Effective Characterization of DNA Ligation Kinetics by High-Resolution Melting Analysis

High-resolution melting (HRM) analysis has been improved and applied for the first time to quantitative analysis of enzymatic reactions. By using the relative ratios of peak intensities of substrates and products, the quantitativity of conventional HRM analysis has been improved to allow detailed kinetic analysis. As an example, the ligation of sticky ends through the action of T4 DNA ligase has been kinetically analyzed, with comprehensive data on substrate specificity and other properties having been obtained. For the first time, the kinetic parameters (k_obs and apparent K_m) of sticky-end ligation were obtained for both fully matched and mismatched sticky ends. The effect of ATP concentration on sticky-end ligation was also investigated. The improved HRM method should also be applicable to versatile DNA-transforming enzymes, because the only requirement is that the products have T_m values different enough from the substrates.     

Development of Selective TGR5 Ligands Based on the 5,6,7,8-Tetrahydro-5,5,8,8-tetramethylnaphthalene Skeleton

TGR5, a G-protein-coupled receptor (GPCR), plays an important role in several physiological functions. TGR5 activation through bile acids induces an increase in energy expenditure. Therefore, synthetic TGR5 ligands could be useful for the treatment of obesity or dyslipidemia. In this study, we designed and synthesized a set of TGR5 ligands with a 5,6,7,8-tetrahydro-5,5,8,8-tetramethylnaphthalene (TMN) skeleton, and evaluated their TGR5 agonistic activity. We also investigated the selectivity of the synthesized compounds for TGR5 relative to the farnesoid X receptor (FXR) and retinoic acid receptor (RAR). Our results show that compound 4 b [N-(2-chlorophenyl)-5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenecarboxamide] exhibited potent TGR5 agonist activity with an IC₅₀ value of 8.4 nM without significant cytotoxicity. In addition, compound 4 b showed only slight agonistic activity toward FXR and RAR at 1 μM treatment. These data indicate that compound 4 b is a selective TGR5 agonist, and could be a promising therapeutic agent for dyslipidemia.     

Aerobic Exercise Ameliorates Cancer Cachexia-Induced Muscle Wasting through Adiponectin Signaling

Cachexia is a multifactorial syndrome characterized by muscle loss that cannot be reversed by conventional nutritional support. To uncover the molecular basis underlying the onset of cancer cachectic muscle wasting and establish an effective intervention against muscle loss, we used a cancer cachectic mouse model and examined the effects of aerobic exercise. Aerobic exercise successfully suppressed muscle atrophy and activated adiponectin signaling. Next, a cellular model for cancer cachectic muscle atrophy using C2C12 myotubes was prepared by treating myotubes with a conditioned medium from a culture of colon-26 cancer cells. Treatment of the atrophic myotubes with recombinant adiponectin was protective against the thinning of cells through the increased production of p-mTOR and suppression of LC3-II. Altogether, these findings suggest that the activation of adiponectin signaling could be part of the molecular mechanisms by which aerobic exercise ameliorates cancer cachexia-induced muscle wasting.     

Improvement in impact strength of modified cardanol‐bonded cellulose thermoplastic resin by adding modified silicones

The impact strength of cellulose diacetate (CDA) bonded with a modified cardanol (3‐pentadecylphenoxy acetic acid: PAA) was greatly improved up to 9 kJ/m² by adding a relatively small amount of modified silicones while suppressing a decrease in bending strength. In our recent research, this thermoplastic resin (PAA‐bonded CDA) exhibited high rigidity, glass transition temperature, and water resistance. However, its impact strength was insufficient for use in durable products. Therefore, silicones modified with polyether, amino, and epoxy groups were investigated as possible ways to improve the impact strength. The results show that adding polyether‐modified silicone (polyether silicone) with moderate polarity relative to PAA‐bonded CDA resulted in shearing deformation greatly enhances its impact strength while maintaining other properties, including glass transition temperature (T_g), water resistance, and thermoplasticity. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40366.     

