Mechanisms of31P relaxation in phosphorus metabolites

Measurements have been made of the longitudinal relaxation timeT ₁ of³¹P for the individual resonances of the metabolites AMP, ADP, ATP, P_ir and PCr (phosphocreatine) in H₂O and D₂O solutions from 5 to 60°C at various concentrations and at frequencies of 40 MHz (2.3 T) and 120 MHz (7 T). The contributions of dipolar, chemical shift anisotropy, and spin-rotation mechanisms have been separated, and activation parameters of the underlying molecular reorientations have been determined.     

Use of spin-traps during warm ischemia-reperfusion in rat liver: comparative effect on energetic metabolism studied using31P nuclear magnetic resonance

Detection of free radicals by electron spin resonance (ESR) proves the involvement of reactive oxygen species (ROS) in reperfused organ injuries. Spin-traps are known to ameliorate hemodynamic parameters in an isolated postischemic heart. The effects of 5 mmol/L DMPO (5,5-dimethyl-1-pyrroline-N-oxide) or DEPMPO (5-(diethylphosphoryl)-5-methyl-1-pyrrolineN-oxide) on intracellular pH (pH_in) and ATP level were evaluated by³¹P nuclear magnetic resonance on isolated rat liver submitted to 1 hour of warm ischemia and reperfusion. At the end of the reperfusion period, during which pH_in recovered to its initial value (7.16±0.03) in all groups, the ATP recovery level (expressed in percentage of initial value) was similar in controls and DEPMPO (60%±5%,n=6 and 54%±4%,n=6, respectively), but only 37%±1% in DMPO-treated livers (n=6) (p<0.05 versus controls andp<0.05 versus DEPMPO). Oxidative phosphorylation was not affected by an addition of nitrones on isolated mitochondria extracted from livers not submitted to ischemia-reperfusion. In contrast, mitochondria extracted at the end of the ischemia-reperfusion showed an impairment in the phosphorylation parameters, particularly in the presence of DMPO. Mass spectrum of ischemic liver perchloric acid extracts evidenced probable catabolites in treated groups. The differences in the effect of the two nitrones on energetic metabolism may be explained by the production of deleterious catabolites by DMPO as compared to DEPMPO. Even though a specific radical scavenging effect could be operative in the liver, our results indicate that catabolic effects were predominant. The absence of deleterious effects of DEPMPO in contrast to DMPO on the liver energetic metabolism was evidenced, allowing the use of DEPMPO for ESR detection.     

Combined 1H and 31P MR spectroscopic imaging: impaired energy metabolism in severe carotid stenosis and changes upon treatment

OBJECT: To evaluate if combined (1)H and (31)P MR spectroscopic imaging (MRSI) before and after treatment of severe internal carotid artery (ICA) stenosis detects significant changes in energy metabolism in the basal ganglia of both hemispheres. MATERIALS AND METHODS: A group of 14 patients with high-grade ICA stenosis and 11 healthy control subjects were examined with 2D (1)H MRSI and 3D (31)P MRSI at 3 T before and after treatment of severe ICA stenosis. Spectroscopic data were processed with LCModel and jMRUI software. Changes of the phosphorylated metabolites, pH, N-acetyl-acetate, creatine and choline-containing compounds prior/post intervention were analyzed and patients' data were compared with that of control subjects. RESULTS: Untreated patients had significantly higher Adenosindiphosphate (ADP) in basal ganglia ipsi- and contralateral to the side of ACI stenosis compared to controls. After treatment, ADP of both hemispheres significantly decreased by approximately 20% compared to the pre-treatment values. Further, significant decreases of phosphorylated metabolites prior/post intervention were found for patients compared to controls. CONCLUSION: This spectroscopic study reveals that unilateral high-grade ICA stenosis has an effect on cerebral high-energy metabolism of both hemispheres, which is at least partially reversible after treatment. Therefore the restoration of blood flow in high-grade ICA stenosis recovers the impaired energy balance of the brain.     

