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L Zamai

Publications and source records attributed to L Zamai.

46 records · Page 3Linked to original sources
PubMed1992

The impairment of natural killer function in the healthy aged is due to a postbinding deficient mechanism.

In order to study the fine mechanisms that underlie the impairment of non-MHC-restricted cytolytic activity which occurs during human aging, we examined by multiparametric flow cytometry the binding and lytic activities of human natural killer cells. The flow analysis revealed a striking increase of the CD16+8- subset, together with a significant decrease of CD8bright cells and total T cells (CD3+). Aging had no influence on the CD8dim subset. The total lytic activity expressed by PBL as well as their binding efficiency to K562 targets were moderately but not significantly increased in the elderly. In contrast, the cytotoxicity of the single target-bound natural killer cell (i.e., lytic efficiency) was deeply impaired in aged subjects, suggesting that the NK functional impairment observed in aging is located at postbinding level.

Adult↗
PubMed1992

Interferon affects cell growth progression by modulating DNA polymerases activity.

A multiparametric analysis of the effects of human recombinant interferon alpha type A on Daudi cells involving flow cytometry and in vitro analysis of alpha and beta DNA polymerase activities has been performed. Results have disclosed (within 60 min of interferon treatment) a decrease of alpha polymerase driven DNA synthesis persisting to at least 24 h, while beta polymerase was poorly affected. Moreover, after 24 h of interferon treatment, a reduction of BrdUrd incorporation per cell, assessed by flow cytometry, was observed suggesting that DNA synthesis in S phase cells is almost completely abolished. The analysis of the effect of interferon on the distribution of cell cycle phases indicated that the G1/S transition is not inhibited by the treatment. These results support the hypothesis that interferon generates a transient initiating signal which quickly reaches the nucleus and produces a rapid inhibition of alpha polymerase activity, leading finally to the slowing of cell cycle progression.

Cell Cycle↗
PubMed1991

Natural killer function in flow cytometry: identification of human lymphoid subsets able to bind to the NK sensitive target K562.

NK cells are a phenotypically, morphologically and functionally heterogeneous population. This has led to the current thought that the non-MHC restricted cytotoxicity is a cellular function that can be associated to different phenotypes. The recognition of the target cell and the conjugate formation is always the first step that eventually leads to the lysis of target. Characterization of the phenotypical pattern of the cells able to bind to K562 targets is the purpose of this study. A multi-parametric flow cytometry binding assay has been employed to identify the different K562-bound lymphocyte subsets. In particular, cells that coexpress the CD16 and CD8 antigens (CD16+8dim+) showed a significantly higher binding capacity than their CD16+8- counterpart. Moreover, the highest binding values have been found in cells that did not express the CD16 antigen at all, but still expressed the CD8dim antigen, such as the small CD8dim+3+ population. These data show that, the NK lytic function being dependent on binding, minor subpopulations must be considered among effector cells, which might correspond to different lytic activities. None of the previously published methodologies that analyze conjugates by flow cytometry or fluorescence microscopy were able to measure the binding capacity of small, double stained, lymphocyte subsets.

Adult↗
PubMed1991

Technical improvements for flow karyotyping by standard FACS 440 flow cytometer.

Chromosomal DNA measurements represent the most accurate methodology for the quantitative analysis of human karyotype. A modified FACS 440 signal collection is described here. The FSC signal voltage coming from the optical bench has been split in two equal parts by a T-connector placed at the preamplifier entrance. The side scatter channel that is not employed during chromosome analysis has been used for the half-FSC signal. This modification allowed the contemporary use of FSC signal both for gating and analysis leading to a better discrimination of background, and therefore to a more accurate definition of sorting windows.

Cell Line↗
PubMed1991

Nuclear metabolic changes induced by tumor necrosis factor in Daudi lymphoma cells; a multiparametric analysis.

