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PubMed1999

RGD-CAP ((beta)ig-h3) enhances the spreading of chondrocytes and fibroblasts via integrin alpha(1)beta(1).

In previous studies, RGD-CAP (collagen-associated protein containing the RGD sequence) isolated from a collagen fiber-rich fraction of pig cartilage was found to be orthologous to human (beta)ig-h3, which is synthesized by lung adenocarcinoma cells in response to transforming growth factor-beta. In the present study, we examined the effect of recombinant chick RGD-CAP on the spreading of chondrocytes and fibroblasts using RGD-CAP-coated dishes. When rabbit articular chondrocytes, chick embryonic sternal chondrocytes, rabbit peritoneal fibroblasts or human MRC5 fibroblasts were seeded on plastic dishes coated with RGD-CAP, cell spreading was enhanced compared with that on control dishes (bovine serum albumin- or beta-galactosidase-coated dishes). The effect of RGD-CAP on the cell spreading required divalent cations (Mg(2+) or Mn(2+)), and was reduced by EDTA. Monoclonal antibodies (mAbs) to the human integrin alpha(1) or beta(1) subunit, but not to the alpha(2), alpha(3), alpha(5) or beta(2) subunits, suppressed the RGD-CAP-induced spreading of human MRC5 fibroblasts. In a parallel experiment, the mAb to the alpha(5) subunit, but not the mAb to the alpha(1) subunit, suppressed fibronectin-induced spreading of these cells. These findings suggest that RGD-CAP is a novel ligand for integrin alpha(1)beta(1) that dose not bind to the RGD motif. Accordingly, an RGD-CAP fragment, which carries a deletion in the C-terminal region containing the RGD motif, was still capable of stimulating cell spreading.

Animals↗
PubMed1999

Beta1-integrins mediate Ca2+-signalling and T cell spreading via divergent pathways.

Interaction of Jurkat T-lymphocytes with two extracellular matrix (ECM) proteins of the basement membrane, laminin or collagen type IV, combined with poly-L-lysine resulted in a strong adhesion, a highly increased intracellular Ca2+-concentration ([Ca2]i), as compared to cells on laminin or collagen type IV alone and in spreading of the cells. The strong adhesion was independent of an increase in [Ca2+]i, was not mediated by a beta1-integrin, and was due to charge interaction between the positively charged polyaminoacid and the negatively charged cell surface. The latter was confirmed by substitution of poly-L-lysine by other positively charged polyaminoacids. In contrast, Ca+-signalling and spreading of the cells adhering to laminin or collagen type IV combined with poly-L-lysine was completely blocked by anti-beta1 mAb. However, spreading of the cells was independent of an increase in [Ca2+]i suggesting divergent signal transduction pathways leading to Ca2+-signalling and spreading of the cells. We elucidated these signal transduction pathways by inhibition of key enzymes involved. The tyrosine kinase inhibitor genistein blocked Ca2+-signalling as well as spreading, whereas inhibitors of PKC (calphostin C, GF109203x), PLCgamma (U73122) and PLA2 (bromophenacyl-bromide (BPB), 3-[4-octadecyl)benzoyl]acrylic acid (OBAA)) selectively blocked spreading of the cells.

Calcium Signaling↗
PubMed1998

Type I collagen synthesis in cultured human fibroblasts: regulation by cell spreading, platelet-derived growth factor and interactions with collagen fibers.

