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Changes in immune cell composition during the immunological window within the first years after birth are not fully understood, especially the effect that different lifestyles might have on immune cell functionality.Peripheral blood mononuclear cells from mothers and their children at birth and at two anvd five years were analyzed by mass cytometry. Immune cell composition and functionality was analyzed according to family lifestyle (anthroposophic and non-anthroposophic).We found no significant differences in the proportions of major immune lineages between anthroposophic and non-anthroposophic children at each time point, but there were clear changes over time in the proportions of mononuclear leukocytes, especially in B-cells and T lymphocytes. Phenotypic distances between cord blood and maternal blood were high at birth but decreased sharply the first two years, indicating strong phenotypic convergence with maternal cells. We found that children exhibited similar stimulation responses at birth, but subsequently segregated into two discrete functional trajectories. Trajectory 1 was associated with a decrease in tumor necrosis factor alpha (TNFa) production by CD4This exploratory study suggests that leukocyte frequencies and cell phenotypes change with age in the same way across all children, while functional development follows one of two discrete trajectories that largely segregate by family lifestyle, supporting the hypothesis that early environmental exposures imprint immune cell function which may contribute to IgE sensitization. Our results also support that the first two years are critical for the environmental exposures to imprint the immune cells. Further studies with larger sample sizes are required to validate our findings.

Purpose Little is known about the longitudinal development of different plasma protein levels during early childhood and particularly in relation to lifestyle factors. This study aimed to monitor the plasma proteome early in life and the influence of different lifestyles. Experimental design A multiplex bead-based immunoassay was used to analyze plasma levels of 97 proteins in 280 blood samples longitudinally collected in children at 6, 12, 24 and 60 months of age living in families with an anthroposophic (n = 15), partly anthroposophic (n = 27) or non-anthroposophic (n = 28) lifestyle. Results A total of 68 proteins (70%) showed significantly altered plasma levels between 6 months and 5 years of age. In lifestyle stratified analysis, 59 of 97 (61%) proteins were altered over time within one or more of the three lifestyle groups. Nearly half of these proteins (28 out of 59) changed irrespective of lifestyle. The temporal changes represented four longitudinal trends of the plasma proteins during development, also following stratification of lifestyle. Conclusions and clinical relevance Our findings contribute to understand the development of the plasma proteome under the influence of lifestyle exposures in early childhood. This article is protected by copyright. All rights reserved.

Background: The increasing incidence of allergic diseases highlights the importance of finding underlying mechanisms. Early vaccination has been suggested as one influential factor. However, it is difficult to find a study group with a large variation between subjects concerning compliance to the official vaccination program. The anthroposophic lifestyle is of interest in this context. Moreover, cohort studies show that children of families with this lifestyle run a lower risk of allergic sensitization and allergy-related disease. Methods: From the prospective birth cohort ALADDIN we included one group from the anthroposophic community, with restrictive attitudes concerning vaccinations, and two other groups of age-matched children with more conventional parental lifestyles. In all, 466 children were followed from birth to five years of age. Detailed vaccination data and blood samples were collected at six months, one, two, and five years. Information was also obtained on risk factors for allergy. The outcome variable, allergic sensitization was defined as allergen-specific serum IgE levels ≥ 0.35 kUA/L. Findings: In a logistic regression model adjusted for socio-demographics and established allergy risk factors, vaccination at later age or having a lower number of injections or vaccines were associated with low OR for allergic sensitization during the first year of life. However, after adjustment for anthroposophic lifestyle, no statistically significant associations remained. The adjusted OR for sensitization at five years of age in children not receiving any vaccinations (n = 54) was 0.98 [95% CI 0.38–2.57]. Interpretation: We found no support for an association between early childhood vaccination and subsequent allergic sensitization. Our findings do not support scepticism towards early childhood vaccination motivated by allergy risk. Keywords: ALADDIN, Allergic sensitization, Anthroposophic lifestyle, Early childhood, Vaccination

BACKGROUND IgE sensitization is usually associated with allergy-related diseases, but may also occur in asymptomatic individuals. The clinical importance of IgE antibody concentrations in the interval 0.1-0.34 kU/L in early life in relation to allergy development is poorly evaluated. OBJECTIVE To assess the relevance of low specific IgE (s-IgE) to hen's egg, cow's milk and peanut at 6 months of age for development of sensitization and allergy-related disease during early childhood. METHODS s-IgE concentrations to relevant allergens from blood samples taken at 6 months and 1, 2 and 5 years from children in the prospective ALADDIN cohort were divided into three categories: non-sensitized (

Immunization is highly effective in preventing infectious diseases and therefore an indispensable public health measure. Allergic patients deserve access to the same publicly recommended immunizations as nonallergic patients unless risks associated with vaccination outweigh the gains. Whereas the number of reported possible allergic reactions to vaccines is high, confirmed vaccine-triggered allergic reactions are rare. Anaphylaxis following vaccination is rare, affecting less than 1/100,000, but can occur in any patient. Some patient groups, notably those with a previous allergic reaction to a vaccine or its components, are at heightened risk of allergic reaction and require special precautions. Allergic reactions, however, may occur in patients without known risk factors and cannot be predicted by currently available tools. Unwarranted fear and uncertainty can result in incomplete vaccination coverage for children and adults with or without allergy. In addition to concerns about an allergic reaction to the vaccine itself, there is fear that routine childhood immunization may promote the development of allergic sensitization and disease. Thus, although there is no evidence that routine childhood immunization increases the risk of allergy development, such risks need to be discussed. This article is protected by copyright. All rights reserved.

Background & Aims Little is known about the natural history of childhood recurrent abdominal pain (RAP). We investigated the prevalence and progression of childhood RAP and its association with Rome III abdominal pain–related functional gastrointestinal disorders (AP-FGID) and irritable bowel syndrome (IBS) during adolescence. Methods We collected data from a prospective population-based birth cohort study of 4089 children, born from 1994 through 1996 in Sweden. We analyzed data from 2455 children with complete follow-up evaluation at ages 1, 2, 12, and 16 years and no parent-reported diagnoses of inflammatory bowel diseases or celiac disease at ages 12 or 16 years. A subpopulation of 2374 children who had answered questions based on the Rome III criteria at age 16 years was identified. We assessed RAP at 3 assessment points and defined it as parent-reported attacks of colic in early childhood (1–2 years) and as self-reported weekly abdominal pain at ages 12 years and 16 years. AP-FGID at age 16 years was defined according to the Rome III criteria. Results RAP was reported by 26.2% of children on at least 1 of 3 assessment points, of which 11.3% reported symptoms more than once. Children with RAP at 12 years had persistent symptoms at 16 years in 44.9% of cases and increased risks for RAP (relative risk, 2.2; 95% CI, 1.7–2.8), any AP-FGID (relative risk, 2.6; 95% CI, 1.9–3.6), and IBS (relative risk, 3.2; 95% CI, 2.0–5.1) at 16 years. Early childhood RAP was not associated significantly with any outcome. Conclusions RAP affects many children from early childhood through age 16 years, but most children do not have persistent symptoms throughout childhood. RAP at age 12 years is a risk factor for RAP, any Rome III AP-FGID, and IBS, at age 16 years.

