Objective An increased percentage of Vδ1+/γ/δT cells has been detected both in the peripheral blood and bronchoalveolar lavage fluid of patients with systemic sclerosis (SSc). This study evaluated the subset distribution, activation... more
Objective An increased percentage of Vδ1+/γ/δT cells has been detected both in the peripheral blood and bronchoalveolar lavage fluid of patients with systemic sclerosis (SSc). This study evaluated the subset distribution, activation status, and expression of cellular ...
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The incidence of anti-myocardial autoantibodies was investigated in 62 patients suffering from congenital or acquired cardiopathy prior to and 1, 10 and 20 days after cardiac surgery. Anti-myocardial autoantibodies were present in about... more
The incidence of anti-myocardial autoantibodies was investigated in 62 patients suffering from congenital or acquired cardiopathy prior to and 1, 10 and 20 days after cardiac surgery. Anti-myocardial autoantibodies were present in about 23.7 per cent of patients even before surgery, practically doubled one day after intervention, reached their peak (60-70 per cent) at 10 days from surgery and began to diminish at 20 days. At all times and incidence was higher in patients with acquired cardiopathy than in those with congenital heart diseases. In the large majority of cases, the autoantibodies were of the IgG class, of the striational type and in low titers. The relevance of anti-myocardial autoantibodies in the pathogenesis of cardiac lesions and the possible causes of the reversibility of the autoimmune phenomenon are discussed.
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We have established an ovarian cancer cell line (CABA I) from ascitic fluid obtained from a patient with papillary adenocarcinoma of the ovary prior to drug treatment. The epithelial origin of the cell line was confirmed by morphology and... more
We have established an ovarian cancer cell line (CABA I) from ascitic fluid obtained from a patient with papillary adenocarcinoma of the ovary prior to drug treatment. The epithelial origin of the cell line was confirmed by morphology and by immunofluorescence analysis using anticytokeratin antibodies. Ultrastructural analysis revealed a very irregular membrane surface and a clear cytoplasm rich in electron-lucent vesicles. CABA I cells grow rapidly in culture (doubling time 18 h) in an anchorage-independent manner. Exogenously added beta-estradiol and epidermal growth factor (EGF) treatments did not influence cell growth rate. FACS analysis to determine the phenotypic profile of tumor-associated antigen, membrane receptor, and adhesion molecule expression indicated that the cell line was positive for different members of the c-erbB family, for alpha 6 and beta 1 integrin receptors, and intensively positive for HLA class I antigens and the folate receptor. Molecular characterization revealed no mutations for c-myc and c-k-ras genes, but did detect an exon 5 mutation in the p53 gene. CABA I cells grew poorly as heterotransplants in nude mice, and tumors showed long latency periods. Because early (15-20) and late (55-60) passage cells maintain the same growth and phenotypic characteristics, the CABA I cell line might provide a good in vitro model system to investigate the cellular and molecular events involved in ovarian carcinogenesis.
