Impaired eyeblink conditioning and decreased hippocampal volume in PDAPP V717F mice.

We examined heterozygous transgenic (Tg) mice that overexpress V717F amyloid precursor protein (APP) for delay eyeblink conditioning (EBC) and hippocampal volume with magnetic resonance imaging (MRI). Platelet-derived APP mice were significantly impaired on EBC relative to wild type (WT) litter-mate controls. T2-weighted spin echo images (62.5 x 125 x 500 microm) of the same mice were acquired under anesthesia using a 9.4T magnet. Tg mice had hippocampal to brain volume ratios that were significantly smaller than WT controls (31% smaller in the rostral dorsal hippocampus, 13-22% smaller among equal dorsal-ventral thirds of a caudal section). These results indicate that overexpression of APP or beta amyloid profoundly affects learning and memory and hippocampal volume. The results also indicate that eyeblink conditioning and quantitative MRI in mice may be useful assays to follow the progression of disease-related changes, and to test the effectiveness of potential therapeutics against Alzheimer's disease.

Monitoring of brain water by chemical shift imaging during ammonia-induced brain swelling in rats after portacaval anastomosis.

Brain edema is a leading cause of death in acute liver failure (ALF). In experimental models of ALF, an increase in the content of brain water has been inferred indirectly by measuring intracranial pressure or determined directly via analysis of brain tissue postmortem. In this study, noninvasive proton two-dimensional chemical shift imaging (2-D CSI) was used to follow the time course of the development of brain edema in a well characterized model, namely ammonium acetate infusion into rats 48 to 72 h after portacaval anastomosis (PCA). Clear differences between control and experimental rat brains were observed, with an increase of brain water signal only in the parietal cortex of the PCA + ammonia group. Selective swelling of the cerebral cortex points to a cytotoxic mechanism in the evolution of brain edema in this model. CSI signal enhancement was much greater than the gravimetrically determined water content increase. The significantly greater signal change observed with 2-D CSI may reflect enhanced proton density that results from increased water content as well as edema-related alterations in water relaxation times.

Optimized distortion correction technique for echo planar imaging.

A new phase-shifted EPI pulse sequence is described that encodes EPI phase errors due to all off-resonance factors, including B(o) field inhomogeneity, eddy current effects, and gradient waveform imperfections. Combined with the previously proposed multichannel modulation postprocessing algorithm (Chen and Wyrwicz, MRM 1999;41:1206-1213), the encoded phase error information can be used to effectively remove geometric distortions in subsequent EPI scans. The proposed EPI distortion correction technique has been shown to be effective in removing distortions due to gradient waveform imperfections and phase gradient-induced eddy current effects. In addition, this new method retains advantages of the earlier method, such as simultaneous correction of different off-resonance factors without use of a complicated phase unwrapping procedure. The effectiveness of this technique is illustrated with EPI studies on phantoms and animal subjects. Implementation to different versions of EPI sequences is also described. Magn Reson Med 45:525-528, 2001.

fMRI of visual system activation in the conscious rabbit.

A conscious rabbit preparation developed for fMRI, and the results from visual stimulation studies at a 4.7T magnetic field are described. The rabbit is ideal for these experiments because of its natural tolerance for restraint. High spatial and temporal resolution magnetic resonance images, without movement artifacts, were obtained during long periods of restraint. Functional activation in primary visual cortex and lateral geniculate nucleus (LGN) were reproducibly observed in response to light stimulus. In comparison to existing anesthetized animal models, a functional response free of the anesthetic modulation can be recorded with the new approach. The conscious animal model can be applied to functional studies of sensory systems, learning and memory, and drug-induced cerebral activation.

In vitro 1H and 31P NMR spectroscopic evidence of multiple aberrant biochemical pathways in murine trisomy 16 brain development.

