Risk factors for recurrent acute otitis media: a real-life clinical experience.

Acute otitis media (AOM) is the most common bacterial infection in children. Some children with AOM tend to be otitis-prone, such as frequent recurrence of AOM (RAOM). Possible RAOM risk factors are widely debated. The current study was performed in a real-life setting, such as an otorhinolaryngologic (ORL) clinic, to identify predictive factors, including clinical data and endoscopic findings, for RAOM in children. In this study, 1,002 children (550 males, 452 females, mean age 5.77 + 1.84 years) complaining of upper airway symptoms were consecutively visited. Detailed clinical history and nasal endoscopy were performed. Throughout the ORL visit, it was possible to define some factors involved in the recurrence of AOM, including female gender, artificial feeding, tonsillar and adenoid hypertrophy. Adenoid and tonsillar hypertrophy, female gender, and artificial are factors significantly associated with RAOM. Therefore, reducing adenoid and tonsil size, also using topical corticosteroids or glycyrrhizin, could be a reasonable strategy to potentially reduce adenoid and tonsil size. The current study suggests that also in a primary care setting, it is possible to achieve meaningful information that is relevant in clinical practice.

Can an otorhinolaryngological visit induce the suspect of allergic rhinitis in children?

Summary: Allergic rhinitis (AR) is very frequent in childhood. AR is commonly associated with some co-morbidities and typical clinical features. This study aimed to test the hypothesis whether an otorhinolaryngological (ORL) visit could induce the suspect of AR. Globally, 1,002 children (550 males, mean age 5.77 years) were consecutively visited at an ORL clinic. Clinical visit, nasal endoscopy, and skin prick test were performed in all patients. In particular, history investigated atopic familiarity, birth, feeding type, passive smoking, comorbidities, including asthma, respiratory infections, otitis media, respiratory sleep disorder. Endoscopy assessed the tonsil and adenoid volume, turbinate contacts, mucosal color, and nasal discharge. Univariate and multivariate analysis were performed. The study showed that 547 (54.6%) children had AR. Some parameters were predicting factor for suspecting AR: middle turbinate contact (OR = 9.27), familial atopy (OR = 6.24), pale nasal mucosa (OR = 4.95), large adenoid volume (OR = 3.02 for score 4), and asthma co-morbidity (OR = 2.95). In conclusion this real-life study showed that during an ORL visit it is possible to suspect AR in children with turbinate hypertrophy, familial atopy, nasal pale mucosa, adenoid enlargement, and asthma comorbidity.

Bet v 1 sensitization modulates allergenic molecular immune response.

Summary: Background Allergy is characterized by allergen-specific IgE production. Molecular-based allergy diagnostic allows to define the precise sensitization profile. Bet v 1 is the major allergen of the PR-10 family. It has been reported that pan-allergens could affect the sensitization panel in adults. This study aimed to evaluate the impact of Bet v 1 sensitization on sensitization pattern in a large sample of children. Methods Serum IgE molecular components were assessed by ISAC method. Sera from 1,205 children, 708 males (58.76%) and 497 females (41.24%), median age 8.61 years (4.93 - 12.54 years) were analyzed. Results A total of 354 PR-10-positive subjects were detected out of 1,205 subjects. Bet v 1 positive children were significantly more frequently sensitized to other molecules belonging to PR-10 family and noteworthy also to other allergenic families than Bet v 1 negative children. Conclusions The present study demonstrates that Bet v 1 sensitization may significantly affect the sensitization pattern in children living in Genoa, a Mediterranean city located in a birch-free area.

Diagnostic relevance of IgE sensitization profiles to eight recombinant Phleum pratense molecules.

