A cluster of noninvoluting endocytic cells at the margin of the zebrafish blastoderm marks the site of embryonic shield formation.
AuthorsCooper MS,D'Amico LA
JournalDevelopmental biology
PubMed ID8948584
Abnormal regulation of retinoic acid receptor beta2 expression and compromised allograft rejection in transgenic mice expressing antisense sequences to retinoic acid receptor beta1 and beta3.
Transgenic mice carrying antisense sequences common to the retinoic acid receptors (RAR) beta1 and beta3 were produced to examine roles of RARbeta1 and beta3 in the immune system. There were no significant changes of endogenous RARbeta1/beta3 expression at the mRNA level in T cells, B cells, and macrophages of the ... More
Analysis of the distribution of MRI contrast agents in the livers of small animals by means of complementary microscopies.
BACKGROUND: Magnetic resonance imaging (MRI) contrast agents contain magnetic molecules such as iron (Fe) or gadolinium (Gd) that are injected in vivo into rats or mice to study their distribution inside the liver. Fluorescent europium (Eu) can be used as a model of Gd to obtain comparable information of this ... More
Relationships among Bacterial Cell Size, Productivity, and Genetic Diversity in Aquatic Environments using Cell Sorting and Flow Cytometry.
AuthorsBernard L, Courties C, Servais P, Troussellier M, Petit M, Lebaron P
JournalMicrob Ecol
PubMed ID11029083
The study of relationships between cell size and productivity is of key importance in microbial ecology to understand which members of natural aquatic communities are responsible for the overall activity and/or productivity. Flow sorting of microorganisms from different environmental samples was used to analyze the activity of bacterial cells depending ... More
Systematic characterization of nuclear proteome during apoptosis: a quantitative proteomic study by differential extraction and stable isotope labeling.
AuthorsHwang SI, Lundgren DH, Mayya V, Rezaul K, Cowan AE, Eng JK, Han DK
JournalMol Cell Proteomics
PubMed ID16540461
'Identification and characterization of the nuclear proteome is important for detailed understanding of multiple signaling events in eukaryotic cells. Toward this goal, we extensively characterized the nuclear proteome of human T leukemia cells by sequential extraction of nuclear proteins with different physicochemical properties using three buffer conditions. This large scale ... More
Leishmania disease development depends on the presence of apoptotic promastigotes in the virulent inoculum.
Authorsvan Zandbergen G, Bollinger A, Wenzel A, Kamhawi S, Voll R, Klinger M, Müller A, Hölscher C, Herrmann M, Sacks D, Solbach W, Laskay T
JournalProc Natl Acad Sci U S A
PubMed ID16945916
'The obligate intracellular pathogen Leishmania major survives and multiplies in professional phagocytes. The evasion strategy to circumvent killing by host phagocytes and establish a productive infection is poorly understood. Here we report that the virulent inoculum of Leishmania promastigotes contains a high ratio of annexin A5-binding apoptotic parasites. This subpopulation ... More
Towards an understanding of apoptosis detection by SYTO dyes.
AuthorsWlodkowic D, Skommer J, Pelkonen J
JournalCytometry A
PubMed ID17200958
'BACKGROUND: SYTO probes are gaining momentum as reliable and easy to use markers of apoptotic cell death, but the phenomenon underlying reduced SYTO fluorescence in apoptotic cells as compared with normal cells is still not fully elucidated. Herein, we attempt to provide further insights into mechanisms of reduced SYTO16 fluorescence ... More
'Sorting of RNAs to specific subcellular loci occurs in diverse settings from fly oocytes to mammalian neurons. Using the membrane-permeable nucleic acid stain SYTO 14, we directly visualized the translocation of endogenous RNA in living cells. Labeled RNA was distributed nonrandomly as discrete granules in neuronal processes. The labeled granules ... More
Monitoring phytoplankton, bacterioplankton, and virioplankton in a coastal inlet (Bedford Basin) by flow cytometry.
AuthorsLi WK, Dickie PM
JournalCytometry
PubMed ID11429774
'BACKGROUND: To establish the prevailing state of the ecosystem for the assessment of long-term change, the abundance of microbial plankton in Bedford Basin (Nova Scotia, Canada) is monitored weekly by flow cytometry. METHODS: Phytoplankton are detected by their chlorophyll autofluorescence. Those that contain phycoerythrin are designated as Synechococcus cyanobacteria or ... More
Nucleic acid dyes for detection of apoptosis in live cells.
AuthorsFrey T
JournalCytometry
PubMed ID8582249
'Apoptotic thymocytes were found to be much dimmer than normal thymocytes when stained with several nucleic acid dyes. These dyes provide a quick and simple assay for apoptosis which works for live cells and does not require a UV laser. The collection of dyes giving this staining pattern includes reagents ... More
The FcgammaRII receptor triggers pp125FAK phosphorylation in platelets.
AuthorsHaimovich B, Regan C, DiFazio L, Ginalis E, Ji P, Purohit U, Rowley RB, Bolen J, Greco R
JournalJ Biol Chem
PubMed ID8663117
'Platelets express a single low affinity receptor for immunoglobulin, FcgammaRII, that triggers multiple cellular responses upon interaction with multivalent immune complexes. In this study we show that immobilized IgG is also a potent stimulant of platelet activation triggering adhesion, aggregation, massive dense granule secretion, and thromboxane production. Platelet adhesion to ... More
Phosphatidylserine-dependent engulfment by macrophages of nuclei from erythroid precursor cells.
AuthorsYoshida H, Kawane K, Koike M, Mori Y, Uchiyama Y, Nagata S
JournalNature
PubMed ID16193055
'Definitive erythropoiesis usually occurs in the bone marrow or fetal liver, where erythroblasts are associated with a central macrophage in anatomical units called ''blood islands''. Late in erythropoiesis, nuclei are expelled from the erythroid precursor cells and engulfed by the macrophages in the blood island. Here we show that the ... More
Effects of incubation temperature and time after thawing on viability assessment of peripheral hematopoietic progenitor cells cryopreserved for transplantation.
AuthorsYang H, Acker JP, Cabuhat M, McGann LE
JournalBone Marrow Transplant
PubMed ID14595390
'Three widely used viability assessments were compared: (1) membrane integrity of nucleated cells using trypan blue (TB) exclusion and a fluorometric membrane integrity assay (SYTO 13 and propidium iodide), (2) enumeration of viable CD34+ cells, and (3) clonogenic assay (granulocyte-macrophage colony-forming units, CFU-GM). Post thaw peripheral hematopoietic progenitor cells (HPC) ... More
Cleavage orientation and the asymmetric inheritance of Notch1 immunoreactivity in mammalian neurogenesis.
