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See detailCell Handling, Membrane-Binding Properties, And Membrane-Penetration Modeling Approaches Of Pivampicillin And Phthalimidomethylampicillin, Two Basic Esters Of Ampicillin, In Comparison With Chloroquine And Azithromycin
Chanteux, H.; Paternotte, I.; Mingeot-Leclercq, Mp. et al

in Pharmaceutical Research (2003), 20(4), 624-31

PURPOSE: The purpose of this work was to examine and understand the cellular pharmacokinetics of two basic esters of ampicillin, pivaloyloxymethyl (PIVA) and phthalimidomethyl (PIMA), in comparison with ... [more ▼]

PURPOSE: The purpose of this work was to examine and understand the cellular pharmacokinetics of two basic esters of ampicillin, pivaloyloxymethyl (PIVA) and phthalimidomethyl (PIMA), in comparison with lysosomotropic drugs (chloroquine, azithromycin). METHODS: Cell culture studies (J774 macrophages) were undertaken to study uptake and release kinetics and to assess the influence of concentration, pH, proton ionophore (monensin), and MRP and P-gp inhibitors (probenecid, gemfibrozil, cyclosporin A, GF 120918). Equilibrium dialysis with liposomes were performed to directly asses the extent of drug binding to bilayers. Conformational analysis modeling of the drug penetration in bilayers was conducted to rationalize the experimental observations. RESULTS: PIVA and PIMA showed properties in almost complete contrast with those of chloroquine and azithromycin, i.e., fast apparent accumulation and fast release at 4 degrees C as well as at 37 degrees C, saturation of uptake (apparent Kd 40 microM), no influence of monensin, MRP, or P-gp inhibitors; tight binding to liposomes (Kd approx. 40 microM); and sharp increase in calculated free energy when forced in the hydrophobic domain. CONCLUSIONS: Although they are weak organic bases, PIVA and PIMA show none of the properties of lysosomotropic agents. We hypothesize that they remain locked onto the pericellular membrane and may never penetrate cells as such in significant amounts. [less ▲]

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See detailCell invasion in the spheroid sprouting assay: a spatial organisation analysis adaptable to cell behaviour.
Blacher, Silvia ULg; Erpicum, Charlotte ULg; Lenoir, Benedicte et al

in PLoS ONE (2014), 9(5), 97019

The endothelial cell spheroid assay provides a suitable in vitro model to study (lymph) angiogenesis and test pro- and anti-(lymph) angiogenic factors or drugs. Usually, the extent of cell invasion ... [more ▼]

The endothelial cell spheroid assay provides a suitable in vitro model to study (lymph) angiogenesis and test pro- and anti-(lymph) angiogenic factors or drugs. Usually, the extent of cell invasion, observed through optical microscopy, is measured. The present study proposes the spatial distribution of migrated cells as a new descriptor of the (lymph) angiogenic response. The utility of this novel method rests with its capacity to locally characterise spheroid structure, allowing not only the investigation of single and collective cell invasion but also the evolution of the spheroid core itself. Moreover, the proposed method can be applied to 2D-projected spheroid images obtained by optical microscopy, as well as to 3D images acquired by confocal microscopy. To validate the proposed methodology, endothelial cell invasion was evaluated under different experimental conditions. The results were compared with widely used global parameters. The comparison shows that our method prevents local spheroid modifications from being overlooked and leading to the possible misinterpretation of results. [less ▲]

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See detailCell localization and redistribution of the 67 kD laminin receptor and alpha 6 beta 1 integrin subunits in response to laminin stimulation: an immunogold electron microscopy study.
Romanov, V.; Sobel, M. E.; pinto da Silva, P. et al

in Cell Adhesion and Communication (1994), 2(3), 201-9

The 67 kDa laminin receptor (67LR), one of several cell surface laminin-binding proteins, is involved in the interactions between cancer cells and laminin during tumor invasion and metastasis. A 37 kDa ... [more ▼]

