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151.
Obayashi H Kawabe Y Makitsubo H Watanabe R Kameyama Y Huang S Takenouchi Y Ito A Kamihira M 《Journal of Bioscience and Bioengineering》2012,113(3):381-388
Site-specific gene recombination systems, such as Cre/loxP, have been used for genetic modification of cells and organisms in both basic and applied research. We previously developed an accumulative gene integration system (AGIS), in which target gene cassettes could be repeatedly integrated into a pre-determined site on a plasmid or cellular genome by recombinase-mediated cassette exchange (RMCE), using Cre and mutated loxPs. In the present study, we designed a simplified AGIS. For gene integration into a target site, the previous system used two loxP sites in the acceptor DNA, whereas the new system uses a single loxP site. The gene integration reactions were repeated four times in vitro using Cre protein and specific plasmids. The expected integration reactions mediated by Cre occurred at the loxP sites, resulting in integration of four target genes. The system was also used for genomic integration of reporter genes using Chinese hamster ovary (CHO) cells. The reporter genes were efficiently introduced into the CHO genome in a Cre-dependent manner, and transgene expression was detected after the integration reaction. The expression levels of the reporter genes were enhanced, corresponding to the increase of transgene copy number. Recombinase-mediated AGIS provides a useful tool for the modification of cellular genomes. 相似文献
152.
Double-walled carbon nanotubes (DWCNTs) with high graphitization have been synthesized by hydrogen arc discharge. The obtained
DWCNTs have a narrow distribution of diameters of both the inner and outer tubes, and more than half of the DWCNTs have inner
diameters in the range 0.6–1.0 nm. Field electron emission from a DWCNT cathode to an anode has been measured, and the emission
current density of DWCNTs reached 1 A/cm2 at an applied field of about 4.3 V/μm. After high-temperature treatment of DWCNTs, long linear carbon chains (C-chains) can
be grown inside the ultra-thin DWCNTs to form a novel C-chain@DWCNT nanostructure, showing that these ultra-thin DWCNTs are
an appropriate nanocontainer for preparing truly one-dimensional nanostructures with one-atom-diameter.
相似文献
153.
Yoshinori Okamoto Hideto Jinno Shinji Itoh Shinya Shibutani 《International journal of molecular sciences》2021,22(13)
Human estrogens prescribed for hormone replacement therapy (HRT) are known to be potent carcinogens. To find safer estrogens, several chlorinated estrogens were synthesized and their carcinogenic potential were determined. A pellet containing either 2-chloro-17β-estradiol (2-ClE2) or 4-chloro-17β-estradiol (4-ClE2) was implanted subcutaneously for 52 weeks into August Copenhagen Irish (ACI) rats, a preferred animal model for human breast cancer. 17β-Estradiol (E2) frequently induced mammary tumors while both 2-ClE2 and 4-ClE2 did not. Their 17α-ethinyl forms, thought to be orally active estrogens, were also synthesized. Neither 2-chloro-17α-ethinylestradiol (2-ClEE2) nor 4-chloro-17α-ethinylestradiol (4-ClEE2) induced tumors. The less carcinogenic effects were supported by histological examination of mammary glands of ACI rats treated with the chlorinated estrogens. A chlorine atom positioned at the 2- or 4-position of E2 may prevent the metabolic activation, resulting in reducing the carcinogenicity. 2-ClE2 and 4-ClE2 administered subcutaneously and 2-ClEE2 and 4-ClEE2 given orally to ovariectomized rats all showed uterotrophic potency, albeit slightly weaker than that of E2. Our results indicate that less carcinogenic chlorinated estrogens retaining estrogenic potential could be safer alternatives to the carcinogenic estrogens now in use for HRT. 相似文献
154.
Heterotypic 3-D coculture is essential to mimic tissues and organs, because cell-cell interaction between various types of cells is believed to be important for the activation of cellular functions. In this study, magnetic force was applied to construct a 3-D coculture system of HepG2 and NIH3T3 cells as a model of hepatocytes and mesenchymal cells. Magnetite cationic liposomes (MCLs) were used to label target cells. NIH3T3 cells labeled with MCLs were seeded onto ultralow-attachment plates, whose surface is composed of a covalently bound hydrogel layer that is hydrophilic and neutrally charged. When a magnet was placed under the plate, cells accumulated on the bottom of the well. After a 24-h incubation period, the cells formed a multilayered cell sheet, which contained the major mesenchymal extracellular matrix (ECM) components (fibronectin and type I collagen), suggesting that the use of stromal NIH3T3 cells gave sufficient strength to cell sheets. Both NIH3T3 and HepG2 cells were labeled with MCLs, and cocultured by two methods: NIH3T3 cell sheets were constructed and HepG2 cells were subsequently seeded onto NIH3T3 cell sheets, and then allowed to form layered cell sheets by applying magnetic force; or NIH3T3 and HepG2 cells were mixed and then allowed to form mixed cell sheets by applying magnetic force. These heterotypic multilayered cell sheets were successfully constructed and an enhanced albumin secretion by HepG2 cells was observed. These results suggest that the new tissue engineering technique using magnetite nanoparticles and magnetic force, to which we refer to as magnetic force-based tissue engineering (Mag-TE), is a promising approach to construct multilayered cell sheets consisting of heterotypic cocultured cells. 相似文献
155.
