1. The New England Journal of Medicine Impact Factor: 50.017 Rank: 1/107 (Medicine, General & Internal) Environmental Exposure to Endotoxin and It’s Relation to Asthma in School-age Children INSTRUCTOR Conrad (Dan) Volz, DrPH, MPH Mark WONG CHEN

2. Background

3. Endotoxin - The prototypical examples of endotoxin are lipopolysaccharide (LPS) found in the outer membrane of various Gram-negative bacteria and are an important component of their ability to cause disease. Allergen - An allergen is a nonparasitic antigen capable of stimulating some individuals mount an IgE response against common environmental antigens. December 6, 2010 2

4. Introduction

5. Asthma is the most common chronic disease and accounts for substantial morbidity and health care cost. Various environment factors have been thought to play key parts in development of allergies the causes of this disease remain unclear (D’Amato G et al. 2000)(Gold DR et al 2000) Improvements in public health and hygiene may increase the predisposition to chronic allergic conditions during childhood (Strachan DP et al. 2000) December 6, 2010 4

6. The microbial substances are recognized by the innate immune sys- tem in the absence of overt infection, and they induce a potent inflammatory response (Delves PJ et al. 2000) Environmental exposure to microbial products may have a crucial role during the maturation of a child’s immune response, causing the development of tolerance to other components December 6, 2010 5

7. They investigated the relation between exposure to microbial products and the occurrence of childhood asthma and allergies. December 6, 2010 6

8. Methods

9. Study Population There were 812 children aged 6 to 13. Located in rural areas of Austria, Germany, and Switzerland Farming and non-farming family December 6, 2010 8

10. Dust Sampling Vacuuming each mattress for2 minutes/m 2 The material obtained was divided in two for measurement of endotoxin and allergen content All field workers were centrally trained and certified to ensure similarity of sampling. December 6, 2010 9

11. Measurements of Endotoxin Levels Dust sample was stored at room temperature and shipped within one week after collection to the central laboratory Endotoxin results were expressed as endotoxin units per milligram of dust and as endotoxin units per square meter of surface area of the sampled mattress (Hollander et al. 1993) December 6, 2010 10

12. Measurements of Allergen Levels in Dust Samples The second dust sample was frozen at -20°C for at least two days and then shipped to one central laboratory The sample are stored at 4°C un til it was analyzed for  D. pteronyssinus (Der p1 protein) ‚ D. farinae (Der f1 protein) ƒ Felis domesticus (Fel d1 protein) December 6, 2010 11

13. Questionnaire and Interview The prevalence of diseases and symptoms and potential explanatory and confounding factors were assessed by a questionnaire given to the parents that included the questions of the International Study of Asthma and Allergies in Childhood December 6, 2010 12

14. Testing for Specific IgE in Serumc The level of specific IgE against airborne allergens in all serum samples was measured by fluorescence enzyme immunoassay. Atopy was defined by at least one positive test for specific IgE indicating a titer of at least 3.5 kU/ L December 6, 2010 13

15. Assessment of Cytokine Production by Peripheral-Blood Leukocytes Cells were stimulated either with 10 μg of lipopolysaccharide per milliliter for 24 hours or with staphylococcal enterotoxin B for 72 hours 下 TNF-α IFN-γ Interleukin-10 Interleukin-12 December 6, 2010 14

16. Results

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23. Discussion

24. The mechanisms by which endotoxin exposure may protect against the development of atopic immune responses and diseases are not fully understood. Although only current endotoxin exposure was measured, the levels are likely to reflect long- term exposure. Therefore, long-term, high-level environmental exposure may favor a state of tolerance (Lauener R et al. 2002) December 6, 2010 23

25. The protective effect of endotoxin exposure at school age was observed only for atopic wheeze and asthma, not for nonatopic wheeze. Children come into close contact with the microbial environment of their beds while sleeping and since the reproducibility of repeated endotoxin measurements is greater for dust from the bed than for dust from the floor (Fearon DT et al. 1996) December 6, 2010 24

26. Limitations

27. The study population focus on children, but did not observe their health conditions continuously. Target only focuses on German, other race was not included in the author’s studies. The authors operate their studies only in rural area, I think should operate same study in urban area. December 6, 2010 26

28. Limitations They did not assess other bacterial components, such as nonmethylated cytidine phosphate guanosine dinucleotides specific for prokaryotic DNA (CpG motifs) or cell-wall components from atypical mycobacteria or gram-positive bacteria, which are known to affect immune responses in ways similar to that of endotoxin. December 6, 2010 27

29. Conclusion

30. Mechanisms relating to the recognition of the microbial compounds by the innate immune system and the regulation of the resulting inflammatory responses through adaptive immunity are likely to be of key importance for the development of atopic illnesses such as hay fever and childhood asthma and wheeze. December 6, 2010 29

31. Thank ‘s for your listening