Fibronectin-independent adhesion of fibroblasts to extracellular matrix material: partial characterization of the matrix components

1983 ◽  
Vol 63 (1) ◽  
pp. 287-301
Author(s):  
P.A. Harper ◽  
P. Brown ◽  
R.L. Juliano

Fibroblasts can adhere to extracellular matrix (ECM) material by fibronectin-dependent (type I) and fibronectin-independent (type II) mechanisms. In this report we investigate the biochemical characteristics of ECM that contribute to type II adhesion. ECM capable of mediating type II adhesions is produced primarily by normal diploid fibroblasts, but not by transformed cells or epithelial cells. Treatment of fibroblast ECM under conditions that result in the removal of most of the ECM lipid or most of the ECM glycosaminoglycan does not impair type II adhesion. Likewise, treatment of the ECM with large amounts of purified collagenase does not block type II adhesion. However, treatment of ECM with low doses of trypsin or with an agent that reacts with tyrosine residues, results in complete ablation of the ability of the ECM to support type II adhesion. On the basis of these observations we suggest that the matrix component(s) mediating type II adhesion are non-collagenous proteins or glycoproteins.

1981 ◽  
Vol 91 (3) ◽  
pp. 647-653 ◽  
Author(s):  
P A Harper ◽  
R L Juliano

Fibroblastic CHO cells readily adhere to fibronectin (Fn) coated substrata. From the parental cell population we have recently selected a series of adhesion variants (ADV cells) that cannot adhere to Fn substrata (Harper and Juliano. 1980. J. Cell. Biol. 87:755-763). However, ADV cells readily adhere to substrata coated with extracellular matrix material (ECM) derived from human diploid fibroblasts by a mechanism that does not involve fibronectin (Harper and Juliano. 1981. Nature (Lond.). 290:136-138). Te Fn-dependent adhesion mechanism of parental cells (type 1 adhesion) and the ECM-dependent adhesion of ADV cells (type II adhesion) can also be discriminated on the basis of their differential sensitivity to proteolysis, with the type II mechanism being far more sensitive. In this communication we report that parental CHO cells possess both type I and type II mechanisms whereas ADV cells possess only the type II mechanism. We also identify a high molecular weight membrane glycoprotein (gp 265) that seems to play a role in type II adhesion. This component is detected by [125I]lactoperoxidase of [3H]borohydride-galactose oxidase labeling of surface proteins in WT and AD cells. Cleavage of gp 265 with low doses of proteases correlates completely with the loss of type II adhesion capacity. Thus CHO cells possess two functionally and biochemically distinct adhesion mechanisms, one involving exogenous Fn and the other mediated by the membrane component gp 265.


Author(s):  
Tugce Yuksel ◽  
Yalcin Yuksel ◽  
Busra Basaran ◽  
Esin Cevik

Block type quay walls are widely used as port structures in the world. In this study three types of vertical block type quay walls with different block size exposed to seismic loading were investigated experimentally. The block ratios of Type I, Type II and III vertical wall models are B/h=2, 1.5 and 1.5 & H/h=6, 6 and 3, respectively. The tests were conducted in the shaking tank with different harmonic seismic loadings and the behaviors of these walls were investigated comparatively.


1993 ◽  
Vol 265 (3) ◽  
pp. L250-L259
Author(s):  
E. H. Webster ◽  
S. R. Hilfer ◽  
R. L. Searls ◽  
J. Kornilow

The mesodermal capsule of the fetal lung plays a role in differentiation of the respiratory region. It has been proposed for other epithelial organs that the mesodermal capsule influences development by modifying the basal lamina or the extended extracellular matrix. The effect could be on deposition or turnover of collagens, proteoglycans, and/or glycoproteins. This study tests the role of glycoproteins in differentiation of respiratory endings by inhibiting their synthesis with the antibiotic tunicamycin (TM). Lungs at 16 and 18 days gestation and 3 days after birth were cultured with TM and examined for morphological and biochemical differences from normal controls. With TM, alveolar regions did not expand properly and formed fewer type I pneumocytes, although type II pneumocytes were unaffected. The epithelium of untreated respiratory regions showed greater incorporation of radioactive mannose than the airways region or mesenchyme. This incorporation was diminished in TM, but the pattern persisted. Comparison with the results obtained with beta-xyloside suggested that differentiation of type I and type II pneumocytes is under separate control.


