Safety Study of Transcranial Static Magnetic Field Stimulation (tSMS) of the Human Cortex.

BACKGROUND: Transcranial static magnetic field stimulation (tSMS) in humans reduces cortical excitability. OBJECTIVE: The objective of this study was to determine if prolonged tSMS (2 h) could be delivered safely in humans. Safety limits for this technique have not been described. METHODS: tSMS was applied for 2 h with a cylindric magnet on the occiput of 17 healthy subjects. We assessed tSMS-related safety aspects at tissue level by measuring levels of neuron-specific enolase (NSE, a marker of neuronal damage) and S100 (a marker of glial reactivity and damage). We also included an evaluation of cognitive side effects by using a battery of visuomotor and cognitive tests. RESULTS: tSMS did not induce any significant increase in NSE or S100. No cognitive alteration was detected. CONCLUSIONS: Our data indicate that the application of tSMS is safe in healthy human subjects, at least within these parameters.

Adenovirus vector-directed expression of the neurotrophin-3 receptor (TrkC) in mouse astrocytes.

In the present article we report the generation of a neurovirological reagent, an adenovirus vector that efficiently delivers the gene for the Neurotrophin-3 (NT-3) receptor, TrkC. Using this AdTrkC vector, we examined the induction of the expression of the above neurotrophin receptor in pure cultures of mouse astrocytes, a glial cell type that does not constitutively express this gene. Infection of astrocytes at an optimal dose of 100-200 plaque forming units (p.f.u.) per cell, induced expression of specific mRNA, as demonstrated by RT-PCR and Northern blot. This mRNA was translated to produce a mean of 20,157 biologically active receptor molecules per astrocyte with a Kd of 4.1 x 10(-11) M, as demonstrated by 125I-NT-3 binding. After 2D electrophoresis, the mature glycoprotein and some precursors were recognised by antibodies raised against the carboxy-terminal peptide of Trk. Binding of the ligand induced autophosphorylation ofTrkC and 3H-thymidine incorporation in transduced cells. These results demonstrate that our AdTrkC vector efficiently mediates the expression of high-levels of biologically active NT-3 receptors.

Purification and structure of neurostatin, an inhibitor of astrocyte division of mammalian brain.

Neurostatin was originally described as an inhibitor of astroblast and astrocytoma division present in rat brain extracts and immunologically related to the sugar moiety of epidermal growth factor receptor and to blood group antigens. It was purified recently from mammalian brain extracts and characterized as a glycosphingolipid, but its precise structure remained unknown. Neurostatin has now been purified to apparent homogeneity from ganglioside extracts of rat, bovine, and porcine brain. It is cytostatic for astroblasts, C6 glioma cells, and various human astrocytomas grades III and IV, with IC(50) values ranging from 250 to 450 nM, but does not affect the division of primary or transformed fibroblasts up to concentrations >4 microM. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry of purified pig neurostatin showed a molecular ion of 1, 905 Da and ions of 1,863 and 1,934 Da, compatible with a disialoganglioside. Mono- and bidimensional NMR spectra, together with biochemical studies, suggest that neurostatin may be the 9-O-monoacetyl ester of GD1b.

Novel disaccharide inhibitors of human glioma cell division.

Several alpha-L-Fuc-(1-->3)-alpha-D-GlcNAcOC8H17 disaccharide derivatives bearing different hydroxylated alkyl chains, with or without sulfate groups at C-4 and/or C-6 positions of the GlcNAc unit, have been synthesized and tested as inhibitors of human astrocytoma lines U-373 and U-118. The antimitotic activity was dependent on the structure and position of the hydroxylated chain linked to the disaccharide. The compounds with a pentaerythritol or L-glyceryl chain at the C-6 position showed the best inhibitory properties, with an ID50 value of ca. 200 microM. On the contrary, sulfated disaccharide derivatives were inactive. The antimitotic activities of the compounds tested were essentially independent of the mitogen used to stimulate cell division.

Control of glial number: purification from mammalian brain extracts of an inhibitor of astrocyte division.

Inhibitors of astrocyte cell division, immunologically related to the sugar moiety of epidermal growth factor receptor and to blood group antigens, have been purified from mammalian brain extracts. Mass spectra, high resolution proton magnetic resonance spectra, and chemical and enzymic analysis of the purified fraction indicated that the compounds isolated were glycosphingolipids, although signals compatible with the presence of aminoacid residues were also observed. Lectin binding indicated the presence of NAc-Neuraminic acid, NAc-glucosamine, fucose, galactose, and glucose. The inhibitor was cytostatic for astrocytes, C6 glioma cells, and endothelial cells, with approximate ID50 of 250 nM. Primary cultures of fibroblasts or 3T3 cells were not affected up to concentrations of 800 nM. Concentrations of the inhibitor of 800 nM or higher, caused non-specific cytotoxicity. The biological and immunological properties of this brain inhibitor were identical to those of the EGF receptor-related inhibitor previously described with the acronym ERI. Because of its source and cytostatic action, the glial inhibitor has been renamed neurostatin. Rabbit antibodies to neurostatin immunostained astrocytes and neurons, both in culture and in tissue sections.