Ultrastructural Abnormalities of Respiratory Cilia: A 25-Year Experience

2008 ◽  
Vol 132 (11) ◽  
pp. 1786-1791
Author(s):  
Thomas P. Plesec ◽  
Angela Ruiz ◽  
James T. McMahon ◽  
Richard A. Prayson
Keyword(s):  
Basal Body ◽  
Respiratory Infections ◽  
Clinical History ◽  
Radial Spoke ◽  
Transmission Electron ◽  
Respiratory Cilia ◽  
Tract Infections ◽  
Multiple Abnormalities ◽  
Ciliary Dyskinesia ◽  
Ciliary Ultrastructure

Abstract Context.—Ciliary dyskinesia is a rare, but significant, cause of chronic respiratory infections, and transmission electron microscopy is a critical adjunct to making the diagnosis. Objective.—To investigate a single institution's experience with patients demonstrating abnormal ciliary ultrastructure. Design.—Retrospective clinicopathologic review of 278 bronchial or nasal turbinate brushings or biopsies from 1983 through 2007. Results.—There were 12 women and 9 men (mean age, 19.6 years; range, 1–54 years) with abnormal ciliary ultrastructure. Clinical history was unavailable in 3 patients, 15 (83%) of 18 patients presented with chronic or recurrent upper respiratory infections, and 3 (17%) presented with infertility. Seven (39%) of 18 patients had findings of Kartagener syndrome with situs inversus, dextrocardia, and bronchiectasis. Truncation or absence of inner or outer dynein arms occurred in 15 (71%) of 21 cases, and 5 (24%) revealed transposition defects with displacement of the central microtubules and peripheral doublets in 9 + 0 and 8 + 1 patterns. Radial spoke defects with microtubular disarray occurred in 4 (19%) of 21 cases. Compound cilia with multiple axonemes within a single outer sheath and supernumerary microtubules each occurred in 2 (10%) of the cases. Random ciliary orientation was also found in 2 (10%) of the cases, and dense granular basal body inclusions occurred in 1 case (5%). Multiple abnormalities occurred in 6 (29%) of the 21 cases. Conclusions.—Most patients presented with chronic respiratory tract infections or infertility. Dynein arm defects, transposition defects, and radial spoke defects were the most commonly encountered abnormal findings. Less-frequent abnormal findings included compound cilia, supernumerary microtubules, and dense granular basal body inclusions.

scholarly journals PCD Detect: enhancing ciliary features through image averaging and classification

2020 ◽  
Vol 319 (6) ◽  
pp. L1048-L1060
Author(s):  
Amelia Shoemark ◽  
Andreia L. Pinto ◽  
Mitali P. Patel ◽  
Farheen Daudvohra ◽  
Claire Hogg ◽  
...  
Keyword(s):  
Genetic Diagnosis ◽  
Lung Damage ◽  
Chain Gene ◽  
Heavy Chain Gene ◽  
Transmission Electron ◽  
Improve Accuracy ◽  
Image Averaging ◽  
Biallelic Mutations ◽  
Ciliary Dyskinesia ◽  
Ciliary Ultrastructure

Primary ciliary dyskinesia (PCD) is an inherited disorder of the motile cilia. Early accurate diagnosis is important to help prevent lung damage in childhood and to preserve lung function. Confirmation of a diagnosis traditionally relied on assessment of ciliary ultrastructure by transmission electron microscopy (TEM); however, >50 known PCD genes have made the identification of biallelic mutations a viable alternative to confirm diagnosis. TEM and genotyping lack sensitivity, and research to improve accuracy of both is required. TEM can be challenging when a subtle or partial ciliary defect is present or affected cilia structures are difficult to identify due to poor contrast. Here, we demonstrate software to enhance TEM ciliary images and reduce background by averaging ciliary features. This includes an option to classify features into groups based on their appearance, to generate multiple averages when a nonhomogeneous abnormality is present. We validated this software on images taken from subjects with well-characterized PCD caused by variants in the outer dynein arm (ODA) heavy chain gene DNAH5. Examining more difficult to diagnose cases, we detected 1) regionally restricted absence of the ODAs away from the ciliary base, in a subject carrying mutations in DNAH9; 2) loss of the typically poorly contrasted inner dynein arms; and 3) sporadic absence of part of the central pair complex in subjects carrying mutations in HYDIN, including one case with an unverified genetic diagnosis. We show that this easy-to-use software can assist in detailing relationships between genotype and ultrastructural phenotype, improving diagnosis of PCD.

