• Title/Summary/Keyword: In vitro metabolism

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Biotransformation of a Fungicide Ethaboxam by Soil Fungus Cunninghamella elegans

  • PARK, MI-KYUNG;KWANG-HYEON LIU;YOONGHO LIM;YOUN-HYUNG LEE;HOR-GIL HUR;JEONG-HAN KIM
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.43-49
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    • 2003
  • Metabolism of a new fungicide ethaboxam by soil fungi was studied. Among the fungi tested, Cunninghamelia elegans produced metabolites from ethaboxam, which were not found in the control experiments. M5, a major metabolite from ethaboxam was firmly identified as N-deethylated ethaboxam by LC/MS/MS and NMR. N-Deethylated ethaboxam has been found as a single metabolite in in vitro metabolism with rat liver microsomes. Ml was proved to be 4-ethyl-2-(ethylamino)-1,3-thiazole-5-carboxamide (ETC) by comparing with the authentic compound. In addition, M2, M3, and M4, and M6 were tentatively Identified by LC/MS/MS as hydroxylated and methoxylated ethaboxams, respectively. Production of the major metabolite, N-deethylated ethaboxam, by the fungus suggested that C. elegans would be an efficient eukaryotic microbial candidate for evaluating xenobiotic-driven mammalian risk assessment.

Some Enzymes of Tricarboxylic Acid Cycle and Metabolites of Carbohydrate Metabolism in Adult Isoparorchis hypselobagri(Digenea: Trematoda) During in vitro Starvation

  • Bera, Bireshwar;Manna, Buddhadeb
    • International Journal of Industrial Entomology and Biomaterials
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    • v.18 no.2
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    • pp.91-95
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    • 2009
  • The presence of considerable amount of enzymes of TCA cycle isocitrate dehydrogenase (ICDH-NADP+, EC1.1.1.42), $\alpha$-ketogluterate dehydrogenase ($\alpha$-KGD, EC1.2.4.2) and malate dehydrogenase (MDH, EC1.1.1.37) in fresh control and in vitro starved adult Isoparorchis hypselobagri establish the functional TCA cycle in this fluke. The major metabolic end products are pyruvate, lactate, oxaloacetate and malate. The ratio of oxaloacetate/malate assess that oxaloacetate is reduced to malate and in this fluke the reverse TCA cycle is active. The pyruvate/lactate ratio shows pyruvate is reduced to lactate and the fluke is homolactate farmenters.

In vitro inhibition of 10-formyltetrahydrofolate dehydrogenase activity by acetaldehyde

  • Mun, Ju-Ae;Doh, Eun-Jin;Min, Hye-Sun
    • Nutrition Research and Practice
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    • v.2 no.4
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    • pp.195-199
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    • 2008
  • Alcoholism has been associated with folate deficiency in humans and laboratory animals. Previous study showed that ethanol feeding reduces the dehydrogenase and hydrolase activity of 10-formyltetrahydrofolate dehydrogenase (FDH) in rat liver. Hepatic ethanol metabolism generates acetaldehyde and acetate. The mechanisms by which ethanol and its metabolites produce toxicity within the liver cells are unknown. We purified FDH from rat liver and investigated the effect of ethanol, acetaldehyde and acetate on the enzyme in vitro. Hepatic FDH activity was not reduced by ethanol or acetate directly. However, acetaldehyde was observed to reduce the dehydrogenase activity of FDH in a dose- and time-dependent manner with an apparent $IC_{50}$ of 4 mM, while the hydrolase activity of FDH was not affected by acetaldehyde in vitro. These results suggest that the inhibition of hepatic FDH dehydrogenase activity induced by acetadehyde may play a role in ethanol toxicity.

Three-step in vitro digestion model for evaluating and predicting fecal odor emission from growing pigs with different dietary protein intakes

