Hybridization-induced dequenching of fluorescein-labeled oligonucleotides: a novel strategy for PCR detection and genotyping.
AuthorsVaughn CP,Elenitoba-Johnson KS
JournalThe American journal of pathology
PubMed ID12819008
Fluorescence-based detection methods are being increasingly utilized in molecular analyses. Sequence-specific fluorescently-labeled probes are favored because they provide specific product identification. The most established fluorescence-based detection systems employ a resonance energy transfer mechanism effected through the interaction of two or more fluorophores or functional groups conjugated to oligonucleotide probes. The ... More
Cloning and functional characterisation of the mouse P2X7 receptor.
AuthorsChessell IP, Simon J, Hibell AD, Michel AD, Barnard EA, Humphrey PP
JournalFEBS Lett
PubMed ID9849870
We have isolated a 1785-bp complementary DNA (cDNA) encoding the murine P2X7 receptor subunit from NTW8 mouse microglial cells. The encoded protein has 80%, and 85% homology to the human and rat P2X7 subunits, respectively. Functional properties of the heterologously expressed murine P2X7 homomeric receptor broadly resembled those of the ... More
Quantitation of polymerase chain reaction products by capillary electrophoresis using laser fluorescence.
AuthorsButler JM, McCord BR, Jung JM, Wilson MR, Budowle B, Allen RO
JournalJ Chromatogr B Biomed Appl
PubMed ID7820255
In samples where the amount of DNA is limited, the polymerase chain reaction (PCR) can amplify specific regions of the DNA. A quantitative analysis of the PCR product would be desirable to ensure sufficient DNA is available for analysis. In this study, we examine the use of capillary electrophoresis (CE) ... More
Prevalence of bacteria of division TM7 in human subgingival plaque and their association with disease.
AuthorsBrinig MM, Lepp PW, Ouverney CC, Armitage GC, Relman DA
JournalAppl Environ Microbiol
PubMed ID12620860
Members of the uncultivated bacterial division TM7 have been detected in the human mouth, but little information is available regarding their prevalence and diversity at this site. Human subgingival plaque samples from healthy sites and sites exhibiting various stages of periodontal disease were analyzed for the presence of TM7 bacteria. ... More
Cutting edge: a natural P451L mutation in the cytoplasmic domain impairs the function of the mouse P2X7 receptor.
AuthorsAdriouch S, Dox C, Welge V, Seman M, Koch-Nolte F, Haag F
JournalJ Immunol
PubMed ID12370338
The P2X7 receptor (P2X(7)R) is an ATP-gated channel that mediates apoptosis of cells of the immune system. The capacity of P2X(7)R to form large pores depends on its large cytoplasmic tail, which harbors a putative TNFR-related death domain. Previous transfection studies indicated that mouse P2X(7)R forms pores much less efficiently ... More
Secretion of intracellular IL-1 receptor antagonist (type 1) is dependent on P2X7 receptor activation.
AuthorsWilson HL, Francis SE, Dower SK, Crossman DC
JournalJ Immunol
PubMed ID15240711
Inflammatory mechanisms are critical in the arterial response to injury. Both IL-1 and the naturally occurring inhibitor of IL-1, IL-1R antagonist (IL-1ra), are expressed in the arterial wall, and in particular in the endothelium. Previous studies suggest that endothelial cells only make the intracellular type I isoform of IL-1ra (icIL-1ra1), ... More
An Arg307 to Gln polymorphism within the ATP-binding site causes loss of function of the human P2X7 receptor.
The P2X(7) receptor is a ligand-gated channel that is highly expressed on mononuclear cells of the immune system and that mediates ATP-induced apoptosis. Wide variations in the function of the P2X receptor have been observed, explained in part by (7)loss-of-function polymorphisms that change Glu(496) to Ala (E496A) and Ile(568) to ... More
Contribution of membrane localization to the apoptotic activity of PUMA.
AuthorsYee KS, Vousden KH,
JournalApoptosis
PubMed ID17968660
'The BH3-only protein PUMA plays an important role in the activation of apoptosis in response to p53. In different studies, PUMA has been described to function by either directly activating the pro-apoptotic proteins Bax and Bak, or by neutralizing anti-apoptotic members of the Bcl2 family. We have examined the contribution ... More
Active caspase-8 translocates into the nucleus of apoptotic cells to inactivate poly(ADP-ribose) polymerase-2.
AuthorsBenchoua A, Couriaud C, Guégan C, Tartier L, Couvert P, Friocourt G, Chelly J, Ménissier-de Murcia J, Onténiente B
JournalJ Biol Chem
PubMed ID12065591
'Caspase-8 is the prototypic initiator of the death domain receptor pathway of apoptosis. Here, we report that caspase-8 not only triggers and amplifies the apoptotic process at cytoplasmic sites but can also act as an executioner at nuclear levels. In a murine model of acute ischemia, caspase-8 is relocated into ... More
Candida albicans phospholipomannan promotes survival of phagocytosed yeasts through modulation of bad phosphorylation and macrophage apoptosis.
AuthorsIbata-Ombetta S, Idziorek T, Trinel PA, Poulain D, Jouault T
JournalJ Biol Chem
PubMed ID12551950
'The surface of the pathogenic yeast Candida albicans is coated with phospholipomannan (PLM), a phylogenetically unique glycolipid composed of beta-1,2-oligomannosides and phytoceramide. This study compared the specific contribution of PLM to the modulation of signaling pathways linked to the survival of C. albicans in macrophages in contrast to Saccharomyces cerevisiae. ... More
Involvement of extracellular signal-regulated kinase module in HIV-mediated CD4 signals controlling activation of nuclear factor-kappa B and AP-1 transcription factors.
AuthorsBriant L, Robert-Hebmann V, Sivan V, Brunet A, Pouysségur J, Devaux C
JournalJ Immunol
PubMed ID9469449
'Although the molecular mechanisms by which the HIV-1 triggers either T cell activation, anergy, or apoptosis remain poorly understood, it is well established that the interaction of HIV-1 envelope glycoproteins with cell surface CD4 delivers signals to the target cell, resulting in activation of transcription factors such as NF-kappa B ... 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
Pathogenesis of mucous cell metaplasia in a murine asthma model.
