Phosphocreatine resynthesis during recovery in different muscles of the exercising leg by 31P‐MRS

To investigate the high‐energy phosphate metabolism by ³¹P‐nuclear magnetic resonance spectroscopy during off‐transition of exercise in different muscle groups, such as calf muscles and biceps femoris muscles, seven male long‐distance runners (LDR) and nine untrained males (UT) performed both submaximal constant and incremental exercises. The relative exercise intensity was set at 60% of the maximal work rate (60%Wmax) during both knee flexion and plantar flexion submaximal constant load exercises. The relative areas under the inorganic phosphate (P_i) and phosphocreatine (PCr) peaks were determined. During the 5‐min recovery following the 60%Wmax, the time constant for the PCr off‐kinetics was significantly faster in the plantar flexion (LDR: 17.3 ± 3.6 s, UT: 26.7 ± 6.7 s) than in the knee flexion (LDR: 29.7 ± 4.7 s, UT: 42.7 ± 2.8 s, P < 0.05). In addition, a significantly faster PCr off‐kinetics was observed in LDR than in UT for both exercises. The ratio of P_i to PCr (P_i/PCr) during exercise was significantly lower during the plantar flexion than during the knee flexion (P < 0.01). These findings indicated that the calf muscles had relatively higher potential for oxidative capacity than that of biceps femoris muscles with an association of training status.     

Effects of age on muscle energy metabolism and oxygenation in the forearm muscles

PURPOSE: The effects of aging on muscle metabolism and oxygenation have not yet been elucidated. We evaluated the effects of aging on energy metabolism and oxygenation in sedentary healthy subjects by simultaneously measuring 31P-magnetic resonance spectroscopy (MRS) and near-infrared spectroscopy (NIRS). METHODS: Nine young (28.1 +/- 5.0 yr) and nine older (61.4 +/- 4.6 yr) healthy subjects were studied. The 31P-MR spectrum was obtained every 15 s during and after hand gripping exercise. Intracellular pH (pHi) and PCr/(PCr+Pi) [PCr: phosphocreatine, Pi: inorganic phosphate] were calculated as an index of energy metabolism. The time constant of the PCr/(PCr+Pi) recovery (tau PCr) was calculated. With NIRS, we evaluated the recovery rates of oxygenated (RHbO2) and deoxygenated hemoglobin (RHb) during the initial 10 s of recovery. RESULTS: The PCr/(PCr+Pi) and pHi at rest and at completion of the exercise and tau PCr did not differ between young and older subjects. However, RHbO2 and RHb were significantly slower in older subjects than in young subjects. CONCLUSIONS: The results suggest that muscle energy metabolism in the forearm muscle was not affected by aging. The slower RHbO2 and RHb in older subjects suggested impaired O2 supply, which was probably due to impaired peripheral circulation caused by the process of aging.     

Phosphorus‐31 magnetic resonance spectroscopy of skeletal muscle in maternally inherited diabetes and deafness A3243G mitochondrial mutation carriers

Purpose

To investigate high‐energy phosphate metabolism in striated skeletal muscle of patients with Maternally Inherited Diabetes and Deafness (MIDD) syndrome.

Materials and Methods

In 11 patients with the MIDD mutation (six with diabetes mellitus [DM] and five non‐DM) and eight healthy subjects, phosphocreatine (PCr) and inorganic phosphate (Pi) in the vastus medialis muscle was measured immediately after exercise using ³¹P‐magnetic resonance spectroscopy (MRS). The half‐time of recovery (t1/2) of monoexponentially fitted (PCr+Pi)/PCr was calculated from spectra obtained every 4 seconds after cessation of exercise. A multiple linear regression model was used for statistical analysis.

Results

Patients with the MIDD mutation showed a significantly prolonged t1/2 (PCr+Pi)/PCr after exercise as compared to controls (13.6±3.0 vs. 8.7±1.3 sec, P = 0.01). No association between the presence of DM and t1/2 (PCr + Pi)/PCr was found (P = 0.382).

