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M Castagna

Publications and source records attributed to M Castagna.

At least 73 records · Page 4Linked to original sources
PubMed1988

Morphologic and antigenic properties of mouse mammary tumor virus produced in a hormone-responsive fashion by C57Bl/10 mammary tumors of non-viral origin.

Bl-MaTU/A1 mouse mammary tumor cells, derived from a C57B1/10 mammary adenocarcinoma induced by dimethylbenzanthracene and mammotropic hormones, express virus particles and proteins related to mouse mammary tumor virus (MMTV). Immunocytochemical analysis by means of monospecific and monoclonal anti-gp52 sera revealed a different localization of the main structural proteins of MMTV in Bl-MaTU/A1 and GR cells (the latter used as a positive virus-producing control). Immunoelectron microscopy of B-type particles budding from the microvilli of dexamethasone-stimulated Bl-MaTU/A1 cells showed remarkably weak reactivity of the viral envelope with anti-gp52-protein A-gold complexes as compared with that of dexamethasone-stimulated GR cells. Since Bl-MaTU/A1-associated MMTV originates from the amplified unit II of endogenous MMTV, which is altered probably within the env gene, the observed antigenic difference in the Bl-MaTU/A1-associated MMTV may be due to altered synthesis of gp52 glycoprotein in these cells.

Adenocarcinoma↗
PubMed1987

Alkyl analogs of diacylglycerol as activators of protein kinase C.

Diacylglycerols which activate protein kinase C have the 1,2-sn configuration. Short-chain saturated fatty acids or long-chain unsaturated fatty acids are required for supporting the potency of these lipids. Using alkyl analogs such as 1-O-decyl-2-O-decanoylglycerol, 1-O-decanoyl-2-O-decylglycerol, 1,2-O-didecylglycerol, 1-O-hexadecyl-2-O-acetylglycerol and 1-O-decyl-2-O-acetylglycerol, we showed that the ether bond was consistently associated with a loss of activity to varying extents. The results suggest that the ester bond in the 1-position is a major determinant of diacylglycerol-mediated protein kinase activation.

Animals↗
PubMed1987

Tumor promoter chloroform is a potent protein kinase C activator.

The major interaction site for tumor-promoting phorbol esters is the calcium-activated, phospholipid-dependent protein kinase (protein kinase C), a key-element in signal transduction. Binding of phorbol esters results in enzyme activation which mediates, at least in part, the action of these agents. We have investigated the effects of tumor promoter chloroform on protein kinase C activity. Like thrombin and 12-O-tetradecanoylphorbol-13-acetate (TPA), chloroform was able to activate protein kinase C in intact rabbit platelets. In addition, chloroform stimulated enzyme activity as well as TPA binding capacity in cell-free system. Scatchard analysis of the data has shown that chloroform increased the number of phorbol ester binding sites. Structurally related compounds, carbon tetrachloride and methylene chloride, activated the enzyme similarly.

Animals↗
PubMed1987

Mammary cancer antigen recognized by monoclonal antibody B72.3 in apocrine metaplasia of the human breast.

Monoclonal antibody B72.3 recognizing a pan-associated carcinoma antigen expressed also in metastatic human breast cancer cells has been tested using the avidin-biotin peroxidase method applied to paraffin-embedded sections in 50 samples of mammary tissue showing apocrine metaplasia and in 58 cases of other mild or severe focal epithelial proliferative changes of the breast, including mostly in situ lobular or ductal carcinomas collateral to clinical cancer removed after radical mastectomy. The antigen detected by this antibody was present in the apocrine cells of 48 cases (96%). In the majority of these cases the reactivity was localized on the luminal border of the apocrine cells and in the luminal secretion. But ten cases showed positive staining also in the cell cytoplasm either focal or diffuse. The normal structures and mild focal hyperplastic changes collateral to clinical cancer were, in the majority of the cases (43 of 55), negative, and, when positive, displayed positivity only at the luminal border. By contrast, the independent foci of in situ carcinoma (17 of 31 examined), the intraduct papillomas (seven cases of 14), and the intraductal component of breast carcinoma (seven cases of 17) were positive, displaying a cytoplasmic focal or diffuse staining. In conclusion, mammary apocrine metaplasia, a metaplastic change of the normal epithelium that has been associated with increased breast cancer risk, shares antigens in common with breast cancer cells and/or with cells showing severe atypia. The possible clinical significance of the site of antigenic expression (cytoplasm or luminal border) needs further investigation.

