Improved photobiological H-2 production in engineered green algal cells

Oxygenic photosynthetic organisms use solar energy to split water (H2O) into protons (H+), electrons (e(-)), and oxygen. A select group of photosynthetic microorganisms, including the green alga Chlamydomonas reinhardtii, has evolved the additional ability to redirect the derived H+ and e(-) to drive hydrogen (H-2) production via the chloroplast hydrogenases HydA1 and A2 (H(2)ase). This process occurs under anaerobic conditions and provides a biological basis for solar-driven H-2 production. However, its relatively poor yield is a major limitation for the economic viability of this process. To improve H-2 production in Chlamydomonas, we have developed a new approach to increase H+ and e(-) supply to the hydrogenases. In a first step, mutants blocked in the state 1 transition were selected. These mutants are inhibited in cyclic e(-) transfer around photosystem I, eliminating possible competition for e(-) with H(2)ase. Selected strains were further screened for increased H-2 production rates, leading to the isolation of Stm6. This strain has a modified respiratory metabolism, providing it with two additional important properties as follows: large starch reserves ( i.e. enhanced substrate availability), and a low dissolved O-2 concentration (40% of the wild type (WT)), resulting in reduced inhibition of H2ase activation. The H-2 production rates of Stm6 were 5 - 13 times that of the control WT strain over a range of conditions ( light intensity, culture time, +/- uncoupler). Typically, similar to 540 ml of H-2 liter(-1) culture ( up to 98% pure) were produced over a 10-14-day period at a maximal rate of 4 ml h(-1) ( efficiency = similar to 5 times the WT). Stm6 therefore represents an important step toward the development of future solar-powered H-2 production systems.

Using different structure types of microemulsions for the preparation of poly(alkylcyanoacrylate) nanoparticles by interfacial polymerization

A phase diagram of the pseudoternary system ethyloleate, polyoxyethylene 20 sorbitan mono-oleate/sorbitan monolaurate and water with butanol as a cosurfactant was prepared. Areas containing optically isotropic, low viscosity one-phase systems were identified and systems therein designated as w/o droplet-, bicontinuous- or solution-type microemulsions using conductivity, viscosity, cryo-field emission scanning electron microscopy and self-diffusion NMR. Nanoparticles were prepared by interfacial polymerization of selected w/o droplet, bicontinuous- or solution-type microemulsions with ethyl-2-cyanoacrylate. Morphology of the particles and entrapment of the water-soluble model protein ovalbumin were investigated. Addition of monomer to the different types of microemulsions (w/o droplet, bicontinuous, solution) led to the formation of nanoparticles, which were similar in size (similar to 250 nm), polydispersity index (similar to 0.13), zeta-potential (similar to-17 mV) and morphology. The entrapment of the protein within these particles was up to 95%, depending on the amount of monomer used for polymerization and the type of microemulsion used as a polymerization template. The formation of particles with similar characteristics from templates having different microstructure is surprising, particularly considering that polymerization is expected to occur at the water-oil interface by base-catalysed polymerization. Dynamics within the template (stirring, viscosity) or indeed interfacial phenomena relating to the solid-liquid interface appear to be more important for the determination of nanoparticle morphology and characteristics than the microstructure of the template system. (c) 2005 Elsevier B.V. All rights reserved.

Preparation of poly (alkylcyanoacrylate) nanoparticles by polymerization of water-free microemulsions

