• Title/Summary/Keyword: In vitro metabolism

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Effect of Whole or Steam-flaked Corn Based Diet on Ruminal fermentation Characteristics In Vitro and Ruminal Metabolism in Korean Native Goat In Vivo (통옥수수 및 Steam-flaked 옥수수 기초사료가 반추위미생물 발효성상과 한국재래산양 반추위대사 특성에 미치는 영향)

  • Bae, G.S.;Bae, J.H.;Yun, S.J.;Chang, M.B.;Ko, J.Y.;Ha, Jong-K.
    • Journal of Animal Science and Technology
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    • v.44 no.6
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    • pp.757-768
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    • 2002
  • These study were conducted to determine the effects of a whole or steam-flaked corn based diet on rumen microbial fermentation in vitro and ruminal metabolism in the Korean Native Goat(KNG) in vivo. The experiments consisted of two dietary treatments: control, steam-flaked corn(SFC) based diet(80%) + rice straw mixed(20%)(SFCR); 100% whole corn based diet(WC). The first experiment was conducted to investigate the effect of whole corn on ruminal metabolism in vitro for 0 to 48 h. pH values were optimally maintained during incubation time, and were not significantly different between treatments. Gas production of SFCR was significantly higher than WC(p<0.01). $NH_3$-N concentration tended to increase for WC, but not significantly different between treatments. The mean value of total volatile fatty acid concentration of WC was significantly lower than SFCR(p<0.01), but SFCR and WC linearly increased as the time of incubation approached 48 h. Mean value of acetate concentration of SFCR was significantly higher than WC(p<0.01). Propionate concentration of WC for the total incubation time was significantly higher than SFCR(p<0.01). The digestibility of dry matter was not significantly different between treatments, but SFCR was somewhat higher than WC. The second experiment was conducted to effect of whole shelled corn based diet on rumen metabolism in KNG. pH values tended to decrease through all treatments. There was not a significantly difference between treatments. Microbial protein yield of SFCR was significantly higher than WC(p<0.01). $NH_3$-N concentration of WC was significantly (p<0.01) higher than SFCR. Total VFA and propionate concentration of WC was significantly higher than SFCR(p<0.01), but acetate concentrate of WC was not significantly higher than SFCR. The mean value of total lactate concentration was significantly(p<0.01) different but the value of SFCR and WC were lower than the average concentration of acidosis. In sacco DM disappearance rate of SFC was significantly(p<0.01) higher than WC.

Assessment of Feasibility for Developing Toxicogenomics Biomarkers by comparing in vitro and in vivo Genomic Profiles Specific to Liver Toxicity Induced by Acetaminophen

  • Kang, Jin-Seok;Jeong, Youn-Kyoung;Suh, Soo-Kyung;Kim, Joo-Hwan;Lee, Woo-Sun;Lee, Eun-Mi;Shin, Ji-He;Jung, Hai-Kwan;Kim, Seung-Hee;Park, Sue-Nie
    • Molecular & Cellular Toxicology
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    • v.3 no.3
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    • pp.177-184
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    • 2007
  • As a possible feasibility of the extrapolation between in vivo and in vitro systems, we investigated the global gene expression from both mouse liver and mouse hepatic cell line treated with hepatotoxic chemical, acetaminophen (APAP), and compared between in vivo and in vitro genomic profiles. For in vivo study, mice were orally treated with APAP and sacrificed at 6 and 24 h. For in vitro study, APAP were administered to a mouse hepatic cell line, BNL CL.2 and sampling was carried out at 6 and 24 h. Hepatotoxicity was assessed by analyzing hepatic enzymes and histopathological examination (in vivo) or lactate dehydrogenase (LDH) assay and morphological examination (in vitro). Global gene expression was assessed using microarray. In high dose APAPtreated group, there was centrilobular necrosis (in vivo) and cellular toxicity with the elevation of LDH (in vitro) at 24 h. Statistical analysis of global gene expression identified that there were similar numbers of altered genes found between in vivo and in vitro at each time points. Pathway analysis identified glutathione metabolism pathway as common pathways for hepatotoxicty caused by APAP. Our results suggest it may be feasible to develop toxicogenomics biomarkers or profiles by comparing in vivo and in vitro genomic profiles specific to this hepatotoxic chemical for application to prediction of liver toxicity.