Antibodies, Monoclonal↗
PubMed1987

Benzene and toluene activate protein kinase C.

Protein kinase C plays a pivotal role in the transduction of signals controlling cell activation. A number of tumor promoters including phorbol esters activate the enzyme by substituting for diacylglycerol, the intracellular messenger of physiological ligands. In contrast, chloroform-induced protein kinase C activation does not appear to be mediated through the same mechanism. In vitro studies and experiments on intact platelets have revealed that benzene and toluene are also activators of protein kinase C. The data suggest that these drugs, which do not inhibit [3H]12-O-tetradecanoylphorbol-13-acetate binding to the enzyme, probably act in a chloroform-like manner.

Animals↗
PubMed1987

Phorbol esters as signal transducers and tumor promoters.

Extracellular ligands transfer information into the cell through various signaling pathways which operate in an integrated way. Oncogene proteins and tumor promoters cooperate to constitutively turn on the signaling pathways and lead to irreversible cell activation. The evidence that supports this concept is reviewed, with emphasis on the role of phorbol esters and protein kinase C in signal transduction and oncogenesis.

Animals↗
PubMed1985

Structural analogies between protein kinase C activators.

Phorbol esters and diacylglycerols activate protein kinase C but specific structural parameters appear to be required for the enzyme activation. We have analyzed the conformation of potent and not potent diacylglycerols and phorbol esters. The orientation of the CH20H group at C3 of 1,2 diolein is remarkably similar to that of the same group at C-20 of 4 beta phorbol didecanoate and crucial for potency in activating the enzyme. Our data suggest that the new conformational approach here described could be used to rationally design specific inhibitors preventing the effects of tumor promoters and to predict the structure of potential tumor promoters.

Acetylation↗
PubMed1985

Cholinergic and non-cholinergic forebrain projections to the interpeduncular nucleus.

A combined fluorescent retrograde tracing and acetylcholinesterase (AChE) histochemical technique was used for the study of some forebrain projections to the interpeduncular nucleus (IPN). After injections of a fluorescent tracer into the IPN, the distribution of AChE-containing and of fluorescent retrogradely labeled neurons was simultaneously studied in the habenular nuclei, medial septum and diagonal band of Broca. In all these regions, the majority of retrogradely labeled neurons also contained AChE: neurons located in the habenular nuclei stained lightly or moderately for the enzyme, while neurons located in the diagonal band and medial septum displayed intense AChE staining and were classified as putatively cholinergic perikarya. In all regions, a minority of labeled neurons did not stain for AChE, and were identified as non-cholinergic neurons projecting to the IPN. The present study shows the existence of a biochemical heterogeneity in the habenulo-interpeduncular and telencephalo-interpeduncular pathways, and indicates that the latter contains putatively cholinergic as well as non-cholinergic fibers.

Acetylcholinesterase↗
PubMed1985

Effects of tumor promoters on the expression of a tumor-related multigenic set in human cells.

