NMR-based metabolomics in pediatric drug resistant epilepsy - preliminary results.

Epilepsy in children is the most frequent, heterogeneous and difficult to classify chronic neurologic condition with the etiology found in 35-40% of patients. Our aim is to detect the metabolic differences between the epileptic children and the children with no neurological abnormalities in order to define the metabolic background for therapy monitoring. The studied group included 28 epilepsy patients (median age 12 months) examined with a diagnostic protocol including EEG, videoEEG, 24-hour-EEG, tests for inborn errors of metabolism, chromosomal analysis and molecular study. The reference group consisted of 20 patients (median age 20 months) with no neurological symptoms, no development delay nor chronic diseases. 1H-NMR serum spectra were acquired on 400 MHz spectrometer and analyzed using multivariate and univariate approach with the application of correction for age variation. The epilepsy group was characterized by increased levels of serum N-acetyl-glycoproteins, lactate, creatine, glycine and lipids, whereas the levels of citrate were decreased as compared to the reference group. Choline, lactate, formate and dimethylsulfone were significantly correlated with age. NMR-based metabolomics could provide information on the dynamic metabolic processes in drug-resistant epilepsy yielding not only disease-specific biomarkers but also profound insights into the disease course, treatment effects or drug toxicity.

A three-dimensional stereotaxic atlas of the gray short-tailed opossum (Monodelphis domestica) brain.

The gray short-tailed opossum (Monodelphis domestica) is a small marsupial gaining recognition as a laboratory animal in biomedical research. Despite numerous studies on opossum neuroanatomy, a consistent and comprehensive neuroanatomical reference for this species is still missing. Here we present the first three-dimensional, multimodal atlas of the Monodelphis opossum brain. It is based on four complementary imaging modalities: high resolution ex vivo magnetic resonance images, micro-computed tomography scans of the cranium, images of the face of the cutting block, and series of sections stained with the Nissl method and for myelinated fibers. Individual imaging modalities were reconstructed into a three-dimensional form and then registered to the MR image by means of affine and deformable registration routines. Based on a superimposition of the 3D images, 113 anatomical structures were demarcated and the volumes of individual regions were measured. The stereotaxic coordinate system was defined using a set of cranial landmarks: interaural line, bregma, and lambda, which allows for easy expression of any location within the brain with respect to the skull. The atlas is released under the Creative Commons license and available through various digital atlasing web services.

Combined orcein and martius scarlet blue (OMSB) staining for qualitative and quantitative analyses of atherosclerotic plaques in brachiocephalic arteries in apoE/LDLR-/- mice.

Numerous cellular and extracellular components should be analyzed in sections of atherosclerotic plaques to assess atherosclerosis progression and vulnerability. Here, we combined orcein (O) staining for elastic fibers and martius scarlet blue (MSB) polychrome to visualize various morphological contents of plaque in brachiocephalic arteries (BCA) of apoE/LDLR-/- mice. Elastic fibers (including broken elastic laminae and 'buried' fibrous caps) were stained purple and they could be easily distinguished from collagen fibers (blue). Orcein allowed clear identification of even the finest elastic fibers. Erythrocytes were stained yellow and they could easily be discerned from mature fibrin (red). Old fibrin tends to acquire blue color. The method of OMSB staining is simple, takes less than 1 h to perform and can be adapted to automatic stainers. Most importantly, the color separation is good enough to allow digital automatic segmentation of specific components in tissue section and quantitative analysis of the plaque constituents. OMSB was used to compare atherosclerotic plaques in proximal and distal regions of BCA in apoE/LDLR-/- mice. In conclusion, OMSB staining represents a novel staining that could be routinely used for qualitative and quantitative microscopic assessments of formaldehyde-fixed and paraffin-embedded sections of arteries with atherosclerotic lesions.

Four-and-one-half years' experience in monitoring of reproducibility of an MR spectroscopy system--application of in vitro results to interpretation of in vivo data.

