Detection of a tryptophan radical as an intermediate species in the reaction of horseradish peroxidase mutant (Phe-221 --> Trp) and hydrogen peroxide

The crucial reaction intermediate in the reaction of peroxidase with hydrogen peroxide (H2O2), compound I, contains a porphyrin pi-cation radical in horseradish peroxidase (HRP), which catalyzes oxidation of small organic and inorganic compounds, whereas cytochrome c peroxidase (CcP) has a radical center on the tryptophan residue (Trp-191) and oxidizes the redox partner, cytochrome c. To investigate the roles of the amino acid residue near the heme active center in discriminating the function of the peroxidases in these two enzymes, we prepared a CcP-like HRP mutant, F221W (Phe-221 --> Trp). Although the rapid spectral scanning and stopped-flow experiments confirmed that the F221W mutant reacts with H2O2 to form the porphyrin pi-cation radical at the same rate as for the wild-type enzyme, the characteristic spectral features of the porphyrin pi-cation radical disappeared rapidly, and were converted to the compound II-type spectrum. The EPR spectrum of the resultant species produced by reduction of the porphyrin pi-cation radical, however, was quite different from that of compound II in HRP, showing typical signals from a Trp radical as found for CcP. The sequential radical formation from the porphyrin ring to the Trp residue implies that the proximal Trp is a key residue in the process of the radical transfer from the porphyrin ring, which differentiates the function of peroxidases.     

A novel strategy of cell targeting based on tissue-specific expression of the ecotropic retrovirus receptor gene

Gene transfer into specific tissues or cell types is a key technique in the development of gene therapy. Modification of vector particles such that they selectively bind to the target cells has been attempted, but the limitation of this approach is the low transduction efficiency. Here, we show that a two-step gene transfer system can be used for efficient cell targeting. With this strategy, and using a high-titer adenoviral vector containing a tissue-specific promoter, we have engineered a system in which only target cells become susceptible to retrovirus-mediated transduction. In a model experiment, we constructed an adenoviral vector (Ad.AFPEcoRec) containing the ecotropic retrovirus receptor (EcoRec) gene under the control of the alpha-fetoprotein (AFP) promoter. A binding assay showed that after transduction with AD.AFPEcoRec, EcoRec molecules were efficiently expressed in AFP+HepG2 cells, but not in AFP-HeLa and AFP-HLE cells. The EcoRec-expressing HepG2 cells could be stably transduced with ecotropic retroviral vectors, whereas HeLa and HLE cells remained highly resistant to retrovirus-mediated gene transfer. The apparent titer on HepG2 cells was greater than 2 x 10(5) CFU/ml. Because various tissue-specific promoter/enhancer elements are available, the two-step system could be used as a general strategy for both ex vivo and in vivo targeted gene transfer.     

Effect of grain size on creep of Ti-53.4mol%Al intermetallics at 1100 K

Constant stress creep under compression stress, 100 to 316 MPa, at 1100 K was investigated on single-phase TiAl intermetallics. The material was ingot-cast, isothermally forged, and then annealed to produce stable equi-axed grain structures, whose average grain diameters were 25, 42 and 70m. Creep curves were very similar among the three specimens with different grain diameters and the creep rates at a given strain, as well as the minimum creep rates, depended little on grain size. Two regimes were observed on the stress dependence of the minimum creep rate. The stress exponent under high stresses was about 4.5, independent of grain size. Under stresses lower than about 150 MPa it became about 8.     

Preparation of oriented gold plate/cellulose nanofiber composite films by using TEMPO-oxidized cellulose nanofiber as a reducing agent

Microsystem Technologies - TEMPO-oxidized cellulose nanofiber (TOCN) has been drawing much attention as a useful material for preparation of transparent and stiff films or substrates. In this work,...     

