Germ-line transmission of transgenes in Xenopus laevis

Adult Xenopus laevis frogs made transgenic by restriction enzyme-mediated integration were bred to test the feasibility of establishing lines of frogs that express transgenes. All of the 19 animals raised to sexual maturity generated progeny that expressed the transgene(s). The patterns and levels of expression of green fluorescent protein transgenes driven by a viral promoter, rat promoter, and four X. laevis promoters were all unaffected by passage through the germ line. These results demonstrate the ease of establishing transgenic lines in X. laevis.

Highly efficient germ-line transmission of proviral insertions in zebrafish.

An important technology in model organisms is the ability to make transgenic animals. In the past, transgenic technology in zebrafish has been limited by the relatively low efficiency with which transgenes could be generated using either DNA microinjection or retroviral infection. Previous efforts to generate transgenic zebrafish with retroviral vectors used a pseudotyped virus with a genome based on the Moloney murine leukemia virus and the envelope protein of the vesicular stomatitis virus. This virus was injected into blastula-stage zebrafish, and 16% of the injected embryos transmitted proviral insertions to their offspring, with most founders transmitting a single insertion to approximately 2% of their progeny. In an effort to improve this transgenic frequency, we have generated pseudotyped viral stocks of two new Moloney-based genomes. These viral stocks have titers up to two orders of magnitude higher than that used previously. Injection of these viruses resulted in a dramatic increase in transgenic efficiency; over three different experiments, 83% (110/133) of the injected embryos transmitted proviral insertions to 24% of their offspring. Furthermore, founders made with one of the viruses transmitted an average of 11 different insertions through their germ line. These results represent a 50- to 100-fold improvement in the efficiency of generating transgenic zebrafish, making it now feasible for a single lab to rapidly generate tens to hundreds of thousands of transgenes. Consequently, large-scale insertional mutagenesis strategies, previously limited to invertebrates, may now be possible in a vertebrate.

An alternative modal representation of a symmetrical nontransposed three-phase transmission line

The objective of this letter is to propose an alternative modal representation of a nontransposed three-phase transmission line with a vertical symmetry plane by using two transformation matrices. Initially, Clarke's matrix is used to separate the line into components a, 0, and zero. Because a and zero components are not exact modes, they can be considered as being a two-phase line that will be decomposed in its exact modes by using a 2 x 2 modal transformation matrix. This letter will describe the characteristics of the two-phase line before mentioned. This modal representation is applied to decouple a nontransposed three-phase transmission line with a vertical symmetry plane whose nominal voltage is 440 kV.

A new procedure to derive transmission-line parameters: Applications and restrictions

The objective of this paper is to show an alternative methodology to calculate transmission-line parameters per unit length. With this methodology, the transmission-line parameters can be obtained starting from impedances measured in one terminal of the line. First, the article shows the classical methodology to calculate frequency-dependent transmission-line parameters by using Carson's and Pollaczeck's equations for representing the ground effect and Bessel's functions to represent the skin effect. After that, a new procedure is shown to calculate frequency-dependent transmission-line parameters directly from currents and voltages of an existing line. Then, this procedure is applied in a two-phase and a three-phase transmission line whose parameters have been previously calculated by using the classical methodology. Finally, the results obtained by using the new procedure and by using the classical methodology are compared. The article shows simulations results for a typical frequency spectrum of switching transients (10 Hz to 10 kHz).

A new procedure to derive transmission line parameters: Theoretical considerations

The objective of this paper is to show an alternative methodology to calculate transmission line parameters per unit length. With this methodology the transmission line parameters can be obtained starting from the phase currents and voltages in one terminal of the line. First, the article shows the classical methodology to calculate frequency dependent transmission line parameters by using Carson's and Pollaczeck's equations for representing the ground effect and Bessel's functions to represent the skin effect. After that, it is shown a new procedure to calculate frequency dependent transmission line parameters directly from currents and voltages of the line that is already built. Then, this procedure is applied in a two-phase transmission line whose parameters have been previously calculated by using the classical methodology. Finally, the results obtained by using the new procedure and by using the classical methodology are compared. ©2005 IEEE.

