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BACKGROUNDThe communication through coherence model posits that brain rhythms are synchronized across different frequency bands and that effective connectivity strength between interacting regions depends on their phase relation. Evidence to support the model comes mostly from electrophysiological recordings in animals while evidence from human data is limited.METHODSHere, an fMRI-EEG-TMS (fET) instrument capable of acquiring simultaneous fMRI and EEG during noninvasive single pulse TMS applied to dorsolateral prefrontal cortex (DLPFC) was used to test whether prefrontal EEG alpha phase moderates TMS-evoked top-down influences on subgenual, rostral and dorsal anterior cingulate cortex (ACC). Results in healthy volunteers (n=11) were compared to those from patients with major depressive disorder (MDD) (n=17) collected as part of a ongoing clinical trial investigation.RESULTSIn both groups, TMS-evoked functional connectivity between DLPFC and subgenual ACC (sgACC) depended on the EEG alpha phase. TMS-evoked DLPFC to sgACC effective connectivity (EC) was moderated by EEG alpha phase in healthy volunteers, but not in the MDD patients. Top-down EC was inhibitory for TMS onsets during the upward slope of the alpha wave relative to TMS timed to the downward slope of the alpha wave. Prefrontal EEG alpha phase dependent effects on TMS-evoked fMRI BOLD activation of the rostral anterior cingulate cortex were detected in the MDD patient group, but not in the healthy volunteer group.DISCUSSIONResults demonstrate that TMS-evoked top-down influences vary as a function of the prefrontal alpha rhythm, and suggest clinical applications whereby TMS is synchronized to the brain’s internal rhythms in order to more efficiently engage deep therapeutic targets.

Background Neonatal abstinence syndrome (NAS) is a significant problem. Opioid withdrawal induces oxidative stress and disrupts glutamate and glutathione homeostasis. We hypothesized that N-acetylcysteine (NAC) administered during acute opioid withdrawal in neonatal rats would decrease withdrawal behaviors and normalize CNS glutathione and glutamate. Methods Osmotic minipumps with methadone (opioid dependent, OD) and saline (Sham) were implanted into Sprague Dawley dams 7 days prior to delivery. Pups were randomized to receive either naloxone plus saline or NAC (50–100 mg/kg), administered on postnatal day (PND) 7. We performed MR spectroscopy on PND6–7 before, 30 min, and 120 min after withdrawal. On PND7, we assessed withdrawal behaviors for 90 min after naloxone administration and summed scores during peak withdrawal period. Results Mean summed behavioral scores were significantly different between groups (χ2 (2) = 10.49, p = 0.005) but not different between NAC/NAL/OD and Sham (p = 0.14): SAL/NAL/OD = 17.2 ± 4.2 (n = 10); NAC/NAL/OD = 11.3 ± 5.6 (n = 9); Sham = 6.5 ± 0.6 (n = 4). SAL/NAL/OD pups had decreased glutathione at 120 min (p = 0.01), while NAC/NAL/OD pups maintained pre-withdrawal glutathione (p = 0.26). Conclusion In antenatal OD, NAC maintains CNS glutathione and mitigates acute opioid withdrawal in neonatal rats. This is the first study to demonstrate acute opioid withdrawal neurochemical changes in vivo in neonatal OD. NAC is a potential novel treatment for NAS.

Although the neurotransmitters/modulators glutamate and, more recently, glycine have been implicated in the development and maintenance of Alcohol Use Disorder (AUD) in preclinical research, human proton magnetic resonance spectroscopy ((1)H-MRS) studies have focused solely on the measurement of glutamate. The purpose of the present analysis was to examine the relative associations of brain glutamate and glycine levels with recent heavy drinking in 41 treatment naïve individuals with AUD using (1)H-MRS. The present study is the first that we are aware of to report in vivo brain glycine levels from an investigation of addiction. Dorsal Anterior Cingulate Cortex (dACC) glutamate and glycine concentration estimates were obtained using Two-Dimensional J-Resolved Point Resolved Spectroscopy at 3 Tesla, and past 2-week summary estimates of alcohol consumption were assessed via the Timeline Followback method. Glutamate (β= −0.44, t= −3.09, p= 0.004) and glycine (β= −0.68, t= −5.72, p< 0.001) were each significantly, inversely associated with number of heavy drinking days when considered alone. However, when both variables were simultaneously entered into a single regression model, the effect of glutamate was no longer significant (β= −0.11, t= −0.81, p= 0.42) whereas the effect of glycine remained significant (β= −0.62, t= −4.38, p< 0.001). The present study extends the literature by demonstrating a unique, inverse association of brain glycine levels with recent heavy drinking in treatment naïve individuals with AUD. If replicated and extended, these data could lead to enhanced knowledge of how glycinergic systems change with alcohol consumption and AUD progression leading to pharmacological interventional/preventative strategies that modulate brain glycine levels.

