Use of SYTOX green dye in the flow cytometric analysis of bacterial phagocytosis.
AuthorsGaforio JJ, Serrano MJ, Ortega E, Algarra I, Alvarez de Cienfuegos G
JournalCytometry
PubMed ID12116370
BACKGROUND: Fluorescein isothiocyanate (FITC) is used widely to label the targets used in flow cytometric phagocytosis assays. Unfortunately, the fluorescence intensity of phagocytosed FITC-labeled targets is influenced by changes in intracellular pH level, making quantitative measurements with this fluorophore problematic. We describe the use of SYTOX green nucleic acid stain ... 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
Unique catabolic pathway of glycosphingolipids in a hydrozoan, Hydra magnipapillata, involving endoglycoceramidase.
AuthorsHoribata Y, Sakaguchi K, Okino N, Iida H, Inagaki M, Fujisawa T, Hama Y, Ito M
JournalJ Biol Chem
PubMed ID15320336
Endoglycoceramidase (EGCase; EC 3.2.1.123) is an enzyme capable of cleaving the glycosidic linkage between oligosaccharides and ceramides of various glycosphingolipids. We detected strong EGCase activity in animals belonging to Cnidaria, Mollusca, and Annelida and cloned the enzyme from a hydra, Hydra magnipapillata. The hydra EGCase, consisting of 517 amino acid ... More
Cell death during ischemia: relationship to mitochondrial depolarization and ROS generation.
Ischemia-reperfusion injury induces cell death, but the responsible mechanisms are not understood. This study examined mitochondrial depolarization and cell death during ischemia and reperfusion. Contracting cardiomyocytes were subjected to 60-min ischemia followed by 3-h reperfusion. Mitochondrial membrane potential (DeltaPsi(m)) was assessed with tetramethylrhodamine methyl ester. During ischemia, DeltaPsi(m) decreased to ... More
Up-regulation of cyclooxygenase-2 and apoptosis resistance by p38 MAPK in hypericin-mediated photodynamic therapy of human cancer cells.
AuthorsHendrickx N, Volanti C, Moens U, Seternes OM, de Witte P, Vandenheede JR, Piette J, Agostinis P
JournalJ Biol Chem
PubMed ID14557269
Photodynamic Therapy (PDT) is an approved anticancer therapy that kills cancer cells by the photochemical generation of reactive oxygen species following absorption of visible light by a photosensitizer, which selectively accumulates in tumors. We report that hypericin-mediated PDT of human cancer cells leads to up-regulation of the inducible cyclooxygenase-2 (COX-2) ... More
ago1 and dcr1, two core components of the RNA interference pathway, functionally diverge from rdp1 in regulating cell cycle events in Schizosaccharomyces pombe.
In the fission yeast Schizosaccharomyces pombe, three genes that function in the RNA interference (RNAi) pathway, ago1+, dcr1+, and rdp1+, have recently been shown to be important for timely formation of heterochromatin and accurate chromosome segregation. In the present study, we present evidence that null mutants for ago1+ and dcr1+ ... More
NKT cells are critical to initiate an inflammatory response after Pseudomonas aeruginosa ocular infection in susceptible mice.
AuthorsHazlett LD, Li Q, Liu J, McClellan S, Du W, Barrett RP
JournalJ Immunol
PubMed ID17617607
'CD4(+) T cells produce IFN-gamma contributing to corneal perforation in C57BL/6 (B6) mice after Pseudomonas aeruginosa infection. To determine the role of NK and NKT cells, infected corneas of B6 mice were dual immunolabeled. Initially, more NKT than NK cells were detected, but as disease progressed, NK cells increased, while ... More
PKC putative phosphorylation site Ser235 is required for MIP/AQP0 translocation to the plasma membrane.
AuthorsGolestaneh N, Fan J, Zelenka P, Chepelinsky AB,
JournalMol Vis
PubMed ID18523655
'PURPOSE: To investigate the functional significance of MIP/AQP0 phosphorylation at serine(235). METHODS: MIP/AQP0 expression and cellular localization was studied in rat lens epithelia explants induced to differentiate by FGF-2. MIP wild type (WT) and MIP (S235A) mutant expression plasmids were constructed and transiently expressed in RK13 cells. Subcellular localization of ... More
TGFbeta/activin/nodal signaling is necessary for the maintenance of pluripotency in human embryonic stem cells.
AuthorsJames D, Levine AJ, Besser D, Hemmati-Brivanlou A
JournalDevelopment
PubMed ID15703277
'Human embryonic stem cells (hESCs) self-renew indefinitely and give rise to derivatives of all three primary germ layers, yet little is known about the signaling cascades that govern their pluripotent character. Because it plays a prominent role in the early cell fate decisions of embryonic development, we have examined the ... More
Age-related macular degeneration. The lipofusion component N-retinyl-N-retinylidene ethanolamine detaches proapoptotic proteins from mitochondria and induces apoptosis in mammalian retinal pigment epithelial cells.
AuthorsSuter M, Remé C, Grimm C, Wenzel A, Jäättela M, Esser P, Kociok N, Leist M, Richter C
JournalJ Biol Chem
PubMed ID11006290
'10-20% of individuals over the age of 65 suffer from age-related macular degeneration (AMD), the leading cause of severe visual impairment in humans living in developed countries. The pathogenesis of this complex disease is poorly understood, and no efficient therapy or prevention exists to date. A precondition for AMD appears ... 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
Slit proteins prevent midline crossing and determine the dorsoventral position of major axonal pathways in the mammalian forebrain.
AuthorsBagri A, Marín O, Plump AS, Mak J, Pleasure SJ, Rubenstein JL, Tessier-Lavigne M
JournalNeuron
PubMed ID11804571
'We report that Slit proteins, a family of secreted chemorepellents, are crucial for the proper development of several major forebrain tracts. Mice deficient in Slit2 and, even more so, mice deficient in both Slit1 and Slit2 show significant axon guidance errors in a variety of pathways, including corticofugal, callosal, and ... More
Regulation of latent sensory hair cell precursors by glia in the zebrafish lateral line.
AuthorsGrant KA, Raible DW, Piotrowski T,
JournalNeuron
PubMed ID15629703
'The lateral line is a placodally derived mechanosensory organ in anamniotes that detects the movement of water. In zebrafish embryos, a migrating primordium deposits seven to nine clusters of sensory hair cells, or neuromasts, at intervals along the trunk. Postembryonically, neuromasts continue to be added. We show that some secondary ... More
AuthorsSnow CJ, Goody M, Kelly MW, Oster EC, Jones R, Khalil A, Henry CA,
JournalPLoS Genet
PubMed ID18833302
'Skeletal muscle morphogenesis transforms short muscle precursor cells into long, multinucleate myotubes that anchor to tendons via the myotendinous junction (MTJ). In vertebrates, a great deal is known about muscle specification as well as how somitic cells, as a cohort, generate the early myotome. However, the cellular mechanisms that generate ... More
Induction of terminal differentiation in epithelial cells requires polymerization of hensin by galectin 3.
