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Y Marikovsky

Publications and source records attributed to Y Marikovsky.

At least 19 recordsLinked to original sources
PubMed1990

Phospholipid lamellae are cholesterol carriers in human bile.

Cholesterol solubility and precipitation in bile are major factors in the pathogenesis of cholesterol gallstones. At present, mixed micelles and phospholipid vesicles are considered to be the only cholesterol carriers in bile. In this study we present evidence showing that phospholipid lamellae are major cholesterol carriers in human bile. Lamellae are a known aggregational form in pure phospholipid model systems. In the present study, lamellae were demonstrated by electron microscopy after negative staining and by small-angle X-ray diffraction in all human gallbladder bile samples examined. During diffraction experiments, cholesterol was found to crystallize from these lamellae. Cholesterol carriers in bile were separated by high-resolution chromatography and by prolonged ultracentrifugation. Lamellae were shown to solubilize most of the biliary cholesterol; vesicles solubilized a lesser amount; while micelles solubilized only a minor portion. Our data suggest that phospholipid aggregates are the main cholesterol carriers in bile. Bile salts may control the equilibrium between the various aggregational forms of cholesterol-carrying phospholipids.

Bile↗
PubMed1989

Sequestrocytes: a manifestation of transcellular cross-bonding of the red cell membrane in sickle cell anemia.

An unusual form of red cells (called sequestrocytes) that circulate in the peripheral blood of patients with sickle cell anemia has been identified. In wetmount light microscopy preparations, sequestrocytes appear as massively vacuolated erythrocytes. We have shown by transmission electron microscopy and scanning electron microscopy that the morphology of sequestrocytes can be accounted for on the basis of transcellular cross-bonding of the cell membrane. Initially, endocytotic vesicles span the cytoplasmic space, forming putative cytoskeletal fusion zones. Points of fusion of the membrane skeleton bridging the cytoplasmic compartment expand laterally to form linear fusion zones that entrap lakes of hemoglobin within pseudovacuoles. Sequestrocytes can be isolated in the densest layer (1.159 g ml-1 or greater) of an arabinogalactan density gradient. These cells can be generated in increased numbers in sickle cell blood by incubating samples in 1.5 mM-acetylphenylhydrazine (APH) solution for two hours at 37 degrees C. Their formation is partially blocked by incubation with the reducing agent, dithiothreitol (DTT). Our results suggest that these cells represent an expression of oxidative membrane injury in sickle cell anemia.

Adult↗
PubMed1988

Reconstitution of fusogenic Sendai virus envelopes by the use of the detergent chaps.

Sendai virus envelopes have been a useful tool in studying the mechanism of membrane-membrane fusion and have served as a vehicle for introducing foreign molecules (e.g., membrane proteins) into recipient cells. Reconstituted Sendai virus envelopes are routinely obtained following solubilization of virus particles with Triton X-100. This detergent has a low critical micellar concentration which precludes it from being the best detergent of choice in reconstitution studies. Nevertheless, it has remained in use since other detergents such as sodium deoxycholate and sodium cholate rendered the resultant vesicles inactive. Triton X-100 may be suboptimal for studies of some proteins that need be coreconstituted with the viral envelopes. Thus, alternative advantageous detergents, which retain the envelope fusogenic activity, have been sought. In this study we show that the synthetic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (Chaps) effectively solubilizes the Sendai virions, and that the vesicles formed by simple reconstitution protocols appear structurally and biochemically similar to those obtained with Triton X-100. The resultant vesicles retain functional integrity as assessed in both fusion and hemolysis assays. This protocol seems to be useful in sendai envelope-mediated reimplantation of Fc epsilon receptors into the plasma membranes of rat basophilic leukemia cells.

Cell Membrane↗
PubMed1988

Variations in surface charge distribution of leukemic and non-leukemic transformed cells.

