Signaling through MHC class II molecules blocks CD95-induced apoptosis.
AuthorsCatlett IM,Xie P,Hostager BS,Bishop GA
JournalJournal of immunology (Baltimore, Md. : 1950)
PubMed ID11342618
Glutamate and non-glutamate receptor mediated toxicity caused by oxygen and glucose deprivation in organotypic hippocampal cultures.
AuthorsNewell DW, Barth A, Papermaster V, Malouf AT
JournalJ Neurosci
PubMed ID7472521
In vitro ischemia models have utilized oxygen, or oxygen and glucose deprivation to simulate ischemic neuronal injury. Combined oxygen and glucose deprivation can induce neuronal damage which is in part mediated through NMDA receptors. Severe oxygen deprivation alone however can cause neuronal injury which is not NMDA mediated. We tested ... More
Identification and characterization of two subpopulations of Encephalitozoon intestinalis.
AuthorsHoffman RM, Marshall MM, Polchert DM, Jost BH
JournalAppl Environ Microbiol
PubMed ID12902292
Microsporidia are obligate intracellular protozoa that have been shown to be pathogenic to most living creatures. The development of in vitro cell culture propagation methods has provided researchers with large numbers of spores and facilitated the study of these organisms. Here, we describe heterogeneity within cell culture-propagated Encephalitozoon intestinalis suspensions. ... More
Autoantigens targeted in systemic lupus erythematosus are clustered in two populations of surface structures on apoptotic keratinocytes.
AuthorsCasciola-Rosen LA, Anhalt G, Rosen A
JournalJ Exp Med
PubMed ID7511686
'Systemic lupus erythematosus is a multisystem autoimmune disease in which the autoantibody response targets a variety of autoantigens of diverse subcellular location. We show here that these autoantigens are clustered in two distinct populations of blebs at the surface of apoptotic cells. The population of smaller blebs contains fragmented endoplasmic ... More
Caspase activation contributes to delayed death of heat-stressed striatal neurons.
AuthorsWhite MG, Emery M, Nonner D, Barrett JN
JournalJ Neurochem
PubMed ID14622126
'Hyperthermia can contribute to brain damage both during development and post-natally. We used rat embryonic striatal neurons in culture to study mechanisms underlying hyperthermia-induced neuronal death. Heat stress at 43 degrees C for 2 h produced no obvious signs of damage during the first 12 h after the stress, but ... More
Blockade of alpha 5 beta 1 integrins reverses the inhibitory effect of tenascin on chemotaxis of human monocytes and polymorphonuclear leukocytes through three-dimensional gels of extracellular matrix proteins.
AuthorsLoike JD, Cao L, Budhu S, Hoffman S, Silverstein SC
JournalJ Immunol
PubMed ID11390508
'Tenascin is an extracellular matrix protein found in adults in T cell-dependent areas of lymphoid tissues, sites of inflammation, and tumors. We report here that it inhibited chemotaxis of chemoattractant-stimulated human monocytes and chemoattractant-stimulated polymorphonuclear leukocytes (PMN) through three-dimensional gels composed of collagen I or Matrigel, and chemotaxis of leukotriene ... More
Detection of low copy numbers of HPV DNA by fluorescent in situ hybridization combined with confocal microscopy as an alternative to in situ polymerase chain reaction.
AuthorsLizard G, Chignol MC, Souchier C, Roignot P, Chardonnet Y, Schmitt D
JournalJ Virol Methods
PubMed ID9672129
'In genital lesions infected by human papillomavirus (HPV), histological criteria and HPV DNA typing are of prognostic value. Therefore, non-radioactive methods such as in situ hybridization are used extensively since they preserve the histological organization of the tissue, and allow the detection and characterization of HPV DNA. However, the sensitivity ... More
Ectopic expression of Drosophila ELAV and human HuD in Drosophila wing disc cells reveals functional distinctions and similarities.
AuthorsToba G, Qui J, Koushika SP, White K
JournalJ Cell Sci
PubMed ID12006625
'Drosophila ELAV and human HuD are two neuronal RNA binding proteins that show remarkable sequence homology, yet differ in their respective documented roles in post-transcriptional regulation. ELAV regulates neural-specific alternative splicing of specific transcripts, and HuD stabilizes specific mRNAs that are otherwise unstable due to AU-rich elements (AREs) in their ... More
Identification of an amino acid residue in multidrug resistance protein 1 critical for conferring resistance to anthracyclines.
AuthorsZhang DW, Cole SP, Deeley RG
JournalJ Biol Chem
PubMed ID11278596
'Murine multidrug resistance protein 1 (mrp1), unlike human MRP1, does not confer resistance to anthracyclines. Previously, we have shown that a human/murine hybrid protein containing amino acids 959-1187 of MRP1 can confer resistance to these drugs. We have now examined the functional characteristics of mutant proteins in which we have ... More
Flow cytometric methods for the analysis of human basophil surface antigens and viability.
AuthorsBochner BS, McKelvey AA, Schleimer RP, Hildreth JE, MacGlashan DW
JournalJ Immunol Methods
PubMed ID2607157
'Fluorescence and flow microfluorometric methods have been established for the detection and evaluation of IgE-bearing human leukocytes in various cell preparations including those where basophils are present at low percentages. Quantitative techniques for the determination of basophil purity, viability, and cell surface antigens including IgE are described. Use of these ... More
Cycloheximide-induced T-cell death is mediated by a Fas-associated death domain-dependent mechanism.
AuthorsTang D, Lahti JM, Grenet J, Kidd VJ
JournalJ Biol Chem
PubMed ID10066786
'Cycloheximide (CHX) can contribute to apoptotic processes, either in conjunction with another agent (e.g. tumor necrosis factor-alpha) or on its own. However, the basis of this CHX-induced apoptosis has not been clearly established. In this study, the molecular mechanisms of CHX-induced cell death were examined in two different human T-cell ... More
AuthorsChiang AS, Lin WY, Liu HP, Pszczolkowski MA, Fu TF, Chiu SL, Holbrook GL
JournalProc Natl Acad Sci U S A
PubMed ID11773617
'In vertebrates, the N-methyl-D-aspartate subtype of glutamate receptors (NMDAR) appears to play a role in neuronal development, synaptic plasticity, memory formation, and pituitary activity. However, functional NMDAR have not yet been characterized in insects. We have now demonstrated immunohistochemically glutamatergic nerve terminals in the corpora allata of an adult female ... More
The polo-box-dependent induction of ectopic septal structures by a mammalian polo kinase, plk, in Saccharomyces cerevisiae.
