Application of the novel Droplet digital PCR technology for identification of meat species

The authenticity and traceability of meat products are issues of primary importance to ensure food safety. Unfortunately, food adulteration (e.g. the addition of inexpensive cuts to minced meat products) and mislabelling (e.g. the inclusion of meat from species other than those declared) happens frequently worldwide. The aim of this study was to apply a droplet digital PCR assay for the detection and quantification (copies μL⁻¹) of the beef, pork, horse, sheep, chicken and turkey in meat products. The analysis conducted on commercial meat showed the presence of traces of DNA from other animal species than those declared. We show that the method is highly sensitive, specific and accurate (accuracy = 100%). This method could be adopted by competent food safety authorities to verify compliance with the labelling of meat products and to ensure quality and safety throughout the meat supply chain, from primary production to consumption.     

Lc/Esi/Ms/Ms determination of postharvest fungicide residues in citrus juices

An LC-ESI-MS/MS method was developed for the quantitative detection of postharvest fungicide residues in citrus juices and reported in this paper. The analyses of thiabendazole (TBZ), carbendazim (MBC), thiophanate methyl (TPM), imazalil (IMZ) and prochloraz (PCZ) residues were performed by using a gradient elution in conjunction with positive ionization mode electrospray ionization tandem mass spectrometry. Fungicides were extracted from citrus juices with recoveries ranging from 79.8 to 101.2% and relative standard deviation better than 15%. The quantification limits ranged from 0.01 μg/kg IMZ to 0.06 μg/kg for MBC. The LC-MS-MS method was applied to commercial citrus juices, detecting MBC, TBZ and IMZ residues in the 90% of the samples. Prochloraz residues were detected only in one of the multifruit juice (orange, lemon and carrot) samples.     

Preparation and physico-chemical characterisation of microporous polysaccharidic hydrogels

A new technique to obtain microporous hydrogels was realised. It permits us to obtain a microporous structure directly on the already cross-linked hydrogel. It consists in stratifying the already cross-linked hydrogel on to a filter with known porosity and forcing the CO2 bubbles, derived from the addition of HCl to a porogen salt (NaHCO3), to cross through the filter first and then the matrix. By changing the porosity of the filter, it was possible to modulate the porous morphology of the hydrogels. The polysaccharides selected were hyaluronane, alginate, and carboxymethylcellulose. The influence of the porous morphology on the physico-chemical properties of the gel has been evaluated by FT-IR, FRAP, calorimetric, water uptake, and rheological analysis.     

Formulative Study and Intracellular Fate Evaluation of Ethosomes and Transethosomes for Vitamin D3 Delivery

In this pilot study, ethosomes and transethosomes were investigated as potential delivery systems for cholecalciferol (vitamin D3), whose deficiency has been correlated to many disorders such as dermatological diseases, systemic infections, cancer and sarcopenia. A formulative study on the influence of pharmaceutically acceptable ionic and non-ionic surfactants allowed the preparation of different transethosomes. In vitro cytotoxicity was evaluated in different cell types representative of epithelial, connective and muscle tissue. Then, the selected nanocarriers were further investigated at light and transmission electron microscopy to evaluate their uptake and intracellular fate. Both ethosomes and transethosomes proven to have physicochemical properties optimal for transdermal penetration and efficient vitamin D3 loading; moreover, nanocarriers were easily internalized by all cell types, although they followed distinct intracellular fates: ethosomes persisted for long times inside the cytoplasm, without inducing subcellular alteration, while transethosomes underwent rapid degradation giving rise to an intracellular accumulation of lipids. These basic results provide a solid scientific background to in vivo investigations aimed at exploring the efficacy of vitamin D3 transdermal administration in different experimental and pathological conditions.     

Antioxidant activity of tomato lipophilic extracts and interactions between carotenoids and α-tocopherol in synthetic mixtures

In the present study we assayed the antioxidant activity of lipophilic extracts obtained from different tomato varieties. The results showed that cherry tomatoes, characterized by a high carotenoid content, had the highest antioxidant activity. A quantitative analysis of lycopene, β-carotene, lutein and α-tocopherol was also performed and the correlation between the antioxidant content and the antioxidant activity was estimated. The highest correlation coefficient was found for lycopene (R² = 0.9236, P ≤ 0.001). The analysis of two-component mixtures containing α-tocopherol and carotenoids showed that significant synergism occurred for all the combinations which contained α-tocopherol and β-carotene mixed together. The highest synergistic effects were detected for α-tocopherol-lycopene mixtures, which were the most efficient combinations tested in the present study. The analysis of the carotenoid combinations indicated that synergism occurred for lycopene-β-carotene, lycopene-lutein and lutein-β-carotene mixtures. The analysis of four-component mixtures did not show statistically significant synergistic effects.     

Transposable Elements and Stress in Vertebrates: An Overview

Since their identification as genomic regulatory elements, Transposable Elements (TEs) were considered, at first, molecular parasites and later as an important source of genetic diversity and regulatory innovations. In vertebrates in particular, TEs have been recognized as playing an important role in major evolutionary transitions and biodiversity. Moreover, in the last decade, a significant number of papers has been published highlighting a correlation between TE activity and exposition to environmental stresses and dietary factors. In this review we present an overview of the impact of TEs in vertebrate genomes, report the silencing mechanisms adopted by host genomes to regulate TE activity, and finally we explore the effects of environmental and dietary factor exposures on TE activity in mammals, which is the most studied group among vertebrates. The studies here reported evidence that several factors can induce changes in the epigenetic status of TEs and silencing mechanisms leading to their activation with consequent effects on the host genome. The study of TE can represent a future challenge for research for developing effective markers able to detect precocious epigenetic changes and prevent human diseases.