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Biomedical subjects

F Pezzetti

Publications and source records attributed to F Pezzetti.

At least 55 records · Page 3Linked to original sources
PubMed1994

Chondroitin sulphates and embryonic chick skin fibroblast proliferation.

We have investigated the action of sulphated glycosaminoglycans (GAGs) upon primary 11-day chick embryo skin fibroblasts cultured for various periods of time in the presence or absence of fetal calf serum (FCS). Chondroitin 4- and 6-sulphate (CS) added to serum supplemented medium provoked a decrease in DNA synthesis both in sparse and crowded cultures only at high concentrations (250 micrograms/ml). Synthesis of endocellular and secreted proteins was not affected. In contrast, CS administered to fibroblasts for 24 h in serum-free medium stimulated DNA synthesis. We postulate that the CS stimulating effect seen in serum-free cultures might be antagonized by peptide regulators of cell growth normally present in serum.

Animals↗
PubMed1994

Interleukin-1 and interleukin-6 differentially regulate the accumulation of newly synthesized extracellular matrix components and the cytokine release by developing chick embryo skin fibroblasts.

In the present study, we demonstrate that both interleukin-1 (IL-1) and interleukin-6 (IL-6) induced a significant decrease in glycosaminoglycan (GAG) synthesis and, more strikingly, secretion by 7 and 13 day-old chick embryo skin fibroblasts. We demonstrated that interleukin treatment also inhibited the synthesis of collagenase-digestible proteins (type I collagen). In addition, tissue culture supernatants (conditioned media, CM) were tested for reactivity for IL specific ELISAs and for their ability to stimulate proliferative responses in mouse thymocytes and hybridoma cells. Our findings demonstrate that chick embryo skin fibroblasts spontaneously produce IL-1 and, in even greater amounts, IL-6. Highest levels of interleukin secretion were found in the CM of 13 day-old fibroblasts and the IL-1 beta isoform was predominant over IL-1 alpha. Pretreatment of the fibroblasts with either IL-1 or IL-6 increased the secretion of both cytokines. Increased IL-1 levels were correlated with enhanced IL-1 bioactivity in the CM of IL-6 treated fibroblasts. By contrast, the raised concentrations of IL-1 in the CM of IL-1 treated cells and IL-6 in the CM of IL-1 or IL-6 treated fibroblasts failed to translate into augmented bioactivity. These observations, taken together, indicated that IL-1 and IL-6 are able to regulate the synthesis and secretion of ECM macromolecules of developing connective tissues and the cytokine release by chick embryo skin fibroblasts.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Age related effects of lectins on GAG metabolism in cultured embryonic fibroblasts.

Lectins (WGA, SBA or PNA) were administered to 7 and 14 days chick embryo fibroblasts to evaluate their ability to influence GAG cellular and extracellular accumulation. HA and sulphated GAG were differently affected by lectins. WGA addition to culture medium enhanced HA and sulphated GAG in the two examined compartments. SBA and PNA increased sulphated GAG only in the cellular pool with a consequent reduction of secreted GAG. As far as HA is concerned, SBA and PNA promoted its accumulation in the extracellular compartment. NH4Cl administration indicated a decline of GAG degradation in the course of development in presence of lectins. Generally the effect of lectins was greater in the youngest stage: showing that lectins were able to bind 7 and 14 days-old fibroblasts differently.

Aging↗
PubMed1993

Embryonic skin fibroblasts release TGF alpha and TGF beta able to influence synthesis and secretion of GAG.

Conditioned medium (CM), collected from 7 and 14 days-old chick embryo skin fibroblasts and added to the same cells, increases glycosaminoglycans (GAG) intra- and extracellular accumulation. The factors responsible for GAG enhancement are TGF alpha and TGF beta because they are trypsin and dithiothreitol sensitive, stable or enhanced by heat and transient acidification. Moreover, Sephadex G-75 fractions of CM active on GAG synthesis contain, when analysed on SDS-polyacrylamide gel electrophoresis, two bands that comigrate with TGF alpha and TGF beta and induce NRK cells clone 49F to form large colonies of mean size > 8.000 microns 2 in soft agar. Since both the factors must be present to induce the formation of large colonies we come to the conclusion that CM contains TGF alpha and TGF beta. The two growth factors have different effects on the accumulation of individual classes of GAG in the ECM. In particular, TGF beta stimulates a marked increase of CS and DS, TGF alpha of HA and DS in the medium. The contemporaneous addition of TGF alpha and TGF beta to 7 days-old fibroblasts produces a pattern of GAG response similar to CM. These embryonic fibroblasts may control their own GAG synthesis and secretion through autocrine TGF alpha and beta activity.

