SYBR®Green qPCR screening methods for the presence of “35S promoter” and “NOS terminator” elements in food and feed products

The Cauliflower Mosaic Virus “35S promotor” (p35S) and the Agrobacterium “Nopaline Synthase” terminator (tNOS) are the most represented generic recombinant elements in commercial genetically modified crops to date. A set of four new SYBR^®Green qPCR methods targeting the “p35S” and “tNOS” core elements have been developed. These qPCR methods generate short amplicons of 147 and 75 bp for the “p35S” element and 172 and 69 bp for the “tNOS” element. Single target plasmids containing these amplicons were constructed and allow determining the nominal melting temperature (T _m value) of each amplicon. The four methods are specific for their respective targets, and moreover, three of them are highly sensitive (up to 1–2 copies detectable) at a PCR efficiency ranging between 95 and 100%. The latter methods can detect their respective targets at 0.1% (w/w) gDNA levels and are suitable for detecting low levels of genetically modified materials containing the “p35S” and/or “tNOS” elements.     

Extreme catalysts from low-temperature environments

Cold-loving or psychrophilic organisms are widely distributed in nature as a large part of the earth's surface is at temperatures around 0 degrees C. To maintain metabolic rates and to prosper in cold environments, these extremophilic organisms have developed a vast array of adaptations. One main adaptive strategy developed in order to cope with the reduction of chemical reaction rates induced by low temperatures is the synthesis of cold-adapted or psychrophilic enzymes. These enzymes are characterized by a high catalytic activity at low temperatures associated with a low thermal stability. A study of protein adaptation strategies suggests that the high activity of psychrophilic enzymes could be achieved by the destabilization of the active site, allowing the catalytic center to be more flexible at low temperatures, whereas other protein regions may be destabilized or as rigid as their mesophilic counterparts. Due to these particular properties, psychrophilic enzymes offer a high potential not only for fundamental research but also for biotechnological applications.     

Freshness characterisation of whiting (Merlangius merlangus) using an SPME/GC/MS method and a statistical multivariate approach

BACKGROUND: The freshness of whiting was studied at five stages of ice storage by comparing the analysis of volatile compounds obtained through solid phase microextraction/gas chromatography/mass spectrometry (SPME/GC/MS) with two sensory methods. RESULTS: Of the volatile compounds identified, 38 were analysed using a statistical multivariate approach and classified according to their role in the estimation of freshness during storage as markers of freshness or spoilage. Regarding the evolution of the presence or absence of individual compounds, three categories were defined. For example, the volatile compounds propanal, hexanal, 1‐penten‐3‐ol, pentanal, 2,3‐pentanedione, 1‐penten‐3‐one, heptanal, (E)‐2‐pentenal, 2,3‐octanedione, (Z)‐2‐penten‐1‐ol, 1‐pentanol, butanal, octanal, 3,5,5‐trimethyl‐2‐hexene, 1‐hexanol and 4,4‐dimethyl‐1,3‐dioxane appeared highly relevant, because they are found throughout storage and can be divided into several categories that are directly related to the quality of fish. CONCLUSION: SPME/GC/MS combined with a statistical multivariate approach may be a useful method to identify volatile compounds and characterise fish freshness during storage. Copyright © 2010 Society of Chemical Industry     

Wheat gluten-coated papers for bio-based food packaging: Structure,surface and transfer properties

The objective of the present work was to understand how the structural, surface, water vapour and gas barrier properties of wheat gluten (WG) coated paper could be influenced by the features of paper. For this purpose, a surface treated paper (TP) and an untreated paper (UTP) were compared. Penetration of WG-coating into the bulk of paper was more pronounced in UTP than TP. This led to the formation of a significant junction zone resulting in an interpenetrated structure for WG–UTP, suggesting a composite-like structure, while a bi-layer one was obtained for WG–TP. Differences in WG penetration were related to the hydrophilicity, roughness and porosity of the paper used. Even though the extent of penetration did not greatly affect the surface properties (water and oil resistance), the transfer properties (water vapour, O₂ and CO₂) of WG-coated papers were significantly improved when WG-coating highly penetrated: while the WG–TP behaved as a micro-perforated material, the WG–UTP behaved as WG-film.     

