Previous experiments showed that activity of the ؊800-base pair MLC2slow promoter was 75-fold higher in the innervated soleus (SOL) compared with the noninnervated SOL muscles. Using in vivo DNA injection of MLC2slow promoter-luciferase constructs, the aim of this project was to identify regulatory sites and potential transcription factors important for slow nerve-dependent gene expression. Three sites within the proximal promoter (myocyte enhancer factor-2 (MEF2), E-box, and CACC box) were individually mutated, and the effect on luciferase expression was determined. There was no change in luciferase expression in the SOL and extensor digitorum longus (EDL) muscles when the E-box was mutated. In contrast, the MEF2 mutation resulted in a 30-fold decrease in expression in the innervated SOL muscles (10.3 versus 0.36 normalized relative light units (RLUs)). Transactivation of the MLC2slow promoter by overexpressing MEF2 was only seen in the innervated SOL (676,340 versus 2,225,957 RLUs; p < 0.01) with no effect in noninnervated SOL or EDL muscles. These findings suggest that the active MLC2slow promoter is sensitive to MEF2 levels, but MEF2 levels alone do not determine nerve-dependent expression. Mutation of the CACC box resulted in a significant up-regulation in the EDL muscles (0.23 versus 4.08 normalized RLUs). With the CACC box mutated, overexpression of MEF2 was sufficient to transactivate the MLC2slow promoter in noninnervated SOL muscles (27,536 versus 1,605,797 RLUs). Results from electrophoretic mobility shift and supershift assays confirm MEF2 protein binding to the MEF2 site and demonstrate specific binding to the CACC sequence. These results suggest a model for nerve-dependent regulation of the MLC2slow promoter in which derepression occurs through the CACC box followed by quantitative expression through enhanced MEF2 activation.

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As a part of a SEMATECH project to improve the cost performance of semiconductor facility fluid systems, cost models have been developed in Microsoft Excel for gases chemicals, and ultra pure water (UPW) systems. These cost models are designed to permit cost analysis associated with the acquisition, use, and maintenance of fluid systems as well as cost comparisons of various fluid supply and distribution methods using a consistent costing methodology. The system attributes to be input in the model include the flowrate, and maintenanceheliability data. The subsystem attributes to be input in the model include the capital costs, leased equipment costs, utility usage, labor requirements, and other operating costs. Based on these inputs, the total capital and operating costs are calculated for a fluid system and ten year cost analysis is performed. The output is reported in terms of cost per unit volume.

The existing electrical grid infrastructure is aging and based on century old systems and ap-proaches to electrical delivery. Microgrid technology oers many advancements and benets to thefuture of the electrical grid. This paper looks at the inner makings of microgrids and their imple-mentation. This paper examines various storage methods within microgrids, a key benet to theirutilization, as well as a brief look at various microgrid control functions and issues. Regulatoryissues are covered to address the potential wide spread expansion of microgrids and their ability tobe the cornerstone of the electrical grid in the future.