Quantitative and simultaneous measurement of oxygen consumption rates in rat brain and skeletal muscle using 17 O MRS imaging at 16.4T.

PURPOSE: Oxygen-17 (17 O) MRS imaging, successfully used in the brain, is extended by imaging the oxygen metabolic rate in the resting skeletal muscle and used to determine the total whole-body oxygen metabolic rate in the rat. METHODS: During and after inhalations of 17 O2 gas, dynamic 17 O MRSI was performed in rats (n = 8) ventilated with N2 O or N2 at 16.4 T. Time courses of the H217 O concentration from regions of interest located in brain and muscle tissue were examined and used to fit an animal-adapted 3-phase metabolic model of oxygen consumption. CBF was determined with an independent washout method. Finally, body oxygen metabolic rate was calculated using a global steady-state approach. RESULTS: Cerebral metabolic rate of oxygen consumption was 1.97 ± 0.19 µmol/g/min on average. The resting metabolic rate of oxygen consumption in skeletal muscle was 0.32 ± 0.12 µmol/g/min and >6 times lower than cerebral metabolic rate of oxygen consumption. Global oxygen consumed by the body was 24.2 ± 3.6 mL O2 /kg body weight/min. CBF was estimated to be 0.28 ± 0.02 mL/g/min and 0.34 ± 0.06 mL/g/min for the N2 and N2 O ventilation condition, respectively. CONCLUSION: We have evaluated the feasibility of 17 O MRSI for imaging and quantifying the oxygen consumption rate in low metabolizing organs such as the skeletal muscle at rest. Additionally, we have shown that CBF is slightly increased in the case of ventilation with N2 O. We expect this study to be beneficial to the application of 17 O MRSI to a wider range of organs, although further validation is advised.

Ultra-High Field MRI in Alzheimer's Disease: Effective Transverse Relaxation Rate and Quantitative Susceptibility Mapping of Human Brain In Vivo and Ex Vivo compared to Histology.

Alzheimer's disease (AD) is the most common cause of dementia worldwide. So far, diagnosis of AD is only unequivocally defined through postmortem histology. Amyloid plaques are a classical hallmark of AD and amyloid load is currently quantified by Positron Emission tomography (PET) in vivo. Ultra-high field magnetic resonance imaging (UHF-MRI) can potentially provide a non-invasive biomarker for AD by allowing imaging of pathological processes at a very-high spatial resolution. The first aim of this work was to reproduce the characteristic cortical pattern previously observed in vivo in AD patients using weighted-imaging at 7T. We extended these findings using quantitative susceptibility mapping (QSM) and quantification of the effective transverse relaxation rate (R2*) at 9.4T. The second aim was to investigate the origin of the contrast patterns observed in vivo in the cortex of AD patients at 9.4T by comparing quantitative UHF-MRI (9.4T and 14.1T) of postmortem samples with histology. We observed a distinctive cortical pattern in vivo in patients compared to healthy controls (HC), and these findings were confirmed ex vivo. Specifically, we found a close link between the signal changes detected by QSM in the AD sample at 14.1T and the distribution pattern of amyloid plaques in the histological sections of the same specimen. Our findings showed that QSM and R2* maps can distinguish AD from HC at UHF by detecting cortical alterations directly related to amyloid plaques in AD patients. Furthermore, we provided a method to quantify amyloid plaque load in AD patients at UHF non-invasively.

Widespread and Opponent fMRI Signals Represent Sound Location in Macaque Auditory Cortex.

In primates, posterior auditory cortical areas are thought to be part of a dorsal auditory pathway that processes spatial information. But how posterior (and other) auditory areas represent acoustic space remains a matter of debate. Here we provide new evidence based on functional magnetic resonance imaging (fMRI) of the macaque indicating that space is predominantly represented by a distributed hemifield code rather than by a local spatial topography. Hemifield tuning in cortical and subcortical regions emerges from an opponent hemispheric pattern of activation and deactivation that depends on the availability of interaural delay cues. Importantly, these opponent signals allow responses in posterior regions to segregate space similarly to a hemifield code representation. Taken together, our results reconcile seemingly contradictory views by showing that the representation of space follows closely a hemifield code and suggest that enhanced posterior-dorsal spatial specificity in primates might emerge from this form of coding.

