Biorheology - Volume 27, issue 3-4

Authors: Mège, Jean-Louis | Capo, Christian | André, Pascale | Benoliel, Anne-Marie | Bongrand, Pierre

Article Type: Research Article

Abstract: The interaction between granulocytes and endothelial walls may be influenced by the blood flow. This possibility was investigated by studying the influence of fluid flow on the adhesion and detachment of 51 Cr-labeled rat granulocytes interacting with protein-coated glass surfaces. It is concluded that: i) Adhesion is markedly decreased when the wall shear rate becomes higher than about 20 s−1 . ii) Pretreating glass with concanavalin A or polylysine significantly decreased adhesion, whereas fibronectin had little effect on binding. iii) Very high flow rates (about one thousandfold higher than those compatible with bond formation) were required to provoke …substantial detachment of substrate-bound cells. iv) Coating glass with laminin or polylysine decreased binding strength whereas fibronectin or concanavalin A did not substantially influence this parameter. v) Exposing granulocytes to phorbol myristate acetate might increase the cell ability to form strong adhesions, whereas labile adhesion was unaffected or even decreased by this treatment. Adhesion is markedly decreased when the wall shear rate becomes higher than about 20 s−1 . Pretreating glass with concanavalin A or polylysine significantly decreased adhesion, whereas fibronectin had little effect on binding. Very high flow rates (about one thousandfold higher than those compatible with bond formation) were required to provoke substantial detachment of substrate-bound cells. Coating glass with laminin or polylysine decreased binding strength whereas fibronectin or concanavalin A did not substantially influence this parameter. Exposing granulocytes to phorbol myristate acetate might increase the cell ability to form strong adhesions, whereas labile adhesion was unaffected or even decreased by this treatment. Show more

Keywords: Adhesion, Granulocyte, Flow

DOI: 10.3233/BIR-1990-273-420

Citation: Biorheology, vol. 27, no. 3-4, pp. 433-444, 1990

Authors: Freyburger, Geneviève | Belloc, Francis | Boisseau, Michel R.

Article Type: Research Article

Abstract: Pentoxifylline (PTX) has been reported to enhance the early accumulation of neutrophils at the site of Staphylococcus aureus subcutaneous infection in mice (1) and to stimulate in vitro PMN chemotaxis, particularly under dense agarose (2). Among the biochemical events contributing to chemotaxis are actin polymerization (3). The membrane cytoskeleton is believed to control the lateral mobility of integral membrane proteins as well as influencing cell shape and mobility. Thus, pharmacological modulations of neutrophil chemotaxis may be related to an effect of the pharmacological agents on the membrane cytoskeleton. The present study was designed to characterize the effect of PTX …on actin polymerization of freely-suspended PMN before and after stimulation by the chemotactic factor f-MLP. We used flow cytometry to determine the proportion of actin in the filamentous form, and Rhodamine-Phalloidin as fluorescent probe (4). PTX decreased actin polymerization in response to stimulation by f-MLP. The reduction in F-actin by PTX was higher in the samples with higher activation ratios as compared with untreated PMN. Show more

Keywords: Pentoxifylline, granulocyte, actin, cytoskeleton, polymerization, rhodamine-phalloidin

DOI: 10.3233/BIR-1990-273-421

Citation: Biorheology, vol. 27, no. 3-4, pp. 445-448, 1990

Authors: Novick Jr., William J. | Sullivan, Gail | Mandell, Gerald

Article Type: Research Article

Abstract: Polymorphonuclear (PMN) overactivation plays a critical role in microcirculation as well as in conditions such as multiorgan failure (MOF). Pentoxifylline has been shown to prevent PMN activation by endotoxin and cytokines such as TNFalpha and IL-1. In addition, MOF induced by IL-2 in animals can be prevented by pentoxifylline. The present studies evaluated two aspects of PMN activation and pentoxifylline interaction. The first was the time sequence for pentoxifylline prevention of TNFalpha activation and the second was the activity of pentoxifylline on amphotericin B activation of PMNs. TNFalpha activation of PMNs is blocked by pentoxifylline when cells …are exposed to pentoxifylline prior to TNFalpha or after TNFalpha . Amphotericin B activation of PMNs was demonstrated by a decreased chemotaxis, increased chemiluminescence, and increased PMN spreading. In all conditions, pentoxifylline decreased amphotericin B activation of PMNs. These results suggest that pentoxifylline can reverse cytokine activation of PMNs and that pentoxifylline may alter some of the toxic effects of amphotericin. Show more

Keywords: Polymorphonuclear leukocytes, tissue necrosis factor (TNF), multiorgan failure (MOF)

