Fluorometric broad-range screening of compounds with affinity for nucleic acids.
AuthorsLiu Y,Danielsson B
JournalAnalytical chemistry
PubMed ID15828780
The C-type lectin receptor Endo180 displays internalization and recycling properties distinct from other members of the mannose receptor family.
AuthorsHoward Matthew J; Isacke Clare M;
JournalJ Biol Chem
PubMed ID12068012
Endo180/urokinase plasminogen activator receptor-associated protein together with the mannose receptor, the phospholipase A(2) receptor, and DEC-205/MR6-gp200 comprise the four members of the mannose receptor family. These receptors have a unique structural composition due to the presence of multiple C-type lectin-like domains within a single polypeptide backbone. In addition, they are ... 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
A high-throughput, cell-based screening method for siRNA and small molecule inhibitors of mTORC1 signaling using the In Cell Western technique.
AuthorsHoffman GR, Moerke NJ, Hsia M, Shamu CE, Blenis J,
JournalAssay Drug Dev Technol
PubMed ID20085456
'The mTORC1 pathway is a central regulator of cell growth, and defective mTORC1 regulation plays a causative role in a variety of human diseases, including cancer, tumor syndromes such as the tuberous sclerosis complex (TSC) and lymphangioleiomyomatosis (LAM), and metabolic diseases such as diabetes and obesity. Given the importance of ... More
Chitin is a size-dependent regulator of macrophage TNF and IL-10 production.
AuthorsDa Silva CA, Chalouni C, Williams A, Hartl D, Lee CG, Elias JA,
JournalJ Immunol
PubMed ID19265136
'Chitin is a ubiquitous polysaccharide in fungi, insects, and parasites. We hypothesized that chitin is a size-dependent regulator of innate immunity. To test this hypothesis, we characterized the effects of chitins of different sizes on murine bronchoalveolar or peritoneal macrophages. In these studies, large chitin fragments were inert, while both ... More
Conserved role of a complement-like protein in phagocytosis revealed by dsRNA knockout in cultured cells of the mosquito, Anopheles gambiae.
AuthorsLevashina EA, Moita LF, Blandin S, Vriend G, Lagueux M, Kafatos FC
JournalCell
PubMed ID11257225
'We characterize a novel hemocyte-specific acute phase glycoprotein from the malaria vector, Anopheles gambiae. It shows substantial structural and functional similarities, including the highly conserved thioester motif, to both a central component of mammalian complement system, factor C3, and to a pan-protease inhibitor, alpha2-macroglobulin. Most importantly, this protein serves as ... More
Stable reprogrammed heterokaryons form spontaneously in Purkinje neurons after bone marrow transplant.
AuthorsWeimann JM, Johansson CB, Trejo A, Blau HM
JournalNat Cell Biol
PubMed ID14562057
'Heterokaryons are the product of cell fusion without subsequent nuclear or chromosome loss. Decades of research using Sendai-virus or polyethylene glycol (PEG)-mediated fusion in tissue culture showed that the terminally differentiated state of a cell could be altered. But whether stable non-dividing heterokaryons could occur in animals has remained unclear. ... More
Binding of guanylyl cyclase activating protein 1 (GCAP1) to retinal guanylyl cyclase (RetGC1). The role of individual EF-hands.
AuthorsPeshenko IV, Olshevskaya EV, Dizhoor AM,
JournalJ Biol Chem
PubMed ID18541533
'Guanylyl cyclase activating protein 1 (GCAP1), after substitution of Ca(2+) by Mg(2+) in its EF-hands, stimulates photoreceptor guanylyl cyclase, RetGC1, in response to light. We inactivated metal binding in individual EF-hands of GCAP1 tagged with green fluorescent protein to assess their role in GCAP1 binding to RetGC1 in co-transfected HEK293 ... More
Distal renal tubular acidosis in mice that lack the forkhead transcription factor Foxi1.
AuthorsBlomqvist SR, Vidarsson H, Fitzgerald S, Johansson BR, Ollerstam A, Brown R, Persson AE, Bergström G G, Enerbäck S
JournalJ Clin Invest
PubMed ID15173882
'While macro- and microscopic kidney development appear to proceed normally in mice that lack Foxi1, electron microscopy reveals an altered ultrastructure of cells lining the distal nephron. Northern blot analyses, cRNA in situ hybridizations, and immunohistochemistry demonstrate a complete loss of expression of several anion transporters, proton pumps, and anion ... More
Different splice variants of filamin-B affect myogenesis, subcellular distribution, and determine binding to integrin [beta] subunits.
Authorsvan der Flier A, Kuikman I, Kramer D, Geerts D, Kreft M, Takafuta T, Shapiro SS, Sonnenberg A
JournalJ Cell Biol
PubMed ID11807098
'Integrins connect the extracellular matrix with the cell interior, and transduce signals through interactions of their cytoplasmic tails with cytoskeletal and signaling proteins. Using the yeast two-hybrid system, we isolated a novel splice variant (filamin-Bvar-1) of the filamentous actin cross-linking protein, filamin-B, that interacts with the cytoplasmic domain of the ... More
Visualization of the nuclear lamina in mouse anterior pituitary cells and immunocytochemical detection of lamin A/C by quick-freeze freeze-substitution electron microscopy.
AuthorsSenda T, Iizuka-Kogo A, Shimomura A
JournalJ Histochem Cytochem
PubMed ID15805424
'We examined the nuclear lamina in the quickly frozen anterior pituitary cells by electron microscopic techniques combined with freeze substitution, deep etching, and immunocytochemistry and compared it with that in the chemically fixed cells. By quick-freeze freeze-substitution electron microscopy, an electron-lucent layer, as thick as 20 nm, was revealed just ... More
Tracking and elucidating alphavirus-host protein interactions.
'Viral infections cause profound alterations in host cells. Here, we explore the interactions between proteins of the Alphavirus Sindbis and host factors during the course of mammalian cell infection. Using a mutant virus expressing the viral nsP3 protein tagged with green fluorescent protein (GFP) we directly observed nsP3 localization and ... More
Interruption of CryAB-amyloid oligomer formation by HSP22.
AuthorsSanbe A, Yamauchi J, Miyamoto Y, Fujiwara Y, Murabe M, Tanoue A
JournalJ Biol Chem
PubMed ID17092938
'An R120G missense mutation in alpha-B-crystallin (CryAB), a small heat-shock protein (HSP), causes a desmin-related cardiomyopathy (DRM) that is characterized by the formation of aggregates containing CryAB and desmin. The mutant CryAB protein leads to the formation of inclusion bodies, which contain amyloid oligomer intermediates (amyloid oligomer) in the cardiomyocytes. ... More
Hypoxia-inducible factor-1-dependent overexpression of myeloid cell factor-1 protects hypoxic cells against tert-butyl hydroperoxide-induced apoptosis.