'Increased mucus production in asthma is an important cause of airflow obstruction during severe exacerbations. To better understand the changes in airway epithelium that lead to increased mucus production, ovalbumin-sensitized and -challenged mice were used. The phenotype of the epithelium was dramatically altered, resulting in increased numbers of mucous cells, ... More
A rapid and sensitive fluorometric screening assay using YO-PRO-1 to quantify tumour cell invasion through Matrigel.
AuthorsGohla A, Eckert K, Maurer HR
JournalClin Exp Metastasis
PubMed ID8871539
'A new quantitative assay for the study of tumour cell invasion in vitro is described. Employing the novel fluorescent dye YO-PRO-1, cells that penetrate Matrigel-coated transwells are counted on the basis of dye-bound cellular nucleic acid content. Following transmigration, the cells in the lower compartments are lysed by freezing in ... More
Application of digital image analysis and flow cytometry to enumerate marine viruses stained with SYBR gold.
AuthorsChen F, Lu JR, Binder BJ, Liu YC, Hodson RE
JournalAppl Environ Microbiol
PubMed ID11157214
'A novel nucleic acid stain, SYBR Gold, was used to stain marine viral particles in various types of samples. Viral particles stained with SYBR Gold yielded bright and stable fluorescent signals that could be detected by a cooled charge-coupled device camera or by flow cytometry. The fluorescent signal strength of ... More
Spectroscopic studies of single-stranded DNA ligands and oxazole yellow dyes.
AuthorsAbramo KH, Pitner JB, McGown LB
JournalBiospectroscopy
PubMed ID9547012
'Interactions between short single-stranded DNA ligands and fluorescent DNA indicator dyes were used to investigate binding selectivity of the ligands. Conformational differences among four DNA ligands of different sequence and structure, including two that form a G-quartet and two that do not, were confirmed by circular dichroism spectroscopy. Their interactions ... More
Fast separation of oligonucleotide and triplet repeat DNA on a microfabricated capillary electrophoresis device and capillary electrophoresis.
AuthorsUeda M, Kiba Y, Abe H, Arai A, Nakanishi H, Baba Y
JournalElectrophoresis
PubMed ID10634485
'A laser-induced fluorescence detection system coupled with a highly sensitive silicon-intensified target (SIT) camera is successfully applied to the imaging of a band for DNA fragment labeling by fluorescence dye in a microchannel, and to the visualizing of the separation process on a microfabricated chip. We demonstrated that an only ... More
YOPRO-1 permits cytofluorometric analysis of programmed cell death (apoptosis) without interfering with cell viability.
AuthorsIdziorek T, Estaquier J, De Bels F, Ameisen JC
JournalJ Immunol Methods
PubMed ID7561136
'In the absence of cell permeabilization, the impermeant nuclear dye YOPRO-1 permits accurate analysis of apoptosis using cytofluorometry or fluorescent microscopy. Several immune cell populations were studied including dexamethasone-treated thymocytes, irradiated peripheral blood mononuclear cells and a growth factor-depleted tumor B cell line. Excellent correlation values were found with acridine ... More
Maitotoxin-induced membrane blebbing and cell death in bovine aortic endothelial cells.
AuthorsEstacion M, Schilling WP
JournalBMC Physiol
PubMed ID11231888
'BACKGROUND: Maitotoxin, a potent cytolytic agent, causes an increase in cytosolic free Ca2+ concentration ([Ca2+]i) via activation of Ca2+-permeable, non-selective cation channels (CaNSC). Channel activation is followed by formation of large endogenous pores that allow ethidium and propidium-based vital dyes to enter the cell. Although activation of these cytolytic/oncotic pores, ... More
Stability studies of FhuA, a two-domain outer membrane protein from Escherichia coli.
AuthorsBonhivers M, Desmadril M, Moeck GS, Boulanger P, Colomer-Pallas A, Letellier L
JournalBiochemistry
PubMed ID11327884
'FhuA (MM 78.9 kDa) is an Escherichia coli outer membrane protein that transports iron coupled to ferrichrome and is the receptor for a number of bacteriophages and protein antibiotics. Its three-dimensional structure consists of a 22-stranded beta-barrel lodged in the membrane, extracellular hydrophilic loops, and a globular domain (the "cork") ... More
Focal adhesion kinase gene silencing promotes anoikis and suppresses metastasis of human pancreatic adenocarcinoma cells.
AuthorsDuxbury MS, Ito H, Zinner MJ, Ashley SW, Whang EE
JournalSurgery
PubMed ID15118593
'BACKGROUND: Inadequate or inappropriate cell-substrate contact triggers a subset of apoptotic cell death, termed anoikis. Resistance to anoikis is a characteristic of malignant cells that is associated with increased tumorigenesis and metastasis. Focal adhesion kinase (FAK) is an important regulator of cell survival and migration and cell cycle progression. We ... More
A rapid in vitro assay for evaluation of metabolism-dependent cytotoxicity of antiepileptic drugs on isolated human lymphocytes.
'In vitro assessment of human lymphocyte viability by trypan blue dye exclusion in the presence of an external metabolizing system (microsomes plus NADPH) has been shown to be a useful method in assessing predisposition to idiopathic toxicity in response to various anticonvulsant drugs. The trypan blue method, however, is labor ... More
Analysis of plasma membrane integrity by fluorescent detection of Tl(+) uptake.
AuthorsBowman AM, Nesin OM, Pakhomova ON, Pakhomov AG,
JournalJ Membr Biol
PubMed ID20623351
'The exclusion of polar dyes by healthy cells is widely employed as a simple and reliable test for cell membrane integrity. However, commonly used dyes (propidium, Yo-Pro-1, trypan blue) cannot detect membrane defects which are smaller than the dye molecule itself, such as nanopores that form by exposure to ultrashort ... More
Spectral measurements of intercalated PCR-amplified short tandem repeat alleles.
