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Biomedical subjects

P Gehr

Publications and source records attributed to P Gehr.

At least 19 recordsLinked to original sources
PubMed1995

Efficiency of airway macrophage recovery by bronchoalveolar lavage in hamsters: a stereological approach.

Macrophages play a central role in the defence of the respiratory tract against deposited particles. In addition to the well-studied alveolar macrophages, airway macrophages have been recognized as an important clearance factor. Bronchoalveolar lavage (BAL) has been used for functional and morphological investigations of macrophages in vitro, assuming that all macrophages are removed with equal probability from the lung surface. Airway macrophages have been found in close contact with the epithelial cells. These macrophages may not be easily removed by lavage, and they might constitute a functionally different macrophage population. We have tested the hypothesis that there exists a population of macrophages in the conducting airways that resists removal by lavage. We lavaged the lungs of four hamsters and fixed the lungs, thereafter, by intravascular perfusion. The number of macrophages in the intrapulmonary conducting airways was estimated with an unbiased stereological technique, the fractionator, and compared to the number of macrophages in the airways of four hamsters whose lungs had not been lavaged prior to fixation. This in situ study revealed that, in hamster lungs, 42% of the airway macrophages were not removed by BAL and that about 5% of all macrophages in the BAL fluid were airway macrophages. Additionally, ultrastructural alterations of the airway epithelium were found. It is concluded that there exists a population of airway macrophages that resists lavage. This is an aspect which has to be considered in studies performed with macrophages obtained by BAL, since they could represent a functionally different macrophage population.

Animals↗
PubMed1994

The effect of particle inhalation on macrophage number and phagocytic activity in the intrapulmonary conducting airways of hamsters.

The number and functions of macrophages in the lungs are crucial factors for prevention and development of lung disease caused by inhaled particles. To examine whether airway macrophages are attracted to the site of particle deposition and what proportion of these macrophages is involved in phagocytosis, aerosols of 6-microns polystyrene particles (PSP) were inhaled by Syrian Golden hamsters under controlled conditions through an inhalation tubule and their lungs were fixed by intravascular perfusion within 20 min (PSP-1, PSP-1a), 40 min (PSP-2), and 24 h (PSP-3) after the beginning of the inhalation. The number and the phagocytic activity of airway macrophages were studied in situ with a fractionator, a stereologic method, on light microscopic sections. No significant increase in macrophage number was detected for the groups PSP-1 and PSP-1a. The increase for group PSP-2 was, however, between 2- and 3-fold, whereas for group PSP-3 the increase was between 1.5- and 2.5-fold with respect to control animals, which had inhaled ambient air through an intubation tubule (C-2) and whose lungs had been fixed after 40 min. There were no significant differences among the four groups with respect to the proportion of airway macrophages that had phagocytized polystyrene microspheres. Twelve to fifteen percent of the macrophages were found to be involved in phagocytosis. In the case of the mean number of particles per phagocytizing macrophage, there was a significant decrease for the PSP-3 group with respect to the pool of the three groups PSP-1, PSP-1a, and PSP-2 taken together. These studies demonstrate (1) that airway macrophages are rapidly recruited to the sites of particle deposition and (2) that only a small proportion of very active macrophages contributes to the clearance of particles, suggesting a great potential of airway macrophages to interact with many more particles than the hamsters were exposed to in this study.

Administration, Inhalation↗
PubMed1993

Surfactant and inhaled particles in the conducting airways: structural, stereological, and biophysical aspects.

We have investigated the displacement into the sol phase of inhaled particles deposited in the intrapulmonary conducting airways. Hamsters inhaled an aerosol of monodisperse polystyrene particles of 6 microns diameter. Their lungs were fixed by intravascular perfusion, and light and electron microscopy was used to study the epithelial coating. The surfactant film at the wall-air interface was investigated by measuring its surface tension. The number of particles retained was determined stereologically. In addition we investigated the displacement of spherical particles in vitro on a DPPC monolayer in a Langmuir-Wilhelmy surface balance and determined the surface tension in vivo in the horse trachea by video bronchoscopy, applying the droplet spreading method. We found that particles deposited onto a surfactant film were pulled into the aqueous subphase, and we concluded that surface forces due to the airway surfactant likely displace deposited particles into the periciliary fluid (sol phase). Comparing lungs fixed immediately after inhalation with lungs fixed 24 hr after inhalation revealed that 86% of the particles retained in the intrapulmonary conducting airways immediately after inhalation had been cleared within 24 hr. One-third of the particles of the lungs fixed immediately after inhalation was phagocytized. The combination of structural and stereological analyses with in vitro and in vivo measurements has led to new insights into the role of airway surfactant with respect to the fate of inhaled particles, which may have important consequences regarding the effects of hazardous particles. These studies may also help to evaluate the deposition pattern and clearance of therapeutic particles, with important implications for their therapeutic use.