Phase diagrams of the pseudoternary systems ethyloleate, polyoxyethylene 20 sorbitan mono-oleate/sorbitan monolaurate and propylene glycol with and without butanol as a co-surfactant were prepared. Areas containing optically isotropic, one-phase systems were identified and samples therein designated as droplet, bicontinuous or solution type microemulsions using conductivity, viscosity and self-diffusion NMR. Nanoparticles were prepared by polymerization of selected microemulsions with ethyl-2-cyanoacrylate and the morphology of the particles was investigated. Addition of monomer to all types of microemulsions led to the formation of nanoparticles, which had an average size of 244 +/- 25 nm, an average polydispersity index of 0.15 +/- 0.04 and a zeta-potential of -17 +/- 3 mV. The formation of particles from water-free microemulsions of different types is surprising, particularly considering that polymerization is expected to occur at a water-oil interface by base-catalysed polymerization. It would appear that propylene glycol is sufficiently nucleophilic to initiate the polymerization. The use of water-free microemulsions as templates for the preparation of poly (alkylcyanoacrylate) nanoparticles opens up interesting opportunities for the encapsulation of bioactives which do not have suitable properties for encapsulation on the basis of water-containing microemulsions.

Affinity partitioning of plasmid DNA with a zinc finger protein

The affinity isolation of pre-purified plasmid DNA (pDNA) from model buffer solutions using native and poly(ethylene glycol) (PEG) derivatized zinc finger–GST (Glutathione-S-Transferase) fusion protein was examined in PEG–dextran (DEX) aqueous two-phase systems (ATPSs). In the absence of pDNA, partitioning of unbound PEGylated fusion protein into the PEG-rich phase was confirmed with 97.5% of the PEGylated fusion protein being detected in the PEG phase of a PEG 600–DEX 40 ATPS. This represents a 1322-fold increase in the protein partition coefficient in comparison to the non-PEGylated protein (Kc = 0.013). In the presence of pDNA containing a specific oligonucleotide recognition sequence, the zinc finger moiety of the PEGylated fusion protein bound to the plasmid and steered the complex to the PEG-rich phase. An increase in the proportion of pDNA that partitioned to the PEG-rich phase was observed as the concentration of PEGylated fusion protein was increased. Partitioning of the bound complex occurred to such an extent that no DNA was detected by the picogreen assay in the dextran phase. It was also possible to partition pDNA using a non-PEGylated (native) zinc finger–GST fusion protein in a PEG 1000–DEX 500 ATPS. In this case the native ligand accumulated mainly in the PEG phase. These results indicate good prospects for the design of new plasmid DNA purification methods using fusion proteins as affinity ligands.

Multi-phase electric drives for mobility

Multi-phase electrical drives are potential candidates for the employment in innovative electric vehicle powertrains, in response to the request for high efficiency and reliability of this type of application. In addition to the multi-phase technology, in the last decades also, multilevel technology has been developed. These two technologies are somewhat complementary since both allow increasing the power rating of the system without increasing the current and voltage ratings of the single power switches of the inverter. In this thesis, some different topics concerning the inverter, the motor and the fault diagnosis of an electric vehicle powertrain are addressed. In particular, the attention is focused on multi-phase and multilevel technologies and their potential advantages with respect to traditional technologies. First of all, the mathematical models of two multi-phase machines, a five-phase induction machine and an asymmetrical six-phase permanent magnet synchronous machines are developed using the Vector Space Decomposition approach. Then, a new modulation technique for multi-phase multilevel T-type inverters, which solves the voltage balancing problem of the DC-link capacitors, ensuring flexible management of the capacitor voltages, is developed. The technique is based on the proper selection of the zero-sequence component of the modulating signals. Subsequently, a diagnostic technique for detecting the state of health of the rotor magnets in a six-phase permanent magnet synchronous machine is established. The technique is based on analysing the electromotive force induced in the stator windings by the rotor magnets. Furthermore, an innovative algorithm able to extend the linear modulation region for five-phase inverters, taking advantage of the multiple degrees of freedom available in multi-phase systems is presented. Finally, the mathematical model of an eighteen-phase squirrel cage induction motor is defined. This activity aims to develop a motor drive able to change the number of poles of the machine during the machine operation.