AuthorsChenn A, McConnell SK
JournalCell
PubMed ID7664342
'Neurons in the mammalian central nervous system are generated from progenitor cells near the lumen of the neural tube. Time-lapse microscopy of dividing cells in slices of developing cerebral cortex reveals that cleavage orientation predicts the fates of daughter cells. Vertical cleavages produce behaviorally and morphologically identical daughters that resemble ... More
Differential regulation of cardiomyocyte survival and hypertrophy by MDM2, an E3 ubiquitin ligase.
AuthorsToth A, Nickson P, Qin LL, Erhardt P
JournalJ Biol Chem
PubMed ID16339144
'MDM2 is an E3 ubiquitin ligase that regulates the proteasomal degradation and activity of proteins involved in cell growth and apoptosis, including the tumor suppressors p53 and retinoblastoma and the transcription factor E2F1. Although the effect of several MDM2 targets on cardiomyocyte survival and hypertrophy has already been investigated, the ... More
A rapid sample preparation technique for flow cytometric analysis of immunofluorescence allowing absolute enumeration of cell subpopulations.
AuthorsTerstappen LW, Meiners H, Loken MR
JournalJ Immunol Methods
PubMed ID2477460
'A simple and rapid method was developed for immunofluorescence measurements of cells by flow cytometry which does not require washing procedures, permitting absolute enumeration of cell subpopulations. Peripheral blood cells were labeled with fluorescein and phycoerythrin conjugated monoclonal antibodies and the nucleic acid stain LDS-751. Distilled water was added following ... More
Transmembrane inhibitors of P-glycoprotein, an ABC transporter.
AuthorsTarasova NI, Seth R, Tarasov SG, Kosakowska-Cholody T, Hrycyna CA, Gottesman MM, Michejda CJ
JournalJ Med Chem
PubMed ID15916428
'Drug resistance mediated by ABC transporters such as P-glycoprotein (P-gp) continues to be a major impediment to effective cancer chemotherapy. We have developed a panel of highly specific peptide inhibitors of P-gp based on the structure of the transmembrane domains of the transporter. These peptides are thought to exert their ... More
Oxidized LDLs induce massive apoptosis of cultured human endothelial cells through a calcium-dependent pathway. Prevention by aurintricarboxylic acid.
AuthorsEscargueil-Blanc I, Meilhac O, Pieraggi MT, Arnal JF, Salvayre R, Nègre-Salvayre A
JournalArterioscler Thromb Vasc Biol
PubMed ID9081689
'Oxidized LDLs are thought to play a central role in atherogenesis. Among their wide variety of biological properties, oxidized LDLs exhibit a cytotoxic effect on cultured vascular cells. Toxic doses of mildly oxidized LDLs elicited massive apoptosis in both primary and immortalized cultures of endothelial cells as shown by characteristic ... More
Manual punch versus power harvesting of osteochondral grafts.
AuthorsEvans PJ, Miniaci A, Hurtig MB
JournalArthroscopy
PubMed ID15007320
'PURPOSE: The purpose of the study was to analyze the effect on chondrocyte viability of 2 existing methods of harvesting osteochondral grafts used for articular cartilage resurfacing. TYPE OF STUDY: Acute animal experiment. METHODS: Power (P) trephine versus manual (M) punch harvesting was tested; 2.7-mm and 4.5-mm dowels were harvested ... More
Chromosome segregation defects contribute to aneuploidy in normal neural progenitor cells.
'Recent studies based predominantly on nucleotide hybridization techniques have identified aneuploid neurons and glia in the normal brain. To substantiate these findings and address how neural aneuploidy arises, we examined individual neural progenitor cells (NPCs) undergoing mitosis. Here we report the identification of chromosomal segregation defects in normal NPCs of ... More
Interaction of hepatitis C virus-like particles and cells: a model system for studying viral binding and entry.
AuthorsTriyatni M, Saunier B, Maruvada P, Davis AR, Ulianich L, Heller T, Patel A, Kohn LD, Liang TJ
JournalJ Virol
PubMed ID12186916
'Hepatitis C virus-like particles (HCV-LPs) containing the structural proteins of HCV H77 strain (1a genotype) was used as a model for HCV virion to study virus-cell interaction. HCV-LPs showed a buoyant density of 1.17 to 1.22 g/cm(3) in a sucrose gradient and formed double-shelled particles 35 to 49 nm in ... More
Neocentromere-mediated chromosome movement in maize.
AuthorsYu HG, Hiatt EN, Chan A, Sweeney M, Dawe RK
JournalJ Cell Biol
PubMed ID9362502
'Neocentromere activity is a classic example of nonkinetochore chromosome movement. In maize, neocentromeres are induced by a gene or genes on Abnormal chromosome 10 (Ab10) which causes heterochromatic knobs to move poleward at meiotic anaphase. Here we describe experiments that test how neocentromere activity affects the function of linked centromere/kinetochores ... More
Significance of size and nucleic acid content heterogeneity as measured by flow cytometry in natural planktonic bacteria.
AuthorsGasol JM, Zweifel UL, Peters F, Fuhrman JA, Hagström A
JournalAppl Environ Microbiol
PubMed ID10508078
'Total bacterial abundances estimated with different epifluorescence microscopy methods (4'',6-diamidino-2-phenylindole [DAPI], SYBR Green, and Live/Dead) and with flow cytometry (Syto13) showed good correspondence throughout two microcosm experiments with coastal Mediterranean water. In the Syto13-stained samples we could differentiate bacteria with apparent high DNA (HDNA) content and bacteria with apparent low ... More
The MspA porin promotes growth and increases antibiotic susceptibility of both Mycobacterium bovis BCG and Mycobacterium tuberculosis.
AuthorsMailaender C, Reiling N, Engelhardt H, Bossmann S, Ehlers S, Niederweis M
JournalMicrobiology
PubMed ID15073295
'Porins mediate the diffusion of hydrophilic solutes across the outer membrane of mycobacteria, but the efficiency of this pathway is very low compared to Gram-negative bacteria. To examine the importance of porins in slow-growing mycobacteria, the major porin MspA of Mycobacterium smegmatis was expressed in Mycobacterium tuberculosis and Mycobacterium bovis. ... More
AuthorsDosogne H, Vangroenweghe F, Mehrzad J, Massart-Leën AM, Burvenich C
JournalJ Dairy Sci
PubMed ID12703619
'Whereas many differential leukocyte count methods for high somatic cell count (SCC) milk from mastitic cows are available, only a few have been developed for low SCC milk. We have developed a flow cytometric differential leukocyte count method for low SCC milk. The procedure consists of 1) 1.5 ml of ... More
Coupling Bacterial Activity Measurements with Cell Sorting by Flow Cytometry.