Administration, Inhalation↗
PubMed1992

Stereological and functional analysis of liver mitochondria from rats with secondary biliary cirrhosis: impaired mitochondrial metabolism and increased mitochondrial content per hepatocyte.

Mitochondrial and cytosolic functions were studied in vivo and in perfused livers from rats with secondary biliary cirrhosis induced by bile duct ligation for 5 wk and in sham-operated controls. The livers were stereologically analyzed, and mitochondrial and cytosolic functions were related to liver structure. Oxygen consumption by perfused livers expressed per stereologically determined mitochondrial volume was decreased by 49% in bile duct-ligated rats compared with control rats. Glucose production (expressed per mitochondrial volume) was reduced by more than 90% in bile duct ligation, whereas urea production was not affected. Lactate production, a cytosolic function, was increased fivefold in bile duct ligation, and both the lactate/pyruvate and the beta-hydroxybutyrate/aceto-acetate ratios were increased in the liver perfusate of bile duct-ligated rats. In comparison with control rats, the stereologically determined mitochondrial volume fraction per hepatocyte was increased by 28% in bile duct-ligated rats. Activities of mitochondrial enzymes expressed per area of mitochondrial membrane or per mitochondrial volume were either unchanged (ATPase, cytochrome c oxidase and glutamate dehydrogenase) or decreased (monoamine oxidase) in bile duct ligation. Thus in comparison with control rats, mitochondrial metabolism is impaired in perfused livers from bile duct-ligated rats; increased mitochondrial volume per hepatocyte may represent a strategy to maintain hepatic energy metabolism in rats with secondary biliary cirrhosis.

Animals↗
PubMed1992

Hepatic accumulation of lysosomes and defective transcytotic vesicular pathways in cirrhotic rat liver.

To investigate the potential role of lysosomes in cirrhosis, we analyzed the activity of lysosomal enzymes in rats exposed long-term to phenobarbital and carbon tetrachloride. The activity of lysosomal enzymes was markedly increased in the homogenate of cirrhotic livers (e.g., arylsulfatase 9 +/- S.D.2 vs. 16 +/- 6 nmoles.min-1.mg-1 in control rats and cirrhotic rats, respectively; p less than 0.001). The corresponding plasma levels were also increased (7 +/- 1 vs. 12 +/- 3 nmoles.min-1.mg-1; p less than 0.01), whereas biliary excretion was diminished (16 +/- 7 vs. 7 +/- 2 pmol.min-1.gm liver-1; p less than 0.05) in cirrhotic rats. Stereological quantification of lysosomes visualized cytochemically revealed an increase of pericanalicular lysosomes averaging 1.5 +/- 0.4 around a canaliculus in controls and 3.7 +/- 1.0 in cirrhotic rats (p less than 0.01). Because this suggested a defect in the transcellular vesicular pathway, we investigated the biliary excretion of horseradish peroxidase and epidermal growth factor in perfused livers. Bile flow and total horseradish peroxidase excretion were similar in control rats and cirrhotic rats. However, the early peak of biliary horseradish peroxidase excretion--usually taken as evidence of paracellular transport--was increased in cirrhotic rats (13 +/- 7 vs. 57 +/- 22%; p less than 0.01), whereas the second peak--reflecting the transcellular vesicular pathway(s)--was markedly reduced (87 +/- 7 vs. 43 +/- 22%; p less than 0.001). A similar reduction in the biliary excretion of intact epidermal growth factor and of its degradation products was found. These results demonstrate an increased number of lysosomes in hepatocytes of cirrhotic livers; this appears to be the result of accumulation rather than proliferation, in view of the reduced transcellular vesicular movement of different markers into bile.

