Damage of yeast cells induced by pulsed light irradiation

DNA damage, such as formation of single strand breaks and pyrimidine dimers was induced in yeast cells after irradiation by pulsed light, which were essentially the same as observed with continuous ultraviolet (UV) light. The UV-induced DNA damage is slightly higher than seen with pulsed light. However, increased concentration of eluted protein and structural change in the irradiated yeast cells were observed only in the case of pulsed light. A difference in the inactivation effect between pulsed light and UV light was found and this suggested cell membrane damage induced by pulsed light irradiation. It is proposed that pulsed light can be used as an effective sterilizing method for the yeast Saccharomyces cerevisiae.     

Effect of direct ovarian injection of vascular endothelial growth factor gene fragments on follicular development in immature female rats

Vascular endothelial growth factor (VEGF) expression in granulosa cells is associated with the thecal vasculature growth during ovarian follicular development. We hypothesized that injection of VEGF gene fragments directly into the rat ovary would induce production of a large number of ovulatory follicles and that these follicles would ovulate. To test this hypothesis, we treated immature female rats with combinations of hormones and VEGF gene fragments. The animals were divided into two groups: one group received solution containing transfection reagents as a control (n = 5), while the other group received direct ovarian injection of VEGF gene fragments at 19 (n = 5), 21 (n = 5), 23 (n = 5), or 25 (n = 5) days after birth followed by i.p. administration of 20 IU equine chorionic gonadotropin (eCG) at the age of 26 days. Forty-eight hours after eCG injection, animals were given 20 IU human chorionic gonadotropin (hCG) i.p. and then the oocytes in both groups were counted. The maximum number of ovulated oocytes was obtained when the VEGF gene fragments were injected into the rat ovary at 21 days after birth. Histological examination revealed that the injection of VEGF gene fragments markedly increased the vascular density around the preovulatory follicles and also the number of these follicles. Our data provide the first reported evidence that most ovulatory follicles generated by injection of VEGF gene fragments are able to ovulate upon hCG treatment. These results demonstrate that injection of VEGF gene fragments directly into the ovary stimulates the development of antral follicles by inducing the formation of thecal vasculature in immature female rats.     

Effects of cubic BN addition and phase transformation on hardness of Al2O3-cubic BN composites

The Vickers hardness of dense Al₂O₃-cubic BN (cBN) composites prepared by spark plasma sintering under a moderate pressure of 100 MPa at 1200-1600 °C was investigated at indentation loads of 0.098-19.6 N. The BN grains in the Al₂O₃-BN composite prepared at 1300 °C showed no transformation from the cBN to hBN phase, and the hardness was 59 GPa at 0.098 N. The hardness of the Al₂O₃ matrix in the Al₂O₃-BN composites containing 10-30 vol% cBN prepared at 1300-1400 °C was around 25 GPa at 0.098 N, which was higher than monolithic Al₂O₃ bodies prepared at the same temperatures. The hardness of the Al₂O₃ matrix in the Al₂O₃-BN composites decreased with increasing sintering temperature. The increase in the hardness of the Al₂O₃ matrix may be due to the decrease in the size of Al₂O₃ grains in the Al₂O₃-BN composites owing to the addition of cBN particles and the decrease in sintering temperature. The Meyer exponents of the monolithic Al₂O₃ bodies and Al₂O₃-BN composites were 1.90-1.94 independent of cBN content.     

