• Title/Summary/Keyword: In vitro metabolism

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Role of Metabolism by Intestinal Bacteria in Arbutin-Induced Suppression of Lymphoproliferative Response in vitro

  • Kang, Mi-Jeong;Ha, Hyun-Woo;Kim, Ghee-Hwan;Lee, Sang-Kyu;Ahn, Young-Tae;Kim, Dong-Hyun;Jeong, Hye-Gwang;Jeong, Tae-Cheon
    • Biomolecules & Therapeutics
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    • v.20 no.2
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    • pp.196-200
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    • 2012
  • Role of metabolism by intestinal bacteria in arbutin-induced immunotoxicity was investigated in splenocyte cultures. Following an incubation of arbutin with 5 different intestinal bacteria for 24 hr, its aglycone hydroquinone could be produced and detected in the bacterial culture media with different amounts. Toxic effects of activated arbutin by intestinal bacteria on lymphoproliferative response were tested in splenocyte cultures from normal mice. Lipopolysaccharide and concanavalin A were used as mitogens for B- and T-cells, respectively. When bacteria cultured medium with arbutin was treated into the splenocytes for 3 days, the medium cultured with bacteria producing large amounts of hydroquinone induced suppression of lymphoproliferative responses, indicating that metabolic activation by intestinal bacteria might be required in arbutin-induced toxicity. The results indicated that the present testing system might be applied for determining the possible role of metabolism by intestinal bacteria in certain chemical-induced immunotoxicity in animal cell cultures.

A Novel in Vitro Method for the Metabolism Studies of Radiotracers Using Mouse Liver S9 Fraction (생쥐 간 S9 분획을 이용한 방사성추적자 대사물질의 새로운 체외 측정방법)

  • Ryu, Eun-Kyoung;Choe, Yearn-Seong;Kim, Dong-Hyun;Lee, Sang-Yoon;Choi, Yong;Lee, Kyung-Han;Kim, Byung-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.4
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    • pp.325-329
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    • 2004
  • Purpose: Usefulness of mouse liver S9 fraction was evaluated for the measurement of the metabolites in the in vitro metabolism study of $^{18}F$-labeled radiotracers. Materials and Methods: Mouse liver S9 fraction was isolated at au early step in the course of microsome preparation. The in vitro metabolism studies were tarried out by incubating a mixture containing the radiotracer, S9 fraction and NADPH at $37^{\ciirc}C$, and an aliquot of the mixture was analyzed at the indicated time points by radio-TLC. Metabolic defluorination was further confirmed by the incubation with calcium phosphate, a bone mimic. Results: The radiotracer $[^{18}F]1$ underwent metabolic defluorination within 15 min, which was consistent with the results of the in vivo method and the in vitro method using microsome. Radiotracer $[^{18}F]2$ was metabolized to three metabolites including $4-[^{18}F]fluorobenzoic$ acid within 60 min. It is likely that the one of these metabolites at the origin of radio-TLC was identical with the one that obtained from the in vivo and in vitro (microsome) method. Compared with the in vitro method using microsome, the method using S9 fraction gave a similar pattern of the metabolites but with a different ratio, which can be explained by the presence of cytosol in the S9 fraction. Conclusion: These results suggest that the findings of the in vitro metabolism studies using S9 fraction can reflect the in vivo metabolism of novel radiotracers in the liver. Moreover, this method can be used as a tool to determine metabolic defluorination along with calcium phosphate absorption method.

Human Tissue-Engineered Skeletal Muscle: A Tool for Metabolic Research

  • Ji-Hoon Kim;Seung-Min Yu;Jang Won Son
    • Endocrinology and Metabolism
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    • v.37 no.3
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    • pp.408-414
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    • 2022
  • Skeletal muscle is now regarded as an endocrine organ based on its secretion of myokines and exerkines, which, in response to metabolic stimuli, regulate the crosstalk between the skeletal muscle and other metabolic organs in terms of systemic energy homeostasis. This conceptual basis of skeletal muscle as a metabolically active organ has provided insights into the potential role of physical inactivity and conditions altering muscle quality and quantity in the development of multiple metabolic disorders, including insulin resistance, obesity, and diabetes. Therefore, it is important to understand human muscle physiology more deeply in relation to the pathophysiology of metabolic diseases. Since monolayer cell lines or animal models used in conventional research differ from the pathophysiological features of the human body, there is increasing need for more physiologically relevant in vitro models of human skeletal muscle. Here, we introduce recent studies on in vitro models of human skeletal muscle generated from adult myogenic progenitors or pluripotent stem cells and summarize recent progress in the development of three-dimensional (3D) bioartificial muscle, which mimics the physiological complexity of native skeletal muscle tissue in terms of maturation and functionality. We then discuss the future of skeletal muscle 3D-organoid culture technology in the field of metabolic research for studying pathological mechanisms and developing personalized therapeutic strategies.

Antigenotoxicity of Galangin and its Action Mechanism (Galangin의 유전독성 억제효과와 작용기전)

  • 허문영;류재천
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.77-82
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    • 1998
  • In order to compare the suppressive effect of galangin on the genotoxicity by N-methyl-N-nitrosourea (MNU) or benzo[a]pyrene B(a)P, in vivo micronycleus test using mouse peripheral blood and in vitro sister chromatid exchange(SCE) test using mouse spleen lymphocytes were performed. MNU or B(a)P-induced micronucleated reticulocytes in vivo was decreased by the simultaneous treatment of galangin. MNU or B(a)P-induced SCEs in vitro was also decreased by the simultaneous treatment of galangin. On the other hand, the determinations of [$^3$H]MNU-induced total DNA binding and methylated DNA were performed to find out the mechanism of action. [$^3$H]MNU-induced total DNA binding was inhibited by the treatment of galangin in calf thymus DNA. HPLC analysis of DNA hydrolysates showed that galangin caused a decrease of 7-methyl guanine and $O^{6}$-methyl guanine in calf thymus DNA. To elucidate the action mechanism of galangin against B(a)P, alteration of B(a)P metabolism was studied. Galangin inhibited B(a)P metabolism in the presence of S-9 mix and decreased B(a)P-DNA binding in calf thymus DNA with S-9 mix.

