• 제목/요약/키워드: metabolite analysis

검색결과 447건 처리시간 0.027초

Proteomic Dissection of Abiotic Stress Response in Crop Plants

  • Alam, Iftekhar;Sharmin, Shamima Akhtar;Lee, Byung-Hyun
    • 한국환경농학회:학술대회논문집
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    • 한국환경농학회 2011년도 30주년 정기총회 및 국제심포지엄
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    • pp.196-204
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    • 2011
  • Abiotic stress is the primary cause of crop loss worldwide, reducing average yields for most major crop plants by more than 50%. In addition, future agricultural production and management will encounter multifaceted challenges from global climate change. Therefore, it is necessary to study the molecular response of crop plants to the stresses in order to develop appropriate strategies to sustain food production under adverse environmental conditions. We carried out a large scale proteomic analysis of soybean plants in response to various abiotic stresses, including drought, salinity, waterlogging and their interactions. Proteins were analyzed by two dimensional polyacrylamide gel electrophoresis followed by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry. The identified proteins are involved in a wide range of cellular functions. In addition to the well known stress-associated proteins, we identified several novel proteins, which were not reported before. In many cases our proteomic data bridges the gap between mRNA and metabolite data. Our studie provides new insights into identification of abiotic stress responsive proteins in soybean, and demonstrates the advantages of proteomic analysis in dissecting metabolic and regulatory networks.

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Pharmacokinetic Changes in Drugs during Protein-Calorie Malnutrition: Correlation between Drug Metabolism and Hepatic Microsomal Cytochrome P450 Isozymes

  • Lee, Joo-Hyun;Suh, Ok-Kyung;Lee, Myung-Gull
    • Archives of Pharmacal Research
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    • 제27권7호
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    • pp.693-712
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    • 2004
  • The rats with protein-calorie malnutrition (PCM, 5% casein diet for a period of 4-week) were reported to exhibit 60 and 80% suppression in the hepatic microsomal cytochrome P450 (CYP) 1 A2 and CYP2C11 levels, respectively, and 40-50% decreases in CYP2E1 and CYP3A 1/2 levels compared to control (23% casein diet for a period of 4-week) based on Western blot analysis. In addition, Northern blot analysis showed that CYP1 A2, CYP2E1, CYP2C11, and CYP3A1/2 mRNAs decreased in the state of PCM as well. Hence, pharmacokinetic changes of the drugs in rats with PCM [especially the area under the plasma concentration-time curve from time zero to time infinity (AUC) changes of metabolite(s)] reported from literatures were tried to explain in terms of CYP isozyme changes in the rats. Otherwise, the time-averaged nonrenal clearance ($CL_{NR}$) of parent drug was compared. Pharmacokinetic changes of the drugs in other types of malnutritional state, such as kwashiorkor and marasmus, in both human and animal models were also compared. The drugs reviewed are as follows: diuretics, antibiotics, anticancer agents, antiepileptics, antiarrythmics, analgesics, xanthines, antimalarials, and miscellaneous.

Evaluation of PUN1 gene and capsaicinoids content in pepper genetic resources with excellent phenotype

  • Ro, Na-young;Hur, Onsook;Sung, Jungsook;Lee, Jeaeun;Hwang, Aejin;Lee, Hosun;Roh, Jaejong;Rhee, Juhee
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2019년도 춘계학술대회
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    • pp.69-69
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    • 2019
  • Pepper (Capsicum spp.) is one of the main quality features of this crop because of its sense of pungency, which is due to the presence of capsaicinoids. This compound is synthesized as a secondary metabolite and found only in the placental tissue of spicy fruit (Suzuki et al., 1980). Stewart et al. (2005) concluded that Pun1 encodes for the acyltransferase AT3 and they demonstrated its involvement in capsaicinoids metabolism. It was analyzed that the capsaicinoids content and PUN1 genotype in pepper genetic resources which were selected with excellent phenotype in field evaluation. The number of pepper genetic resources analyzed was 135, and species were C. annuum, C. baccatum, C. chinense, C. frutescens. The content of capsaicinoid ranged from 0 mg/100g to 828 mg/100g. The content of 0 mg/100g was the sweet pepper type, the highest content is IT 158530, the capsaicinoid content of which was 828 mg/100g and species was C. annuum. PUN1 gene analysis showed 117 pungent, 5 hetero, and 13 non-pungent. PUN1 analysis showed that 5 out of 13 non-pungent accessions were detected with low levels of capsaicinoid.

