A sequencing problem for a mixed-model assembly line in a JIT production system

In an assembly line of a just-in-time (JIT) production system, workers have the power and the responsibility to stop the line when they fail to complete their operations within their work zones. This paper deals with a sequencing problem for the mixed-model assembly conveyor line in the JIT production system. In some environment, the most important criterion is the line stoppage rather than the variation of production rates. The problem is to find an optimal sequence of units that minimizes the total line stoppage time. Lower and upper bounds of the total line stoppage time are derived and the branch-and-bound method is applied to the problem. A numerical example is given.     

Decreased membrane fluidity and unsaturated fatty acids in Niemann-Pick disease type C fibroblasts

We have investigated transformation with heterologous DNA as a method for insertional mutagenesis of Aspergillus fumigatus. Two methods, polyethylene glycol-mediated transformation of protoplasts and electroporation of germinating spores, were used to establish conditions leading to single-copy integration of transforming DNA at different genomic sites. We have assessed the effect of restriction enzyme-mediated integration (REMI) for both methods. Non-REMI protoplast transformation led to integration of multiple copies of transforming DNA in the majority of transformants. Results of REMI with protoplast transformation varied depending on the enzyme used. Low concentrations of several restriction enzymes stimulated transformation, but of ten enzymes investigated only REMI with XhoI and KpnI resulted in single-copy integration of transforming DNA for the majority of transformants. For protoplast transformation with XhoI- or KpnI-based REMI, 50% and 76% of insertions, respectively, were due to integrations at a genomic enzyme site corresponding to the enzyme used for REMI. Electroporation of spores without addition of restriction enzyme resulted in a high transformation efficiency, with up to 67% of transformants containing a single copy of transforming DNA. In contrast to protoplast transformation, electroporation of spores in the presence of a restriction enzyme did not improve transformation efficiency or lead to insertion at genomic restriction sites. Southern analysis indicated that for both protoplast transformation with REMI using KpnI or XhoI and for electroporation of spores without addition of restriction enzymes, transforming DNA inserted at different genomic sites in a high proportion of transformants.     

Titania NOx sensors for exhaust monitoring

Oxide semiconductors have been examined to develop NO_x sensors for exhaust monitoring. Titania doped with trivalent elements, such as Al³⁺, Sc³⁺, Ga³⁺ or In³⁺, has a good sensitivity and selectivity to NO between 450 and 550 °C, and shows rapid response. A sensor probe for monitoring exhaust NO_x has been fabricated. Many kinds of interference gases, such as C₃H₆, CO and SO₂, have been found to have only a slight influence on the sensor response to NO. The influence of O₂ and H₂O is also negligible, except for the cases of 0% H₂O and fuel-rich conditions. In accordance with these results, the sensor probe operates satisfactority in the exhaust gas of various combustion conditions without interference from the various kinds of gas species in the exhaust gases.     

Pseudo-binary system Bi2O3-TeO2 in air

The phase equilibria in the pseudo-binary system Bi₂O₃-TeO₂ at 600° 950° C in air were examined by solid-state reaction techniques and X-ray powder diffraction method. Four pseudo-binary compounds appeared, i.e., -Bi₂O₃ type solid solution having a compositional range of (1-x)Bi₂O₃·xTeO₂ wherex=0 0.4 a new compound Bi₆Te₂O₁₅ which has an orthorhombic cell of a=2.27(4) nm, b=1.06(1) nm and c = 0.539(8) nm, 2Bi₂O₃ · 3TeO₂, and an unidentified phase Bi₂O₃·2TeO₂. The formation of the phase Bi₆Te₂O₁₅, in which all the Te ions are hexavalent, was confirmed by the thermogravimetry and by the Mössbauer spectra. The liquidus curves for whole system were determined by DTA method.     

One-Step Generation of Multiple Gene-Edited Pigs by Electroporation of the CRISPR/Cas9 System into Zygotes to Reduce Xenoantigen Biosynthesis