Conclusion

MIDD patients showed impaired mitochondrial oxidative phosphorylation in skeletal muscle shortly after exercise, irrespective of the presence of DM. J. Magn. Reson. Imaging 2009;29:127–131. © 2008 Wiley‐Liss, Inc.     

Comparison of measuring energy metabolism by different 31P‐magnetic resonance spectroscopy techniques in resting,ischemic, and exercising muscle

Alternate methods to quantify mitochondrial activity or function have been extensively used for studying insulin resistance and type 2 diabetes mellitus, namely saturation transfer and phosphocreatine (PCr) recovery. As these methods are in fact determining different parameters, this study aimed to compare saturation transfer results to PCr recovery measurements within the same group. Fifteen subjects underwent saturation transfer and ischemic exercise‐recovery experiments. PCr decrease during ischemia (Q), induced by cuff inflation, served as an additional measure of resting ATP (adenosine triphosphate) production. ATP synthetic rate (fATP) measured by saturation transfer (0.234 ± 0.043 mM/s) was greater than (Q = 0.0077 ± 0.0011 mM/s), but correlated well with Q (r = 0.63 P = 0.013). Parameters of PCr recovery correlated well with fATP (Q_max,lin: r = 0.71, P = 0.003, Q_max,ADP: r = 0.66, P = 0.007) and Q (Q_max,lin: r = 0.92, P = 0.000002, Q_max,ADP: r = 0.76, P = 0.001). In conclusion, although saturation transfer yields higher ATP synthetic rates than PCr decrease during ischemia, their significant correlation indicates that fATP can be used as a marker of mitochondrial activity. The finding that both Q and fATP correlate with PCr recovery kinetics suggests that skeletal muscle with greater maximal aerobic ATP synthetic rates is also metabolically more active at rest. Magn Reson Med, 2011. © 2011 Wiley‐Liss, Inc.     

Isokinetic eccentric exercise as a model to induce and reproduce pathophysiological alterations related to delayed onset muscle soreness

Physiological alterations following unaccustomed eccentric exercise in an isokinetic dynamometer of the right m. quadriceps until exhaustion were studied, in order to create a model in which the physiological responses to physiotherapy could be measured. In experiment I (exp. I), seven selected parameters were measured bilaterally in 7 healthy subjects at day 0 as a control value. Then after a standardized bout of eccentric exercise the same parameters were measured daily for the following 7 d (test values). The measured parameters were: the ratio of phosphocreatine to inorganic phosphate (PCr/P_i), the ratio of inorganic phosphate to adenosintriphosphate (Pi/ATP), the ratio of phosphocreatine to adenosintriphosphate (PCr/ATP) (all three ratios measured with ³¹P-nuclear magnetic resonance spectroscopy), dynamic muscle strength, plasma creatine kinase (CK), degree of pain and “muscle” blood flow rate (¹³³Xenon washout technique). This was repeated in experiment II (exp. II) 6–12 months later in order to study reproducbility. In experiment III (exp. III), the normal fluctuations over 8 d of the seven parameters were measured, without intervention with eccentric exercise in 6 other subjects. All subjects experienced pain, reaching a maximum 48 h after eccentric exercise in both exp. I and II. A systematic effect over time for CK (increasing 278% resp. 308%), muscle strength (decreasing more than 10%), PCr/Pi (decreasing 31% resp. 43%) and Pi/ATP (increasing 55% resp. 99%) was found in both exp. I and II (P<0.05), but not in exp. III. No significant difference was observed between exp. I and II for CK, blood-flow rate, concentric muscle strength, PCr/Pi, Pi/ATP and PCr/ATP. It is concluded that pathophysiological alterations in m. quadriceps following eccentric exercise can be induced and can be reproduced after an interval of 6 months. Thus, this model can be used to study the effects of physiotherapy.     