'During terminal differentiation, epithelia become columnar and develop specialized apical membrane structures (microvilli) and functions (regulated endocytosis and exocytosis). Using a clonal intercalated epithelial cell line, we found that high seeding density induced these characteristics, whereas low density seeding maintained a protoepithelial state. When cells were plated at low density, ... More
Hepatocyte growth factor induces Wnt-independent nuclear translocation of beta-catenin after Met-beta-catenin dissociation in hepatocytes.
AuthorsMonga SP, Mars WM, Pediaditakis P, Bell A, Mulé K, Bowen WC, Wang X, Zarnegar R, Michalopoulos GK
JournalCancer Res
PubMed ID11929826
'Hepatocyte growth factor (HGF) and Wnt signaling pathways have been shown to be important in embryogenesis and carcinogenesis. The aim of this study was to elucidate the mechanism of functional similarities observed in the two pathways. We used normal rat liver, primary hepatocyte cultures and a dominant-negative Met expression system ... More
Regional effects of an antivascular endothelial growth factor receptor monoclonal antibody on receptor phosphorylation and apoptosis in human 253J B-V bladder cancer xenografts.
'Vascular endothelial growth factor (VEGF) is a key angiogenic factor in a variety of solid tumors, making it one of the most attractive therapeutic targets. VEGF promotes the proliferation, survival, and differentiation of vascular endothelial cells by stimulating autophosphorylation and activation of VEGF receptor-2 (VEGFR-2, fetal liver kinase-1, and kinase ... More
Simultaneous expression of c-Jun and p53 in retinal ganglion cells of adult rat retinal slice cultures.
AuthorsUmihira J, Lindsey JD, Weinreb RN
JournalCurr Eye Res
PubMed ID12187487
'PURPOSE: To determine whether the apoptosis-associated transcription factor c-Jun and the regulator protein p53 are expressed together during retinal ganglion cell (RGC) death in slice cultures of adult rat retina, and whether c-Jun expression or p53 expression is altered by glutamate. METHODS: Newborn rat RGCs were retrogradely labeled by Di-I ... More
Direct microscopic observation of lettuce leaf decontamination with a prototype fruit and vegetable washing solution and 1% NaCl-NaHCO3.
AuthorsTakeuchi K, Frank JF
JournalJ Food Prot
PubMed ID11510667
'Efficacy of a prototype, food-grade alkaline surfactant washing solution and 1% NaCl-NaHCO3 (pH 10.0) against Escherichia coli O157:H7 cells on lettuce leaves was evaluated. Lettuce was inoculated with 10(9) CFU/ml of E. coli O157:H7 for 24 +/- 1 h at 4 degrees C. Samples were rinsed and treated with the ... More
Lewis X-containing glycans are specific and potent competitive inhibitors of the binding of ZP3 to complementary sites on capacitated, acrosome-intact mouse sperm.
AuthorsKerr CL, Hanna WF, Shaper JH, Wright WW
JournalBiol Reprod
PubMed ID15128590
'Mammalian fertilization requires a cascade of interactions between sperm and the egg's zona pellucida (ZP). O-linked glycans on mouse glycoprotein ZP3 have been implicated in mediating one step of the fertilization process, the firm adhesion of acrosome-intact sperm to the ZP. Experiments to identify structural requirements of a sperm-binding glycan ... More
An epithelial cell destined for apoptosis signals its neighbors to extrude it by an actin- and myosin-dependent mechanism.
AuthorsRosenblatt J, Raff MC, Cramer LP
JournalCurr Biol
PubMed ID11728307
'BACKGROUND: Simple epithelia encase developing embryos and organs. Although these epithelia consist of only one or two layers of cells, they must provide tight barriers for the tissues that they envelop. Apoptosis occurring within these simple epithelia could compromise this barrier. How, then, does an epithelium remove apoptotic cells without ... More
Spreading of embryologically distinct urothelial cells is inhibited by SPARC.
AuthorsHudson AE, Feng WC, Delostrinos CF, Carmean N, Bassuk JA
JournalJ Cell Physiol
PubMed ID15389586
'The AON epitope of secreted protein acidic and rich in cysteine (SPARC) is a conserved motif expressed by human SPARC in a variety of human cell types. Through the use of a monoclonal antibody that recognizes this epitope, transitional epithelium was found to restrict expression of SPARC to the suprabasal ... More
A stress surveillance system based on calcium and nitric oxide in marine diatoms.
AuthorsVardi A, Formiggini F, Casotti R, De Martino A, Ribalet F, Miralto A, Bowler C
JournalPLoS Biol
PubMed ID16475869
'Diatoms are an important group of eukaryotic phytoplankton, responsible for about 20% of global primary productivity. Study of the functional role of chemical signaling within phytoplankton assemblages is still in its infancy although recent reports in diatoms suggest the existence of chemical-based defense strategies. Here, we demonstrate how the accurate ... More
Amiloride kills malignant glioma cells independent of its inhibition of the sodium-hydrogen exchanger.
AuthorsHegde M, Roscoe J, Cala P, Gorin F
JournalJ Pharmacol Exp Ther
PubMed ID15010500
'Previously, we demonstrated that malignant glioma cell lines have increased intracellular pH (pHi) as a result of increased activities of the type I sodium/hydrogen exchanger (NHE1). This alkalotic pHi of 7.2 to 7.4 is favorable for augmented glycolysis, DNA synthesis, and cell cycle progression. Conversely, reductions in pHi have been ... More
Anticonvulsant valproic acid inhibits cardiomyocyte differentiation of embryonic stem cells by increasing intracellular levels of reactive oxygen species.
AuthorsNa L, Wartenberg M, Nau H, Hescheler J, Sauer H
JournalBirth Defects Res A Clin Mol Teratol
PubMed ID12797459
'BACKGROUND: The anticonvulsant valproic acid (VPA) exerts teratogenic properties and has been demonstrated to cause neural tube defects and malformations of the heart. The effect of VPA on the differentiation of cardiomyocytes from pluripotent murine embryonic stem cells (ES cells) was investigated. METHODS: Embryoid bodies derived from ES cells were ... More
Formation of projection pathways from the arcuate nucleus of the hypothalamus to hypothalamic regions implicated in the neural control of feeding behavior in mice.
AuthorsBouret SG, Draper SJ, Simerly RB
JournalJ Neurosci
PubMed ID15028773
'The arcuate nucleus of the hypothalamus (ARH) is a critical component of forebrain pathways that regulate a variety of neuroendocrine functions, including an important role in relaying leptin signals to other parts of the hypothalamus. However, neonatal rodents do not lose weight in response to leptin treatment in the same ... More
Diverse microglial motility behaviors during clearance of dead cells in hippocampal slices.
