Nucleosomes are exposed at the cell surface in apoptosis.
AuthorsRadic M, Marion T, Monestier M
JournalJ Immunol
PubMed ID15153485
'Apoptotic cells are considered the source of DNA, histones, and nucleoprotein complexes that drive the production of autoantibodies in systemic lupus erythematosus. However, the role of apoptotic cells in the activation of the immune system is not clear. To explore interactions that may initiate or sustain the production of anti-nuclear ... More
Peroxisome proliferator-activated receptor gamma-mediated regulation of neural stem cell proliferation and differentiation.
AuthorsWada K, Nakajima A, Katayama K, Kudo C, Shibuya A, Kubota N, Terauchi Y, Tachibana M, Miyoshi H, Kamisaki Y, Mayumi T, Kadowaki T, Blumberg RS
JournalJ Biol Chem
PubMed ID16524877
'Peroxisome proliferator-activated receptor gamma (PPARgamma) plays an important role in insulin sensitivity, tissue homeostasis, and regulating cellular functions. We found high-level expression of PPARgamma in embryo mouse brain and neural stem cells (NSCs), in contrast to extremely low levels in adult mouse brain. Here, we show that PPARgamma mediates the ... 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
Probing the kinetics of SYTOX Orange stain binding to double-stranded DNA with implications for DNA analysis.
'Rapid binding kinetics of SYTOX Orange stain with double-stranded DNA (dsDNA) was revealed on the DNA fragment sizing flow cytometer. We demonstrated for the first time that the dye molecules could be adsorbed onto the capillary surface and native DNA fragments can be dynamically stained while passing through the capillary. ... More
Droplet microfluidic technology for single-cell high-throughput screening.
AuthorsBrouzes E, Medkova M, Savenelli N, Marran D, Twardowski M, Hutchison JB, Rothberg JM, Link DR, Perrimon N, Samuels ML,
JournalProc Natl Acad Sci U S A
PubMed ID19617544
'We present a droplet-based microfluidic technology that enables high-throughput screening of single mammalian cells. This integrated platform allows for the encapsulation of single cells and reagents in independent aqueous microdroplets (1 pL to 10 nL volumes) dispersed in an immiscible carrier oil and enables the digital manipulation of these reactors ... More
Efficacy of CD40 ligand gene therapy in malignant mesothelioma.
AuthorsFriedlander PL, Delaune CL, Abadie JM, Toups M, LaCour J, Marrero L, Zhong Q, Kolls JK
JournalAm J Respir Cell Mol Biol
PubMed ID12676804
'Gene delivery of CD40 Ligand (CD40L) has shown promise in murine models of melanoma and adenocarcinoma; however, its potential for thoracic malignancies such as malignant mesothelioma remains unclear. In this study, we investigated the hypothesis that CD40L gene therapy would be effective in local and distant tumor suppression in mesothelioma ... More
(R)-alpha-lipoic acid protects retinal pigment epithelial cells from oxidative damage.
AuthorsVoloboueva LA, Liu J, Suh JH, Ames BN, Miller SS
JournalInvest Ophthalmol Vis Sci
PubMed ID16249512
'PURPOSE: To determine whether (R)-alpha-lipoic acid (LA) protects cultured human fetal retinal pigment epithelial (hfRPE) cells against oxidative injury and identify the pathways that may mediate protection. METHODS: Cultured hfRPE cells were pretreated with various concentrations of LA for 14 to 16 hours followed by treatment with a chemical oxidant, ... More
Histological analysis of GFP expression in murine bone.
AuthorsJiang X, Kalajzic Z, Maye P, Braut A, Bellizzi J, Mina M, Rowe DW
JournalJ Histochem Cytochem
PubMed ID15872052
'The power for appreciating complex cellular interactions during embryonic development using green fluorescent protein (GFP) as a visual histological marker has not been applied to adult tissues due to loss of GFP signal during paraffin embedding and a high autofluorescent background, particularly in section of bone and bone marrow. Here ... More
Deep ultraviolet mapping of intracellular protein and nucleic acid in femtograms per pixel.
