Dynamic structures of motor proteins myosin and kinesin, and switch protein troponin as detected by SDSL-ESR

We have studied biological nano-machines, motor and switch proteins operating as supramolecular complexes by electron spin resonance (ESR) and found key features of their molecular movements. In all the systems, the specific movements of elements or domains were detected and quite dynamic at nanometer scale. We have observed two broad but distinct orientations, separated by a 25 degrees axial rotation, of a spin label attached specifically to the light chain (LC) domain of myosin motor in the muscle fibers. The distribution became only narrower upon muscle activation. ESR spectrum from the spin label of the neck-linker of dimeric kinesin motor consisted of immobilized and mobilized components and did not exhibit nucleotide-dependent mobility change. The distance between two labels of kinesin dimer was also measured by spin dipole-dipole interaction, showing a broad distribution and a nucleotide-dependent change on the nanometer scale (>1.5 nm). These results suggest that two LC domains of myosin and two neck linkers of kinesin play a similar role for sliding movement using two conformations. The spin label of the skeletal (Tn)-I regulatory domain (TnIreg) showed a large mobility change by Ca2+ ion suggesting a Ca-induced switch movement of TnIreg. Spin dipole-dipole interaction showed that in reconstituted muscle fibers both skeletal and cardiac TnC undergo Ca2+-induced structural change that is thought to be essential for TnIreg movement. We also succeeded in fixing the newly-synthesized bifunctional spin label rigidly on the TnC molecule in solution, indicating that we can determine the precise coordinate of the spin principal axis of troponin on the oriented filament.     

Hot pressing of SiC-TiC composites

Sintering behaviour during hot pressing of SiC-TiC composite ceramics has been investigated with special emphasis on the effect of various processing parameters on the density and mechanical properties of the sintered body. At hot pressing temperatures greater than 2000° C, significant densification occurred in SiC-50 wt%TiC (–0.5 wt%B-1 wt%C) composites. The room temperature flexural strength of the sintered body increased with the hot pressing temperature up to 2000° C and reached a highest value of 710 MPa in accordance to the variation of density with temperature. In sintering of composites without additives, densification was enhanced with the addition of up to 25 wt%TiC, with relative densities higher than 98% observed when hot pressing at 2150° C for 2h.     

Measurement of electron beam broadening in stainless steels during EDS analysis in the FEG-TEM

The effect of electron beam broadening on spatial resolution of EDS analysis in a 200 kV cold-FEG-TEM has been investigated for Type 316L stainless steel. Platinum (Pt) was evaporated on half the 316L TEM foil surface, and the difference in Pt concentration profiles obtained by the EDS method from the Pt evaporated surface and its opposite side was analysed as a function of foil thickness. Advantages of Pt use are that Pt is not included in the 316L steel, and it is easily formed as a stable and continuous thin evaporated film on the stainless steel. No change of spatial resolution caused by electron beam broadening was observed in stainless steel foils of less than about 140 nm thick subjected to EDS analysis with the FEG-TEM (incident probe size 1.6 +/- 0.2 nm diameter). Specimens of about 160 nm and more thick had a decrease in spatial resolution, but the decrease was smaller than that predicted from theoretical calculations of beam broadening for stainless steels. It is suggested that only the central part of the broadened electron beam with high intensity influences the spatial resolution of EDS analysis with the FEG-TEM.     

Evaluation of contribution for voltage control ancillary services based on social surplus

Reactive power supply plays an important role in active power supply with adequate system voltages. Various pricing mechanisms for reactive power supply have been developed and some of them have been adopted in some power systems, but they are in a trial stage. The authors also focus on development of a pricing method for reactive power ancillary services. This problem involves two technical issues: rational estimation of the cost associated with reactive power supply, and fair and transparent allocation of the estimated cost among the market participants. This paper proposes methods for evaluating the contribution of generators and demands. © 2010 Wiley Periodicals, Inc. Electr Eng Jpn, 174(2): 36–45, 2011; Published online in Wiley Online Library ( wileyonlinelibrary.com ). DOI 10.1002/eej.21016     

Constitutive production of aconitate isomerase by Pseudomonas sp. WU-0701 in relation to trans-aconitic acid assimilation

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Structural aspects of the LCST phase behavior of poly(benzyl methacrylate) in room-temperature ionic liquid

The structure and lower critical solution temperature (LCST) phase behavior of well-defined poly(benzyl methacrylate) (PBnMA) solution using 1-ethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)amide, [C₂mim][NTf₂] ionic liquid (IL) as a solvent have been studied by dynamic light scattering (DLS) and small-angle neutron scattering (SANS) at various temperatures. The SANS profiles observed for fully deuterated IL ([C₂mim]-d₁₁[NTf₂]) containing PBnMA were kept practically unchanged in the temperature range between 298 and 363 K, while they suddenly changed at 363 K. This indicates that the LCST behavior of PBnMA-IL solution is a first-order phase transition, which is consistent with the DLS results. The SANS profiles below 363 K were well represented by the theoretical Debye scattering function with inter-molecular interaction and the radius of gyration, R_g was estimated to be almost constant, i.e., ∼45 Å. The SANS result obtained here was compared with those in aqueous PNIPAm solutions as a typical LCST system, and some differences between IL and aqueous solution systems are pointed out. It is found that thermodynamic quantities (ΔH_demix, ΔS_demix and ΔG_demix) from the homogeneous solution to the phase separation states strongly depend on the solvation of the PBnMA polymer by the IL ([C₂mim] cation and [NTf₂] anion). We propose an LCST phase separation mechanism in the polymer-IL solution.     

A New Indicator to Differentiate Thyroid Follicular Inclusions in Cervical Lymph Nodes from Patients with Thyroid Cancer

Nodal metastasis is crucial for determining the stage of well-differentiated thyroid cancer (WTC) in patients older than 55. Well-formed thyroid follicular inclusions (TFIs) are occasionally encountered in the cervical lymph nodes (LNs) of patients with papillary thyroid carcinoma (PTC), and it is difficult to determine whether they are true nodal metastases or ectopic thyroid tissues (ETT). This study aimed to elucidate the impact of the expression of the DNA damage response molecule TP53-binding protein 1 (53BP1) using immunofluorescence (IF) as a biomarker to differentiate TFIs in cervical LN by comparing the mutation analyses of primary thyroid cancers. The data demonstrated the necessity for the differential diagnosis of true metastases from ETT among TFIs in cervical LNs. PTC-like nuclear features using hematoxylin–eosin staining combined with immunohistochemistry for conventional biomarkers of PTC, including BRAF^V600E protein, were most helpful in identifying metastatic follicular-patterned carcinomas. In conclusion, IF analysis of 53BP1 expression could be an excellent ancillary technique to distinguish metastatic carcinoma or ETT from TFIs in LNs, particularly in cases other than BRAF^V600E-mutated PTC.