Leishmania disease development depends on the presence of apoptotic promastigotes in the virulent inoculum.
Authorsvan Zandbergen G, Bollinger A, Wenzel A, Kamhawi S, Voll R, Klinger M, Müller A, Hölscher C, Herrmann M, Sacks D, Solbach W, Laskay T
JournalProc Natl Acad Sci U S A
PubMed ID16945916
'The obligate intracellular pathogen Leishmania major survives and multiplies in professional phagocytes. The evasion strategy to circumvent killing by host phagocytes and establish a productive infection is poorly understood. Here we report that the virulent inoculum of Leishmania promastigotes contains a high ratio of annexin A5-binding apoptotic parasites. This subpopulation ... More
Towards an understanding of apoptosis detection by SYTO dyes.
AuthorsWlodkowic D, Skommer J, Pelkonen J
JournalCytometry A
PubMed ID17200958
'BACKGROUND: SYTO probes are gaining momentum as reliable and easy to use markers of apoptotic cell death, but the phenomenon underlying reduced SYTO fluorescence in apoptotic cells as compared with normal cells is still not fully elucidated. Herein, we attempt to provide further insights into mechanisms of reduced SYTO16 fluorescence ... More
Nucleic acid dyes for detection of apoptosis in live cells.
AuthorsFrey T
JournalCytometry
PubMed ID8582249
'Apoptotic thymocytes were found to be much dimmer than normal thymocytes when stained with several nucleic acid dyes. These dyes provide a quick and simple assay for apoptosis which works for live cells and does not require a UV laser. The collection of dyes giving this staining pattern includes reagents ... More
Phosphatidylserine-dependent engulfment by macrophages of nuclei from erythroid precursor cells.
AuthorsYoshida H, Kawane K, Koike M, Mori Y, Uchiyama Y, Nagata S
JournalNature
PubMed ID16193055
'Definitive erythropoiesis usually occurs in the bone marrow or fetal liver, where erythroblasts are associated with a central macrophage in anatomical units called ''blood islands''. Late in erythropoiesis, nuclei are expelled from the erythroid precursor cells and engulfed by the macrophages in the blood island. Here we show that the ... More
Multiparameter detection of apoptosis using red-excitable SYTO probes.
'Functional assays allowing phenotypic characterization of different cell death parameters at a single-cell level are important tools for preclinical anticancer drug screening. Currently, the selection of cytometric assays is limited by the availability of fluorescent probes with overlapping spectral characteristics. Following on our earlier reports on green and orange fluorescent ... More
Differential regulation of cardiomyocyte survival and hypertrophy by MDM2, an E3 ubiquitin ligase.
AuthorsToth A, Nickson P, Qin LL, Erhardt P
JournalJ Biol Chem
PubMed ID16339144
'MDM2 is an E3 ubiquitin ligase that regulates the proteasomal degradation and activity of proteins involved in cell growth and apoptosis, including the tumor suppressors p53 and retinoblastoma and the transcription factor E2F1. Although the effect of several MDM2 targets on cardiomyocyte survival and hypertrophy has already been investigated, the ... More
Transmembrane inhibitors of P-glycoprotein, an ABC transporter.
AuthorsTarasova NI, Seth R, Tarasov SG, Kosakowska-Cholody T, Hrycyna CA, Gottesman MM, Michejda CJ
JournalJ Med Chem
PubMed ID15916428
'Drug resistance mediated by ABC transporters such as P-glycoprotein (P-gp) continues to be a major impediment to effective cancer chemotherapy. We have developed a panel of highly specific peptide inhibitors of P-gp based on the structure of the transmembrane domains of the transporter. These peptides are thought to exert their ... More
Chromosome segregation defects contribute to aneuploidy in normal neural progenitor cells.
'Recent studies based predominantly on nucleotide hybridization techniques have identified aneuploid neurons and glia in the normal brain. To substantiate these findings and address how neural aneuploidy arises, we examined individual neural progenitor cells (NPCs) undergoing mitosis. Here we report the identification of chromosomal segregation defects in normal NPCs of ... More
Neocentromere-mediated chromosome movement in maize.
