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In this paper, a new method is described for the horizontal electrophoresis of cells on a density cushion under near-isopycnic conditions. When cell sedimentation is minimized, the electrophoresis of red blood cells (RBC) used as model cells within an anti-convective porous matrix (with pores over 300 μm in diameter) was capable of separating a mixture of human and chicken RBC according to their electrophoretic mobilities. Samples taken from the separated RBC bands show over 90% purity for each species. The simultaneous electrophoresis of several RBC samples carried out under identical conditions permitted the use of comparative data based on the electrophoretic mobility of cells which differ in their surface properties. We believe that this relatively simple system, in which cell sedimentation and convection are minimized, has the potential to be modified and adapted for the separation of other cell types/organelles.
A bridged peptide with the sequence: H-Thr-Pro-Gln-Arg-Gly-Asp-Val-y-Abu-Asn- Asp-Gln-Glu-Glu-Thr-Thr-Gly-Val-Val-Ser-Thr-Pro-Leu-Ile-Arg-Asn-Gly-OH was de­signed to mimic the discontinuous epitope of the HLA-DQ molecule that might interact with CD4. The bridged peptide revealed distinct suppressory effect in the humoral im­mune response. This result supports our suggestion that the 164-172 region of the HLA-DQ molecule may enhance its interactions with coreceptors, possibly with CD4.
LacCer/CDw17 is the most abundant GSL in neutrophils. The cell-surface and intracellular presence of LacCer was determined quantitatively using anti-CDw17 mAbs in a flow cytometry assay. The quantified alterations in the level of CDw17 antigen expression are consistent with alterations in LacCer content, determined chemically. Our results show that CDw17 antigen expression defines successive stages in the maturation of the myeloid cell. The assessment of cell-surface and intracellular CDw17 expression may be useful in evaluating neutrophil physiological status.
The bacterial strains from the genera: Bacillus, Pseudomonas, Aeromonas, Achromobacter, and Flavimonas isolated from soil contaminated with crude oil were the subject of studies. The effect of the addition of rhamnolipids on cell surface properties and the removal efficiency of diesel oil were investigated. Rhamnolipids caused the modification of cell surface properties of tested strains, which depended on the amount of external additions of biosurfactant. Additionally, the decrease of Zeta potential was observed after the introduction of rhamnolipids to the diesel oil system. Particle size distribution provides information about system homogeneity and the tendency of particles toward agglomeration. Cell surface hydrophobicity during hydrocarbon biodegradation is a dynamic parameter. There were no different effects, after the addition of rhamnolipids, on the Gram positive and Gram negative bacterial strains. Moreover, the addition of rhamnolipids does not always increase the efficiency of diesel oil biodegradation. The most effective bacterial strain in diesel oil removal was the genus Pseudomonas.
This report describes an improvement made to the horizontal cell electrophoresis methodology. It involves using two liquid layers differing in density to produce an interface described as a “density cushion”. The electrophoretic system that employed an anti-convective porous matrix to separate red blood cells (RBC) and charged dyes effectively was found to be unsuitable for some other mammalian cells. The “density cushion” method was found to be more versatile and applicable to studies on the separation of a variety of cell types. The experiments described show the differences between the electrophoretic mobilities of a human eosinophilic leukaemia cell line (Eol-1) and RBC, both with and without the modification of the cell surface properties.
The collection of 314 staphylococcal strains including Staphylococcus aureus and coagulase-negative staphylococci (CNS) was isolated from skin or nasopharynx of healthy people. It was found that the majority of staphylococci possessed the ability to produce slime intensively or moderately, irrespective of ecological niche-nose, throat or skin. Most of them showed the hydrophilic cell surface. However, among S. aureus skin isolates or CNS throat isolates predominated strains with hydrophobic cell surface. There was a slight correlation between slime production and the nature of cell surface among CNS isolates but not among S. aureus strains. It was found that most of slime-producing CNS strains showed hydrophilic cell surface, while slime-negative isolates usually possessed hydrophobic cell surface. Our data suggest that slime production but not cell surface hydrophobicity can be regarded as an essential colonization factor responsible for staphylococci adherence to skin or mucous membranes of upper respiratory tract. These data also suggest that slime production seems to be a general feature of staphylococci isolated from various niches of healthy people.
Culture of Papaver somniferum in vitro was used for a characterisation of cell surface structures and mode of cell adhesion and cell separation during cell differentiation and plant regeneration in somatic embryogenesis and shoot organogenesis. In early stages of somatic embryogenesis, cell type-specific and developmentally regulated change of cell morphogenesis was demonstrated. Cell wall of separated embryonic cells were self-covered with external tubular network, whereas morphogenetic co-ordination of adhered cells of somatic proembryos was supported by fine and fibrillar external cell wall continuum of peripheral cells, interconnecting also local sites of cell separation. Such type of cell contacts disappeared during histogenesis, when the protodermis formation took place. Tight cell adhesion of activated cells with polar cell wall thickening, and production of extent mucilage on the periphery were the crucial aspects of meristemoids. Fine amorphous layer covered developing shoot primordia, but we have not observed such comparable external fibrillar network. On the contrary intercellular separation of differentiated cells in regenerated organs, and accepting distinct developmental system of somatic embryogenesis and shoot organogenesis, cell adhesion in early stages and ultrastructural changes associated with tissue disorganisation, and the subsequent reorganisation into either embryos or shoots appear to be regulatory morphogenetical events of plant regeneration in vitro.
Fas ligand (L) is a membrane protein from the tumor necrosis factor (TNF) family. It induces apoptosis upon contact with its Fas/CD95/APO1 receptor. Trimerization of FasL on the surface of effector cells is essential in the binding of the Fas trimer of the target cells. The receptor then recruits an adaptor and caspase-like proteins which lead apoptosis. This paper reports on the fate of FasL in HEp-2 cells committed to apoptosis by induction with campthotecin. Our main results demonstrated that in non-apoptotic cells, FasL aggregates in the cytoplasm forming trimers of 120 kDa. Apoptosis increases the trimeric FasL species, but also induces its dissociation into monomers of 35 kDa. In conclusion, camptothecin appears to perturb the Fas and FasL segregation in the cytoplasm by promoting the transit of FasL to the cell surface, thus fostering a process of autocrine or paracrine apoptosis. FasL is trimerized prior to Fas/FasL complex formation, and after apoptosis, FasL undergoes an intense turnover.
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