PCR-based identification of pathogenic Candida species using primer mixes specific to Candida DNA topoisomerase II genes

For rapid identification of Candida to the species level, degenerated primers and specific primers based on the genomic sequences of DNA topoisomerase II of C. albicans, C. dubliniensis, C. tropicalis (genotypes I and II), C. parapsilosis (genotypes I and II), C. krusei, C. kefyr, C. guilliermondii, C. glabrata, C. lusitaniae and Y. lipolytica were designed and their specificities tested in PCR-based identifications. Each of the specific primers selectively and exclusively amplified its own DNA fragment, not only from the corresponding genomic DNA of the Candida sp. but also from DNA mixtures containing other DNAs from several fungal species. For a simpler PCR-based identification, the specific primers were divided into three groups (PsI, PsII and PsIII), each of which contained four specific primer pairs. PCR with the primer mixes yielded four different sizes of PCR product, corresponding to each Candida sp. in the sample DNA. To obtain higher sensitivity of PCR amplification, sample DNAs were preamplified by the degenerated primer pair (CDF28/CDR148), followed by the main amplification using the primer mixes. By including this nested PCR step, 40 fg yeast genomic DNA was detected in the sample. Furthermore, we applied this nested PCR to a clinical diagnosis, using splenic tissues from experimentally infected mice and several clinical materials from patients. In all cases, the nested PCR amplifications detected proper DNA fragments of Candida spp., which were also identified by the standard identification tests. These results suggest that nested PCR, using primer mixes of the Candida DNA topoisomerase II genes, is simple and feasible for the rapid detection/identification of Candida to species level in clinical materials.     

Dioxin concentrations in the edible parts of Japanese common squid and saury

We examined the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dioxin-like PCBs) in muscle and gut tissues from Japanese common squid and saury. These body parts are often eaten in Japan, so it is important to measure their dioxin concentrations and evaluate the risks to consumers. The toxic equivalent (TEQ) concentrations in the squid gut samples (1.0 to 14 pg-TEQ/g fresh weight, n=3) were 50-fold larger than those in the muscle tissues (0.020 to 0.22 pg-TEQ/g fresh weight, n = 3) taken from the same samples. By contrast, the TEQ concentrations in the saury gut samples (0.35 to 0.63 pg-TEQ/g fresh weight, n=3) were only 1.1- to 1.7-fold greater than those in the muscle tissues (0.33 to 0.37 pg-TEQ/g fresh weight, n= 3) from the same samples. The TEQ contents in the squid gut tissues ranged from 60 to 990 pg-TEQ/squid, accounting for about 95% of the total dioxin content of the edible parts of the samples. By contrast, the TEQ contents in the saury gut tissues ranged from 4.4 to 12 pg-TEQ/saury, accounting for less than 25% of the total dioxin content of the edible parts of the samples. These tissues showed comparable PCDD/PCDF-congener and dioxin-like PCB-isomer profiles in both species. The results indicate that squid gut tissues occasionally contain high levels of dioxins, and consumption of this foodstuff could potentially significantly increase the dietary intake of dioxins.     

Shock-consolidated TiO2 bulk with pure anatase phases fabricated by explosive compaction using underwater shockwave

Titanium dioxide (TiO₂) bulk with pure anatase phases was fabricated by an explosive compaction technique using an underwater shockwave. Dynamic shock pressure of 6 GPa was used to consolidate anatase TiO₂ powders. Its microstructural, crystalline structural and photocatalytic characteristics were observed and measured by various techniques, such as X-ray diffraction (XRD), scanning electron microscopy (SEM) and photocatalytic activity measurement system. It was confirmed that the relative density of anatase TiO₂ compact is about 96% (3.73 g/cm³) of the theoretical density (3.89 g/cm³) and a strong surface bonding between particles is formed by a shock energy. In X-ray diffraction analysis, high purity anatase phases, broadened peaks due to lattice defects and decreased crystallite size were found. For the photocatalytic activities, the anatase TiO₂ compact was quite excellent compared to the commercial sintered TiO₂ bulk.     

