A quadruplex PCR (qxPCR) assay for adulteration in dairy products.

This study describes the development of a quadruplex quantitative Real Time PCR (qxPCR) based on SYBR®GreenER chemistry, for rapid identification of DNA of cow, goat, sheep and buffalo in dairy products, and for quantification of cow DNA in these products. The platform was applied to: (i) mixes of milks at fixed percentages; (ii) cheeses prepared with the same mixes; (iii) commercial dairy products. The methodology enabled the detection of DNA from cow in mixes of milk and cheeses with a limit of detection (LOD) of 0.1%. When applied to commercial dairy products the qxPCR gave results comparable with each single-plex Real Time PCR. A good correlation (R(2)>0.9) between peaks' area of derivative of melting curves of amplicons and percentages of cow milk in milk mixes and cheeses, allows for an estimation of cow DNA in a dynamic range varying from 0.1-5% to 1-25%.

Comparison of protein variations in Thlaspi caerulescens populations from metalliferous and non-metalliferous soils.

In this work we analysed the protein variations which occurred in two Thlaspi caerulescens populations when subjected to 0 and 10 microM nickel (Ni) treatments: the Ni hyperaccumulator T. caerulescensfrom a metalliferous soil in Italy and T. caerulescens from Czech Republic, adapted to grow on a non-metalliferous soil. Ni accumulation in roots and shoots and the effect on growth and morphology were examined. Leaves proteins profiles of Ni treated and untreated samples were analysed by two dimensional liquid chromatography technique. From the comparison of more than 500 proteins, few differences were observed between treated and untreated plants of the same population. Differences were found between the two Thlaspi populations, instead. Proteins involved in transport, metal chelation, and signal transduction increased in abundance in the 10 microM Ni treated samples while, in condition of absence of Ni, proteins involved in sulphur metabolism, protection against reactive oxygen species and stress response showed to increase in abundance in the two populations. These proteins can be used as biomarkers both for monitoring biodiversity in indigenous plants and for selection of Ni phytoremediation plants.

Environmental risk assessment in GMO analysis.

Genetically modified or engineered organisms (GMOs, GEOs) are utilised in agriculture, expressing traits of interest, such as insect or herbicide resistance. Soybean, maize, cotton and oilseed rape are the GM crops with the largest acreage in the world. The distribution of GM acreage in the different countries is related with the different positions concerning labelling of GMO products: based on the principle of substantial equivalence, or rather based on the precautionary principle. The paper provides an overview on how the risks associated with release of GMO in the environments can be analysed and predicted, in view of a possible coexistence of GM and non-GM organisms in agriculture.Risk assessment procedures, both qualitative and quantitative, are compared in the context of application to GMOs considering also legislation requirements (Directive 2001/18/EC). Criteria and measurable properties to assess harm for human health and environmental safety are listed, and the possible consequences are evaluated in terms of significance.Finally, a mapping of the possible risks deriving from GMO release is reported, focusing on gene transfer to related species, horizontal gene transfer, direct and indirect effects on non target organisms, development of resistance in target organisms, and effects on biodiversity.

Environmental risk assessment in GMO analysis.

Genetically modified or engineered organisms (GMOs, GEOs) are utilised in agriculture, expressing traits of interest, such as insect or herbicide resistance. Soybean, maize, cotton and oilseed rape are the GM crops with the largest acreage in the world. The distribution of GM acreage in the different countries is related with the different positions concerning labelling of GMO products: based on the principle of substantial equivalence, or rather based on the precautionary principle. The paper provides an overview on how the risks associated with release of GMO in the environments can be analysed and predicted, in view of a possible coexistence of GM and non-GM organisms in agriculture.Risk assessment procedures, both qualitative and quantitative, are compared in the context of application to GMOs considering also legislation requirements (Directive 2001/18/EC). Criteria and measurable properties to assess harm for human health and environmental safety are listed, and the possible consequences are evaluated in terms of significance.Finally, a mapping of the possible risks deriving from GMO release is reported, focusing on gene transfer to related species, horizontal gene transfer, direct and indirect effects on non target organisms, development of resistance in target organisms, and effects on biodiversity.

A 2-D liquid-phase chromatography for proteomic analysis in plant tissues.

Two-dimensional liquid chromatography based on a high-performance chromatofocusing in the first dimension followed by high-resolution reversed-phase chromatography in the second dimension can be used as a complementary approach to protein separation with two-dimensional gel electrophoresis. In this work, Arabidopsis thaliana proteins obtained from different tissue extracts were resolved by using a new automated system, ProteomeLab PF 2D commercialized by Beckman Coulter (Fullerton, CA, USA). In particular, protein patterns obtained after two different extraction procedures (MgSO4 and urea buffer) were compared. Reproducibility of the protein patterns was also confirmed in different injections of the same sample and in the comparative analyses of some proteins by MALDI-TOF/MS. Computer analysis of the chromatograms revealed that with this two-dimensional liquid phase technique, hundreds of "virtual bands" can be identified and compared in crude plant protein lysates.