Animals↗
PubMed1993

Hyaluronic acid modulates growth, morphology and cytoskeleton in embryonic chick skin fibroblasts.

The action that hyaluronic acid (HA) exerts on cell proliferation was investigated in embryonic chick skin fibroblasts at different ages (7-14 days) and in different cell-cycle stages evaluated by flow cytometry (cells maintained with and without serum). Proliferation was estimated by 3H-thymidine incorporation and cell counting. The results demonstrated hyaluronic acid inhibits cell multiplication in all different environmental conditions examined. The inhibitory effect of HA is more evident in 14-day than 7-day old fibroblasts. The ability of HA to modulate 3H-thymidine incorporation did not involve a change in the time required for cells entering the S phase of the replicating cycle, but is due to a smaller number of cells entering in this phase. As the relationships between components of the extracellular matrix (ECM) and the cytoskeleton are known, parallel studies were carried out on some cytoskeleton proteins. Furthermore, by modifying the capacity of cells to adhere to the substrate, HA induced alterations in cell shape and in cytoskeleton components involved in these processes. We may hypothesize that HA, binding specific membrane receptors, affects cell adhesion and morphology inducing less receptivity of fibroblasts to mitogenic stimuli by transmembrane interactions with cytoskeleton.

Actinin↗
PubMed1991

Analysis of the patterns of expression of mRNAs for the alpha- and beta-subunits of the lysosomal enzyme beta-N-acetylhexosaminidase in mouse epididymis and testis.

mRNA for the alpha- and beta-subunits of mouse beta-N-acetylhexosaminidase were detected in tissue sections of epididymis and testis using 3H-labelled antisense RNA probes by hybridization in situ. There were clear differences in the intensity of the signals given by the two tissues with alpha-subunit mRNA being more abundant in testis than epididymis, while the opposite was so with beta-subunit mRNA. Differences in the isoenzyme patterns of the two tissues are likely to be due, at least in part, to differences in transcription by the Hexa and Hexb genes.

Animals↗
PubMed1991

beta-Hexosaminidase expression in chick embryo fibroblasts in vitro.

1. Two forms of beta-hexosaminidase, similar to hexosaminidase A and hexosaminidase C, were separated by DEAE-cellulose chromatography in chick embryo skin fibroblasts in vitro. 2. beta-Hexosaminidase specific activity increases during development in cultured chick embryo skin fibroblasts in vitro. 3. Concanavalin-A treatment determines the increase of the neutral form, hexosaminidase C, during development. 4. Concanavalin-A reduces the specific activity of beta-hexosaminidase during development.

Animals↗
PubMed1991

Effects of steroids on human normal and otosclerotic osteoblastic cells: influence on thymidine and leucine uptake and incorporation.

Steroid hormones are able to influence the metabolism of bone tissue in vivo, but reports regarding their direct action on bone cells fail to agree. In this study, in vitro administration of 17 beta-estradiol, testosterone and corticosterone to normal and otosclerotic osteoblastic cells induced a drop in DNA synthesis in both populations, an increase in the neosynthesis of endocellular proteins in normal cells and a rise, mainly in proteins secreted into the medium, in otosclerotic cells. The fact that 3H-thymidine and 3H-leucine uptake were lower in otosclerotic than in normal cells suggests that the membrane permeability differs in the two populations and that steroids exert an influence on both isotope uptake and directly modulate DNA and protein synthesis.

Corticosterone↗
PubMed1990

Cytoskeletal and DNA synthesis modification by concanavalin A in embryonic fibroblasts maintained in serum-free and serum-added medium.