The genomes of mitochondria and chloroplasts contain ribosomal RNA (rRNA) genes, reflecting their evolutionary ancestry as free-living bacteria prior to endosymbiosis. In microbiome studies of animals, plants, or other eukaryotic hosts, these organellar rRNAs are often amplified. If identified, they can be discarded, merely reducing sequencing depth. However, incorrectly annotated mitochondrial reads may compromise statistical analysis by distorting relative abundances of free-living microbes. We quantified this by reanalyzing 7,459 samples from seven 16S rRNA sequencing studies, including the microbiomes of 927 unique animal genera. We find that under-annotation of cryptic mitochondrial reads affects multiple of these large-scale cross-species microbiome comparisons, and can be severe in some samples. It also varies between host species, potentially biasing cross-species microbiome comparisons. We propose a straightforward solution: by supplementing existing taxonomies with diverse mitochondrial rRNA sequences, we resolve up to 97% of unique unclassified sequences in some entire studies as mitochondrial (14% averaged across all studies), without increasing false positive annotations in mitochondria-free mock communities. Overall, improved annotation decreases the proportion of unknown sequences by ≥10-fold in 2,262 of 7,459 samples (30%), including representatives from 5 of 7 studies examined. While standard DADA2 analyses are severely affected, the default positive filter in Deblur run through QIIME2 discards many divergent mitochondrial sequences, preventing bias in analysis, but also making analysis of these sequences more difficult. We recommend leveraging mitochondrial sequence diversity to better identify, remove and analyze mitochondrial rRNA gene sequences in microbiome studies.

An 11–25% increase in total ventricular volume has been documented in astronauts following spaceflight on the ISS. Given the approximately 2-year time interval between pre- and post-flight MRI, it is unknown if ventricular enlargement simply reflects normal aging or is unique to spaceflight exposure. Therefore, we compared percent ventricular volume change per year (PVVC/yr) documented on pre- to post-flight MRI in a group of NASA ISS astronauts (n = 18, 16.7% women, mean age (SD) 48.43 (4.35) years) with two groups who underwent longitudinal MRI: (1.) healthy age- and sex-matched adults (n = 18, 16.7% women, mean age (SD) 51.26 (3.88) years), and (2.) healthy older adults (n = 79, 16.5% women, mean age (SD) 73.26 (5.34) years). The astronauts, who underwent a mean (SD) 173.4 (51.3) days in spaceflight, showed a greater increase in PVVC/yr than the control (6.86 vs 2.23%, respectively, p

Perception and cognition in the brain are naturally characterized as spatiotemporal processes. Decision-making, for example, depends on coordinated patterns of neural activity cascading across the brain, running in time from stimulus to response and in space from primary sensory regions to the frontal lobe. Measuring this cascade is key to developing an understanding of brain function. Here we report on a novel methodology that employs multi-modal imaging for inferring this cascade in humans at unprecedented spatiotemporal resolution. Specifically, we develop an encoding model to link simultaneously measured electroencephalography (EEG) and functional magnetic resonance imaging (fMRI) signals to infer high-resolution spatiotemporal brain dynamics during a perceptual decision. After demonstrating replication of results from the literature, we report previously unobserved sequential reactivation of a substantial fraction of the pre-response network whose magnitude correlates with a proxy for decision confidence. Our encoding model, which temporally tags BOLD activations using time localized EEG variability, identifies a coordinated and spatially distributed neural cascade that is associated with a perceptual decision. In general the methodology illuminates complex brain dynamics that would otherwise be unobservable using fMRI or EEG acquired separately.

