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Effects of prostaglandins D2 and E2 on ciliary beat frequency of human upper respiratory cilia in vitro.

Diminished mucociliary transport can occur in a type-I (Ig-E mediated) allergic reaction. We determined the effects of the allergy mediators prostaglandin D2 (PGD2) and prostaglandin E2 (PGE2) on the ciliary beat frequency (CBF) of human upper respiratory cilia in vitro. Human adenoid tissue was used as the source for ciliated epithelium. CBF was measured by a computerized photo-electric method. PGD2 (10(-8)-10(-5) M, n = 7) showed no statistically significant effect on CBF. PGE2 (10(-9)-10(-6) M, n = 10) caused a significant dose-dependent stimulation, with a maximum of 37% (ANOVA, p < 0.001). Thus prostaglandins D2 and E2 do not exert a direct negative influence on ciliary activity, which could account for a decrease in mucociliary transport. The stimulating effect of PGE2 may be relevant in promoting mucociliary clearance in vivo.

Adenoids↗

Intracytoplasmic lumina with and without cilia in both normal and pathologically altered nasal mucosa.

Two different types of intracytoplasmic lumina have been observed electron microscopically in pseudostratified and metaplastic human nasal mucosa: one type contains microvilli and the other contains both microvilli and cilia. The latter type was seen only in pseudostratified columnar epithelium, while the former was also observed in different stages of metaplasia. From these observations we conclude that the type of intracytoplasmic lumina is determined by the differential potential of the cells in which they occur. Serial sectioning demonstrated that these structures sometimes communicate with the extracellular space. The distinctly different ultrastructure of microvilli lining the intracytoplasmic lumina and the foldlike extensions of the cell membrane favors an intracellular genesis rather than a formation by invagination of the cell membrane. It is suggested that intracytoplasmic lumina occur as a consequence of dysregulation in the process of epithelial maturation.

Cilia↗

Atypical cilia in hyperplastic, metaplastic, and dysplastic human bronchial mucosa.

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.

Adult↗

Direct measurement of the velocity of the metachronal wave in beating cilia.

Recently a computerized electro-optical method was developed which enables one to simultaneously measure the frequency and the wavelength of the metachronal waves in beating cilia. The method is based on measurement of scattered light from two areas at a given distance apart. The distance between measured areas can be varied from zero to hundreds of microns. The relative ease of the measurement and data analysis of this method enable one to create large statistical ensembles in order to obtain reliable averages. In this work we show that in addition to the previously mentioned parameters this system can measure directly the velocity of the metachronal wave. It was found that the average velocity in the tissue culture from frog's palate epithelium at room temperature is approximately 270 micron/sec, about twice the average particle velocity at the frog's palate.

Animals↗

Characterization of metachronal wave in beating cilia: distribution of phases in space.

We have examined phase differences between beating cilia measured in different directions on the surface. The measurements were performed on tissue cultures taken from frog palate epithelium. This experiment was based on a method developed earlier, which measures simultaneously scattered light from two different and relatively small areas. Moreover, the distance between the two small areas can be accurately defined and varied. Rotating the stage around the optical axis permits us to repat these measurements at different space angles. It was found that: a) the phase gradient is a periodic function of the angle of rotation; b) the periodicity can be described quantitatively by a cosine function; c) good agreement exists between Machemer's findings in protozoa and our results, despite the differences between the two systems and techniques used.

Animals↗

Cilia-associated respiratory (CAR) bacillus infection in adult red deer, chamois, and roe deer.

Cilia-associated respiratory (CAR) bacillus is an unclassified bacterium that colonizes the ciliated epithelium of airways in laboratory rats, laboratory mice, and laboratory and conventionally reared rabbits, cattle, goats, and pigs. Data on the prevalence of CAR bacillus infection in wild animals are lacking. The present study demonstrated the occurrence of the organism in wild red deer (Cervus elaphus hippelaphus), chamois (Rupicapra rupicapra), and roe deer (Capreolus capreolus) from the Val Fontana in northern Italy. Prevalence ranged from 26% for red deer to 56% for chamois, with a statistically significant negative correlation between CAR bacilli infection and the presence of lymphoid follicles.