Nuclear magnetic resonance (NMR) spectroscopy was used to evaluate cytosolic compounds and membrane phospholipids simultaneously in trisomy 16 (Ts16) and euploid (control) murine brain at fetal day 15 in order to examine the cellular biochemistry that underlies the neurodevelopmental consequences of chromosome triplication in this model of Down syndrome (DS). Proton NMR spectroscopic analysis of brain tissue extracts demonstrated decreased levels of choline and increased levels of myo-inositol (MI) in Ts16 brains compared with control. These data are consistent with the cholinergic deficits and elevated MI levels previously described in Ts16. Compared with euploid brains. Ts16 brains also possess higher levels of creatine, adenosine, and tyrosine. Increased levels of MI and creatine, compounds that are localized to glia, imply abnormalities in the trophic environment of Ts16 brain. Phosphorus NMR spectroscopic analysis of extracts further revealed elevated levels of anionic phospholipid membrane components, such as phosphatidylinositol (PtdIno) and phosphatidylethanolamine, in Ts16 brains. Since these compounds are confined to the inner leaflet of the membrane, the findings suggest that membrane composition is altered specifically in the cytosolic bilayer at this stage. Together our proton and phosphorus NMR spectroscopic results indicate that multiple biochemical pathways are affected in Ts16 brain development. Understanding the effects of these aberrations may elucidate the processes that lead to neural dysfunction and Alzheimer's disease (AD) neuropathology in DS individuals.

Quantitative 19F NMR study of trifluorothymidine metabolism in rat brain.

Metabolism of trifluorothymidine (TFT) and its transport across the blood-brain barrier (BBB) has been measured quantitatively in rats by fluorine-19 nuclear magnetic resonance spectroscopy ((19)F NMR). It is demonstrated that TFT crosses the BBB in micromolar quantities and is metabolized in brain tissue primarily to its free base trifluoromethyluracil (TFMU) by the enzyme thymidine phosphorylase (TP). It is further proposed that the rate of TFMU production can be used as a measure of cerebral TP. The glycols of both TFMU, and to a lesser degree TFT, are generated via an oxidative route. In contrast, the major pathway for hepatic metabolism of this compound is through reduction of the nitrogen base moiety and generation of 5-6-dihydro species followed by ring degradation. Thus, in addition to TFMU as well as the dihydroxy (glycol)-, and the dihydro-species of both TFT and TFMU, alpha-trifluoromethyl-beta-ureidopropionic acid (F(3)MUPA) and alpha-trifluoromethyl-beta-alanine (F(3)MBA) were detected in liver extracts. The total metabolite levels in liver were 2-5 times higher than in the brain. Low levels of fluoride ion were detected in all the extracts from brain and liver, as well as blood and urine. This study characterizes TFT as a potential chemotherapeutic agent for use against brain tumors.

Removal of intravoxel dephasing artifact in gradient-echo images using a field-map based RF refocusing technique.

A technique is proposed to compensate for the slice dephasing artifact and improve the signal-to-noise ratio (SNR) of gradient-echo images. This method is composed of two components: mapping of the internal gradient and design of the slice-selective radiofrequency (RF) pulse. The RF pulse is designed with its phase response as the negative of the product of a chosen echo time and the intravoxel internal gradient profile in a specified region of interest (ROI). The designed RF pulse can refocus the spin phases at a selected echo time and therefore effectively recover the signal loss due to both linear and nonlinear internal gradients. Principles, implementation, and application of the method are described in this note. Magn Reson Med 42:807-812, 1999.

Correction for EPI distortions using multi-echo gradient-echo imaging.

A novel and effective technique is described for distortion correction in echo planar imaging (EPI) utilizing the field maps derived from multi-echo gradient-echo images. The distortions from different off-resonance related factors such as field inhomogeneity, eddy current effect, radiofrequency pulse frequency offset, and chemical shift effect can be simultaneously reduced to a great extent. With the proposed post-processing algorithm of multi-channel modulation, distortions may be corrected without unwrapping the phase discontinuities in the derived field map, a process that usually restricts the application of other field map-based correction methods. Results from phantom and animal experiments at 4.7 T demonstrate the efficiency of the method in reducing the geometrical distortions in gradient-echo EPI.

In vitro proton and phosphorus NMR spectroscopic analysis of murine (C57Bl/6J) brain development.

