Association of N176K and L141F dimorphisms of the PRNP gene with lack of pathological prion protein deposition in placentas of naturally and experimentally scrapie-affected ARQ/ARQ sheep.

The placenta is important in the horizontal transmission of the aetiological agent in scrapie-affected sheep. It has been demonstrated that the placentas of fetuses carrying the dimorphism Q171R of the PRNP gene is resistant to pathological prion protein (PrP(Sc)) accumulation in the placenta. To test whether other PRNP polymorphisms are associated with a lack of placental PrP(Sc) deposition, we carried out a study on 26 naturally and 11 experimentally scrapie-affected ewes with or without clinical signs. PrP(Sc) was detected in the placenta of ARQ/ARQ(wild type) fetuses by Western blot and immunohistochemical analysis, but not in ARQN(176)/ARQK(176) or, as expected, ARQ/ARR samples. Furthermore, three of four AL(141)RQ/AF(141)RQ placentas were also PrP(Sc) negative, suggesting that the dimorphism at codon 141 may also mediate placental deposition of PrP(Sc). This finding demonstrates for the first time that fetal PRNP polymorphisms, other than those at codon 171, are associated with the lack of placental deposition of PrP(Sc).

Detection of fish allergen by droplet digital PCR.

Fish is one of fourteen allergens that must be highlighted on the label within the ingredients list. The European regulation is very restrictive to allergens with zero tolerance. Therefore, it is important to establish sensitive and specific methods for detecting fish allergen. Applicability to detect and quantify fish allergen by droplet digital polymerase chain reaction (ddPCR) has been evaluated in this work. Genomic DNA of three species belonging to the most common fish families were analyzed. PCR primers were designed to amplify a 166 bp region of the 18S rRNA gene. Comparative studies were performed to establish the optimal primer and probe concentrations. Annealing temperature was determined by using thermal gradient. The results have shown good applicability of the optimized 18S rRNA gene-method to detect and quantify small amounts of the target in samples analyzed. However, validation studies are needed in order to apply ddPCR technology for routine allergens analysis.

Antiproliferative and pro-apoptotic activity of eugenol-related biphenyls on malignant melanoma cells.

BACKGROUND: Malignant melanoma is one of the most aggressive skin cancer and chemotherapeutic agents currently in use are still unsatisfactory. Prevention and early diagnosis are the only effective tools against this tumour whose incidence and mortality rates are highly increased during the last decades in fair skin populations. Therefore the search for novel therapeutic approaches is warranted. Aim of this work was to identify and test new compounds with antiproliferative and cytotoxic activity on melanoma cells. We tested eugenol together with six natural and synthetic eugenol-related compounds for their capability to inhibit cell growth on primary melanoma cell lines established from patients' tissue samples. RESULTS: Eugenol and isoeugenol monomers and their respective O-methylated forms did not show to inhibit melanoma cells proliferation. Conversely, the dimeric forms (biphenyls) showed some antiproliferative activity which was mild for dehydrodieugenol, higher for its O,O'-methylated form (O,O'-dimethyl-dehydrodieugenol), and markedly pronounced for the racemic mixture of the brominated biphenyl (6,6'-dibromo-dehydrodieugenol) (S7), being its enantiomeric form (S) the most effective compared to the other compounds. Such activity resulted to be selective against tumour cells, without affecting cultured normal human skin fibroblasts. Dose and time dependence curves have been obtained for the enantiomeric form S7-(S). Then IC50 and minimal effective doses and times have been established for the melanoma cell lines tested. TUNEL and phosphatidylserine exposure assays demonstrated the occurrence of apoptotic events associated with the antiproliferative activity of S7-(S). Cytotoxic activity and apoptosis induced by treating melanoma cells with eugenol-related biphenyls was partially dependent by caspase activation. CONCLUSION: Our findings demonstrate that the eugenol related biphenyl (S)-6,6'-dibromo-dehydrodieugenol elicits specific antiproliferative activity on neuroectodermal tumour cells partially triggering apoptosis and its activity should be further investigated on in vivo melanoma models in order to evaluate the real anticancer effectiveness on such tumour.

Analysis of Foreign Matter in Foodstuffs Using the Light Filth Test: Report 2012-2013.

The European legislation set the new hygiene standards in food chain with the purpose to ensure high levels of public health protection in relation to food production. In order to guarantee excellent hygiene standards in food chain, particular attention must be paid to the presence of foreign matter, like light solid impurities of mineral, vegetable or animal origin. The light filth test is a suitable method used to detect and count light solid impurities applicable to different foodstuffs. We report the results of the analysis of 93 foodstuffs official samples investigated for the presence of foreign matter at the Institute for Experimental Veterinary Medicine of Sardinia, from 2012 to 2013. Insect fragments were found in a sample of semolina and in a sample of canned tomato; plastic fragments were found in a sample of grated bread.

Foreign Bodies in Dried Mushrooms Marketed in Italy.

The presence of foreign bodies in mushrooms affects their marketability and may result in health risks to consumers. The inspection of fresh or dried mushrooms today is very important in view of the increased consumption of this kind of food. Ten samples of dried mushrooms collected in supermarkets were examined for evidence of entomological contamination by macro and microscopic analytical methods, the so-called filth-test. A total of 49 46 determinations, comprising 15 g of the vegetable matrix, were made. The microscopic filth test consistently detected an irregular distribution of physical contaminants following repeated determinations of the same sample. Visual examination, on the other hand, was not sufficient to ensure a product free of contaminants.

Monitoring the Presence of Genetically Modified Potato EH92-527-1 (BPS-25271-9) in Commercial Processed Food.

The Amflora (EH92-527-1) potato is a genetically modified (GM) potato in which only starch of the amylopectin form is produced. This has been achieved by intervening with the biosynthesis of starch in this variety of potato. The Amflora potato is solely grown for the purposes of enhancing its industrial application. Although the Amflora potato is not fit for human consumption, the presence of the potato itself or any of its derived products in the food chain cannot be excluded, it should be considerate adventitious or technically unavoidable and can be accepted in a proportion no higher than 0.9%. To achieve the goal of our work we analysed forty-five potato-derived products to evaluate transgenic potato presence by real time polymerase chain reaction, obtaining negative results. In order to verify the correct application of the law and to assure the quality for the consumer, it is necessary to continue GM monitoring to verify the adventitious presence itself in food.

Microgravity alters basal and insulin-mediated metabolic activity of normal and neoplastic cells.

In this paper we report the behaviour of normal vascular smooth muscle cells and transformed breast cancer cells under normal versus simulated microgravity conditions by comparing cell proliferation, Glucose transport, Methionine uptake and protein synthesis. Modeled microgravity profoundly affects cell growth (especially in normal cells) and Glucose or Methionine metabolism (although to different extent in the two cell lines). Since both cells own responsive insulin receptors, the comparison was extended to insulin-stimulated versus unstimulated conditions. We report that the detected metabolic changes were strongly enhanced when the cells were simultaneously stimulated with insulin and subjected to modeled microgravity stress. Such observations may have important returns for human health in space; they deserve further attention.