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Biomedical subjects

M Rautiainen

Publications and source records attributed to M Rautiainen.

36 records · Page 2Linked to original sources
PubMed1994

Ciliary motility and surface morphology of cultured human respiratory epithelial cells during ciliogenesis.

We used human respiratory epithelial cells in floating culture to study the motility and surface morphology of the cilia during ciliogenesis. Ciliary motility and surface morphology was examined by a high-speed video system and electron microscopy, respectively. On day 8 of floating culture, but rarely before, cells with numerous moving short cilia with mean ciliary beat frequency (CBF) of 17.8 +/- 2.5 Hz (mean +/- SD) were seen. The mean CBF was highest on day 10 (19.6 +/- 3.9 Hz) and reached 17.4 +/- 2.5 Hz on day 14. The ciliary beat amplitude (CBA) was recognized on day 8, abruptly increased on day 9, then continued with aging of the culture. The intracellular coordination of ciliary beats was poor on day 8 with random beat direction but improved up to day 12 when the intracellular uniform beat direction was established. However, intercellular coordination remained absent even on day 14. Results suggest that immature short cilia have a rigid beat pattern. With the elongation of the resulting whip-like movement, the ciliary beat was gradually synchronized. Although a ciliary elongation seems to be required for making intracellular coordination, other factors are involved in intercellular coordination. Also, a high CBF does not always indicate the activity of mature cilia.

Cell Membrane↗
PubMed1994

Effect of exogenous ATP and physical stimulation on ciliary function impaired by bacterial endotoxin.

The effects of exogenous ATP and physical stimulation on ciliary beat depressed by lipopolysaccharide from Pseudomonas aeruginosa were studied in cell culture. Both the ciliary beat frequency (CBF) and the amplitude (CBA) of human respiratory cells in monolayer cell cultures were studied by using a differential interference microscope equipped with a high speed video. Both the ATP and the physiol stimulation stimulated temporarily the depressed CBF and CBA. In both groups the CBF and CBA increased in 1 min to the initial level and then gradually decreased to the level before the stimulation. The duration of the ATP stimulation on the CBF (10.1 +/- 1.8 min) and CBA (10.1 +/- 2.3 min) were significantly better than the duration of physical stimulation on CBF (8.0 +/- 1.8 min) and CBA (6.8 +/- 1.6 min). However, the areas under the CBF/CBA curves (from the beginning of stimulation until the initial level of CBF or CBA was reached again) did not differ significantly. After the removal of the bacterial toxin the CBF was restored to the initial level. In the present study the adenosine receptor antagonist could not prevent the ciliostimulative effect of exogenous ATP combined with the physical stimulation. Both exogenous ATP and physical stimulation have a clear but temporary stimulative effect on the ciliary beat depressed by bacterial toxin, ATP being slightly more effective. It seems that the effects of ATP are not mediated by adenosine receptors.

Adenosine Triphosphate↗
PubMed1993

Degeneration of human respiratory cell ciliary beat in monolayer cell cultures.

The degeneration of ciliary beat of human respiratory cells was studied in monolayer cell cultures by using a differential interference microscope equipped with a high speed video system. This method for studying ciliary beat in cell cultures on collagen-coated cover glasses is quite advantageous, because it allows for detailed study of all parts of ciliary function and not just ciliary beat frequency (CBF). In the present study both CBF and ciliary beat amplitude (CBA) were found to decrease continuously from the 1st day after plating but the wave form of ciliary beat did not change. Cultures with high cell density provided better preservation of normal ciliary beat for a longer period. In contrast, ciliary beat degenerated quickly in cultures with low cell density. CBF and CBA in cell cultures less than 5 days after plating were always high, supporting use of these cultures for studies of normal ciliary motility.

Cell Movement↗
PubMed1993

Effect of exogenous ATP on ciliary beat of human ciliated cells studied with differential interference microscope equipped with high speed video.

