Identification and characterization of a novel gene differentially expressed in zebrafish cross-subfamily cloned embryos

m Background: Cross-species nuclear transfer has been shown to be a potent approach to retain the genetic viability of a certain species near extinction. However, most embryos produced by cross-species nuclear transfer were compromised because that they were unable to develop to later stages. Gene expression analysis of cross-species cloned embryos will yield new insights into the regulatory mechanisms involved in cross-species nuclear transfer and embryonic development. Results: A novel gene, K31, was identified as an up-regulated gene in fish cross-subfamily cloned embryos using SSH approach and RACE method. K31 complete cDNA sequence is 1106 base pairs (bp) in length, with a 342 bp open reading frame (ORF) encoding a putative protein of 113 amino acids (aa). Comparative analysis revealed no homologous known gene in zebrafish and other species database. K31 protein contains a putative transmembrane helix and five putative phosphorylation sites but without a signal peptide. Expression pattern analysis by real time RT-PCR and whole-mount in situ hybridization (WISH) shows that it has the characteristics of constitutively expressed gene. Sub-cellular localization assay shows that K31 protein can not penetrate the nuclei. Interestingly, over-expression of K31 gene can cause lethality in the epithelioma papulosum cyprinid (EPC) cells in cell culture, which gave hint to the inefficient reprogramming events occurred in cloned embryos. Conclusion: Taken together, our findings indicated that K31 gene is a novel gene differentially expressed in fish cross-subfamily cloned embryos and over-expression of K31 gene can cause lethality of cultured fish cells. To our knowledge, this is the first report on the determination of novel genes involved in nucleo-cytoplasmic interaction of fish cross-subfamily cloned embryos.

Simultaneous determination of chromium(VI) and chromium(III) of bioavailable fraction in bottom mud by flow injection-atomic absorption spectroscopy

A rapid and sensitive method for separation and determination of Cr(VI) and Cr(III) in bottom mud of lake by flow injection on-line preconcentrtion system and GFAAS was developed. The available Cr(VI) and Cr(III) were extracted by HOAc or EDTA + NH4 NO3 and adsorbed simultaneously by an anion and a cation resin microclummn and then eluted simultaneously by 2 mol/L NH4 NO3 + 0.05 mol/L ascorbate and 2 mol/L H2SO4, respectively. The elution was performed for 50 s after adsorption for 2 min, and the efficiencies of elution were 85.4% - 94.8% and 96.7% - 106% for Cr(VI) and Cr(M) respectively. The detection limits of the method were 0.9 mu g/L and 2.7 mu g/L with relative standard deviations of 3.5% and 6.4% for the determination of Cr(VI) and Cr(III) in sample, respectively.

Excision of Sleeping Beauty transposons: parameters and applications to gene therapy

A major problem in gene therapy is the determination of the rates at which gene transfer has occurred. Our work has focused on applications of the Sleeping Beauty (SB) transposon system as a non-viral vector for gene therapy. Excision of a transposon from a donor molecule and its integration into a cellular chromosome are catalyzed by SB transposase. In this study, we used a plasmid-based excision assay to study the excision step of transposition. We used the excision assay to evaluate the importance of various sequences that border the sites of excision inside and outside the transposon in order to determine the most active sequences for transposition from a donor plasmid. These findings together with our previous results in transposase binding to the terminal repeats suggest that the sequences in the transposon-junction of SB are involved in steps subsequent to DNA binding but before excision, and that they may have a role in transposase-transposon interaction. We found that SB transposons leave characteristically different footprints at excision sites in different cell types, suggesting that alternative repair machineries operate in concert with transposition. Most importantly, we found that the rates of excision correlate with the rates of transposition. We used this finding to assess transposition in livers of mice that were injected with the SB transposon and transposase. The excision assay appears to be a relatively quick and easy method to optimize protocols for delivery of genes in SB transposons to mammalian chromosomes in living animals. Copyright (C) 2004 John Wiley Sons, Ltd.

