Properties and substrate specificities of proteolytic enzymes from the edible basidiomycete Grifola frondosa

Highly active proteolytic enzymes are found in the fruiting bodies of Grifola frondosa. The general properties and substrate specificities of these proteases from G. frondosa (ProGF) were studied. The optimal pH for ProGF activity was pH 3 or 7 using hemoglobin or Hammersten casein as a substrate, respectively. The ProGF exhibited over 70% of maximal activity within the pH range of 4.5–8.5. In terms of temperature, the ProGF were maximally active at 55 °C, while over 80% of maximal activity was observed within the range of 50–75 °C. These proteases were substrate-specific, mainly cleaving at Ala¹⁴-Leu¹⁵, Tyr¹⁶-Leu¹⁷, and Pro²⁸-Lys²⁹ bonds, with occasional cleavage of Phe²⁴-Phe²⁵ bonds in the oxidized insulin B-chain. The ProGF also liberated hydrophobic amino acids, such as valine, leucine, and phenylalanine, using the oxidized insulin B-chain as a substrate. When soy protein was used as a substrate, valine, leucine, phenylalanine, and tyrosine were selectively released from the hydrolysate. Thus, over the time course of incubation, the peptide concentration increased as the average peptide chain length decreased. These results indicate that the ProGF include both endopeptidases recognizing leucine, phenylalanine, and lysine at the P1′ position, and aminopeptidases preferentially releasing hydrophobic and aromatic amino acids such as valine, leucine, phenylalanine, and tyrosine.     

Steroid hormone profiles of urban and tidal rivers using LC/MS/MS equipped with electrospray ionization and atmospheric pressure photoionization sources

A highly sensitive and uncomplicated method of analyzing steroidal hormones in river and estuarine water samples was developed using a liquid chromatography tandem mass spectrometer equipped with an electrospray ionization (ESI) source and atmospheric pressure photoionization (APPI) source. Steroidal hormones included not only estrogen but also androgen and conjugates of these two. APPI displayed greater sensitivity than ESI for most of the unconjugated steroids examined, with very high sensitivity for testosterone and 4-androstene-3,17-dione in particular. For conjugated hormones, in contrast, ESI was more effective. The method developed was applied to the determination of hormones in the rivers of Osaka City and their estuaries, where the hormones detected were affected by the effluent from municipal wastewater treatment plants (WWTPs), and hormone concentration values were comparable to those reported in previous studies of such effluent. Because of the two-way flow and stagnancy of streams and watercourses, continuous input of steroidal hormones from WWTPs seems to bring about local accumulation. Levels of androgen were 1 order of magnitude lower than those of estrogen. Estrone, estrone 3-sulfate, and 4-androstene-3,17-dione were detected in almost all water samples, with maxima of 51, 5.1, and 6.4 ng L(-1), respectively.     

LSI implementation of a low-power 4×4-bit array two-phase clocked adiabatic static CMOS logic multiplier

As the density and operating speed of complementary metal oxide semiconductor (CMOS) circuits increases, dynamic power dissipation has become a critical concern in the design and development—of personal information systems and large computers. The reduction of supply voltage, node capacitance, and switching activity are common approaches used in conventional CMOS. In adiabatic switching circuits, the current flow through transistors can be significantly reduced by ensuring uniform charge transfer over the entire available time. This paper presents the simulation of this current in two-phase clocked adiabatic static CMOS logic (2PASCL) and conventional CMOS. From the SPICE simulations, at transition frequencies from 1 to 12 MHz, a 4×4-bit array 2PASCL multiplier shows a maximum reduction in power dissipation of 77% relative to that of a static CMOS. The measurement results of a 4×4-bit array 2PASCL multiplier demonstrate a 57% reduction compared to a 4×4-bit array two-phase clocked adiabatic dynamic CMOS logic (2PADCL). These results indicate that 2PASCL technology can be advantageous when applied to low-power digital devices operated at low frequencies, such as radio-frequency identification (RFID) tags, smart cards, and sensors.     

Rheological characterization of konjac glucomannan in concentrated solutions

Dynamic viscoelasticity measurements were performed for concentrated solutions of konjac glucomannan in an ionic liquid. The entanglement coupling appeared in the rheological data for each solution was characterized in terms of the molecular weight between entanglements (M _e) as an average size of the transient entanglement network. The value of M _e for konjac glucomannan in the molten state was estimated to be 1.8 × 10³ (in g mol^?1), being significantly smaller than that for cellulose, although the molecular weight and linkage of the repeating units were the same between these polysaccharides. This result suggested that the configuration of the repeating monosaccharide unit affected the entanglement network of these polysaccharides reflecting the single chain characteristics.     