The effects of r-TNF alpha on cell cycle progression and DNA polymerase activity in Daudi lymphoma cells have been analyzed. Cytofluorimetric analysis of the cell cycle after 6 to 24 hr of treatment revealed both a decrease of BrdU incorporation per cell and a light inhibition of S phase as assessed by the analysis of the percentual distribution of cell cycle compartments. The reduction of BrdU incorporation can be related to the early decrease in the rate of DNA synthesis that follows r-TNF alpha treatment. These results suggest that one of the early events induced by r-TNF alpha at nuclear level is the slowering of DNA synthesis leading to a reduced cell cycle progression.

Bromodeoxyuridine↗
PubMed1991

Characterization and cell cycle kinetics of hepatocytes during rat liver regeneration: in vivo BrdUrd incorporation analysed by flow cytometry and electron microscopy.

The incorporation of bromodeoxyuridine (BrdUrd) into newly synthesized DNA has been analysed during hepatocellular regeneration induced by partial hepatectomy in young rats. The kinetic state of the liver has been studied by flow cytometric analysis of the incorporated BrdUrd, while the fine localization of DNA replication sites through the cell cycle has been investigated at the ultrastructural level by the immunogold technique. Eighteen hours after partial hepatectomy flow cytometry revealed an early S phase distribution which corresponded to a specific staining of the interchromatin domains of the hepatocyte nucleus. Thirty-four hours after hepatectomy, on the other hand, when most cells were in late S, a specific staining of heterochromatin domains was observed. The effect of the BrdUrd technique on nuclear aggregation has also been analysed and discussed. The results demonstrate that specific patterns of DNA replication can be recognized during the cell cycle and that flow cytometry and electron microscopy appear to be complementary in the kinetic study of liver regeneration.

Animals↗
PubMed1991

Cell cycle analysis in flow cytometry: use of BrdU labelling and side scatter for the detection of the different cell cycle phases.

Cell cycle analysis in flow cytometry is based on the incorporation of labelled precursors in DNA. The use of BrdU versus SSC, in which side scatter substitutes PI fluorescence, has proved to be useful also for the distinction between G2 and Mitotic cells. Mitoses often produce an SSC decrease due to the morphological changes that happen in the nucleus during this phase of cell cycle. Moreover, DNA accessibility to PI varies during mitosis, as well. However, most of these variations, detectable by flow cytometry appear to be basically dependent on the cell line used.

Bromodeoxyuridine↗
PubMedDate not supplied

CD135.

Explore the source record for details and available documents.

Amino Acid Sequence↗
PubMedDate not supplied

Intracellular detection of Bcl-2 and p53 proteins by flow cytometry: comparison of monoclonal antibodies and sample preparation protocols.

Several techniques have been proposed for flow cytometric evaluation of intracellular antigens. This approach is particularly important for detection at the single cell level of proteins which correlate to tumour progression. Bcl-2 and p53 are two of the most relevant proteins. In the present study we have compared five different cell fixation-permeabilisation protocols and nine fluorochrome-conjugated (FITC or PE) monoclonal antibodies (mAb): four mAb directed against Bcl-2 and five against p53. For detection of Bcl-2 we have analysed three Bcl-2 positive cell lines (K562, Daudi and MCF-7), and peripheral blood samples obtained from nine healthy subjects. To distinguish internal positive (lymphocytes) and negative control cells (granulocytes), it was necessary to perform simultaneous detection of surface and intracellular antigens. For detection of p53 three cell lines, two p53 positive (Raji and CEM) and one p53 negative (HL-60), were analysed. Using these cells we have performed a combined analysis of the efficiency of monoclonal antibodies and sample preparation techniques. In conclusion, clones 124-FITC and Bcl-2/100-PE (Bcl-2), and clones BP53,12-FITC and G59-12-PE (p53) provided the highest specific fluorescence intensity of the respective markers independent of cell preparation protocols. Importantly, our results show that mAb background may depend on the specific fixation/permeabilisation kit and that mAb titration using negative and positive control cells is essential to determine the specificity and the sensitivity of the mAb used.

Antibodies, Monoclonal↗