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Background: Bronchial epithelial reticular basement membrane (RBM) thickening occurs in diseases with both eosinophilic (allergic bronchial asthma [BA]) and neutrophilic (cystic fibrosis [CF] and primary ciliary dyskinesia [PCD]) chronic airway inflammation; however, the lung function and airway remodeling relation remains unclear. The aim of this study was to test whether ventilation inhomogeneity is related to RBM thickening., Methods: Multiple breath washout test, endobronchial biopsy, and BAL were performed in 24 children with CF, 11 with PCD, 15 with BA, and in 19 control subjects. Lung clearance index at 2.5% (1/40th) of starting nitrogen concentration (LCI 2.5 ), RBM thickness, and lavage fluid cytology were quantified; their mutual associations were studied by using Spearman rank correlations (r)., Results: In asthma, ventilation inhomogeneity (mean ± SD) was mild (LCI 2.5 , 9.3 ± 1.4 vs 7.9 ± 0.9 in control subjects; P = .0391), and the RBM thickened (5.26 ± 0.98 μm vs 3.12 ± 0.62 μm in control subjects; P < .0001). No relation between RBM thickness and ventilation inhomogeneity or lavage cytology was found. In CF and PCD, RBM thickness was similar to that in asthma (4.54 ± 0.66 μm and 5.27 ± 1.11 μm, respectively), but ventilation inhomogeneity was significantly higher (LCI 2.5 , 12.5 ± 2.4 and 11.8 ± 2.5). Both in CF and PCD, RBM thickness correlated with LCI 2.5 (r = 0.594, P = .015; r = 0.821, P = .023). In PCD only, RBM thickness was also related to the number of neutrophils in lavage fluid (r = 0.821; P = .023)., Conclusions: Lung function impairment in relation to RBM thickness was milder in BA than in CF and PCD. In asthma, ventilation inhomogeneity did not correlate with RBM thickness, whereas it did in CF and PCD. This outcome suggests a different structure-function relation in these diseases., (Copyright © 2019 American College of Chest Physicians. Published by Elsevier Inc. All rights reserved.)

Background: Epithelial dysfunction has been implicated in asthma pathophysiology, but no studies have directly assessed ciliary function in asthma., Objective: To study the ciliary function and epithelial ultrastructure of patients with asthma and healthy controls., Methods: We studied ciliary beat frequency and beat pattern by using digital high-speed video imaging and ultrastructure by transmission electron microscopy of bronchial epithelial strips from 7 subjects with mild, 7 with moderate, and 19 with severe asthma and 9 healthy controls., Results: The median (interquartile range) ciliary beat frequency was decreased in moderate (6.5 [4.4-8.5] Hz) and severe asthma (6.7 [6.1-7.6] Hz) compared with controls (10.5 [9.7-11.8] Hz; P < .01). Dyskinesia and immotility indices were higher in severe asthma (65% [43%-75%]; 6.3% [1%-9.5%], respectively) compared with controls (4% [0%-6.7%; 0%, respectively; P < .01). These abnormalities were related to disease severity (ciliary beat frequency, r(s) = -0.68; dyskinesia index, r(s) = 0.86; immotility index, r(s) = 0.65; P < .0001). The ultrastructure of the epithelium was abnormal in severe asthma with a reduction in ciliated cells, an increase in dead cells, and ciliary disorientation compared with all other groups (P < .05). Compared with patients with mild asthma and healthy controls, patients with severe asthma showed increased ciliary depletion, microtubular defects, mitochondrial damage, and cytoplasmic blebbing (P < .01). All of these changes were related to disease severity., Conclusion: Ciliary dysfunction and ultrastructural abnormalities are closely related to asthma severity. Ciliary dysfunction is a feature of moderate to severe asthma, and profound ultrastructural abnormalities are restricted to severe disease. Whether these changes contribute to the development of severe asthma phenotype remains to be determined., (Copyright © 2010 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.)

