Pierantonio Macchia

Associations between Nasal Torquetenovirus Load and Spirometric Indices in Children with Asthma

Fifty-nine children with well-controlled, mild to moderate persistent asthma were studied for the presence and load of torquetenovirus (TTV) in nasal fluid. Rates of TTV positivity and mean nasal TTV loads were not dissimilar to those observed in the general population and in a group of 30 age- and residence-matched healthy control children without a history of asthmatic disease. However, in the children with asthma, 3 important indices of lung function--forced expiratory flow (FEF) in which 25% and 75% of forced vital capacity (FVC) is expired (FEF(25%-75%)), forced expiratory volume in 1 s/FVC, and FEF(25%-75%)/FVC--showed an inverse correlation with nasal TTV load. Furthermore, signs of reduced airflow were more frequent in the children with asthma who had high nasal TTV loads (> or =6 log(10) DNA copies/mL of nasal fluid) than they were in those who had low nasal TTV loads (<6 log(10) DNA copies/mL of nasal fluid), despite similar therapy regimens. In contrast, the control children showed no associations between nasal TTV load and the spirometric indices. Levels of eosinophil cationic protein in sputum were also greater in the children with asthma who had higher nasal viral burdens than they were in those who had lower nasal viral burdens. These findings are the first report of TTV infection status in children with asthma and suggest that TTV might be a contributing factor in the lung impairment caused by this condition.

Nasal nitric oxide in atypical primary ciliary dyskinesia.

BACKGROUND Atypical cases of primary ciliary dyskinesia (PCD) may present with minimal transmission electron microscopy (TEM) defects. The diagnostic role of nasal nitric oxide (nNO) levels was evaluated in those patients. METHODS Sixty-four children with recurrent pneumonia were studied with ciliary motion analysis, TEM, and nNO. RESULTS Investigations indicated PCD in 12 patients, secondary ciliary dyskinesia (SCD) in 50 patients, and normal results in 2 patients. In 4 of 50 children with SCD, atypical PCD was considered possible. The mean (+/- SD) nNO was 130 +/- 46.95 parts per billion in children affected by PCD, 127.79 +/- 68.58 parts per billion in atypical patients, and 760 +/- 221 parts per billion in children with SCD. Three to 5 months later, the nNO level was 132.75 +/- 55.76 parts per billion in children with atypical disease and 778.00 +/- 197 parts per billion in children with SCD. CONCLUSION Low levels of nNO may help to identify patients with atypical PCD.

Agenesis of paranasal sinuses and nasal nitric oxide in primary ciliary dyskinesia

Agenesis of paranasal sinuses has only been described in case reports of patients with primary ciliary dyskinesia (PCD). As agenesis of paranasal sinuses may contribute to low nasal nitric oxide levels, a common finding in PCD, we speculated that this condition might frequently occur in PCD patients. Patients referred for PCD evaluation were consecutively recruited for 30 months. In addition to standard diagnostic testing for PCD, a computed tomography (CT) scan of paranasal sinuses was performed in all subjects. 86 patients (46 children aged 8–17 yrs) were studied. PCD was diagnosed in 41 subjects and secondary ciliary dyskinesia (SCD) was diagnosed in the remaining 45 subjects. Frontal and/or sphenoidal sinuses were either aplastic or hypoplastic on CT scans in 30 (73%) out of 41 PCD patients, but in only 17 (38%) out of 45 with SCD (p = 0.002). There was a significant inverse correlation between the score for aplasia/hypoplasia of each paranasal sinus and nasal NO values in the PCD patients (p = 0.008, r = −0.432) but not in SCD (p = 0.07, r = −0.271). The findings of aplasia/hypoplasia of the frontal and or sphenoidal sinuses may be part of the spectrum of PCD and this finding should prompt exclusion of this condition.

