• Title/Summary/Keyword: Mitochondrial fraction

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The Relationship between F-18-FDG Uptake, Hexokinase Activity and Glut-1 Expression in Various Human Cancer Cell Lines (다양한 사람 종양세포주에서 F-18-FDG의 섭취와 Hexokinase 활성 및 Glut-1 발현과의 상관관계)

  • Kim, Bo-Kwang;Chung, June-Key;Lee, Yong-Jin;Choi, Yong-Woon;Jeong, Jae-Min;Lee, Dong-Soo;Lee, Myung-Chul
    • The Korean Journal of Nuclear Medicine
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    • v.34 no.4
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    • pp.294-302
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    • 2000
  • Purpose: To investigate the mechanisms related to F-18-FDG uptake by tumors, F-18-FDG accumulation was compared with glucose transporter-1 (Glut-1) expression and hexokinase activity in various human cancer cell lines. Materials and Methods: Human colon cancer (SNU-C2A, SNU-C4, SNU-C5), hepatocellular carcinoma (SNU-387, SNU-423, SNU-449), lung cancer (NCI-H522, NCI-H358, NCI-H1299), uterine cervical cancer (HeLa, HeLa 229, HeLa S3) and brain tumor (A172, Hs 683) cell lines were used. After 24 hr incubation of $5{\times}10^5$ cells, 37 kBq F-18-FDG was added and the uptake by cells at 10 min was measured using a gamma counter. Hexokinase activity was measured by continuous spectrophotometric rate determination. To measure mitochondrial hexokinase activity, mitochondrial fraction was separated by a high speed centrifuge. Immunohistochemical staining of Glut-1 was performed, and graded as 0, 1, 2, or 3 according to expression. Results: There was difference among F-18-FDG uptake, total and mitochondrial hexokinase activity, and Glut-1 expression with different cancer cell lines. The correlations of F-18-FDG with total hexokinase and mitochondrial hexokinase activity were low (r=0.27 and 0.26, respectively). Glut-1 expression showed a good correlation with F-18-FDG uptake (p=0.81, p=0.0015). Previously, we reported no correlation of F-18-FDG uptake with hexokinase activity in colon cancer cell lines. Thus, when colon cancer cells were excluded, F-18-FDG uptake showed higher correlation with total hexokinase and mitochondrial hexokinase activity (r=0.81, p=0.0027 and r=0.81, p=0.0049, respectively). Conclusion: Both Glut-1 expression and hexokinase activity were contributing factors related to F-18-FDG accumulation in human cancer cell lines. The relative contribution of Glut-1 expression and hexokinase activity, however, was different among different cancer cell types.

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Bryonia alba and Its Biochemical, Pharmacological Actions and Toxicity

  • Lee, Dong Wook;Aprikian, G.V.;Sohn, Hyung-Ok
    • Journal of the Korean Society of Tobacco Science
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    • v.35 no.1
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    • pp.56-61
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    • 2013
  • Bryonia alba L. belongs to the Cucurbitaceae family and grows in Europe, Asia, America, Africa, Russia, Ukraina and Armenia. The root of Bryonia alba has been used for neuropsychical diseases, psychosis, hysteria, paralysis, epilepsy, vertigo, headache, migrain, melancholia, forgetfulness, sadness, absent mindedness, delirium, cardiovascular disease, ischemia, gastrointestinal diseases, gastric ulcer and respiratory diseases. The root of Bryonia alba contains an oxidized tetra cyclic triterpens, cucurbitaceous, polyunsaturated hydrocarbons, phospholipids, phosphatidylcholines, ethereal oils, fatty acids, a great amount of amino acids, alcohol soluble enzymes, sugar, carotene, vitamin C and E. Bryonia alba increases coronary blood-flow and the amplitude of cardiac contractions. Bryonia alba has an antistressor action and increases the working capacity. Bryonia alba activates connective tissue cells. Bryonia alba markedly increases the oxygen consumption by young and senescent rat brain, liver as well as heart mitochondrial fraction as Korean Ginseng. Bryonia alba decreases lipid peroxidation after immobilization stress. In conclusion, Bryonia alba like Ginseng used in traditional medicine came from ancient time has a good perspective administration as prophylactic and medical remedy, as remedy of lot of diseases in modern medicine.

