Ultrastructural changes of the nasal mucosa in primary ciliary dyskinesia

Primary ciliary dyskinesia syndrome (PCD) is a rare, autosomal receive disorder. Kartagener's syndrome is a subgroup of the PCD with situs inversus, bronchiectasis, and sinusitis. The symptoms results from an abnormal ultrastructural morphology of the cilia such as absence of dynein arms and other changes. As a consequence ciliary motility is disturbed. A 25-year-old man was examined because he suffered from recurrent severe pneumonia and Aspergillus infections of the lungs. On electron micrographs, ciliary abnormalities including deficiency of inner and outer dynein arms, dysmorphic outer dynein arms, and disorientation of the cilia were demonstrated. The diagnosis of PCD requires electron-microscopic investigations of the ciliated mucosa. Special attention should be given to ultrastructural changes of nasal or bronchial mucosa if a young patient suffers from recurrent severe respiratory infections.     

Ultrastructural study of immotile cilia syndrome

Recently immotile cilia syndrome has gained the interest of a number of investigators from the aspects of the physiology and pathology of the ciliary movement. This is because microstructural abnormalities of the dynein arms in the cilia of the respiratory mucosal epithelium and in the flagella of sperm tails have been identified in this syndrome. The present study was designed to find a simple clinical method for detecting patients having this syndrome, and was conducted to elucidate the clinical significance and etiology of this syndrome. In order to detect patients with immotile cilia syndrome, 72 patients with one of more conditions such as sinusitis, bronchiectasis, situs inversus and sterility were examined using a ciliary function test and electron microscope observation of the nasal cilia. Seven of the examined patients were diagnosed as having immotile cilia syndrome on the basis of the presence of the characteristic ultrastructural patterns of the nasal cilia, i.e., disorders of the dynein arms. The possibility of positive test results increases greatly as the degree of the complications increases, especially in the case of a combination of chronic inflammation of the respiratory tract with situs inversus and sterility. As the electron microscope findings of this syndrome, Afzelius (1979) has reported defects of the dynein arms, spoke head and central sheath. Beyond these microstructural abnormalities, I have identified abnormal attachment of the dynein arms as a new parameter in the electron microscopic diagnosis of immotile cilia syndrome.     

Abnormalities of cilia and chronic sinusitis

An ultrastructural study was performed on 34 biopsy samples of the sinusal mucosa in 28 patients who were investigated for chronic sinusitis. Twelve specimens with normoplastic sinusitis and eighteen with hyperplastic sinusitis were studied. Four normal specimens served as controls. The incidence of ultrastructural ciliary abnormalities was 2%. Morphological changes of dynein arms have not been observed in the present study. Compound cilia were found in approximately 2/3 of the studied cases. Microtubular abnormalities occurred in roughly 50%. The observed abnormalities, also seen in the control group, were independent on the sinusitis type and could not been correlated with mucociliary transport. Their significance is discussed.     

Ciliary ultrastructure in a child with Kartagener's syndrome

Summary Kartagener's syndrome has been characterized by a primary ultrastructural abnormality of the cilia which consequently impairs their movements. We used transmission electron microscopy with tannic acid staining to investigate the fine structure of the cilia from the nasal mucosa of a 7-year-old girl with Kartagener's syndrome. The staining technique employed was useful for visualizing the dynein arms and protofilaments of the microtubules of the cilia. Although 15% of the cilia examined demonstrated microtubular disarrangements, these findings were considered to be acquired changes due to chronic sinusitis. No abnormal ciliary ultrastructures specific to Kartagener's syndrome, such as absence of dynein arms, were detected in this study. In such cases without any abnormal ciliary ultrastructures, it is conceivable that some other unknown factor may be involved in the impaired ciliary movement.     