Sistemas aquosos bifásicos: fundamentos e aplicações para partição/purificação de proteínas

By the year 2005 the world biochemical market will reach an estimated $ 100 billion and separation processes are a vital link between lab discoveries and the fulfillment of this commercialization potential. The practical application of aqueous two-phase systems (ATPS) to extraction processes has been exploited for several years for the recovery of biological products. Unfortunately, this has not resulted in an extensive presence of the technique in commercial processes. In this paper a critical overview of the fundamental thermodynamic properties related to formation of aqueous two-phase systems and their application to extraction and purification of bioparticules is presented.

Energy system contributions to the special judo fitness test

Purpose: This study investigated the energy system contributions of judo athletes to the Special Judo Fitness Test (SJFT). Methods: Fourteen male judo athletes performed the SJFT, which comprised three periods of judo activity (A = 15 s, B and C = 30 s) interspersed with 10 s rest intervals. During this test, one athlete threw two others positioned 6 m from each other using the ippon-seoi-nage technique. The fractions of the aerobic, anaerobic alactic and anaerobic lactic systems were calculated based on oxygen uptake, the fast component of excess postexercise oxygen uptake, and changes in net blood lactate, respectively. The contribution of the three energy systems was compared using a repeated measures analysis of variance and Bonferroni's multiple comparisons test. Compound symmetry, or sphericity, was determined by Mauchly's test. A level of significance of 5% (P < .05) was adopted in all analyses. Results: The alactic energy system presented a higher (F = 20.9; P < .001; power observed = 1.0) contribution (86.8 +/- 23.6 kJ; 42.3 +/- 5.9%) during the test when compared with both aerobic (57.1 +/- 11.3 kJ; 28.2 +/- 2.9%) and lactic (58.9 +/- 12.1 kJ; 29.5 +/- 6.2%) energy systems (P < .001 for both comparisons). Conclusions: The higher alactic contribution seems to be a consequence of the high-intensity efforts performed during the test, and its intermittent nature. Thus, when using the SJFT, coaches are evaluating mainly their athletes' anaerobic alactic system, which can be considered to be the most predominant system contributing to the actions (techniques) performed in the match.

A Novel Organotellurium Compound (RT-01) as a New Antileishmanial Agent

Leishmaniasis is a neglected disease and endemic in developing countries. A lack of adequate and definitive chemotherapeutic agents to fight against this infection has led to the investigation of numerous compounds. The aim of this study was to investigate the effect of RT-01, an organotellurane compound presenting biological activities, in 2 experimental systems against Leishmania amazonensis. The in vitro system consisted of promastigotes and amastigotes forms of the parasite, and the in vivo system consisted of L. amazonensis infected BALB/c mice, an extremely susceptible mouse strain. The compound proved to be toxic against promastigotes and amastigotes. The study also showed that treatment with RT-01 produces an effect similar to that treatment with the reference antimonial drug, Glucantime, in L. amazonensis infected mice. The best results were obtained following RT-01 intralesional administration (720 mu g/kg/day); mice showed significant delay in the development of cutaneous lesions and decreased numbers of parasites obtained from the lesions. Significant differences in tissue pathology consisted mainly of no expressive accumulation of inflammatory cells and well-preserved structures in the skin tissue of RT-01-treated mice compared with expressive infiltration of infected cells replacing the skin tissue in lesions of untreated mice. These findings highlight the fact that the apparent potency of organotellurane compounds, together with their relatively simple structure, may represent a new avenue for the development of novel drugs to combat parasitic diseases.

Simplified Mathematical Model for an Anaerobic Sequencing Batch Biofilm Reactor Treating Lipid-Rich Wastewater Subject to Rising Organic Loading Rates

