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S Bertone

Publications and source records attributed to S Bertone.

At least 19 recordsLinked to original sources

Freak waves in random oceanic sea states.

Freak waves are very large, rare events in a random ocean wave train. Here we study their generation in a random sea state characterized by the Joint North Sea Wave Project spectrum. We assume, to cubic order in nonlinearity, that the wave dynamics are governed by the nonlinear Schrödinger (NLS) equation. We show from extensive numerical simulations of the NLS equation how freak waves in a random sea state are more likely to occur for large values of the Phillips parameter alpha and the enhancement coefficient gamma. Comparison with linear simulations is also reported.

Journal Article↗

Artificial neural network based identification of environmental bacteria by gas-chromatographic and electrophoretic data.

Chemotaxonomic identification techniques are powerful tools for environmental micro-organisms, for which poor diagnostic schemes are available. Whole cellular fatty acid methyl esters (FAME) content is a stable bacterial profile, the analysis method is rapid, cheap, simple to perform and highly automated. Whole-cell protein is an even more powerful tool because it yields information at or below the species level. The description of new species and genera and subsequent continuous rearrangement provide large amounts of data, resulting in large databases. In order to set up suitable software tools to work on such large databases artificial neural network (ANN) based programs have been used to classify and identify marine bacteria at genus and species levels, starting from the fatty acid profiles and protein profiles respectively. We analysed 50 certified strains belonging to Halomonas, Marinomonas, Marinospirillum, Oceanospirillum and Pseudoalteromonas genera. Both supervised and unsupervised ANNs provide a correct classification of the marine strains analyzed. Moreover, a set of 73 marine fresh isolates were used as an example of identification using ANNs. We propose supervised and unsupervised ANNs as a reliable tool for classification of bacteria by means of their FAME and of whole-protein analyses and as a sound basis for a comprehensive artificial intelligence based system for polyphasic taxonomy.

Bacteria↗

Application of artificial neural network for the identification of fresh water bacteria.

A method based on artificial neural network (ANN) for monitoring aquatic bacteria which would be useful for health care is presented. Environmental micro-organisms include a large number of taxa. Some species that normally are not pathogenic can represent a risk in certain conditions, such as old people and immuno-compromised individuals. A system based on unsupervised ANN has been set up using the fatty acid profiles of standard strains, obtained by gas-chromatography, as learning data. The Kohonen output map resulted in a powerful tool for identification of fresh isolates coming from a line of the major civil water system of Genova (Italy).

Bacteria↗

Inhibitory receptors sensing HLA-G1 molecules in pregnancy: decidua-associated natural killer cells express LIR-1 and CD94/NKG2A and acquire p49, an HLA-G1-specific receptor.

Trophoblastic cells lack classical HLA class I and class II molecules but express HLA-G1. Although this may prevent allorecognition by maternal T cells, it renders trophoblastic cells potentially susceptible to lysis by natural killer (NK) cells. As shown here, only a fraction of peripheral-blood NK cells in pregnant women express the HLA-G1-specific CD94/NKG2A and/or LIR-1 receptors. However, all NK cells isolated from maternal decidua during the first trimester expressed either one or both of these receptors. Perhaps more importantly, a fraction of cells expressed p49, an HLA-G1-specific inhibitory receptor, undetectable in peripheral-blood NK cells. p49 was expressed on virtually all NK cells isolated from placenta at term. Functional analyses revealed that the HLA class I-negative 221 lymphoblastoid cell line transfected with HLA-G1 was only partially protected from lysis by peripheral-blood NK cells isolated from pregnant women, whereas it was fully protected from decidual NK cells. As indicated by the addition of specific antibodies to cytolytic tests, all the above receptors contributed to HLA-G1 recognition by decidual NK cells, although p49 would appear to play a predominant role.

Animals↗

Transforming growth factor-beta-induced expression of CD94/NKG2A inhibitory receptors in human T lymphocytes.

Different HLA class I-specific killer inhibitory receptors (KIR) are expressed in vivo by a fraction of activated T cells, predominantly CD8+, in which they may inhibit TCR-mediated cell functions. In an attempt to identify mechanisms leading to KIR expression in T cells, we analyzed the effect of transforming growth factor-beta (TGF-beta) in T cells responding to bacterial superantigens in vitro. We show that TGF-beta induces the expression of CD94/NKG2A in cells responding to toxic shock syndrome toxin 1 or to other staphylococcal superantigens. Remarkably, maximal CD94 expression occurred at (low) TGF-beta concentrations which have no substantial effect on lymphocyte proliferation. Maximal CD94 expression occurred when TGF-beta was added shortly after the cells were placed in culture. No expression could be induced in CD94/NKG2A-negative T cell clones. Although both CD4+ and CD8+ expressed CD94, the simultaneous expression of NKG2A was mostly confined to CD8+ cells. Monoclonal antibody-mediated cross-linking of CD94/NKG2A led to an impairment of T cell triggering via CD3, as determined in a redirected killing assay using the Fcgamma receptor-positive P815 murine target cells.

