Delivery of human apolipoprotein (apo) E to liver by an [E1(-), E3(-), polymerase(-), pTP(-)] adenovirus vector containing a liver-specific promoter inhibits atherogenesis in immunocompetent apoE-deficient mice

Recombinant adenovirus (rAd)-mediated apoE gene transfer to the liver of apoE(-/-) mice is anti-atherogenic. However, first generation rAd vectors were associated with immune clearance of transduced hepatocytes, while an improved [E1(-), E3(-) polymerase(-)] adenovirus vector that persisted in the liver, had transient effects due to cellular shutdown of the cytomegalovirus (CMV) promoter (Ad-CMV-apoE). Here, we utilise an improved class of rAd vector with multiple deletions in the E1, E3, polymerase and pTP (pre-terminal protein) genes, which contains a modular synthetic liver-specific promoter (LSP) to drive expression of the human apoE cDNA (Ad-LSP-apoE) for hepatic gene transfer. Approximately 1 year old apoE(-/-) mice were injected intravenously with 4x10(10) virus particles of either Ad-LSP-apoE or Ad-CMV-apoE. Animals were monitored for plasma apoE, total plasma cholesterol and plasma lipoprotein distribution. The effect of Ad-LSP-apoE on atheroma progression was assessed in animals killed at 8 and 28 weeks after the injections. Ad-LSP-apoE vector administration gave sustained, though low, levels of plasma apoE throughout the study period without inducing a humoral immune response, but failed to reduce plasma cholesterol or normalize the adverse lipoprotein profile. Animals killed 8 weeks after the injections, demonstrated no significant retardation of atherosclerosis, whereas aortic lesions in those killed at 28 weeks were significantly reduced by 30% ( P< 0.006) compared to untreated animals. In summary, the combination of a multiply deleted rAd vector with a liver-specific promoter provided sustained low levels of plasma apoE, resulting in significant retardation of aortic atherosclerotic lesions

Shaping plasmon beams via the controlled illumination of finite-size plasmonic crystals

Plasmonic crystals provide many passive and active optical functionalities, including enhanced sensing, optical nonlinearities, light extraction from LEDs and coupling to and from subwavelength waveguides. Here we study, both experimentally and numerically, the coherent control of SPP beam excitation in finite size plasmonic crystals under focussed illumination. The correct combination of the illuminating spot size, its position relative to the plasmonic crystal, wavelength and polarisation enables the efficient shaping and directionality of SPP beam launching. We show that under strongly focussed illumination, the illuminated part of the crystal acts as an antenna, launching surface plasmon waves which are subsequently filtered by the surrounding periodic lattice. Changing the illumination conditions provides rich opportunities to engineer the SPP emission pattern. This offers an alternative technique to actively modulate and control plasmonic signals, either via micro- and nano-electromechanical switches or with electro- and all-optical beam steering which have direct implications for the development of new integrated nanophotonic devices, such as plasmonic couplers and switches and on-chip signal demultiplexing. This approach can be generalised to all kinds of surface waves, either for the coupling and discrimination of light in planar dielectric waveguides or the generation and control of non-diffractive SPP beams

Handbook of methods for the analysis of the various parameters of the carbon dioxide system in sea water. Version 2

The collection of extensive, reliable, oceanic carbon data is a key component of the Joint Global Ocean Flux Study (JGOFS). A portion of the US JGOFS oceanic carbon dioxide measurements will be made during the World Ocean Circulation Experiment Hydrographic Program. A science team has been formed to plan and coordinate the various activities needed to produce high quality oceanic carbon dioxide measurements under this program. This handbook was prepared at the request of, and with the active participation of, that science team. The procedures have been agreed on by the members of the science team and describe well tested methods. They are intended to provide standard operating procedures, together with an appropriate quality control plan, for measurements made as part of this survey. These are not the only measurement techniques in use for the parameters of the oceanic carbon system; however, they do represent the current state-of-the-art for ship-board measurements. In the end, the editors hope that this handbook can serve widely as a clear and unambiguous guide to other investigators who are setting up to analyze the various parameters of the carbon dioxide system in sea water

