• Title/Summary/Keyword: metabolites analysis

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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.

Analyses of Expressed Sequence Tags from Chironomus riparius Using Pyrosequencing : Molecular Ecotoxicology Perspective

  • Nair, Prakash M. Gopalakrishnan;Park, Sun-Young;Choi, Jin-Hee
    • Environmental Analysis Health and Toxicology
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    • v.26
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    • pp.10.1-10.7
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    • 2011
  • Objects: Chironomus riparius, a non-biting midge (Chironomidae, Diptera), is extensively used as a model organism in aquatic ecotoxicological studies, and considering the potential of C. riparius larvae as a bio-monitoring species, little is known about its genome sequences. This study reports the results of an Expressed Sequence Tags (ESTs) sequencing project conducted on C. riparius larvae using 454 pyrosequencing. Method: To gain a better understanding of C. riparius transcriptome, we generated ESTs database of C.ripairus using pyrosequencing method. Results: Sequencing runs, using normalized cDNA collections from fourth instar larvae, yielded 20,020 expressed sequence tags, which were assembled into 8,565 contigs and 11,455 singletons. Sequence analysis was performed by BlastX search against the National Center for Biotechnology Information (NCBI) nucleotide (nr) and uniprot protein database. Based on the gene ontology classifications, 24% (E-value${\leq}1^{-5}$) of the sequences had known gene functions, 24% had unknown functions and 52% of sequences did not match any known sequences in the existing database. Sequence comparison revealed 81% of the genes have homologous genes among other insects belonging to the order Diptera providing tools for comparative genome analyses. Targeted searches using these annotations identified genes associated with essential metabolic pathways, signaling pathways, detoxification of toxic metabolites and stress response genes of ecotoxicological interest. Conclusions: The results obtained from this study would eventually make ecotoxicogenomics possible in a truly environmentally relevant species, such as, C. riparius.

Cloning and Characterization of Cinnamate-4-Hydroxylase Gene from Rubus occidentalis L.

  • Lee, Eun Mi;Lee, Seung Sik;An, Byung Chull;Barampuram, Shyamkumar;Kim, Jae-Sung;Cho, Jae-Young;Lee, In-Chul;Chung, Byung Yeoup
    • Journal of Radiation Industry
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    • v.2 no.3
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    • pp.97-104
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    • 2008
  • Cinnamate-4-hydroxylase (C4H) is a key enzyme of phenylpropanoid pathway, which leads a variety of secondary metabolites to participate in differentiation and protection of plant against environmental stresses. In this study, we isolated a full-length cDNA of the C4H gene from a black raspberry (Rubus occidentalis L.), using a reverse transcriptase-PCR and rapid amplification of the cDNA ends (RACE)-PCR. The full-length cDNA of the RocC4H gene contained a 1,515 bp open reading frame (ORF) encoding a 504 amino acid protein with a calculated molecular weight of about 57.9 kDa and an isoelectric point (pI) value of 9.1. The genomic DNA analysis revealed that RocC4H gene had three exons and two introns. By multiple sequence alignment, RocC4H protein was highly homologous with other plant C4Hs, and the cytochrome P450-featured motifs, such as the heme-binding domain, the T-containing binding pocket motif (AAIETT), the ERR triad, and the tetrapeptide (PPGP) hinge motif, were highly conserved. Southern blot analysis revealed that RocC4H is a single copy gene in R. occidentalis.

The Effects of Long-Term, Low-Level Exposure to Monocyclic Aromatic Hydrocarbons on Worker's Insulin Resistance

  • Won, Yong-Lim;Ko, Yong;Heo, Kyung-Hwa;Ko, Kyung-Sun;Lee, Mi-Young;Kim, Ki-Woong
    • Safety and Health at Work
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    • v.2 no.4
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    • pp.365-374
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    • 2011
  • Objectives: This study was designed to investigate whether long-term, low-level exposure to monocyclic aromatic hydrocarbons (MAHs) induced insulin resistance. Methods: The subjects were 110 male workers who were occupationally exposed to styrene, toluene, and xylene. One hundred and ten age-matched male workers who had never been occupationally exposed to organic solvents were selected as a control group. Cytokines, which have played a key role in the pathogenesis of insulin resistance, and oxidative stress indices were measured. Assessment of exposure to MAHs was performed by measuring their ambient levels and their urinary metabolites in exposed workers, and the resulting parameters between the exposed group and non-exposed control groups were compared. Results: There was no significant difference in general characteristics and anthropometric parameters between the two groups; however, total cholesterol, fasting glucose, fasting insulin, and homeostasis model assessment of insulin resistance levels were significantly higher in the exposed group. Phenylglyoxylic acid levels showed significant association with tumor necrosis factor-${\alpha}$, total oxidative status, and oxidative stress index via multiple linear regression analysis. Further, there was a negative correlation between methylhippuric acid levels and total anti-oxidative capacity, and there was a significant relationship between MAHs exposure and fasting glucose levels, as found by multiple logistic regression analysis (odds ratio = 3.95, 95% confidence interval = 1.074-14.530). Conclusion: This study indicated that MAHs increase fasting glucose level and insulin resistance. Furthermore, these results suggested that absorbing the organic solvent itself and active metabolic intermediates can increase oxidative stress and cytokine levels, resulting in the changes in glucose metabolism and the induction of insulin resistance.