Type I collagen protein and pro-alpha 1(I) collagen mRNA levels were investigated in human dermal fibroblasts cultured on substrates which induced distinct morphologies. Induction of type I collagen protein synthesis required cell spreading in monolayer cultures; mere attachment to dishes coated with 2-hydroxyethyl methacrylate (poly(HEMA)) did not suffice. Spread cells or round cells cultured on poly(HEMA) differed in collagen type I production, but pro-alpha 1(I) collagen mRNA levels were similar. Recombinant human platelet-derived growth factor (PDGF)-BB could replace cell spreading as a stimulus for collagen synthesis in cells cultured on poly(HEMA). At later time points, pro-alpha 1(I) collagen mRNA levels were down-regulated, although relatively less than type I collagen synthesis. Type I collagen synthesis by fibroblasts cultured in three-dimensional collagen gels was strongly down-regulated at both the protein and RNA levels. In addition to its capacity to stimulate collagen synthesis, PDGF-BB induced elongation and the formation of long processes by fibroblasts cultured in collagen gels. The stimulatory effect by cell spreading and PDGF-BB on collagen synthesis was inhibited by the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002. However, inhibition of PI3K only inhibited induction of collagen synthesis by actively spreading cells or by PDGF-BB and did not induce a down-regulation of collagen synthesis in cells which had already spread. These data demonstrate that type I collagen protein synthesis is partly independent of pro-alpha 1(I) collagen mRNA levels but highly regulated by cell shape, although this could be decoupled by PDGF-BB. Both cell shape- and PDGF-BB-induced stimulation of collagen type I synthesis depends on a signalling pathway involving PI3K. Furthermore, levels of pro-alpha 1(I) collagen mRNA in fibroblasts are partly cell shape independent but are down-regulated by fibroblast interactions with native collagen fibers.

Adult↗
PubMed1996

Reduction of Thermal Spread and Tissue Adherence with an Irrigating Bipolar Grasper

Bipolar energy heats tissues and leads to thermal spread beyond the target tissues. As temperatures rise above 70° C, tissues stick to the metal jaws of the graspers. The use of irrigating fluid while applying the energy should reduce these effects by cooling the surrounding tissues, and also permit the surgeon to clear the operative field of blood. An in vivo animal study was performed to measure these effects. Energy was applied to porcine uterine horn with and without irrigation with a new irrigating bipolar grasper. Lateral thermal spread and tissue adherence were measured. Tissue was excised and thermal spread measured grossly and microscopically. Results are shown in the table. Without irrigation, thermal spread stops at 10 seconds due to complete desiccation of the tissues. Lateral thermal spread is reduced below 10 seconds with irrigation, but it continues to rise as energy is applied. Coagulated tissues adhered to the grasper jaws in all of the nonirrigated tests. Cooling the jaws with irrigation eliminated this. The ability to irrigate through the bipolar grasper improves the surgeon's efficiency by reducing tissue sticking, and adds control by reducing thermal spread during normal application of energy.

Journal Article↗
PubMed1996

Effect of injection speed on the spread of ethanol during experimental liver ethanol injections.

RATIONALE AND OBJECTIVES: Percutaneous ethanol injection therapy may cause serious complications, most likely due to the uncontrolled spread of ethanol. The authors changed the speed of the injection into postmortem pig livers and examined the adverse spread of ethanol from the injection site and the shape and size of the resulting lesion. METHODS: One milliliter of 96% ethanol was injected into pig livers under sonographic guidance at different speeds (0.075-0.91 mL/s). The spread (graded from I to III) and the volumes and shapes of the resulting lesions seen after dissection were recorded and correlated with the injection speed. RESULTS: When increasing the speed of the injections the large, grade III spread increased significantly (P < .01). The lesions created by more rapid injection were less spherical than were those created by slower injections (P = .08). The volumes of the lesions were not affected by the injection speed. CONCLUSION: This experimental model suggests that in percutaneous ethanol injection therapy, rapid injection (> 0.3-0.4 mL/s) should be avoided to reduce the uncontrolled spread of ethanol. Spherical spreading of ethanol around the needle tip is best achieved with slow injection.

Animals↗
PubMedDate not supplied

Spatiotemporal structure of depolarization spread in cortical pyramidal cell populations evoked by diffuse retinal light flashes.

The spatiotemporal structure of cortical activity evoked by diffuse light flashes was investigated in an isolated eyecup-brain preparation of the pond turtle, Pseudemys scripta. By combining a photomicroscopic image of the preparation with voltage-sensitive dye signals recorded by a 464-element photodiode array, the spread of depolarization within different cortical areas could be directly visualized with millisecond temporal resolution. Diffuse stimulation of the contralateral eyecup initially depolarized the visual cortex at the junction between its lateral and medial divisions in a small area rostral of the ventricular eminence. From this point, the depolarization spread at different velocities (10-100 microm/ms) depending upon the direction of travel. Since the initial depolarization was always in the rostral pole, the largest spread invariably occurred in a rostral --> caudal direction. Within the confines of the medial visual cortex, depolarization spread at a constant velocity but slowed after entering the adjoining medial cortex. Increasing the stimulus illuminance increased the velocity of spread. Rostrocaudal spread of depolarization was also observed in response to electrical stimulation of the geniculocortical pathway and by direct focal stimulation of the cortical sheet. These data suggest that excitatory connections between pyramidal cell clusters play a prominent role in the initial activation of the cortex by diffuse retinal stimulation.