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A Genome-Wide Analysis of Antibiotic Producing Genes in Streptomyces globisporus SP6C4

  • Kim, Da-Ran;Kwak, Youn-Sig
    • The Plant Pathology Journal
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    • 제37권4호
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    • pp.389-395
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    • 2021
  • Soil is the major source of plant-associated microbes. Several fungal and bacterial species live within plant tissues. Actinomycetes are well known for producing a variety of antibiotics, and they contribute to improving plant health. In our previous report, Streptomyces globisporus SP6C4 colonized plant tissues and was able to move to other tissues from the initially colonized ones. This strain has excellent antifungal and antibacterial activities and provides a suppressive effect upon various plant diseases. Here, we report the genome-wide analysis of antibiotic producing genes in S. globisporus SP6C4. A total of 15 secondary metabolite biosynthetic gene clusters were predicted using antiSMASH. We used the CRISPR/Cas9 mutagenesis system, and each biosynthetic gene was predicted via protein basic local alignment search tool (BLAST) and rapid annotation using subsystems technology (RAST) server. Three gene clusters were shown to exhibit antifungal or antibacterial activity, viz. cluster 16 (lasso peptide), cluster 17 (thiopeptide-lantipeptide), and cluster 20 (lantipeptide). The results of the current study showed that SP6C4 has a variety of antimicrobial activities, and this strain is beneficial in agriculture.

Metabolomics reveals potential plateau adaptability by regulating inflammatory response and oxidative stress-related metabolism and energy metabolism pathways in yak

  • Huang, Meizhou;Zhang, Xin;Yan, Wenjun;Liu, Jingjing;Wang, Hui
    • Journal of Animal Science and Technology
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    • 제64권1호
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    • pp.97-109
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    • 2022
  • Species are facing strong selection pressures to adapt to inhospitable high-altitude environments. Yaks are a valuable species and an iconic symbol of the Qinghai-Tibet Plateau. Extensive studies of high-altitude adaptation have been conducted, but few have focused on metabolism. In the present study, we determined the differences in the serum metabolomics between yaks and the closely related species of low-altitude yellow cattle and dairy cows. We generated high-quality metabolite profiling data for 36 samples derived from the three species, and a clear separation trend was obtained between yaks and the other animals from principal component analysis. In addition, we identified a total of 63 differentially expressed metabolites among the three species. Functional analysis revealed that differentially expressed metabolites were related to the innate immune activation, oxidative stress-related metabolism, and energy metabolism in yaks, which indicates the important roles of metabolites in high-altitude adaptation in yaks. The results provide new insights into the mechanism of adaptation or acclimatization to high-altitude environments in yaks and hypoxia-related diseases in humans.

Screening of the liver, serum, and urine of piglets fed zearalenone using a NMR-based metabolomic approach

