Isolation and chemical characterization of Alzheimer's disease paired helical filament cytoskeletons: differentiation from amyloid plaque core protein.
AuthorsRoher AE, Palmer KC, Chau V, Ball MJ
JournalJ Cell Biol
PubMed ID3060472
'The paired helical filaments (PHFs) of Alzheimer''s disease were purified by a strategy in which the neurons and amyloid plaque cores of protein (APCP) were initially isolated. This was achieved by several steps of isocratic sucrose centrifugations of increasing molarity and a discontinuous isotonic Percoll density gradient. After collagenase elimination ... More
Electrophysiology of plasma membrane vesicles.
AuthorsWright EM
JournalAm J Physiol
PubMed ID6372509
'In both renal and gastrointestinal physiology, it has become popular to study epithelial transport phenomena using vesicles isolated from the apical and basolateral cell membranes. Transport in vesicle preparations is usually monitored with radioactive tracers, but more recently attention has been directed to electrophysiological methods. As it is impossible to ... More
Na+, K+, H+ and Cl- permeability properties of rabbit skeletal muscle sarcolemmal vesicles.
AuthorsGilbert JR, Meissner G
JournalArch Biochem Biophys
PubMed ID6859866
'The ion permeability properties of rabbit skeletal muscle sarcolemmal vesicles were investigated by means of radioisotope flux, membrane potential, and light-scattering measurements. An enriched sarcolemmal fraction was obtained from the 22-27% region of sucrose gradients after isopycnic centrifugation. The presence of contaminating sarcoplasmic reticulum was assessed with the use of ... More
Ca2+ influx mediates apoptosis induced by 4-aminopyridine, a K+ channel blocker, in HepG2 human hepatoblastoma cells.
AuthorsKim JA, Kang YS, Jung MW, Kang GH, Lee SH, Lee YS
JournalPharmacology
PubMed ID10657756
'Apoptosis appears to be implicated in the pathogenesis and therapeutic applications of cancer. In this study we investigated the induction of apoptosis by 4-aminopyridine (4-AP), a K(+) channel blocker, and its mechanism in HepG2 human hepatoblastoma cells. 4-AP reduced cell viability and induced DNA fragmentation, a hallmark of apoptosis, in ... More
Lateral mobility in membranes as detected by fluorescence recovery after photobleaching.
AuthorsYguerabide J, Schmidt JA, Yguerabide EE
JournalBiophys J
PubMed ID7139035
'The evaluation of lateral diffusion coefficients of membrane components by the technique of fluorescence recovery after photobleaching (FRAP) is often complicated by uncertainties in the values of the intensities F(O), immediately after bleaching, and F(infinity), after full recovery. These uncertainties arise from instrumental settling time immediately after bleaching and from ... More
Relation of transmembrane potential to cell survival following hyperthermia in HeLa cells.
'Hyperthermia induces cell death and the usual endpoint to study this is the ability of the cells to form colonies. Hyperthermia is also known to alter membrane characteristics, especially transmembrane potential and this has been correlated with duration and degree of heating. The aim of the present study was to ... More
Verotoxin 1 binding to intestinal crypt epithelial cells results in localization to lysosomes and abrogation of toxicity.
AuthorsHoey DE, Sharp L, Currie C, Lingwood CA, Gally DL, Smith DG,
JournalCell Microbiol
PubMed ID12580945
'Verotoxins (VTs) are important virulence factors of enterohaemorrhagic Escherichia coli (EHEC), a group of bacteria associated with severe disease sequelae in humans. The potent cytotoxic activity of VTs is important in pathogenicity, resulting in the death of cells expressing receptor Gb3 (globotriaosylceramide). EHEC, particularly serotype O157:H7, frequently colonize reservoir hosts ... More
Effects of deoxycorticosterone/salt hypertension on cell ploidy in rat aortic smooth muscle cells.
AuthorsChobanian AV, Lichtenstein AH, Schwartz JH, Hanspal J, Brecher P
JournalCirculation
PubMed ID3791614
'The effect of hypertension on DNA content of isolated aortic smooth muscle cells and their nuclei has been studied in Wistar rats treated with deoxycorticosterone and salt (DOC/salt). Eight weeks of DOC/salt administration induced a significant increase in aortic smooth muscle cells with tetraploid nuclei from 9.7 +/- 0.8% (mean ... More
Versatile regulation of cytosolic Ca2+ by vanilloid receptor I in rat dorsal root ganglion neurons.
AuthorsLiu M, Liu MC, Magoulas C, Priestley JV, Willmott NJ
JournalJ Biol Chem
PubMed ID12454015
'Analysis of small dorsal root ganglion (DRG) neurons revealed novel functions for vanilloid receptor 1 (VR1) in the regulation of cytosolic Ca(2+). The VR1 agonist capsaicin induced Ca(2+) mobilization from intracellular stores in the absence of extracellular Ca(2+), and this release was inhibited by the VR1 antagonist capsazepine but was ... More
Regulation of capacitative Ca2+ influx in human neutrophil granulocytes. Alterations in chronic granulomatous disease.
AuthorsGeiszt M, Kapus A, Német K, Farkas L, Ligeti E
JournalJ Biol Chem
PubMed ID9334224
'Ca2+ entry through the capacitative (store-regulated) pathway was shown to be inhibited in neutrophil granulocytes by the protein kinase C activator phorbol 12-myristate 13-acetate and the chemoattractant N-formyl-methionyl-leucyl-phenylalanine (fMLP) by a hitherto unknown mechanism. Measuring both Ca2+ and Mn2+ entry into store-depleted cells we show in the present study that ... More
Membrane potential modulates photocycling rates of bacterial rhodopsins.
AuthorsManor D, Hasselbacher CA, Spudich JL
JournalBiochemistry
PubMed ID3191096
'Effects of membrane potential on photochemical reactions of three retinal-containing chromoproteins in Halobacterium halobium, sensory rhodopsin I (sR-I), bacteriorhodopsin, and halorhodopsin, are described. Each of the three exhibits a decreased rate of thermal decay of its principal intermediate when photoactivated in an artificially energized compared to a deenergized membrane. The ... More
Salivary histatin 5 induces non-lytic release of ATP from Candida albicans leading to cell death.
AuthorsKoshlukova SE, Lloyd TL, Araujo MW, Edgerton M
JournalJ Biol Chem
PubMed ID10383383
'Salivary histatins are potent in vitro antifungal proteins and have promise as therapeutic agents against oral candidiasis. We performed pharmacological studies directed at understanding the biochemical basis of Hst 5 candidacidal activity. Three inhibitors of mitochondrial metabolism: carbonyl cyanide p-chlorophenylhydrazone, dinitrophenol, and azide inhibited Hst 5 killing of Candida albicans, ... More
An incremental approach to automated protein localisation.