A novel quarter‐wave retardation film for improving viewing angle properties in time‐sequential stereoscopic 3D‐LCDs

We have successfully developed a quarter‐wave retardation film (QWF) for wide viewing angle 3D liquid crystal displays (3D‐LCDs) that provides high luminance, low crosstalk, low color change, and low head‐tilt‐angle dependency. It was found that the out‐of‐plane retardation (Rth) of the QWF in the LCD needs to be close to 0 nm in order to improve the 3D display properties at an off‐axis position and that the in‐plane retardation (Re) needs to be adjusted from 120 to 130 nm to achieve low color change with head tilting. We adopted a coating process for making our QWF because of its potential for retardation control. 3D‐LCDs with this QWF whose Rth was nearly zero had high performance and allowed off‐axis other than on‐axis.     

Cognitive model of team cooperation in en-route air traffic control

Since controller teams are in charge of en-route air traffic control, team cooperation is a key issue for good control performance. We conducted ethnographic field observation at the Tokyo Area Control Center and then analyzed the obtained data to develop a cognitive model of team cooperation in en-route air traffic control. We segmented conversational records, behavioral records, and so on by control unit, and then clarified relations between the segments and identified expert knowledge and judgment behind them. Cognitive processes of controller teams were reconstructed based on a concept of distributed cognition. The analysis revealed that the mutual belief model is applicable to team cooperation processes, the role assignment of tasks within a controller team is implicit and that control plans are implemented smoothly once team situation awareness has been established. A cognitive model of controllers?? team cooperation has been constructed based on these findings.     

Effects of 2-Hz and 60-Hz electrical stimulation on the microstructure of beef

A comparative electron-microscope study was made of the structural changes brought about in bovine longissimus dorsi muscle by 2-Hz and 60-Hz electrical stimulation. Apart from the frequency difference, the two treatments were identical: 500V AC, 600 pulses (2Hz × 300s and 60 Hz × 10 s), and a time of application of about 40 min post mortem. The low-frequency current produced no detectable tissue disruption or irregularity in sarcomere length, whereas the 60-Hz treatment caused widespread supercontraction and the formation of contraction nodes; many sarcomeres in the internodal zones were stretched or fractured. The observations confirm and extend those of our earlier light-microscope study, and support the view that fiber disruption contributes significantly to the tenderizing effect of high-voltage electrical stimulation.     

Interdiffusion inα solid solutions of the Cu-Zn-Sn system

The interdiffusion coefficients in the f c c phase of Cu-Zn-Sn alloys, , have been determined at 1073 K. The concentration profiles indicate that the diffusion rate of tin is greater than that of zinc in the Cu-Zn-Sn alloy. The diffusion paths show the typical S-shaped curves. All of the four interdiffusion coefficients are positive and they are very sensitive to the solute concentration. The atomic mobilities of the three diffusing elements in Kirkendall planes increase in the order of Cu, Zn, Sn. The interaction energy of the Cu-Sn bond is much larger than that of the Zn-Sn bond. From the results of the present work it seems that the Onsager reciprocal relation holds in the a phase of the Cu-Zn-Sn system.     

Treatment of Oxidative Stress with Exosomes in Myocardial Ischemia

A thrombus in a coronary artery causes ischemia, which eventually leads to myocardial infarction (MI) if not removed. However, removal generates reactive oxygen species (ROS), which causes ischemia–reperfusion (I/R) injury that damages the tissue and exacerbates the resulting MI. The mechanism of I/R injury is currently extensively understood. However, supplementation of exogenous antioxidants is ineffective against oxidative stress (OS). Enhancing the ability of endogenous antioxidants may be a more effective way to treat OS, and exosomes may play a role as targeted carriers. Exosomes are nanosized vesicles wrapped in biofilms which contain various complex RNAs and proteins. They are important intermediate carriers of intercellular communication and material exchange. In recent years, diagnosis and treatment with exosomes in cardiovascular diseases have gained considerable attention. Herein, we review the new findings of exosomes in the regulation of OS in coronary heart disease, discuss the possibility of exosomes as carriers for the targeted regulation of endogenous ROS generation, and compare the advantages of exosome therapy with those of stem-cell therapy. Finally, we explore several miRNAs found in exosomes against OS.