  • Jeong, Jin Young;Kim, Min Seok;Jung, Hyun Jung;Kim, Min Ji;Lee, Hyun Jeong;Lee, Sung Dae
    • 농업과학연구
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    • 제45권3호
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    • pp.447-454
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    • 2018
  • Zearalenone (ZEN), a mycotoxin produced by Fusarium in food and feed, causes serious damage to the health of humans and livestock. Therefore, we compared the metabolomic profiles in the liver, serum, and urine of piglets fed a ZEN-contaminated diet using proton nuclear magnetic resonance ($^1H-NMR$) spectroscopy. The spectra from the three different samples, treated with ZEN concentrations of 0.8 mg/kg for 4 weeks, were aligned and identified using MATLAB. The aligned data were subjected to discriminating analysis using multivariate statistical analysis and a web server for metabolite set enrichment analysis. The ZEN-exposed groups were almost separated in the three different samples. Metabolic analysis showed that 28, 29, and 20 metabolites were profiled in the liver, serum, and urine, respectively. The discriminating analysis showed that the alanine, arginine, choline, and glucose concentrations were increased in the liver. Phenylalanine and tyrosine metabolites showed high concentrations in serum, whereas valine showed a low concentration. In addition, the formate levels were increased in the ZEN-treated urine. For the integrated analysis, glucose, lactate, taurine, glycine, alanine, glutamate, glutamine, and creatine from orthogonal partial least squares discriminant analysis (OPLS-DA) were potential compounds for the discriminating analysis. In conclusion, our findings suggest that potential biomarker compounds can provide a better understanding on how ZEN contaminated feed in swine affects the liver, serum, and urine.

GC/MS를 이용한 소변 중 대마 대사체 분석의 측정불확도 평가 (Uncertainty Evaluation of the Analysis of 11-Nor-9-carboxy-${\Delta}^9$-tetrahydrocannabinol in Human Urine by GC/MS)

  • 김진영;정재철;서승일;서용준;이정직;김종상;인문교
    • 약학회지
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    • 제52권6호
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    • pp.480-487
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    • 2008
  • We described an estimation of measurement uncertainty in quantitative analysis of 11-nor-9-carboxy-${\Delta}^9$-tetrahydrocannabinol (THCCOOH), the major metabolite of ${\Delta}^9$-tetrahydrocannabinol, in urine sample by solid-phase extraction (SPE) and GC/MS detection. The analytical results were compared and the different contributions to the uncertainty were evaluated. Inter-day and inter-person validation were performed using statistical analysis of several indicative factors. Measurement uncertainty associated with target analyte in real forensic samples was estimated using quality control (QC) data. Traceability of measurement was established through traceable standards, calibrated volumetric glassware and volume measuring device. The major factors of contribution to combined standard uncertainty, were calibration linearity, inter-day repeatability and inter-person reproducibility, while those associated with preparation of analytical standards and sampling volume were not so important considering the degree of contribution. Relative combined standard uncertainties associated with the described method was 12.05% for THCCOOH.

Tentative identification of 20(S)-protopanaxadiol metabolites in human plasma and urine using ultra-performance liquid chromatography coupled with triple quadrupole time-of-flight mass spectrometry

  • Ling, Jin;Yu, Yingjia;Long, Jiakun;Li, Yan;Jiang, Jiebing;Wang, Liping;Xu, Changjiang;Duan, Gengli
    • Journal of Ginseng Research
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    • 제43권4호
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    • pp.539-549
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    • 2019
  • Background: 20(S)-Protopanaxadiol (PPD), the aglycone part of 20(S)-protopanaxadiol ginsenosides, possesses antidepressant activity among many other pharmacological activities. It is currently undergoing clinical trial in China as an antidepressant. Methods: In this study, an ultra-performance liquid chromatography coupled with triple quadrupole time-of-flight mass tandem mass spectrometry method was established to identify the metabolites of PPD in human plasma and urine following oral administration in phase IIa clinical trial. Results: A total of 40 metabolites in human plasma and urine were identified using this method. Four metabolites identified were isolated from rat feces, and two of them were analyzed by NMR to elucidate the exact structures. The structures of isolated compounds were confirmed as (20S,24S)-epoxydammarane-12,23,25-triol-3-one and (20S,24S)-epoxydammarane-3,12,23,25-tetrol. Both compounds were found as metabolites in human for the first time. Upon comparing our findings with the findings of the in vitro study of PPD metabolism in human liver microsomes and human hepatocytes, metabolites with m/z 475.3783 and phase II metabolites were not found in our study whereas metabolites with m/z 505.3530, 523.3641, and 525.3788 were exclusively detected in our experiments. Conclusion: The metabolites identified using ultra-performance liquid chromatography coupled with triple quadrupole time-of-flight mass spectrometry in our study were mostly hydroxylated metabolites. This indicated that PPD was metabolized in human body mainly through phase I hepatic metabolism. The main metabolites are in 20,24-oxide form with multiple hydroxylation sites. Finally, the metabolic pathways of PPD in vivo (human) were proposed based on structural analysis.

