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PubMed1992

Sterically stabilized liposomes.

Many recent reports have demonstrated that rapid uptake of liposomes in vivo by cells of the mononuclear phagocytic system (MPS), which has restricted their therapeutic utility, can be overcome by incorporation of lipids derivatized with the hydrophilic polymer polyethylene glycol (PEG). The structure-function relationship of PEG-derivatized phosphatidylethanolamine (PEG-PE) has been examined by measurement of blood lifetime and tissue distribution in both mice and rats. The results are reviewed and contrasted with those from liposomes without PEG-PE or other surface modifications. With a PEG molecular weight in the range of 1000 to 5000, prolonged circulation and reduced MPS uptake is achieved. After 24 h, up to 35% of the injected dose remains in the blood and less than 10% is taken up by the two major organs of the MPS, liver and spleen, compared with 1% and up to 50%, respectively, for liposomes without PEG-PE. Other important advantages of PEG-PE have been identified: prolonged circulation is independent of liposome cholesterol content, degree of hydrocarbon chain saturation in either the PC or the PE lipid anchor, lipid dose, or addition of most other negatively charged lipids. This versatility in lipid composition and dose is important for controlling drug release in a liposome-based therapeutic agent. Steric stabilization has been proposed as a theoretical basis for the results and some initial results testing this hypothesis have been reported. A description of a theoretical model is presented here and evaluated with the data available. The results are compared with other particulate drug carriers and the range of potential applications are considered.

Animals↗
PubMed1977

Distribution of anionic sites on the oviduct ciliary membrane.

Polycationic ferritin (PCF) was used as a visual probe for anionic sites on the oviduct ciliary membrane. The binding of PCF to ciliary membranes was dependent on the concentration of the probe in the incubation media. At low concentrations (0.08-0.16 mg/ml), PCF was bound exclusively to the tip of the cilium whereas at higher concentrations (0.32-0.64 mg/ml), ferritin was located at the tip and at the base around the transition region, with occasional scattered clumps on the remainder of the membrane. The base and tip binding was fount to be associated with special surface modifications of the membrane in these regions. At the tip, PCF was bound to a filamentous glycocalyx termed the ciliary crown. Base binding was associated with a system of five to six 140-A high ridges, each of which encircled the membrane of the transition region. The ridges were equally spaced (approxamately 245 A spacing) along the length of the transition region. Since pretreatment of oviduct with either neuraminidase or protease blocked the binding of the probe, the PCF-binding sites appear to be negatively charged glycoproteins or mucopolysaccharides.

Animals↗
PubMed1976

Ultrastructural studies of surface features of human normal and tumor cells in tissue culture by scanning and transmission electron microscopy.

Human tumors of a variety of histopathologic types have been established in tissue culture. The surface features of these cell lines were investigated by scanning electron microscopy (SEM) with the use of new techniques for specimen preparation. Tumor cells demonstrated striking degrees of surface activity with numerous microvilli, filopodia, blebs, and ruffles. Intercellular contacts were also prominent in cultures of most solid tumors observed by SEM. At low cell density, normal human fibroblasts exhibited some surface features such as microvilli and blebs, but at higher cell density they lacked extensive surface modifications. By transmission electron microscopy (TEM), the cytoskeleton of normal fibroblasts was shown to be well organized, with parallel orientation of microfilaments, filaments, and microtubules. These structures were also in tumor cells, but they lacked the degree of organization of fibroblasts. Desmosomes were readily demonstrated in normal fibroblasts and carcinoma cells in culture but not in sarcomas, melanomas, or tumors of neural origin. These studies have provided the first correlative SEM and TEM analyses of solid human tumor cells of diverse pathologic types in vitro.

Carcinoma↗
PubMed1979

Adaptive differences of the surface organization of horny cells.

The existence of a peculiar structural pattern of complementary villi and pores was observed on the squamous surfaces of palms and soles. These features, which might be considered as acquired and adaptive cell surface modifications, proved to be more conspicuous during a period of rat and human postnatal development.

Animals↗
PubMed1979

The role of sialated glycoproteins in endocytosis, permeability and transmural passage in the myeloid endothelium.

