Oxygen toxicity induces apoptosis in neuronal cells

1. A high oxygen atmosphere induced apoptosis in cultured neuronal cells including PC12 cells and rat embryonic cortical, hippocampal, and basal forebrain neurons associated with DNA fragmentation and nuclear condensation. 2. The sensitivity of CNS neurons to a high-oxygen atmosphere was the following order; cortex > basal forebrain > hippocampus. 3. Cycloheximide and actinomycin-D inhibited the apoptosis, indicating that it depends on new macromolecular synthesis. In contrast, cultured postnatal CNS neurons were resistant to oxidative stress. 4. Neurotrophic factors such as nerve growth factor (NGF), fibroblast growth factor (FGF), and epidermal growth factor (EGF) blocked the apoptosis induced by a high-oxygen atmosphere.     

Structural analysis of nucleic acids by using fluorescence resonance energy transfer (FRET)

We examined changes in the extent of fluorescence resonance energy transfer (FRET) between two different fluorochromes attached to a single oligonucleotide in the presence or absence of target nucleic acids with a specific sequence and a higher-ordered structure. In our system, FRET was maximal when probes were free in solution and a decrease in FRET was evidence of successful hybridization. Incubation of the probe with a single-stranded complementary oligonucleotide reduced the FRET. While, a small change in FRET was also observed when the probe was incubated with an oligonucleotide in which the target site had been embedded in a stable hairpin structure. These results indicate that this spectrofluorometric method and FRET probes can be used to estimate the efficacy of hybridization between a probe and its target site within highly ordered structures. It should help us to estimate the suitability of designed functional molecules, such as antisense DNA and RNA and ribozymes, that target to specific sites.     

Memory array architecture and decoding scheme for 3 V only sectorerasable DINOR flash memory

A memory array architecture and row decoding scheme for a 3 V only DINOR (divided bit line NOR) flash memory has been designed. A new sector organization realizes one word line driver per two word lines, which is conformable to tight word line pitch. A hierarchical negative voltage switching row decoder and a compact source line driver have been developed for 1 K byte sector erase without increasing the chip size. A bit-by-bit programming control and a low threshold voltage detection circuit provide a high speed random access time at low V_cc and a narrow program threshold voltage distribution. A 4 Mb DINOR flash memory test device was fabricated from 0.5 μm, double-layer metal, triple polysilicon, triple well CMOS process. The cell measures 1.8×1.6 μm² and the chip measures 5.8×5.0 mm ². The divided bit line structure realizes a small NOR type memory cell     

Comparative responsiveness of isolated saphenous, femoral and external carotid arteries and veins of dogs to dihydroergotamine mesylate (DEM) (author's transl)

Changes in tension of spiral strips of saphenous, femoral and external carotid arteries and veins were measured isometrically. DEM stimulated 6 preparations in almost the same concentration ranges (pD2 values were 8.64 to 8.40). However, the dose-response curves for DEM indicted variations in responsiveness among the different arteries and veins. Compared with norepinephrine (NE) (1)( the intrinsic activity of DEM was 0.002 on saphenous arteries, 0.05 on saphenous veins, 0.03 on femoral arteries, 0.18 on femoral veins, 0.13 on external carotid arteries and 0.13 on external carotid jugular veins. Thus DEM contracted more potently the venous strips from the hind limbs. IN femoral and external carotid arteries, antagonism of serotonin by DEM or methysergide was investigated. DEM displaced the dose-contractile response curves for serotonin in a noncompetitive manner, and the antagonistic response of DEM to serotonin was about 8 times more effective in external carotid arteries (pD'2 value=6.96) than in femoral arteries (pD'2 value=6.05). Methysergide, unlike DEM, antagonized the response to serotonin in a competitive manner at low doses but in a noncompetitive manner in high doses, and was fairly equal in antagonizing response to serotonin in external carotid arteries (pA2 value=that the therapeutic value of DEM in the treatment of orthostatic hypotension is due to its selective anti-serotonin activity on the external carotid arteries.     

Redistribution of Protein Biological Activity in a Freeze-Dried Cake

Lactate dehydrogenase (LDH) solutions were freeze dried in order to investigate the redistribution of the remaining LDH activity in the dried cake. D-Mannitol and a polysaccharide (κ-carrageenan, ι-carrageenan, or dextran) were mixed and used as stabilizers in freeze drying. It was found that the remaining LDH activity was significantly redistributed in the freeze-dried cake, and the enzyme concentration was not uniform along the axis of the dried cake due to variations in the freeze-drying conditions and stabilizer concentration. In some cases, active enzymes were highly localized in a specific portion of the dried cake. This was mainly due to the redistribution of the protein content. Further freeze-drying tests employing bovine serum albumin (BSA) and the above-mentioned polysaccharides showed that proteins (BSA) tended to accumulate in the lower portions of the dried cake, forming concentration gradients. Therefore, the concentration gradient in the freeze-dried cake was mainly due to the freezing process. This redistribution of protein content contributed considerably to the apparent redistribution of enzyme activity. The results suggest that a process that can control the redistribution of protein content in a freeze-dried cake would further improve the biological activity of freeze-dried proteins.     

