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Now showing items 1 - 6 of 6

  • Surge Analysis on a Long Underground Cable System

    Kawamura, Kazuki   Ametani, Akihiro   Gudmundsdottir, Unnur Stella  

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  • Formation of highly charged nanodroplets by condensation-electrospray device

    Seto, Takafumi   Maekawa, Tetsuya   Osone, Saho   Kawamura, Kazuki   Yamauchi, Toshiyuki   Otani, Yoshio  

    A novel electrospray device is developed to generate highly charged nanodroplets from dew-condensed water. The device is composed of a discharge electrode with a spherical tip, ground electrode, Peltier cooling device, and high voltage power supply. When the surface temperature of the discharge electrode reaches the dew point of surrounding room air, a liquid water film is formed on the discharge electrode. Pulsating cone-jet mode electrospray is generated by applying negative 5 kV high voltage to the discharge electrode. Highly charged macrodroplets are ejected from the tip of water cone and they are externally charged by the collision of unipolar negative ions. As the droplets shrink by the evaporation while keeping the number of charges, the droplets undergo Coulombic fission and/or electron emission when the number of charges exceeds the charge retention limits. The resulting change in the electrical mobility distribution and the number of charge are measured by the aerosol measurement techniques. It is found that droplets carry 3 to 23 elementary charges and the charge distribution has a peak at n=8. (C) 2012 Elsevier B. V. All rights reserved.
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  • Inactivation of Sonic Hedgehog Signaling and Polydactyly in Limbs of Hereditary Multiple Malformation,a Novel Type of Talpid Mutant

    Matsubara, Yoshiyuki   Nakano, Mikiharu   Kawamura, Kazuki   Tsudzuki, Masaoki   Funahashi, Jun-Ichi   Agata, Kiyokazu   Matsuda, Yoichi   Kuroiwa, Atsushi  

    Hereditary Multiple Malformation (HMM) is a naturally occurring, autosomal recessive, homozygous lethal mutation found in Japanese quail. Homozygote embryos (hmm(-/-)) show polydactyly similar to talpid(2) and talpid(3) mutants. Here we characterize the molecular profile of the hmm(-/-) limb bud and identify the cellular mechanisms that cause its polydactyly. The hmm(-/-) limb bud shows a severe lack of sonic hedgehog (SHH) signaling, and the autopod has 4 to 11 unidentifiable digits with syn-, poly-, and brachydactyly. The Zone of Polarizing Activity (ZPA) of the hmm(-/-) limb bud does not show polarizing activity regardless of the presence of SHH protein, indicating that either the secretion pathway of SHH is defective or the SHH protein is dysfunctional. Furthermore, mesenchymal cells in the hrnm(-/-) limb bud do not respond to ZPA transplanted from the normal limb bud, suggesting that signal transduction downstream of SHH is also defective. Since primary cilia are present in the hmm(-/-) limb bud, the causal gene must be different from talpid(2) and talpid(3). In the hmm(-/-) limb bud, a high amount of GLI3A protein is expressed and GLI3 protein is localized to the nucleus. Our results suggest that the regulatory mechanism of GLI3 is disorganized in the hmm(-/-) limb bud.
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  • Primary Realignment for Pelvic Fracture Urethral Injury Is Associated With Prolonged Time to Urethroplasty and Increased Stenosis Complexity

    Horiguchi, Akio   Shinchi, Masayuki   Masunaga, Ayako   Okubo, Kazuki   Kawamura, Kazuki   Ojima, Kenichiro   Ito, Keiichi   Asano, Tomohiko   Azuma, Ryuichi  