Immotile cilia syndromes is a cause of recurrent infection of the airways and recurrent bronchopneumonias. Among the ciliary abnormalities are found changes in the structure of the microtubules, unco-ordinated ciliary movements caused by the absence of inner or outer or both dynein arms, and abnormalities of the kinetosomes and/or rete ridges. In patients with ciliary dyskinaesia bronchitis occurs early in life (during infancy) and usually has a recurrent tendency, so that bronchial biopsy is frequently undergone for diagnostic purposes. In this study we include 127 bronchial biopsies from patients (from 2 months to 49 years) unsuccessfully treated for recurrent respiratory tract infections. When performing regular diagnostic procedures on the light and electron microscopic level, we have looked for cilia abnormalities and also focused on changes within the mucosa and submucosa. The most common abnormality recorded was absence of the inner dynein arms, but in 40 cases neither of the dynein arms were present. Only a few patients had classical Kartagener's syndrome. Special attention is drawn to biopsies from elderly patients, in whom long-standing infections were followed by extensive damage to the bronchial epithelium, including even a total absence of ciliated cells. In some cases enhanced regenerative processes and some foci of squamous metaplasia were found. In two cases even foci of low-grade dysplasia were diagnosed.

Background: Ciliary ultrastructural defects with total lack of dynein arms (DA) cause abnormal mucociliary function leading to the chronic infections observed in primary ciliary dyskinesia. The role of partial ciliary ultrastructural defects, especially those involving the central complex, and their relationship with respiratory symptoms have been less thoroughly investigated., Objective: In a pediatric population with partial ciliary defects, we determined the relationship(s) between ultrastructural findings, ciliary motility, and clinical and functional features, and evaluated the outcome of this population., Design: We analyzed the clinical presentation and pulmonary function of 43 children with chronic bronchitis and partial ultrastructural defects (from 15% to 90%) of their respiratory cilia demonstrated on bronchial biopsies. The study population was divided into 3 groups according to ciliary ultrastructure: the main ultrastructural defect concerned the central complex in 23 patients (CC group), peripheral microtubules in 8 patients (PMT group), and DA in 12 patients (DA group)., Results: The percentage of ciliary defects was lower in the PMT group than in the CC and DA groups. Patients in the PMT group had less severe disease with frequent normal ciliary motility. Patients in the CC group had initially a higher incidence of respiratory tract infections, extensive bronchiectasis frequently requiring surgery, and arguments in favor of a congenital origin (high proportion of sibling form). Partial absence of DA, although of congenital origin, was associated with a good prognosis. In all groups, follow-up showed that the functional prognosis remained good with appropriate treatment., Conclusions: In children with chronic respiratory infections, presence of situs inversus, sibling form, obstructive pulmonary syndrome, or bronchiectasis required ultrastructural analysis, regardless of ciliary motility. Detection of CC abnormalities is a marker of severity and required intensive therapy and close follow-up.

Nasal and bronchial cilia and spermatozoa of a patient with a high clinical suspicion of a ciliary dyskinesia syndrome were ultrastructurally studied and quantified. Defective cilia showed two types of axonemal patterns: 9d+0s and 8d+1d. Of these, 9d+0s cilia prevailed in the proximal region, whereas 8d+1d prevailed in the distal region. Translocation of a peripheral doublet to the central position occurred at the middle region of cilia lacking the central pair, probably to compensate for its absence. Quantitative analysis showed that the percentages of anomalous cilia were 5.32+/-0.93 in nasal samples and 43.17+/-2.34 in bronchial samples. Spermatozoa without the central pair or with a translocated microtubular doublet were rarely observed, but a variety of nonspecific defects were seen. Even though transposition is generally considered to be an inherited ciliary defect and one of the causes of primary ciliary dyskinesia, in this case quantitative ultrastructural analysis and clinical data indicate that this is an acquired ciliary defect.