AuthorsLee KS, Song S, Erikson RL
JournalProc Natl Acad Sci U S A
PubMed ID10588710
'Members of the polo subfamily of protein kinases play pivotal roles in cell-cycle control and proliferation. In addition to a high degree of sequence similarity in the kinase domain, polo kinases contain a strikingly conserved motif termed "polo-box" in the noncatalytic C-terminal domain. We have previously shown that the mammalian ... More
Fluorescein-labeled tyramide strongly enhances the detection of low bromodeoxyuridine incorporation levels.
AuthorsVan Heusden J, de Jong P, Ramaekers F, Bruwiere H, Borgers M, Smets G
JournalJ Histochem Cytochem
PubMed ID9016320
'Immunocytochemical detection of bromodeoxyuridine (BrdU) labeling can be hampered by low BrdU incorporation levels. We describe here an amplification method for weak BrdU immunosignals. The tyramide signal amplification method based on catalyzed reporter deposition (CARD) uses fluorescein-labeled tyramide as a substrate for horseradish peroxidase. The enzyme catalyzes the formation of ... More
Multipotent stem cells from trabecular meshwork become phagocytic TM cells.
AuthorsDu Y, Roh DS, Mann MM, Funderburgh ML, Funderburgh JL, Schuman JS,
JournalInvest Ophthalmol Vis Sci
PubMed ID22297497
'To isolate and characterize stem cells from human trabecular meshwork (TM) and to investigate the potential of these stem cells to differentiate into TM cells. Human trabecular meshwork stem cells (TMSCs) were isolated as side population cells by fluorescence-activated cell sorting or isolated by clonal cultures. Passaged TMSCs were compared ... More
Flow cytometric detection of glutathione S-transferase isoenzymes by quantitative immunofluorescence under nonsaturating conditions.
AuthorsPifferi A, Filippini C, Capolongo L, Codegoni A, Balconi G, Ubezio P, Tagliabue G
JournalCytometry
PubMed ID7664624
'The glutathione (GSH)-glutathione S-transferase (GST) detoxification system is an important element in cellular defence against injurious agents and anticancer drugs. GST isoenzymes may represent biochemical markers of neoplastic transformation, and, possibly, drug resistance is associated with altered GST-isoenzyme levels. The ability to measure GST-isoenzymes in cell populations would be useful ... More
Targeting of the c-Abl tyrosine kinase to mitochondria in the necrotic cell death response to oxidative stress.
AuthorsKumar S, Bharti A, Mishra NC, Raina D, Kharbanda S, Saxena S, Kufe D
JournalJ Biol Chem
PubMed ID11350980
'The ubiquitously expressed c-Abl tyrosine kinase is activated in the response of cells to genotoxic and oxidative stress. The present study demonstrates that reactive oxygen species (ROS) induce targeting of c-Abl to mitochondria. We show that ROS-induced localization of c-Abl to mitochondria is dependent on activation of protein kinase C ... More
Selective cell targeting with light-absorbing microparticles and nanoparticles.
AuthorsPitsillides CM, Joe EK, Wei X, Anderson RR, Lin CP
JournalBiophys J
PubMed ID12770906
'We describe a new method for selective cell targeting based on the use of light-absorbing microparticles and nanoparticles that are heated by short laser pulses to create highly localized cell damage. The method is closely related to chromophore-assisted laser inactivation and photodynamic therapy, but is driven solely by light absorption, ... More
KIFC3, a microtubule minus end-directed motor for the apical transport of annexin XIIIb-associated Triton-insoluble membranes.
AuthorsNoda Y, Okada Y, Saito N, Setou M, Xu Y, Zhang Z, Hirokawa N
JournalJ Cell Biol
PubMed ID11581287
'We have identified and characterized a COOH-terminal motor domain-type kinesin superfamily protein (KIFC), KIFC3, in the kidney. KIFC3 is a minus end-directed microtubule motor protein, therefore it accumulates in regions where minus ends of microtubules assemble. In polarized epithelial cells, KIFC3 is localized on membrane organelles immediately beneath the apical ... More
Zygotic expression of the pebble locus is required for cytokinesis during the postblastoderm mitoses of Drosophila.
AuthorsHime G, Saint R
JournalDevelopment
PubMed ID1576956
'Mutations at the pebble locus of Drosophila melanogaster result in embryonic lethality. Examination of homozygous mutant embryos at the end of embryogenesis revealed the presence of fewer and larger cells which contained enlarged nuclei. Characterization of the embryonic cell cycles using DAPI, propidium iodide, anti-tubulin and anti-spectrin staining showed that ... More
The ubiquitin-like protein FAT10 forms covalent conjugates and induces apoptosis.
AuthorsRaasi S, Schmidtke G, Groettrup M
JournalJ Biol Chem
PubMed ID11445583
'FAT10 is a ubiquitin-like protein that is encoded in the major histocompatibility complex class I locus and is synergistically inducible with interferon-gamma and tumor necrosis factor alpha. The molecule consists of two ubiquitin-like domains in tandem arrangement and bears a conserved diglycine motif at its carboxyl terminus commonly used in ... More
Antiangiogenesis signals by endostatin.
AuthorsShichiri M, Hirata Y
JournalFASEB J
PubMed ID11292666
'Endostatin is a potent endogenous angiogenesis inhibitor that induces regression of tumors in mice. Neither an extracellular receptor for endostatin nor intracellular signals that result in the regression of tumor vascular beds have been identified. We demonstrate that endostatin, but not angiostatin, at comparable concentrations to those used in in ... More
Alexa dyes, a series of new fluorescent dyes that yield exceptionally bright, photostable conjugates.
AuthorsPanchuk-Voloshina N, Haugland RP, Bishop-Stewart J, Bhalgat MK, Millard PJ, Mao F, Leung WY, Haugland RP
JournalJ Histochem Cytochem
PubMed ID10449539
'Alexa 350, Alexa 430, Alexa 488, Alexa 532, Alexa 546, Alexa 568, and Alexa 594 dyes are a new series of fluorescent dyes with emission/excitation spectra similar to those of AMCA, Lucifer Yellow, fluorescein, rhodamine 6G, tetramethylrhodamine or Cy3, lissamine rhodamine B, and Texas Red, respectively (the numbers in the ... More
Distinct Intracellular Signaling in Tumor Necrosis Factor-related Apoptosis-inducing Ligand- and CD95 Ligand-mediated Apoptosis.
