Ciliary Pattern Research Articles

Impact of local anesthesia on ciliary dyskinesia diagnosis by digital high-speed videomicroscopy.

It was hypothesized that local anesthesia administration would not significantly affect ciliary function in nasal epithelium. A prospective, simple-blind randomized study was conducted between 2020 and 2023. The study employed digital high-speed videomicroscopy to analyze ciliary beat frequency and pattern. A cohort of 38 participants was recruited, consisting of 25 healthy volunteers and 13 referred individuals (including seven diagnosed with PCD). Selection criteria ensured the absence of chronic respiratory diseases, recent respiratory tract infections, or regular use of nasal medications. Participants underwent nasal brushing with administration of lidocaine and naphazoline nasal spray in one nostril and saline in the contralateral nostril. Ciliary beat frequency and pattern were measured using digital high-speed video microscopy. Nasal spray administration did not significantly alter ciliary beat frequency or pattern compared to saline (p = 0.841 and p = 0.125, respectively). Subgroup analysis revealed consistent results across healthy volunteers, referred patients, and PCD patients. Local anesthesia with lidocaine and naphazoline spray did not affect ciliary function outcomes. These findings support the safe use of these agents in clinical practice for PCD diagnostic procedures. Further research with larger cohorts is warranted for validation.

Morphological and molecular investigation of Scolelepis agilis (Verrill, 1873) (Annelida: Spionidae) from beaches of the Atlantic coast of North America

Polychaete worms of the genus Scolelepis are one of the most abundant members of beach ecosystems worldwide. Along the Atlantic coast of North America, specimens of Scolelepis inhabiting high-energy beaches have long been erroneously identified as the European species Scolelepis squamata (O.F. Müller, 1806). In this study, we examined specimens of Scolelepis collected from Massachusetts to Virginia, finding they match Scolelepis agilis (Verrill, 1873), originally described from beaches in New Jersey. Scolelepis agilis can be most easily distinguished from S. squamata based on differences in palp ciliary pattern, morphology of neuropodial lamellae, and the quantity and start of both notopodial and neuropodial hooded hooks. Live specimens of S. agilis collected from New York were also examined to provide data on their reproduction and morphometric analyses. In addition, molecular data (18S, 28S rRNA, cytochrome c oxidase I and 16S rRNA) showed that specimens of S. agilis from the north to mid-Atlantic coast formed a monophyletic group (p-distance ≤ 0.004 for MA, RI, NY and VA populations) and were distinct from S. squamata (p-distance ≥ 0.196 (0.03 SE)). The molecular data also suggest the presence of several misidentified or undescribed species of Scolelepis, highlighting the need for additional taxonomic work on the genus from these localities as well as others (e.g. South Africa) where the worms have been incorrectly identified as S. squamata. A key to species of Scolelepis from the east coast of the United States is provided and a lectotype and paralectotypes for S. agilis are herein designated.

Primary Ciliary Dyskinesia Associated Disease-Causing Variants in CCDC39 and CCDC40 Cause Axonemal Absence of Inner Dynein Arm Heavy Chains DNAH1, DNAH6, and DNAH7.

Disease-causing bi-allelic DNA variants in CCDC39 and CCDC40 are frequent causes of the hereditary disorder of primary ciliary dyskinesia (PCD). The encoded proteins form a molecular ruler complex, crucial for maintaining the 96 nm repeat units along the ciliary axonemes. Defects of those proteins cause a stiff, rapid, and flickery ciliary beating pattern, recurrent respiratory infections, axonemal disorganization, and abnormal assembly of GAS8, CCDC39, and DNALI1. We performed molecular characterization of the defects in the 96 nm axonemal ruler due to disease-causing variants in CCDC39 and CCDC40 and analyzed the effect on additional axonemal components. We identified a cohort of 51 individuals with disease-causing variants in CCDC39 and CCDC40 via next-generation sequencing techniques and demonstrated that the IDA heavy chains DNAH1, DNAH6, and DNAH7 are conspicuously absent within the respiratory ciliary axonemes by immunofluorescence analyses. Hence, we show for the first time that the centrin2 (CETN2) containing IDAs are also affected. These findings underscore the crucial role of CCDC39 and CCDC40 in the assembly and function of IDAs in human respiratory cilia. Thus, our data improve the diagnostics of axonemal ruler defects by further characterizing the associated molecular IDA defects.