In the ciliary dyskinesia (immotile cilia syndrome) shows the partial or total lacking of cilia's elements. The ciliary dyskinesia may be developed congenital or acquired. The authors report on the experience with 72 biopsies from bronchial mucosa of 68 children, submitted with the question of immotile cilia syndrome. On micrographs (M: 64,000x) of the specimens processed by routine electron microscopical method the number of outer and inner dynein arm, A and B peripheral tubules, central tubules and central sheet were determined to normal 9 + 2 structure. 50-100 ciliaris per case were examined. Total or partial lacking of dynein and non-dynein elements were expressed for the total number of ciliaris compartments. Considering any earlier quantitative examination with this expression there was ease to characterised the quantitative behaviour of the components of ciliaris. Seven Kartagener's syndrome cases was the positive control for determined the quantitative differences between the primer and secondary ciliary changes. In the primary ciliary defects where the situs inversus were presented the total lacking of outer or/and inner dynein arms, where the situs inversus were not presented only the total lacking of inner dynein arms could be found. In secondary ciliary defects the partial lacking of the dynein arms and tubular components were presented. The used method is suitable to characterise the primary or secondary ciliary defects of bronchial mucosa.

The present study has the aim of studying the morphological aspects of ciliary abnormalities in patients affected by immotile-dyskinetik cilia syndrome. Five patients affected by primary ciliary dyskinesia have been studied, examining by TEM the cells obtained by bronchial brushing during fiberoptic bronchoscopy. Many types of anomalies have been observed, and a morphologic classification was proposed. The frequency of abnormal cilia appears to be correlated to the severity of respiratory pathology. The morphological anomalies are not specific for this pathology.

Objective: The objective of this study was to investigate whether reduced bronchial mucus transport velocity (BTV) is associated with a loss of cilia or ultrastructural abnormalities of cilia in intubated patients., Design: The patients were studied prospectively in a convenience sample trial., Setting: The study took place in a university hospital., Patients and Participants: 29 orally intubated patients in a surgical ICU., Interventions: BTV was measured with radiolabeled microspheres in the right and left primary bronchus. Following these measurements, biopsy samples were taken from the bronchi for scanning (SEM) and transmission (TEM) electron-microscopic investigations., Measurements and Results: SEM: Patients with normal or slight impaired BTV (group 1, n = 14: BTV: 8.5 mm/min (3.8-11.5); median with range) showed more cilia on the luminal surface than patients with markedly depressed BTV (p < 0.05) (group 2, n = 15: BTV: 0 (0-2.1)). The difference was statistically significant. The BTV values correlated moderately with the number of cilia on the luminal surface (r = 0.46; p = 0.02). TEM: In group 1, 6.5% (3.9-14.9) of cilia were abnormal (median with range) vs 9.3% (4.9-13.7) in group 2; these differences were not statistically significant. Neither was there any significant correlation between BTV and the frequency of abnormal cilia., Conclusions: Impaired mucociliary transport in intubated patients is associated with a loss of cilia rather than ultrastructural abnormalities of cilia, which are less relevant.

Ciliary dyskinesia is characterized by recurrent respiratory tract infections secondary to abnormal ciliary structure and function. The diagnosis of ciliary dyskinesia is based on the detection of axonemal ultrastructural abnormalities (AUA) is respiratory mucosa samples. In most cases, the diagnosis of AUA is made on samples obtained from nasal ciliated cells with little discomfort to the patient. However, no studies have been performed in the same patient to confirm whether nasal samples reflect bronchial ciliary changes. To answer this question and to determine whether it is necessary to sample bronchial cells for the diagnosis of ciliary dyskinesia, we investigated 12 patients (between the age of 5 and 63 yr) with chronic sputum production. The presence of situs inversus, bronchiectasis, chronic sinusitis, and sterility was investigated to determine an inherited disorder. Two groups were established: Group 1 = six patients with an inherited disorder and Group 2 = six patients without evidence of an inherited disorder. Samples were obtained by brushing or biopsy of nasal and bronchial mucosa and were processed for transmission electron microscopy. In Group 1, the mean AUA was 65.2 +/- 11.4%. The following predominant axonemal defects were found: absence of dynein arms (DA) (four patients), central complex abnormalities (CC) (one patient), and various AUA (one patient). Nasal and bronchial samples correlated significantly for total AUA (r' = 1, p < 0.01) and for outer DA defects (r' = 0.96, p < 0.05). A good but not significant correlation was found for inner DA (r' = 0.83) and peripheral microtubule (PM) defects (r' = 0.71). In Group 2, the mean AUA was 9.6 +/- 2.3%.(ABSTRACT TRUNCATED AT 250 WORDS)

The Authors during an extensive applications of nasal brushing in infancy according to Rutland and Cole, remarked--first time in literature--the contemporary presence in a child of the nemaline myopathy syndrome and immotile cilia syndrome. Supposing the possibility that in other myopathy may be present the same ciliary immotile syndrome, they studied with nasal brushing three cases of Werdnig-Hoffman syndromes: but they presented normal ciliary conformation. The Authors suppose that in case in future it should be found in other cases of nemaline myopathy the contemporary presence of dinein arms lack, it should be demonstrated that this syndrome takes derivation from a genetic alteration both of the muscular apparatus and the ciliary mobility system of bronchial epithelius.