To study the detailed effects of exogenous adenosine triphosphate (ATP) on ciliary function, we used the differential interference microscope equipped with high speed video and evaluated ciliated cells from the human sinus mucosa in monolayer culture. With this system it was possible to evaluate all parts of ciliary motility with a minimum of interference from the mucous membrane, secretory cells and the autonomic nervous system. The best direct ciliostimulative effect of exogenous ATP on ciliary beat frequency (CBF) and ciliary beat amplitude (CBA) was observed at concentrations of ATP ranging from 10(-5) M to 10(-3) M. Exogenous ATP appeared to normalize the slightly damaged ciliary motility in groups with an initial CBF of 14 Hz or higher. When the CBF was less than 14 Hz, ATP produced an increase in CBF greater than 40%, an increase in CBA greater than 30%, but these did not reach the normal level in 5 min. The biggest increases: 59.9% in CBF and 40.7% in CBA were seen in the group with an initial CBF less than 9 Hz. In the cells with a low initial CBF and unsynchronized motility exogenous ATP increased the synchrony of ciliary movement together with an increase in CBF and CBA.

Adenosine Triphosphate↗
PubMed1992

Ultrastructural changes in human nasal cilia caused by the common cold and recovery of ciliated epithelium.

Changes in the ultrastructure of human respiratory cilia caused by the common cold were studied in 12 patients. The nasal mucosa was studied three times: on the first or second day after the beginning of symptoms, and 1 week and 3 weeks after the first biopsy. The damage was most severe at 1 week. The most remarkable finding was the loss of cilia and ciliated cells. However, the ultrastructure was usually normal, without any increase in tubular anomalies, as compared with the normal material of the previous reports. Three weeks after the beginning of the disease the number of cilia and ciliated cells had increased to nearly normal. However, as a sign of regeneration, immature short cilia (0.7 to 2.5 microns in length) were often seen. The ciliary orientation was uniform, dynein arms were normal, and there was no increase in the number of tubular anomalies. The results suggest that the impaired mucociliary function during viral infections is due to the loss of cilia and ciliated cells, rather than to ultrastructural anomalies in the cilia. The development of tubular anomalies and random ciliary orientation may require more extensive exposure to factors affecting ciliary function.

Biopsy↗
PubMed1992

Ciliary beat of cultured human respiratory cells studied with differential interference microscope and high speed video system.

The ciliary beating of upper respiratory tract cells cultured on cover glasses was studied by using differential interference microscope equipped with high speed video. By culturing the cells on collagen coated cover glasses, objectives with higher magnifications could be used. With this system we could evaluate not only ciliary beat frequency, but also amplitude, wave form, orientation and synchrony of ciliary beating. Also some structural anomalies such as compound cilia and tide cilia bundles could be recognized. Ciliary beat frequency measured from 1,026 ciliated cells was 20.6 +/- 4.7 Hz (mean +/- SD). The orientation of ciliary beat directions was random and the mean standard deviation for measured angles was 73.0 degrees +/- 28.9 degrees (mean SD +/- SD). When the ciliary beat frequency was 20 Hz, the time used for effective phase was 0.022 +/- 0.002 s (mean +/- SD), and 0.028 +/- 0.004 s (mean +/- SD) for the recovery phase of beat. This system is advantageous for studying ciliary function because all parts can be studied simultaneously with higher magnification, and the effects of chemical physical mediators can be studied without disturbing effects of the autonomic nervous system or secretory cells. Also, the same cells could be observed before and after challenge with test medication and thus evaluated more accurately.

Cilia↗
PubMed1992

Nasal sensitization of dairy farmers to bovine epithelial and urinary antigens.

Nineteen dairy farmers with nasal symptoms associated with working in cowhouses participated in the study. Nasal challenge with bovine epithelial antigen (BEA) and bovine urinary antigen (BUA) was made before and after the indoor feeding season. Nasal challenge made before the indoor feeding season with BEA was positive in five patients and three of them showed positive reaction in nasal challenge also with BUA. After the indoor feeding season the results in nasal challenge with BEA were approximately equal to BEA and four of them showed positive response in nasal challenge to BUA. However, we did not find any significant increase in sensitivity in nasal challenge to BEA or BUA after the indoor feeding season. In addition to these patients, two patients who were excluded from nasal challenge before the indoor feeding season showed positive results in nasal challenge after the indoor feeding season with both BEA and BUA. Our results suggest that BUA in addition to BEA may have significance to nasal symptoms.

Adult↗
PubMed1991

Short nasal respiratory cilia and impaired mucociliary function.

