• Title/Summary/Keyword: Functional metabolite

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Comparative metabolomic analysis in horses and functional analysis of branched chain (alpha) keto acid dehydrogenase complex in equine myoblasts under exercise stress

  • Jeong-Woong, Park;Kyoung Hwan, Kim;Sujung, Kim;Jae-rung, So;Byung-Wook, Cho;Ki-Duk, Song
    • Journal of Animal Science and Technology
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    • v.64 no.4
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    • pp.800-811
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    • 2022
  • The integration of metabolomics and transcriptomics may elucidate the correlation between the genotypic and phenotypic patterns in organisms. In equine physiology, various metabolite levels vary during exercise, which may be correlated with a modified gene expression pattern of related genes. Integrated metabolomic and transcriptomic studies in horses have not been conducted to date. The objective of this study was to detect the effect of moderate exercise on the metabolomic and transcriptomic levels in horses. In this study, using nuclear magnetic resonance (NMR) spectroscopy, we analyzed the concentrations of metabolites in muscle and plasma; we also determined the gene expression patterns of branched chain (alpha) keto acid dehydrogenase kinase complex (BCKDK), which encodes the key regulatory enzymes in branched-chain amino acid (BCAA) catabolism, in two breeds of horses, Thoroughbred and Jeju, at different time intervals. The concentrations of metabolites in muscle and plasma were measured by 1H NMR (nuclear magnetic resonance) spectroscopy, and the relative metabolite levels before and after exercise in the two samples were compared. Subsequently, multivariate data analysis based on the metabolic profiles was performed using orthogonal partial least square discriminant analysis (OPLS-DA), and variable important plots and t-test were used for basic statistical analysis. The stress-induced expression patterns of BCKDK genes in horse muscle-derived cells were examined using quantitative reverse transcription polymerase chain reaction (qPCR) to gain insight into the role of transcript in response to exercise stress. In this study, we found higher concentrations of aspartate, leucine, isoleucine, and lysine in the skeletal muscle of Jeju horses than in Thoroughbred horses. In plasma, compared with Jeju horses, Thoroughbred horses had higher levels of alanine and methionine before exercise; whereas post-exercise, lysine levels were increased. Gene expression analysis revealed a decreased expression level of BCKDK in the post-exercise period in Thoroughbred horses.

Varying Effects of Artificial Light on Plant Functional Metabolites (인공광 이용에 따른 작물의 기능성 물질의 차별적 증가)

  • Kim, Yang Min;Sung, Jwa Kyung;Lee, Ye Jin;Lee, Deog Bae;Yoo, Chul Hyun;Lee, Seul Bi
    • Korean Journal of Environmental Agriculture
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    • v.38 no.1
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    • pp.61-67
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    • 2019
  • BACKGROUND: Many studies on artificial lighting have been recently performed to investigate its effect on agricultural products with good quality. This review was aimed at comparing the effects of artificial light on functional metabolites of the plants that were grown in greenhouses and growth chamber. METHODS AND RESULTS: It has been summarized that artificial lighting both in growth chambers and greenhouses caused different functional metabolites patterns depending on light quality. Even though the same light quality was applied, different patterns in metabolites were observed in different plant species. For the same species, supplementation of the same light quality in both growth chambers and greenhouses did cause different functional metabolites patterns. CONCLUSION: Artificial lighting caused different patterns in functional metabolites of plants grown in greenhouses and growth chambers, depending on the light quality and/or plant species. The manipulation of plant growth and functional metabolites would be possible by engineering the light qualities, but knowledge on proper lighting condition depending on plant species and growth places would be necessary.

5β-Hydroxypalisadin B isolated from red alga Laurencia snackeyi attenuates inflammatory response in lipopolysaccharide-stimulated RAW 264.7 macrophages

  • Wijesinghe, W.A.J.P.;Kang, Min-Cheol;Lee, Won-Woo;Lee, Hyi-Seung;Kamada, Takashi;Vairappan, Charles S.;Jeon, You-Jin
    • ALGAE
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    • v.29 no.4
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    • pp.333-341
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    • 2014
  • In this study, four compounds isolated from the red alga Laurencia snackeyi were evaluated for their potential anti-inflammatory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. These compounds were tested for their inhibitory effects on nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells. Since $5{\beta}$-hydroxypalisadin B showed the best activity it was further tested for the production of prostaglandin-$E_2$ ($PGE_2$), expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), the release of pro-inflammatory cytokines tumor necrotic factor-alpha (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and interleukin-6 (IL-6). $5{\beta}$-Hydroxypalisadin B significantly reduced the $PGE_2$ release and suppressed the iNOS and COX-2 expression in LPS-stimulated RAW 264.7 cells. It also significantly reduced the release of pro-inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. These findings provide the first evidence of anti-inflammatory potential of $5{\beta}$-hydroxypalisadin B isolated from the red alga L. snackeyi and hence, it could be exploited as an active ingredient in pharmaceutical, nutraceutical and functional food applications.

