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Production of EGFP and dsRed Expressing Transgenic Mice
Hideaki Yamashiro,Stefan Moisyadi 한국동물생명공학회(구 한국동물번식학회) 2011 발생공학 국제심포지엄 및 학술대회 Vol.2011 No.1
<Objective> Injection of a linear transgene into male pronucleus has been widely used to produce Transgenic (Tg) mice. This approach however is inefficient and results in concatemerised transgene insertion and associated reduced protein expression from such insertions. The objective of this study is to develop active transgenesis method by using a piggyBac transposase plasmid DNA, and generate double transgene harboring transgenic mice. <Method> We examined the piggyBac transposase plasmid (pm- GENIE‐3) on its ability to produce transgenic animals with NaOH, HCl and FuGENE6 treated sperm followed by ICSI‐Tr, for its effectiveness in creating EGFP Tg mice, as judged by offspring epifluorescence. After these steps, we explored if the embryo development affects ICSI‐Tr efficiency by using substrate‐free media or aphidicolin. Moreover, we tested to determine if transgenesis is possible by directly injecting the DNA into the cytoplasm or into pronuclei. Finally, we attempted the introduction of two transgenes, such as EGFP and dsRED simultaneously in one transposon and the ability to generate double Tg mice by using NaOH treated sperm during ICSI‐Tr. <Results> The best results were obtained when sperm were treated with NaOH and co‐incubated with circular plasmid DNA of pmGENIE‐3. This resulted in Tg pups that could successfully express EGFP, with efficiencies of 37.9% of born animals being transgenic. Furthermore, the effectiveness of this method was proved by the production of Tg offspring from inbred strains of mice, such as C57BL/6, Balb/c and CD‐1 nude. While injection of DNA into the pronucleus or cytoplasm of one cell embryos, and delayed embryo development‐method were not as effective as ICSI‐Tr in producing Tg mice, they nevertheless proved successful. Finally, NaOH‐ICSI‐Tr successfully obtained Tg mice expressing both the EGFP and dsRED transgene. In conclusion, the current study developed an active form of NaOH‐ICSI‐Tr mediated transgenesis utilizing the piggyBac transposition machinery, and was successful in obtaining Tg mice which expressed simultaneously not only EGFP but also the dsRED transgene stably inserted in these animals.
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Miyayama Shiro,Yamashiro Masashi,Ikeda Rie,Yokka Akira,Komiya Hideaki,Sakuragawa Naoko 소화기인터벤션의학회 2023 International journal of gastrointestinal interven Vol.12 No.2
Background: To evaluate the outcomes of conventional transarterial chemoembolization (TACE) using guidance software for hepatocellular carci-noma (HCC) in the caudate lobe.Methods: Seventy-two patients with 79 treatment-naïve caudate lobe HCCs were eligible. TACE was performed through feeders not only identified by automated tumor-feeder detection (AFD) functionality but also detected manually. Technical success of TACE was classified into three grades accord-ing to 1-week computed tomography findings: entire tumor embolized with a safety margin (5 mm for tumors < 25 mm and 10 mm for tumors ≥ 25 mm) (grade A); tumor embolized without a margin in parts (grade B); and entire tumor not embolized (grade C). Tumor response was evaluated using the modified Response Evaluation Criteria in Solid Tumors. Tumor-feeder detectability by AFD, technical success of TACE, complete response (CR) at 2–4 months, durable CR, and local tumor progression (LTP) rates calculated by the Kaplan-Meier method were compared in each tumor among three subsegments: the Spiegel lobe (SP); paracaval portion (PC); and caudate process (CP). LTP rates between the grade A and B tumors were also com-pared.Results: The mean tumor diameter was 18.6 ± 9.9 mm (range, 6–53 mm), and 111 of 129 (86.0%) tumor feeders were detected by AFD. The rates of feeder detectability by AFD, grade A technical success of TACE, CR at 2–4 months, and durable CR in 30 SP, 38 PC, and 11 CP tumors were 71.4%, 93.3%, 93.1%, and 79.3%; 94.8%, 65.8%, 59.4%, and 34.4%; and 76.5%, 63.6%, 80.0%, and 30.0%, respectively. LTP rates of SP tumors were sig-nificantly lower than those of PC tumors (P = 0.0044), and the grade B tumors progressed more frequently (P = 0.0012).Conclusion: AFD could detect 86.0% of tumor feeders in the caudate lobe; however, the feeder detectability, technical success of TACE, and outcomes differed among the three subsegments.
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Magnetic and charge orders in zigzag nanographene ribbons
Kikuo Harigaya,Atsushi Yamashiro,Yukihiro Shimoi,Katsunori Wakabayashi 한국물리학회 2004 Current Applied Physics Vol.4 No.6
We theoretically study the electronic states in graphene ribbons which are strongly aected by the edge states, the peculiar non-bonding molecular orbitals localized along the zigzag edges of the ribbons. New kinds of edge localized electronic states with spinand charge polarizations are found in the mean eld solutions of the extended Hubbard model with onsite and nearest-neighborCoulomb repulsions. These novel states appear due to the interplay between the edge states and the Fermi instabilities. We alsoexamine the competition between the charge polarized state and the spin polarized state to draw a phase diagram depending onCoulomb parameters. The results obtained by the mean eld calculations with the extended Hubbard model modied to include Coulomb integrals provide useful insights to understand and functionalize the nanoscale materials.