AuthorsTscherepanow M, Jensen N, Kummert F,
JournalBMC Bioinformatics
PubMed ID18937856
'BACKGROUND: The subcellular localisation of proteins in intact living cells is an important means for gaining information about protein functions. Even dynamic processes can be captured, which can barely be predicted based on amino acid sequences. Besides increasing our knowledge about intracellular processes, this information facilitates the development of innovative ... More
Sensitization to hyperthermia by 3,3'-dipentyloxacarbocyanine iodide: a positive correlation with DNA damage and negative correlations with altered cell morphology, oxygen consumption inhibition, and reduced ATP levels.
AuthorsBorrelli MJ, Rausch CM, Seaner R, Iliakis G
JournalInt J Hyperthermia
PubMed ID1880454
'The cyanine dye 3,3''-dipentyloxacarbocyanine iodide (DiOC5(3)) (concentrations of 0.5 microgram/ml to 5.0 micrograms/ml) was shown to be a potent sensitizer of Chinese hamster ovary (CHO) cells to hyperthermic cell killing at 43.0 degrees C or 45.5 degrees C, while exhibiting no cytotoxicity at 37.0 degrees C. Sensitization to hyperthermic cell ... More
Flow cytometry as an analytic and preparative tool for studies of neuroendocrine function.
AuthorsCambier JC, Monroe JG
JournalMethods Enzymol
PubMed ID6199646
Valinomycin-induced modulation of adriamycin resistance and cationic probe distribution in MCF-7 cell lines.
AuthorsCrifò C, Capuozzo E, Cucco C, Zupi G, Salerno C
JournalBiochem Int
PubMed ID1815496
'In vitro restoration of adriamycin sensitivity in a resistant human breast tumor cell line was obtained by continuous exposure to nanomolar nontoxic valinomycin concentrations. Seven-day treatment with nanomolar valinomycin concentrations caused a slight increase of the signal of the cationic fluorescent cyanine probe DiOC5(3) but did not appreciably affect adriamycin ... More
Flow cytometric probes of early events in cell activation.
AuthorsShapiro HM
JournalCytometry
PubMed ID7023883
Mobile and immobile endoplasmic reticulum in onion bulb epidermis cells: short- and long-term observations with a confocal laser scanning microscope.
AuthorsKnebel W, Quader H, Schnepf E
JournalEur J Cell Biol
PubMed ID2081534
'The endoplasmic reticulum (ER) of onion bulb scale epidermis cells consists of long, tubular strands lying deep in the cytoplasm which move quickly and a less mobile peripheral network of tubules and cisternae that change in position, shape and size but that also have immobile, fixed, sites (IFSs). IFSs occur ... More
The effect of cytosine arabinoside upon mitochondrial staining kinetics in human hematopoietic cells.
AuthorsHaanen C, Muus P, Pennings A
JournalHistochemistry
PubMed ID3487533
'The measurement of time correlated intracellular mitochondrial staining with 3,3''-dipentyloxacarbocyanine [Di-O-C5(3)] appeared of interest to define the optimal staining conditions. Mitochondrial staining of lymphocytes, monocytes and granulocytes results in different fluorescence signals, related to the numbers of mitochondria, that are present in the cells of these various cell types. Alterations ... More
Disruptive effects of tris and sodium lauroyl sarcosinate on the outer membrane of Pseudomonas cepacia shown by fluorescent probes.
AuthorsAnwar H, Brown MR, Britten AZ, Lambert PA
JournalJ Gen Microbiol
PubMed ID6195304
'The disruptive effects of Tris buffer and sodium lauroyl sarcosinate (Sarkosyl) on the outer membrane (OM) of Pseudomonas cepacia were investigated with several fluorescent probes. Tris increased the permeability of the OM to 6-anilino-l-naphthalenesulphonic acid and 2-p-toluidinylnaphthalene-6-sulphonate. The degree of damage to the OM was enhanced when the pH was ... More
Tumor necrosis factor as an activator of human granulocytes. Potentiation of the metabolisms triggered by the Ca2+-mobilizing agonists.
AuthorsYuo A, Kitagawa S, Suzuki I, Urabe A, Okabe T, Saito M, Takaku F
JournalJ Immunol
PubMed ID2537361
'TNF stimulated superoxide (O2-) release directly in human granulocytes in a dose-dependent manner (1 to 1000 U/ml), although its potency was weak. TNF-induced O2- release was inhibited by cAMP agonists or ionomycin, and was not accompanied with an increase in cytoplasmic free Ca2+ [( Ca2+]i) and membrane potential changes (depolarization). ... More
Measurement of transmembrane potentials in Rhodospirillum rubrum chromatophores with an oxacarbocyanine dye.
AuthorsPick U, Avron M
JournalBiochim Biophys Acta
PubMed ID820380
'The fluorescent dye 3,3-dipentyloxacarbocyanine (OCC) can be used as a fluorescence probe to measure transmembrane potentials across Rhodospirillum rubrum chromatophore membranes. A reversible fluorescence increase is observed in the light which is sensitive to inhibitors, permeable ions and uncouplers. Partial interchangeability between the electrical potential and the proton concentration gradient ... More
Reproducible high yield sequencing of proteins electrophoretically separated and transferred to an inert support.
AuthorsMoos M, Nguyen NY, Liu TY
JournalJ Biol Chem
PubMed ID3360771
'A method allowing initial sequencing yields of 60-85% to be consistently obtained from samples prepared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretic transfer is described in detail. Conducting electrophoresis at a pH near neutrality is the single most important of the modifications made to earlier procedures, but pre-electrophoresis in ... More
Neutrophil activation studied using the two indirect probes of membrane potential which respond by different fluorescence mechanisms.
AuthorsSeligmann B, Gallin JI
JournalAdv Exp Med Biol
PubMed ID6178272
Use of a fluorescent dye to measure drug-induced changes in the membrane potential of boar spermatozoa.
AuthorsPeterson RN, Bundman D, Freund M
JournalLife Sci
PubMed ID345031
Proceedings: The use of a fluorescent probe to demonstrate the interaction of calcium and oestradiol-17 beta in the thymic lymphocyte plasma membrane.