AuthorsPiret JP, Minet E, Cosse JP, Ninane N, Debacq C, Raes M, Michiels C
JournalJ Biol Chem
PubMed ID15611089
'Increased levels of Mcl-1 (myeloid cell factor-1) have been reported in several cancers, suggesting an important role played by Mcl-1 in cancer cell survival. Mcl-1 is an anti-apoptotic protein shown to delay or block apoptosis. In this work, using semiquantitative immunofluorescence, real-time PCR, and RNase protection assay, an increase in ... More
Evidence that Sry is expressed in pre-Sertoli cells and Sertoli and granulosa cells have a common precursor.
AuthorsAlbrecht KH, Eicher EM
JournalDev Biol
PubMed ID11784049
'The expression of Sry in the undifferentiated, bipotential genital ridges of mammalian XY fetuses initiates testis development and is hypothesized to do so by directing supporting cell precursors to develop as Sertoli cells and not as granulosa cells. To directly test this hypothesis, transgenic mice expressing EGFP under the control ... More
Systematic characterization of nuclear proteome during apoptosis: a quantitative proteomic study by differential extraction and stable isotope labeling.
AuthorsHwang SI, Lundgren DH, Mayya V, Rezaul K, Cowan AE, Eng JK, Han DK
JournalMol Cell Proteomics
PubMed ID16540461
'Identification and characterization of the nuclear proteome is important for detailed understanding of multiple signaling events in eukaryotic cells. Toward this goal, we extensively characterized the nuclear proteome of human T leukemia cells by sequential extraction of nuclear proteins with different physicochemical properties using three buffer conditions. This large scale ... More
Partitioning of the matrix fraction of the Golgi apparatus during mitosis in animal cells.
AuthorsSeemann J, Pypaert M, Taguchi T, Malsam J, Warren G
JournalScience
PubMed ID11823640
'The Golgi apparatus is partitioned during mitosis in animal cells by a process of fragmentation, dispersal, and reassembly in each daughter cell. We fractionated the Golgi apparatus in vivo using the drug brefeldin A or a dominant-negative mutant of the Sar1p protein. After these treatments, Golgi enzymes moved back to ... More
Imaging and analysis of 3D tumor spheroids enriched for a cancer stem cell phenotype.
AuthorsRobertson FM, Ogasawara MA, Ye Z, Chu K, Pickei R, Debeb BG, Woodward WA, Hittelman WN, Cristofanilli M, Barsky SH,
JournalJ Biomol Screen
PubMed ID20639504
'Tumors that display a highly metastatic phenotype contain subpopulations of cells that display characteristics similar to embryonic stem cells. These cells exhibit the ability to undergo self-renewal; slowly replicate to retain a nucleoside analog label, leading to their definition as ' ... More
A new method for imaging and 3D reconstruction of mammalian cochlea by fluorescent confocal microscopy.
AuthorsHardie NA, MacDonald G, Rubel EW
JournalBrain Res
PubMed ID15053969
'Traditional methods for anatomical and morphometric studies of cochlear tissues have relied upon either microdissection of the organ of Corti or the generation of serial sections of the cochlea. Such methods are time-consuming, disruptive to three-dimensional relationships and often restrict sampling to very limited numbers of cells. We have found ... More
MTA1 interacts with MAT1, a cyclin-dependent kinase-activating kinase complex ring finger factor, and regulates estrogen receptor transactivation functions.
AuthorsTalukder AH, Mishra SK, Mandal M, Balasenthil S, Mehta S, Sahin AA, Barnes CJ, Kumar R
JournalJ Biol Chem
PubMed ID12527756
'The transcriptional activity of estrogen receptor-alpha is controlled by coregulators. MTA1 (metastasis-associated protein 1) represses estrogen receptor-alpha-driven transcription by recruiting histone deacetylases (HDACs) to the estrogen response element containing target gene chromatin in breast cancer cells. Using a yeast two-hybrid screen with the MTA1 C-terminal domain as bait, we identified ... More
PBK/TOPK, a proliferating neural progenitor-specific mitogen-activated protein kinase kinase.
AuthorsDougherty JD, Garcia AD, Nakano I, Livingstone M, Norris B, Polakiewicz R, Wexler EM, Sofroniew MV, Kornblum HI, Geschwind DH
JournalJ Neurosci
PubMed ID16291951
'We performed genomic subtraction coupled to microarray-based gene expression profiling and identified the PDZ (postsynaptic density-95/Discs large/zona occludens-1)-binding kinase/T-LAK (lymphokine-activated killer T cell) cell originating protein kinase (PBK/TOPK) as a gene highly enriched in neural stem cell cultures. Previous studies have identified PBK/TOPK as a mitogen-activated protein kinase (MAPK) kinase ... More
Ultrasensitive detection of genetically modified maize DNA by capillary gel electrophoresis with laser-induced fluorescence using different fluorescent intercalating dyes.
AuthorsGarcía-Cañas V, González R, Cifuentes A
JournalJ Agric Food Chem
PubMed ID12137467
'In this work, four different fluorescent intercalating dyes are compared for the ultrasensitive CGE-LIF detection of DNA from transgenic maize in flours. The fluorescent intercalating dyes compared are YOPRO-1, SYBR-Green-I, Ethidium bromide (EthBr), and EnhanCE. For all the four dyes optimum concentrations are established, and efficient separations of DNA fragments ... 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
Phosphatidylinositol 3-kinase-dependent extracellular calcium influx is essential for CX(3)CR1-mediated activation of the mitogen-activated protein kinase cascade.
'Fractalkine, the first member of the CX(3)C chemokine family, induces leukocyte chemotaxis through activation of its high affinity receptor, CX(3)CR1. Like other chemokine receptors, CX(3)CR1 is coupled to a pertussis toxin-sensitive heterotrimeric G(i) protein, which is necessary for rapid rise in the concentration of intracellular calcium. Using a Chinese hamster ... More
Requirement for HDM2 activity in the rapid degradation of p53 in neuroblastoma.
AuthorsIsaacs JS, Saito S, Neckers LM
JournalJ Biol Chem
PubMed ID11279110
'The wild type p53 tumor suppressor protein is rapidly degraded in normal cells by MDM2, the ubiquitin ligase that serves as the key regulator of p53 function by modulating protein stability. Cellular exposure to genotoxic stress triggers the stabilization of p53 by multiple pathways that converge upon interference with MDM2 ... More
Multiparameter flow cytometry as a tool to evaluate antipsoriatic therapy.