  • Lo, Shih-Hua;Chen, Ching-Yi;Wang, Han-Tsung
    • Animal Bioscience
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    • v.35 no.10
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    • pp.1592-1605
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    • 2022
  • Objective: The objective of this study was to select an effective in vitro digestion-fermentation model to estimate the effect of decreasing dietary crude protein (CP) on odor emission during pig production and to suggest potential prediction markers through in vitro and in vivo experiments. Methods: In the in vitro experiment, three diet formulations with different CP contents (170 g/kg, 150 g/kg, and 130 g/kg) but containing the same standardized ileal digestible essential amino acids (SID-EAA) were assessed. Each diet was evaluated by two different in vitro gastric-intestinal phase digestion methods (flask and dialysis), combined with fresh pig feces-ferment inoculation. Eighteen growing barrows (31.9±1.6 kg) were divided into three groups: control diet (180 g CP/kg, without SID-EAA adjustment), 170 g CP/kg diet, and 150 g CP/kg diet for 4 weeks. Results: The in vitro digestion results indicated that in vitro digestibility was affected by the gastric-intestinal phase digestion method and dietary CP level. According to the gas kinetic and digestibility results, the dialysis method showed greater distinguishability for dietary CP level adjustment. Nitrogen-related odor compounds (NH3-N, indole, p-cresol, and skatole) were highly correlated with urease and protease activity. The feeding study indicated that both EAA-adjusted diets resulted in a lower odor emission especially in p-cresol and skatole. Both protease and urease activity in feces were also closely related to odor emissions from nitrogen metabolism compounds. Conclusion: Dialysis digestion in the gastric-intestinal phase followed by fresh fecal inoculation fermentation is suitable for in vitro diet evaluation. The enzyme activity in the fermentation and the fecal samples might provide a simple and effective estimation tool for nitrogen-related odor emission prediction in both in vitro and in vivo experiments.

Cytotoxic Triterpenoid from Rubus coreanus Miq

  • Lee, Dae-Young;Kim, Dong-Hyun;Bang, Myun-Ho;Song, Myoung-Chong;Kwak, Ho-Young;Yoo, Ki-Hyun;Chung, In-Sik;Kim, Kyong-Tai;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.275-280
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    • 2007
  • Dried unripe fruits of Rubus coreanus Miq. were extracted with 80% aqueous MeOH and the concentrated extract was partitioned with EtOAc and $H_2O$. From the EtOAc fraction, four triterpenoids were isolated through repeated silica gel, ODS and Sephadex LH-20 column chromatographies. From the result of physico-chemical data including NMR, MS aud IR, the chemical structures of the compounds were determined as tormentic acid (1), myrianthic acid (2), hovenic acid (3) and 2${\alpha}$,3${\beta}$,19${\beta}$,23-tetrahydroxylolean-12-en-28-oic acid (4). Compounds 3 and 4 were isolated for the first time from this plant. All isolated compounds were evaluated for cytotoxic activity against human colon carcinoma cells using in vitro three-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, compound 3 showed a higher cytotoxicity ($IC_{50}$ = 7.8 ${\mu}M$) than doxorubicin ($IC_{50}$= 50 ${\mu}M$).

PULMONARY XENOBIOTIC CONJUGATION IN THE ISOLATED PURFUSED RABBIT LUNG AND IN VITRO: EFFECT OF ETHANOL

  • Yang, C.Mierha;Carlson, Gary P.
    • Toxicological Research
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    • v.7 no.2
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    • pp.191-208
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    • 1991
  • Pulmonary conjugation pathways may be important for the metabolism of xenobiotics introduced via airways of systemically. The objective of this study was to determine the pulmonary conjugating capacity in both the isolated perfused rabbit lung (IPRL) and in vitro, and the ability of ethanol to alter the above. The IPRL was capable of conjugating glutathione (GSH) with either 1-chloro-2,4-dinitrobenzene (CDNB) of 1,2-epoxy-(p-nitrophenoxy) propane(ENP). The pulmonary GSH conjugation with ENP was inhibited by cibacron blue, indicating the presence of glutathione-S-transferase (GST) u and/or classes, but it was not altered by buthionine sulfoximine, a selective inhibitor of Gamma-glutamylcysteine synthetase.

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Alpha-Linolenic Acid: It Contribute Regulation of Fertilization Capacity and Subsequent Development by Promoting of Cumulus Expansion during Maturation

