995 resultados para Cross-amplification


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This paper investigates the vibration dynamics of a closed-chain, cross-coupled architecture of MEMS resonators. The system presented here is electrostatically transduced and operates at 1.04 MHz. Curve veering of the eigenvalue loci is used to experimentally quantify the coupling spring constants. Numerical simulations of the motional resistance variation against induced perturbation are used to assess the robustness of the cross-coupled system as opposed to equivalent traditional open-ended linear one-dimensional coupling scheme. Results show improvements of as much as 32% in the motional resistance between the cross-coupled system and its one-dimensional counterpart. © 2013 IEEE.

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A novel gene-K23, differentially expressed in cross-subfamily cloned embryos, was isolated by RACE-PCR technique. It had 2580 base pairs (bp) in length, with a 1,425 bp open reading frame (ORF) encoding a putative protein of 474 amino acids (aa). Bioinformatic analysis indicated that K23 had 22 phosphorylation sites, but it had no signal peptides. Developmental expression analysis in zebrafish showed that K23 transcripts were maternally expressed in ovum and the amount of K23 transcripts increased gradually from zygote to pharyngula period. Subcellular localization analysis revealed that K23 protein was homogeneously distributed both in nuclei and cytoplasm. Taken together, our findings indicate that K23 gene is a novel gene differentially expressed in fish cross-subfamily cloned embryos.

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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.

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Cross-species nuclear transfer (NT) has been used to retain the genetic viability of a species near extinction. However, unlike intra-species NT, most embryos produced by cross-species NT were unable to develop to later stages due to incompatible nucleocytoplasmic interactions between the donor nuclei and the recipient cytoplasm from different species. To study the early nucleocytoplasmic interaction in cross-species NT, two laboratory fish species (zebrafish and rare minnow) from different subfamilies were used to generate cross-subfamily NT embryos in the present study. Suppression subtractive hybridization (SSH) was performed to screen out differentially expressed genes from the forward and reverse subtractive cDNA libraries. After dot blot and real-time PCR analysis, 80 of 500 randomly selective sequences were proven to be differentially expressed in the cloned embryos. Among them, 45 sequences shared high homology with 28 zebrafish known genes, and 35 sequences were corresponding to 22 novel expressed sequence tags (ESTs). Based on functional clustering and literature mining analysis, up-and down-regulated genes in the cross-subfamily cloned embryos were mostly relevant to transcription and translation initiation, cell cycle regulation, protein binding, etc. To our knowledge, this is the first report on the determination of genes involved in the early development of cross-species NT embryos of fish. (C) 2007 Elsevier Inc. All rights reserved.

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Expressed sequence tags (ESTs) are a source for microsatellite development. In the present study, EST-derived microsatelltes (EST-SSRs) were generated and characterized in the common carp (Cyprinus carpio) by data mining from updated public EST databases and by subsequent testing for polymorphism. About 5.5% (555) of 10,088 ESTs contain repeat motifs of various types and lengths with CA being the most abundant dinucleotide one. Out of the 60 EST-SSRs for which PCR primers were designed, 25 loci showed polymorphism in a common carp population with the alleles per locus ranging from 3 to 17 (mean 7). The observed (H-O) and expected (HE) heterozygosities of these EST-SSRs were 0.13-1.00 and 0.12-0.91, respectively. Six EST-SSR loci significantly deviated from the Hardy-Weinberg equilibrium (HWE) expectation, and the remaining 19 loci were in HWE. Of the 60 primer sets, the rates of polymorphic EST-SSRs were 42% in common carp, 17% in crucian carp (Carassius auratus), and 5% in silver carp (Hypophthalmichthys molitrix), respectively. These new EST-SSR markers would provide sufficient polymorphism for population genetic studies and genome mapping of the common carp and its closely related fishes. (c) 2007 Published by Elsevier B.V.

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Wuchang bream (Megalobrama amblycephala) is an economically important fish in China. From a (GT)(13)-enriched genomic library, 20 microsatellites were developed. Nine of these 20 loci were polymorphic in a test population with allele numbers ranging from two to four, and the observed and expected heterozygosities ranging from 0.2609 to 0.7826 and from 0.3739 to 0.7546, respectively. In the cross-species amplifications, six of these nine loci were also polymorphic in white amur bream (Parabramis pekinensis). These polymorphic microsatellite loci are potentially useful for population genetics of Wuchang bream and its closely related species.

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A process is presented for the forming of variable cross-section I-beams by hot rolling. Optimized I-beams with variable cross-section offer a significant weight advantage over prismatic beams. By tailoring the cross-section to the bending moment experienced within the beam, around 30% of the material can be saved compared to a standard section. Production of such beams by hot rolling would be advantageous, as It combines high volume capacity with high material yields. Through controlled variation of the roll gap during multiple passes, beams with a variable cross-section have been created using shaped rolls similar to those used for conventional I-beam rolling. The process was tested experimentally on a small scale rolling mill, using plasticine as the modelling material. These results were then compared to finite element simulations of individual stages of the process conducted using Abaqus/Standard. Results here show that the process can successfully form a beam with a variable depth web. The main failure modes of the process, and the limitations on the achievable variations In geometry are also presented. Finally, the question of whether or not optimal beam geometries can be created by this process Is discussed. © 2011 Wiley-VCH Verlag GmbH & Co. KGaA. Weinheim.

