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.     

Linear friction welding of Ti6Al4V: experiments and modelling

Linear friction welding of the Ti6Al4V alloy is studied. A new definition of the energy input rate is proposed, based on an integration over time of the in-plane force and velocity; a strong correlation with the upset rate is then found. The effective friction coefficient is estimated to be 0·5±0·1 for varying frequencies and amplitudes, with only a weak dependence on the processing conditions displayed. A model is proposed that accounts for both the conditioning and equilibrium stages of the process, which is shown to be in good agreement with the experimental data. The model is used to study the mechanism by which the flash is formed. A criterion is proposed by which the rippled nature of its morphology can be predicted.     

EEG spectral abnormalities and psychosis as predictors of cognitive and functional decline in probable Alzheimer's disease

We examined whether either psychotic features (e.g., delusions and hallucinations) or EEG abnormalities are associated with more rapid progression of Alzheimer's disease (AD). AD patients with psychosis have exhibited more EEG abnormalities than those without psychosis, and both abnormal EEG and psychosis have been noted to be predictors of functional and cognitive decline in AD. Ninety-five probable AD patients participating in a longitudinal study of dementia had an EEG and a semistructured psychiatric interview at baseline. Using EEG spectral analysis, we classified records as normal/abnormal based on the parasagittal mean frequency. Patients with abnormal EEGs were more functionally (e.g., Blessed Rating Scale for activities of daily living) and cognitively (e.g., Mini-Mental State) impaired than patients with normal EEG. AD patients with psychosis were only more functionally impaired than patients without psychosis. A two-factor analysis showed no interaction between abnormal EEG and psychosis. In addition, using a Cox proportional hazard model adjusted for age and education, the presence of an abnormal EEG or psychotic symptom at study entry was associated with higher risk of reaching severe cognitive and functional impairment during follow-up. Neither abnormal EEG nor the presence of psychosis predicted death. These results indicate that both abnormal EEG and psychosis are independent predictors of disease progression but not of physical survival.     

Mixed Mode VLSI Implementation of a Neural Associative Memory

A mixed mode digital/analog special purpose VLSI hardware implementation of an associative memory with neural architecture is presented. The memory concept is based on a matrix architecture with binary storage elements holding the connection weights. To enhance the processing speed analog circuit techniques are applied to implement the algorithm for the association. To keep the memory density as high as possible two design strategies are considered. First, the number of transistors per storage element is kept to a minimum. In this paper a circuit technique that uses a single 6-transistor cell for weight storage and analog signal processing is proposed. Second, the device precision has been chosen to a moderate level to save area as much as possible. Since device mismatch limits the performance of analog circuits, the impact of device precision on the circuit performance is explicitly discussed. It is shown that the device precision limits the number of rows activated in parallel. Since the input vector as well as the output vector are considered to be sparsely coded it is concluded, that even for large matrices the proposed circuit technique is appropriate and ultra large scale integration with a large number of connection weights is feasible.     

On the Energy Landscape at the Glass Transition

A recent hypothesis claims that the glass transition itself, though it is a very pronounced relaxation peak, is no separate relaxation process at all, but is just the breakdown of the shear modulus due to the weak elastic dipole interaction between all the quasi-independent relaxation centers of the glass. Two derivations are considered, one of them in terms of a breakdown of the shear modulus and the second in terms of a divergence of the shear compliance. Mechanical relaxation data from the literature for vitreous silica, glycerol, polymethylmethacrylate and polystyrene are found to be consistent with the first hypothesis. PACS numbers: 64.70.Pf     

Interfacial hybridization kinetics of oligonucleotides immobilized onto fused silica surfaces

Fused silica optical fibers have been used in an intrinsic mode optical configuration as biosensors for fluorescence based detection of hybridization of nucleic acids. In this work, the kinetics of hybridization of single-stranded oligonucleotides that were covalently immobilized were studied. The probe DNA was dT₂₀, and the target was Fluorescein-labeled non-complementary (dT₂₀) or complementary (dA₂₀) oligonucleotide. Chronofluorimetric monitoring of the adsorption and hybridization processes was used to investigate oligonucleotide films of different density, in different salt concentrations, at temperatures of 25 and 40 °C, with the concentration of the target DNA being 0.005–0.1 μM. Mathematical models based on first- and second-order Langmuir adsorption have been examined to describe both the adsorption and the hybridization processes. Experimental data were processed using the models, and the hybridization kinetics were calculated. Hybridization kinetics on these optical fiber DNA sensors was found to be up to three orders faster than results presented for a number of other experiments using different immobilization chemistries.     

A fictitious domain decomposition method for the solution of partially axisymmetric acoustic scattering problems. Part I: Dirichlet boundary conditions

We present a fictitious domain decomposition method for the fast solution of high‐frequency acoustic scattering problems characterized by a partially axisymmetric sound‐soft scatterer. We apply this method to the solution of various mockup submarine problems, and highlight its computational advantages and intrinsic parallelism. Copyright © 2002 John Wiley & Sons, Ltd.