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Research Interests: Metabolism, Cancer, Nuclear Magnetic Resonance, Biology, Medicine, and 15 moreCell line, Humans, Female, Enzyme, Disease Progression, Phospholipase D, Choline, Epithelial Cell Biology, Phosphatidylcholine, Confidence Limit, Tumor Progression, Phospholipase C, Choline Kinase, phosphorylcholine, and Ovarian Neoplasms
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IntroductionThe use of Performance and Image-Enhancing Drugs (PIEDs) is on the increase and appears to be associated with several psychopathological disorders, whose prevalence in unclear.Objectives/AimsWe aimed to evaluate the... more
IntroductionThe use of Performance and Image-Enhancing Drugs (PIEDs) is on the increase and appears to be associated with several psychopathological disorders, whose prevalence in unclear.Objectives/AimsWe aimed to evaluate the differences–if any–in the prevalence of body image disorders (BIDs) and eating disorders (EDs) in PIEDs users athletes vs. PIEDs nonusers ones.MethodsWe enrolled 84 consecutive professional and amateur athletes (35.8% females; age range = 18–50), training in several sports centers in Italy. They underwent structured interviews (SCID I/SCID II) and completed the Body Image Concern Inventory (BICI) and the Sick, Control, One, Fat, Food Eating Disorder Screening Test (SCOFF). Mann-Whitney U test and Fisher's exact test were used for comparisons.ResultsOf the 84 athletes, 18 (21.4%) used PIEDs. The most common PIEDs were anabolic androgenic steroids, amphetamine-like substances, cathinones, ephedrine, and caffeine derivatives (e.g. guarana). The two groups did not differ in socio-demographic characteristics, but differed in anamnestic and psychopathological ones, with PIEDs users athletes being characterized by significantly (P-values < 0.05) higher physical activity levels, consuming more coffee, cigarettes, and psychotropic medications (e.g. benzodiazepines) per day, presenting more SCID diagnoses of psychiatric disorders, especially Substance Use Disorders, Eating Disorders, Body Dysmorphic Disorder (BDD), and General Anxiety Disorders, showing higher BICI scores, which indicate a higher risk of BDD, and higher SCOFF scores, which suggest a higher risk of BIDs and EDs.ConclusionsIn PIEDs users athletes body image and eating disorders, and more in general psychopathological disorders, are more common than in PIEDs nonusers athletes.Disclosure of interestThe authors have not supplied their declaration of competing interest.
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CT-PET scan showing clinical response of Pt#3
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CT-PET scan showing clinical response of Pt#2.
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Purpose:This study was aimed at evaluating the feasibility, safety, immunologic and clinical responses in patients with follicular lymphoma treated with monocyte-derived dendritic cells generated in the presence of IFNα and GM-CSF... more
Purpose:This study was aimed at evaluating the feasibility, safety, immunologic and clinical responses in patients with follicular lymphoma treated with monocyte-derived dendritic cells generated in the presence of IFNα and GM-CSF (IFN-DC) in combination with low doses of rituximab.Patients and Methods:Firstly, we analyzed in vitro and in vivo the immunologic properties of IFN-DC against follicular lymphoma. Thus, we performed a phase I trial in 8 patients with refractory and relapsed follicular lymphoma based on sequential intranodal injections of low-dose of rituximab and unloaded IFN-DC and report the safety, clinical, and immunologic results of the enrolled patients.Results:Preclinical studies indicated that IFN-DC can synergize with rituximab leading to increased cytotoxicity and T-cell tumor infiltration. The clinical evaluation showed that the combined treatment was totally safe. The overall response rate was 50%, PET-negative complete response rate 37%, and remission is stil...
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The combination of freeze-fracture and immunocytochemical techniques permits to analyze the distribution of membrane components on both plasma and intracellular membranes (1–3). In particular the fracture-label approach, where cells are... more
The combination of freeze-fracture and immunocytochemical techniques permits to analyze the distribution of membrane components on both plasma and intracellular membranes (1–3). In particular the fracture-label approach, where cells are directly immunolabeled after freeze-fracture allows the exposition to the immunolabeling of the freeze-fractured inner nuclear membrane. Moreover, since during freeze-fracture, transmembrane proteins partition with the inner (protoplasmic) or outer (exoplasmic) leaflets of the freeze-fractured membrane, the fracture-label approach can also assess this partition. Therefore, we decided here to apply fracture-label technique to EBV producer lymphoblastoid cells in order to investigate the partition on fracture, distribution and extent of glycosylation of EBV envelope glycoproteins during the budding process.
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SUMMARYThis study provides evidence that cardiolipin (CL) molecules are expressed on the surface of apoptotic cells and are recognized by antiphospholipid antibodies, purified from patients with the antiphospholipid antibody syndrome... more
SUMMARYThis study provides evidence that cardiolipin (CL) molecules are expressed on the surface of apoptotic cells and are recognized by antiphospholipid antibodies, purified from patients with the antiphospholipid antibody syndrome (APS). CL expression on cell surface was demonstrated by high performance thin layer chromatography analysis of phospholipids from plasma membrane purified fractions and by the positive staining with the CL-specific dye nonyl-acridine orange. This finding was complemented with the observation that aCL IgG purified from patients with APS bind to the surface of apoptotic cells. This staining shows a clustered distribution mostly localized on surface blebs. Interestingly, CL exposure on the cell surface preceded the DNA fragmentation, as shown by cytofluorimetric analysis. These findings demonstrate that exposure of CL molecules on the cell plasma membrane is an early event of the apoptotic cellular program that may represent an in vivo trigger for the gen...