Analyzing Correlations between Movie Characters Based on Deep Learning

  • Jin, Kyo Jun;Kim, Jong Wook
    • Journal of the Korea Society of Computer and Information
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    • v.26 no.10
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    • pp.9-17
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    • 2021
  • Humans are social animals that have gained information or social interaction through dialogue. In conversation, the mood of the word can change depending on the sensibility of one person to another. Relationships between characters in films are essential for understanding stories and lines between characters, but methods to extract this information from films have not been investigated. Therefore, we need a model that automatically analyzes the relationship aspects in the movie. In this paper, we propose a method to analyze the relationship between characters in the movie by utilizing deep learning techniques to measure the emotion of each character pair. The proposed method first extracts main characters from the movie script and finds the dialogue between the main characters. Then, to analyze the relationship between the main characters, it performs a sentiment analysis, weights them according to the positions of the metabolites in the entire time intervals and gathers their scores. Experimental results with real data sets demonstrate that the proposed scheme is able to effectively measure the emotional relationship between the main characters.

Genome-Based Insights into the Thermotolerant Adaptations of Neobacillus endophyticus BRMEA1T

  • Lingmin Jiang;Ho Le Han;Yuxin Peng;Doeun Jeon;Donghyun Cho;Cha Young Kim;Jiyoung Lee
    • Research in Plant Disease
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    • v.29 no.3
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    • pp.321-329
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    • 2023
  • The bacterium Neobacillus endophyticus BRMEA1T, isolated from the medicinal plant Selaginella involvens, known as its thermotolerant can grow at 50℃. To explore the genetic basis for its heat tolerance response and its potential for producing valuable natural compounds, the genomes of two thermotolerant and four mesophilic strains in the genus Neobacillus were analyzed using a bioinformatic software platform. The whole genome was annotated using RAST SEED and OrthVenn2, with a focus on identifying potential heattolerance-related genes. N. endophyticus BRMEA1T was found to possess more stress response genes compared to other mesophilic members of the genus, and it was the only strain that had genes for the synthesis of osmoregulated periplasmic glucans. This study sheds light on the potential value of N. endophyticus BRMEA1T, as it reveals the mechanism of heat resistance and the application of secondary metabolites produced by this bacterium through whole-genome sequencing and comparative analysis.

In Vitro Evaluation of Probiotic Properties of Two Novel Probiotic Mixtures, Consti-Biome and Sensi-Biome

  • You Jin Jang;Bonggyu Min;Jong Hyun Lim;Byung-Yong Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.9
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    • pp.1149-1161
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    • 2023
  • Changes in the gut microbiome cause recolonization by pathogens and inflammatory responses, leading to the development of intestinal disorders. Probiotics administration has been proposed for many years to reverse the intestinal dysbiosis and to enhance intestinal health. This study aimed to evaluate the inhibitory effects of two newly designed probiotic mixtures, Consti-Biome and Sensi-Biome, on two enteric pathogens Staphylococcus aureus and Escherichia coli that may cause intestinal disorders. Additionally, the study was designed to evaluate whether Consti-Biome and Sensi-Biome could modulate the immune response, produce short-chain fatty acids (SCFAs), and reduce gas production. Consti-Biome and Sensi-Biome showed superior adhesion ratios to HT-29 cells and competitively suppressed pathogen adhesion. Moreover, the probiotic mixtures decreased the levels of pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-6 and IL-1β. Cell-free supernatants (CFSs) were used to investigate the inhibitory effects of metabolites on growth and biofilms of pathogens. Consti-Biome and Sensi-Biome CFSs exhibited antimicrobial and anti-biofilm activity, where microscopic analysis confirmed an increase in the number of dead cells and the structural disruption of pathogens. Gas chromatographic analysis of the CFSs revealed their ability to produce SCFAs, including acetic, propionic, and butyric acid. SCFA secretion by probiotics may demonstrate their potential activities against pathogens and gut inflammation. In terms of intestinal symptoms regarding abdominal bloating and discomfort, Consti-Biome and Sensi-Biome also inhibited gas production. Thus, these two probiotic mixtures have great potential to be developed as dietary supplements to alleviate the intestinal disorders.