Incidence of Primary Ciliary Dyskinesia in Children with Recurrent Respiratory Diseases

1997 ◽  
Vol 106 (10) ◽  
pp. 854-858 ◽  
Author(s):  
André Coste ◽  
Marie-Claude Millepied ◽  
Catherine Chapelin ◽  
Philippe Reinert ◽  
Françoise Poron ◽  
...  
Keyword(s):  
Primary Ciliary Dyskinesia ◽  
Respiratory Tract Infections ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Systematic Investigation ◽  
Ciliated Cells ◽  
Tract Infections ◽  
Ciliary Dyskinesia ◽  
Ciliary Ultrastructure ◽  
Recurrent Respiratory Tract Infections

The goal of the study was to evaluate the incidence of primary ciliary dyskinesia (PCD) in children suffering from recurrent respiratory tract infections (RRIs) by means of a noninvasive method. Respiratory ciliated cells were collected by nasal brushing in 118 children (4.6 ± 2.5 years) with RRIs. The ciliary beat frequency (CBF) was measured with a stroboscopic method, and when the CBF was abnormal, the ciliary ultrastructure was analyzed by a quantitative method. The CBF could be measured in 106 patients (90%) and was abnormal in 15 patients. The ciliary ultrastructure was found to be abnormal in 11 of 15 patients: PCD was diagnosed in 6 cases, and acquired ciliary defects were observed in the remaining 5 patients. Our conclusion, that PCD is rare but not exceptional (5.6%) in children with RRIs, justifies the systematic investigation of ciliated cells in such patients. For this purpose, nasal brushing can be used to sample ciliated cells even in young children.

scholarly journals Structural insights into the cause of human RSPH4A primary ciliary dyskinesia

2021 ◽  
pp. mbc.E20-12-0806
Author(s):  
Yanhe Zhao ◽  
Justine Pinskey ◽  
Jianfeng Lin ◽  
Weining Yin ◽  
Patrick R. Sears ◽  
...  
Keyword(s):  
Primary Ciliary Dyskinesia ◽  
Electron Tomography ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Structural Basis ◽  
Radial Spoke ◽  
Radial Spokes ◽  
Cilia And Flagella ◽  
Tract Infections ◽  
Ciliary Dyskinesia

Cilia and flagella are evolutionarily conserved eukaryotic organelles involved in cell motility and signaling. In humans, mutations in Radial Spoke Head Protein 4 homolog A ( RSPH4A) can lead to primary ciliary dyskinesia (PCD), a life-shortening disease characterized by chronic respiratory tract infections, abnormal organ positioning, and infertility. Despite its importance for human health, the location of RSPH4A in human cilia has not been resolved, and the structural basis of RSPH4A-/- PCD remains elusive. Here, we present the native, three-dimensional structure of RSPH4A-/- human respiratory cilia using samples collected non-invasively from a PCD patient. Using cryo-electron tomography and subtomogram averaging, we compared the structures of control and RSPH4A-/- cilia, revealing primary defects in two of the three radial spokes (RSs) within the axonemal repeat and secondary (heterogeneous) defects in the central pair complex. Similar to RSPH1-/- cilia, the radial spoke heads of RS1 and RS2, but not RS3, were missing in RSPH4A-/- cilia. However, RSPH4A-/- cilia also exhibited defects within the arch domains adjacent to the RS1 and RS2 heads, which were not observed with RSPH1 loss. Our results provide insight into the underlying structural basis for RSPH4A-/- PCD and highlight the benefits of applying cryo-ET directly to patient samples for molecular structure determination. [Media: see text]

scholarly journals Frequency and Types of Ciliary Ultrastructural Defects in Patients With Suspected Primary Ciliary Dyskinesia Symptoms Analyzed by Transmission Electron Microscopy

2021 ◽  
Author(s):  
Mitra Rezaei ◽  
Amirali Soheili ◽  
Atefeh Fakharian ◽  
Hamid Jamaati ◽  
Jahangir Ghorbani ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Primary Ciliary Dyskinesia ◽  
Respiratory Infections ◽  
Descriptive Analysis ◽  
Final Diagnosis ◽  
International Consensus ◽  
Tem Analysis ◽  
Transmission Electron ◽  
Ciliary Dyskinesia