Authors Velthuis Jurjen H L; Rouschop Kasper M A; De Bont Hans J G M; Mulder Gerard J; Nagelkerke J Fred;
JournalJ Biol Chem
PubMed ID11980895
'Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potent inducer of apoptosis in tumor cells but not in healthy cells. Similar to CD95 ligand (CD95L), TRAIL signaling requires ligand-receptor interaction; the downstream signaling molecules, such as Fas-associated death domain and caspase-8, also seem similar. Using cells stably expressing TRAIL and ... More
Loss of PINK1 function promotes mitophagy through effects on oxidative stress and mitochondrial fission.
AuthorsDagda RK, Cherra SJ, Kulich SM, Tandon A, Park D, Chu CT,
JournalJ Biol Chem
PubMed ID19279012
'Mitochondrial dysregulation is strongly implicated in Parkinson disease. Mutations in PTEN-induced kinase 1 (PINK1) are associated with familial parkinsonism and neuropsychiatric disorders. Although overexpressed PINK1 is neuroprotective, less is known about neuronal responses to loss of PINK1 function. We found that stable knockdown of PINK1 induced mitochondrial fragmentation and autophagy ... More
Visualization of the compartmentalization of glutathione and protein-glutathione mixed disulfides in cultured cells.
AuthorsSöderdahl T, Enoksson M, Lundberg M, Holmgren A, Ottersen OP, Orrenius S, Bolcsfoldi G, Cotgreave IA
JournalFASEB J
PubMed ID12475911
'Fluorescence microscopy of A549 cells stained with a glutathione (L-gamma-glutamyl-L-cysteinylglycine, GSH)-specific polyclonal antibody displayed uniform staining of the peri-nuclear cytosol, with the nuclear region apparently lacking GSH staining. This discontinuous staining was confirmed in other cell types and also corroborated in A549 cells stained with the thiol-reactive dye mercury orange. ... More
Candida albicans phospholipomannan promotes survival of phagocytosed yeasts through modulation of bad phosphorylation and macrophage apoptosis.
AuthorsIbata-Ombetta S, Idziorek T, Trinel PA, Poulain D, Jouault T
JournalJ Biol Chem
PubMed ID12551950
'The surface of the pathogenic yeast Candida albicans is coated with phospholipomannan (PLM), a phylogenetically unique glycolipid composed of beta-1,2-oligomannosides and phytoceramide. This study compared the specific contribution of PLM to the modulation of signaling pathways linked to the survival of C. albicans in macrophages in contrast to Saccharomyces cerevisiae. ... More
Bcl-2 expression in synovial fibroblasts is essential for maintaining mitochondrial homeostasis and cell viability.
'The regulation of proliferation and cell death is vital for homeostasis, but the mechanism that coordinately balances these events in rheumatoid arthritis (RA) remains largely unknown. In RA, the synovial lining thickens in part through increased proliferation and/or decreased synovial fibroblast cell death. Here we demonstrate that the anti-apoptotic protein, ... More
Glycoprotein IIb/IIIa antagonists induce apoptosis in rat cardiomyocytes by caspase-3 activation.
AuthorsAdderley SR, Fitzgerald DJ
JournalJ Biol Chem
PubMed ID10681563
'The platelet integrin glycoprotein (GP) IIb/IIIa, which mediates platelet aggregation, has been the target for novel antiplatelet agents, the GPIIb/IIIa antagonists. Several GPIIb/IIIa antagonists have been developed based on the peptide RGDS present in adhesion proteins, including the principle ligand fibrinogen. The apoptosis enzyme, procaspase-3, contains an RGD-recognition sequence and ... More
A seed for Alzheimer amyloid in the brain.
AuthorsHayashi H, Kimura N, Yamaguchi H, Hasegawa K, Yokoseki T, Shibata M, Yamamoto N, Michikawa M, Yoshikawa Y, Terao K, Matsuzaki K, Lemere CA, Selkoe DJ, Naiki H, Yanagisawa K
JournalJ Neurosci
PubMed ID15152051
'A fundamental question about the early pathogenesis of Alzheimer''s disease (AD) concerns how toxic aggregates of amyloid beta protein (Abeta) are formed from its nontoxic soluble form. We hypothesized previously that GM1 ganglioside-bound Abeta (GAbeta) is involved in the process. We now examined this possibility using a novel monoclonal antibody ... More
Simultaneous flow cytometric detection of cellular c-myc protein, incorporated bromodeoxyuridine, and DNA.
AuthorsRosette CD, DeTeresa PS, Pallavicini MG
JournalCytometry
PubMed ID2111761
'We describe a multivariate flow cytometric technique for simultaneous analysis of specific nuclear protein, bromodeoxyuridine (BrdUrd) incorporated into DNA and DNA content in single cells in suspension. The procedure involves fixation of BrdUrd-exposed cells with paraformaldehyde, heat denaturation of cellular DNA, followed by sequential immunochemical reactions to label incorporated BrdUrd ... More
'The generation of micronuclei is a reflection of DNA damage, defective mitosis, and loss of genetic material. The involvement of the MAPK pathway in mediating v-ras-induced micronuclei in NIH 3T3 cells was examined by inhibiting MAPK activation. Conversely, the MAPK pathway was constitutively activated by infecting cells with a v-mos ... More
Sensitive multicolor fluorescence in situ hybridization using catalyzed reporter deposition (CARD) amplification.
AuthorsSpeel EJ, Ramaekers FC, Hopman AH
JournalJ Histochem Cytochem
PubMed ID9313806
'We describe the simultaneous localization of DNA sequences in cell and chromosome preparations by means of differently fluorochrome-labeled (AMCA, FITC, TRITC) tyramides using the catalyzed reporter deposition (CARD) procedure. For this purpose, repeated as well as single-copy DNA probes were labeled with biotin, digoxigenin, and FITC, hybridized, and visualized with ... More
Roles of Rho-associated kinase in cytokinesis; mutations in Rho-associated kinase phosphorylation sites impair cytokinetic segregation of glial filaments.
AuthorsYasui Y, Amano M, Nagata K, Inagaki N, Nakamura H, Saya H, Kaibuchi K, Inagaki M
JournalJ Cell Biol
PubMed ID9832553
'Rho-associated kinase (Rho-kinase), which is activated by the small GTPase Rho, regulates formation of stress fibers and focal adhesions, myosin fiber organization, and neurite retraction through the phosphorylation of cytoskeletal proteins, including myosin light chain, the ERM family proteins (ezrin, radixin, and moesin) and adducin. Rho-kinase was found to phosphorylate ... More
Bcl-2 controls caspase activation following a p53-dependent cyclin D1-induced death signal.
