Poster
# 34
Poster |
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6th Internet World Congress for Biomedical Sciences |
Maria Rosa Sánchez(1), Manuel Samaniego(2)
(1)Departamento de Bioquímica.. Hospital Universitario "San Cecilio". Granada - Granada. Spain
(2)Unidad Neonatal. Hospital Universitario "Virgen de las Nieves". - Granada. Spain
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[Biochemistry] |
Alkaline phosphatase (ALP) is a membrane-bound enzyme that is found in association with virtually all cells. ALP comprising four isoenzymes encoded by different structural genes: tissue nonspecific (bone, liver, renal), intestinal, placental, and germ cell (testis, thymus and lung). ALP in the lung has been localized to the plasma membrane and lamellar bodies of type II alveolar epithelial cells and its concentration in lung fluids has been used as a marker of damage of these cells.
Purpose of study: The aim of this study was to test the utility of alkaline phosphatase isoenzyme determinations, as injury markers of cellular membranes from bronchial ephiteliun, in patients diagnosed with bronchopulmonary pathology from different etiology, using serum and bronchoalveolar lavage fluid (BALF) samples.
Study population: 62 normal subjects and 80 patients affected by different pulmonary diseases were grouped according to age as following:
1) 26 preterm neonates suffering Respiratory Distress Syndrome (RDS) requiring mechanical ventilation and 20 healthy newborns of similar gestational ages and birth weights.
2) 44 children, rangin from 2 to 12 years old: 12 healthy and 32 with respiratory problems (18 obstructive bronchitis from alergical causes, and 15 inflamatory process by infection agents).
3) 52 adults (30-65 years old): 30 control subjets and 22 patients with bronchopulmonary pathology and infiltrates of doubtful cause in chest radiograph, all of them examined by bronchoscopy for the purpose of diagnosis.
enzyme assays: The bronchial aspirate was collected by standard methods after bronchial lavage with saline solution at 0,9%, and centrifuged to eliminate mucus. The supernatant was used for the analysis. The total activity of ALP was assayed at 37ºC by a multiparametric analyzer Hitachi 917. The isoenzymes were separated by electrophoresis on agarose gel with Paragon Beckman System Isopal, using confirmatory methods (thermodenaturation, incubation with neuraminidase, ficin and urea, treatment with antiplacental antiserum, precipitation with lecitin of wheat germen), quantified by densitometry and their activity expressed as a percentage of the total ALP. Total proteins were quantified in the BALF and ALP enzymatic activity expressed in U/mg of protein × 10-³
We found that macromolecular ALP fraction (HMW-ALP) was significantly increased in the serum of neonates with distress (p<0.01), in the children (p<0.001), and adults affected by pulmonary pathology (p<0.001).
The electrophoresis of ALP isoenzymes showed an unique isoenzymatic band corresponding with its macromolecular fraction in the bronchoalveolar lavage fluid.
We conclude that increased macromolecular fraction of alkaline phosphatase found in the serum of patients diagnosed with respiratory problems could have its origin in damaged pulmonary tissue.
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[Biochemistry] |