Metabolic changes in the rat brain after acute and chronic ethanol intoxication: A 31P NMR spectroscopy study

In this work, ³¹P phosphorus NMR (³¹P NMR) studies of the brain have been conducted in rats acutely and chronically intoxicated with ethanol. In both groups, changes in levels of high-energy phosphates were observed: increase of phosphocreatinine (PCr)/β AaTP and PCr/inorganic phosphate (P_i) in acute and long-term ethanol exposure, and decrease of P_iβ ATP after acute ethanol administration. These changes in high-energy phosphates, indicative of a reduction of adenosine triphosphate (ATP) and PCr consumption (PCr+ ADP + H⁺ ATP + Cr; ATP ADP + P_i), suggest a reduction of cerebral metabolism both in acute and chronic ethanol exposure. In addition, in the group of rats chronically intoxicated with ethanol, there were variations in phosphodiester peak intensities (decrease of phosphomonoester (PME)/phosphodiester (PDE), increase of PDE/β ATP), suggesting increased breakdown of membrane phospholipids. These changes could provide a metabolic explanation for the development of cerebral atrophy in chronic alcoholism.     

Comparisons of ATP turnover in human muscle during ischemic and aerobic exercise using 31P magnetic resonance spectroscopy

To investigate human muscle bioenergetics quantitatively in vivo, we used ³¹P magnetic resonance spectroscopy to study the flexor digitorum superficialis of four adult males during dynamic ischemic and aerobic exercise at 0.50–1.00 W and during recovery from aerobic exercise. During exercise, changes in pH and [PCr] were larger at higher power, but in aerobic exercise neither end-exercise [ADP] nor the initial postexercise PCr resynthesis rate altered with power. In ischemic exercise we estimated total ATP synthesis from the rates of PCr depletion and glycogenolysis (inferred using an analysis of proton buffering); this was linear with power output. In aerobic exercise, again we estimated ATP synthesis rates due to phosphocreatine hydrolysis and glycogenolysis (incorporating a correction for proton efflux) and also estimated oxidative ATP synthesis by difference, using the total ATP turnover rate established during ischemic exercise. We conclude that in early exercise oxidative ATP synthesis was small, increasing by the end of exercise to a value close (as predicted) to the initial postexercise rate of PCr resynthesis. Furthermore, a plausible estimate of proton efflux during aerobic exercise can be inferred from the pH-dependence of proton efflux in recovery.     

31P NMR saturation transfer study of the creatine kinase reaction in human skeletal muscle at rest and during exercise

The creatine kinase reaction has been studied by ³¹P NMR in exercising human calf muscle. Quantitative analysis of high energy phosphates and saturation transfer study of the creatine kinase flux in the direction of ATP synthesis (V_for) were performed at rest and during exercise. As expected, exercise induced a [PCr] decrease (from 28.5 ± 0.9 to 21.9 ± 1.5 mM, P < 0.01) matched by a P₁, increase (from 4.5 ± 0.2 to 8.9 ± 1.8 mM,P = 0.06). pH_i and [ATP] remained unchanged. V_for did not change from rest (12.4 ± 0.9 mM s^?1) to moderate exercise and decreased at the highest exercise level (8.4 ± 1.4 mM s^?1, P = 0.006). This observation differs from the prediction of the creatine kinase rate equation, showing an increase in the flux with exercise intensity. Computations suggest that this discrepancy arises from metabolite compartmentalization and/or from the reaction kinetics of a dead end complex stabilized by planar anions.     

Exertional heatstroke and muscle metabolism: an in vivo 31P-MRS study.

An impairment of muscle energy metabolism has been suggested as a predisposing factor for, as well as a consequence of exertional heatstroke (EHS). Thirteen young men were investigated 6 months after a well-documented EHS using 31Phosphorus Magnetic Resonance Spectroscopy (31P-MRS). The relative concentrations of ATP, phosphocreatine (PCr), inorganic phosphate (Pi), phosphomonoesters (PME), and the intracellular pH (pHi) were determined at rest, during a graded standardized exercise protocol (360 active plantar flexions) and during recovery. Also the leg tissue blood flow was determined by venous occlusion plethysmography during the MRS procedure. Sixteen age-matched healthy male volunteers served as control group. In resting muscle, there were no significant differences between the groups as regards pHi, Pi/PCr, and ATP/PCr+Pi+PME ratios. During steady state exercise conditions, effective power outputs were similar for both groups at each level of exercise: 20, 35, and 50% of maximal voluntary contraction (MVC) of the calf muscle. No significant differences were shown between the two groups in Pi/PCr, pHi, or changes of leg blood flow at each level of exercise. At 50% MVC, Pi/PCr was 0.48 +/- 0.08 vs 0.47 +/- 0.05 (P = 0.96), pHi was 6.94 +/- 0.03 vs 6.99 +/- 0.02, respectively (P = 0.13). Finally, the rate of PCr resynthesis during recovery was not significantly different between the two groups: t1/2 PCr = 0.58 +/- 0.07 vs 0.50 +/- 0.05 min, respectively (P = 0.35). Therefore, no evidence of an impairment of muscle energy metabolism was shown in the EHS group during a standardized submaximal exercise using 31P-MRS performed 6 months after an EHS.     

