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.     

IMPACCT: Methodology and Tools for Power-Aware Embedded Systems

Power-aware systems are those that must exploit a widerange of power/performance trade-offs in order to adapt to the power availabilityand application requirements. They require the integration of many novel powermanagement techniques, ranging from voltage scaling to subsystem shutdown.However, those techniques do not always compose synergistically with eachother; in fact, they can combine subtractively and often yield counterintuitive,and sometimes incorrect, results in the context of a complete system. Thiscan become a serious problem as more of these power aware systems are beingdeployed in mission critical applications.To address the problem of technique integration for power-aware embedded systems, we propose a new design tool framework called IMPACCT and the associated design methodology. The system modeling methodology includes application model for capturing timing/powerconstraints and mode dependencies at the system level. The tool performs power-awarescheduling and mode selection to ensure that all timing/power constraintsare satisfied and that all overhead is taken into account. IMPACCT then synthesizesthe implementation targeting a symmetric multiprocessor platform. Experimentalresults show that the increased dynamic range of power/performance settingsenabled a Mars rover to achieve significant acceleration while using lessenergy. More importantly, our tool correctly combines the state-of-the-arttechniques at the system level, thereby saving even experienced designersfrom many pitfalls of system-level power management.     

The renal and hepatic distribution of Bence Jones proteins depends on glycosylation: a scintigraphic study in rats

The aim of the present study was to evaluate renal and liver distribution of two monoclonal immunoglobulin light chains. The chains were purified individually from the urine of patients with multiple myeloma and characterized as lambda light chains with a molecular mass of 28 kDa. They were named BJg (high amount of galactose residues exposed) and BJs (sialic acid residues exposed) on the basis of carbohydrate content. A scintigraphic study was performed on male Wistar rats weighing 250 g for 60 min after i.v. administration of 1 mg of each protein (7.4 MBq), as the intact proteins and also after carbohydrate oxidation. Images were obtained with a Siemens gamma camera with a high-resolution collimator and processed with a MicroDelta system. Hepatic and renal distribution were established and are reported as percent of injected dose. Liver uptake of BJg was significantly higher than liver uptake of BJs (94.3 vs 81.4%) (P < 0.05). This contributed to its greater removal from the intravascular compartment, and consequently lower kidney accumulation of BJg in comparison to BJs (5.7 vs 18.6%) (P < 0.05). After carbohydrate oxidation, there was a decrease in hepatic accumulation of both proteins and consequently a higher renal overload. The tissue distribution of periodate-treated BJg was similar to that of native BJs: 82.7 vs 81.4% in the liver and 17.3 vs 18.6% in the kidneys. These observations indicate the important role of sugar residues of Bence Jones proteins for their recognition by specific membrane receptors, which leads to differential tissue accumulation and possible toxicity.     

Real-time processing and compression of DNA microarray images.

In this paper, we present a pipeline architecture specifically designed to process and compress DNA microarray images. Many of the pixilated image generation methods produce one row of the image at a time. This property is fully exploited by the proposed pipeline that takes in one row of the produced image at each clock pulse and performs the necessary image processing steps on it. This will remove the present need for sluggish software routines that are considered a major bottleneck in the microarray technology. Moreover, two different structures are proposed for compressing DNA microarray images. The proposed architecture is proved to be highly modular, scalable, and suited for a standard cell VLSI implementation.     

Influence of thermal boundary conditions on natural convection in a square enclosure partially heated from below

Natural convection in air-filled 2D square enclosure heated with a constant source from below and cooled from above is studied numerically for a variety of thermal boundary conditions at the top and sidewalls. Simulations are performed for two kinds of lengths of the heated source, i.e., a small and a large source corresponding to 20% and 80% of the total length of the bottom wall, respectively. The Rayleigh number varied from 10³ to 10⁷. Results are presented in the form of streamline and isotherm plots as well as the variation of the Nusselt number and maximum temperature at the heat source surface. Comparisons among the different thermal configurations considered are reported.     

Integration of Data from Liquid–Liquid Phase Separation Databases Highlights Concentration and Dosage Sensitivity of LLPS Drivers

Liquid–liquid phase separation (LLPS) is a molecular process that leads to the formation of membraneless organelles, representing functionally specialized liquid-like cellular condensates formed by proteins and nucleic acids. Integrating the data on LLPS-associated proteins from dedicated databases revealed only modest agreement between them and yielded a high-confidence dataset of 89 human LLPS drivers. Analysis of the supporting evidence for our dataset uncovered a systematic and potentially concerning difference between protein concentrations used in a good fraction of the in vitro LLPS experiments, a key parameter that governs the phase behavior, and the proteomics-derived cellular abundance levels of the corresponding proteins. Closer scrutiny of the underlying experimental data enabled us to offer a sound rationale for this systematic difference, which draws on our current understanding of the cellular organization of the proteome and the LLPS process. In support of this rationale, we find that genes coding for our human LLPS drivers tend to be dosage-sensitive, suggesting that their cellular availability is tightly regulated to preserve their functional role in direct or indirect relation to condensate formation. Our analysis offers guideposts for increasing agreement between in vitro and in vivo studies, probing the roles of proteins in LLPS.     

Inclined hot-wire response equations for a flow field having a dominant tangential velocity component

Modified sets of response equations have been derived for a single inclined hot-wire probe introduced in various orientations in a flow field having a dominant tangential velocity component. By means of these equations, the components of the three dimensional mean velocity vectors, the turbulence intensity and the Reynolds stresses can be easily determined at any location in a swirling flow which is stationary with time. This method is particularly applicable to the characterization of the flow field in chemical processing equipment where this type of swirling flow is increasingly being used. It has been successfully applied to a study of the flow field in a spray-drying chamber, in the course of which good reproducibility and consistency of results were obtained.     

Estimation of diffusion coefficient for solute–polymer systems

Diffusion coefficients are needed for the analysis of many mass transfer problems involving polymers. Since the diffusivity for such systems are strong functions of temperature and concentration, the analysis of these problems is greatly facilitated if predictive methods are available for the determination of the required diffusion coefficient. Because of the limitations of the theoretical approaches for estimating diffusivity, empirical correlation of solute–polymer diffusion coefficient data with physical properties of the solute were investigated. Chemical permeation measurements were made for several organic liquids through three elastomers (polychoroprene, butyl and nitrile rubber) at five different temperatures, ranging from 25 to 65°C. The collected diffusivities were correlated with liquid kinematic viscosity. Diffusivities (at different temperatures) depend mainly on the kinematic viscosity of solutes. The results also indicate a close relation between the variation of diffusivity and viscosity with temperature.     

Design and evaluation of a high throughput QoS-aware and congestion-aware router architecture for Network-on-Chip

This paper proposes a novel QoS-aware and congestion-aware Network-on-Chip architecture that not only enables quality-oriented network transmission and maintains a feasible implementation cost but also well balance traffic load inside the network to enhance overall throughput. By differentiating application traffic into different service classes, bandwidth allocation is managed accordingly to fulfill QoS requirements. Incorporating with congestion control scheme which consists of dynamic arbitration and adaptive routing path selection, high priority traffic is directed to less congested areas and is given preference to available resources. Simulation results show that average latency of high priority and overall traffic is improved dramatically for various traffic patterns. Cost evaluation results also show that the proposed router architecture requires negligible cost overhead but provides better performance for both advanced mesh NoC platforms.