Analysis of organchlorine compounds in water by solid phase microextraction and gas chromatography

In this work, both,solid phase microextraction (SPME) and solid phase extraction(SPE) were used to enrich organochlorine compounds in water samples and analyzed by gas chromatography with electron capture detector. The operating conditions of SPME have been studied and different kinds of solid phase were compared. Linear alkybenzene sulfonate(LAS) was added to the samples to investigate its effect on the analysis. The results indicated that polyacrylate was better than other commercial solid phases in extraction of moderated polar organic compounds and the sensitivity of SPME was higher than SPE. LAS affect much in liquid-liquid extraction and headspace SPME; but it has little effect on SPE and direct-SPME method. The applications showed that SPME was a fast and effective method in sample preparation.

Cleanup and quantification of polychlorinated dibenzo-p-dioxins/furans and polychlorinated biphenyls

By using modern techniques of isotope dilution, high-resolution gas chromatography/high-resolution mass spectrometry and multiple ions detection, an effective cleanup, qualitative and quantitative method was developed for polychlorinated dibenzo-p-dioxins/furans (PCDD/F) and polychlorinated biphenyls analysis. Based on the chromatographic relative retentions of PCDD/F, a software was established for automatic peak recognition of all the isomers from tetra- to octachlorine PCDD/F. It ensured good reliability and accuracy of the analytical data.

Density matrix Loschmidt echo and quantum phase transitions

We introduce the concept of the Loschmidt echo (LE) to the space of the reduced density matrix of spin and fermionic systems to study the density matrix LEs (DMLEs) of the one-dimensional extended Hubbard model and the transverse field Ising model. Our results show that the DMLEs are remarkably influenced by the criticality of the system, and the method is a convenient way to study quantum phase transitions.

The research and progress of micro-fabrication technologies of two-dimensional photonic crystal

The novel material of photonic crystal makes it possible to control a photon, and the photonic integration will have breakthrough progress due to the application of photonic crystal. It is based on the photonic crystal device that the photonic crystal integration could be realized. Therefore, we should first investigate photonic crystal devices based on the active and the passive semiconductor materials, which may have great potential application in photonic integration. The most practical and important method to fabricate two-dimensional photonic crystal is the micro-manufacture method. In this paper, we summarize and evaluate the fabrication methods of two-dimensional photonic crystal in near-infrared region, including electron beam lithography, selection of mask, dry etching, and some works of ours. This will be beneficial to the study of the photonic crystal in China.

Measurement of chirp parameter and modulation index of a semiconductor laser based on optical spectrum analysis

A novel and simple method for measuring the chirp parameter, frequency, and intensity modulation indexes of directly modulated lasers is proposed in a small-signal modulation scheme. A graphical approach is presented. An analytical solution to the measurement of low chirp parameters is also given. The measured results agree well with those obtained using the conventional methods.

A new development on ANN in China - Biomimetic pattern recognition and multi weight vector neurons

A new model of pattern recognition principles-Biomimetic Pattern Recognition, which is based on "matter cognition" instead of "matter classification", has been proposed. As a important means realizing Biomimetic Pattern Recognition, the mathematical model and analyzing method of ANN get breakthrough: a novel all-purpose mathematical model has been advanced, which can simulate all kinds of neuron architecture, including RBF and BP models. As the same time this model has been realized using hardware; the high-dimension space geometry method, a new means to analyzing ANN, has been researched.