Factors affecting the rate of phosphocreatine resynthesis following intense exercise

Within the skeletal muscle cell at the onset of muscular contraction, phosphocreatine (PCr) represents the most immediate reserve for the rephosphorylation of adenosine triphosphate (ATP). As a result, its concentration can be reduced to less than 30% of resting levels during intense exercise. As a fall in the level of PCr appears to adversely affect muscle contraction, and therefore power output in a subsequent bout, maximising the rate of PCr resynthesis during a brief recovery period will be of benefit to an athlete involved in activities which demand intermittent exercise. Although this resynthesis process simply involves the rephosphorylation of creatine by aerobically produced ATP (with the release of protons), it has both a fast and slow component, each proceeding at a rate that is controlled by different components of the creatine kinase equilibrium. The initial fast phase appears to proceed at a rate independent of muscle pH. Instead, its rate appears to be controlled by adenosine diphosphate (ADP) levels; either directly through its free cytosolic concentration, or indirectly, through its effect on the free energy of ATP hydrolysis. Once this fast phase of recovery is complete, there is a secondary slower phase that appears almost certainly rate-dependent on the return of the muscle cell to homeostatic intracellular pH. Given the importance of oxidative phosphorylation in this resynthesis process, those individuals with an elevated aerobic power should be able to resynthesise PCr at a more rapid rate than their sedentary counterparts. However, results from studies that have used phosphorus nuclear magnetic resonance ((31)P-NMR) spectroscopy, have been somewhat inconsistent with respect to the relationship between aerobic power and PCr recovery following intense exercise. Because of the methodological constraints that appear to have limited a number of these studies, further research in this area is warranted.     

Tumor 31P NMR pH measurements in vivo: A comparison of inorganic phosphate and intracellular 2-deoxyglucose-6-phosphate as pHnmr indicators in murine radiation-induced fibrosarcoma-1

Uncertainty regarding the intracellular/extracellular distribution of inorganic phosphate (P_i) in tumors has raised concerns that pH calculated from the tumor P_i chemical shift may not accurately represent the intracellular pH (pH_in). This issue was addressed in subcutaneously transplanted murine radiation induced fibrosarcoma-1 by directly comparing pH measured via P_i with pH measured via the in situ generated intracellular xenometabolite 2-deoxyglucose-6-phosphate (2DG6P). In 131 comparative measurements employing eight tumor-bearing mice under both control and hyperglycemic conditions (the latter to extend the range of tumor pH examined), the pH as derived from either 2DG6P or P_i showed only a small, but statistically significant, difference (0.07 ± 0.11 SD; P = 0.0001). Scatter in the comparative analysis over the pH range examined (ca. 5.5-7.5) was not uniform. Above pH 6.6, 2DG6P indicated a pH lower than that of P_i by 0.088 ± 0.105 SD (n = 107, P = 0.0001); below pH 6.6, 2DG6P indicated a pH essentially identical to and not statistically different from that of P_i (mean difference 0.003 ± 0.128 SD (n = 24, P = 0.92)). Evidence is presented in support of this differential arising from a systematic measurement error due to peak overlap between 2DG6P and endogenous phosphomonoester species. These results support the use of P_i as a tumor ³¹P NMR pH_in indicator, at least in RIF-1 tumors under control and hyperglycemic conditions.     

