Evidence of 13 C-lactate oxidation in the human brain from hyperpolarized 13 C-MRI.

PURPOSE: To test the hypothesis that lactate oxidation contributes to the 13 $$ {}^{13} $$ C-bicarbonate signal observed in the awake human brain using hyperpolarized 13 $$ {}^{13} $$ C MRI. METHODS: Healthy human volunteers (N = 6) were scanned twice using hyperpolarized 13 $$ {}^{13} $$ C-MRI, with increased radiofrequency saturation of 13 $$ {}^{13} $$ C-lactate on one set of scans. 13 $$ {}^{13} $$ C-lactate, 13 $$ {}^{13} $$ C-bicarbonate, and 13 $$ {}^{13} $$ C-pyruvate signals for 132 brain regions across each set of scans were compared using a clustered Wilcoxon signed-rank test. RESULTS: Increased 13 $$ {}^{13} $$ C-lactate radiofrequency saturation resulted in a significantly lower 13 $$ {}^{13} $$ C-bicarbonate signal (p = 0.04). These changes were observed across the majority of brain regions. CONCLUSION: Radiofrequency saturation of 13 $$ {}^{13} $$ C-lactate leads to a decrease in 13 $$ {}^{13} $$ C-bicarbonate signal, demonstrating that the 13 $$ {}^{13} $$ C-lactate generated from the injected 13 $$ {}^{13} $$ C-pyruvate is being converted back to 13 $$ {}^{13} $$ C-pyruvate and oxidized throughout the human brain.

First in-human evaluation of [1-13C]pyruvate in D2O for hyperpolarized MRI of the brain: A safety and feasibility study.

PURPOSE: To investigate the safety and value of hyperpolarized (HP) MRI of [1-13C]pyruvate in healthy volunteers using deuterium oxide (D2O) as a solvent. METHODS: Healthy volunteers (n = 5), were injected with HP [1-13C]pyruvate dissolved in D2O and imaged with a metabolite-specific 3D dual-echo dynamic EPI sequence at 3T at one site (Site 1). Volunteers were monitored following the procedure to assess safety. Image characteristics, including SNR, were compared to data acquired in a separate cohort using water as a solvent (n = 5) at another site (Site 2). The apparent spin-lattice relaxation time (T1) of [1-13C]pyruvate was determined both in vitro and in vivo from a mono-exponential fit to the image intensity at each time point of our dynamic data. RESULTS: All volunteers completed the study safely and reported no adverse effects. The use of D2O increased the T1 of [1-13C]pyruvate from 66.5 ± 1.6 s to 92.1 ± 5.1 s in vitro, which resulted in an increase in signal by a factor of 1.46 ± 0.03 at the time of injection (90 s after dissolution). The use of D2O also increased the apparent relaxation time of [1-13C]pyruvate by a factor of 1.4 ± 0.2 in vivo. After adjusting for inter-site SNR differences, the use of D2O was shown to increase image SNR by a factor of 2.6 ± 0.2 in humans. CONCLUSIONS: HP [1-13C]pyruvate in D2O is safe for human imaging and provides an increase in T1 and SNR that may improve image quality.

Current methods for hyperpolarized [1-13C]pyruvate MRI human studies.

MRI with hyperpolarized (HP) 13C agents, also known as HP 13C MRI, can measure processes such as localized metabolism that is altered in numerous cancers, liver, heart, kidney diseases, and more. It has been translated into human studies during the past 10 years, with recent rapid growth in studies largely based on increasing availability of HP agent preparation methods suitable for use in humans. This paper aims to capture the current successful practices for HP MRI human studies with [1-13C]pyruvate-by far the most commonly used agent, which sits at a key metabolic junction in glycolysis. The paper is divided into four major topic areas: (1) HP 13C-pyruvate preparation; (2) MRI system setup and calibrations; (3) data acquisition and image reconstruction; and (4) data analysis and quantification. In each area, we identified the key components for a successful study, summarized both published studies and current practices, and discuss evidence gaps, strengths, and limitations. This paper is the output of the "HP 13C MRI Consensus Group" as well as the ISMRM Hyperpolarized Media MR and Hyperpolarized Methods and Equipment study groups. It further aims to provide a comprehensive reference for future consensus, building as the field continues to advance human studies with this metabolic imaging modality.

