Hyperthermostable mutants of Bacillus licheniformis {alpha}-amylase: multiple amino acid replacements and molecular modelling

We have identified previously two critical positions for thethermostability of the highly thermostable -amylase from Bacilluslicheniformis. We have now introduced all 19 possible aminoacid residues to these two positions, His 133 and Ala209. Themost favourable substitutions were to Ile and Val, respectively,which both increased the half-life of the enzyme at 80°Cby a factor of 3. At both positions a stabilizing effect ofhydrophobic residues was observed, although only in the caseof position 133 could a clear correlation be drawn between thehydrophobicity of the inserted amino acid and the gain in proteinstability. The construction of double mutants showed a cumulativeeffect of the most favourable and/or deleterious substitutions.Computer modelling was used to generate a 3-D structure of thewild-type protein and to model substitutions at position 209,which lies in the conserved (/ß)₈ barrel domain of-amylase; Ala209 would be located at the beginning of the thirdhelix of the barrel, in the bottom of a small cavity facingthe fourth helix. The model suggests that replacement by, forexample, a valine could fill this cavity and therefore increaseintra- and interhelical compactness and hydrophobic interactions.     

High‐Throughput Differentiation of Embryonic Stem Cells into Cardiomyocytes with a Microfabricated Magnetic Pattern and Cyclic Stimulation

Pluripotent stem cells are central tools to many regenerative medicine strategies due to their ability to differentiate toward the three embryonic germ layers. One challenge remains in providing control over their differentiation into specific lineages, such as cardiac commitment. Here, the possibility of directing cardiomyogenesis of embryonic stem cells using a microfabricated magnetic pattern is demonstrated. The stem cells are labeled with magnetic nanoparticles, aggregated into embryoid bodies (EBs) onto the pattern, and stimulated with a local magnetic force applied via the pattern. The EBs formed on this magnetic device experience the same differentiation profile as the ones created by the common hanging drop approach, while it allows high‐throughput production of hundreds of EBs. Further on/off cyclic magnetic force stimulation mediated by the same device is sufficient to enhance cardiomyogenesis in a way that almost all EBs develop spontaneous beating, confirmed by the overexpression of α‐actin and troponin proteins, and by the upregulation (twofold to fivefold) of genes involved in mesoderm differentiation (Nkx2.5, Gata4, and Gata6), and more specifically cardiac lineage (Tnnt2, Myh6, and Myl‐2). Beyond holding high application‐level potential, this work confirms that physical forces, and specifically on/off dynamic ones can be sufficient to govern cell function.     

Low temperature solid–liquid interdiffusion wafer and die bonding based on PVD thin Sn/Cu films

Microsystem Technologies - Cu–Sn intermetallic diffusion has been studied with thin Sn and Cu films deposited by physical vapor deposition. It was found that annealing time is critical to the...     

Planetary LakeLander—A Robotic Sentinel to Monitor Remote Lakes

This field report describes the design and operations of the Planetary LakeLander (PLL) probe and its ground data systems. LakeLander's primary mission is to characterize the physical, chemical, and biological processes occurring in a high‐altitude lake, and how they are being impacted by rapid deglaciation. LakeLander's secondary purpose is to test operation concepts for future exploration of Titan's lakes. The LakeLander probe is a permanently anchored buoy that measures both surface meteorology and water quality parameters in the top 40 m of the water column. The concept of operations calls for the probe to continue collecting and downlinking data through the Andean winter, when the lake is inaccessible; this drives the power system design and forces a strong focus on system reliability, analogous to a space mission. The PLL ground data system provides the central archive of downlinked data. They are structured around a unified data‐sharing web site that includes tools for mapping, data visualization, documentation, and numerical analysis. The web site provides a hub for engaging the science team and enables interdisciplinary collaboration. This report concludes with lessons learned during field deployment and several months of remote operations on the lake. Among the conclusions: (1) the choice to use an off‐the‐shelf profiling system has proven wise; (2) effective maintenance of a long‐lived remote system requires extensive measurement, logging, and display of as many system variables as possible; and (3) the visualization sandbox component of the data‐sharing web site has made numerical analysis of probe data much easier and more accessible to the entire interdisciplinary science team.     

Improving kriging surrogates of high-dimensional design models by Partial Least Squares dimension reduction

Engineering computer codes are often computationally expensive. To lighten this load, we exploit new covariance kernels to replace computationally expensive codes with surrogate models. For input spaces with large dimensions, using the kriging model in the standard way is computationally expensive because a large covariance matrix must be inverted several times to estimate the parameters of the model. We address this issue herein by constructing a covariance kernel that depends on only a few parameters. The new kernel is constructed based on information obtained from the Partial Least Squares method. Promising results are obtained for numerical examples with up to 100 dimensions, and significant computational gain is obtained while maintaining sufficient accuracy.     

Study of the correlation between flexible food packaging peeling resistance and surface composition for aluminum-metallized BOPP films aged at 60°C

ABSTRACT

This study investigated the correlation between surface composition and peeling resistance in food packaging films by studying the heat aging of fabricated films over varying periods of time. The films consisted of a layer of aluminum (Al) metallized, biaxially oriented polypropylene (BOPP) bonded with a polyurethane (PU) adhesive onto another polymeric layer of low-density polyethylene (LDPE). The Al metallized films were prepared by physical vapor deposition (PVD) and aged at 60°C for either 5 or 15 days. The resulting aluminum surfaces were analyzed using X-ray photoelectron spectroscopy (XPS) and found to contain aluminum oxide (Al₂O₃) and trihydroxide (Al(OH)₃). The XPS characterization also revealed a 29% increase in the Al(OH)₃ layer thickness of the aged sample relative to a non-aged sample. Atomic force microscopy (AFM) was applied on investigations of possible morphology changes. The aluminum and PU adhesive surface energies were also determined using contact angles measurements and the aluminum surface energy was found to increase by as much as 11.7% compared to the non-aged sample, while the PU adhesive surface energy was at least 65% higher than that of the metallic substrate. The peeling resistance of the laminated aluminum was determined by average peel strength measurements and it was found that the variation in peel strength was related to changes in the Al₂O₃ layer thickness. The delaminated samples were analyzed using scanning electron microscopy/energy dispersive X-ray spectrometry (SEM/EDS) and showed the cohesive failure of the aluminum film.     