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Supplementary Table I: IFN-DC release criteria and median values registered among patient-derived IFN-DC (min-max range); Supplementary Table 2: Patients MHC-I and MHC-II alleles and IGHV epitope sequences
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Despite advances in pharmacotherapy of lipid disorders, many dyslipidemic patients do not attain sufficient lipid lowering to mitigate risk of atherosclerotic cardiovascular disease. Several classes of novel lipid-lowering agents are... more
Despite advances in pharmacotherapy of lipid disorders, many dyslipidemic patients do not attain sufficient lipid lowering to mitigate risk of atherosclerotic cardiovascular disease. Several classes of novel lipid-lowering agents are being evaluated to reduce atherosclerotic cardiovascular disease risk. Lipoprotein apheresis (LA) is effective in acutely lowering the plasma concentrations of atherogenic lipoproteins including low-density lipoprotein cholesterol and lipoprotein(a), and novel lipid-lowering drugs may dampen the lipid rebound effect of LA, with the possibility that LA frequency may be decreased, in some cases even be discontinued. This document builds on current American Society for Apheresis guidelines and, for the first time, makes recommendations from summarized data of the emerging lipid-lowering drug classes (inhibitors of proprotein convertase subtilisin/kexin type 9 or microsomal triglyceride transfer protein, high-density lipoprotein mimetic), including the avai...
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The surface ultrastructure of lymphocytes during capping of two transmembrane proteins is shown. As seen by fracture-flip the plasma membranes of human lymphocytes are covered by a high density of surface particles. Incubation in 30%... more
The surface ultrastructure of lymphocytes during capping of two transmembrane proteins is shown. As seen by fracture-flip the plasma membranes of human lymphocytes are covered by a high density of surface particles. Incubation in 30% glycerol leads to aggregation of these surface particles. Immunogold labelling shows that the transmembrane proteins bearing HLA class I and CD4 antigens are confined to the particle aggregates. These results indicate that surface particles revealed by fracture-flip represent surface protrusions of integral membrane proteins seen as intramembrane particles in freeze-fractured lymphocytes. During capping HLA or CD4 antigens aggregate into progressively larger patches and, finally, into single caps. As revealed by fracture-flip the patches/caps are seen as clearly differentiated raised platforms that are clearly and sharply demarcated relative to contiguous areas of the surface. In non-patched (non-capped) regions, the pattern of distribution and apparent...
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Paclitaxel (PTX) is a potent anti-neoplastic agent that is highly effective in treating ovarian cancer. Nevertheless, the emergence of PTX resistance has limited the control of this disease. To gain insight into the molecular alterations... more
Paclitaxel (PTX) is a potent anti-neoplastic agent that is highly effective in treating ovarian cancer. Nevertheless, the emergence of PTX resistance has limited the control of this disease. To gain insight into the molecular alterations accompanying drug resistance in ovarian cancer, we generated a new stable PTX-resistant ovarian carcinoma cell line. CABA I cells, which display an intrinsic PTX resistance (IC50 = 800 ng/ml), were subjected to continuous exposure to PTX. From the residual surviving cells, the highly PTX-resistant line CABA-PTX (IC50 = 256000 ng/ml) was generated and stably maintained in vitro. Analysis of beta-tubulin expression indicated that only the HM40 and Hbeta9 isotypes were expressed in both parental and resistant cells. No specific point mutations in the HM40 were detected in either cell line, but expression levels of this isotype were significantly reduced (40%) in CABA-PTX cells. Hbeta9 levels were unchanged. In those cells, PTX resistance was associated...