BACKGROUND: Immunomodulatory effects of sublingual immunotherapy on systemic and mucosal mediators in allergic children are largely unexplored. The aim of this study was to investigate allergy-related cytokine and chemokine levels, as well as IgA-responses upon a 3-year treatment with timothy grass pollen sublingual immunotherapy in children with allergic rhinoconjunctivitis. METHODS: From children included in the GRAZAX® Asthma Prevention study, blood and saliva samples were analyzed at inclusion, after 3 years of treatment, and 2 years after treatment ending. By means of Luminex and ELISA methodologies, allergy-related cytokines and chemokines were measured in plasma samples and allergen-stimulated peripheral blood mononuclear cell supernatants. Furthermore, studies of total, secretory, and Phl p 1-specific salivary IgA antibodies were performed using the same methods. RESULTS: GRAZAX® -treated children exhibited significantly higher levels of Phl p 1-specific salivary IgA and serum IgG4 , along with significantly lower skin prick test positivity, after 3 years of treatment and 2 years after treatment cessation. Additionally, plasma levels of the Th1-associated chemokines CXCL10 and CXCL11 were significantly higher in treated than untreated children at these time points. Timothy-induced ratios of IL-5/IL-13 over IFN-γ were significantly decreased after 3 years with active treatment, as were symptoms of allergic rhinitis in terms of both severity and visual analogue scale scores. However, no consistent correlations were found between the clinical outcomes and immunologic parameters. CONCLUSION: Phleum pratense sublingual immunotherapy in grass pollen allergic children modulates the immune response in the oral mucosa as well as systemically-by increasing Th1-responses, decreasing Th2-responses, and inducing immunoregulatory responses-all signs of tolerance induction. Funding agencies: Research Council for South-East Sweden; University Hospital of Linkoping, Sweden

Prenatal environmental exposures are considered to contribute to the development of allergic sensitization by epigenetic mechanisms. The role of histone acetylation in the placenta has not been examined yet. We hypothesized that placental histone acetylation at the promoter regions of allergy-related immune regulatory genes is associated with the development of sensitization to allergens in the child. Histones H3 and H4 acetylation at the promoter regions of 6 selected allergy-related immune regulatory genes was assessed by a chromatin immunoprecipitation assay in 173 term placentas collected in the prospective birth-cohort ALADDIN. The development of IgE sensitization to allergens in the children was followed from 6 months up to 5 years of age. We discovered significant associations of histone acetylation levels with decreased risk of allergic sensitization in 3 genes. Decreased risk of sensitization to food allergens was associated with higher H3 acetylation levels in placentas at the IFNG and SH2B3 genes, and for H4 acetylation in HDAC4. Higher HDAC4 H4 acetylation levels were also associated with a decreased risk of sensitization to aeroallergens. In conclusion, our results suggest that acetylation of histones in placenta has a potential to predict the development of sensitization to allergens in children.

The trajectory of IgE levels before, during and after pregnancy in sensitized individuals is characterized by significant increase in specific IgE to birch allergens but not to other allergens after multiple testing. This increase may warrant some surveillance in the antenatal care for those with clinical symptoms.

Background Breastmilk fatty acids (FAs) have been associated with childhood allergic disease. Children of families with an anthroposophic lifestyle have a low prevalence of sensitization compared to reference groups. This study aimed to investigate whether the lower prevalence of sensitization among these children can be explained by the differences in breastmilk FA composition. Methods The prospective birth cohort ALADDIN included 330 children from anthroposophic, partly anthroposophic and nonanthroposophic families recruited between 2004 and 2007 in Sweden. In total, 245 breastmilk samples, collected at 2 months of age, were analysed for FA composition. Allergen-specific IgE levels against seven common allergens were measured in the blood samples at the ages of 6, 12 and 24 months. Data were analysed longitudinally using generalized estimating equations. Results An inverse association was observed between total concentration of omega-3 PUFA in breastmilk and sensitization in the child up to 24 months of age (highest vs lowest quartile, RRadj 0. 49, 95% CI 0.23–1.05, P for trend 0.024). No associations were observed between omega-6 PUFAs or ruminant FAs and sensitization. Overall, we observed 56% lower risks of sensitization among the anthroposophic group compared to the nonanthroposophic group (RRadj 0.44, 95% CI 0.21–0.90). This association remained largely unchanged when breastmilk omega-3 PUFA was included in the model. Conclusion Our results indicate that a higher concentration of omega-3 PUFAs in breastmilk may be associated with a reduced risk of sensitization up to 24 months of age; however, this did not explain the lower risk of sensitization among children of anthroposophic families.

We construct an explicit and universal A-infinity deformation of Batalin-Vilkovisky algebras, with all coefficients expressed as rational sums of multiple zeta values. If the Batalin-Vilkovisky alg ...

Aim To investigate (i) whether maternal sensitisation to allergens, and lifestyle can influence the risk of acute and chronic inflammation of the placenta, in the forms of chorioamnionitis and villitis, respectively, and (ii) whether these placental inflammations are associated with the outcome of sensitisation for the child during preschool age. Methods Placentas from term uncomplicated pregnancies (n = 275) in the assessment of lifestyle and allergic disease during infancy study were analysed for the presence of acute chorioamnionitis and chronic villitis. Stepwise logistic regression was performed to estimate the relative risk of placental inflammation in relation to maternal allergic sensitisation and lifestyle, and the association between placental inflammation and sensitisation of the child up to five years of age. Results Parity and delivery at home were independently associated with chorioamnionitis, home delivery only with the low grade. Maternal allergic sensitisation was associated with increased risk of villitis in the bivariable model, however, not in the multivariable model. No significant associations were detected between placental inflammation and the outcome of sensitisation to allergens at five years of age. Conclusion Our data do not support the hypothesis that the increased risk for sensitisation of a child when the mother is allergic is mediated via placental inflammation.

