Production of Lacto-N-biose I Using Crude Extracts of Bifidobacterial Cells

Lacto-N-biose I (LNB) is supposed to represent the bifidus factor in human milk oligosaccharides, and can be practically produced from sucrose and GlcNAc using four bifidobacterial enzymes, 1,3-β-galactosyl-N-acetylhexosamine phosphorylase, sucrose phosphorylase, UDP-glucose-hexose 1-phosphate uridylyltransferase, and UDP-glucose 4-epimerase, recombinantly produced by Escherichia coli. Here the production of LNB by the same enzymatic method without using genetically modified enzymes to consider the use of LNB for a food ingredient was reported. All four enzymes were produced as the intracellular enzymes of Bifidobacterium strains. The mixture of the crude extracts contained all four enzymes, with other enzymes interfering with the LNB production, namely, phosphoglucomutase, fructose 6-phosphate phosphoketolase, and glycogen phosphorylase. The first two interfering enzymes were selectively inactivated by heat treatment at 47 °C for 1 h in the presence of pancreatin, and glycogen phosphorylase was disabled by hydrolyzing its possible acceptor molecules using glucoamylase. Finally, 91 % of GlcNAc was converted into LNB in the 100-mL reaction mixture containing 300 mM GlcNAc.     

An Antisense Oligonucleotide against a Splicing Enhancer Sequence within Exon 1 of the MSTN Gene Inhibits Pre-mRNA Maturation to Act as a Novel Myostatin Inhibitor

Antisense oligonucleotides (ASOs) are agents that modulate gene function. ASO-mediated out-of-frame exon skipping has been employed to suppress gene function. Myostatin, encoded by the MSTN gene, is a potent negative regulator of skeletal muscle growth. ASOs that induce skipping of out-of-frame exon 2 of the MSTN gene have been studied for their use in increasing muscle mass. However, no ASOs are currently available for clinical use. We hypothesized that ASOs against the splicing enhancer sequence within exon 1 of the MSTN gene would inhibit maturation of pre-mRNA, thereby suppressing gene function. To explore this hypothesis, ASOs against sequences of exon 1 of the MSTN gene were screened for their ability to reduce mature MSTN mRNA levels. One screened ASO, named KMM001, decreased MSTN mRNA levels in a dose-dependent manner and reciprocally increased MSTN pre-mRNA levels. Accordingly, KMM001 decreased myostatin protein levels. KMM001 inhibited SMAD-mediated myostatin signaling in rhabdomyosarcoma cells. Remarkably, it did not decrease GDF11 mRNA levels, indicating myostatin-specific inhibition. As expected, KMM001 enhanced the proliferation of human myoblasts. We conclude that KMM001 is a novel myostatin inhibitor that inhibits pre-mRNA maturation. KMM001 has great promise for clinical applications and should be examined for its ability to treat various muscle-wasting conditions.     

High-rate Decoupled Water Electrolysis System Integrated with α-MoO3 as a Redox Mediator with Fast Anhydrous Proton Kinetics

Hydrogen is a promising alternative to fossil fuels that can reduce greenhouse gas emissions. Decoupled water electrolysis system using a reversible proton storage redox mediator, where the oxygen evolution reaction and hydrogen evolution reaction are separated in time and space, is an effective approach to producing hydrogen gas with high purity, high flexibility, and low cost. To realize fast hydrogen production in such a system, a redox mediator capable of releasing protons rapidly is required. Herein, α-MoO₃, with an ultrafast proton transfer property that can be explained by a dense hydrogen bond network in the lattice oxygen arrays of HxMoO₃, is examined as a high-rate redox mediator for fast hydrogen production in acidic electrolytes. The α-MoO₃ redox mediator shows both a large capacity of 204 mAh g⁻¹ and fast hydrogen production at a current rate of 10 A cm⁻²(≈153 A g⁻¹), outperforming most of the previously reported solid-state redox mediators.     