'By using imaging spectrophotometry with paired images in the 200- to 280-nm wavelength range, we have directly mapped intracellular nucleic acid and protein distributions across a population of Chinese hamster ovary (CHO-K1) cells. A broadband 100× objective with a numerical aperture of 1.2 NA (glycerin immersion) and a novel laser-induced-plasma ... More
Trogocytosis by Entamoeba histolytica contributes to cell killing and tissue invasion.
AuthorsRalston KS, Solga MD, Mackey-Lawrence NM, Somlata, Bhattacharya A, Petri WA,
Journal
PubMed ID24717428
'Entamoeba histolytica is the causative agent of amoebiasis, a potentially fatal diarrhoeal disease in the developing world. The parasite was named '
Ca(2+)(cyt) negatively regulates the initiation of oocyte maturation.
AuthorsSun L, Machaca K
JournalJ Cell Biol
PubMed ID15067021
'Ca(2+) is a ubiquitous intracellular messenger that is important for cell cycle progression. Genetic and biochemical evidence support a role for Ca(2+) in mitosis. In contrast, there has been a long-standing debate as to whether Ca(2+) signals are required for oocyte meiosis. Here, we show that cytoplasmic Ca(2+) (Ca(2+)(cyt)) plays ... 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
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
Nuclear localization of pyrrole-imidazole polyamide-fluorescein conjugates in cell culture.
AuthorsBest TP, Edelson BS, Nickols NG, Dervan PB
JournalProc Natl Acad Sci U S A
PubMed ID14519850
'A series of hairpin pyrrole-imidazole polyamide-fluorescein conjugates were synthesized and assayed for cellular localization. Thirteen cell lines, representing 11 human cancers, one human transformed kidney cell line, and one murine leukemia cell line, were treated with 5 microM polyamide-fluorescein conjugates for 10-14 h, then imaged by confocal laser scanning microscopy. ... More
Real-time single-molecule observation of rolling-circle DNA replication.
We present a simple technique for visualizing replication of individual DNA molecules in real time. By attaching a rolling-circle substrate to a TIRF microscope-mounted flow chamber, we are able to monitor the progression of single-DNA synthesis events and accurately measure rates and processivities of single T7 and Escherichia coli replisomes ... More
A high-throughput, homogeneous microplate assay for agents that kill mammalian tissue culture cells.
AuthorsPierce M, Wang C, Rebentisch M, Endo M, Stump M, Kamb A
JournalJ Biomol Screen
PubMed ID12857382
Screens for cytostasis/cytoxicity have considerable value for the discovery of therapeutic agents and the investigation of the biology of apoptosis. For instance, genetic screens for proteins, protein fragments, peptides, RNAs, or chemicals that kill tissue culture cells may aid in identifying new cancer therapeutic targets. A microplate assay for cell ... More
A simple way to identify non-viable cells within living plant tissue using confocal microscopy.
AuthorsTruernit E, Haseloff J,
JournalPlant Methods
PubMed ID18573203
ABSTRACT: BACKGROUND: Plant cell death is a normal process during plant development. Mutant plants may exhibit misregulation of this process, which can lead to severe growth defects. Simple ways of visualising cell death in living plant tissues can aid the study of plant development and physiology. RESULTS: Spectral variants of ... More
High content screening of cortical neurons identifies novel regulators of axon growth.
AuthorsBlackmore MG, Moore DL, Smith RP, Goldberg JL, Bixby JL, Lemmon VP,
JournalMol Cell Neurosci
PubMed ID20159039
Neurons in the central nervous system lose their intrinsic capacity for axon regeneration as they mature, and it is widely hypothesized that changes in gene expression are responsible. Testing this hypothesis and identifying the relevant genes has been challenging because hundreds to thousands of genes are developmentally regulated in CNS ... More
Identification of a signal peptide for unconventional secretion.
AuthorsDupont E, Prochiantz A, Joliot A
JournalJ Biol Chem
PubMed ID17242404
Homeoproteins are a class of transcription factors defined by the structure of their DNA-binding domain, the homeodomain. In addition to their nuclear cell-autonomous activities, homeoproteins transfer between cells, thanks to two separate steps of secretion and internalization, which both rely on unconventional mechanisms. Internalization is driven by the third helix ... More
Disease-associated prion protein in vessel walls.