Research Interests: Flow Cytometry, Biology, Cancer Research, Ovarian Cancer, Medicine, and 15 moreCell Culture, Cell Division, Humans, Multidrug Resistance, Female, Paclitaxel, Ribosomal proteins, Phenotype, Time Factors, Methotrexate, Tubulin, Paclitaxel Resistance, Mitochondrial Proteins, fluorouracil, and Ovarian Neoplasms
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Purpose: We have developed a novel DC population differentiated from human monocytes in the presence of GM-CSF and IFN-alpha (IFN-DC), which is highly efficient in internalizing tumour cell antigens and in the cross-priming of CD8+T... more
Purpose: We have developed a novel DC population differentiated from human monocytes in the presence of GM-CSF and IFN-alpha (IFN-DC), which is highly efficient in internalizing tumour cell antigens and in the cross-priming of CD8+T cells, promoting efficient anti-lymphoma response. Preclinical results prompted us to start a clinical trial of IFN-DC combined with low-dose Rituximab(RTX) in patients with Follicular lymphoma (FL), which is an indolent, immune-responsive, but still incurable cancer. Patients and Methods: In October 2014 we started a phase I clinical trial (EudraCT: 2013-003158-25) testing safety, immunogenicity and systemic clinical responses of low-dose RTX combined with autologous IFN-DC, administered by intra-nodal injections. Patients features: age 18-75y, indolent FL, relapsed or refractory (R/R), stage III-IV low tumour burden, with at least 1 superficial pathologic lymph node. One leukapheresis was scheduled at study entry for harvesting mononuclear cells. Eight...
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This study was undertaken to analyze both the GM3 expression on peripheral blood lymphocytes of HIV-infected patients and the relationship between ganglioside content and anti-GM3 reactivity. GM3 expression was determined as a percentage... more
This study was undertaken to analyze both the GM3 expression on peripheral blood lymphocytes of HIV-infected patients and the relationship between ganglioside content and anti-GM3 reactivity. GM3 expression was determined as a percentage of lipid-bound sialic acid and by cytofluorimetric analysis in 25 AIDS patients, 20 anti-HIV+ asymptomatic subjects, 25 patients with different viral disease, and 25 healthy donors. GM3 distribution was analyzed by immunofluorescence and immunoelectron microscopy. A follow-up study to detect anti-lymphocytic GM3 antibodies was performed in progressive and nonprogressive anti-HIV+ subjects. Lymphocytes from HIV-infected patients showed a significant increase of plasma membrane GM3 content; no difference was found between CD4+ and CD8+ cells. Immunofluorescence and immunoelectron microscopic analysis showed that GM3 was distributed in large clusters over the cell plasma membrane. The follow-up study revealed that the occurrence of anti-lymphocytic GM3 antibodies was significantly higher in patients with progressive disease, compared with asymptomatic non-progressive subjects. These findings revealed that (1) the increased GM3 content in HIV-infected patients is detected at the plasma membrane level, (2) GM3 overexpression is able to induce an increased reactivity with anti-GM3 antibodies, and (3) the appearance of anti-lymphocytic GM3 antibodies in asymptomatic anti-HIV+ subjects could have prognostic relevance for the risk of developing AIDS.
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Recent characterization of abnormal phosphatidylcholine metabolism in tumor cells by nuclear magnetic resonance (NMR) has identified novel fingerprints of tumor progression that are potentially useful as clinical diagnostic indicators. In... more
Recent characterization of abnormal phosphatidylcholine metabolism in tumor cells by nuclear magnetic resonance (NMR) has identified novel fingerprints of tumor progression that are potentially useful as clinical diagnostic indicators. In the present study, we analyzed the concentrations of phosphatidylcholine metabolites, activities of phosphocholine-producing enzymes, and uptake of [methyl-14C]choline in human epithelial ovarian carcinoma cell lines (EOC) compared with normal or immortalized ovary epithelial cells (EONT). Quantification of phosphatidylcholine metabolites contributing to the 1H NMR total choline resonance (3.20-3.24 ppm) revealed intracellular [phosphocholine] and [total choline] of 2.3 ± 0.9 and 5.2 ± 2.4 nmol/106 cells, respectively, with a glycerophosphocholine/phosphocholine ratio of 0.95 ± 0.93 in EONT cells; average [phosphocholine] was 3- to 8-fold higher in EOC cells (P < 0.0001), becoming the predominant phosphatidylcholine metabolite, whereas average g...
