplating and metal finishing industries typically contain toxic free and complex metal cyanide with various heavy metals. Alkaline chlorination, the normal treatment method destroys only free cyanide, not complex metal cyanide. A novel treatment method has been developed which destroys both free and complex metal cyanide as compared with Practical Plant(I). Prior to the removal of complex metal cyanide by Fe/Zn coprecipitation and removal of others(Cu, Ni), Chromium is reduced from the hexavalent to the trivalent form by Sodium bisulfite(NaHSO$_3$), followed by alkaline-chlorination for the cyanide destruction. The maximum removal efficiency of chromium by reduction was found to be 99.92% under pH 2.0, ORP 250 mV for 0.5 hours. The removal efficiency of complex metal cyanide was max. 98.24%(residual CN: 4.50 mg/L) in pH 9.5, 240 rpm with 3.0 $\times$ 10$^{-4}$ mol of FeSO$_4$/ZnCl$_2$ for 0.5 hours. The removal efficiency of Cu, Ni using both hydroxide and sulfide precipitation was found to be max. 99.9% as Cu in 3.0 mol of Na$_2$S and 93.86% as Ni in 4.0 mol of Na$_2$S under pH 9.0$\sim$10.0, 240 rpm for 0.5 hours. The concentration of residual CN by alkaline-chlorination was 0.21 mg/L(removal efficiencies: 95.33%) under the following conditions; 1st Oxidation : pH 10.0, ORP 350 mV, reaction time 0.5 hours, 2nd Oxidation : pH 8.0, ORP 650 mV, reaction time 0.5 hours. It is important to note that the removal of free and complex metal cyanide from the electroplating wastewater should be employed by chromium reduction, Fe/Zn coprecipitation and, sulfide precipitation, followed by alkaline-chlorination for the Korean permissible limit of wastewater discharge, where the better results could be found as compared to the preceding paper as indicated in practical treatment(I).

STUDIES ON THE EXTRACTION OF SEA WEED PROTEINS 2. Extraction of NaCl and Alcohol Soluble Proteins (해조단백질 추출에 관한 연구 2. 식염가용성 및 알콜가용성 단백질의 추출)

  • LEE Kang-Ho;RYU Hong-Soo;WOO Soon-Im
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.10 no.4
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    • pp.189-197
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    • 1977
  • In present study, the effects of various factors including the solvent concentration, extraction time and temperature, the ratio of sample vs extraction solvent, (w/v) and pH upon the extractability of the NaCl and alcohol soluble proteins of marine algae were investigated. Eight species of fresh algae, the major ones in consumption as food, namely Porphyra suborbiculata, Undarie pinnatifida, Hizikia fusiforme, Sargassum, fulvellum, Enteromorpha linza, Sargassum kjellmanianum, Codium coarctatum, and Ulva pertusa were used for the extraction of NaCl soluble protein and dried materials of four species, Perphyra suborbiculata, Undaria pinnatifida, Enteromorpha linza and Sargassum fulvellum were used for the extraction of alcohol soluble protein. The frozen and mascerated samples were prepared by the same method described in previous paper (Ryu, 1977). And the dried materials were moistened with alcohol solution before freezing. The effect of solvent concentration on the extractability of NaCl soluble protein differed from species. The extractability of Undaria Pinnatifide, Hizikia fusiforme, Perphyra suborbiculata, Enteromorpha linza, and Ulva pertusa reached maxima at 0.25M NaCl solution while the 1.0M for Sargassum fulvellum, Saygassum kjellmanianum and Codium coarctatum. In case of alcohol soluble proteins, it was shown at $20\%$ ethanol solution for Porphyra suborbiculata, Undaria pinnatifida, Enteromorpha linza, and Sargassum fulvellum. Variation of the ratio of sample vs solvent gave slight effect upon the extractability, but the ratio of 1:30(w/v) seemed most efficient for the extraction of NaCl soluble proteins and 100 ml solvent added to 1 g dried sample was effective in case of alcohol soluble proteins. Extraction time has a minimal effect upon the extraction of alcohol soluble protein, and approximately 21 to $43\%$ of algal protein was extracted within 1 hour. But in case of NaCl soluble protein extraction, the effect of time revealed differently from species to species resulting in that the extraction for 1 hour gave a maximum extractability in Ulva pertusa and Enteromorpha linza, 2 hours in Porphyra suborbiculata, Codium coarctatum and 3 hours in Undaria pinnatifica, Hizikia susiforme, Sargassum fulvellum and Sargassum kjellmanianum. When the NaCl soluble protein of Undaria pinnatifida and Enteromopha linza was extracted at various temperature, the most effective extraction temperature was $40^{\circ}C$ while the temperature was $50^{\circ}C$ for Undaria pinnatifida and $60^{\circ}C$ for Hixikia fusiforme, Sargassum fulvellum, Sargassum kjellmanianum and Codium coarctatum. Bus in case of alcohol soluble extraction, the optimum temperature was $30^{\circ}C$ for Enteromorpha linza and $40^{\circ}C$ for Undaria pinnatifida, Sargassum fulvellum and Porphyra suborbiculata. In the effect of pH on extractability, the maximum extractability of NaCl soluble proteins was obtained at pH 7to 8 and pH 8 to 9 for alcohol soluble protein.

