Recent Biophysical Issues About the Preparation of Solute-Filled Lipid Vesicles

Here we report some recent biophysical issues on the preparation of solute-filled lipid vesicles and their relevance to the construction of “synthetic cells.” First, we introduce the “semi-synthetic minimal cells” as the liposome-based cell-like systems, which contain a minimal number of biomolecules required to display simple and complex biological functions. Next, we focus on recent aspects related to the construction of synthetic cells. Emphasis is given to the interplay between the methods of synthetic cell preparation and the physics of solute encapsulation. We briefly introduce the notion of structural and compositional “diversity” in synthetic cell populations.     

Preparation and characterization of chitosan‐based dispersions

Complexation of chitosan in aqueous solutions by low molecular weight electrolytes is one of the simplest methods for the preparation of aqueous chitosan dispersions. In this work, the influence of storage time, sulfate concentration, method of preparation and surfactant content on some properties of the resultant chitosan dispersions (turbidity, viscosity and zeta potential) was analyzed. Turbidimetry was adequate to monitor the formation of particles, while viscometry was suitable to monitor changes in the dispersing phase. An analysis of the properties of these systems, mainly in terms of particle–particle and macromolecule–macromolecule interactions was carried out. Copyright © 2004 Society of Chemical Industry     

A two-layer architecture for economically optimal process control and operation

A two-layer architecture for dynamic real-time optimization (or nonlinear modelpredictive control (NMPC) with an economic objective) is presented, where the solution of the dynamic optimization problem is computed on two time-scales. On the upper layer, a rigorous optimization problem is solved with an economic objective function at a slow time-scale, which captures slow trends in process uncertainties. On the lower layer, a fast neighboring-extremal controller is tracking the trajectory in order to deal with fast disturbances acting on the process. Compared to a single-layer architecture, the two-layer architecture is able to address control systems with complex models leading to high computational load, since the rigorous optimization problem can be solved at a slower rate than the process sampling time. Furthermore, solving a new rigorous optimization problem is not necessary at each sampling time if the process has rather slow dynamics compared to the disturbance dynamics. The two-layer control strategy is illustrated with a simulated case study of an industrial polymerization process.     

Anthropomorphisms in multimedia learning: Attract attention but do not enhance learning?

Anthropomorphizing graphical elements in multimedia learning materials improves learning outcomes. The reasons for enhanced learning are unclear. We extended a seminal anthropomorphism study in order to examine whether the effect of anthropomorphisms on learning outcomes, both immediate and delayed, is caused by the anthropomorphized elements' effects on attention distribution or by elevated positive affective–motivational states. The study had a partial 3 × 2 design (the materials' graphics: schematic vs. black‐and‐white anthropomorphisms vs. colourful anthropomorphisms × eye tracker: present vs. absent). The participants were university students (N = 181). Unexpectedly, we found no significant effect of anthropomorphisms on learning outcomes. Anthropomorphisms significantly affected attention distribution during initial fixations but not overall. Modest effect on enjoyment was found, but no such effect was detected as concerns flow and generalized positive affect. We also found that the eye tracker's mere presence had slight adverse effects on learners, but these effects did not compromise learning.     

Active application of primer acid on acid‐treated enamel: Influence on the bond effectiveness of self‐etch adhesives systems

Aim: Evaluate the composite‐to‐enamel bond after passive or active application of self‐etching primer systems on polished or pre‐etched enamel with phosphoric acid. Materials and Methods: Two self‐etch adhesives systems (SEAS) were used: Clearfil SE Bond and Easy Bond. Third human molars were divided into 8 groups (N = 10). The crown of each tooth was sectioned into halves and the mesial/distal surfaces were used. The adhesives were actively or passively applied on enamel with or without prior phosphoric‐acid etching. Resin composite cylinders were built after adhesive application. After stored in relative humidity for 24 hr/37°C the specimens were subjected to microshear test in universal testing a machine at a crosshead speed of 0.5 mm/minute. The results were analyzed with three‐way ANOVA and the Tukey test. The enamel‐etching pattern was evaluated under SEM. Results: The 2‐step SEAS system presented significantly higher adhesive bond strength means (47.37 MPa) than the 1‐step (36.87 MPa). A poor enamel‐ etching pattern was observed in active mode showing irregular and short resin tags, however there was not compromised the bond strength. Conclusion: Active or passive application produced similar values of bond strength to enamel regardless of enamel pretreatment and type of SEAS.     