The anti-tumor efficacy of 20(S)-protopanaxadiol, an active metabolite of ginseng, according to fasting on hepatocellular carcinoma

  • Li, Wenzhen;Wang, Yifan;Zhou, Xinbo;Pan, Xiaohong;Lu, Junhong;Sun, Hongliu;Xie, Zeping;Chen, Shayan;Gao, Xue
    • Journal of Ginseng Research
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    • v.46 no.1
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    • pp.167-174
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    • 2022
  • Background: 20(S)-protopanaxadiol (20(S)-PPD), one of the main active metabolites of ginseng, performs a broad spectrum of anti-tumor effects. Our aims are to search out new strategies to enhance anti-tumor effects of natural products, including 20(S)-PPD. In recent years, fasting has been shown to be multi-functional on tumor progression. Here, the effects of fasting combined with 20(S)-PPD on hepatocellular carcinoma growth, apoptosis, migration, invasion and cell cycle were explored. Methods: CCK-8 assay, trypan blue dye exclusion test, imagings photographed by HoloMonitorTM M4, transwell assay and flow cytometry assay were performed for functional analyses on cell proliferation, morphology, migration, invasion, apoptosis, necrosis and cell cycle. The expressions of genes on protein levels were tested by western blot. Tumor-bearing mice were used to evaluate the effects of intermittent fasting combined with 20(S)-PPD. Results: We firstly confirmed that fasting-mimicking increased the anti-proliferation effect of 20(S)-PPD in human HepG2 cells in vitro. In fasting-mimicking culturing medium, the apoptosis and necrosis induced by 20(S)-PPD increased and more cells were arrested at G0-G1 phase. Meanwhile, invasion and migration of cells were decreased by down-regulating the expressions of matrix metalloproteinase (MMP)-2 and MMP-9 in fasting-mimicking medium. Furthermore, the in vivo study confirmed that intermittent fasting enhanced the tumor growth inhibition of 20(S)-PPD in H22 tumor-bearing mice without obvious side effects. Conclusion: Fasting significantly sensitized HCC cells to 20(S)-PPD in vivo and in vitro. These data indicated that dietary restriction can be one of the potential strategies of chinese medicine or its active metabolites against hepatocellular carcinoma.

HPLC-UVD method validation for quantitative analysis of camelliaside A in hot-water extract of soybean (Glycine max L.) leaves (콩잎 열수추출물의 지표성분인 camelliaside A의 정량분석을 위한 HPLC-UVD 분석법 밸리데이션)

  • Kim, Jeong Ho;Lee, Seung Hwan;Moon, Si Won;Park, Ki Hun
    • Journal of Applied Biological Chemistry
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    • v.65 no.3
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    • pp.195-202
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    • 2022
  • Soybean (Glycine max L.) leaves have been researched as functional food stuff actively, but there is no validation method to control quality of soybean leaves (SL). In this study, we annotated seven kaempferol derivatives to confirm camelliaside A as index metabolite in SL using UHPLC-ESI-TOF-MS. HPLC-UVD validation method of camelliaside A in hot-water extract of SL was established according to validation guideline of functional foods from the Ministry of Food and Safety of Korea. The HPLC-UVD method was validated with reliable parameters for examining specificity, accuracy, precision, limit of detection and quantification and linearity. The established method gave the suitable ranges to qunatitate camelliaside A from the hot-water extract of soybean leaves.

Feasibility of Proton Chemical Shift Imaging with a Stereotactic Headframe

  • 백현만;최보영;손병철;정성택;이형구;서태석
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2003.09a
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    • pp.72-72
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    • 2003
  • Purpose: To prove feasibility of proton chemical shift imaging (lH CSI) during stereotactic procedure, authors performed IH CSI in combination with a stereotactic headframe and selected targets according to local metabolic information, evaluated the pathologic results. Methods: The 1H CSI directed stereotactic biopsy was performed in five patients. 1H CSI was performed before conventional stereotactic MRI with gadolinium enhancement for stereotactic coordinates. The metabolite images expressed as integral ratios, Cho/Cr and Lac/Cr, were displayed in different colors. The stereotactic target coordinates were correlated with the coordinates from the 1H CSI images. Results: The final pathologic results obtained were concordant with the local metabolic information from 1H CSI. We believe that 1H CSI-directed stereotatic biopsy has the potential to significantly improve the accuracy of stereotactic biopsy targeting. Conclusions : Metabolic signals derived from 1H CSI could give us more direct clues for stereotactic target selection during the subsequent conventional stereotactic MR imaging. 1H CSI was feasible with the stereotatic headframe in place. The final pathologic results obtained were concordant with the local metabolic information from 1H CSI. Acknowledgement: This study was supported by a grant of the Center for Functional and Metabolic Imaging Technology, Ministry of Health & Welfare, Republic of Korea (02-PJ3-PG6-EV07-0002).