Advancing Primary Ciliary Dyskinesia Diagnosis through High-Speed Video Microscopy Analysis.

Primary ciliary dyskinesia (PCD) is an inherited disorder that impairs motile cilia, essential for respiratory health, with a reported prevalence of 1 in 16,309 within Hispanic populations. Despite 70% of Puerto Rican patients having the RSPH4A [c.921+3_921+6del (intronic)] founder mutation, the characterization of the ciliary dysfunction remains unidentified due to the unavailability of advanced diagnostic modalities like High-Speed Video Microscopy Analysis (HSVA). Our study implemented HSVA for the first time on the island as a tool to better diagnose and characterize the RSPH4A [c.921+3_921+6del (intronic)] founder mutation in Puerto Rican patients. By applying HSVA, we analyzed the ciliary beat frequency (CBF) and pattern (CBP) in native Puerto Rican patients with PCD. Our results showed decreased CBF and a rotational CBP linked to the RSPH4A founder mutation in Puerto Ricans, presenting a novel diagnostic marker that could be implemented as an axillary test into the PCD diagnosis algorithm in Puerto Rico. The integration of HSVA technology in Puerto Rico substantially enhances the PCD evaluation and diagnosis framework, facilitating prompt detection and early intervention for improved disease management. This initiative, demonstrating the potential of HSVA as an adjunctive test within the PCD diagnostic algorithm, could serve as a blueprint for analogous developments throughout Latin America.

The genetic framework of primary ciliary dyskinesia assessed by soft computing analysis.

International guidelines disagree on how best to diagnose primary ciliary dyskinesia (PCD), not least because many tests rely on pattern recognition. We hypothesized that quantitative distribution of ciliary ultrastructural and motion abnormalities would detect most frequent PCD-causing groups of genes by soft computing analysis. Archived data on transmission electron microscopy and high-speed video analysis from 212 PCD patients were re-examined to quantitate distribution of ultrastructural (10 parameters) and functional ciliary features (4 beat pattern and 2 frequency parameters). The correlation between ultrastructural and motion features was evaluated by blinded clustering analysis of the first two principal components, obtained from ultrastructural variables for each patient. Soft computing was applied to ultrastructure to predict ciliary beat frequency (CBF) and motion patterns by a regression model. Another model classified the patients into the five most frequent PCD-causing gene groups, from their ultrastructure, CBF and beat patterns. The patients were subdivided into six clusters with similar values to homologous ultrastructural phenotype, motion patterns, and CBF, except for clusters 1 and 4, attributable to normal ultrastructure. The regression model confirmed the ability to predict functional ciliary features from ultrastructural parameters. The genetic classification model identified most of the different groups of genes, starting from all quantitative parameters. Applying soft computing methodologies to PCD diagnostic tests optimizes their value by moving from pattern recognition to quantification. The approach may also be useful to evaluate atypical PCD, and novel genetic abnormalities of unclear disease-producing potential in the future.

Tumor Profiles of Late Presentation Uveal Ring Melanoma With Novel Characteristics - A Case Series.

The purpose of this study was to investigate the clinical features, tumor characteristics, including histopathology and cytogenetic analysis, and management of patients with uveal ring melanoma in New Zealand. A retrospective review was conducted on all uveal melanoma cases treated in a single national oncology center in New Zealand. The study period was from January 1, 2013 to December 31, 2022 (10 years). Written consent was obtained from all patients included in this case series. Uveal ring melanoma of ciliary body origin (n = 4) comprised 0.7% of all uveal melanomas (n = 571). Ethnicity distribution was three patients of New Zealand European ancestry and one patient of Chinese/Pasifika ancestry. Three patients (75%) were symptomatic at presentation (spontaneous hyphema, glaucoma, and cataract), whereas one was asymptomatic but subsequently developed painful refractory glaucoma. All eyes underwent enucleation. Three eyes had primary iris biopsies with subsequent enucleation for refractory glaucoma and pain and one eye underwent primary enucleation. All cases demonstrated malignant tumor characteristics including diffuse 270 to 360-degree ciliary body ring growth pattern, epithelioid-cell type and presence of either BAP-1 expression loss or gain of MYC gene. Two cases (50%) developed distant organ metastasis - liver, parotid gland, and breast. Of those, one patient was deceased at the time of follow-up whereas one had completed treatment for metastases. Uveal ring melanoma is a rare tumor that is highly invasive and malignant. When detected, prompt definitive treatment should be advocated, and patient counselling should be given regarding the high risk of developing a painful, blind eye with increased risk of metastasis in the absence of treatment.

