Dynamic Solid Phase DNA Extraction and PCR Amplification in Polyester-Toner Based Microchip

A variety of substrates have been used for fabrication of microchips for DNA extraction, PCR amplification, and DNA fragment separation, including the more conventional glass and silicon as well as alternative polymer-based materials. Polyester represents one such polymer, and the laser-printing of toner onto polyester films has been shown to be effective for generating polyester-toner (PeT) microfluidic devices with channel depths on the order of tens of micrometers. Here, we describe a novel and simple process that allows for the production of multilayer, high aspect-ratio PeT microdevices with substantially larger channel depths. This innovative process utilizes a CO(2) laser to create the microchannel in polyester sheets containing a uniform layer of printed toner, and multilayer devices can easily be constructed by sandwiching the channel layer between uncoated cover sheets of polyester containing precut access holes. The process allows the fabrication of deep channels, with similar to 270 mu m, and we demonstrate the effectiveness of multilayer PeT microchips for dynamic solid phase extraction (dSPE) and PCR amplification. With the former, we found that (i) more than 65% of DNA from 0.6 mu L of blood was recovered, (ii) the resultant DNA was concentrated to greater than 3 ng/mu L., (which was better than other chip-based extraction methods), and (iii) the DNA recovered was compatible with downstream microchip-based PCR amplification. Illustrative of the compatibility of PeT microchips with the PCR process, the successful amplification of a 520 bp fragment of lambda-phage DNA in a conventional thermocycler is shown. The ability to handle the diverse chemistries associated with DNA purification and extraction is a testimony to the potential utility of PeT microchips beyond separations and presents a promising new disposable platform for genetic analysis that is low cost and easy to fabricate.

A new and improved strategy combining a dispersive-solid phase extraction-based multiclass method with ultra high pressure liquid chromatography for analysis of low molecular weight polyphenols in vegetables

This paper reports on the development and optimization of a modified Quick, Easy, Cheap Effective, Rugged and Safe (QuEChERS) based extraction technique coupled with a clean-up dispersive-solid phase extraction (dSPE) as a new, reliable and powerful strategy to enhance the extraction efficiency of free low molecular-weight polyphenols in selected species of dietary vegetables. The process involves two simple steps. First, the homogenized samples are extracted and partitioned using an organic solvent and salt solution. Then, the supernatant is further extracted and cleaned using a dSPE technique. Final clear extracts of vegetables were concentrated under vacuum to near dryness and taken up into initial mobile phase (0.1% formic acid and 20% methanol). The separation and quantification of free low molecular weight polyphenols from the vegetable extracts was achieved by ultrahigh pressure liquid chromatography (UHPLC) equipped with a phodiode array (PDA) detection system and a Trifunctional High Strength Silica capillary analytical column (HSS T3), specially designed for polar compounds. The performance of the method was assessed by studying the selectivity, linear dynamic range, the limit of detection (LOD) and limit of quantification (LOQ), precision, trueness, and matrix effects. The validation parameters of the method showed satisfactory figures of merit. Good linearity (View the MathML sourceRvalues2>0.954; (+)-catechin in carrot samples) was achieved at the studied concentration range. Reproducibility was better than 3%. Consistent recoveries of polyphenols ranging from 78.4 to 99.9% were observed when all target vegetable samples were spiked at two concentration levels, with relative standard deviations (RSDs, n = 5) lower than 2.9%. The LODs and the LOQs ranged from 0.005 μg mL−1 (trans-resveratrol, carrot) to 0.62 μg mL−1 (syringic acid, garlic) and from 0.016 μg mL−1 (trans-resveratrol, carrot) to 0.87 μg mL−1 ((+)-catechin, carrot) depending on the compound. The method was applied for studying the occurrence of free low molecular weight polyphenols in eight selected dietary vegetables (broccoli, tomato, carrot, garlic, onion, red pepper, green pepper and beetroot), providing a valuable and promising tool for food quality evaluation.