    OBJECTIVE To compare the clinical courses of patients with pelvic fracture urethral injury (PFUI) according to initial management strategy. METHODS We reviewed the clinical courses of 63 patients with PFUI who were initially treated elsewhere and underwent delayed anastomotic urethroplasty by a single surgeon between 2008 and 2015. Patients were grouped according to their initial treatment: by suprapubic tube placement alone (49 patients, SPT group) or primary realignment (14 patients, PR group). Time to urethroplasty was defined as the period between injury and delayed urethroplasty. Clinical data regarding the status of urethral stenosis, urethroplasty procedure, and treatment outcome were analyzed. RESULTS The mean time to urethroplasty in the PR group was about 3 times than that in the SPT group (133 months vs 47 months, P =3D .035). Fifty percent of the PR group (7 of 14) had a history of repeated urethrotomy or dilation before referral, a percentage significantly higher than that of the SPT group (20.4%, 10 of 49, P =3D .027). The percentage of patients having a false passage and iatrogenic scar was significantly higher in the PR group (42.9% vs 16.3%, P =3D .035), but there was no significant between-group difference in urethral stenosis length, operative time, operative blood loss, or the percentage of patients requiring inferior pubectomy or urethral rerouting. CONCLUSION PR does not facilitate delayed urethroplasty, and patients who undergo PR are at high risk of having a more complicated stenosis and longer time to urethroplasty, presumably because of repeated transurethral procedures. (C) 2017 Elsevier Inc.
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  • Identification of transgene integration site and anatomical properties of fluorescence intensity in a EGFP transgenic chicken line

    Tsujino, Kaori   Okuzaki, Yuya   Hibino, Nobuyuki   Kawamura, Kazuki   Saito, Seiji   Ozaki, Yumi   Ishishita, Satoshi   Kuroiwa, Atsushi   Iijima, Shinji   Matsuda, Yoichi   Nishijima, Kenichi   Suzuki, Takayuki  

    Transgenic birds are commonly used for time-lapse imaging and fate mapping studies in developmental biology. When researchers use transgenic birds expressing fluorescent protein, they need to understand the integration site of the transgene in the genome and the intensity of fluorescence in the tissues of interest. In this study, we determined the integration site of the transgene and fluorescence property of developing organs in our transgenic chicken line generated by lentivirus infection. The transgene was localized between exons 3 and 4 of MED27. Some homozygotes and heterozygotes appeared to be lethal at early embryonic stages. We performed histological analysis of EGFP expression in transgenic embryos at St. 14, 17, and 24 by immunohistochemistry with anti-GFP antibody on paraffin sections. Next, we cut cryosections and quantified direct EGFP intensity from the transgene in each tissue without performing immunohistochemistry. These results revealed that EGFP intensity in each tissue was unique in developing embryos and changed according to developmental stages. Finally, we demonstrated that EGFP-expressing cells in a micromass culture with co-culturing wild-type cells were clearly distinguishable via live cell imaging. These results provide essential information on the potential of our transgenic line and indicate that these transgenic chicken lines are useful for research associated with developmental biology.
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  • Brilliant Blue as an alternative dye to Fast Green for in ovo electroporation

    Saito, Seiji   Kawamura, Kazuki   Matsuda, Yoichi   Suzuki, Takayuki  

    Chick embryo electroporation is a powerful tool for the introduction of transgenes into tissues of interest for the study of developmental biology. This method often uses Fast Green to visualize the injected area by staining the solution containing DNA green. Here, we show that Fast Green fluoresces in a red color after electroporation, suggesting that researchers need to be cautious when detecting red fluorescence. Fast Green solution did not show any fluorescence before injection into chick embryos, but fluoresced red within 3 min post-injection into chick embryos. We identified Brilliant Blue as suitable alternative dye for use as an indicator of injection sites in ovo electroporation. We found that 0.2% of Brilliant Blue was sufficient to track the area of DNA injection. In addition, this chemical did not show red fluorescence after electroporation. Our findings demonstrate that Brilliant Blue can be used for detecting red fluorescent proteins introduced into chick embryos by electroporation. Our study also shows useful examples for the application of Brilliant Blue for the precise quantification of two fluorescence intensities after EGFP and mCherry co-electroporation.
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