One- and two-dimensional electrophoretic identification of IgE-binding polypeptides of Lupinus albus and other legume seeds

The prevalence of food allergies in the world population requires integrated approaches to identify new potential allergens, especially those of plant origin. The aim of this work was the allergen in vitro analysis of Lupinus albus seed proteome, a promising food protein source, and the assessment of IgE cross-reactivities with other more diffused legume species. A combination of one- and two-dimensional gel electrophoresis and immunoblotting analyses with specific IgGs for band identification and lupin-sensitized patients' circulating IgEs for allergenicity studies has been used. Two lupin proteins, namely, conglutin gamma and 11S globulin basic subunits, strongly reacted with all patients' sera. Also, cross-reactivities with the homologous polypeptides of other legume species were observed. Otherwise, no reaction at all was detected with a 2S-type lupin protein. This global electrophoretic approach has allowed the identification of a new potential lupin allergen and confirmed the cross-reactivity among the legume 11S globulin basic subunits.     

Simultaneous determination of Cd(II), Cu(II), Pb(II), and Zn(II) in citrus essential oils by derivative potentiometric stripping analysis

Citrus essential oils are widely used in the food, cosmetics, and pharmaceutical industries, so the determination of heavy metals content is of great importance to guarantee their quality. The present work deals with the quantification of Cd(II), Cu(II), Pb(II), and Zn(II) in different varieties of citrus essential oils, using derivative potentiometric stripping analysis. Two different metals extraction procedures, involving concentrated hydrochloric acid treatment and acid-alcoholic dissolution, are tested on lemon, mandarin, sweet orange, and bergamot essential oils, and they give very similar results. Cd(II), Cu(II), Pb(II), and Zn(II) recovery tests spanned from 95 to 100.50%, providing evidence that metals quantification remained unaffected by the cleanup steps of the two procedures. The repeatability of the hydrochloric acid extraction method, applied on different varieties of essential oils, is >95.00% for Cd(II), Cu(II), Pb(II), and Zn(II), whereas the repeatability of the acid-alcoholic dissolution method is >93.00% for Cu and Cd only in lemon oil. Detection limits obtained for the four analytes, using both procedures, ranged from 0.10 to 0.98 ng g(-)(1) in lemon, mandarin, sweet orange, and bergamot essential oils.     

A New Bioassay Platform Design for the Discovery of Small Molecules with Anticancer Immunotherapeutic Activity

Immunotherapy takes advantage of the immune system to prevent, control, and eliminate neoplastic cells. The research in the field has already led to major breakthroughs to treat cancer. In this work, we describe a platform that integrates in vitro bioassays to test the immune response and direct antitumor effects for the preclinical discovery of anticancer candidates. The platform relies on the use of dendritic cells that are professional antigen-presenting cells (APC) able to activate T cells and trigger a primary adaptive immune response. The experimental procedure is based on two phenotypic assays for the selection of chemical leads by both a panel of nine tumor cell lines and growth factor-dependent immature mouse dendritic cells (D1). The positive hits are then validated by a secondary test on human monocyte-derived dendritic cells (MoDCs). The aim of this approach is the selection of potential immunotherapeutic small molecules from natural extracts or chemical libraries.     

Science mapping approach to analyze the research evolution on precision agriculture: world,EU and Italian situation

The relevance of precision agriculture produced a growth of the related literature over the years. However, a structured analysis of the published material is still missing. Thus, this study attempts to analyze the global scientific output of precision agriculture researches published during the period 2000–2016. By using a science mapping approach, mainly based on the application of network analysis tools, it was possible to investigate pivotal aspects of this research field such as publication trends, research topics and their geographical distribution. Using the Scopus database 17,756 scientific publications were retrieved from the chosen period. The number of publications increased after 2006, highlighting the vibrancy of the field. By authoring 35% of the publications, U.S.A. and China were the most active knowledge producer countries. Moreover, the generation of time resolved maps allowed us to identify agriculture engineering, computer science and agriculture studies as three main research areas characterizing precision agriculture panorama. The paper discusses the distribution of these topics at global level, among European countries and in Italy. Overall, this analysis represents the first holistic view of precision agriculture research providing valuable information for farmers, policy makers and researchers.     

Emergence and early evolution of fungicide resistance in North American populations of Zymoseptoria tritici

Although fungicide resistance in crop pathogens is a global threat to food production, surprisingly little is known about the evolutionary processes associated with the emergence and spread of fungicide resistance. Early stages in the evolution of fungicide resistance were evaluated using the wheat pathogen Zymoseptoria tritici, taking advantage of an isolate collection spanning 20 years in Oregon, USA, and including two sites with differing intensity of fungicide use. Sequences of the mitochondrial cytb protein conferring single‐mutation resistance to QoI fungicides and the nuclear CYP51 gene implicated in multiple‐mutation resistance to azole fungicides were analysed. Mutations associated with resistance to both fungicides were absent in the 1992 isolates, but frequent in the 2012 collection, with higher frequencies of resistance alleles found at the field site with more intensive fungicide use. Results suggest that the QoI resistance evolved independently in several lineages, and resulted in significant mitochondrial genome bottlenecks. In contrast, the CYP51 gene showed signatures of diversifying selection and intragenic recombination among three phylogenetic clades. The findings support a recent emergence of resistance to the two fungicide classes in Oregon, facilitated by selection for mutations in the associated resistance genes.     