Functional and metabolic evaluation of the hypertrophied heart using MRI and31P-MRS

4. Conclusions Diastolic LV function and myocardial HEP metabolism are impaired only when LVH is caused by permanent pressure or volume overload, and not by a temporary increase in cardiac workload during part of the day as in elite athletes. Therefore, training-induced and pressure/volume-overload-induced LVH seem to represent different phenotypes of LVH, possibly related to genetic reprogramming which only occurs during permanent cardiac overload [17]. Moreover, there is an association between impaired LV diastolic function and altered myocardial HEP metabolism in patients with hypertension and in patients with aortic valve disease. Finally we did not find a correlation between myocardial HEP metabolism and LV mass in any of the groups studied. The latter indicates that LVH should be regarded as an epiphenomenon to cardiac overload, and not as a primary factor causing abnormal HEP metabolism.     

Investigation of multidrug resistance in cultured human renal cell carcinoma cells by 31P-NMR spectroscopy and treatment survival assays

KTCTL-26 and KTCTL-2 are renal cell carcinoma (RCC) lines with high and lowexpression of P-170 glycoprotein, respectively. Inherent differences between the two cell lines in terms of phosphate metabolites and growth characteristics in culture were examined for possible association with multidrug resistance (MDR). Differences in response to drug treatment were investigated for 40 h incubations with various doses of vinblastine (VBL) alone or as cotreatments with various concentrations of the calcium antagonist diltiazem (DIL) and/or interferon–α (IFN-α). Treatment effects were quantitated using the MTT survival assay and ³¹P magnetic resonance spectroscopy (MRS) to determine phosphate metabolite profiles in intact cells. KTCTL-2 and KTCTL-26 cells exhibited significant inherent differences in phosphocholine, glycerophosphocholine, glycerophosphoethanolamine, and phosphocreatine levels. KTCTL-26 cells were more sensitive than KTCTL-2 to 0.011μM VBL alone (87% vs. 102% survival) or to 0.011μM BL + 10μM DIL (55% vs. 80% survival). The latter treatment resulted in a significant decrease in the ratio of phosphocholine to glycerophosphocholine in KTCTL-26 cells but no significant changes in phosphate metabolites in KTCTL-2 cells. Metabolomic ³¹P MRS detects different metabolite profiles for RCC cell lines with different MDR phenotypes and may be useful for noninvasive characterization of tumors in a clinical setting.This revised version was published online in August 2005 with a corrected sequence of authors.     

The magnitude of signal errors introduced by ISIS in quantitative31P MRS

It is well known that the quality of a quantitative³¹P MRS measurement relies largely on the performance of the volume selection method, and that image selected in vivo spectroscopy (ISIS) suffers from contaminating signal caused mostly by Tl smearing. However, these signal errors and their magnitude are seldom addressed in clinical studies. The aim of this study was therefore to investigate the magnitude of signal errors in³¹P MRS when using ISIS. The results from the measurements with a homogeneous head phantom are as follows: at low TR/T1 ratios the contamination increases rapidly, especially for small (< 27 cm³) VOI sizes; at TR/T1 = 1, the signal from a 27 cm³ VOI was 20% too high, and from an 8 cm³ VOI 150% too high. The signal obtained from different VOI positions varied between 80 and 127%. The signal varied linearly with the³¹P concentration in the object. However, a too high signal was obtained when the concentration was lower in the region of interest (inner container) than in the rest of the phantom. The agreement between the simulations and measurements shows that the results of this study are generally applicable to the measurement geometry and the ISIS experiment order rather than being specific for the MR system studied. The errors obtained both experimentally and in computer simulations are too large to be ignored in clinical studies using the ISIS pulse sequence.     

Glycolytic atp production estimated from31p magnetic resonance spectroscopy measurements during ischemic exercisein vivo

In an oxygen-depleted muscle, glycolytically produced ATP is inversely related to the ([ATP] + creatine phosphate [PCr]) decrease because ATP, PCr, and glycolysis are virtually the only energy sources under these conditions. In particular, the onset of glycolysis or any appreciable increase in the rate of glycolytic ATP production will lead to a slower rate of ([ATP] + [PCr]) breakdown at a given energy consumption. To quantify this relationship, endurance athletes performed isometric foot plantar flexion (20% of a test force [TF],n=10; 50% TF,n=5) during local arterial occlusion. Parameters of energy metabolism were measured with³¹P magnetic resonance spectroscopy (³¹P-MRS). During exercise, [PCr] decreased to 80±10 (20% TF) and 11±4% (50% TF) of its resting concentration, and pH dropped from 7.04 0.01 to 6.98±0.10 (20% TF) and from 7.03±0.02 to 6.70 0.10 (50% TF). In both experiments, two phases of ([ATP] + [PCr]) decrease were observed: an initial faster decrease was followed by a slower decline. The latter phase started at about the time when the pH began to drop. The difference between a line extrapolated from the slope of the initial phase and the measured ([ATP] + [PCr]) decrease was used as an estimate for glycolytically produced ATP. This estimate and pH were significantly correlated withr=–0.97 (20% TF) andr=–0.99 (50% TF). These results indicate that glycolytically produced ATP can be estimated from the ([ATP] + [PCr]) decrease during exercise.     

