Preparation of silk protein sericin as mitogenic factor for better mammalian cell culture

We previously reported that sericin small (sericin-S), with a molecular weight that ranges from 5 to 100 kDa, is a cell culture supplement used to accelerate cell proliferation. In this study, a novel preparation method for sericin and several applications of sericin were examined. Sericin large, prepared under nonhydrolyzing conditions and ranging from 50 to 200 kDa, also accelerated cell proliferation, but its effects were inferior to those of sericin-S. Additional sericin preparations with various molecular weights that were differentially hydrolyzed were also tested but none of them was significantly superior to sericin-S, and neither were several recombinant sericin peptides. Sericin-S successfully accelerated the proliferation of hybridoma cells in various serum-free media, implying the mitogenic effect of sericin is independent from media. We also demonstrated that sericin-S successfully induced the proliferation of CTLL-2, an established T lymphocyte cell line, under IL-2 starvation conditions. These results indicate that sericin, particularly sericin-S, improves serum-free mammalian cell culture.     

Development and evaluation of qualitative detection methods for unapproved genetically modified rice (LLRice)

We developed a specific method to extract DNA from rice grain samples and modified the qualitative real-time PCR method provided by Bayer Co., Ltd. for reliable detection of the genetically modified (GM) rice variety, LLRice601, which has not undergone safety assessment for regulatory approval in Japan. Moreover, we conducted a data analysis to confirm the results obtained with real-time PCR. The yields of DNA extracted from powdered samples of rice grains were almost equal among 5 different varieties of rice, and there was no significant difference in the yield over three days. Reliable results were obtained using 50 ng of the extracted DNA as the template for real-time PCR. To examine the adequacy of the methods, we organized an interlaboratory study with the participation of 2 laboratories, in which 80 test samples were analyzed in a blinded manner. The statistical analysis revealed no significant difference in the Ct value for the endogenous gene of the DNA samples and for the targeted DNA sequence of 0.1% samples. The limit of detection of the method was approximately 0.1%. Analysis of the fluorescence intensity of the PCR-amplified product of the construct-specific DNA sequence suggested that it may be reasonable to judge a sample as positive when a Ct value of less than 40 is obtained.     

Effects of deletion of different PP2C protein phosphatase genes on stress responses in Saccharomyces cerevisiae

A key mechanism of signal transduction in eukaryotes is reversible protein phosphorylation, mediated through protein kinases and protein phosphatases (PPases). Modulation of signal transduction by this means regulates many biological processes. Saccharomyces cerevisiae has 40 PPases, including seven protein phosphatase 2C (PP2C PPase) genes (PTC1–PTC7). However, their precise functions remain poorly understood. To elucidate their cellular functions and to identify those that are redundant, we constructed 127 strains with deletions of all possible combinations of the seven PP2C PPase genes. All 127 disruptants were viable under nutrient‐rich conditions, demonstrating that none of the combinations induced synthetic lethality under these conditions. However, several combinations exhibited novel phenotypes, e.g. the Δptc5Δptc7 double disruptant and the Δptc2Δptc3Δptc5Δptc7 quadruple disruptant exhibited low (13°C) and high (37°C) temperature‐sensitive growth, respectively. Interestingly, the septuple disruptant Δptc1Δptc2Δptc3Δptc4Δptc5Δptc6Δptc7 showed an essentially normal growth phenotype at 37°C. The Δptc2Δptc3Δptc5Δptc7 quadruple disruptant was sensitive to LiCl (0.4 m ). Two double disruptants, Δptc1Δptc2 and Δptc1Δptc4, displayed slow growth and Δptc1Δptc2Δptc4 could not grow on medium containing 1.5 m NaCl. The Δptc1Δptc6 double disruptant showed increased sensitivity to caffeine, congo red and calcofluor white compared to each single deletion. Our observations indicate that S. cerevisiae PP2C PPases have a shared and important role in responses to environmental stresses. These disruptants also provide a means for exploring the molecular mechanisms of redundant PTC gene functions under defined conditions. Copyright © 2014 John Wiley & Sons, Ltd.     