Animals↗
PubMed2005

Novel two-dimensional "ring and chain" morphologies in Langmuir-Blodgett monolayers of PS-b-PEO block copolymers: effect of spreading solution concentration on self-assembly at the air-water interface.

A polystyrene-b-poly(ethylene oxide) (PS-b-PEO) (MW = 141k, 11.4 wt% PEO) diblock copolymer in the hydrophobic regime was spread from chloroform solutions of various concentrations at the air-water interface, and the resultant monolayers were transferred to glass substrates and imaged using atomic force microscopy. Monolayers prepared under identical conditions were also characterized at the air-water interface via Langmuir compression isotherms. The effects of spreading solution concentration on surface features, compressibility, and limiting mean molecular area were determined, revealing several interesting trends that have not been reported for other systems of PS-b-PEO. Spreading solutions > or = 0.50 mg/mL resulted almost exclusively in dot and spaghetti morphologies, with no observed continent features, which have been commonly found in more hydrophobic systems. For lower spreading solutions, < or = 0.25 mg/mL, we observed a large predominance of two novel surface morphologies, nanoscale rings and chains. The surface pressure (pi)-area (A) isotherms also exhibited a unique dependence on the spreading solution concentration, with limiting mean molecular areas and isothermal compressibilities of PS-b-PEO monolayers increasing below a critical concentration of spreading solution, suggesting a greater contribution from the PEO blocks. These results suggest that PS chain entanglement prior to solvent evaporation plays an important kinetic role in the extent of PEO adsorption at the air-water interface and in the morphologies of the resulting self-assembled surface aggregates.

Journal Article↗
PubMed2005

Spreading kinetics of water drops on self-assembled monolayers of thiols: significance of inertial effects.

Spreading of 5-15 microL water drops on self-assembled monolayers of 1-hexadecanethiol and 11-mercapto-1-undecanol, both homogeneous and mixed compositions, formed on gold-coated silicon wafers or glass slides was recorded with a high-speed video camera. The time (t) evolution of the drop base diameter (D) during spreading was analyzed by a power law-correlation: D approximately t(n). The n value was found to increase from n = 0.3-0.5 for water drops on hydrophobic surfaces characterized by the advancing water contact angle of thetaA = 94-104 degrees to n = 0.5-0.8 on less hydrophobic surfaces (thetaA = 45-66 degrees ). These experimental values were found to be of similar magnitude as the literature values reported for small drops and bubbles, which spread over a variety of different substrates including water and water-ethanol drops on self-assembled monolayers of alkylsilanes, air bubbles in water on glass, molten metals on solid metals and ceramics, hydrocarbon drops on water, and others. Inertial effects, which are often not accounted for in the analysis of spreading results, appear to have an impact on the spreading kinetics of small drops in at least the first few milliseconds of the spreading phenomenon.

Journal Article↗
PubMed2006

Liquid nanodroplets spreading on chemically patterned surfaces.

Controlling the spatial distribution of liquid droplets on surfaces via surface energy patterning can be used to deliver material to specified regions via selective liquid/solid wetting. Although studies of the equilibrium shape of liquid droplets on heterogeneous substrates exist, much less is known about the corresponding wetting kinetics. Here we present large-scale atomistic simulations of liquid nanodroplets spreading on chemically patterned surfaces. Results are presented for lines of polymer liquid (droplets) on substrates consisting of alternating strips of wetting (equilibrium contact angle theta0 = 0 degrees) and nonwetting (theta0 approximately 90 degrees) material. Droplet spreading is compared for different wavelength lambda of the pattern and strength of surface interaction on the wetting strips. For small lambda, droplets partially spread on both the wetting and nonwetting regions of the substrate to attain a finite contact angle less than 90 degrees. In this case, the extent of spreading depends on the interaction strength in the wetting regions. A transition is observed such that, for large lambda, the droplet spreads only on the wetting region of the substrate by pulling material from nonwetting regions. In most cases, a precursor film spreads on the wetting portion of the substrate at a rate strongly dependent on the width of the wetting region.

