• Title/Summary/Keyword: metabolite analysis

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Analysis of metabolites in wheat roots in response to salinity stress

  • Kim, Da-Eun;Roy, Swapan Kumar;Kim, Ki-Hyun;Cho, Seong-Woo;Park, Chul-Soo;Lee, Moon-Soon;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.200-200
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    • 2017
  • Salinity stress is one of the most important abiotic stresses and severely impairs plant growth and production. Root is the first site for nutrient accumulation like as $Na^+$ in the plant. To investigate the response of wheat root under salinity stress, we executed the characterization of morphology and analysis of metabolites. Wheat seeds cv. Keumgang (Korean cultivar) were grown on the moist filter paper in Petri dish. After 5 days, seedlings were transferred to hydroponic apparatus at 1500 LUX light intensity, at $20^{\circ}C$ with 70% relative humidity in a growth chamber. Seedlings (5-day-old) were exposed to 50mM, 75mM, 100mM NaCl for 5 days. Ten-day-old seedlings were used for morphological characterization and metabolite analysis. Root and leaf length became shorter in high NaCl concentration compared to following NaCl treatment. For confirmation of salt accumulation, wheat roots were stained with $CoroNa^+$ Green AM, and fluoresce, and the image was taken by confocal microscopy. $Na^+$ ion accumulation rate was higher at 100mM compared to the untreated sample. Furthermore, to analyze metabolites in the wheat root, samples were extracted by $D_2O$ solvent, and extracted sample was analyzed by 1H NMR spectroscopy. Fourteen metabolites were identified in wheat roots using NMR spectroscopy. Methanol and ethanol were up-regulated, whereas formate, aspartate, aminobutyrate, acetate and valine were down-regulated under salinity stress on roots of wheat. Fumarate had no change, while glucose, betaine, choline, glutamate and lactate were unevenly affected during salinity stress.

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Metabolomic Analysis of Ethyl Acetate and Methanol Extracts of Blueberry (Ethyl Acetate와 Methanol을 이용한 블루베리 추출물 대사체 분석)

  • Jo, Young-Hee;Kim, Sugyeong;Kwon, Da-Ae;Lee, Hong Jin;Choi, Hyung-Kyoon;Auh, Joong-Hyuck
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.419-424
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    • 2014
  • Metabolite profiling of blueberry (cultivar "Spartan") was performed by extraction using different solvents, methanol and ethyl acetate, through metabolomic analysis using LC-MS/MS. Unsupervised classification method (PCA) and supervised prediction model (OPLS-DA) provided good categorization of metabolites according to the extraction solvents. Metabolites of the anthocyanin family, including delphinidin hexoside, delphinidin, 5-O-feruloylquinic acid, malvidin hexoside, malvidin-3-arabinoside, petunidin-3-arabinoside, and petunidin hexoside, were mainly detected in methanol fractions, whereas those of the flavonoid family, including chlorogenic acid, chlorogenic acid dimer, 6,8-di-C-arabinopyranosyl-luteolin, and luteolin were successfully prepared in the ethyl acetate fraction. Thus, metabolomic analysis of blueberry extracts allows for the simple profiling of whole and distinctive metabolites for future applications.

Co-cultured methanogen improved the metabolism in the hydrogenosome of anaerobic fungus as revealed by gas chromatography-mass spectrometry analysis

  • Li, Yuqi;Sun, Meizhou;Li, Yuanfei;Cheng, Yanfen;Zhu, Weiyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.12
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    • pp.1948-1956
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    • 2020
  • Objective: The purpose of this study was to reveal the metabolic shift in the fungus cocultured with the methanogen (Methanobrevibacter thaueri). Methods: Gas chromatography-mass spectrometry was used to investigate the metabolites in anaerobic fungal (Pecoramyces sp. F1) cells and the supernatant. Results: A total of 104 and 102 metabolites were detected in the fungal cells and the supernatant, respectively. The partial least squares-discriminant analysis showed that the metabolite profiles in both the fungal cell and the supernatant were distinctly shifted when co-cultured with methanogen. Statistically, 16 and 30 metabolites were significantly (p<0.05) affected in the fungal cell and the supernatant, respectively by the co-cultured methanogen. Metabolic pathway analysis showed that co-culturing with methanogen reduced the production of lactate from pyruvate in the cytosol and increased metabolism in the hydrogenosomes of the anaerobic fungus. Citrate was accumulated in the cytosol of the fungus co-cultured with the methanogen. Conclusion: The co-culture of the anaerobic fungus and the methanogen is a good model for studying the microbial interaction between H2-producing and H2-utilizing microorganisms. However, metabolism in hydrogenosome needs to be further studied to gain better insight in the hydrogen transfer among microorganisms.