In Vitro Metabolism of a New Cardioprotective Agent, KR-33028 in the Human Liver Microsomes and Cryopreserved Human Hepatocytes

  • Kim Hyojin;Yoon Yune-Jung;Kim Hyunmi;Cha Eun-Young;Lee Hye Suk;Kim Jeong-Han;Yi Kyu Yang;Lee Sunkyung;Cheon Hyae Gyeong;Yoo Sung-Eun;Lee Sang-Seop;Shin Jae-Gook;Liu Kwang-Hyeon
    • Archives of Pharmacal Research
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    • v.28 no.11
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    • pp.1287-1292
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    • 2005
  • KR-33028 (N-[4-cyano-benzo[b]thiophene-2-carbonyl]guanidine) is a new cardioprotective agent for preventing ischemia-reperfusion injury. This study was performed to identify the metabolic pathway of KR-33028 in human liver microsomes and to compare its metabolism with that of cryopreserved human hepatocytes. Human liver microsomal incubation of KR-33028 in the presence of NADPH and UDPGA resulted in the formation of four metabolites, M1, M2, M3, and M4. M1 and M2 were identified as 5-hydroxy-KR-33028 and 7-hydroxy-KR-33028, respectively, on the basis of LC/MS/MS analysis with the synthesized authentic standard. M3 and M4 were suggested to be dihydroxy-KR-33028 and hydroxy-KR-33028-glucuronide, respectively. Metabolism of KR-33028 in cryopreserved human hepatocytes resulted in the formation of M1, M2, and M4. These data show a good correlation between major metabolites formed in human liver microsomes and cryopreserved human hepatocytes. In addition, KR­33028 was found to inhibit moderately the metabolism of CYP1A2 substrates. Based on the results obtained metabolic pathway of KR-33028 is proposed.

Screening of 3-Hydroxy-3-Methylglutaryl-Coenzyme A Reductase Inhibitors In Vitro and Its Application to Pullets (HMG-CoA Reductase의 저해제 탐색과 가금의 콜레스테를 저하 효과)

  • Moon, Young-Ja;Yeom, Keum-Hee;Sung, Chang-Keun
    • The Korean Journal of Food And Nutrition
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    • v.15 no.4
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    • pp.307-313
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    • 2002
  • The primary objective of these studies was to screen the materials showing inhibitions of HMG-CoA reductase in vitro. The secondary objective was to determine the effect of garlic, lovastatin and copper on cholesterol concentrations in plasma, liver and breast tissues in pullets. The degree of inhibition of the selective samples on HMG-CoA reductase activity was determined in vitro. The inhibition ratios of water soluble garlic extracts, lovastatin (methanol extracts) and copper to HMG-CoA reductase activity were 51.3%, 87.5%, and 82.0%, respectively. Control diet (basal diet) and experimental diets, garlic powder (3% in diet), lovastatin (300mg/Kg of diet) and copper (200mg/Kg of diet) were fed to pullets in order to investigate the changes of cholesterol concentration in plasma and tissues. Total cholesterol, HDL- and LDL-cholesterol in blood plasma were significantly reduced in pullets fed diet containing 3% garlic powder. However, copper significantly increased total cholesterol compared to control and lovastatin did not affect plasma cholesterol concentration. Total cholesterol and triglyceride of liver and breast tissues in pullets were not affected by adding the cholesterol-lowering materials to diets. The data suggests that it is not easy for HMG-CoA reductase inhibitors to reduce cholesterol levels in body due to complication of cholesterol metabolism. However, garlic administration can lower the levels of plasma cholesterol in pullets.

Calcium Absorption Accelerating Effect of Chitosnn Oligosaccharides prepared by Ultrafiltration Membrane Enzymatic Reactor (한외여과막 효소반응기를 이용하여 제조한 키토산 올리고당의 칼슘흡수 촉진효과)

  • JEON You-Jin;KIM Gyu-Hyung;PARK Pyo-Jam;KIM Se-Kwon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.3
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    • pp.247-251
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    • 1999
  • In spite of various bio-functionalities of chitosan, the effects in vivo were still ambiguous because of its low absorption on organism. Therefore, chitosan oligosaccharides (COSs) are necessary to elucidate for an efficient utilization in vivo. COSs were prepared from chitosan using ultrafiltration membrane enzymatic reactor system with MWCO (molecular weight cut-off) 3,000 Da of membrane. Calcium absorption accelerating effect using COSs was examined by two methods, in vitro and in vivo. In vitro experiment, calcium absorption by the addition of COSs in a mixture solution of calcium and phosphate was higher approximately $50\%$ than that by control. In vivo using rats, group taken the diet contained $1\%$ COSs anil calcium chloride decreased about $75\%$ of calcium content excreted from feces, and then, showed about $15\%$ increase in breaking force of femur. These results demonstrated that COSs definitely involved in calcium metabolism in vivo.

