De novo generation of a non-segmented negative strand RNA virus with a bicistronic gene

Reverse genetics has facilitated the use of non-segmented negative strand RNA viruses (NNSV) as vectors. Currently, heterologous gene expression necessitates insertion of extra-numeral transcription units (ENTUs), which may alter the NNSV polar transcription gradient and attenuate growth relative to wildtype (Wt). We hypothesized that rescuing recombinant Sendai Virus (rSeV) with a bicistronic gene might circumvent this attenuation but still allow heterologous open reading frame (ORF) expression. Therefore, we used a 9-nucleotide sequence previously described with internal ribosome entry site (IRES) activity, which, when constructed as several repeats, synergistically increased the level of expression of the second cistron [Chappell, S.A., Edelman, G.M., Mauro, V.P., 2000. A 9-nt segment of a cellular mRNA can function as an internal ribosome entry site (IRES) and when present in linked multiple copies greatly enhances IRES activity. Proc. Natl. Acad. Sci. U.S.A. 97, 1536-1541]. We inserted the Renilla luciferase (rLuc) ORF, preceded by 1, 3 or 7 IRES copies, downstream of the SeV N ORF in an infectious clone. Corresponding rSeVs were successfully rescued. Interestingly, bicistronic rSeVs grew as fast as or faster than Wt rSeV. Furthermore, SeV gene transcription downstream of the N/rLuc gene was either equivalent to, or slightly enhanced, compared to Wt rSeV. Importantly, all rSeV/rLuc viruses efficiently expressed rLuc. IRES repetition increased rLuc expression at a multiplicity of infection of 0.1, although without evidence of synergistic enhancement. In conclusion, our approach provides a novel way of insertion and expression of foreign genes in NNSVs. (C) 2008 Elsevier B.V. All rights reserved.

A Low Complexity Video Watermarking in H.264 Compressed Domain

In this paper, a new blind and readable H.264 compressed domain watermarking scheme is proposed in which the embedding/extracting is performed using the syntactic elements of the compressed bit stream. As a result, it is not necessary to fully decode a compressed video stream both in the embedding and extracting processes. The method also presents an inexpensive spatiotemporal analysis that selects the appropriate submacroblocks for embedding, increasing watermark robustness while reducing its impact on visual quality. Meanwhile, the proposed method prevents bit-rate increase and restricts it within an acceptable limit by selecting appropriate quantized residuals for watermark insertion. Regarding watermarking demands such as imperceptibility, bit-rate control, and appropriate level of security, a priority matrix is defined which can be adjusted based on the application requirements. The resulted flexibility expands the usability of the proposed method.

THz Frequency Selective Surface Filters for Earth Observation Remote Sensing Instruments

The purpose of this paper is to review recent developments in the design and fabrication of Frequency Selective Surfaces (FSS) which operate above 300 GHz. These structures act as free space electromagnetic filters and as such provide passive remote sensing instruments with multispectral capability by separating the scene radiation into separate frequency channels. Significant advances in computational electromagnetics, precision micromachining technology and metrology have been employed to create state of the art FSS which enable high sensitivity receivers to detect weak molecular emissions at THz wavelengths. This new class of quasi-optical filter exhibits an insertion loss

Sub-mm wet etched linear to circular polarization fss based polarization converters

This communication investigates the potential for fabrication of micromachined silicon sub-millimeter wave periodic arrays of freestanding slot frequency selective surfaces (FSS) using wet etch KOH technology. The vehicle for this is an FSS for generating circularly polarized signals from an incident linearly polarized signal at normal incidence to the structure. Principal issues and fabrication processes involved from the initial design of the core FSS structures to be made and tested through to their final testing are addressed. Measured and simulated results for crossed and ring slot element shapes in single and double layer polarization convertor structures are presented for sub-mm wave operation. It is shown that 3 dB axial ratio (AR) bandwidths of 21% can be achieved with the one layer perforated screen design and that the rate of change is lower than the double layer structures. An insertion loss of 1.1 dB can be achieved for the split circular ring double layer periodic array. These results are shown to be compatible with the more specialized fabrication equipment dry reactive ion etching approach previously used for the construction of this type of structure. © 2011 IEEE.

