Immunoreactive prostate-specific antigen in pleural effusions.
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Aged↗
Biomedical subjects
G Gazzanelli
Publications and source records attributed to G Gazzanelli.
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↗
Prostate-specific antigen in ascitic fluid.
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Adolescent↗
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↗
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↗
Non-invasive early prenatal molecular diagnosis using retrieved transcervical trophoblast cells.
Fetal DNA was recovered from 17 of 39 (44%) transcervical cell (TCC) samples obtained between 7 and 9 weeks of gestation by endocervical canal flushing. Trophoblast retrieval was adequate for polymerase chain reaction (PCR) amplification of Y chromosome-specific DNA sequences and detection of paternal-specific microsatellite alleles. The fetal sex predicted by PCR in TCCs was confirmed in all cases by karyotype analysis of chorionic villi at 10 weeks of gestation. The absence of the disease-associated paternal alleles in TCC samples from two pregnancies at risk for spinal muscular atrophy and myotonic dystrophy predicted unaffected fetuses in agreement with subsequent results on chorionic villi and newborns' leukocytes. A trisomy 21 fetus was diagnosed in TCCs using fluorescent in situ hybridization (FISH) and semi-quantitative PCR analysis of superoxide dismutase-1 (SOD 1). Present experience indicates that TCC sampling is a promising technique for early prenatal monitoring of Mendelian disorders and chromosome aneuploidy.
Adult↗
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↗
Immunoreactivity of prostate-specific antigen in plasma and saliva of healthy women.
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Adult↗
Breast cancer diagnosis by lactate dehydrogenase isozymes in nipple discharge.
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Breast Diseases↗
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↗
2',3'-Dideoxycytidine induced drug resistance in human cells.
2',3'-Dideoxycytidine (ddC) is a nucleoside analogue that inhibits HIV-1 replication in vitro and is currently used in AIDS therapy. This compound exerts a delayed cytotoxicity due to inhibition of mitochondrial DNA (mDNA) synthesis. We have found that long term exposure of U937 human monoblastoid cells to ddC allowed the selection of a drug-resistant cell line (U937-R) with 66% mDNA, normal ddC transport and altered deoxycytidine kinase kinetic properties. In this paper we show that U937-R cells contain an increased number of mitochondria per cell and a reduced copy number of mDNA/mitochondria. Furthermore, the intracellular concentrations of deoxycytidine 5'-triphosphate (dCTP) and 2',3'-dideoxycytidine 5'-triphosphate (ddCTP) are also reduced although with a higher dCTP/ddCTP ratio in U937-R compared to the parental cells. This mechanism of drug resistance, with drug-resistance based on viral mutations, can provide an explanation for drug failure in antiviral therapy.
Cell Line↗
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↗
Commissural connections of the cat periaqueductal gray matter studied with anterograde and retrograde tract-tracing techniques.
The commissural connections of the periaqueductal gray matter were investigated by light and electron microscopy by using the anterograde tracer Phaseolus vulgaris leucoagglutinin and the retrograde tracer horseradish peroxidase. In the first group of seven animals (1-7), single injections of Phaseolus vulgaris leucoagglutinin were performed iontophoretically (4.5 microA for 30 min) into various subdivisions of the periaqueductal gray matter. On light microscopic examination, injection sites were characterized by several immunolabeled neurons of different sizes and morphology, with the cytoplasm, nucleus and neuronal processes intensely stained. Many labeled fibers turned from injection sites toward all contralateral periaqueductal gray matter subdivisions, but anterograde labeling was densest in the regions homotopic to those injected. Commissural fibers bore along their course many en passant boutons of different sizes and morphology, and gave off spine-like processes, at the end of which one terminal bouton was observed. Labeled fibers branched into numerous collaterals which ended in a terminal array of 10-20 en passant and en grappe boutons. At the electron microscopic level, commissural axons were observed in close proximity to the cytoplasmic membranes of cells. Axon terminals formed symmetric or asymmetric synapses mainly on dendritic shafts of neurons and rarely on vesicle-containing profiles. Horseradish peroxidase experiments were carried out in four cats (1-4). The tracer was injected iontophoretically into different regions of the periaqueductal gray matter of three cats (1-3). Retrogradely labeled neurons giving rise to commissural connections had a morphology similar to that of polygonal, triangular and fusiform cells described in previous Golgi studies. The perikaryal cross-sectional area of commissural neurons was smaller than that of neurons projecting outside the periaqueductal gray matter (mean value of commissural neurons 149.77 microns 2 vs 261.19 microns 2 for projecting neurons), which were retrogradely labeled by pressure-injecting horseradish peroxidase into several targets of periaqueductal gray matter (4). Moreover, since the distribution of sizes of the two populations of the periaqueductal gray matter overlapped in the range of 90-300 microns 2, a considerable number of projecting neurons were as small as commissural neurons. The present results suggest that commissural fibers could reciprocally connect zones of the periaqueductal gray matter with similar functions, and originate from small and medium-sized neurons, some of which are also projecting neurons.
