High Resolution Structure of the Mature Capsid of Ralstonia solanacearum Bacteriophage ϕRSA1 by Cryo-Electron Microscopy

The ϕRSA1 bacteriophage has been isolated from Ralstonia solanacearum, a gram negative bacteria having a significant economic impact on many important crops. We solved the three-dimensional structure of the ϕRSA1 mature capsid to 3.9 Å resolution by cryo-electron microscopy. The capsid shell, that contains the 39 kbp of dsDNA genome, has an icosahedral symmetry characterized by an unusual triangulation number of T = 7, dextro. The ϕRSA1 capsid is composed solely of the polymerization of the major capsid protein, gp8, which exhibits the typical “Johnson” fold first characterized in E. coli bacteriophage HK97. As opposed to the latter, the ϕRSA1 mature capsid is not stabilized by covalent crosslinking between its subunits, nor by the addition of a decoration protein. We further describe the molecular interactions occurring between the subunits of the ϕRSA1 capsid and their relationships with the other known bacteriophages.     

Implementation of vibration suppression control on FWL processor

In motion or process control systems, a variety of design techniques have been proposed because of the demand for high performance. The higher performance we demand, the higher the degree of the controller becomes. The controller is generally designed by a CAD system and implemented with a microprocessor. But the microprocessor does not have enough precision to realize the results of design by the CAD system. Therefore, the system performance is degraded by finite word length (FWL) effects. To deal with FWL problems, many design methods have been considered in the signal processing field, and high‐ordered digital filters are often used. Among these methods, the implementation technique based on the state‐space realization can minimize the sensitivity to perturbation of coefficients. Noting that optimal realizations with the same transfer function are unique only up to an orthogonal similarity transformation, we must choose the realization within this class of optimal realizations. In this paper, we present an algorithm to find a state‐space realization which minimizes the frequency‐weighted sensitivity measure of the controller performance. Furthermore, we present some experimental results to verify the effectiveness of the proposed algorithm. © 1999 Scripta Technica, Electr Eng Jpn, 128(1): 45–52, 1999     

Microstructural Changes During Plastic Deformation and Corrosion Properties of Biomedical Co-20Cr-15W-10Ni Alloy Heat-Treated at 873 K

Metallurgical and Materials Transactions A - Microstructural changes were observed during the plastic deformation of ASTM F90 Co-20Cr-15W-10Ni (mass pct) alloy heat-treated at...     

Effect of mutagenesis at the region upstream from the G(Q/E) motif of three types of geranylgeranyl diphosphate synthase on product chain-length

(All-E) geranylgeranyl diphosphate synthases have been classified into three types based on the characteristic sequences around the first aspartate rich motif, which is highly conserved among the enzymes. In type I geranylgeranyl diphosphate synthases, which consist of archaeal enzymes, a bulky amino acid residue at the 5th position upstream from the motif plays a main role in the product determination, by blocking further elongation of prenyl chain as the bottom of the reaction cavity. On the other hand, type III geranylgeranyl diphosphate synthases, which consist of the enzymes from eukaryotes except for plants, use a bulky amino acid residue at the 2nd position upstream from the conserved G(Q/E) motif for product chain-length determination. Thus we introduced mutations into the region upstream from the G(Q/E) motif of geranylgeranyl diphosphate synthases of the three different types to confirm the importance of the region for the product chain-length determination. The results of the mutational analyses indicated that not only the 2nd but also the 3rd position upstream from the G(Q/E) motif is involved in the product chain-length determination mechanism in types I and III geranylgeranyl diphosphate synthases, while the amino acid substitution in this region did not affect the chain-length of the products of type II geranylgeranyl diphosphate synthase, which consist of the enzymes from bacteria and plants. The region upstream from the G(Q/E) motif possibly contributes to the product determination in the wide range of geranylgeranyl diphosphate synthases, as well as that around the first aspartate rich motif.     

Improvement of biomolecule quantification precision and use of a single-element aspheric objective lens in fluorescence correlation spectroscopy

We found a way to increase the precision with which biomolecules present at concentrations below 10(-10) M can be quantified by fluorescence correlation spectroscopy (FCS). The effectiveness of the way was demonstrated experimentally by using a single-element aspheric objective lens, which was newly developed to reduce the cost of FCS instruments. In the first part of this paper, the relative standard deviation (RSD) of FCS-based concentration measurements is estimated theoretically by an analytical approximation assuming the detection volume profiles in FCS setups to be Gaussian and by molecular simulations in which more realistic profiles are calculated from physical parameters of the measurement setups. In a limit of infinitely bright molecules and zero background emission, the analytical approximation predicts that the RSD at a concentration is minimized when the mean number of molecules in a detection volume is approximately 0.5. A detection volume of the order of 10(-13) L thus gives smaller RSD values for concentrations from 10(-11) to 10(-10) M than does one of the order of 10(-15) L, which is widely used in FCS. This prediction is supported by the molecular simulations, taking into account the finite molecule brightness and background emission. In the second part of the paper, the RSD is evaluated experimentally with an FCS setup with a detection volume of 1.1 x 10(-13) L. The newly developed objective lens, serving as the bottom of the sample cell in this setup, has a large numerical aperture (0.9) without using immersion liquid. When a calibration line was made by 30-s FCS measurements of Cy3-labeled, 112-mer single-stranded DNA solutions, the RSD roughly agreed with the simulation result and was less than 0.1 for DNA concentrations from 2 x 10(-11) to 10(-10) M.     

Simultaneous B-M-mode scanning method for real-time full-range Fourier domain optical coherence tomography

High-speed complex full-range Fourier domain optical coherence tomography (FD-OCT) is demonstrated. In this FD-OCT, the phase modulation of a reference beam (M scan) and transversal scanning (B scan) are simultaneously performed. The Fourier transform method is applied along the direction of the B scan to reconstruct complex spectra, and the complex spectra comprise a full-range OCT image. Because of this simultaneous B-M-mode scan, the FD-OCT requires only a single A scan for each single transversal position to obtain a full-range FD-OCT image. A simple but slow version of the FD-OCT visualizes the cross section of a plastic plate. A modified fast version of this FD-OCT investigates a sweat duct in a finger pad in vivo and visualizes it with an acquisition time of 27 ms.