Animals↗

An integration system to rapidly identify aquatic bacteria.

An identification system for aquatic bacteria which would be useful for aspects ranging from pollution monitoring to health care, is described. It contains relevant information concerning the nature, taxonomy and activity of aquatic bacteria and allows to identify fresh isolates by means of their chemotaxonomic profiles. The system is simple and user friendly, so that it can be used by people not familiar with computers and has a modular interface that allows an easy interaction with laboratory instruments. The system is also connected with artificial neural network based programs to identify strains starting from the considered chemotaxonomic features.

Bacteria↗

HLA class I-specific inhibitory receptors in human T lymphocytes: interleukin 15-induced expression of CD94/NKG2A in superantigen- or alloantigen-activated CD8+ T cells.

A fraction of human T lymphocytes, predominantly CD8+, express receptors for HLA class I molecules typical of natural killer cells (natural killer receptors or NKRs) that inhibit T cell receptor-mediated functions. Herein, we analyzed possible mechanism(s) leading to the expression of NKRs by T cells responding to superantigens or allogeneic cells in vitro. We show that, in the presence of interleukin 15 (IL-15), T cells (depleted of NKR+ cells) responding to toxic shock syndrome toxin 1 de novo express CD94, a molecule that is part of a heterodimeric NKR with a broad specificity for different HLA class I alleles. Maximal CD94 expression occurred when IL-15 was added shortly after the cells were placed into culture, and CD94 expression started 4-6 days after addition of IL-15. Although both CD4+ and CD8+ cells expressed CD94, the simultaneous expression of NKG2A (i.e., the other component of the CD94/NKG2A inhibitory NKR) was confined to CD8+ cells. Similar data were obtained in T cell populations activated in mixed lymphocyte cultures in the presence of IL-15. The expression of CD94/NKG2A led to an impairment of allo-specific cytolytic activity by mixed lymphocyte culture-derived T cell populations or clones. Remarkably, cytolysis could be restored by the addition of anti-CD94 mAb, i.e., by masking the inhibitory NKRs.

Antigens, CD↗

Cytokine-induced expression of killer inhibitory receptors in human T lymphocytes.

Killer inhibitory receptors (KIRs) represent a new family of HLA-class I-specific receptors. KIRs are involved in the function of Natural Killer cells and allow these cells to discriminate between normal cells and cells with impaired expression of HLA-class I molecules. KIRs are also expressed by a subset of cytolytic T lymphocytes in which they may exert an inhibitory effect on TCR-mediated function. Here we review recent data indicating that cytokines such as IL-15, may induce the de novo expression of CD94/NKG2A (a KIR which operationally detects the expression of various HLA-class I alleles). The expression of CD94/NKG2A has been documented not only in CD34+ precursors undergoing maturation towards NK cells, but also in mature T cells which respond in vitro to superantigens or allogeneic cells.

Animals↗

Interleukin-15-induced maturation of human natural killer cells from early thymic precursors: selective expression of CD94/NKG2-A as the only HLA class I-specific inhibitory receptor.

Immature postnatal thymocytes were shown to contain precursors which, under suitable culture conditions, give rise to phenotypically and functionally mature natural killer (NK) cells. Here, we analyzed the effect of different cytokines for their ability to induce the expression of HLA class I-specific inhibitory receptor(s) during the process of NK cell development from immature thymocytes. From thymocyte cell suspensions depleted of CD2+, CD3+, CD4+, CD8+, CD56+, and CD16+ cells, we further removed cells expressing HLA class I-specific inhibitory receptors including CD94/NKG2-A, p58.1, and p58.2 by immunomagnetic bead separation. The resulting cells did not contain any of the above NK receptors as determined by immunofluorescence and flow cytometric analysis, as well as by reverse transcriptase polymerase chain reaction (RT-PCR) amplification using appropriate sets of primers. Although different cytokines have been used, including interleukin (IL)-7, stem cell factor (SCF), IL-2, and IL-15, only IL-2 or IL-15 induced cell proliferation when used alone. Moreover, maturation towards CD3- CD56+ cells displaying cytolytic activity against the HLA class I- targets K562 or 221 was detectable in cultures containing IL-15 used alone or in combination with IL-7 or SCF. On the other hand, these CD3- CD56+ cell populations did not lyse HLA class I+ target cells, including autologous PHA blasts. Analysis of the expression of the various HLA class I-specific inhibitory NK receptors revealed the presence of high proportions of CD94/ NKG2-A+ cells, while the NK receptors belonging to the Ig superfamily were undetectable both by immunofluorescence and by RT-PCR analysis. The expression of CD94/NKG2-A appeared to be responsible for the inability of cells to lyse HLA class I+ target cells. Thus, addition of anti-CD94 monoclonal antibodies of IgM isotype resulted in lysis of autologous target cells. The use of 221 cells transfected with different HLA class I alleles as target cells confirmed the broad class I specificity of CD94/NKG2-A receptor. Our experiments indicate that IL-15 provides an appropriate stimulus to the expression of CD94/NKG2-A, but not of other class I-specific NK receptors in the process of maturation of NK cells from thymocyte precursors.