Health-related quality of life and unmet needs in patients with primary ciliary dyskinesia

Few studies have evaluated the quality of life of patients with primary ciliary dyskinesia (PCD). We sought to determine the health impact of the disease as well as the unmet needs in a large group of patients. Questionnaires were either posted or e-mailed to known patients with PCD and published online. Questionnaires included the St Georges Respiratory Questionnaire, the Medical Outcomes Study Short Form-36 and a questionnaire that we produced to obtain information on age of diagnosis, symptoms and likely PCD-specific problems of these patients. 78 subjects (96% of those invited) answered all the questionnaires. Patients were diagnosed at a mean age of 9.4 yrs. Progressive worsening of the disease was observed and adherence to physiotherapy was found to be poor, particularly in adolescents and adults. Patients with the highest treatment burden had a worse quality of life. Over time patients become progressively less interested in treating their disease and adherence to treatment modalities decreases. PCD is associated with a progressive and continuous impact on the physical and mental health of the patients. Earlier identification of the patients and better strategies aimed at improving compliance with care are urgently needed.

Nasal nitric oxide and nitric oxide synthase expression in primary ciliary dyskinesia

No study has evaluated the correlation between different expression of nitric oxide synthase (NOS) isoforms in nasal epithelial cells and nasal NO (nNO) level in primary ciliary dyskinesia (PCD). Gene expression of endothelial (NOS3) and inducible NOS (NOS2) and their correlation with nNO level, ciliary function and morphology were studied in patients with PCD or secondary ciliary dyskinesia (SCD). NOS3 gene polymorphisms were studied in blood leukocytes. A total of 212 subjects were studied (48 with PCD, 161 with SCD and three normal subjects). nNO level correlated with mean ciliary beat frequency (p = 0.044; r = 0.174). The lower the nNO level the higher was the percentage of immotile cilia (p<0.001; r = -0.375). A significant positive correlation between NOS2 gene expression and nNO levels was demonstrated in all children (p = 0.001; r = 0.428), and this correlation was confirmed in patients with PCD (p = 0.019; r = 0.484). NOS2 gene expression was lower in PCD than in SCD (p = 0.04). The NOS3 isoform correlated with missing central microtubules (p = 0.048; r = 0.447). nNO levels were higher in PCD subjects with the NOS3 thymidine 894 mutation, and this was associated with a higher ciliary beat frequency (p = 0.045). These results demonstrate a relationship between nNO level, NOS mRNA expression and ciliary beat frequency.

New DNAH11 mutations in primary ciliary dyskinesia with normal axonemal ultrastructure.

To the Editors: In primary ciliary dyskinesia (PCD; Mendelian Inheritance in Man database #242650), a rare genetic disorder, the dysfunctional motility of cilia and impaired mucociliary clearance result in a myriad of clinical manifestations including recurrent infections of the respiratory tract, eventually causing lung damage, such as bronchiectasis, laterality defects and male infertility 1. The heterogeneous clinical presentation of PCD and the limitations of transmission electron microscopy (TEM) to assess ultrastructural defects within the cilium may delay the diagnosis 2. Ciliary beat frequency and pattern analyses and nasal nitric oxide (nNO) measurements are helpful 1, 3, 4, but only ciliogenesis in cultured cells has been reported as decisive for the diagnosis of the atypical PCD phenotype with normal axonemal ultrastructure 1, 5. Under these conditions, the identification of disease-causing mutations could overcome the current diagnostic limitations of TEM and improve our understanding of the biology and function of the cilium 2, 6, 7. For these reasons, we have assessed whether an analysis of the DNAH11 gene, in which some nonsense mutations have been reported to be associated with a normal axonemal ultrastructure but with an abnormal nonflexible beating pattern, reduced cilium bending capacity and a hyperkinetic beat 7, could be used to identify new mutations in three atypical PCD patients and thus be used in the diagnostic work-up of these most difficult cases. We observed three patients (A and B, sister and brother; and C, male), at the ages of 15 yrs and 5 months, 9 yrs and 4 months, and 8 yrs, respectively. The two siblings presented situs inversus. Both patients had chronic respiratory symptoms which required repeated treatment with antibiotics. Patient A also had neonatal purulent rhinitis with frequent relapses, chronic rhinosinusitis and otitis …

Widening of the clinical spectrum of Bartonella henselae infection as recognized through serodiagnostics