Animals↗

Antigenic analyses of cilia-associated respiratory (CAR) bacillus isolates by use of monoclonal antibodies.

Mouse monoclonal antibodies (MAbs) developed to a rat isolate (R-3) of cilia-associated respiratory (CAR) bacillus were used to assess antigenic relationships among three rat and five rabbit CAR bacillus isolates. Evaluation of MAbs by enzyme-linked immunosorbent assays (ELISAs) indicated that 87 of 241 hybridomas secreted CAR bacillus-reactive antibodies that could be grouped into four major groups. Group-I MAbs reacted with epitopes expressed by all CAR bacillus isolates and at least two or more nonrelated species of bacteria. Group-II, -III, and -IV MAbs reacted with only one or more of the rat CAR bacillus isolates; no MAbs reacted only with rat and rabbit CAR bacillus isolates. Western blot analyses indicated that 41-, 50-, and 105-kDa peptides of rat CAR bacillus isolates expressed rat CAR bacillus group- and isolate-specific epitopes. Hyperimmune anti-CAR bacillus antiserum and serum specimens from a CAR bacillus histologically positive mouse and rat also reacted with the 41-, 50-, and 105-kDa peptides. Sera from CAR bacillus histologically negative rats did not react with these peptides. These results suggest that the 41-, 50-, and 105-kDa peptides may represent suitable antigens for development of a specific ELISA for detection of rodent CAR bacillus infections. Furthermore, these data indicate that use of crude CAR bacillus preparations for either rat or rabbit CAR bacillus ELISAs is inappropriate.

Animals↗

Function and ultrastructure of cilia in primary ciliary dyskinesia.

Primary ciliary dyskinesia (PCD) is a heterogeneous disease with impaired mucociliary transport leading to upper and lower respiratory disorders, hearing impairment and male infertility. Primary ciliary dyskinesia can only be diagnosed by clinical features together with functional and structural analysis of the cilia. To prevent bronchiectasis with marked reduced quality of life, early diagnosis is essential. For this purpose we compared our experience over 10 years with the literature. Our concept consists of a thorough interdisciplinary examination of the patient to rule out other underlying pathologies such as allergy, cystic fibrosis, immune deficiencies, and alpha-1-antitrypsin deficiency on the basis of their clinical features. Thereafter, mucosal biopsies from 27 patients were investigated. In 10 patients (37%) primary ciliary dyskinesia was diagnosed with the help of functional and ultrastructural analysis. 9 patients displayed no or impaired ciliary motility and a high percentage showed ultrastructural defects such as dynein arm deficiency, radial spoke defects and translocation of peripheral microtubular doublets with absent central microtubules. We suggest that investigation of mucosal biopsies for primary ciliary dyskinesia diagnosis without clinical preselection is indicated in patients with "situs inversus" suffering from chronic and/or recurrent airway infections, in patients with neonatal respiratory distress syndrome of "unknown" cause (i.e. after ruling out the other well known causes) with "situs inversus".

Adult↗

Cilia-Associated Respiratory (CAR) bacillus infection in veal calves and adult cattle.

Cilia-Associated Respiratory (CAR) bacillus is a filamentous bacterium that colonizes the ciliated epithelium of the respiratory tract of many animal species and that has been associated with chronic inflammatory lesions in naturally and experimentally infected rats, mice and rabbits. In the present study, the prevalence of CAR bacillus infection and histological lesions of the trachea in veal calf and adult cattle were investigated. Forty five healthy veal calves and 45 adult cattle, raised in 18 different herds were selected at slaughter. From each animal, a tracheal sample was processed for histology, stain-ed with the Warthin-Starry method to evaluate the presence of CAR bacillus, and with haematoxylin and eosin to evaluate the presence of inflammatory lesions. CAR bacillus was identified in 17 veal calves (37.7%) and in 7 adult cattle (15.5%). Inflammatory lesions were found in 42 veal calves (93.3%) and in 41 adult cattle (91.1%). Statistical analysis pointed out a significant correlation between the presence and number of CAR bacilli and the presence and number of lymphoid follicles (P = 0.0071) and the presence and severity of neutrophilic infiltrates (P = 0.0428). These results indicate that CAR bacillus infection is common in cattle and is correlated with tracheal inflammatory lesions.