The administration of lectin concanavalin A (Con A) to in vitro cultures of chick embryo fibroblasts caused dose-dependent changes in cell shape, cytoskeleton network and DNA synthesis. After 6 hrs. even a low doses of Con A produced evident effects in serum-free cultures, whereas higher doses were required to cause alterations in cells cultured in serum-added medium. As the sugar competitor mannoside abolishes the effects, it would seem that the lectin acts by binding to transmembrane receptors and that the fibronectin present in the serum engages the receptors so that they are not available to Con A.

Animals↗
PubMed1990

Restoration of a normal phenotype, microtubular pattern and DNA synthesis in embryonic fibroblasts concanavalin A pretreated.

The present research investigated the time required for reconstructing normal microtubular pattern and the phenotype of cells after short-term (30 min.) and long-term (24 hrs.) pretreatment with the lectin concanavalin A (Con A). Short-term pretreatment led to the formation of incomplete tubules within 2 and 24 hrs. in cells cultured in 199 medium alone. The addition of serum to the medium reversed the globular phenotype and allowed the formation of normal microtubules, even after prolonged pretreatment with Con A. Whereas long-term Con A treatment provoked a reduction in DNA synthesis in 199 alone, in serum-added 199 the percentage of 3H-thymidine incorporation of treated cells tended to reach that of controls over time.

Animals↗
PubMed1989

beta-N-acetylhexosaminidase isoenzymes during chick embryo development.

1. Two forms (I and II) with acidic pH optima and a neutral form of beta-hexosaminidase has been separated by DEAE-cellulose chromatography and characterized in skin and lung of 7, 9, 11, 14 day chick embryos and 1 day old chicken. 2. Forms I and II are similar to hexosaminidase A and B for their behaviour on DEAE-cellulose chromatography, Concanavalin A-Sepharose column and thermal stability. 3. Neutral form has a neutral pH optimum and higher molecular weight and a more acidic I. P. than forms I and II, a low beta-N-acetylgalactosaminidase activity and it is not bound by a Concanavalin A-Sepharose column and in that resemble hexosaminidase C and/or other neutral hexosaminidases. 4. We have found differences in the percentage of neutral form and in the specific activities of the extracts in the skin in different stages of development. 5. No significant differences were observed in the lung.

Animals↗
PubMed1989

Ability of retinoic and ascorbic acid to interfere with the binding of benzo(a)pyrene to DNA in explants from donors with bronchial cancer.

The capability of ascorbic acid (AA) and transretinoic acid (RA) to interfere with 3H-benzo(a)pyrene [B(a)P] binding to DNA has been evaluated in cultured bronchial mucosa explants from patients with bronchial cancer. The results show that the DNA-bound 3H-B(a)P is smaller in treated cultures than in controls. To explain this finding, it is proposed that AA, acting as antioxidant, inhibits the oxidative degradation of B(a)P, and that RA, a lipophilic compound interacting with the lipid components of mixed function oxidases, could modify the activities of these enzymes. Both vitamins decrease the concentration of ultimate carcinogen metabolites, which can interact with DNA. Furthermore, the treatment with RA does not increase DNA synthesis, while AA inhibits 3H-thymidine incorporation.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1989

Beta-N-acetyl-D-glucosaminidase activity in embryonic chick skin and lung tissue and cultured fibroblasts.

During development the content of mesenchymal glycosaminoglycans (GAG) undergoes prominent changes, currently considered to act as regulatory signals in the epithelial-mesenchymal interactions. The factors involved in controlling GAG composition are as yet completely unknown. Lysosomal enzymes play a key role in GAG turnover. A possible mechanism for regulating GAG content could therefore be linked to developmental modulation of lysosomal glycosidases activity. We have examined the activity of the beta-N-acetyl-D-glucosaminidase (EC 3.2.1.30; a lysosomal hydrolase cleaving glycosidic linkage of the non-reducing terminal beta-N-acetyl-D-glucosamine residues) in chick embryo skin and lung (rudiments whose GAG composition has previously been studied) at various embryonic stages. Determinations were carried out on whole organs as well as on primary cultures of fibroblasts obtained from the two rudiments. beta-N-acetyl-D-glucosaminidase activity varied greatly during development, and it was significantly different in embryonic skin and lung tissues at various incubation days. In cultured fibroblasts, the enzymatic activity varied at different incubation days correlating with the in vivo data. Developmental changes of beta-N-acetyl-D-glucosaminidase paralleled mesenchymal GAG pattern both in vivo and in vitro. Our results, therefore, support the possibility that lysosomal enzymes could be involved in the regulation of mesenchymal GAG content during development.