AuthorsPetersen MA, Dailey ME
JournalGlia
PubMed ID15042586
'We used two-channel three-dimensional time-lapse fluorescence confocal imaging in live rat hippocampal slice cultures (1-7 days in vitro) to determine the motility behaviors of activated microglia as they engage dead and dying cells following traumatic brain tissue injury. Live microglia were labeled with a fluorescently conjugated lectin (IB(4)), and dead ... More
Flow cytometric monitoring of antibiotic-induced injury in Escherichia coli using cell-impermeant fluorescent probes.
AuthorsMortimer FC, Mason DJ, Gant VA
JournalAntimicrob Agents Chemother
PubMed ID10681337
'Three fluorescent nucleic acid binding dyes-propidium iodide, TO-PRO-1, and SYTOX green-were evaluated, and their abilities to distinguish between bacterial cells with and without an intact cytoplasmic membrane were compared. Each dye was readily able to discriminate between healthy and permeabilized cells of Escherichia coli, although SYTOX green showed a greater ... More
Abnormal development of forebrain midline glia and commissural projections in Nfia knock-out mice.
'Nuclear factor I (NFI) genes are expressed in multiple organs throughout development (Chaudhry et al., 1997; for review, see Gronostajski, 2000). All four NFI genes are expressed in embryonic mouse brain, with Nfia, Nfib, and Nfix being expressed highly in developing cortex (Chaudhry et al., 1997). Disruption of the Nfia ... More
Oral fibroblast expression of wound-inducible transcript 3.0 (wit3.0) accelerates the collagen gel contraction in vitro.
AuthorsSukotjo C, Lin A, Song K, Ogawa T, Wu B, Nishimura I
JournalJ Biol Chem
PubMed ID14527947
'Wounds of the oral mucosa show faster closure with less scar formation than skin wounds in other areas. A differentially expressed cDNA, wound-inducible transcript 3.0 (wit3.0), was isolated from oral mucosal wound in rats (Sukotjo, C., Abanmy, A. A., Ogawa, T., and Nishimura, I. (2002) J. Dent. Res. 81, 229-235). ... More
Proliferation of myogenic progenitor cells following feeding in the sub-antarctic notothenioid fish Harpagifer bispinis.
AuthorsBrodeur JC, Calvo J, Johnston IA
JournalJ Exp Biol
PubMed ID12456706
'Feeding metabolism and the activation of myogenic progenitor cells were investigated in the fast myotomal muscle of the sub-Antarctic fish Hapagifer bispinis acclimatized to either simulated summer (10 degrees C; 18 h:6 h light:dark) or simulated winter (5 degrees C; 6 h:18 h light:dark) conditions. Ingestion of a single meal ... More
The Homothorax homeoprotein activates the nuclear localization of another homeoprotein, extradenticle, and suppresses eye development in Drosophila.
AuthorsPai CY, Kuo TS, Jaw TJ, Kurant E, Chen CT, Bessarab DA, Salzberg A, Sun YH
JournalGenes Dev
PubMed ID9450936
'The Extradenticle (Exd) protein in Drosophila acts as a cofactor to homeotic proteins. Its nuclear localization is regulated. We report the cloning of the Drosophila homothorax (hth) gene, a homolog of the mouse Meis1 proto-oncogene that has a homeobox related to that of exd. Comparison with Meis1 finds two regions ... More
Multiple pathways regulate MeCP2 expression in normal brain development and exhibit defects in autism-spectrum disorders.
AuthorsSamaco RC, Nagarajan RP, Braunschweig D, LaSalle JM
JournalHum Mol Genet
PubMed ID14734626
'Rett syndrome (RTT) is a neurodevelopmental disorder caused by mutations in MECP2, encoding methyl-CpG-binding protein 2 (MeCP2). Although MECP2 is ubiquitously transcribed, MeCP2 expression is developmentally regulated and heterogeneous in neuronal subpopulations, defined as MeCP2(lo) and MeCP2(hi). To test the hypothesis that pathways affecting MeCP2 expression changes may be defective ... More
Morphogenetic domains in the yolk syncytial layer of axiating zebrafish embryos.
AuthorsD'Amico LA, Cooper MS
JournalDev Dyn
PubMed ID11748830
'The yolk syncytial layer (YSL) of the teleostean yolk cell is known to play important roles in the induction of cellular mesendoderm, as well as the patterning of dorsal tissues. To determine how this extraembryonic endodermal compartment is subdivided and morphologically transformed during early development, we have examined collective movements ... More
In vitro repression of Brca1-associated RING domain gene, Bard1, induces phenotypic changes in mammary epithelial cells.
AuthorsIrminger-Finger I, Soriano JV, Vaudan G, Montesano R, Sappino AP
JournalJ Cell Biol
PubMed ID9832560
'BRCA1-associated RING domain (BARD1) was identified as a protein interacting with the breast cancer gene product BRCA1. The identification of tumorigenic missense mutations within BRCA1 that impair the formation of BARD1-BRCA1 complexes, and of BARD1 mutations in breast carcinomas, sustain the view that BARD1 is involved in BRCA1-mediated tumor suppression. ... More
The Chlamydomonas Fus1 protein is present on the mating type plus fusion organelle and required for a critical membrane adhesion event during fusion with minus gametes.
AuthorsMisamore MJ, Gupta S, Snell WJ
JournalMol Biol Cell
PubMed ID12808049
'The molecular mechanisms of the defining event in fertilization, gamete fusion, remain poorly understood. The FUS1 gene in the unicellular, biflagellated green alga Chlamydomonas is one of the few sex-specific eukaryotic genes shown by genetic analysis to be essential for gamete fusion during fertilization. In Chlamydomonas, adhesion and fusion of ... More
Spreading dilatation in rat mesenteric arteries associated with calcium-independent endothelial cell hyperpolarization.
AuthorsTakano H, Dora KA, Spitaler MM, Garland CJ
JournalJ Physiol
PubMed ID14966304
'Both ACh and levcromakalim evoke smooth muscle cell hyperpolarization and associated relaxation in rat mesenteric resistance arteries. We investigated if they could evoke conducted vasodilatation along isolated arteries, whether this reflected spreading hyperpolarization and the possible mechanism involved. Focal micropipette application of either ACh, to stimulate endothelial cell muscarinic receptors, ... More
AuthorsSteinert S, Lee E, Tresset G, Zhang D, Hortsch R, Wetzel R, Hebbar S, Sundram JR, Kesavapany S, Boschke E, Kraut R,
JournalPLoS ONE
PubMed ID18716682
'BACKGROUND: The uptake and intracellular trafficking of sphingolipids, which self-associate into plasma membrane microdomains, is associated with many pathological conditions, including viral and toxin infection, lipid storage disease, and neurodegenerative disease. However, the means available to label the trafficking pathways of sphingolipids in live cells are extremely limited. In order ... More
Microstructural differences between single-species and dual-species biofilms of Streptococcus mutans and Veillonella parvula, before and after exposure to chlorhexidine.