Alkynyl-Bridged Ruthenium(II) 4 '-Diferroceny1-2,2 ':6 ',2 ''-terpyridine Electron Transfer Complexes: Synthesis, Structures, and Electrochemical and Spectroscopic Studiese

The novel ligand 4'-diferrocenylallcyne-2,2':6',2 ''-terpyridine (7; Fc-C C-Fc-tpy; tpy = terpyridyl; Fc = ferrocenyl) and its Ru2+ complexes 8-10 have been synthesized and characterized by single-crystal X-ray diffraction, cyclic voltammetry, and UV-vis and luminescence spectroscopy. Electrochemical data and UV absorption and emission spectra indicate that the insertion of an ethynyl group causes delocalization of electrons in the extended pi* orbitals. Cyclic voltammetric measurements of 7 show two successive reversible one-electron-oxidation processes with half-wave potentials of 0.53 and 0.78 V. The small variations of the E-1/2 values for the Fe2+/Fe3+ redox couples after the coordination of the Ru2+ ion suggest a weak interaction between the Ru2+ and Fe2+ centers. After insertion of an ethynyl group, UV-vis absorption spectra show a red shift of the absorption peak of the (1)[(d(pi)(Fe))(6)]->(1)[(d(pi)(Fe))(5)(pi*(Ru)(tpy))(1)] MMLCT of the Ru2+ complexes. The Ru2+ complex 8 exhibits the strongest luminescence intensity (lambda(em)(max) 712 nm, Phi(em) = 2.63 x 10(-4), tau = 323 ns) relative to analogous ferrocene-based terpyridine Ru(II) complexes in H2O/CH3CN (4/1 v/v) solution.

Molecular differences between two Jeryl Lynn mumps virus vaccine component strains, JL5 and JL2

The Jeryl Lynn (JL) vaccine against mumps virus (MuV) contains two components, MuV(JL5) and MuV(JL2), which differ by over 400 nt. Due to the occurrence of bias in the direction of mutation, these differences and those found in nucleotide sequences of different isolates of the minor component in the vaccine (MuV(JL2)) might be due to the effect of ADAR-like deaminases on MuV grown in tissue-cultured cells. A molecular clone Of MuV(JL2) (pMuV(JL2)) and MuV(JL2) -specific helper plasmids were constructed in order to investigate molecular interactions between MuV(JL5) and MuV(JL2), to augment the existing molecular clone Of MuV(JL)5 (pMuV(JL5)) and MuV(JL5) -specific helper plasmids. Genome and mRNA termini Of MuV(JL2) were characterized, and an unusual oligo-G insertion transcriptional editing event was detected near the F mRNA polyadenylation site of MuV(JL2), but not Of MuV(JL5). Genes encoding glycoproteins of rMuV(JL2) and rMuV(JL5) have been exchanged to characterize the oligo-G insertion, which associated with the specific sequence of the IF gene of MuV(JL2) and not with any other genes or the RNA-dependent RNA polymerase of strain MuV(JL2). The results indicate that a single G-to-A sequence change obliterates the co-transcriptional editing of the F mRNA and that this oligo-G insertion does not affect the growth of the virus.