Improvement in impact strength of modified cardanol‐bonded cellulose thermoplastic resin by using olefin resins

Impact strength of a modified cardanol‐bonded cellulose thermoplastic resin was greatly improved by using a small amount of olefin resins. As we showed, this thermoplastic resin (3‐pentadecylphenoxy acetic acid (PAA)‐bonded cellulose diacetate (CDA): PAA‐bonded CDA) exhibited high practical properties such as bending strength, heat resistance, and water resistance. However, its impact strength was insufficient for use in durable products. We improved the impact strength of PAA‐bonded CDA by adding hydrophobic olefin resins, such as polyethylene or polypropylene, while maintaining good bending strength and breaking strain. Furthermore, the application of olefin resins also increased water resistance and fluidity. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 39829.     

Mechanical role of reinforcement in seismic behavior of steel-strip reinforced earth wall

In the current design practices of steel-strip reinforced earth walls (SSREWs), the length of the reinforcing material is determined based on the equilibrium between the reinforcement tension and the earth pressure acting on the wall. Here, the resistance of the reinforcing material laid in the active failure zone (AFZ) is not considered. Moreover, the mechanical role of the reinforcing material against the integrity of the SSREW has not been sufficiently verified. Regarding the seismic stability of SSREW, although it is investigated by treating the entire reinforced earth wall as a rigid body, this inspection method is for gravity-retaining walls, and the difference in the seismic behavior between the SSREW and the rigid body is not clear. In this study, therefore, dynamic centrifuge model tests on 6 types of SSREWs were conducted to clarify the following items: (1) the basic earthquake behavior of a SSREW, (2) the mechanical role of the reinforcing material laid in the AFZ and (3) the mechanical role of the reinforcing material against the integrity of the SSREW. The results indicated that the reinforcing material laid in the AFZ can restrain the amount of deformation of the wall during earthquakes. Furthermore, the more stable the AFZ is, the smaller the maximum wall displacement will be.     

Terpendole E and its Derivative Inhibit STLC‐ and GSK‐1‐Resistant Eg5

Terpendole E is first natural product found to inhibit mitotic kinesin Eg5, but its inhibitory mechanism remains to be revealed. Here, we report the effects of terpendole E and 11ketopaspaline (a new natural terpendole E analogue) on the Eg5–microtubule interaction and in several Eg5 mutants. 11‐Ketopaspaline is a shunt product from terpendole E, and it shows potent inhibitory activity against the microtubule‐stimulated ATPase activity of Eg5. Unlike other Eg5 inhibitors, such as S‐trityl‐L ‐cysteine (STLC) and GSK‐1, both terpendole E and 11‐ketopaspaline only partially inhibited Eg5–microtubule interaction. Furthermore, terpendole E and 11‐ketopaspaline inhibited several Eg5 mutants that are resistant to STLC (Eg5^D130A, Eg5^L214A) or GSK‐1 (Eg5^I299F, Eg5^A356T), but with the same extent of inhibition against wild‐type Eg5. Because Eg5^D130A and Eg5^L214A show cross‐resistance to most known Eg5 inhibitors, which bind the L5 loop, these results suggest that terpendole E and its analogues have a different binding site and/or inhibitory mechanism to those for L5 loop‐binding type Eg5 inhibitors.     

Investigation of hydrogen plasma treatment for reducing defects in silicon quantum dot superlattice structure with amorphous silicon carbide matrix

We investigate the effects of hydrogen plasma treatment (HPT) on the properties of silicon quantum dot superlattice films. Hydrogen introduced in the films efficiently passivates silicon and carbon dangling bonds at a treatment temperature of approximately 400°C. The total dangling bond density decreases from 1.1 × 10¹⁹ cm^-3 to 3.7 × 10¹⁷ cm^-3, which is comparable to the defect density of typical hydrogenated amorphous silicon carbide films. A damaged layer is found to form on the surface by HPT; this layer can be easily removed by reactive ion etching.     

Silicon quantum dot superlattice solar cell structure including silicon nanocrystals in a photogeneration layer

The solar cell structure of n-type poly-silicon/5-nm-diameter silicon nanocrystals embedded in an amorphous silicon oxycarbide matrix (30 layers)/p-type hydrogenated amorphous silicon/Al electrode was fabricated on a quartz substrate. An open-circuit voltage and a fill factor of 518 mV and 0.51 in the solar cell were obtained, respectively. The absorption edge of the solar cell was 1.49 eV, which corresponds to the optical bandgap of the silicon nanocrystal materials, suggesting that it is possible to fabricate the solar cells with silicon nanocrystal materials, whose bandgaps are wider than that of crystalline silicon.

PACS

85.35.Be; 84.60.Jt; 78.67.Bf