Supramolecular Assembly of Aminoethylene‐Lipopeptide PMO Conjugates into RNA Splice‐Switching Nanomicelles

Phosphorodiamidate morpholino oligomers (PMOs) are oligonucleotide analogs that can be used for therapeutic modulation of pre‐mRNA splicing. Similar to other classes of nucleic acid‐based therapeutics, PMOs require delivery systems for efficient transport to the intracellular target sites. Here, artificial peptides based on the oligo(ethylenamino) acid succinyl‐tetraethylenpentamine (Stp), hydrophobic modifications, and an azide group are presented, which are used for strain‐promoted azide‐alkyne cycloaddition conjugation with splice‐switching PMOs. By systematically varying the lead structure and formulation, it is determined that the type of contained fatty acid and supramolecular assembly have a critical impact on the delivery efficacy. A compound containing linolenic acid with three cis double bonds exhibits the highest splice‐switching activity and significantly increases functional protein expression in pLuc/705 reporter cells in vitro and after local administration in vivo. Structural and mechanistic studies reveal that the lipopeptide PMO conjugates form nanoparticles, which accelerate cellular uptake and that the content of unsaturated fatty acids enhances endosomal escape. In an in vitro Duchenne muscular dystrophy exon skipping model using H2K‐mdx52 dystrophic skeletal myotubes, the highly potent PMO conjugates mediate significant splice‐switching at very low nanomolar concentrations. The presented aminoethylene‐lipopeptides are thus a promising platform for the generation of PMO‐therapeutics with a favorable activity/toxicity profile.     

Simulating Ramsey-Type Fringes in a Pulsed Microwave-Driven Classical Josephson Junction

We present evidence for a close analogy between the nonlinear behaviour of a pulsed microwave-driven Josephson junction at low temperature and the experimentally observed behaviour of Josephson systems operated below the quantum transition temperature under similar conditions. We specifically address observations of Ramsey-type fringe oscillations, which can be understood in classical nonlinear dynamics as results of slow transient oscillations in a pulsed microwave environment. Simulations are conducted to mimic experimental measurements by recording the statistics of microwave-induced escape events from the anharmonic potential well of a zero-voltage state. Observations consistent with experimentally obtained Ramsey-type oscillations are found in the classical model. An erratum to this article can be found at     

Uncertainty in health risk analysis

This paper discuss several quantitative issues that arise in the analysis of health risks, beginning with principles such as de minimis and zero-risk. The paper also provides a probabilistic definition of risk in terms of hazard, context, consequence, magnitude, and uncertainty. The example relies on this definition to investigate, through sensitivity analysis, the effect that uncertainty has on the results obtained. The results, from a case study based on waterborne total arsenic, show that the choice of dose—response functions causes more uncertainty than any other component of risk analysis. Chemical carcinogenesis provides the basis for discussing inability to know as well as uncertainty. The conclusion is that risk analysis keeps uncertainty and inability to know separate; through this function, it provides a much needed method to present information to decision makers and the public.     

Fabrication and Characterization of YBa2Cu3O7 − x Thin Film Step Junctions on MgO Substrates

We have investigated the tunneling properties of proximity-effect superconductor-normal-superconductor junctions based on high-temperature superconductor YBCO thin films and normal Ag layer. The systematic analysis of the properties of the films and the fabrication process which leads to a good-quality junctions is discussed in detail. The properties of the tunneling barrier are investigated by dc and rf measurements: the dc properties reveal that our junctions fully behave as expected from the de Gennes model. Regular Shapiro steps in the current–voltage characteristics are obtained as a result of microwave irradiation, indicating reliable rf properties of the junctions.     

A Model of Weekly Labor Participation for a Belgian Synthetic Population

This paper presents a model of labor participation calibrated on a weekly basis; part-time and full-time employments are also considered. By applying the theory of random utility maximization we model households’ choices. In order to have a good temporal and spatial coverage the model is calibrated on three datasets, extracted from both national and regional travel surveys. The results are applied to synthetic households that reproduce the Belgian population. The proposed innovative methodology simulates synthetic agents by accounting both for households’ and individuals’ characteristics, while not suffering from the problem of the “zero cell value”. The results indicate that there is major day-to-day variability in working activity participation; in particular, on Wednesdays and Fridays the number of households working part-time or not working is particularly high. This is consistent with what was expected by the analysts. Working participation is a fundamental component in activity based models where work is considered a skeletal activity. The tools developed here can be useful to study how changes in population characteristics (i.e. increases of flexible working arrangements and of the number of women in the work force) affect activity participation and travel patterns.     

