Validation of the performance of a GMO multiplex screening assay based on microarray detection

A new screening method for the detection and identification of GMO, based on the use of multiplex PCR followed by microarray, has been developed and is presented. The technology is based on the identification of quite ubiquitous GMO genetic target elements first amplified by PCR, followed by direct hybridisation of the amplicons on a predefined microarray (DualChip^® GMO, Eppendorf, Germany). The validation was performed within the framework of a European project (Co-Extra, contract no 007158) and in collaboration with 12 laboratories specialised in GMO detection. The present study reports the strategy and the results of an ISO complying validation of the method carried out through an inter-laboratory study. Sets of blind samples were provided consisting of DNA reference materials covering all the elements detectable by specific probes present on the array. The GMO concentrations varied from 1% down to 0.045%. In addition, a mixture of two GMO events (0.1% RRS diluted in 100% TOPAS19/2) was incorporated in the study to test the robustness of the assay in extreme conditions. Data were processed according to ISO 5725 standard. The method was evaluated with predefined performance criteria with respect to the EC CRL method acceptance criteria. The overall method performance met the acceptance criteria; in particular, the results showed that the method is suitable for the detection of the different target elements at 0.1% concentration of GMO with a 95% accuracy rate. This collaborative trial showed that the method can be considered as fit for the purpose of screening with respect to its intra- and inter-laboratory accuracy. The results demonstrated the validity of combining multiplex PCR with array detection as provided by the DualChip^® GMO (Eppendorf, Germany) for the screening of GMO. The results showed that the technology is robust, practical and suitable as a screening tool.     

Escherichia coli O157:H7 outbreak linked to raw milk cheese in Quebec, Canada: use of exact probability calculation and casecase study approaches to foodborne outbreak investigation

The analytical studies used to investigate foodborne outbreak are mostly case-control or retrospective cohort studies. However, these studies can be complex to perform and susceptible to biases. This article addresses basic principles of epidemiology, probability, and the use of case-case design to identify the source of an Escherichia coli O157:H7 outbreak linked to raw milk cheese consumption in Quebec, Canada; a small number of cases with the same pulsed-field gel electrophoresis (PFGE) profile were involved. Between 4 December 2008 and 15 January 2009, a cumulative total of 16 E. coli O157:H7 cases with the same PFGE profile were reported to Quebec public health authorities. Among the first six cases reported, three had consumed raw milk cheese from the same producer (cheese A). Raw milk cheese is consumed by about 2 % of the Quebec population. By using the exact probability calculation, it was found that a significantly higher proportion of E. coli O157:H7 cases (with the specific PFGE profile) than expected had consumed cheese A (P < 0.001). These computations were updated during the course of the investigation to include subsequent cases and gave the same results. A case-case study corroborated this result. This article considers alternative statistical and epidemiological approaches to investigate a foodborne outbreak-in particular with an exact probability calculation and case-case comparisons. This approach could offer a fast and inexpensive alternative to regular case-control studies to target public health actions, particularly during a foodborne outbreak.     

Adsorption of conditioning polymers on solid substrates with different charge density

The adsorption processes of polymers that belong to two different families (neutral hydrophilic polymers and cationic polysaccharide polymers) onto solid surfaces with different charge density have been studied using dissipative quartz crystal microbalance (D-QCM) and ellipsometry. The polymers studied are very frequently used in the cosmetic industry as conditioning agents. The adsorption kinetics of the polymers involves at least two steps. The total adsorbed amount depends on the charge density of the surface for both types of polymers. The comparison of the adsorbed mass on each layer obtained from D-QCM and from ellipsometry has allowed calculating the water content of the layers that reaches high values for the polymers studied. The analysis of D-QCM results also provided information about the shear modulus of the layers, whose values have been found to be typical of a rubber-like polymer system. The main driving force of the adsorption was found to be the energy of the interactions between chains and surface.     

Transmission Electron Microscopy Investigation of the Oxidation of Hot Isostatically Pressed Silicon Nitride with and without Sintering Aids

Analytical transmission electron microscopy of thin-foil cross sections has been used to examine the oxidation behavior of hot isostatically pressed silicon nitride (Si₃N₄) materials. The transmission electron microscopy (TEM) cross sections are prepared by a special technique that provides electron transparency through the entire oxide, interfacial, subscalar, and matrix regions simultaneously. The materials are oxidized in an alumina furnace at 1250°C for 100 h. TEM investigation indicates that oxidation of Si₃N₄ occurs in an oxidation reaction zone that is comprised of the scale, oxide/matrix interface, and subscalar regions; therefore, the silica (SiO₂)/Si₃N₄ interfacial surface area that is available for oxidation is very large. The oxidative attack on the Si₃N₄ grains is not uniform or sequential, and oxygen diffuses into the matrix before the surface grains are consumed. Gas bubbles, probably nitrogen gas, accumulate at all levels of the scale, and no evidence is found for the existence of an "oxynitride" layer. Disintegration of the secondary phase, Y₂Si₂O₇, in the subscalar region is observed to occur, indicating that secondary, oxidation-related phenomena are occurring.     