Shimizu T Iijima K Miyabayashi K Ogawa Y Miyazaki H Sasada H Sato E 《Reproduction (Cambridge, England)》2007,134(5):677-682
Vascular endothelial growth factor (VEGF) expression in granulosa cells is associated with the thecal vasculature growth during ovarian follicular development. We hypothesized that injection of VEGF gene fragments directly into the rat ovary would induce production of a large number of ovulatory follicles and that these follicles would ovulate. To test this hypothesis, we treated immature female rats with combinations of hormones and VEGF gene fragments. The animals were divided into two groups: one group received solution containing transfection reagents as a control (n = 5), while the other group received direct ovarian injection of VEGF gene fragments at 19 (n = 5), 21 (n = 5), 23 (n = 5), or 25 (n = 5) days after birth followed by i.p. administration of 20 IU equine chorionic gonadotropin (eCG) at the age of 26 days. Forty-eight hours after eCG injection, animals were given 20 IU human chorionic gonadotropin (hCG) i.p. and then the oocytes in both groups were counted. The maximum number of ovulated oocytes was obtained when the VEGF gene fragments were injected into the rat ovary at 21 days after birth. Histological examination revealed that the injection of VEGF gene fragments markedly increased the vascular density around the preovulatory follicles and also the number of these follicles. Our data provide the first reported evidence that most ovulatory follicles generated by injection of VEGF gene fragments are able to ovulate upon hCG treatment. These results demonstrate that injection of VEGF gene fragments directly into the ovary stimulates the development of antral follicles by inducing the formation of thecal vasculature in immature female rats. 相似文献
156.
157.
Under intense UV light irradiation with an Nd:YAG laser at 355 nm, a gaseous mixture of iron pentacarbonyl (Fe(CO)5) and trimethylsilyl azide (TMSAz) produced aerosol magnetic particles with diameters of 50–100 nm. Two photon absorption of TMSAz and Fe(CO)5 took place efficiently to produce trimethylsilyl nitrene and iron carbonyls (Fe(CO) n n = 1–4). In cases where a partial pressure of TMSAz was high, nucleation reactions of aerosol particles proceeded via chemical reactions between trimethylsilyl nitrene and two iron carbonyls, resulting in bridging between two iron carbonyls by silyl nitrene. In cases where a partial pressure of TMSAz was low, the nucleation reactions proceeded via chemical reactions between two iron carbonyls, resulting in the formation of di-iron carbonyls with bridging >C?O group. Both of these nucleation reactions took place concurrently during aerosol particle formation. From FT-IR spectral change of sedimentary particles produced under an external magnetic field of up to 5 T, it was found that application of an external magnetic field accelerated the latter chemical reaction to result in the appearance of bridging ν(>C?O) band in FT-IR spectrum of the magnetic particles. Copyright 2013 American Association for Aerosol Research 相似文献
158.
In this article we propose the adaptive learning algorithm of neural network with respect to a rapid temperature change of forecasted day. The proposed adaptive learning algorithm is used to shift the learning range of previous year of forecasted day. Therefore, the proposed neural network can be trained by using learning data, including the maximum temperature to be forecasted. The suitability of the proposed approach is illustrated through an application to actual load data of Okinawa Electric Power Company in Japan. 相似文献
159.
Akira Taniguchi Tomoki Akita Kazuaki Yasuda Yoshinori Miyazaki 《International Journal of Hydrogen Energy》2008,33(9):2323-2329
The damage caused by cell reversal during proton exchange membrane fuel cells (PEMFCs) operation with air starvation was investigated by a single-cell experiment. Samples from degraded membrane–electrode assemblies (MEAs) were characterized. The loss of electrochemical surface area of the cathode platinum was detected by in situ cyclic voltammetry, and platinum sintering was detected by transmission electron microscopy (TEM) analysis. Degradation at the anode was not detected in the chemical analysis of the anode catalyst layer of MEA samples by energy dispersive X-ray analysis (EDX) and TEM. An obvious decrease in the performance of PEMFC was observed in a sample degraded by cell reversal for 120 min. 相似文献
160.
Hirotaka Tsuda Masahito Mori Yoshinori Takao Koji Eriguchi Kouichi Ono 《Thin solid films》2010,518(13):3475-3480
Formation mechanisms for profile anomalies such as surface roughness and residue have been investigated numerically and experimentally for Si etching in Cl2/O2 plasmas. The numerical simulation employed an atomic-scale cellular model (ASCeM) based on the Monte Carlo algorithm, which reproduced the feature profile evolution experimentally observed during etching at increased O2 concentrations. A comparison between simulation and experiment indicated that the local surface oxidation induces surface roughness at the bottom of the feature during etching; then, synergistic effects between surface oxidation and ion scattering in microstructural features on roughened surfaces increase the surface roughness, which in turn causes a number of significant residues or micropillars on bottom surfaces of the feature. In practice, in roughened microstructural features, geometrical shadowing effects for neutral oxygen suppress the surface oxidation at the local feature bottom, where the energetic ion incidence is increased owing to ion scattering on sidewalls of the local feature. 相似文献