Low-dose estrogen therapy ameliorates experimental autoimmune encephalomyelitis in two different inbred mouse strains.
AuthorsBebo BF Jr,Fyfe-Johnson A,Adlard K,Beam AG,Vandenbark AA,Offner H
JournalJournal of immunology (Baltimore, Md. : 1950)
PubMed ID11160259
Spatial variations in growth rate within Klebsiella pneumoniae colonies and biofilm.
AuthorsWentland EJ,Stewart PS,Huang CT,McFeters GA
JournalBiotechnology progress
PubMed ID8652119
The use of acridine orange to visualize and quantify spatial variations in growth rate within Klebsiella pneumoniae colonies and biofilm was investigated. Bacterial colonies supported on polycarbonate filter membranes were grown on R2A agar plates. Some colonies were sampled for cell enumeration, while others were cryoembedded, sectioned, and stained with ... More
Rapid assessment of physiological status in Escherichia coli using fluorescent probes.
AuthorsPorter J, Edwards C, Pickup RW
JournalJ Appl Bacteriol
PubMed ID7592133
'Rapid and direct viability assessment of Escherichia coli in filtered, sterile lake water was possible using multiparameter flow cytometry. Fluorescent dyes were used as probes for different cellular functions (membrane potential, membrane integrity and intracellular enzyme activity), which were correlated with the ability of the cells to respond to nutrient ... More
Defective acidification of intracellular organelles in cystic fibrosis.
AuthorsBarasch J, Kiss B, Prince A, Saiman L, Gruenert D, al-Awqati Q
JournalNature
PubMed ID1712081
'The phenotype of cystic fibrosis (CF) includes abnormalities in transepithelial transport of Cl- (refs 1-5), decreased sialylation and increased sulphation and fucosylation of glycoproteins, and lung colonization with Pseudomonas. It is not apparent how these abnormalities are interrelated, nor how they result from loss of function of the CF gene-encoded ... More
Rapid movements of vimentin on microtubule tracks: kinesin-dependent assembly of intermediate filament networks.
AuthorsPrahlad V, Yoon M, Moir RD, Vale RD, Goldman RD
JournalJ Cell Biol
PubMed ID9763428
'The assembly and maintenance of an extended intermediate filament (IF) network in fibroblasts requires microtubule (MT) integrity. Using a green fluorescent protein-vimentin construct, and spreading BHK-21 cells as a model system to study IF-MT interactions, we have discovered a novel mechanism involved in the assembly of the vimentin IF cytoskeleton. ... More
Two-photon fluorescence absorption and emission spectra of dyes relevant for cell imaging.
AuthorsBestvater F, Spiess E, Stobrawa G, Hacker M, Feurer T, Porwol T, Berchner-Pfannschmidt U, Wotzlaw C, Acker H
JournalJ Microsc
PubMed ID12423261
'Two-photon absorption and emission spectra for fluorophores relevant in cell imaging were measured using a 45 fs Ti:sapphire laser, a continuously tuneable optical parametric amplifier for the excitation range 580-1150 nm and an optical multichannel analyser. The measurements included DNA stains, fluorescent dyes coupled to antibodies as well as organelle ... More
Simple epithelium keratins 8 and 18 provide resistance to Fas-mediated apoptosis. The protection occurs through a receptor-targeting modulation.
AuthorsGilbert S, Loranger A, Daigle N, Marceau N
JournalJ Cell Biol
PubMed ID11514590
'Keratins 8 and 18 belong to the keratin family of intermediate filament (IF) proteins and constitute a hallmark for all simple epithelia, including the liver. Hepatocyte IFs are made solely of keratins 8 and 18 (K8/K18). In these cells, the loss of one partner via a targeted null mutation in ... More
Intra-endosomal pH-sensitive recruitment of the Arf-nucleotide exchange factor ARNO and Arf6 from cytoplasm to proximal tubule endosomes.
AuthorsMaranda B, Brown D, Bourgoin S, Casanova JE, Vinay P, Ausiello DA, Marshansky V
JournalJ Biol Chem
PubMed ID11278939
'Kidney proximal tubule epithelial cells have an extensive apical endocytotic apparatus that is critical for the reabsorption and degradation of proteins that traverse the glomerular filtration barrier and that is also involved in the extensive recycling of functionally important apical plasma membrane transporters. We show here that an Arf-nucleotide exchange ... More
Nerve growth factor signaling regulates motility and docking of axonal mitochondria.
AuthorsChada SR, Hollenbeck PJ
JournalCurr Biol
PubMed ID15268858
'Axonal transport is thought to distribute mitochondria to regions of the neuron where their functions are required. In cultured neurons, mitochondrial transport responds to growth cone activity, and this involves both a transition between motile and stationary states of mitochondria and modulation of their anterograde transport activity. Although the exact ... More
Chronic hypoxia protects against gamma-irradiation-induced apoptosis by inducing bcl-2 up-regulation and inhibiting mitochondrial translocation and conformational change of bax protein.
AuthorsCuisnier O, Serduc R, Lavieille JP, Longuet M, Reyt E, Riva C
JournalInt J Oncol
PubMed ID12963983
'Malignant tumours contain zones of chronic or acute hypoxia, which influence their prognosis and progression. The goal of our study was to understand the role of hypoxia in radio-resistance in a squamous cell carcinoma cell line of the head and neck (KB-3-1 cells). Cell growth was evaluated by Trypan blue ... More
Some fluorescent counterstains for neuroanatomical studies.
AuthorsSchmued LC, Swanson LW, Sawchenko PE
JournalJ Histochem Cytochem
PubMed ID6174560
'Methods for counterstaining neural tissue that contains fluorescent markers have been developed. Acridine orange is useful for localizing cells that are retrogradely labelled with the fluorescent tracers true blue, bisbenzimide, and nuclear yellow because at low concentrations it yields a green Nissl stain when excited with blue, but not with ... More
Prelysosomal acidic vacuoles in Dictyostelium discoideum.
AuthorsPadh H, Lavasa M, Steck TL
JournalJ Cell Biol
PubMed ID2466039
'We have examined the ameba Dictyostelium discoideum for evidence of a discrete, prelysosomal, acidic receiving compartment in endocytosis. We observed in the cytoplasm abundant round vacuoles with diameters up to 2 microns that concentrated acridine orange by a process inhibited by 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl). They were therefore taken to be acidic. ... More
Aminoacridines, potent inhibitors of protein kinase C.
AuthorsHannun YA, Bell RM
JournalJ Biol Chem
PubMed ID3258596
'Acridine orange, acridine yellow G, and related compounds potently inhibited protein kinase C (Ca2+/phospholipid-dependent enzyme) activity and phorbol dibutyrate binding. Inhibition was investigated in vitro using Triton X-100 mixed micellar assays (Hannun, Y. A., Loomis, C. R., and Bell, R. M. (1985) J. Biol. Chem. 260, 10039-10043 and Hannun, Y. ... More
Patch-clamp detection of macromolecular translocation along nuclear pores.