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INTRODUCTION Follicular lymphoma (FL) is considered a chronic, immune responsive and still incurable tumour. Recently two papers have shown the efficacy of active immunotherapy by repeated administration of autologous dendritic cells (DC)... more
INTRODUCTION Follicular lymphoma (FL) is considered a chronic, immune responsive and still incurable tumour. Recently two papers have shown the efficacy of active immunotherapy by repeated administration of autologous dendritic cells (DC) in Follicular Lymphoma (FL). Notably, in both studies a third of patients achieved long lasting remission. A novel DC population (IFN-DC), differentiated from human monocytes in the presence of GM-CSF and IFN-alpha has been developed in the laboratory of the Istituto Superiore di Sanità (ISS). IFN-DC have been demonstrated highly efficient in internalizing tumour-cell antigens, recovering…
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This pilot study explores the effects of a post-operative physical exercise program on the quality of life (QoL) and functional and nutritional parameters of patients that underwent laparoscopic colorectal cancer surgery, compared to... more
This pilot study explores the effects of a post-operative physical exercise program on the quality of life (QoL) and functional and nutritional parameters of patients that underwent laparoscopic colorectal cancer surgery, compared to usual care alone. The intervention group (IG) attended a 2-month-long supervised and combined exercise–training program during the post-operative period. Both IG and control group (CG) participated in the QoL, functional, and nutritional assessments before exercise training (T0), 2 months after the beginning of the exercise (end of treatment) (T1), and 2 (T2) and 4 (T3) months from the end of treatment. Eleven patients with colorectal cancer that underwent laparoscopic surgery were enrolled (six intervention; five control). The IG showed significant improvements compared to the CG in “Physical functioning” (PF2) (p = 0.030), “Cognitive functioning” (CF) (p = 0.018), and “Fatigue” (FA) (p = 0.017) of the European Organization for Research and Treatment o...
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Epstein-Barr virus-producing cells were used as a model to analyze, with a fracture-immunolabel technique, the distribution, behavior on fracture, and extent of glycosylation of viral transmembrane glycoproteins at the inner nuclear... more
Epstein-Barr virus-producing cells were used as a model to analyze, with a fracture-immunolabel technique, the distribution, behavior on fracture, and extent of glycosylation of viral transmembrane glycoproteins at the inner nuclear membrane. Surface and fracture immunolabeling with two monoclonal antibodies directed against the carbohydrate or polypeptide portions of the major viral envelope glycoproteins gp350/220 showed the following. (i) The glycoproteins present on the inner and outer nuclear membranes were labeled only with the monoclonal antibody directed against the polypeptide chain, whereas over the surface of virus-producing cells and on mature virions the labeling was dense and uniformly distributed with both monoclonal antibodies. (ii) The glycoproteins were nonuniformly distributed only over the inner nuclear membranes; at the sites of viral budding, the glycoproteins showed a preferential partition with the protoplasmic face. Since fully glycosylated glycoproteins wer...
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In this study we analyzed by immunofluorescence, laser confocal microscopy, immunoelectron microscopy and label fracture technique the ganglioside distribution on the plasma membrane of several different cell types: human peripheral blood... more
In this study we analyzed by immunofluorescence, laser confocal microscopy, immunoelectron microscopy and label fracture technique the ganglioside distribution on the plasma membrane of several different cell types: human peripheral blood lymphocytes (PBL), Molt-4 lymphoid cells, and NIH 3T3 fibroblasts, which mainly express monosialoganglioside GM3, and murine NS20Y neuroblastoma cells, which have been shown to express a high amount of monosialoganglioside GM2. Our observations showed an uneven distribution of both GM3 and GM2 on the plasma membrane of all cells, confirming the existence of ganglioside-enriched microdomains on the cell surface. Interestingly, in lymphoid cells the clustered immunolabeling appeared localized over both the microvillous and the nonvillous portions of the membrane. Similarly, in cells growing in monolayer, the clusters were distributed on both central and peripheral regions of the cell surface. Therefore, glycosphingolipid clusters do not appear confin...