Influence of pressurised cryogenic nitrogen technology on Arthrospira platensis (spirulina) preservation during storage

In this study, preservation of spirulina using the new pressurised cryogenic nitrogen technology (PCN) was compared to classical methods used in laboratories and industry. Spirulina morphology was better preserved by PCN compared to unpressurised cryogeny and classical freezing at −20 °C that led to cells fragmentation. A 25% loss of Phycocyanin-C content against 60% were measured after 98 days storage for 6-Bar PCN process and frozen samples, respectively. The Total AntiOxidant Power (PAOT Liquid Technology^®) was used for determination of total antioxidant and oxidant power of spirulina extracts. PAOT value of PCN samples was 50% higher than the frozen sample. From ABTS measurements on PCN spirulina fractions sonicated or not, it was suggested that pressurisation at 6 bars allowed a better preservation of free antioxidants (outside the cells) due to replacement of oxygen by nitrogen in the frozen beads. After dehydration, phycocyanin-C content variation during storage at 20 °C and 33% RH showed higher loss for freeze-dried spirulina treated at 0 compared to 6 Bars.     

Distinctive arsenic(V) trapping modes by magnetite nanoparticles induced by different sorption processes

Arsenic sorption onto iron oxide spinels such as magnetite may contribute to arsenic immobilization at redox fronts in soils, sediments, and aquifers, as well as in putative remediation and water treatment technologies. We have investigated As(V) speciation resulting from different sorption processes on magnetite nanoparticles, including both adsorption and precipitation, using X-ray absorption fine structure (XAFS) spectroscopy and transmission electron microscopy (TEM). XAFS results suggest that AsO(4) tetrahedra form predominantly inner-sphere bidentate corner-sharing ((2)C) complexes and outer-sphere complexes on magnetite in the adsorption experiments. In the precipitation experiments, an increasing fraction of AsO(4) tetrahedra appears to be incorporated in clusters having a magnetite-like local structure with increasing As loading, the remaining fraction of As being adsorbed at the surface of magnetite particles. In the sample with the highest As loading (15.7 μmol/m(2)) XAFS data indicate that As(V) is fully incorporated in such clusters. Such processes help to explain the significantly higher arsenic uptake in precipitation samples compared to those generated in adsorption experiments. In addition, for the precipitation samples, TEM observations indicate the formation of amorphous coatings and small (~3 nm) nanoparticles associated with larger (~20-40 nm) magnetite nanoparticles, which are absent in the adsorption samples. These results suggest that As(V) could form complexes at the surfaces of the small nanoparticles and could be progressively incorporated in their structure with increasing As loading. These results provide some of the fundamental knowledge about As(V)-magnetite interactions that is essential for developing effective water treatment technologies for arsenic.     

Bacterial formation of tooeleite and mixed arsenic(III) or arsenic(V)-iron(III) gels in the Carnoulès acid mine drainage,France. A XANES,XRD, and SEM study