AuthorsBustamante JO, Varanda WA
JournalBraz J Med Biol Res
PubMed ID9698781
'The present paper reviews the application of patch-clamp principles to the detection and measurement of macromolecular translocation along the nuclear pores. We demonstrate that the tight-seal ''gigaseal'' between the pipette tip and the nuclear membrane is possible in the presence of fully operational nuclear pores. We show that the ability ... More
Close proximity of Cys64 and Cys140 in the delta subunit of Escherichia coli F1-ATPase.
AuthorsZiegler M, Xiao R, Penefsky HS
JournalJ Biol Chem
PubMed ID8307987
'The delta subunit of the F1-ATPase from Escherichia coli contains 2 cysteine residues, one at position 64 and the second at position 140 of the amino acid sequence. These residues were specifically labeled with sulfhydryl reagents in this study without labeling other -SH groups in the enzyme. Modification of Cys140 ... More
Use of fluorescent dyes for measurement and localization of organelles associated with Ca2+ store release in human neutrophils.
AuthorsDavies EV, Blanchfield H, Hallett MB
JournalCell Biol Int
PubMed ID9693835
'Fura-2 and its lipid analogue, FFP-18, were used to measure changes in cytosolic free Ca2+ concentration within human neutrophils. Whereas fura-2 was employed to monitor cytosolic Ca2+ increases throughout the cytosol, FFP-18 was used to monitor Ca2+ changes only near the membrane. This latter probe was incorporated into the plasma ... More
Cell-cell interactions in the process of differentiation of thyroid epithelial cells into follicles: a study by microinjection and fluorescence microscopy on in vitro reconstituted thyroid follicles.
AuthorsMunari-Silem Y, Mesnil M, Selmi S, Bernier-Valentin F, Rabilloud R, Rousset B
JournalJ Cell Physiol
PubMed ID2125602
'Thyroid cells, cultured in the presence of thyroid stimulating hormone, reorganized within 36-48 hr into follicular structures, the in vitro reconstituted thyroid follicles or RTF. By microinjection of fluorescent probes either into the neoformed intrafollicular lumen (IL) or into cells forming the follicles, we have studied the development and some ... More
Inhibition of the bovine-heart mitochondrial F1-ATPase by cationic dyes and amphipathic peptides.
AuthorsBullough DA, Ceccarelli EA, Roise D, Allison WS
JournalBiochim Biophys Acta
PubMed ID2527062
'The bovine heart mitochondrial F1-ATPase is inhibited by a number of amphiphilic cations. The order of effectiveness of non-peptidyl inhibitors examined as assessed by the concentration estimated to produce 50% inhibition (I0.5) of the enzyme at pH 8.0 is: dequalinium (8 microM), rhodamine 6G (10 microM), malachite green (14 microM), ... More
Ca2+ oscillations induced by hormonal stimulation of individual fura-2-loaded hepatocytes.
AuthorsKawanishi T, Blank LM, Harootunian AT, Smith MT, Tsien RY
JournalJ Biol Chem
PubMed ID2473983
'Isolated rat hepatocytes were loaded with the Ca2+ indicator fura-2 to measure cytosolic free Ca2+ concentrations ([Ca2+]i) in individual cells by digital ratio imaging microscopy. Stimulation with 0.1 nM vasopressin, 0.5 microM phenylephrine, or 0.5 microM ATP caused repetitive spikes of high [Ca2+]i in a high percentage of cells, in ... More
Apoptosis in shed human corneal cells.
AuthorsEstil S, Primo EJ, Wilson G
JournalInvest Ophthalmol Vis Sci
PubMed ID11006225
'PURPOSE: To determine whether shear forces applied to the corneal epithelium by the repeated insertion and removal of a hydrogel contact lens alter the size and number of cells removed and to determine the contribution of apoptosis to this process. METHODS; Human corneal cells were collected from eight healthy subjects ... More
Fc receptor off-signal in the B cell involves apoptosis.
AuthorsAshman RF, Peckham D, Stunz LL
JournalJ Immunol
PubMed ID8683155
'By linking surface Ig to the FcR Fc gamma RII on the mouse B lymphocyte surface, whole anti-Ig has been shown to block cell cycle entry and subsequent Ab production, a phenomenon called the "Fc receptor off-signal." IL-4 or blocking Ab to Fc gamma RII, present with whole anti-Ig, restores ... More
Energy-dependent accumulation of daunorubicin into subcellular compartments of human leukemia cells and cytoplasts.
AuthorsSlapak CA, Lecerf JM, Daniel JC, Levy SB
JournalJ Biol Chem
PubMed ID1350280
'Anthracycline accumulation was evaluated by flow cytometry or radiolabeled drug assays in cells and cytoplasts (enucleated cells) prepared from parental and multidrug-resistant human K562 leukemia cells. Treatment with energy inhibitors, such as dinitrophenol (DNP) or sodium azide/deoxyglucose, led to a marked decrease in daunorubicin accumulation in parental cells and cytoplasts. ... More
Further characterisation of the in situ terminal deoxynucleotidyl transferase (TdT) assay for the flow cytometric analysis of apoptosis in drug resistant and drug sensitive leukaemic cells.
AuthorsChapman RS, Chresta CM, Herberg AA, Beere HM, Heer S, Whetton AD, Hickman JA, Dive C
JournalCytometry
PubMed ID7587710
'Apoptosis, originally defined by specific morphological changes, is characterised biochemically by non-random cleavage of DNA. Depending on cell type, this DNA cleavage proceeds from 300 and 50kbp fragments prior to, concomitantly with, or in the absence of 180bp integer fragmentation. Incorporation into fragmented DNA of biotin-labelled nucleotides by terminal deoxynucleotidyl ... More
Hg2+ and Cu+ are ionophores, mediating Cl-/OH- exchange in liposomes and rabbit renal brush border membranes.
AuthorsKarniski LP
JournalJ Biol Chem
PubMed ID1527045
'Organometals, including organomercurials, are capable of mediating Cl-/OH- exchange across lipid membranes by forming neutral ion pairs. In this study, the ability of inorganic metals to catalyze Cl-/OH- exchange was examined. In the presence of an inwardly directed chloride gradient, HgCl2 at concentrations as low as 30 nM resulted in ... More
Comparison of two flow cytometric assays for cellular RNA--acridine orange and propidium iodide.
AuthorsWallen CA, Higashikubo R, Dethlefsen LA
JournalCytometry
PubMed ID6184207
'Two flow cytometric assays for cellular RNA, two-step acridine orange (TSAO) and propidium iodide (PI), were compared with each other and with ultraviolet (uv) spectrophotometry of RNA to determine their ability to quantitate cellular RNA and to differentiate between proliferating and quiescent (Q) cells. The model system used for these ... More
CpG oligodeoxyribonucleotides rescue mature spleen B cells from spontaneous apoptosis and promote cell cycle entry.