A new modeling approach for circuit‐field‐coupled time‐stepping electromagnetic analysis of saturated interior permanent magnet synchronous motors

This paper proposes a new modeling approach for circuit‐field‐coupled time‐stepping electromagnetic analysis of a saturated interior permanent magnet synchronous motor (IPMSM). To predict the drive performance quickly, the proposed approach consists of a dynamic simulator based on a new analytical model of the d‐ and q‐axis magnetization characteristics (λ‐i_d‐i_q‐θ). The model can take into account not only cross‐saturation but also the harmonics of the inductance distributions and EMF waveforms. The validity of the model is verified from suitable simulation results for the instantaneous current and torque waveforms of the IPMSM. The proposed analysis leads to a dramatic reduction in the computation time compared to circuit‐field‐coupled time‐stepping FEA, while maintaining analytical accuracy. The effectiveness of the proposed method is experimentally verified using a 10‐kW, 15,000‐rpm concentrated‐winding IPM motor. © 2010 Wiley Periodicals, Inc. Electr Eng Jpn, 174(1): 49–58, 2011; Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/eej.21024     

Analysis of enoyl-coenzyme A hydratase activity and its stereospecificity using high-performance liquid chromatography equipped with chiral separation column

Enoyl-coenzyme A (CoA) hydratase catalyzes the hydration of trans-2-enoyl-CoA to yield 3-hydroxyacyl-CoA during fatty acid degradation (β-oxidation). Although much research has focused on the stereospecificities of 2-enoyl-CoA hydratases, a direct quantification of the production of 3(R)- and 3(S)-hydroxyacyl-CoA has not yet been established. Therefore, we developed a method of concurrently quantifying 3(R)- and 3(S)-hydroxyacyl-CoA using high-performance liquid chromatography (HPLC) equipped with a chiral separation column. The optimized conditions for the separation of 3(R)-, 3(S)-hydroxyhexadecanoyl-CoA and trans-2-hexadecenoyl-CoA, were determined to be as follows: mobile phase of 35/65 (v/v) of 50 mM phosphate buffer (pH 5.0)/methanol; flow rate of 0.5 mL/min; detection at 260 nm; and column temperature of 25°C. This method was applied to subcellular fractions of rat liver; the results directly confirmed that 3(S)-hydroxyhexadecanoyl-CoA is the dominant product obtained from the heat-stable enoyl-CoA hydratase-catalyzed reaction of trans-2-hexadecenoyl-CoA. Finally, the stereospecificities of L-bifunctional protein (L-BP) and D-bifunctional protein (D-BP) were reinvestigated using this method, and it was confirmed that L- and D-BP yielded 3(S)- and 3(R)-hydroxyhexadecanoyl-CoA were yielded from trans-2-hexadecenoyl-CoA, respectively. 3(R)-Hydroxyacyl-CoA is a peroxisomal β-oxidation-specific intermediate. Therefore, this method is potentially useful not only studies regarding the stereochemistry of enoyl-CoA hydratase but also for the diagnosis of diseases caused by defects of peroxisomal enoyl-CoA hydratase.     

Detection of cytochrome P450 substrates by using a small-molecule droplet array on an NADH-immobilized solid surface

Seeing below the surface: A small-molecule droplet array platform on an NADH-immobilized solid surface and a biotinylated acetophenone derivative were developed to identify the substrate candidates for soluble P450 enzymes of interest. This methodology is thought to be easily applicable to other class I P450 systems, including those that use NADPH as cofactor.     

Melt-mixing by novel pitched-tip kneading disks in a co-rotating twin-screw extruder

Melt-mixing in twin-screw extruders is a key process in the development of polymer composites. Quantifying the mixing performance of kneading elements based on their internal physical processes is a challenging problem. We discuss melt-mixing by novel kneading elements called “pitched-tip kneading disk (ptKD)”. The disk-stagger angle and tip angle are the main geometric parameters of the ptKDs. We investigated four typical arrangements of the ptKDs, which are forward and backward disk-staggers combined with forward and backward tips. Numerical simulations under a certain feed rate and screw revolution speed were performed, and the mixing process was investigated using Lagrangian statistics. It was found that the four types had different mixing characteristics, and their mixing processes were explained by the coupling effect of drag flow with the disk staggering and pitched-tip and pressure flows, which are controlled by operational conditions. The use of a pitched-tip effectively controls the balance of the pressurization and mixing ability.