High-Speed Video Microscopy of Ependymal Cilia in Brain Organotypic and Cell Culture Models.

The wall of the ventricular system within the neuraxis is lined almost entirely by E1 ependymal cells, each of which projects multiple motile cilia from their apical surface into the cerebrospinal fluid (CSF). This specialized layer of E1 cells constitutes the border between the CSF and the brain interstitial fluid (BIF), and by controlling influx and efflux across the CSF to BIF interface, it is increasingly recognized to play an integral role in modulating and maintaining the brain microenvironment. The motile cilia have been shown to be responsive to changes in the CSF microenvironment, and while the physiological role of this mechanism remains incompletely understood, manipulating this control mechanism may influence the brain microenvironment potentially opening a new frontier in therapeutic intervention.In this paper, we describe our techniques for preparing organotypic slices from the murine brain parenchyma and establishing cell cultures of multiciliated ependymal cells from mouse and rat neonatal brain tissue. Our methodology generates a functional readout of ciliary function, specifically high-speed video microscopy (HSVM) enables the quantification of ciliary beat frequency (CBF), and characterization of ciliary beat pattern.

Temporal Stability of Ciliary Beating Post Nasal Brushing, Modulated by Storage Temperature.

Primary ciliary dyskinesia is a heterogeneous, inherited motile ciliopathy in which respiratory cilia beat abnormally, and some ultrastructural ciliary defects and specific genetic mutations have been associated with particular ciliary beating alterations. Ciliary beating can be evaluated using digital high-speed videomicroscopy (DHSV). However, normal reference values, essential to assess ciliary beating in patients referred for a PCD diagnostic, vary between centres, as minor variations in protocols might influence ciliary beating. Consequently, establishment of normal values is essential for each PCD diagnostic centre. We aimed to evaluate whether delay after sampling, and temperature for conservation of respiratory ciliated samples, might modify assessments of ciliary beating. In total, 37 healthy nasal brushing samples of respiratory ciliated epithelia were collected. Video sequences were recorded at 37 °C immediately using DHSV. Then, the samples were divided and conserved at 4 °C or at room temperature (RT). Ciliated beating edges were then recorded at 37 °C, at 3 h and at 9 h post sampling. In six samples, recordings were continued up to 72 h after sampling. Ciliary beating was assessed manually by ciliary beat frequency (CBFM) and ciliary beat pattern (CBP). A semi-automatic software was used for quantitative analysis. Both CBF and CBP evaluated manually and by a semi-automated method were stable 9 h after sampling. CBFM was higher when evaluated using samples stored at RT than at 4 °C. CBP and the semi-automated evaluation of ciliary beating were not affected by storage temperature. When establishing normal references values, ciliary beating can be evaluated at 37 °C up to 9 h after nasal brushing, but the storage temperature modifies ciliary beating and needs to be controlled.

New evidence of consistency between phylogeny and morphology for two taxa in ciliated protists, the subclasses Oligotrichia and Choreotrichia (Protista, Ciliophora)

