Takushi Togashi

Second-harmonic generation from a KBe(2) BO(3)F(2) crystal in the deep ultraviolet.

By use of KBe(2)BO(3)F(2) (KBBF) crystal with a size of 10 mmx10 mm x1.2 mm and a special prism-coupling technique (PCT), fourth-harmonic generation of Ti:sapphire laser systems from 200 to 179.4 nm has been achieved. Moreover, with a Ti:sapphire laser with a 50-fs pulse duration and a 1-kHz repetition rate, conversion efficiency as high as 13% from 400 to 200 nm without any surface-loss correction has also been obtained. The data show that with the PCT a KBBF crystal can produce deep-UV coherent light with measurable power output.

GENERATION OF 0.66-TW PULSES AT 1 KHZ BY A TI:SAPPHIRE LASER

A 1-kHz, 0.66-TW Ti:sapphire laser has been developed. We obtained 21-fs, 14-mJ pulses with an extraction efficiency of 32% in the final amplifier. The dispersion through the system was successfully compensated for by insertion of a pair of prisms in a regenerative amplifier. The pulses were characterized by frequency-resolved optical gating and were in good agreement with dispersion calculated by three-dimensional ray tracing.

157-nm coherent light source as an inspection tool for F 2 laser lithography

We have developed a 157-nm coherent light source by two-photon resonant four-wave mixing in Xe, with two tunable single-mode 1-kHz Ti:sapphire laser systems at 768 and 681 nm. This light source has been developed to determine the instrumental function of a vacuum ultraviolet spectrometer and to evaluate optical designs for ultra-line-narrowed F(2) laser lithography. The spectral linewidth of the source was less than 0.008 pm (FWHM), with an average power of 0.6 mW.

Differential responses of normal human coronary artery endothelial cells against multiple cytokines comparatively assessed by gene expression profiles

Endothelial cells play an important role in terms of biological functions by responding to a variety of stimuli in the blood. However, little is known about the molecular mechanism involved in rendering the variety in the cellular response. To investigate the variety of the cellular responses against exogenous stimuli at the gene expression level, we attempted to describe the cellular responses with comprehensive gene expression profiles, dissect them into multiple response patterns, and characterize the response patterns according to the information accumulated so far on the genes included in the patterns. We comparatively analyzed in parallel the gene expression profiles obtained with DNA microarrays from normal human coronary artery endothelial cells (HCAECs) stimulated with multiple cytokines, interleukin‐1β, tumor necrosis factor‐α, interferon‐β, interferon‐γ, and oncostatin M, which are profoundly involved in various functional responses of endothelial cells. These analyses revealed that the cellular responses of HCAECs against these cytokines included at least 15 response patterns specific to a single cytokine or common to multiple cytokines. Moreover, we statistically extracted genes contained within the individual response patterns and characterized the response patterns with the genes referring to the previously accumulated findings including the biological process defined by the Gene Ontology Consortium (GO). Out of the 15 response patterns in which at least one gene was successfully extracted through the statistical approach, 11 response patterns were differentially characterized by representing the number of genes contained in individual criteria of the biological process in the GO only. The approach to dissect cellular responses into response patterns and to characterize the pattern at the gene expression level may contribute to the gaining of insight for untangling the diversity of cellular functions.

Generation of milliwatt narrow-bandwidth vacuum ultraviolet radiation by an all-solid-state tunable high-average-power laser system

Generation of milliwatt narrow-bandwidth vacuum ultraviolet radiation by two-photon resonant four-wave mixing in Xe at 153 nm is demonstrated. The output of extreme ultraviolet radiation was at the microwatt level at 85 nm. For this demonstration, we developed an all-solid-state tunable 5-kHz Ti:sapphire laser system that produces 0.6-ns 0.7-GHz-bandwidth pulses at an average power of 32 W at the fundamental, 12 W at the second harmonic, and 6.3 W at the third harmonic.

Molecular cloning and characterization of a gene expressed in mouse developing tongue, mDscr5 gene, a homolog of human DSCR5 (Down syndrome Critical Region gene 5)

Abstract. For understanding the pathogenesis of Down syndrome (DS), it is important to identify and characterize the genes on Chromosome (Chr) 21, especially those in the Down syndrome critical region (DSCR) on Chr 21q22.2. Recently we have determined 33.5 Mb (more than 99%) of DNA sequence of Chr 21 and, from these sequence data, we identified a novel gene, DSCR5 (transcript = 0.8 kb), from DSCR by combination of computational gene prediction and cDNA screening. For functional analysis of DSCR5, we identified a mouse homolog of the DSCR5 cDNA, and termed it mDscr5 (transcript length = 0.8 kb). The gene was mapped to mouse Chr 16 C3-C4, the syntenic region of human Chr 21, and encodes an amino acid of 132 residues with 90% identity to DSCR5. In situ hybridization showed that mDscr5 is predominantly expressed in the developing tongue. To our best knowledge, no other gene in DSCR is reported to be expressed in tongue, so that DSCR5 may be the first candidate to elucidate the pathophysiology of tongue malformation observed in DS.