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In previous studies of nuclear transplantation, most cloned animals were obtained by intraspecies nuclear transfer and are phenotypically identical to their nuclear donors; furthermore, there was no further report on successful fish cloning since the report of cloned zebrafish. Here we report the production of seven cross-genus cloned fish by transferring nuclei from transgenic common carp into enucleated eggs of goldfish. Nuclear genomes of the cloned fish were exclusively derived from the nuclear donor species, common carp, whereas the mitochondrial DNA from the donor carp gradually disappeared during the development of nuclear transfer (NT) embryos. The somite development process and somite number of nuclear transplants were consistent with the recipient species, goldfish, rather than the nuclear donor species, common carp. This resulted in a long-lasting effect on the vertebral numbers of the cloned fish, which belonged to the range of goldfish. These demonstrate that fish egg cytoplasm not only can support the development driven by transplanted nuclei from a distantly related species at the genus scale but also can modulate development of the nuclear transplants.

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The sustainable remediation concept, aimed at maximizing the net environmental, social, and economic benefits in contaminated site remediation, is being increasingly recognized by industry, governments, and academia. However, there is limited understanding of actual sustainable behaviour being adopted and the determinants of such sustainable behaviour. The present study identified 27 sustainable practices in remediation. An online questionnaire survey was used to rank and compare them in the US (n=112) and the UK (n=54). The study also rated ten promoting factors, nine barriers, and 17 types of stakeholders' influences. Subsequently, factor analysis and general linear models were used to determine the effects of internal characteristics (i.e. country, organizational characteristics, professional role, personal experience and belief) and external forces (i.e. promoting factors, barriers, and stakeholder influences). It was found that US and UK practitioners adopted many sustainable practices to similar extents. Both US and UK practitioners perceived the most effectively adopted sustainable practices to be reducing the risk to site workers, protecting groundwater and surface water, and reducing the risk to the local community. Comparing the two countries, we found that the US adopted innovative in-situ remediation more effectively; while the UK adopted reuse, recycling, and minimizing material usage more effectively. As for the overall determinants of sustainable remediation, the country of origin was found not to be a significant determinant. Instead, organizational policy was found to be the most important internal characteristic. It had a significant positive effect on reducing distant environmental impact, sustainable resource usage, and reducing remediation cost and time (p<0.01). Customer competitive pressure was found to be the most extensively significant external force. In comparison, perceived stakeholder influence, especially that of primary stakeholders (site owner, regulator, and primary consultant), did not appear to have as extensive a correlation with the adoption of sustainability as one would expect.

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The hole-mediated ferromagnetism in (In,Mn)As quantum dots is investigated using the k center dot p method and the mean field model. It is found that the (In,Mn)As quantum dot can be ferromagnetic at room temperature when there is one hole in the dot. For the spherical quantum dots, the Curie temperature decreases as the diameter increases, and increases as the effective composition of magnetic ions increases. It is interesting to find that the (In,Mn)As oblate quantum dot has highly anisotropic Zeeman splitting and ferromagnetism due to the spin-orbit coupling effect, which can be used as an uniaxial spin amplifier. (c) 2008 American Institute of Physics.

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We present a comprehensive numerical study on the all-optical wavelength conversion based on the degenerate four-wave-mixing with continuous-wave pumping in the silicon nanowire waveguide. It is well known that the conversion efficiency and the 3-dB bandwidth can be greatly affected by the phase-matching condition. Through proper design of the waveguide cross-section, its dispersion property can be adjusted to satisfy the phase-matching condition and therefore effective wavelength conversion can be achieved in a large wavelength range. Generally, the group velocity dispersion plays a dominant role in the wavelength conversion. However, the fourth-order dispersion takes an important effect on the wavelength conversion when the group velocity dispersion is near the zero-point. Furthermore, the conversion efficiency and the 3-dB bandwidth can also be affected by the interactive length and the initial pump power. Through the numerical simulation, the optimal values for the interactive length and the initial pump power, which are functions of the propagation loss, are obtained to realize the maximum conversion efficiency. (C) 2008 Elsevier B.V. All rights reserved.

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The single-mode condition for silicon rib waveguides with large cross sections is obtained using a numerical method based on the finite-difference beam propagation method (FD-BPM). An ultrawide computation window is used to contain the wide mode profile near cutoff. Comparison with previous results shows that the formula predicted by the mode-matching technique is in a better agreement with our results. (C) 2004 Society of Photo-Optical Instrumentation Engineers.

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Single-mode condition for silicon rib waveguides with trapezoidal cross-section was obtained using a numerical method based on imaginary-distance beam propagation method with non-uniform discretization. Both quasi-transverse-electric and quasi-transverse-magnetic modes were investigated. Simulated single-mode condition is given by a modified equation. Comparison with reported results shows that the Marcatili's method is in a better agreement with our results. (C) 2003 Elsevier B.V. All rights reserved.

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A 3-dB multimode interference optical coupler based on rib waveguides with trapezoidal cross section was designed and fabricated on silicon-on-insulator wafer. Potassium hydroxide (KOH) anisotropic chemical etching of silicon was used to fabricate the waveguides to obtain smooth interface. A modified finite-difference beam propagation method was used to simulate the multimode rib waveguide with slope interfaces. The rms roughness of etching interface is as small as 1.49 nm. The propagation loss of the waveguide is 1.3 dB/cm at wavelength of 1.55 mum. The fabricated 3-dB coupler has a good uniformity of 0.2 dB.