Construction of the plasmid PMEX8-HAK1 and random site-directed mutagenesis of human cytosolic adenylate kinase

The pMEX8-hAK1 vector was devised from the pAK plasmid (Kim J. H. et al., 1989, Protein Engineering 5, 379-386), which could directly express human adenylate kinase proteins without recombination and its single strand DNA could be withdrawn with helper phage for random site-directed mutagenesis. The conserved key residues at Lys21, Lys27, and Thr39 were engineered to obtain mutants for kinetic analysis. Three mutants were obtained as K21P, K27R, and T39S, their specific activities were strikingly reduced compared to those of wild type adenylate kinase. This pMEX8-hAK1 will be a powerful tool for site-directed mutagenesis to detect the substrate-enzyme interaction for human adenylate kinase including various other enzymes.     

pH-controlled uphill transport of ammonium ions through polymer membranes with sulfonic acid groups

Uphill transport of ammonium ions through a membrane with sulfonic acid groups were investigated with pH-controllers which keep the solution at a constant pH. A membrane with sulfonic acid groups was prepared by casting an aqueous solution containing poly(styrenesulfonic acid) and poly(vinyl alcohol) on a glass plate. When a membrane with one side alkaline and the other acidic was fixed as a diaphragm in a cell, ammonium ions were transported from the alkaline side to the acidic side through the membrane against the concentration gradient of the adjacent solutions. Uphill transport of ammonium ions with pH-controllers was more efficient than without, thus keeping the pH difference between both sides of the membrane constant, which is a driving force for the uphill transport. Furthermore, the effect of pH of the acidic side on the uphill transport was investigated and the mechanism of the pH-controlled uphill transport is discussed.     

Japanese educational and learning systems in the post-industrial society: A value chain of educational and learning systems

Japan's educational system has some major problems. The most important among these concerns is the basic concept of the educational process and the goal of education. The old concept of public educational systems has become outdated in today's Japanese society, although this concept had supported social and spiritual faith, economic success and selfless devotion to one's country for more than 100 years. Now, Japanese people need a new concept of the educational process and the goal of education for the twenty-first century. The paper proposes a value chain of educational and learning systems aimed at building a network consisting of multiple fields for fostering future human resources.     

Evidence that activation of Src family kinase is not required for fertilization-associated [Ca2+]i oscillations in mouse eggs

Recent evidence in marine invertebrate, frog, and zebrafish eggs suggests the involvement of a Src family kinase (SFK) in fertilization-induced Ca2+ release. In the present study, we have investigated whether activation of an SFK is required for initiation of intracellular Ca2+ ([Ca2+]i) oscillations in mouse fertilization. We detected a Hck-like protein and tyrosine-phosphorylated proteins in soluble and insoluble sperm fractions, respectively. However, the presence of these proteins did not correspond to the active fractions of porcine sperm extracts (pSE). Moreover, [Ca2+]i oscillations induced by pSE in mouse eggs were unaltered by pre-incubation of pSE with specific SFK inhibitors such as 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazol[3,4-d]-pyrimidine (PP2) or lavendustin A, despite the fact that the inhibitors were shown to be active both in vivo and in vitro. Another SFK inhibitor, peptide A, blocked oscillations when incubated with pSE prior to injection into eggs, but this inhibition required more than ten times the concentration reportedly required to inhibit SFK activity. In addition, pre-injection or pre-incubation of eggs with these inhibitors did not affect the ability of pSE to trigger [Ca2+]i oscillations in mouse eggs. Microinjection of a recombinant c-Src protein or mRNAs encoding constitutively active Src proteins did not induce [Ca2+]i release. Finally, when sperm and eggs, both of which were pre-treated with PP2, were fertilized, [Ca2+]i oscillations occurred normally. We can therefore conclude that activation of an SFK is neither necessary nor sufficient for triggering fertilization-induced [Ca2+]i oscillations.     

