Phycobilisomes of Porphyridium cruentum: pigment analysis.
AuthorsGantt E, Lipschultz CA
JournalBiochemistry
PubMed ID4407620
Mixed skin cell-lymphocyte culture reaction (MSLR) in man as an vitro model of lympho-epidermal interactions: FACS analysis of epidermal cell and T-cell involved subpopulations.
AuthorsCzernielewski JM, Vaigot P, Schmitt D, Prunieras M
JournalBr J Dermatol
PubMed ID6234924
Antioxidant properties of isotorachrysone isolated from Rhamnus nakaharai.
AuthorsHsiao G, Ko FN, Lin CN, Teng CM
JournalBiochim Biophys Acta
PubMed ID8948496
'Isotorachrysone inhibited iron-induced lipid peroxidation with an IC50 value of 1.64 +/- 0.08 microM in rat brain homogenates, and was comparable in potency to butylated hydroxytoluene and was more potent than alpha-tocopherol or desferrioxamine. The mechanism of antioxidant properties were then examined. Isotorachrysone could scavenge the stable free radical diphenyl-p-picrylhydrazyl. ... More
Alpha interferon accelerates lateral diffusion of Daudi cell surface differentiation antigens: measurement by fluorescence redistribution after photobleaching.
AuthorsBalint E, Aszalos A, Grimley PM
JournalBiochem Biophys Res Commun
PubMed ID2462420
'Lateral diffusion coefficients (D) of two surface differentiation antigens (sIgM and Bp35) were determined on interferon-sensitive (-IFs) or resistant (-IFr) Daudi cells by fluorescence photobleaching, using monospecific FITC-anti-IgM or PE-anti-Leu 16 probes. For untreated Daudi -IFs, mean (D) were 5.8 and 5.3 (x10(-10) cm2/sec). These increased, to 11 and 7.9 ... More
Antioxidant properties of steroids.
AuthorsMooradian AD
JournalJ Steroid Biochem Mol Biol
PubMed ID8518206
'To determine the relative ranking of antioxidative potential of various steroids the effect of 14 steroid compounds on the fluorescence of phycoerythrin was monitored over time following the addition of a peroxy radical generator 2,2''-azo-bis (2-amidino-propane) dihydrochloride. The rate of decay of fluorescence in the presence of a 200 nM ... More
Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements.
AuthorsPrior RL, Wu X, Schaich K,
JournalJ Agric Food Chem
PubMed ID15884874
'Methods available for the measurement of antioxidant capacity are reviewed, presenting the general chemistry underlying the assays, the types of molecules detected, and the most important advantages and shortcomings of each method. This overview provides a basis and rationale for developing standardized antioxidant capacity methods for the food, nutraceutical, and ... More
Isolation, crystallization, crystal structure analysis and refinement of B-phycoerythrin from the red alga Porphyridium sordidum at 2.2 A resolution.
AuthorsFicner R, Lobeck K, Schmidt G, Huber R
JournalJ Mol Biol
PubMed ID1469725
'The light-harvesting pigment-protein complex B-phycoerythrin from the red alga Porphyridium sordidum has been isolated and crystallized. B-Phycoerythrin consists of three different subunits forming an (alpha beta)6 gamma aggregate. The three-dimensional structure of the (alpha beta)6 hexamer was solved by Patterson search techniques using the molecular model of C-phycocyanin from Fremyella ... More
[Immunophenotyping of lymphocyte subpopulations using monoclonal antibodies in an analytical flow cytometer: evaluation of the method and reference values]
AuthorsNeumeier D, Pauls R, Wifling I, Fateh-Moghadam A, Knedel M
JournalJ Clin Chem Clin Biochem
PubMed ID4086981
'Analytical flow cytometers which are easy to operate, and monoclonal antibodies specific for cell surface determinants of lymphocytes, offer a simple and rapid method for the identification of the immunological phenotype of lymphocyte subsets. We investigated mononuclear cells isolated from EDTA-blood bv density gradient centrifugation. The intraassay precision (n = ... More
The antioxidative potential of cerebral microvessels in experimental diabetes mellitus.