Assessing White Matter Microstructure in Brain Regions with Different Myelin Architecture Using MRI.

OBJECTIVE: We investigate how known differences in myelin architecture between regions along the cortico-spinal tract and frontal white matter (WM) in 19 healthy adolescents are reflected in several quantitative MRI parameters that have been proposed to non-invasively probe WM microstructure. In a clinically feasible scan time, both conventional imaging sequences as well as microstructural MRI parameters were assessed in order to quantitatively characterise WM regions that are known to differ in the thickness of their myelin sheaths, and in the presence of crossing or parallel fibre organisation. RESULTS: We found that diffusion imaging, MR spectroscopy (MRS), myelin water fraction (MWF), Magnetization Transfer Imaging, and Quantitative Susceptibility Mapping were myelin-sensitive in different ways, giving complementary information for characterising WM microstructure with different underlying fibre architecture. From the diffusion parameters, neurite density (NODDI) was found to be more sensitive than fractional anisotropy (FA), underlining the limitation of FA in WM crossing fibre regions. In terms of sensitivity to different myelin content, we found that MWF, the mean diffusivity and chemical-shift imaging based MRS yielded the best discrimination between areas. CONCLUSION: Multimodal assessment of WM microstructure was possible within clinically feasible scan times using a broad combination of quantitative microstructural MRI sequences. By assessing new microstructural WM parameters we were able to provide normative data and discuss their interpretation in regions with different myelin architecture, as well as their possible application as biomarker for WM disorders.

(17)O relaxation times in the rat brain at 16.4 tesla.

PURPOSE: Measurement of the cerebral metabolic rate of oxygen (CMRO2 ) by means of direct imaging of the (17) O signal can be a valuable tool in neuroscientific research. However, knowledge of the longitudinal and transverse relaxation times of different brain tissue types is required, which is difficult to obtain because of the low sensitivity of natural abundance H2 (17) O measurements. METHODS: Using the improved sensitivity at a field strength of 16.4 Tesla, relaxation time measurements in the rat brain were performed in vivo and postmortem with relatively high spatial resolutions, using a chemical shift imaging sequence. RESULTS: In vivo relaxation times of rat brain were found to be T1 = 6.84 ± 0.67 ms and T2 * = 1.77 ± 0.04 ms. Postmortem H2 (17) O relaxometry at enriched concentrations after inhalation of (17) O2 showed similar T2 * values for gray matter (1.87 ± 0.04 ms) and white matter, significantly longer than muscle (1.27 ± 0.05 ms) and shorter than cerebrospinal fluid (2.30 ± 0.16 ms). CONCLUSION: Relaxation times of brain H2 (17) O were measured for the first time in vivo in different types of tissues with high spatial resolution. Because the relaxation times of H2 (17) O are expected to be independent of field strength, our results should help in optimizing the acquisition parameters for experiments also at other MRI field strengths.

Functional quantitative susceptibility mapping (fQSM).

Blood oxygenation level dependent (BOLD) functional magnetic resonance imaging (fMRI) is a powerful technique, typically based on the statistical analysis of the magnitude component of the complex time-series. Here, we additionally interrogated the phase data of the fMRI time-series and used quantitative susceptibility mapping (QSM) in order to investigate the potential of functional QSM (fQSM) relative to standard magnitude BOLD fMRI. High spatial resolution data (1mm isotropic) were acquired every 3 seconds using zoomed multi-slice gradient-echo EPI collected at 7 T in single orientation (SO) and multiple orientation (MO) experiments, the latter involving 4 repetitions with the subject's head rotated relative to B0. Statistical parametric maps (SPM) were reconstructed for magnitude, phase and QSM time-series and each was subjected to detailed analysis. Several fQSM pipelines were evaluated and compared based on the relative number of voxels that were coincidentally found to be significant in QSM and magnitude SPMs (common voxels). We found that sensitivity and spatial reliability of fQSM relative to the magnitude data depended strongly on the arbitrary significance threshold defining "activated" voxels in SPMs, and on the efficiency of spatio-temporal filtering of the phase time-series. Sensitivity and spatial reliability depended slightly on whether MO or SO fQSM was performed and on the QSM calculation approach used for SO data. Our results present the potential of fQSM as a quantitative method of mapping BOLD changes. We also critically discuss the technical challenges and issues linked to this intriguing new technique.

