In vivo quantitative 1H MRS of cerebellum and evaluation of quantitation reproducibility by simulation of different levels of noise and spectral resolution

A quantitative analysis of cerebellar metabolites in normal subjects has been performed by proton MR spectroscopy (MRS) with relaxation time correction. Quantitation was carried out in seven healthy human subjects with the well-established LCModel program. The prior knowledge utilized for quantitation was obtained from solutions containing the major brain metabolites and MRS investigated under the same experimental conditions. The tissue water signal was used as an internal standard for the in vivo studies. Both in vitro (for the prior knowledge template) and in vivo data were acquired separately at 1.5 T by PRESS sequence (TR, 1500 ms; TE, 30 ms). The absolute concentration of main cerebellar metabolites was corrected for relaxation time effects. Different noise and line broadening conditions were considered and simulated in the spectral processing in order to evaluate the effect of spectral quality on the concentration estimates.     

An automatic cerebellum extraction method in T1-weighted brain MR images using an active contour model with a shape prior

Purpose

The objective of this paper was to automatically segment the cerebellum from T1-weighted human brain magnetic resonance (MR) images.

Materials and Methods

The proposed method constructs a cerebellum template using five sets of 3-T MR imaging (MRI) data, which are used to determine the initial position and the shape prior of the cerebellum for the active contour model. Our formulation includes the active contour model with shape prior, which thereby maintains the shape of the template. The proposed active contour model is sequentially applied to sagittal-, coronal- and transverse-view images. To evaluate the proposed method, it is applied to BrainWeb data and a 3-T MRI data set and compared with FreeSurfer with respect to performance assessment metrics.

Results

The segmented cerebellum was compared with the results from FreeSurfer. Using the manually segmented cerebellum as reference, we measured the average Jaccard coefficients of the proposed method, which were 0.882 and 0.885 for the BrainWeb data and 3-T MRI data set, respectively.

Conclusion

We presented the active contour model with shape prior for extracting the cerebellum from T1-weighted brain MR images. The proposed method yielded a robust and accurate segmentation result.     

Magnetization transfer MRI of mouse brain reveals areas of high neural density

Extending applications of magnetization transfer contrast (MTC) in magnetic resonance imaging (MRI) of the human central nervous system, this work quantitatively describes MTC of the murine brain. As a novel finding, complementing T₁- and T₂-weighted MRI, MTC allows for the distinction of densely packed gray matter from normal gray and white matter. Examples include the Purkinje cell layer and the granular cell layer in the mouse cerebellum as well as the delineation of the CA3 subfield of the hippocampus relative to surrounding hippocampal gray matter and white matter tracts such as the hippocampal fimbria. Using a kainate lesion model, the CA3 hyperintensities in MTC and T₁-weighted MRI are assigned to the densely packed somata of pyramidal cells.     

Bootstrap classification and point-based feature selection from age-staged mouse cerebellum tissues of matrix assisted laser desorption/ionization mass spectra using a fuzzy rule-building expert system

A bootstrap method for point-based detection of candidate biomarker peaks has been developed from pattern classifiers. Point-based detection methods are advantageous in comparison to peak-based methods. Peak determination and selection are problematic when spectral peaks are not baseline resolved or on a varying baseline. The benefit of point-based detection is that peaks can be globally determined from the characteristic features of the entire data set (i.e., subsets of candidate points) as opposed to the traditional method of selecting peaks from individual spectra and then combining the peak list into a data set. The point-based method is demonstrated to be more effective and efficient using a synthetic data set when compared to using Mahalanobis distance for feature selection. In addition, probabilities that characterize the uniqueness of the peaks are determined.This method was applied for detecting peaks that characterize age-specific patterns of protein expression of developing and adult mouse cerebella from matrix assisted laser desorption/ionization (MALDI) mass spectrometry (MS) data. The mice comprised three age groups: 42 adults, 19 14-day-old pups, and 16 7-day-old pups. Three sequential spectra were obtained from each tissue section to yield 126, 57 and 48 spectra for adult, 14-day-old pup, and 7-day-old pup spectra, respectively. Each spectrum comprised 71,879 mass measurements in a range of 3.5-50 kDa. A previous study revealed that 846 unique peaks were detected that were consistent for 50% of the mice in each age group (C. Laurent, D.F. Levinson, S.A. Schwartz, P.B. Harrington, S.P. Markey, R.M. Caprioli, P. Levitt, Direct profiling of the cerebellum by MALDI MS: a methodological study in postnatal and adult mouse, J. Neurosci. Res. 81 (2005) 613-621.).A fuzzy rule-building expert system (FuRES) was applied to investigate the correlation of age with features in the MS data. FuRES detected two outlier pup-14 spectra. Prediction was evaluated using 100 bootstrap samples of 2 Latin-partitions (i.e., 50:50 split between training and prediction set) of the mice. The spectra without the outliers yielded classification rates of 99.1 ± 0.1%, 90.1 ± 0.8%, and 97.0 ± 0.6% for adults, 14-day-old pups, and 7-day-old pups, respectively. At a 95% level of significance, 100 bootstrap samples disclosed 35 adult and 21 pup distinguishing peaks for separating adults from pups; and 8 14-day-old and 15 7-day-old predictive peaks for separating 14-day-old pup from 7-day-old pup spectra. A compressed matrix comprising 40,393 points that were outside the 95% confidence intervals of one of the two FuRES discriminants was evaluated and the classification improved significantly for all classes. When peaks that satisfied a quality criterion were integrated, the 55 integrated peak areas furnished significantly improved classification for all classes: the selected peak areas furnished classification rates of 100%, 97.3 ± 0.6%, and 97.4 ± 0.3% for adult, 14-day-old pups, and 7-day-old pups using 100 bootstrap Latin partitions evaluations with the predictions averaged. When the bootstrap size was increased to 1000 samples, the results were not significantly affected. The FuRES predictions were consistent with those obtained by discriminant partial least squares (DPLS) classifications.     