Changes in the anionic charge distribution at the luminal face of the endothelium of the sinusoids of the bone marrow have been studied at sites of endocytosis by large bristle coated vesicles and at the sites of molecular permeability through diaphragmed fenestrae. The anionic charge distribution has also been studied at the abluminal aspect of these vessels at sites of transmural blood cell passage. Cationic surface markers such as colloidal iron, native ferritin and polycationic ferritin used at low pH, 1.8, and the use of neuraminidase show that the nonmodified endothelial cell surface has exposed sialic acid groups, which are absent at the sites of these functional specializations. Polycationic ferritin binding over a range of pH levels indicates the prsence of another species of anionic materials present at both the nonmodified cell surface and at the sites of the cell surface modifications. This second group of anionic compounds is neuraminidase resistant and has a pKa higher than that of sialic acid (pKa:2.6).

Binding Sites↗
PubMed1992

Block copolymers as drug carriers.

The main subject of this review is to describe the concept and strategy of utilizing block copolymers as drug carriers and to present a perspective for this utilization. The first section points out preferable properties of block copolymers as drug carriers with a brief survey on classifications and synthesis of block copolymers and specifies the merits of block copolymers for this purpose in the historical background of polymeric drug carriers. The second section introduces several studies using block copolymers as drug carriers. These studies contain surface modification of microspheres with block copolymers, polymeric micelles, and a conjugate of an antibody and block copolymer. In conclusion, it is determined that block copolymers possess a high potential for use as drug carriers irrespective of some difficulties in their synthesis.

Animals↗
PubMed1979

An ultrastructural comparison of human endometrial adenocarcinoma with normal postmenopausal endometrium.

Although electron microscopy has been of limited value in detecting malignant neoplasms, some neoplasms do exhibit characteristic ultrastructural features. Intensive study of these features may offer insight into the etiology and activity of such tumors. In an attempt to characterize the ultrastructure of endometrial adenocarcinoma, tissue was examined with the electron microscope and compared with normal postmenopausal endometrium. Endometria from biopsy, hysterectomy, or curettage were processed routinely for light and electron microscopy. Several ultrastructural features of the adenocarcinoma were common both to previous descriptions of endometrium of the postovulatory phase and to the normal postmenopausal endometrium, described here, viz, atypical mitochondrial forms and cell surface modifications. Ribosomes were abundant in adenocarcinoma of the endometrium, normal postmenopausal endometrium, and normal cyclic endometrium in the preovulatory phase.

Adenocarcinoma↗
PubMed1979

Extraskeletal Ewing's sarcoma. Histologic and ultrastructural observations in three cases.

The histologic and ultrastructural morphology of three cases of Ewing's sarcoma of soft tissue are described and the fine structural features of extraskeletal Ewing's sarcoma are compared to those of similar round-cell tumors that are considered in the differential diagnosis. By light microscopy, these tumors are indistinguishable from Ewing's sarcoma of bone. Ultrastructurally, the salient features are also comparable to Ewing's sarcoma of bone and include: 1) absence of surface modifications; 2) cell-contact sites in the form of small thickenings of apposed membranes and large desmosome-like specializations; 3) undifferentiated cytoplasm usually containing abundant glycogen and occasionally nonspecific microfilaments; 4) significant variation in shape and irregularity of nuclear profiles. The ultrastructural features of extraskeletal Ewing's sarcoma are sufficiently distinctive to allow separation from from other small-cell malignant neoplasms in the majority of cases.

Adult↗
PubMed1978

The structure of the germinal disc region of the hen's ovarian follicle during the rapid growth phase.