Signal enhancement through heteronuclear polarisation transfer in in-vivo <Superscript>31</Superscript>P MR spectroscopy of the human brain

Significant ³¹P NMR signal enhancement through heteronuclear polarisation transfer was obtained in model solutions and in vivo on a 1.5-T whole-body MR scanner equipped with two RF channels. The much higher population differences involved in proton Zeeman energy levels can be transferred to the ³¹P levels with the refocused INEPT (insensitive nucleus enhancement by polarisation transfer) double-resonance experiment by means of a series of simultaneously applied broadband RF pulses. INEPT achieves a polarisation transfer from ¹H to ³¹P spin states by directly reordering the populations in spin systems with heteronuclear scalar coupling. Thus, only the ³¹P NMR signal of metabolites with scalar ¹H–³¹P coupling is amplified, while the other metabolite signals in the spectra are suppressed. Compared to Ernst-angle excitation, a repetition-time-dependent signal enhancement of η=(29±3)% for methylene diphosphonic acid (MDPA) and η=(56±1)% for phosphorylethanolamine (PE) was obtained on model solutions through optimisation of the temporal parameters of the pulse experiment. The results are in good agreement with numerical calculations of the theoretical model for the studied spin systems. With optimised echo times, in-vivo ³¹P signal enhancement of the same order was obtained in studies of the human brain.     

31P MR spectroscopy and in vitro markers of oxidative capacity in type 2 diabetes patients

Background: Skeletal muscle mitochondrial function in type 2 diabetes (T2D) is currently being studied intensively. In vivo ³¹P magnetic resonance spectroscopy (³¹P MRS) is a noninvasive tool used to measure mitochondrial respiratory function (MIFU) in skeletal muscle tissue. However, microvascular co-morbidity in long-standing T2D can interfere with the ³¹P MRS methodology. Aim: To compare ³¹P MRS-derived parameters describing in vivo MIFU with an in vitro assessment of muscle respiratory capacity and muscle fiber-type composition in T2D patients. Methods: ³¹P MRS was applied in long-standing, insulin-treated T2D patients. ³¹P MRS markers of MIFU were measured in the M. vastus lateralis. Muscle biopsy samples were collected from the same muscle and analyzed for succinate dehydrogenase activity (SDH) and fiber-type distribution. Results: Several ³¹P MRS parameters of MIFU showed moderate to good correlations with the percentage of type I fibers and type I fiber-specific SDH activity (Pearson’s R between 0.70 and 0.75). In vivo and in vitro parameters of local mitochondrial respiration also correlated well with whole-body fitness levels (VO _2peak) in these patients (Pearson’s R between 0.62 and 0.90). Conclusion: Good correlations exist between in vivo and in vitro measurements of MIFU in long-standing insulin-treated T2D subjects, which are qualitatively and quantitatively consistent with previous results measured in healthy subjects. This justifies the use of ³¹P MRS to measure MIFU in relation to T2D.     

31P-NMR spectroscopy of human blood and serum: first results from volunteers and patients with congestive heart failure,diabetes mellitus and hyperlipidaemia

³¹P-containing metabolites in human blood, serum and erythrocytes were measured or calculated. Phosphodiesters were found in serum, but not in erythrocytes. 2,3-diphosphoglycerate and 2,3-diphosphoglycerate/ATP ratios were increased in patients with congestive heart failure (2,3-diphosphoglycerate by 13% in mild to moderate, 31% in severe congestive heart failure, 2,3-diphosphoglycerate/ATP ratio by 9% in mild to moderate, 38% in severe congestive heart failure); phosphodiesters were increased in diabetes mellitus (by 26%) and even more so in hyperlipidaemia (by 57%). Changes of blood³¹P compounds with disease states may have diagnostic potential and should be recognized for correction of organ spectra.     