AuthorsServais P, Courties C, Lebaron P, Troussellier M
JournalMicrob Ecol
PubMed ID10441710
'> Abstract A new procedure to investigate the relationship between bacterial cell size and activity at the cellular level has been developed; it is based on the coupling of radioactive labeling of bacterial cells and cell sorting by flow cytometry after SYTO 13 staining. Before sorting, bacterial cells were incubated ... More
Does the high nucleic acid content of individual bacterial cells allow us to discriminate between active cells and inactive cells in aquatic systems?
AuthorsLebaron P, Servais P, Agogué H, Courties C, Joux F
JournalAppl Environ Microbiol
PubMed ID11282632
'The nucleic acid contents of individual bacterial cells as determined with three different nucleic acid-specific fluorescent dyes (SYBR I, SYBR II, and SYTO 13) and flow cytometry were compared for different seawater samples. Similar fluorescence patterns were observed, and bacteria with high apparent nucleic acid contents (HNA) could be discriminated ... More
Modulation of [Ca2+]i signaling dynamics and metabolism by perinuclear mitochondria in mouse parotid acinar cells.
AuthorsBruce JI, Giovannucci DR, Blinder G, Shuttleworth TJ, Yule DI
JournalJ Biol Chem
PubMed ID14699167
'Parotid acinar cells exhibit rapid cytosolic calcium signals ([Ca2+]i) that initiate in the apical region but rapidly become global in nature. These characteristic [Ca2+]i signals are important for effective fluid secretion, which critically depends on a synchronized activation of spatially separated ion fluxes. Apically restricted [Ca2+]i signals were never observed ... More
SYTO16 labelling and flow cytometry of Mycobacterium avium.
AuthorsIbrahim P, Whiteley AS, Barer MR
JournalLett Appl Microbiol
PubMed ID9449859
'Mycobacterium avium cells were harvested from agar at different stages of their growth cycle, exposed to the minimum inhibitory concentration of isoniazid (INH) for 24 h and labelled with the fluorescent nucleic acid stain SYTO16. INH exposure led to a > 10-fold increase in the intensity of labelling in the ... More
A fluorescent Gram stain for flow cytometry and epifluorescence microscopy.
AuthorsMason DJ, Shanmuganathan S, Mortimer FC, Gant VA
JournalAppl Environ Microbiol
PubMed ID9647848
'The fluorescent nucleic acid binding dyes hexidium iodide (HI) and SYTO 13 were used in combination as a Gram stain for unfixed organisms in suspension. HI penetrated gram-positive but not gram-negative organisms, whereas SYTO 13 penetrated both. When the dyes were used together, gram-negative organisms were rendered green fluorescent by ... More
Calmodulin, gametes and fertilisation.
AuthorsCourtot AM, Pesty A, Lefèvre B
JournalZygote
PubMed ID10418102
'The role of calmodulin in fertilisation events was examined in a zona-free mouse system by using a selective calmodulin inhibitor, calmidazolium (1 microM). The effects of this antagonist were studied either on the ooplasmic calcium oscillations induced by fertilisation by using the Ca2+ indicator, fluo-3/AM, or on pronucleus formation 4 ... More
Suppression of Fas/APO-1-mediated apoptosis by mitogen-activated kinase signaling.
AuthorsHolmström TH, Chow SC, Elo I, Coffey ET, Orrenius S, Sistonen L, Eriksson JE
JournalJ Immunol
PubMed ID9510160
'Jurkat T cells undergo rapid apoptosis upon stimulation of the Fas/APO-1 (CD95) receptor. We examined the role of the mitogen-activated protein kinase (MAPK) cascade as a negative regulator of Fas-mediated apoptosis. To this end, we used both physiologic and artificial activators of MAPK, all of which activate MAPK by distinct ... More
Assessment of the effects of gramicidin, formaldehyde, and surfactants on Escherichia coli by flow cytometry using nucleic acid and membrane potential dyes.
AuthorsComas J, Vives-Rego J
JournalCytometry
PubMed ID9298812
'Two membrane potential sensitive dyes (Rhodamine 123 and bis-oxonol) and three nucleic acid dyes (propidium iodide, SYTO-13, and SYTO-17) were used to assess the effect of surfactants on Escherichia coli. The ability of E. coli to be stained by these probes was validated at different physiological states. Propidium iodide was ... More
Identification of an in vivo inhibitor of Bacillus anthracis spore germination.
AuthorsAkoachere M, Squires RC, Nour AM, Angelov L, Brojatsch J, Abel-Santos E
JournalJ Biol Chem
PubMed ID17296608
'Germination of Bacillus anthracis spores into the vegetative form is an essential step in anthrax pathogenicity. This process can be triggered in vitro by the common germinants inosine and alanine. Kinetic analysis of B. anthracis spore germination revealed synergy and a sequential mechanism between inosine and alanine binding to their ... More
Integrating conflicting chemotactic signals. The role of memory in leukocyte navigation.
AuthorsFoxman EF, Kunkel EJ, Butcher EC
JournalJ Cell Biol
PubMed ID10545501
'Leukocytes navigate through complex chemoattractant arrays, and in so doing, they must migrate from one chemoattractant source to another. By evaluating directional persistence and chemotaxis during neutrophil migration under agarose, we show that cells migrating away from a local chemoattractant, against a gradient, display true chemotaxis to distant agonists, often ... More
Differential contribution of cytoplasmic Ca2+ and Ca2+ influx to gamete fusion and egg activation in maize.
AuthorsAntoine AF, Faure JE, Dumas C, Feijó JA
JournalNat Cell Biol
PubMed ID11781574
'In multicellular organisms, gamete fusion triggers a set of events, collectively known as egg activation, that leads to the development of a new individual. Every species that has been studied shows at least one rise in cytoplasmic Ca2+ concentration ([Ca2+]Cyt) after gamete fusion which is believed to be involved in ... More
Proteins with glycosylphosphatidylinositol (GPI) signal sequences have divergent fates during a GPI deficiency. GPIs are essential for nuclear division in Trypanosoma cruzi.
AuthorsGarg N, Tarleton RL, Mensa-Wilmot K
JournalJ Biol Chem
PubMed ID9139697
'Glycosylphosphatidylinositols (GPIs) are membrane anchors for cell surface proteins of several major protozoan parasites of humans, including Trypanosoma cruzi, the causative agent of Chagas'' disease. To investigate the general role of GPIs in T. cruzi, we generated GPI-deficient parasites by heterologous expression of T. brucei GPI-phospholipase C. Putative protein-GPI intermediates ... More
Detection of apoptosis in live cells by MitoTracker red CMXRos and SYTO dye flow cytometry.