AuthorsMorgan JI, Bramhall JS, Britten AZ, Perris AD
JournalJ Endocrinol
PubMed ID1084908
The use of fluorescent probe to monitor alterations in trans-membrane potential in single cell suspensions.
AuthorsBramhall JS, Morgan JI, Perris AD, Britten AZ
JournalBiochem Biophys Res Commun
PubMed ID825119
Correlation between membrane potential changes and superoxide production in human granulocytes stimulated by phorbol myristate acetate. Evidence for defective activation in chronic granulomatous disease.
AuthorsWhitin JC, Chapman CE, Simons ER, Chovaniec ME, Cohen HJ
JournalJ Biol Chem
PubMed ID6243652
Dye indicators of membrane potential.
AuthorsWaggoner AS
JournalAnnu Rev Biophys Bioeng
PubMed ID383007
Potential-sensitive molecular probes in membranes of bioenergetic relevance.
AuthorsSmith JC
JournalBiochim Biophys Acta
PubMed ID2178682
Hydroperoxide-induced damage to alveolar macrophage function and membrane integrity: alterations in intracellular-free Ca2+ and membrane potential.
AuthorsForman HJ, Dorio RJ, Skelton DC
JournalArch Biochem Biophys
PubMed ID2827576
Cell membrane potential analysis.
AuthorsShapiro HM
JournalMethods Cell Biol
PubMed ID7532258
Adenosine affects the f-met-leu-phe induced changes in cytosolic Ca2+ and in membrane potential of human granulocytes.
AuthorsCapuozzo E, Crifò C, Siems W, Salerno C
JournalBiochem Mol Biol Int
PubMed ID8220253
Indo 1 acetoxymethyl ester and 3,3-dipentyloxacarbocyanine iodide have been used to investigate the effect of adenosine on the changes in cytosolic Ca2+ concentration and in membrane potential of human granulocytes activated by the chemotactic tripeptide f-met-leu-phe. Adenosine does not significantly affect negative (hyperpolarizing) potential changes as well as calcium accumulation ... More
Recent progress on structural interactions of the endoplasmic reticulum.
AuthorsTerasaki M
JournalCell Motil Cytoskeleton
PubMed ID2178783
Progress has been made recently in understanding certain aspects of the structure of the ER. It is very likely that kinesin, dynein, and myosin are associated with the ER and are responsible for distributing the fluid membranes of the ER by interaction of these motors with microtubules and actin filaments. ... More
Localization of endoplasmic reticulum in living and glutaraldehyde-fixed cells with fluorescent dyes.
AuthorsTerasaki M, Song J, Wong JR, Weiss MJ, Chen LB
JournalCell
PubMed ID6432338
Certain fluorescent dyes, previously reported to localize mitochondria, when used at higher concentrations also localize a continuous net-like structure in both living and glutaraldehyde-fixed cells. A similar reticular structure can be detected by phase-contrast microscopy and whole-mount electron microscopy in potassium permanganate-fixed cells as well. This structure is mostly tubular, ... More
Evaluation of murine lymphocyte membrane potential, intracellular free calcium, and interleukin-2 receptor expression upon exposure to 1,1-dimethylhydrazine.
AuthorsFrazier DE, Tarr MJ, Olsen RG
JournalToxicol Lett
PubMed ID1609436
The effects of 1,1-dimethylhydrazine (UDMH) on several early events associated with lymphocyte activation were examined. The concentration of intracellular calcium ([Ca2+]i) and membrane potential of murine lymphocytes were found to be altered upon exposure to UDMH; [Ca2+]i was increased in murine thymocytes, while splenocytes exhibited membrane hyperpolarization. In addition, interleukin-2 ... More
Lipid and salt effects on carbocyanine dye-induced photo-voltages in bilayer membranes.
Voltage transients induced by flash illumination of bilayer membranes with sorbed dye, 3,3' -dimethyl-2,2' -oxacarbocyanine chloride (diO-C1-3-CI), vary with membrane lipid composition and aqueous solution salt concentration. The voltage transients are probably induced by physical movements of sorbed dye molecules following photo-isomerization. Increased salt (NaCl and NaBr) concentrations in the ... More
Assessment of amphotericin B susceptibility in Leishmania infantum promastigotes by flow cytometric membrane potential assay.
AuthorsAzas N, Di Giorgio C, Delmas F, Gasquet M, Timon-David P
JournalCytometry
PubMed ID9181307
Flow cytometry was used for measuring the effects of amphotericin B on the membrane of Leishmania infantum strains. The technique was adapted from the rapid flow cytometric membrane potential assay developed by Ordonez and Wehman (Cytometry 22:154-157, 1995) for evaluating antibiotic-susceptibility of Candida species. The study consisted of measuring membrane ... More
Diverse subcellular locations of cryptogein-induced reactive oxygen species production in tobacco Bright Yellow-2 cells.
AuthorsAshtamker C, Kiss V, Sagi M, Davydov O, Fluhr R,
JournalPlant Physiol
PubMed ID17277088
Reactive oxygen species (ROS) play a crucial role in many cellular responses and signaling pathways, including the oxidative burst defense response to pathogens. We have examined very early events in cryptogein-induced ROS production in tobacco (Nicotiana tabacum) Bright Yellow-2 suspension cells. Using Amplex Red and Amplex Ultra Red reagents, which ... More
Monitoring of relative mitochondrial membrane potential in living cells by fluorescence microscopy.
AuthorsJohnson LV, Walsh ML, Bockus BJ, Chen LB
JournalJ Cell Biol
PubMed ID6783667
Permeant cationic fluorescent probes are shown to be selectively accumulated by the mitochondria of living cells. Mitochondria-specific interaction of such molecules is apparently dependent on the high trans-membrane potential (inside negative) maintained by functional mitochondria. Dissipation of the mitochondrial trans-membrane and potential by ionophores or inhibitors of electron transport eliminates ... More
Rapid flow cytometric antibiotic susceptibility assay for Staphylococcus aureus.
AuthorsOrdóñez JV, Wehman NM
JournalCytometry
PubMed ID8243210
Conventional techniques for antibiotic susceptibility testing usually require 24 h or more to produce accurate results. This long waiting period to demonstrate antibiotic action is necessary because such tests rely on growth (or the lack of it), when the microorganisms are incubated in the presence of the antibiotic. In an ... More
Proton permeability of sarcoplasmic reticulum vesicles.