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Shiro Miyayama,Masashi Yamashiro,Rie Ikeda,Junichi Matsumoto,Nobuhiko Ogawa,Kazuo Notsumata 소화기인터벤션의학회 2022 Gastrointestinal Intervention Vol.11 No.1
Background: Bleeding from duodenal varices is a rare but life-threatening complication of portal hypertension. The treatment of duodenal varices remains difficult and a definitive treatment strategy has not been established. The aim of this study was to report the technical aspects and outcomes of balloon-occluded retrograde transvenous obliteration (BRTO) using 5% ethanolamine oleate with iopamidol (EOI) for duodenal varices. Methods: Six consecutive patients with duodenal varices treated using BRTO were eligible. Endoscopic treatment was performed first in three patients with active bleeding. After coil embolization of collateral veins, stepwise EOI infusion was performed at intervals of 10–30 minutes under balloon occlusion until the main efferent vein, varices, and the main afferent vein were filled with EOI and clots. The techniques and outcomes of BRTO were retrospectively evaluated. Results: The main efferent vein of duodenal varices was the right (n = 4) or left (n = 2) gonadal vein. In three patients with ruptured varices, BRTO was performed after achieving hemostasis by endoscopic treatment. In five patients, 1–4 (mean, 2.4 ± 1.1) collateral veins were embolized with coils before EOI infusion. Furthermore, 11–21 mL (mean, 15.3 ± 4.2 mL) of EOI was infused by 3–5 (mean, 3.5 ± 1.0) stepwise infusions via the efferent vein under balloon occlusion. The duration of EOI infusion under balloon occlusion ranged from 82 to 118 minutes (mean, 87.8 ± 13.6 minutes). The varices were thrombosed in all but one patient. In the remaining patient, the varices were thrombosed by additional BRTO under overnight balloon occlusion performed 19 days later. The only complications were a transient fever and hematuria. All duodenal varices disappeared during a followup of 4–32 months (mean, 16.2 ± 11.1 months) after BRTO. Conclusion: BRTO using EOI is an effective treatment for duodenal varices.
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Examining the Physical Properties and Implications of Tuffisite Presence in Singapore
( Keisuke Tada ),( Yoshiro Yamashiro ),( Sing Lih Chiam ),( Yong Siang Lim ) 대한지질공학회 2019 대한지질공학회 학술발표회논문집 Vol.2019 No.2
It has been recently discovered that tuffisites distribute in the Bukit Timah Granite and Gombak Norite in central Singapore. Tuffisites are intrusive fragmental igneous rocks found in intrusive fractures formed by the explosive release of gas, highly pressurised hydrothermal fluid or magma. As tuffisites consist of brecciated and lithified fine to coarse grained rock fragments, which present different physical properties from the country rock, the distribution and physical properties of tuffisites can possess a significant impact on engineering properties of the rock mass in the plutons. The study summarises the distribution, condition, results of petrographic analysis, laboratory test and in-situ test of tuffisite observed at the Bukit Timah Granite and Gombak Norite in Singapore. Tuffisites are identified by petrographic analysis at four boreholes in depths from 37.0 to 172.0 metres and observed based on visual inspection at other four boreholes in depths from 15.7 to 150.0 metres. The distribution of tuffisites widely spreads across the plutons, and their thickness ranges from 0.1 to 134.3 metres. Unconfined compressive strength test and point load test results are summarised and assessed in terms of lithified condition of tuffisites. Tuffisites present relatively low strength compare to the country rock although the strengths vary with level of lithification, which implies that sudden strength change may be encountered in deep excavation at the plutons. Results of Lugeon test, porosity test, slake durability test and jointing degree are summarised and discussed to understand more about the mechanical properties of tuffisites.
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Shiro Miyayama,Masashi Yamashiro,Rie Ikeda,Junichi Matsumoto,Nobuhiko Ogawa,Kazuo Notsumata 소화기인터벤션의학회 2022 International journal of gastrointestinal interven Vol.11 No.1
Background: Bleeding from duodenal varices is a rare but life-threatening complication of portal hypertension. The treatment of duodenal varices remains difficult and a definitive treatment strategy has not been established. The aim of this study was to report the technical aspects and outcomes of balloon-occluded retrograde transvenous obliteration (BRTO) using 5% ethanolamine oleate with iopamidol (EOI) for duodenal varices. Methods: Six consecutive patients with duodenal varices treated using BRTO were eligible. Endoscopic treatment was performed first in three patients with active bleeding. After coil embolization of collateral veins, stepwise EOI infusion was performed at intervals of 10–30 minutes under balloon occlusion until the main efferent vein, varices, and the main afferent vein were filled with EOI and clots. The techniques and outcomes of BRTO were retrospectively evaluated. Results: The main efferent vein of duodenal varices was the right (n = 4) or left (n = 2) gonadal vein. In three patients with ruptured varices, BRTO was performed after achieving hemostasis by endoscopic treatment. In five patients, 1–4 (mean, 2.4 ± 1.1) collateral veins were embolized with coils before EOI infusion. Furthermore, 11–21 mL (mean, 15.3 ± 4.2 mL) of EOI was infused by 3–5 (mean, 3.5 ± 1.0) stepwise infusions via the efferent vein under balloon occlusion. The duration of EOI infusion under balloon occlusion ranged from 82 to 118 minutes (mean, 87.8 ± 13.6 minutes). The varices were thrombosed in all but one patient. In the remaining patient, the varices were thrombosed by additional BRTO under overnight balloon occlusion performed 19 days later. The only complications were a transient fever and hematuria. All duodenal varices disappeared during a followup of 4–32 months (mean, 16.2 ± 11.1 months) after BRTO. Conclusion: BRTO using EOI is an effective treatment for duodenal varices.
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