AuthorsPoot M, Gibson LL, Singer VL
JournalCytometry
PubMed ID9098628
'We characterized the ability of six SYTO nucleic acid stains and a mitochondrial stain to resolve by flow cytometry camptothecin-induced apoptotic and non-apoptotic cells. Staining live human lymphoid B-cells showed such resolution with SYTO 11, 12, 13, 14, and 16 dyes. H9, HL-60, and Jurkat cells did not show resolution ... More
Resolution of mitotic cells using laser scanning cytometry.
AuthorsLuther E, Kamentsky LA
JournalCytometry
PubMed ID8900469
'A microscope-based laser scanning cytometer (LSCM) has been developed that automatically measures multiple wavelength fluorescence and light scattering of cells on a microscope slide and generates lists of cytochemical and morphological features for each of thousands of cells in a typical sample. For a sample stained with a DNA stain, ... More
Characteristics of different nucleic acid staining dyes for DNA fragment sizing by flow cytometry.
AuthorsYan X, Grace WK, Yoshida TM, Habbersett RC, Velappan N, Jett JH, Keller RA, Marrone BL
JournalAnal Chem
PubMed ID10624155
'An efficient and reliable double-stranded DNA (dsDNA) staining protocol for DNA fragment sizing by flow cytometry is presented. The protocol employs 0.8 microM of PicoGreen to label a wide range of DNA concentrations (0.5 ng/mL to 10,000 ng/mL) without regard to the solution dye/bp ratios and without initial quantification of ... More
Microtubule-dependent recruitment of Staufen-green fluorescent protein into large RNA-containing granules and subsequent dendritic transport in living hippocampal neurons.
AuthorsKöhrmann M, Luo M, Kaether C, DesGroseillers L, Dotti CG, Kiebler MA
JournalMol Biol Cell
PubMed ID10473638
'Dendritic mRNA transport and local translation at individual potentiated synapses may represent an elegant way to form synaptic memory. Recently, we characterized Staufen, a double-stranded RNA-binding protein, in rat hippocampal neurons and showed its presence in large RNA-containing granules, which colocalize with microtubules in dendrites. In this paper, we transiently ... More
Comparison of blue nucleic acid dyes for flow cytometric enumeration of bacteria in aquatic systems.
AuthorsLebaron P, Parthuisot N, Catala P
JournalAppl Environ Microbiol
PubMed ID9572943
'Seven blue nucleic acid dyes from Molecular Probes Inc. (SYTO-9, SYTO-11, SYTO-13, SYTO-16, SYTO-BC, SYBR-I and SYBR-II) were compared with the DAPI (4'',6-diamidino-2-phenylindole) method for flow cytometric enumeration of live and fixed bacteria in aquatic systems. It was shown that SYBR-II and SYTO-9 are the most appropriate dyes for bacterial ... More
Flow cytometry counting of CD34+ cells in whole blood.
AuthorsFornas O, Garcia J, Petriz J
JournalNat Med
PubMed ID10888936
Analyzing morphogenetic cell behaviors in vitally stained zebrafish embryos.
AuthorsCooper MS, D'Amico LA, Henry CA
JournalMethods Mol Biol
PubMed ID10231793
Use of confocal microscopy to investigate cell structure and function.
AuthorsBkaily G, Jacques D, Pothier P
JournalMethods Enzymol
PubMed ID10506971
Use of nucleic acid dyes SYTO-13, TOTO-1, and YOYO-1 in the study of Escherichia coli and marine prokaryotic populations by flow cytometry.
AuthorsGuindulain T, Comas J, Vives-Rego J
JournalAppl Environ Microbiol
PubMed ID9361447
Three nucleic acid dyes (SYTO-13, TOTO-1, and YOYO-1) were tested on cultures of Escherichia coli and marine prokaryote populations. These dyes stain the RNA and DNA in E. coli but only respond to DNA in marine populations, according to the histograms obtained after DNase and RNase treatments. ... More
SYTO dyes and histoproteins--myriad of applications.
AuthorsTárnok A,
JournalCytometry A
PubMed ID18496838
SYTO dyes differ from each other in one or more characteristics, including cell permeability, ?uorescence enhancement upon binding nucleic acids, excitation and emission spectra, DNA/RNA selectivity and binding af?nity. This makes them versatile tools for cell biologists. ... More
The use of confocal microscopy in the investigation of cell structure and function in the heart, vascular endothelium and smooth muscle cells.
AuthorsBkaily G, Pothier P, D'Orléans-Juste P, Simaan M, Jacques D, Jaalouk D, Belzile F, Hassan G, Boutin C, Haddad G, Neugebauer W
JournalMol Cell Biochem
PubMed ID9278244
In recent years, fluorescence microscopy imaging has become an important tool for studying cell structure and function. This non invasive technique permits characterization, localisation and qualitative quantification of free ions, messengers, pH, voltage and a pleiad of other molecules constituting living cells. In this paper, we present results using various ... More
Optical sensors for detection of bacteria. 1. General concepts and initial development.
AuthorsChuang H, Macuch P, Tabacco MB
JournalAnal Chem
PubMed ID11217747
The concept of using immobilized nucleic acid stains as detection chemistry to fabricate optical bacterial sensors is first demonstrated. SYTO 13 (a green fluorescent cell stain) is used as the molecular recognition element and fluorescent reporter in the sensor. The sensor responds to aqueous and aerosolized bacterial samples in 15 ... More
Flow cytometric analysis of a marine LAS-degrading consortia.
The specific nucleic acid fluorochrome SYTO-13 was used in flow cytometric analysis to assess changes in the density and heterogeneity of marine bacterial populations which biodegrade linear alkylbenzene sulphonate (LAS). Seawater samples with LAS and incubated in the laboratory (20 degrees C, 100 rpm, 30 days) were used to monitor ... More
Full developmental potential of mammalian preimplantation embryos is maintained after imaging using a spinning-disk confocal microscope.
AuthorsRoss PJ, Perez GI, Ko T, Yoo MS, Cibelli JB
JournalBiotechniques
PubMed ID17191620
Fluorescent live imaging of cells and embryos at subcellular resolution poses significant challenges for biologists due to morbidity and mortality ensuing from phototoxicity. Here we report the use of a spinning-disk confocal microscope to image mouse and bovine preimplantation embryos without impairing their developmental potential. We also present data indicating ... More
Assessment of fluorochromes for two-photon laser scanning microscopy of biofilms.