AuthorsMeissner G, Young RC
JournalJ Biol Chem
PubMed ID7391050
The proton permeability of rabbit skeletal muscle sarcoplasmic reticulum vesicles was investigated by means of membrane potential measurements. Diffusion potentials were generated in sarcoplasmic reticulum vesicles, rat liver microsomes, and sarcoplasmic reticulum phospholipid vesicles by transferring vesicles from low to high pH. Potentials were measured using the voltage-sensitive fluorescent dye ... More
Amphotericin B susceptibility testing of Candida lusitaniae isolates by flow cytofluorometry: comparison with the Etest and the NCCLS broth macrodilution method.
AuthorsFavel A, Peyron F, De Méo M, Michel-Nguyen A, Carrière J, Chastin C, Regli P
JournalJ Antimicrob Chemother
PubMed ID11252328
A flow cytofluorometric susceptibility test (FCST) was used for rapid determination of the susceptibility of Candida lusitaniae isolates to amphotericin B. The test is based on the decrease in fluorescence intensity of cells stained with 3,3'-dipentyloxacarbocyanine iodide (DiOC5(3)), a membrane potential-sensitive cationic dye, after drug treatment. A total of 58 ... More
Evaluation of a flow cytofluorometric method for rapid determination of amphotericin B susceptibility of yeast isolates.
AuthorsPeyron F, Favel A, Guiraud-Dauriac H, El Mzibri M, Chastin C, Dumenil G, Regli P
JournalAntimicrob Agents Chemother
PubMed ID9210680
A rapid-flow cytofluorometric susceptibility test for in vitro amphotericin B testing of yeasts was evaluated and compared to the National Committee for Clinical Laboratory Standards (NCCLS) M27-T reference broth macrodilution method. The flow cytofluorometric method is based on the detection of decreased green fluorescence intensity of cells stained with DiOC5(3), ... More
Defective membrane potential changes in neutrophils from human neonates.
AuthorsSacchi F, Hill HR
JournalJ Exp Med
PubMed ID6481304
In an attempt to determine the mechanism of the profound defect in chemotaxis observed in the polymorphonuclear leukocytes (PMN) of human neonates, we have examined membrane potential changes and alterations in free intracellular calcium following chemotactic factor stimulation. Following exposure to formyl-methionyl-leucyl-phenylalanine (FMLP), PMN from adult donors (11) showed a ... More
Intracellular turnover of fluorescein diacetate. Influence of membrane ionic gradients on fluorescein efflux.
AuthorsProsperi E
JournalHistochem J
PubMed ID2387757
The influence of the membrane ionic gradient on the efflux of Fluorescein after intracellular turnover of Fluorescein diacetate was studied in HeLa cells. The kinetics of Fluorescein efflux was monitored by determining with flow cytometry the decrease in fluorescence intensity of single cells. Alterations of the Na+ and K+ gradients ... More
Membrane depolarization of human B cells follows stimulation by either anti-mu or B-cell growth factor, but only anti-mu causes cell volume changes.
AuthorsYokoyama WM, Chien MM, Engardt SE, Aguiar SW, Ashman RF
JournalHum Immunol
PubMed ID3131276
Human peripheral blood B cells were separated from monocytes and T cells, depleted of null cells by an anti-Leu 9 rosetting technique, and fractionated on discontinuous Percoll gradients to yield a highly purified, small, dense B-cell population. These cells responded to F(ab')2 goat anti-mu at 10 and 100 micrograms/ml with ... More
Optical probe responses on sarcoplasmic reticulum. Oxacarbocyanines.
AuthorsRussell JT, Beeler T, Martonosi A
JournalJ Biol Chem
PubMed ID154518
Absorbance and fluorescence changes of oxacarbocyanine dyes during ATP-induced Ca2+ transport in rabbit sarcoplasmic reticulum were analyzed. The response of the probes is complex and contains contributions from the binding of Ca2+ and ATP to the membrane. In a medium of 0.12 M KCl and 5 mM MgCl2, the fluorescence ... More
Mitochondria in cultured human muscle cells depleted of mitochondrial DNA.
AuthorsHerzberg NH, Middelkoop E, Adorf M, Dekker HL, Van Galen MJ, Van den Berg M, Bolhuis PA, Van den Bogert C
JournalEur J Cell Biol
PubMed ID8223726
Cultured human muscle cells were depleted of mitochondrial DNA (mtDNA) by prolonged treatment with ethidium bromide (EB). In these respiration-deficient muscle cells neither cytochrome c oxidase activity nor mtDNA were detectable. However, mitochondrial matrix enzymes remained present and were localized in mitochondria-like organelles, as shown by subcellular fractionation. Metabolic labeling ... More
B cell activation. I. Anti-immunoglobulin-induced receptor cross-linking results in a decrease in the plasma membrane potential of murine B lymphocytes.
AuthorsMonroe JG, Cambier JC
JournalJ Exp Med
PubMed ID6602204
We report analyses of the effect of anti-Fab antibodies on plasma membrane potential of mouse B lymphocytes. Results indicate that divalent fragments of anti-Fab antibodies mitogenic for B cells stimulate membrane depolarization detectable by cytofluorometric analysis of 3,3'-dipentyloxacarbocyanine iodide-stained cells. Depolarization is detectable within 5 min of exposure to ligand ... More
Ionic changes in the mitotic apparatus at the metaphase/anaphase transition.
AuthorsWolniak SM, Hepler PK, Jackson WT
JournalJ Cell Biol
PubMed ID6833373
We have employed a series of permeant, nontoxic, fluorescent probes to detect changes in ionic conditions within the mitotic apparatus of living endosperm cells of Haemanthus during the transition from metaphase to anaphase. Fluorescence emission intensity measurements from the spindle for chlorotetracycline (CTC) decline before the onset of anaphase, indicating ... More
Electroblotting onto activated glass. High efficiency preparation of proteins from analytical sodium dodecyl sulfate-polyacrylamide gels for direct sequence analysis.
AuthorsAebersold RH, Teplow DB, Hood LE, Kent SB
JournalJ Biol Chem
PubMed ID3949810
We have developed a new method for the isolation of proteins for microsequencing. It consists of electrophoretic transfer (electroblotting) of proteins or their cleavage fragments onto activated glass filter paper sheets immediately after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The proteins are immobilized on the glass fiber sheets by ... More
Protein-mediated exchange of synthetic phosphatidylcholines into synaptosomal membranes.
AuthorsNorth P, Fleischer S
JournalBiochim Biophys Acta
PubMed ID6712950
A phosphatidylcholine (PC) exchange protein from bovine liver was used to exchange endogenous synaptosomal membrane PC's with PC's of defined fatty-acid composition from phospholipid vesicles. Up to 50% of the total synaptosomal PC could be exchanged during a 3 h incubation with PC's which were in the liquid-crystalline state at ... More
Human T cells require IL-2 but not G1/S transition to acquire susceptibility to Fas-mediated apoptosis.
