Metabolomics reveals mycoplasma contamination interferes with the metabolism of PANC-1 cells

Yu, Tao; Wang, Yong Tao; Zhang, Huizhen; Johnson, Caroline H.; Jiang, Yiming; Li, Xiangjun; Wu, Zeming; Liu, Tian; Krausz, Kristopher W.; Yu, Aiming; Gonzalez, Frank J.; Huang, Min; Bi, Huichang

doi:10.1007/s00216-016-9525-9

Cited by 17 publications

(9 citation statements)

References 34 publications

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“…Mycoplasma contamination is a common problem in cell culture and affects many aspects of cell physiology (Kagemann et al 2005a;Yu et al 2016). In this study, we found that mycoplasma contamination could increase the phosphorylation of NF-kB and MAPK signal pathway and induce the activation of BV2 cells.…”

Section: Discussionsupporting

confidence: 54%

Mycoplasma contamination affects cell characteristics and decreases the sensitivity of BV2 microglia to LPS stimulation

2019

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Mycoplasma is the most common contaminant and greatly affects host cells. The influence of mycoplasma on microglia cells remains unknown. Here, we investigated the influence of mycoplasma contamination on BV2 cells (a microglia cell line). We found that mycoplasma contamination increased the phosphorylation of NF-kB and MAPK signal pathway and induced the activation of BV2 cells. These mycoplasma-contaminated BV2 cells exhibited a transition of cell morphology and slower proliferation, as well as increased gene expression and protein secretion of inflammatory factors. Furthermore, mycoplasma-contaminated BV2 cells had decreased sensitivity to lipopolysaccharide stimulation. These findings suggested that mycoplasma contamination greatly influenced the characteristics and function of microglia cells. It is important to prevent and exclude mycoplasma contamination in our research.

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Section: Discussionsupporting

confidence: 54%

Mycoplasma contamination affects cell characteristics and decreases the sensitivity of BV2 microglia to LPS stimulation

2019

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“…The candidate markers were selected by examining the S-plot based on the variable importance (VIP) value, which was more than 1.0. The identification of the metabolites was confirmed by comparisons of fragmentation spectra and m/z through three main online databases: Metlin ( http://metlin.scripps.edu ), HMDB ( http://www.hmdb.ca/ ), and mzcloud ( https://www.mzcloud.org/ ) [ 14 , 15 ]. To assess the strength of association between individual metabolites and minimal/mild endometriosis, a step-wise logistic regression analysis with backward elimination was used to establish a model and filter crucial metabolites.…”

Section: Methodsmentioning

confidence: 99%

Endometrium metabolomic profiling reveals potential biomarkers for diagnosis of endometriosis at minimal-mild stages

Guan

Zhang

et al. 2018

Reprod Biol Endocrinol

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BackgroundThe sensitivity and specificity of non-invasive diagnostic methods for endometriosis, especially at early stages, are not optimal. The clinical diagnostic indicator cancer antigen 125 (CA125) performs poorly in the diagnosis of minimal endometriosis, with a sensitivity of 24%. Therefore, it is urgent to explore novel diagnostic biomarkers. We evaluated the metabolomic profile variation of the eutopic endometrium between minimal-mild endometriosis patients and healthy women by ultra-high-performance liquid chromatography coupled with electrospray ionization high-resolution mass spectrometry (UHPLC-ESI-HRMS).MethodsOur study comprised 29 patients with laparoscopically confirmed endometriosis at stages I-II and 37 infertile women who underwent diagnostic laparoscopy combined with hysteroscopy from January 2014 to January 2015. Eutopic endometrium samples were collected by pipelle endometrial biopsy. The metabolites were quantified by UHPLC-ESI-HRMS. The best combination of biomarkers was then selected by performing step-wise logistic regression analysis with backward elimination.ResultsTwelve metabolites were identified as endometriosis-associated biomarkers. The eutopic endometrium metabolomic profile of the endometriosis patients was characterized by a significant increase in the concentration of hypoxanthine, L-arginine, L-tyrosine, leucine, lysine, inosine, omega-3 arachidonic acid, guanosine, xanthosine, lysophosphatidylethanolamine and asparagine. In contrast, the concentration of uric acid was decreased. Metabolites were filtered by step-wise logistic regression with backward elimination, and a model containing uric acid, hypoxanthine, and lysophosphatidylethanolamine was constructed. Receiver-operating characteristic (ROC) analysis confirmed the prognostic value of these parameters for the diagnosis of minimal/mild endometriosis with a sensitivity of 66.7% and a specificity of 90.0%.ConclusionsMetabolomics analysis of the eutopic endometrium in endometriosis was effectively characterized by UHPLC-ESI-HRMS-based metabolomics. Our study supports the importance of purine and amino acid metabolites in the pathophysiology of endometriosis and provides potential biomarkers for semi-invasive diagnosis of early-stage endometriosis.

