Origin of the 31 P MR signal at 5.3 ppm in patients with critical limb ischemia.

An unknown intense signal (Pun ) with a mean chemical shift of 5.3 ppm was observed in 31 P MR spectra from the calf muscles of patients with the diabetic foot syndrome. The aim of the study was to identify the origin of this signal and its potential as a biomarker of muscle injury. Calf muscles of 68 diabetic patients (66.3 ± 8.6 years; body mass index = 28.2 ± 4.3 kg/m2 ) and 12 age-matched healthy controls were examined by (dynamic) 31 P MRS (3 T system, 31 P/1 H coil). Phantoms (glucose-1-phosphate, Pi and PCr) were measured at pH values of 7.05 and 7.51. At rest, Pun signals with intensities higher than 50% of the Pi intensity were observed in 10 of the 68 examined diabetic subjects. We tested two hypothetical origins of the Pun signal: (1) phosphorus from phosphoesters and (2) phosphorus from extra- and intracellular alkaline phosphate pools. 2,3-diphosphoglycerate and glucose-1-phosphate are the only phosphoesters with signals in the chemical shift region close to 5.3 ppm. Both compounds can be excluded: 2,3-diphosphoglycerate due to the missing second signal component at 6.31 ppm; glucose-1-phosphate because its chemical shifts are about 0.2 ppm downfield from the Pi signal (4.9 ppm). If the Pun signal is from phosphate, it represents a pH value of 7.54 ± 0.05. Therefore, it could correspond to signals of Pi in mitochondria. However, patients with critical limb ischemia have rather few mitochondria and so the Pun signal probably originates from interstitia. Our data suggest that the increased Pun signal observed in patients with the diabetic foot syndrome is a biomarker of severe muscular damage.

Different acute effects of fructose and glucose administration on hepatic fat content.

BACKGROUND: Diets rich in fat and added sugars (especially fructose) play an important role in the pathogenesis of nonalcoholic liver disease (NAFLD), but there is only limited information on the acute effects of these nutrients on hepatic fat content (HFC). OBJECTIVES: We therefore explored how the administration of high-fat load, glucose, fructose, and combinations thereof affects HFC measured in vivo using proton magnetic resonance spectroscopy (1H-MRS) in healthy subjects. METHODS: Ten healthy nonsteatotic male volunteers (age 38.5 ± 9.6 y, body mass index [BMI, kg/m2] 26.9 ± 2.7) underwent, in random order, 6 experiments, each lasting 8 h, that included: 1) fasting; 2) a high-fat load (150 g of fat [dairy cream] at time 0); 3) glucose (3 doses of 50 g at 0, 2, and 4 h); 4) a high-fat load with glucose; 5) fructose (3 doses of 50 g at 0, 2, and 4 h); and 6) a high-fat load with fructose. HFC was measured using 1H-MRS prior to test meal administration (before time 0) and at 3 and 6 h. Plasma concentrations of triglycerides, nonesterified fatty acids, glucose, and insulin were monitored throughout each experiment. RESULTS: HFC increased to 119 ± 19% (P < 0.05) and 117 ± 17% (P < 0.01) of baseline when subjects consumed a high-fat load alone or a high-fat load with fructose, respectively, but was not affected when glucose was coadministered with a high-fat load. HFC was not affected when subjects had fasted or had consumed repeated doses of fructose. When subjects were administered 3 doses of glucose, HFC dropped to 85 ± 13% (P < 0.05) of baseline. CONCLUSIONS: Our results demonstrate that fructose and glucose have a different immediate impact on HFC in humans in vivo. Clinical trial registry: The study was registered at clinicaltrials.gov and obtained clinicaltrials.gov identifier: NCT03680248.

MR compatible ergometers for dynamic 31P MRS.