Ultrastructural expression of primary ciliary dyskinesia after ciliogenesis in culture

During the period 1990-1999 84 PCD patients were identified and characterized. The expression of inherited abnormalities in primary ciliary dyskinesia after ciliogenesis was investigated in 41 patients with dynein deficiency, 6 patients with absence of the central pair of microtubules and 24 PCD patients with normal ultrastructure. In patients with dynein deficiency, the outer dynein arms counts were 1.9 +/- 1.0 in the biopsies and 1.6 +/- 0.7 after ciliogenesis. Secondary abnormalities were found in 15.8 +/- 20.4% of the transverse sections of cilia and only in 1.0 +/- 1.3% after ciliogenesis. Ciliary orientation was 28 +/- 11 degrees and 24 +/- 10 degrees respectively in biopsies and cultures. In patients with absence of the central pair this was found in 15 +/- 16% in biopsies and 21 +/- 19% after ciliogenesis. The values for the outer dynein arm were 8.4 +/- 0.3 and 8.7 +/- 0.2 and for the secondary abnormalities were 11.7 +/- 7.3% and 0.5 +/- 1.3% in the biopsies, respectively after ciliogenesis. In patients with normal ultrastructure the scores for the dynein arms were similar. Secondary abnormalities were found in 12.2 +/- 11.7% in the biopsies and 0.6 +/- 0.9% after ciliogenesis while ciliary orientation was respectively 21 +/- 7 degrees and 25 +/- 8 degrees. In conclusion, inherited abnormalities in primary ciliary dyskinesia are expressed after ciliogenesis, while secondary abnormalities are virtually absent, thereby facilitating the ultrastructural diagnosis.     

Dynein arms and spokes after ciliogenesis in cultured respiratory epithelial cells from non-PCD individuals

Dynein arms and spokes are crucial components of cilia. Reference values for the dynein arms and spokes were calculated based on biopsies from non-PCD patients as well as after ciliogenesis in culture. The mean values in the biopsies (n = 251) were 8.4 +/- 0.5, 2.9 +/- 0.7 and 4.7 +/- 1.0 for the outer dynein arms, inner dynein arms and spokes respectively. After ciliogenesis in culture (n = 462) identical values were found: 8.7 +/- 0.4, 3.0 +/- 0.4 and 5.5 +/- 0.6. The lower limits of normality can be set at 7.0 and 1.2 for the outer and inner dynein arms respectively. The dynein arms and spokes were not influenced by the percentage of secondary abnormalities. In conclusion, dynein arms and spokes are readily identified after ciliogenesis in culture. These parameters are independent of secondary ciliary dyskinesia.     

Ultrastructural abnormalities of the cilia in human nasal epithelia

Nasal mucosal cilia were observed with electron microscope in 14 patients with immotile cilia syndrome (ICS), 9 with nasal papilloma (NP), 23 with sinobronchial syndrome (SB), 2 with sinusitis combined with dextrocardia (SC), 1 with Kartagener's syndrome (KS), and 5 normal controls (C). Abnormalities such as complex cilia, cilia with abnormal axonemes and cilia with randomly oriented central microtubules were frequently found in the groups of ICS (8.1%) and NP (10.4%) while less in other groups: SB (4.9%), SC (5.3%), KS (4.7%) and C (3.9%). The percentage of cilia with defective dynein arms (DA) was the highest in the ICS group (94.0%), followed by the groups of SC (53.7%), SB (47.5%), NP (41.2%), C (35.8%) and KS (33.3%). The ICS group was found to be the largest in the number of defective DA per a cilium (4.1), followed by the groups of NP (1.0), SB (0.6), SC (0.7), KS (0.4) and C (0.4). Increased rates of defective DA were also recognized in cilia of tracheal mucosa and flagella of sperm in 7 patients with ICS examined. In conclusion, neither abnormal cilia nor defective DA of cilia are specific findings for ICS. However, when we observe these findings in high percentage in nasal mucosa as well as in other organs, we may define this condition as ICS.     

A pediatric case of primary ciliary dyskinesia caused by novel copy number variation in PIH1D3

An 11-month-old boy with productive cough was referred to our hospital. He had nasal obstruction immediately after birth, and wheezing, wet cough, and rhinorrhea were observed daily after the neonatal period. Clinical and imaging findings revealed secretory otitis media, chronic sinusitis, and bronchiectasis. Primary ciliary dyskinesia was suspected. Transmission electron microscopy of nasal cilia showed defects of the outer and inner dynein arms. Genetic examinations of the family revealed copy number variation in PIH1 domain-containing 3 (PIH1D3) in the proband and mother. This is the first report of a Japanese patient with primary ciliary dyskinesia caused by copy number variation in PIH1D3.     