We previously found that a minor subfraction of the human genomic DNA, corresponding to 2500-3000 nonrepetitive sequences of 3 kilobases each and designated as tumor-activated DNA (TaDNA) was transcriptionally active in Burkitt's lymphoma cells and almost inactive in normal lymphocytes growing in vitro following integration of the Epstein-Barr virus genome. Furthermore all the neoplastic cells in culture or primary neoplasms (leukemias, sarcomas, carcinomas) studied contained transcripts from most of the TaDNA sequences found in malignant lymphoblasts whereas normal cells growing in vitro contained only a few TaDNA transcripts. It is shown in the present study that treatments of the myeloid leukemia HL60 cells with various inducers of cell differentiation (dimethyl sulfoxide, retinoic acid, mezerein, 12-O-tetradecanoylphorbol-13-acetate, teleocidin) caused a dose-dependent reduction of the level of TaDNA transcripts, correlated with the diminution of c-myc transcripts. The 12-O-tetradecanoylphorbol-13-acetate treatment had this same effect on Burkitt's lymphoma cells (Raji or Namalwa) but the opposite effect on normal cells (Epstein-Barr virus-immortalized lymphocytes or fetal fibroblasts) where it enhanced the formation of Ta-DNA transcripts up to the levels found in untreated malignant cells. These data suggest two conclusions (a) TaDNA corresponds to a multigenic set which seems to be involved in modulation of the malignant phenotype and (b) depending on the origin of the cells, agents like 12-O-tetradecanoylphorbol-13-acetate may operate either as tumor promoters or as differentiation inducers through the control of TaDNA expression.

Actins↗
PubMed1984

Further characterization of tumor-promoter-mediated activation of protein kinase C.

Tumor promoting phorbol esters are able to activate Ca2+-sensitive, phospholipid-dependent protein kinase (protein kinase C) in a reconstituted system. Indol alkaloid teleocidin, a tumor promoter, has been found to be as potent as tumor promoters from the series of phorbol esters and mezerein in activating the mouse brain enzyme. Chemically unrelated tumor promoters such as tetrachlorodibenzo-p-dioxin, anthralin and phenobarbital are devoid of effect. Diacylglycerol 1,2 diolein strongly activated the enzyme whereas 1,3 diolein like 1,2 distearin were poor activators and 1,3 distearin was inactive. Although tumor-promoter-or diacylglycerol-mediated activation of protein kinase C was observed in the presence of 0.5mM EGTA, the reaction requires traces of Ca2+. Tumor promoters did not prevent inhibitory action of antipsychotic phenothiazines and local anesthetics but appear to increase IC50 of these drugs.

Animals↗
PubMed1984

Growth-rate-related and hydroxysterol-induced changes in membrane fluidity of cultured hepatoma cells: correlation with 3-hydroxy-3-methyl glutaryl CoA reductase activity.

3-hydroxy-3-methylglutaryl-coenzyme A reductase (EC 1.1.1.3.4.) activity and cell membrane fluidity measured by fluorescence polarization using 1,6 diphenyl, 1,3,5-hexatriene as fluorescent probe have been concomitantly examined in HTC hepatoma cells, both in relation to growth rate and in response to treatment with hydroxylated sterols. A high level of HMG-CoA reductase activity was observed in cells at log phase of growth which progressively decreased to reach a sustained low level at stationary phase. Similarly, membrane fluidity markedly decreased in relation to growth rate. Hydroxylated sterols such as 7 beta-hydroxycholesterol or 25-hydroxycholesterol strongly inhibited HMG-CoA reductase activity whereas a water-soluble derivative of 7 beta-hydroxycholesterol sodium 3,7-bishemisuccinate had no effect. Within the same range of concentrations 7 beta-hydroxycholesterol and 25-hydroxycholesterol strongly decreased membrane fluidity when the water-soluble derivative was ineffective. Thus, the present results provide evidence for a correlation between the two tested parameters and suggest a dependency of HMG-CoA reductase activity on cell membrane fluidity.

Animals↗
PubMed1984

Cutaneous metastasis of a radiation-associated thyroid medullary carcinoma.

The rare event of a single cutaneous metastasis occurred in a 37-year-old white man two years after a subtotal thyroidectomy for sporadic medullary carcinoma is reported. The endocrine nature of the tumor is revealed by the ultrastructural study, and the type of secretion by the biochemical assay of calcitonin (CT) in the serum and in the tumor tissue extract and by the PAP (peroxidase-anti-peroxidase) technique with anti-CT antibodies on paraffin sections. The cytological and histological findings are described. Some peculiar ultrastructural aspects are presented in details. The fact that the patient received a course of Rx therapy (3,000 rads) on the neck for cervical syringomyelia at the age of 20 years is stressed.