AuthorsYu HG, Hiatt EN, Chan A, Sweeney M, Dawe RK
JournalJ Cell Biol
PubMed ID9362502
'Neocentromere activity is a classic example of nonkinetochore chromosome movement. In maize, neocentromeres are induced by a gene or genes on Abnormal chromosome 10 (Ab10) which causes heterochromatic knobs to move poleward at meiotic anaphase. Here we describe experiments that test how neocentromere activity affects the function of linked centromere/kinetochores ... More
Modulation of [Ca2+]i signaling dynamics and metabolism by perinuclear mitochondria in mouse parotid acinar cells.
AuthorsBruce JI, Giovannucci DR, Blinder G, Shuttleworth TJ, Yule DI
JournalJ Biol Chem
PubMed ID14699167
'Parotid acinar cells exhibit rapid cytosolic calcium signals ([Ca2+]i) that initiate in the apical region but rapidly become global in nature. These characteristic [Ca2+]i signals are important for effective fluid secretion, which critically depends on a synchronized activation of spatially separated ion fluxes. Apically restricted [Ca2+]i signals were never observed ... More
SYTO16 labelling and flow cytometry of Mycobacterium avium.
AuthorsIbrahim P, Whiteley AS, Barer MR
JournalLett Appl Microbiol
PubMed ID9449859
'Mycobacterium avium cells were harvested from agar at different stages of their growth cycle, exposed to the minimum inhibitory concentration of isoniazid (INH) for 24 h and labelled with the fluorescent nucleic acid stain SYTO16. INH exposure led to a > 10-fold increase in the intensity of labelling in the ... More
Differential contribution of cytoplasmic Ca2+ and Ca2+ influx to gamete fusion and egg activation in maize.
AuthorsAntoine AF, Faure JE, Dumas C, Feijó JA
JournalNat Cell Biol
PubMed ID11781574
'In multicellular organisms, gamete fusion triggers a set of events, collectively known as egg activation, that leads to the development of a new individual. Every species that has been studied shows at least one rise in cytoplasmic Ca2+ concentration ([Ca2+]Cyt) after gamete fusion which is believed to be involved in ... More
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
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
Resolution of mitotic cells using laser scanning cytometry.
AuthorsLuther E, Kamentsky LA
JournalCytometry
PubMed ID8900469
'A microscope-based laser scanning cytometer (LSCM) has been developed that automatically measures multiple wavelength fluorescence and light scattering of cells on a microscope slide and generates lists of cytochemical and morphological features for each of thousands of cells in a typical sample. For a sample stained with a DNA stain, ... More
Characteristics of different nucleic acid staining dyes for DNA fragment sizing by flow cytometry.
AuthorsYan X, Grace WK, Yoshida TM, Habbersett RC, Velappan N, Jett JH, Keller RA, Marrone BL
JournalAnal Chem
PubMed ID10624155
'An efficient and reliable double-stranded DNA (dsDNA) staining protocol for DNA fragment sizing by flow cytometry is presented. The protocol employs 0.8 microM of PicoGreen to label a wide range of DNA concentrations (0.5 ng/mL to 10,000 ng/mL) without regard to the solution dye/bp ratios and without initial quantification of ... More
Comparison of blue nucleic acid dyes for flow cytometric enumeration of bacteria in aquatic systems.
AuthorsLebaron P, Parthuisot N, Catala P
JournalAppl Environ Microbiol
PubMed ID9572943
'Seven blue nucleic acid dyes from Molecular Probes Inc. (SYTO-9, SYTO-11, SYTO-13, SYTO-16, SYTO-BC, SYBR-I and SYBR-II) were compared with the DAPI (4'',6-diamidino-2-phenylindole) method for flow cytometric enumeration of live and fixed bacteria in aquatic systems. It was shown that SYBR-II and SYTO-9 are the most appropriate dyes for bacterial ... More
SYTO dyes and histoproteins--myriad of applications.
