Combinatorial synthesis of new cationic lipids and high-throughput screening of their transfection properties

Here we describe the first synthesis-screening approach for the identification and optimization of new cationic lipids for gene transfer in various cell lines. Combinatorial solid-phase chemistry was used to synthesize a library of new cationic lipids based on 3-methylamino-1,2-dihydroxypropane as the polar, cationic lipid part. As the nonpolar lipid part, different hydrocarbon chains were bound to the amino group of the scaffold and the amino group was further methylated to afford constantly cationic lipids. Lipids were synthesized in both configurations and as racemates, and the counter ions were also varied. By using a fully automated transfection screening method and COS-7 cells, the cationic lipid N,N-ditetradecyl-N-methyl-amino-2,3-propanediol (KL-1-14) was identified as a candidate lipid for the development of an improved transfection reagent. Screening the transfection properties of KL-1-14 in numerous combinations with the helper lipids dioleoylphosphatidylethanolamine (DOPE) and cholesterol (Chol) revealed that Chol is the most suitable helper lipid and the best KL-1-14/Chol ratio is 0.5-0.7. Compared to the standard transfection lipid N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl sulfate (DOTAP), transfection efficiency was improved by a factor of about 40. Furthermore, by using R- and S-configured KL-1-14, it could be shown that the configuration of the lipids had no significant influence on its transfection efficiency. The highest transfection efficiencies were achieved with chloride as the counter ion. The new lipofection reagent was further tested to transfect the cell lines MDA-MB-468, MCF-7, MDCK-C7, and primary dentritic cells (DC), which are important for the development of new anticancer gene therapy strategies. Even in these cells, KL-1-14/Chol (1:0.6) had improved transfection efficiencies, which were about two to four times higher than for DOTAP.     

A Unified Model for the Mixing of Non‐Newtonian Fluids in the Laminar,Transition, and Turbulent Region

Summary: Non‐Newtonian fluid behavior has significant influence on quantities in chemical engineering like power input, mixing time, heat transfer etc. In the laminar flow region, the concept of effective viscosity by Metzner and Otto is well established. In the transition region between laminar and turbulent flow, the existing concepts use three and even more empirical parameters to determine the specific power input. Here, a unified and general but simple approach is introduced to calculate the power input for shear thinning fluids over the whole flow region using just one empirical parameter. The Metzner‐Otto relation is obtained as a limiting case for the laminar region. The empirical parameter of the new approach is related to the Metzner‐Otto constant. The concept is validated for eight different stirrer systems. Mixing time and maximum shear rate and heat transfer can also be calculated using this approach. The new concept presented should also be applicable for other apparatuses, e.g., static mixers.

Comparison of experimental data and a curve calculated according to the new method (solid line).     

Surface diffusion in microporous media

The surface diffusion of He, Ne, H₂, Ar, O₂, N₂ and CO₂ through Vycor microporous glass was analyzed theoretically and experimentally in the low pressure range as a function of temperature. A statistical-mechanical treatment was carried out in terms of partition functions to derive a general equation of surface flow. It was possible to separate the surface diffusion from the total flow using a simplified working equation. The conventional method which assumes that there is no adsorbed flow for helium should be discarded, because the fraction of surface diffusion for helium ranges from 0.133 to 0.247. It is also illustrated how to predict the permeability of a new gas for the same porous medium.     

Skin detection in video under uncontrolled illumination

Multimedia Tools and Applications - Many vision-based human-computer interaction (HCI) applications require skin detection. However, their performance relies on accuracy in detecting skin regions...     