AuthorsNeu TR, Kuhlicke U, Lawrence JR
JournalAppl Environ Microbiol
PubMed ID11823234
A major limitation for the use of two-proton laser scanning microscopy (2P-LSM) in biofilm and other studies is the lack of a thorough understanding of the excitation-emission responses of potential fluorochromes. In order to use 2P-LSM, the utility of various fluorochromes and probes specific for a range of biofilm constituents ... More
NUC-1, a caenorhabditis elegans DNase II homolog, functions in an intermediate step of DNA degradation during apoptosis.
AuthorsWu YC, Stanfield GM, Horvitz HR
JournalGenes Dev
PubMed ID10716942
One hallmark of apoptosis is the degradation of chromosomal DNA. We cloned the Caenorhabditis elegans gene nuc-1, which is involved in the degradation of the DNA of apoptotic cells, and found that nuc-1 encodes a homolog of mammalian DNase II. We used the TUNEL technique to assay DNA degradation in ... More
Doublecortin is a developmentally regulated, microtubule-associated protein expressed in migrating and differentiating neurons.
AuthorsFrancis F, Koulakoff A, Boucher D, Chafey P, Schaar B, Vinet MC, Friocourt G, McDonnell N, Reiner O, Kahn A, McConnell SK, Berwald-Netter Y, Denoulet P, Chelly J
JournalNeuron
PubMed ID10399932
Recently, we and others reported that the doublecortin gene is responsible for X-linked lissencephaly and subcortical laminar heterotopia. Here, we show that Doublecortin is expressed in the brain throughout the period of corticogenesis in migrating and differentiating neurons. Immunohistochemical studies show its localization in the soma and leading processes of ... More
Neurotrophin-3 signals redistribute RNA in neurons.
AuthorsKnowles RB, Kosik KS
JournalProc Natl Acad Sci U S A
PubMed ID9405694
The translocation of specific mRNAs to dendrites and their potential for locally regulated translation are likely to serve as an effector in neuronal plasticity. Whether translation in dendrites is regulated by delivery of the RNA to sites of plasticity or a stationary pool of localized RNA undergoes enhanced translational efficiency ... More
Novel model for multispecies biofilms that uses rigid gas-permeable lenses.
Oral biofilms comprise complex multispecies consortia aided by specific inter- and intraspecies interactions occurring among commensals and pathogenic bacterial species. Oral biofilms are primary initiating factors of periodontal disease, although complex multifactorial biological influences, including host cell responses, contribute to the individual outcome of the disease. To provide a system ... More
SYTO probes in the cytometry of tumor cell death.
AuthorsWlodkowic D, Skommer J, Darzynkiewicz Z,
JournalCytometry A
PubMed ID18260152
Apoptosis is a complex and finely controlled cell death process of great relevance in tissue homeostasis and pathogenesis. The majority of classical apoptotic features can be examined by flow as well as image cytometry. Therefore, cytometry has been used as a technology of choice in studies of tumor cell demise. ... More
Glutamate-induced neuronal death: a succession of necrosis or apoptosis depending on mitochondrial function.
AuthorsAnkarcrona M, Dypbukt JM, Bonfoco E, Zhivotovsky B, Orrenius S, Lipton SA, Nicotera P
JournalNeuron
PubMed ID7576644
During ischemic brain injury, glutamate accumulation leads to overstimulation of postsynaptic glutamate receptors with intracellular Ca2+ overload and neuronal cell death. Here we show that glutamate can induce either early necrosis or delayed apoptosis in cultures of cerebellar granule cells. During and shortly after exposure to glutamate, a subpopulation of ... More
Quantification of neurotoxicity and identification of cellular subsets in a three-dimensional brain model.
AuthorsPulliam L, Stubblebine M, Hyun W
JournalCytometry
PubMed ID9581626
Imaging of cells in a large intact three-dimensional tissue remains difficult. Quantification and identification of cell damage in a mixed culture system has been limited by the inability of fluorescent probes to discriminate types of cellular death and penetrate tissue more that 100 microm thick. We have investigated several probes ... More
EGR2 mutations in inherited neuropathies dominant-negatively inhibit myelin gene expression.
AuthorsNagarajan R, Svaren J, Le N, Araki T, Watson M, Milbrandt J
JournalNeuron
PubMed ID11394999
The identification of EGR2 mutations in patients with neuropathies and the phenotype Egr2/Krox20(-/-) have demonstrated that the Egr2 transcription factor is critical for peripheral nerve myelination. However, the mechanism by which these mutations cause disease remains unclear, as most patients present with disease in the heterozygous state, whereas Egr2(+/-) mice ... More
In vivo identification of ribonucleoprotein-RNA interactions.
AuthorsZielinski J, Kilk K, Peritz T, Kannanayakal T, Miyashiro KY, Eiríksdóttir E, Jochems J, Langel U, Eberwine J
JournalProc Natl Acad Sci U S A
PubMed ID16432185
To understand the role of RNA-binding proteins (RBPs) in the regulation of gene expression, methods are needed for the in vivo identification of RNA-protein interactions. We have developed the peptide nucleic acid (PNA)-assisted identification of RBP technology to enable the identification of proteins that complex with a target RNA in ... More
A role for a rat homolog of staufen in the transport of RNA to neuronal dendrites.
AuthorsTang SJ, Meulemans D, Vazquez L, Colaco N, Schuman E
JournalNeuron
PubMed ID11709157
RNAs are present in dendrites and may be used for local protein synthesis in response to synaptic activity. To begin to understand dendritic RNA targeting, we cloned a rat homolog of staufen, a Drosophila gene that participates in mRNA targeting during development. In hippocampal neurons, rat staufen protein displays a ... More
An immunohistochemical method for the detection of proteins in the vertebrate lens.
AuthorsReed NA, Oh DJ, Czymmek KJ, Duncan MK
JournalJ Immunol Methods
PubMed ID11384685
Fluorescence immunohistochemistry has traditionally been difficult or impossible to perform on the vertebrate lens because of its extremely high protein content. Described here is a robust and rapid method for preparing and labeling vertebrate eyes for confocal microscopy. This technique has successfully been applied to localize proteins in the lens ... More
Delineation of type I protein kinase A-selective signaling events using an RI anchoring disruptor.
AuthorsCarlson CR, Lygren B, Berge T, Hoshi N, Wong W, Taskén K, Scott JD
JournalJ Biol Chem
PubMed ID16728392
Control of specificity in cAMP signaling is achieved by A-kinase anchoring proteins (AKAPs), which assemble cAMP effectors such as protein kinase A (PKA) into multiprotein signaling complexes in the cell. AKAPs tether the PKA holoenzymes at subcellular locations to favor the phosphorylation of selected substrates. PKA anchoring is mediated by ... More
Electrophoretic behavior of individual nuclear species as determined by capillary electrophoresis with laser-induced fluorescence detection.