Application of Chip-Based Nanoelectrospray Ion Trap Mass Spectrometry to Compositional and Structural Analysis of Gangliosides in Human Fetal Cerebellum

Two native ganglioside mixtures from normal human fetal cerebellum in the 15th (Cc15) and 40th (Cc40) gestational week were subjected to NanoMate high capacity ion trap (HCT) mass spectrometric (MS) and collision induced dissociation (CID) tandem MS (MS²) analysis under thoroughly optimized experimental conditionns. An total of 56 different species were identified in Cc15 and 54 in Cc40. By employing CID MS² molecular ions, related GD1 (d18:1/20:0) and GM2 (d18:1/19:0) species were structurally characterized in a high throughput mode. The method provided elevated ionization efficiency, high speed of analysis, almost 100% reproducibility at sample consumption per experiment situated in the femtomole range.     

In Vivo Determination of Reduced Thiols in Rat Cerebellum Paraflocculus Following Salicylate-Induced Tinnitus by Fluorescence

The increasing number of patients suffering from tinnitus may be explained by an imbalance in the antioxidant defense system and reactive oxygen species formation, suggesting the importance of the redox homeostasis in this condition. Endogenous biothiols play important roles in maintaining redox homeostasis. Hence, to understand the role of biothiols in the pathological process of tinnitus, this study demonstrates an in vivo method for monitoring the concentrations of biothiols in the paraflocculus of the rat cerebellum during tinnitus induced by the injection of salicylate. Resorufin-based fluorescent probe 1 was used as the selective probe with in vivo microdialysis. Probe 1 was premixed with microdialysates from the paraflocculus of the rat cerebellum and transferred into a cuvette for continuous-flow fluorescence. A linear relationship between the fluorescence and the concentrations of cysteine, homocysteine, and glutathione was from 1 to 15?µM, with detection limits of 128, 148, and 183?nM, respectively. The basal level of total reduced biothiols in the paraflocculus of the rat cerebellum microdialysate was determined to be 10.62?±?1.34?µM based on the calibration curve for homocysteine. The injection of sodium salicylate into the animals significantly decreased the reduced biothiol concentrations in the paraflocculus of the rat cerebellum from 60?min, reaching 39.5?±?6.20% of the basal level. In contrast, at 120, 180, and 300?min, the concentrations of reduced biothiols were 55.73?±?9.35, 58.26?±?9.56, and 91.69?±?6.91% of the basal level, respectively. These results imply that the decrease in reduced biothiols in the paraflocculus of the rat cerebellum may be associated with salicylate-induced tinnitus. This study offers a new platform for in vivo monitoring of reduced biothiols in the paraflocculus of the rat cerebellum following salicylate-induced tinnitus and may be useful for the investigation of this condition.     

Determination of Metabolites in the Cerebellum of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin Exposed Mice by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin elicits many harmful effects in tissues. Metabolomic changes and the associated pathway alterations caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin in the cerebellum, an area thought to be less affected by environmental alterations, remain unknown. Here, metabolomics was performed to identify endogenous metabolites that were associated with 2,3,7,8-tetrachlorodibenzo-p-dioxin in the cerebellum of 2,3,7,8-tetrachlorodibenzo-p-dioxin-treated mice using Fourier transform ion cyclotron resonance mass spectrometry. Distinct peaks were located in two mass ranges, 210 m/z–420 m/z and 450 m/z–570 m/z. In principal component space, the high-dose group was clearly separated from the control group. Six metabolites associated with 2,3,7,8-tetrachlorodibenzo-p-dioxin dose were identified. The metabolite 1-palmitoyl lysophosphatidic acid increased with increasing doses of 2,3,7,8-tetrachlorodibenzo-p-dioxin, indicating activation of the rat sarcoma pathway. Biosynthesis of the unsaturated fatty acid 18-hydroxyoleate was inhibited upon 2,3,7,8-tetrachlorodibenzo-p-dioxin exposure. The decrease in N-arachidonoyl taurine, implies that taurine increased, suggesting inhibition of neuronal signal transmission. A decrease in N-acetyl-aspartyl-glutamate has been associated with injury of the cerebellum through activation of N-methyl-D-aspartic acid receptors. An increase in glycerophosphoinositol suggests damage to blood–brain barrier function, and changes in purine metabolism were observed because inosine increased following 2,3,7,8-tetrachlorodibenzo-p-dioxin exposure. These results suggest that 2,3,7,8-tetrachlorodibenzo-p-dioxin activates the rat sarcoma pathway, alters fatty acid biosynthesis and purine metabolism, inhibits neurotransmitter systems, and is harmful to blood–brain barrier function in the cerebellum.     

ICP-AES法测定胎儿小脑组织中生物必需常量元素K、Na、Mg、P的研究

研究了用微波消解处理样品，ICP－AES法直接测定胎儿小脑组织中生物必需常量元素K、Na、Mg、P的新方法，该方法简便，快速，准确，准确，对实际样品测定，结果令人满意。