The 67 kDa laminin receptor (67LR), one of several cell surface laminin-binding proteins, is involved in the interactions between cancer cells and laminin during tumor invasion and metastasis. A 37 kDa polypeptide (37LRP), previously identified as the 67LR precursor, is abundantly present in the cytoplasm and has been implicated in polysome formation. To better understand the cellular localization of the 67LR and its precursor, transmission electron microscopic studies of human melanoma A2058 cells were carried out using immunogold labeling and a variety of antibodies: (a) affinity purified antibodies directed against 37LRP cDNA-derived synthetic peptides; (b) anti-67LR monoclonal antibodies raised against intact human small cell lung carcinoma cells; and (c) monoclonal antibodies against the subunits of the integrin laminin receptor, alpha 6 beta 1. Double-labeling immunocyto-chemistry revealed that anti-67LR monoclonal antibodies as well as anti-37 LRP antibodies recognized antigens that were localized in the cytoplasm in electron dense structures. As expected, cell membrane labeling was also observed. Surprisingly, alpha 6 and beta 1 integrin subunits were detected in the same cytoplasmic structures positive for the 67LR and the 37LRP. After addition of soluble laminin to A2058 cells in suspension, the number of labeled cytoplasmic structures increased especially in the vicinity of the plasma membrane, and were exported onto the cell surface. Neither fibronectin nor BSA induced such an effect. The data demonstrate that the 67 LR and the 37 LRP antibodies detect colocalized antigens that are in cytoplasmic structures with alpha 6 beta 1 integrin.(ABSTRACT TRUNCATED AT 250 WORDS) [less ▲]

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See detailCell lysis during coccolithophorid blooms in the Northern Bay of Biscay
De Bodt, Caroline; Harlay, Jérôme ULg; Roevros, Nathalie et al

Poster (2009, January 25)

Phytoplankton cell lysis occurs in natural populations and is often associated with viral activity and zooplankton grazing. Cell lysis rates are expected to increase towards the decaying phase of the ... [more ▼]

Phytoplankton cell lysis occurs in natural populations and is often associated with viral activity and zooplankton grazing. Cell lysis rates are expected to increase towards the decaying phase of the bloom and may be associated with enhanced microbial activity and export of particulate matter to the seafloor. Their estimation was based on the measurement of esterase (a cytoplasmic enzyme) activity expected to appear in the water only after cell breakage. Field investigations, supported by remote sensing data, were conducted in recent years during late spring in the Northern Bay of Biscay, where frequent and recurrent coccolithophorid blooms are observed. Results on cell lysis rates determined in surface waters will be presented with relevant biogeochemical parameters (temperature, particulate organic and inorganic carbon, transparent exopolymer particles, nutrients, chlorophyll a) in order to investigate phytoplankton dynamics in relation to coccolithophorid development. The use of this parameter to characterize bloom termination, especially during coccolithophorid blooms will be discussed. [less ▲]

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See detailCell membrane proteomic analysis identifies proteins differentially expressed in osteotropic human breast cancer cells.
Kischel, Philippe ULg; Guillonneau, Francois; Dumont, Bruno ULg et al

in Neoplasia : An International Journal for Oncology Research (2008), 10(9), 1014-20

Metastatic breast cancer cells are characterized by their high propensity to colonize the skeleton and form bone metastases, causing major morbidity and mortality. Identifying key proteins involved in the ... [more ▼]

Metastatic breast cancer cells are characterized by their high propensity to colonize the skeleton and form bone metastases, causing major morbidity and mortality. Identifying key proteins involved in the osteotropic phenotype would represent a major step toward the development of both new prognostic markers and new effective therapies. Cell surface proteins differentially expressed in cancer cells are preferred potential targets for antibody-based targeted therapies. In this study, using cell surface biotinylation and a mass spectrometric approach, we have compared the profile of accessible cell surface proteins between the human breast cancer cell line MDA-MB-231 and its highly osteotropic B02 subclone. This strategy allowed the identification of several proteins either up- or downregulated in the osteotropic cell line, and differential protein expressions were validated using antibody-based techniques. Class I HLAs were down-regulated in the bone metastatic variant, whereas alpha(v)beta(3) integrins, among others, were consistently up-regulated in this latter cell line. These results show that comprehensive profiling of the cell surface proteome of mother cancerous cell lines and derived organ-specific metastatic cell lines provides an effective approach for the identification of potential accessible marker proteins for both prognosis and antibody-based targeted therapies. [less ▲]