pH effects on catalytic ozonation of carboxylic acids with metal on metal oxides catalysts

Summary The pH influence on the catalytic ozonation of small carboxylic acids (succinic (SA), chloroacetic (CAA) and pyruvic (PA) acids) was studied and compared to ozonation alone. In the acidic systems, the conversion of these carboxylic acids and total organic carbon (TOC) during the catalytic ozonation is mainly due to the catalytic effect. Experiments on SA indicate that in the pH range 3.6–5.0, in contrast to the ozone behavior, the efficiency of the catalytic ozonation system is decreased with increasing pH due to the change of charges on the catalyst surface. In the basic pH range (7.2–10.0), the effect of the ozonation alone becomes important and the reaction in the presence of the catalyst is a combined effect of ozonation alone and catalytic oxidation. Although the apparent SA conversion by the catalytic ozonation in the basic systems is higher than in the acidic systems, the Total Organic Carbon conversion is not so much higher than in the latter. During the ozonation with the same catalyst, more than 80 of the CAA converted are mineralized. The oxidation of CAA leads to almost equimolar release of chloride ions. This ion has no detrimental influence on the catalytic effects under our experimental conditions. However, in the very dilute systems with pH value lower than the pKa of CAA, when the CAA concentration is decreased to about 0.6mM, the catalytic oxidation is almost stopped. The low concentration of the dissociated CAA induces only weak adsorption and consequently small reaction rates. The total elimination of CAA with low concentration can be reached only if the pH is increased above the pKa. In addition to the distinct pH effect in the ozonation and catalytic ozonation system observed in this work, a further demonstration that separate mechanisms are involved in the two processes is provided by the analysis of oxidation by-products of pyruvic acid.     

Use Of Static Mixer For Oxidation And Disinfection By Ozone

Ozone is used in drinking water treatment as a biocide, as an oxidant and as a pretreatment in order to improve the performance of subsequent processes. Increasing concern over the quality of drinking water has led to a number of new stringent regulations in the control of chemical and microbiological contaminants. Disinfection deals with the concept of “CT”, which is the need to maintain a certain minimum concentration for a given time. Under ideal laboratory conditions, it is 0.4 mg O₃/L for 4 min. In practice, since the method for the CT determination has not been finalized by the EPA, “T” can be the minimum detention time of 90% of total flow, and “C” can be a measured ozone residual at the outlet of cells of the contactor. New standards for micropollutants in drinking water imply an optimization of the ozonation step, by improving the ozone transfer from gas to water, and the control of the detention time as well as ozone residual within the contactor.

All these considerations have led us to use static mixers to transfer ozone into water. This process enables us to control the ozone concentration in water and detention time. It is a very simple system, with very low maintenance requirements due to the lack of moving parts. Civil engineering is minimized. A pilot scale study is presented here. It took place at the Méry-sur-Oise water treatment plant, on a pilot plant working at 8-12 m³/h. It is composed of a static mixer for the transfer of ozone from gas to liquid, linked to an air lift to separate gas from liquid, providing ozonated water.

The optimization of transfer was achieved by studying the impact of water flow, gas flow and ozone concentration in the gas. It is possible to reach 90% of transfer in less than 15 s. Headloss (ΔP) across the mixer is a function of gas and water flows and remains economically very acceptable as 0.15 bar for 12 m³/h.

Atrazine removal was studied using a static mixer, an air lift and a contact pipe 80-m long, providing an optimum contact time phase, working as a plug flow reactor. Ozone and H₂O₂/O₃ treatments were compared. The maximum reduction of atrazine concentrations (e.g., for an infinite contact time) is a function of the amount of transferred ozone, but H₂O₂ influences the kinetics of the reaction. In the presence of H₂O₂ with a ratio of H₂O₂ to O₃ of 0.4 w/w, maximum elimination is reached in 2 min 30 s.

The effect of such treatments on environmental bacteria also was followed. A counting of total germs at 20°C showed a decrease of 1- to 3-logs 10 after 1 min 30 s of contact time for about 2 mg/L of transferred ozone. No significant difference between treatments with or without H₂O₂ was shown. The same conclusions were obtained from heterotrophic plate counts (37°C) and epifluorescence countings.     

Properties of a thermotropic liquid crystalline polymer blended with different thermoplastics

Thermal, rheological, morphological, and mechanical properties of a thermotropic liquid crystalline polymer, TLCP (copolyester Vectra A-950 from Hoechst), blended with a polycarbonate (PC), a polyethylene glycol terephthalate (PETG), and a blend of PC and PETG (20/80) are presented and discussed. Important supercooling effects are observed for the TLCP. For the blends the glass transition temperature of the matrix is shown to decrease slightly, suggesting partial miscibility of the components. A finer dispersion is observed for the TLCP/PC blends, at least for TLCP concentrations lower than 20%, for which the mechanical properties are quite good. For higher TLCP concentrations, as well as for the other two matrices, the mechanical properties follow more or less the mixing rule, and the morphology of the blends suggests poor adhesion. We were unable to obtain fibrillar structures by extruding the blends through a capillary rheometer; in the TLCP/PC blends, the TLCP domains were too small, and for the other blends the extrudates had not enough melt strength.