AuthorsMooradian AD
JournalBrain Res
PubMed ID7728529
'To determine if experimental diabetes is associated with decreased antioxidative potential along with increased peroxidation of lipids in cerebral microvessels, streptozotocin-induced diabetic rats were compared with control rats and to diabetic rats treated with insulin. Isolated cerebral microvessels from diabetic rats had significantly higher concentrations of malondialdehyde (MDA; micrograms/mg protein) ... More
Age-related changes in the antioxidative potential of cerebral microvessels.
AuthorsMooradian AD, Uko-eninn A
JournalBrain Res
PubMed ID7728528
'To determine if aging in rats is associated with increased susceptibility of cerebral microvessels to oxidative damage microvessels from the cerebrum of 4-, 12-, 18- and 26-month-old male Fischer 344 rats were studied. The malondialdehyde (MDA) (micrograms/mg protein) content of cerebral microvessels from 12-month-old rats (0.032 +/- 0.002) was significantly ... More
Three-color immunofluorescence analysis of mouse B-lymphocyte subpopulations.
AuthorsParks DR, Hardy RR, Herzenberg LA
JournalCytometry
PubMed ID6370630
'We have modified a fluorescence-activated cell sorter (FACS) to make three independent immunofluorescence measurements on each cell and used this system to study mouse B-lymphocyte subpopulations. An argon-ion laser (emitting at 488 nm) excites fluorescein- and phycoerythrin-labeled reagents, and a tunable dye laser charged with rhodamine 6G (emitting at 615 ... More
Improved detection of rare CALLA-positive cells in peripheral blood using multiparameter flow cytometry.
'A major limitation to the detection of rare cell types in the peripheral blood using monoclonal antibodies is nonspecific binding of the antibody reagent to normal cells. Detection of rare common acute lymphoblastic leukemia antigen (CALLA)-positive cells in peripheral blood is significantly improved by using multiple flow cytometric parameters to ... More
High-performance optical filters for fluorescence analysis.
AuthorsMarcus DA
JournalCell Motil Cytoskeleton
PubMed ID3180249
'Recent advances in thin film optical coating technology significantly improve the filters available for fluorescence spectroscopy. Bandpass and long- and shortpass filters with very sharply defined edges can provide from 10(-5) to 10(-6) blocking within 10-15 nm of the transmission region and are ideal for use as excitation and emission ... More
Oxidative stress in lungs of mice infected with influenza A virus.
AuthorsBuffinton GD, Christen S, Peterhans E, Stocker R
JournalFree Radic Res Commun
PubMed ID1321077
'As oxidative stress has been implicated in the pathogenesis of certain viral diseases we determined antioxidant and prooxidant parameters in lungs and bronchoalveolar lavage fluid (BALF) of mice infected with a lethal dose of influenza A/PR8/34 virus. Viral infection was characterized by massive infiltration of leukocytes, mainly polymorphonuclear leukocytes, into ... More
Two-color immunofluorescence and flow cytometry analysis of lymphocytes in long-term renal allotransplant recipients: identification of a major Leu-7+/Leu-3+ subpopulation.
AuthorsLegendre CM, Guttmann RD, Hou SK, Jean R
JournalJ Immunol
PubMed ID3924997
'Using two-color immunofluorescence with fluorescein isothiocyanate (FITC)- and phycoerythrin (PE)-labeled monoclonal antibodies to human lymphocyte antigens and flow cytometry, we studied lymphocyte subsets in 16 long-term renal allotransplant recipients at risk for a mean of 78 +/- 15 mo. The absolute number of Leu-1+, Leu-2a+, and Leu-3a+ lymphocytes is significantly ... More
Bile acids: antioxidants or enhancers of peroxidation depending on lipid concentration.
AuthorsDeLange RJ, Glazer AN
JournalArch Biochem Biophys
PubMed ID2297222
'Using three different assay systems, we have discovered a heretofore unrecognized antioxidant property of bile acids at physiological concentrations. Bile acids inhibit peroxidation of the polyunsaturated lipid, linoleic acid, and of the highly fluorescent protein phycoerythrin. In part, the antioxidant activity results from scavenging of peroxyl radicals by direct oxidation ... More
Glucocorticoid receptor binding to rat liver nuclei occurs without nuclear transport.
