Subcellular Localization of Matrin 3 Containing Mutations Associated with ALS and Distal Myopathy.

BACKGROUND: Mutations in Matrin 3 [MATR3], an RNA- and DNA-binding protein normally localized to the nucleus, have been linked to amyotrophic lateral sclerosis (ALS) and distal myopathies. In the present study, we have used transient transfection of cultured cell lines to examine the impact of different disease-causing mutations on the localization of Matrin 3 within cells. RESULTS: Using CHO and human H4 neuroglioma cell models, we find that ALS/myopathy mutations do not produce profound changes in the localization of the protein. Although we did observe variable levels of Matrin 3 in the cytoplasm either by immunostaining or visualization of fluorescently-tagged protein, the majority of cells expressing either wild-type (WT) or mutant Matrin 3 showed nuclear localization of the protein. When cytoplasmic immunostaining, or fusion protein fluorescence, was seen in the cytoplasm, the stronger intensity of staining or fluorescence was usually evident in the nucleus. In ~80% of cells treated with sodium arsenite (Ars) to induce cytoplasmic stress granules, the nuclear localization of WT and F115C mutant Matrin 3 was not disturbed. Notably, over-expression of mutant Matrin 3 did not induce the formation of obvious large inclusion-like structures in either the cytoplasm or nucleus. CONCLUSIONS: Our findings indicate that mutations in Matrin 3 that are associated with ALS and myopathy do not dramatically alter the normal localization of the protein or readily induce inclusion formation.

Toughening of dental glass ionomer cements with reactive glass fibres.

A fibre reinforced glass ionomer cement (FRGIC) for dental applications was loaded with 20 vol% short fibres (430 microm) with a glass composition in the system SiO(2)-Al(2)O(3)-CaF(2)-Na(3)AlF(6). The fracture toughness and the total energy release rate were examined. A 20% anisotropic fibre alignment was observed, perpendicular to the loading direction. An increase of fracture toughness of 140% and of total energy release rate of 440% was achieved compared to the unreinforced glass ionomer cement. Matrix-fibre interface reaction is supposed to exert the major influence on mechanical behaviour of FRGIC by controlling fibre pull-out and thus the total energy release rate.

Reactive fibre reinforced glass ionomer cements.

The mechanical properties of glass ionomer cements used in restorative dentistry reinforced by chopped glass fibres were investigated. Reactive glass fibres with a composition in the system SiO(2)-Al(2)O(3)-CaF(2)-Na(3)AlF(6) and a thickness of 26 microm were drawn by a bushing process. The manufacturing parameters were optimized with respect to maximum strength of the glass fibre reinforced ionomer cements. Powder to liquid ratio, pre-treatment of the glass, grain size distribution and fibre volume fraction were varied. Glass fibre and cement were characterized by X-ray diffraction, transmission electron microscopy and energy dispersive spectroscopy techniques, respectively. The highest flexural strength of the reinforced cement (15.6 MPa) was found by compounding 20 vol% reactive fibres and extending the initial dry gelation period up to 30 min. Microscopic examination of the fractured cements indicated a distinct reactive layer at the fibre surface. A pronounced fibre pull out mode gives rise to an additional work-of-fracture contributed by pulling the fibres out of the fracture surface.