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Pharmacokinetic Interaction of Chrysin with Caffeine in Rats

  • Noh, Keumhan;Oh, Do Gyeong;Nepal, Mahesh Raj;Jeong, Ki Sun;Choi, Yongjoo;Kang, Mi Jeong;Kang, Wonku;Jeong, Hye Gwang;Jeong, Tae Cheon
    • Biomolecules & Therapeutics
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    • v.24 no.4
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    • pp.446-452
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    • 2016
  • Pharmacokinetic interaction of chrysin, a flavone present in honey, propolis and herbs, with caffeine was investigated in male Sprague-Dawley rats. Because chrysin inhibited CYP1A-selective ethoxyresorufin O-deethylase and methoxyresorufin O-demethylase activities in enriched rat liver microsomes, the pharmacokinetics of caffeine, a CYP 1A substrate, was studied following an intragastric administration with 100 mg/kg chrysin. In addition to the oral bioavailability of chrysin, its phase 2 metabolites, chrysin sulfate and chrysin glucuronide, were determined in rat plasma. As results, the pharmacokinetic parameters for caffeine and its three metabolites (i.e., paraxanthine, theobromine and theophylline) were not changed following chrysin treatment in vivo, despite of its inhibitory effect on CYP 1A in vitro. The bioavailability of chrysin was found to be almost zero, because chrysin was rapidly metabolized to its sulfate and glucuronide conjugates in rats. Taken together, it was concluded that the little interaction of chrysin with caffeine might be resulted from the rapid metabolism of chrysin to its phase 2 metabolites which would not have inhibitory effects on CYP enzymes responsible for caffeine metabolism.

Effect of Heavy Metals on the Secretion of Amylase in Rat Pancreatic Fragments (중금속류가 취절편의 Amylase 분비에 미치는 영향)

  • Kim, Hea-Young;Kim, Won-Joon
    • The Korean Journal of Pharmacology
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    • v.17 no.2
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    • pp.31-36
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    • 1981
  • Heavy metals which are present as trace elements in human body have been known to modify various enzymatic reaction. These metals can be essential or non-essential. Zinc, copper and calcium are essential in maintaining some biological processes, whereas non-essential metals such as cadmium, lead and mercury produce accumulatve toxic effect. Cadmium accumulated in pancreas can cause toxicity and damage of pancreatic cells, thereby influencing CHO metabolism. Lead compounds are known to produce toxic effects on the kidney, digestive system and brain fellowed by inhibition of activity of ${\rho}-aminolevulinic$ acid and biosynthesis of hemoproteins and cytochrome. Evidence has been accumulated that zinc not only acts as a cofactor in enzyme reaction but also prevents toxic effect induced by heavy metal such as copper and cadmium. To demonstrate the effect of heavy metals on pancreatic secretion, part of uncinate pancreas was taken and incubated in Krebs-Ringer bicarbonate buffer with heavy metals used. Additional treatment with CCK-OP was performed when needed. After incubation during different period of time, medium was analyzed for amylase activity using Bernfeld's method. The present study was attempted in order to elucidate the effect of several kinds of heavy metal on exocrine pancreatic secretion in vitro. The results obtained are as follows: 1) CCK-OP stimulated significantly amylase release from pancreatic fragments in vitro. 2) CCK-OP response of amylase release from pancreatic fragments was inhibited by treatmant with cadmium, especially high doses of cadmium. 3) CCK-OP response of amylase release from pancreatic fragments was inhibited when pretreated with $10^{-4}M$ copper chloride. 4) Lead chloride at the concentration of $10^{-3}M\;and\;10^{4}M$ stimulated the basal amylase release in vitro but CCK-OP response did not augment by lead chloride. 5) Zine chloride did not affect amylase release from pancreatic fragment in vitro. From the results mentioned above, it is suggested that CCK-OP response was inhibited it the amylase release from pancreatic fragments pretreated with cadmium and copper chloride.