BACKGROUND: Allergy immunotherapy targets the immunological cause of allergic rhinoconjunctivitis and allergic asthma and has the potential to alter the natural course of allergic disease.OBJECTIVE: The primary objective was to investigate the effect of the SQ grass sublingual immunotherapy tablet compared with placebo on the risk of developing asthma.METHODS: A total of 812 children (5-12 years), with a clinically relevant history of grass pollen allergic rhinoconjunctivitis and no medical history or signs of asthma, were included in the randomized, double-blind, placebo-controlled trial, comprising 3 years of treatment and 2 years of follow-up.RESULTS: There was no difference in time to onset of asthma, defined by prespecified asthma criteria relying on documented reversible impairment of lung function (primary endpoint). Treatment with the SQ grass sublingual immunotherapy tablet significantly reduced the risk of experiencing asthma symptoms or using asthma medication at the end of trial (odds ratio = 0.66, P < .036), during the 2-year posttreatment follow-up, and during the entire 5-year trial period. Also, grass allergic rhinoconjunctivitis symptoms were 22% to 30% reduced (P < .005 for all 5 years). At the end of the trial, the use of allergic rhinoconjunctivitis pharmacotherapy was significantly less (27% relative difference to placebo, P < .001). Total IgE, grass pollen-specific IgE, and skin prick test reactivity to grass pollen were all reduced compared to placebo.CONCLUSIONS: Treatment with the SQ grass sublingual immunotherapy tablet reduced the risk of experiencing asthma symptoms and using asthma medication, and had a positive, long-term clinical effect on rhinoconjunctivitis symptoms and medication use but did not show an effect on the time to onset of asthma.

Background Infants from anthroposophic families have low cortisol levels and low risk of IgE-sensitization during first 2 years of life. Our aim was to study the impact of an anthroposophic lifestyle and cortisol levels at 6 months on allergy sensitization up to age 5 years. Methods A total of 507 families participated from maternal healthcare centers. Parental lifestyle was categorized as anthroposophic, partly anthroposophic, or non-anthroposophic. Blood samples for analyzes of sensitization were obtained from parents at inclusion and from children at 6, 12, 24, and 60 months. Salivary samples were collected at home at 6 months. Results Sensitization increased from 2.9% to 26.0% in the anthroposophic group, from 8.4% to 26.8% in the partly anthroposophic group, and from 19.1% to 44.1% in the non-anthroposophic group. Children from anthroposophic families had lower cortisol levels in the morning, afternoon, and evening. The odds ratio (OR) for anthroposophic lifestyle was always

We study actions of the Grothendieck-Teichmuller group GRT on Poisson coho- mologies of Poisson manifolds, and prove some "go" and "no-go" theorems associated with these actions. Mathematics Subject Classification (2010). 53D55, 16E40, 18G55, 58A50.

OBJECTIVE: To identify morning salivary cortisol reference values in infancy and at 2 years of age and to investigate the influence of age, sex and acute bronchiolitis. STUDY DESIGN: In this South-East Norwegian cohort study, 308 children hospitalized with moderate to severe acute bronchiolitis in infancy in 2010-2011 were compared with 223 healthy controls included in 2012 by measuring morning salivary cortisol levels at inclusion and at 2 years of age. Samples were collected shortly after awakening after 6 am. The influences of age, sex, and acute bronchiolitis were assessed by regression analysis. RESULTS: In infancy, cortisol values were higher in acute bronchiolitis, with an age- and sex-adjusted weighted mean group difference of 13.9 nmol/L (95% CI 8.1-19.7; P

Long-term birth cohort studies are essential to understanding the life course and childhood predictors of allergy and the complex interplay between genes and the environment (including lifestyle and socioeconomic determinants). Over 100 cohorts focusing on asthma and allergy have been initiated in the world over the past 30 years. Since 2004, several research initiatives funded under the EU Framework Program for Research and Technological Development FP6-FP7 have attempted to identify, compare, and evaluate pooling data from existing European birth cohorts (GA2LEN: Global Allergy and European Network, FP6; ENRIECO: Environmental Health Risks in European Birth Cohorts, FP7; CHICOS: Developing a Child Cohort Research Strategy for Europe, FP7; MeDALL: Mechanisms of the Development of ALLergy, FP7). However, there is a general lack of knowledge about these initiatives and their potentials. The aim of this paper is to review current and past EU-funded projects in order to make a summary of their goals and achievements and to suggest future research needs of these European birth cohort networks.

Asthma, eczema, food allergy and allergic rhinitis are common and the cause is still largely unknown, but it is the subject of extensive research. The research results are inconclusive, probably due to individual differences in vulnerability to different exposures and to a highly multifactorial origin. Several environmental factors have been identified as risk factors or as having a potentially protective effect. Smoking during pregnancy or childhood increases the risk of obstructive symptoms and asthma. Breastfeeding reduces the risk of infectious diseases and thereby also the risk of early obstructive symptoms. Even if mothers avoid certain foods during pregnancy or breastfeeding, this does not reduce the risk of allergic disease in the child. The child should be introduced to foods between four and six months of age, without avoiding or delaying any particular food. Vaccinations do not affect the risk of allergic disease. The risk of allergic disease is not increased by exposure to furry animals in the home, if the child is healthy. However, children who are already allergic should at least avoid the animals to which they are allergic.