AuthorsGlade CP, van Erp PE, Boezeman JB, van de Kerkhof PC
JournalBr J Dermatol
PubMed ID9349331
'Objective comparison of different antipsoriatic therapies requires quantitative assessment of disease severity. However, clinical assessment with the widely used Psoriasis Area and Severity Index (PASI) introduces inaccuracy. An alternative is the quantitative analysis of different epidermal cell parameters using multiparameter flow cytometry. Our aim in the present study was to ... More
CpG oligodeoxynucleotide protection in polymicrobial sepsis is dependent on interleukin-17.
'CpG oligodeoxynucleotides (ODNs) may prevent mortality from infection. We have identified a therapeutic benefit in treating sepsis with phosphorothioate ODN sequences containing the CpG motif. Sepsis was induced in rats by cecal ligation and puncture (CLP), and treatment with CpG ODNs reduced sepsis mortality from 80% to 15% during a ... More
Visible diode lasers can be used for flow cytometric immunofluorescence and DNA analysis.
AuthorsDoornbos RM, De Grooth BG, Kraan YM, Van Der Poel CJ, Greve J
JournalCytometry
PubMed ID8187585
'This report describes a feasibility study concerning the use of a visible diode laser for two important fluorescence applications in a flow cytometer. With a 3 mW 635 nm diode laser, we performed immunofluorescence measurements using the fluorophore allophycocyanin (APC). We have measured CD8 positive lymphocytes with a two-step labeling ... 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
Involvement of inhibitory NKRs in the survival of a subset of memory-phenotype CD8+ T cells.
AuthorsUgolini S, Arpin C, Anfossi N, Walzer T, Cambiaggi A, Förster R, Lipp M, Toes RE, Melief CJ, Marvel J, Vivier E
JournalNat Immunol
PubMed ID11323697
'Inhibitory natural killer receptors (NKRs) such as killer cell immunoglobulin-like receptors (KIRs) in humans and Ly49 molecules in mice are expressed on NK cells and recognize multiple major histocompatibility (MHC) class I proteins. In humans and mice, a subset of CD8+ T cells also expresses NKRs and harbors a memory ... More
A subset of dynamic actin rearrangements in Drosophila requires the Arp2/3 complex.
AuthorsHudson AM, Cooley L
JournalJ Cell Biol
PubMed ID11854308
'The Arp2/3 complex has been shown to dramatically increase the slow spontaneous rate of actin filament nucleation in vitro, and it is known to be important for remodeling the actin cytoskeleton in vivo. We isolated and characterized loss of function mutations in genes encoding two subunits of the Drosophila Arp2/3 ... More
Ultra-thin-layer agarose gel electrophoresis II. Separation of DNA fragments on composite agarose-linear polymer matrices.
AuthorsGuttma A, Lengyel T, Szoke M, Sasvari-Szekely M
JournalJ Chromatogr A
PubMed ID10735309
'The effect of hydrophilic linear polymer additives (non-cross-linked polyacrylamide, hydroxyethyl cellulose and polyethylene oxide) on the migration behavior of double stranded DNA molecules, ranging from 200-1000 base pairs, were studied in ultra-thin-layer agarose gel electrophoresis. The detection sensitivity was found to be less than 0.1 ng/band using To-Pro-3 fluorophore labeling ... More
Expression of the integrin subunit alpha8 in murine lung development.
AuthorsWagner TE, Frevert CW, Herzog EL, Schnapp LM
JournalJ Histochem Cytochem
PubMed ID14500699
'The complex interplay between cells and extracellular matrix (ECM) proteins is critical for lung development. Integrins are key modulators of this interaction. The integrin subunit alpha 8 associates with the beta(1)-subunit to form an RGD-binding integrin. We previously showed that, in adult lung, alpha 8 is expressed in contractile interstitial ... More
Versatile low-viscosity sieving matrices for nondenaturing DNA separations using capillary array electrophoresis.
AuthorsMadabhushi RS, Vainer M, Dolnik V, Enad S, Barker DL, Harris DW, Mansfield ES
JournalElectrophoresis
PubMed ID9059830
'The high-resolution separation of double-stranded DNA (dsDNA) has important applications in physical mapping strategies and in the analysis of polymerase chain reaction (PCR) products. Although high-resolution separations of dsDNA by capillary electrophoresis (CE) have been reported, pulsed fields were required to achieve complete resolution of DNA fragments beyond 23 kilobase ... More
Pulsed electric fields for selection of hematopoietic cells and depletion of tumor cell contaminants.
'Purging of tumor cells and selection of stem cells are key technologies for enabling stem cell transplantation and stem cell gene therapy. Here we report a strategy for cell selection based on physical properties of the cells. Exposing cells to an external pulsed electric field (PEF) increases the natural potential ... More
Simultaneous flow cytometric measurement of viability and lymphocyte subset proliferation.
'Combined analysis of DNA content and immunofluorescence on single cells by flow cytometry provides information on the proliferative response of cellular sub-populations in mixed cell preparations. However, the presence of considerable numbers of dead (nonviable) cells impairs accurate flow cytometric data analysis, mainly, because dead cells can bind antibodies non-specifically ... More
Fluorescent DNA nanotags: supramolecular fluorescent labels based on intercalating dye arrays assembled on nanostructured DNA templates.
AuthorsBenvin AL, Creeger Y, Fisher GW, Ballou B, Waggoner AS, Armitage BA
JournalJ Am Chem Soc
PubMed ID17256855
'Fluorescence detection and imaging are vital technologies in the life sciences and clinical diagnostics. The key to obtaining high-resolution images and sensitive detection is to use fluorescent molecules or particles that absorb and emit visible light with high efficiency. We have synthesized supramolecular complexes consisting of a branched DNA template ... More
Cathepsin D and H2O2 stimulate degradation of thioredoxin-1: implication for endothelial cell apoptosis.
'Cathepsin D (CatD) is a lysosomal aspartic proteinase and plays an important role in the degradation of proteins and in apoptotic processes induced by oxidative stress, cytokines, and aging. All of these stimuli are potent inducers of endothelial cell apoptosis. Therefore, we investigated the role of CatD in endothelial cell ... More
Requirement of the protein B23 for nucleolar disassembly induced by the FRGY2a family proteins.
AuthorsGonda K, Wudel J, Nelson D, Katoku-Kikyo N, Reed P, Tamada H, Kikyo N
JournalJ Biol Chem
PubMed ID16415342
'In Xenopus somatic cell nuclear cloning, the nucleoli of donor nuclei rapidly and almost completely disappear in egg cytoplasm. We previously showed that the germ cell-specific proteins FRGY2a and FRGY2b were responsible for this unusually drastic nucleolar disassembly. The nucleolar disassembly occurs without inhibition of pre-rRNA transcription, a well known ... More
Clathrin is required for the function of the mitotic spindle.