Effect of Dexamethasone Stress on Concentrations of Zinc in Blood Plasma and in Sub-Cellular Fractions of Various Tissues of Neonatal Buffalo Calves

  • Singh, Charanbir;Singha, S.P.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1022-1025
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    • 2002
  • Effect of chemical stress of daily administration of glucocorticoid (dexamethasone @0.125 mg./calf/day) injections on plasma zinc levels, Zn status of body tissues and its distribution in sub cellular fractions, was studied in neonatal buffalo calves. Daily i/m injections of dexamethasone, starting at the completion of 1 week of age and continued till 8th week, led to a significant decline in plasma Zn concentration from 3rd week onwards, which then persisted throughout the rest of the experimental period. In control group, liver had the highest concentration of zinc, followed by heart, muscle, spleen, kidney and testis. In all these tissues, cytosolic fractions had the highest (>60%) zinc levels followed by nuclear, mitochondrial and microsomal fractions. In dexamethasone treated calves, there was a significant increase in the Zn uptake by the tissues of liver and muscle. This increase in zinc concentration was observed in all the sub cellular fractions of liver and muscle, however about 80% of this increase was in cytosolic fraction. It was concluded that glucocorticoid-induced stress caused increase in Zn levels of liver/muscles and decrease in blood plasma zinc, thus indicating a redistribution of Zn in body.

Expression of Recombinant Human Cytochrome P450 1A2 in Escherichia coli Bacterial Mutagenicity Tester Strain

  • Chun, Young-Jin
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.305-309
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    • 1998
  • Human cytochrome P450 1A2 is one of the major cytochrome P450s in human liver. It is known to be capable of activating a number of carcinogens such as arylamines and heterocyclic amines. In order to develop the new bacterial mutagenicity test system with human P450, a full length of human P450 1A2 cDNA inserted into pCW bacterial expression vector was introduced to Escherichia coli WP2 uvrA strain which is a well-known E. coli strain for bacterial reverse mutagenicity assay. Expressed human P450 1A2 showed typical P450 hemoprotein spectra. Maximum expression was achieved at 48 hrs after incubating at $30^{\circ}C$ in terrific broth containing ampicillin, IPTG and other supplements. High level expression of P450 1A2 in E. coli WP2 uvrA membranes was determined in SDS-PAGE. The well-known mutagens 2-aminoanthracene and MElQ increased the revertant colonies of E. coli WP2 uvrA expressing human P450 1A2 without an exogenous rat hepatic post-mitochondrial supernatant (S9 fraction) in a dose-dependent manner. The results show that the functional expression of human P450 in bacterial mutagenicity tester strain will provide a useful tool for studying the mechanism of the mutagenesis and carcinogenesis of new drugs and environmental chemicals.

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Studies on the Metabolism of Sinigrin in Rat (흰쥐에서 sinigrin 대사에 관한 연구)

  • Huh, Keun;Shin, Uk-Seob;Lee, Sang-Il;Song, Min-Ik
    • Biomolecules & Therapeutics
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    • v.2 no.1
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    • pp.11-15
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    • 1994
  • The detoxifying properties of cruciferous vegetables components have been the subject of several recent investigations. Evidences from many biochemical and pharmacological studies indicated that higher consumption of cruciferous vegetables is associated with lower incidence of harmful actions such as hepatotoxicity and oxidative stress in animal and human populations. Recently, it has been reported that drug metabolizing and detoxifying enzyme activities were increased by cruciferous vegetable extract in which sinigrin is known to be a main active component, accounting for about 2 to 3 percents of total extract. The detoxifying effect of sinigrin has been well reported in several literatures. The metabolism of sinigrin in animal, however, has not been reported yet. That led us to study the metabolism of sinigrin in rat. Sinigrin is nown to be metabolized into three compounds, i.e., allyl isothiocyanate, glucose and potassium phosphate in cruciferous vegetables. Allyl isothiocyanate was formed in rat hepatic mitochondrial fraction in dose and incubation time dependent manner, that was confirmed by HPLC. Glucose formation was came up with results similar to that of allyl isothiocyanate. Three hours after i.p. administration of sinigrin to rat, allyl isothiocyanate appeared in rat liver, and five hours later it was detected in liver and blood. The above results suggested that sinigrin might be metabolized into allyl isothiocyanate, glucose and potassium phosphate in rat.

