Search results for "erythrocyte membrane"

Red cell Ca2+ content (total and cytosolic) and erythrocyte membrane fluidity in several clinical conditions

1993

Techniques to evaluate erythrocyte deformability in diabetes mellitus

2005

Using several rheological techniques, we examined the erythrocyte deformability in different groups of diabetic subjects. The macrorheological techniques used for this evaluation were respectively whole-blood filtration, filtration of erythrocyte suspensions, polyviscosimetry and diffractometry. Whole-blood filterability, at a negative pressure of 20 cm water, was decreased in type 2 diabetics; no difference was evident at a negative pressure of 10 cm water. The filtration of erythrocyte suspensions at low haematocrit (5%) did not show differences between normal and diabetic subjects. The polyviscosimetry, which explores the filterability of erythrocyte suspensions at high haematocrit (80%)…

Eventi vascolari correlati ad alcune condizioni di iperviscosità sclerocitemica

2014

Altered pore-forming properties of proteolytically nicked staphylococcal alpha-toxin

1993

Staphylococcal alpha-toxin is a single-chain polypeptide with a molecular weight of 34,000 that hexamerizes in lipid bilayers to form pores of 1-1.5 nm effective diameter in membranes. We demonstrate that limited proteolysis of purified alpha-toxin with proteinase K generates a hemolytically active product that yields one major protein band of 17-18 kDa in SDS-polyacrylamide gel electrophoresis. The 17-18-kDa protein band harbors two major fragments of similar size representing the N- and C-terminal halves, which remain associated with each other in non-denaturing buffers but dissociate in 6 M urea. Dissociation in urea leads to loss of hemolytic activity. In contrast, unnicked alpha-toxin …

Staphylococcal alpha-toxin: formation of the heptameric pore is partially cooperative and proceeds through multiple intermediate stages.

1997

Staphylococcal alpha-toxin is a 293 residue polypeptide that assembles into pore-forming heptamers, residues 118-140, thereby inserting to form an amphipathic beta-barrel in the lipid bilayer. Fluorometric analyses were here conducted using cysteine-substitution mutants site-specifically-labeled at positions 35 or 130 with the environmentally-sensitive fluorophore acrylodan. In conjunction with functional assays, three conformational states of the heptamer were defined, which may represent transitional configurations of the toxin molecule along its way to membrane insertion and pore formation. The first was the freshly assembled, SDS-sensitive heptamer alpha7*a, where a minor alteration in …

The hemagglutinin of Staphylococcus saprophyticus is a major adhesin for uroepithelial cells.

1996

The 160-kDa hemagglutinin of Staphylococcus saprophyticus also serves as a fibronectin-binding protein, and the two activities may be present on different parts of the molecule. Bacteria expressing the 160-kDa hemagglutinin bound in large numbers to histological sections of human ureters, whereas nonhemagglutinating bacteria did not bind. Binding was decreased by an antiserum to the 160-kDa protein and by a preparation of sheep erythrocyte membranes. Fibronectin had no effect. We therefore conclude that binding of S. saprophyticus to uroepithelial cells is mediated by the hemagglutinating activity of the 160-kDa surface protein.

Putative identification of an amphipathic alpha-helical sequence in hemolysin of Escherichia coli (HlyA) involved in transmembrane pore formation.

2008

Abstract Escherichia coli hemolysin is a pore-forming protein belonging to the RTX toxin family. Cysteine scanning mutagenesis was performed to characterize the putative pore-forming domain of the molecule. A single cysteine residue was introduced at 48 positions within the sequence spanning residues 170–400 and labeled with the polarity-sensitive dye badan. Spectrofluorimetric analyses indicated that several amino acids in this domain are inserted into the lipid bilayer during pore formation. An amphipathic α-helix spanning residues 272–298 was identified that may line the aqueous pore. The importance of this sequence was highlighted by the introduction of two prolines at positions 284 and…

Pyrrolo[2,3-h]quinolinones: A new ring system with potent photoantiproliferative activity

2006

A new class of compounds, the pyrrolo[2,3-h]quinolin-2-ones, nitrogen isosters of the angular furocoumarin Angelicin, was synthesized with the aim of obtaining new photochemotherapeutic agents with increased antiproliferative activity and lower undesired toxic effects than the lead compound. Two synthetic pathways were approached to allow the isolation both of the dihydroderivatives 10-17 and of the aromatic ring system 23. Compounds 10-17 showed a remarkable phototoxicity and a great UVA dose dependence reaching IC(50) values at submicromolar level. Intracellular localization of these compounds has been evaluated by means of fluorescence microscopy using tetramethylrhodamine methyl ester a…

α-Tocopherol Modulates Phosphatidylserine Externalization in Erythrocytes

2006

Objective— The aim of the present study was to assess the effect of α-tocopherol, the main vitamin E isomer on phosphatidylserine (PS) exposure at the surface of circulating erythrocytes, and to determine consequences on erythrocyte properties. Methods and Results— In vitro α-tocopherol enrichment of isolated erythrocytes significantly decreased PS externalization as assessed by lower Annexin V-fluorescein isothiocyanate labeling. Plasma phospholipid transfer protein (PLTP) transfers vitamin E, and both α-and γ-tocopherol accumulated in circulating erythrocytes from PLTP-deficient homozygous (PLTP −/− ) mice as compared with wild-type mice. In agreement with in vitro studies, vitamin E–enr…

Diabetes mellitus: evaluation of erythrocyte and polymorphonuclear leukocyte rheology

2015

Aim: To explore red blood cells (RBC) and leukocyte rheology, that may be relevant in the pathophysiology of diabetes mellitus. Methods and Results: Significant alterations have been observed in RBC behaviour using several filtration techniques, but the exploration of RBC deformability by laser diffractometry did not show any abnormality. We have also employed microrheological methods based on fluorescence spectroscopy: membrane microviscosity was evaluated in ghosts, while in intact RBC we explored the membrane polarity gradient using fluorescent fatty acids, the phospholipid and protein lateral mobility using respectively pyrene and pyrene-3-maleimide. Alterations emerged only using the l…