AuthorsMiyashita Y, Miller M, Yen PM, Harmon JM, Hanover JA, Simons SS
JournalJ Steroid Biochem Mol Biol
PubMed ID9831479
'The binding of cell-free activated glucocorticoid receptor-steroid complexes from HTC cells to various preparations of HTC and rat liver nuclei has been examined under conditions that did or did not support the nuclear translocation of macromolecules via nuclear pores. To the best of our knowledge, this is the first such ... More
Demonstration of B-cell maturation in X-linked immunodeficient mice by simultaneous three-colour immunofluorescence.
AuthorsHardy RR, Hayakawa K, Parks DR, Herzenberg LA
JournalNature
PubMed ID6358897
'CBA/N mice carrying the X-linked immune deficiency gene (xid) have fewer splenic B cells than normal CBA mice and are unresponsive to a certain class of antigens. Studies of B-cell surface-marker expression and immune responsiveness have led to the commonly accepted idea that the B cells in adult xid mice ... More
A fluorometric method for measurement of oxygen radical-scavenging activity of water-soluble antioxidants.
AuthorsNaguib YM
JournalAnal Biochem
PubMed ID10933861
'The relative activities of the antioxidants Trolox, ascorbic acid, uric acid, quercetin, and rutin, and the activities of total antioxidants in serum samples were determined using a fluorometric assay based on the dye 6-carboxyfluoroscein (6C-Fl) as a fluorescent indicator; 2,2''-azobis-2-amidinopropane hydrochloride (AAPH) as a peroxyl radical generator; 6-hydroxy-2,5,7, 8-tetramethyl-1-chroman-2-carboxylic acid ... More
Phycoerythrin fluorescence-based assay for peroxy radicals: a screen for biologically relevant protective agents.
AuthorsDeLange RJ, Glazer AN
JournalAnal Biochem
PubMed ID2567129
'Under the conditions of this assay, antioxidants that react rapidly with peroxy free radicals (e.g., ascorbate, vitamin E analogs, urate), protect phycoerythrin completely from damage by such radicals generated by thermal decomposition of 2,2''-azobis(2-amidinopropane); other compounds provide partial concentration-dependent protection. Change in phycoerythrin fluorescence emission with time provides a measure ... More
Homogeneous model immunoassay of thyroxine by phase-modulation fluorescence spectroscopy.
'We describe a homogeneous competitive model immunoassay for determination of thyroxine by multifrequency phase-modulation fluorescence. Using a nonradiative energy transfer transduction mechanism, B-phycoerythrin conjugated to thyroxine is the energy donor and a carboxymethylindocyanine dye conjugated to anti-thyroxine antibody is the energy acceptor. Energy transfer from B-phycoerythrin to the acceptor results ... More
Suppressing Brownian motion of individual biomolecules in solution.
AuthorsCohen AE, Moerner WE
JournalProc Natl Acad Sci U S A
PubMed ID16537418
'Single biomolecules in free solution have long been of interest for detailed study by optical methods, but Brownian motion prevents the observation of one single molecule for extended periods. We have used an anti-Brownian electrokinetic (ABEL) trap to trap individual protein molecules in free solution, under ambient conditions, without requiring ... More
Automated assay of oxygen radical absorbance capacity with the COBAS FARA II.
AuthorsCao G, Verdon CP, Wu AH, Wang H, Prior RL
JournalClin Chem
PubMed ID7497614
'Reactive oxygen species are of interest in biology and medicine because of evidence relating them to aging and disease processes. A relatively simple but sensitive and reliable method for quantifying the oxygen radical absorbance capacity (ORAC) of antioxidants in biological tissues has been automated for use with the COBAS FARA ... More
Subsets of T lymphocyte subpopulations after human marrow transplantation defined by monoclonal antibodies and two-color fluorescence.
Spectral properties of phycobilisomes and phycobiliproteins from the blue-green alga-nostoc sp.
AuthorsGray BH, Gantt E
JournalPhotochem Photobiol
PubMed ID806082
Phycobiliproteins.
AuthorsGlazer AN
JournalMethods Enzymol
PubMed ID3148835
Coupling of monoclonal antibodies with fluorophores.
