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Biomedical subjects

F Mannello

Publications and source records attributed to F Mannello.

At least 37 records · Page 2Linked to original sources
PubMed1999

Immunoreactivity, ultrastructural localization, and transcript expression of prostate-specific antigen in human neuroblastoma cell lines.

Prostate-specific antigen (PSA) is considered a highly specific biochemical marker of the human prostate gland, and it currently is used for prostate cancer diagnosis and monitoring. Recently, PSA production and secretion were found in nondiseased and diseased cells, tissues, and fluids from women. In this study, we characterized the presence of PSA in two human neuroblastoma cell lines with biochemical, ultrastructural, and molecular approaches. Using reverse transcription-PCR, we identified PSA mRNA, and Western blotting revealed a substantial amount of complexed form of PSA protein, which is localized mainly in free ribosomes. Although the role of PSA in human neuroblastoma cell lines is still unknown, our study supports the hypothesis that this serine protease may be involved in controlling the growth of human brain tumor cells, adding more support to the notion that PSA is a widespread kallikrein-like protease with biological functions much more complex than recently thought.

Blotting, Western↗
PubMed1998

Ultrastructural characterization and biochemical profile of human gross cystic breast disease.

Human gross cystic breast disease is a benign condition affecting about 7-10% of adult women occurring with the highest incidence in the premenopausal decade. Although breast cysts do not represent a preneoplastic condition per se, several studies indicate an increased breast cancer risk in women affected by this pathology. In this report we study 115 breast cystic fluid samples obtained by needle-aspiration from women with gross cystic breast disease. The samples were analysed biochemically and the cells contained therein were observed at the electron microscope. According to their biochemical profiles, the cysts were subdivided into three types: Type I, showing a Na/K ratio < 0.5 and a typical protein content; Type II, showing a Na/K ratio >10 and a protein content quite similar to plasma; Type III, showing a Na/K ratio between 1 and 7 and an intermediate protein content. The electron microscopic examination demonstrated that Type I cystic fluid cells exhibit morphological features typical of actively synthesising and secreting cells, while the characteristics of Type II cells indicate a low metabolic activity. Type III cells have characteristics typical of both Type I and Type II cells, thereby confirming the intermediate nature of this cyst type. We hypothesise that these cyst types could represent different developmental stages of a structural evolution pathway, during which the biosynthetically active 'apocrine stage' would be the key step to cell neoplastic transformation.

Adult↗
PubMed1997

Prostate-specific antigen expression in a case of intracystic carcinoma of the breast: characterization of immunoreactive protein and literature surveys.

A case is presented of female breast intracystic carcinoma with prostate-specific antigen (PSA) expressed in high amounts in aspirated cystic fluid (55 micrograms/L). Tumor extract analysis revealed the presence of both estrogen and progesterone receptors (0.38 and 1.87 nmol/L, respectively) and high quantities of PSA too (19.52 micrograms/L). Chromatographic analysis of cystic fluid revealed two peaks of PSA, at the expected positions for free and bound serine protease. A major proportion of 33-kDa free from was also confirmed by Western blotting analysis. Free PSA was heat-stable at 56 degrees C and displayed no change after freezing-thawing. These findings are discussed in the context of a detailed literature survey. Our data support the contention that PSA immunoreactivity in intracystic fluid of breast carcinoma is partly the result of secretory activity by the neoplastic cells and that the steroid receptors can also modulate its expression.

Aged↗
PubMed1996

Lipids status in human breast cyst fluids.

Benign mammary gross cystic disease is the most common breast lesion; women with apocrine changes of epithelium lining the cysts are at higher risk for developing breast cancer than the normal population. Total cholesterol, high- and low-density lipoproteins fractions, triglycerides and phospholipids, lipase activity and total lipid concentrations were measured in cyst fluids and sera from 89 women affected by gross cystic breast disease. Total cholesterol and high-density lipoprotein content were significantly (P < 0.001) greater in pooled cyst fluids than normal sera. Moreover, data analyses show a significant increase in the mean values of total lipids and lipase activity in metabolically active apocrine cysts, when compared to the flattened cysts (P < 0.001). The lipids feature of apocrine cysts could represent an altered expression of biosynthetic activity of the surrounding apocrine cell surface glycolipid and steroidogenic metabolism and may provide further knowledge about the functional stage changes of gross breast cysts.

Adult↗
PubMed1996

Quantification of prostate-specific antigen immunoreactivity in human breast cyst fluids.