Background: Previous studies on anthroposophic lifestyle and allergy show that the children have less risk of developing allergies. All studies so far have been retrospective and have included children in school age. In view of the facts that this lifestyle seems to protect children from allergies and that different symptoms of atopy have increased dramatically during the last decades it is of general interest to study this group of children more in detail. The earlier findings have now been followed up in a prospective research program ALADDIN (Assessment of Life style and Allergic Disease During Infancy), applying different approaches. One of these is to focus on stress (as measured by cortisol) as a factor that may underlie the decreased risk of allergy in children from anthroposophic families. Aim: This thesis is based on data from the ALADDIN study focusing on influence of family lifestyle on allergy sensitization early in children’s life in relation to psychosocial factors and salivary cortisol as an indicator of stress.Methods: A total of 552 families were recruited during pregnancy or during the first month of the child´s life in two recruitment waves, 330 families between September 2004 and November 2007 and another 222 families between March 2008 and January 2011. They were recruited at anthroposophic maternal and child health care centers (MCHC) and from conventional MCHCs. Data on demographics and exposures were obtained by questionnaires and interviews. Parental capacity to adapt to stressors was evaluated by means of the questionnaire “Sense of Coherence” (SOC; Antonovsky). Salivary samples were collected at home from the infants and both parents for analyzes of cortisol. Blood samples were obtained from the parents and from the children for analyzes of IgE.Results: Many family related characteristics differed markedly between the groups before and during pregnancy, during delivery and the first 12 months of age. Children from anthroposophic families had lower levels of salivary cortisol compared to peers from families with a more conventional lifestyle on all sampling occasions at 6 months of age and on some of the occasions at 12 and 24 months of age. There were no differences concerning cortisol between parents with different lifestyle and no significant differences concerning SOC-scores between the three lifestyle groups. An anthroposophic lifestyle was associated with a lower risk of allergic sensitization up to five years of age. This risk was partially explained by lower cortisol levels during infancy. Children in families with a partly anthroposophic lifestyle also had substantially lower risk of sensitization.Conclusion: An anthroposophic lifestyle protects from development of allergy during childhood, at least up to five years of age. This protective capacity is partly mediated by low cortisol levels during infancy but is also dependent on unknown characteristics of this lifestyle. These results call for further studies on health related effects of an anthroposophic lifestyle.

Aim Abdominal pain of functional origin is very common in childhood, and environmental factors are thought to be of aetiologic importance. The anthroposophic lifestyle has dietary and lifestyle characteristics that may influence child health, and this study aimed to assess the effect of such lifestyles on abdominal pain of functional origin. Methods A prospective Swedish lifestyle cohort (n = 470) was followed from birth to five years of age. Family lifestyles were characterised through questionnaires. Abdominal pain was defined as irritable bowel syndrome or functional abdominal pain according to the Rome III criteria and measured with parental questionnaires and interviews at the age of five. Results The prevalence of abdominal pain was 15%. Children were more likely to have abdominal pain at five years of age if their family had a partly anthroposophic lifestyle, with an adjusted odds ratio (OR) of 2.61 (95% CI 1.15–5.93), or an anthroposophic lifestyle, with an adjusted OR of 2.34 (95% CI 0.96–5.70). Conclusion A family lifestyle with anthroposophic characteristics was associated with an increased risk of abdominal pain in five-year-old children. The mechanisms for this increase were unclear, but we speculate that there may have been different prerequisites for coping with stressors.

Pre- and postnatal stress have been related to allergy in children, but evidence from prospective studies is limited. Several environmental factors can influence the salivary cortisol level, which is used as a measure of activity of the hypothalamic-pituitary-adrenal axis.The aim of this study was to assess the association between salivary cortisol levels at 6 months of age and allergic manifestations during the first 2 years of life.Salivary samples for the analysis of cortisol level were collected at 6 months of age on 3 occasions during 1 day from 203 children. Blood samples were collected at 6, 12, and 24 months of age for analyses of specific IgE. Information on allergy-related symptoms was obtained by repeated examinations of the children. Generalized estimating equation statistics were used to calculate the overall risk for outcome measures.The adjusted odds ratio for the relationship between morning cortisol level and IgE sensitization was 1.60 (95% CI, 1.22-2.10, P = .001) and for eczema it was 1.28 (95% CI, 1.03-1.59, P = .026). The odds ratio for afternoon cortisol level in relation to sensitization and eczema was 1.56 (95% CI, 1.26-1.94, P .001) and 1.33 (95% CI, 1.12-1.58, P = .001), respectively, and for evening cortisol level it was 1.49 (95% CI, 1.22-1.83, P .001) and 1.37 (95% CI, 1.18-1.59, P .001). Salivary cortisol level in the evening was associated with food allergy.The association between salivary cortisol levels in infancy and allergic sensitization and allergic symptoms suggests a role of an altered hypothalamic-pituitary-adrenal axis in the etiological process of allergies.

Background: Several cross-sectional studies indicate that an anthroposophic lifestyle reduces the risk of allergy in children. We initiated the Assessment of Lifestyle and Allergic Disease During Infancy (ALADDIN) birth cohort to elucidate the role of specific factors supposed to mediate this effect. The aims of this study are to describe the ALADDIN cohort and to report patterns of exposure and allergic sensitization during the first years of life. Methods: The ALADDIN study is a prospective birth cohort study of 330 children from families with an anthroposophic, partly anthroposophic, or nonanthroposophic lifestyle. The children and their parents were following an extensive data collection scheme, including repeated questionnaires and biological samples. Blood samples were collected from the parents and from the child at birth as well as at 6, 12, and 24 months of age. Results: Several lifestyle factors differed between the groups, such as diet, medication, and place of delivery. Children of families with an anthroposophic lifestyle had a markedly decreased risk of sensitization during the first 2 years of life compared with children of nonanthroposophic families with adjusted OR 0.25 (95% CI 0.10–0.64) and P-value 0.004. A similar situation held true for children from families with a partly anthroposophic lifestyle, adjusted OR 0.31 (95% CI 0.15–0.54), and P-value 0.002. Conclusions: The anthroposophic lifestyle comprises several factors of interest for allergy development and is here shown to be associated with reduced risk of IgE sensitization already in infancy. Identifying the factors responsible for this association would be of significant clinical importance.

The anthroposophic lifestyle implies environmental conditions for the infant aimed at reducing negative stress stimulation and is also related to a lower prevalence of allergic diseases in children. One aim of this prospective birth cohort study was to assess stress levels in infants with an anthroposophic lifestyle. A total of 330 infants from families with anthroposophic or more conventional lifestyles were followed from pregnancy of their mothers until the age of 6 months. Information on lifestyle factors was obtained from questionnaires. Salivary samples from 210 6-month olds and their parents were collected on three occasions during 1 day for analysis of cortisol. Infants from families with an anthroposophic lifestyle had significantly lower cortisol levels on all three sampling occasions compared to other infants. In the morning, the geometric means of salivary cortisol concentration (with 95% confidence limits) were 8.8 nmol/l (6.7-11.5), 11.3 nmol/l (9.3-13.7) and 14.9 nmol/l (11.3-19.6) in infants classified as anthroposophic, partly anthroposophic and non-anthroposophic, respectively (p=0.018). On the other hand, there was no difference in cortisol levels between the parents in the different groups. Several lifestyle factors differed significantly between the groups, but none of them independently explained the difference in cortisol levels. However, living on a farm during pregnancy was significantly associated with low saliva cortisol level in the infant. It can be concluded that low salivary cortisol levels in infants from anthroposophic families may be related to an environment with a lower degree of exposure to stress, which could influence the development of allergic diseases.