1988 ◽  
Vol 34 (10) ◽  
pp. 2053-2057 ◽  
Author(s):  
S Raam ◽  
D M Vrabel

Abstract We present evidence to show that monoclonal antibodies to estrogen receptors (ER) in solid phase recognize the secondary estrogen binding sites with moderate to low affinity for estradiol (E2). An excellent quantitative agreement was found in five cytosols between the ER values obtained by the enzyme immunoassay (ER-EIA) and the amount of secondary estrogen binding sites measured by the assay involving dextran-coated charcoal (Clin Chem 1986;32:1496). The immunoreactive protein recognized by the antibody-coated beads, when allowed to react with ER(+) cytosols, is shown to bind [3H]estradiol only when the ligand concentration exceeds 8 nmol/L. Further biochemical and functional characterization of the immunoreactive protein is required to establish similarities/dissimilarities between this protein, high-affinity Type I ER sites, and the secondary sites such as Type II sites.


Materials ◽  
2003 ◽  
Author(s):  
Michael H. Santare ◽  
Wenzhong Tang ◽  
John E. Novotny ◽  
Suresh G. Advani

High-density polyethylene (HDPE) was used as the matrix material for a carbon nanotube (CNT) polymer composites. Multi-wall carbon nanotube composite films were fabricated using the melt processing method. Composite samples with 0%, 1%, 3% and 5% nanotube content by weight were tested. The mechanical properties of the films were measured by the small punch test and wear resistance was measured with a block-on-ring wear tester. Results show increases in the stiffness, peak load, work-to-failure and wear resistance with increasing nanotube content.


2020 ◽  
Vol 123 (4) ◽  
pp. 587-596
Author(s):  
A. Emanuel ◽  
C.H. Kasanzu ◽  
M. Kagya

Abstract Triassic to mid-Jurassic core samples of the Mandawa basin, southern Tanzania (western coast of the Indian Ocean), were geochemically analyzed in order to constrain source rock potentials and petroleum generation prospects of different stratigraphic formations within the coastal basin complex. The samples were collected from the Mihambia, Mbuo and Nondwa Formations in the basin. Geochemical characterization of source rocks intersected in exploration wells drilled between 503 to 4042 m below surface yielded highly variable organic matter contents (TOC) rated between fair and very good potential source rocks (0.5 to 8.7 wt%; mean ca. 2.3 wt%). Based on bulk geochemical data obtained in this study, the Mandawa source rocks are mainly Type I, Type II, Type III, mixed Types II/III and Type IV kerogens, with a predominance of Type II, Type III and mixed Type II/III. Based on pyrolysis data (Tmax 417 to 473oC; PI = 0.02 to 0.47; highly variable HI = 13 to 1 000 mg/gTOC; OI = 16 to 225 mg/g; and VR values of between 0.24 to 0.95% Ro) we suggest that the Triassic Mbuo Formation and possibly the mid-Jurassic Mihambia Formation have a higher potential for hydrocarbon generation than the Nondwa Formation as they are relatively thermally mature.