Incremental, Inductive Coverability

We give an incremental, inductive (IC3) procedure to check coverability of well-structured transition systems. Our procedure generalizes the IC3 procedure for safety verification that has been successfully applied in finite-state hardware verification to infinite-state well-structured transition systems. We show that our procedure is sound, complete, and terminating for downward-finite well-structured transition systems---where each state has a finite number of states below it---a class that contains extensions of Petri nets, broadcast protocols, and lossy channel systems. We have implemented our algorithm for checking coverability of Petri nets. We describe how the algorithm can be efficiently implemented without the use of SMT solvers. Our experiments on standard Petri net benchmarks show that IC3 is competitive with state-of-the-art implementations for coverability based on symbolic backward analysis or expand-enlarge-and-check algorithms both in time taken and space usage.Comment: Non-reviewed version, original version submitted to CAV 2013; this is a revised version, containing more experimental results and some correction

Retardation of atherosclerosis in immunocompetent apolipoprotein (apo) E-deficient mice followingliver-directed administration of a [E1-, E3-,polymerase-] adenovirus vector containing the elongation factor-1a promoter driving expression of human apoE cDNA

Although gene transfer of human apolipoprotein E (apoE), a 34-kDa circulating glycoprotein, to the liver of apoEdeficient(apoE-/-) mice using recombinant adenoviral vectors (rAd) is antiatherogenic, its full therapeutic potentialhas yet to be realized. First generation vectors led to immune clearance of transduced hepatocytes, while animproved vector with adenovirus regions E1, E3 and DNA polymerase deleted also had transient effects due tocellular shutdown of the cytomegalovirus (CMV) promoter. Here, we have studied an alternative promoter from thecellular elongation factor 1a (EF-1a) gene, injecting 6-8 week old apoE-/- mice intravenously with 2x1010 virusparticles (vp) of the [E1-, E3-, polymerase-] rAd vector Ad-EF1·-apoE. Plasma apoE levels were low (18-55 ng/ml)and failed to reduce plasma cholesterol or normalize the adverse lipoprotein profile. By contrast, thehyperlipidaemic phenotype of apoE-/- mice treated with Ad-CMV-apoE (2x1010 vp) was transiently normalized.Nevertheless, at termination (265 days) the aortic lesion areas in animals given Ad-EF1·-apoE were significantlyreduced by 15% (P<0.05) compared to untreated animals, a decrease approaching that in Ad-CMV-apoE-treatedmice (23%; P<0.02). Importantly, the attenuation of apoE transgene expression noted with the CMV promoter wasabsent with the EF-1a promoter, which gave relatively sustained, albeit low, levels of plasma apoE throughout thestudy period

Self-consistent symmetries in the proton-neutron Hartree-Fock-Bogoliubov approach

Symmetry properties of densities and mean fields appearing in the nuclear Density Functional Theory with pairing are studied. We consider energy functionals that depend only on local densities and their derivatives. The most important self-consistent symmetries are discussed: spherical, axial, space-inversion, and mirror symmetries. In each case, the consequences of breaking or conserving the time-reversal and/or proton-neutron symmetries are discussed and summarized in a tabulated form, useful in practical applications.Comment: 26 RevTex pages, 1 eps figure, 9 tables, submitted to Physical Review

Functional rescue of dystrophin deficiency in mice caused by frameshift mutations using Campylobacter jejuni Cas9

Duchenne muscular dystrophy (DMD) is a fatal, X-linked muscle wasting disease caused by mutations in the DMD gene. In 51% of DMD cases, a reading frame is disrupted because of deletion of several exons. Here, we show that CjCas9 derived from Campylobacter jejuni can be used as a gene editing tool to correct an out-of-frame Dmd exon in Dmd knockout mice. Herein, we used Cas9 derived from S. pyogenes to generate Dmd knockout (KO) mice with a frameshift mutation in Dmd gene. Then, we expressed CjCas9, its single-guide RNA, and the eGFP gene in the tibialis anterior muscle of the Dmd KO mice using an all-in-one adeno-associated virus (AAV) vector. CjCas9 cleaved the target site in the Dmd gene efficiently in vivo and induced small insertions or deletions at the target site. This treatment resulted in conversion of the disrupted Dmd reading frame from out-of-frame to in-frame, leading to the expression of dystrophin in the sarcolemma. Importantly, muscle strength was enhanced in the CjCas9-treated muscles, without off-target mutations, indicating high efficiency and specificity of CjCas9. This work suggests that in vivo DMD frame correction, mediated by CjCas9 has great potential for the treatment of DMD and other neuromuscular diseases