A 48-year-old man was examined because of anosmia. He was otherwise healthy except for mild arterial hypertension. He had stopped smoking 20 years ago. Previously he had been exposed to sulfuric acid gases for 3 years in his work. Clinical examination revealed no findings to explain his anosmia. The patient was fertile, indicating normal sperm/cilia motility. Nasal mucociliary function was examined by radioactive tracer and found to be markedly and constantly impaired. Ciliary ultrastructure in cross-sections was normal. However, in longitudinal sections the length of the cilia varied from 0.6 microns to 3.9 microns. The mean length of the cilia from the cell membrane to the tip was 2.5 +/- 0.9 microns, in contrast to normal ciliary length of 5-7 microns. These findings represent a new structural defect among the various known ciliary abnormalities.

Cilia↗
PubMed1990

Ciliary orientation in the "immotile cilia" syndrome.

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

Biopsy↗
PubMed1990

Comparative analysis of bovine extracts by immunoblotting and ELISA inhibition. Rabbit and human responses against cow epithelial and urine antigens.

Bovine epithelial and urine antigen extracts were compared using enzyme-linked immunosorbent (ELISA) inhibition assay and sodium-dodecyl-sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with immunoblotting. According to ELISA inhibition results, the two extracts share about 2% of their antigenic components. Urine-derived antigens seem to be antigenically closer to serum proteins than epithelial antigens, as determined by rabbit immune sera. The IgG responses of allergic farmers against epithelial antigens were directed mainly against a protein of 20 kD, while non-allergic farmers had only very weak reactions. Generally, the IgG response against urinary antigens seemed to be more heterogeneous than against epithelial antigens. Almost all cow-allergic farmers reacted with a band at 20 kD in IgE immunoblotting against urinary and epithelial antigen while all non-allergic farmers showed negative results.

Animals↗
PubMed1988

Herpes simplex virus detection by macroscopic reading after overnight incubation and immunoperoxidase staining.

Human diploid foreskin fibroblast cells grown in 24-well plates were inoculated with clinical specimens by centrifugation at 1,000 X g for 45 min. Cultures were incubated at 37 degrees C overnight, fixed, and stained with peroxidase-labeled monoclonal antibodies against herpes simplex virus types 1 and 2. Stained plaques of infected cells were large enough to be detected with the naked eye, and microscopic examination did not reveal any further positive specimens. The method was compared with standard isolation in human fibroblasts grown in shell vials and inoculated by centrifugation at 4,000 X g, observed microscopically for the occurrence of typical cytopathogenic effect three times a week for 10 days, and then typed by enzyme immunoassay. Of the 289 specimens tested, 105 were positive and 174 were negative by both methods. Six specimens were positive by standard isolation only, two of them containing varicella-zoster virus, and two specimens were stored frozen before being tested by immunoperoxidase staining. Two specimens found negative by standard isolation were positive by immunoperoxidase staining. For two specimens negative by immunoperoxidase staining, the standard isolation cultures were lost due to microbial contamination. Forty-two specimens found positive by standard isolation were clearly positive when stained only 8 h after inoculation. By standard isolation, positive results were reported on the average 3 to 4 days after inoculation, whereas by immunoperoxidase staining the result was available within less than 24 h. Immunoperoxidase staining of infected cells is a sensitive method for rapid laboratory diagnosis of herpes simplex virus infections, and 24-well plates are convenient for the handling of a large number of specimens.

Antibodies, Monoclonal↗
PubMed1987

Artifacts in ultrastructure of respiratory cilia caused by various fixation procedures and different types of handling.

The ciliary ultrastructure with different fixatives and buffer combinations and artifacts caused by handling the specimens was studied in samples from respiratory mucosa. After different standard fixatives the ciliary ultrastructure did not vary much. Glutaraldehyde in cacodylate gave the best result; the contrast was good and the cell membrane did not show undulation and so many blebs as with other fixatives. With old 4% formaldehyde + 1% glutaraldehyde inphosphate the cytoplasm was darker and the contrast was not as good as with a fresh fixative. Mechanical damage caused rupture of the cell membrane and sometimes changes in axonemal architecture. These findings are similar to those often seen in human samples. The axonemal architecture was always intact if the cell membrane was intact.