Background and purposeCurrently, there is neither a standard protocol for vessel wall MR imaging of intracranial atherosclerotic disease (ICAD) nor a gold standard phantom to compare MR sequences. In this study, a plaque phantom is developed and characterized that provides a platform for establishing a uniform imaging approach for ICAD.Materials and methodsA patient specific injection mold was 3D printed to construct a geometrically accurate ICAD phantom. Polyvinyl alcohol hydrogel was infused into the core shell mold to form the stenotic artery. The ICAD phantom incorporated materials mimicking a stenotic vessel and plaque components, including fibrous cap and lipid core. Two phantoms were scanned using high resolution cone beam CT and compared with four different 3 T MRI systems across eight different sites over a period of 18 months. Inter-phantom variability was assessed by lumen dimensions and contrast to noise ratio (CNR).ResultsQuantitative evaluation of the minimum lumen radius in the stenosis showed that the radius was on average 0.80 mm (95% CI 0.77 to 0.82 mm) in model 1 and 0.77 mm (95% CI 0.74 to 0.81 mm) in model 2. The highest CNRs were observed for comparisons between lipid and vessel wall. To evaluate manufacturing reproducibility, the CNR variability between the two models had an average absolute difference of 4.31 (95% CI 3.82 to 5.78). Variation in CNR between the images from the same scanner separated by 7 months was 2.5–6.2, showing reproducible phantom durability.ConclusionsA plaque phantom composed of a stenotic vessel wall and plaque components was successfully constructed for multicenter high resolution MRI standardization.

BACKGROUND AND PURPOSE: Widespread brain structural changes are seen following extended spaceflight missions. The purpose of this study was to investigate whether these structural changes are associated with alterations in motor or cognitive function. MATERIALS AND METHODS: Brain MR imaging scans of National Aeronautics and Space Administration astronauts were retrospectively analyzed to quantify pre- to postflight changes in brain structure. Local structural changes were assessed using the Jacobian determinant. Structural changes were compared with clinical findings and cognitive and motor function. RESULTS: Long-duration spaceflights aboard the International Space Station, but not short-duration Space Shuttle flights, resulted in a significant increase in total ventricular volume (10.7% versus 0%, P < .001, n = 12 versus n = 7). Total ventricular volume change was significantly associated with mission duration (r = 0.72, P = .001, n = 19) but negatively associated with age (r = −0.48, P = .048, n = 19). Long-duration spaceflights resulted in significant crowding of brain parenchyma at the vertex. Pre- to postflight structural changes of the left caudate correlated significantly with poor postural control; and the right primary motor area/midcingulate correlated significantly with a complex motor task completion time. Change in volume of 3 white matter regions significantly correlated with altered reaction times on a cognitive performance task (bilateral optic radiations, splenium of the corpus callosum). In a post hoc finding, astronauts who developed spaceflight-associated neuro-ocular syndrome demonstrated smaller changes in total ventricular volume than those who did not (12.8% versus 6.5%, n = 8 versus n = 4). CONCLUSIONS: While cautious interpretation is appropriate given the small sample size and number of comparisons, these findings suggest that brain structural changes are associated with changes in cognitive and motor test scores and with the development of spaceflight-associated neuro-optic syndrome.

Purpose Approximately 2 to 4% of the US population have been estimated to seek treatment for temporomandibular symptoms, predominately women. The study purpose was to determine whether sex-specific differences in temporomandibular morphometry result from scaling with sex differences in skull size and shape or intrinsic sex-specific differences. Materials and Methods A total of 22 (11 male [aged 74.5 ± 9.1 years]; 11 female [aged 73.6 ± 12.8 years]) human cadaveric heads with no history of temporomandibular disc derangement underwent cone beam computed tomography and high-resolution magnetic resonance imaging scanning to determine 3-dimensional cephalometric parameters and temporomandibular morphometric outcomes. Regression models between morphometric outcomes and cephalometric parameters were developed, and intrinsic sex-specific differences in temporomandibular morphometry normalized by cephalometric parameters were determined. Subject-specific finite element (FE) models of the extreme male and extreme female conditions were developed to predict variations in articular disc stress–strain under the same joint loading. Results In some cases, sex differences in temporomandibular morphometric parameters could be explained by linear scaling with skull size and shape; however, scaling alone could not fully account for some differences between sexes, indicating intrinsic sex-specific differences. The intrinsic sex-specific differences in temporomandibular morphometry included an increased condylar medial length and mediolateral disc lengths in men and a longer anteroposterior disc length in women. Considering the extreme male and female temporomandibular morphometry observed in the present study, subject-specific FE models resulted in sex differences, with the extreme male joint having a broadly distributed stress field and peak stress of 5.28 MPa. The extreme female joint had a concentrated stress field and peak stress of 7.37 MPa. Conclusions Intrinsic sex-specific differences independent of scaling with donor skull size were identified in temporomandibular morphometry. Understanding intrinsic sex-specific morphometric differences is critical to determining the temporomandibular biomechanics given the effect of anatomy on joint contact mechanics and stress–strain distributions and requires further study as one potential factor for the increased predisposition of women to temporomandibular disc derangement.