Limnology of Macrobrachium amazonicum grow-out ponds subject to high inflow of nutrient-rich water and different stocking and harvest management

We evaluated the water characteristics and particle sedimentation in Macrobrachium amazonicum (Heller 1862) grow-out ponds supplied with a high inflow of nutrient-rich water. Prawns were subject to different stocking and harvesting strategies: upper-graded juveniles, lower-graded juveniles, non-graded juveniles + selective harvesting and traditional farming (non-grading juveniles and total harvest only). Dissolved oxygen, afternoon N-ammonia and N-nitrate and soluble orthophosphate were lower in the ponds in comparison with inflow water through the rearing cycle. Ponds stocked with the upper population fraction of graded prawns showed higher turbidity, total suspended solids and total Kjeldahl nitrogen than the remaining treatments. An increase in the chemical oxygen demand:biochemical oxygen demand ratio from inlet (4.9) to pond (7.1-8.0) waters indicated a non-readily biodegradable fraction enhancement in ponds. The sedimentation mean rate ranged from 0.08 to 0.16 mm day(-1) and sediment contained >80% of organic matter. The major factors affecting pond ecosystem dynamic were the organic load (due to primary production and feed addition) and bioturbation caused by stocking larger animals. Data suggest that M. amazonicum grow-out in ponds subjected to a high inflow of nutrient-rich water produce changes in the water properties, huge accumulation of organic sediment at the pond bottom and non-readily biodegradable material in the water column. However, the water quality remains suitable for aquaculture purposes. Therefore, nutrient-rich waters, when available, may represent a source of unpaid nutrients, which may be incorporated into economically valued biomass if managed properly.

Radial Plasma Dynamic in Sequential Pinches

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Dynamic mechanical properties for polyurethane elastomers applied in elastomeric mortar

Dynamic mechanical properties of a polyurethane (PU) elastomer and a mortar processed with the same elastomer (modified polytetramethylene ether glycol (PTMEG)) were studied. The results obtained showed that the liquid aromatic amine ETHACURE (R) 300, used as cure agent, can be used to substitute the aromatic amine MOCA (R), which is usually used as cure agent in high performance elastomers. The resulting mortar produced with ETHACURE (R) 300 presents similar dynamic-mechanical thermal properties when compared with MOCA (R). However, dynamic-mechanical thermal analysis studies showed that the mortar developed with ETHACURE (R) 300 presents some advantages such as the low values of tan d, indicating a good capacity of recovery of the strain after retreating an applied force. (c) 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

Robust broadband vibration control of a flexible structure using an electrical dynamic absorber

This paper presents a simple but practical feedback control method to suppress the vibration of a flexible structure in the frequency range between 10 Hz and 1 kHz. A dynamic vibration absorber is designed for this, which has a natural frequency of 100 Hz and a normalized bandwidth (twice the damping ratio) of 9.9. The absorber is realized electrically by feeding back the structural acceleration at one position on the host structure to a collocated piezoceramic patch actuator via an analog controller consisting of a second-order lowpass filter. This absorber is equivalent to a single degree-of-freedom mechanical oscillator consisting of a serially connected mass-spring-damper system. A first-order lowpass filter is additionally used to improve stability at very high frequencies. Experiments were conducted on a free-free beam embedded with a piezoceramic patch actuator and an accelerometer at its center. It is demonstrated that the single absorber can simultaneously suppress multiple vibration modes within the control bandwidth. It is further shown that the control system is robust to slight changes in the plant. The method described can be applied to many other practical structures, after retuning the absorber parameters for the structure under control.

On high Q(2) behavior of the pion form factor for transitions gamma*gamma -> pi(0) and gamma*gamma* -> pi(0) within the nonlocal quark-pion model

The behavior of the transition pion form factor for processes gamma (*)gamma --> pi(0) and gamma (*)gamma (*) --> pi(0) at large values of space-like photon momenta is estimated within the nonlocal covariant quark-pion model. It is shown that, in general, the coefficient of the leading asymptotic term depends dynamically on the ratio of the constituent quark mass and the average virtuality of quarks in the vacuum and kinematically on the ratio of photon virtualities. The kinematic dependence of the transition form factor allows us to obtain the relation between the pion light-cone distribution amplitude and the quark-pion vertex function. The dynamic dependence indicates that the transition form factor gamma (*)gamma -->, pi(0) at high momentum transfers is very sensitive to the nonlocality size of nonperturbative fluctuations in the QCD vacuum. (C) 2000 Elsevier B.V. B.V. All rights reserved.