Age-associated change in pyruvate metabolism investigated with hyperpolarized 13 C-MRI of the human brain.

In this study, hyperpolarized 13 C MRI (HP-13 C MRI) was used to investigate changes in the uptake and metabolism of pyruvate with age. Hyperpolarized 13 C-pyruvate was administered to healthy aging individuals (N = 35, ages 21-77) and whole-brain spatial distributions of 13 C-lactate and 13 C-bicarbonate production were measured. Linear mixed-effects regressions were performed to compute the regional percentage change per decade, showing a significant reduction in both normalized 13 C-lactate and normalized 13 C-bicarbonate production with age: - 7 % ± 2 % per decade for 13 C-lactate and - 9 % ± 4 % per decade for 13 C-bicarbonate. Certain regions, such as the right medial precentral gyrus, showed greater rates of change while the left caudate nucleus had a flat 13 C-lactate versus age and a slightly increasing 13 C-bicarbonate versus age. The results show that both the production of lactate (visible as 13 C-lactate signal) as well as the consumption of monocarboxylates to make acetyl-CoA (visible as 13 C-bicarbonate signal) decrease with age and that the rate of change varies by brain region.

Cardiac metabolic imaging using hyperpolarized [1-13 C]lactate as a substrate.

Hyperpolarized (HP) [1-13 C]lactate is an attractive alternative to [1-13 C]pyruvate as a substrate to investigate cardiac metabolism in vivo: it can be administered safely at a higher dose and can be polarized to a degree similar to pyruvate via dynamic nuclear polarization. While 13 C cardiac experiments using HP lactate have been performed in small animal models, they have not been demonstrated in large animal models or humans. Utilizing the same hardware and data acquisition methods as the first human HP 13 C cardiac study, 13 C metabolic images were acquired following injections of HP [1-13 C]lactate in porcine hearts. Data were also acquired using HP [1-13 C]pyruvate for comparison. The 13 C bicarbonate signal was localized to the myocardium and had a similar appearance with both substrates for all animals. No 13 C pyruvate signal was detected in the experiments following injection of HP 13 C lactate. The signal-to-noise ratio (SNR) of injected lactate was 88 ± 4% of the SNR of injected pyruvate, and the SNR of bicarbonate in the experiments using lactate as the substrate was 52 ± 19% of the SNR in the experiments using pyruvate as the substrate. The lower SNR was likely due to the shorter T1 of [1-13 C]lactate as compared with [1-13 C]pyruvate and the additional enzyme-catalyzed metabolic conversion step before the 13 C nuclei from [1-13 C]lactate were detected as 13 C bicarbonate. While challenges remain, the potential of HP lactate as a substrate for clinical metabolic imaging of human heart has been demonstrated.

Predicting response to radiotherapy of intracranial metastases with hyperpolarized [Formula: see text]C MRI.

BACKGROUND: Stereotactic radiosurgery (SRS) is used to manage intracranial metastases in a significant fraction of patients. Local progression after SRS can often only be detected with increased volume of enhancement on serial MRI scans which may lag true progression by weeks or months. METHODS: Patients with intracranial metastases (N = 11) were scanned using hyperpolarized [Formula: see text]C MRI prior to treatment with stereotactic radiosurgery (SRS). The status of each lesion was then recorded at six months post-treatment follow-up (or at the time of death). RESULTS: The positive predictive value of [Formula: see text]C-lactate signal, measured pre-treatment, for prediction of progression of intracranial metastases at six months post-treatment with SRS was 0.8 [Formula: see text], and the AUC from an ROC analysis was 0.77 [Formula: see text]. The distribution of [Formula: see text]C-lactate z-scores was different for intracranial metastases from different primary cancer types (F = 2.46, [Formula: see text]). CONCLUSIONS: Hyperpolarized [Formula: see text]C imaging has potential as a method for improving outcomes for patients with intracranial metastases, by identifying patients at high risk of treatment failure with SRS and considering other therapeutic options such as surgery.

