Publications
87 results found
Seebach J, Mädler HJ, Wojciak-Stothard B, et al., 2005, Tyrosine phosphorylation and the small GTPase rac cross-talk in regulation of endothelial barrier function, THROMBOSIS AND HAEMOSTASIS, Vol: 94, Pages: 620-629, ISSN: 0340-6245
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- Citations: 27
Wojciak-Stothard B, Tsang LYF, Haworth SG, 2005, Rac and Rho play opposing roles in the regulation of hypoxia/reoxygenation-induced permeability changes in pulmonary artery endothelial cells, AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, Vol: 288, Pages: L749-L760, ISSN: 1040-0605
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- Citations: 142
Tsang LYF, Vallance P, Leiper J, et al., 2005, Regulatory effects of NOS inhibitors on the activity of Rho GTPases and endothelial function in pulmonary macro and microvascular endothelial cells in vitro., 3rd European Meeting on Vascular Biology and Medicine, Publisher: KARGER, Pages: 102-103, ISSN: 1018-1172
Wojciak-Stothard B, Tsang LYF, Hall S, et al., 2005, Rho GTPases Rac1 and RhoA act as regulators of endothelial phenotype and function in hypoxia-induced pulmonary hypertension of the newborn., 3rd European Meeting on Vascular Biology and Medicine, Publisher: KARGER, Pages: 89-90, ISSN: 1018-1172
Wojciak-Stothard B, Ridley AJ, 2003, Shear stress-induced endothelial cell polarization is mediated by Rho and Rac but not Cdc42 or PI 3-kinases, Journal of Cell Biology, Vol: 161, Pages: 429-439, ISSN: 0021-9525
Shear stress induces endothelial polarization and migration in the direction of flow accompanied by extensive remodeling of the actin cytoskeleton. The GTPases RhoA, Rac1, and Cdc42 are known to regulate cell shape changes through effects on the cytoskeleton and cell adhesion. We show here that all three GTPases become rapidly activated by shear stress, and that each is important for different aspects of the endothelial response. RhoA was activated within 5 min after stimulation with shear stress and led to cell rounding via Rho-kinase. Subsequently, the cells respread and elongated within the direction of shear stress as RhoA activity returned to baseline and Rac1 and Cdc42 reached peak activation. Cell elongation required Rac1 and Cdc42 but not phosphatidylinositide 3-kinases. Cdc42 and PI3Ks were not required to establish shear stress–induced polarity although they contributed to optimal migration speed. Instead, Rho and Rac1 regulated directionality of cell movement. Inhibition of Rho or Rho-kinase did not affect the cell speed but significantly increased cell displacement. Our results show that endothelial cells reorient in response to shear stress by a two-step process involving Rho-induced depolarization, followed by Rho/Rac-mediated polarization and migration in the direction of flow.