AuthorsTárnok A,
JournalCytometry A
PubMed ID18496838
SYTO dyes differ from each other in one or more characteristics, including cell permeability, ?uorescence enhancement upon binding nucleic acids, excitation and emission spectra, DNA/RNA selectivity and binding af?nity. This makes them versatile tools for cell biologists. ... More
Assessment of acridine orange and SYTO 16 for in vivo imaging of the peritoneal tissues in mice.
AuthorsUdovich JA, Besselsen DG, Gmitro AF,
JournalJ Microsc
PubMed ID19397741
The effect of peritoneal injection of acridine orange and SYTO 16 in mice was investigated. Images of peritoneal tissues stained with these dyes and obtained through a confocal micro-endoscope are presented. Seventy-five Balb/c mice were split into five groups and given peritoneal injections of dye or saline. The proportions of ... More
Full developmental potential of mammalian preimplantation embryos is maintained after imaging using a spinning-disk confocal microscope.
AuthorsRoss PJ, Perez GI, Ko T, Yoo MS, Cibelli JB
JournalBiotechniques
PubMed ID17191620
Fluorescent live imaging of cells and embryos at subcellular resolution poses significant challenges for biologists due to morbidity and mortality ensuing from phototoxicity. Here we report the use of a spinning-disk confocal microscope to image mouse and bovine preimplantation embryos without impairing their developmental potential. We also present data indicating ... More
Leishmania requires Rab7-mediated degradation of endocytosed hemoglobin for their growth.
AuthorsPatel N, Singh SB, Basu SK, Mukhopadhyay A,
JournalProc Natl Acad Sci U S A
PubMed ID18319337
Leishmania is unable to synthesize heme and must acquire it from exogenous source, the mechanism of which is not known. We have shown that Leishmania endocytoses hemoglobin (Hb) and subsequently degrade it probably to generate heme. To understand how internalized Hb is degraded, we have cloned and expressed Rab7 homolog ... More
SYTO probes in the cytometry of tumor cell death.
AuthorsWlodkowic D, Skommer J, Darzynkiewicz Z,
JournalCytometry A
PubMed ID18260152
Apoptosis is a complex and finely controlled cell death process of great relevance in tissue homeostasis and pathogenesis. The majority of classical apoptotic features can be examined by flow as well as image cytometry. Therefore, cytometry has been used as a technology of choice in studies of tumor cell demise. ... More
Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development.
AuthorsChen H, Detmer SA, Ewald AJ, Griffin EE, Fraser SE, Chan DC
JournalJ Cell Biol
PubMed ID12527753
Mitochondrial morphology is determined by a dynamic equilibrium between organelle fusion and fission, but the significance of these processes in vertebrates is unknown. The mitofusins, Mfn1 and Mfn2, have been shown to affect mitochondrial morphology when overexpressed. We find that mice deficient in either Mfn1 or Mfn2 die in midgestation. ... More
Confocal imaging of microglial cell dynamics in hippocampal slice cultures.
AuthorsDailey ME, Waite M
JournalMethods
PubMed ID10356354
Methods are described for imaging the cellular dynamics of microglia in live mammalian brain slice cultures. Brain slices prepared from developing rat hippocampus are cultured for up to 2 weeks by the roller tube or static filter culture technique, stained with one or more fluorescent dyes, and imaged by scanning ... More
Flow cytometry to evaluate Theileria sergenti parasitemia using the fluorescent nucleic acid stain, SYTO16.
AuthorsYagi Y, Shiono H, Kurabayashi N, Yoshihara K, Chikayama Y
JournalCytometry
PubMed ID11042620
BACKGROUND: Although the infection of Theileria sergenti is demonstrated by intraerythrocytic localization of this parasite, much time and labor are necessary in order to determine this. We applied flow cytometry to evaluate T. sergenti parasitemia using the fluorescent nucleic acid stain method. METHODS: Peripheral blood samples from cattle infected with ... More
Quantification of T-cell-mediated apoptosis in heterogeneous leukemia populations using four-color multiparameter flow cytometry.