A novel contactless method for characterization of semiconductors: Surface electron beam induced voltage in scanning electron microscopy

We present a novel contactless and nondestructive method called the surface electron beam induced voltage (SEBIV) method for characterizing semiconductor materials and devices. The SEBIV method is based on the detection of the surface potential induced by electron beams of scanning electron microscopy (SEM). The core part of the SEBIV detection set-up is a circular metal detector placed above the sample surface. The capacitance between the circular detector and whole surface of the sample is estimated to be about 0.64 pf It is large enough for the detection of the induced surface potential. The irradiation mode of electron beam (e-beam) influences the signal generation. When the e-beam irradiates on the surface of semiconductors continuously, a differential signal is obtained. The real distribution of surface potentials can be obtained when a pulsed e-beam with a fixed frequency is used for irradiation and a lock-in amplifier is employed for detection. The polarity of induced potential depends on the structure of potential barriers and surface states of samples. The contrast of SEBIV images in SEM changes with irradiation time and e-beam intensity.

Intrasubband and intersubband transitions in lightly and heavily doped GaAs/AlGa1-xAs multiple quantum wells

We use a polarizer to investigate quantum-well infrared absorption, and report experimental results as follows. The intrasubband transition was observed in GaAs/AlxGa1-xAs multiple quantum wells (MQWs) when the incident infrared radiation (IR) is polarized parallel to the MQW plane. According to the selection rule, an intrasubband transition is forbidden. Up to now, most studies have only observed the intersubband transition between two states with opposite parity. However, our experiment shows not only the intersubband transitions, but also the intrasubband transitions. In our study, we also found that for light doping in the well (4x10(18) cm(-3)), the intrasubband transition occurs only in the lowest subband, while for the heavy doping (8x10(18) cm(-3)), such a transition occurs not only in the lowest subband, but also in the first excited one, because of the electron subband filling. Further experimental results show a linear dependence of the intrasubband transition frequency on the root of the well doping density. These data are in good agreement with our numerical results. Thus we strongly suggest that such a transition can be attributed to plasma oscillation. Conversely, when the incident IR is polarized perpendicular to the MQW plane, intersubband-transition-induced signals appear, while the intrasubband-transition-induced spectra disappear for both light and heavy well dopings. A depolarization blueshift was also taken into account to evaluate the intersubband transition spectra at different well dopings. Furthermore, we performed a deep-level transient spectroscopy (DLTS) measurement to determine the subband energies at different well dopings. A good agreement between DLTS, infrared absorption, and numerical calculation was obtained. In our experiment, two important phenomena are noteworthy: (1) The polarized absorbance is one order of magnitude higher than the unpolarized spectra. This puzzling result is well explained in detail. (2) When the IR, polarized perpendicular to the well plane, normally irradiates the 45 degrees-beveled edge of the samples, we only observed intersubband transition spectra. However, the intrasubband transition signals caused by the in-plane electric-field component are significantly absent. The reason is that such in-plane electric-field components can cancel each other out everywhere during the light propagating in the samples. The spectral widths of bound-to-bound and bound-to-continuum transitions were also discussed, and quantitatively compared to the relaxation time tau, which is deduced from the electron mobility. The relaxation times deduced from spectral widths of bound-to-bound and bound-to-continuum transitions are also discussed, and quantitatively compared to the relaxation time deduced from electron mobility. [S0163-1829(98)01912-2].

instability of standing wave, global existence and blowup for the klein-gordon-zakharov system with different-degree nonlinearities

This paper discusses the Klein–Gordon–Zakharov system with different-degree nonlinearities in two and three space dimensions. Firstly, we prove the existence of standing wave with ground state by applying an intricate variational argument. Next, by introducing an auxiliary functional and an equivalent minimization problem, we obtain two invariant manifolds under the solution flow generated by the Cauchy problem to the aforementioned Klein–Gordon–Zakharov system. Furthermore, by constructing a type of constrained variational problem, utilizing the above two invariant manifolds as well as applying potential well argument and concavity method, we derive a sharp threshold for global existence and blowup. Then, combining the above results, we obtain two conclusions of how small the initial data are for the solution to exist globally by using dilation transformation. Finally, we prove a modified instability of standing wave to the system under study.