Animals↗
PubMed1986

A method for measuring the orientation ("beat direction") of respiratory cilia.

We have created a method for measuring the orientation of cilia in the respiratory epithelium. Ciliary orientation is the direction perpendicular to the plane defined by the central tubules of the cilia and is an estimate of ciliary beat direction. Ciliary orientation can be estimated by measuring the angle between the plane defined by the central tubules and a reference line. The standard deviation of these measurements describes the variation present in the beat directions of the cilia. The reference line must be so chosen that the majority of measurements falls at about the middle of the 0 degree-180 degrees range. We tested measurements by using both a glass angle measure and a semiautomatic image analyzer (IBAS I). The latter approach was faster and more reproducible. We made our measurements of normal tissues on samples obtained from two healthy adult non-smokers. Measurements were made in four areas of each sample, with 59-110 cilia in each. The differences between the maximum and minimum angles of the ciliary orientation in the same area varied from 167.9 degrees to 85.4 degrees from the reference line. The standard deviation varied from 18.0 degrees to 35.4 degrees and we consider this to be a normal variation in ciliary orientation. Of the cilia, 57% were within a standard deviation of 20 degrees.

Adult↗
PubMed1985

Exercise capacity in subjects with high oxygen affinity.

Exercise capacity and hemodynamic parameters were measured in ten patients with a previously unpublished variant of hemoglobin (Hb-Linköping) and in ten age- and sex-matched controls. Bicycle ergometer test was almost maximal and the indices of working capacity and cardiac tolerance were similar in patients and controls. The hemoglobin dissociation curve was shifted to the left at rest and after exercise the shift to right was half of the corresponding shift in controls. Lactate accumulation during exercise was similar in patients and controls, also the elimination rate seems identical, but requires further studies. Hydrogen ion production in the patients during exercise was more marked than in the controls. If this is a sign of altered energy metabolism remains to be ascertained in future studies. In acute short-term exercise the patients with abnormally high hemoglobin oxygen affinity seem to have similar tolerance and cardiovascular load. Aberrant metabolism and adaptation to relative hypoxia in spite of erythrocytosis may have long-term effects that require long-term follow-up of these patients.

Adult↗
PubMed1984

Ultrastructure of human respiratory cilia: a study based on serial sections.

Ultrastructure of human respiratory cilia was studied in ultrathin serial sections and in random fields of sections. The apex of the cilium had dense gray material under the plasma membrane, where the "central" pair of tubules (often slightly eccentrically placed) and the single peripheral tubules ended. Thereafter the cilium thickened gradually about 1 micron below the apex and after that remained reasonably constant. At about 0.5 micron below the apex was a transformation zone (0.1 micron long), where both double and single peripheral tubules were seen. Dynein arms were not seen in the transformation zone but occurred approximately 0.1 micron below it. In a cilium, one peripheral double tubule was absent in the transformation zone but was seen about 0.2 micron below. This resulted in an artificial 8 + 2 pattern in a few sections. In cross sections at levels with peripheral double tubules the mean of the minor axes of the cilia was 0.198 +/- 0.009 micron (+/- SD) calculated from the serial sections of one patient. In 20 patients the variation was greater and the mean was higher: 0.210 +/- 0.042 micron (+/- SD).

Adult↗
PubMedDate not supplied

Nasal airway volumetric measurement using segmented HRCT images and acoustic rhinometry.

Semiautomatic segmentation methods using High Resolution Computed Tomography (HRCT) or Magnetic Resonance Imaging give accurate and reproducible volumetric measurements in various intracranial diseases. In this prospective study, for the first time in literature, with the help of a new semiautomatic segmentation technique and coronal HRCT, we correlated the volumes and cross-sectional areas of the nasal cavity with those obtained by clinical acoustic rhinometry in 14 patients with chronic sinusitis. The measurements obtained by both techniques showed statistically significant correlations between volumes in the anterior and middle parts, but statistically poor correlations between the volumes in the posterior part of the nasal cavity. Coronal HRCT and our new microcomputer applicable semiautomatic segmentation software proved compatible with daily clinical practice. Based on the promising results of our study, we recommend the use of this technique in the validation studies of acoustic rhinometry and in complicated cases as a complementary examination in the evaluation of nasal cavity.

Acoustics↗