AuthorsMarino MA, Devaney JM, Davis PA, Smith JK, Girard JE
JournalAnal Chem
PubMed ID9823710
'Short tandem repeat (STR) alleles are popular for use as forensic markers due to their highly polymorphic nature. Commonly they are separated by gel electrophoresis and visualized using intercalation dyes. The purpose of this study was to determine the changes in absorbance and fluorescence of DNA-intercalation dye complexes as a ... 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
Identification and characterization of an endogenous P2X7 (P2Z) receptor in CHO-K1 cells.
AuthorsMichel AD, Chessell IP, Hibell AD, Simon J, Humphrey PP
JournalBr J Pharmacol
PubMed ID9863647
'CHO-K1 cells were examined for their cellular responses to the P2 receptor agonist, 2''- and 3''-O-(4-benzoylbenzoyl)-ATP (DbATP), and for the presence of mRNA for P2X receptors. Reverse transcriptase-polymerase chain reactions, using primers directed against the rat P2X subunits, detected the presence of P2X7 but not P2X1-P2X6 subunits. DbATP (EC50 approximately ... More
Brilliant blue G selectively blocks ATP-gated rat P2X(7) receptors.
AuthorsJiang LH, Mackenzie AB, North RA, Surprenant A
JournalMol Pharmacol
PubMed ID10860929
'There are few antagonists selective for subtypes of the several P2X receptors, but these are needed to identify the receptors expressed on native cells and tissues. In particular, P2X(4) and P2X(7) receptor subunits are colocalized on immune, epithelial, and exocrine gland cells, but both are relatively insensitive to suramin and ... More
Maitotoxin activates a nonselective cation channel and a P2Z/P2X(7)-like cytolytic pore in human skin fibroblasts.
AuthorsSchilling WP, Sinkins WG, Estacion M
JournalAm J Physiol
PubMed ID10516106
'Maitotoxin (MTX), a potent cytolytic agent, activates Ca(2+) entry via nonselective cation channels in virtually all types of cells. The identity of the channels involved and the biochemical events leading to cell lysis remain unknown. In the present study, the effect of MTX on plasmalemmal permeability of human skin fibroblasts ... More
Molecular characterization of the cytotoxic mechanism of multiwall carbon nanotubes and nano-onions on human skin fibroblast.
'The increasing use of nanotechnology in consumer products and medical applications underlies the importance of understanding its potential toxic effects to people and the environment. Although both fullerene and carbon nanotubes have been demonstrated to accumulate to cytotoxic levels within organs of various animal models and cell types and carbon ... More
Analysis of CMF1 reveals a bone morphogenetic protein-independent component of the cardiomyogenic pathway.
AuthorsPabón-Peña LM, Goodwin RL, Cise LJ, Bader D
JournalJ Biol Chem
PubMed ID10747923
'Disruption of the CMF1 function in anterior mesoderm inhibits cardiac myogenesis in avian embryos. In the present study, we show that CMF1 is a member of an emerging family of proteins that includes centromeric protein-F, mitosin, and LEK1. These proteins are characterized by their large size (350 kDa), dynamic subcellular ... More
The permeabilizing ATP receptor, P2X7. Cloning and expression of a human cDNA.
AuthorsRassendren F, Buell GN, Virginio C, Collo G, North RA, Surprenant A
JournalJ Biol Chem
PubMed ID9038151
'A cDNA was isolated from a human monocyte library that encodes the P2X7 receptor; the predicted protein is 80% identical to the rat receptor. Whole cell recordings were made from human embryonic kidney cells transfected with the human cDNA and from human macrophages. Brief applications (1-3 s) of ATP and ... More
Release of hypoacetylated and trimethylated histone H4 is an epigenetic marker of early apoptosis.
AuthorsBoix-Chornet M, Fraga MF, Villar-Garea A, Caballero R, Espada J, Nuñez A, Casado J, Largo C, Casal JI, Cigudosa JC, Franco L, Esteller M, Ballestar E
JournalJ Biol Chem
PubMed ID16531610
'Nuclear events such as chromatin condensation, DNA cleavage at internucleosomal sites, and histone release from chromatin are recognized as hallmarks of apoptosis. However, there is no complete understanding of the molecular events underlying these changes. It is likely that epigenetic changes such as DNA methylation and histone modifications that are ... More
TOTO and YOYO: new very bright fluorochromes for DNA content analyses by flow cytometry.
AuthorsHirons GT, Fawcett JJ, Crissman HA
JournalCytometry
PubMed ID7513274
'Flow cytometric (FCM) studies were performed on nuclei, ethanol-fixed CHO cells, and isolated human GM130 chromosomes stained with two new cyanine dyes, TOTO and YOYO. These fluorochromes, which are dimers of thiazole orange and oxazole yellow, respectively, have high quantum efficiencies and exhibit specificities for both DNA and RNA. Bound ... More
The nucleotide receptor P2X7 mediates actin reorganization and membrane blebbing in RAW 264.7 macrophages via p38 MAP kinase and Rho.
AuthorsPfeiffer ZA, Aga M, Prabhu U, Watters JJ, Hall DJ, Bertics PJ
JournalJ Leukoc Biol
PubMed ID15075366
'Extracellular nucleotides regulate macrophage function via P2X nucleotide receptors that form ligand-gated ion channels. In particular, P2X7 activation is characterized by pore formation, membrane blebbing, and cytokine release. P2X7 is also linked to mitogen-activated protein kinases (MAPK) and Rho-dependent pathways, which are known to affect cytoskeletal structure in other systems. ... More
Mutation of a dibasic amino acid motif within the C terminus of the P2X7 nucleotide receptor results in trafficking defects and impaired function.
'Activation of the P2X(7) receptor by extracellular nucleotides modulates multiple immune functions, including inflammatory mediator production, membrane fusion events, and apoptosis. Previous studies have revealed that the C terminus of this multimeric cation channel possesses a lipid-interaction motif that has been proposed to regulate receptor function. This domain is homologous ... 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
DNA typing of a polymerase chain reaction amplified D1S80/amelogenin multiplex using capillary electrophoresis and a mixed entangled polymer matrix.