AuthorsWesters TM, Houtenbos I, Schuurhuis GJ, Ossenkoppele GJ, van de Loosdrecht AA
JournalCytometry A
PubMed ID15895438
BACKGROUND: The unique capacity of dendritic cells to present antigens to naive T cells is being increasingly utilized in cancer therapy. The efficacy of cell-based immunotherapy can be analyzed by determination of cytotoxic activity of T cells toward tumor cells in vitro. This study supplies a flow cytometric method to ... More
Functional cardiac cell constructs on cellulose-based scaffolding.
AuthorsEntcheva E, Bien H, Yin L, Chung CY, Farrell M, Kostov Y
JournalBiomaterials
PubMed ID15147821
Cellulose and its derivatives have been successfully employed as biomaterials in various applications, including dialysis membranes, diffusion-limiting membranes in biosensors, in vitro hollow fibers perfusion systems, surfaces for cell expansion, etc. In this study, we tested the potential of cellulose acetate (CA) and regenerated cellulose (RC) scaffolds for growing functional ... More
Safe susceptibility testing of Mycobacterium tuberculosis by flow cytometry with the fluorescent nucleic acid stain SYTO 16.
AuthorsPina-Vaz C, Costa-de-Oliveira S, Rodrigues AG
JournalJ Med Microbiol
PubMed ID15591259
The time needed to obtain susceptibility results of Mycobacterium tuberculosis using classical methodologies is still too long, and flow cytometry is a promising technique in the setting of the clinical laboratory, giving fast results. A safe, reliable and rapid method to study susceptibility to streptomycin, isoniazide, rifampicin and ethambutol is ... More
Characterization of bimodal cell death of insect cells in a rotating-wall vessel and shaker flask
In previous publications, we reported the benefits of a high-aspect rotating-wall vessel (HARV) over conventional bioreactors for insect-cell cultivation in terms of reduced medium requirements and enhanced longevity. To more fully understand the effects that HARV cultivation has on longevity, the present study characterizes the mode and kinetics of Spodoptera ... More
Highly sensitive and specific detection of P-glycoprotein function for haematological and solid tumour cells using a novel nucleic acid stain.
AuthorsBroxterman HJ, Schuurhuis GJ, Lankelma J, Oberink JW, Eekman CA, Claessen AM, Hoekman K, Poot M, Pinedo HM
JournalBr J Cancer
PubMed ID9376262
Progress in our understanding of the contribution of P-glycoprotein (P-gp)-mediated resistance to chemotherapy failure in haematological as well as solid tumours has been hampered by the lack of highly sensitive, reliable methods for the detection of P-gp function in fresh human tumour cells. The present study identifies the novel nucleic ... More
Large populations of non-clonogenic early apoptotic CD34-positive cells are present in frozen-thawed peripheral blood stem cell transplants.
AuthorsSchuurhuis GJ, Muijen MM, Oberink JW, de Boer F, Ossenkoppele GJ, Broxterman HJ
JournalBone Marrow Transplant
PubMed ID11313682
Apoptosis is the common cell death pathway which is initiated by a variety of different stimuli. The recognition of early apoptotic events would markedly improve reliability and convenience of apoptosis assays. In the present study the vital stain SytoR 16 in combination with the permeability marker 7-amino actinomycin D, (7-AAD) ... More
Discrimination of live and early apoptotic mononuclear cells by the fluorescent SYTO 16 vital dye.
AuthorsSparrow RL, Tippett E
JournalJ Immunol Methods
PubMed ID16165150
Accurate detection of apoptotic cells is important for the determination of cell viability. The aim of this study was to compare the sensitivity of the cell permeant SYTO 16 fluorescent dye for detecting early apoptotic mononuclear cells (MNCs) in normal donor blood with other apoptosis assays [i.e. Annexin-V, light scatter/7-amino-actinomycin-D ... More
Three restricted forms of Epstein-Barr virus latency counteracting apoptosis in c-myc-expressing Burkitt lymphoma cells.