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Mutagenicity of Lithium Carbonate Assessed by Bacterial Reverse Mutation(Ames) Test (미생물복귀돌연변이(Ames)시험을 통한 탄산리튬의 변이원성 고찰)

  • Rim, Kyung-Taek;Kim, Soo-Jin
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.24 no.3
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    • pp.330-335
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    • 2014
  • Objectives: To evaluate the mutagenicity of lithium carbonate, a bacterial reverse mutation(Ames) test was carried out using four strains of S. typhimurium(TA1535; TA1537; TA98; and TA100) and one strain of E. coli(WP2uvrA). Materials: This was carried out in a dose range from 312.5 to $5,000{\mu}g/plate$ in triplicate with and without S9 activation, which is the most commonly used metabolic activation system supplemented by a post-mitochondrial fraction prepared from the livers of rodents treated with enzyme-inducing agents such as Aroclor 1254 or a combination of phenobarbitone and ${\beta}$-naphthoflavone. Results: No significant increases in the number of revertants were observed under the conditions examined in this study. Conclusions: Based on the above observations, it can be concluded that lithium carbonate has no mutagenic activity. Despite the results, it can have an effect by inducing acute oral toxicity, eye irritation and acute aquatic toxicity. Based on this study, we suggest that future studies should be directed toward chronic, carcinogenic testing and other related areas.

Ethyl acetate fraction of GGEx18 modulates fatty acid β-oxidizing enzymes (In vitro 동물세포에서 GGEx18의 ethyl acetate 분획물에 의한 지방산 β-산화효소 유전자 발현의 조절)

  • Joo, Byung-Soo;Lee, Hee-Young;Lee, Hye-Rim;Yoon, Mi-Chung;Seo, Bu-Il;Kim, Beom-Hoi;Shin, Soon-Shik
    • The Korea Journal of Herbology
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    • v.27 no.2
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    • pp.53-59
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    • 2012
  • Objectives : This study was undertaken to investigate the effects of the GGEx18 ethyl acetate fraction (EF) on lipid accumulation and gene expression of fatty acid-oxidizing enzymes using 3T3-L1 adipocytes, C2C12 skeletal muscle cells, and NMu2Li liver cells. Methods : PPAR${\alpha}$, AMPK and UCPs transactivation was examined in NMu2Li hepatocytes, C2C12 myocytes, and 3T3-L1 preadipocytes using transient transfection assays. Results : 1. Compared with control, EF significantly increased the mRNA expression of VLCAD in 3T3-L1 adipocytes. 2. Compared with control, EF (0.1 ${\mu}g/ml$) significantly inhibited lipid accumulation in 3T3-L1 adipocytes. 3. EF significantly increased the mRNA expression of AMPK${\alpha}$1, AMPK${\alpha}$2 and PPAR${\alpha}$ in C2C12 skeletal muscle cells compared with control. 4. EF significantly increased the mRNA expression of genes involved in fatty acid ${\beta}$-oxidation, such as thiolase, MCAD, and CPT-1 in C2C12 skeletal muscle cells compared with control. 5. EF significantly increased the mRNA expression of UCP2 involved in energy expenditure in C2C12 skeletal muscle cells compared with control. 6. Compared with control, EF (10 ${\mu}g/ml$) significantly inhibited lipid accumulation in C2C12 skeletal muscle cells. 7. EF (10 ${\mu}g/ml$) significantly increased the mRNA expression of ACOX, HD, VLCAD and MCAD in NMu2Li liver cells compared with control. Conclusions : These results suggest that EF may prevent obesity by increasing the mRNA expression of mitochondrial fatty acid ${\beta}$-oxidizing enzymes in 3T3-L1 adipocytes, by not only regulating the fatty acid oxidation through activation of AMPK and PPAR${\alpha}$, but also increasing the UCP2 mRNA expression in C2C12 skeletal muscle cells, and by stimulating the mRNA expression of fatty acid-oxidizing enzymes in NMu2Li liver cells.

Pro-apoptotic and Anti-adipogenic Effects of Proso Millet (Panicum miliaceum) Grains on 3T3-L1 Preadipocytes (기장(Panicum miliaceum)의 마우스 3T3-L1 세포에 대한 에폽토시스 유발 및 지방세포형성 억제 효능)