1996 ◽  
Vol 270 (6) ◽  
pp. L1017-L1022 ◽  
Author(s):  
I. Y. Adamson ◽  
L. Young

Most of the alveolar epithelium overlies a fused basement membrane produced by epithelial and endothelial cells. To determine how this type of matrix influences type II cell growth and function, we studied the effects of culturing isolated rat alveolar type II cells on an extracellular matrix (ECM) freshly produced by pulmonary vascular endothelial cells grown 5 days in culture. Type II cells from the same rats were cultured on plastic or Matrigel for comparison. A large increase in mitotic activity was seen in type II cells grown on the endothelial ECM at 2 days only; thereafter cells spread rapidly to confluence and lost their lamellar bodies. Cells grown on Matrigel remained cuboidal with lamellar bodies but grew more slowly, as judged by [3H]thymidine uptake and cell numbers. Incorporation of labeled choline into disaturated phosphatidylcholine (DSPC) was used as a marker of surfactant synthesis. After the rapid, brief burst of proliferation, type II cells on endothelial ECM showed a sudden decline in DSPC-DNA by day 4 compared with cells grown on matrigel. Binding of the lectin Bauhinia purpurea (BPA) indicated that after a phase of division, cells on endothelial ECM developed as type I epithelium by 4 days of culture, when > 70% of cells stained positively for BPA binding, whereas few cuboidal cells on Matrigel were stained. The results indicate that type II cells respond briefly to growth factors in pulmonary endothelial ECM; then this type of matrix promotes cell spreading with loss of type II function as cells subsequently resemble type I epithelium.


1989 ◽  
Vol 109 (3) ◽  
pp. 1363-1369 ◽  
Author(s):  
B Swoboda ◽  
R Holmdahl ◽  
H Stöss ◽  
K von der Mark

Collagen type XI is a component of hyaline cartilage consisting of alpha 1(XI), alpha 2(XI), and alpha 3(XI) chains; with 5-10% of the total collagen content, it is a minor but significant component next to type II collagen, but its function and precise localization in cartilaginous tissues is still unclear. Owing to the homology of the alpha 3(XI) and alpha 1(II) collagen chains, attempts to prepare specific antibodies to native type XI collagen have been unsuccessful in the past. In this study, we report on the preparation and use for immunohistochemistry of a polyclonal antibody specific for alpha 2(XI) denatured collagen chains. The antibody was prepared by immunization with the isolated alpha 2(XI) chain and reacts neither with native type XI collagen nor type I, II, V, or IX by ELISA or immunoblotting, nor with alpha 1(XI) or alpha 3(XI), but with alpha 2(XI) chains. Using this antibody, it was possible to specifically localize alpha 2(XI) in cartilage by pretreating tissue sections with 6 M urea. In double immunofluorescence staining experiments, the distribution of alpha 2(XI) as indicative for type XI collagen in fetal bovine and human cartilage was compared with that of type II collagen, using a monoclonal antibody to alpha 1(II). Type XI collagen was found throughout the matrix of hyaline cartilage. However, owing to cross-reactivity of the monoclonal anti-alpha 1(II) with alpha 3(XI), both antibodies produced the same staining pattern. Cellular heterogeneity was, however, detected in monolayer cultures of human chondrocytes.(ABSTRACT TRUNCATED AT 250 WORDS)


1999 ◽  
Vol 589 ◽  
Author(s):  
W. Mader ◽  
B. Freitag ◽  
K. Kelm ◽  
R. Telle ◽  
C. Schmalzried

AbstractThe structure and chemical composition of two types of precipitates in the system TiB2-WB2-CrB2 were studied by means of high-resolution TEM and energy filtering TEM. Type I particles (W2B5 structure) are precipitated at the basal plane of the hexagonal matrix whereas type II precipitates are thin platelets lying parallel to the {1100} prism planes. Lattice imaging yields displacements of the metal positions with respect to the matrix. Information on the chemical composition at high lateral resolution is obtained from elemental maps of all chemical constituents using electron spectroscopic imaging (ESI). The type II precipitates show a decrease in the B and Ti concentration, whereas the tungsten concentration increases and the Cr is homogeneously distributed. The HRTEM results combined with the results of the elemental maps allow to develop a structural model based on the intergrowth of the β-WB structure in the TiB2-rich matrix. The two deficient boron layers in W0.5Ti0.5B with a spacing of 0.38 nm can be used to examine the resolution limit of ESI.


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