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Biomedical subjects

L Zardi

Publications and source records attributed to L Zardi.

At least 145 records · Page 8Linked to original sources
PubMed1984

Characterization of synovial fluid fibronectin from patients with rheumatic inflammatory diseases and healthy subjects.

Synovial fluid fibronectin from normal subjects and from patients who have rheumatic inflammatory diseases has been studied and compared with plasma fibronectin. The average fibronectin concentration in synovial fluids from normal subjects was 172 +/- 69 micrograms/ml; it was 721 +/- 315 and 556 +/- 349 micrograms/ml in synovial fluids from patients with rheumatoid arthritis and osteoarthritis, respectively. This is the first report on fibronectin concentrations in normal synovial fluids. Synovial fluid fibronectin from healthy subjects and patients with rheumatoid arthritis or osteoarthritis showed a molecular weight identical to that of plasma fibronectin. All normal and pathologic synovial fluid fibronectins showed a remarkably lower electrophoretic mobility compared with that of plasma fibronectin, when separated according to net molecular charge on agarose gel. Peptides from thermolysin digests of fibronectin from plasma and synovial fluid, when compared on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, showed distinct differences. These data demonstrate that synovial fluid fibronectin represents a molecular form which is structurally different from that of plasma fibronectin. This suggests that synovial fluid fibronectin is locally synthesized, possibly by a cell type which differs from that responsible for the production of the plasmatic fibronectin pool.

Aged↗
PubMed1984

Fibronectin concentrations in pleural effusions of patients with malignant and non-malignant diseases.

We have determined the amount of fibronectin in pleural fluids and plasma of 131 patients suffering from pleurisy of malignant and non-malignant etiology. We observed a significantly higher concentration of fibronectin in pleural fluids of a few patients with mesothelioma as compared with that found in pleural fluids of patients with pleurisies of different etiology. In these patients the pleural fluid fibronectin concentration was higher than that measured in plasma, suggesting local synthesis of fibronectin.

Adult↗
PubMed1983

Differences in domain structure between pericellular matrix and plasma fibronectins as revealed by domain-specific antibodies combined with limited proteolysis and S-cyanylation: a preliminary note.

Differences in domain structure between human fibronectins obtained from pericellular matrix and plasma have been revealed by limited proteolysis and S-cyanylation, followed by identification of each domain with domain-specific antibodies. Although the overall domain structure is similar between pericellular and plasma fibronectins, the fragments derived from the COOH-terminal region of these fibronectins, which were defined by specific antibodies, exhibited clear differences in their molecular weights and protease susceptibility, suggesting that the structure near the COOH-terminal region is significantly different between these two proteins.

Antibodies↗
PubMed1983

Increased plasma fibronectin concentrations in tumor bearing mice.

Fibronectin concentration was studied in plasma of mice at different times after inoculation of 3 different experimental tumors, Colon 26 adenocarcinoma, mFS6 fibrosarcoma and B16 melanoma. A significant increase in fibronectin concentrations 10-30 days after inoculation of all tumors was found. Highest and earliest fibronectin increases were observed in BALB/c mice inoculated with Colon 26 adenocarcinoma cells. Control animals inoculated with previously irradiated cells (5000 rad) showed neither tumor growth nor significant variations in plasma fibronectin levels.

Adenocarcinoma↗
PubMed1983

Can heterologous collagen enhance the granulation tissue growth? An experimental study.

A purified and lyophilized sponge of native bovine tendon collagen (Gelfix) was used to plug gastric, jejunal and colonic surgical holes (phi = 6 mm) on 78 white rats. In a control group (18 rats) colonic holes were plugged with autologous epiploon. The mortality rate due to generalized peritonitis, was higher (Fischer test = 4%) in the control group than in animals treated with the heterologous collagen. The healing process was evaluated by light microscopy at sequential intervals ranging from 4 to 120 days after implantation. The heterologous collagen biological activity was evaluated as follows: 1. The sponge was labelled with Tyrosine 125I or mixed with Collagen Methyl 14C methylated. Then the radioactivity of the isolated granulation tissue cells was evaluated. The electrophoresis of these lysed cells, as well as histoautoradiography were performed. 2. Indirect immunofluorescence studies employing rabbit antibodies against fibronectin were also carried out. The heterologous collagen seemed to act as a mechanical support and as a stimulus to chemotactic migration of fibroblasts. It also appeared to participate as a nutrient substitute in the metabolic activity of the granulation tissue. Finally, heterologous collagen fibers were found to be massively bound by fibronectin.

Animals↗
PubMed1982

Monoclonal antibodies in analysis of cathepsin G-digested proteolytic fragments of human plasma fibronectin.

Proteolytic fragments of fibronectin, obtained by digestion with cathepsin G, were transferred electrophoretically from sodium dodecyl sulphate (NaDodSO4) polyacrylamide gels to nitrocellulose sheets and used as antigens for monoclonal antibodies. All 9 monoclonal antibodies tested reacted with undenatured intact fibronectin or its fragments applied directly to nitrocellulose sheets. Two of the clones did not react with the NaDodSO4-treated transferred material suggesting reactivity with conformational determinants. Distinct fragments of fibronectin could be detected by several of the antibodies. None of the monoclonal or the polyclonal antibodies used reacted with the Mr = 40,000 or Mr = 30,000 gelatin-binding fragments of fibronectin. However, one of the monoclonal antibodies reacted specifically with their precursor Mr = 64,000 fragment, but apparently with its gelatin-nonbinding segment. The apparent non-immunogenicity of the gelatin-binding domain is conspicuous, suggesting that it may be highly conserved in evolution. The present method, combination of controlled proteolytic cleavage with electrophoretic transfer, provides an effective means for characterization of monoclonal antibodies raised against proteins.

