• Title/Summary/Keyword: in vitro Metabolism

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Potential in vitro Protective Effect of Quercetin, Catechin, Caffeic Acid and Phytic Acid against Ethanol-Induced Oxidative Stress in SK-Hep-1 Cells

  • Lee, Ki-Mo;Kang, Hyung-Sik;Yun, Chul-Ho;Kwak, Hahn-Shik
    • Biomolecules & Therapeutics
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    • v.20 no.5
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    • pp.492-498
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    • 2012
  • Phytochemicals have been known to exhibit potent antioxidant activity. This study examined cytoprotective effects of phytochemicals including quercetin, catechin, caffeic acid, and phytic acid against oxidative damage in SK-Hep-1 cells induced by the oxidative and non-oxidative metabolism of ethanol. Exposure of the cells to excess ethanol resulted in a significant increase in cytotoxicity, reactive oxygen species (ROS) production, lipid hydroperoxide (LPO), and antioxidant enzyme activity. Excess ethanol also caused a reduction in mitochondrial membrane potential (MMP) and the quantity of reduced glutathione (GSH). Co-treatment of cells with ethanol and quercetin, catechin, caffeic acid and phytic acid significantly inhibited oxidative ethanol metabolism-induced cytotoxicity by blocking ROS production. When the cells were treated with ethanol after pretreatment of 4-methylpyrazole (4-MP), increased cytotoxicity, ROS production, antioxidant enzyme activity, and loss of MMP were observed. The addition of quercetin, catechin, caffeic acid and phytic acid to these cells showed suppression of non-oxidative ethanol metabolism-induced cytotoxicity, similar to oxidative ethanol metabolism. These results suggest that quercetin, catechin, caffeic acid and phytic acid have protective effects against ethanol metabolism-induced oxidative insult in SK-Hep-1 cells by blocking ROS production and elevating antioxidant potentials.

Metabolic Stability of [18F]Fluoroalkylbiphenyls

  • Lee, Kyo-Chul;Lee, Sang-Yoon;Choe, Yearn-Seong;Chi, Dae-Yoon
    • Bulletin of the Korean Chemical Society
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    • v.25 no.8
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    • pp.1225-1230
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    • 2004
  • The stability of fluoroalkyl groups as a pendent on the phenyl ring was measured in vitro using rat hepatic microsomes and human serum to predict their in vivo stabilities. We have prepared three [$^{18}F$]fluoroalkyl-biphenyls as the model compounds of fluoroalkyl aromatic compounds to compare the in vitro stabilities. In addition, in vitro stabilities were measured separately using rat hepatic microsomes and human serum at $37^{\circ}C$. Fluoroethylbiphenyl had similar or slightly superior stability to fluoropropylbiphenyl and these two compounds were much more stable than fluoromethylbiphenyl in vitro.

Genome of Bifidobacteria and Carbohydrate Metabolism

  • Bondue, Pauline;Delcenserie, Veronique
    • Food Science of Animal Resources
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    • v.35 no.1
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    • pp.1-9
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    • 2015
  • In recent years, the knowledge about bifidobacteria has considerably evolved thanks to recent progress in molecular biology. The analysis of the whole genome sequences of 48 taxa of bifidobacteria offers new perspectives for their classification, especially to set up limit between two species. Indeed, several species are presenting a high homology and should be reclassified. On the other hand, some subspecies are presenting a low homology and should therefore be reclassified into different species. In addition, a better knowledge of the genome of bifidobacteria allows a better understanding of the mechanisms involved in complex carbohydrate metabolism. The genome of some species of bifidobacteria from human but also from animal origin demonstrates high presence in genes involved in the metabolism of complex oligosaccharides. Those species should be further tested to confirm their potential to metabolize complex oligosaccharides in vitro and in vivo.