Proton Magnetic Resonance Spectroscopy ((1)H-MRS) studies have demonstrated abnormal levels of a variety of neurometabolites in treatment-seeking individuals with moderate-severe Alcohol Use Disorder (AUD) following acute withdrawal. In contrast, few studies have investigated neurochemical changes across early abstinence in less-severe, treatment-naïve AUD. The present study, which represents the primary report of a research grant from ABMRF/The Alcohol Research Fund, measured dorsal anterior cingulate cortex (dACC) GABA, glutamate, and glutamine levels in treatment-naïve AUD (n=23) via three (1)H-MRS scans spaced across a planned week of abstinence from alcohol. In addition to AUD participants, twelve light drinkers completed two scans, separated by 48 hours, to ensure that results in AUD were not produced by between-scan differences other than abstinence from alcohol. (1)H-MRS spectra were acquired in dACC at each scan using 2D J-resolved Point Resolved Spectroscopy. Linear mixed modeling results demonstrated a significant increase in GABA, but not glutamate or glutamine (ps=0.237–0.626), levels between scans 1 and 2 (+8.88%, p = 0.041), with no difference between scans 2 and 3 (+1.00%, p = 0.836), in AUD but not LD (F=1.24, p=0.290) participants. Exploratory regression analyses tentatively revealed a number of significant prospective associations between changes in glutamine levels and heavy drinking, craving and withdrawal symptoms. Most notably, the present study demonstrated return from abnormally-low to normal GABA levels in treatment-naïve AUD within 3d of their last drink; the pattern of results was consistent with glutamate and glutamine disturbances being exclusive to relatively more-severe AUD.

Little attention has been paid to relating MRS outputs of vendor‐supplied platforms to those from research software. This comparison is crucial to advance MRS as a clinical prognostic tool for disease or injury, recovery, and outcome. The work presented here investigates the agreement between metabolic ratios reported from vendor‐provided and LCModel fitting algorithms using MRS data obtained on Siemens 3 T TIM Trio and 3 T Skyra MRI scanners in a total of 55 premature infants and term neonates with hypoxic ischemic encephalopathy (HIE). We compared peak area ratios in single voxels placed in basal ganglia (BG) and frontal white matter (WM) using standard PRESS (TE = 30 ms and 270 ms) and STEAM (TE = 20 ms) MRS sequences at multiple times after birth from 5 to 60 days. A total of 74 scans met quality standards for inclusion, reflecting a spectrum of neonatal disease and several months of early infant development. For the long TE PRESS sequence, N‐acetylaspartate (NAA) and Choline (Cho) ratios to Creatine (Cr) correlated strongly between LCModel and vendor‐supplied software in the BG. For shorter TEs, the ratios of NAA/Cr and Cho/Cr were more closely related using STEAM at TE = 20 ms in BG and WM, which was significantly better than using PRESS at TE = 30 ms in the BG of HIE infants. At short TEs, however, it is still unclear which MRS sequence, STEAM or PRESS, is superior and thus more work is required in this regard for translating research‐generated MRS ratios to clinical diagnosis and prognostication, and unlocking the potential of MRS for in vivo metabolomics. MRS at both long and short TEs is desirable for standard metabolites such as NAA, Cho and Cr, along with important lower concentration metabolites such as myo‐inositol and glutathione.

Depression is a serious mental illness that is frequently resistant to a first round of pharmacotherapy. Electroconvulsive therapy (ECT) is effective even for such treatment resistant depression but is associated with significant adverse effects. Repetitive transcranial magnetic stimulation (rTMS) over the left dorsolateral prefrontal cortex in comparison causes only mild discomfort but is less effective than ECT. We hypothesize that TMS treatment efficacy could be improved by locking TMS onset to a specific, potentially subject specific phase of the prefrontal alpha rhythm in the electroencephalogram (EEG). Here, we present an instrument that can track and predict phase of the alpha rhythm in the EEG to precisely target TMS while concurrently recording functional magnetic resonance imaging (fMRI) to study local and distributed hemodynamic brain responses to stimulation. Tests of the instrument with three healthy adults indicate that EEG phase-locked TMS can be administered accurately enough to start testing systematically whether specific stimulation protocols can lead to clinically significant improvements in depression. To our knowledge, this is the first system that can deliver TMS phase-locked to the alpha rhythm while concurrently recording fMRI. For patients, such EEG guided TMS treatment could lead to better clinical outcomes and lower incidence of adverse effects.