Marine planktonic ciliates are largely oligotrichs and choreotrichs, which are two subclasses of the class Spirotrichea. The current phylogenetic assignments of oligotrichs and choreotrichs are inconsistent with previous results based on morphological features, probably hindered by the limited information from a single gene locus. Here we provide 53 new sequences from small subunit ribosomal RNA (SSU rDNA), ITS1-5.8S rDNA-ITS2, and large subunit ribosomal RNA (LSU rDNA) gene loci in 25 oligotrich and choreotrich species. We also predict RNA secondary structures for the ITS2 regions in 55 species, 48 species of which are reported for the first time. Based on these novel data, we make a more comprehensive phylogenetic reconstruction, revealing consistency between morphological taxonomy and an updated phylogenetic system for oligotrichs and choreotrichs. With the addition of data from ciliature patterns and genes, the phylogenetic analysis of the subclass Oligotrichia suggests three evolutionary trajectories, among which: 1) Novistrombidium asserts an ancestral ciliary pattern in Oligotrichia; 2) the subgenera division of Novistrombidium and Parallelostrombidium are fully supported; 3) the three families (Tontoniidae, Pelagostrombidiidae and Cyrtostrombidiidae) all evolved from the most diverse family Strombidiidae, which explains why strombidiids consistently form polyphyletic clades. In the subclass Choreotrichia, Strombidinopsis likely possesses an ancestral position to other choreotrichs, and both phylogenetic analysis and RNA secondary structure prediction support the hypothesis that tintinnids may have evolved from Strombidinopsis. The results presented here offer an updated hypothesis for the evolutionary history of oligotrichs and choreotrichs based on new evidence obtained by expanding sampling of molecular information across multiple gene loci.

The Cilialyzer – A freely available open-source software for the analysis of mucociliary activity in respiratory cells

Background and ObjectivePrimary ciliary dyskinesia (PCD) is a rare genetic disorder causing a defective ciliary structure, which predominantly leads to an impaired mucociliary clearance and associated airway disease. As there is currently no single diagnostic gold standard test, PCD is diagnosed by a combination of several methods comprising genetic testing and the examination of the ciliary structure and function. Among the approved diagnostic methods, only high-speed video microscopy (HSVM) allows to directly observe the ciliary motion and therefore, to directly assess ciliary function. In the present work, we present our recently developed freely available open-source software – termed “Cilialyzer”, which has been specifically designed to support and facilitate the analysis of the mucociliary activity in respiratory epithelial cells captured by high-speed video microscopy. MethodsIn its current state, the Cilialyzer software enables clinical PCD analysts to load, preprocess and replay recorded image sequences as well as videos with a feature-rich replaying module facilitating the commonly performed qualitative visual assessment of ciliary function (including the assessment of the ciliary beat pattern). The image processing methods made accessible through an intuitive user interface allow clinical specialists to comfortably compute the ciliary beating frequency (CBF), the activity map and the “frequency correlation length” – an observable getting newly introduced. Furthermore, the Cilialyzer contains a simple-to-use particle tracking interface to determine the mucociliary transport speed. ResultsCilialyzer is fully written in the Python programming language and freely available under the terms of the MIT license. The proper functioning of the computational analysis methods constituting the Cilialyzer software is demonstrated by using simulated and representative sample data from clinical practice. Additionally, the software was used to analyze high-speed videos showing samples obtained from healthy controls and genetically confirmed PCD cases (DNAI1 and DNAH11 mutations) to show its clinical applicability. ConclusionsCilialyzer serves as a useful clinical tool for PCD analysts and provides new quantitative information awaiting to be clinically evaluated using cohorts of PCD. As Cilialyzer is freely available under the terms of a permissive open-source license, it serves as a ground frame for further development of computational methods aiming at the quantification and automation of the analysis of mucociliary activity captured by HSVM.

Cinetochilides minimus sp. nov., a Tiny Benthic Ciliate (Protozoa, Ciliophora) from Brackish Water in Korea

During a field survey of Korean marine and brackish water ciliate diversity, we collected a tiny benthic ciliate (13–18 µm long in vivo) from the opening of a brackish water lagoon (10‰). At low magnification, it resembles members of the genus Aspidisca because of the oval body shape and the benthic life style, but is not thigmotactic. Based on the observations of living cells, silver-impregnated specimens (i.e., using protargol, silver carbonate, and wet silver nitrate), SEM images, and the 18S rRNA gene sequences, we confirmed that it is a new member of the genus Cinetochilides. The new species, C. minimus sp. nov., can be easily distinguished from other congeners mainly by the fragmented somatic kinety 1. In spite of the small size, the new species has more than 200 basal bodies, including those in the oral apparatus. The arrangement of the ciliary pattern is rather confusing because of the polymerized kinetids, the sparse basal bodies, the non-ciliated area on the dorsal side, and the presence of parasomal sacs next to the kinetosomes. In the present study, we provide a detailed morphological description and infer the phylogenetic position of Cinetochilides minimus sp. nov. based on 18S rRNA gene sequences.

