Print version ISSN 0034-7299
Rev. Bras. Otorrinolaringol. vol.75 no.2 São Paulo Mar./Apr. 2009
Gesmar Rodrigues Silva SegundoI; Mônica Camargo SopeleteII; Sílvia Azevedo TerraIII; Fernando Lourenço PereiraIV; Caroline Morais JustinoV; Deise Aparecida de Oliveira SilvaVI; Ernesto Akio TaketomiVII
IMaster's degree, doctoral student in immunology and applied parasitology, Universidade Federal de Uberlandia
IIDoctorate, post-doctoral student in allergy and clinical immunology, Universidade Federal de Uberlandia
IIIMaster's degree, assistant professor of veterinary immunology, Ituverava, SP
IVMaster's degree, doctoral student in applied immunology, FMRP/USP
VMaster's degree, basic education teacher, Santos, SP
VIDoctorate, researcher and supervisor of the Allergy and Clinical Immunology Laboratory, Universidade Federal de Uberlandia
VIIDoctorate / post-doctorate, full professor of immunology and head of the Allergy and Clinical Immunology Laboratory, Universidade Federal de Uberlandia. Universidade Federal de Uberlandia
The prevalence of allergic diseases such as asthma, rhinitis, allergic conjunctivitis and atopic dermatitis has increased in the last decades. The relationship between allergen exposure, atopic sensitization and development of allergic diseases is widely described in the literature.
AIM: To evaluate measures for reducing allergen exposure as part of the treatment of allergic diseases.
METHODS: An analysis was made of previous studies on allergen exposure done with a similar methodology in the central region of Brazil; the study included homes, hotels, cinemas, cars, taxis, buses and scholar transportation.
Results: High levels of Der p 1 and Der f 1 mite allergens were found in a large proportion of the sample in most of the environments included in those studies; there were higher levels of pet allergens in cars and school transportation vehicles.
CONCLUSION: The diversity of allergen exposure demonstrates the need for education about allergic diseases for patients and their families, as well as measures of reducing allergens in homes. This should be part of a global strategy of the management of allergic diseases, given that individuals live in society, not only in their houses.
Keywords: aeroallergen, allergen, environment, asthma, allergen exposure, allergic rhinitis.
Atopy is the term for individuals whose immune system produces IgE antibodies in response to specific antigenic stimuli. Allergy is the term for an altered state resulting from an immunological process set in motion by a substance or allergen that is inert1 or that induces a non-IgE mediated antibody response in normal individuals. The majority of allergic diseases is caused by the type I (immediate) hypersensitivity reaction, which is an IgE-mediated abnormal immune response.
The prevalence of allergic diseases such as asthma, rhinitis, allergic conjunctivitis and atopic dermatitis has increased in the past few decades.2 ISAAC data recently described in Brazil show that the prevalence of asthma may reach 28.2%, depending on which center is studied.3 Another Brazilian multicentric study revealed a significant difference in air allergen sensitization between allergic individuals and healthy controls, demonstrating the relation between atopy and these conditions.4 Other authors have shown that there is atopic sensitization in over 40% of young adults in the USA.5
Kern (1921) first described house dust as an allergen; his report showed that many asthma and rhinitis patients developed skin erythema and edema due to dust extracts from their own houses.6 Various sources of house dust allergens had been described by 1960, including animal dander, insects and fungi. In 1967, a mite in dust (Dermatophagoides pteronyssinus) was shown to cause a skin reaction; it has since been considered as the main house dust allergenic agent in the Netherlands, and subsequently in other countries such as the United Kingdom, Australia, Japan and Brazil.7
House dust allergens were purified during the following decades; the cat (Felis domesticus) allergen - Fel d 1 - was the first to be described, in 1974.8 In 1980, the first mite allergen, Der p 1 (derived from Dermatophagoides pteronyssinus), was isolated and characterized.9
Mite detection in house dust was done in the past by microscopy and acarid count per square centimeter; quantification of allergens in different sites, however, was not possible at that time. After the development of monoclonal antibodies (a highly sensitive and specific tool for identifying and quantifying antigens), many authors have standardized immunoenzymatic tests for investigating allergen levels in house dust and environmental extracts, which have underlined environmental prevention measures.10
The relation between exposure, atopic sensitization and the development of allergic disease has been widely described in the literature. Basic evidence for a cause-effect relation between allergen exposure and sensitization arose from studies showing that children in places with no mites in house dust (Northern Sweden, for instance) were not sensitized to mite allergens. Likewise, children brought up with little or no exposure to cockroaches (in places such as New Zealand, Sweden or Delaware-USA) were not sensitized to cockroach allergens.11
Prevalence, allergen exposure, and sensitization studies of asthmatic children and adults have suggested that exposure to over 2µg of Der 1 allergens per gram of dust is a risk factor in genetically predisposed individuals. The allergen risk factor for animal dander (Can f 1, Fel d 1) is exposure to over 1µg/g dust. These values are currently believed to vary significantly, as allergic diseases have been considered as multifactorial conditions that depend on the interaction between many factors, such as inheritance, diet, permanence time in closed environments, age of contact and others.12-16 As allergen contact is one of the factors causing inflammation in allergic diseases (especially in asthma and allergic rhinitis), environment control has become a mainstay in the treatment of these conditions, and has been recommended by national and international consensuses. Various published papers had shown that there are variable responses to allergen control measures; the study design and the response assessments, however, varied among these papers, which has generated controversies among physicians.17-28
The purpose of this paper was to discuss the difficulty of controlling allergen exposure and to review various studies in different environments within which individuals spend their days.
