EEG slow waves in traumatic brain injury: Convergent findings in mouse and man.

OBJECTIVE: Evidence from previous studies suggests that greater sleep pressure, in the form of EEG-based slow waves, accumulates in specific brain regions that are more active during prior waking experience. We sought to quantify the number and coherence of EEG slow waves in subjects with mild traumatic brain injury (mTBI). METHODS: We developed a method to automatically detect individual slow waves in each EEG channel, and validated this method using simulated EEG data. We then used this method to quantify EEG-based slow waves during sleep and wake states in both mouse and human subjects with mTBI. A modified coherence index that accounts for information from multiple channels was calculated as a measure of slow wave synchrony. RESULTS: Brain-injured mice showed significantly higher theta:alpha amplitude ratios and significantly more slow waves during spontaneous wakefulness and during prolonged sleep deprivation, compared to sham-injured control mice. Human subjects with mTBI showed significantly higher theta:beta amplitude ratios and significantly more EEG slow waves while awake compared to age-matched control subjects. We then quantified the global coherence index of slow waves across several EEG channels in human subjects. Individuals with mTBI showed significantly less EEG global coherence compared to control subjects while awake, but not during sleep. EEG global coherence was significantly correlated with severity of post-concussive symptoms (as assessed by the Neurobehavioral Symptom Inventory scale). CONCLUSION AND IMPLICATIONS: Taken together, our data from both mouse and human studies suggest that EEG slow wave quantity and the global coherence index of slow waves may represent a sensitive marker for the diagnosis and prognosis of mTBI and post-concussive symptoms.

EEG slow waves in traumatic brain injury: Convergent findings in mouse and man.

OBJECTIVE: Evidence from previous studies suggests that greater sleep pressure, in the form of EEG-based slow waves, accumulates in specific brain regions that are more active during prior waking experience. We sought to quantify the number and coherence of EEG slow waves in subjects with mild traumatic brain injury (mTBI). METHODS: We developed a method to automatically detect individual slow waves in each EEG channel, and validated this method using simulated EEG data. We then used this method to quantify EEG-based slow waves during sleep and wake states in both mouse and human subjects with mTBI. A modified coherence index that accounts for information from multiple channels was calculated as a measure of slow wave synchrony. RESULTS: Brain-injured mice showed significantly higher theta:alpha amplitude ratios and significantly more slow waves during spontaneous wakefulness and during prolonged sleep deprivation, compared to sham-injured control mice. Human subjects with mTBI showed significantly higher theta:beta amplitude ratios and significantly more EEG slow waves while awake compared to age-matched control subjects. We then quantified the global coherence index of slow waves across several EEG channels in human subjects. Individuals with mTBI showed significantly less EEG global coherence compared to control subjects while awake, but not during sleep. EEG global coherence was significantly correlated with severity of post-concussive symptoms (as assessed by the Neurobehavioral Symptom Inventory scale). CONCLUSION AND IMPLICATIONS: Taken together, our data from both mouse and human studies suggest that EEG slow wave quantity and the global coherence index of slow waves may represent a sensitive marker for the diagnosis and prognosis of mTBI and post-concussive symptoms.

Low-temperature dynamic nuclear polarization of gases in Frozen mixtures.

PURPOSE: To present a new cryogenic technique for preparing gaseous compounds in solid mixtures for polarization using dynamic nuclear polarization (DNP). METHODS: (129) Xe and (15) N2 O samples were prepared using the presented method. Samples were hyperpolarized at 1.42K at 5 Tesla. (129) Xe was polarized at 1.65K and 1.42K to compare enhancement. Polarization levels for both samples and T1 relaxation times for the (129) Xe sample were measured. Sample pulverization for the (129) Xe and controlled annealing for both samples were introduced as additional steps in sample preparation. RESULTS: Enhancement increased by 15% due to a temperature drop from 1.65K to 1.42K for the (129) Xe sample. A polarization level of 20 ± 3% for the (129) Xe sample was achieved, a two-fold increase from 10 ± 1% after pulverization of the sample at 1.42K. T1 of the (129) Xe sample was increased by more than three-fold by means of annealing. In the case of (15) N2 O, annealing led to a ∼two-fold increase in the signal level after DNP. CONCLUSION: The presented technique for producing and manipulating solid gas/glassing agent/radical mixtures for DNP led to high polarization levels in (129) Xe and (15) N2 O samples. These methods show potential for polarizing other gases using DNP technology. Magn Reson Med 76:1007-1014, 2016. © 2015 Wiley Periodicals, Inc.