LC/TOFMS를 이용한 생체시료의 내인성 대사체 분석법 개발 (Method Development for the Profiling Analysis of Endogenous Metabolites by Accurate-Mass Quadrupole Time-of-Flight(Q-TOF) LC/MS)

  • 이인선;김진호;조수열;심선보;박혜진;이진희;이지현;황인선;김성일;이정희;조수연;최돈웅;조양하
    • 한국식품위생안전성학회지
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    • 제25권4호
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    • pp.388-394
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    • 2010
  • Metabolomics aims at the comprehensive, qualitative and quantitative analysis of wide arrays of endogenous metabolites in biological samples. It has shown particular promise in the area of toxicology and drug development, functional genomics, system biology and clinical diagnosis. In this study, analytical technique of MS instrument with high resolution mass measurement, such as time-of-flight (TOF) was validated for the purpose of investigation of amino acids, sugars and fatty acids. Rat urine and serum samples were extracted by selected each solvent (50% acetonitrile, 100% acetonitrile, acetone, methanol, water, ether) extraction method. We determined the optimized liquid chromatography/time-of-flight mass spectrometry (LC/TOFMS) system and selected appropriated columns, mobile phases, fragment energy and collision energy, which could search 17 metabolites. The spectral data collected from LC/TOFMS were tested by ANOVA. Obtained with the use of LC/TOFMS technique, our results indicated that (1) MS and MS/MS parameters were optimized and most abundant product ion of each metabolite were selected to be monitorized; (2) with design of experiment analysis, methanol yielded the optimal extraction efficiency. Therefore, the results of this study are expected to be useful in the endogenous metabolite fields according to validated SOP for endogenous amino acids, sugars and fatty acids.

타액 중 ${\Delta}^9$-Tetrahydrocannabinol 및 11-Nor-9-carboxy-${\Delta}^9$-Tetrahydrocannabinol의 분석법 확립 및 안정성 검토 (Development of Quantification Method and Stability of ${\Delta}^9$-Tetrahydrocannabinol and 11-Nor-9-carboxy-${\Delta}^9$-Tetrahydrocannabinol in Oral Fluid)

  • 최혜영;백승경;장문희;최화경;정희선
    • 약학회지
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    • 제54권4호
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    • pp.226-231
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    • 2010
  • Oral fluid has become increasingly popular as an alternative specimen in the field of driving under the influence of drugs (DUID) and work place drug testing. In this study, an analytical method for the detection and quantification of ${\Delta}^9$-tetrahydrocannabinol (THC) and its metabolite, 11-nor-9-carboxy-${\Delta}^9$-tetrahydrocannabinol (THC-COOH) in oral fluid by SPE and GC-MS was established and fully validated. The stability of THC and THC-COOH in oral fluid during storage was also determined by examining the THC and THC-COOH concentration changes depending on time and container materials. Oral fluid samples were kept over 21 days at room temperature, $-4^{\circ}C$ and $-20^{\circ}C$ in two different specimen collection tubes; glass and polypropylene tubes. Three replicates for each condition with different temperature and types of a container were analyzed at five different time points over 21 days. When oral fluid samples were stored in glass tubes, the loss of both THC and THC-COOH was less than 10% at all room temperature, $-4^{\circ}C$ and $-20^{\circ}C$. However, in polypropylene tubes, the loss of both THC and THC-COOH increased significantly over the study period. In particular, the concentration of THC decreased more rapidly than that of THC-COOH at room temperature and the maximal percentage of THC lost was 90.3% after 21 days. The result indicates that it would be necessary to collect oral fluid samples in glass containers and cool the samples until analysis in order to prevent the degradation of analytes.