Abstract Background: Primary ciliary dyskinesia (PCD) is a rare autosomal recessive condition of often chronic respiratory infections in early life. A useful tool for early diagnosis of such ciliary abnormalities is transmission electron microscopy (TEM). This study aimed to use TEM to examine these defects and speculate on a diagnosis.Methods: From 2017 to 2019, all referral patients with suspected PCD symptoms were included in this study. Nasal samples were taken after exclusion of further potential differential diagnosis and prepared for TEM. The final diagnosis was based on the International Consensus Guideline for reporting transmission electron microscopy results in the diagnosis of PCD. A descriptive analysis of demographic and ciliary ultrastructural data was performed by SPSS ver 21.Results: Study population consisted of 37 women and 30 men (mean age=20.34±10.7 years). The clinical presentations were as follows: bronchiectasis: 26 patients (38.8%); sinusitis: 23(34.3%); recurrent respiratory infection: 21 patients (31.3%); auditory symptoms: 5 patients (7.5%); situs inversus: 3 patients (4.4%); productive cough: 2 patients (3%); infertility: 2 patients (3%); polyposis: 1 patient (1.5%). According to TEM analysis, 12 (17%) of patients were PCD, 11 (15.7%) were indicating PCD cases, 26 (37.1%) of them had no criteria of PCD and 18 (25.7%) of cases had normal ciliary ultrastructure. Compound cilia and extra-tubule were reported in 29 (41.4%) and 31(44.3%) of patients, respectively. The outer dynein arm defect was seen in 11(16.4%) cases and the inner dynein arm (IDA) defect was seen in 20 (29.8%) cases. Two patients (3%) had microtubular disorganization.Conclusion: Bronchiectasis and sinusitis were the most common complications. The compound cilia and extra-tubule were the most prevalent TEM finding among all participants. However, the most prevalent hallmark diagnostic defects among PCD patients were ODA and IDA defects among PCD patients. Other diagnostic PCD tests should also be performed in patients in the indicating PCD group, those without PCD criteria, and normal patients with a highly suggestive history. Cell-culture, as well, should confirm IDA defects. This study highlights the fundamental need to consider ciliary defect among probable diagnoses and use TEM as a practical diagnostic tool.

scholarly journals Ciliary Feature Counter: A program for the Quantitative Assessment of Cilia to Diagnose Primary Ciliary Dyskinesia

Diagnostics ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 524
Author(s):  
Andreia L. Pinto ◽  
Ranjit K. Rai ◽  
Claire Hogg ◽  
Thomas Burgoyne
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Primary Ciliary Dyskinesia ◽  
Quantitative Assessment ◽  
Consensus Guideline ◽  
International Consensus ◽  
Transmission Electron ◽  
Speed Up ◽  
Ciliary Dyskinesia ◽  
Ciliary Ultrastructure

Primary ciliary dyskinesia (PCD) is a disorder that affects motile cilia in the airway that are required for the removal of mucus, debris, and pathogens. It is important to diagnose PCD in early childhood to preserve lung function. The confirmation of a diagnosis relies on the assessment of ciliary ultrastructure by transmission electron microscopy (TEM). TEM involves the quantitative assessment of the ciliary ultrastructure to identify PCD defects as well as abnormalities resulting from infection. Many specialist diagnostic centres still rely on physical counters to tally results and paper notes to summarise findings before transferring the results to computer databases/records. To speed up the diagnostic data collection and increase the protection of patient information, we have developed digital ciliary feature counters that conform to the PCD reporting international consensus guideline. These counters can be used on a computer or tablet, and automatically generate notes regarding sample observations. We show that the digital counters are easy to use and can generate TEM diagnostic reports that will be useful for many PCD diagnostic centres.

scholarly journals PRIMARY CILIARY DYSKINESIA IMMOTILE CILIA SYNDROME

2019 ◽  
Vol 20 (4) ◽  
pp. 237-245
Author(s):  
Yulia A. Tsareva ◽  
N. I. Zryachkin ◽  
M. A. Kuznetsova
Keyword(s):  
Primary Ciliary Dyskinesia ◽  
High Speed ◽  
Hereditary Disease ◽  
Screening Tests ◽  
Cell Culture Study ◽  
Immotile Cilia ◽  
Transmission Electron ◽  
Tract Infections ◽  
Ciliary Dyskinesia