Versatile pulse sequence device to conserve hyperpolarization for NMR and MRI studies.

PURPOSE: Levitt and co-workers have described the M2S pulse sequence which transfers between longitudinal and singlet spin order. Building on this work, we describe the construction of a portable M2S pulse sequence generator to increase the relaxation time of polarized compounds. Additionally, we investigate the efficiency of spin order transfer under conditions where physical parameters of the system are not known precisely. THEORY AND METHODS: A portable M2S generator is built. Longitudinally polarized N2O is converted to the singlet state by both adiabatic transfer and by the M2S sequence. Density matrix simulations are used to model the effects of mismatched chemical shift, flip angle, and scalar couplings. RESULTS: Density matrix simulations suggest that to convert 95% of the longitudinal m = 1 triplet state population to the singlet order we must match the Larmor precession frequency to the excitation radiofrequency field by 10%, the scalar couplings must be determined to better than 0.6%, and the flip angle must be calibrated to better than 2%. CONCLUSION: The sequence is robust against many mismatched physical parameters of the species we are converting. Additionally, the instrument's portability allows for the conversion of hyperpolarized species near a polarizer. The lifetime is increased by ∼12-fold. This is highly advantageous in systems where the hyperpolarized media relax rapidly.

Dietary therapy mitigates persistent wake deficits caused by mild traumatic brain injury.

Sleep disorders are highly prevalent in patients with traumatic brain injury (TBI) and can significantly impair cognitive rehabilitation. No proven therapies exist to mitigate the neurocognitive consequences of TBI. We show that mild brain injury in mice causes a persistent inability to maintain wakefulness and decreases orexin neuron activation during wakefulness. We gave mice a dietary supplement of branched-chain amino acids (BCAAs), precursors for de novo glutamate synthesis in the brain. BCAA therapy reinstated activation of orexin neurons and improved wake deficits in mice with mild brain injury. Our data suggest that dietary BCAA intervention, acting in part through orexin, can ameliorate injury-induced sleep disturbances and may facilitate cognitive rehabilitation after brain injury.

Positive end-expiratory pressure increments during anesthesia in normal lung result in hysteresis and greater numbers of smaller aerated airspaces.

BACKGROUND: Although it is recognized that pulmonary hysteresis can influence the effects of positive end-expiratory pressure (PEEP), the extent to which expansion of previously opened (vs. newly opening) peripheral airspaces contribute to increased lung volume is unknown. METHODS: Following a recruitment maneuver, rats were ventilated with constant tidal volumes and imaged during ascending and descending ramps of PEEP. RESULTS: The authors estimated peripheral airspace dimensions by measuring the apparent diffusion coefficient of He in 10 rats. In a separate group (n = 5) undergoing a similar protocol, the authors used computerized tomography to quantify lung volume. Hysteresis was confirmed by larger end-inspiratory lung volume (mean ± SD; all PEEP levels included): 8.4 ± 2.8 versus 6.8 ± 2.0 ml (P < 0.001) and dynamic compliance: 0.52 ± 0.12 versus 0.42 ± 0.09 ml/cm H2O (P < 0.001) during descending versus ascending PEEP ramps. Apparent diffusion coefficient increased with PEEP, but it was smaller during the descending versus ascending ramps for corresponding levels of PEEP: 0.168 ± 0.019 versus 0.183 ± 0.019 cm/s (P < 0.001). Apparent diffusion coefficient was smaller in the posterior versus anterior lung regions, but the effect of PEEP and hysteresis on apparent diffusion coefficient was greater in the posterior regions. CONCLUSIONS: The authors' study results suggest that in healthy lungs, larger lung volumes due to hysteresis are associated with smaller individual airspaces. This may be explained by opening of previously nonaerated peripheral airspaces rather than expansion of those already aerated. Setting PEEP on a descending ramp may minimize distension of individual airspaces.

A variability study of regional alveolar oxygen tension measurement in humans using hyperpolarized (3) He MRI.