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Identification of Benzidine Metabolites in Rats by Gas Chromatography/Mass Selective Detector and its Toxicity in vitro (Gas-Chromatography/Mass Selective Detector를 사용하여 쥐의 뇨시료 중 benzidine 대사체의 확인 및 in vitro 독성)

  • 류재천;권오승
    • YAKHAK HOEJI
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    • v.44 no.5
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    • pp.384-390
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    • 2000
  • Metabolism study of the dye, benzidine, was performed by gas chromatography-mass selective detector (GC/MSD) in the urine of rats orally administered 100 mg/kg benzidine. Urine samples were collected in metabolic cages for 0-24, 24-48, and 48-72 hrs. Ten ml of the urine was extracted with XAD-2 resin and the XAD-2 column was eluted with methanol. After evaporation, benzidine and its metabolites were extracted with diethyl ether (for non-conjugated fraction). For conjugated metabolites, $\beta$-glucu-ronidase was added to the aqueous layer that was incubated for 1 hr at 5$0^{\circ}C$ and the aqueous layer was extracted as in non-conjugated fraction. Aliquot of trimethylsilylated derivatives was applied to the GC/MSD. The mutagenicity of benzidine and its acetylated metabolites was tested by histidine/reversion assay. Five metabolites observed and confirmed either by electron impact and chemical ionization modes of the GC/MSD, or authentic compounds were monoacetyl-, diacetyl-, hydroxyacetyl-, hydroxydiacetyl-, and hydroxy-benzidine. Monoacetyl-benzidine was more potent than benzidine in histidine/reversion assay. This data indicates that monoacetylation of benzidine may be one of the metabolites produced in metabolic activation process.

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Fruit and vegetable discards preserved with sodium metabisulfite as a high-moisture ingredient in total mixed ration for ruminants: effect on in vitro ruminal fermentation and in vivo metabolism

  • Ahmadi, Farhad;Lee, Won Hee;Oh, Young-Kyoon;Park, Keunkyu;Kwak, Wan Sup
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.3
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    • pp.446-455
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    • 2020
  • Objective: Our recent series of laboratory- and large-scale experiments confirmed that under aerobic and anaerobic conditions, sodium metabisulfite (SMB) was effective in preserving nutrients and antioxidant capacity of highly perishable fruit and vegetable discards (FVD). Hence, the purpose of this study was to examine how partial inclusion of SMB-treated FVD in total mixed ration (TMR) influences in vitro ruminal fermentation, whole-tract digestibility, nitrogen metabolism, blood metabolites, and voluntary feed intake of sheep. Methods: The FVD were mixed thoroughly with 6 g SMB/kg wet biomass and kept outdoors under aerobic conditions for 7 days. Four TMRs including four levels of SMB-treated FVD (as-fed basis) at 0%, 10%, 20%, and 30% (equaling to 0%, 1.9%, 3.8%, and 5.7% on dry matter basis, respectively), were prepared as replacement for corn grain. The ruminal fermentation metabolites were studied using an in vitro gas production test. Four mature male Corriedale sheep were assigned at random to the 4 diets for two separate sub-experiments; i) digestibility trial with four 21-d periods, and ii) voluntary feed intake trial with four 28-d periods. Results: Inclusion of SMB-treated FVD in the TMR tended to quadratically increase partitioning factor. No effect was seen on total-tract digestibility of organic matter, ether extract, crude protein, and acid detergent fiber, except for neutral detergent fiber digestibility that tended to linearly increase with increasing SMB-treated FVD in the TMR. The progressive increase of FVD preserved with SMB in the diet had no effect on nitrogen metabolism. Treatment had no effect on serum antioxidant capacity and blood metabolites assayed. Voluntary feed intake was not impaired by inclusion of SMB-treated FVD in the TMR. Conclusion: It appears that FVD preserved with SMB can be safely incorporated into TMR as replacement of corn grain without impairment of nutrient metabolism and feed intake.

Absorption, Distribution, Metabolism, and Excretion of Decursin and Decursinol Angelate from Angelica gigas Nakai

  • Kim, Kang-Min;Kim, Myo-Jeong;Kang, Jae-Seon
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1569-1572
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    • 2009
  • The pharmacokinetics of decursin and decursinol angelate (D/DA) were investigated in male SD rats following oral and intravenous administration. D/DA and metabolites obtained from in vitro samples were evaluated by LC/MS. The levels of D/DA and metabolized decursinol in the blood following oral and intravenous administrations declined according to first-order kinetics, with $T_{1/2}$ values of 56.67, 58.01, and 57.22 h, respectively, being observed after administration of a dose of 2 mg/kg body weight. The large intestine was the major site of disposition following oral administration. These data indicate that D/DA is rapidly absorbed from the gastrointestinal tract. In in vitro experiment utilizing liver microsomal protein, the major metabolic reaction of D/DA occurred to change decursinol. The cumulative biliary, urinary, and fecal excretions of D/DA in bile duct-cannulated rats was $36.10{\pm}2.9%$, $25.35{\pm}3.8%$, and $34.20{\pm}3.2%$, respectively, at 72 h after administration. These results indicate that the absorption of D/DA is almost complete, and that its metabolites are primarily excreted into feces through the bile. These results indicate that D/DA is subject to enterohepatic circulation.