Physiological constraints on changes in pH and phosphorus metabolite concentrations in ischemically exercising muscle: implications for metabolic control and for the interpretation of31P-magnetic resonance spectroscopic studies

Relationships between pH and the concentrations of phosphocreatine (PCr), inorganic phosphate (Pi), and lactate during ischemic exercise depend on passive buffering, proton consumption as a consequence of net PCr breakdown, the control of glycogenolysis, (particularly in relation to the concentration of Pi, a substrate of glycogen phosphorylase that is produced by net PCr breakdown), and the creatine kinase equilibrium. The author analyzes the implications of these relationships for the interpretation of³¹P-magnetic resonance spectroscopic data and for the control of glycogenolysis. For realistic adenosine diphosphate (ADP) concentrations, given the constraints of the creatine kinase equilibrium, the pH must be near-linear with lactate, with an apparent buffer capacity (i.e., the ratio of lactate accumulation to pH change) that is nearly twice the true buffer capacity (i.e., the ratio of net proton loading to pH change). The implications for glycogenolytic control depend on adenosine triphosphate (ATP) turnover, but an upper limit of activation of glycogen phosphorylase (i.e., the amount of thea form) that would permit no increase in ADP concentration can be calculated. Phosphorylase activation during ischemic exercise seems approximately proportional to the power output, consistent with calcium stimulation of phosphorylaseb kinase. In simulations, ADP concentration is highly sensitive to this proportionality, as (unlike in purely oxidative exercise) ADP concentration is not known to participate in any closed feedback loops in ischemic exercise.     

In vivo1H-MR spectroscopy of the human heart

4. Conclusions Combined respiratory and cardiac triggering improves the localization accuracy and spectral quality in cardiac¹H-MRS dramatically leading to substantially increased spectral reproducibility. The best practical realization of double triggering turned out to be the use of the ECG amplitude when making use of the fact that it is modulated by respiration. In spite of the spectral quality achieved in most subjects, we still fail to record satisfactory spectra in a minority of subjects. The reasons for this are not understood at present but must be some particulars of either a given subject or the experimental setup. The cardiac¹H-MR spectra contain quantifiable contributions from creatine, TMA, lipids, and probably taurine. It is possible that the spectral contributions of creatine are subject to dipolar coupling similar to the observations for skeletal muscle.     

Creatine loading and resting skeletal muscle phosphocreatine flux: a saturation-transfer NMR study

³¹P saturation-transfer nuclear magnetic resonance spectroscopy was used to study skeletal muscle phosphocreatine (PCr) flux in healthy male volunteers. Data analysis included consideration of effects from incomplete saturation and radiofrequency spillover. Spectra were recorded from the resting gastrocnemius muscle before and after 6 days of creatine monohydrate (Cr·H₂O) intake (20 g/day). Parallel to an improved muscle performance during maximal intermittent exercise following Cr·H₂O supplementation, the concentration of PCr increased(P = 0.01) by 23% (34.9 ± 2.8 mmol/ 1 vs. 28.6 + 2.7 mmol/1), whereas other metabolites were unaffected (inorganic phosphate: 4.3± 1.4 mmol/1, free intracellular Mg²⁺: 1.1 ±0.7 mmol 1, cytosolic pH: 7.04 ± 0.02). Forward and reverse fluxes through the creatine kinase (CK) reaction did not change significantly from their baseline levels (v_for: 11.8 ± 5.4 mmol/1 per second vs. 15.3 ± 6.8 mmol 1 per second. (v_rev: 9.5 ± 3.4 mmol/1 per second vs. 10.9 ± 3.7 mmol/ 1 per second). The rate of PCr resynthesis in resting muscle is not limited by the CK reaction, which is near equilibrium. Consequently, the post-load increase in total creatine has no effect on the unidirectional CK reaction rates.     