BACKGROUND: Grass pollen-related seasonal allergic rhinoconjunctivitis (SARg) is clinically heterogeneous in severity, comorbidities, and response to treatment. The component-resolved diagnostics disclosed also a high heterogeneity at molecular level. Our study aimed at analyzing the characteristics of the IgE sensitization to Phleum pratense molecules and investigating the diagnostic relevance of such molecules in childhood. METHODS: We examined 1120 children (age 4-18 years) with SARg. Standardized questionnaires on atopy were acquired through informatics platform (AllergyCARD™). Skin prick tests were performed with pollen extracts. Serum IgE to airborne allergens and eight P. pratense molecules (rPhl p 1, rPhl p 2, rPhl p 4, rPhl p 5b, rPhl p 6, rPhl p 7, rPhl p 11, rPhl p 12) were tested by ImmunoCAP FEIA. RESULTS: The analysis of IgE responses against eight P. pratense molecules showed 87 profiles. According to the number of molecules recognized by IgE, the more complex profiles were characterized by higher serum total IgE, higher grass-specific serum IgE, and higher number and degree of sensitization to pollens. The most frequent IgE sensitization profile was the monomolecular Phl p 1. Sensitization to Phl p 7 was a reliable biomarker of asthma, whereas Phl p 12 of oral allergy syndrome. Sensitization to Phl p 7 was associated with a higher severity of SARg, and complex profiles were associated with longer disease duration. CONCLUSIONS: In a large pediatric population, the complexity of IgE sensitization profiles against P. pratense molecules is related to high atopic features although useless for predicting the clinical severity. The detection of serum IgE to Phl p 1, Phl p 7, and Phl p 12 can be used as clinical biomarkers of SARg and comorbidities. Further studies in different areas are required to test the impact of different IgE molecular profiles on AIT response.

The impact of age on serum allergen-specific IgE to inhaled molecular components.

BACKGROUND: Respiratory allergy is characterised by an IgE-mediated reaction. The immune system functions, including IgE production, progressively decline over time, such as growing up and ageing. Molecular-based allergy diagnostic defines sensitisation profile. This study aimed to evaluate the impact of age on serum allergen-specific IgE to molecular component levels in a large sample of subjects. METHODS: Serum IgE to: Phl p1, Bet v1, Ole e1, Cup a1, Par j2, Can f1, Der p2, and Fel d1 were assessed by ISAC method. Sera from 2788 patients, 1230 males (44.1%) and 1558 females (55.9%), median age 23 years (1st and 3rd quartiles: 9.7-49.7 years; age range: 1 month-103 years) were analysed. RESULTS: The number of positive tests (i.e. sensitisation) tended to increase between birth and school-age till young adulthood and then decreased (p<0.0001) with the exception of Fel d 1 (p=0.14). A similar age-dependent trend was observed considering the levels of each allergen components: the levels of each allergen component, with the exception of Fel d 1, tended to increase till early adulthood and then to decrease reaching the lowest levels in the elderly. CONCLUSIONS: Allergen-specific IgE production to inhaled molecular components trend to reduce with ageing, but with differences between allergens. This phenomenon should be adequately evaluated managing allergic patients.

Allergen-specific IgE to food molecular components and age: From early childhood to adulthood.

BACKGROUND: Respiratory allergy is characterised by an IgE-mediated reaction. The immune system functions, including IgE production, progressively decline over time with growing up and ageing. Molecular-based allergy diagnostic defines sensitisation profile. This study aimed to evaluate the impact of age on serum allergen-specific IgE to molecular component levels in a large sample of subjects. METHODS: Serum IgE to: rCor a11, rPru p3, nJug r3, rAra h8, rGly m4, rCor a8, nPen m1, nAct d8, Bos d 8, and nGal d2 were assessed by ISAC method. Sera from 2795 patients, 1234 males (44.1%) and 1561 females (55.9%), median age 23 years (1st and 3rd quartiles: 9.7-43.7 years; age range: 1 month-103 years) were analysed. RESULTS: The number of positive tests (i.e. sensitisation) tended to increase between birth and school-age until young adulthood and then decreased. A similar age-dependent trend was observed considering the levels of each allergen components: the levels of each allergen component tended to increase until early adulthood, but Gal d 2 and Bos d 8 (rapidly diminishing), and then to decrease over time. However, the pattern is significantly dependent on each single tested food. CONCLUSIONS: Allergen-specific IgE production to food molecular components tend to reduce with ageing, but with differences between allergens. This phenomenon should be adequately evaluated managing allergic patients.

Asthma exacerbation in children: relationship among pollens, weather, and air pollution.