PURPOSE: A systematic study of the short-term and long-term variability of regional alveolar partial pressure of oxygen tension (pA O2 ) measurements using (3) He magnetic resonance imaging was presented. Additionally, the repeatability of the average evaluated pA O2 was compared with that of the standard pulmonary function tests. METHODS: Pulmonary function test and pA O2 imaging were performed on 4 nonsmokers (1 M, 3 F, 56 ± 1.7 years) and 4 smokers (3 M, 1 F, 52 ± 7.5 years) during three visits over the course of 2 weeks. Two measurements were performed per visit. Variability of pA O2 was assessed using a mixed-effect model, with an intraclass correlation coefficient calculated for each group. The coefficient of variation of pA O2 over the 3-day period was also compared with the coefficient of variation of pulmonary function test results. RESULTS: Short-term regional variability based on intraclass correlation coefficient was 0.71 for nonsmokers, and 0.63 for smokers, with long-term variability significantly lower at 0.59 and 0.47, respectively. While the coefficient of variation of the average pA O2 was similar to the repeatability of the diffusing capacity of CO, it was significantly higher than that of Forced Vital Capacity (P = 0.02). CONCLUSION: Short-term and long-term pA O2 variability differences were used as an indication of true physiological changes in order to measure technical reproducibility. Smokers show higher physiologic variability and less technical reproducibility. The suggested pA O2 -imaging technique showed a reasonable regional repeatability in nonsmokers as well as the ability to detect differences between the two groups with similar reproducibility and superior discriminatory ability when compared with pulmonary function tests.

Imaging the interaction of atelectasis and overdistension in surfactant-depleted lungs.

OBJECTIVE: Atelectasis and surfactant depletion may contribute to greater distension-and thereby injury-of aerated lung regions; recruitment of atelectatic lung may protect these regions by attenuating such overdistension. However, the effects of atelectasis (and recruitment) on aerated airspaces remain elusive. We tested the hypothesis that during mechanical ventilation, surfactant depletion increases the dimensions of aerated airspaces and that lung recruitment reverses these changes. DESIGN: Prospective imaging study in an animal model. SETTING: Research imaging facility. SUBJECTS: Twenty-seven healthy Sprague Dawley rats. INTERVENTIONS: Surfactant depletion was obtained by saline lavage in anesthetized, ventilated rats. Alveolar recruitment was accomplished using positive end-expiratory pressure and exogenous surfactant administration. MEASUREMENTS AND MAIN RESULTS: Airspace dimensions were estimated by measuring the apparent diffusion coefficient of He, using diffusion-weighted hyperpolarized gas magnetic resonance imaging. Atelectasis was demonstrated using computerized tomography and by measuring oxygenation. Saline lavage increased atelectasis (increase in nonaerated tissue from 1.2% to 13.8% of imaged area, p < 0.001), and produced a concomitant increase in mean apparent diffusion coefficient (~33%, p < 0.001) vs. baseline; the heterogeneity of the computerized tomography signal and the variance of apparent diffusion coefficient were also increased. Application of positive end-expiratory pressure and surfactant reduced the mean apparent diffusion coefficient (~23%, p < 0.001), and its variance, in parallel to alveolar recruitment (i.e., less computerized tomography densities and heterogeneity, increased oxygenation). CONCLUSIONS: Overdistension of aerated lung occurs during atelectasis is detectable using clinically relevant magnetic resonance imaging technology, and could be a key factor in the generation of lung injury during mechanical ventilation. Lung recruitment by higher positive end-expiratory pressure and surfactant administration reduces airspace distension.

Determination of the singlet state lifetime of dissolved nitrous oxide from high field relaxation measurements.

Longitudinal spin relaxation due to modulation of dipolar interactions often limits the development of hyperpolarized magnetic tracers. Recently, it has been demonstrated that transferring spin order to a singlet state significantly increases the polarization lifetimes in systems where nitrous oxide is dissolved in a liquid solvent. Additionally, previous studies have suggested that the longitudinal relaxation of nitrous oxide is largely dominated by the spin-rotation interaction. Models of spin-relaxation under Brownian motion naïvely predict the angular momentum reorienting correlation time of the spin rotation interaction to be inversely proportional to the viscosity of the solution. This dependence implies the singlet lifetime can be lengthened by increasing the dissolving solvent's viscosity-an extension which is not observed. Our work formulates a model which describes the relaxation of nitrous oxide dissolved in various solvents. We investigate the effect of altering the temperature of the solvent, as well as the effect of varying solute-solvent interactions on the singlet state as well as the longitudinal polarization lifetime. We predict the singlet lifetime for nitrous oxide dissolved in several solvents by fitting rotational and angular momentum correlation times measured at high magnetic field, and relate singlet relaxation to translational diffusion constants.

A multislice single breath-hold scheme for imaging alveolar oxygen tension in humans.

