Torocyte Membrane Endovesicles Induced by Octaethyleneglycol Dodecylether in Human Erythrocytes

AbstractEndovesicles induced in human erythrocytes by octaethyleneglycol dodecylether (C12E8) were studied by confocal laser scanning microscopy, using fluorescein isothiocyanate dextran as a nonspecific fluid marker. The endovesicles appeared to consist mainly of a ring-formed toroidal part joined with a central flat membrane segment. The torocyte contour length was several μm. There was usually one torocyte endovesicle per cell. The endovesicles seemed to be located near the cell surface. In sections of C12E8-treated erythrocytes transmission electron microscopy revealed the frequent occurrence of flat membrane structures with a bulby periphery, which apparently are cross sections of torocyte endovesicles. The possible physical mechanisms leading to the observed torocyte endovesicle shape are discussed. The torocyte endovesicles seem to be formed in a process in which an initially stomatocytic invagination loses volume while maintaining a large surface area. Because intercalation of C12E8 in the erythrocyte membrane induces inward membrane bending (stomatocytosis) we assume that C12E8 is preferentially located in the inner lipid layer of the erythrocyte membrane, i.e., in the outer lipid layer of the endovesicle membrane. It is suggested that local disturbances of the lipid molecules in the vicinity of the C12E8 molecules in the outer lipid layer of the endovesicle membrane form membrane inclusions with the effective shape of an inverted truncated cone. If the interaction between the inclusion and the membrane is weak, the membrane of such an endovesicle can be characterized by its negative spontaneous curvature, which may lead to a torocyte endovesicle shape with a small relative volume. Effects of a possible strong interaction between the C12E8-induced membrane inclusions and the membrane on the stability of the torocyte endovesicles are also indicated

The antiparasitic compound Licochalcon A is a potent echinocytogenic agent that modifies erythrocyte membrane in the concentration region where the antiplasmodial activity is observed

The well-known antiparasitic compound licochalcone A is a potent membrane-active agent that transforms normal erythrocytes into echinocytes in parallel with the inhibition of growth of Plasmodium falciparum cultures, the in vitro antiplasmodial effect apparently being an indirect effect on the host cell. In vitro experiments with synchronous cultures demonstrate that inhibition of invasion is the principal mechanism of growth inhibition. The erythrocyte membrane-modifying effect was also transiently observed in vivo in mice after intravenous administration

Spontaneous curvature of ganglioside GM1 -Effect of cross-linking

a b s t r a c t The membrane-curvature dependent lateral distribution of outer leaflet ganglioside GM1 (GM1) and the influence of GM1 cross-linking induced by fluorophore-tagged cholera toxin subunit B (CTB) plus anti-CTB was analysed in cell membranes by fluorescence microscopy. Data are presented indicating that cross-linked GM1-ligand patches accumulated at the tips of human erythrocyte echinocytic spiculae induced by Ca 2+ /ionophore A23187. However, when lipid fixative osmium tetroxide was added prior to the ligand no accumulation in spiculae occurred. GM1-staining remained here distributed over the spheroid cell body and in spiculae. Similarly, osmium tetroxide completely prohibited CTB plus anti-CTBinduced GM1 patching in representatives for flat membrane, i.e. discoid erythrocytes and K562 cells. Our results demonstrate that GM1 per se shows low membrane curvature dependent distribution and therefore holds flexible spontaneous curvature. In contrast, the cross-linked GM1-ligand complex has a strong preference for highly outward curved membrane and possesses overall positive spontaneous curvature. Osmium tetroxide efficiently immobilises GM1

Minimizing isotropic and deviatoric membrane energy – An unifying formation mechanism of different cellular membrane nanovesicle types

Tiny membrane-enclosed cellular fragments that can mediate interactions between cells and organisms have recently become a subject of increasing attention. In this work the mechanism of formation of cell membrane nanovesicles (CNVs) was studied experimentally and theoretically. CNVs were isolated by centrifugation and washing of blood cells and observed by optical microscopy and scanning electron microscopy. The shape of the biological membrane in the budding process, as observed in phospholipid vesicles, in erythrocytes and in CNVs, was described by an unifying model. Taking the mean curvature h and the curvature deviator d of the membrane surface as the relevant parameters, the shape and the distribution of membrane constituents were determined theoretically by minimization of membrane free energy. Considering these results and previous results on vesiculation of red blood cells it was interpreted that the budding processes may lead to formation of different types of CNVs as regards the compartment (exo/endovesicles), shape (spherical/tubular/torocytic) and composition (enriched/depleted in particular kinds of molecules). It was concluded that the specificity of pinched off nanovesicles derives from the shape of the membrane constituents and not primarily from their chemical identity, which explains evidences on great heterogeneity of isolated extracellular vesicles with respect to composition.</p

Prevalence of stocked whitefish in River Kemijoki, Finland, inferred by micro X-ray fluorescence analysis of otoliths

Micro X-ray fluorescence (mu-XRF) analysis of otoliths was evaluated as a method to estimate the proportion of stocked one-summer-old whitefish Coregonus lavaretus L. in catches of adult fish (n = 20) ascending the River Kemijoki to spawn. Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) analysis was applied as control. Polished otoliths were scanned with mu-XRF to obtain strontium maps that were used to infer visually the provenance of the whitefish. Thirteen of the fish showed signs of being stocked as one-summer-old fingerlings. LA-ICP-MS was applied to determine the elemental composition in a spot outside the core of the otolith. The results were largely consistent with the visual inspection of the mu-XRF mapped otoliths. In conclusion, mu-XRF mapping successfully identified whitefish stocked as one-summer-old fingerlings. The vast majority of whitefish returning to the River Kemijoki to spawn were stocked fish

Membrane skeleton and red blood cell vesiculation at low pH

AbstractShapes of red blood cells at low pH were studied theoretically. It is assumed that the equilibrium shape of the red blood cell corresponds to the minimum of its membrane elastic energy which consists of the bending energy and relative stretching energy of the bilayer, the stretching energy of the skeleton and the interaction energy between the skeleton and the bilayer. It is shown that the aggregation of the skeleton at low pH can cause the red blood cell shape transformation from the stomatocytic shape to the cell shape composed of a spherical parent cell having the bilayer completely underlaid with the skeleton and spherical daughter vesicles without the skeleton

Proportion of river- and sea-spawning whitefish in catches at the Åland Islands (Baltic Sea), estimated from gill raker counts

The Åland Islands are feeding grounds for river- and sea-spawning whitefish (Coregonus lavaretus), including also hatchery-reared released whitefish. Management of fisheries requires knowledge of whitefish origin from their catches, but such information is deficient at the Åland Islands. The numbers of gill rakers had unique (discretized) Gaussian frequency distributions in all three forms studied. This justified the usage of Gaussian finite mixture models on gill raker data for estimating the proportions of the forms in mixed catches during the non-spawning season. From 14 catches at the Åland Islands from 2012–2013, on average 54.8% were river-spawning whitefish, while the presence of hatchery-reared, sea-spawning whitefish was low but uncertain. Using a hierarchical variant of the model, we showed that the proportion of river-spawning whitefish decreased from 69.6% in June to 38.3% in August. Our study provides feasible methods for monitoring spatio-temporal variation in the proportions of river- and sea-spawning whitefish