• Title/Summary/Keyword: Functional metabolite

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Study of Metabolic Profiling Changes in Colorectal Cancer Tissues Using 1D 1H HR-MAS NMR Spectroscopy

  • Kim, Siwon;Lee, Sangmi;Maeng, Young Hee;Chang, Weon Young;Hyun, Jin Won;Kim, Suhkmann
    • Bulletin of the Korean Chemical Society
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    • v.34 no.5
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    • pp.1467-1472
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    • 2013
  • Metabolomics is a field that studies systematic dynamics and secretion of metabolites from cells to understand biological pathways based on metabolite changes. The metabolic profiling of intact human colorectal tissues was performed using high-resolution magic angle spinning (HR-MAS) NMR spectroscopy, which was unnecessary to extract metabolites from tissues. We used two different groups of samples, which were defined as normal and cancer, from 9 patients with colorectal cancer and investigated the samples in NMR experiments with a water suppression pulse sequence. We applied target profiling and multivariative statistical analysis to the analyzed 1D NMR spectra to identify the metabolites and discriminate between normal and cancer tissues. Cancer tissue showed higher levels of arginine, betaine, glutamate, lysine, taurine and lower levels of glutamine, hypoxanthine, isoleucine, lactate, methionine, pyruvate, tyrosine relative to normal tissue. In the OPLS-DA (orthogonal partial least square discriminant analysis), the score plot showed good separation between the normal and cancer groups. These results suggest that metabolic profiling of colorectal cancer could provide new biomarkers.

In Vivo $^1H$ MR Spectroscopic Study on Levodopa-Treated Parkison's Disease

  • Choe, Bo-Young;Baik, Hyun-Man;Son, Byung-Chul;Kim, Moon-Chan;Kim, Euy-Neyung;Suh, Tae-Suk;Lee, Hyoung-Koo;Shinn, Kyung-Sub
    • Journal of the Korean Magnetic Resonance Society
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    • v.4 no.1
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    • pp.19-28
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    • 2000
  • Authors evaluated alterations of observable metabolite ratios between the cerebral lesion and the contralateral region related to the clinical symptomatic side in levodopa-treated Parkinson's disease (PD) and investigated correlation between age in patients with PD and metabolite ratios of the lesion. Patients with levodopa-treated PD (n = 54) and age-matched normal controls (n = 15) underwent magnetic resonance spectroscopy (MRS) examinations using a stimulated echo acquisition mode (STEAM) pulse sequence that provided 2$\times$2$\times$2 cm3 volume of interest in the selected regions of substantia nigra (SN) and putamed-globus pallidus (PG). To evaluate dependence of metabolite ratios on age, we divided into two groups (i.e., younger and older age). We quantitatively measured N-acetylaspartate (NAA), creatine (Cr), choline-containing compounds (Cho), inositols (Ins), and the sum of glutamate (Glx) and GABA levels and obtained proton metabolite ratios relative to Cr using a Marquart algorithm. Compared with the contralateral region, a significant neuronal laterality of the NAA/Cr ratio in the lesion of SN related to the clinical symptomatic side was established (P = 0.01), but was not established in the lesion of PG (P = 0.24). Also, Cho/Cr ratio tended toward significance in the lesion of SN (P = 0.07) and was statistically significant in the lesion of PG(P = 0.01). Compared with that in the younger age group, NAA/Cr ratio in the older age was decreased in the lesion of SN (P = 0.02), while NAA/Cr ratio was not statistically significant in the lesion of PG ( P = 0.21). Significant metabolic alterations of NAA/Cr and Cho/Cr ratios might be closely related with functional changes of neuropathological process in SN and PG of levodopa treated PD and could be a valuable finding for evaluation of the PD. A trend of NAA/Cr reduction, being statistically significant in older patients, could be indicative of more pronounced neuronal damage in the SN of the progressive PD.

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Assessment of Phenolic Content, Saponin Content, and Antioxidant Activities in Gray, Red, and White Adzuki Bean Germplasm: A Multivariate Analysis

  • Kebede Taye Desta;Hyemyeong Yoon;Myoung-Jae Shin;Sukyeung Lee;Xiaohan Wang;Yu-Mi Choi;Young-ah Jeon;YoungKwang Ju;JungYoon Yi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.68 no.3
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    • pp.147-166
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    • 2023
  • Seed color is controlled by several genes and is a key trait in determining the metabolite content and biological activities of legume genotypes. In this study, 296 adzuki bean accessions, including 159 grey, 99 red, and 38 white adzuki beans, were grown in Korea. Variations in total phenolic content (TPC), total saponin content (TSC), DPPH scavenging activity, ABTS•+ scavenging activity, and ferric reducing antioxidant power (FRAP) were assessed and were reported to be in the ranges of 1.52-8.24 mg GAE/g, 14.36-114.22 mg DE/g, 0.23-12.84 mg AAE/g, 1.05-17.66 mg TE/g, and 0.59-13.14 mg AAE/g, respectively, with a wide variation across adzuki beans. Except for DPPH scavenging activity, the average values declined in the order gray > red > white adzuki beans, each demonstrating a significant variation (p < 0.05). White adzuki beans, which showed low metabolite content and antioxidant activity, were clearly separated from the gray and red genotypes using principal component and hierarchical cluster analyses. Moreover, TPC, TSC, and antioxidant activities were strongly correlated, regardless of seed color. Overall, the diversity of the TPC, TSC, and antioxidant activity in a broad population of adzuki bean genotypes was determined. Furthermore, this study found that seed color variation in adzuki beans had a significant effect on the metabolite content and antioxidant activity. Superior accessions with high levels of TPC, TSC, and antioxidant activity were also discovered and could be used for functional plant breeding and human consumption. The findings of this study may be useful for understanding the relationship between seed coat color and metabolite concentration in adzuki beans, paving the way for molecular-level analyses.