DOI: 10.3233/BIR-1990-273-422

Citation: Biorheology, vol. 27, no. 3-4, pp. 449-454, 1990

Authors: Beretz, Alain | Klein-Soyer, Claude | Archipoff, Gisèle | Camilla, Christophe | Brisson, Christine | Freyssinet, Jean-Marie | Cazenave, Jean-Pierre

Article Type: Research Article

Abstract: Endothelial cells play an important role in the regulation of thrombosis. Normal resting (i.e. unstimulated) endothelial cells exhibit antithrombotic activity. This property is due to an active participation of endothelial cells in the inhibition of platelet adhesion and aggregation, in the inhibition of thrombin generation, in the direct inactivation of thrombin, and in clot lysis through the fibrinolytic system. When endothelial cells are stimulated by cytokines such as tumor necrosis factor (TNF) or interleukin 1 (IL-1), they may switch to an active procoagulant state. On the one hand, thrombin generation can be regulated on the endothelial cell surface by thrombomodulin, …which allows the activation by thrombin of protein C which subsequently acquires and expresses potent anticoagulant properties. On the other hand, after activation, the same endothelial cell can express tissue factor on its surface, which will lead to the triggering of the coagulation cascade resulting in the generation of thrombin. TNF has been shown both to induce tissue factor gene expression and to suppress transcription of the thrombomodulin gene in endothelial cells. Many cytokines induce tissue factor gene expression and procoagulant activity in the monocyte/macrophage lineage; they also stimulate adhesion of leukocytes to endothelial cells. Cytokines such as IL-1 or TNF can thus be characterized as important intercellular messengers during the onset of coagulation. The role of these compounds can be schematized as: 1) agents of the stimulation of endothelial cells by leukocytes, 2) agents of stimulation of leukocytes by endothelial cells, 3) localization of the coagulation response through the initiation of endothelial cell-leukocyte interactions. Pharmacological modulation of these responses is possible along two pathways: 1) inhibition of the activation of endothelial cells or leukocytes responsible for cytokine release, 2) inhibition of the cytokine-induced cellular activation responsible for potentiation of procoagulant activity. Show more

Keywords: Endothelium, leukocyte, procoagulant, anticoagulant, tissue factor, thrombomodulin

DOI: 10.3233/BIR-1990-273-423

Citation: Biorheology, vol. 27, no. 3-4, pp. 455-460, 1990

Authors: Puchelle, Edith

Article Type: Introduction

DOI: 10.3233/BIR-1990-273-424

Citation: Biorheology, vol. 27, no. 3-4, pp. 461-464, 1990

Authors: Verdugo, P. | Langley, L. | Aitkent, M.L. | Villalon, M.

Article Type: Research Article

Abstract: Mucins, are densely packed in secretory granules of goblet cells. Upon exocytosis they undergo massive hydration that results in the formation of the mucus gel. Because the mucin polymer network is held together by tangles and low energy bonds, the rheological properties of this gel are mainly determined by the degree of postexocytotic hydration. Hydration in mucus is governed by a Donnan equilibrium as it is driven by electrostatic interaction among the polyionic charges of the mucins and other fixed polyions. Although, variations of charge density of the mucin polymer could be an efficient physiologic mechanism to control the rate …of mucus hydration and rheology, this subject has not been investigated. In here we describe a primary tissue culture system of cervical goblet cells of the monkey uterus. This preparation allows to measure directly the kinetic of hydration of exocytosed mucins. Because the physicochemical parameters of the bathing medium can be effectively controlled, variations in the kinetic of mucins swelling upon exocytosis, can be used as a convenient indicator of fluctuations of charge density in secretory products. Since the cervical mucosa readily respond to endocrine influences, this preparation can provide a useful model to study the effect of hormones or other transmitters on polyionic charge density of secretory product. Show more

Keywords: mucus, mucin, hydration kinetics, cervix, goblet cell, tissue culture

DOI: 10.3233/BIR-1990-273-425

Citation: Biorheology, vol. 27, no. 3-4, pp. 465-470, 1990

Authors: Crepin, M. | Porchet, N. | Aubert, J.P. | Degand, P.

Article Type: Research Article

Abstract: A λ gt11 cDNA library constructed from human tracheal mucosa was screened with rabbit antibodies raised to chemically deglycosylated pronase glycopeptides from bronchial mucins. This library allowed us to select 20 positive clones. The total or partial nucleotide sequences of 14 of them were classified in three different types of organization or families. Antibodies purified from total immuneserum on each positive clone were specific either of goblet cells or goblet cells and mucous cells, showing a differential cellular expression of the peptide axis of mucins. The use of each type of nucleic probe in Northern blot analysis confirmed the …existence of a great heterogeneity of the RNAs contributing to a great peptide heterogeneity. Show more