Wojciak-Stothard B, Ridley AJ, 2002, Rho GTPases and the regulation of endothelial permeability, VASCULAR PHARMACOLOGY, Vol: 39, Pages: 187-199, ISSN: 1537-1891
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- Citations: 381
Hall SM, Wojciak-Stothard B, Haworth SG, 2001, Pulmonary artery smooth muscle cells of different developmental lineages have distinct structural and functional phenotypes, MOLECULAR BIOLOGY OF THE CELL, Vol: 12, Pages: 510A-511A, ISSN: 1059-1524
Wójciak-Stothard B, Potempa S, Eichholtz T, et al., 2001, Rho and Rac but not Cdc42 regulate endothelial cell permeability, JOURNAL OF CELL SCIENCE, Vol: 114, Pages: 1343-1355, ISSN: 0021-9533
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- Citations: 391
Wojciak-Stothard B, Williams L, Ridley AJ, 1999, Monocyte adhesion and spreading on human endothelial cells is dependent on Rho-regulated receptor clustering, The Journal of Cell Biology, Vol: 145, Pages: 1293-1307, ISSN: 0021-9525
The GTPase Rho is known to mediate the assembly of integrin-containing focal adhesions and actin stress fibers. Here, we investigate the role of Rho in regulating the distribution of the monocyte-binding receptors E-selectin, ICAM-1, and VCAM-1 in human endothelial cells. Inhibition of Rho activity with C3 transferase or N19RhoA, a dominant negative RhoA mutant, reduced the adhesion of monocytes to activated endothelial cells and inhibited their spreading. Similar effects were observed after pretreatment of endothelial cells with cytochalasin D. In contrast, dominant negative Rac and Cdc42 proteins did not affect monocyte adhesion or spreading. C3 transferase and cytochalasin D did not alter the expression levels of monocyte-binding receptors on endothelial cells, but did inhibit clustering of E-selectin, ICAM-1, and VCAM-1 on the cell surface induced by monocyte adhesion or cross-linking antibodies. Similarly, N19RhoA inhibited receptor clustering. Monocyte adhesion and receptor cross-linking induced stress fiber assembly, and inhibitors of myosin light chain kinase prevented this response but did not affect receptor clustering. Finally, receptor clusters colocalized with ezrin/moesin/ radixin proteins. These results suggest that Rho is required in endothelial cells for the assembly of stable adhesions with monocytes via the clustering of monocyte-binding receptors and their association with the actin cytoskeleton, independent of stress fiber formation.
Wójciak-Stothard B, Entwistle A, Garg R, et al., 1998, Regulation of TNF-α-induced reorganization of the actin cytoskeleton and cell-cell junctions by Rho, Rac, and Cdc42 in human endothelial cells, JOURNAL OF CELLULAR PHYSIOLOGY, Vol: 176, Pages: 150-165, ISSN: 0021-9541
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- Citations: 340
Wòjciak-Stothard B, Denyer M, Mishra M, et al., 1997, Adhesion, orientation, and movement of cells cultured on ultrathin fibronectin fibers., In Vitro Cell Dev Biol Anim, Vol: 33, Pages: 110-117, ISSN: 1071-2690
This study examined the behavior of rat tendon fibroblasts, baby hamster kidney fibroblasts, macrophage-like P388D1 cells, and neurons from rat dorsal root ganglia, cultured on fibronectin strands 0.2-5 micrograms in diameter. We investigated cell spreading, orientation, formation of focal contacts, the speed of cell movement, and the speed of neurite outgrowth in cells cultured on fibronectin strands, glass covered with fibronectin, and plain, nontreated glass. Fibronectin strands significantly promoted cell spreading and caused a marked alignment of all kinds of cells to the direction of the fiber. The fibers caused the alignment of actin filaments in fibroblasts and focal contacts in fibroblasts and macrophages and increased polymerization of F-actin in cells. Fibronectin fibers also increased the speed and persistence of cell movement and the rate of neurite outgrowth. Macrophages grown on fibronectin fibers produced numerous actin-rich microspikes and adopted a polarized, migratory phenotype. These findings indicate that fibronectin strands, resembling natural components of the extracellular matrix, are more effective in activating various types of cells then two-dimensional, fibronectin-covered substrata. The results also confirm the suitability of the three-dimensionally oriented fibronectin form for use in clinical practice.
Wójciak-Stothard B, Curtis A, Monaghan W, et al., 1996, Guidance and activation of murine macrophages by nanometric scale topography., Exp Cell Res, Vol: 223, Pages: 426-435, ISSN: 0014-4827
We studied the guidance and activation of macrophages from the P388D1 cell line and rat peritoneum by topographic features on a nanometric scale. Cells were plated on plain fused silica substrata or substrata with microfabricated grooves and steps, 30-282 nm deep. The contact of cells with the patterned surface activated cell spreading and adhesion and increased the number of protrusions of the cell membrane. These changes were accompanied by an increase in the amount of F-actin in cells. The accumulation of F-actin and vinculin in cells was observed along the edges of single steps or grooves. Formation of focal contacts along discontinuities in the substratum was accompanied by the phosphorylation of tyrosine colocalized with F-actin and vinculin. A similar pattern of staining was seen in cells stained for vitronectin receptor, alphaV integrin, but not for integrins alpha5beta1 or alpha3beta1. Cells cultured on nanogrooves showed a higher phagocytotic activity than cells cultured on plain substrata. We show that macrophages can react to ultrafine features of topography of a size comparable to that of a single collagen fiber and become activated by the contact with topographic features.