BACKGROUND: Asthma exacerbation is an episode of (sub)acute worsening of asthmatic symptoms. Exacerbation may depend on environmental factors. OBJECTIVE: The present study investigated emergency calls for asthma exacerbation in children, analysing: i) their trend over the course of time; and ii) their possible relationship with environmental factors, including pollen count, meteorological parameters, and air pollution. METHODS: Emergency calls for exacerbation were recorded for 10 years (from 2002 to 2011) in Genoa (Italy). Betulaceae, Urticaceae, Gramineae, and Oleaceae pollen counts were measured. Meteorological parameters and air pollutants were also measured in the same area and for the same period. RESULTS: The number of emergency calls did not significantly modify during the time studied. Two main peaks were detected: during the autumn and the spring. Wind speed significantly diminished as did most air pollutants. There were significant and relevant relationships between emergency calls and: pollens during the spring (r = 0.498), rainfall (r = 0.818), wind speed (r = 0.727), and air pollutants (r = 0.622 for SO2; r = 0.699 for NO; r = 0.58 for NO2). CONCLUSIONS: This 10-year survey demonstrates that: (i) asthma exacerbations did not diminish over the time; (ii) there were seasonal peaks (autumn and spring); (iii) pollens (mainly Parietaria), wind speed and rainfall, SO2, NO, O3 and NO2 were strongly associated with asthma exacerbations in children in this area. Therefore, asthma exacerbations may significantly depend on environmental variations.

Nasal high-mobility group box-1 protein in children with allergic rhinitis.

BACKGROUND: Allergic rhinitis (AR) is characterized by an inflammatory reaction. High-mobility group box-1 protein (HMGB1) has many characteristics similar to classic proinflammatory cytokines. No study has yet investigated its role in AR. The aim of this study was to measure HMGB1 levels in the fluid recovered from nasal lavage in children with untreated AR and in control subjects. MATERIALS: The study was conducted on 104 AR subjects (48 males and 56 females, median age 10.3 ± 3.4 years) and 97 healthy children (42 males and 55 females) who were age-matched (median age 9.8 ± 4.1 years). Total serum immunoglobulin E, peripheral eosinophils and nasal symptoms assessed by visual analog scale (VAS) were considered. HMGB1 was measured using an ELISA assay. RESULTS: HMGB1 levels in nasal lavage fluid were higher in AR children than in the control group (96.9 ± 19.3 vs. 9.27 ± 4.01 ng/ml; p < 0.001). There was a very strong relationship between HMGB1 levels and VAS values in AR children (r = 0.919). Considering the symptom severity assessed by VAS, there was a relationship between HMGB1 and VAS in all AR subgroups: more evident in the severe subgroup (r = 0.727). CONCLUSIONS: Nasal HMGB1 has significantly increased in children with AR and is significantly related to symptom severity.

Formulation and characterization of silk fibroin films as a scaffold for adipose-derived stem cells in skin tissue engineering.

Skin substitutes are epidermal, dermal or complete bilayered constructs, composed by natural or synthetic scaffolds and by adherent cells such as fibroblasts, keratinocytes or mesenchymal stem cells. Silk fibroin is a promising polymer to realize scaffolds, since it is biocompatible, biodegradable, and exhibits excellent mechanical properties in terms of tensile strength. Moreover, fibroin can be added of others components in order to modify the biomaterial properties for the purpose. The aim of this work is to prepare silk fibroin films for adipose-derived stem cell (ADSCs) culture as a novel feeder layer for skin tissue engineering. Pectin has been added to promote the protein conformational transition and construct strength, while glycerol as plasticizer, providing biomaterial flexibility. Eighteen formulations were prepared by casting method using fibroin, pectin (range 1-10% w/w), and glycerol (range 0-20% w/w); films were characterized by Fourier transform infrared spectroscopy and differential scanning calorimetry assay, to select the optimal composition. A stable fibroin conformation was obtained using 6% w/w pectin, and the best mechanical properties were obtained using 12% w/w glycerol. Films were sterilized, and human ADSCs were seeded and cultured for 15 days. Cells adhere to the support assuming a fibroblastic-like shape and reaching confluence. The ultrastructural analysis evidences typical active-cell features and adhesion structures that promote cell anchorage to the film, thus developing a multilayered cell structure. This construct could be advantageously employed in cutaneous wound healing or where the use of ADSCs scaffold is indicated either in human or veterinary field.