Mass Cultivation and Secondary Metabolite Analysis of Rhodobacter capsulatus PS-2 (광합성세균 Rhodobater capsulatus PS-2의 대량배양 최적화 및 대사산물 분석)

  • Bong, Ki Moon;Kim, Jong Min;Yoo, Jae-Hong;Park, In Chul;Lee, Chul Won;Kim, Pyoung Il
    • KSBB Journal
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    • v.31 no.3
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    • pp.158-164
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    • 2016
  • Plant growth promoting (PGP) hormones, which are produced in a small quantity by bacteria, affect in plant growth and development. PGPs play an important role on the crop productivity in agricultural field. In this study, a photosynthetic bacterial strain producing the PGP was isolated from paddy soil. Bacterial isolate was gram negative, rod-shaped and motility positive. From the 16s rRNA gene sequence analysis, the isolate was identified as Rhodobacter capsulatus PS-2. The mass cultivation of R. capsulatus PS-2 was optimized by considering of the carbon, nitrogen and inorganic salt sources. Optimal medium composition was determined as Na-succinate 4.5 g, yeast extract 5 g, $K_2HPO_4$ 1 g, $MgSO_4$ 5 g, per liter. From the result of 500 L fermentation for 2 days using the optimal medium, the viable cells were $8.7{\times}10^9cfu/mL$. R. capsulatus PS-2 strain produced the carotenoid and indole-3-acetic acid (IAA). The carotenoid extraction and quantitative analysis were performed by HCl-assisting method. Total carotenoid contents from R. capsulatus PS-2 culture broth were measured as $7.02{\pm}0.04$ and $6.93{\pm}0.05mg/L$ under photoheterotrophic and chemoheterotrophic conditions, respectively. To measure the productivity of IAA, colorimetric method was employed using Salkowski reagent at optical density 535 nm. The results showed that the highest content of IAA was $197.44{\pm}5.92mg/L$ in the optimal medium supplemented with 0.3% tryptophan.

Isolation and Characterization of Comprehensive Polychlorinated Biphenyl-Degrading Bacterium, Enterobacter sp. LY402

  • Jia, Ling-Yun;Zheng, Ai-Ping;Xu, Li;Huang, Xiao-Dong;Zhang, Qing;Yang, Feng-Lin
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.952-957
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    • 2008
  • A Gram-negative bacterium, named LY402, was isolated from contaminated soil. 16S rDNA sequencing and measurement of the physiological and biochemical characteristics identified it as belonging to the genus Enterohacter. Degradation experiments showed that LY402 had the ability to aerobically transform 79 of the 91 major congeners of Aroclor 1242, 1254, and 1260. However, more interestingly, the strain readily degraded certain highly chlorinated and recalcitrant polychlorinated biphenyls (PCBs). Almost all the tri- and tetra-chlorobiphenyls (CBs), except for 3,4,3',4'-CB, were degraded in 3 days, whereas 73% of 3,4,3',4'-, 92% of the penta-, 76% of the hexa-, and 37% of the hepta-CBs were transformed after 6 days. In addition, among 12 octa-CBs, 2,2',3,3',5,5',6,6-CB was obviously degraded, and 2,2',3,3',4,5,6,6'- and 2,2',3,3',4,5,5',6'-CB were slightly transformed. In a metabolite analysis, mono- and dichlorobenzoic acids (CBAs) were identified, and parts of them were also transformed by strain LY402. Analysis of PCB degradation indicated that strain LY402 could effectively degrade PCB congeners with chlorine substitutions in both ortho- and para-positions. Consequently, this is the first report of an Enterobacteria that can efficiently degrade both low and highly chlorinated PCBs under aerobic conditions.