Abstract The recently improved diagnostics have widened, in children, the spectrum of clinical manifestations recognisable as Bartonella henselae infection. We report here the clinical features of 20 (14 males) consecutive children with serologically proved B. henselae infection observed within 12 months in the Paediatric Department of the University of Pisa. The patients had a mean age of 7 years 4 months (range 1.1–14.1 years). All children but one had a history of contact with kittens. Clinical manifestations included regional lymphadenopathy in 14 patients, representing in five the only clinical manifestation at onset, infectious mononucleosis-like syndrome in six, erythema nodosum in three, and Parinaud oculoglandular syndrome in one. In five patients a severe disorder was first suspected: fever of unknown origin in two with multiple hepatosplenic granulomatosis in one; osteolityc lesion suggesting bone neoplasm, marked inguinal lymph-node enlargement, suggesting Burkitt lymphoma, and an acute encephalopathy in one each. Bartonella henselae IgG antibody was positive in all patients with a titre ranging from 1:128 to 1:8590. IgM antibody was present in all except one child with an IgG titre of 1:2048. All patients recovered, some spontaneously. ConclusionBartonella henselae infection is frequent in Tuscany and probably underdiagnosed due to the high frequency of atypical onset of the clinical manifestations. An accurate clinical history and a reasonably wide use of the serological test may allow a rapid and accurate diagnosis, reassuring the family of the patient and avoiding invasive and expensive diagnostic procedures.

High torquetenovirus loads are correlated with bronchiectasis and peripheral airflow limitation in children.

Background: The aim of the study was to evaluate the prevalence of torquetenovirus (TTV) infection in a group of children with recurrent lower respiratory tract infections and radiologic evidence of bronchiectasis. Correlations between TTV loads and severity of bronchiectasis and between TTV loads and lung function were evaluated. Methods: In 38 subjects, high-resolution computed tomography (HRCT) and plasma tests for TTV detection and quantification were done. In 21/38 subjects, spirometry was also performed. Results: TTV was found in 31/38 (81.6%) patients. The correlation between TTV loads and severity of bronchiectasis was statistically significant (r = 0.548; P = 0.01). TTV loads showed inverse correlation with FEF25–75% (r = −0.541; P = 0.011), and FEF25–75%/FVC (r = −0.512; P = 0.018). Inverse correlation was found also between severity of bronchiectasis and functional lung parameters: FEF25–75% (r = −0.635; P = 0.002), FEV1/FVC (r = −0.541; P = 0.011), and FEF25–75%/FVC (r = −0.645; P = 0.002). Conclusions: This study demonstrated the high prevalence of TTV infection in children with bronchiectasis. Moreover, we have shown a significant correlation between TTV loads and airflow limitation within the peripheral airways, as well as between severity of bronchiectasis and decrease of lung function.

Simplified cell culture method for the diagnosis of atypical Primary Ciliary Dyskinesia

Background: The diagnosis of primary ciliary dyskinesia (PCD) can be challenging, and it may be particularly difficult to distinguish primary ciliary disease from the secondary changes after infections. Objectives: The purpose of the study was to evaluate if nasal epithelial cells, obtained with nasal brushing instead of a biopsy, could be used in a culture system for the diagnosis of PCD in difficult cases. Methods and main results: Ciliary motion analysis (CMA) and transmission electron microscopy (TEM) were performed on 59 subjects with persistent or recurrent pneumonia. These investigations allowed the diagnosis of PCD in 13 (22%) patients while the defect of the cilia was considered secondary to infections in 37 (63%) subjects. In the remaining nine (15%) patients the diagnostic evaluation with CMA and TEM remained inconclusive. Ciliogenesis in culture allowed the diagnosis of PCD in four of these patients, it was indicative of a secondary defect in two subjects, and it was not helpful in the remaining three patients. Conclusions: Culture of cells obtained with brushing of the nasal turbinate is not a perfect test, nevertheless it may offer diagnostic help in doubtful cases of PCD.

High Prevalence of Antibodies to Bartonella henselae among Italian Children without Evidence of Cat Scratch Disease

Few data are available on the seroprevalence of antibodies to Bartonella henselae among children. We retrospectively evaluated the presence of immunoglobulin G and M class antibodies to B. henselae in 508 children living in central Italy who were apparently free of any features suggesting B. henselae infection. We found that B. henselae infection is common among children in central Italy, occurs early in life, is in most cases asymptomatic, and resolves spontaneously.