Background The evidence from previous studies on beneficial effects of breast-feeding in relation to development of asthma is conflicting. Objective To investigate the relation between breast-feeding and asthma and/or sensitization during the first 8 years of life. Method In a birth cohort, children were followed up to 8 years by questionnaires at ages 2 months and 1, 2, 4, and 8 years to collect information on exposures and health effects. Determination of serum IgE antibodies to common inhalant and food allergens was performed at 4 and 8 years. Longitudinal analyses were applied by using general estimated equations. The study population consisted of 3825 children (93% of the original cohort), of whom 2370 gave blood and 2564 performed lung function measurements at 8 years. Results Children exclusively breast-fed 4 months or more had a reduced risk of asthma during the first 8 years of life (adjusted odds ratio [OR], 0.63; 95% CI, 0.50-0.78) compared with children breast-fed less than 4 months. At 8 years, reduced risks of sensitization (adjusted OR, 0.79; 95% CI, 0.64-0.99) and asthma in combination with sensitization (adjusted OR, 0.59; 95% CI, 0.37-0.93) were seen among children exclusively breast-fed 4 months or more. This group also had a significantly better lung function measured with peak expiratory flow. Conclusion Breast-feeding for 4 months or more seems to reduce the risk of asthma up to 8 years. At this age, a reduced risk was observed particularly for asthma combined with sensitization. Furthermore, breast-feeding seems to have a beneficial effect on lung function.

To cite this article: Joerink M, Oortveld MAW, Stenius F, Rindsjo E, Alm J, Scheynius A. Lifestyle and parental allergen sensitization are reflected in the intrauterine environment at gene expression level. Allergy 2010; 65: 1282–1289. Abstract Background: Environmental factors, including the intrauterine environment, can influence the risk of allergy development. In the present study, we investigated whether lifestyle and parental allergen sensitization status are reflected at gene expression level in the intrauterine environment. Methods: mRNA expression of 17 genes was determined by means of quantitative real-time PCR in term placenta of 36 families participating in the ALADDIN study (Assessment of Lifestyle and Allergic Disease During Infancy). Data were analysed using a linear regression model to estimate the influence of lifestyle and parental allergen sensitization on the relative mRNA expression levels. Immunohistochemistry on placenta biopsies was used to verify protein expression. Results: Significant differences in mRNA expression levels were detected at the foetal side of the placenta, where CD14 was expressed at higher levels in placentas from families living on a farm compared to not living on a farm, and IL-12(p40) was expressed at lower levels when the father was sensitized compared to nonsensitized. At the maternal side of the placenta, higher expression of STAT4 and lower expression of GATA3 were detected in families with sensitized compared to nonsensitized mothers, and IL-12(p40) was lower expressed when the families were living on a farm compared to not living on a farm. Immunohistochemistry performed for STAT4 and GATA3 showed that protein and mRNA levels correlated well. Conclusion: Living on a farm and parental allergen sensitization are reflected in the intrauterine environment at the gene expression level.

Background Childhood exposure to a farm environment has been shown to protect against the development of inflammatory diseases, such as allergy, asthma, and inflammatory bowel disease. Objective We sought to investigate whether both exposure to microbes and exposure to structures of nonmicrobial origin, such as the sialic acid N-glycolylneuraminic acid (Neu5Gc), might play a significant role. Methods Exposure to Neu5Gc was evaluated by quantifying anti-Neu5Gc antibody levels in sera of children enrolled in 2 farm studies: the Prevention of Allergy Risk factors for Sensitization in Children Related to Farming and Anthroposophic Lifestyle (PARSIFAL) study (n = 299) and the Protection Against Allergy Study in Rural Environments (PASTURE) birth cohort (cord blood [n = 836], 1 year [n = 734], 4.5 years [n = 700], and 6 years [n = 728]), and we associated them with asthma and wheeze. The effect of Neu5Gc was examined in murine airway inflammation and colitis models, and the role of Neu5Gc in regulating immune activation was assessed based on helper T-cell and regulatory T-cell activation in mice. Results In children anti-Neu5Gc IgG levels correlated positively with living on a farm and increased peripheral blood forkhead box protein 3 expression and correlated inversely with wheezing and asthma in nonatopic subjects. Exposure to Neu5Gc in mice resulted in reduced airway hyperresponsiveness and inflammatory cell recruitment to the lung. Furthermore, Neu5Gc administration to mice reduced the severity of a colitis model. Mechanistically, we found that Neu5Gc exposure reduced IL-17+ T-cell numbers and supported differentiation of regulatory T cells. Conclusions In addition to microbial exposure, increased exposure to non–microbial-derived Neu5Gc might contribute to the protective effects associated with the farm environment., published version, peerReviewed

Background: Immunoglobulin E (IgE) has been identified on macrophage-like cells in the villi of human placenta, irrespective of the serum IgE levels or allergy status of the mother. The origin of placental IgE is debated and it is not known if it is spontaneously produced, so-called ‘natural IgE’, or if it has any specificity for certain allergens. The aim of this study was to investigate if placental IgE originates from mother or child and to analyse its specificity. Methods: Immunoglobulin E was eluted from placenta by lowering the pH. Total and allergen-specific IgEs were measured in placenta eluate, maternal and cord blood plasma by means of ImmunoCAP (Phadia AB). The levels of natural antibodies were determined with an anti-phosphorylcholine (PC) enzyme-linked immunosorbent assay, as natural IgE has been shown in one previous publication with this assay. Results: Detectable amounts of IgE were eluted from 11/12 full-term placentas. Natural (anti-PC) IgE antibodies were detected in low amounts in maternal plasma but not in the placental eluate or in cord blood plasma. There was a significant correlation between the amount of total IgE eluted from placenta and the levels of total IgE in maternal plasma; however, not between maternal and cord blood plasma. Allergen-specific IgE was only found in placental eluates from mothers with specific IgE towards these allergens. Furthermore, there was a significant correlation between the amount of allergen-specific IgE eluted from placenta and the levels of allergen-specific IgE in maternal plasma. Allergen-specific IgE could not be detected in cord blood. Conclusion: These results suggest a maternal origin of placental IgE, which can be allergen-specific.