The structure of the ovarian follicle in the region of the germinal disc, which appears as a white plaque at the surface of the oocyte, was examined by electron microscopy and compared with the non-disc region which overlies the yellow yolk mass of the oocyte in the final growth phase. The main differences concerned the granulosa cell layer and the surface layer of the oocyte. In the disc the granulosa cells were less regularly arranged and the spaces between them varied in width. Their mitotic rate was higher than that in the non-disc region, where cell division was seldom observed at maturity. The perivitelline layer was comparatively poorly developed at the periphery of the germinal vesicle in 15 mm follicles, but eventually attained a uniform thickness throughout the follicle. In the intercellular and perivitelline spaces there were smaller amounts of granular material. Marked differences were observed in the ooxyte surface layer. In 15 mm follicles the surface of the germinal disc was thrown into numerous microvilli and some narrow indentations containing macrovilli from the granulosa cells. Coated vesicles, 120 nm diameter, appeared to be invaginating from the oolemma, whereas 70 nm coated vesicles were present in the deeper cytoplasm. In follicles of more than 25 nm diameter these structural conformations were evident only at the periphery of the disc; for the most part the 120 nm coated vesicles were absent, and over the germinal vesicle microvilli were of rare occurrence. On the other hand, the bulk of the oocyte surface was highly convoluted throughout this period of growth, numerous granulosa cell macrovilli extended into deep pouches associated with 300 nm coated vesicles, and the oolemma possessed a coating of fuzzy material. These observations suggest that there is a restricted passage of yolk precursors to the surface of the germinal disc, and that the inability to transport yellow yolk into the disc is related to differences in the oolemmal surface coat and the population of coated vesicles. The surface modifications, as well as the proliferation of the granulosa cells, are likely to be influenced by the presence of the germinal vesicle.

Animals↗
PubMed1976

Platelets, foreign surfaces, and heparin.

These studies clearly indicate that heparin has 2 antagonistic effects in this platelet-foreign surface interaction; it acts directly on platelets to increase retention, while acting on the foreign surface to reduce platelet retention, perhaps by competing for cationic sites. This study also suggests that it is unlikely that heparin bonding confers reduced surface thrombogenicity by virtue of the ability to leach off that surface and that further studies in heparin bonding or other surface modification should consider the antiplatelet effect which can easily be studied in vitro.

Blood Platelets↗
PubMed1977

Effect of chemical modification of cell surface components of a brewer's yeast on the floc-forming ability.

Effects of treatments with proteolytic enzymes and protein-modifying reagents on flocculation of brewer's yeast IFO 2018 were investigated. The floc-forming ability of the yeast cells was irreversibly eliminated by treatment with papain, trypsin, chymotrypsin or pepsin, indicating that certain proteins on the cell surface participate in the yeast flocculation. Chemical modification with reagents, known to act on disulfide bridges, carboxyl and/or phosphate groups, phenolic groups, amino groups, and imidazole groups, also destroyed the ability to flocculate, although in some cases a high concentration (8 M) of urea was necessary in addition to protein-modifying reagents. Thus, it is suggested strongly that these functional groups of amino acid residues of the proteins are essential for the floc-forming ability of brewer's yeast cells.

Cell Wall↗
PubMed1975

Cholera toxin effects on cell growth accompanied by selective alterations in metabolite uptake and modification of cell surface proteins.

Exposure of Chinese hamster ovary cells to cholera toxin at or below mug levels causes a marked morphological changes and increased adhesion and orientation of the cells. Such changes are paralleled by alterations in surface proteins as indicated by the cholera toxin-mediated modifications detectable by lactoperoxidase-catalyzed radioiodination of outer proteins. Mild tryptic treatment of cells prelabeled with [3H]glucosamine revealed a different kinetics of release of external glycoproteins in cells exposed to the toxin. An alteration in a specific glycoprotein species in cholera toxin-treated cells became evident by polyacrylamide gel electrophoresis followed by fluorography of 3H-labeled cellular glycoproteins. The effects of cholera toxin on surface proteins and growth of the cells occurred in the absence of a modification in amino acid uptake or incorporation of precursors into protein. However, thymidine uptake and glucosamine incorporation were inversely affected to toxin treatment. Some of the effects of the toxin appeared to be antagonized by colchicine.

Amino Acids↗
PubMed1977

A comparative study of the effect of modification of the surface of human platelets on the receptors for aggregated immunoglobulins and for ristocentin-von Willebrand factor.

