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Graphical representation of the experimental setup. The experimental setup for the metabolic foot printing of E. multilocularis metacestodes in vitro included ( a ) medium preconditioning with rat hepatoma cells for four days in DMEM including 0.2% FCS to enrich the medium with host <t>cell</t> <t>metabolites</t> (cDMEM). ( b ) incubation of cDMEM with in vitro cultured E. multilocularis metacestode vesicles (vcDMEM) or mock incubation for control (ccDMEM) over 72 h (for 1 H <t>NMR</t> analysis) or for the time-points 0, 2, 6, 10, 24, 48, 72 h (for quantitative measurements). During this time, metacestode vesicles consumed and released metabolites (indicated by arrows). Samples of vcDMEM, ccDMEM, and vesicle fluid (VF) were harvested for subsequent metabolite identification by 1 H NMR, HPLC, or enzymatic assays.
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Graphical representation of the experimental setup. The experimental setup for the metabolic foot printing of E. multilocularis metacestodes in vitro included ( a ) medium preconditioning with rat hepatoma cells for four days in DMEM including 0.2% FCS to enrich the medium with host <t>cell</t> <t>metabolites</t> (cDMEM). ( b ) incubation of cDMEM with in vitro cultured E. multilocularis metacestode vesicles (vcDMEM) or mock incubation for control (ccDMEM) over 72 h (for 1 H <t>NMR</t> analysis) or for the time-points 0, 2, 6, 10, 24, 48, 72 h (for quantitative measurements). During this time, metacestode vesicles consumed and released metabolites (indicated by arrows). Samples of vcDMEM, ccDMEM, and vesicle fluid (VF) were harvested for subsequent metabolite identification by 1 H NMR, HPLC, or enzymatic assays.
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Graphical representation of the experimental setup. The experimental setup for the metabolic foot printing of E. multilocularis metacestodes in vitro included ( a ) medium preconditioning with rat hepatoma cells for four days in DMEM including 0.2% FCS to enrich the medium with host <t>cell</t> <t>metabolites</t> (cDMEM). ( b ) incubation of cDMEM with in vitro cultured E. multilocularis metacestode vesicles (vcDMEM) or mock incubation for control (ccDMEM) over 72 h (for 1 H <t>NMR</t> analysis) or for the time-points 0, 2, 6, 10, 24, 48, 72 h (for quantitative measurements). During this time, metacestode vesicles consumed and released metabolites (indicated by arrows). Samples of vcDMEM, ccDMEM, and vesicle fluid (VF) were harvested for subsequent metabolite identification by 1 H NMR, HPLC, or enzymatic assays.
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Graphical representation of the experimental setup. The experimental setup for the metabolic foot printing of E. multilocularis metacestodes in vitro included ( a ) medium preconditioning with rat hepatoma cells for four days in DMEM including 0.2% FCS to enrich the medium with host <t>cell</t> <t>metabolites</t> (cDMEM). ( b ) incubation of cDMEM with in vitro cultured E. multilocularis metacestode vesicles (vcDMEM) or mock incubation for control (ccDMEM) over 72 h (for 1 H <t>NMR</t> analysis) or for the time-points 0, 2, 6, 10, 24, 48, 72 h (for quantitative measurements). During this time, metacestode vesicles consumed and released metabolites (indicated by arrows). Samples of vcDMEM, ccDMEM, and vesicle fluid (VF) were harvested for subsequent metabolite identification by 1 H NMR, HPLC, or enzymatic assays.
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Graphical representation of the experimental setup. The experimental setup for the metabolic foot printing of E. multilocularis metacestodes in vitro included ( a ) medium preconditioning with rat hepatoma cells for four days in DMEM including 0.2% FCS to enrich the medium with host cell metabolites (cDMEM). ( b ) incubation of cDMEM with in vitro cultured E. multilocularis metacestode vesicles (vcDMEM) or mock incubation for control (ccDMEM) over 72 h (for 1 H NMR analysis) or for the time-points 0, 2, 6, 10, 24, 48, 72 h (for quantitative measurements). During this time, metacestode vesicles consumed and released metabolites (indicated by arrows). Samples of vcDMEM, ccDMEM, and vesicle fluid (VF) were harvested for subsequent metabolite identification by 1 H NMR, HPLC, or enzymatic assays.

Journal: Scientific Reports

Article Title: In vitro metabolomic footprint of the Echinococcus multilocularis metacestode

doi: 10.1038/s41598-019-56073-y

Figure Lengend Snippet: Graphical representation of the experimental setup. The experimental setup for the metabolic foot printing of E. multilocularis metacestodes in vitro included ( a ) medium preconditioning with rat hepatoma cells for four days in DMEM including 0.2% FCS to enrich the medium with host cell metabolites (cDMEM). ( b ) incubation of cDMEM with in vitro cultured E. multilocularis metacestode vesicles (vcDMEM) or mock incubation for control (ccDMEM) over 72 h (for 1 H NMR analysis) or for the time-points 0, 2, 6, 10, 24, 48, 72 h (for quantitative measurements). During this time, metacestode vesicles consumed and released metabolites (indicated by arrows). Samples of vcDMEM, ccDMEM, and vesicle fluid (VF) were harvested for subsequent metabolite identification by 1 H NMR, HPLC, or enzymatic assays.