AuthorsYi AK, Chang M, Peckham DW, Krieg AM, Ashman RF
JournalJ Immunol
PubMed ID9637502
'Isolated murine splenic B cells undergo spontaneous apoptosis. Motifs containing unmethylated CpG dinucleotides in bacterial DNA or in synthetic oligodeoxynucleotides (ODN) are known to activate murine B cells. Now we show that ODN that induce spleen B cell cycle entry also inhibit spontaneous apoptosis in a sequence-specific fashion. Reversal of ... More
A rapid method for measuring apoptosis and dual-color immunofluorescence by single laser flow cytometry.
AuthorsSchmid I, Uittenbogaart CH, Keld B, Giorgi JV
JournalJ Immunol Methods
PubMed ID8157993
'A sensitive method for quantification of cells undergoing apoptosis that permits the simultaneous measurement of dual-color cell surface immunofluorescence is presented. Unfixed cells are stained with 7-amino-actinomycin D (7-AAD) for discrimination of live from early apoptotic cells and from cells which have lost membrane integrity (late apoptotic or necrotic, dead ... More
Determination of intracellular organelles implicated in daunorubicin cytoplasmic sequestration in multidrug-resistant MCF-7 cells using fluorescence microscopy image analysis.
AuthorsBour-Dill C, Gramain MP, Merlin JL, Marchal S, Guillemin F
JournalCytometry
PubMed ID10655559
'BACKGROUND: Anthracycline resistance is known to be mediated by P-glycoprotein (P-gp) or multidrug-resistance related protein (MRP) as well as intracellular sequestration of drugs. METHODS: The resistance phenotype of doxorubicin-selected MCF-7(DXR) human breast adenocarcinoma cell line was characterized by cellular and nuclear daunorubicin efflux, P-gp and MRP expression and apoptosis induction. ... More
The lipid fluidity of rat colonic brush-border membrane vesicles modulates Na+-H+ exchange and osmotic water permeability.
AuthorsBrasitus TA, Dudeja PK, Worman HJ, Foster ES
JournalBiochim Biophys Acta
PubMed ID3002472
'Brush-border membrane vesicles were prepared from rat colonic epithelial cells. Steady-state fluorescence polarization techniques, using the fluorophores 1,6-diphenyl-1,3,5-hexatriene and DL-12-(9-anthroyl)stearic acid (12-AS), revealed that benzyl alcohol (25-75 mM) but not methyl alcohol (50-125 mM) significantly increased the fluidity of these vesicles. Benzyl alcohol (50 and 75 mM) but not methyl ... More
A cell-free system for regulated exocytosis in PC12 cells.
AuthorsAvery J, Ellis DJ, Lang T, Holroyd P, Riedel D, Henderson RM, Edwardson JM, Jahn R
JournalJ Cell Biol
PubMed ID10648564
'We have developed a cell-free system for regulated exocytosis in the PC12 neuroendocrine cell line. Secretory vesicles were preloaded with acridine orange in intact cells, and the cells were sonicated to produce flat, carrier-supported plasma membrane patches with attached vesicles. Exocytosis resulted in the release of acridine orange which was ... More
Rapid analysis of normal and abnormal cell types in human semen and testis biopsies by flow cytometry.
AuthorsEvenson DP, Melamed MR
JournalJ Histochem Cytochem
PubMed ID6186729
'A simple, rapid procedure is described that quantitates RNA content and DNA content/chromatin condensation for each of many possible cell types and differentiation levels of the cells present in human semen. A fresh semen sample (1-6 hr postemission) or frozen sample (allowing samples to be accumulated and sent to a ... More
Transmembrane molecular pump activity of Niemann-Pick C1 protein.
AuthorsDavies JP, Chen FW, Ioannou YA
JournalScience
PubMed ID11125140
'Niemann-Pick C1 (NPC1) disease is characterized by cholesterol accumulation in lysosomes and aberrant feedback regulation of cellular cholesterol homeostasis. We provide evidence that the NPC1 protein has homology with the resistance-nodulation-division (RND) family of prokaryotic permeases and may normally function as a transmembrane efflux pump. Studies of acriflavine loading in ... More
Synthesis of Bcl-2 in response to anthracycline treatment may contribute to an apoptosis-resistant phenotype in leukemic cell lines.
AuthorsDurrieu F, Belaud-Rotureau MA, Lacombe F, Dumain P, Reiffers J, Boisseau MR, Bernard P, Belloc F
JournalCytometry
PubMed ID10554162
'BACKGROUND: Some forms of chemoresistance in leukemia may start from failure of tumour cells to successfully undergo apoptosis and Bcl-2 may play a role in this defect. Therefore, we evaluated the Bcl-2 content and synthesis in relation with the apoptotic potential in leukemic cell lines after anthracycline treatment. METHODS: U937, ... More
Cellular basis for the response to second-order motion cues in Y retinal ganglion cells.
AuthorsDemb JB, Zaghloul K, Sterling P
JournalNeuron
PubMed ID11719210
'We perceive motion when presented with spatiotemporal changes in contrast (second-order cue). This requires linear signals to be rectified and then summed in temporal order to compute direction. Although both operations have been attributed to cortex, rectification might occur in retina, prior to the ganglion cell. Here we show that ... More
YOPRO-1 permits cytofluorometric analysis of programmed cell death (apoptosis) without interfering with cell viability.
AuthorsIdziorek T, Estaquier J, De Bels F, Ameisen JC
JournalJ Immunol Methods
PubMed ID7561136
'In the absence of cell permeabilization, the impermeant nuclear dye YOPRO-1 permits accurate analysis of apoptosis using cytofluorometry or fluorescent microscopy. Several immune cell populations were studied including dexamethasone-treated thymocytes, irradiated peripheral blood mononuclear cells and a growth factor-depleted tumor B cell line. Excellent correlation values were found with acridine ... More
Presence of a plant-like proton-pumping pyrophosphatase in acidocalcisomes of Trypanosoma cruzi.
AuthorsScott DA, de Souza W, Benchimol M, Zhong L, Lu HG, Moreno SN, Docampo R
JournalJ Biol Chem
PubMed ID9705361
'The vacuolar-type proton-translocating pyrophosphatase (V-H+-PPase) is an enzyme previously described in detail only in plants. This paper demonstrates its presence in the trypanosomatid Trypanosoma cruzi. Pyrophosphate promoted organellar acidification in permeabilized amastigotes, epimastigotes, and trypomastigotes of T. cruzi. This activity was stimulated by K+ ions and was inhibited by Na+ ... More
[The effect of biologically active compounds on lysosome fusion with phagosomes and the F-actin content in mouse peritoneal macrophages and on the status of the lysosomal membranes in mouse hepatocytes]
'Effects of biologically active compounds bilirubin (BR, 0.1 and 0.2 mM), chelerythrine (CR, 0.1 and 0.5 mM) and farmorubicin (FR, 0.6 and 6.0 mM) on the phagosome-lysosome fusion (P-LF) were studied using fluorescent dye acridine orange for lysosomal labelling and yeast cells as a target. To investigate mechanisms of these ... More
Cell death-associated translocation of plasma membrane components induced by CTL.