AuthorsKara D, Luppens SB, van Marle J, Ozok R, ten Cate JM,
JournalFEMS Microbiol Lett
PubMed ID17403046
'Dual-species biofilms of Streptococcus mutans and Veillonella parvula are less susceptible to antimicrobials than single-species biofilms of the same microorganisms. The microstructure of single and dual-species biofilms of S. mutans and/or V. parvula was visualized to find out whether the spatial arrangement of bacteria in biofilms is related to survival ... More
Targeted inhibition of Stat3 with a decoy oligonucleotide abrogates head and neck cancer cell growth.
AuthorsLeong PL, Andrews GA, Johnson DE, Dyer KF, Xi S, Mai JC, Robbins PD, Gadiparthi S, Burke NA, Watkins SF, Grandis JR
JournalProc Natl Acad Sci U S A
PubMed ID12640143
'The transcription factor signal transducer and activator of transcription 3 (Stat3) is constitutively activated in a variety of cancers including squamous cell carcinoma of the head and neck (SCCHN). Previous investigations have demonstrated that activated Stat3 contributes to a loss of growth control and transformation. To investigate the therapeutic potential ... More
Bacterial viability and antibiotic susceptibility testing with SYTOX green nucleic acid stain.
AuthorsRoth BL, Poot M, Yue ST, Millard PJ
JournalAppl Environ Microbiol
PubMed ID9172364
'A fluorescent nucleic acid stain that does not penetrate living cells was used to assess the integrity of the plasma membranes of bacteria. SYTOX Green nucleic acid stain is an unsymmetrical cyanine dye with three positive charges that is completely excluded from live eukaryotic and prokaryotic cells. Binding of SYTOX ... More
A single unbranched S-phase DNA damage and replication fork blockage checkpoint pathway.
AuthorsMarchetti MA, Kumar S, Hartsuiker E, Maftahi M, Carr AM, Freyer GA, Burhans WC, Huberman JA
JournalProc Natl Acad Sci U S A
PubMed ID12032307
'The eukaryotic intra-S-phase checkpoint, which slows DNA synthesis in response to DNA damage, is poorly understood. Is DNA damage recognized directly, or indirectly through its effects on replication forks? Is the slowing of S phase in part because of competition between DNA synthesis and recombination/repair processes? The results of our ... More
Multiphoton excitation spectra in biological samples.
AuthorsDickinson ME, Simbuerger E, Zimmermann B, Waters CW, Fraser SE
JournalJ Biomed Opt
PubMed ID12880336
'Multiphoton microscopy is becoming a popular mode of live and fixed cell imaging. This mode of imaging offers several advantages due to the fact that fluorochrome excitation is a nonlinear event resulting in excitation only at the plane of focus. Multiphoton excitation is enhanced by the use of ultrafast lasers ... More
Networks of gold nanoparticles and bacteriophage as biological sensors and cell-targeting agents.
AuthorsSouza GR, Christianson DR, Staquicini FI, Ozawa MG, Snyder EY, Sidman RL, Miller JH, Arap W, Pasqualini R
JournalProc Natl Acad Sci U S A
PubMed ID16434473
'Biological molecular assemblies are excellent models for the development of nanoengineered systems with desirable biomedical properties. Here we report an approach for fabrication of spontaneous, biologically active molecular networks consisting of bacteriophage (phage) directly assembled with gold (Au) nanoparticles (termed Au-phage). We show that when the phage are engineered so ... More
Characteristics of microcystin production in the cell cycle of Microcystis viridis.
AuthorsKameyama K, Sugiura N, Inamori Y, Maekawa T
JournalEnviron Toxicol
PubMed ID14758590
'The correlation between the content of three microcystins (types LR, RR and YR) and the cell cycle of an axenic strain of Microcystis viridis, NIES-102, was investigated under conditions of high (16 mg L(-1)) and low (1.0 mg L(-1)) nitrate (NO(3)-N) concentrations. Each phase of the cell cycle was identified ... More
Adenovirus-facilitated nuclear translocation of adeno-associated virus type 2.
AuthorsXiao W, Warrington KH, Hearing P, Hughes J, Muzyczka N
JournalJ Virol
PubMed ID12388712
'We examined cytoplasmic trafficking and nuclear translocation of adeno-associated virus type 2 (AAV) by using Alexa Fluor 488-conjugated wild-type AAV, A20 monoclonal antibody immunocytochemistry, and subcellular fractionation techniques followed by DNA hybridization. Our results indicated that in the absence of adenovirus (Ad), AAV enters the cell rapidly and escapes from ... More
Glycogen synthase kinase 3beta-mediated apoptosis of primary cortical astrocytes involves inhibition of nuclear factor kappaB signaling.
'Recent studies have revealed a positive correlation between astrocyte apoptosis and rapid disease progression in persons with neurodegenerative diseases. Glycogen synthase kinase 3beta (GSK-3beta) is a molecular regulator of cell fate in the central nervous system and a target of the phosphatidylinositol 3-kinase (PI-3K) pathway. We have therefore examined the ... More
Characterization of protein kinase C beta isoform's action on retinoblastoma protein phosphorylation, vascular endothelial growth factor-induced endothelial cell proliferation, and retinal neovascularization.
AuthorsSuzuma K, Takahara N, Suzuma I, Isshiki K, Ueki K, Leitges M, Aiello LP, King GL
JournalProc Natl Acad Sci U S A
PubMed ID11805327
'Retinal neovascularization is a major cause of blindness and requires the activities of several signaling pathways and multiple cytokines. Activation of protein kinase C (PKC) enhances the angiogenic process and is involved in the signaling of vascular endothelial growth factor (VEGF). We have demonstrated a dramatic increase in the angiogenic ... More
Apoptotic versus necrotic characteristics of retinal ganglion cell death after partial optic nerve injury.
AuthorsBien A, Seidenbecher CI, Böckers TM, Sabel BA, Kreutz MR
JournalJ Neurotrauma
PubMed ID10098960
'We have investigated time course and characteristics of retinal ganglion cell (RGC) death after partial optic nerve injury. In situ end labeling of DNA fragments with the terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine (dUTP)-biotin nick end labeling (TUNEL) method revealed the presence of apoptotic cells on as early as 5 days ... More
Characterization of zona pellucida glycoprotein 3 (ZP3) and ZP2 binding sites on acrosome-intact mouse sperm.
AuthorsKerr CL, Hanna WF, Shaper JH, Wright WW
JournalBiol Reprod
PubMed ID12021035
'There is considerable evidence that mouse fertilization requires the binding of sperm to two of the three glycoproteins that form the zona pellucida (ZP), ZP3 and ZP2. Despite the biologic importance of this binding, no one has demonstrated that sperm express separate, saturable, and specific binding sites for ZP3 and ... More
An essential function of yeast cyclin-dependent kinase Cdc28 maintains chromosome stability.