AuthorsRoyle SJ, Bright NA, Lagnado L
JournalNature
PubMed ID15858577
'Clathrin has an established function in the generation of vesicles that transfer membrane and proteins around the cell. The formation of clathrin-coated vesicles occurs continuously in non-dividing cells, but is shut down during mitosis, when clathrin concentrates at the spindle apparatus. Here, we show that clathrin stabilizes fibres of the ... More
Human dopamine D4 receptor allele genotyping by ultrathin agarose gel electrophoresis with To-Pro-3 complexation.
AuthorsSzoke M, Sasvari-Szekely M, Barta C, Guttman A
JournalElectrophoresis
PubMed ID10217162
'Growing evidence shows the correlation between the allelic type of the dopamine D4 receptor and the human novelty-seeking personality trait. A sensitive, ultrathin agarose gel electrophoresis-based, high-throughput screening method was developed for genotyping the dopamine D4 receptor (D4DR) exon III 48 base pair repeat polymorphism. The efficiency of the method ... More
Notch1 expression in early lymphopoiesis influences B versus T lineage determination.
'Notch receptors regulate fate decisions in many cells. One outcome of Notch signaling is differentiation of bipotential precursors into one cell type versus another. To investigate consequences of Notch1 expression in hematolymphoid progenitors, mice were reconstituted with bone marrow (BM) transduced with retroviruses encoding a constitutively active form of Notch1. ... More
Loss of a1,6-fucosyltransferase suppressed liver regeneration: implication of core fucose in the regulation of growth factor receptor-mediated cellular signaling.
AuthorsWang Y, Fukuda T, Isaji T, Lu J, Gu W, Lee HH, Ohkubo Y, Kamada Y, Taniguchi N, Miyoshi E, Gu J,
Journal
PubMed ID25652335
'Core fucosylation is an important post-translational modification, which is catalyzed by a1,6-fucosyltransferase (Fut8). Increased expression of Fut8 has been shown in diverse carcinomas including hepatocarcinoma. In this study, we investigated the role of Fut8 expression in liver regeneration by using the 70% partial hepatectomy (PH) model, and found that Fut8 ... More
Roles of NPM2 in chromatin and nucleolar organization in oocytes and embryos.
AuthorsBurns KH, Viveiros MM, Ren Y, Wang P, DeMayo FJ, Frail DE, Eppig JJ, Matzuk MM
JournalScience
PubMed ID12714744
'Upon fertilization, remodeling of condensed maternal and paternal gamete DNA occurs to form the diploid genome. In Xenopus laevis, nucleoplasmin 2 (NPM2) decondenses sperm DNA in vitro. To study chromatin remodeling in vivo, we isolated mammalian NPM2 orthologs. Mouse NPM2 accumulates in oocyte nuclei and persists in preimplantation embryos. Npm2 ... More
Cytotoxic effects of dimethyl sulphoxide (DMSO) on cochlear organotypic cultures.
AuthorsQi W, Ding D, Salvi RJ,
JournalHear Res
PubMed ID18207679
'The amphipathic molecule dimethyl sulphoxide (DMSO) is a solvent often used to dissolve compounds applied to the inner ear; however, little is known about its potential cytotoxic side effects. To address this question, we applied 0.1-6% DMSO for 24h to cochlear organotypic cultures from postnatal day 3 rats and examined ... More
Changes in keratin 6 and keratin 10 (co-)expression in lesional and symptomless skin of spreading psoriasis.
AuthorsMommers JM, van Rossum MM, van Erp PE, van De Kerkhof PC
JournalDermatology
PubMed ID10971053
'BACKGROUND: Keratin 6 (K6) and keratin 10 (K10) are markers for epidermal hyperproliferation and differentiation, respectively, and are both expressed in the suprabasal layers of the epidermis. They may be co-expressed in different stages of the spreading psoriatic lesion, but single expression can also occur. OBJECTIVE: To investigate to what ... More
Method for identifying prostate cells in semen using flow cytometry.
AuthorsBarren RJ, Holmes EH, Boynton AL, Gregorakis A, Elgamal AA, Cobb OE, Wilson CL, Ragde H, Murphy GP
JournalProstate
PubMed ID9687990
'BACKGROUND: Prostate-specific antigen (PSA) cannot differentiate benign prostatic hyperplasia (BPH), from prostatitis, or prostate cancer in the range of 4.0-10 ng/ml. An accurate cytologic or histologic assessment is necessary to confirm the proper diagnosis. The nature of a biopsy tends to make it a selective test not frequently repeated. We ... More
The distribution of polycomb-group proteins during cell division and development in Drosophila embryos: impact on models for silencing.
AuthorsBuchenau P, Hodgson J, Strutt H, Arndt-Jovin DJ
JournalJ Cell Biol
PubMed ID9548724
'The subcellular three-dimensional distribution of three polycomb-group (PcG) proteins-polycomb, polyhomeotic and posterior sex combs-in fixed whole-mount Drosophila embryos was analyzed by multicolor confocal fluorescence microscopy. All three proteins are localized in complex patterns of 100 or more loci throughout most of the interphase nuclear volume. The rather narrow distribution of ... More
Differentiation of Phytophthora infestans sporangia from other airborne biological particles by flow cytometry.
AuthorsDay JP, Kell DB, Griffith GW
JournalAppl Environ Microbiol
PubMed ID11772606
'The ability of two different flow cytometers, the Microcyte (Optoflow) and the PAS-III (Partec), to differentiate sporangia of the late-blight pathogen Phytophthora infestans from other potential airborne particles was compared. With the PAS-III, light scatter and intrinsic fluorescence parameters could be used to differentiate sporangia from conidia of Alternaria or ... More
DNA labeling in living cells.
AuthorsMartin RM, Leonhardt H, Cardoso MC
JournalCytometry A
PubMed ID16082711
'BACKGROUND: Live cell fluorescence microscopy experiments often require visualization of the nucleus and the chromatin to determine the nuclear morphology or the localization of nuclear compartments. METHODS: We compared five different DNA dyes, TOPRO-3, TOTO-3, propidium iodide, Hoechst 33258, and DRAQ5, to test their usefulness in live cell experiments with ... More
Spectroscopic studies of the multiple binding modes of a trimethine-bridged cyanine dye with DNA.