This review was composed of published studies about allergen exposure undertaken in the central region of Brazil. The climate (humidity and temperature) was similar in all studies, as were the allergen dosing techniques in various closed environments. Sampling included 124 houses, 5 movie theater rooms, 20 hotels, 60 private vehicles, 120 interstate passenger buses, 60 taxis and 60 school children transport vans.29-36
After obtaining approval from participants, dust samples were collected in each site by a portable aspirator onto which a paper filter had been adapted to retain the dust. Dust was then placed in a plastic bag and identified; storage was at 4ºC before allergens were extracted. Dust samples were sieved using a standard 0.3 mm pore sieve, Series ASTM (USA), onto a Petri dish and then transferred to test tubes. Allergen fractions were extracted from 100 mg of dust from each sample plus 2 ml of a borate buffered saline solution (BBS), at 5 mM, pH 8.0, at 4ºC, rotatory shaking, during 18 hours. Samples were then centrifuged at 10,000 g for ten minutes; the supernatant was stored at -20ºC for future analysis of its allergen content.
The enzyme-linked immunosorbent assay (ELISA) was used for detecting the Der p 1, Der f 1, Can f 1 and Fel d 1 allergens as described by Luczynska et al.37 modified by Sopelete et al.28 The following monoclonal antibodies (mAb) for capture were used: anti-Der p 1 (clone 5H8), anti-Der f 1 (clone 6A8), anti-Fel d 1 (clone 6F9), and anti-Can f 1 (clone 6E9); the concentration was 10 mg/ml in a carbonate-bicarbonate buffer at 0.06M and pH 9.6. Detection was done using biotinilated mAb, as follows: anti-Der p 1 and anti-Der f 1 (4C1), anti-Fel d 1 (3F4C4) and polyclonal rabbit serum anti-Can f 1 at 1:500. After incubation with streptavidine-peroxidase (Sigma Chemical Co., USA), the test tube was developed by adding an enzymatic substrate (0.01 M ABTS and 0.03% H2O2); reading was done at 405nm. Reference standards containing known levels of each allergen were included into each plate in double to obtain curves for eleven double serial dilutions starting at 250 ng/ml for Der p 1 and Der f 1, 500 ng/ml for Can f 1, and 80ng/ml for Fel d 1. Results were expressed in µg/g of dust.
The Graph Pad Prism version 3.0 (Graph Pad Software, Inc.) software was used for the statistical analysis. The comparison between allergen levels was done using non-parametric tests, as the results did not have a normal (Gaussian) distribution. Geometric means (mg) at a 95% confidence interval (CI) were calculated for each allergen level. the difference between the means was made using the Mann Whitney U test; the significant level was 5% (p < 0.05).
The samples in each environment were analyzed at different times in central inland Brazilian cities (Uberlandia, Uberaba and Goiania); these cities have similar climates (humidity and temperature).
