Biochemically altered myelin triggers autoimmune demyelination.

Although immune attack against central nervous system (CNS) myelin is a central feature of multiple sclerosis (MS), its root cause is unresolved. In this report, we provide direct evidence that subtle biochemical modifications to brain myelin elicit pathological immune responses with radiological and histological properties similar to MS lesions. A subtle myelinopathy induced by abbreviated cuprizone treatment, coupled with subsequent immune stimulation, resulted in lesions of inflammatory demyelination. The degree of myelin injury dictated the resulting immune response; biochemical damage that was too limited or too extensive failed to trigger overt pathology. An inhibitor of peptidyl arginine deiminases (PADs), enzymes that alter myelin structure and correlate with MS lesion severity, mitigated pathology even when administered only during the myelin-altering phase. Moreover, cultured splenocytes were reactive against donor myelin isolates, a response that was substantially muted when splenocytes were exposed to myelin from donors treated with PAD inhibitors. By showing that a primary biochemical myelinopathy can trigger secondary pathological inflammation, "cuprizone autoimmune encephalitis" potentially reconciles conflicting theories about MS pathogenesis and provides a strong rationale for investigating myelin as a primary target for early, preventative therapy.

Seeking the Amygdala: Novel Use of Diffusion Tensor Imaging to Delineate the Basolateral Amygdala.

The amygdaloid complex, including the basolateral nucleus (BLA), contributes crucially to emotional and cognitive brain functions, and is a major target of research in both humans and rodents. However, delineating structural amygdala plasticity in both normal and disease-related contexts using neuroimaging has been hampered by the difficulty of unequivocally identifying the boundaries of the BLA. This challenge is a result of the poor contrast between BLA and the surrounding gray matter, including other amygdala nuclei. Here, we describe a novel diffusion tensor imaging (DTI) approach to enhance contrast, enabling the optimal identification of BLA in the rodent brain from magnetic resonance (MR) images. We employed this methodology together with a slice-shifting approach to accurately measure BLA volumes. We then validated the results by direct comparison to both histological and cellular-identity (parvalbumin)-based conventional techniques for defining BLA in the same brains used for MRI. We also confirmed BLA connectivity targets using DTI-based tractography. The novel approach enables the accurate and reliable delineation of BLA. Because this nucleus is involved in and changed by developmental, degenerative and adaptive processes, the instruments provided here should be highly useful to a broad range of neuroimaging studies. Finally, the principles used here are readily applicable to numerous brain regions and across species.

Monitoring angiogenesis noninvasively with near-infrared spectroscopy.

Near-infrared (NIR) spectroscopy is used to quantify cerebral blood volume (CBV) as a marker of angiogenesis (formation of new blood vessels). Rats are exposed to chronic hypoxia for 3 weeks at half atmospheric pressure to stimulate angiogenesis, and second-differential NIR spectroscopy is used to quantify total cerebral hemoglobin before and after angiogenesis. The cerebral hemoglobin (from broadband NIR spectroscopy), and the large vessel hemoglobin and hematocrit (from blood samples), are used to derive values for the calculation of CBV. The total hemoglobin in brain is 46.6+/-1.9 micromoll (mean+/-SD, n=5) preacclimation and increases by 72% postacclimation. CBV is initially 3.26+/-0.41% v/v and increases by 31% with acclimation. Each individual animal shows a measureable increase in CBV. This study indicates that NIR broadband spectroscopy can be used for repeated measurements of CBV and can be applied as a noninvasive method to study angiogenesis.

Microglia induces Gas1 expression in human brain tumor-initiating cells to reduce tumorigenecity.