AuthorsSovenyhazy KM, Bordelon JA, Petty JT
JournalNucleic Acids Res
PubMed ID12736305
'The interaction between DNA and a benzothiazole-quinoline cyanine dye with a trimethine bridge (TO-PRO-3) results in the formation of three noncovalent complexes. Unbound TO-PRO-3 has an absorption maximum (lambda(max)) of 632 nm, while the bound dyes (with calf thymus DNA) have electronic transitions with lambda(max) = 514 nm (complex I), ... More
A method for staining of cell nuclei in Xenopus laevis embryos with cyanine dyes for whole-mount confocal laser scanning microscopy.
Authorsde Mazière AM, Hage WJ, Ubbels GA
JournalJ Histochem Cytochem
PubMed ID8601700
'To study the cell cleavage pattern in experimentally treated Xenopus laevis blastulae, we devised a method to visualize all cell nuclei, whether in interphase or in a mitotic phase, in whole-mount embryos using confocal laser scanning microscopy. Optimal staining conditions were defined for the recently commercialized cyanine nucleic acid stain ... More
The effect of long-term treatment with tacalcitol on the psoriatic epidermis. A flow cytometric analysis.
AuthorsMommers JM, Castelijns FA, Seegers BA, Van Rossum MM, Van Hooijdonk CA, Van Erp PE, Van De Kerkhof PC
JournalBr J Dermatol
PubMed ID9767292
'During the last decade, novel analogues of 1alpha,25-dihydroxy vitamin D3 have been developed for the treatment of psoriasis. Recently, the efficacy of short-term treatment with the novel derivative tacalcitol (1alpha,24-dihydroxy vitamin D3) has been documented. However, data on the long-term effect of tacalcitol on psoriatic skin are sparse. In this ... More
The spatial distribution of inositol 1,4,5-trisphosphate receptor isoforms shapes Ca2+ waves.
AuthorsHernandez E, Leite MF, Guerra MT, Kruglov EA, Bruna-Romero O, Rodrigues MA, Gomes DA, Giordano FJ, Dranoff JA, Nathanson MH
JournalJ Biol Chem
PubMed ID17284437
'Cytosolic Ca(2+) is a versatile second messenger that can regulate multiple cellular processes simultaneously. This is accomplished in part through Ca(2+) waves and other spatial patterns of Ca(2+) signals. To investigate the mechanism responsible for the formation of Ca(2+) waves, we examined the role of inositol 1,4,5-trisphosphate receptor (InsP3R) isoforms ... More
Genetic immunization for antibody generation in research animals by intravenous delivery of plasmid DNA.
'Genetic immunization is an attractive approach to generate antibodies because native proteins are expressed in vivo with normal posttranscriptional modifications, avoiding time-consuming and costly antigen isolation or synthesis. Hydrodynamic tail or limb vein delivery of naked plasmid DNA expression vectors was used to induce antigen-specific antibodies in mice, rats, and ... More
TO-PRO-3 iodide: a novel HeNe laser-excitable DNA stain as an alternative for propidium iodide in multiparameter flow cytometry.
AuthorsVan Hooijdonk CA, Glade CP, Van Erp PE
JournalCytometry
PubMed ID7530620
'A new red emitting fluorophore, TO-PRO-3 iodide (TP3), which is best excited by an HeNe laser (633 nm), has been compared with propidium iodide (PI) for measuring relative DNA content. TP3, which has a peak absorbance at 642 nm and emission at 661 nm, has been tested on peripheral blood ... More
Pointed-end capping by tropomodulin3 negatively regulates endothelial cell motility.
AuthorsFischer RS, Fritz-Six KL, Fowler VM
JournalJ Cell Biol
PubMed ID12707310
'Actin filament pointed-end dynamics are thought to play a critical role in cell motility, yet regulation of this process remains poorly understood. We describe here a previously uncharacterized tropomodulin (Tmod) isoform, Tmod3, which is widely expressed in human tissues and is present in human microvascular endothelial cells (HMEC-1). Tmod3 is ... More
The epidermis of chronic idiopathic lichen planus during topical treatment with the vitamin D3 analogue KH 1060.
AuthorsGlade CP, Van Der Vleuten CJ, van Erp PE, De Jong EM, van de Kerkhof PC
JournalClin Exp Dermatol
PubMed ID9667102
'A parallel-group, double-blind, randomised study was performed to establish the effect of the vitamin D3 analogue KH 1060, in an ointment versus vehicle only, on the epidermal cell characteristics of chronic idiopathic lichen planus; KH 1060 also has marked immunosuppressive activity. A group of 10 patients were treated for 8 ... More
Quantification of epidermal cell populations in the centre and margin of stable psoriatic plaques.
AuthorsMommers JM, van Rossum MM, van Hooijdonk CA, van Erp PE, van de Kerkhof PC
JournalArch Dermatol Res
PubMed ID10195395
'The histological picture of psoriasis has been studied extensively. Several authors have investigated the differences between the centre and the margin of spreading plaques, because the margin is of great pathogenic interest as lesions enlarge by centrifugal expansion. However, little is known about the differences between the centre and the ... More
'We have implemented continuous-wave two-photon excitation of near-UV absorbing fluorophores in a scanning near-field optical microscope (SNOM). The 647-nm emission of an Ar-Kr mixed gas laser was used to excite the UV-absorbing DNA dyes DAPI, the bisbenzimidazole Hoechst 33342, and ethidium bromide in a shared aperture SNOM with uncoated fiber ... More
A transmembrane intracellular estrogen receptor mediates rapid cell signaling.
AuthorsRevankar CM, Cimino DF, Sklar LA, Arterburn JB, Prossnitz ER
JournalScience
PubMed ID15705806
'The steroid hormone estrogen regulates many functionally unrelated processes in numerous tissues. Although it is traditionally thought to control transcriptional activation through the classical nuclear estrogen receptors, it also initiates many rapid nongenomic signaling events. We found that of all G protein-coupled receptors characterized to date, GPR30 is uniquely localized ... More
Detection of bromodeoxyuridine incorporation by alteration of the fluorescence emission from nucleic acid binding dyes using only an argon ion laser.
AuthorsFrey T
JournalCytometry
PubMed ID7875038
'A method was developed that uses paired DNA dyes to detect the incorporation of bromodeoxyuridine (BrdUrd) into cellular DNA and requires only 488 nm excitation. The fluorescence of thiazole blue, TO-PRO-3, and LDS-751 was found to be enhanced by the presence of BrdUrd in DNA. Pairing LDS-751, thiazole blue, or ... More
A calpain unique to alveolates is essential in Plasmodium falciparum and its knockdown reveals an involvement in pre-S-phase development.
AuthorsRusso I, Oksman A, Vaupel B, Goldberg DE,
JournalProc Natl Acad Sci U S A
PubMed ID19164769
'Plasmodium falciparum encodes a single calpain that has a distinct domain composition restricted to alveolates. To evaluate the potential of this protein as a drug target, we assessed its essentiality. Both gene disruption by double cross-over and gene truncation by single cross-over recombination failed. We were also unable to achieve ... More
Editing antigen presentation: antigen transfer between human B lymphocytes and macrophages mediated by class A scavenger receptors.