Phase behavior of mixtures of colloidal platelets and nonadsorbing polymers

The phase behavior of a model system of colloidal platelets and nonadsorbing polymers is studied using computer simulations and perturbation theory. The equation of state for the pure platelet reference system is obtained by Monte Carlo simulations, and the free volume fraction accessible to polymers is measured by a trial insertion method. The free volume fraction is also calculated using scaled particle theory. Subsequently, the phase diagram for platelet-polymer mixtures is calculated. For a platelet aspect ratio L/D=0.1 and a polymer to platelet size ratio d/D>0.2, we observe coexistence between two isotropic phases with different densities. For smaller polymers d/D

Determination of the Physical Properties of Room Temperature Ionic Liquids Using a Love Wave Device

In this work, we have shown that a 100 MHz Love wave device can be used to determine whether room temperature ionic liquids (RTILs) are Newtonian fluids and have developed a technique that allows the determination of the density-viscosity product, rho eta of a Newtonian RTIL. In addition, a test for a Newtonian response was established by relating the phase change to insertion loss change. Five concentrations of a water-miscible RTIL and seven pure RTILs were measured. The changes in phase and insertion loss were found to vary linearly with the square root of the density-viscosity product for values up to (rho eta)(1/2) similar to 10 kg m(-2) s(-1/2). The square root of the density-viscosity product was deduced from the changes in either phase or insertion loss using glycerol as a calibration liquid. In both cases, the deduced values of rho eta agree well with those measured using viscosity and density meters. Miniaturization of the device, beyond that achievable with the lower-frequency quartz crystal microbalance approach, to measure smaller volumes is possible. The ability to fabricate Love wave and other surface acoustic wave sensors using planar metallization technologies gives potential for future integration into lab-on-a-chip analytical systems for characterizing ionic liquids.

Ultrafast Low-Loss 40-70 GHz SPST Switch

The design and characterizations of an ultrafast single-pole single-throw (SPST) absorptive differential switch are presented. The switch exhibits low insertion loss less than 1 dB, and isolation better than 16 dB from 40 to 70 GHz. Sub-nanosecond switching time is achieved by adopting a differential current-steering technique. The total measured rise and fall time are 75 ps envisaging that switching rates up to 13 Gb/s are achievable. To our best knowledge, this is the fastest, lowest insertion loss V-band SPST switch yet reported that can operate over a wide bandwidth of 30 GHz.

Frequency selective surface using nested split ring slot elements as a lens with mechanically reconfigurable beam steering capability

The design is described of a double layer frequency selective surface which can produce a differential phase shift of 180 ° as the wave propagates through it at normal incidence. The hand of an applied circularly polarized signal is reversed due to the 180° phase shift, and it is demonstrated that the exit circularly polarized output signal can be phase advanced or phase retarded by 180 ° upon rotation of the elements comprising the structure. This feature allows the surface to act as a spatial phase shifter. In this paper the beam steering capabilities of a 10 × 10 array of such elements are demonstrated. Here the individual elements comprising the array are rotated relative to each other in order to generate a progressive phase shift. At normal incidence the 3 dB Axial Ratio Bandwidth for LHCP to RHCP conversion is 5.3% and the insertion loss was found to be -2.3 dB, with minimum axial ratio of 0.05 dB. This array is shown to be able to steer a beam from -40 ° to +40 ° while holding axial ratio at the pointing angle to below 4 dB. The measured radiation patterns match the theoretical calculation and full-wave simulation results. © 2010 IEEE.

Inline Interdigital Pseudo-Elliptic Helical Resonator Filters

A new inline coupling topology for narrowband helical resonator filters is proposed that allows to introduce selectively located transmission zeros (TZs) in the stopband. We show that a pair of helical resonators arranged in an interdigital configuration can realize a large range of in-band coupling coefficient values and also selectively position a TZ in the stopband. The proposed technique dispenses the need for auxiliary elements, so that the size, complexity, power handling and insertion loss of the filter are not compromised. A second order prototype filter with dimensions of the order of 0.05 lambda, power handling capability up to 90 W, measured insertion loss of 0.18 dB and improved selectivity is presented.