AuthorsPratt MA, Niu MY
JournalJ Biol Chem
PubMed ID12480939
'MCF-7 and ZR-75 breast cancer cells infected with an adenovirus constitutively expressing high levels of cyclin D1 demonstrated widespread mitochondrial translocation of Bax and cytochrome c release that was approximately doubled after the addition of all-trans retinoic acid (RA) or Bcl-2 antisense oligonucleotide. By comparison, the percentage of cells in ... More
High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones.
'Cosmid clones containing human DNA inserts have been mapped on chromosome 11 by fluorescence in situ hybridization under conditions that suppress signal from repetitive DNA sequences. Thirteen known genes, one chromosome 11-specific DNA repeat, and 36 random clones were analyzed. High-resolution mapping was facilitated by using digital imaging microscopy and ... More
Adenovirus-mediated overexpression of catalase in the cytosolic or mitochondrial compartment protects against cytochrome P450 2E1-dependent toxicity in HepG2 cells.
AuthorsBai J, Cederbaum AI
JournalJ Biol Chem
PubMed ID11071897
'Cytochrome P450 2E1 (CYP2E1) is an effective producer of reactive oxygen species such as superoxide radical and hydrogen peroxide, which may contribute to the development of alcohol liver disease or cytotoxicity. To investigate the protective role of catalase against CYP2E1-dependent cytotoxicity, E47 cells, a transfected HepG2 cell line overexpressing CYP2E1, ... More
Affinity modulation of platelet integrin alphaIIbbeta3 by beta3-endonexin, a selective binding partner of the beta3 integrin cytoplasmic tail.
AuthorsKashiwagi H, Schwartz MA, Eigenthaler M, Davis KA, Ginsberg MH, Shattil SJ
JournalJ Cell Biol
PubMed ID9182673
'Platelet agonists increase the affinity state of integrin alphaIIbbeta3, a prerequisite for fibrinogen binding and platelet aggregation. This process may be triggered by a regulatory molecule(s) that binds to the integrin cytoplasmic tails, causing a structural change in the receptor. beta3-Endonexin is a novel 111-amino acid protein that binds selectively ... More
Nuclear localization of L-type glutaminase in mammalian brain.
AuthorsOlalla L, Gutiérrez A, Campos JA, Khan ZU, Alonso FJ, Segura JA, Márquez J, Aledo JC
JournalJ Biol Chem
PubMed ID12163477
'In mammals, there are two different genes encoding for glutaminase isoforms, named liver (LGA) and kidney (KGA) types. LGA has long been believed to be present only in liver mitochondria from adult animals. However, we have recently reported the presence of LGA mRNA in human brain. We now describe the ... More
Neogenesis of cerebellar Purkinje neurons from gene-marked bone marrow cells in vivo.
AuthorsPriller J, Persons DA, Klett FF, Kempermann G, Kreutzberg GW, Dirnagl U
JournalJ Cell Biol
PubMed ID11724815
'The versatility of stem cells has only recently been fully recognized. There is evidence that upon adoptive bone marrow (BM) transplantation (BMT), donor-derived cells can give rise to neuronal phenotypes in the brains of recipient mice. Yet only few cells with the characteristic shape of neurons were detected 1-6 mo ... More
Improved method for computing potential doubling time from flow cytometric data.
AuthorsWhite RA, Terry NH, Meistrich ML, Calkins DP
JournalCytometry
PubMed ID2318085
'Relative movement methods use the timed progression of the mean fluorescence of cells which have been labeled with monoclonal antibodies against bromodeoxyuridine and displayed with bivariate flow cytometry according to DNA and label content to compute duration of DNA synthesis, TS. The relative movement is the difference of the mean ... More
Beta-catenin-induced melanoma growth requires the downstream target Microphthalmia-associated transcription factor.
AuthorsWidlund HR, Horstmann MA, Price ER, Cui J, Lessnick SL, Wu M, He X, Fisher DE
JournalJ Cell Biol
PubMed ID12235125
'The transcription factor Microphthalmia-associated transcription factor (MITF) is a lineage-determination factor, which modulates melanocyte differentiation and pigmentation. MITF was recently shown to reside downstream of the canonical Wnt pathway during melanocyte differentiation from pluripotent neural crest cells in zebrafish as well as in mammalian melanocyte lineage cells. Although expression of ... More
Cytochrome c-mediated apoptosis in cells lacking mitochondrial DNA. Signaling pathway involving release and caspase 3 activation is conserved.
AuthorsJiang S, Cai J, Wallace DC, Jones DP
JournalJ Biol Chem
PubMed ID10514472
'Mitochondria serve as a pivotal component of the apoptotic cell death machinery. However, cells that lack mitochondrial DNA (rho(0) cells) retain apparently normal apoptotic signaling. In the present study, we examined mitochondrial mechanisms of apoptosis in rho(0) osteosarcoma cells treated with staurosporine. Immunohistochemistry revealed that rho(0) cells maintained a normal ... More
Activation of extracellular-regulated kinase pathways in ovarian granulosa cells by the novel growth factor type 1 follicle-stimulating hormone receptor. Role in hormone signaling and cell proliferation.
'Follicle-stimulating hormone (FSH) regulated growth and function of the ovarian follicle was previously thought to be mediated solely through activation of G(s)-coupled receptors. In this study, we show for the first time that this function is predominantly mediated through the alternatively spliced and novel growth factor type 1 receptor (oFSH-R3) ... More
Caffeine overcomes a restriction point associated with DNA replication, but does not accelerate mitosis.
AuthorsDownes CS, Musk SR, Watson JV, Johnson RT
JournalJ Cell Biol
PubMed ID2161852
'Mitotic chromosome condensation is normally dependent on the previous completion of replication. Caffeine spectacularly deranges cell cycle controls after DNA polymerase inhibition or DNA damage; it induces the condensation, in cells that have not completed replication, of fragmented nuclear structures, analogous to the S-phase prematurely condensed chromosomes seen when replicating ... More
Cadherins mediate intercellular mechanical signaling in fibroblasts by activation of stretch-sensitive calcium-permeable channels.