Transverse relaxation and magnetization transfer in skeletal muscle: Effect of pH

Exercise increases the intracellular T₂ (T_2,i) of contracting muscles. The mechanism(s) for the T_2,i increase have not been fully described, and may include increased intracellular free water and acidification. These changes may alter chemical exchange processes between intracellular free water and proteins. In this study, the hypotheses were tested that (a) pH changes T_2,i by affecting the rate of magnetization transfer (MT) between free intracellular water and intracellular proteins, and (b) the magnitude of the T_2,i effect depends on acquisition mode (localized or nonlocalized) and echo spacing. Frog gastrocnemius muscles were excised and their intracellular pH was either kept at physiological pH (7.0) or modified to model exercising muscle (pH 6.5). The intracellular transverse relaxation rate (R_2,i = 1/T_2,i) always decreased in the acidic muscles, but the changes were greater when measured using more rapid refocusing rates. The MT rate from the macromolecular proton pool to the free water proton pool, its reverse rate, and the spin‐lattice relaxation rate of water decreased in acidic muscles. It is concluded that intracellular acidification alters the R_2,i of muscle water in a refocusing rate‐dependent manner, and that the R_2,i changes are correlated with changes in the MT rate between macromolecules and free intracellular water. Magn Reson Med, 2009. © 2008 Wiley‐Liss, Inc.     

Skeletal muscle water and electrolytes following prolonged dehydrating exercise

We studied if dehydrating exercise would reduce muscle water (H₂O_muscle) and affect muscle electrolyte concentrations. Vastus lateralis muscle biopsies were collected prior, immediately after, and 1 and 4 h after prolonged dehydrating exercise (150 min at 33 ± 1 °C, 25% ± 2% humidity) on nine endurance‐trained cyclists (VO_2max = 54.4 ± 1.05 mL/kg/min). Plasma volume (PV) changes and fluid shifts between compartments (Cl^? method) were measured. Exercise dehydrated subjects 4.7% ± 0.3% of body mass by losing 2.75 ± 0.15 L of water and reducing PV 18.4% ± 1% below pre‐exercise values (P < 0.05). Right after exercise H₂O_muscle remained at pre‐exercise values (i.e., 398 ± 6 mL/100 g dw muscle^?1) but declined 13% ± 2% (342 ± 12 mL/100 g dw muscle^?1; P < 0.05) after 1 h of supine rest. At that time, PV recovered toward pre‐exercise levels. The Cl^? method corroborated the shift of fluid between extracellular and intracellular compartments. After 4 h of recovery, PV returned to pre‐exercise values; however, H₂O_muscle remained reduced at the same level. Muscle Na⁺ and K⁺ increased (P < 0.05) in response to the H₂O_muscle reductions. Our findings suggest that active skeletal muscle does not show a net loss of H₂O during prolonged dehydrating exercise. However, during the first hour of recovery H₂O_muscle decreases seemly to restore PV and thus cardiovascular stability.     

Comparing localized and nonlocalized dynamic 31P magnetic resonance spectroscopy in exercising muscle at 7T

By improving spatial and anatomical specificity, localized spectroscopy can enhance the power and accuracy of the quantitative analysis of cellular metabolism and bioenergetics. Localized and nonlocalized dynamic ³¹P magnetic resonance spectroscopy using a surface coil was compared during aerobic exercise and recovery of human calf muscle. For localization, a short echo time single‐voxel magnetic resonance spectroscopy sequence with adiabatic refocusing (semi‐LASER) was applied, enabling the quantification of phosphocreatine, inorganic phosphate, and pH value in a single muscle (medial gastrocnemius) in single shots (T_R = 6 s). All measurements were performed in a 7 T whole body scanner with a nonmagnetic ergometer. From a series of equal exercise bouts we conclude that: (a) with localization, measured phosphocreatine declines in exercise to a lower value (79 ± 7% cf. 53 ± 10%, P = 0.002), (b) phosphocreatine recovery shows shorter half time (t_1/2 = 34 ± 7 s cf. t_1/2 = 42 ± 7 s, nonsignificant) and initial postexercise phosphocreatine resynthesis rate is significantly higher (32 ± 5 mM/min cf. 17 ± 4 mM/min, P = 0.001) and (c) in contrast to nonlocalized ³¹P magnetic resonance spectroscopy, no splitting of the inorganic phosphate peak is observed during exercise or recovery, just an increase in line width during exercise. This confirms the absence of contaminating signals originating from weaker‐exercising muscle, while an observed inorganic phosphate line broadening most probably reflects variations across fibers in a single muscle. Magn Reson Med, 2012. © 2012 Wiley Periodicals, Inc.     