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“…Hypoxanthine-guanine phosphoribosyl transferase (HGPRT) and adenine phosphoribosyl transferase (APRT) can convert adenine, hypoxanthine and guanine to adenosine 5′-monophosphate (AMP), inosine 5′-phosphate (IMP) and guanosine 5′-phosphate (GMP), respectively. They are the substrates for the synthesis of high-energy nucleotides and it was reported that AMP and GMP levels were significantly up-regulated in normal gastric tissue from patients with gastric cancer [11,27,28]. In the present study, adenine, hypoxanthine and guanine were significantly upregulated in the chronic gastritis group, which might suggest that the synthesis of AMP, IMP, and GMP was enhanced.…”

Section: Purine Metabolism-supporting

confidence: 51%

Non-targeted metabolomics reveals diagnostic biomarker in the tongue coating of patients with chronic gastritis

Wang

et al. 2019

Journal of Pharmaceutical and Biomedical Analysis

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Analysis of the properties of the tongue has been used in traditional Chinese medicine for disease diagnosis. Notably, tongue analysis, which is non-invasive and convenient compared with gastroscopy and pathological examination, can be used to assess chronic gastritis (CG). In order to find potential diagnostic biomarkers and study the metabolic mechanisms of the endogenous small molecules in the tongue coating related to CG, a non-targeted metabolomic analysis method was developed using ultra high performance liquid chromatography combined with quadrupole timeof-flight mass spectrometry (UHPLC-Q/TOF-MS). It was performed using two different columns in positive and negative ion scanning modes separately. The stability of the samples was evaluated and the age and gender factors of the subjects were excluded to ensure the reliability of the data in this study. Finally, under the four analysis models, 130, 229, 113 and 92 differential compounds were found using multivariate statistical methods respectively. 37 potential biomarkers were putatively identified after removing the duplicate compounds and five potential diagnostic biomarkers were putatively identified by receiver operating characteristic (ROC) curve analysis, including inosine, oleamide, adenosine, N-acetylglucosamine (GlcNAc) and xanthine. The main metabolic pathways associated with CG were purine metabolism, amino acid metabolism, sphingolipid metabolism and energy metabolism, which suggested that oxygen free radicals and energy metabolism were altered in patients with CG. These results provided a potential new basis for the quantitative diagnosis and pathogenesis of CG.

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Metabolomics reveals mycoplasma contamination interferes with the metabolism of PANC-1 cells

Cited by 17 publications

References 34 publications

Mycoplasma contamination affects cell characteristics and decreases the sensitivity of BV2 microglia to LPS stimulation

Mycoplasma contamination affects cell characteristics and decreases the sensitivity of BV2 microglia to LPS stimulation

Endometrium metabolomic profiling reveals potential biomarkers for diagnosis of endometriosis at minimal-mild stages

Non-targeted metabolomics reveals diagnostic biomarker in the tongue coating of patients with chronic gastritis

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