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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science, Ltd
    Clinical & experimental allergy 31 (2001), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The main allergens from privet and lilac pollens, Lig v 1 and Syr v 1, are proteins homologous to Ole e 1 and have been shown to be involved in cross-reactivity.To overproduce the correctly folded Lig v 1 and Syr v 1 allergens and to study their immunological properties in comparison with those of their natural counterparts.The yeast Pichia pastoris was used as an expression system to produce these recombinant allergens. The proteins were isolated by ion-exchange and size-exclusion chromatographies. Amino acid quantifying, Edman degradation, mass spectrometry and circular dichroism were carried out to obtain molecular properties of the recombinant proteins. Anti-Ole e 1 monoclonal and polyclonal antibodies, as well as sera from patients allergic to olive pollen, were used in immunoblotting and ELISA for immunological characterization.Recombinant Lig v 1 and Syr v 1 were secreted at high yield to the extracellular medium of the yeast. The purified proteins displayed the native conformation, as deduced from their spectroscopic properties and binding ability to an IgG monoclonal antibody. The recombinant allergens behaved similarly to their natural counterparts when they were analysed against Ole e 1-specific antibodies. IgE and IgG binding properties of lilac and privet allergens to olive allergic sera and Ole e 1-specific antibodies indicated that these molecules share common B-cell epitopes with Ole e 1. P. pastoris yeast is an appropriate system for the efficient production of Ole e 1-like allergens, which could be used as analogous allergens and predictors of clinical sensitization.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Copenhagen : Munksgaard International Publishers
    Allergy 55 (2000), S. 0 
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background: Sera of patients allergic to olive (Olea europaea) pollen were used to analyze the IgE cross-reactivity between olive-pollen extract and other pollens obtained from phylogenetically unrelated species. Methods: We used IgE immunostaining of pollen extracts blotted to nitrocellulose membranes after SDS–PAGE and inhibition analysis of this binding. Results: A high inhibition of the IgE binding on olive-pollen extract was exhibited by birch, mugwort, pine, and cypress pollens, suggesting that these extracts contain proteins which share common epitopes and thus can be recognized by olive-allergic sera. IgE binding to Gramineae pollen extracts was not inhibited by olive-pollen extract, indicating a primary sensitization of the patients to these species. From the inhibition assays, the presence of an allergen of 45 kDa in the olive pollen, which has no homologous counterparts in other allergenic species, has been inferred. Conclusions: Olive pollen contains allergens which cross-react with pollens from unrelated species, a fact that could simplify the diagnosis and treatment of pollinosis.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 178 (2000), S. 175-183 
    ISSN: 1432-1424
    Keywords: Key words: Aging — Skeletal muscle — Single fiber —Soleus—Extensor digitorum longus—Flexor digitorum brevis— Excitation-contraction coupling — Sarcopenia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. In the present study we measured, for the first time, the isometric specific force (SF, force normalized to cross sectional area) generated by single intact fibers from fast- (extensor digitorum longus, EDL) and slow-twitch (soleus) muscles from young adult (2–6), middle-aged (12–14) and old (20–24 month-old) mice. SF has also been measured in single intact flexor digitorum brevis fibers from young mice. Muscle fibers have been classified into fast- or slow-twitch based on the contraction kinetics. Maximum SF recorded in EDL and soleus fibers from young and middle-aged mice did not differ significantly. A significant age-dependent decline in maximum SF was recorded in EDL and soleus fibers from young or middle-aged to old mice. The SF was 377 ± 18, 417 ± 20 and 279 ± 18 kPa for EDL fibers from young, middle-aged and old mice, respectively and 397 ± 17, 405 ± 24 and 320 ± 33 kPa for soleus fibers from age-matched mice, respectively. The frequency needed to elicit maximum force in EDL and soleus fibers from middle-aged to old mice did not differ significantly. In conclusion, the specific force developed by both fast and slow-twitch single intact muscle fibers declines with aging and more significantly in the former.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1615-5947
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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