Effect of Scutellariae Radix Extract on Human CYP450 Mediated-Drug Metabolism

  • Yoo, Hye-Hyun;Lim, Sun-Young;Kim, Dong-Hyun
    • Journal of Pharmaceutical Investigation
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    • v.41 no.3
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    • pp.143-146
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    • 2011
  • Scutellariae Radix is widely used in the traditional herbal medicine for the treatment of fever, cough, dysentery, hepatitis and hypertension in Korea, China and Japan. In this study, we investigated the effects of 70% ethanolic extract of Scutellariae Radix (SRE) on CYP450-mediated drug metabolism in the in vitro systems using human liver microsomes and hepatocytes. The microsomal incubation assay showed that SRE inhibited the drug metabolism reactions catalyzed by CYP1A2, CYP2C8 and CYP2C9 in a dose-dependent manner. In particular, SRE was shown to strongly inhibit the metabolic activity of CYP1A2 with an $IC_{50}$ value of 4.6 ${\mu}g/mL$. When SRE was evaluated for its effect on the induction of CYP450 enzyme activities in cryopreserved human hepatocytes, SRE did not exhibit any effect.

Characterization of Deoxypodophyllotoxin Metabolism in Rat Liver Microsomes

  • Lee, Sang-Kyu;Jun, In-Hye;Kang, Mi-Jeong;Jeon, Tae-Won;Kim, Ju-Hyun;Seo, Young-Min;Shin, Sil;Choi, Jae-Ho;Jeong, Hye-Gwang;Lee, Seung-Ho;Jeong, Tae-Cheon
    • Biomolecules & Therapeutics
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    • v.16 no.3
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    • pp.190-196
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    • 2008
  • Deoxypodophyllotoxin (DPT) is a medicinal herb product isolated from Anthriscus sylvestris. DPT possesses beneficial activities in regulating immediate-type allergic reaction and anti-inflammatory activity through the dual inhibition of cyclooxygenase-2 and 5-lipoxygenase. In the present study, the metabolism of DPT was further characterized in rat liver microsomes isolated from male Sprague Dawley rats. The metabolism of DPT was NADPH-dependent. In addition, when liver microsomes were incubated with SKF-525A, a well-known CYP inhibitor, in the presence of $\beta$-NADPH, the metabolism of DPT was significantly inhibited. Using enriched rat liver microsomes, the anticipated isoforms of cytochrome P450s (CYPs) in the metabolism of DPT were partially characterized. Phenobarbital-induced microsomes increased in the formation of metabolite M1. The metabolite M3 was only produced in the enriched microsomes isolated from dexamethasone-treated rats. The results indicated that the metabolism of DPT would be CYP-dependent and that CYP2B and CYP3A might be important in the metabolism of DPT in rats.

Influence of Temperature and pH on Fermentation Pattern and Methane Production in the Rumen Simulating Fermenter (RUSITEC)

  • Bhatta, R.;Tajima, K.;Kurihara, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.376-380
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    • 2006
  • An experiment was conducted to study the effect of temperature and pH on in vitro nutrient degradability, volatile fatty acid profile and methane production. The fermenter used was the semi-continuous system, known as the rumen simulation technique (RUSITEC). Sixteen cylinders were used at one time with a volume of 800 ml, the dilution rate was set at 3.5%/hour, the infused buffer being McDougall's artificial saliva. Basal diet (9.6 g DM) used in RUSITEC consisted of (DM) 6.40 g Timothy hay, 1.86 g crushed corn and 1.34 g soybean meal. The food for the fermentation vessel was provided in nylon bags, which were gently agitated in the liquid phase. The experiment lasted for 17 d with all the samples taken during the last 5 d. Treatments were allocated at random to four vessels each and were (1) two temperature levels of $39^{\circ}C$ and $41^{\circ}C$ (2) two pH levels of 6.0 and 7.0. The total diet contained ($g\;kg^{-1}$ DM) 957 OM, 115 CP and $167MJ\;kg^{-1}$ (DM) GE. Although increase in temperature from $39^{\circ}C$ to $41^{\circ}C$ reduced degradation of major nutrients in vitro, it was non-significant. Interaction effect of temperature with pH also reflected a similar trend. However, pH showed a significant (p<0.05) negative effect on the degradability of all the nutrients in vitro. Altering the in vitro pH from 7 to 6 caused marked reduction in DMD from 60.2 to 41.8, CPD from 76.3 to 55.3 and GED from 55.3 to 35.1, respectively. Low pH (6) depressed total VFA production (61.9 vs. 34.9 mM) as well as acetate to propionate ratio in vitro (from 2.0 to 1.5) when compared to pH 7. Compared to pH 7, total gas production decreased from 1,841 ml to 1,148 ml at pH 6, $CO_2$ and $CH_4$ production also reduced from 639 to 260 ml and 138 to 45 ml, respectively. This study supported the premise that pH is one of the principal factors affecting the microbial production of volatile fatty acids and gas. Regulating the ruminal pH to increase bacterial activity may be one of the methods to optimize VFA production, reduce methane and, possibly, improve animal performance.