We reported previously that microglia decreased the growth of human brain tumor-initiating cells (BTICs). Through microarray analyses of BTICs exposed in vitro to microglia, we found the induction of several genes ascribed to have roles in cell cycle arrest, reduced cell proliferation and differentiation. Herein, we tested the hypothesis that one of these genes, growth arrest specific 1 (Gas1), is a novel growth reduction factor that is induced in BTICs by microglia. We found that microglia increased the expression of Gas1 transcript and protein in glioblastoma patient-derived BTIC lines. Using neurosphere assay we show that RNAi-induced reduction of Gas1 expression in BTICs blunted the microglia-mediated BTIC growth reduction. The role of Gas1 in mediating BTIC growth arrest was further validated using orthotopic brain xenografts in mice. When microglia-induced Gas1-expressing BTIC cells (mGas1-BTICs) were implanted intra-cranially in mice, tumor growth was markedly decreased; this was mirrored in the remarkable increase in survival of mGas1-BT025 and mGas1-BT048 implanted mice, compared to mice implanted with non-microglia-exposed BTIC cells. In conclusion, this study has identified Gas1 as a novel factor and mechanism through which microglia arrest the growth of BTICs for anti-tumor property.

Diffuse encephaloventriculitis and substantial leukoencephalopathy after intraventricular administration of recombinant adenovirus.

OBJECTIVES: The use of recombinant adenovirus as a vehicle for gene transfer into ependymal cells is a potential therapeutic tool for the treatment of various neural disorders. However, gene transfer into the ependymal cells of the ventricular wall is associated with high-level expression of the transferred gene, which declines rapidly. The purpose of this study is to understand the cause of this early decline in gene expression. METHODS: Different doses of adenovirus-expressing beta-galactosidase (Ad-beta-gal) were injected into the lateral brain ventricle of C57BL/6 mice, and the brains were observed histologically and with magnetic resonance (MR) imaging for a month. RESULTS: Inoculation of the lateral ventricle with more than 1 x 10(8) viral particles (2.6 x 10(6) pfu) resulted in a rapid decline of beta -gal expression. MR imaging indicated gradual ventriculomegaly and histological analysis showed the loss of the ependymal cells from the ventricular wall, lymphocytes infiltration near the wall, degeneration of myelinated fibers and apoptosis in the external capsule. Reactive astrocytes proliferated in the external capsule 17 days following inoculation. To avoid this irreversible brain atrophy, the inoculated adenovirus should be reduced to less than 1 x 10(7) particles (2.6 x 10(5) pfu) in mice. DISCUSSION: Our results indicate the presence of a unique and diffuse immune response of the brain; therefore, the clinical use of recombinant virus for intraventricular gene transfer must be carefully evaluated.

Diffusion-weighted imaging in the follow-up of treated high-grade gliomas: tumor recurrence versus radiation injury.

BACKGROUND AND PURPOSE: Diffusion-weighted (DW) MR imaging is a means to characterize and differentiate morphologic features, including edema, necrosis, and tumor tissue, by measuring differences in apparent diffusion coefficient (ADC). We hypothesized that DW imaging has the potential to differentiate recurrent or progressive tumor growth from treatment-induced damage to brain parenchyma in high-grade gliomas after radiation therapy. METHODS: We retrospectively reviewed follow-up conventional and DW MR images obtained starting 1 month after completion of radiation treatment with or without chemotherapy for histologically proved high-grade gliomas. Eighteen patients with areas of abnormal enhancing tissue were identified. ADC maps were calculated from echo-planar DW images, and mean ADC values and ADC ratios (ADC of enhancing lesion to ADC of contralateral white matter) were compared with final diagnosis. Recurrence was established by histologic examination or by clinical course and a combination of imaging studies. RESULTS: Recurrence and nonrecurrence could be differentiated by using mean ADC values and ADC ratios. ADC ratios in the recurrence group showed significantly lower values (mean +/- SD, 1.43 +/- 0.11) than those of the nonrecurrence group (1.82 +/- 0.07, P <.001). Mean ADCs of the recurrent tumors (mean +/- SD, 1.18 +/- 0.13 x 10(-3) mm/s(2)) were significantly lower than those of the nonrecurrence group (1.40 +/- 0.17 x 10(-3) mm/s(2), P <.006). CONCLUSION: Assessment of ADC ratios of enhancing regions in the follow-up of treated high-grade gliomas is useful in differentiating radiation effects from tumor recurrence or progression.