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See detailCell Models Adapted to Real-Time Imaging of the Cytoskeleton Dynamics in Altered Gravity
Willems, Jérôme ULg; Deroanne, Christophe ULg; Colige, Alain ULg et al

in Microgravity Science and Technology (2014), 26(4), 257-270

Spatial and temporal regulation of cell phenotype by mechanical forces is a growing field of research in health sciences since these stimuli influence cellular functions, such as proliferation, migration ... [more ▼]

Spatial and temporal regulation of cell phenotype by mechanical forces is a growing field of research in health sciences since these stimuli influence cellular functions, such as proliferation, migration, differentiation and gene expression. In the context of the Fluolive project selected by the European Space Agency and aiming at evaluating the impact of gravity alterations on the cell phenotype, we have developed new bone-derived cell lines adapted for live-cell imaging of the cytoskeleton. Osteoblastic cells derived from human osteosarcomas were used as experimental models. U2-OS and SaoS-2 cells stably expressing TagGFP2- β-actin and mCherry- α-tubulin were established and single-cell clonal cultures were characterized in terms of recombinant proteins production and localization, fluorescence intensity, cell proliferation and migration rates. Living fluorescently-tagged cell lines allow real-time fluorescence microscopy of the cytoskeleton dynamics without bleaching and without alteration of cell morphology. U2-OS and SaoS-2 TagGFP2- β-actin and mCherry- α-tubulin clones will be used to monitor the effect of mechanical forces in models of altered gravity on Earth and possibly on the ISS. © 2014, Springer Science+Business Media Dordrecht. [less ▲]

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See detailCell proliferation in malignant melanoma: relationship with neoplastic progression.
PIERARD, Gérald ULg

in ISRN Dermatology (2012), 2012(art.828146), 1-12

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See detailThe cell separation step of bacterial cell division
Kerff, Frédéric ULg

Scientific conference (2013, November 26)

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See detailCell specific expression of Hsp70 in neurons and glia of the rat hippocampus after hyperthermia and kainic acid-induced seizure activity.
Krueger, A. M.; Armstrong, J. N.; Plumier, Jean-Christophe ULg et al

in Molecular Brain Research (1999), 71(2), 265-78

In this study we investigated the time course, cell-type and stress-specific expression of hsp70 mRNA and Hsp70 protein in glial cells and neurons in the rat brain following heat shock treatment and ... [more ▼]

In this study we investigated the time course, cell-type and stress-specific expression of hsp70 mRNA and Hsp70 protein in glial cells and neurons in the rat brain following heat shock treatment and kainic acid-induced status epilepticus. Transcripts for hsp70 were detected in hippocampal homogenates from 1.5 to 6 h following hyperthermia and from 3 to 24 h following kainic acid-induced seizures. In situ hybridization revealed hsp70 mRNA to be region specific and time-dependent following hyperthermia and kainic acid-induced seizures. Western analysis indicated that Hsp70 reached maximal levels at 3 h after hyperthermia and 12 h after kainic acid-induced seizures. Immunohistochemistry revealed low level expression of Hsp70 protein in dentate granule cells at 1.5 and 3 h after hyperthermia. No Hsp70 protein was detected in neurons of the pyramidal cell layer or dentate hilus at any time following hyperthermia. Small Hsp70-immunoreactive cells were detected throughout the hippocampus following hyperthermia that, based on cell size, distribution, and double-labeling with vimentin, were considered to be glia. In contrast, high levels of Hsp70 protein were detected in neurons of the pyramidal cell layer and dentate hilus at 24 h after seizure-inducing kainic acid injection. These results suggest that expression of Hsp70 protein is cell-specific depending on the stressor. In addition, finding high levels of Hsp70 mRNA in the dentate granule cells after hyperthermia, but little or no Hsp70 protein, suggests that the synthesis of the protein is also regulated at the post-transcriptional level following hyperthermia. [less ▲]

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See detailCell surface binding of TIMP-2 and pro-MMP-2/TIMP-2 complex
Emmert-Buck, M. R.; Emonard, H. P.; Corcoran, M. L. et al

in FEBS Letters (1995), 364(1), 28-32

Tissue inhibitor of metalloproteinases (TIMP-2) is a low molecular weight proteinase inhibitor capable of inhibiting activated matrix metalloproteinases (MMPs). TIMP-2 is found both free and in a 1:1 ... [more ▼]

