Site-directed mutagenesis of the putative active site residues of 3C proteinase of coxsackievirus B3: evidence of a functional relationship with trypsin-like serine proteinases

Picornavirus 3C proteinases (3C^pro) are cysteine proteinasesbut recent sequence analyses have shown that they are relatedto trypsin-like serine proteinases. Two models of 3C^pro structurehave been presented. Both models indicate that residues His40and Cysl47 are members of the catalytic triad but the modelsdiffer in the designation of the third member of the catalytictriad, which is assigned as either Glu71 or Asp85. To test theimportance of these four residues in the catalytic activityof 3C^pro of coxsackievirus B3, a member of the enterovirus subgroupof the picornavirus family, single amino acid substitutionswere introduced at each of the four sites. All of these mutationsresulted in the reduction or inactivation of autocatalytic cleavageof the 3C precursor protein expressed in Escherichia coli, suggestingthat all of these residues are essential for the proteolyticreaction. The substitution of Cysl47 with Ala abolished 3C^proactivity while the mutant in which Cysl47 was replaced withSer retained reduced proteolytic activity both in cis and intrans. Our results strongly support the proposal that Cysl47of 3C^pro functions as a nucleophile analogous to Serl95 of trypsin-likeserine proteinases.     

Retarding intermetallic compounds growth of Zn high-temperature solder and Cu substrate by trace element addition

Since the thickness of layers of intermetallic compounds (IMCs) at the joining interface has an apparent effect on the joint reliability, understanding the growth mechanisms of IMCs is important for die-attachment applications. In the present study, the interfacial reaction between elements (Ca, Mn, Cr, and Ti) added to Zn alloys and Cu substrate were investigated in detail, focusing on IMCs grown by isothermal aging at 150 °C. The reaction layers included two types of Cu–Zn IMCs i.e., γ-Cu₅Zn₈ and ε-CuZn₅ phases. The joining interface with trace elements containing Zn solder reduced the growth rate of the IMCs. In particular, the addition of 0.1 wt% Cr to pure Zn solder decreased the growth rate of the IMCs by approximately 50 %. The mechanism we propose assumes that a phase containing the small Cr atoms exists between the solder and ε-CuZn₅, which suppresses the diffusion of metal atoms. To summarize, the Cr additive showed beneficial effects in terms of suppressing the growth of IMCs during the solid-state isothermal aging.     

Fabrication of a microelectromechanical system mirror array and its drive electrodes for low electrical interference in wavelength‐selective switches

This article describes the fabrication of a microelectromechanical system (MEMS) mirror array and mirror‐drive electrodes with high‐aspect‐ratio gold walls that reduce electrical interference in a wavelength‐selective switch (WSS). The MEMS mirror array, in which a lot of closely spaced adjacent mirrors are electrostatically operated, can be fabricated with a high yield by encapsulating the mirrors with an organic film that protects them from process damage. The gold walls with a high‐aspect ratio are formed in the narrow space between adjacent mirror‐drive electrodes by using thick‐multilevel interconnection technology. Because of these walls between adjacent electrodes, each MEMS mirror operates with low electrical interference. © 2011 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc.     

Effect of 9,12-Octadecadiynoic Acid on Neurobehavioral Development in Caenorhabditis elegans

Human breast milk lipids have major beneficial effects: they promote infant early brain development, growth and health. To identify the relationship between human breast milk lipids and infant neurodevelopment, multivariate analyses that combined lipidomics and psychological Bayley-III scales evaluation were utilized. We identified that 9,12-octadecadiynoic acid has a significantly positive correlation with infant adaptive behavioral development, which is a crucial neurodevelopment to manage risk from environmental stress. To further clarify the biological function of 9,12-octadecadiynoic acid in regulating neurodevelopment, Caenorhabditis elegans (C. elegans) was used as a model to investigate the effect of 9,12-octadecadiynoic acid on neurobehavioral development. Supplementation with 9,12-octadecadiynoic acid from the L1 to L4 stage in larvae affected locomotive behaviors and foraging ability that were not socially interactive, implying that 9,12-octadecadiynoic acid is involved in regulating the serotonergic neuronal ability. We found that supplementary 0.1 μM 9,12-octadecadiynoic acid accelerated the locomotive ability and foraging ability via increasing the expression of serotonin transporter mod-1. Antioxidant defense genes, sod-1, sod-3 and cyp-35A2 are involved in 9,12-octadecadiynoic acid-induced motor neuronal activity. Nevertheless, supplementary 9,12-octadecadiynoic acid at concentrations above 1 μM significantly attenuated locomotive behaviors, foraging ability, serotonin synthesis, serotonin-related gene expressions and stress-related gene expression, resulting in the decreased longevity of worms in the experiment. In conclusion, our study demonstrates the biological function of 9,12-octadecadiynoic acid in governing adaptive behavioral development.     

Microwave Dielectric Study on Water Structure and Physical Properties of Aqueous Systems Using Time Domain Reflectometry with Flat-End Cells

Microwave dielectric measurements were performed in the frequency range from 1 mHz up to 30 GHz using a time domain reflectometry (TDR) method for emulsions and gels. Flat-end sample cells have been used in the TDR measurement to contact a small spot of the surface of those viscoelastic and solid samples without any destruction. Relaxation processes due to various water structures were observed for these aqueous systems. Relaxation parameters thus obtained offer information about these water structures and amounts. The relaxation strength obtained from the high frequency process due to free water can be an adequate measure of water content in spite of some ambiguities for different water structures in some materials. Comparisons of actual water contents in emulsion with those estimated from the relaxation strength indicate that water structure is affected by the interaction between water and micelle. Unfreezable water observed in DNA gel under the freezing point consists of bound water and a fraction of free water. Bound water molecules are still unfreezable to keep the double helical structure of DNA, when the fraction of free water is frozen at lower temperatures. These water structures determine physical properties of moist materials. TDR measuring technique with the flat-end cell is effective to investigate water structures in viscoelastic moist materials and to evaluate physical properties and structures of complex molecular systems.     

Molecular orbital study of pyrolytic carbons based on small cluster models

Molecular orbital calculations are applied to the Raman scattering and ESR of pyrolytic carbons on the basis of small cluster models. The E_2g and A_1g modes of C-C stretching vibrations of coronene, hexabenzocoronene, and circumcoronene, which belong to D_6h carbon clusters, are shown to appear around the 1550 cm⁻¹ and 1360 cm⁻¹ bands, respectively. The unpaired electrons observed in pyrolytic carbons are attributed to the bond-alternation defects on odd-numbered carbon clusters that are more easily mobile than those of trans-polyacetylene.