Retention of the developmental pluripotency in medaka embryonic stem cells after gene transfer and long-term drug selection for gene targeting in fish

Embryonic stem (ES) cells provide a unique tool for introducing random or targeted genetic alterations, because it is possible that the desired, but extremely rare recombinant genotypes can be screened by drug selection. ES cell-mediated transgenesis has so far been limited to the mouse. In the fish medaka (Oryzias latipes) several ES cell lines have been made available. Here we report the optimized conditions for gene transfer and drug selection in the medaka ES cell line MES1 as a prelude for gene targeting in fish. MES1 cells gave rise to a moderate to high transfection efficiency by the calcium phosphate co-precipitation (5%), commercial reagents Fugene (11%), GeneJuice (21%) and electroporation (>30%). Transient gene transfer and CAT reporter assay revealed that several enhancers/promoters and their combinations including CMV, RSV and ST (the SV40 virus early gene enhancer linked to the thymidine kinase promoter) were suitable regulatory sequences to drive transgene expression in the MES1 cells. We show that neo, hyg or pac conferred resistance to G418, hygromycin or puromycin for positive selection, while the HSV-tk generated sensitivity to ganciclovir for negative selection. The positive-negative selection procedure that is widely used for gene targeting in mouse ES cells was found to be effective also in MES1 cells. Importantly, we demonstrate that MES1 cells after gene transfer and long-term drug selection retained the developmental pluripotency, as they were able to undergo induced differentiation in vitro and to contribute to various tissues and organs during chimeric embryogenesis.

Growth performance of juvenile white sturgeon as affected by feeding regimen

Juvenile (mean +/- SE, 8.6 +/- 0.1 g) white sturgeon Acipenser transmontanus were fed for 8 weeks under one of six feeding regimens: continuously 24 h/d (C24); continuously 12.8 h/d during the day (C12/D), continuously 12.8 h/d at night (C12/N), 6 meals/d (M6), 4 meals/d (M4), and 2 meals/d (M2). Specific growth rate, feed efficiency, and body lipid content were significantly (P < 0.05) affected by the feeding regimen. These variables were highest in the C24 group and lowest in the M2 group; fish in the M6 group showed the second best performance. Specific growth rate and feed efficiency in terms of wet weight in the M6 groups were not significantly different from those in the C24 groups, but specific growth rate in terms of energy and energy retention efficiency were significantly lower. Feeding regimen had no effect on condition factor, hepatosomatic index, coefficient of variation in final body weight, and protein and ash contents. There was no significant difference in these indexes between 12.8-h/d continuous feeding by day or by night. It was concluded that continuous feeding for 24 h/d was the optimum feeding regimen for juvenile white sturgeon.

Fabrication of a Low-Loss SSC Using High-Dose Electron Beam Lithography Exposure With Negative PMMA Resist

A silicon-on-insulator optical fiber-to-waveguide spot-size converter (SSC) using Poly-MethylMethAcrylate (PMMA) is presented for integrated optical circuits. Unlike the conventional use of PMMA as a positive resist, it has been successfully used as a negative resist with high-dose electron exposure for the fabrication of ultrafine silicon wire waveguides. Additionally, this process is able to reduce the side-wall roughness, and substantially depresses the unwanted propagation loss. Exploiting this technology, the authors demonstrated that the SSC can improve coupling efficiency by as much as over 2.5 dB per coupling facet, compared with that of SSC fabricated with PMMA as a positive resist with the same dimension.

Effects of indium ion implantation on the total dose hardness of fully depleted SOI NMOSFET

In this work, we investigate the effects of the indium ion implantation towards the back-channel interface on the total dose hardness of the n-channel SOI MOSFET. The results show that the indium implant has slight impact on the normal threshold voltage while preserving low leakage current after irradiation. The advantage is attributed to the narrow as-implanted and postanneal profile of the indium implantation. Two-dimensional simulations have been used to understand the physical mechanisms of the effects.

The impact of implantation dose on the characteristics of diluted-magnetic nonpolar GaN:Cu films

Diluted-magnetic nonpolar GaN:Cu films have been fabricated by implanting Cu ions into p-type nonpolar a-plane (1120) GaN films with a subsequent thermal annealing process. The impact of the implantation dose on the structural. morphological and magnetic characteristics of the samples have been investigated by means of high-resolution X-ray diffraction (HRXRD). atomic force microscopy (AFM), and superconducting quantum interference device (SQUID). The XRD and AFM analyses show that the structural and morphological characteristics of samples deteriorated with the increase of implantation dose. According to the SQUID analysis. obvious room-temperature ferromagnetic properties of samples were detected. Moreover, the saturation magnetization per Cu atom decreased as the implantation dose increased. (C) 2009 Elsevier B.V. All rights reserved.