Effect of processing techniques and residual solvent on the thermal/mechanical properties of epoxy-cellulose nanocrystal nanocomposites

Epoxy nanocomposites reinforced with 0–5 wt% cellulose nanocrystal (CNC) were fabricated by solution casting method. The CNCs were first dispersed in the hardener together with a co-solvent and subsequently mixed with epoxy monomer using two different protocols: bulk mixing and dropwise mixing. The dropwise mixing resulted in more homogeneously-dispersed CNCs and provided better mechanical properties, particularly higher tensile strength, tensile modulus, and work of fracture with increasing CNC content. Investigation of various CNC/hardener mixtures with different solvent content showed that the residual solvent in the composite mixture had an impact on the curing behavior of the nanocomposite epoxy and may have lowered its crosslinking density, leading to T_g depression. However, the presence of the solvent was shown to be essential for the preparation of a well-dispersed CNC phase in the hardener solution, and to form homogenous composite mixtures. Our results show that the reinforcement effect due to higher CNC concentration is more significant than the solvent effect in the hardener system and shows greater improvement in mechanical properties. The CNC component reversed the solvent plasticizing effect through its superior mechanical reinforcing effects, and the dropwise mixing process led to better dispersion compared with the bulk mixing process.     

Influence of motion correction on the visual analysis of cardiac magnetic resonance stress perfusion imaging

Objective

Image post-processing corrects for cardiac and respiratory motion (MoCo) during cardiovascular magnetic resonance (CMR) stress perfusion. The study analyzed its influence on visual image evaluation.

Materials and methods

Sixty-two patients with (suspected) coronary artery disease underwent a standard CMR stress perfusion exam during free-breathing. Image post-processing was performed without (non-MoCo) and with MoCo (image intensity normalization; motion extraction with iterative non-rigid registration; motion warping with the combined displacement field). Images were evaluated regarding the perfusion pattern (perfusion deficit, dark rim artifact, uncertain signal loss, and normal perfusion), the general image quality (non-diagnostic, imperfect, good, and excellent), and the reader’s subjective confidence to assess the images (not confident, confident, very confident).

Results

Fifty-three (non-MoCo) and 52 (MoCo) myocardial segments were rated as ‘perfusion deficit’, 113 vs. 109 as ‘dark rim artifacts’, 9 vs. 7 as ‘uncertain signal loss’, and 817 vs. 824 as ‘normal’. Agreement between non-MoCo and MoCo was high with no diagnostic difference per-patient. The image quality of MoCo was rated more often as ‘good’ or ‘excellent’ (92 vs. 63%), and the diagnostic confidence more often as “very confident” (71 vs. 45%) compared to non-MoCo.

Conclusions

The comparison of perfusion images acquired during free-breathing and post-processed with and without motion correction demonstrated that both methods led to a consistent evaluation of the perfusion pattern, while the image quality and the reader’s subjective confidence to assess the images were rated more favorably for MoCo.

     

Flexible Furnace Concepts for Vacuum Heat Treatment Combined with High-pressure Gas Quenching

IN the past five years the process combination of vacuum hardening, respectively vacuum carburizing with high-pressure gas quenching was successfully introduced to the market, especially in the manufacture of gears. In the meantime furnace concepts for various applications are available to the industry. In the following report three plant varieties are introduced, which differ in process flexibility and throughput. This report also explains criteria for the selection of a furnace in view of the existing application requirements. Besides this a short introduction is given into the vacuum carburizing process and the high-pressure gas quenching technology.     

Flow profile measurement in microchannel using the optical feedback interferometry sensing technique

The need to accurately measure flow profiles in microfluidic channels is well recognised. In this work, we present a new optical feedback interferometry (OFI) flow sensor that accurately measures local velocity in fluids and enables reconstruction of a velocity profile inside a microchannel. OFI is a self-aligned interferometric technique that uses the laser as both the transmitter and the receiver thus offering high sensitivity, fast response, and a simple and compact optical design. The system described here is based on a commercial semiconductor laser and has been designed to achieve a micrometer-range spatial resolution. The sensor performance was validated by reconstructing the velocity profile inside a circular cross-section flow-channel with 320  $\upmu $ m internal diameter, with a relative error smaller than 1.8 %. The local flow velocity is directly measured, thus avoiding the need for model based profile calculation and uncertainties inherent to this approach. The system was validated by successfully extracting the flow profiles in both Newtonian and shear-thinning liquids.