Expression and circular dichroism studies of the extracellular domain of the alpha subunit of the nicotinic acetylcholine receptor

To provide material suitable for structural studies of the nicotinic acetylcholine receptor, we have expressed and purified the NH2-terminal extracellular domain of the mouse muscle alpha subunit. Several constructs were initially investigated using Xenopus oocytes as a convenient small scale expression system. A fusion protein (alpha210GPI) consisting of the 210 NH2-terminal amino acids of the alpha subunit and a glycosylphosphatidylinositol anchorage sequence conferred surface alpha-bungarotoxin binding in oocytes. Coexpression of alpha210GPI with an analogous construct made from the delta subunit showed no evidence of heterodimer formation. The alpha210GPI protein was chosen for large scale expression in transfected Chinese hamster ovary cells. The alpha210GPI protein was cleaved from these cells and purified on an immunoaffinity column. Gel and column chromatography show that the purified protein is processed as expected and exists as a monomer. The purified protein also retains the two distinct, conformation-specific binding sites expected for the correctly folded alpha subunit. Circular dichroism studies of alpha210GPI suggest that this region of the receptor includes considerable beta-sheet secondary structure, with a small proportion of alpha-helix.     

The mouse embryo culture system: improving the sensitivity for use as a quality control assay for human in vitro fertilization

Endurance training leads to an increase in the content of individual mitochondrial hemeproteins in skeletal muscle. To deduce the control mechanisms involved, cytochrome oxidase (COX) activity was compared with 1) the content of COX subunits III and IV and 2) 5-aminolevulinate synthase (ALAS) activity and mRNA content. In the plantaris muscle of female rats run daily for up to 28 days, ALAS activity was elevated 100% (P < 0.01) after 3 days and remained 150 and 125% higher (P < 0.001) after 7 and 28 days of running, respectively, than control. COX activity was also increased, but not until day 7 (40%; P < 0.05), and reached a maximal value 80% higher than control (P < 0.001) in the 28-day group. Compared with control, the content of COX subunit III and IV and ALAS mRNAs was not significantly changed by the training. The increased activities of COX and ALAS appear to be regulated by translational or posttranslational steps in the protein expression pathway. The induction of ALAS before COX suggests that the increased activity of COX may require increased synthesis of heme.     

Effect of Dilution and Porosity on Self-Propagating High-Temperature Synthesis of Silicon Nitride

Self-propagating high-temperature synthesis is a very easy and low-cost method to synthesize Si₃N₄. The nitriding of silicon powder takes place in a self-sustained regime under high pressures of nitrogen with dilution of silicon by Si₃N₄. In this work effects of dilution and green-mixture porosity on combustion velocity and phase content of reaction products are studied. Results are compared with previous work of other authors and different behaviors are found. An explanation of these behaviors is given.     

Inside Front Cover: Three‐Dimensionally Ordered Gold Nanocrystal/Silica Superlattice Thin Films Synthesized via Sol–Gel Self‐Assembly (Adv. Funct. Mater. 7/2006)

The synthesis of three‐dimensionally ordered, transparent gold‐nanocrystal (NC)/silica superlattice thin films using the self‐assembly (by spin‐coating) of water‐soluble gold nanocrystal micelles and soluble silica is reported by Fan and co‐workers on p. 891. The robust, 3D NC/silica superlattice films are of interest for the development of collective optical and electronic phenomena, and, importantly, for the integration of NC arrays into device architectures. Nanocrystals and their ordered arrays hold many important applications in fields such as catalysis, surface‐enhanced Raman spectroscopy based sensors, memory storage, and electronic and optical nanodevices. Herein, a simple and general method to synthesize ordered, three‐dimensional, transparent gold nanocrystal/silica superlattice thin films by self‐assembly of gold nanocrystal micelles with silica or organosilsesquioxane by spin‐coating is reported. The self‐assembly process is conducted under acidic sol–gel conditions (ca. pH 2), ensuring spin‐solution homogeneity and stability and facilitating the formation of ordered and transparent gold nanocrystal/silica films. The monodisperse nanocrystals are organized within inorganic host matrices as a face‐centered cubic mesostructure, and characterized by transmission electron spectroscopy and X‐ray diffraction.     

Grain size dependence of the activation parameters for plastic deformation: Influence of crystal structure,slip system,and rate-controlling dislocation mechanism

A critical review of available results on the dependence of grain size on the activation parameters for deformation, specifically, the activation volume, V*, and the thermal component of flow stress, σ*, has been carried out with a view to verifying the Armstrong prediction that identifies the Hall-Petch (H-P) intercept with the easy slip system and the H-P slope with the most difficult system in polycrystals. The influence of slip system choice is demonstrated using results on Cd and Zr. The Armstrong prediction is valid for basal slip hcp metals, such as Cd and Zn, with V* and σ* determined by the difficult pyramidal slip. For the prism slip metals such as Zr and Ti, V* and σ* are controlled by interstitial solutes and are independent of grain size. The results on Zr are used to highlight the influence of dynamic strain aging on the H-P parameters. In bcc metals, in which the Peierls-Nabarro barrier is the rate-controlling obstacle, V* and σ* are again independent of grain size. For fcc metals, correlation of the H-P slope with the cross-slip stress, predicted by the Armstrong model, has been demonstrated for a few cases. The variation of V* with grain size in Ni as reported by Narutani and Takamura (Acta Metall. Mater., 1991, vol. 227, pp. 2037–49) is newly interpreted in terms of the Armstrong model that associates the H-P intercept in fcc metals with dislocation intersections and the H-P slope with cross-slip, and provides realistic results for the activation volumes for the two processes. This article is based on a presentation given in the symposium “Dynamic Deformation: Constitutive Modeling, Grain Size, and Other Effects: In Honor of Prof. Ronald W. Armstrong,” March 2–6, 2003, at the 2003 TMS/ASM Annual Meeting, San Diego, California, under the auspices of the TMS/ASM Joint Mechanical Behavior of Materials Committee.