Some considerations about a new procedure to derive transmission line parameters concerning three-phase lines

The paper shows an alternative methodology to calculate transmission line parameters per unit length and to apply it in a three-phase line with a vertical symmetry plane. This procedure is derived from a general procedure where the modal transformation matrix of the line is required. In this paper, the unknown modal transformation matrix requested by general procedure is substituted by Clarke's matrix. With the substitution that is shown in the paper, the transmission line parameters can be obtained starting from impedances measured in one terminal of the line. First, the article shows the classical methodology to calculate frequency dependent transmission line parameters by using Carson and Pollaczeck's equations for representing the ground effect and Bessel's functions to represent the skin effect. After that, a new procedure is shown to calculate frequency dependent transmission line parameters directly from currents and voltages of an existing line. Then, this procedure is applied in a non-transposed three-phase transmission line whose parameters have been previously calculated by using the classical methodology. Finally, the results obtained by using the new procedure and by using the classical methodology are compared. The article shows simulation results for typical frequency spectra of switching transients (10 Hz to 10 kHz). Results have shown that procedure has © 2006 IEEE.

Real-time estimation of transmission line impedance based on modal analysis theory

The objective of this paper is to show a methodology to estimate the longitudinal parameters of transmission lines. The method is based on the modal analysis theory and developed from the currents and voltages measured at the sending and receiving ends of the line. Another proposal is to estimate the line impedance in function of the real-time load apparent power and power factor. The procedure is applied for a non-transposed 440 kV three-phase line. © 2011 IEEE.

Eigenvalue analyses for non-transposed three-phase transmission line considering non-implicit ground wires

This paper presents a method for analyzing electromagnetic transients using real transformation matrices in three-phase systems considering the presence of ground wires. So, for the Z and Y matrices that represent the transmission line, the characteristics of ground wires are not implied in the values related to the phases. A first approach uses a real transformation matrix for the entire frequency range considered in this case. This transformation matrix is an approximation to the exact transformation matrix. For those elements related to the phases of the considered system, the transformation matrix is composed of the elements of Clarke's matrix. In part related to the ground wires, the elements of the transformation matrix must establish a relationship with the elements of the phases considering the establishment of a single homopolar reference in the mode domain. In the case of three-phase lines with the presence of two ground wires, it is unable to get the full diagonalization of the matrices Z and Y in the mode domain. This leads to the second proposal for the composition of real transformation matrix: obtain such transformation matrix from the multiplication of two real and constant matrices. In this case, the inclusion of a second matrix had the objective to minimize errors from the first proposal for the composition of the transformation matrix mentioned. © 2012 IEEE.

Sensitivity analysis for overload-relief in transmission lines using an implicitly coupled method (CRIC)

The restructuring of energy markets to provide free access to the networks and the consequent increase of the number of power transactions has been causing congestions in transmission systems. As consequence, the networks suffer overloads in a more frequent way. One parameter that has strong influence on transfer capability is the reactive power flow. A sensitivity analysis can be used to find the best solution to minimize the reactive power flows and relief, the overload in one transmission line. The proposed methodology consists on the computation of two sensitivities based on the use of the Lc matrix from CRIC (Constant Reactive Implicitly Coupled) power flow method, that provide a set of actions to reduce the reactive power flow and alleviate overloads in the lines: (a) sensitivity between reactive power flow in lines and reactive power injections in the buses, (b) sensitivity between reactive power flow in lines and transformer's taps. © 2006 IEEE.

The use of current transformers for noise detection due to sparking in insulators strings in high voltage transmission lines

In a general way, in an electric power utility the current transformers (CT) are used to measurement and protection of transmission lines (TL) 1 The Power Line Carriers systems (PLC) are used for communication between electrical substations and transmission line protection. However, with the increasing use of optical fiber to communication (due mainly to its high data transmission rate and low signal-noise relation) this application loses potentiality. Therefore, other functions must be defined to equipments that are still in using, one of them is detecting faults (short-circuits) and transmission lines insulator strings damages 2. The purpose of this paper is to verify the possibility of using the path to the ground offered by the CTs instead of capacitive couplings / capacitive potential transformers to detect damaged insulators, since the current transformers are always present in all transmission lines (TL's) bays. To this a comparison between this new proposal and the PLC previous proposed system 2 is shown, evaluating the economical and technical points of view. ©2010 IEEE.

WW6: an embryonic stem cell line with an inert genetic marker that can be traced in chimeras.