Animals↗
PubMed1982

Species-specific monoclonal antibodies in the assignment of the gene for human fibronectin to chromosome 2.

Using three different species-specific monoclonal antibodies we have studied, in human-mouse and human-hamster somatic cell hybrids, the correlation between the presence of different human chromosomes and the ability to release human fibronectin into the tissue culture medium. Presence of human fibronectin was determined by an affinity-radioimmunoassay. In addition, tissue culture media of the different hybrids were separated on SDS-polyacrylamide gels, the proteins were blotted onto a nitrocellulose sheet and human fibronectin visualized by an immunoenzymatic technique. Karyology and determination of isoenzyme markers of specific human chromosomes show that the ability to produce human fibronectin segregated with the presence of human chromosome 2.

Animals↗
PubMed1980

Somatic cell hybrids producing antibodies specific to human fibronectin.

We have produced somatic cell hybrids between mouse plasocytoma cells deficient in hypoxanthine phosphoribosyltransferase (P3 x 63 Ag8) and spleen cells derived from mice immunized with purified human plasma fibronectin. We report herein the properties of monoclonal anti-fibronectin antibodies released by eight different clones.

Animals↗
PubMed1980

A simplified procedure for the preparation of antibodies to serum fibronectin.

In the present paper we describe in detail a simple procedure for the preparation of monospecific antisera to human and mouse serum fibronectin. A similar procedure could also be used to prepare antibodies to fibronectin from other species. The procedure, based on the recently reported affinity of fibronectin for gelatin, essentially consists of two steps (1) Immunization of rabbits with fibronectin purified from serum by affinity chromatography using gelatin coupled to CNBr-activated Sepharose 4B. (2) Absorption of the antiserum obtained by an immunoabsorbent prepared using fibronectin-free serum proteins that remained after absorbing serum with gelatin-Sepharose. The antisera obtained were monospecific, as determined by immunoelectrophoresis and did not show any difference with respect to antisera prepared by different procedures.

Animals↗
PubMed1979

Fibronectin: a chromatin-associated protein?

We have previously reported that chromatin preparations from human cultured fibroblasts contain a single homologous serum protein. In this paper we present evidence, based on immunological identity and physicochemical properties, that this serum protein is fibronectin. Furthermore, using a radioimmunoassay system, we have estimated that fibronectin represents about 0.7% of the total protein in both chromatin preparations and whole fibroblasts. Using a nitrocellulose filter assay system, we also show that fibronectin is a DNA-binding protein having an equilibrium constant of 4.6 x 10(-6) M. Equilibrium competition experiments have demonstrated that fibronectin has the ability to differentiate among nucleotides, indicating that fibronectin-DNA interaction is at least partially specific, and that a minimum polymer length of 12-18 nucleotides is required for effective binding to occur. Fibronectin has been isolated readily from plasma using DNA-affinity chromatography. We do not have direct evidence that fibronectin is an actual nonhistone chromosomal protein, but fibronectin is a DNA-binding protein (at least under in vitro assay conditions) and appears to be a normal constituent of chromatin as chromatin is currently isolated from cell nuclei.

Antibody Specificity↗
PubMed1979

Concentration of fibronectin in plasma of tumor-bearing mice and synthesis by Ehrlich ascites tumor cells.

In the present paper we have studied: (a) the concentration of fibronectin (FN) in plasma and in ascitic fluid of mice at different times after inoculation of Ehrlich ascites tumor cells; (b) the ability of Ehrlich ascites cells to synthesize and release FN; and (c) the localization of FN in Ehrlich ascites cells by immunofluorescence microscopy. It was found that (a) 4 to 5 days after inoculation of the tumor, the plasma concentration of FN was significantly higher [1.7 +/- 0.07% (S.E.) of total plasma protein] than that in the normal control mice (0.8 +/- 0.035); (b) FN is present in the ascitic fluid in all phases of tumor growth; (c) Ehrlich ascites cells cultured in vitro synthesize and release large amounts of FN in the culture medium; and (d) only about 1 to 2% of the tumor cells show a very small amount of FN, and this is mostly in the area of cell-cell contact.

Animals↗
PubMed1977

Effect of split doses of N-methyl-N-nitrosourea on DNA repair synthesis in cultured mammalian cells.

DNA repair was measured in human fibroblasts, mouse C3H 10 T 1/2 fibroblsts and rat hepatocytes by the non-semi-conservative incorporation of [3H]-TdR during DNA repair synthesis using liquid scintillation techniques. Confluent monolayers of these cells grown on cover slips were exposed to split doses (125 or 250 microgram/ml) of the mutagenic and carcinogenic alkylating agent MNU and DNA repair synthesis compared with that produced by a single dose (500 microgram/ml). No significant difference in DNA repair capacity was detected in the three cell lines treated with a single dose or split doses of MNU.

Cells, Cultured↗