AuthorsKawasaki Y, Saito T, Shirota-Someya Y, Ikegami Y, Komano H, Lee MH, Froelich CJ, Shinohara N, Takayama H
JournalJ Immunol
PubMed ID10779768
'In the very early stages of target cell apoptosis induced by CTL, we found that fluorescence of labeling probes of the target plasma membrane, such as N-(3-triethylammoniumpropyl)-4-(p-dibutylaminostyryl)pyridin ium dibromide (FM1-43), was translocated into intracellular membrane structures including nuclear envelope and mitochondria. This translocation was associated with the execution of CTL-mediated ... More
Polyanionic agents do not inhibit phagosome-lysosome fusion in cultured macrophages.
AuthorsGoren MB, Vatter AE, Fiscus J
JournalJ Leukoc Biol
PubMed ID3468192
'The survival of some intracellular pathogens within macrophages may be aided by an ability of the organism to antagonize, from within the entrapping phagosome, its fusion with lysosomes. On the other hand, certain polyanionic agents have been implicated in imposing a similar block to fusion from the lysosomal domain--because the ... More
Mapping mechanisms and charting the time course of premature cell senescence and apoptosis: lysosomal dysfunction and ganglioside accumulation in endothelial cells.
AuthorsPatschan S, Chen J, Gealekman O, Krupincza K, Wang M, Shu L, Shayman JA, Goligorsky MS,
JournalAm J Physiol Renal Physiol
PubMed ID17928415
'Endothelial cells subjected to glycated collagen I develop premature senescence within 3-5 days, as revealed by increased senescence-associated beta-galactosidase activity, decreased proliferation, and an increase in cell size. Here, we analyzed the time course and possible mechanisms of this process. Lysosomal integrity studies revealed a rapid collapse of pH gradient ... More
Evidence for uptake of vital dye by activated rat peritoneal mast cells: an in vitro imaging study.
AuthorsWei JY, Go VL, Taché Y, Kruger L
JournalNeuroimage
PubMed ID9343581
'Uptake of material from surrounding medium by activated rat peritoneal mast cells (PMCs) was studied using in vitro peritoneal eluate cells, the vital fluorescent dye sulforhodamine B (SFRM-B), secretagogue compound 48/80, and an imaging technique. PMCs, which undergo different states of degranulation, are shown to possess the ability to take ... More
Hydrogen peroxide inhibits the vacuolar H+-ATPase in brain synaptic vesicles at micromolar concentrations.
AuthorsWang Y, Floor E
JournalJ Neurochem
PubMed ID9453558
'Hydrogen peroxide (H2O2) is produced from several sources in brain and may be involved in neurodegeneration and second messenger signaling. Little is known about the effects of H2O2 on transmitter storage in brain synaptic vesicles. Neurotransmitter uptake into synaptic vesicles is driven by an electrochemical proton gradient generated by the ... More
Flow cytometric detection of erythropoietic cytotoxicity in mouse bone marrow.
AuthorsBenning VM, Maratrat MB, Fournier EC, Melcion CP, Cordier AC
JournalJ Histochem Cytochem
PubMed ID1701186
'Erythroblast proliferation and maturation in bone marrow are the processes leading to the formation of polychromatic erythrocytes (PE) and normochromatic erythrocytes (NE), respectively. PE contain RNA but no DNA, and can therefore be distinguished both from NE (which lack both RNA and DNA) and from nucleated cells (which contain both ... More
DNA distribution analysis of type II pneumocytes by laser flow cytometry: technical considerations.
AuthorsUhal BD, Rannels DE
JournalAm J Physiol
PubMed ID1928365
'Optimal conditions were established for determination of cell cycle phase fractions of freshly isolated or cultured adult rat type II pneumocytes (T2P). Propidium iodide staining of ethanol-fixed cells treated with ribonuclease (RNase) consistently yielded histograms with low coefficients of variation. Contaminating cells and cell clumps were eliminated during data acquisition ... More
Discrepancy in assessment of phagosome-lysosome fusion with two lysosomal markers in murine macrophages infected with Candida albicans.
AuthorsMor N, Goren MB
JournalInfect Immun
PubMed ID3298062
'Phagosome-lysosome fusion (P-LF) was studied in cultured mouse resident peritoneal macrophages after phagocytosis of Candida albicans. The macrophages were labeled with acridine orange (AO), the electronopaque colloidal Thorotrast, or both markers. After phagocytosis of heat-killed C. albicans, both markers were delivered to more than 95% of phagosomes. After ingestion of ... More
Lysosomal destabilization in p53-induced apoptosis.
AuthorsYuan XM, Li W, Dalen H, Lotem J, Kama R, Sachs L, Brunk UT
JournalProc Natl Acad Sci U S A
PubMed ID11959917
'The tumor suppressor wild-type p53 can induce apoptosis. M1-t-p53 myeloid leukemic cells have a temperature-sensitive p53 protein that changes its conformation to wild-type p53 after transfer from 37 degrees C to 32 degrees C. We have now found that these cells showed an early lysosomal rupture after transfer to 32 ... More
Increase in acridine orange (AO) fluorescence intensity of monocytes cultured in plastic tissue culture plates as measured by flow cytometry.
AuthorsMyc A, Pizzolo JG, Dygulski K, Melamed MR
JournalCytometry
PubMed ID1547653
'Although the green-red fluorescence of AO is an accepted measure of DNA-RNA content, respectively, it is actually a measure of the fluorescence of dye bound to nucleic acids, and may vary with changes in accessibility to the dye. It has been shown for example that extraction of nuclear proteins results ... More
Association of cytoplasmic free Ca2+ gradients with subcellular organelles.
AuthorsWahl M, Sleight RG, Gruenstein E
JournalJ Cell Physiol
PubMed ID1537888
'Previous investigations have identified gradients of intracellular free (Ca2+)i (Ca2+i) in the cytoplasm of human fibroblasts. In this study we have compared the spatial distribution of these gradients with the subcellular distribution of cytoplasmic organelles. Using the Ca(2+)-sensitive dye fura-2 and organelle-specific fluorescent dyes, we have found that the highest ... More
A non-exchangeable fluorescent phospholipid analog as a membrane traffic marker of the endocytic pathway.