Giant room-temperature magnetoresistance in single-crystal Fe/MgO/Fe magnetic tunnel junctions

The tunnel magnetoresistance (TMR) effect in magnetic tunnel junctions (MTJs) is the key to developing magnetoresistive random-access-memory (MRAM), magnetic sensors and novel programmable logic devices. Conventional MTJs with an amorphous aluminium oxide tunnel barrier, which have been extensively studied for device applications, exhibit a magnetoresistance ratio up to 70% at room temperature. This low magnetoresistance seriously limits the feasibility of spintronics devices. Here, we report a giant MR ratio up to 180% at room temperature in single-crystal Fe/MgO/Fe MTJs. The origin of this enormous TMR effect is coherent spin-polarized tunnelling, where the symmetry of electron wave functions plays an important role. Moreover, we observed that their tunnel magnetoresistance oscillates as a function of tunnel barrier thickness, indicating that coherency of wave functions is conserved across the tunnel barrier. The coherent TMR effect is a key to making spintronic devices with novel quantum-mechanical functions, and to developing gigabit-scale MRAM.     

Cooking up an interactive olfactory game display.

It's long been possible to give users outside an actual environment that environment's visual and auditory information and thus contribute to establishing presence. However, we've yet to establish much presence when users require olfactory information - such as in environments focused on foods, flowers, perfumes, or, in some cases, more offensive smells. Recently, several VR researchers have become interested in olfaction and olfactory displays that present smells in virtual environments (VEs). In this article, we describe our interactive olfactory display. One of our development goals is to confirm the assumption that users' interactions with the system increases presence. Thus, we used our interactive olfactory display to develop a cooking game in collaboration with electronic engineers and artists.     

Software System for Neuron Classification Based on Simple Parameters

A software system was produced to digitize neuronal images and to extract a set of simple parameters from the digitized images. The system was designed to classify a large number of neurons based on simple and automatically extracted features. The system was used to characterize catfish retinal ganglion cells. Several examples of parameter extraction and classification are presented.     

Capacitance-voltage characteristics of Semiconductor-Insulator-Semiconductor (SIS) structure

The ideal low- and high-frequency capacitance-voltage curves of a semiconductor(2)-insulator-semiconductor(1) (SIS) structure were first calculated with the insulator thickness, conductivity types and doping concentrations in semiconductor(1) and semiconductor(2) as parameters. The effects of fixed oxide charge and interface trap charge on the low and high frequency capacitance-voltage curves were also calculated. It was found that the fixed oxide sheet charge density with its centroid and the order estimation of the interface trap charge density with its effective type in addition to the insulator thickness, conductivity types and doping concentrations in semiconductor(1) and semiconductor(2) could be estimated from measured low and high frequency capacitance-voltage curves of an SIS structure.     

Millimeter-Wave Radiometer for the Measurement of Stratospheric ClO Using a Superconductive (SIS) Receiver Installed in the Southern Hemisphere

A new sensitive radiometer for atmospheric minor constituents has been developed by Nagoya University and the National Institute for Environmental Studies (NIES). The instrument equipped low-noise superconducting (SIS) mixer with a system noise temperature of ≈200K (SSB) at 204GHz. The SIS mixer is operated in a single side band mode with a side band ratio grater than 10dB by adjusting the two tunable backshorts of the mixer. Since any additional SSB filters are not necessary, we can simplify the quasi-optical system and reduce the standing waves which makes difficult to achieve a flat spectral baseline. We installed the observation system at Las Campanas Observatory, Chile in December 1999, and started the test observation from 2000 October. We have detected significant ClO emission at 204.3 GHz with an actual observation time of only 4 hours. This is the first detection of the ClO at 40km altitude from the mid-latitude region of the southern hemisphere. The Nagoya-NIES system has enabled continuous monitoring of ClO in the stratosphere with a time resolution of 4 or 5 hours.     

Analysis of plasticizers in cap-sealing resins for bottled foods

A survey of plasticizers in cap-sealing resins for bottled foods has been undertaken. During 1997-1999 di-(2- ethylhexyl)phthalate (DEHP) was found in seven out of 21 samples on the Japanese domestic market and in 10 out of 61 imported samples as well as a further two samples which contained di-(2-ethylhexyl)adipate (DEHA). In the period 1993-1999, of the other plasticizers diacetyl lauroyl glycerol (DALG) was only detected in domestic samples whereas diisodecyl phthalate (DIDP) and diisononyl phthalate (DINP) were only in imported samples. It was observed overall that DEHP and DEHA were restricted to use in cap-sealing resins for bottled foods. Whilst phthalates, DEHA or DALG were detected in samples in 1993 and 1995, the investigation in 1997-1999 showed fewer samples in which these plasticizers were found.