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1.  Peanut Sensitization Profiles in Italian Children and Adolescents with Specific IgE to Peanuts 
BioMed Research International  2013;2013:170452.
Peanuts are one of the most relevant foods implicated in IgE-mediated adverse reactions in pediatric population. This study aimed to evaluate the pattern of sensitization towards five peanut allergenic components (rAra h 1, 2, 3, 8 and 9) in a population of Italian children and adolescents with specific IgE (sIgE) to peanut. rAra h 9 was the main allergen implicated in peanut sensitization (58%), followed by rAra h 8 (35%), rAra h 2 (27%), rAra h 3 (23%) and rAra h 1 (12.5%). rAra h 1, 2, and 3 were the main allergenic components in young children: 8/13 (62%) between 2 and 5 years, 8/23 (35%) between 6 and 11 years, and 3/12 (25%) between 1 and 16 years. No differences were found among the levels of sIgE towards rAra h 1, 2, 3, and 9 in the three groups; in contrast, the levels of sIgE against rAra h 8 showed an increasing trend according to age. In conclusion rAra h 1, 2, and 3 were the prevalent sensitizing allergens during the first years of life in Italian patients with sIgE to peanuts (“genuine” allergy); in contrast rAra h 9 and 8 were mainly involved in school-age children and adolescents with pollen allergy (“secondary” sensitization).
doi:10.1155/2013/170452
PMCID: PMC3845422  PMID: 24324955
2.  Skin prick test to foods in childhood atopic eczema: pros and cons 
Skin prick tests are the first investigation in allergy diagnostics and their use is described in all the guidelines on atopic eczema. However, the clinical usefulness of skin prick tests is the subject of great debate. On the one hand, skin prick tests allow the identification both of individuals at risk for food allergy and of the allergen inducing the eczematous flare. On the other hand, when performed by a non-specific specialist, positive skin prick tests to foods may wrongly lead to prolonged elimination diets, which may induce nutritional deficiencies and perhaps loss of tolerance to the avoided foods. Furthermore, skin prick tests increase health costs. A consensus on this topic has not yet been reached. Considering the diversity of clinical stages in which it occurs, atopic eczema presentation should be the starting point to determine whether or not skin prick tests should be carried out.
doi:10.1186/1824-7288-39-48
PMCID: PMC3734168  PMID: 23902622
Atopic dermatitis; Atopic eczema; Skin prick test; Food allergy
3.  A qRT-PCR assay for the expression of all Mal d 1 isoallergen genes 
BMC Plant Biology  2013;13:51.
Background
A considerable number of individuals suffer from oral allergy syndrome (OAS) to apple, resulting in the avoidance of apple consumption. Apple cultivars differ greatly in their allergenic properties, but knowledge of the causes for such differences is incomplete. Mal d 1 is considered the major apple allergen. For Mal d 1, a wide range of isoallergens and variants exist, and they are encoded by a large gene family. To identify the specific proteins/genes that are potentially involved in the allergy, we developed a PCR assay to monitor the expression of each individual Mal d 1 gene. Gene-specific primer pairs were designed for the exploitation of sequence differences among Mal d 1 genes. The specificity of these primers was validated using both in silico and in vitro techniques. Subsequently, this assay was applied to the peel and flesh of fruits from the two cultivars ‘Florina’ and ‘Gala’.
Results
We successfully developed gene-specific primer pairs for each of the 31 Mal d 1 genes and incorporated them into a qRT-PCR assay. The results from the application of the assay showed that 11 genes were not expressed in fruit. In addition, differential expression was observed among the Mal d 1 genes that were expressed in the fruit. Moreover, the expression levels were tissue and cultivar dependent.
Conclusion
The assay developed in this study facilitated the first characterisation of the expression levels of all known Mal d 1 genes in a gene-specific manner. Using this assay on different fruit tissues and cultivars, we obtained knowledge concerning gene relevance in allergenicity. This study provides new perspectives for research on both plant breeding and immunotherapy.
doi:10.1186/1471-2229-13-51
PMCID: PMC3616815  PMID: 23522122
Apple allergy; OAS; Mal d 1; Bet v 1; PR-10; gene family; qRT-PCR
4.  Citrus Allergy from Pollen to Clinical Symptoms 
PLoS ONE  2013;8(1):e53680.
Allergy to citrus fruits is often associated with pollinosis and sensitization to other plants due to a phenomenon of cross-reactivity. The aims of the present study were to highlight the cross-reactivity among citrus and the major allergenic pollens/fruits, throughout clinical and molecular investigations, and to evaluate the sensitization frequency to citrus fruits in a population of children and adults with pollinosis. We found a relevant percentage of sensitisation (39%) to citrus fruits in the patients recruited and in all of them the IgE-mediated mechanism has been confirmed by the positive response to the prick-to-prick test. RT-PCR experiments showed the expression of Cit s 1, Cit s 3 and a profilin isoform, already described in apple, also in Citrus clementine pollen. Data of multiple sequence alignments demonstrated that Citrus allergens shared high percentage identity values with other clinically relevant species (i.e. Triticum aestivum, Malus domestica), confirming the possible cross-allergenicity citrus/grasses and citrus/apple. Finally, a novelty of the present work has been the expression of two phospholipaseA2 isoforms (PLA2 α and β) in Citrus as well as in Triticum pollens; being PLA2 able to generate pro-inflammatory factors, this enzyme could participate in the activation of the allergenic inflammatory cascade.
doi:10.1371/journal.pone.0053680
PMCID: PMC3537725  PMID: 23308273
5.  Unbalance of intestinal microbiota in atopic children 
BMC Microbiology  2012;12:95.