AuthorsKoperek O, Kovács GG, Ritchie D, Ironside JW, Budka H, Wick G
JournalAm J Pathol
PubMed ID12466112
Human prion diseases like Creutzfeldt-Jakob disease are infectious, inherited, or sporadic neurodegenerative disorders, characterized by the accumulation of an abnormal isoform of the host-encoded prion protein. This affects nervous tissue in sporadic Creutzfeldt-Jakob disease and, additionally, in lymphoid tissue in bovine spongiform encephalopathy-linked variant Creutzfeldt-Jakob disease. Experimental studies have established ... More
Performance assessment of DNA fragment sizing by high-sensitivity flow cytometry and pulsed-field gel electrophoresis.
AuthorsFerris MM, Yan X, Habbersett RC, Shou Y, Lemanski CL, Jett JH, Yoshida TM, Marrone BL
JournalJ Clin Microbiol
PubMed ID15131156
The sizing of restriction fragments is the chief analytical technique utilized in the production of DNA fingerprints. Few techniques have been able to compete with pulsed-field gel electrophoresis (PFGE), which is capable of discriminating among bacteria at species and strain levels by resolving restriction fragments. However, an ultrasensitive flow cytometer ... More
Development of a mechanism-based, DNA staining protocol using SYTOX orange nucleic acid stain and DNA fragment sizing flow cytometry.
Accurate measurement of single DNA fragments by DNA fragment sizing flow cytometry (FSFC) depends upon precise, stoichiometric DNA staining by the intercalating dye molecules. In this study, we determined the binding characteristics of a commercially available 532 nm wavelength-excitable dye and used this information to develop a universal DNA staining ... More
Comparison of methods for fluorescent detection of viable, dead, and total Escherichia coli O157:H7 cells in suspensions and on apples using confocal scanning laser microscopy following treatment with sanitizers.
AuthorsBurnett SL, Beuchat LR
JournalInt J Food Microbiol
PubMed ID11929169
The influence of treating Escherichia coli O157:H7 cells labeled with an enhanced green fluorescent protein (EGFP) plasmid with 20 microg/ml active chlorine, 100 mg/ml hydrogen peroxide, and 80 mg/ml acetic acid on fluorescence intensity was determined. In addition, fluorescent staining methods to differentiate viable and dead E. coli O157:H7 cells ... More
Cytolytic cells induce HMGB1 release from melanoma cell lines.
AuthorsIto N, DeMarco RA, Mailliard RB, Han J, Rabinowich H, Kalinski P, Stolz DB, Zeh HJ, Lotze MT
JournalJ Leukoc Biol
PubMed ID16968820
High mobility group box 1 (HMGB1) is one of the recently defined damage-associated molecular pattern molecules, passively released from necrotic cells and secreted by activated macrophage/monocytes. Whether cytolytic cells induce HMGB1 release from tumor cells is not known. We developed a highly sensitive method for detecting intracellular HMGB1 in tumor ... More
Compartmentalization of the broad-range phospholipase C activity to the spreading vacuole is critical for Listeria monocytogenes virulence.
AuthorsYeung PS, Na Y, Kreuder AJ, Marquis H
JournalInfect Immun
PubMed ID17060464
Listeria monocytogenes is a bacterial pathogen that multiplies in the cytosol of host cells and spreads directly from cell to cell by using an actin-based mechanism of motility. The broad-range phospholipase C (PC-PLC) of L. monocytogenes contributes to bacterial escape from vacuoles formed upon cell-to-cell spread. PC-PLC is made as ... More
Canonical transient receptor potential 1 plays a role in basic fibroblast growth factor (bFGF)/FGF receptor-1-induced Ca2+ entry and embryonic rat neural stem cell proliferation.
AuthorsFiorio Pla A, Maric D, Brazer SC, Giacobini P, Liu X, Chang YH, Ambudkar IS, Barker JL
JournalJ Neurosci
PubMed ID15758179
Basic fibroblast growth factor (bFGF) and its major receptor FGF receptor-1 (FGFR-1) play an important role in the development of the cortex. The mechanisms underlying the mitogenic role of bFGF/FGFR-1 signaling have not been elucidated. Intracellular Ca2+ concentrations ([Ca2+]i) in proliferating cortical neuroepithelial cells are markedly dependent on Ca2+ entry ... More
Interplay between mitochondria and diet mediates pathogen and stress resistance in Caenorhabditis elegans.