Research Interests: Biology, Confocal Microscopy, Medicine, Signal Transduction, Humans, and 15 moreMice, Animals, Monoclonal Antibodies, Solubility, Peripheral blood lymphocyte, Fluorescent Antibody Technique, Monoclonal Antibody, Lymphocytes, Cell Surface Markers, Plasma Membrane, Polyethylene Glycols, Biochemistry and cell biology, Phosphate Buffer Saline, Cell Membrane, and Generating Function
Ovarian carcinomas represent a major form of gynaecological malignancies, whose treatment consists mainly of surgery and chemotherapy. Besides the difficulty of prognosis, therapy of ovarian carcinomas has reached scarce improvement, as a... more
Ovarian carcinomas represent a major form of gynaecological malignancies, whose treatment consists mainly of surgery and chemotherapy. Besides the difficulty of prognosis, therapy of ovarian carcinomas has reached scarce improvement, as a consequence of lack of efficacy and development of drug-resistance. The need of different biochemical and functional parameters has grown, in order to obtain a larger view on processes of biological and clinical significance. In this paper we report novel metabolic features detected in a series of different human ovary carcinoma lines, by (1)H NMR spectroscopy of intact cells and their extracts. Most importantly, a new ovarian adenocarcinoma line CABA I, showed strong signals in the spectral region between 3.5 and 4.0 p.p.m., assigned for the first time to the polyol sorbitol (39+/-11 nmol/10(6) cells). (13)C NMR analyses of these cells incubated with [1-(13)C]-D-glucose demonstrated labelled-sorbitol formation. The other ovarian carcinoma cell lin...
Research Interests: Hydrogen, British, Biology, NMR Spectroscopy, Magnetic Resonance Spectroscopy, and 15 moreOvarian Cancer, Medicine, Cell line, Humans, Female, Feature Detection, Drug Resistance, Nmr, Glutathione, Detoxification, Ovarian Carcinoma, Disease Progression, Sorbitol, Adenocarcinoma, and Ovarian Neoplasms
Research Interests: Prostate Cancer, Cell Division, Cell line, Humans, Collagen, and 13 moreClinical, Male, Plasminogen, Clinical Sciences, Proteoglycans, Neoplasm Invasiveness, Plasminogen Activator, Extracellular Matrix Proteins, Laminin, Prostate carcinoma, Prostatic neoplasms, Conditioned medium, and Drug combinations
The aim of this study is to investigate the effects in vitro induced by androgenic anabolic steroids (AAS) (testosterone, nandrolone, androstenedione, norandrostenedione, and norandrostenediol) used illicitly in sport competitions, on the... more
The aim of this study is to investigate the effects in vitro induced by androgenic anabolic steroids (AAS) (testosterone, nandrolone, androstenedione, norandrostenedione, and norandrostenediol) used illicitly in sport competitions, on the proliferation ability, apoptosis and the intracellular calcium concentration ([Ca2+]i) in human umbilical vein endothelial cells (HUVECs), selected as a prototype of a biological target system whose structure and function can be affected by steroids. For this purpose, we evaluated the proliferation inhibition by cytotoxic assay expressed as the concentration of drug inducing a 50% decrease in growth (IC50). The IC50 was reached for testosterone at 100 microM, androstenedione at 375 microM, nandrolone at 9 microM, norandrostenedione at 500 microM. The IC50 value for norandrostenediol was not reached until a concentration of 6000 microM. The apoptotic effect was evaluated by flow cytometry at IC50 for each drug. We observed that testosterone induced 31% of apoptotic cells, norandrostenedione 25%, androstenedione 15% and nandrolone 18%. We have analyzed the effects of these drugs on [Ca2+]i both in the immediate and long-term continuous presence of each compound. Our data show a statistically significant increase of [Ca2+]i in the acute condition and in long-term treated cultures, suggesting that androgen steroids modulate intracellular levels of calcium independent of incubation time or compound identity. As a whole, this study demonstrates that AAS might alter endothelial homeostasis, predisposing to the early endothelial cell activation that is responsible for vascular complications observed frequently in AAS users.