Animals↗

Application of the cilia dyskinesis test in the diagnosis of cystic fibrosis in an adolescent.

In certain cases of obscure chronic lung disease, the possibility of a genetic disorder, such as cystic fibrosis of the pancreas, should be considered. When the genetic defect fails to cause pancreatic involvement, the partial expression of cystic fibrosis is encountered. Elevation of sweat electrolyte levels is not sufficient to confirm the diagnosis of cystic fibrosis in adults since normal levels vary widely in both adolescents and adults. Other tests should, therefore, be utilized. The cilia dyskinesis test is positive in both homozygous and heterozygous patients with cystic fibrosis. Utilization of this technique in a family study confirmed the diagnosis of partial expression of cystic fibrosis in an adolescent female.

Acid-Base Equilibrium↗

[Morphologic changes in photoreceptor connecting cilia in experimental phototoxic retinopathy].

PURPOSE: It has been suggested that structural alterations of the photoreceptor connecting cilium are a primary defects leading to photoreceptor degeneration in some forms of inherited retinal degenerations (5). In a series of 17 eyes with retinitis pigmentosa (RP) and with various genetic forms of RP, examined by electron microscopy, no structural abnormalities were found in the arrangement of the ciliary microtubules. However, a 10% reduction in the ciliary diameter was recorded in RP photoreceptors (12). The question arose: is thinning of the cilium a primary defect associated with RP, or a secondary abnormality related to degenerative processes in photoreceptors irrespective of the cause? The aim of this study was to examine the photoreceptor connecting cilia in the early stages of experimental light induced retinal degeneration in the rat, by conducting a structural and morphometric survey of the relevant electron-micrographs. In addition, the effects of various fixation techniques on the ciliary structure were compared. MATERIAL AND METHOD: Analysis of 124 transmission electron micrographs of 9 controls and 155 transmission electron micrographs of 55 light damaged animals was conducted. For the effects of fixation on morphometry 72 transmission electron micrographs from an additional 4 controls (43 negatives) and 8 light damaged animals (29 negatives) were examined. Light damage was induced by exposure to 1000 lux of white light for 120 minutes. Retinal samples were fixed either in 2.5% glutaraldehyde or by high pressure freezing followed by freeze-substitution. RESULTS: This study showed that one of the early morphological alterations occurring in rat photoreceptors damaged by light is a reduction of ciliary diameter of approximately 10%. It was not associated with any apparent ultrastructural changes in the axoneme. It was also found that the degree of ciliary shrinkage largely depends on the fixation technique used. Cryo-fixation followed by freeze substitution shows more shrinkage than chemical fixation by immersion in glutaraldehyde. CONCLUSIONS: It is suggested, that reduction in photoreceptor cilium diameter is a secondary and non-specific change. It is not a unique phenomenon, observed not only in human photoreceptors, which are undergoing degeneration in RP. It can be induced in otherwise healthy rat retina, in which photoreceptor degeneration was caused by exposure to toxic levels of light.

Animals↗

[Mathematical modeling of movement of cilia in olfactory cells].

A mathematical model of the movement of olfactory cilia in different conditions was constructed. The realization of the model includes the development of a mechanical mathematical rheological model of the behavior of a continuous deformable medium, the development of the method of solution adapted to this problem, and obtaining a numerical solution, which takes into account different starting data. The mathematical modeling of the dynamic behavior of a deformable medium was performed using a system of equations for the dynamics of the deformable medium and the solution of the corresponding nonstationary system of equations in partial derivatives.

Cell Movement↗

[Effects of ephedrine on human nasal cilia movement measured with high-speed digital microscopy].

