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  • 1
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Cross-reactivity between the different components in Parietaria judaica pollen extract has been investigated by polyclonal as well as monoclonal antibodies before and after chemical deglycosylation obtained by trifluoromethanesulphonic acid (TFMS) treatment of the extract. In western blotting a polyclonal rabbit antiseruni, obtained by injecting purified Par j I, was able to recognise many components of the native extract. However, its reactivity was restricted, after chemical deglycosylation of the extract, to the major allergen alone, indicating that its cross-reactivity was due to sugar moieties. Moreover, out of several monoclonal antibodies raised by injecting the whole Parietaria judaica extract, one (1A4/2F8) was also able in western blotting to recognise an epitope shared by many components of the extract except the major allergen Par j I. However, in this case the broad reactivity of the antibody was not affected by the deglycosylating procedure. When the reactivity of Parietaria Judaica extract was tested before and after sugar removal, against specific IgE from a pool of patient sera, no differences could be demonstrated, thus indicating that carbohydrates are not strongly involved in the binding of Parietaria judaica-specific IgE. The results indicate that both proteic and carbohydratic cross-reactive epitopes are shared by many components of Parietaria judaica pollen extract.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Olea europaea (olive) pollen extract was prepared by aqueous extraction and characterized by biochemical and immunochemical methods. Two components, displaying respective mol. wt. of 17000 and 19000, were the most reactive allergens, being the doublet (designated Ole e I) recognized by most sera tested. The 19000 mol. wt. component, purified by conventional biochemical procedure and lectin-affinity chromatography from the Ole e I doublet, was deglycosylated and analyzed by SDS–PAGE and by ELISA inhibition. The results obtained suggest that the 19000 mol. wt. component represents the glycosylated form of the 17000 component.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2048
    Keywords: Cicer ; De-etiolation ; Diamine oxidase ; Hydrogen peroxide production (cell wall) ; Peroxidase ; Wound healing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The activities of diamine oxidase (DAO, EC 1.4.3.6) and peroxidase (POD, EC 1.11.1.7) were determined along the stems of light-grown Cicer arietinum L. (chick-pea) seedlings. Enzyme activities were evaluated in the soluble, lightly bound (salt extraction) and tightly bound (Driselase digestion) wall fractions, and in residual fractions obtained from the different internodes. Apparent tissue distributions of both enzymes and lignin depositions were visualised by means of histochemical and immunohistochemical techniques. A close relationship was found between DAO and POD activities in the soluble and wall fractions along the stem. The biochemical activities of both enzymes decreased from the base to the apex of the stem in parallel with the distribution pattern of lignifying tissues in this organ. A similar activity gradient was found for each enzyme along the epidermis of the whole organ. Moreover, deetiolation elicited a rise in the activities of both enzymes in this tissue. Wounding chick-pea stems induced parallel increases in DAO and POD activities in the soluble and wall fractions. In-situ histochemical detection of both enzymes demonstrated the parallel occurrence of the DAO/POD system and lignosuberised depositions in the cell walls adjacent to the wound site. The patterns of POD isoforms resulting from the wound-healing process were determined by means of starch-gel electrophoresis. In addition to changes in relative intensity of enzyme bands in soluble and wall fractions, a new POD isoform, possibly related to the wounding response, appeared in the soluble fraction. This isoform was shown to be lightly bound to cell walls as it could be detected in the extracellular fluids obtained from wound-healed seedlings. On the basis of the above-mentioned results, a strict spatial and functional correlation can be inferred between DAO and POD in chick-pea, and probably in other Leguminosae species, in accordance with previous evidence indicating an integrated role for these enzymes in the apoplast.
    Type of Medium: Electronic Resource
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