Light-sheet laser speckle imaging for cilia motility assessment

Mucociliary clearance is an important innate defense mechanism predominantly mediated by ciliated cells in the upper respiratory tract. Ciliary motility on the respiratory epithelium surface and mucus pathogen trapping assist in maintaining healthy airways. Optical imaging methods have been used to obtain several indicators for assessing ciliary movement. Light-sheet laser speckle imaging (LSH-LSI) is a label-free and non-invasive optical technique for three-dimensional and quantitative mapping of velocities of microscopic scatterers. Here, we propose to use an inverted LSH-LSI platform to study cilia motility. We have experimentally confirmed that LSH-LSI can reliably measure the ciliary beating frequency and has the potential to provide many additional quantitative indicators for characterizing the ciliary beating pattern without labeling. For example, the asymmetry between the power stroke and the recovery stroke is apparent in the local velocity waveform. PIV (particle imaging velocimetry) analysis of laser speckle data could determine the cilia motion directions in different phases.

Basal bodies bend in response to ciliary forces.

Motile cilia beat with an asymmetric waveform consisting of a power stroke that generates a propulsive force and a recovery stroke that returns the cilium back to the start. Cilia are anchored to the cell cortex by basal bodies (BBs) that are directly coupled to the ciliary doublet microtubules (MTs). We find that, consistent with ciliary forces imposing on BBs, bending patterns in BB triplet MTs are responsive to ciliary beating. BB bending varies as environmental conditions change the ciliary waveform. Bending occurs where striated fibers (SFs) attach to BBs and mutants with short SFs that fail to connect to adjacent BBs exhibit abnormal BB bending, supporting a model in which SFs couple ciliary forces between BBs. Finally, loss of the BB stability protein Poc1, which helps interconnect BB triplet MTs, prevents the normal distributed BB and ciliary bending patterns. Collectively, BBs experience ciliary forces and manage mechanical coupling of these forces to their surrounding cellular architecture for normal ciliary beating.

Taxonomy and molecular phylogeny of two new Blepharisma species (Ciliophora, Heterotrichea) from northeastern China

The morphology and ciliary pattern of two new heterotrich ciliates, Blepharisma orientale n. sp. and B. sinicum n. sp., collected from Sanjiang Plain Wetland, northeastern China, were investigated using living observation and protargol staining. Blepharisma orientale n. sp. is characterized by the following combination of characteristics: limnetic; body in vivo 280–380 × 120–190 μm, slender to elongate, cortical granules dark pink, ventral margin somewhat sigmoid; buccal field about 55–60% of body length; 30–39 somatic kineties; 70–76 adoral membranelles, each comprising two long rows of basal bodies; paroral membrane consists of dikinetids with the posterior third obliquely arranged, both basal bodies in each dikinetid are ciliated. Blepharisma sinicum n. sp. is recognized by the combination of the following characters: limnetic; body 190–260 × 100–170 μm in size in vivo, irregularly sigmoid and slightly bilaterally flattened, dorsal margin curved; cortical granules pale pink in color; buccal field about 60–75% of body length; 25–29 somatic kineties; 53–82 adoral membranelles; paroral membrane consisting entirely of dikinetids. Phylogenetic analyses based on small subunit rRNA (SSU rRNA) gene sequence data recovered the two new species as sister taxa, both the sister group of B. penardi.

Non-typeable Haemophilus influenzae biofilm formation on primary human airway epithelia cultured at air-liquid interface