AuthorsKitazono AA, Kron SJ
JournalJ Biol Chem
PubMed ID12359726
'Multiple surveillance pathways maintain genomic integrity in yeast during mitosis. Although the cyclin-dependent kinase Cdc28 is a well established regulator of mitotic progression, evidence for a direct role in mitotic surveillance has been lacking. We have now implicated a conserved sequence in the Cdc28 carboxyl terminus in maintaining chromosome stability ... More
Near-infrared optical imaging of epidermal growth factor receptor in breast cancer xenografts.
AuthorsKe S, Wen X, Gurfinkel M, Charnsangavej C, Wallace S, Sevick-Muraca EM, Li C
JournalCancer Res
PubMed ID14633715
'The specificity of a novel epidermal growth factor (EGF)-Cy5.5 fluorescent optical probe in the detection of EGF receptor (EGFr) was assessed using continuous-wave fluorescence imaging accomplished via an intensified charge-coupled device (CCD) camera. Human mammary MDA-MB-468 (EGFr+) and MDA-MB-435 (EGFr-) cancer cells were incubated with Cy5.5, EGF-Cy5.5, or the anti-EGFr ... More
The severe slow growth of Deltasrs2 Deltarqh1 in Schizosaccharomyces pombe is suppressed by loss of recombination and checkpoint genes.
AuthorsMaftahi M, Hope JC, Delgado-Cruzata L, Han CS, Freyer GA
JournalNucleic Acids Res
PubMed ID12409469
'Our interest in the Schizosaccharomyces pombe RecQ helicase, rqh1+, led us to investigate the function of a related putative DNA helicase, srs2+. We identified the srs2+ homolog in S.pombe, and found that srs2+ is not essential for cell viability. A Deltasrs2 Deltarqh1 double mutant grows extremely slowly with aberrant shaped ... More
Requirement for the zebrafish mid-hindbrain boundary in midbrain polarisation, mapping and confinement of the retinotectal projection.
AuthorsPicker A, Brennan C, Reifers F, Clarke JD, Holder N, Brand M
JournalDevelopment
PubMed ID10357940
'The organizer at the midbrain-hindbrain boundary (MHB organizer) has been proposed to induce and polarize the midbrain during development. We investigate the requirement for the MHB organizer in acerebellar mutants, which lack a MHB and cerebellum, but retain a tectum, and are mutant for fgf8, a candidate inducer and polarizer. ... More
Simultaneous detection of apoptosis and mitochondrial superoxide production in live cells by flow cytometry and confocal microscopy.
AuthorsMukhopadhyay P, Rajesh M, Haskó G, Hawkins BJ, Madesh M, Pacher P
JournalNat Protoc
PubMed ID17853886
'Annexin V and Sytox Green are widely used markers to evaluate apoptosis in various cell types using flow cytometry and fluorescent microscopy. Recently, a novel fluoroprobe MitoSOX Red was introduced for selective detection of superoxide in the mitochondria of live cells and was validated for confocal microscopy and flow cytometry. ... More
Fluorescence microplate-based assay for tumor necrosis factor activity using SYTOX Green stain.
AuthorsJones LJ, Singer VL
JournalAnal Biochem
PubMed ID11373072
'We have developed a simple, sensitive, fluorescence microplate-based assay for tumor necrosis factor (TNF) biological activity. The assay employs SYTOX Green nucleic acid stain to detect TNF-induced cell necrosis in actinomycin D sensitized cultured cell lines. SYTOX Green stain is a cationic unsymmetrical cyanine dye that is excluded from live ... More
Confocal imaging reveals structural detail of the cell nucleus and ascospore formation in lichenized fungi.
AuthorsWolinski H
JournalMycol Res
PubMed ID14531621
'A method for visualization of fungal nuclear structures using the DNA specific fluorochrome SYTOX Green and confocal laser scanning microscopy is presented. The staining procedure detects nuclei in different cell types with high contrast. Furthermore, the method permits generation of high-resolution confocal image stacks using optimised scanning conditions. This approach ... More
Inhibition of HSP90 in Trypanosoma cruzi induces a stress response but no stage differentiation.
AuthorsGraefe SE, Wiesgigl M, Gaworski I, Macdonald A, Clos J
JournalEukaryot Cell
PubMed ID12477794
'The 90-kDa heat shock proteins (HSP90) are important in the regulation of numerous intracellular processes in eukaryotic cells. In particular, HSP90 has been shown to be involved in the control of the cellular differentiation of the protozoan parasite Leishmania donovani. We investigated the role of HSP90 in the related parasite ... More
Prostaglandin E2 and 4-aminopyridine prevent the lipopolysaccharide-induced outwardly rectifying potassium current and interleukin-1beta production in cultured rat microglia.
AuthorsCaggiano AO, Kraig RP
JournalJ Neurochem
PubMed ID9603200
'Brain inflammation includes microglial activation and enhanced production of diffusible chemical mediators, including prostaglandin E2. Prostaglandin E2 is generally considered a proinflammatory molecule, but it also promotes neuronal survival and down-regulates some aspects of microglial activation. It remains unknown, however, if and how prostaglandin E2 prevents microglial activation. In primary ... More
Gamma-tocopherol inhibits human cancer cell cycle progression and cell proliferation by down-regulation of cyclins.
AuthorsGysin R, Azzi A, Visarius T
JournalFASEB J
PubMed ID12368234
'Effects of gamma-tocopherol on the cell cycle and proliferation were examined in human prostate carcinoma, colorectal adenocarcinoma, and osteosarcoma cells. Many epidemiological studies have suggested an anticancer activity of vitamin E, yet mechanistic studies are sparse to date. Vitamin E consists of four tocopherols (alpha-, beta-, gamma-, delta-) and the ... More
Role of superoxide, nitric oxide, and peroxynitrite in doxorubicin-induced cell death in vivo and in vitro.
AuthorsMukhopadhyay P, Rajesh M, Bátkai S, Kashiwaya Y, Haskó G, Liaudet L, Szabó C, Pacher P,
JournalAm J Physiol Heart Circ Physiol
PubMed ID19286953
'Doxorubicin (DOX) is a potent available antitumor agent; however, its clinical use is limited because of its cardiotoxicity. Cell death is a key component in DOX-induced cardiotoxicity, but its mechanisms are elusive. Here, we explore the role of superoxide, nitric oxide (NO), and peroxynitrite in DOX-induced cell death using both ... More
Identification of new leishmanicidal peptide lead structures by automated real-time monitoring of changes in intracellular ATP.