AuthorsKo KS, Arora PD, McCulloch CA
JournalJ Biol Chem
PubMed ID11466312
'Cells in mechanically active environments form extensive, cadherin-mediated intercellular junctions that are important in tissue remodeling and differentiation. Currently, it is unknown whether adherens junctions in connective tissue fibroblasts transmit mechanical signals and coordinate multicellular adaptations to physical forces. We hypothesized that cadherins mediate intercellular mechanotransduction by activating calcium-permeable, stretch-sensitive ... More
A flow cytometric method for measuring lymphocyte proliferation directly from tissue culture plates using Ki-67 and propidium iodide.
AuthorsPalutke M, KuKuruga D, Tabaczka P
JournalJ Immunol Methods
PubMed ID2960745
'A new method for measuring lymphocyte proliferation in response to mitogens and allogeneic cells without using radiolabelling is described. It utilizes flow cytometry and the monoclonal antibody, Ki-67, which detects a nuclear proliferation antigen. The entire test is performed in standard, 96-well tissue culture plates. Stable, clean nuclear suspensions rather ... More
Quantification of Bax and Bcl2 in polymorphonuclear leukocytes from haemodialysis patients: relation to hydrogen peroxide.
'BACKGROUND: Bax and Bcl2 are two apoptosis-related molecules that play an important role in determining cell fate following oxidative injury. In the present study, we explored the relation of hydrogen peroxide (H2O2) generation by polymorphonuclear cells (PMNs) to the cytosolic expression of Bax and Bcl2 proteins and apoptosis in haemodialysis ... More
The transcription factor AP-1 is required for EGF-induced activation of rho-like GTPases, cytoskeletal rearrangements, motility, and in vitro invasion of A431 cells.
AuthorsMalliri A, Symons M, Hennigan RF, Hurlstone AF, Lamb RF, Wheeler T, Ozanne BW
JournalJ Cell Biol
PubMed ID9817764
'Human squamous cell carcinomas (SCC) frequently express elevated levels of epidermal growth factor receptor (EGFR). EGFR overexpression in SCC-derived cell lines correlates with their ability to invade in an in vitro invasion assay in response to EGF, whereas benign epidermal cells, which express low levels of EGFR, do not invade. ... More
An improved method for the detection of cell surface antigens in samples of low viability using flow cytometry.
'A high non-specific background fluorescence signal was observed when cell surface antigen analysis was carried out using flow cytometry on a cell sample which contained a high proportion of dead and dying cells. To overcome this problem it was necessary to analyse the cells in three stages. First the intact ... More
Deficiencies in collagen phagocytosis by human fibroblasts in vitro: a mechanism for fibrosis?
AuthorsMcCulloch CA, Knowles GC
JournalJ Cell Physiol
PubMed ID8491787
'Degradation of collagen by fibroblast phagocytosis is an important pathway for physiological remodelling of soft connective tissues. Perturbations of this pathway may provide a mechanism for the development of fibrotic lesions. As collagen phagocytosis may be regulated by either a change of the proportions or the activity of phagocytic cells, ... More
Immunoseparation and immunodetection of nucleic acids labeled with halogenated nucleotides.
AuthorsHaider SR, Juan G, Traganos F, Darzynkiewicz Z
JournalExp Cell Res
PubMed ID9260920
'A novel methodology for labeling, isolation, and detection of nucleic acids is described. Nucleic acid isolation is based on in vivo or in vitro incorporation of BrU or BrdU to either RNA or DNA, respectively, followed by immunoprecipitation of the labeled nucleic acid utilizing anti-BrdU MoAb, which crossreacts with BrU, ... More
Redistribution and dysfunction of integrins in cultured renal epithelial cells exposed to oxidative stress.
AuthorsGailit J, Colflesh D, Rabiner I, Simone J, Goligorsky MS
JournalAm J Physiol
PubMed ID8430825
'Tubular obstruction by detached renal tubular epithelial cells is a major cause of oliguria in acute renal failure. Viable renal tubular cells can be recovered from urine of patients with acute tubular necrosis, suggesting a possible defect in cell adhesion to the basement membrane. To study this process of epithelial ... 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
Apical localization of wingless transcripts is required for wingless signaling.
AuthorsSimmonds AJ, dosSantos G, Livne-Bar I, Krause HM
JournalCell
PubMed ID11336670
'Many developing and adult tissues are comprised of polarized epithelia. Proteins that are asymmetrically distributed in these cells are thought to be localized by protein trafficking. Here we show that the distribution and function of the signaling protein Wingless is predetermined by the subcellular localization of its mRNA. High-resolution in ... More
Bcl-xL regulates the membrane potential and volume homeostasis of mitochondria.
'Mitochondrial physiology is disrupted in either apoptosis or necrosis. Here, we report that a wide variety of apoptotic and necrotic stimuli induce progressive mitochondrial swelling and outer mitochondrial membrane rupture. Discontinuity of the outer mitochondrial membrane results in cytochrome c redistribution from the intermembrane space to the cytosol followed by ... More
The GTP-binding protein Rho1p is required for cell cycle progression and polarization of the yeast cell.
AuthorsDrgonová J, Drgon T, Roh DH, Cabib E
JournalJ Cell Biol
PubMed ID10427091
'Previous work showed that the GTP-binding protein Rho1p is required in the yeast, Saccharomyces cerevisiae, for activation of protein kinase C (Pkc1p) and for activity and regulation of beta(1-->3)glucan synthase. Here we demonstrate a hitherto unknown function of Rho1p required for cell cycle progression and cell polarization. Cells of mutant ... More
Importance of MEK in neutrophil microbicidal responsiveness.
'Exposure of neutrophils to inflammatory stimuli such as the chemoattractant FMLP leads to activation of responses including cell motility, the oxidative burst, and secretion of proteolytic enzymes. A signaling cascade involving sequential activation of Raf-1, mitogen-activated protein kinase (MEK), and extracellular signal regulated kinase (ERK) is also rapidly activated after ... More
The drosophila protein asp is involved in microtubule organization during spindle formation and cytokinesis.
AuthorsWakefield JG, Bonaccorsi S, Gatti M
JournalJ Cell Biol
PubMed ID11352927
'Abnormal spindle (Asp) is a 220-kD microtubule-associated protein from Drosophila that has been suggested to be involved in microtubule nucleation from the centrosome. Here, we show that Asp is enriched at the poles of meiotic and mitotic spindles and localizes to the minus ends of central spindle microtubules. Localization to ... More
An analysis of changes in the expression of cyclins A and B1 by the cell array system during the cell cycle: comparison between cell synchronization methods.