OBJECTIVE: To investigate the effects of ephedrine on human nasal cilia movement. METHODS: Ciliary beat frequency (CBF) of cultured human nasal epithelial cells was measured by high-speed digital microscopy in HBSS and ephedrine solution of different concentrations in 10 minutes. RESULTS: CBF of cultured nasal epithelial cells exposed to HBSS showed no significant changes in 10 minutes. However, in 2.5 g/L , 5 g/L, 10 g/L and 20 g/L ephedrine solution, CBF increased significantly in 1-2 minutes and reached the apex, then it decreased gradually, at the 10th minute. CBF of the samples exposed to 2.5 g/L and 5 g/L ephedrine solution were slower than those in HBSS, but no significant changes were found. However, in 10 g/L and 20 g/L ephedrine solution, CBF decreased significantly when compared with samples in sHBSS. With the concentrations from 2.5 g/L to 20 g/L ephedrine, the increment was independent on the concentration, the inhibitory effect was dependent on the concentration. CONCLUSIONS: In initial time, 2. 5 g/L-20 g/L ephedrine stimulated CBF, then 10 g/L-20 g/L ephedrine inhibited CBF. The stimulation of 2.5 g/L and 5 g/L ephedrine on CBF was longer than that of 10 g/L and 20 g/L ephedrine. 5 g/L ephedrine had maximum stimulatory effect without obvious inhibitory effect on cultured human nasal CBF.

Cells, Cultured↗

[Changes in the motor apparatus of the bronchial cilia. Comparative study of two types of myopathies: nemaline myopathy and the Werdnig-Hoffmann syndrome].

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.

Bronchi↗

Structure of the alpha-, beta-, and gamma-heavy chains of 22 S outer arm dynein obtained from Tetrahymena cilia.

Here we document the UV-induced, vanadate-dependent cleavage of the alpha-, beta-, and gamma-heavy chains of 22 S outer arm dynein obtained from Tetrahymena cilia. All three polypeptides have a single site of photocleavage in the presence of Mg2+, ATP, and vanadate (termed V1 cleavage). The alpha-chain yields complementary fragments with masses of 232 and 185 kDa, the beta-chain has complementary fragments with masses of 225 and 195 kDa, and the gamma-chain has complementary fragments with masses of 242 and 161 kDa. In the absence of ATP, only the beta-chain undergoes V1 cleavage. All three polypeptides have one single site of V2 cleavage, which are unaffected by the presence of nucleotide and only require the presence of Mn2+ and vanadate. V2 cleavage always occurs on the larger V1 fragments and is separated from the V1 site by 52, 48, and 57 kDa for the alpha-, beta-, and gamma-heavy chains, respectively. We have also found a third type of UV-induced vanadate-dependent cleavage which we have termed VMT cleavage. VMT cleavage occurs when dynein is bound to microtubules in an ATP-sensitive manner under V1 solution conditions that should only support cleavage of the beta-chain (i.e. vanadate, Mg2+, and absence of ATP). Under these conditions V1 cleavage of the beta-chain and V2 cleavage of all three chains occur. This is the first documented evidence of V2 cleavage occurring under V1 solution conditions and implies a change in dynein structure when it binds to a microtubule. Using a combination of polyclonal and monoclonal antibodies, we have been able to construct linear polypeptide maps of all three heavy chains. Their relationship to the polypeptide maps previously obtained for heavy chains obtained from the dynein of Chlamydomonas and sea urchin axonemes is discussed.

Adenosine Triphosphatases↗

Primary cilia in normal human neocortical neurons.

Human neocortical neurons from the frontotemporal region were examined by serial ultrathin sectioning with the intention to establish the morphology and frequency of primary cilia (with a 9 + 0 tubular structure) in these cells. Each cell examined thoroughly showed one single cilium with a characteristic position and structure. The constant presence of these organelles may be a hint at an important function.

Cerebral Cortex↗

Kartagener syndrome with abnormalities of cilia.

Kartagener syndrome of sinusitis, bronchiectasis and situs inversus has been so called since his classical description in 1933. This report presents two further cases, a brother and sister. They were extensively studied and the findings are reported. A unique finding was that in both patients, both in the maxillary sinuses and in the bronchi, abnormal cilia were discovered on electron microscopy. After a search of the literature the authors conclude that this finding is probably congenital rather than acquired and may explain the development of the sinusitis and bronchiectasis.

Adolescent↗