The immotile cilia syndrome — One cause of persistent upper respiratory tract infection

Four patients with persistent recurrent upper respiratory tract infections are presented. Electron microscopic studies of respiratory mucosal biopsies from these patients reveal ultrastructural abnormalities of cilia consisting of partial to complete loss of dynein arms, radial spoke disruption, and compound cilia. The concept of immotile cilia as an important cause of recurrent infections is receiving more attention. This study stresses the need for simple, rapid screening tests for cilial activity followed by electron microscopic evaluation in selected cases.     

Otitis media and the immotile cilia syndrome

The immotile cilia syndrome appears to be a congenital defect in the ultrastructure of cilia that renders them incapable of movement. Respiratory tract cilia and sperm are predominantly affected. Bronchiectasis, sinusitis and male sterility are the main clinical findings. Situs inversus may be found. To these findings can be added otitis media. The defect appears to be a complete or partial absence of dynein arms which are believed to be essential for generating movement of cilia or sperm tails. Six patients suspected of having immotile cilia were compared to six patients in a control group. In affected patients, no cilia movement in the middle ear or nasopharynx was observed using the operating microscope. Electron microscopy of cilia from the mucosa of the middle ear and nasopharynx appeared to confirm the ultrastructural defect in two of six patients suspected of having the syndrome.     

Characterization of a chemotactic defect in patients with Kartagener syndrome

Kartagener syndrome (KS) is an autosomally inherited recessive condition characterized by situs inversus, bronchiectasis, and chronic sinusitis. Ciliary dynein, the mechanochemical force generator in ciliary movement, is deficient in patients with KS. We examined blood samples from two patients and tissue biopsy specimens from five patients and found: (1) no significant defect in neutrophil or monocyte chemotaxis in response to formylpeptide chemoattractant; (2) no alterations in centriolar structure, but significantly more centriole-associated microtubules in KS neutrophils and monocytes than in control leukocytes; and (3) a marked reduction in KS fibroblast chemotaxis in response to fibronectin compared with control fibroblasts. The significance of these cellular defects in KS is described.     

Long-term follow-up of the clinical relevance of short outer dynein arms in human nasal cilia

Clinical significance of short outer dynein arms was examined in a long-term follow-up study of 76 patients with various respiratory symptoms. Clinical evaluations, nasal mucociliary transport rate (NMTR) measurement and transmission electron microscopy were performed. Follow-up examinations took place 5–11 years later. In the initial examination four patients and on follow-up seven patients were found to have short outer dynein arms in their nasal mucosal biopsies. Short dynein arms were associated with a slow NMTR, poorly coordinated ciliary beat direction, and clinical symptoms of perennial rhinitis and recurrent sinusitis. Short dynein arms were found to be a significant histologic finding and probably represent a variant of primary ciliary dyskinesia. In some cases short outer dynein arms may be a reversible finding with improving clinical symptoms. It is obvious that at least part of dynein defects are acquired. Received: 23 December 1997 / Accepted: 15 May 1998     

Unusual ciliary abnormalities in three 9/11 response workers

After the 9/11 terrorist attacks on the World Trade Center in New York in 2001, thousands of response workers were exposed to complex mixtures of toxins, pollutants, and carcinogens. Many developed illnesses involving the respiratory tract. We report unusual ultrastructural ciliary abnormalities in 3 response workers that corresponded to their respiratory and ciliary functional abnormalities. Each patient had respiratory cilia biopsies that were evaluated for motility and ultrastructural changes. Impaired ciliary motility was seen in 2 of the 3 patients. Each of the patients showed monomorphic ultrastructural abnormalities. Two of the patients showed identical triangular disarray of axonemal microtubules with peripheral doublets 1,4, and 7 forming the corners of the triangle and doublet 9 always more medially displaced than doublets 2, 3, 5, 6, and 8. Two workers had cilia in which axonemes were replaced by homogeneously dense cores. One of these also had cilia with triangular axonemes as previously described. The other had cilia with a geometric triangular to pentagonal shape. The ciliary abnormalities described here may represent a new class of primary ciliary dyskinesia in which abnormalities may have a genetic basis and a phenotypic expression that is prompted at the cellular level by local environmental conditions.     