Modeling, optimization, and validation of an extended-depth-of-field optical coherence tomography probe based on a mirror tunnel.

The diagnostic capability of high-resolution optical coherence tomography (OCT) may be enhanced by using extended depth-of-field (EDOF) imaging that retains high transverse resolution over long depths. A recently developed mirror-tunnel optical probe design (single-mode fiber to multimode fiber to lens structure) that generates coaxially focused modes has been previously shown to enable EDOF for endoscopic OCT applications. Here, we present ray-tracing optical modeling of this optical configuration, which has the potential to guide performance improvement through optimization. The Huygens wave propagation of the field was traced through probe components with initial lengths. The irradiance along the x-z plane was analyzed, yielding an average full width at half-maximum (FWHM) of 9 µm over a 640 µm DOF (defined as the axial range over which the beam's transverse FWHM is maintained). A custom merit function was then defined, based on the focal region illumination intensity profile that yielded the maximum possible depth having a constrained FWHM size. An orthogonal gradient descent optimization algorithm was then applied using this merit function, using the multimode fiber, spacer, and lens lengths as variables. Optimization resulted in a modeled mean 6 µm FWHM spot diameter over an EDOF of 1 mm. Following optimization, a probe was fabricated, and was validated using a custom-built near-field scanning pinhole beam profiler. The experimental results (6 µm mean FWHM over 800 µm EDOF) showed reasonable correspondence to the simulated predictions, demonstrating the potential utility of optical modeling and optimization for improving EDOF performance in mirror-tunnel endoscopic OCT probes.

Distinguishing selective mutism and social anxiety in children: a multi-method study.

Selective mutism (SM) is an anxiety disorder in which a child fails to speak in some situations (e.g., school) despite the ability to speak in other situations (e.g., home). Some work has conceptualized SM as a variant of social anxiety disorder (SAD) characterized by higher levels of social anxiety. Here, we empirically tested this hypothesis to see whether there were differences in social anxiety (SA) between SM and SAD across behavioral, psychophysiological, self-, parent-, and teacher-report measures. Participants included 158 children (Mage = 8.76 years, SD = 3.23) who were classified into three groups: children with SM and who were also highly socially anxious (SM + HSA; n = 48), highly socially anxious children without SM (HSA; n = 48), and control children (n = 62). Children participated in a videotaped self-presentation task, following which observed SA behaviors were coded, and salivary cortisol reactivity was measured. We also collected child, parent, and teacher reports of children's trait SA symptoms. The SM + HSA and HSA groups had similar observed non-verbal SA behavior, cortisol reactivity, and trait SA symptom levels according to parent and child reports, but SM + HSA children had significantly higher SA according to teacher report and observer-rated verbal SA behavior relative to the HSA group. As expected, control children had lower cortisol reactivity and SA across all measures relative to the other groups. Although SM and SAD in children share many similarities, SM may be characterized by greater SA in certain social contexts (e.g., school) and is distinguishable from SAD on behavioral measures of verbal SA.

Lactate topography of the human brain using hyperpolarized 13C-MRI.

Lactate is now recognized as an important intermediate in brain metabolism, but its role is still under investigation. In this work we mapped the distribution of lactate and bicarbonate produced from intravenously injected 13C-pyruvate over the whole brain using a new imaging method, hyperpolarized 13C MRI (N = 14, ages 23 to 77). Segmenting the 13C-lactate images into brain atlas regions revealed a pattern of lactate that was preserved across individuals. Higher lactate signal was observed in cortical grey matter compared to white matter and was highest in the precuneus, cuneus and lingual gyrus. Bicarbonate signal, indicating flux of [1-13C]pyruvate into the TCA cycle, also displayed consistent spatial distribution. One-way ANOVA to test for significant differences in lactate among atlas regions gave F = 87.6 and p < 10-6. This report of a "lactate topography" in the human brain and its consistent pattern is evidence of region-specific lactate biology that is preserved across individuals.