Identification of Genes Associated with Fumonisin Biosynthesis in Fusarium verticillioides via Proteomics and Quantitative Real-Time PCR

  • Choi, Yoon-E.;Shim, Won-Bo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.648-657
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    • 2008
  • In this study, we used functional genomic strategies, proteomics and quantitative real-time (qRT)-PCR, to advance our understanding of genes associated with fumonisin production in the fungus Fusarium verticillioides. Earlier studies have demonstrated that deletion of the FCC1 gene, which encodes a C-type cyclin, leads to a drastic reduction in fumonisin production and conidiation in the mutant strain (FT536). The premise of our research was that comparative analysis of F. verticillioides wild-type and FT536 proteomes will reveal putative proteins, and ultimately corresponding genes, that are important for fumonisin biosynthesis. We isolated proteins that were significantly upregulated in either the wild type or FT536 via two-dimensional polyacrylamide gel electrophoresis, and subsequently obtained sequences by mass spectrometry. Homologs of identified proteins, e.g., carboxypeptidase, laccase, and nitrogen metabolite repression protein, are known to have functions involved in fungal secondary metabolism and development. We also identified gene sequences corresponding to the selected proteins and investigated their transcriptional profiles via quantitative real-time (qRT)-PCR in order to identify genes that show concomitant expression patterns during fumonisin biosynthesis. These genes can be selected as targets for functional analysis to further verify their roles in $FB_1$ biosynthesis.

Proteomic Reference Map and Comparative Analysis between Streptomyces griseus S4-7 and wbiE2 Transcription Factor-Mutant Strain

  • Kim, Jisu;Kwon, Young Sang;Bae, Dong-Won;Kwak, Youn-Sig
    • The Plant Pathology Journal
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    • v.36 no.2
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    • pp.185-191
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    • 2020
  • Streptomyces griseus S4-7, a well-characterized keystone taxon among strawberry microbial communities, shows exceptional disease-preventing ability. The whole-genome sequence, functional genes, and bioactive secondary metabolites of the strain have been described in previous studies. However, proteomics studies of not only the S4-7 strain, but also the Streptomyces genus as a whole, remain limited to date. Therefore, in the present study, we created a proteomics reference map for S. griseus S4-7. Additionally, analysis of differentially expressed proteins was performed against a wblE2 mutant, which was deficient in spore chain development and did not express an antifungal activity-regulatory transcription factor. We believe that our data provide a foundation for further in-depth studies of functional keystone taxa of the phytobiome and elucidation of the mechanisms underlying plant-microbe interactions, especially those involving the Streptomyces genus.

Wort Fermentation by Leuconostoc citreum Originated from Kimchi and Sensory Properties of Fermented Wort

  • Delgerzaya, Purev;Shin, Jin-Yeong;Kim, Kwang-Ok;Park, Jin-Byung
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1083-1090
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    • 2009
  • Fermentation of wort was investigated with an ultimate goal to develop a fermented beverage rich in prebiotics and functional ingredients as well as desirable in flavors. Wort was fermented with Leuconostoc citreum HJ-P4 originated from kimchi and subjected to sensory descriptive analysis. L. citreum HJ-P4 produced various organic acids (e.g., lactic acid, acetic acid) as well as functional sugars (e.g., mannitol, panose) during wort fermentation. The concentration and ratio of lactic acid and acetic acid were significantly influenced by roasting conditions of malts used for wort preparation and aeration conditions during fermentation. The concentration of mannitol and panose varied depending on the sucrose content of wort and aeration conditions. Sensory characteristics of the fermented worts were clearly differentiated according to the roasting conditions of malts used for wort preparation and aeration conditions during fermentation. These results indicate that metabolite concentration of fermented wort and its sensory properties can be manipulated with roasting conditions of malts and fermentation conditions.