Introduction Primary ciliary dyskinesia (PCD) is an inherited heterogeneous disorder associated with defective motile cilia on airway epithelia, resulting in impaired mucociliary function and ultimately, bronchiectasis. Non-typeable Haemophilus influenzae (NTHi) is frequently isolated from PCD airways where it resides in biofilms. It is unclear if biological pathways in response to NTHi infection differ in PCD, contributing to increased susceptibility to biofilm-associated infection. Aim The development of a biologically representative co-culture model to investigate the interactions between PCD airway epithelia and NTHi biofilms. Methods Air-liquid interface (ALI)-cultured non-PCD primary nasal cells were grown in transwells. Barrier properties were assessed by trans-epithelial electrical resistance (TEER), FITC-dextran passage and tight junction staining. Ciliary function was assessed by high-speed video microscopy at 37°C. ALI cultures were infected with a PCD NTHi isolate at multiplicity of infections (MOIs) 10, 50 or 100 and co-cultured to form biofilms over 3 days. TEER was measured pre- and post-infection, NTHi recoverability assessed by conventional culture, and biofilm formation confirmed by scanning electron microscopy (SEM). Results Epithelial culture at ALI was successful, with tight barriers being formed and normal ciliary beat pattern and frequency recorded (mean 14Hz). Following co-culture, TEER increased relative to pre-infection at all MOIs but maximum NTHi recoverability was at MOI 50 (median colony forming units, 6.8x106per cm2). SEM confirmed NTHi biofilm formation at MOI 50. Conclusions A successful NTHi biofilm co-culture model on human primary epithelia was established. This will be used with PCD epithelia to investigate host-pathogen interactions during NTHi biofilm colonisation.

Taxonomy and phylogeny of the freshwater tintinnid Tintinnopsis tubuformis Chiang, 1956 (Ciliophora, Oligotrichea) and a proposed synonymization of T. longa nom. corr. Chiang, 1956.

Tintinnid ciliates are traditionally identified by their loricae; however, increasing evidence indicates that some lorica features (e.g. its length, spiraled structures) are not reliable. The vast majority of tintinnids inhabit the marine pelagial; merely, about thirty species live in freshwater. In the present study, two morphotypes with similar lorica shapes and opening diameters but deviating lorica lengths were isolated from freshwater samples collected at different water temperatures near Chongming Island in the Yangtze Estuary, China. The specimens were studied in vivo and after protargol staining, and their phylogenetic placement was inferred from three ribosomal RNA markers; further, cell division was investigated in the short morphotype. Based on the original descriptions, the longer morphotype is identified as Tintinnopsis longa nom. corr. Chiang, 1956, and the shorter one as Tintinnopsis tubuformis Chiang, 1956. Despite distinct differences in the lorica lengths, the identity of the three molecular markers in both morphotypes suggests conspecificity, which is supported by overlapping ranges in the lorica opening diameters and the length-independent features of the somatic ciliary pattern (e.g. number of kineties). Hence, we synonymized T. longa nom. corr. with T. tubuformis and neotypified the later species.

Updating the phylogeny and taxonomy of pleurostomatid ciliates (Protista: Ciliophora) with establishment of a new family, a new genus and two new species

AbstractThe order Pleurostomatida is one of the most confusing groups in the ciliate class Litostomatea. In the present study, two new species, Paralitonotus foissneri gen. & sp. nov. and Loxophyllum apochlorelligerum sp. nov., were investigated using an integrative approach. Paralitonotus foissneri differs from other pleurostomatids by its distinctive ciliary pattern (i.e. the anterior ends of the right somatic kineties are progressively shortened from the middle towards dorsal and perioral ones). Therefore, we place it in the new genus Paralitonotus. Loxophyllum apochlorelligerum is characterized by seven to nine left and 12–14 right somatic kineties, three to five anterior dorsal warts and a subterminal contractile vacuole. Phylogenetic analyses show that Pa. foissneri clusters with Apolitonotus lynni and Pseudolitonotus gracilis to form a deep-branching subclade that is basal to other core pleurostomatids. All three genera have distinctive anteriorly shortened right kineties, suggesting that this might be a familial character state. We therefore establish a new family, Paralitonotidae fam. nov., for pleurostomatids with this character and propose a new taxonomic scheme and a hypothetical evolutionary trajectory of the order Pleurostomatida. In addition, the generic diagnostic characters for pleurostomatids are updated, and a key to the identification of pleurostomatid genera is provided.