  • Lee, Ji-Eun;Hwangbo, Yong;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Development and Reproduction
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    • v.22 no.4
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    • pp.297-307
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    • 2018
  • The objective of this study was to evaluate the effects of alpha-linolenic acid (ALA) during in vitro maturation (IVM) on cumulus expansion, nuclear maturation, fertilization capacity and subsequent development in porcine oocytes. The oocytes were incubated with 0, 25, 50, and $100{\mu}M$ ALA. Cumulus expansion was measured at 22 h, and gene expresison and nuclear maturation were analyzed at 44 h after maturation. Then, mature oocytes with ALA were inseminated, and fertilization parameters and embryo development were evaluated. In results, both of cumulus expansion and nuclear maturation were increased in $50{\mu}M$ ALA groups compared to control groups (p<0.05). However, expression of gap junction protein alpha 1 (GJA1, cumulus expansion-related gene), delta-6 desaturase (FADS1, fatty acid metabolism-related gene), and delta-5 desaturase (FADS2) mRNA in cumulus cells were reduced by $50{\mu}M$ ALA treatment (p<0.05). Cleavage rate was enhanced in 25 and $50{\mu}M$ ALA groups (p<0.05), especially, treatment of $50{\mu}M$ ALA promoted early embryo develop to 4 and 8 cell stages (p<0.05). However, blastocyst formation and number of cells in blastocyst were not differ in 25 and $50{\mu}M$ ALA groups. Our findings show that ALA treatment during maturation could improve nuclear maturation, fertilization, and early embryo development through enhancing of cumulus expansion, however, fatty acid metabolism- and cumulus expansion-related genes were down-regulated. Therefore, addition of ALA during IVM of oocytes could improve fertilization and developmental competence, and further studies regarding with the mechanism of ALA metabolism are needed.

Effects of Chromium Picolinate (CrP) on Growth Performance and Carcass Characteristics of Fattening Pigs Treated With or Without Porcine Somatotropin (pST)

  • Min, J.K.;Chung, I.B.;Chae, B.J.;Shin, I.S.;Kim, W.Y.;Choi, Y.J.;Han, I.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.1
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    • pp.20-27
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    • 1997
  • Objectives of this study was to investigate effects of supplemental chromium (Cr) as CrP in growing pigs treated with pST. Seventy two Landrace pigs weighing average 60 kg were alloted to the three treatments during the 52-d experimental period: control (corn-soybean basal diet); pST treatment (4 mg/head/day); pST + CrP treatment (4 mg and 200 ppb/head/day). Upon termination of feeding trial weighing average 105 kg, thirty-six pigs randomly selected from each treatment were slaughtered to compare carcass traits. For the study of lipid metabolism, eighteen pigs were alloted to the same treatments. Adipose tissue samples from eighteen pigs were collected to investigate lipid metabolism. All treated samples with pST and pST + CrP showed improvements in daily weight gain, regardless of sex. Feed/gain ratio significantly improved in pigs treated with pST and pST + CrP. Dressing percentages were higher in pigs treated with pST and pST + Crp. Carcass grades were significantly higher in pigs treated with pST and pST + CrP. Lipolysis of adipose tissue measured in vitro was significantly increased in pigs treated with pST, lipogenesis in vitro showed opposite tendency. Even though the current data does not show synergistic effects on the above parameters when CrP and pST were supplied at the same time, but CrP supplementation tended to improve growth performance and carcass traits of pigs treated with pST.

Pregnenolone Metabolism in the Ovarian Thecal Layers of the Rainbow Trout, Oncorhynchus mykiss: in vitro Inhibitory Effects of Cyanoketone and Trilostane (무지개송어 (Oncorhynchus mykiss)의 난소내 협막층(theca layers)에서의 Pregnenolone 대사: cyanoketone과 trilostane의 저해 효과)

  • BAEK Hea-Ja;FOSTIER Alex
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.4
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    • pp.469-474
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    • 1995
  • The effects of sperific inhibitors of $3\beta-hydroxy-\Delta^5-steroid\;dehydrogenase$ $(3\beta-HSD;\;an\;enzyme\;catalyzing\;conversion\;of \Delta^5\;steroids\;to\;\Delta^4 steroids),$ cyanoketone and trilostane, on $^3H-pregnenolone$ metabolism in isolated ovarian thecal layers have been investigated in vitro. At all doses of cyanoketone $(10^{-5}\;and\;10^{-4}\;M)$ and trilostane $(10^{-5}\;and\;10^{-4}\;M)$ $(3\beta-HSD$ enzyme activity that transforms pregnenolone to $17\alpha-hydroxyprogesterone$ was inhibited in the thecal layers. Trilostane appeared to be more efficient than cyanoketone. Trilostane at doses of $10^{-8},\;10^{-7},\;10^{-6},\;and\;10^{-5},\;M/ml$ caused a dose-response inhibition of $\Delta^4$ steroids accumulation in the medium from pregnenolone, but not completely blocked the conversion of $\Delta^5\;to\;\Delta^4$ steroids.

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