In vitro selection of proteins that undergo covalent labeling with small molecules by thiol-disulfide exchange by using ribosome display

There is a great deal of interest in proteins that can bind covalently to target molecules, as they allow unambiguous experiments by tight binding to molecules of interest. Here, we report the generation of proteins that undergo covalent labeling with small molecules through in vitro selection by using ribosome display. Selection was performed from a mutant library of the WW domain with a biotinylated peptide as its binding target, in which the biotin and the peptide are connected by a disulfide bond. After five rounds of selection, we identified mutants carrying a particular cysteine mutation. The binding target reacted specifically with the selected mutant, even in the presence of other proteins, and resulted in the generation of biotin- or peptide-labeled WW domains by thiol-disulfide exchange. When the mutant was fused to a protein of interest, the fusion protein was also labeled with biotin. Thus, the characteristics of the selected mutant should be suitable as a tag sequence that can be covalently labeled with small synthetic molecules. These results indicate that the rapid and efficient generation of such proteins is possible by ribosome display.     

Effect of surface properties of silica nanoparticles on their cytotoxicity and cellular distribution in murine macrophages

In this study, complexes composed of poly-l-tyrosine (pLT) and single-walled carbon nanotubes (SWCNTs) were produced and the dispersibility of the pLT/SWCNT complexes in water by measuring the ζ potential of the complexes and the turbidity of the solution were investigated. It is found that the absolute value of the ζ potential of the pLT/SWCNT complexes is as high as that of SWCNTs modified with double-stranded DNA (dsDNA) and that the complexes remain stably dispersed in the water at least for two weeks. Thermogravimetry analysis (TGA) and visualization of the surface structures of pLT/SWCNT complexes using an atomic force microscope (AFM) were also carried out.     

A Novel Gateway Selection Technique for Throughput Optimization in Configurable Wireless Mesh Networks

Wireless mesh networks (WMNs) have attracted much attention due to their low up-front cost, easy network deployment, stable topology, robustness, reliable coverage, and so forth. These advantages are suitable for the disaster recovery applications in disaster areas, where WMNs can be advantageously utilized to restore network collapse after the disaster. In this paper, based on a new network infrastructure for WMNs, to guarantee high network performance, we focus on the issue of throughput optimization to improve the performance for WMNs. Owing to selecting different mesh router (MR) as the gateway will lead to different network throughput capacity, we propose a novel gateway selection technique to rapidly select the optimal MR as the gateway, in order to maximize the network throughput. In addition, we take into account the traffic distribution for the MR to eliminate traffic congestion in our method. The performance of our proposed method is evaluated by both numerical and simulated analysis. The simulation results demonstrate that the gateway selection method is effective and efficient to optimize the throughput for WMNs.     

Improvement of Emissions and Burning Limits in Burner Combustion using an Injector on the Concept of Fuel-Water Internally Rapid Mixing

This study tried to apply biomass fuel to clean burner combustion under high load conditions by water addition. A newly developed injector mixes fuel rapidly with water inside of the injector with support of atomizing air. The mixture composed of three-fluid is injected as spray into a flame stabilizer of burner. Investigation of emission performance of this injector shows that the internally rapid mixing type of injector is hopeful technique to introduce water into burner combustion. Combustion by this injector emits exceedingly less particulate matters at high load. NOx emission is strongly dependent on water flow rate.     

Gain control in L-band EDFAs by monitoring backward travelingC-band ASE

A novel and simple technique for gain flatness control is reported for gain shifted, long wavelength band (L-band) erbium-doped fiber amplifiers (EDFAs). Utilization of the backward traveling amplified spontaneous emission (ASE) in the C-band is analyzed with respect to controlling the gain tilt observed in the L-band when the total input power of the EDFA is changed. It is shown that a gain flatness of 0.6 dB/30 nm can be achieved over a dynamic range greater than 10 dB by using the backward traveling ASE power in the C-band as a monitor to adjust the copropagating pump power of the EDFA. The proposed technique eliminates the need to extract the output signals from the monitored ASE signal, demonstrating the suitability and simplicity of the proposed technique for wavelength division multiplexed applications     

Uniformity of gallium doped zinc oxide thin film prepared by pulsed laser deposition

Recently, transparent conducting oxide thin films have attracted attention for the application to transparent conducting electrodes. In this work, we evaluated the uniformity of electrical, optical and structural properties for gallium doped zinc oxide thin films prepared on the 10 × 10 cm² silica glass substrate by pulsed laser deposition. The resistivity, carrier concentration, mobility, bonding state and atomic composition of the film were uniform along in-plane and depth direction over the 10 × 10 cm² area of the substrate. The film showed the average transmittance of 81-87%, resistivity of 1.4 × 10^− 3 Ω cm, carrier concentration of 9.7 × 10²⁰/cm³ and mobility of 5 cm²/Vs in spite of the amorphous X-ray diffraction pattern. The gradual thickness distribution was found, however, the potential for large-area and low temperature deposition of transparent conducting oxide thin film using pulsed laser deposition method was confirmed.