Effect of Combined Exposure to EDTA and Zinc Pyrithione on Pyrithione Absorption in Rats

  • Jung, Dong Sik;Jung, Guk Hwa;Lee, Eun Ho;Park, Hye Ran;Kim, Ju Hwan;Kim, Kyu-Bong;Kim, Hak Rim;Kim, Hyung Gun
    • Toxicological Research
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    • v.35 no.2
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    • pp.155-160
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    • 2019
  • Zinc pyrithione (ZnPT) is a coordination complex of zinc and has been used widely as an anti-dandruff agent in shampoos. Many shampoos contain both ZnPT and EDTA, a chelating agent speculated to increase ZnPT absorption, thereby raising concerns about neurotoxicity. Here, we investigated the effect of EDTA on ZnPT absorption by direct comparison of ZnPT and pyrithione (PT) concentrations in shampoo formulations, and by pharmacokinetic analysis of ZnPT, PT, and 2-methanesulfonylpyridine (MSP), the main ZnPT metabolite, in rat plasma or urine following exposure to shampoo containing ZnPT alone or a combination of ZnPT and EDTA. Approximately 17.3% of ZnPT was converted to PT by the addition of EDTA in the shampoo formulation. Plasma ZnPT and PT concentrations were not measured up to 24 hr after treatment with shampoo containing 1% ZnPT or 1% ZnPT + 2% EDTA in all rats. However, PT amount in 24-hr urine sample, MSP concentration in plasma, and MSP amount in 24-hr urine sample were approximately 4-, 2.6-, and 2.7-fold higher, respectively, in the 1% ZnPT + 2% EDTA shampoo group than in the 1% ZnPT shampoo group. As confirmed by the formulation analysis and in vivo pharmacokinetic analysis, the exposure of ZnPT could be increased by the absorption of PT due to partial dissociation of ZnPT into PT.

Metabolomics Approach to Explore the Effects of Rebamipide on Inflammatory Arthritis Using Ultra Performance Liquid Chromatography/Quadrupole Time-of-Flight Mass Spectrometry

  • Moon, Su-Jin;Lee, Soo Hyun;Jung, Byung-Hwa;Min, Jun-Ki
    • Journal of Rheumatic Diseases
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    • v.24 no.4
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    • pp.192-202
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    • 2017
  • Objective. Rebampide is a gastroprotective agent used to treat gastritis. It possesses anti-inflammatory and anti-arthritis effects, but the mechanisms of these effects are not well understood. The objective of this study was to explore mechanisms underlying the therapeutic effects of rebamipide in inflammatory arthritis. Methods. Collagen-induced arthritis (CIA) was induced in DBA/1J mice. DBA/1J mice were immunized with chicken type II collagen, then treated intraperitoneally with rebamipide (10 mg/kg or 30 mg/kg) or vehicle (10% carboxymethylcellulose solution) alone. Seven weeks later, plasma samples were collected. Plasma metabolic profiles were analyzed using ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry-based metabolomics study and metabolite biomarkers were identified through multivariate data analysis. Results. Low dose rebamipide treatment reduced the clinical arthritis score compared with vehicle treatment, whereas high dose rebamipide in CIA aggravated arthritis severity. Based on multivariate analysis, 17 metabolites were identified. The plasma levels of metabolites associated with fatty acids and phospholipid metabolism were significantly lower with rebamipide treatment than with vehicle. The levels of $15-deoxy-^{{\Delta}12,14}$ prostaglandin J2 and thromboxane B3 decreased only in high dose-treated groups. Certain peptide molecules, including enterostatin (VPDPR) enterostatin and bradykinin dramatically increased in rebamipide-treated groups at both doses. Additionally, corticosterone increased in the low dose-treated group and decreased in the high dose-treated group. Conclusion. Metabolomics analysis revealed the anti-inflammatory effects of rebamipide and suggested the potential of the drug repositioning in metabolism- and lipid-associated diseases.

Liquid Chromatography-Tandem Mass Spectrometric Analysis of Nannozinone A and Its Application to Pharmacokinetic Study in Mice