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Studies on the Removal of Phytate from Korean Rapeseed(Brassica napus, L) Proteins -Ⅰ. Effects of pH and Salts on Protein and Phytate Solubility of Defatted Rapeseed Flour- (한국산 평지 종실 단백질의 Phytate 제거에 관한 연구 -제1보. 평지 종실 단백질과 Phytate 의 용해도에 대한 pH와 염류의 영향-)

  • Huh, Chai-Ok;Yang, Cha-Bum
    • Applied Biological Chemistry
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    • v.29 no.2
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    • pp.212-218
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    • 1986
  • Proteins in Korean rapeseeds, as in many other plantseeds, are usually bound to phytate molecules. These phytate-bound proteins are of little value as foodstuffs because of their poor solubility in digestive systems. Therefore it is necessary to remove phytates from proteins in order to convert these proteins io a useful foodstuff. In the work, an efficient procedure for removal of phytates from defatted Korean rapeseed was found. The influence of pH on the solubility of protein and phytate of rapeseed flour showed that the former was the lowest at pH 5.0 and began to increase as pH further raised. Meanwhile, the latter was the highest at pH 6.0, however, it was decreased abruptly at alkaline pH, especially to content of 1.3% at pH 11.5. The solubility cf protein was relatively high in NaCl aqueous solution at $pH\;6.0{\sim}8.0$, and did not male any noticeable difference depending on NaCl concentration. On the other hand, the solubility of phytate was high at pH of below 6.0 showing an abrupt decrease at pH of above 6.0. The solubility of protein in $CaCl_2$ aqueous solution was highest at $pH\;6.0{\sim}8.0$, however, there was no significant change at the whole range of tested pH of the solution. A maximum solubility of phytate was shown at $pH\;3.0{\sim}4.0$. And it was decreased abruptly at a higher pH of the above range and also decreased at a lower pH with higher $CaCl_2$ concentration. The solubility of phytate in $Na_2SO_3$ aqueous solution was highest at $pH\;5.0{\sim}8.0$. As the concentration goes up the maximum value of solubility was found to move to higher pHs. Depending on the concentration of $Na_2SO_3$, the decreasing pattern was changed in an alkaline solution. The solubility of phytate in the solution containing low concentration of $Ca^{2+}$ ion was low in all treatments at pH of above 7.0 and showed the maximum value at low pH as $Ca^{2+}$ ion concentration increases. The solubility of protein at pH 11.5 showed the highest value in $1mM\;Ca^{2+}$ ion solution.

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Comparison of the Uptakes of $^{99m}Tc-sestamibi\;and\;^{99m}Tc-tetrofosmin$ in Cancer Cell Lines Expressing Multidrug Resistance (다약제내성 발현 암세포에서 $^{99m}Tc-sestamibi$$^{99m}Tc-tetrofosmin$ 섭취의 비교)