Xenoantigens cause hyperacute rejection and limit the success of interspecific xenografts. Therefore, genes involved in xenoantigen biosynthesis, such as GGTA1, CMAH, and B4GALNT2, are key targets to improve the outcomes of xenotransplantation. In this study, we introduced a CRISPR/Cas9 system simultaneously targeting GGTA1, CMAH, and B4GALNT2 into in vitro-fertilized zygotes using electroporation for the one-step generation of multiple gene-edited pigs without xenoantigens. First, we optimized the combination of guide RNAs (gRNAs) targeting GGTA1 and CMAH with respect to gene editing efficiency in zygotes, and transferred electroporated embryos with the optimized gRNAs and Cas9 into recipient gilts. Next, we optimized the Cas9 protein concentration with respect to the gene editing efficiency when GGTA1, CMAH, and B4GALNT2 were targeted simultaneously, and generated gene-edited pigs using the optimized conditions. We achieved the one-step generation of GGTA1/CMAH double-edited pigs and GGTA1/CMAH/B4GALNT2 triple-edited pigs. Immunohistological analyses demonstrated the downregulation of xenoantigens; however, these multiple gene-edited pigs were genetic mosaics that failed to knock out some xenoantigens. Although mosaicism should be resolved, the electroporation technique could become a primary method for the one-step generation of multiple gene modifications in pigs aimed at improving pig-to-human xenotransplantation.     

Chemical Modification of Phage-Displayed Helix-Loop-Helix Peptides to Construct Kinase-Focused Libraries

Conformationally constrained peptides hold promise as molecular tools in chemical biology and as a new modality in drug discovery. The construction and screening of a target-focused library could be a promising approach for the generation of de novo ligands or inhibitors against target proteins. Here, we have prepared a protein kinase-focused library by chemically modifying helix-loop-helix (HLH) peptides displayed on phage and subsequently tethered to adenosine. The library was screened against aurora kinase A (AurA). The selected HLH peptide Bip - 3 retained the α-helical structure and bound to AurA with a K_D value of 13.7 μM. Bip - 3 and the adenosine-tethered peptide Bip - 3 - Adc provided IC₅₀ values of 103 μM and 7.7 μM, respectively, suggesting that Bip - 3 - Adc bivalently inhibited AurA. In addition, the selectivity of Bip - 3 - Adc to several protein kinases was tested, and was highest against AurA. These results demonstrate that chemical modification can enable the construction of a kinase-focused library of phage-displayed HLH peptides.     

Measurement of internal stresses in coatings using time resolved fluorescence

This paper describes a new nondestructive technique for detecting internal stresses in coatings using time resolved fluorescence. The measurement principle is based upon an experimental result that the decay time of fluorescence from poly(3-octylthiophene), P3OT, dispersed in uniaxially-stretched polymer films decreases with increasing tensile stress acting on the films. Internal stresses in the clear coat and in the base coat of a multilayer structure, which was composed of electrodeposited coat, surface coat, base coat, and clear coat, were estimated from the decay time of fluorescence from P3OT in both coats. The order of internal stresses in the clear coat and base coat of the multilayer system was 1 MPa. When the coatings were piled up, the internal stress decreased as the distance from a metal substrate was increased. It was found that moisture and temperature influenced internal stresses in the clear coat rather than in the base coat. Internal stress in the clear coat, which was one layer coated on a metal substrate, was measured by the traditional bimetallic method or by the time resolved fluorescence technique. Comparing both methods, it was concluded that the time resolved fluorescence technique gave reliable values for internal stresses in coatings.     

Adsorption properties of oxygen and methane on Ga-ZSM-5; the origin of the selectivity of NOx reduction using methane

The adsorption properties of oxygen and methane on Ga-ZSM-5 and Cu-ZSM-5 catalysts were examined by a TPD method to clarify the extraordinary selectivity of HC-SCR using methane on Ga-ZSM-5. It was observed that Ga-ZSM-5 did not adsorb oxygen but adsorbed methane dissociatively, while on Cu-ZSM-5 oxygen was dissociatively adsorbed and reacted with adsorbed ethylene.     

Characterization of Silicon Carbide–Silicon Nitride Composite Ultrafine Particles Synthesized Using a CO2 Laser by Silicon-29 Magic Angle Spinning NMR and ESR

The structure of silicon carbide–silicon nitride (SiC–Si₃N₄) composite particles synthesized using a CO₂ laser was studied by magic angle spinning nuclear magnetic resonance (MAS-NMR) and electron spin resonance (ESR). The structure around Si atoms changed by introducing N. C atoms around Si were substituted by N atoms, and N-rich configurations around Si atoms increased stepwise as the N content increased. The low N content composite particles consisted of mainly SiC phase containing dissolved N. N atoms were partly present in β-SiC microcrystal and partly in the grain boundary layer in the particle. N atoms were tetrahedrally surrounded by four Si atoms in β-SiC microcrystal and were trivalent state bonded to three Si atoms in the grain boundary layer. The high N content particles consisted of SiC, Si₃N₄, and amorphous phases, whose amount depended on N content.