AuthorsGunasekera N, Musier-Forsyth K, Arriaga E
JournalElectrophoresis
PubMed ID12210266
We determined the feasibility of using capillary electrophoresis with postcolumn laser-induced fluorescence (CE-LIF) detection to characterize electrophoretic properties of isolated cell nuclei and impurities present in nuclear fractions. These fractions were isolated from NS-1 mouse hybridoma cells, stained with hexidium iodide, a DNA intercalating dye, and analyzed by CE-LIF detection. ... More
Cytoskeletal breakdown and apoptosis elicited by NO donors in cerebellar granule cells require NMDA receptor activation.
AuthorsBonfoco E, Leist M, Zhivotovsky B, Orrenius S, Lipton SA, Nicotera P
JournalJ Neurochem
PubMed ID8931482
We have recently demonstrated that nitric oxide (NO) donors can trigger either apoptosis or necrosis of neurons as a function of the intensity of the exposure. Here, we show that the apoptosis induced by the NO donors S-nitrosocysteine (SNOC) or S-nitroso-N-acetyl-penicillamine (SNAP) in cultured cerebellar granule cells (CGCs) depends on ... More
Flow cytometry assessment of bacterioplankton in tropical marine environments.
AuthorsAndrade L, Gonzalez AM, Araujo FV, Paranhos R
JournalJ Microbiol Methods
PubMed ID14607430
Flow cytometry was used to characterize bacterioplankton from two tropical environments in Brazil: the eutrophic Guanabara Bay and the oligotrophic southwest Atlantic Ocean. Bacterial abundance was evaluated by flow cytometry, and cells were stained with SYTO 13, allowing demonstration of differences in nucleic acid content. Bacterial production was also evaluated ... More
Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development.
AuthorsChen H, Detmer SA, Ewald AJ, Griffin EE, Fraser SE, Chan DC
JournalJ Cell Biol
PubMed ID12527753
Mitochondrial morphology is determined by a dynamic equilibrium between organelle fusion and fission, but the significance of these processes in vertebrates is unknown. The mitofusins, Mfn1 and Mfn2, have been shown to affect mitochondrial morphology when overexpressed. We find that mice deficient in either Mfn1 or Mfn2 die in midgestation. ... More
Confocal imaging of microglial cell dynamics in hippocampal slice cultures.
AuthorsDailey ME, Waite M
JournalMethods
PubMed ID10356354
Methods are described for imaging the cellular dynamics of microglia in live mammalian brain slice cultures. Brain slices prepared from developing rat hippocampus are cultured for up to 2 weeks by the roller tube or static filter culture technique, stained with one or more fluorescent dyes, and imaged by scanning ... More
Discrimination of DNA and RNA in cells by a vital fluorescent probe: lifetime imaging of SYTO13 in healthy and apoptotic cells.
Authorsvan Zandvoort MA, de Grauw CJ, Gerritsen HC, Broers JL, oude Egbrink MG, Ramaekers FC, Slaaf DW
JournalCytometry
PubMed ID11933012
BACKGROUND: Of the few vital DNA and RNA probes, the SYTO dyes are the most specific for nucleic acids. However, they show no spectral contrast upon DNA or RNA binding. We show that fluorescence lifetime imaging using two-photon excitation of SYTO13 allows differential and simultaneous imaging of DNA and RNA ... More
Detection of changes occurring during recovery from the dauer stage in Heterorhabditis bacteriophora.
AuthorsDolan KM, Jones JT, Burnell AM
JournalParasitology
PubMed ID12166523
Nematodes of the genus Heterorhabditis are insect parasites that are widely used as biological control agents. When conditions are unfavourable for reproduction in H. bacteriophora, a long-lived, non-feeding, survival and dispersal stage, the dauer juvenile (DJ), is formed. This DJ stage is also adapted for host finding and infection. When ... More
A multicolor, no-lyse no-wash assay for the absolute counting of CD34+ cells by flow cytometry.
AuthorsAlvarez-Larran A, Jover L, Marin P, Petriz J
JournalCytometry
PubMed ID12360574
BACKGROUND: We previously developed a method for counting CD34(+) cells in unlysed whole blood. This method was applied to normal human bone marrow, peripheral blood after mobilization of progenitor cells, leukapheresis products, and cord blood and was validated with two different lyse-no wash methods. However, the main advantage that we ... More
Kinetics and mechanism of intercellular ice propagation in a micropatterned tissue construct.
AuthorsIrimia D, Karlsson JO
JournalBiophys J
PubMed ID11916845
Understanding the effects of cell-cell interaction on intracellular ice formation (IIF) is required to design optimized protocols for cryopreservation of tissue. To determine the effects of cell-cell interactions during tissue freezing, without confounding effects from uncontrolled factors (such as time in culture, cell geometry, and cell-substrate interactions), HepG2 cells were ... More
Involvement of RNA and DNA in the staining of Escherichia coli by SYTO 13.
AuthorsGuindulain T, Vives-Rego J
JournalLett Appl Microbiol
PubMed ID11874539
AIMS: To assess the extent to which DNA and RNA bacterial content contributes to fluorescent response of SYTO 13. METHODS AND RESULTS: RNA and DNA of Escherichia coli 536 cells were extracted and fluorimetrically quantified to compare the different contents, throughout a 24 h culture, with their SYTO 13 fluorescence ... More
Flow cytometry to evaluate Theileria sergenti parasitemia using the fluorescent nucleic acid stain, SYTO16.
AuthorsYagi Y, Shiono H, Kurabayashi N, Yoshihara K, Chikayama Y
JournalCytometry
PubMed ID11042620
BACKGROUND: Although the infection of Theileria sergenti is demonstrated by intraerythrocytic localization of this parasite, much time and labor are necessary in order to determine this. We applied flow cytometry to evaluate T. sergenti parasitemia using the fluorescent nucleic acid stain method. METHODS: Peripheral blood samples from cattle infected with ... More
Quantification of T-cell-mediated apoptosis in heterogeneous leukemia populations using four-color multiparameter flow cytometry.
AuthorsWesters TM, Houtenbos I, Schuurhuis GJ, Ossenkoppele GJ, van de Loosdrecht AA
JournalCytometry A
PubMed ID15895438
BACKGROUND: The unique capacity of dendritic cells to present antigens to naive T cells is being increasingly utilized in cancer therapy. The efficacy of cell-based immunotherapy can be analyzed by determination of cytotoxic activity of T cells toward tumor cells in vitro. This study supplies a flow cytometric method to ... More
Rac1 and Cdc42 regulate hyphal growth and cytokinesis in the dimorphic fungus Ustilago maydis.