The Synergism of Human Lactobacillaceae and Inulin Decrease Hyperglycemia via Regulating the Composition of Gut Microbiota and Metabolic Profiles in db/db Mice

  • Peifan Li;Tong Tong;Yusong Wu;Xin Zhou;Michael Zhang;Jia Liu;Yongxin She;Zuming Li;Yongli Li
    • Journal of Microbiology and Biotechnology
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    • v.33 no.12
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    • pp.1657-1670
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    • 2023
  • This study aimed to evaluate the effects of Limosilactobacillus fermentum and Lactiplantibacillus plantarum isolated from human feces coordinating with inulin on the composition of gut microbiota and metabolic profiles in db/db mice. These supplements were administered to db/db mice for 12 weeks. The results showed that the Lactobacillaceae coordinating with inulin group (LI) exhibited lower fasting blood glucose levels than the model control group (MC). Additionally, LI was found to enhance colon tissue and increase the levels of short-chain fatty acids. 16S rRNA sequencing revealed that the abundance of Corynebacterium and Proteus, which were significantly increased in the MC group compared with NC group, were significantly decreased by the treatment of LI that also restored the key genera of the Lachnospiraceae_NK4A136_group, Lachnoclostridium, Ruminococcus_gnavus_group, Desulfovibrio, and Lachnospiraceae_UCG-006. Untargeted metabolomics analysis showed that lotaustralin, 5-hydroxyindoleacetic acid, and 13(S)-HpODE were increased while L-phenylalanine and L-tryptophan were decreased in the MC group compared with the NC group. However, the intervention of LI reversed the levels of these metabolites in the intestine. Correlation analysis revealed that Lachnoclostridium and Ruminococcus_gnavus_group were negatively correlated with 5-hydroxyindoleacetic acid and 13(S)-HpODE, but positively correlated with L-tryptophan. 13(S)-HpODE was involved in the "linoleic acid metabolism". L-tryptophan and 5-hydroxyindoleacetic acid were involved in "tryptophan metabolism" and "serotonergic synapse". These findings suggest that LI may alleviate type 2 diabetes symptoms by modulating the abundance of Ruminococcus_gnavus_group and Lachnoclostridium to regulate the pathways of "linoleic acid metabolism", "serotonergic synapse", and" tryptophan metabolism". Our results provide new insights into prevention and treatment of type 2 diabetes.

Gestational Exposure to Bisphenol A Causes DNA Hypomethylation and the Upregulation of Progesterone Receptor Expression in the Uterus in Adult Female Offspring Rats

  • Seung Gee Lee;Ji-Eun Park;Yong-Pil Cheon;Jong-Min Kim
    • Development and Reproduction
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    • v.27 no.4
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    • pp.195-203
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    • 2023
  • Exposure to environmental chemicals, including endocrine-disrupting chemicals, during the gestational period can have profound adverse effects on several organs in offspring. Bisphenol A (BPA) can infiltrate the human body through food and drinks, and its metabolites can cross both the placental and the blood-brain barriers. In this study, we investigate the effect of gestational exposure to BPA on epigenetic, biochemical, and histological modifications in the uterine tissues of F1 adult offspring rats. Pregnant rats were exposed to BPA from gestational day 8-15, and changes in global DNA methylation in uterine tissues obtained from adult offspring born to the exposed mothers were analyzed. Global DNA methylation analysis revealed that gestational exposure to BPA resulted in DNA hypomethylation in the uterus. Progesterone receptor (PR) protein expression in uterine tissues was monitored using western blot analysis, which revealed that the PR protein content was considerably higher in all BPA-exposed groups than in the control. Immunohistochemical examination for the PR revealed that intense PR-positive cells were more frequently observed in the BPA-exposed group than in the control group. To date, the evidence that the upregulation of PRs observed in the present study was caused by the non-methylation of specific PR promoter regions is lacking. Conclusively, these results indicate that exposure to BPA during gestation induces epigenetic alterations in the uteri of adult female offspring. We speculate that the global DNA hypomethylation and upregulation of the PR observed simultaneously in this study might be associated with the uterus.

Microcystins and Nodularin in Agricultural Products: Toxicity, Analytical Methods, Contamination Pathway, Occurrence, and Safety Management (농산물 내 마이크로시스틴과 노둘라린: 독성, 분석법, 오염 경로, 오염 현황 및 관리 동향)

  • Su Been Park;Sang Yoo Lee;Ji Eun Park;Jae Sung Kim;Hyang Sook Chun
    • Journal of Food Hygiene and Safety
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    • v.39 no.3
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    • pp.191-208
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    • 2024
  • The peptide-type hepatotoxins microcystins (MCs) and nodularin (NOD) are secondary metabolites produced by cyanobacteria. MCs and NOD can bioaccumulate in agricultural products through toxin-contaminated water, soil, and manure and can cause human health risks through the consumption of agricultural products. As interest in the contamination of agricultural products by MCs or NOD has recently emerged, occurrence studies based on various analysis methods for agricultural products have been conducted. However, studies on agricultural products are still insufficient compared to research on drinking water and seafood. In addition, research is primarily conducted on agricultural products grown in areas where green algae occur, but not on marketed products. In the present study, we review the physicochemical properties, toxicity, analysis methods, occurrence studies, and management status of MCs and NOD in agricultural products to build a foundation for systematic monitoring and safety management.