Establishment and characterization of dual-species biofilms formed between a 3,5-dinitrobenzoic-degrading strain and bacteria with high biofilm-forming capabilities

In this study, the possibility of establishing a dual-species biofilm from a bacterium with a high biofilm-forming capability and a 3,5-dinitrobenzoic acid (3,5-DNBA)-degrading bacterium, Comamonas testosteroni A3, was investigated. Our results showed that the combinations of strain A3 with each of five strains with a high biofilm-forming capability (Pseudomonas sp. M8, Pseudomonas putida M9, Bacillus cereus M19, Pseudomonas plecoglossicida M21 and Aeromonas hydrophila M22) presented different levels of enhancement regarding biofilm-forming capability. Among these culture combinations, the 24-h dual-species biofilms established by C. testosteroni A3 with P. putida M9 and A. hydrophila M22 showed the strongest resistance to 3,5-DNBA shock loading, as demonstrated by six successive replacements with DMM2 synthetic wastewater. The degradation rates of 3,5-DNBA by these two culture combinations reached 63.3-91.6% and 70.7-89.4%, respectively, within 6 h of every replacement. Using the gfp-tagged strain M22 and confocal laser scanning microscopy, the immobilization of A3 cells in the dual-species biofilm was confirmed. We thus demonstrated that, during wastewater treatment processes, it is possible to immobilize degrader bacteria with bacteria with a high biofilm-forming capability and to enable them to develop into the mixed microbial flora. This may be a simple and economical method that represents a novel strategy for effective bioaugmentation.

Growth of GaN layers on GaAs and GaP (111) and (001) substrates by molecular beam epitaxy

Films of GaN have been grown using a modified MBE technique in which the active nitrogen is supplied from an RF plasma source. Wurtzite films grown on (001) oriented GaAs substrates show highly defective, ordered polycrystalline growth with a columnar structure, the (0001) planes of the layers being parallel to the (001) planes of the GaAs substrate. Films grown using a coincident As flux, however, have a single crystal zinc-blende growth mode. They have better structural and optical properties. To improve the properties of the wurtzite films we have studied the growth of such films on (111) oriented GaAs and GaP substrates. The improved structural properties of such films, assessed using X-ray and TEM method, correlate with better low-temperature FL.

A voltage-controlled tunable two-color infrared photodetector using GaAs/AlAs/GaAlAs and GaAs/GaAlAs stacked multiquantum wells

A voltage-controlled tunable two-color infrared detector with photovoltaic (PV) and photoconductive (PC) dual-mode operation at 3-5 mu m and 8-14 mu m using GaAs/AlAs/AlGaAs double barrier quantum wells (DBQWs) and bound-to-continuum GaAs/AlGaAs quantum wells is demonstrated. The photoresponse peak of the photovoltaic GaAs/AlAs/GaAlAs DBQWs is at 5.3 mu m, and that of the photoconductive GaAs/GaAlAs quantum wells is at 9.0 mu m. When the two-color detector is under a zero bias, the spectral response at 5.3 mu m is close to saturate and the peak detectivity at 80 K can reach 1.0X10(11) cmHz(1/2)/W, while the spectral photoresponsivity at 9.0 mu m is absolutely zero completely. When the external voltage of the two-color detector is changed to 2.0 V, the spectral photoresponsivity at 5.3 mu m becomes zero while the spectral photoresponsivity at 9.0 mu m increases comparable to that at 5.3 mu m under zero bias, and the peak detectivity (9.0 mu m) at 80 K can reach 1.5X10(10) cmHz(1/2)/W. Strictly speaking, this is a real bias-controlled tunable two-color infrared photodetector. We have proposed a model based on the PV and PC dual-mode operation of stacked two-color QWIPs and the effects of tunneling resonance with narrow energy width of photoexcited electrons in DBQWs, which can explain qualitatively the voltage-controlled tunable behavior of the photoresponse of the two-color infrared photodetector. (C) 1996 American Institute of Physics.