375 research outputs found

    Production of an egg yolk antibody against Parietaria judaica 2 allergen

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    Abstract Specific antibodies are essential tools for studying proteins as well as for diagnostic research in biomedicine. The egg yolk of immunized chicken is an inexpensive source of high-quality polyclonal antibodies. The 12-kDa Parietaria judaica 2 allergen was expressed as a fusion protein and was used to immunize Leghorn chickens. In this paper, we show, using 2-dimensional gel electrophoresis and immunoblotting, that chicken antibodies raised against a recombinant allergen can be used to recognize similar proteins from a pollen raw extract. Allergen identity was confirmed by nanoLC-nanospray-tandem mass spectrometry analysis. Our data demonstrate for the first time that a synergistic combination of molecular biology, 2-dimensional PAGE, and use of nonmammalian antibodies represents a powerful tool for reliable identification of allergens

    Contribution of proteomics to understanding the role of tumor-derived exosomes in cancer progression: State of the art and new perspectives

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    Exosomes are nanometer-sized vesicles (40–100 nm diameter) of endocytic origin released from different cell types under both normal and pathological conditions. They function as cell free messengers, playing a relevant role in the cell–cell communication that is strongly related to the nature of the molecules (proteins, mRNAs, miRNAs, and lipids) that they transport. Tumor cells actively shed exosomes into their surrounding microenvironment and growing evidence indicates that these vesicles have pleiotropic functions in the regulation of tumor progression, promoting immune escape, tumor invasion, neovascularization, and metastasis. During the last few years remarkable efforts have been made to obtain an accurate definition of the protein content of tumor-derived exosomes (TDEs) by applying MS-based proteomic technologies. To date, TDEs proteomic studies have been mainly utilized to catalog TDEs proteins with the purpose of identifying disease biomarkers. The future challenge for improving our understanding and characterization of TDEs will be the implementation of new systemsdriven and proteomic integrative strategies. The aim of this article is to provide an overview of the most characterized exosomes-mediated mechanisms that contribute to the pathogenesis of cancer and to review recent proteomics data that support the protumorigenic role of TDEs

    Application of an immunoproteomic approach to detect anti-profilin antibodies in sera of paritaria judaica allergic patients

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    Pollen from grasses, weeds, and trees constitutes one of the main sources of inhalant allergens frequently associated with seasonal patterns of allergic diseases. Pollen allergens show some analogies in the amino acids sequence which determine immunological similarity and cross reactivity. Parietaria judaica (P.j) pollen represents one of the main sources of allergens in the Mediterranean area and its major allergens have already been identified (Par j 1 and Par j 2). Recently, has been also described a minor allergen, profilin (Par j 3), an allergen present in pollen of trees, grasses and weeds. Allergenic plant profilins constitute a highly conserved family with sequence identities of 70% to 85% responsible for a wide range of cross-reactivity among pollens and plant foods. In this work we use an immunoproteomic approach to detect IgE antibodies against profilin in serum of P.j allergic patients

    Sedimentology and composition of sands injected during the seismic crisis of May 2012 (Emilia, Italy): clues for source layer identification and liquefaction regime

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    InMay 2012widespread sand blows formed along buried channels in the eastern sector of the Po Plain (Northern Italy) as a consequence of a series of seismic eventswith main shocks ofMw6.1 and 5.9. At San Carlo (Ferrara) a trench dug a few week after the earthquakes exposed sand dikes cutting through an old Reno River channel– levee system that was diverted in the 18th century and was deposited starting from the 14th century (unit A). This sequence overlies a Holocene muddy floodplain deposits and contains scattered sandy channel deposits (unit B) and a Pleistocene channel sand unit (unit C). Sands with inverse and normal grading, concave layering and vertical lamination coexisting along the dikes suggest multiple rhythmic opening and closing of the fractures that were injected and filled by a slurry of sand during the compression pulses, and emptied during the extension phase. The pulse mechanism may have lasted for several minutes and formed well stratified sand volcanoes structures that formed at the top of the fractures. Sands fromdikes and fromthe various units showwell defined compositional fields from lithoarenitic to quartz-feldspar-rich compositions. Sands from the old Reno levee and channel fill (unit A) have abundant lithic fragments derived fromthe erosion of Apennine sedimentary carbonate and terrigenous successions. Composition of the sand filling the dikes showclear affinities with sand layers of the old Reno River channel (Unit A) and clearly differ from any sand from deeper Holocene and Pleistocene layers (Unit B and C),which are richer in quartz and feldspar and poorer in sedimentary lithic fragments. Sorting related to sediment flux variations did not apparently affect the sand composition across the sedimentary structures. Textural and compositional data indicate that the liquefaction processes originated from a relatively shallow source consisting of channel sands located within Unit A at 6.8.to 7.5 m depth

    Proteomic analysis of Parietaria judaica pollen and allergen profiling by an immunoproteomic approach

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    Parietaria judaica pollen is a common cause of airway allergic disease in the Mediterranean area. Proteome analysis of mature Parietaria judaica pollen by two-dimensional gel electrophoresis (2-DE) and mass spectrometry has established the first reference proteome map of this weed. Proteins involved in a variety of cellular functions as well as the occurrence of allergens were detected. By using 2-DE and immunoblotting with sera from Parietaria judaica allergic patients we obtained a more detailed characterization of Parietaria judaica allergen profil

    I distretti del Mezzogiorno

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    CAP. 1. Definizione e caratteristiche del distretto; CAP. 2. Come sono nati i distretti nel Mezzogiorno: fattori di successo ed insuccesso;2.1.Lo sviluppo dei distretti del Mezzogiorno: Brevi cenni storici; CAP. 3. I Distretti del Mezzogiorno; Il "Triangolo del Salotto"; Il polo calzaturiero aversano; La corsetteria di Lavello ;Il TAC del tacco Il distretto barlettano delle calzature; L'abbigliamento della Puglia centrale; La percezione del fenomeno "distretto" dei piccoli e medi imprenditori in Campania, il caso di S. Giuseppe vesuviano e Solofra; Il distretto industriale di Solofra; Conclusioni

    Three-Dimensional Cell Cultures: The Bridge between In Vitro and In Vivo Models

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    Although historically, the traditional bidimensional in vitro cell system has been widely used in research, providing much fundamental information regarding cellular functions and signaling pathways as well as nuclear activities, the simplicity of this system does not fully reflect the heterogeneity and complexity of the in vivo systems. From this arises the need to use animals for experimental research and in vivo testing. Nevertheless, animal use in experimentation presents various aspects of complexity, such as ethical issues, which led Russell and Burch in 1959 to formulate the 3R (Replacement, Reduction, and Refinement) principle, underlying the urgent need to introduce non-animal-based methods in research. Considering this, three-dimensional (3D) models emerged in the scientific community as a bridge between in vitro and in vivo models, allowing for the achievement of cell differentiation and complexity while avoiding the use of animals in experimental research. The purpose of this review is to provide a general overview of the most common methods to establish 3D cell culture and to discuss their promising applications. Three-dimensional cell cultures have been employed as models to study both organ physiology and diseases; moreover, they represent a valuable tool for studying many aspects of cancer. Finally, the possibility of using 3D models for drug screening and regenerative medicine paves the way for the development of new therapeutic opportunities for many diseases
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