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74. Advanced techniques for in situ analysis of the biofilm matrix (structure, composition, dynamics) by means of laser scanning microscopy

Author

Donelli, G., Neu, Thomas, Lawrence, J.R., Donelli, G., Neu, Thomas, and Lawrence, J.R.

Abstract

The extracellular constituents in bioaggregates and biofi lms can be imaged four dimensionally by usinglaser scanning microscopy. In this protocol we provide guidance on how to examine the various extracellularcompartments in between microbial cells and communities associated with interfaces. The currentoptions for fl uorescence staining of matrix compounds and extracellular microhabitats are presented.Furthermore, practical aspects are discussed and useful notes are added. The chapter ends with a briefintroduction to other approaches for EPS analysis and an outlook for future needs.

Published
2014

75. Chip calorimetry for evaluation of biofilm treatment with biocides, antibiotics, and biological agents

Author

Donelli, G., Morais, Frida, Buchholz, Friederike, Maskow, Thomas, Donelli, G., Morais, Frida, Buchholz, Friederike, and Maskow, Thomas

Abstract

Any growth or bioconversion in biofilms is accompanied by the release of heat. The heat (in J) is tightly related to the stoichiometry of the respective process via law of Hess, and the heat production rate (in W or J/s) is additionally related to the process kinetics. This heat and the heat production rate can nowadays be measured by modern calorimetry with extremely high sensitivity. Flow-through calorimetry allows the measurement of bioprocesses in biofilms in real time, without the need of invasive sample preparation and disturbing of biofilm processes. Furthermore, it can be applied for long-term measurements and is even applicable to turbid media. Chip or miniaturized calorimeters have the additional advantages of extremely short thermal equilibration times and the requirement of very small amounts of media and chemicals. The precision of flow-through chip calorimeters (about 3 mW/L) allows the detection of early stages of biofilm development (about 105 bacteriacm−2).

Published
2014

90. Intracellular expression of P-170 glycoprotein in peripheral blood mononuclear cell subsets from healthy donors and HIV-infected patients

Author

walter malorni, Mb, Lucia, Rainaldi G, Cauda R, Cianfriglia M, Donelli G, and Ortona L

Subjects
Adult, Male, PBMC, HIV, HIV Infections, Settore MED/17 - MALATTIE INFETTIVE, Immunophenotyping, Microscopy, Fluorescence, p-170 glycoprotein, HIV-1, Leukocytes, Mononuclear, Humans, Female, ATP Binding Cassette Transporter, Subfamily B, Member 1
Abstract

P-glycoprotein (P-gp) is a transmembrane efflux pump that actively extrude a variety of unrelated drugs from cancer cells, leading to the so-called multidrug resistance (MDR) phenomenon. However, P-gp has also been found in normal bone marrow and peripheral blood mononuclear cells (PBMC). Recently, the presence of P-glycoprotein in PBMC from human immunodeficiency virus (HIV)-infected patients has also been investigated and a phenotype-associated P-gp expression has been detected.A total of thirty-eight HIV-1 positive patients with a mean age of 34 years (range, 24-41 years) were studied after an informed consent. Peripheral blood mononuclear cells (PBMC) were isolated by centrifugation on a Ficoll/Hypaque and P-glycoprotein expression was investigated on lymphocyte population by single and double-color immunofluorescence techniques. We investigated: i) both surface and intracellular expression of the P-gp molecule in different PBMC subsets, ii) P-gp expression modifications occurring during HIV infection, and iii) the effect of HIV-gp120 on the expression of P-gp by T lymphocyte subsets from healthy donors.Our experimental findings indicate that: a) P-gp glycoprotein can be detected on an intracellular level in different PBMC subpopulations (mainly CD8+ T lymphocytes, CD16+ NK cells and CD14+ monocytes); b) this intracellular expression is decreased in specific PBMC subsets (i.e. T-CD8+ and NK-CD16+) from HIV-infected patients and c) a rearrangement was obtained when CD4+ and CD8+ lymphocytes from healthy donors were exposed in vitro to the HIV-binding glycoprotein gp120.Our results indicate that P-gp glycoprotein can also be expressed intracellularly and can be rearranged in PBMC subsets from HIV-infected patients.

Published
1998

95. Induction of phagocytic behaviour in human epithelial cells by Escherichia coli cytotoxic necroziting factor type 1

Author

Falzano, L., Fiorentini, C., Donelli, G., Michel, E., Kocks, C., Cossart, Pascale, Cabanié, L., Oswald, E., Boquet, P., Microbiologie, Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB), and ProdInra, Migration

Subjects
POUVOIR PATHOGENE, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology
Published
1993

97. A new, striking morphological alteration of P-glycoprotein expression in NK cells from AIDS patients

Author

Cauda, Roberto, Ortona, Luigi, Lucia, Mothanje Barbara Patricia, Rainaldi, G, Donelli, G, Malorni, W., Cauda, Roberto (ORCID:0000-0002-1498-4229), Lucia, Mothanje Barbara Patricia (ORCID:0000-0001-7648-2198), Cauda, Roberto, Ortona, Luigi, Lucia, Mothanje Barbara Patricia, Rainaldi, G, Donelli, G, Malorni, W., Cauda, Roberto (ORCID:0000-0002-1498-4229), and Lucia, Mothanje Barbara Patricia (ORCID:0000-0001-7648-2198)

Abstract

Natural killer cells from healthy donors express P-glycoprotein on their surface. This molecule is rearranged during the process of cell-mediated cytotoxicity and it appears to be clustered in the cell-to-cell contact regions. By contrast, in HIV-infected subjects this rearrangement is hindered. These results seem to be associated with cytoskeleton network alterations of the cell-mediated killing process occurring in AIDS patients and can contribute to the comprehension of the mechanisms of the natural killer cell deficiency found in these patients.

Published
2000

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