Ceruloplasmin-induced aggregation of P19 neurons involves a serine protease activity and is accompanied by reelin cleavage.

The cytoarchitectural organization of the nervous system depends partly on extracellular serine proteases, including reelin. This 400K protein, which also exists as the N-terminally-derived 300K and 180K fragments, acts through binding to the lipoprotein receptors apolipoprotein E receptor 2 (ApoER2) and very low-density lipoprotein receptor (VLDLR). Ceruloplasmin (CP), a multifunctional protein found in the circulation and also expressed on glial cells, was shown to bind to, and induce aggregation of neurons newly differentiated from P19 embryonic stem cells. This indicated a potential developmental role of CP in neuronal organization, possibly in relation with reelin and other extracellular serine proteases. Therefore, we analysed the effect of cell-impermeant, large spectrum, serine protease inhibitors on CP-induced neuroaggregation and studied reelin expression. Soybean trypsin inhibitor and aprotinin (SBTI+Apro) inhibited CP neuroaggregative action. Undifferentiated and neurally-differentiating cultures secreted the 400K reelin. The 180K fragment was present during and after differentiation whereas the 300K species was barely detectable. However, CP stimulated generation of the 300K in the differentiated neuronal cultures, and SBTI+Apro abolished this CP effect. Time course profiles and function-blocking antibody indicated that neuroaggregation does not depend on the generation of the 300K fragment or on reelin action. CP neuroaggregative action thus involves a pericellular serine protease, different from reelin. On the other hand, the CP stimulation of reelin cleavage is in line with a possible role of CP in nervous system development. Since P19 cells express ApoER2 and VLDLR, they can help understanding relationships existing between CP, reelin and intervening protease(s).

Ability of various impression materials to produce duplicate dies from successive impressions.

This in vitro study evaluated the ability of five different impression techniques to make duplicate dies of two different types of tooth preparation. One mandibular second premolar Ivorine tooth was prepared for a complete crown and one for an onlay. A master impression was made of each tooth preparation with the use of five impression techniques for a total of 10 master impressions, and a master die was made from each of these impressions. Castings were made on these master dies, and the fit of each casting was verified on the respective Ivorine tooth. Marginal openings of the castings on the master dies were recorded under magnification at four predetermined points. Five successive impressions, with the use of each impression material, were then made of each tooth preparation for a total of 50 test impressions, and 50 test dies were made from these impressions. The fit of the respective casting was evaluated under magnification for each test die at the four predetermined points, and marginal openings were recorded. Differences between the marginal discrepancies of the casting on the master die and on the test die were tabulated and the results were statistically analyzed. Results indicated that none of the impression materials was capable of producing exact replicas. Polysulfide rubber performed significantly better than two materials for the production of duplicate dies with the complete crown preparation; and polyvinyl siloxane used with a putty-light body, single-stage technique produced mean marginal discrepancies that were significantly greater than the other four techniques when used for the onlay preparation.(ABSTRACT TRUNCATED AT 250 WORDS)