Animals↗
PubMed1992

Allometric relationships of cell numbers and size in the mammalian lung.

Allometric studies have shown that lung volume, alveolar surface area, and diffusing capacity increase proportionally with body weight across a broad range of mammalian species. Changes in the number of cells and in average cell size and surface areas with increasing body weight have not been defined. We speculated that cell size is determined more by cell function than by species and body weight. To test this hypothesis, nine species ranging in size from shrew (2 to 3 g) to horse (510 kg) were studied. Random sites from the distal alveolar region of each species were analyzed using morphometric techniques. Six to 10 nuclei from each of the major classes of parenchymal lung cells were three-dimensionally reconstructed to determine their average diameter, volume, and surface area. To calculate the cell density, nuclear profiles were counted using electron microscopy. The number of cells per lung increased with body mass and lung volume with a slope of 1.01 (r2 = 0.99). The lung is unique among organs in the diversity and function of individual cell types, such as mechanical, sensory, secretory, transporting, and circulating cells. Excluding the circulatory cells, the lung has greater than 60 different cell types, making it an ideal organ for examining the varieties in cell characteristics across different species. Up to 6-fold differences in size were found between different lung cell types within a single species; however, for cells having secretory functions, such as type II cells, there was no detectable change in cell size with increasing lung surface area or body mass.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

[Particle retention in the intrapulmonary conducting airways in hamsters].

Inhalation and the subsequent deposition, retention and clearance of particles play a pivotal role in pulmonary medicine. It has been widely accepted that inhaled insoluble particles which are deposited in the conducting airways are largely cleared within 24 hours by the mucociliary escalator. Recent reports, however, suggest that this view should be revised. We therefore investigated by light and electron microscopy the number and location of particles retained in the intrapulmonary conducting airways of hamsters as a function of time. The number of retained particles was estimated unbiasedly with a novel stereological method, the fractionator. Seven male Syrian Golden hamsters inhaled an aerosol of 6 microns polystyrene particles under controlled conditions. The lungs of 3 animals were fixed by intravascular perfusion 20 minutes after the inhalation, whereas those of the other 4 animals were fixed 24 hours later. Retained particles were found in all generations of the intrapulmonary conducting airways. They were submerged in the extracellular matrix in close contact with the epithelial cells between the cilia. The particles were coated by a thin layer of osmiophilic material which was interpreted as surfactant. In the lungs fixed immediately after the inhalation, 36% of the retained particles were ingested by macrophages. 24 hours after inhalation we still found 14% of the initially retained particles. 90% of these remaining particles were phagocytized. We conclude that: (1) Surfactant is important for the retention of particles in the intrapulmonary conducting airways of hamsters. (2) There is evidence for the existence of a resident population of very active airway macrophages which play a pivotal role in clearance of the retained particles.(ABSTRACT TRUNCATED AT 250 WORDS)

Air Pollutants↗
PubMed1990

Mitochondrial structure and function in CCl4-induced cirrhosis in the rat.

To investigate whether the impairment of mitochondrial function in cirrhosis is due to a reduction in liver cell mass or whether mitochondrial function is altered specifically, we analyzed mitochondrial volume and surface density of mitochondrial membranes in control and cirrhotic rats by stereological means. Cirrhosis was induced by long-term exposure to phenobarbital and CCl4. Hepatocellular and mitochondrial volumes were reduced to a similar extent, by 39% and 40%, respectively, in cirrhotic animals (p less than 0.01). Thus the fraction of hepatocytes occupied by mitochondria did not differ between the two groups. Both total outer (31 +/- 3 vs. 19 +/- 6 m2; p less than 0.01) and inner (87 +/- 24 vs. 45 +/- 12 m2; p less than 0.01) mitochondrial membranes were significantly reduced. Membrane surface was normal per unit of mitochondrial volume, however, suggesting intact mitochondrial structure. Matrix and outer membrane enzyme activities expressed per compartment did not differ between control and cirrhotic animals. Inner membrane, in contrast, had an increased enzyme content per unit area both for cytochrome oxidase (10.3 +/- 2.9 vs. 13.0 +/- 1.6; p less than 0.05) and ATPase (13.7 +/- 1.4 vs. 21.2 +/- 2.9; p less than 0.01). Basal oxygen consumption measured in the perfused liver in situ was significantly reduced in cirrhotic livers (1.6 +/- 0.1 vs. 1.1 +/- 0.4 mumol/min-1/gm-1) but was unchanged when expressed per square meter of inner membrane. Our results demonstrate that impaired mitochondrial function is mainly due to loss of hepatocellular mass. Increased enzyme activity per unit surface area of inner mitochondrial membrane may be important to maintain mitochondrial function of the cirrhotic liver.