AuthorsWillem J, ter Beest M, Scherphof G, Hoekstra D
JournalEur J Cell Biol
PubMed ID2076704
'The fluorescent phospholipid analog N-(lissamine rhodamine B sulfonyl)phosphatidylethanolamine (N-Rh-PE) was inserted into the plasma membrane of Baby hamster kidney cells at low temperature (2 degrees C). The mobility characteristics of the analog--as revealed by fluorescence photobleaching recovery--were very similar to those of membrane-inserted 1-acyl-2[6-[N-(7-nitro-2,1,3-benzoxadiazol-4-yl) amino]caproyl] phosphatidylcholine (C6-NBD-PC). Upon warming to ... More
The vacuolar H+-ATPase of clathrin-coated vesicles is reversibly inhibited by S-nitrosoglutathione.
AuthorsForgac M
JournalJ Biol Chem
PubMed ID9880499
'It has been previously demonstrated that the vacuolar H+-ATPase (V-ATPase) of clathrin-coated vesicles is reversibly inhibited by disulfide bond formation between conserved cysteine residues at the catalytic site on the A subunit (Feng, Y., and Forgac, M. (1994) J. Biol. Chem. 269, 13224-13230). Proton transport and ATPase activity of the ... More
In vitro testing of the potential for orthopedic bone cements to cause apoptosis of osteoblast-like cells.
AuthorsCiapetti G, Granchi D, Savarino L, Cenni E, Magrini E, Baldini N, Giunti A
JournalBiomaterials
PubMed ID11761181
'The purpose of this study was to investigate in vitro the apoptosis- and/or necrosis-inducing potential of polymethylmethacrylate (PMMA)-based bone cements for prosthetic surgery. Four bone cements widely used in orthopedics were tested as extracts onto osteoblast-like MG-63 cells and for comparison, HL-60 cells, which are remarkably sensitive to apoptotic stimuli. ... More
Development of resting membrane potentials of embryonic murine spinal cord cells evaluated by flow cytometric analysis.
AuthorsKrieger C, Puil E, Kim SU
JournalDev Neurosci
PubMed ID1647295
'The membrane electrical properties of embryonic murine spinal cord cells of embryonic day-10 (E10) to E14 were studied using a voltage-sensitive oxonol dye combined with fluorescence-activated cell sorting techniques. This period of development corresponds to the time when neurons differentiate from their neuroblast precursors. Bovine oligodendrocytes were also investigated in ... More
Exocytosis regulates urinary acidification in turtle bladder by rapid insertion of H+ pumps into the luminal membrane.
AuthorsGluck S, Cannon C, Al-Awqati Q
JournalProc Natl Acad Sci U S A
PubMed ID6289300
'Urinary acidification by the turtle bladder is due to a H+-ATPase that is located in the luminal membrane. The rate of H+ transport is stimulated by an increase in the ambient CO2. Using the fluorescent dye acridine orange, we showed that the mitochondria-rich cell of this equilibrium contains vesicles whose ... More
Interactions between pairs of DNA-binding dyes: results and implications of chromosome analysis.
AuthorsLatt SA, Sahar E, Eisenhard ME, Juergens LA
JournalCytometry
PubMed ID6168450
'A number of DNA-binding dyes, with spectral properties making them suitable as components of energy donor-acceptor pairs, are described. If such pairs are used to stain metaphase chromosomes, and if the energy acceptor (e.g., actinomycin D or methyl green) has a binding specificity opposite to the binding or fluorescence specificity ... More
Polyanionic agents as inhibitors of phagosome-lysosome fusion in cultured macrophages: evolution of an alternative interpretation.
AuthorsGoren MB, Vatter AE, Fiscus J
JournalJ Leukoc Biol
PubMed ID3468191
'Various natural and synthetic substances classified as polyanionics have been implicated in antagonizing phagosome-lysosome fusion in cultured macrophages. The phenomenon has been judged by comparing the transfer of selected markers from secondary lysosomes to phagosomes in control and in "polyanion" cells. Our earlier studies showed that use of one of ... More
Enzyme kinetic reactions and fluorochrome uptake rates measured in individual cells by laser scanning cytometry.
AuthorsBedner E, Melamed MR, Darzynkiewicz Z
JournalCytometry
PubMed ID9725553
'This study was designed to explore the utility of a microscope-based cytofluorometer, the laser scanning cytometer (LSC), for time-resolved kinetic measurements. This instrument measures fluorescence of individual cells rapidly, with high sensitivity and accuracy. Also recorded in a list mode fashion are the spatial X-Y coordinates of the cell on ... More
Sphingosine-induced apoptosis is dependent on lysosomal proteases.
AuthorsKågedal K, Zhao M, Svensson I, Brunk UT
JournalBiochem J
PubMed ID11583579
'We propose a new mechanism for sphingosine-induced apoptosis, involving relocation of lysosomal hydrolases to the cytosol. Owing to its lysosomotropic properties, sphingosine, which is also a detergent, especially when protonated, accumulates by proton trapping within the acidic vacuolar apparatus, where most of its action as a detergent would be exerted. ... More
Calpain activation in apoptosis.
AuthorsSquìer MK, Miller AC, Malkinson AM, Cohen JJ
JournalJ Cell Physiol
PubMed ID8163563
'Programmed cell death is an active process wherein the cell initiates a sequence of events culminating in the fragmentation of its DNA, nuclear collapse, and disintegration of the cell into small, membrane-bound apoptotic bodies. Examination of the death program in various models has shown common themes, including a rise in ... More
Depletion of intracellular Ca2+ by caffeine and ryanodine induces apoptosis of chinese hamster ovary cells transfected with ryanodine receptor.
AuthorsPan Z, Damron D, Nieminen AL, Bhat MB, Ma J
JournalJ Biol Chem
PubMed ID10764805
'Recent studies have suggested a central role for Ca(2+) in the signaling pathway of apoptosis and certain anti-apoptotic effects of Bcl-2 family of proteins have been attributed to changes in intracellular Ca(2+) homeostasis. Here we report that depletion of Ca(2+) from endoplasmic reticulum (ER) leads to apoptosis in Chinese hamster ... More
Interactions of nucleic acids with fluorescent dyes: spectral properties of condensed complexes.
AuthorsKapuscinski J
JournalJ Histochem Cytochem
PubMed ID1696951
'Interaction of cations with nucleic acids (NA) often results in condensation of the product. The driving force of aromatic cation-induced condensation is the cooperative interaction between ligand and single-stranded (ss) NA. This type of reaction is highly specific with regard to the primary and secondary structure of NA, and results ... More
Inositol trisphosphate and cyclic ADP-ribose-mediated release of Ca2+ from single isolated pancreatic zymogen granules.