Silicon-on-insulating multi-layers for total-dose irradiation hardness

Silicon-on-insulating multi-layer (SOIM) materials were fabricated by co-implantation of oxygen and nitrogen ions with different energies and doses. The multilayer microstructure was investigated by cross-sectional transmission electron microscopy. P-channel metal-oxide-semiconductor (PMOS) transistors and metal-semiconductor-insulator-semiconductor (MSIS) capacitors were produced by these materials. After the irradiated total dose reaches 3 x 10(5) rad (Si), the threshold voltage of the SOIM-based PMOS transistor only shifts 0.07 V, while thin silicon-on-insulating buried-oxide SIMOX-based PMOS transistors have a shift of 1.2V, where SIMOX represents the separated by implanted oxygen. The difference of capacitance of the SOIM-based MSIS capacitors before and after irradiation is less than that of the thin-box SIMOX-based MSIS capacitor. The results suggest that the SOIM materials have a more remarkable irradiation tolerance of total dose effect, compared to the thin-buried-oxide SIMOX materials.

Formation of total-dose-radiation hardened materials by sequential oxygen and nitrogen implantation and multi-step annealing

Separation by implantation of oxygen and nitrogen (SIMON) silicon-on-insulator (SOI) materials were fabricated by sequential oxygen and nitrogen implantation with annealing after each implantation. Analyses of SIMS, XTEM and HRTEM were performed. The results show that superior buried insulating multi-layers were well formed and the possible mechanism is discussed. The remarkable total-dose irradiation tolerance of SIMON materials was confirmed by few shifts of drain leakage current-gate source voltage (I-V) curves of PMOS transistors fabricated on SIMON materials before and after irradiation.

Fabrication of metallic air bridges using multiple-dose electron beam lithography

The techniques of fabricating metallic air bridges using different resists in a one-step electron beam lithography are presented. The exposure process employed a single-layer polymethyl methacrylate (PMMA) or photoresists with either different doses in the span and feet areas or with varying acceleration voltage of the electron beam. The process using photoresists with different doses has produced air bridges more stable than what the PMMA method using various acceleration voltages would achieve. Using this method, air bridges up to 12 mu m long have been fabricated. The length and height of these metallic air bridges vary with the photoresist thickness. (c) 2006 American Institute of Physics.

Dose rate dependence of electrical characteristics of lead zirconate titanate capacitors

The influence of gamma-radiation dose rate on the electrical properties of lead zirconate titanate capacitors was investigated. More severe degradations in dielectric constant, coercive field, remanent polarization and capacitance-voltage (C-V) curves occurred with increasing radiation dose at lower dose rates. The electrical properties exhibited distinct radiation dose rate dependence and the worst-case degradation occurred at the lowest dose rate. The radiation-induced degradation of parameters such as the coercive field drift and distortion of the C-V curve can be recovered partly through post-irradiation annealing.

Aggregation and precipitation in high dose as ion implanted Ge0.5Si0.5 alloy

Thermally stimulated redistribution and precipitation of excess arsenic in Ge0.5Si0.5 alloy has been studied by X-ray photoelectron spectroscopy (XPS), cross sectional transmission electron microscopy (XTEM) and X-ray energy disperse spectrometry (EDS). Samples were prepared by the implantation of 6 X 10(6) As+ cm(-2) and 100 keV with subsequent thermal processing at 800 degrees C and 1000 degrees C for 1 h. The XPS depth profiles from the implanted samples before and after the thermal annealing indicate that there is marked redistribution of the elements in heavily arsenic-implanted Ge0.5Si0.5 alloys during the annealing, including: (1) diffusion of As from the implanted region to the surface; (2) aggregation of Ge in the vicinity of the surface. A high density of precipitates was observed near the surface which were by XTEM and EDS identified as an arsenide. It is suggested that most of the implanted As in Ge0.5Si0.5 alloy exists in the form of GeAs.

Total Dose Radiation-Hard 0. SOI CMOS Transistors and ASIC

This paper presents the total dose radiation performance of 0. S^m SOI CMOS devices fabricated with full dose SIMOX technology. The radiation performance is characterized by threshold voltage shifts and leakage currents of transistors and standby currents of ASIC as functions of the total dose up to 500krad(Si) .The experimental results show that the worst case threshold voltage shifts of front channels are less than 320mV for pMOS transistors under off-gate radiation bias at lMrad(Si) and less than 120mV for nMOS transistors under on-gate radiation bias. No significant radiation-induced leakage current is observed in transistors to lMrad(Si). The standby currents of ASIC are less than the specification of 5μA over the total dose range of 500krad(Si).