The primary purpose of this work was to assess long-term in vitro reproducibility of metabolite levels measured using 1H MRS (proton magnetic resonance spectroscopy). The secondary purpose was to use the in vitro results for interpretation of 1H MRS in vivo spectra acquired from patients diagnosed with Canavan disease. 1H MRS measurements were performed in the period from April 2006 to September 2010. 118 short and 116 long echo spectra were acquired from a stable phantom during this period. Change-point analysis of the in vitro N-acetylaspartate levels was exploited in the computation of fT factor (ratio of the actual to the reference N-acetylaspartate level normalized by the reciprocity principle). This coefficient was utilized in the interpretation of in vivo spectra analyzed using absolute reference technique. The monitored time period was divided into six time intervals based on short echo in vitro data (seven time intervals based on long echo in vitro data) characterized by fT coefficient ranging from 0.97 to 1.09 (based on short echo data) and from 1.0 to 1.11 (based on long echo data). Application of this coefficient to interpretation of in vivo spectra confirmed increased N-acetylaspartate level in Canavan disease. Long-term monitoring of an MRS system reproducibility, allowing for absolute referencing of metabolite levels, facilitates interpretation of metabolic changes in white matter disorders.

Magnetic resonance spectroscopy as a predictor of conversion of mild cognitive impairment to dementia.

BACKGROUND AND PURPOSE: Proton magnetic resonance spectroscopy ((1)H-MRS) is one of the imaging techniques that could be potentially useful for identification of patients with mild cognitive impairment (MCI) who are at increased risk of developing dementia. The aim of the study was to investigate if there is a difference in brain metabolism between stable MCI patients and converters to dementia and if a use of (1)H-MRS can predict the conversion from MCI to dementia. MATERIAL AND METHODS: Forty-one amnestic MCI patients and 35 cognitively unimpaired controls were examined by (1)H-MRS (TE - 35 ms) at baseline. Metabolite ratios (NAA/Cr, Cho/Cr, mI/Cr, Glx/Cr, NAA/Cho) were estimated in four different brain regions: posterior cingulate gyrus (PCG), left hippocampus (LH), cortical area of right (RPL) and left parietal lobe (LPL). Participants were followed up for a period of 12 months. RESULTS: Twelve subjects with MCI progressed to Alzheimer's disease (AD) after one year. Analysis showed that the NAA/Cr ratio in the LH was significantly lower in MCI patients than in controls (p=0.008), but there were no differences in metabolite ratios at baseline between MCI converters and stable subjects. mI/Cr ratio in RPL predicted the conversion to AD with sensitivity 70% and specificity 85% (p<0.0004). Coexistence of diabetes improved prediction, yielding 70% sensitivity and 96% specificity (p<0.0001). CONCLUSIONS: (1)H-MRS in MCI can be a predictor of cognitive decline and conversion to dementia, especially in MCI patients with coexisting diabetes. Further studies are needed to confirm this finding.

Application of magnetic resonance diffusion anisotropy imaging for the assessment neuroprotecting effects of MPEP, a selective mGluR5 antagonist, on the rat spinal cord injury in vivo.

Magnetic resonance diffusion anisotropy imaging (DAI) of the rat spinal cord after contusion using weight-drop method was used to study the neuroprotecting effect of 2-methyl-6-(phenylethynyl)-pyridine (MPEP), an mGluR5 receptor antagonist. Eighteen rats were used, divided into 3 groups of 6 animals: a reference group without any operation, a control group with injury and a test group with injury and MPEP. DAI was performed at 4.7 T at 1 h, 24 h, 48 h and 7 day after the injury. Locomotor function was evaluated using Basso, Beattie and Bresnahan (BBB) open field locomotor activity test each day starting one day after the injury. DAI results confirm positive effect of MPEP on the limitation of secondary excitotoxic injury in the spinal cord.