This study proposes a simplified mathematical model to describe the processes occurring in an anaerobic sequencing batch biofilm reactor (ASBBR) treating lipid-rich wastewater. The reactor, subjected to rising organic loading rates, contained biomass immobilized cubic polyurethane foam matrices, and was operated at 32 degrees C +/- 2 degrees C, using 24-h batch cycles. In the adaptation period, the reactor was fed with synthetic substrate for 46 days and was operated without agitation. Whereas agitation was raised to 500 rpm, the organic loading rate (OLR) rose from 0.3 g chemical oxygen demand (COD) . L(-1) . day(-1) to 1.2 g COD . L(-1) . day(-1). The ASBBR was fed fat-rich wastewater (dairy wastewater), in an operation period lasting for 116 days, during which four operational conditions (OCs) were tested: 1.1 +/- 0.2 g COD . L(-1) . day(-1) (OC1), 4.5 +/- 0.4 g COD . L(-1) . day(-1) (OC2), 8.0 +/- 0.8 g COD . L(-1) . day(-1) (OC3), and 12.1 +/- 2.4 g COD . L(-1) . day(-1) (OC4). The bicarbonate alkalinity (BA)/COD supplementation ratio was 1:1 at OC1, 1:2 at OC2, and 1:3 at OC3 and OC4. Total COD removal efficiencies were higher than 90%, with a constant production of bicarbonate alkalinity, in all OCs tested. After the process reached stability, temporal profiles of substrate consumption were obtained. Based on these experimental data a simplified first-order model was fit, making possible the inference of kinetic parameters. A simplified mathematical model correlating soluble COD with volatile fatty acids (VFA) was also proposed, and through it the consumption rates of intermediate products as propionic and acetic acid were inferred. Results showed that the microbial consortium worked properly and high efficiencies were obtained, even with high initial substrate concentrations, which led to the accumulation of intermediate metabolites and caused low specific consumption rates.

Mn dimers on graphene nanoribbons: An ab initio study

We show, using ab initio density functional theory calculations, that Mn dimers adsorbed on graphene nanoribbons (Mn(2)/GNRs) present a magnetic bistability, as does the isolated Mn dimer. Our total energy results indicate that Mn dimers lying along the edge sites of zigzag GNRs represent the most likely configuration. We find that similar to the isolated Mn(2) molecule, the antiferromagnetic coupling represents the ground state for Mn(2)/GNR, and the spin density configuration of the GNR does not play an important role on the net magnetic moment of Mn(2), which makes GNRs an ideal substrate for adsorption of these molecules. The ground state and the excited state configuration of the Mn dimer, viz., low-spin (LS) and high-spin (HS), are maintained in the face of changes in the spin density configuration of the substrate. Here we find that the Mn(2)/GNR systems exhibit a LS <-> HS binary behavior, which can be considered as a useful property in the development of nanomemories based upon metallic clusters. (C) 2011 American Institute of Physics. [doi:10.1063/1.3553849]

Evaluation of flow patterns and elongated bubble characteristics during the flow boiling of halocarbon refrigerants in a micro-scale channel

In the present study, quasi-diabatic two-phase flow pattern visualizations and measurements of elongated bubble velocity, frequency and length were performed. The tests were run for R134a and R245fa evaporating in a stainless steel tube with diameter of 2.32 mm, mass velocities ranging from 50 to 600 kg/m(2) s and saturation temperatures of 22 degrees C, 31 degrees C and 41 degrees C. The tube was heated by applying a direct DC current to its surface. Images from a high-speed video-camera (8000 frames/s) obtained through a transparent tube just downstream the heated sections were used to identify the following flow patterns: bubbly, elongated bubbles, churn and annular flows. The visualized flow patterns were compared against the predictions provided by Barnea et al. (1983) [1], Felcar et al. (2007) [10], Revellin and Thome (2007) [3] and Ong and Thome (2009) [11]. From this comparison, it was found that the methods proposed by Felcar et al. (2007) [10] and Ong and Thome (2009) [1] predicted relatively well the present database. Additionally, elongated bubble velocities, frequencies and lengths were determined based on the analysis of high-speed videos. Results suggested that the elongated bubble velocity depends on mass velocity, vapor quality and saturation temperature. The bubble velocity increases with increasing mass velocity and vapor quality and decreases with increasing saturation temperature. Additionally, bubble velocity was correlated as linear functions of the two-phase superficial velocity. (C) 2010 Elsevier Inc. All rights reserved.