Ciliary ultrastructure in patients with chronic rhinosinusitis and primary ciliary dyskinesia

The Cilia represent one of the main mechanisms contributing to the clearance of microorganisms and particles from the respiratory epithelium. Primary ciliary dyskinesia (PCD) is a genetically determined disorder characterized by irreversible systemic dysmotility of the cilia. Secondary ciliary dyskinesia (SCD) differs from primary defects on the reversible ultrastructural alterations that can occur after any insult to a previously normal mucosa. Hence, this study aimed to describe and compare the main ultrastructural ciliary features in PCD and SCD through transmission electron microscopy. The most frequent PCD abnormalities were missing or short dynein arms, missing central microtubules, and displacement of one of the nine peripheral doublets. The most common changes found in SCD were compound cilia and peripheral microtubule alterations associated with modifications of the respiratory epithelium. PCD presented a higher percentage of altered cilia (>30 %) when compared to SCD (5 %), demonstrating that SCD is more limited in area than PCD. Whereas in PCD the changes in the dynein arms and in the central microtubules are fundamental for diagnostic confirmation, the diagnosis of SCD usually involves compound cilia and disarrangements in peripheral microtubules.     

Changes in nasal epithelium in patients with severe chronic sinusitis: A clinicopathologic and electron microscopic study

Objective: Defective ciliary ultrastructure and impaired mucociliary clearance play an important role in the development of respiratory disease and sinusitis. Changes in the ciliary ultrastructure of the sinonasal epithelium have been documented in patients with primary ciliary dyskinesia. However, secondary ciliary dyskinesias and epithelial cytopathologic changes have been underappreciated as a consequence of respiratory dysfunction and chronic sinusitis. Study Design: Thirty-two patients with severe chronic sinusitis were evaluated for ciliary and epithelial abnormalities. Materials and Methods: Fourteen patients (44%) were children who underwent full allergy, sweat, and immunologic workups. Eighteen patients (56%) were adults who had severe refractory sinusitis and had failed previous sinus surgery. All patients underwent nasal epithelium biopsies of the middle turbinate and evaluation by light and transmission electron microscopy. Results: Ciliated cells were found in 23 patients (72%) with 9 patients (28%) having no cilia. Foci of normal ciliated epithelium were found in only 19% of the patients, often in epithelial invaginations. Variable numbers (usually a minor population) of cilia in 20 cases (87%) exhibited ultrastructural defects including compound cilia and microtubule and dynein arm defects. All of the patients showed variable loss of differentiated epithelial cells ranging from denuded epithelium to basal cell hyperplasia often associated with squamous metaplasia, secondary to chronic sinonasal disease. The lamina propria was often edematous with dilated capillaries, plasma cells, lymphocytes, and hyperplastic seromucous glands. Conclusions: This study demonstrates that ciliary dyskinesias are primarily the result rather than the cause of chronic sinusitis. Patients with chronic sinusitis of uncertain origin exhibit a prominent loss of differentiated epithelial cells, as well as ciliary defects, most of which are likely to be secondary to the chronic disease process. These changes slow down mucociliary clearance and lead to a vicious cycle leading to chronicity.     

Primary and secondary ciliary dyskinesia

It has recently been shown that patients with Kartagener's triad and also some subjects with similar symptoms, but without situs inversus, have a congenital abnormality of cilia as an explanation for their chronic airway symptoms; this disease has been named "the immotile-cilia syndrome" or more correctly "primary ciliary dyskinesia". Studying 27 such patients, we have found daily nose blowings since birth, chronic-recurrent sinusitis, and chronic secretory otitis media highly characteristic features. The frequency of common colds was not increased, and most patients did not suffer from repeated episodes of acute purulent otitis media. The number of ciliated cells with immotile cilia was increased, but only a single patient had completely immotile cilia; also the degree of asynchrony within the single ciliated cell was increased. Electron microscopy showed a decreased number of dynein arms in some patients, and abnormal arrangement of microtubules in others. Some patients, however, had normal ultrastructure, and this appeared to be associated with a hyperfrequent beating pattern. At least three subgroups of patients with primary ciliary dyskinesia can be delineated based on the motility-ultrastructure studies. Bacterial infection tends to reduce the number of ciliated cells with motile cilia, and viral infection (common cold) gives a very marked and long-lasting reduction in the number of ciliated cells. This may account for some otherwise unexplainable subchronic symptoms from nose and throat.     