Background Proton magnetic resonance spectroscopy (1 H-MRS) studies have demonstrated abnormal levels of a variety of neurometabolites in inpatients/outpatients with alcohol use disorder (AUD) following acute alcohol withdrawal relative to healthy controls. In contrast, few studies have compared neurometabolite levels between less severe, treatment-naive AUD individuals and light drinkers (LD) or related them to recent alcohol consumption. The present study compared neurometabolite levels between treatment-naive AUD and LD individuals. Methods Twenty treatment-naive individuals with AUD and 20 demographically matched LD completed an 1 H-MRS scan, approximately 2.5 days following their last reported drink. 1 H-MRS data were acquired in dorsal anterior cingulate (dACC) using a 2-dimensional J-resolved point-resolved spectroscopy sequence. dACC neurometabolite levels, with a focus on glutamate, glutamine, and GABA, were compared between AUD and LD participants. The associations between metabolite levels and recent drinking were explored. Results AUD participants had significantly lower concentrations of GABA (Cohen's d = 0.79, p = 0.017) and glutamine (Cohen's d = 1.12, p = 0.005), but not glutamate (Cohen's d = 0.05, p = 0.893), relative to LD. As previously reported, AUD participants' glutamate and N-acetylaspartate concentrations were inversely associated with their number of heavy drinking days. In contrast, neither number of drinking (mean p = 0.56) nor heavy drinking (mean p = 0.47) days were associated with metabolite concentrations in LD. Conclusions The present study demonstrated significantly lower levels of prefrontal γ-aminobutyric acid and glutamine in treatment-naive individuals with AUD relative to LD. Whether these findings reflect the neurotoxic consequence and/or neuroadaptive response of alcohol consumption versus a predrinking trait, and therefore a more durable neurochemical disturbance, awaits elucidation from longitudinal studies.

Therapeutic hypothermia does not improve outcomes in neonatal hypoxia ischemia (HI) complicated by perinatal infection, due to well-described, pre-existing oxidative stress and neuroinflammation that shorten the therapeutic window. For effective neuroprotection post-injury, we must first define and then target CNS metabolomic changes immediately after endotoxin-sensitized HI (LPS-HI). We hypothesized that LPS-HI would acutely deplete reduced glutathione (GSH), indicating overwhelming oxidative stress in spite of hypothermia treatment in neonatal rats. Post-natal day 7 rats were randomized to sham ligation, or severe LPS-HI (0.5 mg/kg 4 h before right carotid artery ligation, 90 min 8% O2), followed by hypothermia alone or with N-acetylcysteine (25 mg/kg) and vitamin D (1,25(OH)2D3, 0.05 μg/kg) (NVD). We quantified in vivo CNS metabolites by serial 7T MR Spectroscopy before, immediately after LPS-HI, and after treatment, along with terminal plasma drug concentrations. GSH was significantly decreased in all LPS-HI rats compared with baseline and sham controls. Two hours of hypothermia alone did not improve GSH and allowed glutamate + glutamine (GLX) to increase. Within 1 h of administration, NVD increased GSH close to baseline and suppressed GLX. The combination of NVD with hypothermia rapidly improved cellular redox status after LPS-HI, potentially inhibiting important secondary injury cascades and allowing more time for hypothermic neuroprotection.

PURPOSE: Diffusion tensor imaging (DTI) can detect injury to specific white matter (WM) tracts involved with sensorimotor processing and may provide sensitive measures for latent or nascent motor skills. We hypothesized that DTI measures of WM fractional anisotropy (FA) could predict early motor scores on a standardized assessment in a cohort of preterm infants at risk for WM injury. METHODS: In this prospective study, preterm infants (n = 26, 11 female, 15 male, mean gestational age 29.1 ± 2.5) underwent the Test of Infant Motor Performance (TIMP) at term and at 12 weeks corrected age (CA) and underwent an non-sedated magnetic resonance imaging (MRI) with DTI at a mean of 42 ± 1.5 weeks CA. Fractional anisotropy (FA) was measured by Voxelwise statistical analysis using Tract-Based Spatial Statistics (TBSS) in the specific regions of interest. RESULTS: Significant differences were found between infants with poor versus average performance on motor assessments at 12-weeks and FA values in several left hemispheric WM tracts ( p< 0.05). High FA of the left anterior limb of the internal capsule (ALIC) predicted mean increase in TIMP scores on specific items for head lift in prone and head lift turn to sound (p = 0.045 and p = 0.002). CONCLUSION: Subtle WM injury, as indicated by low FA in left WM tracts, can predict outcomes of early motor skills performance testing at 3 months. Early DTI may identify infants with silent WM injury who need early intervention. Further studies may establish if individual tract FA improve after targeted treatment.