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Estrogenic Compounds Compatible with a Conditional Gene Expression System for the Phytopathogenic Fungus Fusarium graminearum

  • Lee, Jung-Kwan;Son, Ho-Kyoung;Lee, Yin-Won
    • The Plant Pathology Journal
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    • v.27 no.4
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    • pp.349-353
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    • 2011
  • The ascomycete fungus Fusarium graminearum is an important plant pathogen responsible for Fusarium head blight in small grains and ear rot on maize. This fungus also produces the estrogenic metabolite, zearalenone (ZEA) that causes estrogenic disorders in humans and animals. Previously, we developed a conditional gene expression system for this fungus using a ZEA-inducible promoter (Pzear). In the present study, four other estrogenic compounds, including ${\beta}$-estradiol, estriol, estrone, and secoisolariciresinol, were screened as possible substitutes for ZEA in this system. Among them, ${\beta}$-estradiol was able to successfully induce the expression of a gene controlled by Pzear, while estrone was only able to partially induce its expression but the other two compounds were not effective. In combination, these results demonstrate that ${\beta}$-estradiol can replace ZEA in this conditional gene expression system, thereby eliminating the need to use the more expensive reagent, ZEA, and facilitating high-throughput functional analyses of F. graminearum in future studies.

The Protein Kinase A Pathway Regulates Zearalenone Production by Modulating Alternative ZEB2 Transcription

  • Park, Ae Ran;Fu, Minmin;Shin, Ji Young;Son, Hokyoung;Lee, Yin-Won
    • Journal of Microbiology and Biotechnology
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    • v.26 no.5
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    • pp.967-974
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    • 2016
  • Zearalenone (ZEA) is an estrogenic mycotoxin that is produced by several Fusarium species, including Fusarium graminearum. One of the ZEA biosynthetic genes, ZEB2, encodes two isoforms of Zeb2 by alternative transcription, forming an activator (Zeb2L-Zeb2L homooligomer) and an inhibitor (Zeb2L-Zeb2S heterodimer) that directly regulate the ZEA biosynthetic genes in F. graminearum. Cyclic AMP-dependent protein kinase A (PKA) signaling regulates secondary metabolic processes in several filamentous fungi. In this study, we investigated the effects of the PKA signaling pathway on ZEA biosynthesis. Through functional analyses of PKA catalytic and regulatory subunits (CPKs and PKR), we found that the PKA pathway negatively regulates ZEA production. Genetic and biochemical evidence further demonstrated that the PKA pathway specifically represses ZEB2L transcription and also takes part in posttranscriptional regulation of ZEB2L during ZEA production. Our findings reveal the intriguing mechanism that the PKA pathway regulates secondary metabolite production by reprograming alternative transcription.

A comparison of functional MRI and MRSI on occipital cortex by visual stimulation (시각자극에 의한 후두엽 피질에서의 기능적 자기공명영상법과 양성자 대사물질영상의 비교)

  • Kim, T.;Suh, T.S.;Choe, B.Y.;Shinn, K.S.
    • Proceedings of the KOSOMBE Conference
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    • v.1998 no.11
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    • pp.291-292
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    • 1998
  • The purpose of the study was aimed to evaluate the BOLD contrast fMRI in occipital lobe and compare this imaging with metabolite changes based on $^1H$ MRS and MRSI before and after visual stimulation. As a result, the activation map were sucessfully produced by thresholding with minimum cross-correlate value of 0.45. In MRS, NAA/Cr ratio is almost same. however, latate was elevated almost 9 times higher than before activation. Lactate metabolic images were consistent with the BOLD effect map. The BOLD contrast fMRI is not enough to detect the activation area in human brain. so, the other modality was required such as lactate metabolic map.

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Proteomic Comparison of Gibberella moniliformis in Limited-Nitrogen (Fumonisin-Inducing) and Excess-Nitrogen (Fumonisin-Repressing) Conditions

  • Choi, Yoon-E;Butchko, Robert A.E.;Shim, Won-Bo
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.780-787
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    • 2012
  • The maize pathogen Gibberella moniliformis produces fumonisins, a group of mycotoxins associated with several disorders in animals and humans, including cancer. The current focus of our research is to understand the regulatory mechanisms involved in fumonisin biosynthesis. In this study, we employed a proteomics approach to identify novel genes involved in the fumonisin biosynthesis under nitrogen stress. The combination of genome sequence, mutant strains, EST database, microarrays, and proteomics offers an opportunity to advance our understanding of this process. We investigated the response of the G. moniliformis proteome in limited nitrogen (N0, fumonisin-inducing) and excess nitrogen (N+, fumonisin-repressing) conditions by one- and two-dimensional electrophoresis. We selected 11 differentially expressed proteins, six from limited nitrogen conditions and five from excess nitrogen conditions, and determined the sequences by peptide mass fingerprinting and MS/MS spectrophotometry. Subsequently, we identified the EST sequences corresponding to the proteins and studied their expression profiles in different culture conditions. Through the comparative analysis of gene and protein expression data, we identified three candidate genes for functional analysis and our results provided valuable clues regarding the regulatory mechanisms of fumonisin biosynthesis.