A R Mariani

University of Bologna, Bolonia, Emilia-Romagna, Italy

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Publications (22)42.94 Total impact

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    ABSTRACT: Molt-4 human leukemia cells were triggered to apoptosis by various agents with different mechanisms of action. Staurosporine, a protein kinase C (PKC) inhibitor; camptothecin, a topoisomerase I blocking drug; and tiazofurin, an inhibitor of inosine 5'-phosphate dehydrogenase (IMPDH), were used. Ultrastructural analysis showed morphologic changes characteristic of apoptosis that were very similar for all three agents. Nevertheless, DNA oligonucleosomic fragmentation was not detectable by agarose gel electrophoresis. However, a genomic DNA cleavage appeared after pulse-field gel electrophoresis (PFGE) in cells treated with these agents for 24 h. Furthermore, in situ nick translation (NT) showed a finely spotted nuclear labelling in staurosporine-treated cells and a compact fluorescence after camptothecin incubation. In tiazofurin-treated cells an intermediate pattern was found. Therefore, apoptotic agents with different mechanisms of action induced the formation of large genomic DNA fragments and very similar ultrastructural changes. Therefore, both phenomena and the closely related apoptosis progression depend on target cell machinery and not on the triggering agent.
    No preview · Article · Dec 2006 · Scanning
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    ABSTRACT: Insertion tissue biopsies of right arm common extensor tendons from 11 patients with chronic lateral epicondylitis were processed for light and electron microscopy. The subjects were aged between 38 and 54 years (only one was 25). The specimens showed a variety of structural changes such as biochemical and spatial alteration of collagen, hyaline degeneration, loss of tenocytes, fibrocartilage metaplasia, calcifying processes, neovascularization and vessel wall modifications. Tissue alterations were evident in limited zones of the tendon fibrocartilage in which the surgical resection was generally visible. The areas where the degenerative processes were localized, were restricted and in spatial contiguity with morphologically normal ones. The observed cases presented histological and electron microscopic findings that characterize lateral epicondylitis as a degenerative phenomenon involving all tendon components.
    Preview · Article · Feb 2002 · European journal of histochemistry: EJH

