Evidence for bi-exponential transverse relaxation of lactate in excised rat muscle.

To elucidate the low proton nuclear magnetic resonance (NMR) visibility of muscle lactate previously demonstrated in excised rat muscle, lactate transverse relaxation was investigated in the same model using double quantum editing sequences with effective echo times ranging from 55 to 475 msec. On this time scale, muscle lactate clearly exhibits a bi-exponential transverse relaxation with a short T2 of 33+/-5 msec (mean +/- SE, n = 3) and a long T2 of 230+/-10 msec. The relative populations (84+/-4% vs. 16+/-4%, respectively) of these two lactate pools are compatible with compartmentation between intra- and extracellular muscle lactate.

Low visibility of lactate in excised rat muscle using double quantum proton spectroscopy.

Lactate NMR visibility was investigated in excised rat muscle at 3 T by comparing the concentration determined in situ by double quantum (DQ) proton spectroscopy (150 ms effective echo time) to the concentration measured in vitro from perchloric acid extracts of the same muscle samples. After 1-2 h of ischemia, lactate NMR visibility was 32 +/- 3% (+/- SE, n = 9), and was only 21 +/- 1% (n = 6) after 10-12 h. Muscle lactate T2 was 140 +/- 11 ms and 184 +/- 6 ms, respectively. All potential mechanisms of DQ lactate signal attenuation (B0 and B1 inhomogeneity, DQ transverse relaxation, diffusion) were examined, and accounted for when necessary. A significant increase in lactate NMR visibility was demonstrated using a shorter effective echo time (79 ms) DQ editing sequence. These results are interpreted as reflecting muscle lactate compartmentation between a long T2 pool predominantly detected by DQ spectroscopy, and a short T2 pool virtually invisible with longer echo time NMR techniques.

An interleaved heteronuclear NMRI-NMRS approach to non-invasive investigation of exercising human skeletal muscle.

Novel tools are presented that aim at more comprehensive NMR investigations of human skeletal muscle metabolism, in particular during exercise protocols. They integrate imaging (NMRI) and spectroscopy (NMRS) experiments in a single dynamic examination. The first sequence that we propose combine NMR-plethysmography, 1H-NMRS of deoxymyoglobin and 31P-NMRS. This allows simultaneous determination of skeletal muscle perfusion, oxygenation and high-energy phosphates status. It is very well suited to the study of interplay between blood supply and energy metabolism during the recovery period from aerobic or anaerobic exercise. In a second sequence, the same spectroscopic measurements are associated to a 1H double quantum coherence (DQC) edition of lactate. It is, this time, possible to estimate muscle lactate production concurrently with oxygen content, high-energy phosphates distribution and intracellular pH. This sequence is intended mainly for metabolic investigations of ischemic bouts. Examples are given of the use of these sequences in normal adult volunteers. They demonstrate the technical feasibility of these new approaches and illustrate their potential for future applications, particularly non-invasive of regulatory mechanisms of muscle metabolism in situ.

Simultaneous determination of muscle perfusion and oxygenation by interleaved NMR plethysmography and deoxymyoglobin spectroscopy.

A novel approach is presented that combines NMR-plethysmography and NMRS of deoxymyoglobin in real-time, using line-by-line interleaved acquisitions of both gradient echo images during venous occlusion and of the N-delta proton signal of myoglobin's proximal F8 histidine. This method allowed simultaneous measurement of peripheral regional perfusion and skeletal muscle oxygen content. During reactive hyperaemia, using our combined NMRI-NMRS protocol, we explored the relationship between muscle reoxygenation (myoglobin resaturation half-time, y in s) and reperfusion (x in ml/100 g tissue/min) and found it to be highly significant (y = 70.83x-0.94; r2 = 0.70; F = 64.40; p = 9.73 x 10(-9). We also demonstrated that at low flow, muscle perfusion was a rate-limiting factor to reoxygenation. Making certain hypotheses, muscle oxygen extraction was derived from perfusion and myoglobin resaturation rate. Muscle oxygen extraction during early post-ischemic recovery (0.78 +/- 0.11, 0.79 +/- 0.09 and 0.72 +/- 0.05 at 0, 60 and 100 Torr counter-pressure, respectively) was shown to be independent of perfusion and maximum at each step of the protocol in most volunteers but also to display significant variability among subjects in this supposedly normal population sample.

Practical implementation of single-voxel double-quantum editing on a whole-body NMR spectrometer: localized monitoring of lactate in the human leg during and after exercise.

The classical double-quantum editing sequence 90 degrees,x-tau-180 degrees y-tau-90 degrees x-t1-90 degrees x-tau-180 degrees y-tau-AQ (tau = 1/4J) was rendered volume selective, by making slice selective the first 90 degrees pulse and the two 180 degrees pulses. Using simple rules to ensure optimum radio frequency phase coherence, this single-voxel editing sequence, reminiscent of a basic PRESS localization technique, was implemented on a whole-body 3 T spectrometer, and in vitro editing of lactate methyl protons was demonstrated without any significant loss in intrinsic sensitivity. The effectiveness of the proposed approach in vivo was also illustrated through the localized monitoring of lactate in the human leg during and after exercise.

In vivo NMR evidence for moderate glucose accumulation in human skeletal muscle during hyperglycemia.

Although the absence of intracellular (IC) free glucose is direct evidence of glucose transport being the rate-limiting step for muscle glucose disposal at euglycemia, the scarcity of data in humans during hyperglycemia precludes any definitive conclusion. In the present study, 13C and 31P in vivo nuclear magnetic resonance (NMR) data from two separate groups of subjects were combined to measure IC free glucose in the human skeletal muscle. When these noninvasive tools were used with an infusion of [1-13C]glucose, a steady-state concentration of 1.2 +/- 0.2 mmol IC glucose/l IC water was observed at the end of a 2-h hyperglycemic clamp with somatostatin infusion, during which glycemia was maintained at approximately 22 mmol/l and insulinemia at approximately 5 mU/l. Despite this moderate glucose accumulation, the persistence of a large transmembrane glucose gradient suggests that the posttransport steps do not play a significant role in the control of muscle glucose disposal in these specific conditions, relevant to insulinopenic diabetic patients.

1H NMR determination of lactate 13C-enrichment in skeletal muscle: using a double quantum filter for the simultaneous editing of 13C-coupled and 13C-uncoupled methyl protons resonance.

1H NMR simultaneous editing of 13C-coupled and 13C-uncoupled methyl protons resonance, using the selection of double quantum coherences by a gradient pulse, was analyzed in vitro and demonstrated in situ on the hindlimb of an exercised rat model postmortem. In vitro calibration showed agreement with theoretical analysis. High-resolution NMR of muscle extract confirmed the accuracy of the lactate 13C-enrichment calculated using the in situ NMR data and the calibration factor obtained in vitro.