Partial Fourier reconstruction for improved resolution in 3D hyperpolarized 13 C EPI.

PURPOSE: Asymmetric in-plane k-space sampling of EPI can reduce the minimum achievable TE in hyperpolarized 13C with spectral-spatial radio frequency pulses, thereby reducing T2* weighting and signal-losses. Partial Fourier image reconstruction exploits the approximate Hermitian symmetry of k-space data and can be applied to asymmetric data sets to synthesize unmeasured data. Here we tested whether the application of partial Fourier image reconstruction would improve spatial resolution from hyperpolarized [1- 13C ]pyruvate scans in the human brain. METHODS: Fifteen healthy control subjects were imaged using a volumetric dual-echo echo-planar imaging sequence with spectral-spatial radio frequency excitation. Images were reconstructed by zero-filling as well as with the partial Fourier reconstruction algorithm projection-on-convex-sets. Resulting images were quantitatively evaluated with a no-reference image quality assessment. RESULTS: The no-reference image sharpness metric agreed with perceived improvements in image resolution and contrast. The [1- 13C ]lactate images benefitted most, followed by the [1- 13C ]pyruvate images. The 13C -bicarbonate images were improved by the smallest degree, likely owing to relatively lower SNR. CONCLUSIONS: Partial Fourier imaging and reconstruction were shown to improve the sharpness and contrast of human HP 13C brain data and is a viable method for enhancing resolution.

Correlation of hyperpolarized 13 C-MRI data with tissue extract measurements.

Hyperpolarized (HP) 13C MRI provides the means to monitor lactate metabolism noninvasively in tumours. Since 13C -lactate signal levels obtained from HP 13C imaging depend on multiple factors, such as the rate of 13C substrate delivery via the vasculature, the expression level of monocarboxylate transporters (MCTs) and lactate dehydrogenase (LDH), and the local lactate pool size, the interpretation of HP 13C metabolic images remains challenging. In this study, ex vivo tissue extract measurements (i.e., NMR isotopomer analysis, western blot analysis) derived from an MDA-MB-231 xenograft model in nude rats were used to test for correlations between the in vivo 13C data and the ex vivo measures. The lactate-to-pyruvate ratio from HP 13C MRI was strongly correlated with [1- 13C ]lactate concentration measured from the extracts using NMR (R = 0.69, p < 0.05), as well as negatively correlated with tumour wet weight (R = -  0.60, p < 0.05). In this tumour model, both MCT1 and MCT4 expressions were positively correlated with wet weight ( ρ = 0.78 and 0.93, respectively, p < 0.01). Lactate pool size and the lactate-to-pyruvate ratio were not significantly correlated.

Noninvasive Immunometabolic Cardiac Inflammation Imaging Using Hyperpolarized Magnetic Resonance.