AuthorsMahlert M, Leveleki L, Hlubek A, Sandrock B, Bölker M
JournalMol Microbiol
PubMed ID16390450
Small GTP-binding proteins of the highly conserved Rho family act as molecular switches regulating cell signalling, cytoskeletal organization and vesicle trafficking in eukaryotic cells. Here we show that in the dimorphic plant pathogenic fungus Ustilago maydis deletion of either cdc42 or rac1 results in loss of virulence but does not ... More
Arrays of self-assembled monolayers for studying inhibition of bacterial adhesion.
This paper describes a simple and convenient method for the rapid screening of potential inhibitors of bacterial adhesion and for the quantitative evaluation of the efficacy of the inhibitors using arrays of self-assembled monolayers (SAMs) of alkanethiolates on gold that are presented on a 96-well microtiter plate. The SAMs present ... More
Nuclear and cytosolic calcium changes in osteoclasts stimulated with ATP and integrin-binding peptide.
AuthorsParkinson N, Bolsover S, Mason W
JournalCell Calcium
PubMed ID9883275
Cytosolic calcium modulates the activity of osteoclasts, large multinucleate cells that resorb bone. Nuclear events, such as gene transcription, are also calcium-regulated in these cells, and fluorescence imaging has suggested that calcium signals produced by some stimuli are specifically targeted to, or amplified within, osteoclast nuclei. We used two alternative ... More
Functional cardiac cell constructs on cellulose-based scaffolding.
AuthorsEntcheva E, Bien H, Yin L, Chung CY, Farrell M, Kostov Y
JournalBiomaterials
PubMed ID15147821
Cellulose and its derivatives have been successfully employed as biomaterials in various applications, including dialysis membranes, diffusion-limiting membranes in biosensors, in vitro hollow fibers perfusion systems, surfaces for cell expansion, etc. In this study, we tested the potential of cellulose acetate (CA) and regenerated cellulose (RC) scaffolds for growing functional ... More
Intercellular ice propagation: experimental evidence for ice growth through membrane pores.
AuthorsAcker JP, Elliott JA, McGann LE
JournalBiophys J
PubMed ID11509353
Propagation of intracellular ice between cells significantly increases the prevalence of intracellular ice in confluent monolayers and tissues. It has been proposed that gap junctions facilitate ice propagation between cells. This study develops an equation for capillary freezing-point depression to determine the effect of temperature on the equilibrium radius of ... More
Two-photon microscopy of vital murine elastic and muscular arteries. Combined structural and functional imaging with subcellular resolution.
AuthorsMegens RT, Reitsma S, Schiffers PH, Hilgers RH, De Mey JG, Slaaf DW, oude Egbrink MG, van Zandvoort MA
JournalJ Vasc Res
PubMed ID17192719
Understanding vascular pathologies requires insight in the structure and function, and, hence, an imaging technique combining subcellular resolution, large penetration depth, and optical sectioning. We evaluated the applicability of two-photon laser-scanning microscopy (TPLSM) in large elastic and small muscular arteries under physiological conditions. Elastic (carotid) and muscular (uterine, mesenteric) arteries ... More
Cellular effects of monohydrochloride of L-arginine, N-lauroyl ethylester (LAE) on exposure to Salmonella typhimurium and Staphylococcus aureus.
AuthorsRodríguez E, Seguer J, Rocabayera X, Manresa A
JournalJ Appl Microbiol
PubMed ID15078505
AIMS: Here we study the effect of monohydrochloride of L-arginine, N(alpha)-lauroyl ethylester (LAE), a cationic preservative derived from lauric acid and arginine, on the cell envelopes of Salmonella typhimurium and Staphylococcus aureus at sub-lethal concentration such as their respective minimal inhibitory concentrations, 32 and 8 microg ml(-1), respectively. METHODS AND ... More
Safe susceptibility testing of Mycobacterium tuberculosis by flow cytometry with the fluorescent nucleic acid stain SYTO 16.
AuthorsPina-Vaz C, Costa-de-Oliveira S, Rodrigues AG
JournalJ Med Microbiol
PubMed ID15591259
The time needed to obtain susceptibility results of Mycobacterium tuberculosis using classical methodologies is still too long, and flow cytometry is a promising technique in the setting of the clinical laboratory, giving fast results. A safe, reliable and rapid method to study susceptibility to streptomycin, isoniazide, rifampicin and ethambutol is ... More
Formation of planar and spiral Ca2+ waves in isolated cardiac myocytes.
AuthorsIshida H, Genka C, Hirota Y, Nakazawa H, Barry WH
JournalBiophys J
PubMed ID10512831
A novel Nipkow-type confocal microscope was applied to image spontaneously propagating Ca2+ waves in isolated rat ventricular myocytes by means of fluo-3. The sarcolemma was imaged with di-8-ANEPPS and the nucleus with SYTO 11. Full frame images in different vertical sections were obtained at video frame rate by means of ... More
Determination of in situ bacterial growth rates in aquifers and aquifer sediments.
AuthorsMailloux BJ, Fuller ME
JournalAppl Environ Microbiol
PubMed ID12839747
Laboratory and field-scale studies with stained cells were performed to monitor cell growth in groundwater systems. During cell division, the fluorescence intensity of the protein stain 5-(and 6-)-carboxyfluorescein diacetate succinimidyl ester (CFDA/SE) for each cell is halved, and the intensity can be tracked with a flow cytometer. Two strains of ... More
A double-label technique that monitors sulfur mustard damage to nuclei and mitochondria of normal human epidermal keratinocytes in vitro.
AuthorsCook JR, Van Buskirk RG
JournalToxicol Pathol
PubMed ID9323838
Sulfur mustard and 2-chloro ethyl ethyl sulfide (CEES, a sulfur mustard analog) is known to have immediate (minutes), long-term (hours to days), and toxic effects on human skin. Research was directed toward developing a single in vitro assay that might reflect both these short-term and long-term effects of this vesicating ... More
Characterization of bimodal cell death of insect cells in a rotating-wall vessel and shaker flask
In previous publications, we reported the benefits of a high-aspect rotating-wall vessel (HARV) over conventional bioreactors for insect-cell cultivation in terms of reduced medium requirements and enhanced longevity. To more fully understand the effects that HARV cultivation has on longevity, the present study characterizes the mode and kinetics of Spodoptera ... More
Fluorescence lifetime-based discrimination and quantification of cellular DNA and RNA with phase-sensitive flow cytometry.