Experimental31p nmr study of the influence of ionic strength on the apparent dissociation constant of mgatp

The classical method for³¹P NMR determination of intracellular free magnesium concentration ([Mg _free ²⁺ ]) requires an accurate knowledge of the apparent dissociation constant (K _D ) of MgATP. There is a large difference between the previously determined values ofK _D . Although the value of 50 µM, determined by a³¹P NMR method, is now largely accepted, a value of 86 µM has more recently been measured with a fitting method derived from the original one, and with a different ionic strength. The purpose of our study was to assess if the cause of the difference between these two previously reportedK _D values was due to the measuring method or to the ionic strength value used.Working at pH=7.2,T=37°C, and [KCl]=0.25 M, we performedK _D measurements with the original³¹P NMR method and with the fitting method. The results (67±13 µM and 61±20 µM, respectively) were not significantly different. Then, with the first method, we measured K_D at [KCl]=0.12 M and found a value of 19±5 µM. We conclude that the main cause of difference between theK _D values measured by³¹P NMR reside in the disparity of ionic strength values used for their measurement. OurK _D measurements at [KCl]=0.25 and 0.12 M demonstrate the importance of the ionic strength value used for imitating the intracellular medium on the absolute value of ([Mg _free ²⁺ ]) measured by³¹P NMR spectroscopy.Address for correspondence: Université Catholique de Louvain, Unité CPMC, Bâtiment Lavoisier, Place Louis Pasteur n°1, B-1348 Louvain-la-Neuve, Belgium. Additional reprints of this chapter may be obtained from the Reprints Department, Chapman & Hall, One Penn Plaza, New York, NY 10119.     

Contribution of various substrates to total citric acid cycle flux and ]anaplerosis as determined by13C isotopomer analysis and O2 consumption in the heart

A simple relationship between parameters derived from a¹³C NMR isotopomer analysis and O₂ consumption is presented that allows measurement of the absolute rate of acetyl-CoA oxidation and anaplerotic flux in tissues oxidizing a mixture of four substrates. The method was first applied in a study of the effects of work state and -adrenergic stimulation on net acetate oxidation and anaplerosis in the isolated working rat heart. The results demonstrate that the anticipated ratio of 2 between O₂ consumption and TCA cycle flux for hearts oxidizing only acetate holds at low workload when anaplerosis is low, but deviates toward a factor of 3 under high workload conditions when anaplerosis is increased. This analysis was also extended to hearts that oxidize a more physiological mixture of substrates including long-chain fatty acids, acetoacetate, lactate, pyruvate, and glucose. We show that the contribution each substrate makes to total TCA cycle flux can be determined by combined¹³C NMR and O₂ consumption measurements. The present study also demonstrates that stimulation of anaplerosis (by addition of propionate) can significantly alter the relative contribution each substrate makes to total TCA cycle flux. We conclude that if¹³C labeling patterns are selected appropriately, a comprehensive picture of flux through all major metabolic pathways feeding the cycle can be determined in a single experiment even when complex physiological mixtures of substrates are provided.     

Changes in proton transverse relaxation times of rat myocardium that has suffered a previous oxidative insult

An oxidative insult can induce severe damage, as in the phenomenon of myocardial ischemia and reperfusion. However, there are situations in which the damage is not so obvious (e.g., silent ischemia or aging), and the negative effects will be seen only in time. Our aim was to reveal these small changes in the myofilaments by using the nuclear magnetic resonance (NMR) technique. We used Wistar rat hearts in a constant-pressure Langendorff system, perfused with oxygenated Krebs-Henseleit buffer at 37°C. After 15 minutes of stabilization, the hearts were perfused with buffer supplemented with H₂O₂ at 50, 75, or 100 μmol/L for 15 or 30 minutes. Fifteen-minute and 45-minute perfusion controls and unperfused hearts were also collected. Heart rate (HR) and left ventricular developed pressure (LVDP) were determined with the help of a latex balloon, inserted in the left ventricle and connected with a pressure transducer. Proton transverse relaxation times (T ₂) were determined at the end of the experiment.T ₂ values were measured again in the same tissue fragments after they had been glycerinated and incubated in relaxation and rigor media. The functional parameters (HR, LVDP, coronary flow) were not significantly changed in control and 50 μmol/L H₂O₂ groups but were increased in the 75 μmol/L H₂O₂ group and significantly decreased in the 100 μmol/L H₂O₂ group.T ₂ is significantly decreased in rigor media starting with 50 μmol/L H₂O₂ administrated for 30 minutes and does not correlate with dose and duration of the oxidative insult.T ₂ in rigor is shorter than in relaxation media within the groups, and this difference is increased in the treated hearts.     

