Expression of the apoptosis inhibitor Bcl-2 in sputum eosinophils from children with acute asthma.

BACKGROUND: Apoptosis of eosinophils is of increasingly important value in modulating allergic airway inflammation in asthma. Our purpose was to investigate the degree of expression of the antiapoptotic B-cell lymphoma/leukaemia-2 (Bcl-2) protein in sputum eosinophils during acute asthma exacerbation and its relationship with exacerbation severity. METHODS: Sputum was obtained from 33 asthmatic children and 15 healthy children as a control group. Patients were studied during an acute asthma exacerbation. They were classified according to the severity of exacerbation into mild, moderate and severe (n=11 for each). Patients with severe exacerbation were followed up until remission and another sputum sample was obtained. Number of sputum eosinophils was expressed as percentage of leucocytes. Bcl-2 expression in sputum eosinophils was assessed by immunohistochemical staining techniques; the results were expressed as percentage of positively stained cells over total eosinophils. RESULTS: Sputum eosinophils and Bcl-2(+) eosinophils' percentages were significantly higher in patients with acute exacerbation than controls (P<0.01). Patients with severe exacerbation had significantly higher sputum Bcl-2(+) eosinophils' percentage than those with mild-to-moderate exacerbation (mean+/-SD=42.4+/-31.96% vs. 5.7+/-14.5%, P<0.01). A significant negative correlation was found between Bcl-2(+) eosinophils' percentage and peak expiratory flow rate % predicted (P<0.05). After remission, patients with severe exacerbation showed a significant decrease of Bcl-2(+) eosinophils' percentage (P<0.05). CONCLUSION: Our findings suggest that Bcl-2 prolongs survival and decreases apoptosis of airway eosinophils in asthma especially during exacerbation. Eosinophil apoptosis and Bcl-2 represent a target for new and effective therapeutic strategies of asthma.

Identification of novel Asian Indian and Japanese mutations causing beta-thalassaemia in the Egyptian population.

beta-thalassaemia is a major health problem in Egypt. It has been estimated that of the 1.5 million live births. 1000 children with beta-thalassaemia major are born annually. Although the available treatment has increased the life expectancy of patients, it is still unsatisfactory and represents a significant drain on the country's resources. National screening and prenatal diagnosis programmes can be provided in Egypt once the spectrum of beta-thalassaemia mutations has been identified within the Egyptian population. We have examined 16 DNA samples with 21 beta-thalassaemia mutations that remained unidentified in a study of 54 patients reported by Rady and colleagues in 1996. Using the polymerase chain reaction and single strand conformation analysis we identified the following changes: frameshift (FS) codon (CD) 8/9 (+G), 4 FS CD 29 (-G) and 2 novel mutations in exon I (15 CD 22 A-C and 1 FS CD 28 -C). In addition, a silent, probably polymorphic mutation, CD 17 G-A was present in all chromosomes.

Identification of Mediterranean beta-thalassemia mutations by reverse dot-blot in Italians and Egyptians.

beta-Thalassemia is a significant public health problem in Egypt where over 1000 of the annual 1.5 million newborns are expected to be affected with this disorder. A preventive program of the disease should be multifaceted with its technical component based on carrier screening and prenatal diagnosis through mutation detection. In addition, it should have an information and educational component with the aim of increasing public awareness of the disease. Proper selection of the technique(s) to be utilized in such a program is highly important. The appropriate technique to be used in screening should be reliable, simple and cost effective. It should also circumvent the problem of marked heterogeneity of the disease in Egypt. The reverse dot-blot technique has been used in the present study for the characterization of mutations in 138 Italian and 108 Egyptian thalassemia chromosomes, confirming its reliability as a screening method. The technique is now in routine use for thalassemia diagnosis in the Microcitemia Center of the Galliera Hospital in Genoa, Italy. Based on these results, we recommend the reverse dot-blot method as the technique of choice in the preventive program of this disease in Egypt.