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Molecular characterization and quantification using state of the art solid-state adiabatic TOBSY NMR in burn trauma.

Righi V, Andronesi O, Mintzopoulos D, Tzika AA - Int. J. Mol. Med. (2009)

Bottom Line: Using 1- and 2-dimensional HRMAS NMR experiments, we identified several metabolites in burned tissues.Quantification of metabolites in burned tissues showed increased levels of lipid compounds, intracellular metabolites (e.g., taurine and phosphocreatine) and substantially decreased water-soluble metabolites (e.g., glutathione, carnosine, glucose, glutamine/glutamate and alanine).Moreover, this method has applications that facilitate the development of novel therapeutic strategies.

View Article: PubMed Central - PubMed

Affiliation: NMR Surgical Laboratory, Department of Surgery, Massachusetts General Hospital and Shriners Burn Institute, Harvard Medical School, Boston, MA 02114, USA.

ABSTRACT
We describe a novel solid-state nuclear magnetic resonance (NMR) method that maximizes the advantages of high-resolution magic-angle-spinning (HRMAS), relative conventional liquid-state NMR approaches, when applied to intact biopsies of skeletal muscle specimens collected from burn trauma patients. This novel method, termed optimized adiabatic TOtal through Bond correlation SpectroscopY (TOBSY) solid-state NMR pulse sequence for two-dimensional (2D) 1H-1H homonuclear scalar-coupling longitudinal isotropic mixing, was demonstrated to provide a 40-60% improvement in signal-to-noise ratio (SNR) relative to its liquid-state analogue TOCSY (TOtal Correlation SpectroscopY). Using 1- and 2-dimensional HRMAS NMR experiments, we identified several metabolites in burned tissues. Quantification of metabolites in burned tissues showed increased levels of lipid compounds, intracellular metabolites (e.g., taurine and phosphocreatine) and substantially decreased water-soluble metabolites (e.g., glutathione, carnosine, glucose, glutamine/glutamate and alanine). These findings demonstrate that HRMAS NMR Spectroscopy using TOBSY is a feasible technique that reveals new insights into the pathophysiology of burn trauma. Moreover, this method has applications that facilitate the development of novel therapeutic strategies.

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1D slices of acyl chain methylene (CH2)n and the relative spin system row for TOBSY (upper row) and TOCSY (lower panel).
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f5-ijmm-24-06-0749: 1D slices of acyl chain methylene (CH2)n and the relative spin system row for TOBSY (upper row) and TOCSY (lower panel).

Mentions: As illustrated in Fig. 4, 1D slices were extracted along the indirect dimension of 2D TOBSY (red) and 2D TOCSY (black) experiments that analyzed control skeletal muscle specimens to confirm that the transfer efficiency predicted for the TOBSY sequence was met. 1D slices corresponding to both small metabolites (i.e., Lac, Tau, HTau) and large molecules [i.e., vinyl protons of FA chains (CH═CH) and intramyocellular lipids (CH2)n at 5.33 and 1.33 ppm, respectively] are shown in Fig. 4 (slices scaled to a common noise level and peaks integrated). C9115 yielded higher signal intensities for low molecular weight metabolites (Fig. 4a), namely HTau and Tau (∼>50%), and Lac (∼>80%), as well as for high molecular weight metabolites (Fig. 4b), such as unsaturated acids (CH═CH) and intramyocellular lipids (CH2)n (∼>80%). An example of differential performance of the two 2D methods is presented in Fig. 5; note that the full FA (CH2)n spin system (six bonds), including the vinyl proton at 5.33 ppm (CH═CH), is apparent in the TOBSY spectrum but not the TOCSY spectrum.


Molecular characterization and quantification using state of the art solid-state adiabatic TOBSY NMR in burn trauma.

Righi V, Andronesi O, Mintzopoulos D, Tzika AA - Int. J. Mol. Med. (2009)

1D slices of acyl chain methylene (CH2)n and the relative spin system row for TOBSY (upper row) and TOCSY (lower panel).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3722686&req=5

f5-ijmm-24-06-0749: 1D slices of acyl chain methylene (CH2)n and the relative spin system row for TOBSY (upper row) and TOCSY (lower panel).
Mentions: As illustrated in Fig. 4, 1D slices were extracted along the indirect dimension of 2D TOBSY (red) and 2D TOCSY (black) experiments that analyzed control skeletal muscle specimens to confirm that the transfer efficiency predicted for the TOBSY sequence was met. 1D slices corresponding to both small metabolites (i.e., Lac, Tau, HTau) and large molecules [i.e., vinyl protons of FA chains (CH═CH) and intramyocellular lipids (CH2)n at 5.33 and 1.33 ppm, respectively] are shown in Fig. 4 (slices scaled to a common noise level and peaks integrated). C9115 yielded higher signal intensities for low molecular weight metabolites (Fig. 4a), namely HTau and Tau (∼>50%), and Lac (∼>80%), as well as for high molecular weight metabolites (Fig. 4b), such as unsaturated acids (CH═CH) and intramyocellular lipids (CH2)n (∼>80%). An example of differential performance of the two 2D methods is presented in Fig. 5; note that the full FA (CH2)n spin system (six bonds), including the vinyl proton at 5.33 ppm (CH═CH), is apparent in the TOBSY spectrum but not the TOCSY spectrum.

Bottom Line: Using 1- and 2-dimensional HRMAS NMR experiments, we identified several metabolites in burned tissues.Quantification of metabolites in burned tissues showed increased levels of lipid compounds, intracellular metabolites (e.g., taurine and phosphocreatine) and substantially decreased water-soluble metabolites (e.g., glutathione, carnosine, glucose, glutamine/glutamate and alanine).Moreover, this method has applications that facilitate the development of novel therapeutic strategies.

View Article: PubMed Central - PubMed

Affiliation: NMR Surgical Laboratory, Department of Surgery, Massachusetts General Hospital and Shriners Burn Institute, Harvard Medical School, Boston, MA 02114, USA.

ABSTRACT
We describe a novel solid-state nuclear magnetic resonance (NMR) method that maximizes the advantages of high-resolution magic-angle-spinning (HRMAS), relative conventional liquid-state NMR approaches, when applied to intact biopsies of skeletal muscle specimens collected from burn trauma patients. This novel method, termed optimized adiabatic TOtal through Bond correlation SpectroscopY (TOBSY) solid-state NMR pulse sequence for two-dimensional (2D) 1H-1H homonuclear scalar-coupling longitudinal isotropic mixing, was demonstrated to provide a 40-60% improvement in signal-to-noise ratio (SNR) relative to its liquid-state analogue TOCSY (TOtal Correlation SpectroscopY). Using 1- and 2-dimensional HRMAS NMR experiments, we identified several metabolites in burned tissues. Quantification of metabolites in burned tissues showed increased levels of lipid compounds, intracellular metabolites (e.g., taurine and phosphocreatine) and substantially decreased water-soluble metabolites (e.g., glutathione, carnosine, glucose, glutamine/glutamate and alanine). These findings demonstrate that HRMAS NMR Spectroscopy using TOBSY is a feasible technique that reveals new insights into the pathophysiology of burn trauma. Moreover, this method has applications that facilitate the development of novel therapeutic strategies.

Show MeSH
Related in: MedlinePlus