Genetic Diversity and Metabolite Analysis of Gastrodia elata by Inter-Simple Sequence Repeats (ISSR) Markers (ISSR 표지에 의한 천마의 유전 다양성분석 및 기능성 물질분석)

  • Kim, Hyun Tae;Kim, Ji Ah;Park, Eung Jun
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.6
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    • pp.440-446
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    • 2012
  • Gastrodia elata, an achlorophyllous orchid plant, is rare medicinal plant. We investigated the genetic diversity in G. elata from 4 locations by using Inter-Simple Sequence Repeats (ISSR) markers. Shannon's information Index (S.I.) indicating genetic diversity ranged from 0.255 (Pocheon) to 0.322 (Muju) with the mean of 0.29. The level of genetic diversity was lower than other plant and most genetic diversity was allocated among individuals within populations (26.81%). The UPGMA dendrogram based on genetic distance failed in showing decisive geographic relationship. In the case of gastrodin (GA), the major components in G. elata, Sangju was highest. The ergothionine (ERG) was detected a lot of contents in Muju and Pocheon. In conclusion, our results is very important information for explaining relationship of genetic variation and functional substances without the effects of environment factors and developing genetic marker by ISSR in G. elata, which may be responsible for the development of breeds with a lot of functional substance in G. elata.

1D Proton NMR Spectroscopic Determination of Ethanol and Ethyl Glucuronide in Human Urine

  • Kim, Siwon;Lee, Minji;Yoon, Dahye;Lee, Dong-Kye;Choi, Hye-Jin;Kim, Suhkmann
    • Bulletin of the Korean Chemical Society
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    • v.34 no.8
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    • pp.2413-2418
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    • 2013
  • Forensic and legal medicine require reliable data to indicate excessive alcohol consumption. Ethanol is oxidatively metabolized to acetate by alcohol dehydrogenase and non-oxidatively metabolized to ethyl glucuronide (EtG), ethyl sulfate (EtS), phosphatidylethanol, or fatty acid ethyl esters (FAEE). Oxidative metabolism is too rapid to provide biomarkers for the detection of ethanol ingestion. However, the non-oxidative metabolite EtG is a useful biomarker because it is stable, non-volatile, water soluble, highly sensitive, and is detected in body fluid, hair, and tissues. EtG analysis methods such as mass spectroscopy, chromatography, or enzyme-linked immunosorbent assay techniques are currently in use. We suggest that nuclear magnetic resonance (NMR) spectroscopy could be used to monitor ethanol intake. As with current conventional methods, NMR spectroscopy doesn't require complicated pretreatments or sample separation. This method has the advantages of short acquisition time, simple sample preparation, reproducibility, and accuracy. In addition, all proton-containing compounds can be detected. In this study, we performed $^1H$ NMR analyses of urine to monitor the ethanol and EtG. Urinary samples were collected over time from 5 male volunteers. We confirmed that ethanol and EtG signals could be detected with NMR spectroscopy. Ethanol signals increased immediately upon alcohol intake, but decreased sharply over time. In contrast, EtG signal increased and reached a maximum about 9 h later, after which the EtG signal decreased gradually and remained detectable after 20-25 h. Based on these results, we suggest that $^1H$ NMR spectroscopy may be used to identify ethanol non-oxidative metabolites without the need for sample pretreatment.

Enhancement of ${\beta}$-Glucan Content in the Cultivation of Cauliflower Mushroom (Sparassis latifolia) by Elicitation

  • Park, Hyun;Ka, Kang-Hyeon;Ryu, Sung-Ryul
    • Mycobiology
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    • v.42 no.1
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    • pp.41-45
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    • 2014
  • The effectiveness of three kinds of enzymes (chitinase, ${\beta}$-glucuronidase, and lysing enzyme complex), employed as elicitors to enhance the ${\beta}$-glucan content in the sawdust-based cultivation of cauliflower mushroom (Sparassis latifolia), was examined. The elicitors were applied to the cauliflower mushroom after primordium formation, by spraying the enzyme solutions at three different levels on the sawdust-based medium. Mycelial growth was fully accomplished by the treatments, but the metabolic process during the growth of fruiting bodies was affected. The application of a lysing enzyme resulted in an increase in the ${\beta}$-glucan concentration by up to 31% compared to that of the control. However, the treatment resulted in a decrease in mushroom yield, which necessitated the need to evaluate its economic efficiency. Although we still need to develop a more efficient way for using elicitors to enhance functional metabolites in mushroom cultivation, the results indicate that the elicitation technique can be applied in the cultivation of medicinal/edible mushrooms.

Effect of Toluene Application to Skin on the Enhancement of Liver Injury in $CCl_4$-Pretreated Rats

  • Chae, Soon-Nim;Yoon, Chong-Guk;Cho, Hyun-Gug
    • Biomedical Science Letters
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    • v.7 no.2
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    • pp.79-83
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    • 2001
  • To evaluate an effect of toluene application to skin on the enhancement of liver damage in $CCl_4$-pretreated rats, toluene (35 mg/$cm^2$) was sequentially applied for 5 days to the skin of liver damaged rats with $CCl_4$ (6 times every other day: 0.1 ml/100 g body weight-50% $CCl_4$ in olive oil) On the basis of the functional and morphological findings in rat liver, appling toluene to the skin in liver damaged animals led to the more enhanced liver damage. In addition, by applying toluene to skin of liver damaged rats, the hepatic cytochrome P450 content was somewhat more increased, but the hepatic benzylalcohol dehydrogenase activity was significantly decreased (P<0.001), whereas benzaldehyde dehydrogenase activity was not statistically changed. In conclusion, the toluene application to skin in liver-damaged rat led to enhancement of liver injury that may be due to the accumulation of toluene metabolite in liver.

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