Reliable, noninvasive, and high-resolution imaging of alveolar partial pressure of oxygen (p(A)O(2)) is a potentially valuable tool in the early diagnosis of pulmonary diseases. Several techniques have been proposed for regional measurement of p(A)O(2) based on the increased depolarization rate of hyperpolarized (3) He. In this study, we explore one such technique by applying a multislice p(A)O(2) -imaging scheme that uses interleaved-slice ordering to utilize interslice time-delays more efficiently. This approach addresses the low spatial resolution and long breath-hold requirements of earlier techniques, allowing p(A)O(2) measurements to be made over the entire human lung in 10-15 s with a typical resolution of 8.3 × 8.3 × 15.6 mm(3). PO(2) measurements in a glass syringe phantom were in agreement with independent gas analysis within 4.7 ± 4.1% (R = 0.9993). The technique is demonstrated in four human subjects (healthy nonsmoker, healthy former smoker, healthy smoker, and patient with COPD), each imaged six times on 3 different days during a 2-week span. Two independent measurements were performed in each session, consisting of 12 coronal slices. The overall p(A)O(2) mean across all subjects was 95.9 ± 12.2 Torr and correlated well with end-tidal O(2) (R = 0.805, P < 0.0001). The alveolar O(2) uptake rate was consistent with the expected range of 1-2 Torr/s. Repeatable visual features were observed in p(A)O(2) maps over different days, as were characteristic differences among the subjects and gravity-dependent effects.

Cryptophane xenon-129 nuclear magnetic resonance biosensors targeting human carbonic anhydrase.

(129)Xe NMR biosensors are promising agents for early disease detection, especially when their interactions with target biomolecules can perturb (129)Xe chemical shifts well beyond the typical field inhomogeneity of clinical MRI. We introduce human carbonic anhydrase (CA) as a single-binding-site enzyme for studying xenon biosensor-protein interactions. A xenon-binding cryptophane was substituted with linkers of varying lengths to p-benzenesulfonamide to yield nondiastereomeric biosensors with a single (129)Xe NMR resonance. X-ray crystallography confirmed binding of the eight-bond-linked biosensor containing a single xenon atom in the CAII active site. Biosensor dissociation constants (K(d) = 20-110 nM) were determined by isothermal titration calorimetry (ITC) for isozymes CA I and II. The biosensor-CA complexes yielded "bound" hyperpolarized (129)Xe NMR resonances of narrow line width that were shifted by 3.0-7.5 ppm downfield, signifying much larger shifts than seen previously. Moreover, isozyme-specific chemical shifts clearly differentiated CA I and II, despite their similar structures. Thus, xenon biosensors may provide a powerful strategy for diagnosing human diseases characterized by the upregulation of specific CA isozymes and other protein biomarkers.

Designing 129Xe NMR biosensors for matrix metalloproteinase detection.

Xenon-129 biosensors offer an attractive alternative to conventional MRI contrast agents due to the chemical shift sensitivity and large nuclear magnetic signal of hyperpolarized (129)Xe. Here, we report the first enzyme-responsive (129)Xe NMR biosensor. This compound was synthesized in 13 steps by attaching the consensus peptide substrate for matrix metalloproteinase-7 (MMP-7), an enzyme that is upregulated in many cancers, to the xenon-binding organic cage, cryptophane-A. The final coupling step was achieved on solid support in 80-92% yield via a copper (I)-catalyzed [3+2] cycloaddition. In vitro enzymatic cleavage assays were monitored by HPLC and fluorescence spectroscopy. The biosensor was determined to be an excellent substrate for MMP-7 (K(M) = 43 microM, V(max) = 1.3 x 10(-)(8) M s(-1), k(cat)/K(M) = 7,200 M(-1) s(-1)). Enzymatic cleavage of the tryptophan-containing peptide led to a dramatic decrease in Trp fluorescence, lambda(max) = 358 nm. Stern-Volmer analysis gave an association constant of 9000 +/- 1,000 M(-1) at 298 K between the cage and Trp-containing hexapeptide under enzymatic assay conditions. Most promisingly, (129)Xe NMR spectroscopy distinguished between the intact and cleaved biosensors with a 0.5 ppm difference in chemical shift. This difference most likely reflected a change in the electrostatic environment of (129)Xe, caused by the cleavage of three positively charged residues from the C-terminus. This work provides guidelines for the design and application of new enzyme-responsive (129)Xe NMR biosensors.