In this population-based study, 90 children from three European countries were examined to determine the impact of lifestyle on the fecal microbiota. The study was designed to assess the impact of two extreme lifestyles that we hypothesized could impact the microbial composition in the gut: i.e., an anthroposophic lifestyle (restricted use of antibiotics, greater consumption of fermented vegetables, etc.) versus living on a farm (greater consumption of farm milk, contact with animals, etc.). In previous studies, these lifestyles correlated with lower prevalence of allergies. Terminal restriction fragment length polymorphism (T-RFLP) was used to assess the bacterial composition in fecal samples since recent studies have shown that the majority of this community cannot be cultivated. The T-RFLP data were used to calculate richness and evenness of the fecal microbiota. Children that were attending Steiner schools (anthroposophic children) had a significantly higher diversity of microbes in their feces than farm children, who in turn also had lower diversity than the control groups. Specific primers were also used to focus on the Lactobacillus -like community (lactic acid bacteria [LAB]). Large differences were found in the LAB subpopulations in the sampled groups. In some children, the LAB subpopulation was dominated by a species that has not yet been cultivated.

Vernix caseosa (VC) is a protective layer that covers the skin of most human newborns. This study characterized the VC lipid mediator profile and examined its relationship to gestational period, gender of the newborn and maternal lifestyle. VC collected at birth from 156 newborns within the ALADDIN birth cohort was analyzed and 3 different groups of lipid mediators (eicosanoids and related oxylipin analogs, endocannabinoids and sphingolipids) were screened using LC-MS/MS. A total of 54 compounds were detected in VC. A number of associations between lipid mediators and the gestational period were observed, including increases in the ceramide to sphingomyelin ratio as well as the endocannabinoids anandamide and 2-arachidonoylglycerol. Gender-specific differences in lipid mediator levels were observed for all 3 lipid classes. In addition, levels of the linoleic acid oxidation products 9(10)-epoxy-12Z-octadecenoic and 12(13)-epoxy-9Z-octadecenoic acid (EpOMEs) as well as 12,13-dihydroxy-9Z-octadecenoic acid (DiHOME) were increased in VC of children from mothers with an anthroposophic lifestyle. Accordingly, VC was found to be rich in multiple classes of bioactive lipid mediators, which evidence lifestyle, gender and gestational week dependencies. Levels of lipid mediators in VC may therefore be useful as early stage non-invasive markers of the development of the skin as a protective barrier.

Francis Brown introduced a partial compactification $M_{0,n}^\delta$ of the moduli space $M_{0,n}$. We prove that the gravity cooperad, given by the degree-shifted cohomologies of the spaces $M_{0,n}$, is cofree as a nonsymmetric anticyclic cooperad; moreover, the cogenerators are given by the cohomology groups of $M_{0,n}^\delta$. This says in particular that $H^\bullet(M_{0,n}^\delta)$ injects into $H^\bullet(M_{0,n})$. As part of the proof we construct an explicit diagrammatically defined basis of $H^\bullet(M_{0,n})$ which is compatible with cooperadic cocomposition, and such that a subset forms a basis of $H^\bullet(M_{0,n}^\delta)$. We show that our results are equivalent to the claim that $H^k(M_{0,n}^\delta)$ has a pure Hodge structure of weight $2k$ for all $k$, and we conclude our paper by giving an independent and completely different proof of this fact. The latter proof uses a new and explicit iterative construction of $M_{0,n}^\delta$ from $\mathbb{A}^{n-3}$ by blow-ups and removing divisors, analogous to Kapranov's and Keel's constructions of $\overline M_{0,n}$ from $\mathbb{P}^{n-3}$ and $(\mathbb{P}^1)^{n-3}$, respectively., Comment: 39 pages. v2: significant revision, to appear in the Annales Scientifiques de l'ENS

Increased mRNA expression of glucocorticoid receptor-P in placenta is associated with a decreased risk of allergen sensitisation in the child

The intestinal flora is considered to have an impact on the development of the immune system. In the anthroposophic lifestyle, a diet comprising vegetables spontaneously fermented by lactobacilli, and a restrictive use of antibiotics, anti-pyretics and vaccinations, is typical. The aim of this study was to assess the gut flora in infants in relation to certain lifestyle characteristics associated with anthroposophy. Sixty-nine children < 2 years of age with an anthroposophic lifestyle, and 59 infants of a similar age with a traditional lifestyle, were clinically examined and questionnaire replies assessed. Fecal samples were analyzed by bacterial enumeration, bacterial typing through biochemical fingerprinting and by measuring microflora-associated characteristics (MACs). The numbers of colony-forming units (CFU)/g of feces were significantly higher for enterococci and lactic acid bacteria in children who had never been exposed to antibiotics (5.5 x 10 7 vs. 2.1 x 10 7 ; p < 0.001 and 10 × 10 7 vs. 4.1 x 10 7 ; p < 0.01, respectively). Furthermore, the number of enterococci was significantly higher in breastfed and vegetarian infants (p < 0.01). The diversity (Simpson's diversity index) of lactobacilli, as determined by biochemical fingerprinting, was higher in infants born at home than in those born in hospital (p < 0.01). Several MACs were related to specific lifestyle features, and infants with an anthroposophic lifestyle had a higher proportion of acetic acid and a lower proportion of propionic acid in their stool as compared to the control children. In conclusion, lifestyle factors related to the anthroposophic way of life influenced the composition of the gut flora in the infants. These differences may contribute to the lower prevalence of atopic disease previously observed in children in anthroposophic families.

An anthroposophic lifestyle, which has been associated with reduced allergy risk in children, has several characteristics that could influence gut microbiota. This study aimed to investigate the impact of anthroposophic lifestyle as well as specific early life exposures on the gut microbiota. In total, 665 stool samples from 128 mother-infant pairs from the ALADDIN birth cohort study were included. Samples collected from infants at ages 6 days, 3 weeks, 2 months and 6 months, and from their mothers before and after delivery, respectively, were analyzed using 454-pyrosequencing. Information regarding lifestyle exposures was collected prospectively through interviews and questionnaires. Six-month-old infants in anthroposophic families had a significantly higher abundance of Bifidobacterium and lower abundances of Bacteroides and Veillonella. Caesarean section and breastfeeding had a significant impact on the microbiota: caesarean section was primarily associated with delayed colonization of Bifidobacterium and Bacteroides, whereas breastfed children had a higher relative abundance of Bifidobacterium and a lower abundance of Clostridiales. However, despite large differences in lifestyle exposures, we determined no significant differences in the gut microbiota between the anthroposophic and non-anthroposophic mothers or their infants' before 6 months of age.