RNA Polymerase II Inhibitor, ${\alpha}$-Amanitin, Affects Gene Expression for Gap Junctions and Metabolic Capabilities of Cumulus Cells, but Not Oocyte, during In Vitro Mouse Oocyte Maturation

  • Park, Min-Woo;Lee, Hyun-Seo;Kim, Eun-Young;Lee, Kyung-Ah
    • Development and Reproduction
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    • v.17 no.1
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    • pp.63-72
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    • 2013
  • A specific inhibitor of RNA polymerase II, ${\alpha}$-amanitin is broadly used to block transcriptional activities in cells. Previous studies showed that ${\alpha}$-amanitin affects in vitro maturation of cumulus-oocyte-complex (COC). In this study, we evaluated the target of ${\alpha}$-amanitin, and whether it affects oocytes or cumulus cells (CCs), or both. We treated ${\alpha}$-amanitin with different time period during in vitro culture of denuded oocytes (DOs) or COCs in comparison, and observed the changes in morphology and maturation status. Although DOs did not show any change in morphology and maturation rates with ${\alpha}$-amanitin treatment, oocytes from COCs were arrested at metaphase I (MI) stage and CCs were more scattered than control groups. To discover causes of meiotic arrest and scattering of CCs, we focused on changes of cumulus expansion, gap junctions, and cellular metabolism which to be the important factors for the successful in vitro maturation of COCs. Expression of genes for cumulus expansion markers (Ptx3, Has2, and Tnfaip6) and gap junctional proteins (Gja1, Gja4, and Gjc1) decreased in ${\alpha}$-amanitin-treated CCs. However, these changes were not observed in oocytes. In addition, expression of genes related to metabolism (Prps1, Rpe, Rpia, Taldo1, and Tkt) decreased in ${\alpha}$-amanitin-treated CCs but not in oocytes. Therefore, we concluded that the transcriptional activities of CCs for supporting suitable transcripts, especially for its metabolic activities and formation of gap junctions among CCs as well as with oocytes, are important for oocytes maturation in COCs.

Changes in the Pharmacokinetics of Rosiglitazone, a CYP2C8 Substrate, When Co-Administered with Amlodipine in Rats

  • Kim, Seon-Hwa;Kim, Kyu-Bong;Um, So-Young;Oh, Yun-Nim;Chung, Myeon-Woo;Oh, Hye-Young;Choi, Ki-Hwan
    • Biomolecules & Therapeutics
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    • v.17 no.3
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    • pp.299-304
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    • 2009
  • Rosiglitazone maleate (RGM) is widely used for improving insulin resistance. RGM is a moderate inhibitor of cytochrome P450 2C8 (CYP2C8) and is also mainly metabolized by CYP2C8. The aim of this study was to determine whether the effect of RGM on CYP2C8 is altered by co-treatment with other drugs, and whether amlodipine camsylate (AC) changes the pharmacokinetics (PK) of RGM. Of the 11 drugs that are likely to be co-administered with RGM in diabetic patients, seven drugs lowered the $IC_{50}$ value of RGM on CYP2C8 by more than 80%. In vitro CYP2C8 inhibitory assays of RGM in combination with drugs of interest showed that the $IC_{50}$ of RGM was decreased by 98.9% by AC. In a pharmacokinetic study, Sprague-Dawley (SD) rats were orally administered 1 mg/kg of RGM following by single or 10-consecutive daily administrations of 1.5 mg/kg/day of AC. No significant changes in the pharmacokinetic parameters of RGM were observed after a single administration of AC, but the AUC and $C_{max}$ values of RGM were significantly reduced by 36% and 31%, respectively, by multiple administrations of AC. In conclusion, RGM was found to be affected by AC by in vitro CYP2C8 inhibition testing, and multiple dosing of AC appreciably changed the pharmacokinetics of RGM. These findings suggest that a drug interaction exists between AC and RGM.