Tissue inhibitor of metalloproteinases (TIMP-2) is a low molecular weight proteinase inhibitor capable of inhibiting activated matrix metalloproteinases (MMPs). TIMP-2 is found both free and in a 1:1 stoichiometric complex with the pro-enzyme form of MMP-2 (pro-MMP-2/TIMP-2 complex). We have measured the binding of recombinant TIMP-2 to intact HT-1080 and MCF-7 cells. HT-1080 cells in suspension bound 125I-labeled rTIMP-2 with a Kd of 2.5 nM and 30,000 sites/cell. Monolayers of MCF-7 cells were similarly found to bind [125I]rTIMP-2 with a Kd of 1.6 nM and 25,000 sites/cell. Specific binding of MMP-2 alone to HT-1080 cells was not observed; however, pro-MMP-2/TIMP-2 complex was capable of binding to the surface of HT-1080 cells in a TIMP-2-dependent manner. Binding of rTIMP-2 was not competed by the presence of TIMP-1. These results suggest that rTIMP-2 alone binds directly to the cell surface of HT-1080 and MCF-7 cell lines, and TIMP-2 is capable of localizing MMP-2 to the surface of HT-1080 cells via interaction with a specific binding site. [less ▲]

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See detailCell surface immunophenotype and gelatinase activity of the human breast carcinoma cell line (MCF-7/6) with functionally defective E-cadherin.
Hlavcak, P.; Sedlakova, O.; Sedlak, J. et al

in Neoplasma (1999), 46(1), 12-6

Expression of differentiation and adhesion cell surface antigens (LewisX - CD15, CD44, syndecan 1 - CD138 and basigin/EMMPRIN - CD147) were determined on the cell surface of human breast carcinoma MCF7 ... [more ▼]

Expression of differentiation and adhesion cell surface antigens (LewisX - CD15, CD44, syndecan 1 - CD138 and basigin/EMMPRIN - CD147) were determined on the cell surface of human breast carcinoma MCF7 cells in vitro with the aid of flow cytometry and compared with that of MCF-7/6 cells, with functionally defective E-cadherin system and increased biological aggressiveness. The major cell surface alterations in MCF-7/6 cells compared with the parental MCF-7 cell line were a markedly increased CD15 (LewisX) and CD44 antigen cell surface expression on MCF-7/6 cells. There were no major differences between parental MCF-7 and MCF-7/6 cells in cell surface syndecan 1, basigin/EMMPRIN, E-cadherin and high affinity non-integrin laminin receptor expression. The constitutive cell surface gelatinase A and B activities were absent on MCF-7 and faint in MCF-7/6 cells. Both phorbol ester TPA and tumor necrosis factor TNF-alpha induced a marked up-regulation of gelatinase B only in MCF-7/6 cells. No marked differences in penetration of MCF-7 vs. MCF-7/6 cells into collagen/fibroblast matrix in vitro were observed. The increased expression of CD15 (LewisX), CD44 antigen and TNF-alpha-inducible gelatinase B on MCF-7/6 cells may represent auxiliary factors contributing to the increased biological aggressiveness of MCF-7/6 cells. [less ▲]

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See detailCell Surface Receptors in Lymphoid Cells: From Cytochemistry to Molecular Biology and from a Phenotype to a Function
Boniver, Jacques ULg; Courtoy, R.; Schaaf-Lafontaine, Nicole ULg et al

in Progress in Histochemistry and Cytochemistry (1992), 26(1-4), 169-81

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See detailCell survival and preservation of siRNA-mediated protein knock-down upon serum-free cryopreservation (-80 degrees C).
Lambert, Charles ULg; Deroanne, Christophe ULg; Servotte, Sandrine et al

in Gravitational and Space Biology Bulletin (2005), 18(2), 103-4

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See detailCell turnover in BLV-infected sheep
Debacq, Christophe; Peremans, T.; Kerkhofs, Pierre et al

in Aids Research and Human Retroviruses": 10th International Conference on Human Retrovirology: HTLV and Related Viruses, (2001, June)

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