AuthorsHaugland RP
JournalMethods Mol Biol
PubMed ID7550684
Comparative biochemistry of photosynthetic light-harvesting systems.
AuthorsGlazer AN
JournalAnnu Rev Biochem
PubMed ID6412621
Light harvesting by phycobilisomes.
AuthorsGlazer AN
JournalAnnu Rev Biophys Biophys Chem
PubMed ID3924069
The use of phycobiliproteins as fluorescent labels in immunoassay.
AuthorsKronick MN
JournalJ Immunol Methods
PubMed ID3528294
Light guides. Directional energy transfer in a photosynthetic antenna.
AuthorsGlazer AN
JournalJ Biol Chem
PubMed ID2491842
Immunoassay techniques with fluorescent phycobiliprotein conjugates.
AuthorsKronick MN, Grossman PD
JournalClin Chem
PubMed ID6883673
We describe immunoassay techniques that take advantage of the novel properties of phycobiliprotein fluorescent dyes. These dyes, which can be isolated from a wide variety of algae, exhibit extremely high absorptivities, high quantum efficiencies, and excitation and emission bands across the visible spectrum. These stable, hydrophilic proteins can easily be ... More
Separation of phycobiliprotein subunits by reverse-phase high-pressure liquid chromatography.
AuthorsSwanson RV, Glazer AN
JournalAnal Biochem
PubMed ID2221378
Baseline separation of subunits of diverse phycobiliproteins was achieved by a reverse-phase HPLC gradient method with a C4 large-pore column and a solvent system consisting of 0.1% trifluoroacetic acid (TFA) in water and 0.1% TFA in 2:1 (v/v) acetonitrile:isopropanol. The procedure was successfully applied to cyanobacterial allophycocyanin and C-phycocyanins, an ... More
Fluorescent tandem phycobiliprotein conjugates. Emission wavelength shifting by energy transfer.
AuthorsGlazer AN, Stryer L
JournalBiophys J
PubMed ID6414547
A fluorescent tandem phycobiliprotein conjugate with a large Stokes shift was prepared by the covalent attachment of phycoerythrin to allophycocyanin. The efficiency of energy transfer from phycoerythrin to allophycocyanin in this disulfide-linked conjugate was 90%. A distinctive feature of this phycocyanin conjugate is the wide separation between the intense absorption ... More
Photostability studies of phycobiliprotein fluorescent labels.
AuthorsWhite JC, Stryer L
JournalAnal Biochem
PubMed ID3578804
Photostability studies of four fluorescent phycobiliproteins were conducted to identify stable chromophores for biological labeling applications. Phycobiliprotein photodestruction was linear with the applied laser power and depended on the total number of photons absorbed per molecule. Photodestruction quantum yields phi of 1.1 X 10(-5) for R-phycoerythrin, 6.6 X 10(-6) for ... More
Ultrasensitive native fluorescence detection of proteins with miniaturized polyacrylamide gel electrophoresis by laser side-entry excitation.
AuthorsZhang H, Yeung ES
JournalElectrophoresis
PubMed ID16927346
Direct detection of separated proteins inside polyacrylamide gels has many advantages compared to staining methods. Ultrasensitive native fluorescence detection of proteins with miniaturized 1-D and 2-D PAGE was achieved with laser side-entry excitation. The detection limit for R-phycoerythrin protein spots in 1-D SDS-PAGE with 532 nm excitation was as low ... More
Melatonin: a peroxyl radical scavenger more effective than vitamin E.
AuthorsPieri C, Marra M, Moroni F, Recchioni R, Marcheselli F
JournalLife Sci
PubMed ID7934611
We have compared the peroxyl radical scavenger ability to melatonin with that of vitamin E, vitamin C and reduced glutathione (GSH). In the assay system, beta-phycoerythrin (beta-PE) was used as fluorescent indicator protein, 2-2'-azo-bis(2-amidinopropane)dihydrochloride as a peroxyl radical generator and the water soluble vitamin E analogue. Trolox, as reference standard. ... More
Detection of distinct subpopulations of Langerhans cells by flow cytometry and sorting.