Animals↗
PubMed1990

Surfactant displaces particles toward the epithelium in airways and alveoli.

This study was designed to investigate the early stages of particle deposition on airway and alveolar surfaces. To do this we used morphometric studies of aerosol deposition, in situ measurements of surface tension, and in vitro assays of particle displacement and mathematical modelling. We observed that latex particles, equal or less than 6 microns in diameter deposited in hamster lungs were submerged in the subphase of the alveolar lining layer and became completely coated with an osmiophilic film. Similar results were obtained for particles deposited in the conductive airways which were also covered with a surface active film, having a surface tension of 32 +/- 2 dyn.cm-1. In vitro experiments showed that pulmonary surfactant promotes the displacement of particles from air to the aqueous phase and that the extent of particle immersion depends on the surface tension of the surface active film. The lower the surface tension the greater is the immersion of the particles into the aqueous subphase. Mathematical analysis of the forces acting on a particle deposited on an air-fluid interface show that for small particles (less than 100 microns) the surface tension force is several orders of magnitude greater than forces related to gravity. Thus, even at the relatively high surface tension obtained in the airways (32 +/- 2 dyn.cm-1) particles will still be displaced into the aqueous subphase. Particles in peripheral airways and alveoli likely are below the surfactant film and submerged in the subphase. This may promote clearance by macrophages. In addition, particle displacement into the subphase is likely to increase the contact between the epithelial cell and particle. Toxic or allergenic particles would be available to interact with epithelial cells and this may be important in the pathophysiology of airway disease.

Administration, Inhalation↗
PubMed1990

Assessment of particle retention and clearance in the intrapulmonary conducting airways of hamster lungs with the fractionator.

A modified version of the fractionator was used to estimate the total number of polystyrene microspheres retained in the airways of hamster lungs at two different time points after inhalation. A systematic three-stage subsampling procedure with known sampling fractions was adopted. First, each lung was cut into slices, from which primary disectors were sampled systematically with a known sampling fraction. From each primary disector, smaller sub-disectors were subsampled, and the corresponding sampling fraction was estimated by point counting. Finally, a few particles were counted at the microscopic level in the sub-disectors, and the final estimate of total particle number (which is unbiased irrespective of any tissue deformations) was easily computed as a product of the counted number times the reciprocal of the successive sampling fractions. The error variance of each estimate was assessed from the data using a new estimator. An average of 6% of the deposited particles were retained on the epithelial surface of the intrapulmonary conducting airways shortly after the inhalation, from which at least one-third was already phagocytosed by macrophages. After 24 h, an average of 87% of the particles retained shortly after the inhalation had been cleared. The proportion of particles ingested by macrophages had increased to at least 87% in three out of four animals studied.

Animals↗
PubMed1990

Phagocytic properties and organelle motility of pulmonary macrophages from smokers and nonsmokers estimated in vitro by magnetometric means.

Monolayer cultures of pulmonary macrophages (PM) from 11 smokers (S) and 9 nonsmokers (NS) were incubated with submicrometric ferromagnetic Fe3O4 particles for one hour. After magnetization by an externally applied pulse magnetic field, the aligned Fe3O4 particles, located in the phagosomes and secondary lysosomes of the PM, emanated a remanent magnetic field which decayed with time. The initial field strength, B0, which is proportional to the amount of phagocytosed Fe3O4 particles, was 13.24 nT (SE 0.79 nT) in S and 11.74 nT (SE 1.39 nT) in NS. Ten nT correspond to roughly 4 micrograms Fe3O4. The initial rate of decay of the remanent field (during the first 60 s after magnetization), lambda 0, is proportional to the rate of particle misalignment. Therefore, lambda 0, is hypothesized to be an estimate of organelle motility. It was found to be the same in S and in NS, being 3.14 x 10(-3).s-1 (SE 0.18 x 10(-3).s-1) in S and 3.17 x 10(-3).s-1) (SE 0.22 x 10(-3).s-1) in NS. These results suggest that, in vitro, there is no difference in phagocytic activity and organelle motility in PM from S and NS.