Article Snippet: Metabolites were identified using Chenomx NMR Suite (V 8.2; May-01-2016 with Java 1.8.0_74 (x86)), Human Metabolite Database (HMDB) , 2D NMR spectra and the published 2D NMR data , , and statistical total correlation spectroscopy (STOCSY) with the script IMPaCTS (v 1.0.0) in Matlab (V R2015b 8.6.0.267245) .

Techniques: In Vitro, Incubation, Cell Culture

Multivariate statistical analysis of 1 H NMR spectra. Multivariate comparison between vcDMEM (red, n = 10) and ccDMEM (blue, n = 10) acquired from 1D 1 H NMR spectra after normalization. ( a ) Principal component analysis (PCA) scores plot of vcDMEM and ccDMEM. Groups are separated in the first principal component (PC1) with an explained variance of 98.39%. ( b ) Respective orthogonal projection to latent structure discriminant analysis (OPLS-DA) scores plot of vcDMEM and ccDMEM also clearly separated the two groups. Tcv depicts the inter-group variation, Tosc the intra-group variation. ( c ) Annotated PCA loadings plot of vcDMEM and ccDMEM. Annotated metabolites are indicated by various colors. ( d ) Annotated OPLS-DA coefficient plot of vcDMEM and ccDMEM. The x-axis depicts the chemical shifts in ppm. Peaks orientating upwards form the baseline show higher metabolite abundance in vcDMEM, from the baseline downwards pointing peaks show higher abundance in ccDMEM. The significance that each peak contributes to the difference between the two groups is indicated color-coded from blue low to red (scale on the right). The total explained variance ( R 2 X ) was 0.98 and the corresponding cross-validation ( Q 2 Y ) was 0.99. Amino acids are annotated using the three-letter code, for all other metabolites, full names are given.

Journal: Scientific Reports

Article Title: In vitro metabolomic footprint of the Echinococcus multilocularis metacestode

doi: 10.1038/s41598-019-56073-y

Figure Lengend Snippet: Multivariate statistical analysis of 1 H NMR spectra. Multivariate comparison between vcDMEM (red, n = 10) and ccDMEM (blue, n = 10) acquired from 1D 1 H NMR spectra after normalization. ( a ) Principal component analysis (PCA) scores plot of vcDMEM and ccDMEM. Groups are separated in the first principal component (PC1) with an explained variance of 98.39%. ( b ) Respective orthogonal projection to latent structure discriminant analysis (OPLS-DA) scores plot of vcDMEM and ccDMEM also clearly separated the two groups. Tcv depicts the inter-group variation, Tosc the intra-group variation. ( c ) Annotated PCA loadings plot of vcDMEM and ccDMEM. Annotated metabolites are indicated by various colors. ( d ) Annotated OPLS-DA coefficient plot of vcDMEM and ccDMEM. The x-axis depicts the chemical shifts in ppm. Peaks orientating upwards form the baseline show higher metabolite abundance in vcDMEM, from the baseline downwards pointing peaks show higher abundance in ccDMEM. The significance that each peak contributes to the difference between the two groups is indicated color-coded from blue low to red (scale on the right). The total explained variance ( R 2 X ) was 0.98 and the corresponding cross-validation ( Q 2 Y ) was 0.99. Amino acids are annotated using the three-letter code, for all other metabolites, full names are given.

Article Snippet: Metabolites were identified using Chenomx NMR Suite (V 8.2; May-01-2016 with Java 1.8.0_74 (x86)), Human Metabolite Database (HMDB) , 2D NMR spectra and the published 2D NMR data , , and statistical total correlation spectroscopy (STOCSY) with the script IMPaCTS (v 1.0.0) in Matlab (V R2015b 8.6.0.267245) .

Techniques:

Identified  Metabolites  in 1 H  NMR  spectra of vcDMEM, ccDMEM, and VF.

Journal: Scientific Reports

Article Title: In vitro metabolomic footprint of the Echinococcus multilocularis metacestode

doi: 10.1038/s41598-019-56073-y

Figure Lengend Snippet: Identified Metabolites in 1 H NMR spectra of vcDMEM, ccDMEM, and VF.

Article Snippet: Metabolites were identified using Chenomx NMR Suite (V 8.2; May-01-2016 with Java 1.8.0_74 (x86)), Human Metabolite Database (HMDB) , 2D NMR spectra and the published 2D NMR data , , and statistical total correlation spectroscopy (STOCSY) with the script IMPaCTS (v 1.0.0) in Matlab (V R2015b 8.6.0.267245) .

Techniques: