Isolation and structural analysis of efficient autonomously replicating sequences (ARSs) of the yeast Candida utilis

The industrially important yeast Candida utilis is widely used in production of food and medical materials, but its host-vector system has not been well developed. We screened for compact and efficient ARSs to construct practically useful vectors. The C. utilis strain AHU3053 was found to be efficiently transformed by the conventional lithium acetate method and was used as the host. The C. utilis IAM4264 genomic library was constructed by inserting the partial Sau3AI digests in pRI51, which has a kanMX gene expressible in C. utilis. By examining 98 C. utilis G418-resistant transformants, five plasmids had the highest ARS activity. By trimming of the inserts, the 1490 and 552 bp fragments with transformation activity of over 10(3)/microg DNA were obtained from ARS3 and ARS4, respectively. Although several sequences identical to S. cerevisiae ARS consensus sequences (ACSs) were found in ARS3 and ARS4, our deletion analysis indicated that these were not essential for the activity. Because the minimal functional ARS fragment was also several-fold larger than that of S. cerevisiae, the C. utilis ARSs have some unique characteristics resembling the Sz. pombe ARSs. These ARSs were functional in other C. utilis strains tested and useful for constructing practical vectors.     

Viewing angle performance of TN-LCD with hybrid aligned nematic film

We developed a novel hybrid aligned nematic film that improves viewing angle performance of twisted nematic liquid crystal display (TN-LCD). The viewing angle performance of the normally white (NW) TN-LCD with the compensation film was evaluated by both computer simulation and experiment. The compensated NW-TN-LCD exhibits wide viewing angle performance including good gray scale stability and low color shift.     

Effect of heating orientation on oscillatory thermocapillary flow in liquid bridges

Oscillatory thermocapillary flow in liquid bridges of high Prandtl number fluids is investigated experimentally as well as numerically. Heat loss from the free surface is shown to have significant influence on the onset of oscillations. In the present work the surrounding air flow pattern is changed by heating the liquid bridge from the top or from the bottom. The results show that the local free surface heat transfer rate, especially near the hot wall, has strong effects on the oscillations.     

Design and synthesis of potent antitumor 5,4'-diaminoflavone derivatives based on metabolic considerations

Recently, we reported that 5,4'-diaminoflavone (1) exhibits potent and specific growth-inhibitory activity against the estrogen receptor (ER)-positive human breast cancer cell line MCF-7. However, when compound 1 was incubated with S-9 mix, its metabolites were observed. Moreover, addition of S-9 mix to the medium caused the drastic decrease in activity of compound 1. Since the 6-, 8-, and 3'-positions were considered to be metabolized oxidatively in vivo from MO calculations, a series of 5,4'-diaminoflavone derivatives substituted at such putative metabolic positions with various functional groups were synthesized aiming at the metabolically stable derivatives. Among them, 5,4'-diamino-6,8,3'-trifluoroflavone (14d) exhibited strong growth-inhibitory activity against MCF-7 cells even in the presence of S-9 mix. Moreover, orally administered compound 14d completely suppressed the growth of MCF-7 inoculated into nude mice, and the effect was more potent than that of compound 1. In addition to ER-positive breast cancer cells, compound 14d exhibited growth-inhibitory activity against a panel of human cancer cell lines including a part of ER-negative breast, endometrial, ovarian, and liver cancers. From these results, fluorine introduction to the putative metabolic positions of compound 1 was elucidated to be effective in the enhancement of the in vivo antitumor activity, probably due to the block of the metabolic deactivation.     

Application of an electron backscatter diffraction pattern to Cu damascene-fabricated interconnections filled by a high-pressure anneal process

The microstructure of Cu interconnections fabricated by high-pressure annealing was evaluated using a field emission scanning electron microscope/electron backscatter diffraction pattern (FE-SEM/EBSP) technique, and the results are compared with as-deposited and normally annealed Cu films. The results show some grains extending from the bulk field to the via regions in the case of the high-pressure annealed Cu films. The existence of via holes was also observed, in which all grains were (111) oriented. This indicates that the high-pressure annealing process enables the Cu that in-fills the via holes to develop into favorable microstructures, i.e., single-crystal and with (111) orientation.     

Proposal of utilization of nuclear spent fuels for gamma cells

Large amounts of nuclear spent fuel are generated in nuclear power plants every year and stored in fuel storage facilities for 20–30 years until reprocessing. However, the spent fuel still has residual energies, such as high-temperature heat energy and high-intensity gamma radioactivity. We have examined the characteristics of solar cells exposed to gamma radiation for the development of gamma cells utilizing nuclear spent fuel. We used a highly intense ⁶⁰Co gamma source as a suitable substitute for spent fuel due to safety concerns and convenience. Two representative types of solar cells, amorphous and crystalline cells, were examined and the current and voltage generated by each type were measured. In general, solar cells are largely insensitive to gamma radiation because the radiation passes through solar cells without imparting all of its energy. In order to enhance the sensitivity to radiation, the solar cells were coupled to CsI(Tl), NaI(Tl) and plastic scintillators. We confirmed the following characteristics: (1) amorphous solar cells coupled to a CsI(Tl) scintillator are able to generate a large amount of electric power, compared to crystal-type solar cells, (2) amorphous cells exhibit a good linear response to high-intensity gamma radiation and generate electric power almost in proportion to the volume of the scintillator used, (3) the generated electric power is independent of the incident angle of the gamma rays and the amount of power is determined only by the volume of the scintillator used. The electric power generated by a single solar cell is very small, but a large amount of electric power can be obtained by arranging many solar cells in stacks and combining their induced current or voltage and by operating the cells all day, as they are not affected by weather conditions. We concluded that gamma cells utilizing the gamma radiation of nuclear spent fuel can be expected to be useful for electric power generation in the near future.     

Rapid and highly sensitive enzyme immunoassay for quantitative determination of tetrodotoxin

A monoclonal antibody against tetrodotoxin (TTX) was obtained from Balb/c mice immunized with TTX-bovine serum albumin (BSA) conjugate. The monoclonal antibody was highly specific for TTX and had no cross-reaction to tetrodonic acid, which is a TTX derivative, or gonyautoxins, although a minor cross-reaction to anhydro-tetrodotoxin was observed. The monoclonal antibody neutralized the lethal activity of TTX. By using the monoclonal antibody, a rapid and highly sensitive competitive enzyme immunoassay (EIA) for quantitative analysis of TTX was developed. By the competitive EIA system, TTX can be determined quantitatively in about 30 min (90 min are required if the time for preparation of the solid-phase antigen was included), and the working range for quantitative analysis of TTX was 2-100 ng/ml. In recovery tests and examinations of TTX samples, results of the mouse bioassay and EIA analyses correlated well (r = 0.987). Moreover, it was demonstrated that low concentrations of TTX, which could not be detected by the mouse bioassay, could be determined quantitatively by the competitive EIA.