EFFECTS OF CIMATEROL ON THE GROWTH PERFORMANCE, CARCASS CHARACTERISTICS AND TISSUE METABOLISM IN BROILER CHICKS FED DIFFERENT DIETARY ENERGY

  • Kim, Y.G.;Han, I.K.;Ha, J.K.;Choi, Y.J.;Lee, M.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.1
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    • pp.103-112
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    • 1994
  • This study was a sequential experiment consisting if feeding trial and in vitro culture studies. Feeding was conducted by $2{\times}2{\times}2$ factorial design with two cimaterol levels (0, 0.25 mg/kg), two energy levels (3,200, 2,900 ME kcal/kg) and two sexes. In starting period (0-21 days) broilers were fed diets containing two energy level without dietary supplementation of cimaterol. During finishing period (21-42 days) cimaterol groups were fed cimaterol supplemented diets. In vitro cultures were carried out to study the cellular metabolism of protein and fat in liver and adipose tissues prepared from chicks used in feeding trials. Body weight gain was significantly improved by the administration of cimaterol to experimental diets by 2.4% (p < 0.05). Feed intake was reduced by cimaterol administration at the high energy level, but this trend was reversed at low energy level. Feed efficiency was improved by cimaterol administration and at high energy level the difference (5.7%) was significant(p < 0.05). The administration of cimaterol had no effects on percentage of abdominal fat content, giblet and neck. There was little difference in carcass yield between control and cimaterol treated group. The administration of cimaterol had no effects on nutrient metabolizability or carcass composition. The results of in vitro studies with liver tissues showed that cimaterol increased the lipolytic activities (p < 0.05) and decreased lipogenic activities (p < 0.05). In in vitro studies with acinar cell of liver tissues. cimaterol increased the amount of retained protein and decreased secreted protein at high energy level. but the trend was opposite at low energy level.

Somatic Embryogenesis: Morphogenesis, Physiology, Biochemistry and Molecular Biology

  • Thorpe, Trevor A.
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.4
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    • pp.245-258
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    • 2000
  • Somatic embryogenesis has become a major tool in the study of plant embryology, as it is possible in culture to manipulate cells of many plant species to produce somatic embryos in a process that is remarkably similar to zygotic embryogenesis. Traditionally, the process has been studied by an examination of the ex vitro factors which influence embryo formation. Later structural, physiological and biochemical approaches have been applied. Host recently, molecular tools are being used. Together, these various approaches are giving valuable information on the process. This article gives an overview of somatic embryogenesis by reviewing information on the morphogenesis, physiology, biochemistry and molecular biology of the process. Topics covered include a brief description of the factors involved in the production of embryogenic cells. Carrot cell suspension is most commonly used, and the development of a high frequency and synchronous system is outlined. At the physiological and biochemical lev-els various topics, including the reactivation of the cell cycle, changes in endogenous growth regulators, amino acid, polyamine, DNA, RNA and protein metabolism, and embryogenic factors in conditioned medium are all discussed. Lastly, recent information on genes and molecular markers of the embryogenic process are outlined. Somatic embryogenesis, the best example of totipotency in plant cells, is not only an important tool in studies in basic biology, but is potentially of equal significance in the micropropagation of economically important plants.

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