PURPOSE This study used kinematic analysis to identify a reliable and rapid assessment method for abnormal patterns of motor development in preterm infants. METHODS In a retrospective analysis, we examined video of n= 35 preterm infants at 3mo corrected age (CA) who had concurrent Test of Infant Motor Performance (TIMP) scores. Hyperflexion at the hip produces common gait anomalies seen in children with CP, therefore we analyzed hip angle in the prone head lift position at 3 months CA. Magnetic Resonance Spectroscopy (MRS) was performed at term equivalent (n= 23) and Bayley-III neurodevelopmental tests were performed at 1 year (n= 28). We correlated hip angles with TIMP and Bayley-III scores, and MRS neuronal metabolites. RESULTS Hip angle positively correlated with TIMP at 3 months (r= 0.642, p≤ 0.001), but not with Bayley-III at 1 year (r= 0.122, p= 0.529). Hip angle correlated negatively with myo-inositol (mI) ratios in frontal white matter tracts (mI/Cr r= -0.520, p= 0.011). CONCLUSIONS These results suggest prone hip angle may be a quantitative proxy for the 42-item TIMP at 3 months, and that hypertonicity in the hip flexor musculature is a manifestation of white matter metabolic abnormalities (elevated mI ratios) that may indicate occult white matter injury.

Background Proton magnetic resonance spectroscopy (1H-MRS) studies have consistently found abnormal brain concentrations of N-acetylaspartate (NAA) and glutamate in individuals with alcohol use disorders (AUD) relative to light drinkers. However, most such studies have focused on individuals in treatment for severe alcohol dependence (AD), and few studies have investigated associations between neurochemical concentrations and recent alcohol consumption. This study focused on associations between recent drinking and prefrontal neurometabolite concentrations in nonsevere, non-treatment-seeking individuals with AUD. Methods Nineteen treatment-naive alcohol-dependent individuals aged 21 to 40 completed a 1H-MRS scan. Single-voxel 1H-MRS spectra were acquired in dorsal anterior cingulate cortex (dACC) using a 2-dimensional J-resolved point resolved spectroscopy sequence. Associations between recent heavy drinking, assessed using the Timeline FollowBack, and dACC metabolite concentrations were estimated via regression controlling for within-voxel tissue composition. Results Participants provided a negative breathalyzer reading and reported between 1 and 5 days (M = 2.45, SD = 1.23) since their last drink. Number of heavy drinking days in the 14 days preceding the scan (M = 4.84, SD = 3.32) was significantly inversely associated with both glutamate/water (β = −0.63, t(17) = −3.37, p = 0.004) and NAA/water concentrations (β = −0.59, t(17) = −2.98, p = 0.008). Conclusions This study extends the literature by demonstrating inverse associations between recent heavy drinking and dACC glutamate and NAA concentrations in a sample of nonsevere, non-treatment-seeking individuals with AD. These findings may support the hypothesis that amount of recent alcohol consumption may account for differences in neuronal metabolism, even in nonsevere, non-treatment-seeking alcoholics.

Persistent oxidative stress depletes reduced glutathione (GSH), an intracellular antioxidant and an important determinant of CNS injury after hypoxia ischemia. We used standard, short echo time Stimulated Echo Acquisition Mode (STEAM) to detect GSH by magnetic resonance spectroscopy (MRS) in 24 term neonates with hypoxic-ischemic encephalopathy (HIE), on day of life 5–6, after rewarming from therapeutic hypothermia. MRS demonstrated reliable, consistent GSH of 1·64 ± 0·20 mM in the basal ganglia immediately before intravenous infusion of N-acetylcysteine. N-acetylcysteine resulted in a rapid and significant GSH increase to 1·93 ± 0.23 mM within 12–30 min after completion of infusion ( n = 21, p