Keywords: Mucins, cDNA library, peptide organization, Northern blot analysis

DOI: 10.3233/BIR-1990-273-426

Citation: Biorheology, vol. 27, no. 3-4, pp. 471-484, 1990

Authors: Basbaum, Carol | Gallup, Marianne | Gum, James | Kim, Young | Jany, Berthold

Article Type: Research Article

Abstract: The molecular mechanisms mediating mucous cell metaplasia and hypersecretion in the respiratory tract are unknown. Previous work suggests that mucous metaplasia requires the induction of mucin gene expression. We are investigating this possibility by monitoring steady state levels of mucin mRNA in a model of mucous cell metaplasia induced by SO2 exposure. Male Sprague Dawley rats were exposed to 400 ppm SO2 gas in air for 3h per day, 5 days per week for 0,1,2, or 3 weeks. Sham controls were exposed to air under similar conditions. After 3 weeks, morphological changes were apparent in the epithelium of …SO2 exposed rats at all levels from the trachea to the distal airways. The epithelial thickness increased, as well as the number and size of glands in the trachea. Epithelial mucous (goblet) cells increased from 0 to 4.5 per rom in the trachea, 0.2 to 6.2 per rom in the main stem bronchi, and 0.2 to 22.7 per rom in the distal airways (mean values obtained for 3-6 tissue blocks per airway level per condition). In parallel experiments, we used SMUC 41, a 950 bp human intestinal cDNA to isolate a human airway cDNA, HAM-1 from a cDNA library constructed in bacteriophage from human bronchial poly A+ RNA. HAM-1 is a 90 bp cDNA encoding a threonine- and proline-rich peptide with 96% homology to the human intestinal cDNA SMUC-41. Next we probed total and poly A+ airway RNA from rats in each exposure condition with SMUC-41 or HAM-1. Blots were then stripped and reprobed with a cDNA encoding beta actin. Densitometry values normalized for the amount of RNA loaded per lane (as determined by actin hybridization intensity) showed that mucin mRNA increased 8-9 fold as a function of SO2 exposure. This is consistent with the possibility that mucin gene transcription is induced by SO2 exposure, and may represent a primary event in the development of mucous metaplasia and hypersecretion. Show more

Keywords: mucous cells, mucin, differentiation, transcription, epithelium

DOI: 10.3233/BIR-1990-273-427

Citation: Biorheology, vol. 27, no. 3-4, pp. 485-489, 1990

Authors: Kim, K.C. | Singh, B.N.

Article Type: Research Article

Abstract: Confluent cultures of hamster tracheal surface epithelial (HTSE) cells are highly enriched with secretory cells and secrete mucins. Ultrastructural studies of cellular localization of these mucins show that mucins are found not only inside secretory granules but also on the apical surface of secretory cells during active secretion, and secreted mucins are highly associated with lipids. In the present communication, we analyzed lipids associated with both cellular and secreted mucins following metabolic radiolabeling of these cultured cells with [3 H]palmitic acid. We found that profiles of lipids associated with both cellular and secreted mucins are almost identical not only qualitatively …but also quantitatively. It is concluded that the lipid association with mucins seems to take place before secretion. The origin of the cell surface-bound mucins is discussed. Show more

Keywords: Mucin, lipid, tracheal, goblet cell

DOI: 10.3233/BIR-1990-273-428

Citation: Biorheology, vol. 27, no. 3-4, pp. 491-501, 1990

Authors: Baszkin, Adam | Proust, Jacques Emile | Monsenego, Phillipe | Boissonnade, Marie Martine

Article Type: Research Article

Abstract: The wettability of poly(methyl methacrylate) and polyethylene by water and aqueous mucin solutions have been studied by sessile drop and under-water captive air bubble contact angles, respectively. From the sessile drop and octane under-water contact angles the polymer-water interfaces have been characterized in terms of works of adhesion and acid-base (polar) interactions. A large water-air contact angle hysteresis observed with poly(methyl methacrylate) surfaces has been attributed to side-chain B relaxations of polymer ester methyl groups. The wettabilities of the polymers by mucin aqueous solutions have been studied as a function of protein concentration and related to the surface tensions. A …positive slope of adhesion tension vs surface tension line, characteristic of polar surfaces, was found with poly(methyl methacrylate). By contrast, a change in the slope, explained as a change in mucin relative adsorption densities at solid/liquid and solid/vapour interfaces, was observed with polyethylene. This adhesion tension behavior appeared to be in agreement with previous data we have published concerning the quantity and state of mucin which are adsorbed to polymers characterized by different surface properties. Show more

Keywords: Mucin, Wettability, Polyettlylene, Poly(methyl methacrylate), Surface Tension

DOI: 10.3233/BIR-1990-273-429

Citation: Biorheology, vol. 27, no. 3-4, pp. 503-514, 1990