AuthorsLuque-Ortega JR, Saugar JM, Chiva C, Andreu D, Rivas L
JournalBiochem J
PubMed ID12864731
'Leishmanicidal drugs interacting stoichiometrically with parasite plasma membrane lipids, thus promoting permeability, have raised significant expectations for Leishmania chemotherapy due to their nil or very low induction of resistance. Inherent in this process is a decrease in intracellular ATP, either wasted by ionic pumps to restore membrane potential or directly ... More
Peroxisome proliferator-activated receptors and hepatic stellate cell activation.
'The present study examined the roles of peroxisome proliferator-activated receptors (PPAR) in activation of hepatic stellate cells (HSC), a pivotal event in liver fibrogenesis. RNase protection assay detected mRNA for PPARgamma1 but not that for the adipocyte-specific gamma2 isoform in HSC isolated from sham-operated rats, whereas the transcripts for neither ... More
Necrotic death pathway in Fas receptor signaling.
AuthorsMatsumura H, Shimizu Y, Ohsawa Y, Kawahara A, Uchiyama Y, Nagata S
JournalJ Cell Biol
PubMed ID11121439
'A caspase 8-deficient subline (JB6) of human Jurkat cells can be killed by the oligomerization of Fas-associated protein with death domain (FADD). This cell death process is not accompanied by caspase activation, but by necrotic morphological changes. Here, we show that the death effector domain of FADD is responsible for ... More
Enterococcus faecalis plasmid pAD1-encoded Fst toxin affects membrane permeability and alters cellular responses to lantibiotics.
AuthorsWeaver KE, Weaver DM, Wells CL, Waters CM, Gardner ME, Ehli EA
JournalJ Bacteriol
PubMed ID12644486
'Fst is a peptide toxin encoded by the par toxin-antitoxin stability determinant of Enterococcus faecalis plasmid pAD1. Intracellular overproduction of Fst resulted in simultaneous inhibition of all cellular macromolecular synthesis concomitant with cell growth inhibition and compromised the integrity of the cell membrane. Cells did not lyse or noticeably leak ... More
Agrobacterium tumefaciens integrates transfer DNA into single chromosomal sites of dimorphic fungi and yields homokaryotic progeny from multinucleate yeast.
AuthorsSullivan TD, Rooney PJ, Klein BS
JournalEukaryot Cell
PubMed ID12477790
'The dimorphic fungi Blastomyces dermatitidis and Histoplasma capsulatum cause systemic mycoses in humans and other animals. Forward genetic approaches to generating and screening mutants for biologically important phenotypes have been underutilized for these pathogens. The plant-transforming bacterium Agrobacterium tumefaciens was tested to determine whether it could transform these fungi and ... More
Effectiveness of SYTOX Green stain for bacterial viability assessment.
AuthorsLebaron P, Catala P, Parthuisot N
JournalAppl Environ Microbiol
PubMed ID9647851
'The effectiveness of SYTOX Green nucleic acid stain for measuring bacterial viability was tested on starved populations of Escherichia coli and Salmonella typhimurium. This stain underestimates the fraction of dead cells within starved populations containing cells with damaged nucleic acids or membranes. Its application to natural samples should be considered ... More
The glial sling is a migratory population of developing neurons.
AuthorsShu T, Li Y, Keller A, Richards LJ
JournalDevelopment
PubMed ID12756176
'For two decades the glial sling has been hypothesized to act as a guidance substratum for developing callosal axons. However, neither the cellular nature of the sling nor its guidance properties have ever been clearly identified. Although originally thought to be glioblasts, we show here that the subventricular zone cells ... More
The core dimerization domains of histidine kinases contain recognition specificity for the cognate response regulator.
AuthorsOhta N, Newton A
JournalJ Bacteriol
PubMed ID12867451
'Histidine kinases DivJ and PleC initiate signal transduction pathways that regulate an early cell division cycle step and the gain of motility later in the Caulobacter crescentus cell cycle, respectively. The essential single-domain response regulator DivK functions downstream of these kinases to catalyze phosphotransfer from DivJ and PleC. We have ... More
Permeabilization of fungal membranes by plant defensins inhibits fungal growth.
AuthorsThevissen K, Terras FR, Broekaert WF
JournalAppl Environ Microbiol
PubMed ID10584003
'We used an assay based on the uptake of SYTOX Green, an organic compound that fluoresces upon interaction with nucleic acids and penetrates cells with compromised plasma membranes, to investigate membrane permeabilization in fungi. Membrane permeabilization induced by plant defensins in Neurospora crassa was biphasic, depending on the plant defensin ... 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
Saccharomyces Rrm3p, a 5' to 3' DNA helicase that promotes replication fork progression through telomeric and subtelomeric DNA.
AuthorsIvessa AS, Zhou JQ, Schulz VP, Monson EK, Zakian VA
JournalGenes Dev
PubMed ID12050116
'In wild-type Saccharomyces cerevisiae, replication forks slowed during their passage through telomeric C(1-3)A/TG(1-3) tracts. This slowing was greatly exacerbated in the absence of RRM3, shown here to encode a 5'' to 3'' DNA helicase. Rrm3p-dependent fork progression was seen at a modified Chromosome VII-L telomere, at the natural X-bearing Chromosome ... More
Simultaneous monitoring of three key neuronal functions in primary neuronal cultures.
AuthorsEvans GJ, Cousin MA
JournalJ Neurosci Methods
PubMed ID17049620
'The coupling of Ca(2+) influx to synaptic vesicle (SV) recycling in nerve terminals is essential for neurotransmitter release and thus neuronal communication. Both of these parameters have been monitored using fluorescent reporter dyes such as fura-2 and FM1-43 in single central nerve terminals. However, their simultaneous monitoring has been hampered ... More
The polo box is required for multiple functions of Plx1 in mitosis.
AuthorsLiu J, Lewellyn AL, Chen LG, Maller JL
JournalJ Biol Chem
PubMed ID15016807
'Polo-like kinases comprise a family of evolutionarily conserved serine/threonine protein kinases that play multiple roles in cell cycle regulation. In addition to the N-terminal catalytic domain, polo-like kinases have one or two highly conserved C-terminal non-catalytic regions, termed polo boxes. These motifs are required for targeting these kinases to subcellular ... More
3D high-content screening for the identification of compounds that target cells in dormant tumor spheroid regions.
AuthorsWenzel C, Riefke B, Gründemann S, Krebs A, Christian S, Prinz F, Osterland M, Golfier S, Räse S, Ansari N, Esner M, Bickle M, Pampaloni F, Mattheyer C, Stelzer EH, Parczyk K, Prechtl S, Steigemann P,
Journal
PubMed ID24480576
'Cancer cells in poorly vascularized tumor regions need to adapt to an unfavorable metabolic microenvironment. As distance from supplying blood vessels increases, oxygen and nutrient concentrations decrease and cancer cells react by stopping cell cycle progression and becoming dormant. As cytostatic drugs mainly target proliferating cells, cancer cell dormancy is ... More
Cell death and proliferation in acute slices and organotypic cultures of mammalian CNS.