Journal Article↗
PubMed1995

Radial spread of sodium lauryl sulfate after topical application.

PURPOSE: Since topical application of sodium lauryl sulfate (SLS) has been reported to elevate transepidermal water loss and decrease skin capacitance in areas immediately adjacent to the applied site, studies were carried out to quantify the extent of radial spread of SLS below a topically exposed site in a hairless rat model. METHODS: Fixed sites were demarcated and the levels of SLS measured around the applied site in the epidermis, dermis and the subcutaneous tissues. Underlying deep tissue penetration and radial spread of SLS in the presence and absence of a vasoconstrictor, phenylephrine, was also quantified. RESULTS: In a typical 24 hour study, the radial spread of SLS was observed to a distance of approximately 0.75 cm from the applied site. The use of phenylephrine (1:20000), did not significantly enhance either the local underlying tissue (apart from underlying epidermis) concentration or radial spread of SLS relative to no vasoconstrictor treatment. CONCLUSIONS: Given that SLS impairs barrier function of the skin, its radial spread could be explained by a passive diffusion process. Vasoconstrictor did not remarkably alter SLS penetration and radial spread possibly due to the competing effects of vasodilation (caused by SLS) and vasoconstriction (caused by phenylephrine).

Administration, Topical↗
PubMed2002

Guilty by mere association: evaluative conditioning and the spreading attitude effect.

Five experiments investigated the phenomenon that attitude formation is not confined to the co-occurrence of an attitudinal object with an evaluated experience. The pairing of a target with a (dis)liked person not only affects the evaluation of the previously neutral person but spreads to other individuals who are (pre)associated with the target (spreading attitude effect). Experiments 1 and 2 provided evidence for the spreading attitude effect in appetitive as well as aversive evaluative conditioning. Experiment 3 showed that the spreading attitude effect is a robust phenomenon resistant to extinction. Experiment 4 demonstrated that attitude spread can be transferred to 2nd-order conditioning. Finally, Experiment 5 supports the notion that the spreading attitude effect is not dependent on cognitive resources. Implications for social as well as applied psychology are discussed.

Adult↗
PubMed2001

Evidence of recent volcanic activity on the ultraslow-spreading Gakkel ridge.

Seafloor spreading is accommodated by volcanic and tectonic processes along the global mid-ocean ridge system. As spreading rate decreases the influence of volcanism also decreases, and it is unknown whether significant volcanism occurs at all at ultraslow spreading rates (<1.5 cm yr(-1)). Here we present three-dimensional sonar maps of the Gakkel ridge, Earth's slowest-spreading mid-ocean ridge, located in the Arctic basin under the Arctic Ocean ice canopy. We acquired this data using hull-mounted sonars attached to a nuclear-powered submarine, the USS Hawkbill. Sidescan data for the ultraslow-spreading (approximately 1.0 cm yr(-1)) eastern Gakkel ridge depict two young volcanoes covering approximately 720 km2 of an otherwise heavily sedimented axial valley. The western volcano coincides with the average location of epicentres for more than 250 teleseismic events detected in 1999, suggesting that an axial eruption was imaged shortly after its occurrence. These findings demonstrate that eruptions along the ultraslow-spreading Gakkel ridge are focused at discrete locations and appear to be more voluminous and occur more frequently than was previously thought.

Journal Article↗
PubMed1984

Extracellular pH changes during spreading depression and cerebral ischemia: mechanisms of brain pH regulation.