  • Jun, Do Youn;Lee, Ji Young;Han, Cho Rong;Kim, Kwan-Pil;Seo, Myung Chul;Nam, Min Hee;Kim, Young Ho
    • Journal of Life Science
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    • v.24 no.5
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    • pp.505-514
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    • 2014
  • To examine the anti-obese activity of miscellaneous cereal grains, 80% ethanol extracts from eight selected miscellaneous cereal grains were compared for their cytotoxic effects on 3T3-L1 murine preadipocytes. The ethanol extract of proso millet exhibited the highest cytotoxicity. Further fractionation of the ethanol extract with methylene chloride, ethyl acetate, and n-butanol showed that the cytotoxicity of the ethanol extract was mainly partitioned into the butanol fraction. As compared with differentiated mature adipocytes, 3T3-L1 preadipocytes were more susceptible to the cyctotoxicity of the butanol fraction. When each organic solvent fraction (25 ${\mu}g/ml$) was added during the differentiation period for 6 days, the cell viability was not affected significantly except for the butanol fraction, but the intracellular lipid accumulation declined to a level of 81.5%~50.3% of the control. The Oil Red O staining data also demonstrated that the ethanol extract as well as the butanol fraction could inhibit the differentiation of 3T3-L1 preadipocytes into mature adipocytes. The presence of the butanol extract during the induced adipocytic differentiation also resulted in a significant reduction in the expression levels of critical adipogenesis mediators $(C/EBP{\alpha}$, $PPAR{\gamma}$, aP2, and LPL) to a barely detectable or undetectable level and the cells retained the fibroblast-like morphology of 3T3-L1. In 3T3-L1 cells, the cytotoxicity of the butanol fraction (50-100 ${\mu}g/ml$) was accompanied by mitochondrial membrane potential (${\Delta}{\psi}m$) loss, caspase-3 activation, and PARP degradation. Taken together, these results indicate that proso millet grains possess pro-apoptotic and anti-adipocytic activities toward adipocytes, which can be applicable to prevention of obesity.

Study of Sophorae Radix on $H_2O_2$-mediated Apoptosis and Total Protein Expression Pattern in Vascular Smooth Muscle Cells (고삼이 $H_2O_2$에 의한 대동맥 평활근세포 고사 및 전체 단백질 발현에 미치는 영향)

  • Jeon In Cheol;Jeong Jae Eun;Son In Hwan;Lee Ju Seok;Jeong Seung Won;Jang Jae Ho;Lee Seon U;Lee In;Moon Byun Soon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.6
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    • pp.1652-1660
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    • 2004
  • Apoptosis of vascular smooth muscle cells(VSMCs) is essential in atherogenesis, being a factor that modulates its early progression rather than a terminal event in the course of the disease. Various stimuli, including oxide lipoproteins, altered hemodynamic stress and free radical, can induced VSMCs apoptosis in vitro. The protective effects of Sophorae Radix (SR) on apoptotic cell death induced by H₂O₂ were investigated in VSMCs. The viability of VSMCs was markedly decreased by H₂O₂. Sophorae Radix protected the H202-induced apoptotic death of VSMCs, which was characterized as nuclear fragmentation and increase of sub-G0/G1 fraction .. Sophorae Radix decreased the activation of caspase-3 like protease induced by H₂O₂ and recovered control level from H202-induced PARP, Bak, Bcl-XL and mitochondrial membrane potential. These results suggest that Sophorae Radix protected VSMCs apoptotic death induced by H₂O₂ via inactivation of caspase-3 and modulation of mitochondrial function. Also, the expression profile of proteins by using two-dimensional (2-D) gel electrophoresis was screened. Future investigations will need to explore the use of an anti atherosclerotic therapy of Sophorae Radix, which relies on inhibition of the proapoptotic activation of the vascular smooth muscle cells.

A Study on the Antilipidperoxidative Effects of Brazilin and Hematoxylin(I) (천연색소 Brazilin 및 Hematoxylin의 항지질 과산화활성에 관한 연구 (I))

  • 문창규;하배진;이수환;목명수
    • Journal of Food Hygiene and Safety
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    • v.2 no.1
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    • pp.35-40
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    • 1987
  • The antilipidperoxidative effects of Brazilin and Hematoxylin were investigated at the levels of liver~total homogenates, -microsomal fraction, -mitochondrial fraction and the sera of SD-rats intoxicated with $CCl_4$ and ethanol. Both natural dyes markedly inhibited the lipidperoxidation induced by $CCl_4$ and ethanol. Brazilin and Hematoxylin showed the inhibitory effects on the both enzymatic (NADPH-dependent) and nonenzymatic (Ascorbate-induced) lipidperoxidation pathways. but it is supposed that the antilipidperoxidative powers of them mainly result from the inhibition of the nonenzymatic lipidperoxidation.

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