Mutant mice produced by gene targeting in embryonic stem (ES) cells often have a complex or embryonic lethal phenotype. In these cases, it would be helpful to identify tissues and cell types first affected in mutant embryos by following the contribution to chimeras of ES cells homozygous for the mutant allele. Although a number of strategies for following ES cell development in vivo have been reported, each has limitations that preclude its general application. In this paper, we describe ES cell lines that can be tracked to every nucleated cell type in chimeras at all developmental stages. These lines were derived from blastocysts of mice that carry an 11-Mb beta-globin transgene on chromosome 3. The transgene is readily detected by DNA in situ hybridization, providing an inert, nuclear-localized marker whose presence is not affected by transcriptional or translational controls. The "WW" series of ES lines possess the essential features of previously described ES lines, including giving rise to a preponderance of male chimeras, all of which have to date exhibited germ-line transmission. In addition, clones selected for single or double targeting events form strong chimeras, demonstrating the feasibility of using WW6 cells to identify phenotypes associated with the creation of a null mutant.

Light scattering in polycrystalline alumina with bi-dimensionally large surface grains

Alumina ceramics with high in-line transmittance at 0.5-1.0 mm-thickness were prepared with different doping additives by sintering at 1850 degrees C in vacuum for 1-8 h. Depending on the additive contents and sintering variables bi-dimensionally large surface grains, caused by surface evaporation of MgO, had grown parallel to the surface with similar to 100 mu m thickness and lateral sizes up to the millimeter range. The abnormal grain-growth process also resulted in the formation of pores entrapped inside the large surface grains within a narrow zone at 10-20 mu m distance from the surface. The fraction of these pores is thickness-invariant. Scattering factors associated to the pores entrapped inside the bi-dimensionally large surface grains, second-phase particles, grain-boundaries, and microstructural surface defects are derived from the results of in-line transmission (at 600 nm) and are used together with microstructural characteristics to explain the light transmittance in these materials. (C) 2008 Elsevier Ltd. All rights reserved.

Evolution and pathology in Chagas disease: a review

Trypanosoma cruzi acute infections often go unperceived, but one third of chronically infected individuals die of Chagas disease, showing diverse manifestations affecting the heart, intestines, and nervous systems. A common denominator of pathology in Chagas disease is the minimal rejection unit, whereby parasite-free target host cells are destroyed by immune system mononuclear effectors cells infiltrates. Another key feature stemming from T. cruzi infection is the integration of kDNA minicircles into the vertebrate host genome; horizontal transfer of the parasite DNA can undergo vertical transmission to the progeny of mammals and birds. kDNA integration-induced mutations can enter multiple loci in diverse chromosomes, generating new genes, pseudo genes and knock-outs, and resulting in genomic shuffling and remodeling over time. As a result of the juxtaposition of kDNA insertions with host open reading frames, novel chimeric products may be generated. Germ line transmission of kDNA-mutations determined the appearance of lesions in birds that are indistinguishable from those seen in Chagas disease patients. The production of tissue lesions showing typical minimal rejection units in birds' refractory to T. cruzi infection is consistent with the hypothesis that autoimmunity, likely triggered by integration-induced phenotypic alterations, plays a major role in the pathogenesis of Chagas disease.

Genotypic features of lentivirus transgenic mice.

Lentivector-mediated transgenesis is increasingly used, whether for basic studies as an alternative to pronuclear injection of naked DNA or to test candidate gene therapy vectors. In an effort to characterize the genetic features of this approach, we first measured the frequency of germ line transmission of individual proviruses established by infection of fertilized mouse oocytes. Seventy integrants from 11 founder (G0) mice were passed to 111 first generation (G1) pups, for a total of 255 events corresponding to an average rate of transmission of 44%. This implies that integration had most often occurred at the one- or two-cell stage and that the degree of genotypic mosaicism in G0 mice obtained through this approach is generally minimal. Transmission analysis of eight individual proviruses in 13 G2 mice obtained by a G0-G1 cross revealed only 8% of proviral homozygosity, significantly below the 25% expected from purely Mendelian transmission, suggesting counter-selection due to interference with the functions of targeted loci. Mapping of 239 proviral integration sites in 49 founder animals revealed that about 60% resided within annotated genes, with a marked tendency for clustering in the middle of the transcribed region, and that integration was not influenced by the transcriptional orientation. Transcript levels of a set of arbitrarily chosen target genes were significantly higher in two-cell embryos than in embryonic stem cells or adult somatic cells, suggesting that, as previously noted in other settings, lentiviral vectors integrate preferentially into regions of the genome that are transcriptionally active or poised for activation.

Design and Analysis of Microstrip Lines with EBG-Backed Ground Planes of Different Geometrical Shapes

Propagation of electromagnetic waves through a microstrip line with 2D electromagnetic baud gap (EBG) structures of different geometrical shapes in the ground plane is investigated in this paper. Using transmission-line theory, the design equations for EBG structures are calculated. The measured, numerical. and simulated results are in gone) agreement