'In pancreatic acinar cells low (physiological) agonist concentrations evoke cytosolic Ca2+ spikes specifically in the apical secretory pole that contains a high density of secretory (zymogen) granules (ZGs). Inositol 1,4,5-trisphosphate (IP3) is believed to release Ca2+ from the endoplasmic reticulum, but we have now tested whether the Ca(2+)-releasing messengers IP3 ... More
Optimisation of flow cytometric measurement of parasitaemia in plasmodium-infected mice.
AuthorsBarkan D, Ginsburg H, Golenser J
JournalInt J Parasitol
PubMed ID10779580
'Mouse malaria is often used as a model for drug testing. The results of drug trials are monitored by tedious (and consequently, sometimes inaccurate) microscopic counting of blood smears, or by flow cytometry. We suggest an improved, accurate and time-saving flow cytometric method for determination of parasitaemias in mice infected ... More
Analysis of early apoptotic events in individual cells by fluorescence intensity and polarization measurements.
AuthorsZurgil N, Shafran Y, Fixler D, Deutsch M
JournalBiochem Biophys Res Commun
PubMed ID11820802
'Apoptosis is a dynamic process of variable duration. The ability to continuously detect the death process occurring in single or subgroups of cells is therefore very important in identifying apoptotic cells within a complex population. The Individual Cell Scanner (ICS), a multiparametric, multilaser-based scanning static cytometer, was used in the ... More
An improved fluorochrome microassay for the detection of living and non-living intracellular bacteria in human neutrophils.
AuthorsHorn W, Hansmann C, Federlin K
JournalJ Immunol Methods
PubMed ID2414364
'Acridine orange fluorescence may be used to distinguish living from non-living intracellular bacteria in individual glass-adherent neutrophil granulocytes (PMN). An improvement of the original assay (Smith and Rommel, 1977; Pantazis and Kniker, 1979) is described which allows differentiation between ingested and cell-adherent bacteria. It is shown that this differentiation is ... More
Denaturation of nucleic acids induced by intercalating agents. Biochemical and biophysical properties of acridine orange-DNA complexes.
AuthorsKapuscinski J, Darzynkiewicz Z
JournalJ Biomol Struct Dyn
PubMed ID6400830
'At high binding densities acridine orange (AO) forms complexes with ds DNA which are insoluble in aqueous media. These complexes are characterized by high red- and minimal green-luminescence, 1:1 (dye/P) stoichiometry and resemble complexes of AO with ss nucleic acids. Formation of these complexes can be conveniently monitored by light ... More
Thallium mediates a rapid chloride/hydroxyl ion exchange through myelin lipid bilayers.
AuthorsDíaz RS, Monreal J
JournalMol Pharmacol
PubMed ID7808444
'We have investigated the effects of several heavy metal cations on the proton and chloride permeabilities of liposomes prepared with endogenous lipids from brain myelin, by monitoring the fluorescence emitted by acridine orange and N-(6-methoxyquinolyl)acetoethyl ester. In addition to Hg2+ and Cu+, nanomolar concentrations of Tl3+, but not Tl+, were ... More
Blockade of the apoptotic machinery by cyclosporin A redirects cell death toward necrosis in arterial endothelial cells: regulation by reactive oxygen species and cathepsin D.
AuthorsRaymond MA, Mollica L, Vigneault N, Désormeaux A, Chan JS, Filep JG, Hébert MJ
JournalFASEB J
PubMed ID12514115
'Blockade of the mitochondrial permeability transition pore (mPTP) by cyclosporin A (CsA) inhibits apoptosis in various cell types. However, use of CsA in humans is associated with damage to the arterial endothelium. We evaluated whether inhibition of the apoptotic machinery by CsA promotes other forms of cell death in arterial ... More
Denaturation and condensation of intracellular nucleic acids monitored by fluorescence depolarization of intercalating dyes in individual cells.
AuthorsBeisker W, Eisert WG
JournalJ Histochem Cytochem
PubMed ID2478614
'The intercalating binding of planar aromatic dye molecules to nucleic acids can be analyzed using fluorescence depolarization measurements of the dye molecules excited by linearly polarized light. In this study, we investigated the conformational changes of the intracellular DNA-dye complex in single cells. Flow cytometry, combined with a newly developed ... More
Decreased vacuolar acidification capacity in drug-resistant rat liver preneoplastic nodules.
AuthorsAndersson GN, Torndal UB, Eriksson LC
JournalCancer Res
PubMed ID2525416
'Rat liver nodules produced by intermittent 2-acetylaminofluorene feeding exhibit alterations in cell surface receptors reminiscent of impairment of vacuolar acidification. In this report, vacuolar acidification activity, measured as the ATP-dependent quenching of acridine orange, was characterized in liver and nodular membrane fractions using various ion-transport inhibitors and with respect to ... More
The lysosomal compartment as intracellular calcium store in MDCK cells: a possible involvement in InsP3-mediated Ca2+ release.
AuthorsHaller T, Dietl P, Deetjen P, Völkl H
JournalCell Calcium
PubMed ID8689673
'To test for a possible role of lysosomes in intracellular Ca2+ homeostasis, the effects of glycyl-L-phenylalanine-beta-naphthylamide (GPN), known to permeabilize these organelles by osmotic swelling, were studied in single MDCK cells. Fluorescence of acridine orange, rhodol green dextran, lysotracker green and FITC-dextran indicated that GPN (0.2 mmol/l) elicited a reversible ... More
Identification of F-reticulocytes by two-stage fluorescence image cytometry.
AuthorsOsterhout ML, Ohene-Frempong K, Horiuchi K
JournalJ Histochem Cytochem
PubMed ID8601699
'Precise determination of reticulocytes (young red blood cells) containing fetal hemoglobin (Hb F), F-reticulocytes, is important for assessment of the efficacy of drugs such as hydroxyurea and butyrate in elevating the levels of Hb F in patients with sickle cell disease (SCD) and beta-thalassemia. We developed a reliable and easily ... More
Proton translocating ATPase in lysosomal membrane ghosts. Evidence that alkaline Mg2+-ATPase acts as a proton pump.
AuthorsMoriyama Y, Takano T, Ohkuma S
JournalJ Biochem (Tokyo)
PubMed ID6146604
'Membrane ghosts were prepared from purified lysosomes (tritosomes) of rat liver by hypo-osmotic treatment. Mg2+-ATP-driven acidification was observed in the membrane ghosts using acridine orange as a fluorescent probe of the transmembrane pH gradient (delta pH). Its properties were the same as those of intact lysosomes reported previously (Ohkuma, S., ... More
Multiple spectral parameter imaging.
AuthorsWaggoner A, DeBiasio R, Conrad P, Bright GR, Ernst L, Ryan K, Nederlof M, Taylor D
JournalMethods Cell Biol
PubMed ID2648118
Identification of hypoxic and oxygenated tumor subpopulations with enzyme and fluorescent probes.
AuthorsFreitas I, Pontiggia P
JournalJ Photochem Photobiol B
PubMed ID2107296
Comparison of four DNA staining fluorescence dyes for measuring cell proliferation of lymphokine-activated killer (LAK) cells.