Magnetic Resonance (MR) compatible ergometers are specialized ergometers used inside the MR scanners for the characterization of tissue metabolism changes during physical stress. They are most commonly used for dynamic phosphorous magnetic resonance spectroscopy (31P MRS), but can also be used for lactate production measurements, perfusion studies using arterial spin labelling or muscle oxygenation measurements by blood oxygen dependent contrast sequences. We will primarily discuss the importance of ergometers in the context of dynamic 31P MRS. Dynamic 31P MRS can monitor muscle fatigue and energy reserve during muscle contractions as well as the dynamics of recuperation of skeletal muscle tissue during the following recovery through signal changes of phosphocreatine (PCr), inorganic phosphate and adenosine triphosphate (ATP). Based on the measured data it is possible to calculate intracellular pH, metabolic flux of ATP through creatine-kinase reaction, anaerobic glycolysis and oxidative phosphorylation and other metabolic parameters as mitochondrial capacity. This review primarily focuses on describing various technical designs of MR compatible ergometers for dynamic 31P MRS that must be constructed with respect to the presence of magnetic field. It is also expected that the construction of ergometers will be easy for the handling and well accepted by examined subjects.

31P-MR spectroscopy in patients with mild and serious lower limb ischemia.

BACKGROUND: 31P-MR spectroscopy is a technique for undertaking a comprehensive evaluation of muscle metabolism. The goal of this study was to compare patients with mild and severe lower limb ischemia measured by 31P-MR spectroscopy at rest and during exercise. METHODS: Sixteen non-diabetic mild peripheral arterial occlusive disease (PAOD) patients, 23 diabetic PAOD patients with severe ischemia and 19 healthy controls were examined by rest and dynamic 31P-MR spectroscopy with a 3T MR system equipped with an MR-compatible home-made pedal ergometer. Signal intensity ratios of phosphorous metabolites to the sum of all 31P intensities (Ptot) and pH were obtained at rest. The PCr drop (ΔPCr), time recovery constant of PCr (τPCr), pH at the end of the exercise (pHend), and mitochondrial capacity (Qmax) were calculated from dynamic MR spectra. RESULTS: Diabetic PAOD patients with severe ischemia differed from controls in both rest (PCr/Pi, ßATP/Ptot, pH) and dynamic (Qmax, pHend, τPCr) parameters. PAOD patients with mild ischemia differed from controls only in Qmax and pHend. Rest parameters of the nondiabetic PAOD patients did not differ from control values excluding rest pH which was higher in both patient groups. CONCLUSIONS: A combination of rest and dynamic 31P-MR spectroscopy can distinguish among all three groups of subjects. On the other hand, examination at rest is sufficient for differentiation between patient groups and verification of severe ischemia.

[Diabetic foot syndrome: importance of calf muscles MR spectroscopy in the assessment of limb ischemia and effect of revascularization]. / Syndrom diabetické nohy: význam MR spektroskopie lýtkových svalu pro hodnocení koncetinové ischemie a efektu revaskularizace.