AuthorsCui HH, Valdez JG, Steinkamp JA, Crissman HA
JournalCytometry A
PubMed ID12596251
BACKGROUND: Simultaneous measurement of cellular DNA and RNA content provides information for determination of the functional status of cells and, clinically, for the diagnosis and grading assessment of various tumors. Most current flow cytometric methods are based on resolving the fluorescence emission spectra of dyes that bind preferentially to either ... More
Highly sensitive and specific detection of P-glycoprotein function for haematological and solid tumour cells using a novel nucleic acid stain.
AuthorsBroxterman HJ, Schuurhuis GJ, Lankelma J, Oberink JW, Eekman CA, Claessen AM, Hoekman K, Poot M, Pinedo HM
JournalBr J Cancer
PubMed ID9376262
Progress in our understanding of the contribution of P-glycoprotein (P-gp)-mediated resistance to chemotherapy failure in haematological as well as solid tumours has been hampered by the lack of highly sensitive, reliable methods for the detection of P-gp function in fresh human tumour cells. The present study identifies the novel nucleic ... More
Colocalization of single ribosomes with intermediate filaments in puromycin-treated and serum-starved mouse embryo fibroblasts.
Previous experiments have revealed a relatively weak electrostatic binding capacity of in vitro reconstituted intermediate filaments (IFs) as well as of natural IFs of whole cell mount preparations for purified ribosomal particles of mammalian origin. In order to demonstrate that such associations also occur in vivo, intact cells were subjected ... More
Bradykinin activates R-, T-, and L-type Ca2+ channels and induces a sustained increase of nuclear Ca2+ in aortic vascular smooth muscle cells.
AuthorsBkaily G, Jaalouk D, Jacques D, Economos D, Hassan G, Simaan M, Regoli D, Pothier P
JournalCan J Physiol Pharmacol
PubMed ID9276144
The mechanism(s) fo Ca2+ entry stimulated by bradykinin (BK) and the receptor subtype responsible for this effect were examined in human and rabbit aortic vascular smooth muscle cells (VSMCs). Using the whole-cell voltage clamp technique, BK (10(-6)M) significantly (p < 0.05) increased both T- and L-type Ca2+ currents (ICa) in ... More
Large populations of non-clonogenic early apoptotic CD34-positive cells are present in frozen-thawed peripheral blood stem cell transplants.
AuthorsSchuurhuis GJ, Muijen MM, Oberink JW, de Boer F, Ossenkoppele GJ, Broxterman HJ
JournalBone Marrow Transplant
PubMed ID11313682
Apoptosis is the common cell death pathway which is initiated by a variety of different stimuli. The recognition of early apoptotic events would markedly improve reliability and convenience of apoptosis assays. In the present study the vital stain SytoR 16 in combination with the permeability marker 7-amino actinomycin D, (7-AAD) ... More
Confocal microscopic analysis of morphogenetic movements.
AuthorsCooper MS, D'Amico LA, Henry CA
JournalMethods Cell Biol
PubMed ID9891361
Confocal microscopy is an excellent means of imaging cellular dynamics within living zebrafish embryos because it provides a means of optically sectioning tissues that have been labeled with specific fluorescent probe molecules. In order to study genetically encoded patterns of cell behavior that are involved in the formation of germ ... More
Options of flow cytometric three-colour DNA measurements to quantitate EGFR in subpopulations of human bladder cancer.
AuthorsBrockhoff G, Endl E, Minuth W, Hofstädter F, Knüchel R
JournalAnal Cell Pathol
PubMed ID8844105
Flow cytometric multi-parameter analysis has proven to be a powerful tool to characterize subpopulations of cell suspensions, and is applied routinely in hematology. However, in studies of cancer where there is interest in defining phenotypic markers in conjunction with DNA content, this method has hardly been applied [6]. Our objective ... More
Overexpression of PAX6(5a) in lens fiber cells results in cataract and upregulation of (alpha)5(beta)1 integrin expression.
AuthorsDuncan MK, Kozmik Z, Cveklova K, Piatigorsky J, Cvekl A
JournalJ Cell Sci
PubMed ID10954416
The PAX6 gene, a key regulator of eye development, produces two major proteins that differ in paired domain structure: PAX6 and PAX6(5a). It is known that an increase in the PAX6(5a) to PAX6 ratio leads to multiple ocular defects in humans. Here, transgenic mice were created that overexpress human PAX6(5a) ... More
Discrimination of live and early apoptotic mononuclear cells by the fluorescent SYTO 16 vital dye.
AuthorsSparrow RL, Tippett E
JournalJ Immunol Methods
PubMed ID16165150
Accurate detection of apoptotic cells is important for the determination of cell viability. The aim of this study was to compare the sensitivity of the cell permeant SYTO 16 fluorescent dye for detecting early apoptotic mononuclear cells (MNCs) in normal donor blood with other apoptosis assays [i.e. Annexin-V, light scatter/7-amino-actinomycin-D ... More
Nuclear Ca2+ signaling to endothelin-1 in rat aortic smooth-muscle cells.
AuthorsNaik RD, McNeill JR, Wilson TW, Gopalakrishnan V
JournalJ Cardiovasc Pharmacol
PubMed ID9595437
Vascular smooth-muscle cells (VSMCs) isolated from genetically hypertensive animals show increased intracellular free calcium levels ([Ca2+]i) in response to endothelin-1 (ET-1). The differences in time course and distribution of Ca2+ increase after addition of ET-1 within the VSMCs are unknown. Therefore, ET-1-evoked changes in fluo-3 fluorescence were determined using a ... More
Three restricted forms of Epstein-Barr virus latency counteracting apoptosis in c-myc-expressing Burkitt lymphoma cells.
AuthorsKelly GL, Milner AE, Baldwin GS, Bell AI, Rickinson AB
JournalProc Natl Acad Sci U S A
PubMed ID17001014
Epstein-Barr virus (EBV), a human herpesvirus, transforms B cell growth in vitro through expressing six virus-coded Epstein-Barr nuclear antigens (EBNAs) and two latent membrane proteins (LMPs). In many EBV-associated tumors, however, viral antigen expression is more restricted, and the aetiological role of the virus is unclear. For example, endemic Burkitt ... More
Migration of perilesional microglia after focal brain injury and modulation by CC chemokine receptor 5: an in situ time-lapse confocal imaging study.
AuthorsCarbonell WS, Murase S, Horwitz AF, Mandell JW
JournalJ Neurosci
PubMed ID16049180
Microglia rapidly become reactive in response to diverse stimuli and are thought to be prominent participants in the pathophysiology of both acute injury and chronic neurological diseases. However, mature microglial reactions to a focal lesion have not been characterized dynamically in adult vertebrate tissue. Here, we present a detailed analysis ... More