Monitoring the stress-level of rats with different types of anesthesia: a tail-artery cannulation protocol.

INTRODUCTION: Functional MRI in rats under anesthesia can largely minimize motion artifacts and attenuate the stress of the animal. However, two issues remain to be clarified and improved. First, fMRI results obtained with different types of anesthesia during surgical preparation and imaging show a large variability, which could be caused by the variable stress level of the rodents. Second, the most common surgical procedure used for anesthesia, blood gas analysis and mean arterial blood-pressure (MABP) monitoring is the femoral vein and artery catheterization that makes longitudinal studies difficult. METHODS: In order to examine the variability of the stress level with three different anesthesia protocols using isoflurane (Iso), medetomidine-ketamine (MK) or propofol-remifentanil (PR), we measured the plasma corticosterone (CORT) concentration with (125)I-radioimmunoassay in blood samples collected prior to, immediately after and 60min after surgery. Tail-artery and vein catheterization was adapted for long-term monitoring of MABP with periodic blood sampling and is proposed as a less invasive and technically simple alternative to femoral vessel catheterization in fMRI preparation protocols. RESULTS: We show that the CORT concentration depends on the anesthesia protocol with both alternatives providing more efficient stress reduction than the protocol using Iso. However, only the protocol using PR achieved a significant hormone reduction during surgery. Stress was not reliably manifested in changes in heart-rate and breathing-rate. Anesthesia and strain related changes in these two physiological parameters may be assigned to the pharmacological effects of the premedication and anesthetic agents. The results indicate also that MABP can be monitored over a long period of time (e.g. functional imaging session) through an arterial access point in the rat tail after cannulation with the proposed procedure. DISCUSSION AND CONCLUSION: Animals can experience stress during fMRI preparation protocols without obvious signs in commonly monitored physiological parameters. Our results challenge the efficiency of surgical protocols using Iso as mono-anesthetic agent, even when extended with topical analgesia. It was demonstrated that the CORT-based stress-level measurement through tail-artery cannulation can be used for developing anesthesia protocols (i.e. the presented PR protocol) when setting up future fMRI studies. The proposed surgical method for the tail is expected to facilitate longitudinal fMRI studies with permanent arterial access.

In vivo visualization of single native pancreatic islets in the mouse.

The purpose of this study was to investigate the potential of a novel targeted contrast agent (CA) for the in vivo visualization of single native pancreatic islets, the sites of insulin production, in the pancreas of mice using magnetic resonance imaging (MRI). The CA for intravenous administration was composed of the ß-cell-specific single-chain antibody fragment, SCA B1, and ferromagnetic carbon-coated cobalt nanoparticles. MRI experiments were performed at 7, 9.4 and 16.4 T in excised organs (pancreas, liver, kidney, spleen), at 7 T in mice fixed in formalin and at 9.4 and 16.4 T in living mice. Image contrast in untreated control animals was compared with images from mice treated with unspecific and specific CA. For the validation of MRI results, selected pancreases were subjected to immunohistochemical staining and numerical contrast simulations were performed. Ex vivo results and the outcome of immunohistochemistry suggest that islets are marked only by the CA containing SCA B1. Strong accumulation of particles was found also in other investigated organs owing to the uptake by the reticuloendothelial system, but the contrast in the MR images is clearly distinguishable from the islet specific contrast in pancreases and numerical predictions. In vivo experiments based on averaged dynamic sampling with 66 × 66 × 100 µm³ and triggered acquisition with 90 × 90 × 200 µm³ nominal resolution resulted in similar particle contrast to in in vitro measurements. The newly developed CA and MRI strategies have the potential to be used for studying mouse diabetes models by visualizing single native pancreatic islets.

Rat brain MRI at 16.4T using a capacitively tunable patch antenna in combination with a receive array.