P-MOSFET's with ultra-shallow solid-phase-diffused drain structureproduced by diffusion from BSG gate-sidewall

A p-MOSFET structure with solid-phase diffused drain (SPDD) is proposed for future 0.1-μm and sub-0.1-μm devices. Highly doped ultrashallow p⁺ source and drain junctions have been obtained by solid-phase diffusion from a highly doped borosilicate glass (BSG) sidewall. The resulting shallow, high-concentration drain profile significantly improves short channel effects without increasing parasitic resistance. At the same time, an in situ highly-boron-doped LPCVD polysilicon gate is introduced to prevent the transconductance degradation which arises in ultrasmall p-MOSFETs with lower process temperature as a result of depletion formation in the p⁺-polysilicon gate. Excellent electrical characteristics and good hot-carrier reliability are achieved     

Scanning and transmission electron microscopic studies on the upper and lower surfaces of the frog skin epidermal cells.

We examined the fine structure of the upper and lower surfaces of stratified squamous epithelial cells in the skin of frogs (Hyla japonica). SEM revealed the upper surface of superficial cells covered with ramified microridges (type 3). The width of the microridges was 0.20-0.24 microns. Microridges found at the cell boundary were about 0.30 microns in width and a narrow furrow was seen between the two cells. The numerous oval disk-like structures (0.23 x 0.32 microns in diameter) covered the lower surface of these superficial cells. The upper surface of cells in the 2nd layer was covered with baculiform or ramified microridges (type 2 or 3). On the cell boundary, two linear microridges (0.23-0.27 microns in width) were parallelly arranged. The width of the microridges covering the upper cell surface was 0.09-0.10 microns. Microvilli-like processes with a height of 0.32-0.37 microns were interspersed among the microridges. Their tip formed an oval plane (0.23 x 0.31 microns), which corresponded to the size of the disk-like structures on the lower surface of the superficial cells. Desmosomes were observed on the tip by TEM. These findings show that the disk-like structures on the lower surface of the superficial cells are the sites of binding with the microvilli-like processes on the upper surface of the 2nd layer cells. The disk-like structures observed in the present study seem to be equivalent to the binding site on the upper surface of the surface cell layer of mammalian stratified squamous epithelium.     

Fibre transmission distance determined by eye opening degradation due to selfphase modulation and group-velocity dispersion

Numerical analysis of IM signal propagation in an optical fibre is carried out taking selfphase modulation and group-velocity dispersion into account. The transmission distance yielding a prescribed eye opening penalty, in the normal dispersion region, is shown to be inversely proportional to the square root of the signal power.<>     

DNA Computing Boosted by a Cationic Copolymer

The huge information storage capability of DNA and its ability to self‐assemble can be harnessed to enable massively parallel computing in a small space. DNA‐based logic gates are designed that rely on DNA strand displacement reactions; however, computation is slow due to time‐consuming DNA reassembly processes and prone to failure as DNA is susceptible to degradation by nucleases and under certain solution conditions. Here, it is shown that the presence of a cationic copolymer boosts the speed of DNA logic gate operations that involve multiple and parallel strand displacement reactions. Two kinds of DNA molecular operations, one based on a translator gate and one on a seesaw gate, are successfully enhanced by the copolymer without tuning of computing conditions or DNA sequences. The copolymer markedly reduces operation times from hours to minutes. Moreover, the copolymer enhances nuclease resistance.     

Fabrication and current-drive of SiGe/Si 'Micro-origami' epitaxial MEMS device on SOI substrate

A micromirror structure with SiGe/Si heteroepitaxial layer on a silicon-on-insulator (SOI) substrate using a 'Micro-origami' technique has been successfully fabricated. The micromirror is supported by two curved hinge structures. The device is driven by application of a current, and net angular displacements larger than 10/spl deg/ (static) and 30/spl deg/ (in resonance) were obtained. These values are comparable with or even larger than the reported values for other MEMS optical switches or beam scanning devices. The experimental results suggest that the movement is evoked by a thermal effect. The Micro-origami device has advantages of low operation voltage smaller than 2 V, and structural compatibility with the Si or SiGe LSIs.     

Annealing effects of a high-quality ZnTe substrate

The sharp photoluminescence (PL) and optical-reflection spectra in the bandedge region of the high-quality nondoped ZnTe substrate (100) were observed at 4.2 K. Free exciton, associated with lower and upper polaritons (EX^L and EX^U) at 2.382 eV and 2.381 eV, respectively, were clearly observed. This meant that this substrate was high quality. The intensity of a bound exciton peak (2.375 eV), which is caused by a Zn vacancy, of a neutral acceptor decreased with an increase of the Zn vapor pressures.