The metabolic fingerprint of in vitro grown E. multilocularis metacestodes was determined. E. multilocularis metacestodes were grown in rat hepatoma-cell pre-conditioned culture medium (cDMEM) for 72 hours at 37°C. After this incubation period, parasite-interacted media (vcDMEM, n=10) and control incubated media (ccDMEM, n=10) were analyzed by nuclear magnetic resonance spectroscopy (NMR: 1D 1H, 2D 1H-1H, and 2D 1H-13C Bruker DRX 600MHz). In addition, also vesicle fluid (VF, n=5) samples from metacestodes were analyzed after 72 hours of incubation by NMR accordingly.
The names are corresponding to the .zip file in the archive
vcDMEM_ccDMEM-VF-1H-1D-NMR.zip 1D 1H NOESY
folder sample name
10 VF_1
20 VF_2
30 VF_3
40 VF_4
50 VF_5
610 vcDMEM_7
620 ccDMEM_3
640 vcDMEM_5
650 vcDMEM_6
680 ccDMEM_2
700 ccDMEM_9
720 vcDMEM_2
730 ccDMEM_1
740 ccDMEM_8
750 vcDMEM_1
800 ccDMEM_7
820 ccDMEM_4
850 vcDMEM_4
870 vcDMEM_8
900 vcDMEM_9
920 vcDMEM_3
970 vcDMEM_10
990 ccDMEM_6
1000 ccDMEM_5
1030 ccDMEM_10
vcDMEM_ccDMEM-COSY-STOCSY-HSQC-Jres-2D-NMR.zip 2D
folder sample name
10 ccDMEM NOESY
11 ccDMEM COSY
12 ccDMEM TOCSY
13 ccDMEM Jres
14 ccDMEM HSQC
20 vcDMEM NOESY
21 vcDMEM COSY
22 vcDMEM TOCSY
23 vcDMEM Jres
24 vcDMEM HSQC
VF-COSY-STOCSY-HSQC-Jres-2D-NMR.zip 2D
folder sample name
1 VF NOESY
2 VF COSY
3 VF TOCSY
4 VF Jres
5 VF HSQC
400 µl of each sample were mixed with 250 µl sodium phosphate buffer (D2O:H2O, v/v, 9:1; pH=7.4; 0.01% trimethylsilylpropanoic acid (TSP); 3 mM sodium-azide), vortexed and centrifuged. 600 µl of each sample were transferred into a 5 mm diameter NMR tube. 1H NMR spectroscopy was recorded at 300 K on a Bruker DRX 600MHz NMR spectrometer (Bruker Biospin; Rheinstetten, Germany) at 600.29 MHz. Each spectrum was acquired with 2.73 sec acquisition time for 128 scans (spectral width = 20.2 ppm) with a standard pulse sequence (recycle delay-90°−t1-90°-tm-90°-acquire free induction decay (FID)). Two-dimensional (2D) 1H-1H correlation spectroscopy (COSY), 1H-1H total correlation spectroscopy (TOCSY), and 1H-13C heteronuclear single quantum coherence (HSQC) NMR were acquired from selected samples from each sample group (32 scans).