Research Interests: Flow Cytometry, Biology, Calcium, Apoptosis, STEROIDS, and 15 moreMedicine, Protein Structure and Function, Humans, Endothelial Cells, Statistical Significance, Endothelial cell, Cell Proliferation, Elsevier, Fluorometry, Doping in Sports, Intracellular Calcium, ENVIRONMENTAL SCIENCE AND MANAGEMENT, Androstenedione, Nandrolone, and Pharmacology and pharmaceutical sciences
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Research Interests: Biology, Ovarian Cancer, Medicine, Humans, Ph, and 15 moreVEGF, Female, Animals, Tumor Angiogenesis, Neoplasia, Fibroblast Growth Factor, Clinical Sciences, Cattle, Neoplasm Invasiveness, Endothelial cell, Culture Media, Biological Availability, Enzyme Linked Immunosorbent Assay, Cell Motility, and Ovarian Neoplasms
Research Interests: Mass Spectrometry, Biology, Biological Chemistry, Confocal Microscopy, Medicine, and 15 moreBiological Sciences, Humans, Biological, Gas Chromatography, High Pressure Liquid Chromatography, CHEMICAL SCIENCES, T lymphocytes, Fluorescent Antibody Technique, T cell activation, Thin Layer Chromatography, Plasma Membrane, Cell Membrane, Protein tyrosine kinases, Medical and Health Sciences, and human lymphocyte
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Research Interests: Fluorescence Microscopy, Biology, Scanning Electron Microscopy, Scanning Transmission Electron Microscopy, Confocal Microscopy, and 11 moreApoptosis, Medicine, Cell line, Humans, Quntitative Thin Layer Chromatography, Thin Layer Chromatography, Gangliosides, Biochemistry and cell biology, Fas, Gd, and Fas mediated apoptosis
Research Interests: Chemistry, Confocal Microscopy, Medicine, Cell line, Humans, and 15 morePeripheral blood lymphocyte, Gp, GM, Monoclonal Antibody, Quntitative Thin Layer Chromatography, Polyacrylamide Gel Electrophoresis, Protein Binding, Cell Surface Markers, Plasma Membrane, Biochemistry and cell biology, Phosphate Buffer Saline, Cell Membrane, Sodium Dodecyl Sulfate, Protein Kinase C, and Microdomain
OBJECTIVE: To assess possible differences in the activity of matrix metalloproteinases (MMPs), MMP-2 and MMP-9, and their inhibitors, the tissue inhibitors of MMPs, TIMP-1 and TIMP-2, in follicular fluid (FF) of women undergoing in vitro... more
OBJECTIVE: To assess possible differences in the activity of matrix metalloproteinases (MMPs), MMP-2 and MMP-9, and their inhibitors, the tissue inhibitors of MMPs, TIMP-1 and TIMP-2, in follicular fluid (FF) of women undergoing in vitro fertilization (IVF) treatment and of normally ovulating women. DESIGN: Prospective study. METHODS: MMP-2 and MMP-9 activity was analyzed by gelatin zymography and MMP-2, MMP-9, TIMP-2, TIMP-1 and 17beta-estradiol levels were measured in FF by ELISA. RESULTS: We found significantly reduced MMP levels in FF of women undergoing IVF treatment when compared with those of normally ovulating women. In contrast, the TIMP-1 levels were found significantly increased in FF from IVF patients vs normally ovulating women. No significant differences were found for TIMP-2 between the two groups. CONCLUSIONS: These findings underline a marked difference in MMPs and their inhibitors in the IVF women and the control group. Therefore we assume MMPs depend on hormonal s...