AuthorsVaigot P, Czernielewski J, Prunieras M
JournalCytometry
PubMed ID2931262
Flow cytometry was found to be a very appropriate tool for the study of Langerhans cells (LC), which represent a minor cell population (2-3%) of human epidermis, and allowed us to obtain new phenotypic, functional, and cell cycle data on these rare cells. The phenotypic analysis of cell surface antigens ... More
Molecular analysis of the bare lymphocyte syndrome.
AuthorsSullivan KE, Stobo JD, Peterlin BM
JournalJ Clin Invest
PubMed ID3894422
The bare lymphocyte syndrome is a disorder in which class I histocompatibility antigens fail to be expressed normally on the surface of lymphocytes. Utilizing complementary DNA probes for both beta 2-microglobulin and class I genes, the molecular basis for this syndrome was investigated in a family with two siblings exhibiting ... More
Multiparameter analysis of human lymphocyte subpopulations using flow cytometry.
AuthorsLewis DE, Barron KS, Miller GP, Rich RR
JournalSurv Synth Pathol Res
PubMed ID2938244
This review has emphasized several recent advances in our understanding of human lymphocyte biology made possible by mAbs that recognize specific functional subpopulations of lymphocytes, coupled with multiparameter flow-cytometric analysis. In the last 3 years, both the T4+ and T8+ subsets of T cells have been further divided according to ... More
Diphenylamine: an unusual antioxidant.
AuthorsSugihara T, Rao G, Hebbel RP
JournalFree Radic Biol Med
PubMed ID8468023
Diphenylamine (DPA) has been utilized as an antioxidant in studies of lipid peroxidation. Using peroxidizing red blood cell (RBC) membranes, we find that DPA actually promotes lipid hydroperoxide (LOOH) formation and oxygen consumption while markedly inhibiting generation of thiobarbituric acid reactive substances (TBARS). As a consequence, DPA increases the prelytic ... More
The T11 glycoprotein is functionally linked to a calcium channel in precursor and mature T-lineage cells.
AuthorsAlcover A, Weiss MJ, Daley JF, Reinherz EL
JournalProc Natl Acad Sci U S A
PubMed ID2422657
Human T lymphocytes are activated through either the antigen/major histocompatibility complex receptor (T3-Ti) or the T11 sheep erythrocyte-binding protein. Spectrofluorimetry and multiparameter flow cytometric techniques were utilized to examine the relationship of activation to alterations in cytoplasmic free calcium concentration, [Ca2+]i. T3-Ti receptor-triggered elevation in [Ca2+]i was found to be ... More
The optimal application of forward and ninety-degree light scatter in flow cytometry for the gating of mononuclear cells.
AuthorsThompson JM, Gralow JR, Levy R, Miller RA
JournalCytometry
PubMed ID3899551
Peripheral blood mononuclear cells from ten normal donors were labeled with a monoclonal antibody specific for monocytes and analyzed using a fluorescence activated cell sorter (FACS). Forward and 90 degrees light scatter parameters were studied in order to apply optimal computerized gating to identify and exclude monocytes from lymphocyte populations. ... More
Nuclear protein import: specificity for transport across the nuclear pore.
AuthorsWolff B, Willingham MC, Hanover JA
JournalExp Cell Res
PubMed ID3169133
Transport of proteins into the cell nucleus is thought to require specific localization sequences and may be mediated by nuclear pores. Following microinjection into fused cultured cells, nuclear protein import was directly monitored by fluorescence microscopy using B-phycoerythrin (PE; Mr 240,000) coupled to synthetic peptides corresponding to the simian virus ... More
Specific targeting of cytotoxic T cells by anti-T3 linked to anti-target cell antibody.
The specificity of cytotoxic T lymphocytes (Tc) cells is conferred by an antigen-specific receptor, Ti, which in humans is physically associated with an invariant cell-surface glycoprotein, T3. Monoclonal antibodies specific for either T3 and Ti are able to elicit a variety of T-cell responses such as lymphokine production, mitogenesis and ... More
Scanning two-photon fluctuation correlation spectroscopy: particle counting measurements for detection of molecular aggregation.