The Chemical Percolation Devolatilization Model Applied to the Devolatilization of Coal in High Intensity Acoustic Fields

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of temperature on the interaction between the nonionic surfactant C(12)E(5) and poly(ethylene oxide) investigated by dynamic light scattering and fluorescence methods

The interaction between the nonionic surfactant C(12)E(5) and a high molar mass (M = 5.94 x 10(5)) poly(ethylene oxide) (PEG) in aqueous solution has been examined as a function of temperature by dynamic light scattering and fluorescence methods over a broad concentration range. Clusters of small surfactant micelles form within the PEO coil, leading to its extension. The hydrodynamic radius of the complex increases strongly with temperature as well as with the concentrations of surfactant and polymer. At high concentrations of the surfactant, the coil/micellar cluster complex coexists with free C(12)E(5) micelles in the solution. Fluorescence quenching measurements show a moderate micellar growth from 155 to 203 monomers in PEO-free solutions of C(12)E(5) over a wide concentration range (0.02-2.5%) at 8 degrees C. Below 0.25% C(12)E(5), the average aggregation number (N) of the micelles is smaller in the presence of PEO than in its absence. However, N increases with increasing surfactant concentration up to a plateau value of about 270 at about 1.2% (ca. 30 mM) C(12)E(5). At high surfactant concentrations, N is larger in the presence of polymer than in its absence, a finding which is connected to a significant lowering of the clouding temperature due to the PEO at these compositions. Similar results of increasing aggregation number followed by a plateau were also found at a fixed concentration of surfactant (2.5%) and varied PEO.

Interactions between the non-ionic surfactant C(12)E(5) and poly(ethylene oxide) studied using dynamic light scattering and fluorescence quenching

Dynamic light scattering measurements have been made to elucidate changes in the coil conformation of a high molecular weight poly(ethylene oxide) (PEG) fraction when the non-ionic surfactant C(12)E(5) is present in dilute solutions. The measurements were made at 20 degrees C as functions of(a) the C(12)E(5) concentration at constant PEO concentration, (b) the PEO concentration at constant C(12)E(5) concentration, and (c) the C(12)E(5)/PEO concentration ratio. The influence of temperature on the interactions in terms of the relaxation time distributions was also examined up to the cloud point. It was found that when the C(12)E(5)/PEO weight ratio was >2 and when the temperature was >14 degrees C, the correlation functions became bimodal with well-separated components. The fast mode derives fi om individual surfactant micelles which are present in the solution at high number density. The appearance of the slow mode, which dominates the scattering, is interpreted as resulting from the formation of micellar clusters due to an excluded-volume effect when the high molar mass (M = 6 x 10(5)) PEO is added to the surfactant solution. It is shown that the micellar clusters form within the PEO coils and lead to a progressive swelling of the latter for steric reasons. The dimensions of the PEO/C(12)E(5) complex increase with increasing surfactant concentration to a value of R(H) approximate to 94 nm (R(g) approximate to 208 nm) at C-C12E5 = 3.5%. Fluorescence quenching measurements show that the average aggregation number of C(12)E(5) increases significantly on addition of the high molar mass PEG. With increasing temperature toward the cloud point the clusters increase in number density and/or become larger. The cloud point is substantially lower than that for C12E5 in water solution and is strongly dependent on the PEO concentration.