The frequency of gross cystic breast disease in premenopausal women and its possible association with increased breast cancer risk emphasises the importance of investigations relating to breast cyst fluid composition. In order to contribute to a better analysis of this medium, we have measured the presence of prostate-specific antigen immuno-reactivity in sixty-four human breast cyst fluids. Data analyses show that 35% of samples presented a level of this antigen < 0.05 micrograms/L, whereas 42 out of 64 cysts show a significant increase in the mean value of metabolically active apocrine cysts when compared to flattened cysts (p < 0.01). We report the first evidence that breast epithelium of gross cysts produces, secretes, and accumulates large amounts of prostate-specific antigen, a glycoprotein produced by prostatic tissue but recently detected in breast tumours, normal tissues, and during pregnancy. The production and intracystic accumulation of this serine protease in biosynthetically active apocrine type cyst can play a feasible role in the natural history gross cystic breast disease as well as in the mechanism of cyst formation, enlargement, and transformation.

Adult↗
PubMed1996

Promotion of tumour metastases and induction of angiogenesis by native HIV-1 Tat protein from BK virus/tat transgenic mice.

OBJECTIVE: To characterize the T53 cell line and its clones derived from an adenocarcinoma of BK virus (BKV)/tat transgenic mice and to establish the role of native Tat in tumorigenicity, induction of metastases and angiogenesis. DESIGN AND METHODS: Tat was quantified by flow cytometry and chloramphenicol acetyltransferase (CAT) assays. Tumorigenicity and metastatic ability of cell lines were assayed in nude mice. Production of proteases was evaluated by a plasmin chromogenic assay and gelatinase zymography. The angiogenic effect was studied in vivo with conditioned medium from tumour cell lines. RESULTS: Tat protein was detected in tumour cell lines in amounts from 600-7000 molecules/cell. Conditioned medium from tumour cell lines was able to transactivate an LTR-CAT in HL3T1 cells, indicating release of extracellular Tat. Tumour cell lines, inoculated into nude mice induced angiogenic tumours with remarkable recruitment of host endothelial cells. Metastases were detected in lymph nodes, lungs, kidneys, and heart. Cell lines produced relevant amounts of proteases. Conditioned medium implanted in mice with matrigel induced an angiogenic response, enhanced by addition of heparin. Preincubation with an anti-Tat antibody abolished the angiogenic effect. CONCLUSIONS: Tat from cells from BKV/tat transgenic mice promotes tumorigenesis and formation of metastases and induces angiogenic activity. Angiogenesis occurs at physiological concentrations of Tat lower than 20 ng/ml. The effects of Tat on induction of metastases and angiogenesis appear to be mediated by activation of proteases.

Animals↗
PubMed1995

beta-Endorphin expression in gross cystic breast disease.

Opioid peptides have a variety of pathophysiologic actions, playing a novel important role in human breast cancer. The expression of beta-endorphin was studied in 84 human breast cyst fluids from gross cystic breast disease-affected patients. The concentration of beta-endorphin in pooled breast cyst fluids was over four-fold higher than in respective plasma with a significant increase in the mean value of the 'metabolically active' apocrine cysts when compared with flattened cysts (P < 0.001). The higher levels of Type I cyst suggest de novo mammary synthesis of endogenous opioid peptides and could represent an altered expression of biosynthetic activity of apocrine breast cells, providing a possible explanation on functional changes of gross cysts, on the mechanism of their formation and a perspective relationship to breast cancer risk.

Adult↗
PubMed1994

Alpha 1-antichymotrypsin complexes in human breast cyst fluids.

Alpha 1-antichymotrypsin, a serum protease inhibitor, was found in 72 breast cyst fluids aspirated from women affected by gross cystic breast disease. When fractionated by gel chromatography, the presence of protein complexes or aggregates was demonstrated. A different distribution of the alpha 1-antichymotrypsin appeared to be related to the ionic composition of the breast cyst fluid; when compared with metabolically active apocrine cysts, a statistically significant increase of alpha 1 protease inhibitor values in flattened epithelial cysts was revealed (P < 0.001). Two-dimensional immunoelectrophoresis showed in apocrine cysts (Na/K ratio < 3) a characteristic double peak of alpha 1-antichymotrypsin immunoprecipitin curve. The relationship between this alpha 1 protease inhibitor and electrolyte profiles may provide further knowledge about the imbalance between proteases and their inhibitors on functional changes of gross cysts and might be useful in studies on their mechanism of formation and relationship to subsequent breast cancer.

Exudates and Transudates↗