The oxidation of Fe(II) in acid mine drainage (AMD) leads to the precipitation of Fe(III) compounds which may incorporate toxic elements, such as arsenic (As), within their structure or adsorb them at their surface, thus limiting their mobility. The present work provides evidence for spatial and seasonal variations of microbial activity that influence arsenite oxidation and As immobilization in the heavily contaminated AMD from the Carnoulès mine, Gard, France ([As III] = 80 to 280 mg x L(-1) in the acidic spring draining the waste-pile). In the first tens of meters of the AMD, the rapid oxidation of Fe(II) leads to the coprecipitation of large amounts of As with Fe(III) in bacterial mats. XRD, XANES, and SEM analyses of sediments and stromatolite samples revealed the unusual formation of As(III)-rich compounds, especially nanocrystalline tooeleite, Fe6(AsO3)4(SO4)(OH)4 x 4H2O, a rare ferric arsenite sulfate oxy-hydroxide mineral, together with XRD-amorphous mixed As(III)/As(V)-Fe(III) oxy-hydroxide compounds. In the wet season, the suspended sediments of the upstream zone essentially consist of tooeleite associated with am-As(III)-Fe(III) oxy-hydroxides, while am-As(V)-Fe(III) oxy-hydroxides, having As:Fe molar ratios as high as 0.6-0.8, dominate in the dry season. Comparing natural and bioassay samples revealed that the formation of As(III)-rich compounds in the wet season may be related to the metabolic activity of bacterial strains able to oxidize Fe(II) but not As(III). One of these strains, having an Acidithiobacillus ferrooxidans genotype, has been isolated from the Carnoulès AMD. In contrast, the formation of As(V)-rich compounds in the dry season can be related to both biotic and abiotic oxidation of As(III) to As(V). Some Thiomonas strains isolated from the Carnoulès AMD were shown to be able to catalyze the oxidation of As(III) to As(V) in solution. Therefore, they can promote the formation of mixed As(V)-Fe(III) oxy-hydroxides, provided enough Fe(II) oxidizes. These results yield a better understanding of natural processes at this site and may help in designing efficient As-removal processes.     

Effect of imputing markers from a low-density chip on the reliability of genomic breeding values in Holstein populations

The purpose of this study was to investigate the imputation error and loss of reliability of direct genomic values (DGV) or genomically enhanced breeding values (GEBV) when using genotypes imputed from a 3,000-marker single nucleotide polymorphism (SNP) panel to a 50,000-marker SNP panel. Data consisted of genotypes of 15,966 European Holstein bulls from the combined EuroGenomics reference population. Genotypes with the low-density chip were created by erasing markers from 50,000-marker data. The studies were performed in the Nordic countries (Denmark, Finland, and Sweden) using a BLUP model for prediction of DGV and in France using a genomic marker-assisted selection approach for prediction of GEBV. Imputation in both studies was done using a combination of the DAGPHASE 1.1 and Beagle 2.1.3 software. Traits considered were protein yield, fertility, somatic cell count, and udder depth. Imputation of missing markers and prediction of breeding values were performed using 2 different reference populations in each country: either a national reference population or a combined EuroGenomics reference population. Validation for accuracy of imputation and genomic prediction was done based on national test data. Mean imputation error rates when using national reference animals was 5.5 and 3.9% in the Nordic countries and France, respectively, whereas imputation based on the EuroGenomics reference data set gave mean error rates of 4.0 and 2.1%, respectively. Prediction of GEBV based on genotypes imputed with a national reference data set gave an absolute loss of 0.05 in mean reliability of GEBV in the French study, whereas a loss of 0.03 was obtained for reliability of DGV in the Nordic study. When genotypes were imputed using the EuroGenomics reference, a loss of 0.02 in mean reliability of GEBV was detected in the French study, and a loss of 0.06 was observed for the mean reliability of DGV in the Nordic study. Consequently, the reliability of DGV using the imputed SNP data was 0.38 based on national reference data, and 0.48 based on EuroGenomics reference data in the Nordic validation, and the reliability of GEBV using the imputed SNP data was 0.41 based on national reference data, and 0.44 based on EuroGenomics reference data in the French validation.     

AA7075高强铝合金热冲压流变行为本构模型对比研究

在热冲压过程中,AA7075高强铝合金板料经充分固溶后移入室温模具进行冲压成形并淬火。为表征AA7075铝合金在热冲压工艺中的变形行为,在温度200~480℃、应变速率0.01~10s-1范围内进行了高温拉伸试验。基于Arrhenius类型本构模型、Johnson-Cook模型以及Zerilli-Armstrong模型提出了多种修正本构模型,并应用实验所获流变曲线进行了拟合。提出的修正模型通过将模型参数表示为应变、应变速率及温度相关的多项式函数耦合了应变、应变速率及温度对流变应力的影响,并通过均方误差(MSE)以及相关系数R值对模型流变应力预测准确性进行了评价。结果表明,修正的Johnson-Cook模型能够更加准确的预测AA7075高温流变行为。