  • Yoo, Jeong-Ah;Chung, Shin-Young;Seo, Myeng-Rang;Kwak, Dong-Suk;Ahn, Byeong-Cheol;Lee, Kyu-Bo;Lee, Jae-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.3
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    • pp.178-189
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    • 2003
  • Purpose: Cellular uptakes of $^{99m}Tc-sestamibi(MIBI)\;and\;\;^{99m}Tc-tetrofosmin$ into cancer cell lines expressing multidrug resistance(MDR) were investigated and compared. The effects of verapamil and cyclosporin A, well-known multidrug resistant reversing agents, on cellular uptakes of both tracers were also compared. Materials and Methods: Doxorubicin-resistant HCT15/CL02 human colorectal cell and doxorubicin-resistant K562(Adr) and vincristine-resistant K562(Vcr) human leukemic cells were studied. RT-PCR analysis was used for the detection of mdr1 mRNA expression. MDR-reversal effects with verapamil and cyclosporine A were evaluated at different drug concentrations after incubation with MIBI and tetrofosmin for 1, 15, 30, 45 and 60 min, using single-cell suspensions at $1{\times}10^6cells/ml$ incubated at $37^{\circ}C$. Radioactivity in supernatants and pellets were measured with gamma well counter. Results: The cellular uptakes of MIBI and tetrofosmin in K562(Adr) and K562(Vcr) were lower than those of parental K562 cell. In HCT15/CL02 cells and K562(Adr) cells, there were no significant difference in cellular uptakes of both tracers, but cellular uptake of MIBI was higher than that of tetrofosmin in K562(Vcr) cells. Coincubation with verapamil resulted in a increase In cellular uptakes of MIBI and tetrofosmin. Verapamil increased cellular uptakes of MIBI and tetrofosmin by HCT15/CL02 cell by 11.9- and 6.8-fold, by K562(Adr) cell by 14.3- and 8-fold and by K562(Vcr) cell by 7- and 5.7-fold in maximum, respectively. Cyciosporin A increased cellular uptakes of MIBI and tetrofosmin by HCT15/CL02 cell by 10- and 2.4-fold, by K562(Adr) cell by 44- and 13-fold and by K562(Vcr) cell by 18.8- and 11.8-fold in maximum, respectively Conclusion: Taking together, MIBI and tetrofosmin are considered as suitable radiopharmaceuticals for defecting multidrug resistance. However, MIBI seems to be a better tracer than tetrofosmin for evaluating MDR reversal effect of the modulators. Since cellular uptakes of both tracers might differ in different cell types, further experiments regarding differences in cellular uptakes between cell types should be explored.

Protective Effects of a Herbal Composition (HemoHIM) Against Apoptosis Induced by Oxidative Stress of Hydrogen Peroxide (과산화수소의 산화적 스트레스로 유도된 Apoptosis에 대한 생약복합조성물(HemoHIM)의 방호효과 평가)

  • Shin, Sung-Hae;Kim, Do-Soon;Kim, Mi-Jung;Kim, Sung-Ho;Jo, Sung-Kee;Byun, Mung-Woo;Yee, Sung-Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.9
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    • pp.1127-1132
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    • 2006
  • In our previous study, a novel herb mixture (HIM-I) of Angelica gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix was developed to protect the intestinal and immune systems and promote its recovery against radiation damage. A new herbal composition (HemoHIM) with the high immune modulating activity was developed from HIM-I. HIM-I was fractionated into ethanol fraction (HIM-I-E) and polysaccharide fraction (HIM-I-P). And HemoHIM was prepared by adding HIM-I-P to HIM-I. HemoHIM showed more effective than HIM-I in immune modulation as well as radioprotection. The present study is designed to investigate the protective effects of HIM-I, HIM-I-P, and HemoHIM on hydrogen peroxide $(H_2O_2)$ induced apoptosis of human promyelocytic leukemia (HL-60) cells. It was shown that $H_2O_2$ treatment reduced the viability of cells, and increased appearance of DNA ladders, hypodiploid (subG1) cells, and phosphatidylserine translocation level. Pretreatment of HemoHIM significantly reduced the cytotoxic effect induced by $H_2O_2$, associated with reducing the translocation of phosphatidylserine, hypodiploid cells and DNA ladders. HemoHIM appeared to be more protective than HIM-I against $H_2O_2$ induced apoptosis whereas, it exhibited similar activity to HIM-I-P. These results indicated that HemoHIM might be an useful agent for protection against oxidative stress $(H_2O_2)-induced$ apoptosis as well as immune modulation, especially since it is a relatively nontoxic natural product.