Tomato ionomic approach for food fortification and safety.

Food fortification is an issue of paramount of importance for people living both in developed and in developing countries. Among substances listed as "nutriceuticals", essential minerals have been recognised for their involvement in several healthy issues, involving all ages. In this frame, food plants are playing a pivotal role since their capability to compartmentalise ions and proteinmetal complexes in edible organs. Conversely, the accumulation of high metal levels in those organs may lead to safety problems. In the recent years, thanks to the availability of new and improved analytical apparatus in both ionic and genomic/transcrittomics areas, it is became feasible to couple data coming from plant physiology and genetics. Ionomics is the discipline that studies the cross-analysis of both data sets. Our group, in the frame of GenoPom project granted by MiUR, is interested to study the ionomics of tomatoes cultivars derived by breeding programmes in which wild relatives have been used to transfer several useful traits, such as resistance to biotic or abiotic stresses, fruit composition and textiture, etc. The introgression of the wild genome into the cultivated one produces new gene combinations. They might lead to the expression of some traits, such as increased or reduced adsorption of some metals and their exclusion or loading into edible organs, thus strongly involving the nutritional food value. Our final goal is to put together data coming from ions homeostasis and gene expression analyses, thus obtaining an ionomic tomato map related to ions absorption, translocation and accumulation in various plant organs, fruits included. To follow our hypothesis, we are studying the ionome of Solanum lycopersicum cv. M82 along with 76 Introgression Lines (ILs) produced by interspecific crosses between this cultivar and the wild species S. pennellii. These ILs are homozygous for small portions of the wild species genome introgressed into the domesticated M82 one. They are used as a useful tool for mapping QTL associated with many traits of interest. It is worthy to note that, until now, little information is available on QTL for ions accumulation in tomato. Moreover, as our knowledge, effects of new gene combinations in introgressed lines on ions uptake related to food safety have not been extensively studied. In this presentation we show results coming from the ionome analysis, carried out on S . lycopersicum M82 and several ILs. Plants were grown in pots in a greenhouse and watered with deionised water Thirty day-old plants were left to grow for 15 days in the presence of non-toxic concentration of Cd, Pb, As, Cr and Zn given combined. Leaves of all plants were then harvested and stored at -80°C for ionome and gene expression analyses. Preliminary results of ionome analysis of S. lycopersicum M82 and several ILs, carried out using an ICP-MS, showed that traits correlated to toxic metals and micronutrients accumulation in apical leaves were significantly modified in response to specific genetic backgrounds. Those results are perhaps due to the introgression of traits linked to uptake, translocation and accumulation of useful and/or toxic metal into plant apical leaves and to interactions of the wild type introgressed genomic regions with the cultivated genome. Also, data are shown on the identification and isolation of Solanum gene sequences related to ions uptake, translocation and accumulation, useful for further real-time gene expression evaluation in both cultivated and ILs during the treatments with the above-mentioned metals

Evaluation of degree of conversion, rate of cure, microhardness, depth of cure and contraction stress of three nano hybrid composites containing pre-polymerized spherical filler