Background
Playing a strategic role in the host immune function, the intestinal microbiota has been recently hypothesized to be involved in the etiology of atopy. In order to investigate the gastrointestinal microbial ecology of atopic disease, here we performed a pilot comparative molecular analysis of the faecal microbiota in atopic children and healthy controls.
Results
Nineteen atopic children and 12 healthy controls aged 4–14 years were enrolled. Stools were collected and the faecal microbiota was characterized by means of the already developed phylogenetic microarray platform, HTF-Microbi.Array, and quantitative PCR. The intestinal microbiota of atopic children showed a significant depletion in members of the Clostridium cluster IV, Faecalibacterium prausnitzii, Akkermansia muciniphila and a corresponding increase of the relative abundance of Enterobacteriaceae.
Conclusion
Depleted in key immunomodulatory symbionts, the atopy-associated microbiota can represent an inflammogenic microbial consortium which can contribute to the severity of the disease. Our data open the way to the therapeutic manipulation of the intestinal microbiota in the treatment of atopy by means of pharmaceutical probiotics.
doi:10.1186/1471-2180-12-95
PMCID: PMC3404014  PMID: 22672413
6.  End point prick test: could this new test be used to predict the outcome of oral food challenge in children with cow's milk allergy? 
Background
Cow's milk allergy (CMA) is the most frequent food allergy in childhood; the trend of CMA is often characterized by a progressive improvement to achieve tolerance in the first 4 to 5 years of life.
It has been observed that specific IgE (sIgE) towards cow's milk proteins decrease when the age increases.
Although food allergy can be easily diagnosed, it is difficult to predict the outcome of the oral food challenge (OFC), that remains the gold standard in the diagnosis of food allergy, by allergometric tests.
Methods
We considered 44 children with CMA diagnosed through OFC who returned to our Allergy and Immunology Pediatric Department between January to December 2010 to evaluate the persistence of allergy or the achievement of tolerance.
On the basis of the history, we performed both allergometric skin tests and OFC in children that were still following a milk-free diet, whereas only allergometric skin tests those that had already undergone spontaneous introduction of milk protein at home without presenting symptoms.
Objective
The aim of this study was to investigate the relationship between the persistence of CMA or the acquisition of tolerance and the results of the end point prick test (EPT).
Results and Discussion
The OFC with cow's milk was performed on 30 children, 4 children were excluded because of a history of severe reactions to cow's milk, and 10 because they had spontaneously already taken milk food derivates at home without problems. 16/30 (53%) children showed clinical reactions and the challenge was stopped, 14/30 (47%) did not have any reaction.
Comparing the mean wheal diameter of every EPT's dilution between the group of allergic children and the tolerant ones, we obtained a significant difference (p < 0.05) for the first 4 dilutions.
We have also calculated sensitivity (SE), specificity (SP), the positive predictive value (PPV) and the negative predictive value (NPV) for each EPT dilution.
Conclusions
EPT is a safe and cheap test, easy to be executed and that could provide good prediction of the outcome of OFC; so it might be used to avoid OFC-induced anaphylaxis in children affected by CMA. It can also help avoiding dietetic restrictions in tolerant children who show sensitization towards cow's milk proteins.
doi:10.1186/1824-7288-37-52
PMCID: PMC3220633  PMID: 22053846
Cow's milk proteins allergy; end point prick test; food oral challenge; tolerance
7.  Atopic Dermatitis and the Atopic March: What Is New? 
Journal of Allergy  2011;2011:279425.
Objective. In this paper the authors review the management of atopic dermatitis (AD) and the association between AD and allergic respiratory diseases. Data Sources. PubMed databases, researching articles in the last 15 years. Results. Studies about atopic march are cross-sectional population studies at different ages. They show that the most important predisposing factor for atopy is a decrease of the filaggrin's expression. Conclusions. The most modern theories seem to show that the most important factor which starts the atopic march is represented by an impaired epidermal barrier. It causes an increase in skin permeability to allergens that could induce sensitization even in the airways. The major predisposing factor is a primary inherited epithelial barrier defect resulting from filaggrin gene mutation, but other factors may play a role in this complex mechanism. Further studies are needed to focus on AD treatment and preventive strategies.
doi:10.1155/2011/279425
PMCID: PMC3173727  PMID: 21941575
9.  Pooled Genome-Wide Analysis to Identify Novel Risk Loci for Pediatric Allergic Asthma 
PLoS ONE  2011;6(2):e16912.