Protective effect of the AT137RQ and ARQK176 PrP alleles against classical scrapie in Sarda breed sheep

The susceptibility of sheep to scrapie is under the control of the host’s prion protein (PrP) gene and is also influenced by the strain of the agent. PrP polymorphisms at codons 136 (A/V), 154 (R/H) and 171 (Q/R/H) are the main determinants of susceptibility/resistance of sheep to classical scrapie. They are combined in four main variants of the wild-type ARQ allele: VRQ, AHQ, ARH and ARR. Breeding programmes have been undertaken on this basis in the European Union and the USA to increase the frequency of the resistant ARR allele in sheep populations. Herein, we report the results of a multi-flock study showing the protective effect of polymorphisms other than those at codons 136, 154 and 171 in Sarda breed sheep. All ARQ/ARQ affected sheep (n = 154) and 378 negative ARQ/ARQ controls from four scrapie outbreaks were submitted to sequencing of the PrP gene. The distribution of variations other than those at the standard three codons, between scrapie cases and negative controls, was statistically different in all flocks. In particular, the AT₁₃₇RQ and ARQK₁₇₆ alleles showed a clear protective effect. This is the first study demonstrating a protective influence of alleles other than ARR under field conditions. If further investigations in other sheep breeds and with other scrapie sources confirm these findings, the availability of various protective alleles in breeding programmes of sheep for scrapie resistance could be useful in breeds with a low frequency of the ARR allele and would allow maintaining a wider variability of the PrP gene.     

Grain legume proteins and nutraceutical properties

Grain legumes are a valuable source of food proteins. Their exploitation is expected to grow in relation of a growing world's food needs. Moreover, it is currently taking place a reappraisal of the beneficial effects of legume seed dietary intake, which are the basis for various health claims. Proteins and peptides concur to the observed biological activities of legume seeds, but their effect(s) has(ve) not completely been disclosed. Aims of this review are: to report the most relevant putative positive effects of grain legumes on human health and to give an account of the current knowledge on the demonstrated legume seed protein biological activities. Specific effects on the prevention and treatment of various diseases, mostly of which are typical of the affluent countries, are reported. Examples of studies at molecular level aimed at elucidating of the underlying mechanism(s) are given. The prospects on targeted legume protein exploitation in the nutraceutical area, including the biotechnological approaches, are also considered.     

IgE-Mediated Cross-Reactivity among Leguminous Seed Proteins in Peanut Allergic Children

The immunological cross-reactivity among major protein- and oil-crops, including lupin, lentil, pea, peanut, kidney bean and soybean, has been studied by a combination of in vitro and in vivo experimental approaches: SDS-PAGE separations of legume protein extracts and immuno-blot revelations with 12 peanut-sensitive subjects’ sera, Immuno-CAP and Skin Prick tests on the same subjects. The immuno-blotting data showed a wide range of IgE-binding responses both displayed by one subject towards different plant extracts and among subjects. Differences were both quantitative and qualitative. The prevalent responses of most subjects’ sera were seen with peanut polypeptides, as expected, as well as with various polypeptides of the other legumes, the most recurrent of which were the basic subunits of the 11S globulins. The distribution of in vivo responses generally paralleled those obtained by in vitro approaches with strong responses elicited by peanut, lentil and pea protein extracts, especially by most sensitive subjects, thus providing a consistent overall set of results. In this work, the comparison of various approaches has allowed us to get an overall broad picture of the immunological cross-reactivities among proteins of widely used different seed species and to hypothesize the role of most conserved specific polypeptides.     

Effects of corn deficit irrigation and soil properties on water use efficiency. A 25-year analysis of a Mediterranean environment using the STICS model

This study analyses the role of variability induced by climate and soil properties (texture and total available soil water in the root zone, TAW) on the “corn deficit irrigation–water use efficiency” relation over a 25-year period, between 1981 and 2005, in three different sites located in the South of Italy. The analysis of water use efficiency was carried out by means of three indexes: crops water use efficiency (WUE_ET and WUE_T) and irrigation water use efficiency (IWUE). These indexes can be obtained by calculating the ratio between the yield and the seasonal values of evapotranspiration (ET), transpiration (T) and irrigation volume (I). After its validation, the STICS model was retained to simulate the variables required to determine these indexes.Climatic variability affected the three indexes in different ways during the 25-year period studied. The dispersion around the mean values of the water use efficiency is 17% on average for the WUE (ET or T) indexes, whereas it could reach 54% for the IWUE index. This last index is only poorly reliable due to the high variability of rainfall during the corn growing season in the Mediterranean region.For the same level of soil water deficit, TAW led to an improvement (the case of the site with a high TAW and loam–clay soil texture), a stabilisation (the case of the site with a high TAW and clay soil texture) or a decrease (the case of the site with a poor TAW and clay soil texture) in WUE_ET and WUE_T values. For the same conditions of soil water deficit and TAW, crops water use efficiency was higher in loam textured soil than in clay soil.The results derived by analyzing three indexes (WUE_ET, WUE_T and IWUE) converge to the same conclusions on the “corn deficit irrigation–water use efficiency” relation observed at the three different sites.The hypotheses which may explain the differences observed in the different sites studied in terms of water use efficiency are discussed, with the presentation of some suggestions devised for corn irrigation practices.