AuthorsRevtovich AV, Lee R, Kirienko NV
JournalPLoS Genet
PubMed ID30865620
'Diet is a crucial determinant of organismal biology; interactions between the host, its diet, and its microbiota are critical to determining the health of an organism. A variety of genetic and biochemical means were used to assay stress sensitivity in C. elegans reared on two standard laboratory diets: E. coli ... More
Proteases facilitate the endosomal escape of porcine epidemic diarrhea virus during entry into host cells.
AuthorsOh C, Kim Y, Chang KO
JournalVirus Res
PubMed ID31445102
'Exogenous and endogenous proteases play important roles in porcine epidemic diarrhea virus (PEDV) entry and replication. The roles of proteases in the viral endosomal escape and replication using trypsin (KD) or elastase (AA)-adapted US PEDV strains were studied. While PEDV KD and AA require different exogenous protease for efficient replication ... More
Ferroptosis occurs through an osmotic mechanism and propagates independently of cell rupture.
Authors
JournalNat Cell Biol
PubMed ID32868903
The use of 3D cultures of MCF-10A and MCF-12A cells by high content screening for effect-based analysis of non-genotoxic carcinogens.
AuthorsEngel A, Frenzel F, Niemann B, Braeuning A, Lampen A, Buhrke T
JournalToxicol In Vitro
PubMed ID30974152
'The human breast epithelial cell lines MCF-10A and MCF-12A form well-differentiated acinus-like structures when grown in three-dimensional matrigel culture over a period of 20?days. In the present study, both cell lines were tested for their suitability to serve as an effect-based in vitro test system for non-genotoxic carcinogens. A software ... More
Unblending of Transcriptional Condensates in Human Repeat Expansion Disease.
Authors
JournalCell
PubMed ID32386547
Skin Adipocyte Stem Cell Self-Renewal Is Regulated by a PDGFA/AKT-Signaling Axis.
Prophages encode phage-defense systems with cognate self-immunity.
Authors
JournalCell Host Microbe
PubMed ID34597593
Loss of testosterone impairs anti-tumor neutrophil function.
Authors
JournalNat Commun
PubMed ID32235862
Intercalation-Based Single-Molecule Fluorescence Assay To Study DNA Supercoil Dynamics.
Authors
JournalNano Lett
PubMed ID27356180
Distinct Roles for Condensin's Two ATPase Sites in Chromosome Condensation.
Authors
JournalMol Cell
PubMed ID31629658
Cohesin mediates DNA loop extrusion by a swing and clamp" mechanism."
Authors
JournalCell
PubMed ID34624221
Multicolor two-photon imaging of in vivo cellular pathophysiology upon influenza virus infection using the two-photon IMPRESS.
Authors
JournalNat Protoc
PubMed ID31996843
Systematic Identification of Regulators of Oxidative Stress Reveals Non-canonical Roles for Peroxisomal Import and the Pentose Phosphate Pathway.
AuthorsDubreuil MM, Morgens DW, Okumoto K, Honsho M, Contrepois K, Lee-McMullen B, Traber GM, Sood RS, Dixon SJ, Snyder MP, Fujiki Y, Bassik MC
JournalCell Rep
PubMed ID32023459
Reactive oxygen species (ROS) play critical roles in metabolism and disease, yet a comprehensive analysis of the cellular response to oxidative stress is lacking. To systematically identify regulators of oxidative stress, we conducted genome-wide Cas9/CRISPR and shRNA screens. This revealed a detailed picture of diverse pathways that control oxidative stress response, ranging ... More
Lipid-droplet-accumulating microglia represent a dysfunctional and proinflammatory state in the aging brain.
AuthorsMarschallinger J, Iram T, Zardeneta M, Lee SE, Lehallier B, Haney MS, Pluvinage JV, Mathur V, Hahn O, Morgens DW, Kim J, Tevini J, Felder TK, Wolinski H, Bertozzi CR, Bassik MC, Aigner L, Wyss-Coray T
JournalNat Neurosci
PubMed ID31959936
Microglia become progressively activated and seemingly dysfunctional with age, and genetic studies have linked these cells to the pathogenesis of a growing number of neurodegenerative diseases. Here we report a striking buildup of lipid droplets in microglia with aging in mouse and human brains. These cells, which we call 'lipid-droplet-accumulating ... More