AuthorsFournel S, Genestier L, Robinet E, Flacher M, Revillard JP
JournalJ Immunol
PubMed ID8906804
The interaction between Fas ligand and Fas, both expressed on activated T cells, is the major pathway in the regulation of activation-induced cell death. However, activated T cells that express membrane Fas are initially resistant to anti-Fas-induced apoptosis and become susceptible only after proliferation in vitro. Since IL-2 is known ... More
The effects of phorbol myristate acetate and chemotactic peptide on transmembrane potentials and cytosolic free calcium in mature granulocytes evolve sequentially as the cells differentiate.
AuthorsSullivan R, Melnick DA, Malech HL, Meshulam T, Simons ER, Lazzari KG, Proto PJ, Gadenne AS, Leavitt JL, Griffin JD
JournalJ Biol Chem
PubMed ID3492493
We isolated myeloid precursors from human marrow and studied the effects of phorbol myristate acetate (PMA) and N-formyl-methionyl-leucyl-phenylalanine (fMLP) upon transmembrane potentials and cytosolic calcium ([Ca2+]i) as the cells matured. Using a panel of fluorescent probes, we found that membrane depolarization induced by PMA and fMLP in granulocytes, and elevation ... More
Lymphocyte membrane potential and Ca2+-sensitive potassium channels described by oxonol dye fluorescence measurements.
AuthorsWilson HA, Chused TM
JournalJ Cell Physiol
PubMed ID2413058
A method is described for quantitative measurement of lymphocyte transmembrane electrical potential difference (psi) by flow cytometric recording of the oxonol dye fluorescence of single cells. Both the simultaneous collection and analysis of multiple optical parameters and the use of a negatively charged oxonol probe allowed more accurate measurement of ... More
Polymorphonuclear leukocyte heterogeneity of Fc receptor expression and membrane potential in human neonates.
AuthorsMasuda K, Kinoshita Y, Hasui M, Kobayashi Y
JournalBiol Neonate
PubMed ID1797118
In order to clarify the relationship among polymorphonuclear neutrophil (PMN)-related phenomena, i.e. the Fc receptor expression, the ionic condition of the plasma membrane and chemotactic response, we determined percentage of EA rosette-forming PMN (RFN), membrane potential and PMN chemotaxis in 15 neonates. RFN was assessed by the whole blood method ... More
Rapid detection of efflux pumps and their relation with drug resistance in yeast cells.
AuthorsPrudêncio C, Sansonetty F, Sousa MJ, Côrte-Real M, Leão C
JournalCytometry
PubMed ID10655560
BACKGROUND: Cell drug resistance can be due to the presence of active efflux pumps (AEP). Identification of yeast cells with a resistance phenotype is important either from a clinical, agricultural or biotechnological point of view. Rapid and reliable methods to detect AEP can be therefore very useful. METHODS: Some yeast ... More
Effects of recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSFrh) on transmembrane electrical potentials in granulocytes: relationship between enhancement of ligand-mediated depolarization and augmentation of superoxide anion (O2-) production.
When human granulocytes that have been primed with recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSFrh) are activated by ligands that stimulate the respiratory burst, the amount of superoxide anion (O2-) they generate is significantly increased. We have found that the accelerated rate of O2- release occurring under these conditions is accompanied ... More
Stimulation of transient elevations in cytosolic Ca2+ is related to inhibition of Pi transport in OK cells.
AuthorsMiyauchi A, Dobre V, Rickmeyer M, Cole J, Forte L, Hruska KA
JournalAm J Physiol
PubMed ID2396674
Stimulation of changes in cytosolic free calcium by parathyroid hormone was determined in three opossum kidney (OK) cell types, OK wild-type, OKP clone, and OKH clone. All three types of OK cells express parathyroid hormone (PTH)-sensitive adenylate cyclase and adenosine 3',5'-cyclic monophosphate (cAMP) production. However, only the OK wild-type and ... More
Lipopolysaccharide priming of human neutrophils for an enhanced respiratory burst. Role of intracellular free calcium.
Lipopolysaccharide (LPS) pretreatment "primes" neutrophils to release increased amounts of superoxide anion (O2-) when stimulated. We investigated the molecular basis of this enhanced activity. Comparison of kinetic parameters of the respiratory burst NADPH oxidase in unstimulated LPS-primed and control neutrophils disclosed a similar Km for NADPH and no difference was ... More
Adaptation of human neutrophil responsiveness to the chemoattractant N-formylmethionylleucylphenylalanine. Heterogeneity and/or negative cooperative interaction of receptors.
AuthorsSeligmann BE, Fletcher MP, Gallin JI
JournalJ Biol Chem
PubMed ID6281264
The dose-response characteristics of the neutrophil 3-3'-dipentyloxacarbocyanine (di-O-C5(3)) fluorescence response to repetitive stimulation with the chemoattractant N-formylmethionylleucylphenylalanine (fMet-Leu-Phe) were studied. Neutrophils exposed to the chemoattractant fMet-Leu-Phe at less than 5 X 10(-8) M subsequently responded only to higher concentrations of fMet-Leu-Phe. This stimulus-induced modification of neutrophil responsiveness involved a reversible ... More
Effects of the myeloperoxidase system on fluorescent probes of granulocyte membrane potential.
AuthorsWhitin JC, Clark RA, Simons ER, Cohen HJ
JournalJ Biol Chem
PubMed ID6267057
Activation of normal or myeloperoxidase-deficient human granulocytes by phorbol myristate acetate resulted in an initial membrane depolarization as indicated by an increase in fluorescence of the lipophilic cation probe of membrane potential, 3,3'-dipropylthiodicarbocyanine. A subsequent apparent hyperpolarization (decrease in fluorescence) was observed in normal but not myeloperoxidase-deficient cells. Addition of ... More
Simultaneous measurements of cytosolic and mitochondrial Ca2+ transients in HT29 cells.
AuthorsRicken S, Leipziger J, Greger R, Nitschke R
JournalJ Biol Chem
PubMed ID9857027
Loading of HT29 cells with the Ca2+ dye fura-2/AM resulted in an nonhomogeneous intracellular distribution of the dye. Cellular compartments with high fura-2 concentrations were identified by correlation with mitochondrial markers, cellular autofluorescence induced by UV, and dynamic measurement of autofluorescence after inhibition of oxidative phosphorylation. Stimulation with carbachol (10(-4) ... More
Use of DiO-C5-3 to improve Hoechst 33342 uptake, resolution of DNA content, and survival of CHO cells.