Multilayer films ageing under ultraviolet radiations: Complementary study by dielectric spectroscopy

The effect of ultraviolet (UV) irradiation on a multilayer film made of poly(ethylene terephthalate)/Polyamide 6/poly(ethylene terephthalate) was investigated by uniaxial tractions, UV‐visible‐NIR and Fourier transformed infra‐red‐attenuated total reflection spectroscopy, differential scanning calorimetry ( DSC), and dynamic dielectric spectroscopy (DDS). The multilayer was exposed to ultraviolet radiations (filtered at 270 nm) for 7 days, in air. The complexity of the multilayer thermograms recorded by DSC and DDS has required the study of each film constituting the multilayer to assess each the contribution of each one of them. A deterioration in mechanical properties and a decrease in UV transmission for low wavelengths are observed. These evolutions seem to result to the photo‐oxidation of the poly(ethylene terephthalate) film mainly localized at the exposed layer. This layer acts as a UV protection filter for the other layers. However, the DDS analyses show a plasticization effect of the primary mode in the Polyamide 6, which is evidence of photo‐oxidation. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

Pentachlorophenylcellulose preparation and investigation on its applicability as a controlled release fungicide

Pentachlorophenylethers (PCP-ethers) of cellulose are prepared using cellulosetosylates as intermediates. The products release no measurable quantities of PCP into buffered solutions (pH 4, 7, and 9, resp.). However, the modified cellulose shows no fungicidal effect in agar plate tests with the usually cellulose-metabolizing fungus, Trichoderma viride. The lack of biological activity is attributed to the stability of the cellulose pentachlorophenyl ether bond.     

GIS-based applications of sensitivity analysis for sewer models

Sensitivity analysis (SA) evaluates the impact of changes in model parameters on model predictions. Such an analysis is commonly used when developing or applying environmental models to improve the understanding of underlying system behaviours and the impact and interactions of model parameters. The novelty of this paper is a geo-referenced visualization of sensitivity indices for model parameters in a combined sewer model using geographic information system (GIS) software. The result is a collection of maps for each analysis, where sensitivity indices (calculated for model parameters of interest) are illustrated according to a predefined symbology. In this paper, four types of maps (an uncertainty map, calibration map, vulnerability map, and design map) are created for an example case study. This article highlights the advantages and limitations of GIS-based SA of sewer models. The conclusion shows that for all analyzed applications, GIS-based SA is useful for analyzing, discussing and interpreting the model parameter sensitivity and its spatial dimension. The method can lead to a comprehensive view of the sewer system.     

Dielectric relaxations and ferroelectric behaviour of even-odd polyamide PA 6,9

Thermo Stimulated Current (TSC) combined with Dynamic Dielectric Spectroscopy (DDS) have been applied to the investigation of dielectric relaxation modes of an even-odd Polyamide PA 6,9. The correlation between results obtained by both methods allows us to describe precisely the molecular mobility. At high temperature, the various dielectric relaxation phenomena are separated by applying the dielectric modulus formalism. The comparison between the activation enthalpy values obtained by DDS and TSC leads to the assignment of the so-called α mode to cooperative movements of polymeric sequences. Molecular mobility of PA 6,9 is compared with the one of PA 11. The piezoelectric activity of PA 6,9 is shown and analyzed.     

Dynamic mechanical behaviour of polyamide 11/Barium titanate ferroelectric composites

Dynamic mechanical analysis and tensile test have been used to characterize the mechanical behaviour of hybrid composites. Barium titanate (BaTiO₃) is the submicron filler and polyamide 11 (PA 11) the matrix. The influence of volume fraction and particles size (ranging from 100 nm to 700 nm) of the inorganic phase on the composites mechanical properties have been checked. BaTiO₃ dispersion in the matrix increases the tensile modulus of the composites and an evolution from ductile to fragile is observed for volume fractions above 12 vol%. The volume fraction dependence of the glassy shear modulus is well described by the Hashin and Shtrikman model indicative of an interaction lack between the organic and inorganic phases. The decrease of the glassy shear modulus with the filler size has been associated with the existence of softer organic/inorganic interfaces, in agreement with the previous hypothesis. The non linear variation of the rubbery modulus versus particles content is well described by the rubber elasticity model applied to a hydrogen bond network.