Two studies investigated the presence of household mites in two cities (Uberlandia and Uberaba). The first of these was done in 1998, and verified the presence of sensitizing mite allergen levels in five places within 64 households - sofas, floor of television rooms, beds, floor of bedrooms and kitchens. Elevated levels of Der f 1 were found in the beds of asthmatics (15.8 µg/g dust) and non-asthmatics (8.2 µg/g dust). Der p 1 levels were lower in the beds of these same individuals (asthmatics - 2.8 µg/g dust; non-asthmatics - 4.9 µg/g dust). The mean concentration between five samples showed that 91% of the samples in the homes of asthmatics, and 84% of the samples in the homes of non-asthmatics had Der f 1 levels over 2 µg/g, while 72% of the samples in the homes of asthmatics e 75% of the samples in the homes of non-asthmatics had Der p 1 levels higher than 2µg/g dust.29
In Uberaba, samples for analyzing mite allergens were taken from sofa and bed surfaces (mattresses, bedspreads and pillows) in 60 households in two separate periods (March and July). Higher levels were found in March in these samples; Der f 1 levels were 31.7 µg/g dust in beds and 8.3 µg/g dust on sofas. Der p 1 levels were 0.3 µg/g dust in both months.30
Another study done in Uberlandia assessed the presence of dust allergens in hotels, between June and October 2002; 98 samples from 20 hotels were analyzed. Mean concentrations were: Der f 1 - 11.3 µg/g dust, Der p 1 - 0.15 µg/g dust, Can f 1 - 0.3 µg/g dust and Fel d 1 - 0.11 µg/g dust.31
In Goiania, studies analyzed the surfaces of sofas and floors in five movie theaters. The mean concentrations were: Dep p 1 - 0.35 µg/g dust; Der f 1 - 6.85 µg/g dust; Can f 1 - 0.65 µg/g dust; and Fel d - 10.07 µg/g dust. The allergen Bla g 2 was not detected in any sample.32
Another study investigated interstate transport and taxis; the following mean allergen levels were found in buses with natural ventilation and buses with air conditioning: Der p 1 - 1.6 µg/g dust and 4.3 µg/g dust; Der f 1 - 0.8 µg/g dust and 2.4 µg/g dust; and Fel d 1 - 1.2 µg/g dust and 1.6 µg/g dust. The results in taxis were: Der p 1 - 0.9 µg/g dust; Der f 1 - 1.7 µg/g dust; and Fel d 1 - 1.6 µg/g dust. Although some means were not elevated, the percentage of air conditioned buses with sensitizing allergen levels was 82%, the percentage of naturally ventilated buses with sensitizing allergen levels was 62%, and the percentage of taxis with sensitizing allergen levels was 65%.33
An analysis of samples taken from the seats of passenger vehicles (cars) in Uberlandia found the following mean concentrations: Der p 1 - 0.24 µg/g dust; Der f 1 - 0.29 µg/g dust; Can f 1 - 1.51 µg/g dust; and Fel d 1 - 0.42 µg/g dust.34 The seats of school transportation vans in the same city revealed the following mean concentrations: Der p 1 - 0.15 µg/g dust; Der f 1 - 0.26 µg/g dust; Can f 1 - 1.03 µg/g dust; and Fel d 1 - 0.37 µg/g dust. 35
Table 1 shows the allergen level means according to each site.
The association between allergen exposure, sensitization and allergic diseases has been known since the beginning of the past century. Immunological mechanisms by which allergens cause genetically predisposed individuals to produce allergen-specific IgE and the resulting symptoms - as well as interrelated factors in allergic disease - have been studied in detail in the last two decades. The aim has been to understand the major increased in the prevalence of allergic diseases such as asthma, allergic rhinitis and atopic dermatitis in the past 30 years.
There are two main types of studies for investigating allergenic exposure: (1) those in which individuals move to different, allergen-free environments, and (2) those in which measures to reduce allergen exposure within households are undertaken.
An elegant study in the 1980s, in which asthmatic patients were admitted to hospital for rigorous and prolonged control of allergen exposure - where demonstrably local mite allergen levels were decreased - revealed that bronchial hyperreactivity decreased in these patients.25 Other similar studies assessed changes of environment in which patients were moved to high altitude places where mite levels were low; in these cases, sensitized patients also improved.26-28 A recent prospective study analyzed environment care measures for children at risk since birth, and demonstrated that there was no difference in sensitization to air allergens between the cared group and controls (in which no environmental measures were undertaken); there was, however, a difference in lung function between groups at age 3 years.38
Studies that aimed to decrease allergen exposure in households reached conflicting results. These studies used fundamentally different methods, such as the types of environmental control measures, the sample population, and the form by which results were analyzed.12-20 Most of the studies that showed no difference in the occurrence of symptoms of asthma and allergic rhinitis following environmental hygiene defined environmental control as orientations to reduce household allergens by using mattress covers and other point modifications.