Allergic diseases develop in genetically susceptible individuals in a complex interplay with the environment, usually early in life. We have previously shown that the anthroposophic lifestyle is associated with reduced risk of allergic disease in children, but details on the influencing environmental factors are largely unknown. This study aims to elucidate if anthroposophic lifestyle influences fetal exposure to selected toxic and essential elements. Randomly selected non-smoking mothers with (n=40) and without (n=40) anthroposophic lifestyle from the prospective birth cohort ALADDIN were included. Concentrations of 12 toxic and essential elements were analyzed in full term placentas and in the erythrocyte fractions of maternal peripheral blood and of umbilical cord blood, using inductively coupled plasma mass spectrometry. Cadmium concentrations in maternal blood and placenta were significantly higher in mothers with an anthroposophic lifestyle (p

Background Breast-feeding has many beneficial effects on the developing immune system of the newborn. Breast milk contains immunoregulatory factors, such as nano-sized vesicles named exosomes. This study aimed at characterizing breast milk exosomes from human early milk and mature milk and to investigate whether allergic sensitization and an anthroposophic lifestyle could influence the exosome profile. Methods Breast milk was collected from 22 mothers at day 3–8 and from 61 mothers at 2 months postpartum, all part of the ALADDIN birth cohort. Isolated exosomes were captured on anti-MHC-class II- or anti-CD63 beads and analyzed by flow cytometry. Exosomal phenotype was related to lifestyle and allergic sensitization of the mothers, and sensitization of the child at 2 years of age. Results We found a higher content of exosomes in early milk compared with mature milk. Early milk exosomes were enriched in HLA-DR molecules and displayed significantly lower levels of HLA-ABC compared with those in mature milk. Phenotypically different subpopulations of exosomes were found in mature milk. Significantly lower levels of MUC1 were detected on CD63-enriched exosomes from sensitized mothers compared with nonsensitized. Furthermore, women with an anthroposophic lifestyle had significantly lower MUC1 expression on their HLA-DR-enriched milk exosomes and up-regulated levels of CD63 on CD63-enriched exosomes compared with nonanthroposophic mothers. Notably, mothers whose children developed sensitization had an increased amount of HLA-ABC on their milk exosomes enriched for CD63. Conclusions The phenotype of exosomes in breast milk varies with maternal sensitization and lifestyle, which might influence allergy development in the child.

To cite this article: Holm T, Rutishauser D, Kai-Larsen Y, Lyutvinskiy Y, Stenius F, Zubarev RA, Agerberth B, Alm J, Scheynius A. Protein biomarkers in vernix with potential to predict the development of atopic eczema in early childhood. Allergy 2014; 69: 104–112. The prevalence of AE has rapidly increased and is estimated to 15–30% in children and 2–10% in adults with the highest prevalence found in Northern Europe 1. The pathogenesis of AE is likely to result from the combination of a disturbed skin barrier and inappropriate immune responses with contributions from both genetic and environmental factors. Atopic eczema is often the first symptom early in life of allergic disorders that many times progress to food allergy, asthma, and allergic rhinitis, a phenomenon known as the atopic march 2,3. A recent report highlighted the need to identify different early biomarkers in AE with the goal to stop or even reverse the atopic march 4. Inverse relation has been found between the number of anthroposophic lifestyle characteristics and the risk of atopy in school children of families with an anthroposophic lifestyle 5,6. Anthroposophic lifestyle includes biodynamic food containing live lactobacilli and restrictive use of antibiotics, antipyretics, and vaccinations 7. To investigate how lifestyle factors influence the development of allergic disease early in life, the birth cohort designated Assessment of Lifestyle and Allergic Disease During INfancy (ALADDIN) was initiated 7. In this birth cohort, the children born into families with an anthroposophic or nonanthroposophic life style have been followed prospectively from birth and many different samples have been collected, among those vernix caseosa (vernix). Vernix is a yellow-white, cheese-like, material consisting of sebum, lanugo hairs, and desquamated squamous cells, which covers the neonate's skin during the last trimester of gestation 8,9. No other animal species produces vernix, making this material a unique human skin barrier film. It is believed to exhibit waterproofing, antioxidant, temperature-regulating, and anti-infective functions 8. The composition of vernix is mainly water (80.5%), lipids (10.3%), and proteins (9.1%) 9. Vernix is produced by the sebaceous glands, and in addition, lanugo hairs and desquamated squamous cells contribute to the formation of vernix 8. Thus, the origin of proteins found in vernix is to a large part from the fetal epidermis. In this study, we investigated whether protein biomarkers in vernix, which reflects the composition of the infant's epidermal barrier, can predict later development of AE. As a source of biomarkers, vernix is attractive because it is usually abundant and its collection is noninvasive. We also addressed whether lifestyle can influence the protein composition of vernix by further dividing the AE and healthy control groups into three lifestyle categories: anthroposophic, partly anthroposophic, and nonanthroposophic.

Background The etiology of recurrent abdominal pain of functional origin (AP) is largely unknown. Antibiotic treatment influences the intestinal microbiota, and a few studies have indicated an increased risk of AP in adults after antibiotic treatment. Corresponding data in children are lacking. The aim of this study was to explore the association between antibiotic treatment during childhood and AP at 12 years. Methods Two thousand seven hundred and thirty-two children from a Swedish, population-based birth cohort. Parents reported antibiotic use for the children between birth and 2 years. Antibiotic use between 9 and 12 years was collected from the Swedish Prescribed Drug Register. The children answered questionnaires regarding AP at age 12. We used logistic regression to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for AP at 12 years as a function of antibiotic use. Key Results Antibiotic treatment between 9 and 12 years was not associated with AP at 12. Children who had received ≥3 courses, or broad-spectrum antibiotics between 9 and 12 years had an increased risk of AP at 12, but these associations failed to reach statistical significance. Antibiotic treatment during both the first and the second year of life increased the risk of AP in girls at 12 (OR 1.65; 95% CI: 1.09–2.49), but not in boys or the whole cohort. Conclusions & Inferences Antibiotic treatment does not seem to be a major risk factor for AP at 12 years. However, we cannot exclude that repeated courses, especially to infant girls, or use of broad-spectrum antibiotics between 9 and 12 years may be associated with an increased risk of AP.