Kinetic evaluation of the pyrolysis of high density polyethylene over H-AlMCM-41 material

The AlMCM-41 material with Si/Al=50 was synthesized by hydrothermal method, using cethyltrimethylammonium as template. The protonic H-AlMCM-41 acid form was obtained by ion exchange with ammonium chloride solution and subsequent calcination. The characterization of the material by several techniques showed that a good-quality MCM-41 material was obtained. High-density polyethylene (HDPE) has been submitted to thermal degradation alone, and in presence of the exchanged H-AlMCM-41 catalyst at a concentration of 1: 1 in mass (H-AlMCM-41/HDPE). The reactor was connected on line to a gas chromatograph connected to a mass spectrometer. This process was evaluated by thermogravimetry (TG), from 350 to 600degreesC, under helium dynamic atmosphere, with heating rates of 5.0; 10.0 and 20.0 degreesC/min. From TG curves, the activation energy, calculated using a multiple heating rate integral kinetic method, decreased from 225.5 KJ.mol(-1), for the pure polymer (HDPE), to 184.7 KJ.mol(-1), in the presence of the catalyst (H-AlMCM-41/HDPE).

Synthesis and properties of branched polyethylene/high-density polyethylene blends using a homogeneous binary catalyst system composed of early and late transition metal complexes

Branched polyethylene/high-density polyethylene blends (BPE/HDPE) with a wide range of molecular weights, melt flow indexes (MFI), and intrinsic viscosity were prepared using the homogeneous binary catalyst system composed by Ni(alpha-diimine)Cl-2 (1) (alpha-diimine = 1,4-bis(2,6-diisopropylphenyl)-acenaphthenediimine) and {Tp(Ms*)} TiCl3 (2) (Tp(Ms*)=hydridobis(3-mesitylpyrazol-1-yl)(5-mesityl-pyrazol-1-yl)) activated with MAO and/or TIBA in hexane at two different polymerization temperatures (30 and 55 degreesC) and by varying the nickel loading molar fraction (x(Ni)). At all Temperatures, a non-linear correlation between the x(Ni) and the productivity was observed, suggesting the occurrence of a synergistic effect between the nickel and the titanium catalyst precursors, which is more pronounced at 55 degreesC. The molecular weight of the BPE/HDPE blends considerably decreases with increasing Al/M molar ratio. The melt flow indexes (MFI) and intrinsic viscosities (eta) are strongly affected by x(Ni), but the melting temperatures are nearly constant, 132 +/- 3 degreesC. Dynamic mechanical thermal analysis (DMTA) shows the formation of different polymeric materials where the stiffness vanes according, to the x(Ni) and temperature used in the polymerization reaction. The surface morphology of the BPE/HDPE blends studied by scanning electron microscopy (SEM) revealed a low miscibility between the PE phases resulting in the formation of a sandwich structure after etching with o-xylene.

SAE 1045 steel/WC-Co/Ni-Cu-Ni/SAE 1045 steel joints prepared by dynamic diffusion bonding: Microelectrochemical studies in 0.6 M NaCl solution

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Determination of Total Sulfur in Agricultural Samples by High-Resolution Continuum Source Flame Molecular Absorption Spectrometry

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Properties of BaBi2Ta2O9 thin films prepared by chemical solution deposition technique for dynamic random-access memory applications

We report on the properties of BaBi2Ta2O9 (BBT) thin films for dynamic random-access memory (DRAM) and integrated capacitor applications. Crystalline BBT thin films were successfully fabricated by the chemical solution deposition technique on Pt-coated Si substrates at a low annealing temperature of 650°C. The films were characterized in terms of structural, dielectric, and insulating properties. The electrical measurements were conducted on Pt/BBT/Pt capacitors. The typical measured small signal dielectric constant and dissipation factor, at 100 kHz, were 282 and 0.023, respectively, for films annealed at 700°C for 60 min. The leakage current density of the films was lower than 10-9 A/cm2 at an applied electric field of 300 kV/cm. A large storage density of 38.4 fC/μm2 was obtained at an applied electric field of 200 kV/cm. The high dielectric constant, low dielectric loss and low leakage current density suggest the suitability of BBT thin films as dielectric layer for DRAM and integrated capacitor applications.