Impact of salbutamol on muscle metabolism assessed by 31P NMR spectroscopy

The potential ergogenic effects of oral salbutamol intake were demonstrated for decades but the underlying mechanisms remain to elucidate. We hypothesized that improved exercise performance after acute oral salbutamol administration is associated with changes in muscle metabolism. Twelve healthy, nonasthmatic, moderately trained, male subjects were recruited to compare in a double‐blind crossover randomized study, an oral dose of salbutamol (4 mg) and a placebo. After treatment administration, subjects performed repetitive plantar flexions to exhaustion in a 3T magnet. Continuous ³¹P nuclear magnetic resonance spectroscopy assessment of the calf muscles was performed at rest, during exercise, and during recovery. No significant difference between treatments was detected in metabolite concentration at rest (P > 0.05). Creatine phosphate and inorganic phosphate changes during and immediately after exercise were similar between treatments (P > 0.05). Intramuscular pH (pHi) was significantly higher at rest, at submaximal exercise but not at exhaustion with salbutamol (pHi at 50% of exercise duration, 6.8 ± 0.1/6.9 ± 0.1 for placebo and salbutamol, respectively, P < 0.05). The maximal power (28 ± 7 W/23 ± 7 W; P = 0.001) and total work (1702 ± 442 J/1381 ± 432 J; P = 0.003) performed during plantar flexions were significantly increased with salbutamol. Salbutamol induced significant improvement in calf muscle endurance with similar metabolic responses during exercise, except slight differences in pHi. Other mechanisms than changes in muscle metabolism may be responsible for the ergogenic effect of salbutamol administration.     

ATP production and mechanical work in exercising skeletal muscle: A theoretical analysis applied to 31P magnetic resonance spectroscopic studies of dialyzed uremic patients

³¹P magnetic resonance spectroscopy (³¹P MRS) can yield much information about bioenergetics in skeletal muscle. During mixed aerobic/glycolytic exercise, changes in phos-phocreatine (PCr) concentration and pH may be abnormal because of reduced muscle mass or reduced efficiency (which the authors combine here as “effective muscle mass”) or because of reduced oxidative capacity. The authors show how these can be distinguished by calculating the nonoxidative and oxidative costs of mechanical work, and also of work per unit of effective muscle mass (measured using the initial rate of ATP turnover). These quantities are substantially time-independent during incremental exercise, and so can be used to compare exercise studies of differing duration. The authors illustrate this analysis by showing that in dialyzed patients with chronic renal failure, the substantial exercise abnormalities seen by ³¹P MRS are due mainly to a decrease in effective muscle mass, which outweighs the oxidative defect implied by the abnormal PCr recovery kinetics.     

Energetics of 3.5 s neural activation in humans: A 31P MR spectroscopy study

No direct information on brain energetics and energy-related compounds in the first seconds of physiological activation has been reported to date. In this study visual cortex high energy phosphate changes were monitored in 11 normal subjects during 3.5 s activation and the following 23.5 s by a simple ³¹P magnetic resonance spectroscopic method. An intraactivation decrease of phosphocreatine (PCr) was observed in all subjects, with changes in pH in three, one of them also presenting a change in adenosine triphosphate (ATP). In the subgroup of eight subjects without changes in pH, the mean rate of mean PCr decrease (D_PCr) was 7.24 ± 0.78 %/s, and the postactivation mean rate of mean PCr recovery was <1/2 D_PCr. Short phasic neural activity requires a large amount of energy, i.e., at least three times basal consumption, in agreement with theoretical calculations. Additional energy demands in the visual cortex are several times those measured by positron emission tomography during prolonged stimulation studies, implying that mean energy requirements decrease with increases in duration of stimulation. During short activation, the vascular responses as detected by brainmapping techniques (BMT) are preceded by an important reduction of the intracellular high-energy phosphate content, which returns to resting values during an interval that corresponds to the poststimulation return of BMT signals to baseline.     