We have examined the extracellular pH (pHe) during spreading depression and complete cerebral ischemia in rat parietal cortex utilizing double-barrelled H+ liquid ion exchanger microelectrodes. The baseline pHe of the parietal cortex was 7.33 at a mean arterial PCO2 of 38 mm Hg. Following spreading depression and cerebral ischemia, highly reproducible triphasic changes in pHe occurred, which were intimately related to the negative deflection in tissue potential (Ve). The changes in pHe for spreading depression (n = 23) were a small initial acidic shift, beginning before the rapid change in Ve, followed by a rapid transient alkaline shift of 0.16 pH units, the onset of which coincided with the negative deflection in Ve. A prolonged acidic shift of 0.42 pH units then occurred. The maximal decrease in pHe was to 6.97 and the mean duration of the triphasic pHe change was 7.8 min. The lactate concentration in brain cortex increased from baseline 1.2 mM to 7.0 mM (n = 6) during the maximal acidic change in spreading depression. In addition, lactate levels correlated well with resolution of the pHe changes during spreading depression. The triphasic pHe changes following complete cerebral ischemia were an initial acidic shift of 0.43 pH units which developed over 2 min, then an alkaline shift of 0.10 pH units coincident with the negative deflection in Ve, and a final acidic shift of 0.26 pH units. The terminal pHe was 6.75. Superfusion of the cortex with inhibitors of carbonic anhydrase (acetazolamide), Na+/H+ counter transport (amiloride), and Cl-/HCO-3 countertransport (4,4'-diisothiocyanostilbene-2,2'-disulfonic acid) altered the triphasic pHe changes in a similar fashion for both spreading depression and cerebral ischemia, providing insights into the pHe regulatory mechanisms in mammalian brain.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed2003

Spreading of nanofluids on solids.

Suspensions of nanometre-sized particles (nanofluids) are used in a variety of technological contexts. For example, their spreading and adhesion behaviour on solid surfaces can yield materials with desirable structural and optical properties. Similarly, the spreading behaviour of nanofluids containing surfactant micelles has implications for soil remediation, oily soil removal, lubrication and enhanced oil recovery. But the well-established concepts of spreading and adhesion of simple liquids do not apply to nanofluids. Theoretical investigations have suggested that a solid-like ordering of suspended spheres will occur in the confined three-phase contact region at the edge of the spreading fluid, becoming more disordered and fluid-like towards the bulk phase. Calculations have also suggested that the pressure arising from such colloidal ordering in the confined region will enhance the spreading behaviour of nanofluids. Here we use video microscopy to demonstrate both the two-dimensional crystal-like ordering of charged nanometre-sized polystyrene spheres in water, and the enhanced spreading dynamics of a micellar fluid, at the three-phase contact region. Our findings suggest a new mechanism for oily soil removal--detergency.

Journal Article↗
PubMed2004

Spreading-rate dependence of melt extraction at mid-ocean ridges from mantle seismic refraction data.

A variety of observations indicate that mid-ocean ridges produce less crust at spreading rates below 20 mm yr(-1) (refs 1-3), reflecting changes in fundamental ridge processes with decreasing spreading rate. The nature of these changes, however, remains uncertain, with end-member explanations being decreasing shallow melting or incomplete melt extraction, each due to the influence of a thicker thermal lid. Here we present results of a seismic refraction experiment designed to study mid-ocean ridge processes by imaging residual mantle structure. Our results reveal an abrupt lateral change in bulk mantle seismic properties associated with a change from slow to ultraslow palaeo-spreading rate. Changes in mantle velocity gradient, basement topography and crustal thickness all correlate with this spreading-rate change. These observations can be explained by variations in melt extraction at the ridge, with a gabbroic phase preferentially retained in the mantle at slower spreading rates. The estimated volume of retained melt balances the approximately 1.5-km difference in crustal thickness, suggesting that changes in spreading rate affect melt-extraction processes rather than total melting.

Journal Article↗
PubMed2001

C-terminal heparin-binding domain of fibronectin regulates integrin-mediated cell spreading but not the activation of mitogen-activated protein kinase.