AuthorsHarvey BP, Quan TE, Rudenga BJ, Roman RM, Craft J, Mamula MJ,
JournalJ Immunol
PubMed ID18768860
'B lymphocytes can function independently as efficient APCs. However, our previous studies demonstrate that both dendritic cells and macrophages are necessary to propagate immune responses initiated by B cell APCs. This finding led us to identify a process in mice whereby Ag-specific B cells transfer Ag to other APCs. In ... More
Rac1 orientates epithelial apical polarity through effects on basolateral laminin assembly.
AuthorsO'Brien LE, Jou TS, Pollack AL, Zhang Q, Hansen SH, Yurchenco P, Mostov KE
JournalNat Cell Biol
PubMed ID11533663
'Cellular polarization involves the generation of asymmetry along an intracellular axis. In a multicellular tissue, the asymmetry of individual cells must conform to the overlying architecture of the tissue. However, the mechanisms that couple cellular polarization to tissue morphogenesis are poorly understood. Here, we report that orientation of apical polarity ... More
Ectopic transgene expression in butterfly imaginal wing discs using vaccinia virus.
AuthorsLewis DL, Brunetti CR
JournalBiotechniques
PubMed ID16454039
Whole-mount immunohistochemistry on Xenopus embryos using far-red fluorescent dyes.
Cytoplasmic lipid droplets are translocated into the lumen of the Chlamydia trachomatis parasitophorous vacuole.
AuthorsCocchiaro JL, Kumar Y, Fischer ER, Hackstadt T, Valdivia RH,
JournalProc Natl Acad Sci U S A
PubMed ID18591669
The acquisition of host-derived lipids is essential for the pathogenesis of the obligate intracellular bacteria Chlamydia trachomatis. Current models of chlamydial lipid acquisition center on the fusion of Golgi-derived exocytic vesicles and endosomal multivesicular bodies with the bacteria-containing parasitophorous vacuole ( ... More
Rapid and simple quantification of bacterial cells by using a microfluidic device.
AuthorsSakamoto C, Yamaguchi N, Nasu M,
JournalAppl Environ Microbiol
PubMed ID15691978
This study investigated a microfluidic chip-based system (on-chip flow cytometry) for quantification of bacteria both in culture and in environmental samples. Bacterial numbers determined by this technique were similar to those obtained by direct microscopic count. The time required for this on-chip flow cytometry was only 30 min per 6 ... More
Photoswitching the Cytotoxic Properties of Platinum(II) Compounds.
AuthorsPresa A, Brissos RF, Caballero AB, Borilovic I, Korrodi-Gregório L, Pérez-Tomás R, Roubeau O, Gamez P,
Journal
PubMed ID25689285
The photoactivation of potential anticancer metal complexes is a hot topic of current research as it may lead to the development of more selective drugs. Photoactivated chemotherapy (PACT) with coordination compounds is usually based on a (photo)chemical reaction taking place at the metal center. Herein, a new strategy is exploited ... More
Immunohistochemical detection of soluble immunoglobulins in living mouse small intestines using an in vivo cryotechnique.
AuthorsShimo S, Saitoh S, Terada N, Ohno N, Saitoh Y, Ohno S,
JournalJ Immunol Methods
PubMed ID20691695
Some morphological changes are inevitable during immersion- or perfusion-fixation and following alcohol-dehydration for tissue preparations. Common immunostaining techniques for these specimens have some limitations to capture accurate localizations of soluble proteins in cells and tissues. In this study, to examine in situ distributions of immunoglobulins (Igs), small intestinal tissues of ... More
Chromosomal control of meiotic cell division.
AuthorsMcKim KS, Hawley RS
JournalScience
PubMed ID7502068
Chromosomes have multiple roles both in controlling the cell assembly and structure of the spindle and in determining chromosomal position on the spindle in many meiotic cells and in some types of mitotic cells. Moreover, functionally significant chromosome-microtubule interactions are not limited to the kinetochore but are also mediated by ... More
Importin beta-mediated nuclear import of fibroblast growth factor receptor: role in cell proliferation.
AuthorsReilly JF, Maher PA
JournalJ Cell Biol
PubMed ID11257130
Although growth factor receptors are generally thought to carry out their role in signal transduction at the cell surface, many of these transmembrane proteins translocate to the nucleus after ligand stimulation. Here, we show that the nuclear translocation of fibroblast growth factor receptor (FGFR)1 occurs via a mechanism distinct from ... More
Differing modes of tumour cell invasion have distinct requirements for Rho/ROCK signalling and extracellular proteolysis.
AuthorsSahai E, Marshall CJ
JournalNat Cell Biol
PubMed ID12844144
Rho family GTPases regulate the cytoskeleton and cell migration and are frequently overexpressed in tumours. Here, we identify two modes of tumour-cell motility in 3D matrices that involve different usage of Rho signalling. Rho signalling through ROCK promotes a rounded bleb-associated mode of motility that does not require pericellular proteolysis. ... More
Response to RAG-mediated VDJ cleavage by NBS1 and gamma-H2AX.
AuthorsChen HT, Bhandoola A, Difilippantonio MJ, Zhu J, Brown MJ, Tai X, Rogakou EP, Brotz TM, Bonner WM, Ried T, Nussenzweig A
JournalScience
PubMed ID11110662
Genetic disorders affecting cellular responses to DNA damage are characterized by high rates of translocations involving antigen receptor loci and increased susceptibility to lymphoid malignancies. We report that the Nijmegen breakage syndrome protein (NBS1) and histone gamma-H2AX, which associate with irradiation-induced DNA double-strand breaks (DSBs), are also found at sites ... More
Modified screen-printed electrodes for the investigation of the interaction of non-electroactive quinazoline derivatives with DNA.
AuthorsLabuda J, Bucková M, Jantová S, Stepánek I, Surugiu I, Danielsson B, Mascini M
JournalFresenius J Anal Chem
PubMed ID11225862
Five morpholino-quinazoline derivatives have been investigated voltammetrically using a competition with the tris(o-phenanthroline) cobalt(III) redox marker for the accumulation at dsDNA modified screen-printed electrodes. An association of quinazolines with DNA was observed at the modified electrodes polarized by the negative potential of -0.4 V vs. Ag/AgCl. This was confirmed by ... More
Visualizing priming of virus-specific CD8+ T cells by infected dendritic cells in vivo.
