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Coffee is popular worldwide and its consumption is increasing in recent years. Although mass spectrometry-based lipidomics approaches have been prevalent, their application in studies related to detailed information and dynamic changes in lipid composition during coffee bean roasting is still limited. The aim of this study was to investigate the dynamic changes in coffee bean lipids during the roasting process. The lipid classes and lipid molecular species in coffee beans were characterized by lipidomic analysis combined with chemometrics. A total of 12 lipid classes and 105 lipid molecular species were identified and quantified. Triacylglycerols (TAG) was the most abundant lipid class in both green beans and roasted beans. The content of phosphatidylethanolamine (PE) and lysophosphatidylethanolamine (LPE) in green beans was obviously higher than that in roasted beans. Other phospholipids, such as phosphatidylinositol (PI), lysophosphatidylinositol (LPI), phosphatidylcholine (PC), lysophophatidylcholine (LPC) and phosphatidic acid (PA), showed a tendency to increase at the beginning of roasting, then decreased gradually. Several differential lipid molecule species, for instance, PE (16:0_18:2), PC (18:2_18:2) were significantly down-regulated, and PI (18:1_18:2) was significantly up-regulated. This study provided a scientific basis for the change of coffee bean lipids during the roasting process.

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Cooking methods affect the compositions of Lentinus edodes metabolites. Nevertheless, little information is available on the specific impact of different cooking methods on Lentinus edodes via metabolomic analysis. This study determined the influence of boiling, steaming, air-frying, and roasting on the metabolomic profiles of Lentinus edodes based on UHPLC-Q-Exactive Orbitrap MS/MS in combination with chemometrics. A total of 990 metabolites were detected and classified into 11 super-classes. Subsequently, the metabolites of the four cooking methods were distinguished using multivariate statistical analysis. The results showed that boiling caused a massive loss of metabolites while roasting and air-frying led to an evident upregulation. The upregulation of metabolites in the steaming groups was not as significant as in roasting and air-frying. This study provided reference data for a comprehensive understanding of the metabolites associated with domestic cooking methods and valuable guidance for the development of Lentinus edodes and its products in the future.

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In this study, ultra-high-performance liquid chromatography high-resolution accurate mass-mass spectrometry (UHPLC-HRAM/MS) was applied to characterize the lipid profiles of five crab species. A total of 203 lipid molecular species in muscle tissue and 176 in edible viscera were quantified. The results indicate that Cancer pagurus contained high levels of lipids with a docosahexaenoic acid (DHA) and eicosapntemacnioc acid (EPA) structure in the muscle tissue and edible viscera. A partial least squares discriminant analysis (PLS-DA) showed that PE 16:0/22:6, PE P-18:0/20:5, PA 16:0/22:6 and PC 16:0/16:1 could be used as potential biomarkers to discriminate the five kinds of crabs. In addition, some lipids, such as PE 18:0/20:5, PC 16:0/16:1, PE P-18:0/22:6 and SM 12:1;2O/20:0, could be used as characteristic molecules to distinguish between Cancer magister and Cancer pagurus, which are similar in appearance. This study provides a new perspective on discriminating crab species from MS-based lipidomics.

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Antarctic krill oil is functional oil and has a complex phospholipids composition that poses difficulties in elucidating its effect mechanism on ulcerative colitis (UC). The mechanism of UC action was studied by bioinformatics, and the therapeutic effect of Antarctic krill phospholipids (APL) on dextran sulfate sodium (DSS)-induced colitis mice was verified. GO functional enrichment analysis uncovered an enrichment of these genes in the regulation of cell-cell adhesion, membrane region, signaling receptor activator activity, and cytokine activity. Meanwhile, the KEGG results revealed the genes were enriched in the TNF signaling pathway, pathogenic Escherichia coli infection, inflammatory bowel disease and tight junction. Animal experiments showed that APL treatment alleviated the UC symptoms and reduced inflammatory damage. Meanwhile, the expressions of the tight junction (TJ) proteins, ZO-1 and occludin, were restored, and the levels of IL-6 and TNF-α were reduced. Moreover, Firmicutes/Bacteroidetes ratio in the intestinal microbiota was regulated, and the contents of short-chain fatty acids metabolites were raised. These findings would provide an insight for the beneficial effects of APL and dietary therapy strategies for UC.

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Chemiluminescence (CL) assay is a promising point-of-care testing (POCT) technology due to the fast response, high sensitivity, and easy miniaturization. The application and performance of CL POCT method were highly dependent on the CL reaction. Herein, based on the CL reaction between luminol and in-situ generated K3Fe(CN)6, a low-cost, enzyme-free, and label-free CL POCT method was explored via a portable and handheld luminometer to detect telomerase activity. Telomerase elongated telomere substrate (TS) primer to form (TTAGGG)n repeats which hybridize with multiple short DNAs. The intercalation of SYBR Green I (SGI) into double-stranded DNA (dsDNA) generated singlet oxygen under the irradiation of LED light source. Singlet oxygen was then employed for in-situ oxidation of K4Fe(CN)6 to K3Fe(CN)6, which could react with luminol to generate a strong CL intensity. Thus, telomerase activity could be specifically, sensitively, and label-free detected. The detection limit was down to 98 HeLa cells. The detection process was very simple, and the cost was about $0.01 for each measurement. Furthermore, telomerase activity was detectable in human serum samples, with spike recoveries from 96% to 105%. According to our knowledge, it is the first effort to develop a low-cost, label-free and enzyme-free CL method with good repeatability for detecting biomarker based on the analyte-triggered and in-situ generated K3Fe(CN)6/luminol CL reaction.

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The point of care testing (POCT) of telomerase activity is critical for early diagnosis of cancer. Herein, a colorimetric method was developed for visual detection of telomerase activity via hydrogen peroxide test strip. It is based on the telomerase-controlled in-situ formation of hydrogen peroxide. Firstly, biotinylated telomerase substrate (TS) primer was attached on the surface of magnetic beads (MBs) via the streptavidin-biotin reaction to form MB-TS complex. Then, TS primers were elongated by telomerase to form long telomere elongated products (TEP) which contains TTAGGG repeat units. The in-situ formed MB-TEP complex specifically hybridized with glucose oxidase modified cDNA (GOD-cDNA). After magnetic separation and washing, the MB-TEP/GOD-cDNA complex incubated with glucose solution to in-situ produce hydrogen peroxide which was detected by hydrogen peroxide test strip. One long TEP hybridized with multiple GOD-cDNAs, which enriched GOD to highly efficiently catalyze glucose for generating hydrogen peroxide. Thus, the visual assay achieved sensitive detection of telomerase activity, and the limit of detection (LOD) reached as low as 10 HeLa cells/µL by naked eyes and 4.5 HeLa cells/µL by absorbance measurements. Therefore, it offers a sensitive and low-cost method for visual detection of telomerase activity, which also, widens the application of commercial hydrogen peroxide test strip in the development of non-H2O2 biosensors.

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