AuthorsIsenberg AR, McCord BR, Koons BW, Budowle B, Allen RO
JournalElectrophoresis
PubMed ID8905268
'In this study, a technique was developed to separate by capillary electrophoresis (CE) the widely varying DNA fragment sizes produced by a multiplex polymerase chain reaction (PCR) amplification of the loci D1S80 and amelogenin. Experiments were performed to analyze different buffer systems and obtain optimal resolution for the separation. A ... More
Interaction of oxazole yellow dyes with DNA studied with hybrid optical tweezers and fluorescence microscopy.
AuthorsMurade CU, Subramaniam V, Otto C, Bennink ML,
JournalBiophys J
PubMed ID19651041
'We have integrated single molecule fluorescence microscopy imaging into an optical tweezers set-up and studied the force extension behavior of individual DNA molecules in the presence of various YOYO-1 and YO-PRO-1 concentrations. The fluorescence modality was used to record fluorescent images during the stretching and relaxation cycle. Force extension curves ... More
Proliferating or differentiating stimuli act on different lipid-dependent signaling pathways in nuclei of human leukemia cells.
'Previous results have shown that the human promyelocytic leukemia HL-60 cell line responds to either proliferating or differentiating stimuli. When these cells are induced to proliferate, protein kinase C (PKC)-beta II migrates toward the nucleus, whereas when they are exposed to differentiating agents, there is a nuclear translocation of the ... 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
Mechanisms underlying the impact of humic acids on DNA quantification by SYBR Green I and consequences for the analysis of soils and aquatic sediments.
'DNA quantification of soils and sediments is useful for the investigation of microbial communities and for the acquisition of their genomes that are exploited for the production of natural products. However, in such samples DNA quantification is impaired by humic acids (HA). Due to its lack of specificity and sensitivity, ... More
Extracellular ATP triggers IL-1 beta release by activating the purinergic P2Z receptor of human macrophages.
AuthorsFerrari D, Chiozzi P, Falzoni S, Dal Susino M, Melchiorri L, Baricordi OR, Di Virgilio F
JournalJ Immunol
PubMed ID9233643
'Extracellular ATP (ATPe) is known to cause release of processed IL-1 beta from LPS-treated macrophages and microglial cells. IL-1 beta release is fast and thought to be associated with cell death. We have reinvestigated this process to identify 1) the purinergic receptor involved; 2) the relationship to cell death; and ... More
Giant vesicles as models to study the interactions between membranes and proteins.
AuthorsFischer A, Oberholzer T, Luisi PL
JournalBiochim Biophys Acta
PubMed ID10930520
'The interaction between polypeptides and membranes is a fundamental aspect of cell biochemistry. Liposomes have been used in this context as in vitro systems to study such interactions. We present here the case of giant vesicles (GVs), which, due to their size (radius larger than 10 microns), mimic more closely ... More
Activation of presynaptic P2X7-like receptors depresses mossy fiber-CA3 synaptic transmission through p38 mitogen-activated protein kinase.
AuthorsArmstrong JN, Brust TB, Lewis RG, MacVicar BA
JournalJ Neurosci
PubMed ID12122056
'P2X(7) receptor subunits form homomeric ATP-gated, calcium-permeable cation channels. In this study, we used Western blots and immunocytochemistry to demonstrate that P2X(7) receptors are abundant on presynaptic terminals of mossy fiber synapses in the rat hippocampus. P2X(7)-immunoreactive protein was detected using a specific P2X(7) antibody in Western blots of protein ... More
bottleneck acts as a regulator of the microfilament network governing cellularization of the Drosophila embryo.
AuthorsSchejter ED, Wieschaus E
JournalCell
PubMed ID8402919
'A dynamic network of cortical microfilaments is associated with the cleavage furrow membranes during cellularization of the Drosophila embryo. A specific set of structural rearrangements in this network is required for orchestration and execution of its mechanistic roles. We describe the characterization of the gene bottleneck (bnk), mutations in which ... More
Pharmacologic properties of P(2Z)/P2X(7 )receptor characterized in murine dendritic cells: role on the induction of apoptosis.
AuthorsNihei OK, de Carvalho AC, Savino W, Alves LA
JournalBlood
PubMed ID10910915
'In the immune system, extracellular adenosine 5''-triphosphate (ATP) mediates a variety of effects mainly through activation of a particular receptor subtype, the pore-forming P(2Z)/P2X(7) purinoceptor. This purinergic receptor has been described chiefly in cells of hemopoietic origin such as T cells, thymocytes, monocytes, macrophages, and phagocytic cells of thymic reticulum. ... More
AOTF microscope for imaging with increased speed and spectral versatility.
AuthorsWachman ES, Niu W, Farkas DL
JournalBiophys J
PubMed ID9284289
'We have developed a new fluorescence microscope that addresses the spectral and speed limitations of current light microscopy instrumentation. In the present device, interference and neutral density filters normally used for fluorescence excitation and detection are replaced by acousto-optic tunable filters (AOTFs). Improvements are described, including the use of a ... More
A new and simple method to evaluate early membrane changes in frozen-thawed boar spermatozoa.
AuthorsPeña FJ, Saravia F, Johannisson A, Walgren M, Rodríguez-Martínez H
JournalInt J Androl
PubMed ID15811072
'Detection of early changes in the sperm plasma membrane during cryopreservation is of utmost importance when designing freezing protocols and has previously been studied in the pig species using annexin-V detection of phosphatidylserine translocation. In the present study we designed a new assay to detect these changes in boar spermatozoa, ... More
Regulation of P2X7-induced pore formation and cell death in pericyte-containing retinal microvessels.
'The purpose if this study was to elucidate how extracellular ATP causes cell death in the retinal microvasculature. Although ATP appears to serve as a vasoactive signal acting via P2X(7) and P2Y(4) purinoceptors, this nucleotide can kill microvascular cells of the retina. Because P2X(7) receptor activation causes transmembrane pores to ... More
Acousto-optical deflection-based laser beam scanning for fluorescence detection on multichannel electrophoretic microchips.