AuthorsVollenweider I, Groscurth P
JournalJ Immunol Methods
PubMed ID1374777
Different quiescence states of three culture cell lines detected by acridine orange staining of cellular RNA.
AuthorsBaisch H
JournalCytometry
PubMed ID2456898
'Three cell lines (CHO, L-929, and R1H) were investigated for their growth kinetics and the difference of exponential and quiescent state of monolayers in medium with and without serum (L-929). The noncycling populations of L-929 and R1H in medium with serum contained increased G1-phase percentages but also considerable proportions of ... More
Microchip-based capillary electrochromatography using packed beds.
AuthorsJemere AB, Oleschuk RD, Harrison DJ
JournalElectrophoresis
PubMed ID12973805
'Integration of a packed column onto a microchip for performance of capillary electrochromatography (CEC) is described. The quartz device incorporated a cross-injector, and a double weir trapping design for formation of 1, 2 and 5 mm long CEC columns. Three fluorescent dyes were baseline-resolved with plate numbers of 330,(330,000 plates/m; ... More
Oxygen radicals mediate cell inactivation by acridine dyes, fluorescein, and lucifer yellow CH.
AuthorsMartin JP, Logsdon N
JournalPhotochem Photobiol
PubMed ID3039547
'Acridine dyes, fluorescein and lucifer yellow CH are fluorescent photosensitizers used experimentally to selectively stain and photodynamically destroy eukaryotic cells and subcellular structures. We have determined that the mechanism of light- and oxygen-dependent inactivation of E. coli by these dyes involves oxygen radicals and hydrogen peroxide. All of the dyes ... More
Guanosine nucleotides modulate the inhibitory effect of brefeldin A on protein secretion.
AuthorsZeuzem S, Zimmermann P, Schulz I
JournalFEBS Lett
PubMed ID1908795
'Brefeldin A (BFA) causes rapid redistribution of Golgi proteins into the endoplasmic reticulum (ER), leaving no definable Golgi-apparatus, and blocks transport of proteins from the ER to distal secretory compartments of the cell. Using pulse-chase experiments the present study shows that BFA (1 microgram/ml) inhibits basal and CCK-stimulated protein secretion ... More
Inhibition of an oligomycin-sensitive ATPase by cationic dyes, some of which are atypical uncouplers of intact mitochondria.
AuthorsMai MS, Allison WS
JournalArch Biochem Biophys
PubMed ID6188413
'The inhibition of an oligomycin sensitive ATPase prepared from bovine heart submitochondrial particles (J.A. Berden and M.M. Voorn-Brouwer, 1978, Biochim. Biophys. Acta 501, 424-439) by a number of cationic dyes has been compared in order to develop a structure-function relationship. Two generalizations emerge from this comparison. First, the most effective ... More
The use of fluorescein diacetate and ethidium bromide as a viability stain for isolated islets of Langerhans.
AuthorsGray DW, Morris PJ
JournalStain Technol
PubMed ID2448922
'There is a need for a simple, rapid, sensitive method for assessing the viability of isolated islets of Langerhans. In this study the fluorescent dyes fluorescein diacetate (FDA) and ethidium bromide (EB) have been used to provide a viability assay for isolated rat islets. Discrimination of living from dead islets ... More
Crystalline ribosomes are present in brains from senile humans.
AuthorsO'Brien L, Shelley K, Towfighi J, McPherson A
JournalProc Natl Acad Sci U S A
PubMed ID6929549
'Paracrystalline inclusions known as Hirano bodies characteristically appear in the hippocampal region of the brains of humans exhibiting senile and presenile dementias as well as several other neurodegenerative diseases. We present evidence that the currently accepted model for those structures based on alternating filament sheets is not correct, but that ... More
SYBR Green I-induced fluorescence in cultured immune cells: a comparison with Acridine Orange.
AuthorsBriggs C, Jones M
JournalActa Histochem
PubMed ID16139877
'Using fluorescence microscopy, we explored the ability of cultured immune cells to take up aqueous SYBR Green I (SGI). SYBR Green I, a highly sensitive fluorescent nucleic acid stain, which preferentially binds to dsDNA over ssDNA or RNA with little background fluorescence from unbound molecules. A time course study over ... More
Differential trafficking of live and dead Mycobacterium marinum organisms in macrophages.
AuthorsBarker LP, George KM, Falkow S, Small PL
JournalInfect Immun
PubMed ID9119492
'We characterized the Mycobacterium marinum phagosome by using a variety of endocytic markers to follow the path of the bacteria through a mouse macrophage cell line. Using a laser confocal microscope, we found that the majority of viable M. marinum cells were in nonacidic vacuoles that did not colocalize with ... More
Macromolecule entrapment in human placental microvillous membrane vesicles.
AuthorsIllsley NP, Goodman JG, Verkman AS, Davis B
JournalAnal Biochem
PubMed ID2469349
'An osmotic lysis technique was developed to induce transient permeability in human placental microvillous membrane vesicles. The degree of vesicle opening and resealing was quantitated using the fluorescent markers, 6-carboxyfluorescein and fluorescein dextran. Compared to freeze-thaw and sonication methods, hypotonic lysis was significantly more efficient, causing greater than 90% lysis ... More
Prodigiosins as a new group of H+/Cl- symporters that uncouple proton translocators.
AuthorsSato T, Konno H, Tanaka Y, Kataoka T, Nagai K, Wasserman HH, Ohkuma S
JournalJ Biol Chem
PubMed ID9705273
'We reported previously (Kataoka, T., Muroi, M., Ohkuma, S., Waritani, T., Magae, J., Takatsuki, A., Kondo, S., Yamasaki, M., and Nagai, K. (1995) FEBS Lett. 359, 53-59) that prodigiosin 25-C uncoupled vacuolar H+-ATPase, inhibited vacuolar acidification, and affected glycoprotein processing. In the present study we show that prodigiosins (prodigiosin, metacycloprodigiosin, ... More
Tracking single secretory granules in live chromaffin cells by evanescent-field fluorescence microscopy.
AuthorsSteyer JA, Almers W
JournalBiophys J
PubMed ID10096921
'We have observed secretory granules beneath the plasma membrane of chromaffin cells. Using evanescent-field excitation by epiillumination, we have illuminated a thin layer of cytosol where cells adhere to glass coverslips. Up to 600 frames could be recorded at diffraction-limited resolution without appreciable photodynamic damage. We localized single granules with ... More
Photo-oxidative disruption of lysosomal membranes causes apoptosis of cultured human fibroblasts.