We report for the first time in vitro proton and phosphorus NMR spectroscopic analyses of murine brain development from fetal to adult stages. Chloroform-methanol extracts from C57B16/J mouse brain, at ages ranging from 15 days in utero (F15) to adult, permitted the simultaneous investigation of both cytosolic and membrane phospholipid compartments. The protein content of murine brain was determined and used for quantitation of individual metabolite levels. Proton NMR spectroscopy revealed that NAA, considered a neuronal marker, is undetectable at F15. Glutamate, GABA and creatine, however, are present at this time. All four compounds reach maximum levels at 21 days postnatal (P21). Choline and alanine levels are at their peak in fetal brain and progressively fall as the brain develops. Phosphorus NMR spectroscopy shows that phosphatidylcholine, phosphatidylinositol, sphingomyelin, and phosphatidylserine increase steadily from F15 to P21.

Characterization of the cerebral distribution of general anesthetics in vivo by two-dimensional 19F chemical shift imaging.

The distribution of two fluorinated anesthetics, halothane and isoflurane, in rabbit brain was mapped during the course of their uptake using 19F NMR chemical shift imaging techniques. Use of a short phase-encoding time and free induction decay acquisition minimized T2-related signal losses in two-dimensional chemical shift imaging. Localization of the brain and exclusion of surrounding tissues was achieved by the use of a surface coil. The spatial distribution of halothane and isoflurane in the brain was nonuniform and both anesthetics were detected predominantly in the cerebral cortex. During the early stages of uptake, anesthetic concentration in the brain was extremely low, but it continued to increase for several hours with sustained anesthetic delivery. Time-dependent variations in anesthetic concentration were observed in different brain regions.

In vivo 19F-NMR spectroscopic study of halothane uptake in rabbit brain.

Uptake of a fluorinated anesthetic, halothane, in rabbit brain and blood was studied using 19F-NMR spectroscopic techniques. Localized one-dimensional chemical shift imaging and non-localized one-pulse sequence were used to measure brain uptake kinetics in vivo. Halothane signal was found predominantly in the cerebral cortex. Uptake in the brain followed a first-order biexponential kinetics. The average half-lives were 4 min and 70 min, respectively, for the 'fast' and 'slow' phases of the uptake. Uptake in the arterial blood was also biexponential. However, equilibration of halothane in the brain considerably lagged behind that in arterial blood. This delay was ascribed to a 'restricted diffusion' of the anesthetic molecule into brain tissue.

In vivo 19F one-dimensional chemical shift imaging study of isoflurane uptake in rabbit brain.

19F one-dimensional chemical shift imaging and one-pulse techniques were used to follow the uptake of isoflurane in rabbit brain in vivo. Brain isoflurane appeared mainly in two adjacent slices of cerebral cortex. Both slices showed increase in anesthetic content at similar rates. A biphasic first-order uptake pattern with an initial fast component and a slower rate for the duration of the uptake was observed by the one-pulse technique. Only the slower kinetic was followed by chemical shift imaging due to constraints on temporal resolution. Anesthetic levels in arterial blood reached equilibrium significantly earlier than in brain, suggesting a 'restricted' diffusion into brain tissue for this agent.

Cerebral metabolism of fluorodeoxyglucose measured with 19F NMR spectroscopy.

Brain extracts from rats given various doses of 2-fluoro-2-deoxy-D-glucose ranging from 20 to 300 mg/kg were examined with 19F NMR spectroscopy for fluorinated metabolic products and compared with the in vivo 19F spectra. Based on the analysis of these spectra and comparison with synthetic reference compounds, 2-fluoro-2-deoxy-D-glucose 6-phosphate and 2-fluoro-2-deoxy-D-mannose 6-phosphate were identified as the major metabolites present. 2-Fluoro-2-deoxy-D-6-phosphogluconate was detected only at high doses of 2-fluoro-2-deoxy-D-glucose administered, along with non-phosphorylated 2-fluoro-2-deoxy-D-glucose and 2-fluoro-2-deoxy-D-mannose. Small amounts of two other compounds, which were not identified, were also observed. Presence of the 2-fluoro-2-deoxy-D-mannose 6-phosphate is the result of 2-fluoro-2-deoxy-D-glucose 6-phosphate isomerization at the C2 position catalyzed by phosphoglucose isomerase. Results from the in vitro enzymatic reaction provide support for this hypothesis. Presence of 2-fluoro-2-deoxy-D-mannose may be the result of 2-fluoro-2-deoxy-D-glucose isomerization at the C2 position catalyzed by glucose isomerase or the result of the dephosphorylation of the 2-fluoro-2-deoxy-D-glucose 6-phosphate.

