• Title/Summary/Keyword: metabolite analysis

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1H NMR metabolomics study for diabetic neuropathy and diabetes

  • Hyun, Ja-Shil;Yang, Jiwon;Kim, Hyun-Hwi;Lee, Yeong-Bae;Park, Sung Jean
    • Journal of the Korean Magnetic Resonance Society
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    • v.22 no.4
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    • pp.149-157
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    • 2018
  • Diabetes is known to be one of common causes for several types of peripheral nerve damage. Diabetic neuropathy (DN) is a significant complication lowering the quality of life that can be frequently found in diabetes patients. In this study, the metabolomic characteristic of DN and Diabetes was investigated with NMR spectroscopy. The sera samples were collected from DN patients, Diabetes patients, and healthy volunteers. Based on the pair-wise comparison, three metabolites were found to be noticeable: glucose, obviously, was upregulated both in DN patients (DNP) and Diabetes. Citrate is also increased in both diseases. However, the dietary nutrient and biosynthesized metabolite from glucose, ascorbate, was elevated only in DNP, compared to healthy control. The multivariate model of OPLS-DA clearly showed the group separation between healthy control-DNP and healthy control-Diabetes. The most significant metabolites that contributed the group separation included glucose, citrate, ascorbate, and lactate. Lactate did not show the statistical significance of change in t-test while it tends to down-regulated both in DNP and Diabetes. We also conducted the ROC curve analysis to make a multivariate model for discrimination of healthy control and diseases with the identified three metabolites. As a result, the discrimination model between healthy control and DNP (or Diabetes) was successful while the model between DNP and Diabetes was not satisfactory for discrimination. In addition, multiple combinations of lactate and citrate in the OPLS-DA model of healthy control and diabetes group (DNP + Diabetes patients) gave good ROC value of 0.952, which imply these two metabolites could be used for diagnosis of Diabetes without glucose information.

Draft Genome Analysis of Antimicrobial Streptomyces Isolated from Himalayan Lichen

  • Kim, Byeollee;Han, So-Ra;Lamichhane, Janardan;Park, Hyun;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1144-1154
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    • 2019
  • There have been several studies regarding lichen-associated bacteria obtained from diverse environments. Our screening process identified 49 bacterial species in two lichens from the Himalayas: 17 species of Actinobacteria, 19 species of Firmicutes, and 13 species of Proteobacteria. We discovered five types of strong antimicrobial agent-producing bacteria. Although some strains exhibited weak antimicrobial activity, NP088, NP131, NP132, NP134, and NP160 exhibited strong antimicrobial activity against all multidrug-resistant strains. Polyketide synthase (PKS) fingerprinting revealed results for 69 of 148 strains; these had similar genes, such as fatty acid-related PKS, adenylation domain genes, PfaA, and PksD. Although the association between antimicrobial activity and the PKS fingerprinting results is poorly resolved, NP160 had six types of PKS fingerprinting genes, as well as strong antimicrobial activity. Therefore, we sequenced the draft genome of strain NP160, and predicted its secondary metabolism using antiSMASH version 4.2. NP160 had 46 clusters and was predicted to produce similar secondary metabolites with similarities of 5-100%. Although NP160 had 100% similarity with the alkylresorcinol biosynthetic gene cluster, our results showed low similarity with existing members of this biosynthetic gene cluster, and most have not yet been revealed. In conclusion, we expect that lichen-associated bacteria from the Himalayas can produce new secondary metabolites, and we found several secondary metabolite-related biosynthetic gene clusters to support this hypothesis.

Complete genome sequencing of Pseudomonas fluorescens NBC275, a biocontrol agent against fungal pathogens of plants and insects (식물 및 곤충의 곰팡이 병원균에 항균력을 가진 Pseudomonas fluorescens NBC275 균주의 유전체 염기서열)

  • Dutta, Swarnalee;Yu, Sang-Mi;Nagendran, Rajalingam;Jeong, Sang Chul;Lee, Yong Hoon
    • Korean Journal of Microbiology
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    • v.55 no.2
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    • pp.157-159
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    • 2019
  • Pseudomonas fluorescens NBC275 (Pf275) isolated from soil sample collected at riverside of Nakdonggang showed antagonistic activity against fungal pathogens of plants and insects. Here we present complete genome sequence of Pf275. The genome comprises of 6,610,362 bp with GC content of 60.9%, which includes 5,869 predicted protein-coding genes, 16 rRNAs, and 65 tRNAs. Genome analysis revealed gene clusters encoding antimicrobial secondary metabolites such as pyoverdine, 2, 4-diacetylphloroglucinol, and phenazine, which are known to play essential roles in biocontrol of diseases.