Taxonomy and Phylogeny of Two Tintinnid Ciliates of Leprotintinnus (Protista, Ciliophora, Choreotrichida) Combining the Loricae, Cytological, Ontogenetic Features, and Barcoding Genes

Tintinnid ciliates are a highly diverse and essential group in the marine planktonic microbial loop. However, most of the known tintinnids were recorded only by the lorica characters and very few of them had been studied on their cytological features. In this study, the morphological characters of the lorica, ciliary pattern, nuclear apparatus, ontogenesis, and the molecular phylogeny of two poorly known tintinnid ciliates,Leprotintinnus nordqvisti(Brandt, 1906)Kofoid and Campbell (1929)andL. simplexSchmidt (1902), isolated from coastal waters of southern China, were investigated based on living observation, silver staining, three nuclear ribosomal DNA markers (18S, ITS1-5.8S-ITS2, and 28S genes) and one mitochondrial DNA marker (CO1gene). For the first time, the somatic ciliary pattern of the genusLeprotintinnuswas disclosed,viz., comprising a ventral, a dorsal, and a posterior kinety as well as a right, a left, and a lateral ciliary field. The diagnoses of bothLeprotintinnusspecies were improved and the neotype was assigned. The ontogenesis ofL. nordqvistiwas in enantiotropic division mode with the new dorsal and posterior kineties generatedde novo. The molecular phylogeny confirmed thatLeprotintinnusspecies are closely related to some species ofTintinnopsis,Stylicauda,Rhizodomus, andClimacocylis. The anterior extending of the ventral kinety together with some of the lateral kinety is likely to be a distinguishing feature to determine their systematic relationships. This study also revealed that (i) the lorica ofL. nordqvistiis polymorphic or plastic; (ii)Leprotintinnus tubulosusRoxas (1941)might be a synonym ofL.nordqvisti; (iii)Leprotintinnus neriticussensuYoo et al. (1988)might be a misidentification ofL. simplex.

Study of the ciliary function in adult and pediatric patients with cystic fibrosis

Mucociliary clearance MCC) results in an effective interaction between the mucus layer and the normal coordinated ciliary beating. In cystic fibrosis (CF), MCC is impaired due to dehydrated mucus, but ciliary function is poorly studied. Recent data suggested that abnormal ciliary beating may contribute to poor MCC in CF patients. However, results are controversial: ciliary beat frequency (CBF) might be either decreased or increased, compared with healthy subjects. We aimed to evaluate if ciliary function may have a role in impaired mucociliary clearance in CF patients and if this ciliary dyskinesia was present in childhood. Ciliated epithelial samples were obtained by nasal brushing from 22 pediatric (0-17 years) and 15 adult CF patients (18-78 years). Beating cilia were recorded using digital high-speed videomicroscopy at 37 °C. Ciliary functional analysis (CFA) was assessed by CBF and the percentage of abnormal ciliary beat pattern (CBP) and was compared with 14 healthy subjects (HS) (22–54 years). There was a significant difference in CBF only between adult CF patients and HS. Abnormal CBP was significantly higher in CF patients, regardless of the age of patients ( Table 1 ). This study suggested that an abnormal ciliary function, present in childhood, might contribute to impaired MCC in CF patients as shown by lower CBF or higher dyskinetic CBP compared with HS. To evaluate if ciliary dyskinesia is primary or secondary to chronic inflammation, CFA should be repeated after air-liquid interface cell culture.

Ciliary beating patterns map onto a low-dimensional behavioural space

Biological systems are robust to perturbations at both the genetic and environmental levels, although these same perturbations can elicit variation in behaviour. The interplay between functional robustness and behavioural variability is exemplified at the organellar level by the beating of cilia and flagella. Cilia are motile despite wide genetic diversity between and within species, differences in intracellular concentrations of ATP and calcium, and considerable environment fluctuations in temperature and viscosity. At the same time, these perturbations result in a variety of spatio-temporal patterns that span a rich behavioural space. To investigate this behavioural space we analysed the dynamics of isolated cilia from the unicellular algae Chlamydomonas reinhardtii under many different environmental and genetic conditions. We found that, despite large changes in beat frequency and amplitude, the space of waveform shapes is low-dimensional in the sense that two features account for 80% of the observed variation. The geometry of this behavioural space accords with the predictions of a simple mechanochemical model in the low-viscosity regime. This allowed us to associate waveform shape variability with changes in only the curvature response coefficients of the dynein motors.