Animals↗
Hydrogen peroxide cytotoxicity under conditions of normal or reduced catalase activity in H2O2-sensitive and -resistant Chinese hamster ovary (CHO) cell variants.
H2O2-sensitive and -resistant sublines of Chinese Hamster Ovary (CHO) cells were tested for their sensitivity to the growth inhibitory effect elicited by increasing concentrations of the oxidant under conditions of normal or reduced catalase activity. Experimental results have demonstrated that, under conditions of reduced catalase activity, the cytotoxic action of H2O2 was differentially regulated in resistant and sensitive cells. Indeed, the parental cell line and cells resistant to low concentrations of H2O2 (V 250 cells) depended on catalase to a lower extent than did highly resistant cells (V 850 cells). It is interesting to note that V 250 cells had more catalase, on a per million cell basis, than V 850 cells. We conclude that acquired resistance to oxidative stress is not entirely dependent on catalase and that the contribution of catalase depends on the degree of resistance to the oxidant.
Animals↗
Development and characterization of hydrogen peroxide-resistant Chinese hamster ovary cell variants--I. Relationship between catalase activity and the induction/stability of the oxidant-resistant phenotype.
Hydrogen peroxide (H2O2)-resistant sublines of Chinese hamster ovary (CHO) cells were isolated by in vitro exposure to the oxidant (treatment for 1 hr followed by 3 days of growth in peroxide-free medium). Stepwise increase in low level H2O2 concentrations produced variants which were progressively more resistant to the growth inhibitory effect elicited by the oxidant. Removal from H2O2 decreased resistance and the curve describing this process was biphasic in nature. In addition, the rate of loss of the H2O2-resistant phenotype was more rapid for the toxicity elicited by low concentrations of hydrogen peroxide, compared to that produced by high concentrations. Changes in total cell proteins were found to parallel the variations in sensitivity to the oxidant, since the protein content constantly increased during the adaptation process and decreases upon removal from H2O2. Catalase activity did not show large variations in resistant sublines with respect to the parental cell line, and these changes were at least partially related to differences in cell size/amount of total cell proteins of the sublines. In addition, the minor changes observed for catalase activity did not correlate with the degree of resistance to growth inhibition elicited by the oxidant. It may therefore be suggested that the H2O2-resistant phenotype of mammalian cells, initially adapted to low--then gradually increased--concentrations of the oxidant, is the result of a complex phenomenon which only partially involves over-expression of catalase.
Animals↗
Intracellular distribution of hexokinase in rabbit brain.
Hexokinase in mammalian brain is particulate and usually considered to be bound to the outer mitochondrial membrane. Investigation of rabbit brain mitochondria prepared either by differential centrifugation and discontinuous density gradient centrifugation has provided evidence that this particulate fraction also contains endoplasmic vesicles and synaptosomes. Solubilization of the bound hexokinase by different combinations of detergents and metabolites has proved the existence of different hexokinase binding sites. Electron microscopic examination of hexokinase location by immuno-gold labelling techniques confirmed that hexokinase is indeed predominantly bound to mitochondria but that a significant proportion is also bound to non-mitochondrial membranes. Attempts to quantify this distribution were unsuccessful since different figures were obtained using anti-hexokinase IgG affinity purified on immobilized native or denatured hexokinase. Binding studies of the purified rabbit brain mitochondrial hexokinase to rabbit liver mitochondria and microsomes confirmed that in addition to a binding site on mitochondria there is another binding site on microsomes. The N-terminal sequence of hexokinase has been shown to be important for mitochondria binding and also for microsome binding. These results suggest that the intracellular localization of hexokinase in rabbit brain is not exclusively mitochondrial and that the metabolic role of this enzyme should be reconsidered by including a binding site on the endoplasmic reticulum.
Animals↗
Establishment and characterization of two human small cell lung carcinoma cell lines.
Two human cellular lines of cancerous small lung cells obtained from biopsies specimens from two patients were characterized in vitro according to cellular morphology, growth modality, caryotype and antigenic profile. In vivo studies were carried out by inoculating the cells into nude mice of both sexes and various ages by different routes in order to study tumorigenicity and metastatic capacity and to identify a biological marker of the malignancy. Results obtained to date suggest that the two cellular lines have different biological properties similar to the classic and the variant form found in literature. A biological marker of the malignancy seems to be the antigenic profile of the cells.
Age Factors↗
Lipid-associated sialic acid levels 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 female population. Sialic acid has drawn considerable interest because of carbohydrate aberrations in malignant cells. The current investigation determined the concentrations of lipid-associated sialic acid (LASA) in 62 breast cyst fluids and sera. Data analyses show a significant increase in the mean values of LASA in metabolically active apocrine cysts when compared to the cysts with Na+/K+ > 3 (flattened cysts) (p < 0.001). The greater LASA levels in cyst fluids with lower intracystic Na+/K+ ratios could represent an altered expression of biosynthetic activity of the surrounding apocrine cell surface sialoglycolipid metabolism, providing a possible explanation of why women with apocrine cysts may be at greater cancer risk and being useful in further studies on functional stage changes in the cysts and their relationship to breast cancer.
Adult↗