Metabolome Analysis and Aroma Characteristics of Fermented Fruit Vinegar (발효 과일식초의 대사체 분석 및 향기 특성)

  • Choi, Chan-Yeong;Park, Eun-Hee;Ryu, Su-Jin;Shin, Woo-Chang;Kim, Myoung-Dong
    • Microbiology and Biotechnology Letters
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    • v.46 no.4
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    • pp.416-424
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    • 2018
  • Vinegar was prepared from the fruits produced in Gangwon province, and major metabolite and aroma components were investigated for acetic acid fermentation. In the case of Meoru-Bokbunja vinegar, the ${\text\tiny{L}}$-alanine content was greatly changed by acetic acid fermentation. Acetic acid had the highest content (43%) of total aromatic components, and the contents of ester compounds, such as ethyl acetate and isoamyl acetate, were significantly increased after fermentation. Omija-Makgeolli vinegar produced linalool and hexanoic acid by fermentation, and terpenoid compound was prevalent (41.5%). ${\text\tiny{L}}$-alanine was also increased in Omija-Makgeolli vinegar, similar to that of Meoru-Bokbunja vinegar. Terpene compounds, such as terpinel-4-ol and ${\alpha}$-terpineol in Omija-Makgeolli vinegar, and ethyl acetate in Meoru-Bokbunja vinegar, were identified as major components in each aromatic formulation.

Method validation of detecting ethanol metabolites (EtG, EtS) in post-mortem spleen (비장 조직에서 에탄올 대사체(EtG, EtS)를 검출하는 방법과 유효성 확인)

  • Kim, Soo-Min;Jo, Young-Hoon;An, Song-Hie;Lee, Woo-Jae;Kwon, Mia
    • Analytical Science and Technology
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    • v.34 no.3
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    • pp.115-121
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    • 2021
  • Ethyl glucuronide (EtG) and ethyl sulfate (EtS), which are ethanol metabolites, are direct indicators of ethanol intake; they have been studied in a variety of biological samples in forensic science. It is necessary to analyze ethanol metabolites to determine whether the ethanol detected in autopsy cases was due to alcohol consumption before death or due to the ethanol produced from post-mortem decay. In general, EtG and EtS are detected in the blood together with ethanol; however, it may be difficult to secure blood depending on the extent of decay. Therefore, the aforementioned method should be replaced by detecting the ethanol metabolites using tissue biological samples. In this study, we determined the optimal experimental conditions for detecting EtG and EtS from spleen samples using Liquid Chromatography - Tandem Mass Spectrometry (LC-MS/MS). Herein, the test method was validated, and an analysis method was applied to the actual autopsy cases.

Acremonidin E produced by Penicillium sp. SNF123, a fungal endophyte of Panax ginseng, has antimelanogenic activities

  • Kim, Kyuri;Jeong, Hae-In;Yang, Inho;Nam, Sang-Jip;Lim, Kyung-Min
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.98-107
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    • 2021
  • Background: Ginseng extracts and ginseng-fermented products are widely used as functional cosmetic ingredients for their whitening and antiwrinkle effects. Recently, increasing attention has been given to bioactive metabolites isolated from endophytic fungi. However, little is known about the bioactive metabolites of the fungi associated with Panax ginseng Meyer. Methods: An endophytic fungus, Penicillium sp. SNF123 was isolated from the root of P. ginseng, from which acremonidin E was purified. Acremonidin E was tested on melanin synthesis in the murine melanoma cell line B16F10, in the human melanoma cell line MNT-1, and in a pigmented 3D-human skin model, Melanoderm. Results: Acremonidin E reduced melanogenesis in α-melanocyte-stimulating hormone (α-MSH)-stimulated B16F10 cells with minimal cytotoxicity. qRT-PCR analysis demonstrated that acremonidin E downregulated melanogenic genes, including tyrosinase and tyrosinase-related protein 1 (TRP-1), while their enzymatic activities were unaffected. The antimelanogenic effects of acremonidin E were further confirmed in MNT-1 and a pigmented 3D human epidermal skin model, Melanoderm. Immunohistological examination of the Melanoderm further confirmed the regression of both melanin synthesis and melanocyte activation in the treated tissue. Conclusion: This study demonstrates that acremonidin E, a bioactive metabolite derived from a fungal endophyte of P. ginseng, can inhibit melanin synthesis by downregulating tyrosinase, illuminating the potential utility of microorganisms associated with P. ginseng for cosmetic ingredients.

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.

Nrf2-mediated activation of HO-1 is required in the blocking effect of compound K, a ginseng saponin metabolite, against oxidative stress damage in ARPE-19 human retinal pigment epithelial cells