AuthorsCrissman HA, Hofland MH, Stevenson AP, Wilder ME, Tobey RA
JournalExp Cell Res
PubMed ID2448153
Chinese hamster cells (line CHO) stained with either 9 microM Hoechst 33342 (HO) alone or in combination with the membrane potential fluorochrome DiO-C5-3 (DiO) were analyzed using uv laser powers between 25 and 500 mW and sorted for determination of survival by a colony formation assay. The combination of HO-DiO ... More
Interrelationships of polymorphonuclear neutrophil membrane-bound calcium, membrane potential, and chemiluminescence: studies in single living cells.
Stimulated neutrophils show ionic fluxes that may function as "transducers" between stimuli and effector functions. Using fluorescent probes, patterns of membrane-associated calcium (chlortetracycline, CTC) and membrane potential (3-3'-dipentyloxacarbocyanine, di-O-C5 (3)) in single living human neutrophils were observed with a fluorescence microscope fitted with an image intensifier and photometer. Fluorescence changes ... More
Extracellular ATP stimulates an amiloride-sensitive sodium influx in human lymphocytes.
AuthorsWiley JS, Jamieson GP, Mayger W, Cragoe EJ, Jopson M
JournalArch Biochem Biophys
PubMed ID2369117
Extracellular ATP has been shown to increase the Na+ permeability of human lymphocytes by 3 to 12-fold. The kinetics of this ATP-induced response were studied by measuring 22Na+ influx into chronic lymphocytic leukemic lymphocytes incubated in low-sodium media without divalent cations. ATP-stimulated uptake of 22Na-ions was linear over 4 min ... More
Lysophosphatidylcholine cell depolarization: increased membrane permeability for use in the determination of cell membrane potentials.
Current techniques for the determination of cellular membrane potentials based on the uptake of a radiolabeled lipophilic cation, [3H]triphenylmethylphosphonium, and the cyanine dye, DiOC5(3), were analyzed in terms of the proportions of these probes which are accumulated due to potential-dependent and potential-independent forces. Measurements were made of probe uptake in ... More
Simultaneous measurement of stimulus-induced changes in cytoplasmic Ca2+ and in membrane potential of human neutrophils.
AuthorsLazzari KG, Proto PJ, Simons ER
JournalJ Biol Chem
PubMed ID3755432
The activation of human neutrophils by chemotactic peptides evokes a rapid change in membrane potential and an increase in cytoplasmic Ca2+ levels. These events are followed up to a minute later by detectable levels of microbicidal agents formed by the oxidative burst. Except for the latter, the sequence of events ... More
Kinetics of transport of dialkyloxacarbocyanines in multidrug-resistant cell lines overexpressing P-glycoprotein: interrelationship of dye alkyl chain length, cellular flux, and drug resistance.
AuthorsWadkins RM, Houghton PJ
JournalBiochemistry
PubMed ID7893683
The membrane transport properties of a series of dialkyloxacarbocyanine [DiOCn(3)] dyes in multidrug-resistant KB cell lines were investigated to determine the influence of alkyl chain length on the ability of p-glycoprotein (i) to protect cells from the toxicity of the dyes and (ii) to affect the plasma membrane flux of ... More
Fast responses of bacterial membranes to virus adsorption: a fluorescence study.
AuthorsBayer ME, Bayer MH
JournalProc Natl Acad Sci U S A
PubMed ID7029540
After collision with their host cells, virus particles may remain mobile on cell surfaces until they become attached at firm binding sites. We propose that a virion will arrive within a typical median time at such a site, generating a membrane signal such as an increased membrane fluorescence in cells ... More
Graded responses of human neutrophils induced by serum-treated zymosan.
AuthorsWhitin JC, Ryan DH, Cohen HJ
JournalBlood
PubMed ID2996662
A modified zymosan preparation was used to probe the interaction of particulate stimuli with human neutrophils (PMNs). After extraction with alkali and detergent, the zymosan particles retained their ability to be opsonized in serum and to stimulate PMNs. Serum-treated zymosan (STZ) induced dose-dependent superoxide (O2-) production and membrane potential depolarization ... More
B cell activation. VI. Effects of exogenous diglyceride and modulators of phospholipid metabolism suggest a central role for diacylglycerol generation in transmembrane signaling by mIg.
AuthorsCoggeshall KM, Cambier JC
JournalJ Immunol
PubMed ID2981089
Previous evidence indicates that in vitro activators of protein kinase C, such as phorbol myristate acetate (PMA), are able to induce early activation events in murine B cells, including membrane depolarization and increased I-A antigen expression. These same events are induced by specific antigen and anti-receptor antibody. This evidence suggests ... More
Neutrophil hyperpolarization in response to a chemotactic peptide.
AuthorsLazzari KG, Proto P, Simons ER
JournalJ Biol Chem
PubMed ID2162827
The chemotactic peptide formylmethionyl-leucyl-phenylalanine (fMLP), at concentrations below 10(-9) M, elicits a sustained increase in the human neutrophil's membrane potential within 10 s of its addition. This hyperpolarization, detected with the fluorescent cationic potentiometric probes, 3,3'-dipentyloxacarbocyanine (diO-C5-(3)), and 1,1'-dipropyl-3,3,3',3'-tetramethylindocarbocyanine iodide (diI-C3-(3)), and with the anionic probe bis-(1,3-diethylthiobarbituric)trimethine oxonol (bis-oxonol), is ... More
Monitoring human neutrophil granule secretion by flow cytometry: secretion and membrane potential changes assessed by light scatter and a fluorescent probe of membrane potential.
AuthorsFletcher MP, Seligmann BE
JournalJ Leukoc Biol
PubMed ID3855949
Purified human peripheral blood polymorphonuclear neutrophils (PMN) were incubated at 37 degrees C with the fluorescent membrane potential sensitive cyanine dye di-O-C(5)(3) and exposed to a number of stimulatory agents (N-formylmethionylleucylphenylalanine (FMLP), cytochalasin B (cyto B) + FMLP, phorbol myristate acetate (PMA). Flow cytometry was utilized to measure changes in ... More
Inhibition by linoleic acid hydroperoxide of alveolar macrophage superoxide production: effects upon mitochondrial and plasma membrane potentials.