Acetylglucosaminidase↗
PubMed1988

Different response of chicken embryo fibroblasts and hepatocytes to the interference of certain antioxidants on the binding of [G-3H]benzo[a]pyrene to DNA.

The effects of treatment with reduced glutathione, cysteine and ascorbic acid on chick embryo fibroblasts and hepatocytes cultured in vitro in the presence of [G-3H]benzo[a]pyrene ([3H]BP) were studied in an attempt to evaluate the capacity of these antioxidants to interfere with the binding of the carcinogen to DNA in these two cell populations. The results revealed that in the absence of treatment with antioxidants, the carcinogen bound to the DNA of the hepatocytes less than it did to that of the fibroblasts, despite the fact that the two cell populations manifest more or less the same uptake of [G-3H]benzo[a]pyrene from the medium. Moreover the hepatocytes, unlike the fibroblasts, seemed to reap little advantage from the known capacity of the antioxidants to interfere with the binding of the carcinogen to DNA.

Animals↗
PubMed1988

Age related and lectin influenced changes of exoglycosidases activity in cultured chick embryonic skin fibroblasts.

beta-N-acetyl-D-glucosaminidase, beta-N-acetyl-galactosaminidase and a beta-D-galactosidase activity was determined in untreated and lectins (ConA, PNA, SBA and WGA) treated chick embryonic skin fibroblasts at two incubation stages. Activity of all three glycosidases increased between 7 and 14 incubation days. ConA and WGA affected the levels of enzymatic activity; while SBA and PNA were uneffective. We discuss these findings in relation to a possible role of glycosidases in controlling mesenchymal GAG turnover.

Acetylglucosaminidase↗
PubMed1986

Antagonism between catalase and ascorbic acid in control of normal and neoplastic cell multiplication.

The authors studied the effects of a treatment with ascorbic acid on in vitro multiplication of ascites tumour cells (ATP C+), of fibroblast-like cells and of hepatocytes from chick embryos, by measuring [3H]thymidine incorporation into DNA. The results obtained show that the ATP C+ cells are the most sensitive to the toxic effects of the experimental treatment, while the hepatocytes are the most resistant cell population. A treatment with catalase was able to greatly reduce the damage caused by ascorbic acid on the ATP C+ cells. It is hypothesized that ascorbic acid inhibits cell multiplication by the H2O2 formed by its oxidation and that the cells having the highest level of catalase are more resistant to its toxic effects.

Animals↗
PubMed1986

Inhibition of the binding of 7,12-dimethylbenz[a]anthracene to DNA by ascorbic acid, reduced glutathione and cysteine in chick embryo cells cultured in vitro.

A study on the capacity of ascorbic acid, reduced glutathione and cysteine to interfere with 3H-7,12-dimethylbenz[a]anthracene (3H-DMBA) binding to DNA in cultured fibroblast-like cells from 11-day-old chick embryos showed that, although the total amount of 3H-DMBA in the treated cells was greater than in the untreated cells, the DNA-bound 3H-DMBA was less. Comparisons between the various experimental groups demonstrated that the greater 3H-DMBA in the ascorbic acid-, reduced glutathione-, and cysteine-treated groups could not be attributed to an initially higher number of cells, nor to a treatment-induced increase in DNA synthesis. It is proposed that the three substances examined inhibit the oxidative degradation of 3H-DMBA, thereby favoring its accumulation within the cell and reducing the formation of DNA-binding metabolites.

9,10-Dimethyl-1,2-benzanthracene↗