Adult↗
PubMed1989

Histological and clinical parameters of human gingiva following 3 weeks of chemical (chlorhexidine) or mechanical plaque control.

The aim of the present study was to compare stereologically the histopathologic variations following 3 weeks of chemical (chlorhexidine) or mechanical plaque control. 18 students and dental hygienists volunteered for this investigation. After prophylaxis, they performed optimal oral hygiene to reach mean plaque and gingival indices approaching 0. Six of them then performed mechanical plaque control of 3 weeks (control), while the other 12 rinsed 3 times daily with a 0.12% chlorhexidine solution (test). At days 0 and 21, the plaque index (PlI), the gingival index (Gl) and the gingival exudate flow rate (GEFR) were assessed and biopsies were obtained from buccal sites. Point-counting procedures were performed at 2 different levels of magnification on light microscopic sections to estimate the volume fractions of epithelium, infiltrated and non-infiltrated connective tissue, and collagen. The relative numbers of fibroblasts, polymorphonuclear neutrophils, lymphocytes, plasma cells, macrophages and mast cells were estimated by counting the number of nuclear profiles of these cells in a specific connective tissue area adjacent to the apical termination of the junctional epithelium. After 21 days, the PlIs of the test subjects were significantly higher than the PlIs of the controls, but their Gl were similar. At the end of the experimental period, the various volume fractions and %s of cell profiles remained stable with the exception of an increase in the %s of lymphocytes in the test group. This study has shown that, clinically as well as histologically, the daily use of chlorhexidine for a 3-week period is equally efficient as optimal mechanical tooth cleaning in maintaining a healthy gingiva in the buccal sites investigated.

Chlorhexidine↗
PubMed1989

Magnetometric estimation of organelle motion in hamster lung macrophages in vitro: the effects of temperature.

Pulmonary macrophages, containing ingested ferrimagnetic particles, were studied in vitro. Magnetic particles (Fe3O4) suspended in saline were repeatedly instilled intratracheally into the lungs of Syrian golden hamsters. Later the animals were sacrificed and pulmonary macrophages were harvested by repeated lung lavage. After monolayer cultures were established, the particles previously phagocytized by the macrophages were magnetized and aligned by applying an external field. After the external field was removed, the particles produced a remanent magnetic field which gradually decreased due to random misalignment of the particles (relaxation). When relaxation was measured at various incubation temperatures, we found that relaxation slowed as the temperature decreased; we believe that this reflects decreased organelle motion. These results provide direct evidence that relaxation measured previously in intact animals is related to intracellular movements. Magnetometry can be used to quantitate the motion of cell organelles and thus the influence of altered cytoplasmic rheology and cytoskeletal activity.

Animals↗
PubMed1988

Observations on the initial stages of healing following human experimental gingivitis. A clinical and morphometric study.

The aim of the present study was to investigate stereologically the histologic alterations occurring during gingival healing after experimental gingivitis and to compare clinical parameters with histological findings. 8 dental students volunteered for the investigation. After a prophylaxis, they performed optimal oral hygiene to reach mean plaque and gingival indices approaching zero. They then abolished all oral hygiene procedures for a period of 21 days. After this experimental gingivitis phase, they again performed optimal oral hygiene for 8 days to restore gingival health. At days 0, 1, 2, 4 and 8 after experimental gingivitis, the plaque index (PlI), the gingival index (GI) and the gingival exudate flow rate (GEFR) were assessed and their buccal gingiva was biopsied. Point counting procedures were performed at 2 different levels of magnification on light microscopic sections to estimate the volume fractions of epithelium, infiltrated and non-infiltrated connective tissue, and collagen. The relative numbers of fibroblasts, polymorphonuclear neutrophils, lymphocytes, plasma cells and macrophages were estimated by counting the number of profiles of these cells in a specific connective tissue area adjacent to the apical end of the junctional epithelium. A rapid drop in the PlI was noted with increasing time after oral hygiene, followed by a slower decrease in the GI and GEFR scores. The histological picture during the entire experiment was that of an initial gingival lesion. At day 0, no chronic inflammation of the gingiva characterized by a predominance of plasma cells was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1988

Stereological observations on long-term experimental gingivitis in man.