AuthorsBerland KM, So PT, Chen Y, Mantulin WW, Gratton E
JournalBiophys J
PubMed ID8804624
Scanning fluctuation correlation spectroscopy (FCS) is an experimental technique capable of measuring particle number concentrations by monitoring spontaneous equilibrium fluctuations in the local concentration of a fluorescent species in a small (femtoliter) subvolume of a sample. The method can be used to detect molecular aggregation for dilute, submicromolar samples by ... More
Line-scanning microphotolysis for diffraction-limited measurements of lateral diffusion.
AuthorsWedekind P, Kubitscheck U, Heinrich O, Peters R
JournalBiophys J
PubMed ID8874037
Fluorescence microphotolysis was combined with confocal laser-scanning microscopy to yield a method, herein referred to as line-scanning microphotolysis (LINESCAMP), for the measurement of molecular transport at a lateral resolution of approximately 0.34 microns and a temporal resolution of approximately 0.5 ms. A confocal microscope was operated in the line scan ... More
Human lymphocyte subpopulations identified by using three-color immunofluorescence and flow cytometry analysis: correlation of Leu-2, Leu-3, Leu-7, Leu-8, and Leu-11 cell surface antigen expression.
AuthorsLanier LL, Loken MR
JournalJ Immunol
PubMed ID6361119
Three-color immunofluorescence has been used to determine the co-expression of cell surface antigens on human peripheral blood lymphocytes. Monoclonal antibodies or avidin were coupled to either FITC (green), phycoerythrin (orange), or Texas Red (red) fluorochromes. These three fluorochromes could be independently measured by using a dual laser FACS IV system ... More
Oxidative damage caused by free radicals produced during catecholamine autoxidation: protective effects of O-methylation and melatonin.
AuthorsMiller JW, Selhub J, Joseph JA
JournalFree Radic Biol Med
PubMed ID8818640
Catecholamine autoxidation produces reactive oxygen species that have been implicated in the loss of dopaminergic neurons in the nigrostriatal region of the brain that occurs during normal aging and in Parkinson's disease. In the present study, the potential protective effects of catecholamine O-methylation and of melatonin against catecholamine autoxidation-induced protein ... More
Dendrimer-assisted patch-clamp sizing of nuclear pores.
AuthorsBustamante JO, Michelette ER, Geibel JP, Hanover JA, McDonnell TJ, Dean DA
JournalPflugers Arch
PubMed ID10784359
Macromolecular translocation (MMT) across the nuclear envelope (NE) occurs exclusively through the nuclear pore complex (NPC). Therefore, the diameter of the NPC aqueous/electrolytic channel (NPCC) is important for cellular structure and function. The NPCC diameter was previously determined to be approximately equal to 10 nm with electron microscopy (EM) using ... More
Oxidation of 3-hydroxyanthranilic acid to the phenoxazinone cinnabarinic acid by peroxyl radicals and by compound I of peroxidases or catalase.
AuthorsChristen S, Southwell-Keely PT, Stocker R
JournalBiochemistry
PubMed ID1324727
Since 3-hydroxyanthranilic acid (3HAA), an oxidation product of tryptophan metabolism, is a powerful radical scavenger [Christen, S., Peterhans, E., & Stocker, R. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 2506], its reaction with peroxyl radicals was investigated further. Exposure to aqueous peroxyl radicals generated at constant rate under air from ... More
Determination of co-expression of activation antigens on proliferating CD4+, CD4+ CD8+ and CD8+ lymphocyte subsets by dual parameter flow cytometry.
AuthorsCreemers PC
JournalJ Immunol Methods
PubMed ID2950175
The incidence of activation markers on proliferating CD4+, CD4+ CD8+ and CD8+ lymphocyte subsets was determined in a single laser Epics-C fluorescence-activated cell sorter system, using a series of double staining combinations. Experiments were performed after 3 days of culture with PHA on cell fractions enriched for CD4+ or CD8+ ... More
An optical method for recording the activity of single transporters in membrane patches.