Evaluations of cooling exercised muscle with MR imaging and 31P MR spectroscopy

PURPOSE: To investigate the effects of cooling human skeletal muscle after strenuous exercise using 31P MR spectroscopy and MR imaging. METHODS: 14 male subjects (mean age +/- SD, 23.8 +/- 2.3 yr) were randomly assigned to the normal (N = 7) or the cooling group (N = 7). All subjects performed the ankle plantar flexion exercise (12 repetitions, 5 sets). Localized 31P-spectra were collected from the medial gastrocnemius before and after exercise (immediately, 30, 60 min, 24, 48, 96, and 168 h) to determine the ratio of inorganic phosphate to phosphocreatine (Pi/PCr) and intracellular pH. Transaxial T2-weighted MR images of the medial gastrocnemius were obtained to calculate T2 relaxation time (T2), indicative of intramuscular water level, before and after exercise (24, 48, 96, and 168 h). In addition, the muscle soreness level was assessed at the same time as 31P-spectra measurements. Fifteen-minute cold-water immersion was administered to the cooling group after exercise and initial postexercise measurements. RESULTS: The control group showed significantly increased T2 from rest at 48 h after exercise (P < 0.05), but the cooling group showed no significant change in T2 throughout this study. Both groups showed a significantly decreased intracellular pH immediately after exercise (P < 0.05). After that, the cooling group showed a significantly greater value than the value at rest or the control group at 60 min after exercise (P < 0.05). For the Pi/PCr, no significant change was observed in both groups throughout this study. The muscle soreness level significantly increased immediately and at 24-48 h after exercise in both groups (P < 0.05). CONCLUSION: The findings of this study suggest that cooling causes an increase in intracellular pH and prevents the delayed muscle edema.     

Modeling in vivo recovery of intracellular pH in muscle to provide a novel index of proton handling: application to the diagnosis of mitochondrial myopathy.

Post-exercise recovery of intracellular pH (pH(i)) assessed using phosphorus magnetic resonance spectroscopy has not been previously evaluated in its entirety due to its complex time-course and missing data points resulting from a transient loss of inorganic phosphate signal. By considering the transition from exercise to recovery as a step function input, pH(i) recovery was modeled based on the creatine-kinase equilibrium, and the entire pH(i) recovery was characterized by calculating the time required for pH(i) recovery (t(pHrec)). Applying this methodology, normal subjects showed a strong linear correlation between phosphocreatine (PCr) half-time and t(pHrec) (r = 0.90, P < 0.001). In mitochondrial myopathy (MM) patients with weakness in the limb examined, 9/10 had faster pH(i) recovery relative to PCr recovery; wide normal ranges from a control group which included deconditioned subjects resulted in 7 of those 10 patients having otherwise normal recovery indices. Therefore, modeling pH(i) recovery allows characterization of the entire pH(i) recovery and detects altered proton handling in MM patients, including those with otherwise normal recovery indices.     

Homocarnosine and the measurement of neuronal pH in patients with epilepsy

Homocarnosine is a dipeptide of gamma-aminobutyric acid (GABA) and histidine found uniquely in the brain, most likely in a subclass of GABAergic neurons. By comparison of spectra from the occipital lobe of patients receiving a homocarnosine elevation drug to normal subjects we have assigned two elevated resonances in the short TE ¹H MRS spectrum to homocarnosine. These resonances are partially resolved at 7.05 and 8.02 ppm in a short TE spectrum at 2.1 T when macromolecule resonances are removed by subtraction of a spectrum in which the metabolite resonances are nulled by inversion recovery. The chemical shift of both of these resonances is sensitive to pH_i. By comparison with a titration curve the pH_i was calculated from the downfield resonance to be 7.06 in the patient group which is similar to values reported using the P_i resonance. Based on the in vivo results and theoretical considerations the potential sensitivity for using nonelevated homocarnosine to measure pH is similar to that of P_i under physiological conditions.