The examination of 98 patients with newly diagnosed destructive pulmonary tuberculosis was undertaken to find the relation between the condition of the bronchial tree, level of obstruction and degree of mucociliary insufficiency. The latter was shown to depend on the condition of the bronchial free and was most manifest in diffuse endobronchitis. At the same time, the degree and level of obstruction of the bronchial tree also had influence on the rate of mucociliary transport.

Ciliary orientation was studied in 43 patients with the "immotile cilia" syndrome. Twenty-four of these patients had total situs inversus. One mucosal specimen was taken from uterine cervical epithelium, 2 were from bronchial mucosa and 40 from nasal mucosa. The orientation of the cilia was measured from micrographs using a semiautomatic image analyzer (IBAS I). The results from patients were compared with those of 10 control subjects. The mean standard deviation and its standard deviation of the angles of ciliary orientation was 39.7 degrees +/- 9.2 degrees in 43 patients and 27.4 degrees +/- 4.3 degrees in the control group. The difference between the groups is highly significant statistically (P less than or equal to 0.001). However, there were no statistically significant differences in the standard deviations of ciliary orientation between the fields sectioned near the cell membrane or near the ciliary tip. We were also unable to find any significant differences in the standard deviations of the ciliary angles in the specimens taken from brush biopsies and excisional biopsies. There were also no statistically significant differences between the standard deviations of the ciliary angles for the groups with or without situs inversus. If 35 degrees is considered to be the limit value for the mean standard deviation between normal and pathological specimens in our total material, this would give a specificity of 0.90 and a sensitivity of 0.72.

Recurrent bronchopulmonary and E.N.T. infections in a 2-month old child with complete situs inversus suggested an immotile cilia syndrome. Electron microscopy of the respiratory epithelium cilia demonstrated an ultra structural abnormality (defective radial spokes) typical of this recently discovered syndrome. This case is similar to 3 other cases in infants reported in the literature. It shows that the clinical manifestations of ciliary dysfunction may occur soon after birth and that early detection is desirable for optimal treatment. The type of abnormality detected and the percentage of cilia affected demonstrate that the syndrome is congenital and not acquired and provide information of the degree of ciliary dyskinesia. The genetic aspects are discussed. A diagnosis of "immotile cilia" syndrome should systematically be envisaged in infants with recurrent pneumonia or otitis of unknown aetiology, or when the clinical context (situs inversus, family history) is suggestive of the conditions.

The apical structure of human respiratory cilia was studied using thin and thick sectioning techniques. The distal end of the mature cilium is narrowed and presents dense material and a crown-shaped glycocalyx. The crown is composed of three to seven bristles, having a mean length of 31 nm and showing a periodic structure. The capping structure of the apical part of human respiratory cilia is composed of a plate and an electron-dense cap tightly bound to the central and to each of the nine peripheral A microtubules. Lateral spokes link the peripheral microtubules to the internal ciliary plasma membrane and dense filaments radiate from the dense cap to the ciliary crown. These ciliary apical structures may be involved in several functions associated with microtubule assembly as well as ciliary movement and mucociliary transport.