Ciliary ultrastructure and nasal mucociliary clearance in chronic and allergic rhinitis

The authors have studied nasal specimens collected by means of nasal brushing in eight patients affected by allergic rhinitis and in eight affected by chronic rhinitis, while in other four patients affected by allergic rhinitis a lower turbinate biopsy was performed. All twenty patients showed an increased mucociliary clearance time and a reduced velocity regardless to the pathology during a previously performed saccharin test. Different ultrastructural alterations have been observed, such as: both central and peripheral microtubules alterations; absence of dynein arms; absence of radial spokes; ciliary membrane alterations; "compound" cilia; disorientation of central tubules. These alterations have been observed variously associated in both allergic and chronic rhinitis patients groups. Basing on their data, the authors state that ciliary abnormalities cannot be considered specific of a particular pathology but they can coexist in different situations. They also think that the mucociliary clearance parameters determination represents the only method to evaluate, even if in an indirect fashion, the percentage of ciliary abnormalities, as no direct quantitative method has been described. Ciliary ultrastructural alterations can be of diagnostic value only if associated with mucociliary clearance time and velocity determination.     

Observation of nasal mucosal cilia ultrastructure of Kartagener's syndrome]

OBJECTIVE: To report the ultrastructure of nasal mucosal cilia of Kartagener's syndrome and the operative results of 2 cases. METHODS: Both two cases were underwent endoscopic sinus surgery. Transmission and scanning electron microscopic examinations of nasal mucosa for case 1 were performed. RESULTS: All 2 cases presented situs inversus, chronic paranasal sinusitis and bronchiectasis. The electron microscopic examination showed that the shape of nasal cilia and "9 + 2" structure were normal, but the lack of inner dynein arm of cilia and confused arrangement of central pair of microtubules were confirmed. The recurrent of nasal polyps after 1-year follow-up was observed in case 1. CONCLUSIONS: The disturbance of mucociliary clearance due to congenital ciliary structure defect might be the cause of chronic sinusitis and polyps, which might be one of reasons for poor prognosis after operation and we should pay more close attention.     

Kartagener综合征鼻窦炎的治疗--附1例报道及文献复习

目的 探讨Kartagener综合征鼻窦炎的有效治疗措施。方法 回顾诊治的1例Kartagener综合征卑窦炎病人并结合文献复习，分析Kartagener综合征及其鼻窦炎的病因、诊断和治疗方法。结果 Kartagener综合正病人因基因突变致纤毛结构和功能异常，从而出现慢性鼻窦炎、支气管扩张、内脏反位三联症，针对其鼻窦炎的肓效治疗可能可以减轻支气管、肺疾病的发展。结论 鼻窦炎常为Kartagener综合征首发症状，及早确诊并选择最合适的治疗方式，有益于疾病的转归。     

Cilia ultrastructure in children with Down syndrome

Chronic sinusitis, otitis media with effusion, and upper respiratory tract infections are commonly found in patients with Down syndrome. These diseases are generally felt to be secondary to depressed immune function and altered craniofacial dimensions. Recently, a cilia ultrastructure abnormality was found in a child with Down syndrome. This study is the first to be carried out to determine if cilia ultrastructure abnormalities are prevalent in the population with Down syndrome. Four of 10 patients had documented cilia abnormalities, but these were present in the background of normal cilia, suggesting that they were the result rather than the cause of chronic sinusitis. Similarly, nasal epithelium metaplasia was detected in 50% of the patients. Chronic sinusitis, otitis media with effusion, and recurrent upper respiratory tract infections in children with Down syndrome cannot generally be attributed to primary cilia ultrastructure abnormalities.