The purpose of the present investigation was to study stereologically the histopathologic changes in the gingiva during 6 months of abolished oral hygiene and to study the development of chronic gingivitis in man. After a thorough prophylaxis procedure, 5 dental students performed optimal oral hygiene under supervision for a period of 3 weeks. At the end of this pre-experimental phase, they were asked to abolish all oral hygiene procedures for 4 (2 individuals) to 6 months (3 individuals). At day 21, and after 2, 3, 4, 5 and 6 months, the gingival exudate flow rate and the gingival index were assessed, and buccal gingival biopsies taken. Semi-thin histologic sections were stained with basic fuchsine and methylene blue. By point counting at 2 different levels of magnification, the volume densities of epithelium, infiltrated (ICT) and non-infiltrated connective tissue, and collagen were estimated. The %s of fibroblasts, PMN's lymphocytes, plasma cells and macrophages were estimated in a predetermined standardized area close to the apical termination of the junctional epithelium. With increasing time, the volume densities of the ICT rose concomitantly with a decrease in the volume densities of the collagen. In spite of great interindividual variations, a slow shift in the proportions of some cell populations was consistently observed. While the fraction of PMN's, lymphocytes and macrophages remained stable, a decrease of fibro-blasts (57 to 39%) and an increase of plasma cells (0.2 to 10%) was observed. This study has, therefore, demonstrated that, in 6 months of plaque accumulation, a chronic gingivitis with a predominance of PMN's and lymphocytes develops.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1988

Spectrin-like proteins in the paraflagellar rod structure of Trypanosoma brucei.

A polyclonal, monospecific rabbit antibody to human erythrocyte spectrins cross-reacted with two sets of proteins (a doublet of 180/200K and a triplet of 67-66-65K; K = 10(3) Mr) in the parasitic protozoon Trypanosoma brucei brucei. Except for the 66K protein, the cross-reacting proteins are localized in the flagellum, on the basis of evidence from cell fractionation and immunofluorescence microscopy. Immunogold labelling and electron micrographs further revealed that the spectrin-like proteins are confined to the paraflagellar rod structure. The spectrin-like proteins with apparent molecular weights of 180 and 200 share homology with spectrin band 1, since V8-protease from Staphylococcus aureus generated similarly sized, antigenic peptides from these proteins. The results indicate homology between the cross-reacting proteins and human red cell spectrin.

Animals↗
PubMed1988

Diacylglycerol-induced shape changes, movements and altered F-actin distribution in human neutrophils.

The study shows that diacylglycerols (DAGs) as physiological activators of protein kinase C induce characteristic shape changes in human neutrophil granulocytes. In contrast to chemotactic peptides, which can induce front-tail polarity characterized by a contracted tail and an expanding front, DAGs elicit the formation of non-polar cells with surface projections. These cells exhibit a distinct type of motility characterized by vigorous and continuous shape changes without front-tail polarity and without the unidirectional movement and cytoplasmic streaming seen in polarized cells. In neutrophils exposed to DAGs, F-actin is shifted to the cell periphery and mainly into the surface projections of activated cells. DAGs induce the formation of large intracellular vacuoles in neutrophils producing surface projections, and these vacuoles persist after the cells have reacquired a spherical shape. Combined stimulation of human neutrophils with DAG and fNLPNTL results in a suppression of peptide-induced polarity and the formation of non-polar motile cells resembling those stimulated with DAG alone. These results suggest that the diacylglycerol-protein kinase C pathway may be instrumental in transducing or modulating signals to both the locomotor apparatus and the exocytotic and/or pinocytic system of the cell. Neutrophil stimulation with DAGs thus represents a useful model with which to study further the hypothesis that distinct types of neutrophil shapes and movements are preferentially associated with distinct functions and to characterize signalling pathways.

Actins↗