AuthorsHuang Z, Munro N, Hühmer AF, Landers JP
JournalAnal Chem
PubMed ID10596211
'Laser beam scanning driven by an acousto-optical deflector (AOD) is presented for multimicrochannel laser-induced fluorescence (LIF) detection during microchip-based electrophoresis. While fast laser beam scanning for LIF detection on capillary or microchannel arrays can been achieved with galvanometric scanning or a translating stage, it can also be accomplished by using ... More
Reconstitution of FhuA, an Escherichia coli outer membrane protein, into liposomes. Binding of phage T5 to Fhua triggers the transfer of DNA into the proteoliposomes.
AuthorsPlançon L, Chami M, Letellier L
JournalJ Biol Chem
PubMed ID9201994
'The Escherichia coli outer membrane protein FhuA catalyzes the transport of ferrichrome and is the receptor of bacteriophage T5. Purified FhuA was reconstituted into liposomes. The size of the proteoliposomes and the distribution of the proteins in the vesicles were determined by freeze fracture electron microscopy. Unilamellar vesicles with a ... More
Real-time observation of a single DNA digestion by lambda exonuclease under a fluorescence microscope field.
'A fluorescence microscopy technique has been developed to visualize the behavior of individual DNA and protein molecules. Real-time direct observation of a single DNA molecule can be used to investigate the dynamics of DNA-protein interactions, such as the DNA digestion reaction by lambda exonuclease. In conventional methods it is impossible ... More
Modeling ligand-gated receptor activity. FhuA-mediated ferrichrome efflux from lipid vesicles triggered by phage T5.
'An in vitro assay of iron-ferrichrome translocation across the FhuA protein of outer membranes from Escherichia coli has been devised. Upon reconstitution into large lipid vesicles, bacteriophage T5 binds to this polyvalent receptor, triggering a conformational change that resulted in channel opening. This facilitates the translocation of an iron(III)-siderophore, without ... More
Rapid analysis of the short tandem repeat HUMTH01 by capillary electrophoresis.
AuthorsButler JM, McCord BR, Jung JM, Allen RO
JournalBiotechniques
PubMed ID7873175
'Using capillary electrophoresis, we demonstrate separation and analysis of the short tandem repeat HUMTH01 in under 10 min with 3 bp resolution. Separation of the PCR products, which range in size from 179 to 203 bp, is achieved using hydroxyethyl cellulose as the separation medium and a novel single-step voltage ... More
P2X7 receptor activates multiple selective dye-permeation pathways in RAW 264.7 and human embryonic kidney 293 cells.
AuthorsCankurtaran-Sayar S, Sayar K, Ugur M,
JournalMol Pharmacol
PubMed ID19749088
'P2X7 receptor has gained an increasing importance as a drug target. One important response to P2X7 receptor stimulation is the uptake of large molecular weight tracers into cells. However, mechanism for this response is not understood clearly, but it is generally believed that a nonselective large pore protein forms this ... More
Ultrafast excited-state dynamics of oxazole yellow DNA intercalators.
AuthorsFürstenberg A, Vauthey E,
JournalJ Phys Chem B
PubMed ID17929857
'The excited-state dynamics of the DNA intercalator YO-PRO-1 and of three derivatives has been investigated in water and in DNA using ultrafast fluorescence spectroscopy. In the free form, the singly charged dyes exist both as monomers and as H-dimers, while the doubly charged dyes exist predominantly as monomers. Both forms ... More
Phosphorylation of human keratin 8 in vivo at conserved head domain serine 23 and at epidermal growth factor-stimulated tail domain serine 431.
AuthorsKu NO, Omary MB
JournalJ Biol Chem
PubMed ID9054461
'Dynamic phosphorylation is one mechanism that regulates the more than 20 keratin type I and II intermediate filament proteins in epithelial cells. The major type II keratin in "simple type" glandular epithelia is keratin 8 (K8). We used biochemical and mutational approaches to localize two major in vivo phosphorylation sites ... More
In vivo targeting of dendritic cells for activation of cellular immunity using vaccine carriers based on pH-responsive microparticles.
AuthorsKwon YJ, James E, Shastri N, Fréchet JM
JournalProc Natl Acad Sci U S A
PubMed ID16344458
'Activating the immune system to trigger a specific response is a major challenge in vaccine development. In particular, activating sufficient cytotoxic T lymphocyte-mediated cellular immunity, which is crucial for the treatment of many diseases including cancer and AIDS, has proven to be especially challenging. In this study, antigens were encapsulated ... More
Evaluation of silica resins for direct and efficient extraction of DNA from complex biological matrices in a miniaturized format.
AuthorsTian H, Hühmer AF, Landers JP
JournalAnal Biochem
PubMed ID10906238
'For DNA purification to be functionally integrated into the microchip for high-throughput DNA analysis, a miniaturized purification process must be developed that can be easily adapted to the microchip format. In this study, we evaluate the effectiveness of a variety of silica resins for miniaturized DNA purification and gauge the ... More
Analysis of internucleosomal DNA fragmentation in apoptotic thymocytes by dynamic sieving capillary electrophoresis.
AuthorsEvans MD, Wolfe JT, Perrett D, Lunec J, Herbert KE
JournalJ Chromatogr A
PubMed ID7767461
'The analysis, by slab gel electrophoresis, of internucleosomal DNA cleavage or laddering, characteristic of apoptosis in many cell systems, is labour intensive, difficult to automate and at best only semi-quantitative. In this report we show that CE, using dilute solutions of hydroxyethylcellulose as a replaceable sieving matrix, can be applied ... More
A single site on the epsilon subunit is responsible for the change in ACh receptor channel conductance during skeletal muscle development.
AuthorsMurray N, Zheng YC, Mandel G, Brehm P, Bolinger R, Reuer Q, Kullberg R
JournalNeuron
PubMed ID7718247
'Four critically positioned amino acids on each of the alpha, beta, delta, and gamma subunits of the Torpedo nicotinic acetylcholine receptor are determinants of channel conductance. Our results show that the gamma and epsilon subunits of Xenopus muscle receptors are identical at all four positions, despite the fact that alpha ... More
Analysis of a variable number of tandem repeats in a heart disease gene by capillary electrophoresis with laser-induced fluorescence detector.