The surface negative charge distribution of fresh leukemic cells from patients with acute leukemia was examined with cationized ferritin (CF), an electron dense label of anionic sites and compared with leukemic cells from phase-cycle fractionated populations of a human leukemic cell line K-562. Normal-untreated and phytohemagglutinin (PHA)-transformed lymphocytes were also tested. A CF-induced redistribution of anionic sites into clusters and patches within a wide range of variations in density was observed electron microscopically both in fresh acute leukemic cells and in unseparated cells of the K-562 line. Cells of the G0/G1 fraction from a phase-cycle separated K-562 population exhibit a higher density of CF label per unit length of membrane as compared with the G2 + M cell fraction. PHA-benign transformed normal lymphocytes exhibit an even and continuous CF labeling, similar to that of normal-untreated lymphocytes. An attempt was made to correlate the CF-induced charge redistribution with the rate of agglutination with the cationic Poly-L-lysine. Our observations indicate that the CF ligand does induce an anionic site redistribution, varying in density, both in fresh leukemic cells and in cells from a K-562 line, and does not in normal untreated and PHA-benign transformed lymphocytes. The particular pattern of anionic site redistribution observed in leukemic cells of this study is apparently phase-cycle dependent.

Acute Disease↗
PubMed1988

The aging of the red blood cell. A multifactor process.

Red blood cell (rbc) senescence is associated with loss of surface sialic acid, which is the principal carrier of surface negative charge and determines the electrokinetic behavior of old rbcs. Loss of sialic acid in an old rbc is demonstrated in its decreased electric mobility and lower negative charge density, determined topographically with cationic particle labeling. Surface sialic acid determines also the mutual attraction--repulsion forces, as demonstrated in enhanced aggluinability with cationic molecules, lectins, and blood group antibodies. Loss of sialic acid accompanies ATP-depletion in vitro; thus, a T-antigen site is unmasked. Macrophages have specific receptors to the site as to newly exposed galactose and N-acetyl galactosamine sugars. Furthermore, the involvement of complement molecules in the recognition of old RBCs by macrophages has been shown. This is possibly due to loss of sialic acid or at least a regrouping--relocation of surface anionic sites due to cell shape changes from discocytes to crenated forms, which accompany both in vivo and in vitro rbc aging. In turn, shape changes are apparently controlled by the cytoskeletal network underlying the rbc membrane, which undergoes structural alteration with physiologic aging in changing the dimensions of oligomeric spectrin and the thickness of the spectrin-actin cytoskeletal assembly.

Animals↗
PubMed1987

Ethanol-induced alterations in human erythrocyte shape and surface properties: modulatory role of prostaglandin E1.

Exposure of human erythrocytes to ethanol (1 to 20% by vol) in Ca2+ and Mg2+-free phosphate-buffered saline, pH 7.4, transformed biconcave discs into spiculated echinocytes within 3 min at 25 degrees C. The effects of ethanol were concentration-and time-dependent, but reversible by washing in the incubation buffer system within 60 min of initial exposure to ethanol. After prolonged ethanol exposure (180 min), washing of cells resulted in the formation of stomatocytes (cup-forms). Ethanol-induced echinocytosis was also accompanied by a 30% enhancement in the agglutinability of erythrocytes by ligands with high affinity for negative surface charge (poly-L-lysine and wheat germ agglutinin, 20 microliters/ml) without any alterations in surface charge topography. Concomitant exposure of erythrocytes to prostaglandin E1 (100 nM) selectively prevented the enhancement of ligand-mediated agglutinability, but did not modify cell shape. These data indicate that certain erythrocyte surface properties may not be directly influenced by cell shape and suggest a unique modulatory action of prostaglandin E1 on shape-transformed cells.

Alprostadil↗
PubMed1987

Modulation of polycation-induced redistribution of melanoma cell surface anionic macromolecules by retinoic acid.

The ability of cationized ferritin (CF) to redistribute negatively charged cell surface molecules has been shown to increase after malignant transformation. Pretreatment of murine melanoma S91-C2 and B16-F1 cells with retinoic acid (RA), which suppresses their transformed phenotype, decreased the ability of CF to cluster surface anionic sites. In contrast, a similar pretreatment of RA-resistant mutant clone S91-C154 and subline B16-F10 caused only a minor reduction in CF-induced patching of anionic sites. These results indicate that the effect of RA on the redistribution of negatively charged cell surface molecules is related to the growth-inhibitory action of this vitamin A metabolite.

Animals↗
PubMed1986

Distribution and modulation of surface charges of cells from human leukemia-lymphoma lines at various stages of differentiation.