AuthorsKelly GL, Milner AE, Baldwin GS, Bell AI, Rickinson AB
JournalProc Natl Acad Sci U S A
PubMed ID17001014
Epstein-Barr virus (EBV), a human herpesvirus, transforms B cell growth in vitro through expressing six virus-coded Epstein-Barr nuclear antigens (EBNAs) and two latent membrane proteins (LMPs). In many EBV-associated tumors, however, viral antigen expression is more restricted, and the aetiological role of the virus is unclear. For example, endemic Burkitt ... More
Migration of perilesional microglia after focal brain injury and modulation by CC chemokine receptor 5: an in situ time-lapse confocal imaging study.
AuthorsCarbonell WS, Murase S, Horwitz AF, Mandell JW
JournalJ Neurosci
PubMed ID16049180
Microglia rapidly become reactive in response to diverse stimuli and are thought to be prominent participants in the pathophysiology of both acute injury and chronic neurological diseases. However, mature microglial reactions to a focal lesion have not been characterized dynamically in adult vertebrate tissue. Here, we present a detailed analysis ... More
Fluorescent labeling of blood cells for evaluation of retinal and choroidal circulation.
AuthorsKhoobehi B, Peyman GA
JournalOphthalmic Surg Lasers
PubMed ID10037209
BACKGROUND AND OBJECTIVE: To develop a method of staining and tracking cells in vivo with a scanning laser ophthalmoscope (SLO) to evaluate the retinal and choroidal circulations. MATERIALS AND METHODS: Twelve pigmented male rats were used. Staining of leukocytes. Two commercially available nucleic acid stains SYTO 16 and SYTO 59 ... More
A novel murine model for assessing fetal and birth outcomes following transgestational maternal malaria infection.
AuthorsMorffy Smith CD, Russ BN, Andrew AK, Cooper CA, Moore JM
JournalSci Rep
PubMed ID31862902
'Plasmodium falciparum infection during pregnancy is a major cause of severe maternal illness and neonatal mortality. Mouse models are important for the study of gestational malaria pathogenesis. When infected with Plasmodium chabaudi chabaudi AS in early gestation, several inbred mouse strains abort at midgestation. We report here that outbred Swiss ... More
Hierarchical phosphorylation of apical membrane antigen 1 is required for efficient red blood cell invasion by malaria parasites.
AuthorsPrinz B, Harvey KL, Wilcke L, Ruch U, Engelberg K, Biller L, Lucet I, Erkelenz S, Heincke D, Spielmann T, Doerig C, Kunick C, Crabb BS, Gilson PR, Gilberger TW
JournalSci Rep
PubMed ID27698395
'Central to the pathogenesis of malaria is the proliferation of Plasmodium falciparum parasites within human erythrocytes. Parasites invade erythrocytes via a coordinated sequence of receptor-ligand interactions between the parasite and host cell. One key ligand, Apical Membrane Antigen 1 (AMA1), is a leading blood-stage vaccine and previous work indicates that ... More
Ulk1-mediated Atg5-independent macroautophagy mediates elimination of mitochondria from embryonic reticulocytes.
Authors
JournalNat Commun
PubMed ID24895007
Type IV pilus secretins have extracellular C termini.
Authors
JournalMBio
PubMed ID25805731
Scalable and Isotropic Expansion of Tissues with Simply Tunable Expansion Ratio.
Authors
JournalAdv Sci (Weinh)
PubMed ID31763149
Single-Cell RNA-Sequencing and Metabolomics Analyses Reveal the Contribution of Perivascular Adipose Tissue Stem Cells to Vascular Remodeling.
Authors
JournalArterioscler Thromb Vasc Biol
PubMed ID31340667
Microfluidic label-free bioprocessing of human reticulocytes from erythroid culture.
Authors
JournalLab Chip
PubMed ID32793940
Intelligent image-activated cell sorting 2.0.
Authors
JournalLab Chip
PubMed ID32459276
Constructing and optimizing 3D atlases from 2D data with application to the developing mouse brain.
Authors
JournalElife
PubMed ID33570495
The Na,K-ATPase-Dependent Src Kinase Signaling Changes with Mesenteric Artery Diameter.
Authors
JournalInt J Mol Sci
PubMed ID30142894
Whole-body imaging with single-cell resolution by tissue decolorization.