A robotic DNA purification protocol and real-time PCR for the detection of Campylobacter jejuni in foods

Primers designed to amplify a Campylobacter jejuni cadF gene sequence were used in an SYBR Green I real-time PCR assay as an alternative to conventional bacteriological methods for the rapid detection of C. jejuni in foods. Twenty-five grams of chicken skin (breast and thigh) was contaminated by adding approximately 1, 10, or 50 CFU of C. jejuni ATCC 35560. Twenty-five grams of pork and 25-ml aliquots of milk were also inoculated with 1 and 10 CFU of the pathogen. The samples were incubated in Bolton broth for different periods at 37 and 42 degrees C under microaerophilic conditions. Using a commercial robotic DNA purification system, DNA was extracted and purified from 1-ml aliquots of the enrichment cultures before and after centrifugation of the 250-ml enrichment broth at 15,900 x g for 10 min at 4 degrees C. The DNA was used as the template in a real-time PCR assay. C. jejuni was detected after 12 h of enrichment from samples inoculated with about 50 CFU/25-g sample. After centrifugation, an enrichment step of 8 h was sufficient to allow detection of pathogen in samples inoculated with 10 CFU/25 g. However, 24 h of enrichment was necessary to detect pathogen in samples inoculated with approximately 1 CFU/25 g. The real-time PCR protocol developed in this study significantly reduced the detection time of C. jejuni in foods.     

Survey and comparison of message authentication solutions on wireless sensor networks

Security is an important concern in any modern network. This also applies to Wireless Sensor Networks (WSNs), especially those used in applications that monitor sensitive information (e.g., health care applications). However, the highly constrained nature of sensors imposes a difficult challenge: their reduced availability of memory, processing power and energy hinders the deployment of many modern cryptographic algorithms considered secure. For this reason, the choice of the most memory-, processing- and energy-efficient security solutions is of vital importance in WSNs. To date, a number of extensive analyses comparing different encryption algorithms and key management schemes have been developed, while very little attention has been given to message authentication solutions. In this paper, aiming to close this gap, we identify cipher-based Message Authentication Codes (MACs) and Authenticated Encryption with Associated Data (AEAD) schemes suitable for WSNs and then evaluate their features and performance on a real platform (TelosB). As a result of this analysis, we identify the recommended choices depending on the characteristics of the target network and available hardware.     

Functional display of active bovine adrenodoxin on the surface of E. coli by chemical incorporation of the [2Fe-2S] cluster

The display of heterologous proteins on the surface of living cells bears promising options for a wide variety of biotechnological applications. Up to now, however, cellular surface display was merely restricted to simple polypeptide chains. Here we present for the first time the efficient display of a protein (bovine adrenodoxin) that contains an inorganic, prosthetic group in its active form on the surface of Escherichia coli. For this purpose apo-adrenodoxin was transported to the cell surface and anchored within the outer membrane by the autotransporter pathway. Incorporation of the iron-sulfur cluster was achieved by a single-vial, one-step titration under anaerobic conditions. The biological function of surface-displayed holo-adrenodoxin could be established through adrenodoxin-dependent steroid conversion by two different cytochrome P450 enzymes and the number of functional molecules on the cell surface could be determined to be more than 10(5) per cell. Neither the expression of adrenodoxin nor the incorporation of the chemical iron-sulfur cluster reduced the viability of the bacterial cells.