The receptors for aggregated immunoglobulin G (IgG) (an Fc receptor) and for ristocetin-von Willebrand factor on human platelets were studied by means of various modifications of the platelet surface. The expression of these receptors was measured by the agglutination of platelets to ristocetin in the presence of von Willebrand factor, which is part of the factor VIII complex, and by the binding of aggregated IgG coupled to 3H-labelled diazobenzene. Treatment of platelets with chymotrypsin, trypsin, papain and pronase which removed protein and glycoprotein from the platelet under conditions where the release reaction was inhibited caused loss of the expression of the receptor for ristocetin-von Willebrand factor and an enhancement of that for aggregated IgG. Induction of membrane changes with ADP and of the release reaction with the ionophore A23187 abolished agglutination to ristocentin-von Willebrand factor but did not alter the receptor for aggregated IgC. Possible contributions of unspecific membrane changes, produced by protease treatment of platelets, to the modification of receptor expression were eliminated by the use of formaldehyde-treated platelets. Trypsin, papain and pronase destroyed the ability of these platelets to agglutinate to ristocetin-von Willebrand factor but produced no change in the binding of aggregated IgC. Therefore, the receptor for ristocetin-von Willebrand factor is truly sensitive to proteolysis while the Fc receptor is not, but is partially masked by protease-sensitive material.

Adenosine Diphosphate↗
PubMed1976

Selective modification of cell surface proteins and thymidine transport in hamster cells exposed to cholera toxin.

The increased adherence and morphological response which occurs in Chinese hamster ovary cells as a result of exposure to cholera toxin is paralleled by modification in the relative exposure of outer proteins. Mild proteolysis treatment of the cells prelabeled with [3H] glucosamine reveals a markedly different kinetics of release of external glycopeptides as a result of exposure to cholera toxin. Selective alterations in external tyrosyl-rich proteins can also be detected by lactoperoxidase-catalyzed radioiodination. The above modifications are accompanied by a decrease in the rate of thymidine uptake by toxin-treated cells.

Biological Transport, Active↗
PubMed1978

Chemical modification of the surfaces of bacterial cell walls.

The surfaces of the isolated cell walls of four bacterial species were studied by microelectrophoresis following chemical treatments intended to remove specific charged groups. Acid-base titrations of the walls were used to assess specificity and extent of the modifications. Carboxyl groups were specifically and completely modified by activation with a water-soluble carbodiimide and subsequent reaction with a nucleophile, such as glycinamide, to give an uncharged pH-stable product. Aqueous media and mild reaction conditions make the method suitable for modifying carboxyl groups on cell surfaces too labile to withstand the harsh conditions required for conventional esterification reactions. Use of the carbodiimide-mediated reaction for discharging carboxyl groups, along with fluorodinitrobenzene for discharging amino groups and extraction procedures for removing constituents carrying phosphoester groups (teichoic acids), made it possible to obtain information about the spatial arrangement of charged groups on the wall surfaces. Removal of the exterior negative charge dominating wall surfaces allowed underlying amino groups to become electrokinetically effective and, in the case of E. coli, also revealed a lipophilic region with an affinity for a cationic surfactant.

Bacillus↗
PubMed1992

Nuclear magnetic relaxation dispersion and 31P-NMR studies of the effect of covalent modification of membrane surfaces with poly(ethylene glycol).

Covalent attachment of methoxypoly(ethylene glycol) (MPEG) 5000 to the surface of unilamellar liposomes composed of egg phosphatidylcholine and dioleoylphosphatidylethanolamine (DOPE) (8:2) containing paramagnetic chelates, either entrapped within the interior volume of the liposomes, or associated with the membrane surface, had no effect upon the measured spin-lattice relaxation rates (1/T1) for water in these systems. 31P-NMR studies indicate no destabilization of dioleoylphosphatidylcholine (DOPC)/(DOPE) (1:1) vesicles following attachment of MPEG. However, in DOPC/DOPE (1:3) mixtures, covalent modification with MPEG results in a destabilization of multilamellar vesicles into smaller vesicular structures. These results indicate that covalent attachment of poly(ethylene glycol) to liposomal magnetic resonance agents may prove a useful method for increasing their utility as vascular MR agents by extending their lifetime in the circulation, without decreasing the relaxivity of paramagnetic species associated with the liposome, but that the presence of PEG covalently attached to the membrane surface may modify the polymorphic phase behavior of the lipid system to which it is covalently linked.

Lipid Bilayers↗