  • No preview · Article · Sep 2001 · The FASEB Journal
  • L. Zamai · A.R. Mariani · S. Papa · F. Reno · M. Vitale
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    ABSTRACT: Background. Natural Killer cells are an important line of defense against newly arising malignant cells and cells infected with viruses, bacteria, and protozoa. They have the ability to mediate a spontaneous cytotoxicity against a variety of cell types including normal allogeneic cells, virally infected cells, and tumor cells. NK cell activity is finely tuned by the opposing activity of stimulatory and inhibitory receptors, with specificity for various MHC-I allotypes. Lysis of the target can be achieved by different mechanisms, including cytotoxic granule exocytosis and production of soluble factors. We studied the possibility offered by flow cytometry to give a comprehensive quantitative evaluation of human NK cell activity. Method. Human MC cells, recognised by specific monoclonal antibodies, and tumor targets were analysed by flow cytometry after the binding and killing phases of their activity. Results. The two phases of NK lytic activity (binding and killing) can be measured by flow cytometry, in a single test tube. The flow cytometry methodologies described here give the quantitative measurement of NK cell binding to tumor targets and of their lytic activity. Conclusions. The measurement of NK cell function should be performed in specific instances, such as on patients experiencing or suspected of autoimmune disorders, chronic fatigue immune dysfunction syndrome (CFIDS), repeated infections, chemotherapy, treatment with biological modifiers such as IL-2. Flow cytometry can measure both binding and killing NK cell functions, and is fast, independent of the operator, statistically reliable, avoids the use of radioactive material and performs the analysis at single cell level rather than on whole PBL.
    No preview · Article · Jun 1999 · Minerva Biotecnologica
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    E Mariani · A R Mariani · A Meneghetti · A Tarozzi · L Cocco · A Facchini
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    ABSTRACT: The progressive increase in the number of peripheral NK cells found in the elderly does not correlate with a corresponding increase in lytic activity. On the contrary, a decreased function of circulating NK cells purified from old subjects was observed on a per cell basis. Most of the studies on NK cells have focused on late events such as lytic activity. In view of this, little is currently known about the modification of the early signalling pathways of NK cells in elderly people. This study investigated whether the modification of NK lytic activity could be related to differences in the metabolic pattern of activation of these cells in the elderly. NK cells were negatively purified by immunomagnetic depletion from the peripheral blood of selected old and young healthy subjects. Hydrolysis of inositol phospholipids was measured following incubation with K562 target cells and/or CD16 mAb for different times. Our data show that there is a pronounced age-related decrease in the ability to generate total inositol monophosphates and, particularly, inositol trisphosphates by NK cells following K562 stimulation (spontaneous cytolytic activity) together with an attenuated and delayed hydrolysis of phosphatidylinositol bisphosphate, while phosphoinositide turnover is preserved following Fc triggering (antibody-dependent cell-mediated cytotoxicity). These results confirm that, also in old subjects, different biochemical pathways of activation are involved in NK cells when target or antibody-mediated triggering occurs and may aid the development of experimental and therapeutic strategies to counteract declines in cell mediated immune functions associated to advancing age.
    Full-text · Article · Aug 1998 · International Immunology
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    L Rodella · R Rezzani · G Zauli · A R Mariani · R Rizzoli · M Vitale
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    ABSTRACT: NK cells kill sensitive targets via exocytosis of cytoplasmic granules containing membrane damaging perforins and DNA damaging granzymes. Therefore, the target cell can either die by necrosis or by apoptosis. A third, non-secretory, mechanism of killing mediated by Fas-FasL interaction can be used by activated NK cells to kill Fas+ targets. Here, we have studied the modulation exerted by two NK active cytokines, IL-2 and IL-12, on the apoptosis-inducing activity of NK cells in NK-sensitive Fas- K562 and NK-insensitive Fas+ Raji cells. Our results show that apoptosis is preferentially induced at low target:effector ratios. Resting NK cells virtually do not induce apoptosis in target cells while IL-2 stimulation endows NK cells with the capacity of inducing apoptosis and IL-12 further enhances this effect against both targets. Finally, we demonstrate that cytokine-stimulated NK cells use both granzyme and Fas-FasL pathways of apoptosis induction.
    Full-text · Article · Jul 1998 · International Immunology
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    ABSTRACT: The morphological features of cell undergoing programmed cell death is well known and has been widely described in a number of experimental models with a variety of apoptotic triggering agents. Despite the similar cell behaviour, underlying molecular events seem variable and only partially understood. A multiple approach appears crucial to better clarify the phenomenon. The first technique, DNA gel electrophoresis, allows the identification of fragmented DNA and has been long considered the hallmark of apoptosis. Different patterns of DNA cleavage, which can be identified by conventional or "pulsed-field gel" electrophoresis, are presented and discussed. "In situ" labelling methods are also described both with terminal deoxynucleotidyl transferase and DNA polymerase I, aimed at the study of the distribution of DNA cleavage areas. Flow cytometry is also proposed and different technical approaches, based on different laser utilizations, are discussed. Ultrastructural analysis, allowing the study of apoptotic cell details, is finally considered.
    No preview · Article · Feb 1996 · Scanning microscopy
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    ABSTRACT: Cytotoxicity assays are widely used to evaluate the functional activity of NK and T cells against tumour target cells and the release of radioactive sodium chromate from labelled target cells is still the most commonly used marker of target lysis in culture supernatants. We describe here the standardization of a micro-cytotoxicity test in which the number of cytolytic effector and tumour target cells have been decreased by a factor of 10. The release obtained by 500 tumour target cells was compared with the release obtained by 5000 target cells in the standard cytotoxicity assay for target:effector cell ratios from 1:1 to 1:100. Both gamma and beta emissions of the 51Cr isotope were evaluated to determine the assay release. The results obtained by the micro-cytotoxicity assay (500 target cells) were comparable to those of the standard assay (5000 target cells) and 51Cr release evaluation using the gamma counter was the most sensitive method of determining lytic activity using 500 tumour target cells. beta counter evaluation using solid phase scintillation was found to be a reproducible alternative method, even if the lytic curves cannot be compared with those obtained using the traditional method.
    Full-text · Article · Jul 1994 · Journal of Immunological Methods
  • E Mariani · A.R. Mariani · M.C.G. Monaco · E Lalli · M Vitale · A Facchini