Alleles↗

Inhibitory receptors for HLA class I molecules on cytolytic T lymphocytes. Functional relevance and implications for anti-tumor immune responses.

In recent years, the molecular mechanism by which natural killer cells lyze, or fail to lyze, different target cells has been elucidated. Natural killer cells express receptors which recognize MHC class I molecules on target cells. This interaction leads to inhibition of cytolytic activity, thus preventing lysis of target cells. The receptors belong to two distinct molecular types: (1) the lg superfamily which includes receptors (p58.1, p58.2, p70, and p140) which recognize specific HLA allotypes; (2) CD94 molecules which display a broad specificity for HLA class I molecules. Recently, a subset of cytolytic T lymphocytes has been shown to express the various natural killer cell receptors. Such T cells are detectable in peripheral blood, spleen, tonsils, and lymph nodes, but not in the thymus and cord blood. In some instances, two or more natural killer receptors can be coexpressed at the single cell level. Surface marker analysis has revealed that natural killer cell receptor-positive T cells always express a memory phenotype. Moreover, they are characterized by a skewed T cell receptor V beta repertoire. Further analysis of the T cell receptor VDI sequences revealed that natural killer cell receptor-positive, CD3-positive cells isolated from a given individual are oligoclonal or monoclonal in nature. Crosslinking of natural killer receptors leads to inhibition of different T cell functions, including non-specific lysis of appropriate HLA class I-negative target cells, T cell receptor mediated cytotoxicity, and cytokine production. The inhibitory effect on T cell receptor-mediated function has important implications. Thus, the expression of natural killer cell receptors as a consequence of chronic antigen stimulation may result in functional impairment of specific cytolytic T lymphocytes. Preliminary data indicate that this phenomenon may occur in tumor or virally infected patients. Remarkably, various patients with large granular lymphocyte expansions characterized by a CD3/ natural killer receptor-positive phenotype had chronic viral infections. The fact that antigen-specific cytolytic T lymphocytes may simultaneously express T cell and natural killer cell receptors, both recognizing HLA class I molecules but mediating opposite signals, offers new perspectives in our appreciation of the regulation of T cell responses and offers new clues for understanding the immunopathological events involved in certain diseases.

Antigens, Neoplasm↗

HLA-class I-specific inhibitory receptors in human cytolytic T lymphocytes: molecular characterization, distribution in lymphoid tissues and co-expression by individual T cells.

A subset of cytolytic T lymphocytes has been shown to express receptors of the NK type (NKR) which can inhibit T cell cytotoxicity induced via the TCR-CD3 pathway. In this study, by the analysis of full length cDNA amplified from representative T cell clones, we show that NKR belonging either to the lg superfamily, including p58.1, p58.2, p70 and p140, or to the C-type lectin superfamily (CD94/NKG2A), display sequences which are identical to those of the corresponding NKR expressed by CD3-NK cells. Moreover, a fragment of cDNA encoding the NKG2A protein was consistently amplified from all CD94+ T cell clones analyzed. Since different NKR types can be expressed by T cells, we analyzed whether individual T cells could co-express more than one NKR. Analysis of either resting or activated (and cultured) T cell populations revealed that two or more NKR can be co-expressed by single T cells. Moreover, by the analysis of T cell clones, we show that co-expressed receptors are functional and can inhibit independently the TCR-induced cytolytic function. Finally, we investigated whether NKR+ T lymphocytes were also present in lymphoid tissues. No such cells were found in thymus or cord blood, thus further supporting the notion that they represent memory T cells. On the other hand, they were present in all the peripheral tissues analyzed including spleen, lymph nodes and tonsils.

Animals↗

Artificial neural network identification of heterotrophic marine bacteria based on their fatty-acid composition.