PATTERNED COPPER PLATING LAYER THICKNESS MADE UNIFORM BY PLACEMENT OF AUXILIARY GRID ELECTRODE ABOUT BALL GRID ARRAYS

The authors established a method to ensure uniform patterned copper plating layer thickness in semiconductor packages with high-density electrical circuits or ball grid arrays (BGA). A single BGA pattern unit was divided into small areas and for each, active area density was defined according to the area covered by the copper layer. Current distribution was determined for such an area by numerically solving Laplace and Butler-Voltmer equations, with consideration directed to active area density and plating solution conductivity. With the auxiliary grid electrode placed at a certain distance from the BGA so as to surround the latter by the former electrode, the current was found to be quite effectively prevented from becoming concentrated on sparsely dispersed circuit patterns generally situated near the edge of a unit BGA. Current distribution was noted to vary according to position and cross section size. Error could be made as little as 3.37% when this position was optimal.     

Double-shot inkjet printing of donor-acceptor-type organic charge-transfer complexes: Wet/nonwet definition and its use for contact engineering

We developed the double-shot inkjet printing technique for manufacturing the uniform polycrystalline thin films of organic charge-transfer complexes with a well-defined patterning on the substrate surfaces. The technique utilizes the wet/nonwet surface modification to confine the intermixed droplets of individually-printed donor and acceptor inks in a predefined area, which results in the picoliter-scale instantaneous complex formation. By the method, we obtained synthetic-metal patterned deposits of tetrathiafulvalene-tetracyanoquinodimethane with smooth thickness distribution and highly c-axis orientation, where the preference for the 1:1 complex formation is clearly demonstrated. We also discuss the use of the patterned deposits for the contact engineering of organic thin-film transistors.     

An investigation of the effect of fatigue deformation on the residual mechanical properties of Ti-6Al-4V ELI

Tensile properties, hardness, and Charpy impact toughness of Ti-6Al-4V extralow interstitial (ELI) with equiaxed α and Widmanstätten α structures at various stages of fatigue were investigated. Fatigue crack initiation characteristics of the same alloy were also investigated in this study. In the equiaxed α structure, fatigue cracks initiated mainly at the interface between primary-α grains, while in the Widmanstätten α structure, they initiated across α plates at an angle of around 45 deg to the stress axis. Specimens with the Widmanstätten α structure fractured before adequate fatigue hardening was achieved because a multitude of microcracks readily formed. Specimens with the equiaxed α structure fractured after adequate fatigue hardening developed. Tensile strength, 0.2 pct proof stress, and hardness increased clearly with increasing stress cycles and fatigue steps, particulary in the low-cycle fatigue (LCF) region, while impact toughness and elongation showed a reverse trend. It is suggested, therefore, that the dislocation density multiplies more rapidly near the specimen surface during the early stages of fatigue, while during the later stages of fatigue, dislocation density increases near the center of the specimen. Also, the dislocation multiplication will continue until saturation of the entire specimen has occurred.     

Effects of microstructure on the short fatigue crack initiation and propagation characteristics of biomedical α/β titanium alloys

This article presents the results of a study of the effects of microstructure on the fatigue strength and the short fatigue crack initiation and propagation characteristics of a biomedical α/β titanium alloy, Ti-6Al-7Nb. The results are compared to those obtained from a Ti-6Al-4V extra-low interstitial (ELI) alloy. Fatigue crack initiation occurs mainly at primary α grain boundaries in an equiaxed α structure, whereas, in a Widmanst?tten α structure, initiation occurs within the α colonies and prior β grains, where α plates are inclined at around 45 deg to the stress-axis direction. In an equiaxed α structure, the short fatigue crack initiation and propagation life, where the length of the crack (a) is in a microstructurally short fatigue-crack regime (2a < 50 μm), occupies around 50 pct of the total fatigue life. On the other hand, the fatigue crack in a Widmanst?tten α structure initiates at very early stages of fatigue, and, therefore, the fatigue crack-initiation life occupies a few percentages of the total fatigue life in an α structure. Then, the short fatigue crack propagates rapidly and is arrested at the grain boundaries of α colonies or prior β grains for a relatively long period, until the short crack passes through the boundaries to specimen failure. Therefore, the short fatigue crack-arrest life occupies more than 90 pct of the total fatigue life in a Widmanst?tten α structure. These trends are similar between the Ti-6Al-7Nb and Ti-6Al-4V ELI alloys and biomedical α/β titanium alloys. The total fatigue life for the Ti-6Al-7Nb alloy with an equiaxed α structure is changed by the volume fraction of primary α phase and the cooling rate after solution treatment. By increasing the volume fraction of the primary α phase from 0 to 70 pct, the fatigue limit of the Ti-6Al-7Nb alloy is raised. Changing the cooling rate after solution treatment by switching from air cooling to water quenching improves the fatigue limit of the Ti-6Al-7Nb alloy significantly.