Recent evidence from genetic experiments in yeast and from studies using guanosine triphosphate (GTP) analogues in mammalian cells suggests a key role for low-molecular-mass GTP-binding proteins (LMM-GBPs) (Mr 19 to 28 kD) in processes of intracellular vesicular sorting and secretion. Assembly and exocytosis of the lamellar body (LB), the secretory organelle of the pulmonary alveolar type 2 pneumocyte, may be regulated by LMM-GBPs. We used [alpha-32P]GTP binding to Western blotted proteins, ultraviolet crosslinking of [alpha-32P]GTP to membrane proteins, immunoblotting with specific antisera, and botulinum exoenzyme C3-catalyzed ADP ribosylation to detect LMM-GBPs in LB. With the first two techniques, we have identified six LMM-GBPs of approximately 27, 25.5, 24.5, 23, 22, and 21 kD that are enriched in a highly purified LB fraction compared with type 2 pneumocyte homogenate, crude membranes, and cytosol. Further characterization of the LB LMM-GBPs by immunoblotting revealed that ras p21 is greatly enriched in the LB fraction compared with other type 2 pneumocyte fractions. In addition, botulinum exoenzyme C3 catalyzed the ADP ribosylation of 20- to 21-kD proteins that were similarly enriched in the LB fraction. In contrast, a monospecific antibody to ADP-ribosylation factor reacted with a 19-kD protein only in the type 2 pneumocyte homogenate and cytosol fractions. Monospecific antibodies to yeast Sec4 protein and to rab 3A did not react with any type 2 pneumocyte proteins. The LMM-GBPs specifically associated with LB may participate in intracellular events required for surfactant packaging and secretion.

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This paper demonstrates developments that introduce generalized Bezier components in the Moving Morphable Components (MMC) optimization framework. Methods of enhancing the parameterization of the components to provide the opportunity for a better optimum, than can be achieved using existing approaches, are also described. The use of control points and Bezier curves for representing structural components provides both additional flexibility in the shape and a parameterization that complies with extrude and swept feature-based templates available in commercial computer-aided design (CAD) packages. Methods of representing these structural components, calculating analytical derivatives, and numerical examples demonstrating their integration in the MMC framework, are presented for a series of author-derived and literature problems. A successive refinement technique demonstrates how the additional flexibility in the structural components enables progressive improvement in the objective function. For the examined problems, increasing the design variables per component (from 5 to 15) resulted in solutions with 6% to 36% reduction in compliance. This improvement was achieved without increasing the number of components in the design space.