Allergen sensitization depends on individual genetics, the amount of allergens and the duration of exposure. Certain pollens cause allergen sensitization during a relatively short pollination period (less than two week a year). Thus, it may be possible for genetically predisposed individuals - especially in childhood - to become sensitized by a few repetitive exposure periods in sites with elevated mite or animal dander allergen levels.11
The studies that were reviewed in this paper showed that allergens were present in a variety of environments, such as movie theaters, hotels, buses, schoolchildren transport vans, taxis and private vehicles, as well a elevated mite allergen levels in various sites within households. These studies corroborate the association between allergen exposure and sensitization by showing that there were enough allergens to cause sensitization in a variety of commonly used places. These data are similar to those in other studies of allergen exposure that have demonstrated high air allergen levels in nurseries and school, offices and files, hospitals, movie theaters and public transportation vehicles.39-43
Environmental control (tertiary preventive measures) to reduce household air allergens remains a forma recommendation for patients with allergic conditions in which allergen sensitization has been demonstrated. Secondary prevention, which involves environmental control measures done since birth in households of asymptomatic children whose parents have been diagnosed with allergic diseases, is also recommended. These individuals also require vehicle care against allergens. Individuals live in society - not only within their houses - where clinically relevant allergens may be found in places such as schools, workplaces, leisure areas and public transport vehicles. Thus, patients have to not only understand the disease itself, but also the ample possibilities of allergen exposure. Environmental control, therefore, although not sufficient to eliminate the symptoms, should become part of a wider strategy for the treatment of allergic patients and families. It would be wrong to state that environmental control in households by itself does not work in preventing symptoms in allergic patients. Failure of the environmental control measures may occur because it is rare for individuals to be completely free from contact with significant allergen levels, given the diversity of allergen exposure to which every one is subject daily.
1. Johansson SG, Bieber T, Dahl R, Friedmann PS, Lanier BQ, Lockey RF, et al. Revised nomenclature for allergy for global use: Report of the Nomenclature Review Committee of the World Allergy Organization, October 2003. J Allergy Clin Immunol. 2004;113:832-6. [ Links ]
2. Mösges R. The increasing prevalence of allergy: a challenge for the physician. Clin Exp All Rev. 2002;2:13-7. [ Links ]
3. Solé D, Camelo-Nunes IC, Wandalsen GF, Naspitz CK, Vanna AT, Amorim A, et al. A asma em crianças é um problema de saúde pública? Rev Bras Alerg Imunopatol. 2004;27:185-8. [ Links ]
4. Naspitz CK, Sole D, Aguiar MC, Chavarria ML, Rosario Filho N, Zuliani A,et al. Grupo PROAL. Phadiatop in the diagnosis of respiratory allergy in children: Allergy Project-PROAL. J Pediatr. (Rio J) 2004;80:217-22. [ Links ]
5. Burney P, Malmberg E, Chinn S, Jarvis D, Luczynska C, Lai E. The distribution of total and specific serum IgE in the European Community Respiratory Health Survey. J Allergy Clin Immunol 1997;99:314. [ Links ]
6. Kern RA: Dust sensitization in bronchial asthma. Med Clin North Am. 1921;5:751. [ Links ]
7. Platts-Mills TA. Indoor Allergens. Em: N. Franklin Adkinson, Jr., MD, John W. Yunginger, MD, William W. Busse, MD, Bruce S. Bochner, MD, Stephen T. Holgate, MD, DSc, FRCP, FRCPE and F. Estelle R. Simons, MD, PFCPC, editores. Middletons Allergy Principles and Practice. 6th Edition. New York: Mosby; 2003. p. 465-78. [ Links ]