RATIONALE: Current cardiovascular clinical imaging techniques offer only limited assessment of innate immune cell-driven inflammation, which is a potential therapeutic target in myocardial infarction (MI) and other diseases. Hyperpolarized magnetic resonance (MR) is an emerging imaging technology that generates contrast agents with 10- to 20 000-fold improvements in MR signal, enabling cardiac metabolite mapping. OBJECTIVE: To determine whether hyperpolarized MR using [1-13C]pyruvate can assess the local cardiac inflammatory response after MI. METHODS AND RESULTS: We performed hyperpolarized [1-13C]pyruvate MR studies in small and large animal models of MI and in macrophage-like cell lines and measured the resulting [1-13C]lactate signals. MI caused intense [1-13C]lactate signal in healing myocardial segments at both day 3 and 7 after rodent MI, which was normalized at both time points after monocyte/macrophage depletion. A near-identical [1-13C]lactate signature was also seen at day 7 after experimental MI in pigs. Hyperpolarized [1-13C]pyruvate MR spectroscopy in macrophage-like cell suspensions demonstrated that macrophage activation and polarization with lipopolysaccharide almost doubled hyperpolarized lactate label flux rates in vitro; blockade of glycolysis with 2-deoxyglucose in activated cells normalized lactate label flux rates and markedly inhibited the production of key proinflammatory cytokines. Systemic administration of 2-deoxyglucose after rodent MI normalized the hyperpolarized [1-13C]lactate signal in healing myocardial segments at day 3 and also caused dose-dependent improvement in IL (interleukin)-1ß expression in infarct tissue without impairing the production of key reparative cytokines. Cine MRI demonstrated improvements in systolic function in 2-DG (2-deoxyglucose)-treated rats at 3 months. CONCLUSIONS: Hyperpolarized MR using [1-13C]pyruvate provides a novel method for the assessment of innate immune cell-driven inflammation in the heart after MI, with broad potential applicability across other cardiovascular disease states and suitability for early clinical translation.

Longitudinal trajectories of depression symptoms in children with epilepsy.

AIM: To examine self- and proxy-reported symptoms of depression in children with epilepsy. METHOD: This was a prospective longitudinal cohort study of children with epilepsy. Participants were treated at six Canadian tertiary-care centers and followed over 28 months with repeated assessments of child self-reported symptoms of depression using the Children's Depression Inventory Short-Form (CDI-S). Trajectories of symptoms of depression were estimated using linear mixed effects (LME) modeling. RESULTS: At baseline, 477 children had complete data (mean age [SD] 11y 5mo [2y 1mo], range 7y 7mo-15y 1mo; 234 females, 243 males). Mean CDI-S T score at baseline was 45.7 (SD=7.5) and at 28 months was 44.9 (SD=8.2), both were within the 'average' range. Results from LME modeling revealed mean raw CDI-S score of 1.897, corrected for age 10 years (corresponding to T scores slightly below the normed mean of 50), with no significant change over three measurements (slope=-0.113, p=0.135), indicating that CDI-S scores were stable over 28 months. Children with high initial CDI-S scores had lower subsequent scores, as demonstrated by the correlation of -0.827 between intercept and slope (p<0.001). Parents reported comparable findings. INTERPRETATION: Self- and proxy-reported symptoms of depression were generally low and stable over an extended follow-up period. Normalization of scores was seen upon repeated assessment, even in children with higher scores of symptoms of depression at one point. These findings speak to the value and importance of repeated assessment over time. WHAT THIS PAPER ADDS: In children with epilepsy, self- and proxy-reported symptoms of depression were generally low and stable over 28 months. The trajectory of symptoms of depression was not associated with seizure severity, whether considering the frequency or type of seizures. Parents' reports of symptoms of depression were comparable to the children's self-evaluations.

What Antibullying Program Designs Motivate Student Intervention in Grades 5 to 8?

Educators detect and intervene in a small proportion of bullying incidents. Although students are present when many bullying episodes occur, they are often reluctant to intervene. This study explored attributes of antibullying (AB) programs influencing the decision to intervene. Grade 5, 6, 7, and 8 students (N = 2,033) completed a discrete choice experiment examining the influence of 11 AB program attributes on the decision to intervene. Multilevel analysis revealed 6 latent classes. The Intensive Programming class (28.7%) thought students would intervene in schools with daily AB activities, 8 playground supervisors, mandatory reporting, and suspensions for perpetrators. A Minimal Programming class (10.3%), in contrast, thought monthly AB activities, 4 playground supervisors, discretionary reporting, and consequences limited to talking with teachers would motivate intervention. Membership in this class was linked to Grade 8, higher dispositional reactance, more reactance behavior, and more involvement as perpetrators. The remaining 4 classes were influenced by different combinations of these attributes. Students were more likely to intervene when isolated peers were included, other students intervened, and teachers responded quickly. Latent class analysis points to trade-offs in program design. Intensive programs that encourage intervention by students with little involvement as perpetrators may discourage intervention by those with greater involvement as perpetrators, high dispositional reactance, or more reactant behavior.