Fibronectin (FN) stimulates multiple signalling events including mitogen-activated protein kinase (MAPK) activation. During cell spreading, both the cell-binding domain and the C-terminal heparin-binding domain (HepII) of FN co-operatively regulate cytoskeleton organization. However, in comparison with the large number of studies on the functions of cell-binding domain, there is little information about the role of HepII. We therefore investigated the effect of HepII on integrin-mediated cell spreading and adhesion on FN and MAPK activation. In contrast with cells on FN substrates, rat embryo fibroblasts on FN120, which lacks HepII, were less spread, had weaker adhesion to FN and failed to form focal adhesions and actin stress fibres. Phosphotyrosine was present in the focal contacts of rat embryo fibroblasts on FN within 30 min but was absent from cells on FN120. Overall, tyrosine phosphorylation was much less in cell lysates from cells on FN120, with decreased phosphorylation of focal adhesion kinase ('pp125FAK') on tyrosine-397, implying additional regulation of tyrosine phosphorylation by HepII. Nevertheless, adhesion-mediated MAPK activity was similar in cells on FN and on FN120. Furthermore, cells spread on FN and on FN120 substrates showed similar MAPK activation in response to treatment with epidermal growth factor and with platelet-derived growth factor. Consistently, overexpression of syndecan-4, which binds to HepII, enhanced cell spreading and adhesion on FN but did not affect integrin-mediated MAPK activation. We therefore conclude that both HepII and syndecan-4 regulate integrin-mediated cell spreading but not MAPK activation.

Actins↗
PubMed2005

Sprouty-4 negatively regulates cell spreading by inhibiting the kinase activity of testicular protein kinase.

TESK1 (testicular protein kinase 1) is a serine/threonine kinase that phosphorylates cofilin and plays a critical role in integrin-mediated actin cytoskeletal reorganization and cell spreading. We previously showed that TESK1 interacts with Sprouty-4 (referred to as Spry4), an inhibitor of growth factor-induced Ras/MAP (mitogen-activated protein) kinase signalling, but the functional role of this interaction has remained unknown. In the present study, we show that Spry4 inhibits the kinase activity of TESK1 by binding to it through the C-terminal cysteine-rich region. Expression of Spry4 in cultured cells suppressed integrin-mediated cell spreading, and TESK1 reversed the inhibitory effect of Spry4 on cell spreading. Furthermore, Spry4 suppressed integrin- and TESK1-mediated cofilin phosphorylation during the spreading of cells on laminin. These findings suggest that Spry4 suppresses cell spreading by inhibiting the kinase activity of TESK1. Although tyrosine phosphorylation is required for the inhibitory activity of Spry4 on a Ras/MAP kinase pathway, mutation of the corresponding tyrosine residue (Tyr-75 in human Spry4) to an alanine had no apparent effect on its inhibitory actions on TESK1 activity and cell spreading, which suggests a novel cellular function of Spry to regulate the actin cytoskeleton, independent of its inhibitory activity on the Ras/MAP kinase signalling.

Animals↗
PubMed2002

Role of zyxin in differential cell spreading and proliferation of melanoma cells and melanocytes.

Cell spreading, proliferation, and survival are modulated by focal adhesions linking extracellular matrix proteins, integrins, and the cytoskeleton. Zyxin is a focal-adhesion-associated phosphoprotein with one domain involved in the control of actin assembly and three protein-protein adapter domains implicated in the regulation of cell growth and differentiation. We characterized zyxin expression in normal human melanocytes and six melanoma cell lines in relation to cell spreading, growth, and differentiation using Western immunoblotting techniques, image analysis, flow cytometry, and confocal microscopy. We found that zyxin, focal adhesion kinase, and paxillin were significantly upregulated in melanoma cells compared to melanocytes. Zyxin expression directly related to cell spreading and proliferation and inversely related to differentiation, whereas focal adhesion kinase correlated only to cell spreading and paxillin did not significantly correlate with any of the parameters. Treatment of melanoma cells with 12-O-tetradecanoylphorbol-13-acetate downregulated zyxin expression, inhibited cell spreading and proliferation, and promoted differentiation. In contrast, 12-O-tetradecanoylphorbol-13-acetate, a mitogen for melanocytes, induced upregulation of zyxin expression in melanocytes. These findings are consistent with a role of zyxin in modulation of cell spreading, proliferation, and differentiation. Therapies directed at the downregulation of this focal adhesion phosphoprotein in melanoma cells implicate a new approach for controlling melanoma cell growth.

Cell Differentiation↗