Ultrastructurally atypical bronchial cilia are studied and semiquantitatively analysed in 24 children suffering from recurrent respiratory tract infections with or without bronchiectasis. In patients with Kartagener's syndrome normal-looking and shortened dynein arms are present at some axonemal microtubular doublets. This finding suggests that the polymerization or assemblage of dynein molecules on microtubules only is defective but not totally lacking. Bilateral, local and partial absence of dynein arms is demonstrated in some of the patients with acquired unilateral bronchiectases. These patients also reveal anomalies of the "9 + 2" microtubular axonemal pattern. It is suggested that these abnormalities of the tubulin-dynein system are local and acquired defects that may impair bronchial mucociliary clearance. None of the patients with pneumonia and asthma or with cystic fibrosis studied show any anomalies of the dynein arms. However aberrant axonemal microtubular patterns and other ciliopathies such as naked axonemes and megacilia are present at times in these patients. We postulate that these atypical cilia are secondary acquired abnormalities. Only some patients with bacterial or viral pneumonia demonstrate a partial lack of dynein arms in bronchial cilia. Other ciliopathies such as megacilia, naked and intracytoplasmic axonemes and apical blebs are more frequent and more common in these patients. We suppose they manifest a secondary and rather aspecific pathogenic influence upon the bronchial ciliary substructure.

Congenital ciliary aplasia was demonstrated in two siblings with clinical history of primary ciliary dyskinesia. Ultrastructural histochemistry of successive bronchial biopsies revealed the predominance of immature mucous cells and the total absence of ciliated or preciliated cells in the respiratory epithelium. This original disorder may represent a unique variant of primary ciliary dyskinesia with primitive disregulation of ciliogenesis.

The fine structure of the basal apparatus of cilia from respiratory epithelium has been studied in 5 subjects with immotile cilia syndrome without any specific defects in ciliary morphology. The ultrastructural analysis revealed the presence of abnormalities, heretofore undescribed, in basal bodies and accessory structures. In particular defective basal bodies of the so-called "half centriole" type, abnormal basal bodies showing an unusual dense granule in their central part, as well as abnormal accessory structures (i.e., giant striated roots, and double basal foot arising from single basal bodies) were found to be variously distributed in the patients examined. The significance of these morphological changes is discussed.

Twenty-two cases displaying potentially precancerous epithelial changes of the bronchial mucosa--including basal and goblet cell hyperplasia, epidermoid metaplasia, and dysplasia--showed a wide variety of cilial abnormalities on the ultrastructural level. The changes comprised abnormal configurations of the ciliary plasma membrane, variations in the amount of ciliary matrix, disorganization of basal bodies, and diverse alterations of the axonemal microtubular pattern. More than one such alteration was observed in one and the same case, and there was no correlation with the actual type of epithelial lesion nor the degree of dysplasia. These changes, interpreted as abnormal ciliary regeneration in severely damaged ciliated cells, are potentially reversible and seem to represent a nonspecific response of the affected cells to an ample spectrum of noxes. Their diagnostic significance is poor, especially with regard to the modification of the axonemal microtubular pattern, unless the entire clinical picture suggests an immotile cilia syndrome.

Respiratory cilia of the bronchial mucosa from a 5-year-old boy with clinical evidence of classical Kartagener's syndrome (situs inversus, bronchiectasis and sinusitis) were first examined by means of transmission electron microscopy for identification of the axonemal defects described as typical for primary ciliary dyskinesia (PCD). Additional oscillography was performed on the cilia in vitro, which showed absence of a coordinated ciliary beat frequency. After clear classification of the case as PCD, a freeze-fracture examination of the respiratory cilia was performed, which revealed a higher density of intramembrane particles on the outer fracture face (E-face) than on the inner fracture face (P-face). The results were discussed with regard to probable pathogenetic aspects on PCD.

Human respiratory cilia, including those in seven cases of immotile cilia syndrome, were examined by transmission electron microscopy after tannic acid-containing fixation. This fixation method is useful for ultrastructural observation of unit membranes and of the dynein arms and protofilaments of cilia. In addition, intramembrane particles and electron-dense strands were also demonstrated in the neck of the cilia. These structures may correspond to the ciliary necklace, which has been demonstrated by the freeze-etch technique. In high-magnification electron micrographs, the intramembrane particles were 10 nm in diameter and the necklace spacing was about 30 nm in human respiratory cilia. Prior to this study, no reports on ultrastructural abnormalities of the ciliary necklace in humans had appeared, probably because of technical problems with tissue preparation. Therefore tannic acid-containing fixation appears to be useful for ultrastructural observation of the ciliary necklace.