On the accuracy of two-fluid model predictions for a particular gas-solid riser flow

Literature presents a huge number of different simulations of gas-solid flows in risers applying two-fluid modeling. In spite of that, the related quantitative accuracy issue remains mostly untouched. This state of affairs seems to be mainly a consequence of modeling shortcomings, notably regarding the lack of realistic closures. In this article predictions from a two-fluid model are compared to other published two-fluid model predictions applying the same Closures, and to experimental data. A particular matter of concern is whether the predictions are generated or not inside the statistical steady state regime that characterizes the riser flows. The present simulation was performed inside the statistical steady state regime. Time-averaged results are presented for different time-averaging intervals of 5, 10, 15 and 20 s inside the statistical steady state regime. The independence of the averaged results regarding the time-averaging interval is addressed and the results averaged over the intervals of 10 and 20 s are compared to both experiment and other two-fluid predictions. It is concluded that the two-fluid model used is still very crude, and cannot provide quantitative accurate results, at least for the particular case that was considered. (C) 2009 Elsevier Inc. All rights reserved.

Cluster identification and characterization in the riser of a circulating fluidized bed from numerical simulation results

A methodology of identification and characterization of coherent structures mostly known as clusters is applied to hydrodynamic results of numerical simulation generated for the riser of a circulating fluidized bed. The numerical simulation is performed using the MICEFLOW code, which includes the two-fluids IIT`s hydrodynamic model B. The methodology for cluster characterization that is used is based in the determination of four characteristics, related to average life time, average volumetric fraction of solid, existing time fraction and frequency of occurrence. The identification of clusters is performed by applying a criterion related to the time average value of the volumetric solid fraction. A qualitative rather than quantitative analysis is performed mainly owing to the unavailability of operational data used in the considered experiments. Concerning qualitative analysis, the simulation results are in good agreement with literature. Some quantitative comparisons between predictions and experiment were also presented to emphasize the capability of the modeling procedure regarding the analysis of macroscopic scale coherent structures. (c) 2007 Elsevier Inc. All rights reserved.

Void fraction measurement and signal analysis from multiple-electrode impedance sensors

Void fraction sensors are important instruments not only for monitoring two-phase flow, but for furnishing an important parameter for obtaining flow map pattern and two-phase flow heat transfer coefficient as well. This work presents the experimental results obtained with the analysis of two axially spaced multiple-electrode impedance sensors tested in an upward air-water two-phase flow in a vertical tube for void fraction measurements. An electronic circuit was developed for signal generation and post-treatment of each sensor signal. By phase shifting the electrodes supplying the signal, it was possible to establish a rotating electric field sweeping across the test section. The fundamental principle of using a multiple-electrode configuration is based on reducing signal sensitivity to the non-uniform cross-section void fraction distribution problem. Static calibration curves were obtained for both sensors, and dynamic signal analyses for bubbly, slug, and turbulent churn flows were carried out. Flow parameters such as Taylor bubble velocity and length were obtained by using cross-correlation techniques. As an application of the void fraction tested, vertical flow pattern identification could be established by using the probability density function technique for void fractions ranging from 0% to nearly 70%.

Multiphase flow in the vascular system of wood: From microscopic exploration to 3-D Lattice Boltzmann experiments

This paper provides insights into liquid free water dynamics in wood vessels based on Lattice Boltzmann experiments. The anatomy of real wood samples was reconstructed from systematic 3-D analyses of the vessel contours derived from successive microscopic images. This virtual vascular system was then used to supply fluid-solid boundary conditions to a two-phase Lattice Boltzmann scheme and investigate capillary invasion of this hydrophilic porous medium. Behavior of the liquid phase was strongly dependent on anatomical features, especially vessel bifurcations and reconnections. Various parameters were examined in numerical experiments with ideal vessel bifurcations, to clarify our interpretation of these features. (c) 2010 Elsevier Ltd. All rights reserved.