Applied aspects of Rhodococcus genetics

Eubacteria of the genus Rhodococcus are a diverse group of microorganisms commonly found in many environmental niches from soils to seawaters and as plant and animal pathogens. They exhibit a remarkable ability to degrade many organic compounds and their economic importance is becoming increasingly apparent. Although their genetic organisation is still far from understood, there have been many advances in recent years. Reviewed here is the current knowledge of rhodococci relating to gene transfer, recombination, plasmid replication and functions, cloning vectors and reporter genes, gene expression and its control, bacteriophages, insertion sequences and genomic rearrangements. Further fundamental studies of Rhodococcus genetics and the application of genetic techniques to the these bacteria will be needed for their continued biotechnological exploitation.

The Sound of Becoming: Moving Towards a Virtual Music

In 1985 Jacques Attali proposed a new modality for music. He suggests that there be “not a new music, but a new way of making music... a radically new form of the insertion of music into communication” (Attali 134). What Attali foretold has become a firm reality in contemporary musical practice. One has only to look at any current musical activity to encounter work that relies heavily on real-time interaction and dynamic generation and/or modification of materials. But why is this ontologically different ‘mode of essentially interactive and transformative existence’ (Ziarek 195), this ‘new way of making music’, so attractive to contemporary artists? What is motivating artists to abandon a production model in favor of a model of real-time interactive exploration? I will argue that at the foundation of this new artistic ontology lies Deleuze’s concept of the virtual. It is a recognition of the virtual power of music, that music making can be an act of invention, a process where one can discover something never before experienced.

Thermal Deformation Analysis of Micro satellite Structure

Mounting accuracy of satellite payload and ADCS (attitude determination and control subsystem) seats is one of the requirements to achieve the satellite mission with acceptable performance. Components of mounting inaccuracy are technological inaccuracies, residual plastic deformations after loading (during transportation and orbital insertion), elastic deformations, and thermal deformations during orbital operation. This paper focuses on estimation of thermal deformations of satellite structure. Thermal analysis is executed by applying finite-difference method (IDEAS) and temperature profile for satellite components case is evaluated. Then, Perform thermal finite-element analysis applying the finite-difference model results as boundary conditions; and calculate the resultant thermal strain. Next, applying the resultant thermal strain, perform finite-element structure analysis to evaluate structure deformations at the payload and ADCS equipments seats.

Distinct functional domains of the Salmonella enterica WbaP transferase that is involved in the initiation reaction for synthesis of the O antigen subunit

WbaP is a membrane enzyme that initiates O antigen synthesis in Salmonella enterica by catalysing the transfer of galactose 1-phosphate (Gal-1-P) onto undecaprenyl phosphate (Und-P). WbaP possesses at least three predicted structural domains: an N-terminal region containing four transmembrane helices, a large central periplasmic loop, and a C-terminal domain containing the last transmembrane helix and a large cytoplasmic tail. In this work, we investigated the contribution of each region to WbaP function by constructing a series of mutant WbaP proteins and using them to complement O antigen synthesis in DeltawbaP mutants of S. enterica serovars Typhi and Typhimurium. Truncated forms of WbaP lacking the periplasmic loop exhibited altered chain-length distributions in O antigen polymerization, suggesting that this central domain is involved in modulating the chain-length distribution of the O polysaccharide. The N-terminal and periplasmic domains were dispensable for complementation of O antigen synthesis in vivo, suggesting that the C-terminal domain carries the sugar-phosphate transferase activity. However, despite the fact that they complemented the synthesis of O antigen in the DeltawbaP mutant in vivo, membrane extracts containing WbaP derivatives without the N-terminal domain failed to transfer radioactive Gal from UDP-Gal into a lipid-rich fraction. These results suggest that the N-terminal region of WbaP, which contains four transmembrane domains, is essential for the insertion or stability of the protein in the bacterial membrane. We propose that the domain structure of WbaP enables this protein not only to function in the transfer of Gal-1-P to Und-P but also to establish critical interactions with additional proteins required for the correct assembly of O antigen in S. enterica.