AIM: The standard method for assessment of effect of revascularization in patients with diabetic foot (DF) and critical limb ischemia (CLI) is transcutaneous oxygen pressure (TcPO2). Phosphorus magnetic resonance spectroscopy (31P MRS) enables to evaluate oxidative muscle metabolism that could be impaired in patients with diabetes and its complications. The aim of our study was to compare MRS of calf muscle between patients with DF and CLI and healthy controls and to evaluate the contribution of MRS in the assessment of the effect of revascularization. METHODS: Thirty-four diabetic patients with DF and CLI treated either by autologous cell therapy (ACT; 15 patients) or percutaneous transluminal angioplasty (PTA; 12 patients) in our foot clinic during 2013-2016 and 19 healthy controls were included into the study. TcPO2 measurement was used as a standard method of non-invasive evaluation of limb ischemia. MRS examinations were performed using the whole-body 3T MR system 1 day before and 3 months after the procedure. Subjects were examined in a supine position with the coil fixed under the m. gastrocnemius. MRS parameters were obtained at rest and during the exercise period. Rest MRS parameters of oxidative muscle metabolism such as phosphocreatine (PCr), inorganic phosphate (Pi), phosphodiesters (PDE), adenosine triphosphate (ATP), dynamic MRS parameters such as recovery constant PCr (τPCr) and mitochondrial capacity (Qmax), and pH were compared between patients and healthy controls, and also before and 3 months after revascularization. RESULTS: Patients with CLI had significantly lower PCr/Pi (p < 0.001), significantly higher Pi and pH (both p < 0.01), significantly lower Qmax and prolonged τPCr (both p < 0.001) in comparison with healthy controls. We observed a significant improvement in TcPO2 at 3 months after revascularization (from 26.4 ± 11.7 to 39.7 ± 17.7 mm Hg, p < 0.005). However, the rest MRS parameters did not change significantly after revascularization. In individual cases we observed improvement of dynamic MRS parameters. There was no correlation between MRS parameters and TcPO2 values. CONCLUSION: Results of our study show impaired oxidative metabolism of calf muscles in patients with CLI in comparison with healthy controls. We observed an improvement in dynamic MRS parameters in individual cases; this finding should be verified in a large number of patients during longer follow-up.Key words: autologous cell therapy - critical limb ischemia - diabetic foot - MR spectroscopy.

Dynamic 31P MR spectroscopy of plantar flexion: influence of ergometer design, magnetic field strength (3 and 7 T), and RF-coil design.

PURPOSE: Dynamic phosphorus magnetic resonance spectroscopy ((31)P MRS) during and after acute exercise enables the noninvasive in vivo determination of the mitochondrial capacity of skeletal muscle. Nevertheless, the lack of standardization in experimental setups leads to significant variations in published values of maximal aerobic capacity, even in the population of healthy volunteers. Thus, in this study, we aimed to assess the impact of the ergometer type (pneumatic and mechanical resistance construction), radiofrequency (RF)-coil diameter, and different magnetic field strengths (3 and 7 T) on the metabolic parameters measured by dynamic (31)P MRS during a plantar flexion isotonic exercise protocol within the same group of healthy volunteers. METHODS: Dynamic (31)P MRS measurements of the calf muscle in 11 volunteers (mean age, 36 ± 13 yrs; mean BMI, 23.5 ± 2.5 kg/m(2)), on a 3 T MR system with a custom-made mechanical ergometer in the first research laboratory (RL1) and on 3 and 7 T MR systems equipped with a commercial pneumatic ergometer in the second research laboratory (RL2), were performed at three different workloads. RF-coils differed slightly between the sites and MR systems used. The repeatability of the experimental protocol was tested in every setup. The basal concentrations of phosphocreatine (PCr), exercise-induced depletion of PCr (ΔPCr), initial PCr resynthesis rate (VPCr), and mitochondrial capacity (Qmax) were calculated and compared between the research sites and field strengths. RESULTS: High repeatability of the measurement protocol was found in every experimental setup. No significant differences at any workload were found in these metabolic parameters assessed at different magnetic field strengths (3 T vs 7 T), using the same ergometer (in RL2) and a similar RF-coil. In the inter-research laboratory comparison at the same field strength (3 T), but with using different ergometers and RF-coils, differences were found in the concentration of PCr measured at rest and in the drop in PCr signal intensity. These differences translated into difference in the value of mitochondrial capacity at a workload of 15% of maximal voluntary contraction (MVC) force (0.45 ± 0.16 mM/s vs 0.31 ± 0.08 mM/s, in the RL1 and RL2, respectively). CONCLUSIONS: Metabolic parameters measured during exercise challenge by dynamic (31)P MRS do not depend upon the magnetic field strength used. For multicenter studies with different ergometers, it is important to set the same workload, measurement, and evaluation protocols, especially when the effects of very mild exercise (15% MVC) are to be compared. However, a higher workload (24% MVC) decreases the influence of imperfections and intersite differences for the assessed value of maximal mitochondrial capacity.