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C Guidry

Publications and source records attributed to C Guidry.

26 records · Page 2Linked to original sources

Endothelial cells secrete a factor that promotes fibroblast contraction of hydrated collagen gels.

Bovine aortic endothelial cells (BAEC), grown in vitro, are shown to synthesize and secrete factor(s) that stimulate fibroblasts to contract collagen matrices. The amount of contraction-promoting activity in the conditioned media is dependent on conditioning time and the number of cells in the culture. Production of the contraction-promoting activity continues at a high stable level for at least 5 d in serum-free medium but is abolished when the cells are exposed to an inhibitor of protein synthesis. The mechanism of action of the contraction factor(s) derived from endothelial cells was compared with that of unidentified serum factors. The endothelial cell-secreted factor(s) depends on active protein synthesis by the target cell but does not need to be present during the contraction process. The serum factors on the other hand promote collagen contraction in the absence of de novo protein synthesis but need to be continuously present. Preliminary biochemical characterization of the contraction-promoting factors produced by endothelial cells revealed properties similar to those of previously identified growth factors. However, the BAEC-secreted factor was found to be distinct from a previously identified contraction-promoting transforming growth factor beta.

Animals↗

Contraction of hydrated collagen gels by fibroblasts: evidence for two mechanisms by which collagen fibrils are stabilized.

Studies were conducted to learn more about the mechanism by which fibroblasts contract hydrated collagen gels, a process that may be important in the supramolecular organization of the extracellular matrix. Removal of cells from contracted gels by two different methods, treatment with detergent or treatment with trypsin/EDTA solution, had no visible effect on the bundles of collagen fibrils that had been organized in frameworks around and in between the cells. There was, however, a portion of the collagen gels that expanded after the cells were removed. We conclude that during concentration of collagen gels by fibroblasts, rearranged collagen fibrils were stabilized in place by two different mechanisms. At first, the fibrils were mechanically held in place by the cells. Subsequently, the fibrils were stabilized by non-covalent chemical interactions that are independent of cells. A model system for studying collagen gel reorganization in the absence of cells was developed based on centrifugation of the gels. The overall features of collagen gel reorganization by centrifugation were similar to the features of collagen gel contraction by cells. The collagen fibrils of gels reorganized by centrifugation were at first stabilized mechanically and the gels expanded after centrifugation was stopped. With additional time, the collagen fibrils were stabilized by non-covalent chemical interactions, and then the gels no longer expanded after centrifugation was stopped.

Cells, Cultured↗

Heparin modulates the organization of hydrated collagen gels and inhibits gel contraction by fibroblasts.

We studied the effects of extracellular matrix components on fibroblast contraction of hydrated collagen gels. After 4-h incubations, heparin-containing collagen gels contracted only 10% compared with 50% contraction of control gels. Contraction was not affected by hyaluronic acid, dermatan sulfate, or fibronectin, implying that the activity of heparin was specific. The possibility that heparin inhibited attachment of the cells to the gels was ruled out. Also, addition of heparin to the incubation medium had no effect on contraction. Microscopic examination showed that control collagen gels were composed of a uniform network of interlocking fibrils of similar sizes. Heparin-containing gels, on the other hand, were highly variable with some collagen bundles containing 5-6 collagen fibrils and other regions containing amorphous material. Unlike the control gels, the fibrils of heparin-containing gels were not continuously interconnected. Based on the results, we propose that fibroblasts attach normally to the collagen fibrils of heparin-containing gels and attempt to contract the gels, but the mechanical forces exerted by fibroblasts on individual collagen fibrils cannot be propagated throughout the gels.

Cells, Cultured↗

Studies on the mechanism of hydrated collagen gel reorganization by human skin fibroblasts.

During reorganization of collagen gels by human skin fibroblasts the total protein content of the gels remained approximately constant. Only 5% of the collagen was degraded, although the volume of the gels decreased by 85% or more. It could be concluded, therefore, that gel reorganization required physical rearrangement of pre-existing collagen fibrils rather than degradation of the original collagen and resynthesis of a new matrix. Collagen molecules in the gels were not covalently crosslinked or otherwise modified enzymically during gel reorganization, as determined by sodium dodecyl sulphate/polyacrylamide gel electrophoresis and collagen repolymerization studies. Serum was required for gel reorganization and, in the absence of serum, cell spreading was predominantly filipodial, i.e. there was little cytoplasmic reorganization. At the electron-microscopic level it was found that many more collagen fibrils became associated with the cells in the presence of serum than in its absence. Serum was also found to promote the synthesis and secretion of proteins by the cells, and conditioned medium could take the place of serum in promoting gel reorganization. The involvement of cell-secreted factors was also demonstrated by the ability of cycloheximide to inhibit gel reorganization. Finally, when gel reorganization was stopped by adding cytochalasin D to the incubations or removing cells by detergent treatment, a small but significant re-expansion of the collagen fibrils was observed. Consequently, a portion of the collagen that had been physically reorganized by the gels was unstable and could not hold its position without continued force exerted by the cells.

Cell Count↗

Diet, nutrition intake, and metabolism in populations at high and low risk for colon cancer. Metabolism of neutral sterols.

Cholesterol and its metabolites, together with bile acids, are implicated as risk factors in the genesis and progression of colon cancer. This study was designed to determine differences in the neutral sterol composition of stools from four different population groups differing in their dietary habits as well as in their expected rates for colon cancer. Four study groups consisting of 18 Seventh-day Adventist (SDA) pure vegetarians, 50 SDA lacto-ovo vegetarians, 50 SDA nonvegetarians, and 50 general population nonvegetarians were selected from the greater Los Angeles Basin area. Three-day composite stool samples were lyophilized and then analyzed for their neutral sterol composition. Cholesterol excretion values consistently showed an age-dependent peak in 46- to 50-yr age group for the total population, SDA lacto-ovo vegetarian and SDA-nonvegetarian subgroups being the principal contributors to this age-dependent phenomenon. The SDA pure vegetarians exhibited the lowest fecal concentrations and daily excretion of cholesterol as expected since their intake of dietary cholesterol is insignificant. Among the other SDA, regardless of whether they are lacto-ovo vegetarians or nonvegetarians, their cholesterol excretion patterns were similar but higher than in the nonvegetarians from the general population. Since dietary intakes of cholesterol are not significantly different among the two nonvegetarian groups, the differences in excretion values are attributable to differences in colonic metabolism. The ratio of cholesterol/cholesterol metabolites showed generally lower values among nonvegetarians compared to the matched group of lacto-ovo vegetarians. The observation was made that fecal cholesterol and its metabolites tend to be higher among nonvegetarians compared to those in the corresponding vegetarian groups.

Adult↗

Fecal bile acids and neutral sterols in the cotton-top tamarin (Saguinus oedipus).

1. The cotton-top tamarin (Saguinus oedipus), a small New World primate susceptible to spontaneous development of colon cancer, was studied for its fecal neutral sterol and bile salt composition. 2. Standardization procedures to establish the effect of exposure of the stool to room temperature air for various time-periods showed no significant effects on the neutral sterol and bile salt composition of the samples. 3. Microbial degradation of cholesterol and bile acids to secondary metabolites showed a progressive rise during the first year of life after which some degree of homeostasis was observed. 4. The proportion of cholesterol that remained unmetabolized by colonic microflora was in excess of 50%, an amount that was significantly higher than in man and other higher primates. 5. Ursodeoxycholic acid was identified as a significant (12%) component of fecal bile acids in this species. 6. Secondary bile acids formed by the action of enteric microflora were also significantly lower than levels found in man and other animals.

Animals↗