The traditional approach to biochemical identification of marine fresh isolates requires considerably long culture preparation times and large quantities of expensive materials and reagents, and the results are not reliable. On the other hand, taxonomy tests based on DNA composition, although sensitive and reliable, require long execution time and high costs. A method is presented for the classification of fatty-acid profiles, extracted from marine bacteria strains, at genus level based on supervised artificial neural networks. The proposed method allows the correct identification of all patterns belonging to the test set. Moreover, a quantitative measure of the importance of each fatty acid for bacterial classification is also achieved. This measure allows the determination of a cluster of fatty acids to be controlled with greater care. The results show that the proposed method is reproducible and rapid, so that it can be routinely used in the marine microbiology laboratory to identify fresh isolates.

Algorithms↗

Expression of HLA class I-specific inhibitory receptors by chronically activated cytolytic T lymphocytes: useful or harmful to the host?

T cells, primarily chronically activated cytolytic T lymphocytes (CTL), can express human leukocyte antigen class I-specific inhibitory receptors previously identified in natural killer cells. Engagement of these receptors leads to inhibition of CTL function. Although the receptor expression may represent a safety device to prevent autoimmunity, it may result in the inability to control viral spread or tumour growth, i.e. two main functions of CTLs. The physiopathologic implications of this event are discussed.

Animals↗

Attachment of Vibrio alginolyticus to chitin mediated by chitin-binding proteins.

Vibrio alginolyticus is the only culturable vibrio associated with the chitinaceous carapace of the copepod Tigriopus fulvus (Fisher 1860) living in Ligurian coastal rock pools (Tyrrhenian Sea). The characteristics of the interaction between chitin particles and V. alginolyticus were studied by analysing strains isolated both from the copepod surface and from rock-pool water. The highest degree of attachment to chitin was observed at 20 degrees C, in the presence of 3% NaCl. Bacterial treatment with N-acetylglucosamine and pronase E caused a reduction in attachment of 52-62% and 77-94%, respectively. Chitin pretreatment with either wheat germ agglutinin or membrane proteins (MPs) from V. alginolyticus caused a reduction in attachment, of 50-57% and 53-70%, respectively. No inhibition was observed when bacteria were pretreated with D-glucose, D-fucose or D-fructose, or when chitin was pretreated with concanavalin A and Escherichia coli DH5 alpha MPs. V. alginolyticus MPs able to bind chitin were isolated and analysed by SDS-PAGE. Four chitin-binding proteins were visualized in all tested strains (53, 35, 20 and 14 kDa); in vivo these peptides may efficiently mediate V. alginolyticus attachment to chitin-containing substrates.

Acetylglucosamine↗

Automated systems for identification of heterotrophic marine bacteria on the basis of their Fatty Acid composition.

The fatty acid methyl ester composition of a total of 71 marine strains representing the genera Alteromonas, Deleya, Oceanospirillum, and Vibrio was determined by gas-liquid chromatographic analysis. Over 70 different fatty acids were found. The predominant fatty acids were 16:0, 16:1 cis 9, summed-in-feature (SIF) 4 (15:0 iso 2OH and/or 16:1 trans 9) and SIF 7 (18:1 cis 11, 18:1 trans 9, and/or 18:1 trans 6) for all the strains considered, but minor quantitative variations could be used to distinguish the different genera. In addition to a conventional statistical processing method to analyze the data and draw comparison between species and genera, an approach involving neutral network-based elaboration is applied. The statistical analysis and dendrogram representation gave a comparison of the species considered, while the neural network computation provided a more accurate assignment of species to their genera. Moreover, by using neural networks, it was possible to conclude that only 22 fatty acids were important for the identification of the marine genera considered. A database of Alteromonas, Deleya, Oceanospirillum, and Vibrio fatty acid methyl ester profiles was generated and is now routinely used to identify fresh marine isolates.

Journal Article↗

Occurrence of Vibrio alginolyticus in Ligurian Coast Rock Pools (Tyrrhenian Sea, Italy) and Its Association with the Copepod Tigriopus fulvus (Fisher 1860).

A study of heterotrophic bacteria and vibrios adhering to the copepod Tigriopus fulvus, which lives in Ligurian coast rock pools (Tyrrhenian Sea), was carried out from November 1990 to October 1991. Heterotrophic bacteria, which were always found both free in the water and bound to the T. fulvus organisms, showed a correlation with water temperature and salinity. Vibrio alginolyticus was found free in the water and bound to T. fulvus surfaces during the warmest months. Temperature is the main factor influencing the presence of V. alginolyticus in the rock pool. Attachment of this microorganism to the copepod provides a mechanism for its extended geographic distribution.

Journal Article↗

Interpretation of gascromatographic data via artificial neural networks for the classification of marine bacteria.

We propose a new method for classification of marine bacteria. This method uses gaschromatograms, which contain information of fatty acid percentage contents of the fresh isolate. For the interpretation of these gaschromatograms we use a surpervisioned artificial neural network. We present a preliminary study on this matter, whose first results show good convergence and classification features.

Algorithms↗