Proton nuclear magnetic resonance spectroscopy of plasma lipoproteins in head and neck cancer patients.

Proton nuclear magnetic resonance spectroscopy was used to evaluate changes in plasma lipoproteins in patients with squamous cell head and neck cancer. Plasma from 14 patients was analyzed by proton nuclear magnetic resonance spectroscopy, and line widths and fast and slow methyl and methylene transverse relaxation values were obtained. In addition, the lipid and lipoprotein concentrations in the sera of these patients were measured by standard biochemical techniques. Preliminary results suggested that squamous cell carcinomas of the head and neck with nodal metastases are associated with measureable changes in slow methylene transverse relaxation values, as compared with controls. These findings indicate the presence of a new lipoprotein complex in patients with squamous cell carcinomas of the head and neck with nodal metastases.

Determination of halothane distribution in the rat head using 19F NMR technique.

The distribution of halothane in the rat head was examined with 19F NMR rotating-frame zuegmatography and 2DFT 19F NMR imaging. The rotating frame experiments were conducted at varying times following anesthesia to assess the time dependence of the halothane distribution. The results of these experiments demonstrate that halothane and halothane metabolite are unequally partitioned through the rat tissues examined. 19F spin-echo imaging experiments were conducted immediately following anesthesia. The results of these experiments are compared with those of the spectroscopic technique.

In vivo 19F-NMR study of halothane distribution in brain.

Halothane distribution and elimination from rabbit brain was studied in vivo using 19F-NMR spectroscopy. Two exponential decay functions for the anesthetic were observed in the clearance curve. They are assigned to halothane in brain held in two distinct chemical environments characterized by different chemical shifts, and half-lives (25 and 320 min). A nonvolatile halothane metabolite with a half-life of several days was found to be present in rabbit brains. The in vivo results were corroborated by ex vivo experiments on excised brain tissue. Halothane was distributed in all of the major cell subfractions, whereas the metabolite was present predominantly in the cytoplasm.

In vivo 19F-NMR study of isoflurane elimination from brain.

The time course of isoflurane elimination from rabbit brain was studied in vivo with 19F-NMR spectroscopy. Two exponential decay functions with different time constants were observed and assigned to two distinct brain compartments. Isoflurane has a 26 min time constant for one compartment (similar to a value of 25 min with halothane) but 174 min in the second one, compared with 320 min for halothane. The shorter half-life for isoflurane may be due to lower solubility of this agent in brain tissue. Comparison of isoflurane 19F chemical shifts in solvents in isolated brain lipids and in whole brain tissue indicates that the anesthetic present in the brain exists in a single environment (on the NMR time scale), which is a weighted average of both hydrophilic and hydrophobic environments.

Noninvasive observations of fluorinated anesthetics in rabbit brain by fluorine-19 nuclear magnetic resonance.

Fluorinated anesthetics were observed noninvasively in the brain of intact rabbits with fluorine-19 nuclear magnetic resonance spectroscopy. High-resolution fluorine-19 spectra of halothane, methoxyflurane, and isoflurane were obtained with a surface coil centered over the calvarium. Elimination of halothane from the brain was also monitored by this technique. Residual fluorine-19 signals from halothane (or a metabolite) could be detected as long as 98 hours after termination of anesthesia. These observations demonstrate the feasibility of using this technique to study the fate of fluorinated anesthetics in live mammals.