AuthorsLossi L, Alasia S, Salio C, Merighi A,
JournalProg Neurobiol
PubMed ID19552996
'Analysis of the interplay between cell proliferation and death has been greatly advantaged by the development of CNS slice preparations. In slices, interactions between neurons and neurons and the glial cells are fundamentally preserved in a fashion close to the in vivo situation. In parallel, these preparations offer the possibility ... More
Role of the fusion peptide and membrane-proximal domain in HIV-1 envelope glycoprotein-mediated membrane fusion.
AuthorsDimitrov AS, Rawat SS, Jiang S, Blumenthal R
JournalBiochemistry
PubMed ID14640682
'The N-terminal fusion peptide and the interfacial sequence preceding the transmembrane anchor of HIV-1 gp41 are required for viral fusion. Studies with synthetic peptides indicated that these regions function by destabilizing membranes, which is regarded as a crucial step in the membrane fusion reaction. However, it is not clear whether ... More
Recapitulation of the sexual cycle of the primary fungal pathogen Cryptococcus neoformans var. gattii: implications for an outbreak on Vancouver Island, Canada.
'Cryptococcus neoformans is a human fungal pathogen that exists as three distinct varieties or sibling species: the predominantly opportunistic pathogens C. neoformans var. neoformans (serotype D) and C. neoformans var. grubii (serotype A) and the primary pathogen C. neoformans var. gattii (serotypes B and C). While serotypes A and D ... More
Translocation of molecules into cells by pH-dependent insertion of a transmembrane helix.
AuthorsReshetnyak YK, Andreev OA, Lehnert U, Engelman DM
JournalProc Natl Acad Sci U S A
PubMed ID16608910
'We have previously observed the spontaneous, pH-dependent insertion of a water-soluble peptide to form a helix across lipid bilayers [Hunt, J. F., Rath, P., Rothschild, K. J. & Engelman, D. M. (1997) Biochemistry 36, 15177-15192]. We now use a related peptide, pH (low) insertion peptide, to translocate cargo molecules attached ... More
Immunocytochemical localization of Na+-HCO3- cotransporters and carbonic anhydrase dependence of fluid transport in corneal endothelial cells.
AuthorsDiecke FP, Wen Q, Sanchez JM, Kuang K, Fischbarg J
JournalAm J Physiol Cell Physiol
PubMed ID14960417
'In corneal endothelium, there is evidence for basolateral entry of HCO(3)(-) into corneal endothelial cells via Na(+)-HCO(3)(-) cotransporter (NBC) proteins and for net HCO(3)(-) flux from the basolateral to the apical side. However, how HCO(3)(-) exits the cells through the apical membrane is unclear. We determined that cultured corneal endothelial ... More
Identification of genes responsive to intracellular zinc depletion in the human colon adenocarcinoma cell line HT-29.
AuthorsKindermann B, Döring F, Pfaffl M, Daniel H
JournalJ Nutr
PubMed ID14704293
'Zinc is essential for the structural and functional integrity of cells and plays a pivotal role in the control of gene expression. To identify genes with altered mRNA expression level after zinc depletion, we employed oligonucleotide arrays with approximately 10,000 targets and used the human colon adenocarcinoma epithelial cell line ... More
Protein sequences and cellular factors required for polar localization of a histidine kinase in Caulobacter crescentus.
AuthorsSciochetti SA, Lane T, Ohta N, Newton A
JournalJ Bacteriol
PubMed ID12374838
'The Caulobacter crescentus sensor kinase DivJ is required for an early cell division step and localizes at the base of the newly formed stalk during the G1-to-S-phase transition when the protein is synthesized. To identify sequences within DivJ that are required for polar localization, we examined the ability of mutagenized ... More
Phagocytosis of nonapoptotic cells dying by caspase-independent mechanisms.
AuthorsHirt UA, Gantner F, Leist M
JournalJ Immunol
PubMed ID10843710
'Caspase activation, exposure of phosphatidylserine (PS) on the outer surface of the plasma membrane, and rapid phagocytic removal of dying cells are key features of apoptosis. Nonapoptotic/necrotic modes of death occur independent of caspase activation, but the role of phagocytosis is largely unknown. To address this issue, we studied phagocytosis ... More
Effect of u.v. light irradiation, starvation and heat on Escherichia coli beta-D-galactosidase activity and other potential viability parameters.
AuthorsFiksdal L, Tryland I
JournalJ Appl Microbiol
PubMed ID10432588
'The effect of u.v. light irradiation and two other types of stress (heat and starvation) on cellular functions of Escherichia coli have been studied. The severe reduction of the culturable cell number (cfu) and the direct viable count (DVC) after exposure to moderate u.v. light doses (48 mWs cm-2), was ... More
Follicle separation during Drosophila oogenesis requires the activity of the kinesin II-associated polypeptide Kap in germline cells.
AuthorsPflanz R, Peter A, Schäfer U, Jäckle H
JournalEMBO Rep
PubMed ID15088066
'Cellular localization of organelles, protein complexes and single mRNAs depends on the directed transport along microtubule tracks, a process mediated by ATP-driven molecular motor proteins of the dynein and kinesin superfamilies. Kinesin II is a heterotrimeric protein complex composed of two motor subunits and a unique nonmotor Kinesin-associated protein (Kap). ... More
Iron activates NF-kappaB in Kupffer cells.
AuthorsShe H, Xiong S, Lin M, Zandi E, Giulivi C, Tsukamoto H
JournalAm J Physiol Gastrointest Liver Physiol
PubMed ID12181188
'Iron exacerbates various types of liver injury in which nuclear factor (NF)-kappaB-driven genes are implicated. This study tested a hypothesis that iron directly elicits the signaling required for activation of NF-kappaB and stimulation of tumor necrosis factor (TNF)-alpha gene expression in Kupffer cells. Addition of Fe2+ but not Fe3+ (approximately ... More
The Xenopus LIM-homeodomain protein Xlim5 regulates the differential adhesion properties of early ectoderm cells.
AuthorsHouston DW, Wylie C
JournalDevelopment
PubMed ID12736213
'One of the earliest lineage restriction events in embryogenesis is the specification of the primary germ layers: ectoderm, mesoderm and endoderm. In Xenopus, germ layer specification occurs prior to gastrulation and requires the transcription factor VegT both for the cell-autonomous specification of endoderm and the generation of mesoderm-inducing signals. In ... More
Protoporphyrin IX photodynamic therapy for synovitis.
AuthorsKirdaite G, Lange N, Busso N, Van Den Bergh H, Kucera P, So A
JournalArthritis Rheum
PubMed ID12115245
'OBJECTIVE: To determine the conditions for synovial accumulation of protoporphyrin IX (PpIX) and photodynamic therapy (PDT)-induced synovial cytotoxicity in vitro and in vivo. METHODS: Synovial tissues were obtained from mice with antigen-induced arthritis (AIA) and incubated with different concentrations of 5-aminolevulinic acid hexyl ester (h-ALA), a PpIX precursor. Following photoexcitation, ... More
Polyphosphate loss promotes SNF/SWI- and Gcn5-dependent mitotic induction of PHO5.