8. Ohman Jr JL, Lowell FC, Bloch KJ. Allergens of mammalian origin. III. Properties of a major feline allergen. J Immunol. 1974;113:1668. [ Links ]
9. Chapman MD, Platts-Mills TA. Purification and characterization of the major allergen from Dermatophagoides pteronyssinus-antigen P1. J Immunol. 1980;125:587. [ Links ]
10. Chapman MD, Sutherland WM, Platts-Mills TA. Recognition of two Dermatophagoides pteronyssinus-specific epitopes on antigen P1 by using monoclonal antibodies: binding to each epitope can be inhibited by serum from dust mite-allergic patients. J Immunol. 1984;133:2488. [ Links ]
11. Platts-Mills TA. Allergen Avoidance. J Allergy Clin Immunol. 2004;113:388-91. [ Links ]
12. Ingram JM, Sporik R, Rose G, Honsinger R, Chapman MD, Platts-Mills TA. Quantitative assessment of exposure to dog (Can f 1) and cat (Fel d 1) allergens: Relation to sensitization and asthma among children living in Los Alamos, New Mexico. J Allergy Clin Immunol. 1995;96:449-56. [ Links ]
13. Gelber LE, Seltzer LH, Bouzoukis JK, Pollart SM, Chapman MD, Platts-Mills TA. Sensitization and exposure to indoor allergens as risk factors for asthma among patients presenting to hospital. Am Rev Respir Dis. 1993;147:573-8. [ Links ]
14. Platts-Mills TAE, Thomas WR, Aalberse RC, Vervloet D, Chapman MD. Dust mite allergens and asthma: report of a second international workshop. J Allergy Clin Immunol. 1992;89:1046-60. [ Links ]
15. Custovic A, Chapman MD. Risk levels for mite allergens. Are they meaningful? Allergy. 1998;53:71-6. [ Links ]
16. Tranter, D.C. Indoor allergens in settled school dust: a review of findings and significant factors. Clin Exp Allergy. 2005;35:126-36. [ Links ]
17. Platts-Mills TAE, Vaughan JW, Carter MC, Woodfolk JA. The role of intervention in established allergy: avoidance of indoor allergens in the treatment of chronic allergic disease. J Allergy Clin Immunol. 2000;106:787-803. [ Links ]
18. Van der Heide S, Kauffman HF, Dubois AEJ, de Monchy JGR. Allergen reduction measures in houses of allergic asthmatic patients: effects of aircleaners and allergen-impermeable mattress covers. Eur Respir J. 1997;10:1217-23. [ Links ]
19. Htut T, Higenbottam TW, Gill GW, Darwin R, Anderson PB, Syed N. Eradication of house dust mite from homes of atopic asthmatic subjects: a double-blind trial. J Allergy Clin Immunol. 2001; 107:55-60. [ Links ]
20. Carswell F, Birmingham K, Oliver J, Crewes A, Weeks J. The respiratory effects of reduction of mite allergen in the bedrooms of asthmatic children-a double-blind controlled trial. Clin Exp Allergy. 1996;26:386-96. [ Links ]
21. Terreehorst I, Hak E, Oosting AJ, et al. Evaluation of impermeable covers for bedding in patients with allergic rhinitis. N Engl J Med. 2003;349:237-46. [ Links ]
22. Woodcock A, Forster L, Matthews E, Martin J, Letley L, Vickers M, et al. Control of exposure to mite allergen and allergen-impermeable bed covers for adults with asthma. N Engl J Med 2003;349:225-36. [ Links ]
23. Chowdhury BA. Correspondence: bed covers and dust mites. N Engl J Med. 2003;349:1668-9. [ Links ]
24. Boner AL, Peroni D, Sette L, Valletta EA, Piacentini G. Effects of allergen exposure-avoidance on inflammation in asthmatic children. Allergy. 1993;48:119-123. [ Links ]
25. Platts-Mills TA, Tovey ER, Mitchell EB, Moszoro H, Nock P, Wilkins SR. Reduction of bronchial hyperactivity during prolonged allergen avoidance. Lancet. 1982;2:675-8. [ Links ]
26. Vervloet D, Penaud A, Razzouk H, Senft M, Arnaud A, Boutin C, et al. Altitude and house dust mites. J Allergy Clin Immunol. 1982;69:290-6. [ Links ]
27. Boner AL, Peroni DG, Piacentini GL, Venge P. Influence of allergen avoidance at high altitude on serum markers of eosinophil activation in children with allergic asthma. Clin Exp Allergy. 1993;23:1021-6. [ Links ]
