Applicability assessment of a real-time PCR assay for the specific detection of bovine,ovine and caprine material in feedstuffs

A TaqMan real-time polymerase chain reaction (PCR) method was developed for specific detection of bovine, ovine and caprine processed animal protein (PAP) in industrial feedstuffs. The method uses species-specific primers and probes targeting short mitochondrial D-loop sequences, and a positive amplification control based on 18S rRNA gene. The applicability of the real-time PCR protocol was assessed through analysis of 126 industrial feed samples that were manufactured to reproduce rendering processes of commercial feeds destined for farmed animals. The assay successfully classified samples as positive or negative according to the ruminant composition, enabling qualitative detection of banned material in feeds at levels as low as 0.1%. Although the method provides quantitative potential, results suggest that the real quantitative capability of the assay is limited by the existing variability in terms of composition and processing treatments of the feeds, which affect the amount and quality of amplifiable DNA.     

Optimized allylamine deposition for improved pluripotential cell culture

Deposition of allylamine (ALL) by plasma enhanced chemical vapor deposition has been optimized on silicon based models. Simultaneous energy recoil detection analysis and Rutherford backscattering spectra show that 100 W deposition is ideal in terms of polymerized film formation and H content while, lower or higher power induce reduced film retention or excessive cross linking, respectively. Surface composition of the ALL film was further probed by X-ray photoelectron spectroscopy revealing a monocomponent N 1s spectrum compatible with the presence of primary amines. Optimized ALL films were applied to polycaprolactone (PCL) surfaces after Ar plasma activation with implications in the chemistry and wettability of this biocompatible polymer. Human mesenchymal stem cells (hMSCs) were cultured on ALL coated PCL surface and controls. ALL functionalized PCL was found to be especially attractive for the formation of confluent monolayers of hMSCs after 72 h of culture.     

A formulation to model the nonlinear viscoelastic properties of the vascular tissue

Nearly all soft tissues, including the vascular tissue, present a certain degree of viscoelastic material response, which becomes apparent performing multiple relaxation tests over a wide range of strain levels and plotting the resulting stress relaxation curves, nonlinear viscoelasticity of the tissue. Changes in relaxation rate at each strain may occur at multiple strain levels. A constitutive formulation considering the particular features of the vascular tissue, such as anisotropy, together with these nonlinear viscoelastic phenomena is here presented and used to fit stress?Cstretch curves from experimental relaxation tests. This constitutive model was used to fit several data set of in vitro experimental stress relaxation tests performed on ovine and porcine aorta. The good fitting of the experimental data shows the capability of the model to reproduce the viscoelastic response of the vascular tissue.     

Native and Biotechnologically Engineered Plant Proteases with Industrial Applications

Proteases occupy the most relevant position among industrial enzymes. Plant proteases have been used in medicine, detergent manufacturing, and food science for many years, but their production is diminishing in favor of those of microbial origin because lower production costs. Papain, bromelain, and ficin are the most frequently employed plant proteases, although new proteases with new and more appealing physicochemical properties for industry are still emerging. DNA technology and genetic engineering shall play, without a doubt, an important role for the production of these proteases at the industrial level. The present review focuses on the applications of traditional plant proteases as well as new proteases discovered during the last 20 years, some of which have already been genetically engineered either to increase production or to strengthen some of their physicochemical properties. The review also refers to the protease classification, action pattern, and main characteristics.     

Crystal growth induced by Nd:YAG laser irradiation in patterning glass ceramic substrates with dots

In this work a glass ceramic substrate was processed by focusing a laser beam inside the said material. The crystal phase within the amorphous matrix provides mechanical properties to the glass ceramic substrate in such a way that dots can be patterned inside the fore-mentioned material without producing any cracks. These marks are made up of crystals, the growth of which has been induced by the laser beam. These inner structures can modify the optical, thermal and mechanical properties of the glass ceramic substrate.A Q-switched Nd:YAG laser at its fundamental wavelength of 1064 nm with pulsewidths in the nanosecond range has been used.Morphology, composition, microstructure, mechanical and thermal properties of the processed material are described.     

A residual-based a posteriori error estimator for a fully-mixed formulation of the Stokes–Darcy coupled problem

In this paper we develop an a posteriori error analysis of a new fully mixed finite element method for the coupling of fluid flow with porous media flow in 2D. Flows are governed by the Stokes and Darcy equations, respectively, and the corresponding transmission conditions are given by mass conservation, balance of normal forces, and the Beavers–Joseph–Saffman law. We consider dual-mixed formulations in both media, which yields the pseudostress and the velocity in the fluid, together with the velocity and the pressure in the porous medium, and the traces of the porous media pressure and the fluid velocity on the interface, as the resulting unknowns. The set of feasible finite element subspaces includes Raviart–Thomas elements of lowest order and piecewise constants for the velocities and pressures, respectively, in both domains, together with continuous piecewise linear elements for the traces. We derive a reliable and efficient residual-based a posteriori error estimator for the coupled problem. The proof of reliability makes use of the global inf–sup condition, Helmholtz decompositions in both media, and local approximation properties of the Clément interpolant and Raviart–Thomas operator. On the other hand, inverse inequalities, the localization technique based on element-bubble and edge-bubble functions, and known results from previous works, are the main tools for proving the efficiency of the estimator. Finally, some numerical results confirming the theoretical properties of this estimator, and illustrating the capability of the corresponding adaptive algorithm to localize the singularities of the solution, are reported.     

Investigating the chlorination of acidic pharmaceuticals and by-product formation aided by an experimental design methodology

The degradation of seven acidic drugs and two metabolites during chlorination was investigated by liquid chromatography-mass spectrometry (LC-MS). A triple-quadrupole (QqQ) system was used to follow the time course of the pharmaceuticals and by-products, while a quadrupole time-of-flight (Q-TOF) system was also used for the identification of the by-products. Under strong chlorination conditions (10 mg/L Cl₂, 24 h), only four of the target compounds were significantly degraded: salicylic acid, naproxen, diclofenac and indomethacine. The degradation kinetics of these four compounds were investigated at different concentrations of chlorine, bromide and pH by means of a Box-Behnken experimental design. Depending on these factors, measured pseudo-first order half-lives were in the ranges: 23-573 h for salicylic acid, 13-446 min for naproxen, 5-328 min for diclofenac and 0.4-13.4 min for indomethacine. Also, it was observed that chlorine concentration was the overall most significant factor, followed by the bromide concentration (except for indomethacine), resulting in increased degradation kinetics as they are increased. The degradation path of salicylic acid, naproxen and diclofenac consisted of aromatic substitution of one or two hydrogens by chlorine and/or bromide. Moreover, for diclofenac, two other by-products corresponding to a decarboxylation/hydroxylation pathway from the monohalogenated products were also identified. On the other hand, indomethacine degradation did not lead to halogenation products but to oxidation ones. The investigation of these by-products in real samples by LC-MS/MS (QqQ) showed that the halogenated derivates of salicylic acid occurred in all the drinking water and wastewater samples analysed.