Thermal degradation and flammability of nanocomposites composed of silica cross-linked to poly(methyl methacrylate)

The effect of polymer cross-linkages on thermal degradation of silica/poly (methyl methacrylate) (PMMA) nanocomposites is investigated using a single novel nanoparticle. Nanosilica surface treated with KH570, an organic surface treatment capable of free-radical polymerisation, was used to cross-link PMMA via an in situ method. Scanning electron microscopy was used to characterise nanosilica before use, while X-ray diffraction confirmed silica was well dispersed in PMMA. Thermogravimetric analysis (TGA) results showed that thermal degradation of silica cross-linked nanocomposites was significantly stabilised compared to PMMA, with a 30% reduction in the peak mass loss rate. Kinetic studies revealed the degradation of nanocomposites in this work abide by first-order kinetics, with an increase in the degradation activation energy of approximately 100?kJ?mol^?1. This is nearly double the improvement compared to conventional PMMA-silica nanocomposites in literature, showing dramatic enhancements to thermal stability. Analysis of high-temperature residuals from TGA tests suggest that cross-linked silica have increased char yields when compared with both PMMA and traditional silica nanocomposites. Cone Calorimetry results showed the materials in this work have reduced heat release rates compared to PMMA and traditional silica-PMMA nanocomposites.     

Optimal frequency tracking method for phase‐shift PWM inverter

This paper proposes a new resonant frequency tracking control method for full‐bridge‐type high‐frequency inverters. Whereas the ordinary phase‐locked loop (PLL) based frequency control method uses a current sensor and a voltage sensor, the proposed technique can achieve the same purpose with a single current sensor. In high‐frequency power supply systems using a PLL, it is impossible to perform power control with an inverter. Therefore, an active converter must be used for power control, and the system grows larger. On the other hand, high‐frequency inverters using the proposed control system simultaneously enable power control and achieve the same resonant frequency tracking as a PLL, and thus high‐frequency power supply systems become extremely simple. This paper explains in detail the principle underlying the control method, and presents an example of a circuit to realize it. The theory is backed up by using a prototype high‐frequency power supply system which actually employs the proposed control system, thereby demonstrating its practical utility in industry. © 2012 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc.     

Production of dipeptidyl peptidase IV inhibitory peptides from defatted rice bran

The insulinotropic hormone glucagon-like peptide-1 is metabolised extremely rapidly by the ubiquitous enzyme dipeptidyl peptidase IV (DPP-IV). Therefore, human DPP-IV is a key regulator involved in the prevention and treatment of type 2 diabetes. To simplify the method of producing an inhibitory peptide against DPP-IV, we focused on rice bran (RB) as a source and subjected proteins from defatted RB to enzymatic proteolysis using 2 commercial enzymes. The RB peptides produced with Umamizyme G exhibited 10 times the inhibitory activity as those produced with Bioprase SP. The half-maximal inhibitory concentration (IC₅₀) value of the RB peptides was 2.3 ± 0.1 mg/ml. Leu-Pro and Ile-Pro were identified as the inhibitory peptides among the RB peptides produced with Umamizyme G. Ile-Pro was the strongest DPP-IV inhibitor among the 15 Xaa-Pro dipeptides and Pro-Ile tested. Ile-Pro competitively inhibited DPP-IV (K_i = 0.11 mM). Mass spectrometry indicated that the contents of Leu-Pro and Ile-Pro in the RB peptides were 2.91 ± 0.52 μg/mg.     

Self-healing bio-based furan polymers cross-linked with various bis-maleimides

A series of bio-based self-healing polymers was prepared by cross-linking a furan polymer, poly(2,5-furandimethylene succinate), with bis-maleimide compounds by means of Diels–Alder reactions. In addition to the amount of the bis-maleimide linker, the molecular structure of the bis-maleimide played a key role in determining the extent of the Diels–Alder reaction and the mechanical and healing properties of the polymers. Bis-maleimides with phenylene rings markedly enhanced the tensile strength of the network polymers but hindered healing. In contrast, bis-maleimides with a flexible molecular structure tended to improve the polymer elongation, affording polymers with excellent healing ability. The efficiencies of self-healing (=healing without any external stimulus) and healing with an assistance of solvent (CHCl₃) were over 70% and 80%, respectively, for the network polymer crosslinked by bis-maleimide with flexible long-alkyl segment. Thus, our results indicate that the choice of bis-maleimide linker is an important factor for designing furan-maleimide–based self-healing polymers.     

Fluorescent superlattices of gold nanoparticles: A new class of functional materials

Fluorescent three-dimensional (3-D) superlattices of dansyl glutathione protected gold nanoparticles, with potential applications in molecular detection, have been synthesized at an air/water interface by controlling the pH of the nanoparticle suspension. The number of fluorophores per nanoparticle was calculated to be ∼127. Morphologies of the superlattice crystals were examined using scanning electron microscopy (SEM). Most of the crystals observed were triangular in shape. High-resolution transmission electron microscopy (HRTEM) and small angle X-ray scattering (SAXS) were used to study the packing of nanoparticles in these crystals. Both these studies showed that the nanoparticles were arranged in a face-centered cubic (fcc) pattern with a particle-particle distance (center-center) of ∼10.5 nm. Evolution of the crystal morphologies with time was also examined. The fluorescence properties of these triangles were studied using confocal fluorescence imaging and confocal Raman mapping, which were in good agreement with the morphologies observed by SEM. The superlattice exhibits near-infrared (NIR) absorption in the range 1100–2500 nm. Easy synthesis of such functional nanoparticle-based solids makes it possible to use them in novel applications. We utilized the fluorescence of dansyl glutathione gold superlattice crystals for the selective detection of bovine serum albumin (BSA), the major protein constituent of blood plasma, based on the selective binding of the naphthalene ring of the dansyl moiety with site I of BSA. Electronic Supplementary Material  Supplementary material is available for this article at and is accessible for authorized users. This article is published with open access at Springerlink.com