Untreated and retinoic acid (RA) treated human leukemia-lymphoma cell lines reflecting hematopoietic cells at various stages of differentiation, were examined electron microscopically for their surface negative charge distribution using cationized ferritin (CF), an electron dense label of anionic sites. The results indicate that there is a correlation between the CF labeling density/distribution and the stage of lymphoid cell differentiation. Viable unfixed null cell lines show a low CF labeling density with few and small CF patches. A gradual increase in CF labeling density and increase in size and number of CF patches correlates with the stage of differentiation on cell lines of both T or B origin. Treatment of viable unfixed cells with 10(-5) MRA for 10 days seems to prevent the CF-induced formation of CF patches, resulting in a continuous and even distribution of the CF label, similar to that observed on the surface of cells fixed before CF labeling. Some correlation between the distribution of surface anionic sites and the malignant potential of the human leukemic lines could be detected.

Anions↗
PubMed1986

Cholesterol-phospholipid vesicles in human bile: an ultrastructural study.

Phospholipid vesicles, a newly described (bile salt independent) mode of cholesterol transport in human bile, were previously characterized by quasi-elastic light scattering and gel filtration. In the present study the ultrastructure of these vesicles was investigated by electron microscopy using freeze-fracture and negative-staining techniques. Vesicles of varying size were found in all 14 hepatic and 3 gallbladder biles examined. The diameter of the vesicles ranged from 25 to 75 nm by electron microscopy after freeze fracture and from 54 to 94 nm by quasi-elastic light scattering. They had a spherical shape and appeared to be unilamellar. The appearance of the vesicles in fresh hepatic and gallbladder biles as well as in chromatographic fractions was similar. Vesicles were dissolved by the addition of exogenous bile salts. Cholesterol is transported in human bile by both vesicles and micelles. The role of the vesicles may be particularly important in preventing cholesterol precipitation in dilute and supersaturated biles.

Bile↗
PubMed1985

The relationship between sialic acid content and peanut agglutinin binding on senescent and enzyme treated human erythrocytes.

Young, old and neuraminidase treated human red blood cells (RBC) were investigated with peanut agglutinin (PNA), a lectin with a specificity similar to that of serum T-agglutinin. The effect of serum agglutinins on this interaction was also investigated. The density and distribution of PNA receptors were evaluated by agglutination with PNA and binding of ferritin-conjugated PNA (PNA-F), or PNA labeled with radioactive iodine [( 131I] PNA). The results were correlated with the distribution of membrane bound sialic acids, as evaluated by chemical analysis and rate of agglutination with poly-L-lysine (PLL). Untreated RBC of all ages did not agglutinate with PNA and failed to bind PNA-F and [131I] PNA. Treatment of young RBC with neuraminidase, which resulted in reduction of membrane-bound sialic acids to an extent similar to that of physiologically aged RBC, resulted in the concomitant exposure of PNA binding sites and in the agglutination of these cells by autologous serum. Pretreatment of the neuraminidase treated RBC with autologous serum resulted in partial inhibition of the binding capacity of PNA on the RBC. The results indicate that the normal age-related loss of sialic acids in circulating RBC is not identical with enzymatic removal of sialic acids by neuraminidase. The observations suggest that different mechanisms are functional in the recognition and sequestration of old RBC and of RBC treated with neuraminidase.

Erythrocyte Aging↗
PubMed1985

Changes of cell shape and surface charge topography in ATP-depleted human red blood cells.

ATP depletion crenates human red blood cells. With ferritin-avidin (FA) and cationized ferritin (CF) cell surface labeling, it is demonstrated that the discocyte----crenated shape transformation alters the two-dimensional topography of negative charge sites. With restoration of ATP levels, cell shape and charge topography return to normal. Concurrent changes in red cell shape and surface charge topography can be explained by associations between membrane integral proteins and the red cell cytoskeleton.

Adenosine Triphosphate↗
PubMed1985

Surface charge distribution in normal and transformed rat bladder epithelial cells in vitro.

Cationized ferritin (CF) was used as an ultrastructural marker to study differences in the distribution and density of surface anionic charges between normal and neoplastic cells. In anchorage-independent cell systems, CF induces a redistribution of cell surface receptor-ligand complexes into clusters, patches, and caps on the surfaces of transformed cells, but not on the surfaces of normal cells. In the present report, the authors have, for the first time, extended the CF labeling studies to a system of anchorage-dependent, rat bladder epithelial cell lines: normal RBTC cells and RBTCC-8 carcinoma cells. Cells were grown to confluency and labeled with 500 micrograms/ml CF for 3 minutes. After completion of CF labeling, cells were incubated for 0, 15, 60, 120 minutes, or overnight and were fixed then with buffered glutaraldehyde for electron microscopy. The results show that (a) CF induces grouping of surface anionic charge sites into patches and clusters on RBTCC-8 carcinoma cells (CF covers 53.86 +/- 2.15% of the plasma membrane), but not on normal RBTC cells (CF covers the entire plasma membrane); (b) CF densities are well correlated to the cell surface sialic acid contents of RBTC (2.42 +/- 0.30 microM/10(9) cells) and RBTCC-8 cells (1.26 +/- 0.25 microM/10(9) cells); and (c) CF-labeled membrane is internalized at equal rates by RBTC and RBTCC-8 cells (30% in 15 minutes; 60% in 60 minutes; 70% in 120 minutes; 100% overnight). These data indicate that CF-induced patching of anionic sites is a surface characteristic that is common to anchorage-dependent and independent tumor cells. Mechanisms of CF patching in neoplastic cells are discussed.

