Simplified cell culture method for the diagnosis of atypical primary ciliary dyskinesia

Pifferi, Massimo; Montemurro, Francesca; Cangiotti, Angela Maria; Ragazzo, Vincenzo; Cicco, Maria Di; Vinci, B; Vozzi, Giovanni; Macchia, Pierantonio; Boner, Attilio

doi:10.1136/thx.2008.110940

Cited by 41 publications

(31 citation statements)

References 28 publications

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“…This may relate to different techniques or equipment, and it is important for each centre to establish their own normal ranges [19]. The diagnosis of PCD was confirmed by tissue culture in all patients with higher than expected nNO levels [16], and so we are confident that the diagnosis is correct. Secondly, it was only possible to study NOS2 and NOS3 mRNA, not protein, so the possibility cannot be excluded that post-translational modification may have affected the results.…”

Section: Discussionmentioning

confidence: 99%

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

Pifferi¹,

Bush²,

Maggi³

et al. 2011

European Respiratory Journal

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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 (p50.044; r50.174). The lower the nNO level the higher was the percentage of immotile cilia (p,0.001; r5 -0.375). A significant positive correlation between NOS2 gene expression and nNO levels was demonstrated in all children (p50.001; r50.428), and this correlation was confirmed in patients with PCD (p50.019; r50.484). NOS2 gene expression was lower in PCD than in SCD (p50.04). The NOS3 isoform correlated with missing central microtubules (p50.048; r50.447). nNO levels were higher in PCD subjects with the NOS3 thymidine 894 mutation, and this was associated with a higher ciliary beat frequency (p50.045).These results demonstrate a relationship between nNO level, NOS mRNA expression and ciliary beat frequency.

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Section: Discussionmentioning

confidence: 99%

“…In all subjects, ciliary motion analysis, ultrastructural assessment of cilia on nasal brushings and nNO measurement were performed. nNO measurements, nasal brush biopsy, ciliary motion analysis and ultrastructural studies were performed according to standard methodologies [12][13][14][15][16], detailed in the online supplementary material.…”

Section: Subjectsmentioning

confidence: 99%

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

Pifferi¹,

Bush²,

Maggi³

et al. 2011

European Respiratory Journal

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“…A diagnosis of PCD was made on the basis of structural and/or functional ciliary abnormalities, as described previously [6][7][8][9][10]. In all subjects, ciliary motion analysis, ultrastructural assessment of cilia, ciliary function after ciliogenesis in culture of nasal brushing samples, and nasal NO measurements were performed when the subject had been free from respiratory infection for o4 weeks.…”

Section: Methodsmentioning

confidence: 99%

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

Pifferi¹,

Bush²,

Caramella³

et al. 2010

European Respiratory Journal

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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 (p50.002). There was a significant inverse correlation between the score for aplasia/hypoplasia of each paranasal sinus and nasal NO values in the PCD patients (p50.008, r5 -0.432) but not in SCD (p50.07, r5 -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.

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“…Definite diagnosis of PCD was made on the basis of structural and/or functional ciliary abnormalities on both brushing and culture, which take account of both beat frequency and pattern, as previously described [7]. Diagnosis of SCD was made when PCD was excluded and ciliary abnormalities were considered as acquired.…”

Section: Subjectsmentioning

confidence: 99%

“…In the past, culture was mainly performed on cells obtained by nasal forceps biopsy [6], which is painful. Recently, we have demonstrated that cell culture is possible directly from the brushed nasal epithelial cells, thus reducing the need for a second and more traumatic sampling [7]. With our simplified culture system, the evaluation of ciliary activity is only possible on the 21st day of suspension culture, when new cilia formation has definitely occurred.…”

mentioning

confidence: 99%

Rapid diagnosis of primary ciliary dyskinesia: cell culture and soft computing analysis

et al. 2012

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Diagnosis of primary ciliary dyskinesia (PCD) sometimes requires repeated nasal brushing to exclude secondary ciliary alterations. Our aim was to evaluate whether the use of a new method of nasal epithelial cell culture can speed PCD diagnosis in doubtful cases and to identify which are the most informative parameters by means of a multilayer artificial neural network (ANN).A cross-sectional study was performed in patients with suspected PCD. All patients underwent nasal brushing for ciliary motion analysis, ultrastructural assessment and evaluation of ciliary function after ciliogenesis in culture by ANN.151 subjects were studied. A diagnostic suspension cell culture was obtained in 117 nasal brushings. A diagnosis of PCD was made in 36 subjects (29 of whom were children). In nine out of the 36 patients the diagnosis was made only after a second brushing, because of equivocal results of both tests at first examination. In each of these subjects diagnosis of PCD was confirmed by cell culture results.Cell culture in suspension evaluated by means of ANN allows the separation of PCD from secondary ciliary dyskinesia patients after only 5 days of culture and allows diagnosis to be reached in doubtful cases, thus avoiding the necessity of a second sample.

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Simplified cell culture method for the diagnosis of atypical primary ciliary dyskinesia

Abstract: Background:

Cited by 41 publications

References 28 publications

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

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

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

Rapid diagnosis of primary ciliary dyskinesia: cell culture and soft computing analysis

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