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    ABSTRACT: Various growth factors, hormones and proteins present in serum are presumed to be responsible for its growth-stimulating activity in culture media. However, synthetic or serum substitute media supporting cell growth are advantageous when it is necessary to standardize culture conditions, particularly when cell products are used. In this study we have evaluated and compared the effects of some commercially available serum substitute media (10% NU Serum, 10% BMS, 2% Ultroser HY, 1% ITS + Premix, 1% Nutridoma, Ultradoma, FEB100, DCCM1 and DCCM2) on growth and immunoglobulin production in different hybridoma cell lines. Six different hybrids were studied: OKT3, OKT4 and OKT8 (producing monoclonal antibodies against CD3, CD4 and CD8 molecules), HB43 and HB57 (producing monoclonal antibodies against human IgG and IgM), and CRL 8019 (producing monoclonal antibodies against high-molecular weight CEA). Several parameters were evaluated, such as viability, doubling time, DNA synthesis, monoclonal antibody production and cell cycle under different culture conditions. Since not all of the hybridomas grew equally well in the same serum substitute media, one synthetic medium cannot be used for all the lines. We found that the serum-free media that best supported hybridoma growth were Nutridoma, DCCM1, DCCM2, NU Serum and FEB100.
    No preview · Article · Jan 1992 · Journal of Immunological Methods
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    E Mariani · P Roda · A R Mariani · M Vitale · A Degrassi · S. and Papa · A Facchini
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    ABSTRACT: A cloning technique was used to estimate the frequency of proliferating T cell precursors, the growth capacity of clone-forming cells and the functional activity of clones established in vitro from peripheral blood lymphocytes of young and old people. The mean frequency of proliferating precursors was lower in the elderly as was the proliferative capacity of CD8+ clones. In contrast, CD4+ and CD16+ clones showed a proliferation similar to that obtained from young subjects. When the clones were examined for their functional activity, CD4+ clones from both groups failed to show any cytolytic activity, while CD8+ clones exerted cytolysis against K562 and in antibody-dependent cell-mediated cytotoxicity but this function was reduced in clones derived from old subjects. Similarly, CD16+ clones from the elderly showed a decreased activity at some effector-to-target cell ratios. We conclude that the impaired functional activity (T or NK-dependent) found in the peripheral blood of aged subjects persists after in vitro selection when these cells are analysed at clonal level.
    Full-text · Article · Oct 1990 · Clinical & Experimental Immunology
  • E Falcieri · A R Mariani · E Mariani · P Gobbi · A Facchini · F A Manzoli
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    ABSTRACT: Early and late events occurring during cell-mediated cytolysis have been examined ultrastructurally using transmission, scanning and freeze-fracture electron microscopy. K562 erythroleukemic cells were used as target for human natural killer lymphocytes, a subpopulation involved in host defense against tumor, virus infected and allogeneic cells. Cell binding between target and effector is already evident after 15 min, but membrane lesions and target hydration appear after 30 min incubation. At 60 min K562 cells appear swollen and microvilli-deprived. Progressive membrane disruption is also evident and the underlying cytoplasmic material is clearly exposed after 120 min. Submicroscopic membrane lesions appear to be early events in comparison with 51Cr (isotope) release from damage cells.
    No preview · Article · May 1990 · Journal of submicroscopic cytology and pathology
  • E Falcieri · A R Mariani · R Del Coco · A Facchini · N M Maraldi
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    ABSTRACT: A simple, high-yield technique for the freeze-fracturing of small amounts of isolated cells is described. A drop of cells fixed in suspension is deposited on a polylysine-treated coverslip, forming a monolayer through electrostatic forces. After cryoprotection, the coverslip is inverted on a gold carrier covered with Vinol and then frozen in liquid nitrogen. The monolayer will be fractured by advancing the knife under the coverslip. Large areas of cell surface can be exposed despite their low number, such as that obtainable after cell sorting by flow cytometry.
    No preview · Article · Aug 1988 · Journal of submicroscopic cytology and pathology
  • S Papa · M Vitale · A R Mariani · P Roda · A Facchini · F A Manzoli
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    ABSTRACT: Flow cytometry has been employed to establish an NK assay using the K562 target cell line. These cells show a perpendicular light scatter (PLS) characteristically different from lymphocytes. Other physical parameters, such as forward light scatter (FLS), do not discriminate between the two populations, since dead K562 cells display similar FLS characteristics as effector cells. Killed cells are stained with propidium iodide and followed by flow analysis. It was advisable to add the dye in the medium so that, as long as the target cells are killed they will also be stained. Moreover the flow cytometric and trypan blue evaluation of target cell death rate showed a stronger correlation than did either test with the conventional 51Cr release assay, the first two methods both being based on the same biological mechanism.
    No preview · Article · Mar 1988 · Journal of Immunological Methods
  • E Mariani · P Roda · A R Mariani · M Vitale · A Degrassi · S Papa · F A Manzoli · A Facchini
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    ABSTRACT: Old subjects exhibit a decline in circulating T cells and an impaired proliferative response to mitogens, plus a relative increase in cells with NK phenotype not associated with a concomitant increase in their cytolitic activity.In the present study a limiting dilution assay was used to evaluate the phenotype, the functional activity and the proliferative capacity of clones obtained from peripheral blood lymphocytes of old and young subjects. CD5+ CD8+ clones from old people showed a significant impairment in their proliferative capacity and a decreased lytic activity against K562 and P815-IgG cell lines.
    No preview · Article · Oct 1987 · Cytotechnology
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    A Facchini · E Mariani · A R Mariani · S Papa · M Vitale · F A Manzoli
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    ABSTRACT: The phenotype and functional activity of circulating T and natural killer (NK) cells was investigated in a group of selected normal elderly donors in comparison with a group of normal young individuals. Old subjects exhibit a decline in circulating T cells and an impaired response to mitogens (Con A, PHA) associated with a relative increase in cells reacting with Leu 7 and Leu 11a monoclonal antibodies, directed against large granular lymphocytes with NK activity. The rise in circulating NK cells observed in the elderly was not associated with a concomitant increase in NK cytolytic activity against K 562 tumour cells, and time course kinetics of the cytotoxic reaction was similar to that observed in young subjects. When a greater than 95% pure population of Leu 11a+ cells from old individuals was sorted by flow cytometry and their functional cytotoxic activity examined, a significant decrease in NK cytotoxicity was detected. The target cell binding capacity of effector cells, morphologically evaluated at single cell level, did not differ in old and young subjects, even though bound cells from young donors have a higher lytic capacity. These data show that an increase in circulating NK granular lymphocytes occurs during human ageing, but suggest that in old subjects only a subset of NK cells is active in order to maintain a functional response similar to that observed in the young.
    Full-text · Article · Jun 1987 · Clinical & Experimental Immunology
  • A Facchini · E Mariani · A Ferrolli · A R Mariani · M Gobbi · F Zizzi · L Frizziero