AuthorsNishimura A, Tsuhako M, Miki T, Ogihara T, Baba Y
JournalChem Pharm Bull (Tokyo)
PubMed ID9501464
'Capillary electrophoresis with laser-induced fluorescence detector (CE-LIF) was successfully applied to the analysis of the variable number of tandem repeats (VNTR) in a human apolipoprotein B gene (APOB). Apolipoprotein B VNTR alleles containing more than 35 repeat units are a significant risk factor for heart disease. Thus, we developed a ... More
In situ analyses of genome instability in breast cancer.
AuthorsChin K, de Solorzano CO, Knowles D, Jones A, Chou W, Rodriguez EG, Kuo WL, Ljung BM, Chew K, Myambo K, Miranda M, Krig S, Garbe J, Stampfer M, Yaswen P, Gray JW, Lockett SJ
JournalNat Genet
PubMed ID15300252
'Transition through telomere crisis is thought to be a crucial event in the development of most breast carcinomas. Our goal in this study was to determine where this occurs in the context of histologically defined breast cancer progression. To this end, we assessed genome instability (using fluorescence in situ hybridization) ... More
Virus-assisted loading of polymer nanocontainer.
AuthorsGraff A, Sauer M, Van Gelder P, Meier W
JournalProc Natl Acad Sci U S A
PubMed ID11917114
'We present a DNA-containing polymeric nanocontainer using the self-assembled superstructure of amphiphilic block copolymers in aqueous solutions. To demonstrate that DNA translocation is possible across a completely synthetic block copolymer membrane, we have used a phage transfection strategy as a DNA-transfer model system. For this purpose the bacterial channel forming ... More
Photobleaching of asymmetric cyanines used for fluorescence imaging of single DNA molecules.
AuthorsKanony C, Akerman B, Tuite E
JournalJ Am Chem Soc
PubMed ID11506554
'The photobleaching of the cyanine dyes YO and YOYO has been investigated for both free and DNA-bound dyes, using absorption and fluorescence spectroscopy coupled with fluorescence microscopy. For the free dyes, the nature of the reactive species involved in the photodegradation process is different for the monomer and the dimer, ... More
Flow cytometry-based apoptosis detection.
AuthorsWlodkowic D, Skommer J, Darzynkiewicz Z,
JournalMethods Mol Biol
PubMed ID19609746
'An apoptosing cell demonstrates multitude of characteristic morphological and biochemical features, which vary depending on the stimuli and the cell type. The gross majority of classical apoptotic hallmarks can be rapidly examined by flow and image cytometry. Cytometry thus became a technology of choice in diverse studies of cellular demise. ... More
Staining of DNA in live and fixed cells.
AuthorsCrissman HA, Hirons GT
JournalMethods Cell Biol
PubMed ID7532262
The nucleic acid ligand. A new tool for molecular recognition.
AuthorsMcGown LB, Joseph MJ, Pitner JB, Vonk GP, Linn CP
JournalAnal Chem
PubMed ID8633764
Single-cell enumeration of an uncultivated TM7 subgroup in the human subgingival crevice.
AuthorsOuverney CC, Armitage GC, Relman DA
JournalAppl Environ Microbiol
PubMed ID14532094
Specific oligonucleotide hybridization conditions were established for single-cell enumeration of uncultivated TM7 and IO25 bacteria by using clones expressing heterologous 16S rRNA. In situ analysis of human subgingival crevice specimens revealed that a greater proportion of samples from sites of chronic periodontitis than from healthy sites contained TM7 subgroup IO25. ... More
Measuring apoptosis at the single cell level.
AuthorsBouchier-Hayes L, Muñoz-Pinedo C, Connell S, Green DR,
JournalMethods
PubMed ID18314052
The use of live cell microscopy has made a number of contributions to the study of apoptosis. Many of the tools and techniques are available that allow us to image the key events that occur during cell death including mitochondrial outer membrane permeabilization, mitochondrial transmembrane potential changes, translocation of Bcl-2 ... More
Homogeneous quantitative assay of hepatitis C virus RNA by polymerase chain reaction in the presence of a fluorescent intercalater.
We have developed a homogeneous quantitative assay of DNA/RNA by performing PCR in the presence of an oxazole yellow derivative, a fluorescent DNA intercalative dye, and monitoring the fluorescence intensity of the PCR reaction mixture during PCR cycles. We have demonstrated the applicability of this assay by use it to ... More
Pharmaceutical-induced cell apoptosis characterized by capillary zone electrophoresis.
AuthorsLiu BF, Zhang L, Liu HH, Lu YT
JournalAnal Biochem
PubMed ID11567522
Capillary zone electrophoresis (CZE) with a non-gel-sieving system was employed to characterize actinomycin D-induced cell apoptosis by measuring cellular DNA damage, termed DNA ladder, which proved to be thoroughly different from the DNA damage pattern of cell necrosis. The results by CZE analyses were identical to those obtained by conventional ... More
Wide geographic distribution of bacteriophages that lyse the same indigenous freshwater isolate (Sphingomonas sp. strain B18).
AuthorsWolf A, Wiese J, Jost G, Witzel KP
JournalAppl Environ Microbiol
PubMed ID12676728
An indigenous freshwater bacterium (Sphingomonas sp. strain B18) from Lake Plubetasee (Schleswig-Holstein, Germany) was used to isolate 44 phages from 13 very different freshwater and brackish habitats in distant geographic areas. This bacterial strain was very sensitive to a broad spectrum of phages from different aquatic environments. Phages isolated from ... More
Cytokinesis in the Arabidopsis embryo involves the syntaxin-related KNOLLE gene product.
AuthorsLukowitz W, Mayer U, Jürgens G
JournalCell
PubMed ID8548827
The embryo of the flowering plant Arabidopsis develops by a regular pattern of cell divisions and cell shape changes. Mutations in the KNOLLE (KN) gene affect the rate and plane of cell divisions as well as cell morphology, resulting in mutant seedlings with a disturbed radial organization of tissue layers. ... More
Neuronal P2X transmitter-gated cation channels change their ion selectivity in seconds.