For MRI at 16.4T, with a proton Larmor frequency of 698 MHz, one of the principal RF engineering challenges is to generate a spatially homogeneous transmit field over a larger volume of interest for spin excitation. Constructing volume coils large enough to house a receive array along with the subject and to maintain the quadrature symmetry for different loading conditions is difficult at this frequency. This calls for new approaches to RF coil design for ultra-high field MR systems. A remotely placed capacitively tunable patch antenna, which can easily be adjusted to different loading conditions, was used to generate a relatively homogeneous excitation field covering a large imaging volume with a transversal profile similar to that of a birdcage coil. Since it was placed in front of the animal, this created valuable free space in the narrow magnet bore around the subject for additional hardware. To enhance the reception sensitivity, the patch antenna was combined with an actively detunable 3-channel receive coil array. In addition to increased SNR compared to a quadrature transceive surface coil, we were able to get high quality gradient echo and spin-echo images covering the whole rat brain.

Determination of regional variations and reproducibility in in vivo 1H NMR spectroscopy of the rat brain at 16.4 T.

In vivo (1)H NMR spectroscopy was used to obtain the neurochemical profile in the posterior parts of the brain, the cerebellum and the medulla oblongata in comparison to the hippocampus and the thalamus. Using small voxel sizes between 16 and 32 µl to avoid partial volume effects, most metabolites demonstrated significant regional differences except acetate, γ-aminobutyric acid, and phosphorylcholine. Noticeable regional differences in metabolite concentrations were the significant increase of total creatine in the cerebellum and the substantial decrease of taurine in thalamus and medulla oblongata. In particular, the glycine concentration in the medulla oblongata was determined to be 4.37 ± 0.68 µmol/g (Cramér-Rao lower bounds 7%) and thus significantly higher than in the other regions, consistent with findings reported in both in vivo (1)H NMR spectroscopy and in vitro biochemical assays. Intraindividual reproducibility and interindividual variability were investigated by acquiring spectra from the thalamus of the same rats in two sessions. No prominent influence on measurement session was observed in metabolite concentrations with coefficients of variations being below 20% in 16 metabolites.

Rat strain-dependent variations in brain metabolites detected by in vivo (1) H NMR spectroscopy at 16.4T.

Localized in vivo (1) H NMR spectroscopy is playing an increasing role in preclinical studies, because of its ability to quantify the concentrations of up to 20 metabolites in rat brain. To assess the differences between often-used rat strains, the neurochemical profiles of Sprague-Dawley, Wistar and Fischer rats were determined at ultrashort TE at 16.4 T. To ascertain high-qualitative quantification, a first experiment examined the dependence of the measuring time on the quantification results and precision by precisely the number of averages between 16 and 320. It was shown that most metabolites can be quantified accurately within a short scan time, yielding Cramér-Rao lower bounds below 20% and stable concentrations for 16 metabolites with as few as 32 or 64 averages in the thalamus and hippocampus, respectively. Interstrain differences in metabolite concentrations were shown to be moderate, with taurine varying significantly between Sprague-Dawley and Wistar rats, and slightly more distinct differences from Fischer rats, including variations in glutamate and myo-inositol. The high spectral quality and quantification precision of all data again demonstrated the potential of in vivo (1)H NMR spectroscopy at ultrahigh field.

Intermolecular zero-quantum coherence NMR spectroscopy in the presence of local dipole fields.

NMR experiments detecting intermolecular zero-quantum coherences (iZQCs) allow for observation of homogeneous line shapes under inhomogeneous magnetic fields. Local dipole fields impair the refocusing capacity of such experiments and render the available theoretical description of signal evolution invalid. In this article, the impact of local dipole fields on two-dimensional iZQC spectroscopy experiments was assessed by performing extensive numerical simulations, which solved the nonlinear Bloch equations for a binary solution in a magnetization array of 64(3) spatial points. Local dipole fields were simulated using spherical volumes with different magnetic susceptibility values corresponding to either a glass sphere or an air inclusion with a diameter of 100 microm. The local field resulted in a broadened distribution of difference frequencies between locally interacting spins and led to the dominating effect of decreasing the amplitude of the solute peak, before line broadening was observed in the spectra. From simulations using a magnetic field strength of 17.6 T, the smallest ratio of sample to inclusion volume that still allowed for observation of the solute peak was determined to be eta(limit)=215 and eta(limit)=392 for glass and air inclusions, respectively. Experimental data acquired with a 100 microm diameter glass sphere embedded in agar gel yielded a value of eta(limit)=252 and confirmed the order of magnitude obtained from the simulations. From these data, it was concluded that iZQC spectroscopy is possible as long as the relative volume occupied by air inclusions does not exceed the order of 0.1% of the sample volume. This limit, in contrast to the previous speculations, strongly excludes materials or tissues with high density of strong inhomogeneities from the investigation by iZQC spectroscopy.