  • Cheol Park;Hee-Jae Cha;Kyoung-Seob Song;Heui-Soo Kim;EunJin Bang;Hyesook Lee;Cheng-Yun Jin;Gi-Young Kim;Yung Hyun Choi
    • Journal of Ginseng Research
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    • v.47 no.2
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    • pp.311-318
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    • 2023
  • Background: The beneficial effects of compound K (CK) on different chronic diseases have been shown to be at least related to antioxidant action. Nevertheless, since its antioxidant activity in human retinal pigment epithelial (RPE) cells is still unknown, here we investigated whether CK alleviates oxidative stress-stimulated damage in RPE ARPE-19 cells. Methods: The cytoprotective consequence of CK in hydrogen peroxide (H2O2)-treated cells was evaluated by cell viability, DNA damage, and apoptosis assays. Fluorescence analysis and immunoblotting were performed to investigate the inhibitory action of CK on reactive oxygen species (ROS) production and mitochondrial dysfunction. Results: H2O2-promoted cytotoxicity, oxidative stress, DNA damage, mitochondrial impairment, and apoptosis were significantly attenuated by CK in ARPE-19 cells. Furthermore, nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation level and its shuttling to the nucleus were increased, which was correlated with upregulated activation of heme oxygenase-1 (HO-1). However, zinc protoporphyrin, a blocker of HO-1, significantly abrogated the preventive action of CK in H2O2-treated ARPE-19 cells. Conclusion: This study indicates that activation of Nrf2/HO-1 signaling by CK plays an important role in rescuing ARPE-19 cells from oxidative cellular damage.

Multi-Function of a New Bioactive Secondary Metabolite Derived from Endophytic Fungus Colletotrichum acutatum of Angelica sinensis

  • Ramy S. Yehia
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.806-822
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    • 2023
  • In the current study we assessed a new crystallized compound, 5-(1-hydroxybutyl)-4-methoxy-3-methyl-2H-pyran-2-one (C-HMMP), from the endophytic fungus Colletotrichum acutatum residing in the medicinal plant Angelica sinensis for its in vitro antimicrobial, antibiofilm, antioxidant, antimalarial, and anti-proliferative properties. The promising compound was identified as C-HMMP through antimicrobial-guided fraction. The structure of C-HMMP was unambiguously confirmed by 2D NMR and HIRS spectroscopic analysis. Antimicrobial property testing of C-HMMP showed it to be effective against a variety of pathogenic bacteria and fungi with MICs ranging from 3.9 to 31.25 ㎍/ml. The compound displayed excellent antibiofilm activity against C. albicans, S. aureus, and K. pneumonia. Furthermore, the antimalarial and radical scavenging activities of C-HMMP were clearly dosedependent, with IC50 values of 0.15 and 131.2 ㎍/ml. The anti-proliferative activity of C-HMMP against the HepG-2, HeLa, and MCF-7 cell lines in vitro was investigated by MTT assay, revealing notable anti-proliferative activity with IC50 values of 114.1, 90, and 133.6 ㎍/ml, respectively. Moreover, CHMMP successfully targets topoisomerase I and demonstrated beneficial anti-mutagenicity in the Ames test against the reactive carcinogenic mutagen, 2-aminofluorene (2-AF). Finally, the compound inhibited the activity of α-glucosidase and α-amylase with IC50 values of 144.7 and 118.6 ㎍/ml, respectively. To the best of our knowledge, the identified compound C-HMMP was obtained for the first time from C. acutatum of A. sinensis, and this study demonstrated that C-HMMP has relevant biological significance and could provide better therapeutic targets against disease.

A Pilot Study Exploring Temporal Development of Gut Microbiome/Metabolome in Breastfed Neonates during the First Week of Life

  • Imad Awan;Emily Schultz;John D. Sterrett;Lamya'a M. Dawud;Lyanna R. Kessler;Deborah Schoch;Christopher A. Lowry;Lori Feldman-Winter;Sangita Phadtare
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.26 no.2
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    • pp.99-115
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    • 2023
  • Purpose: Exclusive breastfeeding promotes gut microbial compositions associated with lower rates of metabolic and autoimmune diseases. Its cessation is implicated in increased microbiome-metabolome discordance, suggesting a vulnerability to dietary changes. Formula supplementation is common within our low-income, ethnic-minority community. We studied exclusively breastfed (EBF) neonates' early microbiome-metabolome coupling in efforts to build foundational knowledge needed to target this inequality. Methods: Maternal surveys and stool samples from seven EBF neonates at first transitional stool (0-24 hours), discharge (30-48 hours), and at first appointment (days 3-5) were collected. Survey included demographics, feeding method, medications, medical history and tobacco and alcohol use. Stool samples were processed for 16S rRNA gene sequencing and lipid analysis by gas chromatography-mass spectrometry. Alpha and beta diversity analyses and Procrustes randomization for associations were carried out. Results: Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria were the most abundant taxa. Variation in microbiome composition was greater between individuals than within (p=0.001). Palmitic, oleic, stearic, and linoleic acids were the most abundant lipids. Variation in lipid composition was greater between individuals than within (p=0.040). Multivariate composition of the metabolome, but not microbiome, correlated with time (p=0.030). Total lipids, saturated lipids, and unsaturated lipids concentrations increased over time (p=0.012, p=0.008, p=0.023). Alpha diversity did not correlate with time (p=0.403). Microbiome composition was not associated with each samples' metabolome (p=0.450). Conclusion: Neonate gut microbiomes were unique to each neonate; respective metabolome profiles demonstrated generalizable temporal developments. The overall variability suggests potential interplay between influences including maternal breastmilk composition, amount consumed and living environment.