AuthorsKhakh BS, Bao XR, Labarca C, Lester HA
JournalNat Neurosci
PubMed ID10204538
Fast synaptic transmission depends on the selective ionic permeability of transmitter-gated ion channels. Here we show changes in the ion selectivity of neuronal P2X transmitter-gated cation channels as a function of time (on the order of seconds) and previous ATP exposure. Heterologously expressed P2X2, P2X2/P2X3 and P2X4 channels as well ... More
Isolation, structure elucidation and cytotoxic evaluation of endiandrin B from the Australian rainforest plant Endiandra anthropophagorum.
Chemical investigations of the DCM extract from the roots of Endiandra anthropophagorum resulted in the isolation of a new cyclobutane lignan endiandrin B (1), together with the known natural products, endiandrin A (2), and (-)-dihydroguaiaretic acid (3). The structure of 1 was determined by extensive spectroscopic analyses, and confirmed by ... More
Pore dilation of neuronal P2X receptor channels.
AuthorsVirginio C, MacKenzie A, Rassendren FA, North RA, Surprenant A
JournalNat Neurosci
PubMed ID10204537
P2X receptors are ligand-gated ion channels activated by the binding of extracellular adenosine 5'-triphosphate (ATP). Brief (< 1 s) applications of ATP to nodose ganglion neurons or to cells transfected with P2X2 or P2X4 receptor cDNAs induce the opening of a channel selectively permeable to small cations within milliseconds. We ... More
P2X7 purinoceptor expression in Xenopus oocytes is not sufficient to produce a pore-forming P2Z-like phenotype.
AuthorsPetrou S, Ugur M, Drummond RM, Singer JJ, Walsh JV
JournalFEBS Lett
PubMed ID9271232
The purinergic rP2X7 receptor expressed in a number of heterologous systems not only functions as a cation channel but also gives rise to a P2Z-like response, i.e. a reversible membrane permeabilization that allows the passage of molecules with molecular masses of > or = 300 Da. We investigated the properties ... More
DNA melting in the presence of fluorescent intercalating oxazole yellow dyes measured with a gel-based assay.
AuthorsBjorndal MT, Fygenson DK
JournalBiopolymers
PubMed ID12209471
We measured the effect of the intercalating oxazole yellow DNA dye quinolinium,4-[(3-methyl-2(3H)-benzoxazolylidene)methyl]-1-[3-(trimethylammonio)propyl]-,diiodide (YO-PRO) and its homodimer (YOYO) on the melting of self-complementary DNA duplexes using a gel-based assay. The assay, which requires a self-complementary DNA sequence, is independent of the optical properties of the molecules in solution. The melting temperature ... More
The comet assay is a single cell gel electrophoresis test currently used as a qualitative and quantitative genotoxicity test. However, some of the results from this comet assay and current knowledge on apoptosis lead us to suspect the presence of some false positive results. The aim of this study was ... More
Visualization of a specific sequence on a single large DNA molecule using fluorescence microscopy based on a new DNA-stretching method.
AuthorsOana H, Ueda M, Washizu M
JournalBiochem Biophys Res Commun
PubMed ID10548504
A method for analyzing large DNA which makes it possible to obtain spatial information on the positions of specific sequences along a DNA molecule has been developed. Making use of the fact that large DNA molecules are stably elongated under an alternating-current field in a concentrated linear polymer solution, the ... 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
Lateral elements inside synaptonemal complex-like polycomplexes in ndt80 mutants of yeast bind DNA.
AuthorsBhuiyan H, Dahlfors G, Schmekel K
JournalGenetics
PubMed ID12618393
The synaptonemal complex (SC) keeps the synapsed homologous chromosomes together during pachytene in meiotic prophase I. Structures that resemble stacks of SCs, polycomplexes, are sometimes found before or after pachytene. We have investigated ndt80 mutants of yeast, which arrest in pachytene. SCs appear normal in spread chromosome preparations, but are ... More
Single strand conformation polymorphism analysis of ras oncogene by capillary electrophoresis with laser-induced fluorescence detector.
AuthorsNishimura A, Tsuhako M
JournalChem Pharm Bull (Tokyo)
PubMed ID10866135
Single strand conformation polymorphism (SSCP) analysis of the N-ras oncogene was achieved by capillary electrophoresis with a laser-induced fluorescence detector (CE-LIF) using methylcellulose as a molecular sieving agent. The PCR-amplified N-ras oncogene, which is known to have a point mutation at codon 61 in the neuroblastoma, was investigated by CE-LIF ... More
Dynamic changes in the TRPA1 selectivity filter lead to progressive but reversible pore dilation.
AuthorsBanke TG, Chaplan SR, Wickenden AD,
JournalAm J Physiol Cell Physiol
PubMed ID20457836
TRPA1 is a nonselective cation channel belonging to the transient receptor potential (TRP) family that is expressed in peripheral sensory neurons and may play important roles in pain perception and inflammation. We found that agonist stimulation of TRPA1, along with other members of the TRP family (TRPV1-4 and TRPM8), can ... More
Chlorophenols and chlorocatechols induce apoptosis in human lymphocytes (in vitro).
AuthorsMichalowicz J, Sicinska P,
JournalToxicol Lett
PubMed ID19766705
In this work the effect of 2,4,5-trichlorophenol (2,4,5-TCP), pentachlorophenol (PCP), 4,6-dichloroguaiacol (4,6-DCG), tetrachloroguaiacol (TeCG), 4,5-dichlorocatechol (4,5-DCC) and tetrachlorocatechol (TeCC) on the induction of apoptosis in human peripheral blood lymphocytes was examined. The analysis of the changes in mitochondrial transmembrane potential (DeltaPsi(m)) was performed using JC-9 fluorescent probe. It was noted ... More
The polarization of fluorescence of DNA stains depends on the incorporation density of the dye molecules.
AuthorsUy JL, Asbury CL, Petersen TW, van den Engh G,
JournalCytometry A
PubMed ID15351985
BACKGROUND: The fluorescence induced by polarized light sources, such as the lasers that are used in flow cytometry, is often polarized and anisotropic. In addition, most optical detector systems are sensitive to the direction of polarization. These two factors influence the accuracy of fluorescence intensity measurements. The intensity of two ... More
Electroporating fields target oxidatively damaged areas in the cell membrane.