AuthorsBrunk UT, Dalen H, Roberg K, Hellquist HB
JournalFree Radic Biol Med
PubMed ID9215807
'Acridine orange (AO) is a lysosomotropic weak base, a metachromatic fluorochrome, and a photosensitizer, as well. Living cells that are exposed for a short period of time to this compound at low concentration, and under ordinary culture conditions, accumulate the drug within their acidic vacuolar compartment, giving rise to a ... More
Rapid identification and detection of parasitized human red cells by automated flow cytometry.
AuthorsWhaun JM, Rittershaus C, Ip SH
JournalCytometry
PubMed ID6354641
'Rapid and reliable identification of various human red cells parasites is important in many chemotherapeutic and immunologic studies. Because manual microscopic counting is tedious and imprecise, we have developed a simple diagnostic procedure for the automated flow cytometric detection of in vitro infected red cells, using a nucleic acid-binding fluorescent ... More
The involvement of protein phosphatases in the activation of ICE/CED-3 protease, intracellular acidification, DNA digestion, and apoptosis.
AuthorsMorana SJ, Wolf CM, Li J, Reynolds JE, Brown MK, Eastman A
JournalJ Biol Chem
PubMed ID8663484
'Many events in apoptosis have been identified but their temporal relationships remain obscure. Apoptosis in human ML-1 cells induced by etoposide is characterized by intracellular acidification, enhanced Hoechst 33342 fluorescence, DNA digestion, chromatin condensation, and proteolysis of poly(ADP-ribose) polymerase. This proteolysis is a marker for the action of ICE/CED-3 proteases, ... More
Cellular F-actin levels as a marker for cellular transformation: correlation with bladder cancer risk.
AuthorsRao JY, Hemstreet GP, Hurst RE, Bonner RB, Min KW, Jones PL
JournalCancer Res
PubMed ID2032215
'Previous findings in cultured cells that differentiated cells had markedly higher F-actin levels than undifferentiated cells (Cancer Res., 50: 2215-2220, 1990) suggested that quantitative F-actin measurements in urinary cells might provide diagnostic or prognostic information by identifying those individuals with cells tending towards a lower degree of differentiation. The feasibility ... More
ZnT-2, a mammalian protein that confers resistance to zinc by facilitating vesicular sequestration.
AuthorsPalmiter RD, Cole TB, Findley SD
JournalEMBO J
PubMed ID8617223
'A cDNA encoding a second zinc transporter (ZnT-2) was isolated from a rat kidney cDNA expression library by complementation of a zinc-sensitive BHK cell line. The protein predicted from the open reading frame of ZnT-2 cDNA has 359 amino acids and initiates with a CTG codon. It resembles ZnT-1 (a ... More
Assessment of islet cell viability using fluorescent dyes.
AuthorsBank HL
JournalDiabetologia
PubMed ID3322907
'A rapid fluorometric method has been developed to evaluate the viability of isolated islet cells. The assay differentiates between viable and nonviable cells by the simultaneous use of the inclusion and exclusion dyes acridine orange and propidium iodide. When viewed by fluorescent microscopy, viable cells fluoresce green, while nonviable cells ... More
Cell cycle-specific effects of tumor necrosis factor.
AuthorsDarzynkiewicz Z, Williamson B, Carswell EA, Old LJ
JournalCancer Res
PubMed ID6690064
'The immediate effect of tumor necrosis factor (TNF) added to cultures of L-cells is cytostasis, manifested as cell arrest in G2. This effect prevails during the initial 4 hr when the number of G2 cells increases markedly in the absence of any significant cell death in the culture. Shortly thereafter, ... More
Apoptotic cell nuclei favour aggregation and fluorescence quenching of DNA dyes.
AuthorsErenpreisa J, Freivalds T, Roach H, Alston R
JournalHistochem Cell Biol
PubMed ID9377226
'Apoptotic cell nuclei are known to stain hyperchromatically with absorption dyes and dimly with many DNA fluorochromes. We hypothesised that both optical phenomena have the same cause--the ability of apoptotic chromatin to aggregate cationic dyes. This hypothesis was tested using prednisolone-primed rat thymus, which is known to contain apoptotic cells. ... More
Fluorescence assay to monitor phagocytosis by bloodclot derived polymorphonuclear leucocytes. 2. Study of patients with psoriasis and the effect of exposing leucocytes to psoriatic skin scales.
AuthorsMusclow CE, Farkas-Himsley H, Kanee-Schachter R, Rubino K, Der S
JournalCytobios
PubMed ID2055104
'Comparisons of phagocytic parameters were carried out by a recently developed fluorescence test which is reproducible, simple and fast. Phagocytosis by polymorphonuclear leucocytes (PMNs) obtained from patients with psoriasis was compared with that of healthy individuals. Psoriatic skin scales, non-sterile and sterile, were tested for stimulatory effect on PMNs and ... More
Intracellular injections of permanent tracers in the fixed slice: a comparison of HRP and biocytin.
AuthorsColeman LA, Friedlander MJ
JournalJ Neurosci Methods
PubMed ID1474850
'Here we describe a method for intracellularly injecting mixtures of the fluorescent dye Lucifer Yellow and the permanent tracers HRP or biocytin into aldehyde-fixed slices of the dorsal lateral geniculate nucleus in young postnatal cats. Lucifer Yellow was used for visual control in the injection procedure and the inclusion of ... More
Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange.
AuthorsMcMaster GK, Carmichael GG
JournalProc Natl Acad Sci U S A
PubMed ID73185
'We have developed a simple and rapid system for the denaturation of nucleic acids and their subsequent analysis by gel electrophoresis. RNA and DNA are denatured in 1 M glyoxal (ethanedial) and 50% (vol/vol) dimethyl sulfoxide, at 50 degrees. The glyoxalated nucleic acids are then subjected to electrophoresis through either ... More
Characterization of yeast strains of the genera candida, Hansenula, Kluyveromyces, Pichia, Rhodotorula, and Saccharomyces by mixed-dye fluorometry.
AuthorsGeyer W, Brueggemann L, Flemming I, Nagel B
JournalInt J Syst Bacteriol
PubMed ID1854639
'An analytical method in which we used the selective adsorption of several fluorophores by yeast cells is described. The suitability of using binary mixtures of 1-pyrene butyric acid, 3,6-dimethylamino acridine, 4-acetamido-4''-isothiocyanatostilbene-2,2''-disulfonic acid, and rhodamine B isothiocyanate for the characterization and identification of microorganisms was tested with 98 yeast strains belonging ... More
Analysis of commercial samples of acridine orange using high-performance liquid chromatography.
AuthorsPaulsen RD, Nettleton GS
JournalJ Histochem Cytochem
PubMed ID3192936
'We analyzed commercial samples of acridine orange using reverse-phase high-performance liquid chromatography. The mobile phase of 90:10:2.5 acetonitrile:de-ionized water:pentane sulfonic acid gave baseline separations of components of acridine orange samples in 15 min. Many of the samples were fairly homogeneous; the absorbance due to the acridine orange component ranged from ... More