The rational design of vaccines that elicit CD8+ T cell responses requires knowledge of the identity of the antigen-presenting cell (APC), the location and time of presentation and the nature of the antigen presented by the APC. Here we address these questions for an antigen encoded by a recombinant vaccinia ... More
DNA staining for fluorescence and laser confocal microscopy.
AuthorsSuzuki T, Fujikura K, Higashiyama T, Takata K
JournalJ Histochem Cytochem
PubMed ID9010468
We examined five nucleic acid binding fluorescent dyes, propidium iodide, SYBR Green I, YO-PRO-1, TOTO-3, and TO-PRO-3, for nuclear DNA staining, visualized by fluorescence and laser confocal microscopy. The optimal concentration, co-staining of RNA, and bleaching speeds were examined. SYBR Green I and TO-PRO-3 almost preferentially stained the nuclear DNA, ... More
Microfabricated polycarbonate CE devices for DNA analysis.
AuthorsLiu Y, Ganser D, Schneider A, Liu R, Grodzinski P, Kroutchinina N
JournalAnal Chem
PubMed ID11569809
The microchip capillary electrophoresis (CE) devices were fabricated in polycarbonate (PC) plastic material by compression molding. The molded devices were enclosed utilizing thermal bonding to another PC wafer. These thermal bonds do not yield up to an applied force equivalent to 150 psi. Aqueous fluid transport inside the plastic CE ... More
Tribolium castaneum RR-1 Cuticular Protein TcCPR4 Is Required for Formation of Pore Canals in Rigid Cuticle.
AuthorsNoh MY, Muthukrishnan S, Kramer KJ, Arakane Y,
Journal
PubMed ID25664770
Insect cuticle is composed mainly of structural proteins and the polysaccharide chitin. The CPR family is the largest family of cuticle proteins (CPs), which can be further divided into three subgroups based on the presence of one of the three presumptive chitin-binding sequence motifs denoted as Rebers-Riddiford (R&R) consensus sequence ... More
AuthorsLeoni G, Neumann PA, Kamaly N, Quiros M, Nishio H, Jones HR, Sumagin R, Hilgarth RS, Alam A, Fredman G, Argyris I, Rijcken E, Kusters D, Reutelingsperger C, Perretti M, Parkos CA, Farokhzad OC, Neish AS, Nusrat A,
Journal
PubMed ID25664854
Epithelial restitution is an essential process that is required to repair barrier function at mucosal surfaces following injury. Prolonged breaches in epithelial barrier function result in inflammation and further damage; therefore, a better understanding of the epithelial restitution process has potential for improving the development of therapeutics. In this work, ... More
NK cell killer Ig-like receptor repertoire acquisition and maturation are strongly modulated by HLA class I molecules.
The interaction between clonally distributed inhibitory receptors and their activating counterparts on NK cells and HLA class I molecules defines NK cell functions, but the role of HLA class I ligands in the acquisition of their receptors during NK development is still unclear. Although some studies demonstrated that HLA-C affects ... More
Differential localization of the vacuolar H+ pump with G subunit isoforms (G1 and G2) in mouse neurons.
AuthorsMurata Y, Sun-Wada GH, Yoshimizu T, Yamamoto A, Wada Y, Futai M,
JournalJ Biol Chem
PubMed ID12133826
Vacuolar H(+)-ATPases (V-ATPases), a family of multimeric proton pumps, are involved in a wide variety of physiological processes. We have identified two mouse genes, Atp6g1 and Atp6g2, encoding the G1 and G2 isoforms of the V-ATPase G subunit, respectively. G1 was distributed ubiquitously in the tissues examined, whereas G2 was ... More
Targeting lentiviral vector expression to hepatocytes limits transgene-specific immune response and establishes long-term expression of human antihemophilic factor IX in mice.
AuthorsFollenzi A, Battaglia M, Lombardo A, Annoni A, Roncarolo MG, Naldini L,
JournalBlood
PubMed ID14701690
Stable gene replacement by in vivo administration of lentiviral vectors (LVs) has therapeutic potential for metabolic disorders and other systemic diseases. We studied the expression of intracellular and secreted proteins by LVs in immunocompetent mice. Liver, spleen, and bone marrow cells were efficiently transduced. However, transgene expression, driven by a ... More
Protocols to detect senescence-associated beta-galactosidase (SA-betagal) activity, a biomarker of senescent cells in culture and in vivo.
AuthorsDebacq-Chainiaux F, Erusalimsky JD, Campisi J, Toussaint O,
JournalNat Protoc
PubMed ID20010931
Normal cells can permanently lose the ability to proliferate when challenged by potentially oncogenic stress, a process termed cellular senescence. Senescence-associated beta-galactosidase (SA-betagal) activity, detectable at pH 6.0, permits the identification of senescent cells in culture and mammalian tissues. Here we describe first a cytochemical protocol suitable for the histochemical ... More
Fluorescent stains for quantification of DNA by confocal laser scanning microscopy in 3-D.
AuthorsPloeger LS, Dullens HF, Huisman A, van Diest PJ,
JournalBiotech Histochem
PubMed ID18568680
Confocal microscopy requires the use of fluorophores to visualize structures of interest within a specimen. To perform reliable measurements of the intensity of fluorescence, the stain should be specific, penetrate well into tissue sections, and bind stoichiometrically. Furthermore, emission must be linear with respect to DNA content and brightness, and ... More
An endothelial cell genetic screen identifies the GTPase Rem2 as a suppressor of p19ARF expression that promotes endothelial cell proliferation and angiogenesis.
AuthorsBierings R, Beato M, Edel MJ,
JournalJ Biol Chem
PubMed ID18056257
Angiogenesis requires an increase in endothelial cell proliferation to support an increase in mass of blood vessels. We designed an in vitro endothelial cell model to functionally screen for genes that regulate endothelial cell proliferation. A gain of function screen for genes that bypass p53 endothelial cell arrest identified Rem2, ... More
Fluorescence assay for mitochondrial permeability transition in cardiomyocytes cultured in a microtiter plate.
AuthorsChristensen ML, Braunstein TH, Treiman M,
JournalAnal Biochem
PubMed ID18417075
Mitochondrial permeability transition pore (MPTP) is a voltage-dependent, large-conductance channel of the inner mitochondrial membrane with an important role in a range of pathophysiological conditions. To facilitate studies of pharmacological pore modulation, we describe an assay in a model using neonatal cardiomyocytes in a 96-well microtiter plate format. In the ... More
Simultaneous analysis of reactive oxygen species and reduced glutathione content in living cells by polychromatic flow cytometry.