Trajectories of Observed Shyness and Psychosocial Adjustment in Children.

Shyness can manifest as inhibition, fear, and avoidance in the context of social novelty and situations of perceived social evaluation. In the present study, 130 children (Mage = 7.6 years, SD = 1.8) participated in a videotaped self-presentation task across three separate visits spanning approximately 3 years in early and middle childhood. Children's observed shyness was best characterized by two trajectories, including a high-stable class (19%) and a low-stable class (81%). Girls were more likely than boys to follow a pattern of high-stable observed shyness. Further, children in the high-stable observed shyness class were rated by parents and teachers as more socially anxious relative to children in the low-stable class, and boys in the high-stable observed shyness class were rated by their teachers as displaying more depressive symptoms relative to girls. These findings suggest that a subset of children display stable behavioral shyness, and this is correlated with psychosocial functioning.

Parent Proxy Discrepancy Groups of Quality of Life in Childhood Epilepsy.

OBJECTIVES: To study the extent to which parents are able to serve as true proxies for their children with epilepsy using a more granular approach than has been found in any study to date. METHODS: Proxy resemblance to the child was based on discrepancy in z-centered child minus parent scores of matching quality-of-life (QOL) domains for 477 dyads. Latent class mixed models (LCMMs) were built, with child's age as the independent variable for epilepsy-specific and generic QOL. Data were obtained from the QUALITÉ Canadian cohort, which recruited children with epilepsy aged 8 to 14 years at baseline and their parents. RESULTS: Both epilepsy-specific and generic LCMMs produced latent classes representing proxies that were overly positive, overly negative, or in agreement relative to their children with posterior probabilities of 79% to 84%. The "agreement" classes had N = 411 and N = 349 in the epilepsy-specific and generic LCMMs, respectively. The epilepsy-specific LCMM had a small unique class of N = 5 with a posterior probability of 88% called "growing discrepancy." CONCLUSIONS: Most parents of children with epilepsy can serve as valid proxies for their children on QOL scales. Poorer parental adaptation is more related to overly negative proxies, whereas low peer support from the child's perspective is more related to overly positive proxies.

Simultaneous multislice acquisition without trajectory modification for hyperpolarized 13 C experiments.

PURPOSE: To investigate the feasibility of performing large FOV hyperpolarized 13 C metabolic imaging using simultaneous multislice excitation. METHODS: A spectral-spatial multislice excitation pulse was constructed by cosine modulation and incorporated into a 13 C spiral imaging sequence. Phantom and in vivo pig experiments were performed to test the feasibility of simultaneous multislice data acquisition and image reconstruction. In vivo cardiac-gated images of hyperpolarized pyruvate, bicarbonate, and lactate were obtained at 1 × 1 × 1 cm3 resolution over a 48 × 48 × 24 cm3 FOV with 2-fold acceleration in the slice direction. Sensitivity encoding was used for image reconstruction with both autocalibrated and numerically calculated coil sensitivities. RESULTS: Simultaneous multislice images obtained with 2-fold acceleration were comparable to reference unaccelerated images. Retained SNR figures greater than 80% were achieved over the part of the image containing the heart. CONCLUSION: This method is anticipated to enable large FOV imaging studies using hyperpolarized 13 C substrates, with an aim toward whole-body exams that have to date been out of reach.

Dual-Echo EPI sequence for integrated distortion correction in 3D time-resolved hyperpolarized 13 C MRI.