AuthorsForman HJ, Kim E
JournalArch Biochem Biophys
PubMed ID2552924
Linoleic acid hydroperoxide (LOOH) is a naturally occurring product of lipid peroxidation. Incubation of rat alveolar macrophages with LOOH produced alterations of membrane properties and function at concentrations of LOOH as low as 0.1 microM. These included phorbol myristate acetate (PMA)-stimulated superoxide production, mitochondrial membrane potential, and plasma membrane potentials. ... More
ERM (ezrin/radixin/moesin)-based molecular mechanism of microvillar breakdown at an early stage of apoptosis.
AuthorsKondo T, Takeuchi K, Doi Y, Yonemura S, Nagata S, Tsukita S
JournalJ Cell Biol
PubMed ID9348291
Breakdown of microvilli is a common early event in various types of apoptosis, but its molecular mechanism and implications remain unclear. ERM (ezrin/radixin/moesin) proteins are ubiquitously expressed microvillar proteins that are activated in the cytoplasm, translocate to the plasma membrane, and function as general actin filament/plasma membrane cross-linkers to form ... More
Inhibition of mitochondrial and Paracoccus denitrificans NADH-ubiquinone reductase by oxacarbocyanine dyes. A structure-activity study.
AuthorsAnderson WM, Wood JM, Anderson AC
JournalBiochem Pharmacol
PubMed ID8512593
In this study, we determined that three structurally related oxacarbocyanine dyes, 3,3'-diethyloxacarbocyanine (DiOC2(3)), 3,3'-dipentyloxacarbocyanine (DiOC5(3)), and 3,3'-dihexyloxacarbocyanine (DiOC6(3)), and one oxadicarbocyanine, 3,3'-diethyloxadicarbocyanine (DiOC2(4)), inhibit bovine heart mitochondrial NADH oxidase activity and one of them, DiOC6(3), inhibits Paracoccus denitrificans NADH oxidase activity. The mitochondrial I50 values were 9 microM (DiOC2(3)), approximately ... More
Flow cytometric analysis of rat striatal nerve terminals.
AuthorsWolf ME, Kapatos G
JournalJ Neurosci
PubMed ID2563283
Methods were developed for the analysis and isolation of striatal nerve terminals (synaptosomes) using fluorescence-activated cell sorting (FACS). Comparison of the light-scattering properties of synaptosomal and mitochondrial fractions indicated that particles in the synaptosomal fraction were generally larger and more sensitive to hypotonic lysis, consistent with results obtained by other ... More
Use of lipophilic probes of membrane potential to assess human neutrophil activation. Abnormality in chronic granulomatous disease.
AuthorsSeligmann BE, Gallin JI
JournalJ Clin Invest
PubMed ID6249851
Previous studies using membrane potential sensitive probes have provided evidence that chemotactic factors elicit membrane potential changes in normal human neutrophils (PMN). In addition to stimulation of PMN motility, chemotactic factors also stimulate degranulation and superoxide ion (O-2) generation and it has been suggested that alteration of membrane potential activates ... More
PMN heterogeneity: long-term stability of fluorescent membrane potential responses to the chemoattractant N-formyl-methionyl-leucyl-phenylalanine in healthy adults and correlation with respiratory burst activity.
AuthorsFletcher MP, Seligmann BE
JournalBlood
PubMed ID3017475
Polymorphonuclear neutrophils (PMNs) were isolated from 24 healthy adults 20 to 61 years of age and the proportion of cells that demonstrated depolarization responses to the synthetic chemotaxin N-formyl-methionyl-leucyl-phenylalanine (FMLP) were enumerated using the lipophilic fluorescent cyanine dye 3,3'-di-pentyl-oxacarbocyanine [di-O-C(5)(3)] and flow cytometry. The membrane potential responses were correlated to ... More
Prostaglandin E1 inhibits N-formyl-methionyl-leucyl-phenylalanine-mediated depolarization responses by decreasing the proportion of responsive cells without affecting chemotaxin-induced forward light scatter changes.
AuthorsFletcher MP
JournalJ Immunol
PubMed ID3693900
Hypaque-Ficoll-purified human polymorphonuclear neutrophils (PMN) equilibrated with the membrane potential-sensitive probe 3,3'dipentyloxacarbocyanine [di-O-C(5)(3)] were incubated with buffer or cytochalasin B (cyto B) followed by incubation with prostaglandin E1 (PGE1) (0 to 10(-5) M) for 5 min at 37 degrees C. The cells were then stimulated with N-formyl-methionyl-leucyl-phenylalanine (FMLP) (0 to ... More
Lymphocyte membrane potential assessed with fluorescent probes.
AuthorsRink TJ, Montecucco C, Hesketh TR, Tsien RY
JournalBiochim Biophys Acta
PubMed ID6153065
The membrane potential of mouse spleen lymphocytes has been assessed with two fluorescent probes. 3,3'-Dipropylthiadicarbocyanine (diS-C3-(5)) was used for most of the experiments. Solutions with high K+ concentrations depolarised the cells. Valinomycin, an inophore which adds a highly K+-selective permeability membranes, slightly hyperpolarised cells in standard (6 mM K+) solution, ... More
Slow fluorescent indicators of membrane potential: a survey of different approaches to probe response analysis.
AuthorsPlásek J, Sigler K
JournalJ Photochem Photobiol B
PubMed ID8691353
Basic tenets related to the use of three main classes of potentiometric redistribution fluorescent dyes (carbocyanines, oxonols, and rhodamines) are discussed in detail. They include the structure/function relationship, formation of nonfluorescent (H-type) and fluorescent (J-type) dimers and higher aggregates, probe partitioning between membranes and medium and binding to membranes and ... More
Stimulation by leukotriene D4 of increases in the cytosolic concentration of calcium in dimethylsulfoxide-differentiated HL-60 cells.
AuthorsBaud L, Goetzl EJ, Koo CH
JournalJ Clin Invest
PubMed ID3477571
The C6-sulfidopeptide leukotrienes C4 (LTC4) and D4 (LTD4) evoked increases in the cytosolic concentration of intracellular calcium ([Ca+2]i) in dimethylsulfoxide-differentiated HL-60 cells, as assessed by the fluorescence of quin-2. The increases in [Ca+2]i reached a peak within 15-90 s, attained 50% of the maximum level at 1.2 nM LTD4 and ... More
Neutrophil defect associated with malignant infantile osteopetrosis.