Clinical presentation and pathogenesis of cold-induced autoinflammatory disease in a family with recurrence of an NLRP12 mutation.

OBJECTIVE: NLRP12 mutations have been described in patients affected with peculiar autoinflammatory symptoms. This study was undertaken to characterize NLRP12 mutations in patients with autoinflammatory syndromes, particularly a novel missense mutation, p.D294E, affecting a protein sequence crucial for ATP binding, which was identified in a Caucasian family with familial cold-induced autoinflammatory syndrome in some family members. METHODS: Fifty patients were tested for NLRP12 mutations. A Caucasian family with the p.D294E missense mutation of NLRP12 in some family members was clinically characterized. In vitro analysis of the effects of the mutation on NF-κB activity was performed in HEK 293 cells after cotransfection of the cells with a luciferase NF-κB-responsive element and mutant or wild-type (WT) NLRP12 expression plasmids. NF-κB activity was also evaluated 24 hours after stimulation with tumor necrosis factor α in monocytes from individual family members carrying the mutation. Furthermore, secretion of interleukin-1ß (IL-1ß), production of reactive oxygen species (ROS), and activation of antioxidant systems in patient and healthy donor monocytes, under resting conditions and after stimulation with pathogen-associated molecular patterns (PAMPs), were also assessed. RESULTS: In the family assessed, the p.D294E mutation segregated in association with a particular sensitivity to cold exposure (especially arthralgias and myalgia), but not always with an inflammatory phenotype (e.g., urticarial rash or fever). In vitro, the mutant protein maintained the same inhibitory activity as that shown by WT NLRP12. Consistently, NLRP12-mutated monocytes showed neither increased levels of p65-induced NF-κB activity nor higher secretion of IL-1ß. However, the kinetics of PAMP-induced IL-1ß secretion were significantly accelerated, and high production of ROS and up-regulation of antioxidant systems were demonstrated. CONCLUSION: Even with a variable range of associated manifestations, the extreme sensitivity to cold represents the main clinical hallmark in an individual carrying the p.D294E mutation of the NLRP12 gene. Although regulation of NF-κB activity is not affected in patients, redox alterations and accelerated secretion of IL-1ß are associated with this mild autoinflammatory phenotype.

Impaired spirometry may suggest sensitization.

The present study confirms that sensitization is very frequent in the general population and suggests that impaired FEF25-75 may be a marker of sensitization. Therefore, when spirometry is abnormal, mainly concerning FEF25-75, sensitization should be suspected.

Impaired FEF25-75 may predict high exhaled nitric oxide values in children with allergic rhinitis and/or asthma.

Allergic rhinitis and asthma are closely associated. Inflammation is a common pathological characteristic shared by both disorders. The measure of the fractional concentration of exhaled nitric oxide (FeNO) may be considered a surrogate marker for airway inflammation. Forced expiratory flow between 25 and 75 percent of vital capacity (FEF25-75) has been previously demonstrated to be able to predict BHR and bronchial reversibility. The aim of this study was to evaluate whether impaired FEF25-75 values may be related to FeNO values in a pediatric cohort of allergic subjects. 850 children with allergic rhinitis, allergic asthma, or both, were evaluated. Bronchial function (FEV1, FVC, and FEF25-75), FeNO, and sensitizations were assessed. Bronchial function and FeNO were significantly different in the 3 groups (p less than 0.001). A strong inverse correlation between FeNO and FEV1was found in patients with rhinitis, asthma and asthma with rhinitis (r= -0.72, r=-0.70 and r= -0.70, respectively). Impaired FEF25-75 values (such as less than 65 percent of predicted) were significantly associated with high FeNO levels (such as =34 ppb). In conclusion, this study provided evidence that FEF25-75 is strongly and inversely related with FeNO and FEF25-75 may predict high FeNO levels in children with allergic rhinitis, asthma or both.

Impaired FEF25-75 values may predict bronchial reversibility in allergic children with rhinitis or asthma.