AuthorsTakita M, Furuya T, Sugita T, Kawauchi S, Oga A, Hirano T, Tsunoda S, Sasaki K
JournalCytometry A
PubMed ID12938185
'BACKGROUND: A novel high-throughput analysis, cell array system, was developed for an extensive study of the expression of genes and/or the degradation of gene products at the cellular level. To exemplify the usefulness of this system, we showed the changes in the expression level of cyclin A and B1 during ... More
Desmosomal proteins, including desmoglein 3, serve as novel negative markers for epidermal stem cell-containing population of keratinocytes.
AuthorsWan H, Stone MG, Simpson C, Reynolds LE, Marshall JF, Hart IR, Hodivala-Dilke KM, Eady RA
JournalJ Cell Sci
PubMed ID12953062
'No single method has been universally adopted for identifying and isolating epidermal stem/progenitor cells, and the emergence of new markers of stem cell populations is worth exploring. Here we report, for the first time, that clusters of basal keratinocytes at the tips of the rete ridges in human palm, previously ... More
Temperature-dependent arrest of neutrophil apoptosis. Failure of Bax insertion into mitochondria at 15 degrees C prevents the release of cytochrome c.
AuthorsPryde JG, Walker A, Rossi AG, Hannah S, Haslett C
JournalJ Biol Chem
PubMed ID10896657
'Apoptosis is essential for the resolution of neutrophilic inflammation. To define the mechanisms triggering the execution phase of apoptosis we developed and utilized a model in which culture of human neutrophils at 15 degrees C for 20 h arrested apoptosis and subsequent warming to 37 degrees C triggered a synchronous ... More
v-Abl protein-tyrosine kinase up-regulates p21WAF-1 in cell cycle arrested and proliferating myeloid cells.
AuthorsKhanna SJ, Brown R, Whetton AD, Ball KL, Dive C
JournalJ Biol Chem
PubMed ID11098052
'v-Abl protein-tyrosine kinase (PTK) promotes cell survival without cell proliferation in interleukin (IL)-3-deprived IC.DP premast cells (1). We now show that in these conditions v-Abl PTK transcriptionally up-regulated the cyclin-dependent kinase inhibitor (CDKI) p21(WAF-1) and inhibited CDK2 and CDK4. When readdition of IL-3 stimulated cell proliferation, p21(WAF-1) was inactivated as ... More
Zinc depletion reduced Egr-1 and HNF-3beta expression and apolipoprotein A-I promoter activity in Hep G2 cells.
AuthorsCui L, Schoene NW, Zhu L, Fanzo JC, Alshatwi A, Lei KY
JournalAm J Physiol Cell Physiol
PubMed ID12107072
'We examined the influence of zinc status on expression of certain transcription factors involved in regulation of apolipoprotein A-I (apoAI) expression in human hepatoblastoma Hep G2 cells. A low zinc basal medium (zinc deficient, ZD) consisting of DMEM and 10% Chelex100-treated fetal bovine serum was used to deplete cellular zinc ... More
Application of differential inflammatory cell count as a tool to monitor udder health.
AuthorsPillai SR, Kunze E, Sordillo LM, Jayarao BM
JournalJ Dairy Sci
PubMed ID11417700
'A flow cytometric technique called differential inflammatory cell count was standardized by staining bovine peripheral blood leukocytes with a combination of DNA binding dyes SYBR green 1 and propidium iodide in water. Leukocytes were also stained with propidium iodide in detergent to determine total cell count. Differential inflammatory cell count ... More
A novel cytotoxicity screening assay using a multiwell fluorescence scanner.
AuthorsNieminen AL, Gores GJ, Bond JM, Imberti R, Herman B, Lemasters JJ
JournalToxicol Appl Pharmacol
PubMed ID1641848
'A new assay using a multiwell fluorescence scanner was developed for screening cytotoxicity to cells cultured in 96-well microtiter plates. The assay is based on binding of propidium iodide to nuclei of cells whose plasma membranes have become permeable due to cell death. Fluorescence of propidium iodide measured with a ... More
An antisense oligonucleotide to 1-cys peroxiredoxin causes lipid peroxidation and apoptosis in lung epithelial cells.
AuthorsPak JH, Manevich Y, Kim HS, Feinstein SI, Fisher AB
JournalJ Biol Chem
PubMed ID12372839
'1-cys peroxiredoxin (1-cysPrx), a member of the peroxiredoxin superfamily, reduces phospholipid hydroperoxides as well as organic peroxides and H(2)O(2). To determine the physiological function(s) of 1-cysPrx, we have used an antisense strategy to suppress endogenous 1-cysPrx in L2 cells, a rat lung epithelial cell line. A 25-base antisense morpholino oligonucleotide ... More
Evaluation of cryopreserved stallion semen from Tori and Estonian breeds using CASA and flow cytometry.
AuthorsKavak A, Johannisson A, Lundeheim N, Rodriguez-Martinez H, Aidnik M, Einarsson S
JournalAnim Reprod Sci
PubMed ID12586493
'Methods to evaluate the quality of frozen-thawed stallion semen are still needed, particularly those considering the sperm function. The present study evaluated sperm motility, membrane and acrosome integrity and the capacitation status of frozen-thawed spermatozoa from seven Tori and six Estonian breed stallions by way of computer assisted sperm analysis ... More
Ethylene modulates gene expression in cells of the marine sponge Suberites domuncula and reduces the degree of apoptosis.
AuthorsKrasko A, Schröder HC, Perovic S, Steffen R, Kruse M, Reichert W, Müller IM, Müller WE
JournalJ Biol Chem
PubMed ID10531355
'Sponges (phylum Porifera) live in an aqueous milieu that contains dissolved organic carbon. This is degraded photochemically by ultraviolet radiation to alkenes, particularly to ethylene. This study demonstrates that sponge cells (here the demosponge Suberites domuncula has been used), which have assembled to primmorphs, react to 5 microM ethylene with ... More
Connexin26 is responsible for anionic molecule permeability in the cochlea for intercellular signalling and metabolic communications.
AuthorsZhao HB
JournalEur J Neurosci
PubMed ID15869481
'Abstract A gap junction is composed of two hemichannels and possesses a relatively large pore size ( approximately 10-15 A), allowing passage of ions and molecules up to 1 kDa. Here, we report that connexin hemichannels and gap junctions in the guinea pig cochlea had significant charge selectivity among permeating ... More
Comparison of Hoechst 33342 and propidium iodide as fluorescent markers for sperm fusion with hamster oocytes.