Microstructure and morphology of amine-cured epoxy coatings before and after outdoor exposures—An AFM study

Atomic force microscopy (AFM) has been used to study the morphology and microstructure of an amine-cured epoxy before and after outdoor exposure. Measurements were made from samples prepared in an essentially CO₂-free, H₂O-free glove box and from samples prepared in ambient conditions. For those prepared in a CO₂-free glove box, AFM imaging was conducted on (1) an unexposed air/coating surface, (2) an unexposed coating bulk, (3) an unexposed coating/substrate interface, and (4) a field exposed air/coating surface. For samples prepared in ambient conditions, only the unexposed air/coating surface was investigated. The same regions of the exposed samples were scanned periodically by the AFM to monitor changes in the surface morphology of the coating as UV exposure progressed. Small angle neutron scattering and Fourier transform infrared spectroscopy (FTIR) studies were performed to verify the microstructure and to follow chemical changes during outdoor exposure, respectively. The results have shown that amine blushing, which occurs only under ambient conditions, had a significant effect on the surface morphology and microstructure of the epoxy. The surface morphology of the samples prepared under CO₂-free, dry conditions was generally smooth and homogeneous. However, the interface and the bulk samples clearly revealed a two-phase structure consisting of bright nodular domains and dark interstitial regions, indicating an inhomogeneous microstructure. Such heterogeneous structure of the bulk was in good agreement with results obtained by small angle neutron scattering of unexposed samples and by AFM phase imaging of the degraded sample surface. The relationship between submicrometer physical changes and molecular chemical degradation is discussed. Presented at the 82nd Annual Meeting of the Federation of Societies for Coatings Technology, October 27–29, 2004, in Chicago, IL.     

Use of laser scanning confocal microscopy for characterizing changes in film thickness and local surface morphology of UV-exposed polymer coatings

Laser scanning confocal microscopy (LSCM) has been used to characterize the changes in film thickness and local surface morphology of polymer coatings during the UV degradation process. With the noninvasive feature of LSCM, one can obtain thickness information directly and nondestructively at various exposure times without destroying the specimens or deriving the thickness values from IR measurement by assuming uniform film ablation. Two acrylic polymer coatings were chosen for the study, and the physical and chemical changes of the two systems at various exposure times were measured and analyzed. Those measurable physical changes caused by UV exposure include film ablation, formation of pits and other surface defects, and increases in surface roughness. It was found in both coatings that changes in measured film thickness by LSCM were not correlated linearly to the predicted thickness loss using the changes in the CH band obtained by the Fourier Transform Infrared (FTIR) spectroscopy measurements in the later degradation stages. This result suggested it was not a uniform film ablation process during the UV degradation. At later stages, where surface deformation became severe, surface roughness and profile information using LSCM were also proven to be useful for analyzing the surface degradation process Presented at the 81st Annual Meeting of the Federation of Societies for Coatings Technology, November 13–14, 2004 in Philadelphia, PA.     

Relationship between chemical degradation and thickness loss of an amine-cured epoxy coating exposed to different UV environments

The relationship between chemical degradation and thickness loss of an unpigmented, non UV-stabilized, crosslinked amine-cured epoxy coating exposed to three UV conditions was investigated. Spin-coated samples having a thickness of approximately 7 μm on an Si substrate were prepared from a stochiometric mixture of a bisphenol A epoxy resin and a tetra-functional amine curing agent. Samples were exposed outdoors and to two accelerated laboratory UV environments. Chemical degradation and thickness loss were measured by transmission Fourier transform infrared spectroscopy (FTIRS) and laser scanning confocal microscopy (LSCM), respectively. In addition, surface roughness and morphological changes were measured by atomic forcemicrosocopy (AFM) and LSCM. Substantial chemical degradation, thickness loss, and morpholocal changes occurred in the exposed films, and the rate of chemical degradation was greater than that due to the thickness loss. This additional chemical loss was attributed to an inhomogeneous degradation process in which nanoscale localized depressions initiate at certain sites on the surface, which then enlarge and deepen with exposure time. The results of this study provide a better understanding of the degradation mechanism and should lead to the development of scientific-based models for predicting the service life of crosslinked amine-cured epoxy coatings. Presented at the 82nd Annual Meeting of the Federation of Societies for Coatings Technology, October 27–29, 2004, in Chicago, IL     

Inverse solution for a biochemical heat source in a porous medium in the presence of natural convection

The inverse problem approach by conjugate gradient with adjoint equations is adapted to the context of a non-isothermal bioprocess, controlled by internal heat generation from microbial oxidation, as could be found in a composting reactor for instance, to determine the heat source from internal temperature measurements. The volumetric heat source is assumed proportional to the rate of consumption of a substrate by a biomass, as described by a Monod model. Computations are performed for Rayleigh numbers equal to 0.25 and 25, for a representative biochemical reaction under typical boundary conditions, for constant and temperature-dependent model parameters. The influence of noisy input data is also considered. It is found that good solutions can be obtained when heat release and diffusion occur over very different time scales. The variation of the model parameters with temperature must be taken into account, but single sensor solutions are possible at relatively small Rayleigh numbers when convection is present.