Wójciak-Stothard B, Madeja Z, Korohoda W, et al., 1995, Activation of macrophage-like cells by multiple grooved substrata. Topographical control of cell behaviour., Cell Biol Int, Vol: 19, Pages: 485-490, ISSN: 1065-6995
We studied the influence of substrata topography on the behaviour of murine P388D1 macrophage cell line. Cells were plated on plain fused silica substrata or substrata with microfabricated grooves of varying depth and width. Cell spread area, elongation, orientation and F-actin content were measured on plain substratum and 6 sets of gratings. The speed and persistence of cell movement were also studied. We found that patterned substrata substantially activated cell spreading and elongation and significantly increased the persistence and speed of cell movement, shallow grooves being more effective than deep ones. The contact of cells with micropatterned substrata significantly increased the F-actin content in cells. The sensitivity of LPS (lipopolisaccharide) stimulated and unstimulated macrophages to topographical cues was also compared.
WOJCIAK B, CROSSAN J, CURTIS ASG, et al., 1995, GROOVED SUBSTRATA FACILITATE IN-VITRO HEALING OF COMPLETELY DIVIDED FLEXOR TENDONS, JOURNAL OF MATERIALS SCIENCE-MATERIALS IN MEDICINE, Vol: 6, Pages: 266-271, ISSN: 0957-4530
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- Citations: 40
Wójciak-Stothard B, Curtis AS, Monaghan W, et al., 1995, Role of the cytoskeleton in the reaction of fibroblasts to multiple grooved substrata., Cell Motil Cytoskeleton, Vol: 31, Pages: 147-158, ISSN: 0886-1544
The role of the cytoskeleton and cell attachments in the alignment of baby hamster kidney fibroblasts to ridge and groove substratum topography was investigated using confocal scanning microscopy. This was carried out with normal cells and cells treated with the cytoskeleton modifiers cytochalasin D, colcemid, and taxol. Actin was localised with fluorescent phalloidin. Tubulin, vinculin, and intracellular adhesion molecule-1 were visualised by indirect immunofluorescence. The spreading, elongation, and orientation of the cells after 24 h of culture in these conditions were measured on grooves of 5, 10, and 25 microns width and 0.5, 1, 2, and 5 microns depth. We have also observed events over the first 30 min of cell attachment. Five minutes after cell attachment, F-actin condensations were seen close to the intersection of groove wall and ridge top, that is, at a topographic discontinuity. The condensations were often at right angles to the groove edge and showed a periodicity of 0.6 microns. Vinculin arrangement at the early stages of cell spreading was similar to that of actin. Organisation of the microtubule system followed later, becoming obvious at about 30 min after cell plating. The Curtis and Clark theory (that cells react to topography primarily at lines of discontinuity in the substratum by actin nucleation) is supported by these results. The use of cytoskeletal poisons did not entirely abolish cell reaction to grooves. Colcemid increased cell spreading and reduced cell orientation and elongation. Cytochalasin D reduced cell spreading, orientation, and elongation. Taxol reduced cell elongation but did not affect cell spreading and orientation. We conclude that the aggregation of actin along groove/ridge boundaries is a primary driving event in determining fibroblast orientation on microgrooved substrata.