Reversible electropermeabilization (electroporation) is widely used to facilitate the introduction of genetic material and pharmaceutical agents into living cells. Although considerable knowledge has been gained from the study of real and simulated model membranes in electric fields, efforts to optimize electroporation protocols are limited by a lack of detailed understanding ... More
Vascular endothelial growth factor-C protects prostate cancer cells from oxidative stress by the activation of mammalian target of rapamycin complex-2 and AKT-1.
AuthorsMuders MH, Zhang H, Wang E, Tindall DJ, Datta K,
JournalCancer Res
PubMed ID19638584
Recurrence and subsequent metastatic transformation of cancer develops from a subset of malignant cells, which show the ability to resist stress and to adopt to a changing microenvironment. These tumor cells have distinctly different growth factor pathways and antiapoptotic responses compared with the vast majority of cancer cells. Long-term therapeutic ... More
Comparison of apoptosis and mortality measurements in peripheral blood mononuclear cells (PBMCs) using multiple methods.
Death through apoptosis is the main process by which aged cells that have lost their function are eliminated. Apoptotic cells are usually detected microscopically by changes in their morphology. However, determination of early apoptotic events is important for in vitro (and ex vivo) studies. The main objective of the present ... 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
ATP-induced P2X7-associated uptake of large molecules involves distinct mechanisms for cations and anions in macrophages.
AuthorsSchachter J, Motta AP, de Souza Zamorano A, da Silva-Souza HA, Guimarães MZ, Persechini PM,
JournalJ Cell Sci
PubMed ID18782864
Macrophages express the P2X(7) receptor and other nucleotide (P2) receptors, and display the phenomenon of extracellular ATP (ATP(e))-induced P2X(7)-dependent membrane permeabilization, which occurs through a poorly understood mechanism. We used patch-clamp recordings, cytoplasmic Ca(2+) measurements and fluorescent dye uptake assays to compare P2X(7)-associated transport phenomena of macrophages and HEK-293 cells ... More
Rapid mitochondrial DNA typing using restriction enzyme digestion of polymerase chain reaction amplicons followed by capillary electrophoresis separation with laser-induced fluorescence detection.
AuthorsButler JM, Wilson MR, Reeder DJ
JournalElectrophoresis
PubMed ID9511872
The polymorphic control region of mitochondrial DNA (mtDNA) is becoming more commonly used in forensic applications to differentiate among individuals in a population. Two hypervariable regions (HV1 and HV2) are often sequenced following amplification of the mtDNA via the polymerase chain reaction (PCR). More rapid screening assays would reduce both ... More
Confocal restricted-height imaging of suspension cells (CRISC) in a PDMS microdevice during apoptosis.
AuthorsMuñoz-Pinedo C, Green DR, van den Berg A
JournalLab Chip
PubMed ID15915255
We have monitored and imaged cell death induced in human leukemic U937 cells over time using three-color confocal imaging. Three different apoptotic inducers, anti-Fas, TNF-alpha and Etoposide were used. Individual cascaded events such as loss of mitochondrial transmembrane potential, exposure of phosphatidyl-serine, membrane blebbing and permeabilization of the cell membrane ... More
Detection of dead cells and measurement of cell killing by flow cytometry.
AuthorsKing MA
JournalJ Immunol Methods
PubMed ID10986413
A flow cytometer can quickly perform numerous quantitative, sensitive measurements on each individual cell within a large, heterogeneous population. The modern commercially available analytical instruments, which can be found in most hospitals, pathology laboratories, and cell biology research laboratories in the industrially developed countries, can now routinely measure fluorescence simultaneously ... More
Direct measurement of conformational changes on DNA molecule intercalating with a fluorescence dye in an electrophoretic buffer solution by means of atomic force microscopy.
AuthorsKaji N, Ueda M, Baba Y
JournalElectrophoresis
PubMed ID11669510
By means of atomic force microscopy (AFM), we performed the direct imaging of DNA molecules (200, 500, 1000 bp) in a Tris-borate buffer solution, and measured the contour length and the end-to-end distance of DNA. Processing the data according to the worm-like chain model, we calculated the persistence length of ... More
Adenovirus E3-6.7K maintains calcium homeostasis and prevents apoptosis and arachidonic acid release.
AuthorsMoise AR, Grant JR, Vitalis TZ, Jefferies WA
JournalJ Virol
PubMed ID11799152
E3-6.7K is a small and hydrophobic membrane glycoprotein encoded by the E3 region of subgroup C adenovirus. Recently, E3-6.7K has been shown to be required for the downregulation of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) receptors by the adenovirus E3/10.4K and E3/14.5K complex of proteins. We demonstrate here that ... More
An homogeneous fluorescence polymerase chain reaction assay to identify Salmonella.
AuthorsTseng SY, Macool D, Elliott V, Tice G, Jackson R, Barbour M, Amorese D
JournalAnal Biochem
PubMed ID9056213
We have developed a semiquantitative homogeneous fluorescence assay that combines polymerase chain reaction (PCR) amplification with direct fluorescence detection (HF-PCR). The assay eliminates the need to perform gel electrophoresis on test samples. Using a set of Salmonella-specific primers, this system was used to verify suspect colonies from culture plates as ... More
Direct gene transfer into rat liver cells by in vivo electroporation.
AuthorsSuzuki T, Shin BC, Fujikura K, Matsuzaki T, Takata K
JournalFEBS Lett
PubMed ID9563509
In vivo electro-transfection efficiency and manner of transferred gene expression were investigated by fluorescence microscopic image analysis. Green fluorescent protein (GFP) gene was used as the genetic marker. Electroporation was carried out on the liver of live rats by use of disk electrodes mounted in the tips of tweezers, which ... More
The P2X7 nucleotide receptor mediates skeletal mechanotransduction.
AuthorsLi J, Liu D, Ke HZ, Duncan RL, Turner CH
JournalJ Biol Chem
PubMed ID16269410
The P2X7 nucleotide receptor (P2X7R) is an ATP-gated ion channel expressed in many cell types including osteoblasts and osteocytes. Mice with a null mutation of P2X7R have osteopenia in load bearing bones, suggesting that the P2X7R may be involved in the skeletal response to mechanical loading. We found the skeletal ... More