AuthorsBeard CJ, Key L, Newburger PE, Ezekowitz RA, Arceci R, Miller B, Proto P, Ryan T, Anast C, Simons ER
JournalJ Lab Clin Med
PubMed ID3021878
We studied the phagocytes from three infants with malignant osteopetrosis and from their families in an attempt to define further the phagocyte abnormalities associated with this disorder. The rapid membrane potential depolarization in response to the soluble stimuli formylmethionylleucylphenylalanine (FMLP) and phorbol myristate acetate (PMA) served as a measure of ... More
Phorbol ester-stimulated human neutrophil membrane depolarization is dependent on Ca2(+)-regulated Cl- efflux.
AuthorsMyers JB, Cantiello HF, Schwartz JH, Tauber AI
JournalAm J Physiol
PubMed ID2171344
The ionic basis of phorbol 12-myristate 13-acetate (PMA)-stimulated membrane depolarization in the human neutrophil has not previously been established. Alterations in cation permeability are probably not directly responsible for the depolarization response, since the rate or Rb+ influx or efflux is unchanged upon PMA stimulation, and although Na+ fluxes are ... More
Interaction of chemotactic factors with human polymorphonuclear leukocytes: studies using a membrane potential-sensitive cyanine dye.
AuthorsSeligmann BE, Gallin EK, Martin DL, Shain W, Gallin JI
JournalJ Membr Biol
PubMed ID6770097
Changes in the fluorescence intensity of the dye 3-3' dipentyloxacarbocyanine were measured in suspensions of purified human peripheral blood polymorphonuclear leukocytes (PMNs) during exposure to the chemotactic factors N-formyl-methionyl-leucyl-phenylalanine (f-met-leu-phe) and partially purified C5a. Incubation of PMNs with dye resulted in a stable fluorescence reflecting the resting membrane potential of ... More
Comparison of indirect probes of membrane potential utilized in studies of human neutrophils.
AuthorsSeligmann BE, Gallin JI
JournalJ Cell Physiol
PubMed ID6302103
Four indirect probes of membrane potential, triphenylmethylphosphonium ion (TPMP+), 3,3'dipentyloxacarbocyanine [di-O-C5(3)], 3,3'dipentylindocarbocyanine [di-I-C5(3)], and 3,3'dipropylthiodicarbocyanine [di-S-C3(5)] have been used to study neutrophil (PMN) activation. The data extend previous studies indicating that the cyanine dye di-S-C3(5) not only exhibits a different fluorescence response mechanism from di-O-C5(3) [and di-I-C5(3)] but also that ... More
Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analyses.
AuthorsDavey HM, Kell DB
JournalMicrobiol Rev
PubMed ID8987359
The most fundamental questions such as whether a cell is alive, in the sense of being able to divide or to form a colony, may sometimes be very hard to answer, since even axenic microbial cultures are extremely heterogeneous. Analyses that seek to correlate such things as viability, which is ... More
Lipid bodies and lipid body formation in an oleaginous fungus, Mortierella ramanniana var. angulispora.
AuthorsKamisaka Y, Noda N, Sakai T, Kawasaki K
JournalBiochim Biophys Acta
PubMed ID10320801
Mortierella ramanniana var. angulispora accumulates triacylglycerol (TG) in lipid bodies. Studies on lipid transport into lipid bodies are essential for elucidating mechanisms of lipid body formation. We used fluorescent dyes and fluorescent lipid analogs to visualize lipid body formation with a confocal laser scanning microscope. Different sizes of lipid bodies ... More
Phorbol myristate acetate potentiates superoxide release and membrane depolarization without affecting an increase in cytoplasmic free calcium in human granulocytes stimulated by the chemotactic peptide, lectins and the calcium ionophore.
AuthorsOhsaka A, Saito M, Suzuki I, Miura Y, Takaku F, Kitagawa S
JournalBiochim Biophys Acta
PubMed ID2835980
We investigated the inter-relationships of superoxide (O2-) release, membrane depolarization and an increase in cytoplasmic free Ca2+, [Ca2+]i, in human granulocytes stimulated by various agonists. When concanavalin A or the Ca2+ ionophore ionomycin was used as stimulus, an increase in [Ca2+]i clearly preceded the onset of membrane depolarization, which was ... More
Voltage-sensitive cyanine dye fluorescence signals in lymphocytes: plasma membrane and mitochondrial components.
AuthorsWilson HA, Seligmann BE, Chused TM
JournalJ Cell Physiol
PubMed ID2413057
The origin of the cyanine dye fluorescence signal in murine and human peripheral blood leukocytes was investigated using the oxa- and indo-carbocyanines di-O-C5(3) and di-I-C5(3). Fluorescence signals from individual cells suspended with nanomolar concentrations of the dyes were measured in a flow cytometer modified to permit simultaneous four-parameter analysis (including ... More
Merocyanine 540 as a fluorescent probe of altered membrane phospholipid asymmetry in activated whole blood platelets.
AuthorsWatala C, Waczulikova I, Wieclawska B, Rozalski M, Gresner P, Gwozdzinski K, Mateasik A, Sikurova L
JournalCytometry
PubMed ID12442312
BACKGROUND: Platelet activation leads to the loss of a natural asymmetry of membrane phospholipids (PL) and the subsequent exposure of negatively charged PL in platelets with procoagulant activity that can be monitored routinely with annexin V (AN-V). METHODS: Flow cytometric analysis of merocyanine 540 (MC540) binding may be the alternate ... More
Granulocytes without degranulation: neutrophil function in granule-depleted cytoplasts.
AuthorsKorchak HM, Roos D, Giedd KN, Wynkoop EM, Vienne K, Rutherford LE, Buyon JP, Rich AM, Weissmann G
JournalProc Natl Acad Sci U S A
PubMed ID6308664
Neutrophils respond to a variety of stimuli by generating superoxide anion, degranulating, and aggregating. Because it has been suggested that fusion of granules with the plasmalemma (degranulation) is necessary for aggregation and superoxide anion generation, we have tested whether these responses can be demonstrated in "neutrophilic cytoplasts" (granule-free vesicles of ... More
Liposome-cell interactions in vitro: effect of liposome surface charge on the binding and endocytosis of conventional and sterically stabilized liposomes.
AuthorsMiller CR, Bondurant B, McLean SD, McGovern KA, O'Brien DF
JournalBiochemistry
PubMed ID9737866
The cellular uptake of liposomes is generally believed to be mediated by adsorption of liposomes onto the cell surface and subsequent endocytosis. This report examines the effect of liposome surface charge on liposomal binding and endocytosis in two different cell lines: a human ovarian carcinoma cell line (HeLa) and a ... More