Enclathration of hydrogen by organic-compound clathrate hydrates

The properties of hydrogen enclathration by cyclic ethers and acetone clathrate hydrates were investigated by powder X-ray diffraction, Raman spectroscopic analysis and volumetric analysis. Powder X-ray diffraction profiles indicate that the hydrates are structure-II hydrates. The variation in lattice constant by hydrogen occupation was investigated. This result indicates that inclusion of H₂ atom within empty small cage changes size of host cages depending on type of guest molecule. Raman results show that the samples formed binary clathrate hydrate of hydrogen and each organic compound. The amount of encaged H₂ was found to be comparable to that of H₂–THF binary hydrate. The trend of the changes for lattice constants is not related to the amount of encaged H₂. These results suggest that the organic compounds investigated in this study can be used as alternatives to THF for H₂ enclathration.     

Initial stage of carbon nanotube formation process by surface decomposition of SiC: STM and NEXAFS study

Formation process of carbon nanocaps, which are formed at the beginning of carbon nanotube (CNT) growth by surface decomposition of SiC, was investigated by scanning tunneling microscopy (STM) and in situ near-edge X-ray absorption fine structure (NEXAFS) spectroscopy. As Si atoms were desorbed, carbon nanoparticles 1-2 nm in diameter were accumulated on SiC(000-1) surfaces. At around 1200 °C, these were coalesced and crystallized to carbon nanocaps. In addition, just before the crystallization, majority of C-C bonds were directed nearly parallel to the surface. Based on these experimental results, we proposed a model for carbon nanocap formation, which plays an important role to determine the CNTs.     

Synthesis of epoxy resins from alcohol‐liquefied wood and the mechanical properties of the cured resins

New wood‐based epoxy resins were synthesized from alcohol‐liquefied wood. Wood was first liquefied by the reaction with polyethylene glycol and glycerin. The alcohol‐liquefied wood with plenty of hydroxyl groups were precursors for synthesizing the wood‐based epoxy resins. Namely, the alcoholic OH groups of the liquefied wood reacted with epichlorohydrin under alkali condition with a phase transfer catalyst, so that the epoxy groups were put in the liquefied wood. The wood‐based epoxy resins and the alcohol‐based epoxy resins as reference materials were cured with polyamide amine. The glass transition temperature (Tg), the tensile strength, and the modulus of elasticity of the wood‐based epoxy resin were higher than those of the alcohol‐based epoxy resin. Also, the shear adhesive strength of the wood‐based epoxy resin to steel plates was higher than those of the alcohol‐based epoxy resins, which was equivalent to the level of petroleum‐based bisphenol‐A type epoxy resins. The higher Tg of the wood‐based epoxy resin than that of the alcohol‐based epoxy resin is one of the evidences that the wood‐derived molecules were chemically incorporated into the network structures. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Cu‐rich CuInSe2 solar cells with a Cu‐poor surface

We present the effects of an In‐Se surface treatment of Cu‐rich ([Cu]/[In] > 1) CuInSe₂ absorbers in order to utilize the superior transport properties of Cu‐rich absorbers for high‐efficiency solar cells. A Cu‐poor surface was successfully formed on a Cu‐rich absorber by co‐evaporation of indium and selenium under appropriate conditions. The aim is to suppress the interface recombination, which is generally observed at the interface between CdS and Cu‐rich CuInSe₂. A surface‐treated device achieved an efficiency of 13.1%, improved from 9.5% obtained with an untreated Cu‐rich device. The open‐circuit voltage of In–Se treated devices reach the same level as devices made from Cu‐poor absorbers grown by a three‐stage process. By comparing the results of the experiments and simulations, it is shown that the role of the Cu‐poor surface on Cu‐rich absorbers is to prevent interface recombination, mainly due to a lower doping level of the Cu‐poor surface layer than in the Cu‐rich CuInSe₂. Thus, it becomes possible to uncouple the interface from the bulk properties in Cu‐rich solar cells. Copyright © 2014 John Wiley & Sons, Ltd.     

A novel catalyst fabricated from Al–Cu–Fe quasicrystal for steam reforming of methanol

The optimum preparation condition of Al–Cu–Fe quasicrystalline (QC) catalyst with excellent catalytic performance for steam reforming of methanol (SRM) has been investigated. The QC alloy is superior to the other crystalline Al–Cu–(Fe) alloys (i.e., beta and theta phase) as a catalyst material because of the brittle nature of QC. The wet milling process (in ethanol) for the QC powders is much better than the dry milling process to obtain fine particles with high surface area. The QC powder prepared by the wet process followed by leaching in Na₂CO₃ aq. at 323 K exhibited the highest catalytic performance (activity and stability) in the present study. From these findings, it is clear that the QC catalyst with the excellent catalytic performance could be obtained by controlling the initial grain size of the QC powder and the leaching temperature.