Animals↗
PubMed1984

Liposome-mediated transfer of macromolecules into flagellated cell envelopes from bacteria.

We have studied the interaction between flagellated cell envelopes from Escherichia coli and liposomes. Oligolamellar liposomes of ca. 0.45-micron diameter, composed of azolectin, phosphatidylserine, and cholesterol at a molar ratio of 7:1:2, were prepared by freezing and thawing and subsequent extrusion through polycarbonate filters. These liposomes exhibited high entrapment capacity and low leakiness. Liposome-cell envelope interaction was monitored flow cytometrically in a fluorescence-activated cell sorter with a fluorescent aqueous space marker and by a filtration assay with radiolabels for the lipid phase and the liposomal aqueous space. Maximal association of liposomes with the envelopes was observed in both assays after ca. 25 min at 30 degrees C. After such period of time, it seems that up to 200 liposomes (depending on the liposome to envelope ratio) were associated with a single cell envelope, as calculated from the radiotracer studies. Fluorometric measurements of the transfer of liposomal contents and the intermixing of membrane lipids indicated that at least 20% of the envelope-associated liposomes had delivered their content into the envelopes, possibly by fusion. Electron microscopic observations confirmed the transfer of liposome-encapsulated ferritin molecules into the cell envelopes. Our data suggest that liposomal carriers might be employed to deliver cytoplasmic, chemotaxis-related macromolecules into bacterial cell envelopes.

Animals↗
PubMed1984

Prevention by retinoic acid of anionic site redistribution on the surface of cultured human sarcoma cells.

The distribution of cell surface negatively-charged macromolecules was determined electron microscopically on untreated and on retinoic acid (RA)-treated cultured human osteosarcoma Hs791 and chondrosarcoma Hs705 cells using cationized ferritin (CF), an electron-dense marker of anionic sites. Labeling on the surface of prefixed cells was continuous and uniform whether they were grown in the absence or presence of RA. In contrast, CF distribution on unfixed cells was markedly affected by RA; CF labeling of untreated cells occurred in patches and clusters whereas the label on RA-treated cells was continuous, as on prefixed cells. CF labeling of unfixed cells decreased considerably after incubation of the cells either with hyaluronidase or neuraminidase. There was also a reduction in patching and clustering. Changes induced by RA in the apparent membrane microviscosity, in neuraminidase-releasable sialic acid, or in transglutaminase activity could not be related to the effect of RA on CF-induced anionic site redistribution since these characteristics were modulated differently in the two cell lines. In contrast, RA increased the sialylation of specific cell surface membrane glycoproteins on both cell types. These results suggest that RA prevents redistribution of cell surface sialoglycoconjugates and glycosaminoglycans by CF. This effect may be the result of increased sialylation of specific surface components and may be related causally to the suppression of the transformed phenotype in the sarcoma cells.

Acyltransferases↗
PubMed1983

Distribution of surface anionic sites on mouse hybrid myelomas.

The surface anionic site distribution on membranes of a monoclonal antibody-producing hybridoma cell line, and its two parental cells: normal spleen cells of immunized BALB/c mice and cells from a mouse myeloma line (NS-1), were investigated with the aid of the cationized ferritin (CF) labelling method, following glutaraldehyde/formaldehyde fixation of cells. The patch-like CF distribution on the hybridoma cells is similar to that of the NS-1 myeloma cells, but distinct from the even and continuous CF distribution of the immunized and nonimmunized normal spleen lymphocytes. The similarity in the formation of patch-like CF heaps, both on myeloma and hybrid cells is discussed in respect to the surface charge characteristic determined by cell fusion.

Animals↗