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By using the lH—NMR method and paramagnetic ions Pr3+, the incorporation into the liposome membrane of a series of biologically active single-chain (TGE-n) and double-chain (DGE-n) quaternary ammonium salts was studied. Corresponding compounds of both groups differed in their alkyl chain-length (n = 8,10,12,14 and 16). Minimal concentration CA of compounds TGE-n and DGE-n which caused total release of Pr3+ ions from the external layer of liposome membrane and the ratio of CA to the 4.5 mM extravesicular concentration of Pr3+ (CPr) used in experiments was determined. Based on the results obtained, one can conclude that the effectiveness of Pr3+ release from liposome membrane is (with n = 12,14 and 16) twice as high for double-chain compounds as for analogous single-chain compounds, and about 2.5 and 5 times higher for DGE-10 and DGE-8 than for TGE-10 and TGE-8, respectively.
The degree of dependence of a lipid bilayer’s surface properties on its conformational state is still an unresolved question. Surface properties are functions of molecular organization in the complex interfacial region. In the past, they were frequently measured using fluorescence spectroscopy. Since a fluorescent probe provides information on its local environment, there is a need to estimate the effect caused by the probe itself. In this paper, we address this question by calculating how lipid head-group orientation effects the fluorescence intensity of Fluorescein-PE (a probe that is sensitive to surface potential). In the theoretical model assumed the lipid bilayer state and the interactions between the charged fluorescent probe and the surrounding lipid molecules was evaluated. The results of this theoretical analysis were compared with experimentally obtained data. A lipid bilayer formed from DPPC was chosen as the experimental system, since it exhibits all the major conformational states within a narrow temperature range of 30°C - 45°C. Fluorescein-PE fluorescence intensity depends on local pH, which in turn is sensitive to local electrostatic potential in the probe’s vicinity. This local electrostatic potential is generated by lipid head-group dipole orientation. We have shown that the effect of the probe on lipid bilayer properties is limited when the lipid bilayer is in the gel phase, whereas it is more pronounced when the membrane is liquid-crystalline. This implies that Fluorescein-PE is a good reporter of local electrostatic fields when the lipid bilayer is in the gel phase, and is a poor reporter when the membrane is in the liquid-crystalline state.
The constant-current chronopotentiometric measurements of egg yolk phosphatidylcholine bilayer membrane without and with cholesterol are presented. It was demonstrated that the constant-intensity current flow through the bilayer membranes without and with cholesterol generated the oscillating pores in their structures. The presence of cholesterol in the bilayer membrane increased the value of critical potential at which pores could be formed. The shift in the distribution of calculated pore radii towards smaller values was observed in the bilayer containing cholesterol. It was postulated that greater stability of bilayer with cholesterol resulted from increased critical pore radius (at which the bilayer would rupture). The implications of the membrane cholesterol for the application of constant-current method as a biotechnological tool for incorporating molecules of different size are discussed.
Cereal grain resorcinolic lipids (S-n-alk(en)ylresorcinols) at micromolar concentrations are able to protect the erythrocyte membrane against hydrogen peroxide-induced lipid oxidation. The antioxidative effect is dependent upon chain length of alkylresorcinol molecules. The C15:0 homolog (IC50 of 10 uM) exhibited strongest activity whereas for long chain homologs (C19: 0 and C23 : 0) IC50 values were higher, 32.5 and 59 uM, respectively. The protective effect of alkylresorcinolic antioxidants was also dependent on their incorporation into the membrane, that is governed by their water-membrane partition coefficient The results obtained show that alkylresorcinols should be recognized as hydrophobic, membrane-localised antioxidants.
Spectral properties (anisotropy coefficients calculated for absorption, emission and fluorescence decay time) of two stilbazolium merocyanine dyes have been determined to evaluate the applicability of these dyes as sensitizers in photodynamic therapy. The dyes were embedded in an anisotropic polymer matrix. Analysis of the emission decay components measured in polarized light provides information on the interactions of the dye molecules with the polymer matrix being a model of an anisotropic biological system. Different values of the emission anisotropies obtained from various polarized components of fluorescence decays have shown that the orientations of the dye molecules influence their interactions with the polymer. This means that differently oriented dye molecules located in biological systems should exhibit different interactions with membranes. The chain length and type of side groups attached as well as the salt form of the dye molecule were shown to influence the dye-polymer interactions and should be taken into account before the application of merocyanine dyes in medicine. These dyes seem to be promising optical sensors with spectral properties, including the calculated anisotropy coefficients, sensitive to the molecular environment, useful to study orientation and interaction with neighbouring molecules in biological membranes.
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