WOJCIAK B, CROSSAN JF, 1994, THE EFFECTS OF T-CELLS AND THEIR PRODUCTS ON IN-VITRO HEALING OF EPITENON CELL MICROWOUNDS, IMMUNOLOGY, Vol: 83, Pages: 93-98, ISSN: 0019-2805
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- Citations: 30
CURTIS ASC, WILKINSON CWD, WOJCIAK B, 1994, COMPARING THE EFFECTS OF TOPOGRAPHY AND CHEMISTRY ON CELL-BINDING TO SUBSTRATES, ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, Vol: 207, Pages: 269-COLL, ISSN: 0065-7727
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- Citations: 1
KOROHODA W, WOJCIAK B, BEREITERHAHN J, 1994, AN IMPROVED METHOD FOR THE ISOLATION OF CELLS FROM TISSUES - PREVENTION OF CELL ROUNDING ENHANCES SPREADING AND LOCOMOTION OF KERATINOCYTES (EPITHELIOCYTES), FOLIA HISTOCHEMICA ET CYTOBIOLOGICA, Vol: 32, Pages: 25-30, ISSN: 0015-5586
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- Citations: 2
Korohoda W, Wójciak B, Bereiter-Hahn J, 1994, An improved method for the isolation of cells from tissues: prevention of cell rounding enhances spreading and locomotion of keratinocytes (epitheliocytes)., Folia Histochem Cytobiol, Vol: 32, Pages: 25-30, ISSN: 0239-8508
Benzamide (10mM) in Ca-free solution anesthetizes small cold blooded animals such as tadpoles of Rana temporaria, Xenopus laevis and an aquarium fish, guppy (Lebistes [Poecilia] reticulatus). Trypsinization of tails of tadpoles in Ca-free, benzamide-supplemented solution permits isolation of keratinocytes (epitheliocytes) maintaining polygonal shapes and prevents cell rounding. Such cells quickly attach to glass, spread and commence locomotion in contrast to those cells which assume spherical shape after standard trypsinization.
WOJCIAK B, CROSSAN JF, 1993, THE ACCUMULATION OF INFLAMMATORY CELLS IN SYNOVIAL SHEATH AND EPITENON DURING ADHESION FORMATION IN HEALING RAT FLEXOR TENDONS, CLINICAL AND EXPERIMENTAL IMMUNOLOGY, Vol: 93, Pages: 108-114, ISSN: 0009-9104
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- Citations: 39
Wojciak B, Crossan JF, 1993, The accumulation of inflammatory cells in synovial sheath and epitenon during adhesion formation in healing rat flexor tendons., Clin Exp Immunol, Vol: 93, Pages: 108-114, ISSN: 0009-9104
The accumulation of inflammatory cells in synovial tissue was studied using indirect immunofluorescence assays on cell cultures and frozen tissue sections of healing rat digital flexor tendons. Flexor tendons were collected from rats 3, 7 and 14 days after crush injury. Tendon sheath and epithenon cells were isolated by sequential enzymic digestion and cultured for 2 days. Subpopulations of synovial and inflammatory cells were identified with MoAbs against cell surface glycoproteins present on B lymphocytes (CD45), T lymphocytes (CD2, CD4, CD8), macrophages (CD14) and endothelial cells. A phagocytosis assay was also used to identify macrophages. We report a substantial increase in the number of T lymphocytes (mainly helper/inducer) and phagocytotic cells with monocyte/macrophage surface markers in tendon sheath and epitenon 3 days after crush injury. The infiltration of inflammatory cells into synovial sheath and epitenon preceded an increase in fibronectin production by tendon cells which was seen 7 days after injury. To study the interaction between T lymphocytes and synovial cells in vitro, we established synovial fibroblast-like type B cell cultures and used stimulated and non-stimulated T lymphocytes in cell binding assays. We observed increased adhesiveness between unstimulated synovial cells and synovial cells previously cultured with activated and non-activated T lymphocytes. ELISA inhibition studies have shown an increase in fibronectin production by synovial fibroblasts co-cultured with stimulated CD4+ T lymphocytes. We suggest that the presence of inflammatory cells in synovial sheath and epitenon during tendon healing induces synovial fibroblasts and epitenon cells to increase their production of fibronectin, which provides a scaffold for subsequent adhesion formation.