Mixed-mode oscillations and interspike interval statistics in the stochastic FitzHugh-Nagumo model

We study the stochastic FitzHugh-Nagumo equations, modelling the dynamics of neuronal action potentials, in parameter regimes characterised by mixed-mode oscillations. The interspike time interval is related to the random number of small-amplitude oscillations separating consecutive spikes. We prove that this number has an asymptotically geometric distribution, whose parameter is related to the principal eigenvalue of a substochastic Markov chain. We provide rigorous bounds on this eigenvalue in the small-noise regime, and derive an approximation of its dependence on the system's parameters for a large range of noise intensities. This yields a precise description of the probability distribution of observed mixed-mode patterns and interspike intervals.Comment: 36 page

Elliptic Solitons and Groebner Bases

We consider the solution of spectral problems with elliptic coefficients in the framework of the Hermite ansatz. We show that the search for exactly solvable potentials and their spectral characteristics is reduced to a system of polynomial equations solvable by the Gr\"obner bases method and others. New integrable potentials and corresponding solutions of the Sawada-Kotera, Kaup-Kupershmidt, Boussinesq equations and others are found.Comment: 18 pages, no figures, LaTeX'2

Recommendations for the Determination of Nutrients in Seawater to High Levels of Precision and Inter-Comparability using Continuous Flow Analysers

The Global Ocean Ship-based Hydrographic Investigations Program (GO-SHIP) brings together scientists with interests in physical oceanography, the carbon cycle, marine biogeochemistry and ecosystems, and other users and collectors of ocean interior data to develop a sustained global network of hydrographic sections as part of the Global Ocean Climate Observing System. A series of manuals and guidelines are being produced by GO-SHIP which update those developed by the World Ocean Circulation Experiment (WOCE) in the early 1990s. Analysis of the data collected in WOCE suggests that improvements are needed in the collection of nutrient data if they are to be used for determining change within the ocean interior. Production of this manual is timely as it coincides with the development of reference materials for nutrients in seawater (RMNS). These RMNS solutions will be produced in sufficient quantities and be of sufficient quality that they will provide a basis for improving the consistency of nutrient measurements both within and between cruises. This manual is a guide to suggested best practice in performing nutrient measurements at sea. It provides a detailed set of advice on laboratory practice for all the procedures surrounding the use of 1 gas-segmented continuous flow analysers (CFA) for the determination of dissolved nutrients (usually ammonium, nitrate, nitrite, phosphate and silicate) at sea. It does not proscribe the use of a particular instrument or related chemical method as these are well described in other publications. The manual provides a brief introduction to the CFA method, the collection and storage of samples, considerations in the preparation of reagents and the calibrations of the system. It discusses how RMNS solutions can be used to “track” the performance of a system during a cruise and between cruises. It provides a format for the meta-data that need to be reported along side the sample data at the end of a cruise so that the quality of the reported data can be evaluated and set in context relative to other data sets. Most importantly the central manual is accompanied by a set of nutrient standard operating procedures (NSOPs) that provide detailed information on key procedures that are necessary if best quality data are to be achieved consistently. These cover sample collection and storage, an example NSOP for the use of a CFA system at sea, high precision preparation of calibration solutions, assessment of the true calibration blank, checking the linearity of a calibration and the use of internal and externally prepared reference solutions for controlling the precision of data during a cruise and between cruises. An example meta-data report and advice on the assembly of the quality control and statistical data that should form part of the meta-data report are also given