Navegando por Palavras-chave "Western blot analysis"

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    Assessment of gliadin in supposedly gluten-free foods prepared and purchased by celiac patients
    (Lippincott Williams & Wilkins, 2001-01-01) Sdepanian, Vera Lucia [UNIFESP]; Scaletsky, Isabel Cristina Affonso; Fagundes Neto, Ulysses [UNIFESP]; Morais, Mauro Batista de [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Background: the present study was designed to evaluate the presence of gliadin in homemade foods prepared by patients with celiac disease and/or their relatives, as well as in processed products consumed by such patients in São Paulo, Brazil, by enzyme immunoassay (EIA) and Western blot (WB) analysis.Methods: One hundred ninety samples were analyzed: 108 homemade foods prepared in homes of patients with celiac disease, 81 processed products, and 1 positive control of homemade food. All samples were analyzed by EIA based on monoclonal antibodies to heat stable omega -gliadins and related prolamins from wheat. rye, and barley. Samples were also analyzed using the WE technique.Results: Only one (0.9%) of 108 homemade foods contained detectable amounts of gliadin, as determined by EIA. Twelve of 81 processed products contained gliadin by EIA, as follows: 5 of 61 without gluten listed in the ingredients, 2 of 11 malt extracts, 1 of 2 wheat starches, 1 of 2 types of beer. and all 3 positive control products. Gliadin content of these products was between 4 and 10 mg of gliadin/100 g of product, except for the wheat starch sample (28 mg of gliadin/100 g) and all 3 samples with gluten (>4000 mg of gliadin/100 g). the positive control of homemade food contained 152 mg of gliadin/100 g. One hundred three of 190 samples were analyzed by WE, and 21 of these were gliadin positive. A comparison of results obtained by EIA and WE showed no statistical differences between the methods.Conclusions: the greater part of the foods prepared in homes of patients with celiac disease and most processed products supposed to be gluten-frer did not contain gliadin. Therefore, celiac patients adequately prepare gluten-free homemade food and have the expertise to purchase processed gluten-free food in São Paulo, Brazil.
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    A Research Design for the Quantification of the Neuropeptides Substance P and Calcitonin Gene-Related Peptide in Rat Skin Using Western Blot Analysis
    (Lippincott Williams & Wilkins, 2015-06-01) Lapin, Guilherme Abbud Franco [UNIFESP]; Hochman, Bernardo [UNIFESP]; Nishioka, Michele Akemi [UNIFESP]; Maximino, Jessica Ruivo [UNIFESP]; Chadi, Gerson [UNIFESP]; Ferreira, Lydia Masako [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); IB CNPq Brazilian Natl Council Sci & Technol Dev
    OBJECTIVE: To describe and standardize a protocol that overcomes the technical limitations of Western blot (WB) analysis in the quantification of the neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP) following nociceptive stimuli in rat skin.DESIGN: Male Wistar rats (Rattus norvegicus albinus) weighing 250 to 350 g were used in this study. Elements of WB analysis were adapted by using specific manipulation of samples, repeated cycles of freezing and thawing, more thorough maceration, and a more potent homogenizer; increasing lytic reagents; promoting greater inhibition of protease activity; and using polyvinylidene fluoride membranes as transfer means for skin-specific protein. Other changes were also made to adapt the WB analysis to a rat model.SETTING: University research center.MAIN OUTCOME MEASURE: Western blot analysis adapted to a rat model.RESULTS: This research design has proven effective in collecting and preparing skin samples to quantify SP and CGRP using WB analysis in rat skin.CONCLUSION: This study described a research design that uses WB analysis as a reproducible, technically accessible, and cost-effective method for the quantification of SP and CGRP in rat skin that overcomes technical biases.