Second harmonic generation from KBBF crystal in the deep ultraviolet

By using a KBe/sub 2/BO/sub 3/F/sub 2/ (KBBF) crystal with a size of 10 /spl times/ 10 /spl times/ 1.2 mm/sup 3/ and a special prism coupling technique (PCT), the fourth-harmonic generation of Ti:sapphire laser systems from 200 nm to 179.4-nm has been achieved for the first time. Moreover, by a Ti:sapphire laser with a 50-fs pulse duration and a 1-kHz repetition rate. A conversion efficiency as high as 13% from 400 nm to 200 nm has also been obtained without any surface loss correction.

Molecular Cloning and Phylogenetic Analysis of Canine β-Casein

A canine β-casein cDNA was isolated from mammary tissue by polymerase chain reaction (PCR) using degenerate primers. It encodes 250 amino acids protein containing the conserved sequence motif of β-casein. It showed the highest homology with snow-leopard (Uncia uncia) (55–62 % identity). It also showed 44–5396 identity with human, 3342 % identity with mouse, 29–37 % identity with rat, 43–53 % identity with rabbit, 4148 % identity with pig, 44–51 % identity with cattle and 44–50 % identity with sheep. A 1.2-kb mRNA was detected in mammary tissue by Northern blot analysis. Phylogenetic analysis revealed that canine β-casein formed a branch with lesser panda and snow leopard, which were grouped into carnivore.

Construction of a novel cell-based assay for the evaluation of anti-EGFR drug efficacy against EGFR mutation

Epidermal growth factor receptor (EGFR) overexpression and EGFR-mediated signaling pathway dysregulation have been observed in tumors from patients with various cancers, especially non-small cell lung cancer. Thus, several anti-EGFR drugs have been developed for cancer therapy. For patients with known EGFR activating mutations (EGFR exon 19 in-frame deletions and exon 21 L858R substitution), treatment with an EGFR tyrosine kinase inhibitor (EGFR TKI; gefitinib, erlotinib or afatinib) represents standard first-line therapy. However, the clinical efficacy of these TKIs is ultimately limited by the development of acquired drug resistance such as by mutation of the gatekeeper T790 residue (T790M). To overcome this acquired drug resistance and develop novel anti-EGFR drugs, a cell-based assay system for EGFR TKI resistance mutant-selective inhibitors is required. We constructed a novel cell-based assay for the evaluation of EGFR TKI efficacy against EGFR mutation. To this end, we established non-tumorigenic immortalized breast epithelial cells that proliferate dependent on EGF (MCF 10A cells), which stably overexpress mutant EGFR. We found that the cells expressing EGFR containing the T790M mutation showed higher resistance against gefitinib, erlotinib and afatinib compared with cells expressing wild-type EGFR. In contrast, L858R mutant-expressing cells exhibited higher TKI sensitivity. The effect of T790M-selective inhibitors (osimertinib and rociletinib) on T790M mutant-expressing cells was significantly higher than gefitinib and erlotinib. Finally, when compared with commercially available isogenic MCF 10A cell lines carrying introduced mutations in EGFR, our EGFR mutant-overexpressing cells exhibited obviously higher responsiveness to EGFR TKIs depending on the underlying mutations because of the higher levels of EGFR phosphorylation in the EGFR mutant-overexpressing cells than in the isogenic cell lines. In conclusion, we successfully developed a novel cell-based assay for evaluating the efficacy of anti-EGFR drugs against EGFR mutation.

Generation of vacuum-ultraviolet light by an optical contact, prism-coupled KBBF crystal

We generated the second harmonic of a frequency-tripled Nd:YVO/sub 4/ laser with an quasi-CW, 2.5-mW output and also the shortest-wavelength second harmonic at 172.5 nm and sum-frequency mixing at 163.3 nm by an optical contact, prism-coupled KBe/sub 2/BO/sub 3/F/sub 2/ (KBBF) crystal.