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Sample Preparation of Mycobacterium tuberculosis Extracts for Nuclear Magnetic Resonance Metabolomic Studies
Published on: September 3, 2012
Metabolites of alveolar Echinococcus as determined by [31P]- and [1H]-nuclear magnetic resonance spectroscopy
Abstract:
[31P]-Nuclear magnetic resonance (NMR) in vivo spectra of Echinococcus multilocularis cysts growing subcutaneously in Meriones unguiculatus showed prominent signals due to phosphomonoesters (PME), phosphodiesters (PDE), inorganic phosphate (Pi) and the alpha, beta and gamma phosphate groups of adenosine triphosphate (ATP). The internal pH of the parasite cysts was 6.7-6.8. The 31P spectra of extracts of these subcutaneous cysts showed peaks identified as glucose-6-phosphate (Glu-6-P), glycerol-3-phosphate (Gly-3-P), phosphorylethanolamine (PE), adenosine-5'-monophosphate (5'-AMP), nicotinamide adenine dinucleotide phosphate (NADP), phosphorylcholine (PC), Pi, glycerolphosphorylethanolamine (GPE), glycerolphosphorylcholine (GPC), phosphoenolpyruvate (PEP), adenosine diphosphate (ADP), ATP and diphosphodiesters (DPDE). These metabolites were also detected at comparable concentrations in the extracts of intraperitoneally grown cysts. In addition, significantly more phosphocreatine (PCr), probably of host origin, was detected in the subcutaneous cysts than in the intraperitoneal cysts. [1H]-NMR spectra of cyst extracts revealed that parasites grown in the abdominal cavity contained significantly less glucose but significantly more succinate, acetate, alanine and beta-hydroxybutyrate. Glycogen, creatine, glycine, taurine, betaine, cholines and lactate were present at similar concentrations in cyst material from both locations.
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