Aim: Manufacturers aim at improving filler technology to enhance the properties of the restorative materials, thus maximising the aesthetic and functional outcome of the restored tooth. The present study tested the degree of conversion (DC), rate of cure (RC), microhardness (VHN), depth of cure (VHR) and contraction stress (CS) of three new nano hybrid composites with pre-polymerized spherical filler. Methods: Three commercially available composite resin were characterised in the present study, namely the Ceram.X\uae universal shade A3 (CXUA3), Ceram.X\uae duo enamel shade E2, and Ceram.X\uae duo dentin shade D3 (CXDE2 and CXDD3). The materials were light-cured with a LED light (SmartLite Focus, measured output 1301 mW/cm2) following the protocol recommended by the manufacturer. DC was assessed by means of Fourier-transform infrared spectroscopy, calculating RC from a second-grade polynomial fitting of the kinetic curve. A microhardness testing machine equipped with a Vickers indenter served to measure the top and bottom VHN of 2 mm-high disc-shaped specimens, using the bottom/top surface values ratio (VHR) as indirect evaluation of the depth of cure. CS vs time was evaluated by a universal testing machine provided with an extensometer as feedback system, CS was normalized for the specimen bonding area. All data sets underwent statistical analysis with dedicated software and tested for the assumptions for the use of parametric tests. Multiple analyses of variance with Scheff\ue9 post hoc test were carried out to compare the dependent variables of interest among the tested materials. Results: All tested materials exhibited a DC lower than 50%, with CXUA3 reaching the lowest DC value after 10 s. RC of CXUA3 at 5 s was comparable to that of CXDE2, while after 10s RC of CXUA3 decreased to a value proportional to that of CXDD3. For all the tested materials, top-VHN was greater than bottom-VHN. Top-VHN of CXDE2 was lower than CXUA3 and CXDD3. CXDD3 was the only material achieving VHR>80%. The main differences in CS among the tested materials were found during the irradiation with curing-light: CXDE2 displaying the lowest CS after 10 s and CXDD3 the highest after 30 s. Conclusion: The present study proved that the light curing protocol suggested by the manufacturer for the three composites might be improved: 10 s of irradiation seemed insufficient to adequately cure CXUA3 and CXDE2. Longer curing times for these materials appear advisable

In vitro and in vivo evaluation of T and B lymphocyte functions in AKR mice.

To investigate whether AKR spontaneous leukaemogenesis is associated with a reduction in functional activity of T lymphocytes, the PHA response of AKR blood cells at different ages up to and including the preleukaemic period was studied. No significant differences were observed among young, adult and preleukaemic donors. In addition, the in vitro and in vivo AKR lymphocyte functions were compared with those of CBA lymphocytes by means of their response to stimulation with T and B lymphocyte selective mitogens (PHA, Con A and LSP respectively), and their response to immunization with thymus dependent (SRBC) or independent (LPS) antigens. We observed in vitro that while the B lymphocytes responded normally to mitogen, an intrinsic hyporeactivity to mitogens characterizes the T lymphocytes. Moreover, AKR mice exhibited a reduced in vivo response to both thymus dependent and independent antigens

Chronic pelvic pain: comorbidity between chronic musculoskeletal pain and vulvodynia.

Chronic pelvic pain (CPP) is a common condition that has a major impact on the quality of life of both men and women. Male CPP is usually attributable to well-defined urogenital conditions (most frequently infectious/non infectious prostatic diseases) or musculoskeletal or bowel diseases, whereas the features of female CPP are much more complex and are of particular clinical and epidemiological importance. It is a multifactorial syndrome that can be due to diseases of the urogenital, gastrointestinal, or musculoskeletal systems, or to neurological or neuropsychiatric disorders. It is not always easy to identify its predominant pathogenesis, although it often occurs as a central sensitization syndrome triggered by an initial stimulus which is no longer detectable and only manifests itself clinically through pain. In this respect, there are some very interesting relationships between vulvodynia and fibromyalgic syndrome, as identified in a preliminary study of women with chronic musculoskeletal pain in which it was demonstrated that vulvar pain plays an important role, although it is often overlooked and undiagnosed

In vitro evidence for CCl4 metabolites covalently bound to lipoprotein micelles

AbstractCCl4-induced impairment of the lipoprotein secretion pathway of intact rat hepatocytes was carried out using 14CCl4 to check the possibility of binding to lipoproteins by CCl4 metabolites. After separation of different cell suspension fractions by means of ultracentrifugation and chemical precipitation procedures, a significant amount of the radioisotope was found covalently bound to the lipid and protein components of low density lipoproteins. Suitable experiments demonstrated that the bound radioisotope was represented by CCl4 metabolites and not by unactivated CCl4

Measurement of the parameter kappa, and reevaluation of kappa for small to moderate earthquakes at seismic stations in the vicinity of Yucca Mountain, Nevada

We address two primary questions. First, what are the values of seismic spectral decay parameter kappa at stations on and around Yucca Mountain. Second, can small and/or moderate earthquakes provide estimates of kappa that are valid for conditions of strong ground motion