BAIRD KS, ALWAN WH, CROSSAN JF, et al., 1993, T-CELL-MEDIATED RESPONSE IN DUPUYTRENS DISEASE, LANCET, Vol: 341, Pages: 1622-1623, ISSN: 0140-6736
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- Citations: 33
Baird KS, Alwan WH, Crossan JF, et al., 1993, T-cell-mediated response in Dupuytren's disease., Lancet, Vol: 341, Pages: 1622-1623, ISSN: 0140-6736
The cause of Dupuytren's disease is unknown, but inflammatory cells might have a role. Enzymatic digestion of diseased tissue permits identification and immunofluorescent labelling of a cell subset displaying inflammatory cell morphology. Cytofluorimetry of this cell population demonstrated the presence of CD3-positive lymphocytes and expression of major histocompatibility complex (MHC) class II proteins. These results raise the possibility that Dupuytren's disease is a T-cell-mediated autoimmune disorder. The development of medical treatment on this basis may reduce the need for surgery, with its associated morbidity and high recurrence rates.
Wójciak B, 1993, Ehrlich ascites tumour cell spreading is induced upon contact with immobilized isolated fibroblast plasma membranes and fibronectin., Folia Histochem Cytobiol, Vol: 31, Pages: 27-33, ISSN: 0239-8508
Ehrlich ascites tumour (EAT) cells normally growing in suspension, adhere and spread upon contact with fibroblasts. Changes in EAT cell morphology and adhesiveness induced in contact with purified fibroblast plasma membranes, fibroblast extracellular matrix and fibronectin were investigated. EAT cells adhered readily to glass covered with fibroblast extracellular matrix or fibronectin, but they did not spread. Immobilization of fibroblast plasma membranes or fibronectin on nitrocellulose restored EAT cell ability to spread. Tumour cells spread on immobilized plasma membranes or fibronectin maintained a "fried egg" morphology, while these spread on living fibroblasts were rather elongated. EAT cells preincubated with anti-fibronectin serum lost their ability to adhere to any of the tested substrata: living and fixed fibroblasts, fibroblast extracellular matrix, nitrocellulose with immobilized fibroblast plasma membranes or fibronectin. When anti-fibronectin serum was applied only to these substrata and it was washed out before EAT cell plating, EAT cell adhesion and spreading were not significantly changed. Extraction of extracellular matrix components from immobilized fibroblast plasma membranes did not influence EAT adhesion and spreading. The results are discussed with reference to the specific role of fibronectin in the growth of EAT cells as a solid tumour or suspension.
WOJCIAK B, 1993, EHRLICH ASCITES TUMOR-CELL SPREADING IS INDUCED UPON CONTACT WITH IMMOBILIZED ISOLATED FIBROBLAST PLASMA-MEMBRANES AND FIBRONECTIN, FOLIA HISTOCHEMICA ET CYTOBIOLOGICA, Vol: 31, Pages: 27-33, ISSN: 0015-5586
KOROHODA W, WOJCIAK B, 1992, CELL-SHAPE AND CELL-ADHESION, 11TH SCHOOL ON BIOPHYSICS OF MEMBRANE TRANSPORT, Publisher: AGRICULTURAL UNIV WROCLAW DEPT PHYSICS & BIOPHYSICS, Pages: A291-A304
WOJCIAK B, KOROHODA W, 1990, EHRLICH ASCITES TUMOR-CELLS SHOW TISSUE-SPECIFIC ADHERENCE AND MODIFY THEIR SHAPE UPON CONTACT WITH EMBRYONIC FIBROBLASTS AND MYOTUBES, JOURNAL OF CELL SCIENCE, Vol: 97, Pages: 433-438, ISSN: 0021-9533
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- Citations: 11
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