AuthorsNeef DW, Kladde MP
JournalMol Cell Biol
PubMed ID12748282
'Approximately 800 transcripts in Saccharomyces cerevisiae are cell cycle regulated. The oscillation of approximately 40% of these genes, including a prominent subclass involved in nutrient acquisition, is not understood. To address this problem, we focus on the mitosis-specific activation of the phosphate-responsive promoter, PHO5. We show that the unexpected mitotic ... More
Fission yeast Clp1p phosphatase affects G2/M transition and mitotic exit through Cdc25p inactivation.
AuthorsWolfe BA, Gould KL
JournalEMBO J
PubMed ID14765109
'The Cdc14 family of phosphatases specifically reverses proline-directed phosphorylation events. In Saccharomyces cerevisiae, Cdc14p promotes Cdk1p inactivation at mitotic exit by reversing Cdk1p-dependent phosphorylations. Cdk1p is a proline-directed kinase whose activity is required in all eukaryotes for the transit into mitosis. At mitotic commitment, Cdk1p participates in its own regulation ... More
Characterization of a novel origin recognition complex-like complex: implications for DNA recognition, cell cycle control, and locus-specific gene amplification.
AuthorsMohammad M, York RD, Hommel J, Kapler GM
JournalMol Cell Biol
PubMed ID12832485
'The origin recognition complex (ORC) plays a central role in eukaryotic DNA replication. Here we describe a unique ORC-like complex in Tetrahymena thermophila, TIF4, which bound in an ATP-dependent manner to sequences required for cell cycle-controlled replication and gene amplification (ribosomal DNA [rDNA] type I elements). TIF4''s mode of DNA ... More
The Rpd3-Sin3 histone deacetylase regulates replication timing and enables intra-S origin control in Saccharomyces cerevisiae.
AuthorsAparicio JG, Viggiani CJ, Gibson DG, Aparicio OM
JournalMol Cell Biol
PubMed ID15143171
'The replication of eukaryotic genomes follows a temporally staged program, in which late origin firing often occurs within domains of altered chromatin structure(s) and silenced genes. Histone deacetylation functions in gene silencing in some late-replicating regions, prompting an investigation of the role of histone deacetylation in replication timing control in ... More
Requirement of Rrm3 helicase for repair of spontaneous DNA lesions in cells lacking Srs2 or Sgs1 helicase.
AuthorsSchmidt KH, Kolodner RD
JournalMol Cell Biol
PubMed ID15060145
'The Rrm3 DNA helicase of Saccharomyces cerevisiae interacts with proliferating cell nuclear antigen and is required for replication fork progression through ribosomal DNA repeats and subtelomeric and telomeric DNA. Here, we show that rrm3 srs2 and rrm3 sgs1 mutants, in which two different DNA helicases have been inactivated, exhibit a ... More
Early activation, motility, and homing of neonatal microglia to injured neurons does not require protein synthesis.
AuthorsKurpius D, Wilson N, Fuller L, Hoffman A, Dailey ME
JournalGlia
PubMed ID16715500
'Neuronal injury in CNS tissues induces a rapid activation and mobilization of resident microglia (MG). It is widely assumed that changes in gene expression drive the morphological transformation of MG and regulate their mobilization during activation. Here, we used acutely excised neonatal rat brain slices to test whether the morphological ... More
mRNA detection by reverse transcription-PCR for monitoring viability over time in an Enterococcus faecalis viable but nonculturable population maintained in a laboratory microcosm.
AuthorsLleò MM, Pierobon S, Tafi MC, Signoretto C, Canepari P
JournalAppl Environ Microbiol
PubMed ID11010918
'The viable but nonculturable (VBNC) state is a survival strategy adopted by bacteria when they are exposed to hostile environmental conditions. It has been shown that VBNC forms of bacteria are no longer capable of growing on conventional bacteriological media but conserve pathogenic factors and/or genes. It is thus necessary ... More
Entry is a rate-limiting step for viral infection in a Drosophila melanogaster model of pathogenesis.
AuthorsCherry S, Perrimon N
JournalNat Immunol
PubMed ID14691479
'The identification of host factors that control susceptibility to infection has been hampered by a lack of amenable genetic systems. We established an in vivo model to determine the host factors that control pathogenesis and identified viral entry as a rate-limiting step for infection. We infected Drosophila melanogaster cells and ... More
Lysine N(epsilon)-trimethylation, a tool for improving the selectivity of antimicrobial peptides.
AuthorsFernández-Reyes M, Díaz D, de la Torre BG, Cabrales-Rico A, Vallès-Miret M, Jiménez-Barbero J, Andreu D, Rivas L,
JournalJ Med Chem
PubMed ID20617807
'The effects of lysine N(epsilon)-trimethylation at selected positions of the antimicrobial cecropin A-melittin hybrid peptide KWKLFKKIGAVLKVL-amide have been studied. All five monotrimethylated, four bis-trimethylated plus the per-trimethylated analogues have been synthesized and tested for antimicrobial activity on Leishmania parasites and on Gram-positive and -negative bacteria, as well as for hemolysis ... More
Significance of mitochondrial reactive oxygen species in the generation of oxidative stress in spermatozoa.
'CONTEXT: Male infertility has been linked with the excessive generation of reactive oxygen species (ROS) by defective spermatozoa. However, the subcellular origins of this activity are unclear. OBJECTIVE: The objective of this study was to determine the importance of sperm mitochondria in creating the oxidative stress associated with defective sperm ... More
Activity-dependent neurotransmitter-receptor matching at the neuromuscular junction.
AuthorsBorodinsky LN, Spitzer NC
JournalProc Natl Acad Sci U S A
PubMed ID17190810
'Signaling in the nervous system requires matching of neurotransmitter receptors with cognate neurotransmitters at synapses. The vertebrate neuromuscular junction is the best studied cholinergic synapse, but the mechanisms by which acetylcholine is matched with acetylcholine receptors are not fully understood. Because alterations in neuronal calcium spike activity alter transmitter specification ... More
Innate gender-based proclivity in response to cytotoxicity and programmed cell death pathway.
AuthorsDu L, Bayir H, Lai Y, Zhang X, Kochanek PM, Watkins SC, Graham SH, Clark RS
JournalJ Biol Chem
PubMed ID15234982
'Many central nervous system (CNS) diseases display sexual dimorphism. Exposure to circulating sex steroids is felt to be a chief contributor to this phenomenon; however, CNS diseases of childhood and the elderly also demonstrate gender predominance and/or a sexually dimorphic response to therapies. Here we show that XY and XX ... More