    No preview · Article · Jan 1982 · Arthritis & Rheumatology

  • No preview · Article · Dec 1981 · Arthritis & Rheumatology
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    ABSTRACT: The incidence of LMA, detected by indirect IF on isolated rabbit hepatocytes, was significantly higher in CAH-non B (58%) than in CAH-B (15%). Sera from AVH patients showed a smaller percentage of positivity whereas the CPH patients were negative. The organ-specificity, the high incidence in relation to other non-organ-specific autoantibodies and the inverse correlation with the markers of HB Virus, make LMA a specific and sensitive marker of "autoimmune" chronic liver disease.
    No preview · Article · Sep 1980 · Bollettino della Società italiana di biologia sperimentale
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    ABSTRACT: During the last few years a number of experimental evidences have shown the presence of Fc receptors for IgG or IgM on the membrane of human T cells. These two different receptors are detectable and mutually exclusive on distinct cell populations named respectively TG, TM and T "null" (which lack detectable receptors). Studies on the functional activities of these cells have shown that TM and TG lymphocytes play an antitetical role in regulating B cell response, TM exerting an "helper" activity on the differentiation of B lymphocytes while TG having a "suppressor" one. The aim of this study has been to determine the values of these two subpopulations in a group of twenty control subjects. Our results have shown that TG constitute 10%, whereas TM represent 40% of the total T cells. After EA-G rosetting, the purification of this subpopulation on a density gradient has shown an enrichment of more than 90% in TG cells, while TM contaminate this fraction for less than 4%. The purity of the fraction containing TM has been evaluated using the localization of alpha-naphthyl acetate esterase activity, which has shown that more than 88% of the cells in this fraction are positive for this enzyme.
    No preview · Article · Sep 1980 · Bollettino della Società italiana di biologia sperimentale
  • G Ramadori · E Mariani · A Facchini · A Mariani · R Giammarco

    No preview · Article · Sep 1977 · Bollettino della Società italiana di biologia sperimentale

Publication Stats

391 Citations
42.94 Total Impact Points


  • 1981-2006
    • University of Bologna
      • • Department of Biomedical Science and Neuromotor Sciences DIBINEM
      • • Institute of Genetic Medicine
      Bolonia, Emilia-Romagna, Italy
  • 2001
    • Università degli studi di Parma
      • Department of Clinical and Experimental Medicine
      Parma, Emilia-Romagna, Italy
  • 1999
    • Università degli Studi di Urbino "Carlo Bo"
      Urbino, The Marches, Italy
  • 1998
    • Istituto Ortopedico Rizzoli
      Bolonia, Emilia-Romagna, Italy