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PTX (Paclitaxel®) is an antimitotic agent used in the treatment of a number of major solid tumours, particularly in breast and ovarian cancer. This study was undertaken to gain insight into the molecular alterations producing PTX... more
PTX (Paclitaxel®) is an antimitotic agent used in the treatment of a number of major solid tumours, particularly in breast and ovarian cancer. This study was undertaken to gain insight into the molecular alterations producing PTX resistance in ovarian cancer. PTX treatment is able to induce apoptosis in the human ovarian carcinoma cell line, CABA I. PTX-induced apoptosis in CABA I cells was accompanied by an increase in the cellular Cer (ceramide) levels and a decrease in the sphingomyelin levels, due to the activation of sphingomyelinases. The inhibition of acid sphingomyelinase decreased PTX-induced apoptosis. Under the same experimental conditions, PTX had no effect on Cer and sphingomyelin levels in the stable PTX-resistant ovarian carcinoma cell line, CABA-PTX. The acquisition of the PTX-resistant phenotype is accompanied by unique alterations in the complex sphingolipid pattern found on lipid extraction. In the drug-resistant cell line, the levels of sphingomyelin and neutral ...
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Tumor cell ganglioside shedding has been implicated in the process of tumor formation. Previously, we identified three forms of tumor ganglioside shedding: micelles, monomers and membrane vesicles. Here, we have explored the membrane... more
Tumor cell ganglioside shedding has been implicated in the process of tumor formation. Previously, we identified three forms of tumor ganglioside shedding: micelles, monomers and membrane vesicles. Here, we have explored the membrane vesicle form of ganglioside shedding, using a newly identified human ovarian carcinoma cell line, CABA I. These cells synthesize and express a spectrum of gangliosides, including the disialoganglioside, G(D3). Immunostaining using the monoclonal antibody R24 confirmed G(D3) expression and its presence in the plasma membrane of these cells. Cellular gangliosides were detected in the culture supernatant by HPTLC autoradiography, confirming an active shedding rate of 3% of cellular gangliosides/24 h. CABA I cell membranes also express caveolin-1, a characteristic protein marker for caveolae, which was detected by flow cytometric analysis and by Western blotting in both the cell membranes and the isolated membrane vesicles. To further define the expression of G(D3) and caveolin-1, we used immunogold electron microscopy. This revealed localization of G(D3) in small clusters in the plasma membrane as well as enrichment and localization of ganglioside G(D3) and caveolin-1 in shed membrane vesicles, with 58-78% of vesicles carrying both G(D3) and caveolin-1. Together, these results suggest that membrane vesicle shedding originates in plasma membrane domains enriched in gangliosides and caveolin-1.
Research Interests: Electron Microscopy, Flow Cytometry, Membrane Proteins, Immunohistochemistry, Biological Sciences, and 14 moreCell line, Humans, Female, Physical sciences, Spectrum, Western blot, Ovarian Carcinoma, Monoclonal Antibody, Caveolin-1, Plasma Membrane, Gangliosides, Cell Membrane, Medical and Health Sciences, and Ovarian Neoplasms
Research Interests: Immunology, Stem Cells, Adolescent, Cell Differentiation, Humans, and 15 moreEndothelial Cells, Systemic sclerosis, Female, Male, Mesenchymal Stem Cell, Arthritis, Phenotype, Clinical Sciences, Middle Aged, Adult, Endothelial cell, Public health systems and services research, Immunophenotyping, Mesenchymal Stromal Cells, and Case Control Studies
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The aim of this study was to further elucidate our previous observation on molecular interaction of GM3, CD4 and p56lck in microdomains of human peripheral blood lymphocytes (PBL). We analyzed GM3 distribution by immunoelectron microscopy... more
The aim of this study was to further elucidate our previous observation on molecular interaction of GM3, CD4 and p56lck in microdomains of human peripheral blood lymphocytes (PBL). We analyzed GM3 distribution by immunoelectron microscopy and the association between GM3 and CD4-p56lck complex by scanning confocal microscopy and co-immunoprecipitation experiments. Scanning confocal microscopy analysis showed an uneven signal distribution of