Authors
JournalCell
PubMed ID25417165
Versatile whole-organ/body staining and imaging based on electrolyte-gel properties of biological tissues.
Authors
JournalNat Commun
PubMed ID32341345
Mosquito C-type lectins maintain gut microbiome homeostasis.
AuthorsPang X, Xiao X, Liu Y, Zhang R, Liu J, Liu Q, Wang P, Cheng G
JournalNat Microbiol
PubMed ID27572642
The long-term evolutionary interaction between the host immune system and symbiotic bacteria determines their cooperative rather than antagonistic relationship. It is known that commensal bacteria have evolved a number of mechanisms to manipulate the mammalian host immune system and maintain homeostasis. However, the strategies employed by the microbiome to overcome ... More
DNMT3A mutations promote anthracycline resistance in acute myeloid leukemia via impaired nucleosome remodeling.
AuthorsGuryanova OA, Shank K, Spitzer B, Luciani L, Koche RP, Garrett-Bakelman FE, Ganzel C, Durham BH, Mohanty A, Hoermann G, Rivera SA, Chramiec AG, Pronier E, Bastian L, Keller MD, Tovbin D, Loizou E, Weinstein AR, Gonzalez AR, Lieu YK, Rowe JM, Pastore F, McKenney AS, Krivtsov AV, Sperr WR, Cross JR, Mason CE, Tallman MS, Arcila ME, Abdel-Wahab O, Armstrong SA, Kubicek S, Staber PB, Gönen M, Paietta EM, Melnick AM, Nimer SD, Mukherjee S, Levine RL
JournalNat Med
PubMed ID27841873
Although the majority of patients with acute myeloid leukemia (AML) initially respond to chemotherapy, many of them subsequently relapse, and the mechanistic basis for AML persistence following chemotherapy has not been determined. Recurrent somatic mutations in DNA methyltransferase 3A (DNMT3A), most frequently at arginine 882 (DNMT3A ... More
Rapid Detection of Apoptosis in Cultured Mammalian Cells.
AuthorsKudryavtsev I, Serebryakova M, Solovjeva L, Svetlova M, Firsanov D
JournalMethods Mol Biol
PubMed ID28710756
Flow cytometry is a powerful tool for the analysis of apoptosis, the process that directly determines cell fate after the action of different stresses. Here, we describe a flow cytometry method for the assessment of early and late stages of apoptosis in non-fixed cultured cells using SYTO16, DRAQ7, and PO-PRO1 ... More
Inhibition of glioblastoma dispersal by the MEK inhibitor PD0325901.
AuthorsShannon S, Jia D, Entersz I, Beelen P, Yu M, Carcione C, Carcione J, Mahtabfar A, Vaca C, Weaver M, Shreiber D, Zahn JD, Liu L, Lin H, Foty RA
JournalBMC Cancer
PubMed ID28187762
Dispersal of glioblastoma (GBM) cells leads to recurrence and poor prognosis. Accordingly, molecular pathways involved in dispersal are potential therapeutic targets. The mitogen activated protein kinase/extracellular signal regulated kinase (MAPK/ERK) pathway is commonly dysregulated in GBM, and targeting this pathway with MEK inhibitors has proven effective in controlling tumor growth. ... More
Whole-Brain Analysis of Cells and Circuits by Tissue Clearing and Light-Sheet Microscopy.
AuthorsMano T, Albanese A, Dodt HU, Erturk A, Gradinaru V, Treweek JB, Miyawaki A, Chung K, Ueda HR
JournalJ Neurosci
PubMed ID30381424
In this photo essay, we present a sampling of technologies from laboratories at the forefront of whole-brain clearing and imaging for high-resolution analysis of cell populations and neuronal circuits. The data presented here were provided for the eponymous Mini-Symposium presented at the Society for Neuroscience's 2018 annual meeting. ... More
Blockade Effects of Anti-Interferon- (IFN-)
AuthorsKrisnawati DI, Liu YC, Lee YJ, Wang YT, Chen CL, Tseng PC, Shen TJ, Lin CF