FEV1 is considered an important parameter for asthma diagnosis and follow-up. However, it has been proposed that FEF25-75 could be more sensitive than FEV1 to detect slight airways obstruction. Bronchial reversibility defined by positive response to bronchodilation test. The aim of the present study was to define whether an impaired FEF25-75 value (less than 65 percent of predicted) may be predictive for reversibility in a large cohort of allergic children with rhinitis or asthma. Six hundred allergic children were recruited: 300 with controlled asthma and 300 with allergic rhinitis. All of them were evaluated by performing spirometry, bronchodilation test, and skin prick test. Two predictors were significantly associated with bronchial reversibility: i) an impaired FEF25-75 value (less than 65 percent of predicted), and ii) sensitization to perennial allergens. It was more relevant in children with rhinitis (ORAdj:8.9 and 2.2 respectively). In conclusion, this study, conducted in real life, could suggest that an impaired FEF25-75 value (less than 65 percent of predicted) may be considered a reliable marker of bronchial reversibility, mainly in children with allergic rhinitis.

Serum IL-23 in asthmatic children.

Asthma is characterized by airway inflammation that is controlled by a complex cytokine network. The Th1/Th2 imbalance has been well documented in the pathogenesis of allergic asthma. Recently, Th17 cells and regulatory T (Treg) cells have been found to participate in the pathogenesis of allergic asthma. This study aimed at verifying whether anti-inflammatory treatment could change serum IL-4, IL-10 and IL-23 in asthmatic children. Globally, 78 children (40 males and 38 females, median age 9.3 +- 3.7 years), with asthma and monosensitized to house dust mites, were evaluated. Lung function (such as FEV1) and serum IL-4, IL-10 and IL-23 levels were measured at baseline (T0), after 4 weeks (T1) and after 12 weeks (T2) of inhaled corticosteroid (ICS) treatment. The control group consisted of 40 healthy children (22 males and 18 females) age matched. At baseline, IL-4 and IL-23 levels were higher in severe asthmatics than in control group (p less than 0.001), while serum IL-10 levels were significantly lower in group of asthmatic children as compared to healthy control group (p less than 0.001). At T2, IL-4 and IL-23 significantly diminished (p less than 0.001), while IL-10 significantly increased. There was significant relationship between FEV1 and IL-4, IL-10 and IL-23 at T0 (r=-0.784; r=-0.735 and r=-0.787, respectively). Moreover, there were correlations between FEV1 and IL-4, IL-10 and IL-23 in patients at T1 (r=-0.563; r=-0.539 and r=-0.583, respectively) and at T2 (r=-0.549; r=-0.428 and r=-0.393, respectively). The present study provided evidence that: i) serum IL-23 was up-regulated also in asthmatic children, ii) ICS treatment was able of reducing IL-23, and iii) IL-23 change well related with lung function improvement. Thus, it is presumable that IL-23 could be a suitable marker of allergic inflammation in asthma.

Bronchodilation test in patients with allergic rhinitis.

BACKGROUND: Allergic rhinitis (AR) may be considered a risk factor for the onset of asthma. Recently, it has been reported that forced expiratory flow between 25% and 75% of vital capacity (FEF25₋75) may predict a positive response to bronchodilation test in asthmatic children. The aim of this study was to evaluate a large group of adult AR patients to investigate the frequency of response to bronchodilation test and FEF25₋75 values. METHODS: One thousand four hundred and sixty-nine consecutive patients suffering from persistent AR were evaluated. Clinical examination, spirometry, and bronchodilation test were performed in all patients. RESULTS: In this study, 62.9% of patients had reversibility to bronchodilation test and 17.8% had impaired FEF25₋75 values (≤ 65% of predicted). Impaired FEF25₋75 values associated with longer rhinitis duration may predict reversibility to bronchodilation test (OR = 11.3; P < 0.001). In addition, a FEF25₋75 cutoff value ≤ 71% of predicted may already discriminate patients with reversibility. CONCLUSIONS: This study highlights that about two-thirds of patients with persistent AR may be considered at risk of becoming asthmatic. This finding should be adequately considered as a precocious spirometry may allow the early detection of patients prone to develop asthma and consequently to treat them.