In vivo localized1 H spectroscopy of the rat eye at 7 T: preliminary studies

Preliminaryin vivo proton spectroscopic studies of the posterior chamber of the rat eye have been undertaken at 7 T. The Spatial and Chemical shift encoded Excitation (SPACE) localization sequence was used to acquire signals from 10-µl voxels and demonstrate the presence of metabolites associated with the vitreous humor, lens, retina, and the optic nerve. LocalizedT ₂ andT ₁ measurements of water in the vitreous humor indicate a relatively fluid environment. Susceptibility maps are used to demonstrate the difficulties ofin vivo spectroscopic investigations in the anterior regions of the eye. Comments are made concerning the implications for spectral resolution in these regions.     

Hyperammonemia and chronic hepatic encephalopathy: an in vivo PMRS study of the rat brain

The brain energy metabolism of rats affected by chronic hepatic encephalopathy due to portacaval shunting was monitored by in vivo³¹P-nuclear magnetic resonance spectroscopy before and after ammonium acetate administration. With respect to healthy unoperated and to sham operated controls, portacaval shunting decreased the levels of the nuclear magnetic resonance (NMR) visible brain phosphocreatine and nucleoside phosphates, and the intracellular [free Mg²⁺]. Ammonium acetate induced a further decrease of the levels of the NMR detectable phosphocreatine and nucleoside triphosphates and of the [free Mg²⁺], while the PMR spectra of the brain of non-shunted rats did not show any significant change even after treatment with ammonium acetate.     

Magnetic resonance imaging and67ga scan versus computed tomography in the staging and in the monitoring of mediastinal malignant lymphoma: a prospective pilot study

Purpose: To assess the potential value of magnetic resonance imaging (MRI) combined with⁶⁷Ga single-photon emission computed tomography (SPECT) versus computed tomography (CT) in the staging and in the monitoring of mediastinal malignant lymphoma. Materials and methods: Twenty-three patients, referred to our institute for the evaluation of lymphoma, underwent CT,⁶⁷Ga scan, and MRI between April 1993 and February 1996 at sequential intervals. The tests studied (MRI,^67Ga, and CT) were performed according to the following schedule: 1) before start of therapy; 2) after four courses of chemotherapy; and 3) 2, 6, 12, and 18 months after the end of treatment. Results: All patients studied at the time of diagnosis had abnormal gallium accumulation in the mediastinum as well as pathologic CT and pathologic signal intensity at MRI. Six months after the end of treatment full consistency was found between the results of MRI and SPECT, whereas during treatment and 2 months after the end of therapy MRI and⁶⁷Ga scan were not in agreement in nine patients. In the 23 patients in follow-up, in CT there were nine false-positive and three false-negative findings; in SPECT three false negatives; in MRI one false positive and one false negative. Conclusion: MRI can give morphologic information similar to CT, even superior due to multiplanarity and with major precision in the distinction between fibrosis and active disease. MRI is thus an alternative to CT. The association with SPECT allows a great diagnostic accuracy in the positive and negative predictive value.     

Measurement of arterial input function of <Superscript>17</Superscript>O water tracer in rat carotid artery by using a region-defined (REDE) implanted vascular RF coil

A method of determining arterial input function (AIF) by continuously detecting the ¹⁷O MR signal changes of ¹⁷O-labeled water tracer in the rat carotid artery using a region-defined (REDE) implanted vascular RF coil at 9.4 Tesla is reported. This coil has a compact physical size of 1 mm inner diameter, 3 mm outer diameter and 11 mm in length. It can be readily implanted into the rat neck and wrapped around the rat carotid artery for achieving adequate MR detection sensitivity for determining AIF with minimal surgical trauma. Water phantom and in vivo MR experiments were conducted for validating the coil's performance. A signal-to-noise ratio of ~20:1 was achieved for the ¹⁷O signal acquired from naturally abundant H₂ ¹⁷O in a small amount of blood (~7 μl) inside the rat carotid artery with an acquisition time of 11 s. The REDE RF coil design electromagnetically isolates the rat carotid artery from surrounding tissues and ensures that the MR signal detected by the RF coil is only attributable to the artery blood. It also minimizes the electromagnetic coupling between the implanted RF coil and a head surface coil tuned at the same operating frequency (two-coil configuration). This configuration allowed simultaneous measurements of dynamic changes of ¹⁷O MR signal of the H₂ ¹⁷O tracer in both rat carotid artery and brain. Compared to most contemporary MR approaches, the REDE implanted RF provides a simple, accurate, and promising solution for determination of AIF in small experimental animals.