Primary ciliary dyskinesia (PCD) is a genetically heterogeneous hereditary disease characterized by recurrent respiratory tract infections, decreased fertility, and situs inversus in 50% of cases. The core of the syndrome is the disturbance of mucociliary clearance due to the lack or defect of cilia leading to their partial or complete immobility. There are some tests for diagnostic PCD with specific benefits and limitations, but there is still no diagnostic «gold standard» yet. Identification of nitric oxide and nasal clearance of dye or saccharin are widely used as screening tests. Clearance of 99Tc-labeled colloidal albumin, high-speed video microscopy and transmission electron microscopy, the cell culture study and genetic testing are methods for the verification. Late identification of PCD is reported worldwide. There are no methods to control the development of PCD complications. The important role is played by the long-term and constant follow up (including spirometry, evaluation of pulmonary clearance and X-ray scanning).

scholarly journals Primary ciliary dyskinesia in a Staffordshire bull terrier : clinical communication

2004 ◽  
Vol 75 (3) ◽  
Author(s):  
M. De Scally ◽  
R.G. Lobetti ◽  
E. Van Wilpe
Keyword(s):  
Cross Sections ◽  
Primary Ciliary Dyskinesia ◽  
Clinical Communication ◽  
First Case ◽  
Transmission Electron ◽  
Radial Spokes ◽  
History Of ◽  
Tract Infections ◽  
Ciliary Dyskinesia ◽  
Recurrent Respiratory Tract Infections

Primary ciliary dyskinesia (PCD) is a diverse group of inherited structural and functional abnormalities of the respiratory and other cilia, which results in recurrent respiratory tract infections. Primary ciliary dyskinesia was diagnosed in a 14-week old Staffordshire bull terrier that had a history of respiratory disease from 7 weeks of age. Pneumonia was diagnosed on thoracic radiographs and transtracheal aspirate. Transmission electron microscopy of the bronchi and trachea indicated the presence of both primary and secondary ciliary dyskinesia. The most prominent primary defects consisted of absent inner dyneim arms, absent radial spokes and absence of the central microtubules. These defects accounted for 62 % of the total number of cross-sections screened. Non-specific ciliary abnormalities encountered most often were compound cilia, swollen cilia, addition / deletion of peripheral doublets and disorganised axonemes (26 %). To the authors' knowledge, this is the first case of PCD described in the Staffordshire bull terrier and the first report of PCD in South Africa.

scholarly journals Quantitative Assessment of Ciliary Ultrastructure with the Use of Automatic Analysis: PCD Quant

Diagnostics ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1363
Author(s):  
Andrea Felšöová ◽  
Tibor Sloboda ◽  
Lukáš Hudec ◽  
Miroslav Koblížek ◽  
Petr Pohunek ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Automatic Analysis ◽  
Diagnostic Methods ◽  
Mutual Orientation ◽  
Transmission Electron ◽  
Complex Process ◽  
Golden Standard ◽  
Secondary Defect ◽  
Ciliary Dyskinesia ◽  
Ciliary Ultrastructure

The ciliary ultrastructure can be damaged in various situations. Such changes include primary defects found in primary ciliary dyskinesia (PCD) and secondary defects developing in secondary ciliary dyskinesia (SCD). PCD is a genetic disease resulting from impaired ciliary motility causing chronic disease of the respiratory tract. SCD is an acquired condition that can be caused, for example, by respiratory infection or exposure to tobacco smoke. The diagnosis of these diseases is a complex process with many diagnostic methods, including the evaluation of ciliary ultrastructure using transmission electron microscopy (the golden standard of examination). Our goal was to create a program capable of automatic quantitative analysis of the ciliary ultrastructure, determining the ratio of primary and secondary defects, as well as analysis of the mutual orientation of cilia in the ciliary border. PCD Quant, a program developed for the automatic quantitative analysis of cilia, cannot yet be used as a stand-alone method for evaluation and provides limited assistance in classifying primary and secondary defect classes and evaluating central pair angle deviations. Nevertheless, we see great potential for the future in automatic analysis of the ciliary ultrastructure.

Sign in / Sign up

Export Citation Format

Share Document