Sense of Coherence (SOC) is hypothesized to have direct physiological consequences on endocrine and immunological processes. In this study, we compare parental SOC scores from pregnancy in groups of infants and parents representing different lifestyles (anthroposophic, partly anthroposophic and nonanthroposophic). We also analyse whether these could predict cortisol levels of the parents and their infants at 6-24 months postpartum.Parental SOC-13 was collected during the third trimester of pregnancy from a birth cohort of families with different lifestyles. Salivary samples were collected from the whole family when the child was 6 months (n = 210), 12 (n = 178) and 24 months of age (n = 149), and cortisol levels were analysed with radioimmunoassay technique.Sense of Coherence scores did not differ between the three lifestyle groups, and there were no correlations between SOC scores and salivary cortisol concentrations in separate analyses of mothers, fathers and children at any sampling age or at any sampling time during the day (morning, afternoon, bedtime).Sense of Coherence scores did not vary in parents with different lifestyles and were not associated with salivary cortisol levels in parents or in children.

To analyse salivary cortisol levels in 12- and 24-month-olds from families with an anthroposophic lifestyle and comparisons ('partly anthroposophic' and 'non-anthroposophic').Salivary samples were collected at child ages of 12 (n = 178) and 24 (n = 149) months. Cortisol was analysed with radioimmunoassay technique.Evening cortisol levels in children from anthroposophic families were lower than in comparisons at 12 months of age (geometric means: anthroposophic 1.7, partly anthroposophic 1.9, non-anthroposophic 3.6 nmol/L; p = 0.024) and at 24 months of age (1.1, 1.8 and 2.9 nmol/L, respectively; p = 0.002). At 24 months of age, similar differences were noted also for the afternoon levels (2.3, 3.3 and 3.9 nmol/L, respectively; p = 0.043). At age 12 months, the differences in the evening cortisol were statistically explained by a meat-free diet and at age 24 months by the anthroposophic lifestyle as such. The circadian variations were parallel in the three groups at age 12 and 24 months. No cortisol differences were observed between parents representing different lifestyles.An anthroposophic lifestyle is associated with low cortisol levels in the evening at age 12 and 24 months, at age 24 months also in the afternoon.

OBJECTIVE. Our aim was to investigate the role of measles vaccination and measles infection in the development of allergic disease and atopic sensitization. METHODS. A total of 14 893 children were included from the cross-sectional, multicenter Prevention of Allergy–Risk Factors for Sensitization in Children Related to Farming and Anthroposophic Lifestyle study, conducted in 5 European countries (Austria, Germany, the Netherlands, Sweden, and Switzerland). The children were between 5 and 13 years of age and represented farm children, Steiner-school children, and 2 reference groups. Children attending Steiner schools often have an anthroposophic (holistic) lifestyle in which some immunizations are avoided or postponed. Parental questionnaires provided information on exposure and lifestyle factors as well as symptoms and diagnoses in the children. A sample of the children was invited for additional tests, and 4049 children provided a blood sample for immunoglobulin E analyses. Only children with complete information on measles vaccination and infection were included in the analyses (84%). RESULTS. In the whole group of children, atopic sensitization was inversely associated with measles infection, and a similar tendency was seen for measles vaccination. To reduce risks of disease-related modification of exposure, children who reported symptoms of wheezing and/or eczema debuting during first year of life were excluded from some analyses. After this exclusion, inverse associations were observed between measles infection and “any allergic symptom” and “any diagnosis of allergy by a physician.” However, no associations were found between measles vaccination and allergic disease. CONCLUSION. Our data suggest that measles infection may protect against allergic disease in children.

Summary Background There is a close relation between the psychosocial environment of the infant—including the perception of maternal behaviour—and cortisol levels of the infant. One previous study has also demonstrated a correlation between mother and infant mean cortisol levels. In this study, this relation was further explored, also including father cortisol levels. Methods Saliva cortisol samples were collected from 51 six-months-olds and their parents on the same day in the morning, afternoon and evening. Analyses were performed with a radioimmunoassay technique. All mothers were at home with their child at this age and 47/51 mothers were breast feeding. Results Strong correlations were found between mother and child levels on all sampling occasions whereas weaker correlations were found between father and child levels and only in the afternoon and the evening samples. There was also a strong relation between waking up/bedtime-difference in mother and child and a weaker relation between the corresponding measure in father and child. Conclusions The stronger mother–infant than father–infant cortisol level correlations probably mirror that mother and infant not only have genetic similarities but also have been exposed to similar environmental conditions to a higher degree than father and infant.

Background The anthroposophic lifestyle has several features of interest in relation to allergy: for example, a restrictive use of antibiotics and certain vaccinations. In a previous Swedish study, Steiner school children (who often have an anthroposophic lifestyle) showed a reduced risk of atopy, but specific protective factors could not be identified. Objective To investigate factors that may contribute to the lower risk of allergy among Steiner school children. Methods Cross-sectional multicenter study including 6630 children age 5 to 13 years (4606 from Steiner schools and 2024 from reference schools) in 5 European countries. Results The prevalence of several studied outcomes was lower in Steiner school children than in the reference group. Overall, there were statistically significant reduced risks for rhinoconjunctivitis, atopic eczema, and atopic sensitization (allergen-specific IgE ≥0.35 kU/L), with some heterogeneity between the countries. Focusing on doctor-diagnosed disease, use of antibiotics during first year of life was associated with increased risks of rhinoconjunctivitis (odds ratio [OR], 1.97; 95% CI, 1.26-3.08), asthma (OR, 2.79; 95% CI, 2.03-3.83), and atopic eczema (OR, 1.63; 95% CI, 1.22-2.17). Early use of antipyretics was related to an increased risk of asthma (OR, 1.54; 95% CI, 1.11-2.13) and atopic eczema (OR, 1.32; 95% CI, 1.02-1.71). Children having received measles, mumps, and rubella vaccination showed an increased risk of rhinoconjunctivitis, whereas measles infection was associated with a lower risk of IgE-mediated eczema. Conclusion Certain features of the anthroposophic lifestyle, such as restrictive use of antibiotics and antipyretics, are associated with a reduced risk of allergic disease in children.