Background
Genome-wide association studies of pooled DNA samples were shown to be a valuable tool to identify candidate SNPs associated to a phenotype. No such study was up to now applied to childhood allergic asthma, even if the very high complexity of asthma genetics is an appropriate field to explore the potential of pooled GWAS approach.
Methodology/Principal Findings
We performed a pooled GWAS and individual genotyping in 269 children with allergic respiratory diseases comparing allergic children with and without asthma. We used a modular approach to identify the most significant loci associated with asthma by combining silhouette statistics and physical distance method with cluster-adapted thresholding. We found 97% concordance between pooled GWAS and individual genotyping, with 36 out of 37 top-scoring SNPs significant at individual genotyping level. The most significant SNP is located inside the coding sequence of C5, an already identified asthma susceptibility gene, while the other loci regulate functions that are relevant to bronchial physiopathology, as immune- or inflammation-mediated mechanisms and airway smooth muscle contraction. Integration with gene expression data showed that almost half of the putative susceptibility genes are differentially expressed in experimental asthma mouse models.
Conclusion/Significance
Combined silhouette statistics and cluster-adapted physical distance threshold analysis of pooled GWAS data is an efficient method to identify candidate SNP associated to asthma development in an allergic pediatric population.
doi:10.1371/journal.pone.0016912
PMCID: PMC3040188  PMID: 21359210
10.  Does improvement management of atopic dermatitis influence the appearance of respiratory allergic diseases? A follow-up study 
Background
Atopic dermatitis (AD) is often the prelude to allergic diseases. The aim of this study was 1) to evaluate if an integrated management regime could bring about a change in the evolution of the disease in comparison to the results of a previous study; 2) to determine whether the refinement of allergic investigations allowed to identify more promptly the risk factors of evolution into respiratory allergic diseases.
Methods
The study included 176 children affected by AD and previously evaluated between 1993 and 2002 at the age of 9-16 months, who underwent a telephonic interview by means of a semi-structured, pre-formed questionnaire after a mean follow-up time of 8 years. According to the SCORAD, at first evaluation children had mild AD in 23% of cases, moderate in 62%, severe in 15%.
Results
AD disappeared in 92 cases (52%), asthma appeared in 30 (17%) and rhinoconjunctivitis in 48 (27%). The factors significantly related to the appearance of asthma were: sensitization to food allergens with sIgE > 2 KU/L (cow's milk and hen's egg; P < 0.05); to inhalant allergens with sIgE > 0.35 KU/L (P < 0.05). Logistic regression analysis showed that inhalant sensitization was positively related to the occurrence of asthma (OR = 4.219). While AD showed similar rates of disappearance to those of our previous study, the incidence of asthma was reduced, at the same follow-up time, from 29% to 15% (P = 0.002), and the incidence of rhinoconjunctivitis from 35% to 24% (P = 0.02).
Conclusion
Comparing the results with those of the previous study, integrated management of AD does not seem to influence its natural course. Nevertheless, the decrease in the percentage of children evolving towards respiratory allergic disease stresses the importance of early diagnosis and improvement management carried out by specialist centers. The presence of allergic sensitization at one year of age might predict the development of respiratory allergy.
doi:10.1186/1476-7961-8-8
PMCID: PMC2907296  PMID: 20591145
11.  Use of the Italian version of the Pediatric Asthma Quality of Life Questionnaire in the daily practice: results of a prospective study 
BMC Pediatrics  2009;9:30.
Background
Asthma is a serious global health problem and its prevalence is increasing, especially among children. It represents a significant social and economic burden, and it can severely affect the health-related quality of life (HRQL) of patients. Among the numerous questionnaires aiming at evaluating asthma HRQL in children, the Pediatric Asthma Quality of Life Questionnaire (PAQLQ) has proved to have good measurement properties.
The present study was aimed at investigating the possible role of the Italian, self-administered version of the PAQLQ in the routine clinical evaluation of children affected by bronchial asthma.
Methods
52 Italian children and adolescents (40 males and 12 females), aged 6 to 17 years, affected by allergic asthma, were enrolled. Each patient was evaluated twice, and at each visit asthma control and severity were assessed, spirometry was performed and the patients completed the self-administered version of the PAQLQ.
Results
The questionnaire was well-accepted and understood by the children. Children showed an overall good quality of life, with mild impairment in the activity and emotional function domains. The PAQLQ showed an overall good correlation with the clinical and functional indexes that are normally evaluated in follow-up visits of asthmatic patients. The PAQLQ appeared to be strongly related to asthma control, both at the first (p < 0.01) and second (p < 0.001) time of the study. The PAQLQ was also seen to decrease with increasing asthma severity. The results suggest a better compliance of the children towards completion of the questionnaire at t1. Finally, the PAQLQ does not appear to discriminate HRQL in patients with good lung function.
Conclusion
The Italian version of the PAQLQ is a quick-to-administer aid to clinical activity and can add valuable information to symptom reports, objective measurements and clinical assessment of asthma control and severity in daily clinical practice. Re-administration at each follow-up visit allows HRQL to be monitored over time.
doi:10.1186/1471-2431-9-30
PMCID: PMC2685371  PMID: 19422700

Results 1-11 (11)