AuthorsTschödrich-Rotter M, Peters R
JournalJ Microsc
PubMed ID9853370
We have previously introduced an optical technique for recording the transport of fluorescent substrates by single membrane transporters. Referred to as optical single-transporter recording (OSTR), the method was restricted to cases in which membrane transporters occurred at extremely small densities, namely at one or a few transporters per cell. Here ... More
Glycosylated hemoglobin and fructosamines: does their determination really reflect the glycemic control in diabetic patients?
AuthorsTesta R, Testa I, Manfrini S, Bonfigli AR, Piantanelli L, Marra M, Pieri C
JournalLife Sci
PubMed ID8684270
The present experiment was designed to determine whether scavenging capacity of serum, in addition to glucose level, influences hemoglobin and serum protein glycosylation in non-insulin dependent diabetic patients. For this purpose forty-seven patients homogeneous for age, disease duration, therapy and glyco-metabolic control were selected. Fasting and post-prandial glycemia and insulinemia ... More
Calcium, ATP and nuclear pore channel gating.
AuthorsBustamante JO, Michelette ER, Geibel JP, Dean DA, Hanover JA, McDonnell TJ
JournalPflugers Arch
PubMed ID10678739
Nuclear envelope (NE) cisternal Ca2+ and cytosolic ATP are required for nuclear-pore-complex-(NPC-) mediated transport of DNAs, RNAs, transcription factors and other large molecules. Isolated cardiomyocyte nuclei, capable of macromolecular transport (MMT), have intrinsic NPC ion channel behavior. The large ion conductance (gamma) activity of the NPC channel (NPCC) is blocked ... More
Two-color flow cytometry and functional analysis of lymphocytes cultured from human renal allografts: identification of a Leu-2+3+ subpopulation.
The phenotype of T lymphocyte subsets present in renal biopsies showing acute cellular allograft rejection in six patients on cyclosporine have been characterized in situ by immunoperoxidase staining, and after expansion in vitro in interleukin 2 (IL-2) by two-color flow cytometry, sorting, and functional analysis. After 8 to 42 days ... More
The ion channel behavior of the nuclear pore complex.
AuthorsBustamante JO, Hanover JA, Liepins A
JournalJ Membr Biol
PubMed ID8568839
Macromolecule-conducting pores have been recently recognized as a distinct class of ion channels. The poor role of macromolecules as electrical charge carriers can be used to detect their movement along electrolyte-filled pores. Because of their negligible contribution to electrical ion currents, translocating macromolecules reduce the net conductivity of the medium ... More
Binding of histamine and histamine analogs to lymphocyte subsets analyzed by flow cytometry.
AuthorsMuirhead K, Bender P, Hanna N, Poste G
JournalJ Immunol
PubMed ID2415600
The binding of histamine, 4-methylhistamine (a histamine type 2 receptor agonist), cimetidine (a histamine type 2 receptor antagonist), and telemethylhistamine (an inactive analog) to human peripheral blood mononuclear cell subsets was investigated by flow cytometry by using conjugates of these ligands coupled to fluorescein-labeled human serum albumin. Our results indicate ... More
Subunit structure and chromophore composition of rhodophytan phycoerythrins. Porphyridium cruentum B-phycoerythrin and b-phycoerythrin.
AuthorsGlazer AN, Hixson CS
JournalJ Biol Chem
PubMed ID833125
A comparative study is presented of the two phycoerythrins of the unicellular red alga Porphyridium cruentum. Native B-phycoerythrin has a molecular weight of 236,000 +/- 18,000 in 0.05 M potassium phosphate at pH 7.0, and an absorption spectrum with maxima at 545 nm (epsilonM = 2.41 X 10(6) M-1 cm-1) ... More
Fluorescent phycobiliprotein conjugates for analyses of cells and molecules.
AuthorsOi VT, Glazer AN, Stryer L
JournalJ Cell Biol
PubMed ID6749865
The synthesis of a novel class of reagents for fluorescence analyses of molecules and cells is reported. These compounds consist of a highly fluorescent phycobiliprotein conjugated to a molecule having biological specificity. Phycoerythrin-immunoglobulin, phycoerythrin-protein A, and phycoerythrin-avidin conjugates were prepared. These conjugates bind specifically to beads containing a covalently attached ... More
Detection of triple antibody-binding lymphocytes in standard single laser flow cytometry using colloidal gold, fluorescein and phycoerythrin as labels.