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RTP1, a Rat Homologue of Adenovirus ElA-associated Protein BS69, Interacts with DNA Topoisomerase II

  • Oh, Misook;Rha, Geun-Bae;Yoon, Jeong-Ho;Sunwoo, Yang-Il;Hong, Seung-Hwan;Park, Sang-Dai
    • Animal cells and systems
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    • v.6 no.3
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    • pp.277-282
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    • 2002
  • Topoisomearse II is an essential enzyme in all organisms with several independent roles in DNA metabolism. Recently, it has been demonstrated that the C-terminal region of topoisomerases II is associated with hetero-logous protein-protein interactions in human and yeast. In this study, we identified that RTP1, a rat homologue of EIA binding protein BS69, is another topoisomerae II interacting protein by yeast two-hybrid screening. RTP1 has an E1A-binding domain and a MYND motif, which are known to be required for transcriptional regulation by binding to other proteins and interaction with the leucine zipper motif of topoisomerase II. The physical interaction between RTP1 and topoisomerase ll$\alpha$ was examined by GST pull-down assay in vitro. The expression level of RTP1 peaks in S phase as that of topoisomerase ll$\alpha$. These results suggest that the interaction between topoisomerase ll$\alpha$ and RTP1 might play an important role in regulating the transcription of genes involved in DNA metabolism in higher eukaryotes.

Measurement of Inhibitory Activities on 3-Hydorxy-3-Methylglutaryl CoA Reductase and Acyl-CoA:Cholesterol Acyltransferase by Various Plant Extracts in vitro (시험관법에의한 식물열수추출물의 3-Hydroxy-3-Methylglutaryl CoA Reductase 및 Acyl-CoA:Cholesterol Acyltransferase 활성 저해도 측정)

  • 최명숙;이희자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.4
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    • pp.958-962
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    • 1999
  • Hydroxy methylglutaryl CoA(HMG CoA) reductase and acyl CoA:cholesterol acyltransferase(ACAT) are two important enzymes that are associated with regulation of cholesterol metabolism. The inhibitors of HMG CoA reductase and ACAT are very effective in lowering serum cholesterol in most animal species. In present study, various plant extracts with hot water were used to examine the inhibitory activities against HMG CoA reductase and ACAT that are involved in cholesterol biosynthesis and cholesterol esterification in tissues, respectively. The extracts of Fagophyrum rotundatum, Rosa multiflora, Rosa rugosa and Alisma orientalis exhibited significant inhibitory activities against the ACAT, 29%, 24%, 19%, and 18%, respectively. However the extracts of Typha augustifolia, Polygonum cuspidatum, Crataegus pinnatifida, Polygonum multiflorum inhibited the HMG CoA reductase activity by 53%, 42%, 37%, and 33% respectively. Results suggest that these plant extracts might play important roles in the regulation of the cholesterol metabolism in vivo.

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Osteoclast Differentiation Factor Engages the PI 3-kinase, p38, and ERK pathways for Avian Osteoclast Differentiation

  • Kim, Hong-Hee;Kim, Hyun-Man;Kwack, Kyu-Bum;Kim, Si-Wouk;Lee, Zang-Hee
    • BMB Reports
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    • v.34 no.5
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    • pp.421-427
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    • 2001
  • Osteoclasts, cells primarily involved in bone resorption, originate from the hematopoietic precursor cells of the monocyte/macrophage lineage and differentiate into multinucleated mature forms. We developed an in vitro osteoclast culture system using embryonic chicken bone marrow cells. This culture system can be utilized in studies on the differentiation and function of osteoclasts. Phosphatidylinositol 3-kinase (PI3-kinase) and mitogen-activated protein kinases (MAPKs) have been implicated in diverse cellular functions including proliferation, migration, and survival. Using the developed avian osteoclast culture system, we examined the involvement of these kinases in osteoclast differentiation by employing specific inhibitors of the kinases. We Found that the inhibition of the PI 3-kinase, p38, or ERK interfered with osteoclast formation, suggesting that the signaling pathways that involve these molecules participate in the process of chicken osteoclast differentiation.

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