28. Peroni DG, Boner AL, Vallone G, Antolini I, Warner JO. Effective allergen avoidance at high altitude reduces allergen-induced bronchial hyperresponsiveness. Am J Respir Crit Care Med. 1994;149:1442-6. [ Links ]
29. Sopelete MC, Silva DA, Arruda LK, Chapman MD, Taketomi EA. Dermatophagoides farinae (Der f 1) and Dermatophagoides pteronyssinus(Der p 1)allergen exposure among subjects living in Uberlandia, Brazil. Int Arch Allergy Immunol. 2000;122:257-63. [ Links ]
30. Terra SA, Silva DA, Sopelete MC, Mendes J, Sung SJ, Taketomi EA. Mite allergen levels and acarologic analysis in house dust samples in Uberaba,Brazil. J Investig Allergol Clin Immunol. 2004;14:232-7. [ Links ]
31. Simplicio EC, Silva DA, Braga IA, Sopelete MC, Sung SJ, Taketomi EA. Mite and pet allergen exposure in hotels in Uberlândia, Midwestern Brazil. Indoor Air. 2007 (in press). [ Links ]
32. Silva MC, Justino CM, Pereira FL, Segundo GR, Silva DA, Sung-Sang JS, et al. Exposição alergênica em cinemas na cidade de Goiânia, GO. Rev Bras Alergia Imunopatol. 2005;28:194-7. [ Links ]
33. Pereira FL, Silva DA, Sopelete MC, Sung SS, Taketomi EA. Mite and cat allergen exposure in Brazilian public transport vehicles. Ann Allergy Asthma Immunol. 2004;93:179-84. [ Links ]
34. Justino CM, Segundo GR, Pereira FL, Silva DA, Sopelete MC, Sung SS, et al. Mite and pet allergen exposure in Brazilian private cars. Ann Allergy Asthma Immunol. 2005;94:658-61. [ Links ]
35. Justino CM, Pereira FL, Segundo GR, Sopelete MC, Silva DA, Sung SJ, et al. Exposição alergênica em veículos privados de passeio e transporte escolar em Uberlândia, MG. Rev Bras Alergia Imunopatol. 2005;28:94-8. [ Links ]
36. Taketomi EA, Justino CM, Pereira FL, Segundo GR, Sopelete MC, Sung SJ, et al. Taxis but not private cars are mite allergen reservoirs in Brazil. J Investig Allergol Clin Immunol. 2006;16(1):34-6. [ Links ]
37. Luczynska CM, Arruda LK, Platts-Mills TA, Miller JD, Lopez M, Chapman MD. A two-site monoclonal antibody ELISA for the quantification of the major Dermatophagoides spp. allergens, Der p I and Der f I. J Immunol Methods. 1989;118:227-35. [ Links ]
38. Lowe LA, Woodcock A, Murray CS, Morris J, Simpson A, Custovic A. Lung function at age 3 years: effect of pet ownership and exposure to indoor allergens. Arch Pediatr Adolesc Med. 2004;158:996-1001. [ Links ]
39. Rullo VE, Rizzo MC, Arruda LK, Sole D, Naspitz CK. Daycare centers and schools as sources of exposure to mites, cockroach, and endotoxin in the city of Sao Paulo, Brazil. J Allergy Clin Immunol. 2002;110:582-8. [ Links ]
40. Perfetti L, Ferrari M, Galdi E, Pozzi V, Cottica D, Grignani E, et al. House dust mites (Der p 1, Der f 1), cat (Fel d 1) and cockroach (Bla g 2) allergens in indoor work-places (offices and archives). Sci Total Environ. 2004;328:15-21. [ Links ]
41. Custovic A et al. Domestic allergens in public places III: house dust mite, cat, dog and cockroach allergens in British hospitals. Clin Exp Allergy. 1998: 28, 53-59. [ Links ]
42. Custovic A, Taggart SC, Woodcock A. House dust mite and cat allergen in different indoor environments. Clin Exp Allergy. 1994;24:1164-8. [ Links ]
43. Wickens K, Martin I, Pearce N, Fitzharris P, Kent R, Holbrook N, et al. House dust mite allergen levels in public places in New Zealand. J Allergy Clin Immunol. 1997;99:587-93. [ Links ]
Address for correspondence: This paper was submitted to the RBORL-SGP (Publishing Manager System) on 1 March 2007. code 3712.
Ernesto Akio Taketomi
Avenida Pará 1720 Bloco 4C Campus Umuarama
Laboratório de Alergia e Imunologia Clínica Universidade Federal de Uberlândia
Uberlândia MG 38400-902
CAPES, CNPq, FAPEMIG
The article was accepted on 14 April 2007.
Address for correspondence:
This paper was submitted to the RBORL-SGP (Publishing Manager System) on 1 March 2007. code 3712.