AuthorsCossarizza A, Ferraresi R, Troiano L, Roat E, Gibellini L, Bertoncelli L, Nasi M, Pinti M,
JournalNat Protoc
PubMed ID20010930
Reactive oxygen species (ROS) are continuously produced in the cell as a consequence of aerobic metabolism, and are controlled by several antioxidant mechanisms. An accurate measurement of ROS is essential to evaluate the redox status of the cell, or the effects of molecules with the pro-oxidant or antioxidant activity. Here ... More
S-glutathionylation impairs signal transducer and activator of transcription 3 activation and signaling.
AuthorsXie Y, Kole S, Precht P, Pazin MJ, Bernier M,
JournalEndocrinology
PubMed ID18988672
S-glutathionylation is a physiological, reversible protein modification of cysteine residues with glutathione in response to mild oxidative stress. Because the key cell growth regulator signal transducer and activator of transcription (STAT) 3 is particularly susceptible to redox regulation, we hypothesized that oxidative modification of cysteine residues of STAT3 by S-glutathionylation ... More
Distinct populations of quiescent and proliferative pancreatic beta-cells identified by HOTcre mediated labeling.
AuthorsHesselson D, Anderson RM, Beinat M, Stainier DY,
JournalProc Natl Acad Sci U S A
PubMed ID19706417
Pancreatic beta-cells are critical regulators of glucose homeostasis, and they vary dramatically in their glucose stimulated metabolic response and levels of insulin secretion. It is unclear whether these parameters are influenced by the developmental origin of individual beta-cells. Using HOTcre, a Cre-based genetic switch that uses heat-induction to precisely control ... More
Dkk-1 inhibits intestinal epithelial cell migration by attenuating directional polarization of leading edge cells.
AuthorsKoch S, Capaldo CT, Samarin S, Nava P, Neumaier I, Skerra A, Sacks DB, Parkos CA, Nusrat A,
JournalMol Biol Cell
PubMed ID19776352
Wnt signaling pathways regulate proliferation, motility, and survival in a variety of human cell types. Dickkopf-1 (Dkk-1) is a secreted Wnt antagonist that has been proposed to regulate tissue homeostasis in the intestine. In this report, we show that Dkk-1 is secreted by intestinal epithelial cells after wounding and that ... More
Chick embryo proliferation studies using EdU labeling.
AuthorsWarren M, Puskarczyk K, Chapman SC,
JournalDev Dyn
PubMed ID19253396
Cell proliferation studies are an important experimental tool. The most commonly used thymidine analogues, tritiated thymidine and bromodeoxyuridine (BrdU) label cells during S-phase. Both methods have significant drawbacks: low sensitivity in the case of tritiated thymidine and a denaturation step during BrdU detection that destroys most cellular epitopes, requiring careful ... More
Rac-mediated macropinocytosis is a critical route for naked plasmid DNA transfer in mice.
AuthorsFumoto S, Nishi J, Ishii H, Wang X, Miyamoto H, Yoshikawa N, Nakashima M, Nakamura J, Nishida K,
JournalMol Pharm
PubMed ID19492848
We have recently discovered the potential for in vivo naked plasmid DNA (pDNA) transfer into gastric serosal surface cells in mice. As pDNA are huge molecules, the mechanism of gene transfer without carriers and physical forces is of great biological interest. The endocytic route for naked pDNA transfer into gastric ... More
GPR55 is a cannabinoid receptor that increases intracellular calcium and inhibits M current.
AuthorsLauckner JE, Jensen JB, Chen HY, Lu HC, Hille B, Mackie K,
JournalProc Natl Acad Sci U S A
PubMed ID18263732
The CB(1) cannabinoid receptor mediates many of the psychoactive effects of Delta(9)THC, the principal active component of cannabis. However, ample evidence suggests that additional non-CB(1)/CB(2) receptors may contribute to the behavioral, vascular, and immunological actions of Delta(9)THC and endogenous cannabinoids. Here, we provide further evidence that GPR55, a G protein-coupled ... More
The transmembrane domain of the severe acute respiratory syndrome coronavirus ORF7b protein is necessary and sufficient for its retention in the Golgi complex.
AuthorsSchaecher SR, Diamond MS, Pekosz A,
JournalJ Virol
PubMed ID18632859
The severe acute respiratory syndrome coronavirus (SARS-CoV) ORF7b (also called 7b) protein is an integral membrane protein that is translated from a bicistronic open reading frame encoded within subgenomic RNA 7. When expressed independently or during virus infection, ORF7b accumulates in the Golgi compartment, colocalizing with both cis- and trans-Golgi ... More
Inducible gene targeting in the neonatal vasculature and analysis of retinal angiogenesis in mice.
AuthorsPitulescu ME, Schmidt I, Benedito R, Adams RH,
JournalNat Protoc
PubMed ID20725067
The retina is a powerful experimental system for the analysis of angiogenic blood vessel growth in the postnatal organisms. The three-dimensional architecture of the vessel network and processes as diverse as endothelial cell (EC) proliferation, sprouting, perivascular cell recruitment, vessel remodeling or maturation can be investigated at high resolution. The ... More
Insoluble, speckled cytosolic distribution of retinoic acid receptor alpha protein as a marker of hepatic stellate cell activation in vitro.
AuthorsMezaki Y, Yamaguchi N, Yoshikawa K, Miura M, Imai K, Itoh H, Senoo H,
JournalJ Histochem Cytochem
PubMed ID19332432
Hepatic stellate cells (HSCs) are the major site of retinoid storage, and their activation is a key process in liver fibrogenesis. We have previously shown that expression of the retinoic acid receptor alpha (RARalpha) is upregulated in activated rat HSCs at a posttranscriptional level and that these RARalpha proteins showed ... More
Vertical silicon nanowires as a universal platform for delivering biomolecules into living cells.
AuthorsShalek AK, Robinson JT, Karp ES, Lee JS, Ahn DR, Yoon MH, Sutton A, Jorgolli M, Gertner RS, Gujral TS, Macbeath G, Yang EG, Park H,
JournalProc Natl Acad Sci U S A
PubMed ID20080678
A generalized platform for introducing a diverse range of biomolecules into living cells in high-throughput could transform how complex cellular processes are probed and analyzed. Here, we demonstrate spatially localized, efficient, and universal delivery of biomolecules into immortalized and primary mammalian cells using surface-modified vertical silicon nanowires. The method relies ... More
Targeted delivery of proteins across the blood-brain barrier.
AuthorsSpencer BJ, Verma IM
JournalProc Natl Acad Sci U S A
PubMed ID17463083
Treatment of many neuronal degenerative disorders will require delivery of a therapeutic protein to neurons or glial cells across the whole CNS. The presence of the blood-brain barrier hampers the delivery of these proteins from the blood, thus necessitating a new method for delivery. Receptors on the blood-brain barrier bind ... More