Adult↗
PubMed1984

Thermal tolerance by embryos and larvae of the surf clam Spisula solidissima.

Thermal tolerance was tested in three developmental stages of the surf clam Spisula solidissima: cleavage stages, trochophore , and straight hinge larvae. Experiments were designed to simulate larval entrainment in the cooling system of a power plant. Eleven different temperatures were investigated from 20.2 to 43 degrees C. Exposure times varied from 1 min to 6 hr and following each of the (88) different exposures, larvae were returned to the incubation temperature for a further period of development which facilitated differentiation of dead and live animals. Pooled mortality data from triplicate experiments (for each larval state) were subjected to a stepwise regression analysis (percentage mortality on temperature and time). Equations were derived from the regression program comprising first-, second-, and third-order terms for temperature and time, and these were used to generate response surfaces relating mortality to temperature-time exposure. As with earlier work on Mulinia , the term (temperature) provides the best single predictor of larval mortality. Spisula is the only fully marine bivalve species so far subjected to this type of thermal tolerance test and is clearly the most temperature sensitive of all species investigated to date.

Animals↗
PubMed1984

Morphologic assessment of reactivity to monoclonal antibodies generated against breast cancer cells in mammary tissues removed for clinical dysplasia or cancer.

Monoclonal antibodies B 6.2, B 72.3 and B 1.1, reactive against tumor-associated antigens of human breast cancer, have been tested on histologic sections of paraffin blocks from 49 cases of breast cancer and from 13 cases of mammary dysplasia. The results have shown that: (a) the three monoclonal antibodies recognize different antigenic determinants of the cancer cells; (b) monoclonal antibodies B 6.2 and B 1.1 recognize antigens which are frequently expressed in cancer cells; (c) the antigen recognized by monoclonal antibody B 72.3, at the concentrations used, is less frequently expressed in tumors, while it seems to be selective for the apical cell border and luminal secretions of apocrine metaplastic lobules or cysts occurring in mammary dysplasia; (d) intracytoplasmic expression of the antigens is observed only in cancer cells, with rare exceptions; (e) tumor cell populations are heterogeneous in respect to the expression of the antigens detected by the monoclonal antibodies tested; (f) the antigens detected are unrelated to the tumor histotype; (g) the frequency of axillary lymph nodal metastases is not influenced by the presence or intensity of antigenic expression in the primary tumors; but (h) in metastatic tumor cell populations the antigenic expression is higher than in primary tumors. This fact suggests some relationship between tumor metastatization and the expression of antigens recognized by the monoclonal antibodies tested.

Adult↗
PubMed1984

Protein phosphorylation and mechanism of action of tumor-promoting phorbol esters.

The phorbol ester TPA may be intercalated into the membrane phospholipid bilayer and selectively binds to every molecule of C-kinase in exhibiting its full enzymatic activity. Available evidence suggests that C-kinase is the receptive protein of this tumor promoter, and the results presented seem to provide clues for clarifying the mechanism of controlling cell growth and differentiation.

Animals↗
PubMed1983

Synergistic functions of phorbol ester and calcium in serotonin release from human platelets.

In human platelets, thrombin activates Ca2+-activated, phospholipid-dependent protein kinase (protein kinase C) and mobilizes Ca2+ concomitantly, whereas 12-O-tetradecanoylphorbol-13-acetate (TPA) may be intercalated into membranes and directly activates protein kinase C without mobilization of Ca2+ in sufficient quantities. A series of experiments with TPA and Ca2+-ionophore (A23187) indicates that activation of protein kinase C is a prerequisite requirement for release of serotonin, and that this enzyme activation and Ca2+ mobilization act synergistically to elicit a full cellular response. Both cyclic AMP and cyclic GMP inhibit activation of protein kinase C by prohibiting the signal-dependent breakdown of inositol phospholipid to produce diacyl-glycerol, but none of these cyclic nucleotides prevents the TPA-induced activation of this enzyme.

Blood Platelets↗