AuthorsFestin R, Björklund B, Tötterman TH
JournalJ Immunol Methods
PubMed ID2956330
Goat anti-mouse antibodies conjugated with colloidal 40 nm gold particles (G40) were used as secondary layers to stain human T lymphocytes in an attempt to extend the polarized light epi-illumination microscopic technique to flow cytometric multiparameter analysis. G40-labelled T cells were further stained with phycoerythrin (PE, red)- and fluorescein (FITC, ... More
Nucleocytoplasmic protein traffic in single mammalian cells studied by fluorescence microphotolysis.
AuthorsSchulz B, Peters R
JournalBiochim Biophys Acta
PubMed ID3651509
Fluorescence microphotolysis was employed to measure in single living cells the kinetics of nucleocytoplasmic transport and the coefficients of intracellular diffusional mobility for the nuclear non-chromosomal protein nucleoplasmin. Nucleoplasmin was isolated from Xenopus ovary and labeled fluorescently. By injection into Xenopus oocytes it was ascertained that fluorescent labeling did not ... More
Peroxyl radical scavenging activity of Ginkgo biloba extract EGb 761.
AuthorsMaitra I, Marcocci L, Droy-Lefaix MT, Packer L
JournalBiochem Pharmacol
PubMed ID7786306
Antioxidant mechanisms have been proposed to underlie the beneficial pharmacological effects of EGb 761, an extract from Ginkgo biloba leaves used for treating peripheral vascular diseases and cerebrovascular insufficiency in the elderly. In vitro evidence has been reported that EGb 761 scavenges various reactive oxygen species, i.e. nitric oxide, and ... More
Oxygen-radical absorbance capacity assay for antioxidants.
AuthorsCao G, Alessio HM, Cutler RG
JournalFree Radic Biol Med
PubMed ID8458588
A relatively simple but sensitive and reliable method of quantitating the oxygen-radical absorbing capacity (ORAC) of antioxidants in serum using a few microliter is described. In this assay system, beta-phycoerythrin (beta-PE) is used as an indicator protein, 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) as a peroxyl radical generator, and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox, a ... More
Autoxidative and antioxidative potential of simple carbohydrates.
AuthorsWehmeier KR, Mooradian AD
JournalFree Radic Biol Med
PubMed ID7959169
To determine the relative ranking of autoxidative potential of various sugars the fluorescence of phycoerythrin was monitored in the presence of various concentrations of sugars incubated at 37 degrees C with or without CuSO4. The antioxidative properties of these sugars was assayed in the presence of a peroxy radical generator ... More
In vitro study on antioxidant potential of various drugs used in the perioperative period.
AuthorsKang MY, Tsuchiya M, Packer L, Manabe M
JournalActa Anaesthesiol Scand
PubMed ID9527743
BACKGROUND: Since surgical trauma not only intensifies the oxidative stress by generating reactive oxygen species (ROS), but also weakens the biological defense system against ROS attack, the antioxidant activity of drugs used during the perioperative period, which possibly normalizes the impaired redox state in the patient, is of fundamental importance ... More
Improved flow cytometric analysis of leukocyte subsets: simultaneous identification of five cell subsets using two-color immunofluorescence.
AuthorsHoran PK, Slezak SE, Poste G
JournalProc Natl Acad Sci U S A
PubMed ID3534887
Flow cytometric analysis of human peripheral blood leukocytes has typically been achieved by staining multiple aliquots of the same sample with fluorescent reagents specific for cell subsets of interest. Spectrally discrete fluorochrome tags have been developed for applications in which identification of multiple subsets (e.g., T and B cells) or ... More
In vitro transport of a fluorescent nuclear protein and exclusion of non-nuclear proteins.
AuthorsNewmeyer DD, Finlay DR, Forbes DJ
JournalJ Cell Biol
PubMed ID3097026
An in vitro system was developed that provides a quick microscopic assay for nuclear transport. The assay uses an extract of Xenopus eggs, normal or synthetic nuclei, and a fluorescently labeled nuclear protein, nucleoplasmin. This in vitro system accurately mimics in vivo nuclear transport, both in exclusivity and in the ... More