In vivo intermolecular zero-quantum coherence MR spectroscopy in the rat spinal cord at 17.6 T: a feasibility study.

OBJECTIVE: The feasibility of in vivo magnetic resonance spectroscopy of the healthy rat spinal cord at 17.6 T using conventional methods and intermolecular zero-quantum coherence (iZQC) spectroscopy is explored and the performance of both approaches is compared. METHODS: Localised spectra were acquired at 17.6 T from three healthy Fisher rats and phantoms with injected iron-oxide particles using the PRESS and a modified HOMOGENIZED sequence. RESULTS: Well-resolved in vivo spectra showing the four singlet resonances of creatine, choline, and N-acetyl aspartate were obtained with both approaches. iZQC spectra were acquired from larger voxels, but did not provide higher sensitivity or resolution in the healthy spinal cord. In the presence of paramagnetic iron-oxide particles, the quality of in vitro spectra acquired with PRESS declined and was strongly dependent on the quality of the local shim. iZQC spectra were not affected by the presence of iron-oxide particles and provided narrow lines (9 Hz) independent of the shim. CONCLUSION: In vivo iZQC spectroscopy of the rat spinal cord is possible. The robustness in presence of local field distortions makes iZQC methods a promising alternative for the investigation of tissue containing labelled cells, implants, or clotted blood. New application of MRS to tissue inaccessible using conventional methods may thus become possible.

Spatially localized intermolecular zero-quantum coherence spectroscopy for in vivo applications.

Magnetic resonance spectroscopy (MRS) techniques that use the distant dipolar field (DDF) to locally refocus inhomogeneous line-broadening promise improved spectral resolution in spatially varying fields. We investigated three possible implementations of localized DDF spectroscopy. Theoretical analysis and phantom experiments at 17.6 T showed that only localization immediately prior to acquisition provides sufficient spatial selectivity and sensitivity for in vivo applications. Spectra from an (8 mm)(3) voxel of the rat brain were acquired in 25 min, and three major metabolites were resolved. In a tumor mouse model, DDF spectra with well-resolved lines can be obtained from significantly larger voxels compared to conventional localized spectroscopy. From an inhomogeneous voxel, improved spectral resolution can be obtained with DDF techniques when a sufficient number of increments are sampled along the second spectral dimension. With fewer increments, measurement time is significantly shortened, and DDF techniques can provide higher signal-to-noise ratio (SNR) efficiency.

Sensitivity to local dipole fields in the CRAZED experiment: an approach to bright spot MRI.

Local dipole fields such as those created by small iron-oxide particles are used to produce regions of low intensity (dark contrast) in many molecular magnetic resonance imaging applications. We have investigated, with computer simulations and experiments at 17.6 T, how the COSY revamped with asymmetric z-gradient echo detection (CRAZED) experiment that selects intermolecular double-quantum coherences can also be used to visualize such local dipole fields. Application of the coherence-selection gradient pulses parallel to the main magnetic field produced similar, dark contrast as conventional gradient echo imaging. Application of the gradient along the magic angle leads to total loss of signal intensity in homogeneous samples. In the presence of local dipole fields, the contrast was inverted and bright signals from the dipoles were observed over a very low background. Both simulations and experiments showed that the signal strongly decreased when a phase-cycle suppressing single-quantum coherences was employed. Therefore, we conclude that most of the signal comes from directly refocused magnetization or intermolecular single-quantum coherences. Finally, we demonstrate that bright contrast from local dipole fields can also be obtained, when the pair of coherence-selection gradient pulses is deliberately mismatched. Both methods allowed visualization of local dipole fields in phantoms in experimental times of about 3 min.