AuthorsGreen DP
JournalJ Reprod Fertil
PubMed ID1285064
'Hamster oocytes were loaded with the DNA dyes Hoechst 33342 or propidium iodide. Oocytes incubated in 10 mumol Hoechst 333421(-1) showed intracellular fluorescence within 10-20 s of exposure, as did hamster and guinea-pig spermatozoa. Impaled oocytes to which acrosome-intact hamster spermatozoa were bound before injection of Hoechst 33342 showed dye ... More
Flow cytometric analysis of mammalian glial cultures treated with methotrexate.
AuthorsSerrano EE, Schimke RT
JournalGlia
PubMed ID2148556
'Methotrexate (MTX) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase (DHFR). MTX treatment of cultured cell lines leads to the emergence of resistant cell populations. Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of DHFR can be caused by ... More
Diazoxide triggers cardioprotection against apoptosis induced by oxidative stress.
AuthorsIchinose M, Yonemochi H, Sato T, Saikawa T
JournalAm J Physiol Heart Circ Physiol
PubMed ID12623783
'Although mitochondrial ATP-sensitive potassium (mitoK(ATP)) channels have been reported to reduce the extent of apoptosis, the critical timing of mitoK(ATP) channel opening required to protect myocytes against apoptosis remains unclear. In the present study, we examined whether the mitoK(ATP) channel serves as a trigger of cardioprotection against apoptosis induced by ... 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
Isolation and characterization of primordial follicles from fresh and cryopreserved human ovarian tissue.
AuthorsOktay K, Nugent D, Newton H, Salha O, Chatterjee P, Gosden RG
JournalFertil Steril
PubMed ID9091334
'OBJECTIVE: To develop an efficient isolation technique for human primordial follicles. DESIGN: Prospective, experimental study of ovarian biopsies collected from healthy women undergoing elective cesarean section. Ovarian blocks either were fixed for histology and follicle counting or partially disaggregated with type 1A collagenase before or after cryopreservation. After partial disaggregation, ... 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
Use of methacrylate De-embedding protocols for in situ hybridization on semithin plastic sections with multiple detection strategies.
AuthorsWarren KC, Coyne KJ, Waite JH, Cary SC
JournalJ Histochem Cytochem
PubMed ID9446821
'New plastic resins are gradually replacing traditional paraffin-embedding in situ hybridization (ISH) strategies. One unique resin that has not been fully investigated or exploited with respect to light microscopic ISH is a methacrylate mixture. The advantage of this resin is its ability to be removed from tissues postsectioning, dramatically increasing ... More
Extended application of flow microfluorometry by means of dual laser excitation.
AuthorsStöhr M, Eipel H, Goerttler K, Vogt-Schaden M
JournalHistochemistry
PubMed ID870462
'A dual laser beam excitation device for flow analysis of biological particles has been developed. The aid of this arrangement is to increase the range of fluorescent agents employed so far in quantitative and qualitative cytochemistry. Combining an argon ion and a helium-cadmium laser two color fluorescence measurements were performed ... 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
Hidden effects of cryopreservation on quality of human spermatozoa.
AuthorsGlander HJ, Schaller J
JournalCell Tissue Bank
PubMed ID15256959
'The effects of cryopreservation on two characteristics of human spermatozoa were investigated: the early phases of disturbed plasma membrane function and the activity of enzymes in intact spermatozoa. The membrane function was detected by means of the calcium-dependent binding of fluorescein isothiocyanate (FITC)-conjugated Annexin V to sperm plasma membranes. Annexin ... More
Modulation of TGF-beta type 1 receptor: flow cytometric detection with biotinylated TGF-beta.
AuthorsNewman W, Beall LD, Bertolini DR, Cone JL
JournalJ Cell Physiol
PubMed ID2550480
'Transforming growth factor beta type 1 (TGF-beta 1) was reacted with NHS-biotin to yield a derivative of TGF-beta 1 which was biotinylated on lysine residues. The biotinylated form of TGF-beta 1 was separated from the unreacted material by reverse phase chromatography. In three separate bioassays, the derivatized peptide was as ... 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
The NF-kappa B cascade is important in Bcl-xL expression and for the anti-apoptotic effects of the CD28 receptor in primary human CD4+ lymphocytes.
AuthorsKhoshnan A, Tindell C, Laux I, Bae D, Bennett B, Nel AE
JournalJ Immunol
PubMed ID10925251
'We explored the role of the NF-kappa B pathway in the survival of primary human CD4+ T lymphocytes during CD28 costimulation. Transduction of proliferating CD4+ T cells with a tetracycline-regulated retrovirus encoding for a dominant-interfering, degradation-resistant I-kappaBalpha (inhibitor of kappa B alpha factor) mutant induced apoptosis. Using DNA arrays, we ... More
Ionizing radiation down-regulates p53 protein in primary Egr-1-/- mouse embryonic fibroblast cells causing enhanced resistance to apoptosis.
AuthorsDas A, Chendil D, Dey S, Mohiuddin M, Mohiuddin M, Milbrandt J, Rangnekar VM, Ahmed MM
JournalJ Biol Chem
PubMed ID11035041
'In this study, we sought to investigate the mechanism of the proapoptotic function of Egr-1 in relation to p53 status in normal isogenic cell backgrounds by using primary MEF cells established from homozygous (Egr-1(-/-)) and heterozygous (Egr-1(+/-)) Egr-1 knock-out mice. Ionizing radiation caused significantly enhanced apoptosis in Egr-1(+/-) cells (22.8%; ... More
Fluorescence detection of 8-oxoguanine in nuclear and mitochondrial DNA of cultured cells using a recombinant Fab and confocal scanning laser microscopy.
'The presence of 8-oxoguanine (8-oxoG) in DNA is considered a marker of oxidative stress and DNA damage. We describe a multifluorescence technique to detect the localization of 8-oxoG in both nuclear and mitochondrial DNA using a mouse recombinant Fab 166. The Fab was generated by repertoire cloning and combinatorial phage ... 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
Differential effects of deuterium oxide on the fluorescence lifetimes and intensities of dyes with different modes of binding to DNA.
AuthorsSailer BL, Nastasi AJ, Valdez JG, Steinkamp JA, Crissman HA
JournalJ Histochem Cytochem
PubMed ID9016307
'Deuterium oxide (D2O) increases both the fluorescence lifetime and the fluorescence intensity of the intercalating dyes propidium iodide (PI) and ethidium bromide (EB) when bound to nucleic acid structures. We have used spectroscopic analysis coupled with conventional and phase-sensitive flow cytometry to compare the alterations in intensity and lifetime of ... More
A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry.
AuthorsNicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C
JournalJ Immunol Methods
PubMed ID1710634
'Corticosteroids, calcium ionophores and anti-CD3 monoclonal antibodies kill mouse thymocytes incubated in vitro. Cell death is preceded by extensive DNA fragmentation into oligonucleosomal subunits. This type of cell death (apoptosis), which physiologically occurs in the intrathymic process of immune cell selection, is usually evaluated by either electrophoretic or colorimetric methods ... 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
Antimicrobial action of rabbit leukocyte CAP18(106-137).
AuthorsMason DJ, Dybowski R, Larrick JW, Gant VA
JournalAntimicrob Agents Chemother
PubMed ID9056004
'CAP18 is a cationic antimicrobial protein originally isolated from rabbit neutrophils, of which a 32-mer sequence from its C-terminal and (CAP18(106-137)) has been found to be the most active. The bactericidal action of this peptide has been characterized by conventional culture techniques and flow cytometry. Cultures of Escherichia coli NCTC10418 ... More
In vitro model of the outer blood-retina barrier.
AuthorsSteuer H, Jaworski A, Stoll D, Schlosshauer B
JournalBrain Res Brain Res Protoc
PubMed ID15063838
'The outer blood-retina barrier (BRB) is formed by the retinal pigment epithelium (rpe) and functions similarly to the blood-brain barrier (BBB). In contrast to the BBB, which is composed of a myriad of capillaries, the rpe can in principle be prepared as an intact planar tissue sheet without disruption of ... More
Flow cytometric discrimination of mitotic cells: resolution of M, as well as G1, S, and G2 phase nuclei with mithramycin, propidium iodide, and ethidium bromide after fixation with formaldehyde.
AuthorsLarsen JK, Munch-Petersen B, Christiansen J, Jørgensen K
JournalCytometry
PubMed ID2419056
'Cells in mitosis can be flow cytometrically discriminated from G1, S, and G2 cells by analysis of a nuclear suspension prepared with nonionic detergent, fixed with formaldehyde, and stained with mithramycin, propidium iodide, or ethidium bromide. With these DNA-fluorochromes, the fluorescence is quenched by formaldehyde less in mitotic nuclei than ... More
The potassium channel opener (-)-cromakalim prevents glutamate-induced cell death in hippocampal neurons.
AuthorsLauritzen I, De Weille JR, Lazdunski M
JournalJ Neurochem
PubMed ID9326286
'(-)-Cromakalim, a typical K+-channel opener, prevents neuronal death induced by either glucose and oxygen privation or by high (100 microM) extracellular glutamate in primary cultures of hippocampus. (-)-Cromakalim has no effect on the earliest events associated with exposure to glutamate. It does not prevent the rapid rise of intracellular Ca2+, ... More
Cold-induced apoptosis of hepatocytes: mitochondrial permeability transition triggered by nonmitochondrial chelatable iron.
AuthorsRauen U, Kerkweg U, Weisheit D, Petrat F, Sustmann R, de Groot H
JournalFree Radic Biol Med
PubMed ID14680689
'We previously described that the cold-induced apoptosis of cultured hepatocytes is mediated by an increase in the cellular chelatable iron pool. We here set out to assess whether a mitochondrial permeability transition (MPT) is involved in cold-induced apoptosis. When cultured hepatocytes were rewarmed after 18 h of cold (4 degrees ... More
Flow cytometric assessment of viability of lactic acid bacteria.
AuthorsBunthof CJ, Bloemen K, Breeuwer P, Rombouts FM, Abee T
JournalAppl Environ Microbiol
PubMed ID11319119
'The viability of lactic acid bacteria is crucial for their applications as dairy starters and as probiotics. We investigated the usefulness of flow cytometry (FCM) for viability assessment of lactic acid bacteria. The esterase substrate carboxyfluorescein diacetate (cFDA) and the dye exclusion DNA binding probes propidium iodide (PI) and TOTO-1 ... 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
Seasonal changes of semen quality and freezability in Franches-Montagnes stallions.
AuthorsJanett F, Thun R, Bettschen S, Burger D, Hassig M
JournalAnim Reprod Sci
PubMed ID12695055
'The objective of this study was to investigate seasonal changes of semen quality parameters in Franches-Montagnes stallions and to compare the freezability of ejaculates collected in autumn and winter. Experiments were performed using 15 stallions from the National Stud Farm in Avenches (Switzerland). Ejaculates were collected and evaluated every month ... More
Flow cytometric assessment of allopurinol susceptibility in Leishmania infantum promastigote.
AuthorsKamau SW, Hurtado M, Müller-Doblies UU, Grimm F, Nunez R
JournalCytometry
PubMed ID10918286
'BACKGROUND: Leishmaniasis is a major tropical and subtropical parasitic disease. Sodium stibogluconate, N-methyl -D-glucamine antimoniate, amphotericin B, pentamidine, and ketoconazole are drugs used to treat this disease. Some of these drugs cause severe adverse side effects and treatment failures are common. Allopurinol, a purine analog, has been used to treat ... More
Detection of apoptosis and DNA replication by differential labeling of DNA strand breaks with fluorochromes of different color.
AuthorsLi X, Melamed MR, Darzynkiewicz Z
JournalExp Cell Res
PubMed ID8549670
'Selective DNA strand break induction by photolysis (SBIP) at sites that contain incorporated halogenated nucleotides has been recently proposed as a means of analyzing DNA replication and detecting proliferating cells. The presence of numerous in situ DNA strand breaks is also an inherent feature of apoptotic cells. The aim of ... More
Alpha interferon accelerates lateral diffusion of Daudi cell surface differentiation antigens: measurement by fluorescence redistribution after photobleaching.
AuthorsBalint E, Aszalos A, Grimley PM
JournalBiochem Biophys Res Commun
PubMed ID2462420
'Lateral diffusion coefficients (D) of two surface differentiation antigens (sIgM and Bp35) were determined on interferon-sensitive (-IFs) or resistant (-IFr) Daudi cells by fluorescence photobleaching, using monospecific FITC-anti-IgM or PE-anti-Leu 16 probes. For untreated Daudi -IFs, mean (D) were 5.8 and 5.3 (x10(-10) cm2/sec). These increased, to 11 and 7.9 ... More