PURPOSE: To provide built-in off-resonance correction in time-resolved, volumetric hyperpolarized 13 C metabolic imaging by implementing a novel dual-echo 3D echo-planar imaging (EPI) sequence and reconstruction. METHODS: A spectral-spatial pulse for single-resonance excitation followed by a dual-echo 3D EPI readout was implemented to provide 64 × 8 × 6 cm3 coverage at 5 × 5 × 5 mm3 nominal resolution. Multiple sources of EPI distortions were encoded using a multi-echo 1 H EPI reference scan. Phase maps computed from the reference scans were combined with a bulk 13 C frequency offset encoded in the dual-echo [1-13 C]pyruvate images to correct geometric distortion and improve spatial registration. The proposed scheme was validated in a phantom study, and in vivo [1-13 C]pyruvate and [1-13 C]lactate rat images were acquired with intentional transmit frequency deviations to assess the dual-echo 3D EPI sequence. RESULTS: The phantom study demonstrated improved spatial registration in off-resonance corrected images. Close agreement was observed between metabolic kidney signal and the underlying anatomy in rat imaging experiments. Relative to a single-echo acquisition, the coherent addition of the two corrected echoes provided the expected increase in signal-to-noise ratio by approximately 2. CONCLUSION: A novel dual-echo 3D EPI acquisition sequence for integrated off-resonance correction in hyperpolarized 13 C imaging was developed and demonstrated. The proposed sequence offers clear advantages over flyback EPI for time-resolved metabolic mapping. Magn Reson Med 79:643-653, 2018. © 2017 International Society for Magnetic Resonance in Medicine.

Probing the cardiac malate-aspartate shuttle non-invasively using hyperpolarized [1,2-13 C2 ]pyruvate.

Previous studies have demonstrated that using hyperpolarized [2-13 C]pyruvate as a contrast agent can reveal 13 C signals from metabolites associated with the tricarboxylic acid (TCA) cycle. However, the metabolites detectable from TCA cycle-mediated oxidation of [2-13 C]pyruvate are the result of several metabolic steps. In the instance of the [5-13 C]glutamate signal, the amplitude can be modulated by changes to the rates of pyruvate dehydrogenase (PDH) flux, TCA cycle flux and metabolite pool size. Also key is the malate-aspartate shuttle, which facilitates the transport of cytosolic reducing equivalents into the mitochondria for oxidation via the malate-α-ketoglutarate transporter, a process coupled to the exchange of cytosolic malate for mitochondrial α-ketoglutarate. In this study, we investigated the mechanism driving the observed changes to hyperpolarized [2-13 C]pyruvate metabolism. Using hyperpolarized [1,2-13 C]pyruvate with magnetic resonance spectroscopy (MRS) in the porcine heart with different workloads, it was possible to probe 13 C-glutamate labeling relative to rates of cytosolic metabolism, PDH flux and TCA cycle turnover in a single experiment non-invasively. Via the [1-13 C]pyruvate label, we observed more than a five-fold increase in the cytosolic conversion of pyruvate to [1-13 C]lactate and [1-13 C]alanine with higher workload. 13 C-Bicarbonate production by PDH was increased by a factor of 2.2. Cardiac cine imaging measured a two-fold increase in cardiac output, which is known to couple to TCA cycle turnover. Via the [2-13 C]pyruvate label, we observed that 13 C-acetylcarnitine production increased 2.5-fold in proportion to the 13 C-bicarbonate signal, whereas the 13 C-glutamate metabolic flux remained constant on adrenergic activation. Thus, the 13 C-glutamate signal relative to the amount of 13 C-labeled acetyl-coenzyme A (acetyl-CoA) entering the TCA cycle was decreased by 40%. The data strongly suggest that NADH (reduced form of nicotinamide adenine dinucleotide) shuttling from the cytosol to the mitochondria via the malate-aspartate shuttle is limited on adrenergic activation. Changes in [5-13 C]glutamate production from [2-13 C]pyruvate may play an important future role in non-invasive myocardial assessment in patients with cardiovascular diseases, but careful interpretation of the results is required.