  • Lee, Chul Haeng;Kim, Soobin;Lee, Jaehyeok;Jeon, Ji-Hyeon;Song, Im-Sook;Han, Young Taek;Choi, Min-Koo
    • Mass Spectrometry Letters
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    • v.12 no.1
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    • pp.21-25
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    • 2021
  • We aimed to develop and validate a sensitive analytical method of nannozinone A, active metabolite of Nannochelins A extracted from the Myxobacterium Nannocytis pusilla, in mouse plasma using a liquid chromatography-tandem mass spectrometry (LC-MS/MS). Mouse plasma samples containing nannozinone A and 13C-caffeine (internal standard) were extracted using a liquid-liquid extraction (LLE) method with methyl tert-butyl ether. Standard calibration curves were linear in the concentration range of 1 - 1000 ng/mL (r2 > 0.998) with the inter- and intra-day accuracy and precision results less than 15%. LLE method gave results in the high and reproducible extraction recovery in the range of 78.00-81.08% with limited matrix effect in the range of 70.56-96.49%. The pharmacokinetics of nannozinone A after intravenous injection (5 mg/kg) and oral administration (30 mg/kg) of nannozinone A were investigated using the validated LC-MS/MS analysis of nannozinone A. The absolute oral bioavailability of nannozinone A was 8.82%. Plasma concentration of nannozinone A after the intravenous injection sharply decreased for 4 h but plasma concentration of orally administered nannozinone A showed fast distribution and slow elimination for 24 h. In conclusion, we successfully applied this newly developed sensitive LC-MS/MS analytical method of nannozinone A to the pharmacokinetic evaluation of this compound. This method can be useful for further studies on the pharmacokinetic optimization and evaluating the druggability of nannozinone A including its efficacy and toxicity.

Allometric analysis of tylosin tartrate pharmacokinetics in growing male turkeys

  • Pozniak, Blazej;Tikhomirov, Marta;Motykiewicz-Pers, Karolina;Bobrek, Kamila;Switala, Marcin
    • Journal of Veterinary Science
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    • v.21 no.3
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    • pp.35.1-35.11
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    • 2020
  • Background: Despite common use of tylosin in turkeys, the pharmacokinetic (PK) data for this drug in turkeys is limited. Within a few months of growth, PK of drugs in turkeys undergoes changes that may decrease their efficacy due to variable internal exposure. Objectives: The objective of this study was to investigate the influence of age on the PK of a single intravenous (i.v.) and oral administration of tylosin to turkeys at a dose of 10 and 50 mg/kg, respectively. Methods: Plasma drug concentrations were measured using high-performance liquid chromatography with UV detection. The PK parameters were assessed by means of non-compartmental approach and were subjected to allometric analysis. Results: During a 2.5-month-long period of growth from 1.4 to 14.7 kg, the median value for area under the concentration-time curve after i.v. administration increased from 2.61 to 7.15 mg × h/L and the body clearance decreased from a median of 3.81 to 1.42 L/h/kg. Over the same time, the median elimination half-life increased from 1.03 to 2.96 h. For the oral administration a similar trend was noted but the differences were less pronounced. Bioavailability was variable (5.76%-21.59%) and age-independent. For both routes, the plasma concentration of the major tylosin metabolite, tylosin D, was minimal. Protein binding was age-independent and did not exceed 50%. Allometric analysis indicated a relatively poor predictivity of clearance, volume of distribution and elimination half-life for tylosin in turkeys. Conclusions: Age has a significant impact on tylosin PK in turkeys and dosage adjustment may be needed, particularly in young individuals.

The Evaluation Functional Activity and Indicator Component Analysis and According to the Extraction Method of Eleuthrococcus Gracilistylus (섬오갈피 추출물 중 acanthoic acid 및 Kaurenoic acid 함량 분석 및 기능성 활성 평가)

  • Hyun Kyoung Kim
    • The Journal of the Convergence on Culture Technology
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    • v.10 no.2
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    • pp.73-80
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    • 2024
  • To determinate the content of acanthoic acid and kaurenoic acid in 70% EtOH and hot water extracts of Eleutherococcus gracilistylus, quantitative analysis of each compound in samples was carried out by a HPLC-UVD. Also, the identification of each acompound in samples was successfully assigned by LC-MS analysis. In result, the contents of acanthoic acid and kaurenoic acid in 70% ethanoic extracts were 28.84±0.21 mg/g (2.88%), 26.38±1.63 mg/g (2.64%), respectively. However, the content of two compounds in hot-water extracts was not observed. In conclusion, it shows that 70% ethanol as a best extraction solvent to extract the acanthoic acid and its metabolite from Eleutherococcus gracilistylus was better than hot-water solvent. The 70% ethanol complex extract of Allium Hookeri and Eleutherococcus gracilistylus showed better effectiveness. In addition, the 70% ethanol extract complex of Allium Hookeri and Eleutherococcus gracilistylus showed better effects than the hot water solvent of DPPH radical scavenging ability, total polyphenols, and flavonoids content. The anti-inflammatory activity were significantly or partially reduced by treatment with ethanol extract complex(SEC) by Allium Hookeri and Eleutherococcus gracilistylus.