Enzymatic isolation and characterization of single vascular smooth muscle cells from cremasteric arterioles.
AuthorsJackson WF,Huebner JM,Rusch NJ
JournalMicrocirculation (New York, N.Y. : 1994)
PubMed ID9110282
OBJECTIVE: The goal of the present study was to develop a method to isolate viable arteriolar muscle cells from single cremasteric arterioles, which retain the contractile and electrophysiological phenotype of the donor microvessels. METHODS: Arterioles were hand-dissected from rat and hamster cremaster muscles and dissociated by incubation in papain and ... More
Enzymatic isolation and characterization of single vascular smooth muscle cells from cremasteric arterioles.
AuthorsJackson WF,Huebner JM,Rusch NJ
JournalMicrocirculation (New York, N.Y. : 1994)
PubMed ID8930888
OBJECTIVE: The goal of the present study was to develop a method to isolate enzymatically viable arteriolar muscle cells from single cremasteric arterioles, which retain the contractile and electrophysiological phenotype of the donor microvessels. METHODS: Arterioles were hand-dissected from rat and hamster cremaster muscles and dissociated by incubation in papain ... More
A stable double-stranded DNA-ethidium homodimer complex: application to picogram fluorescence detection of DNA in agarose gels.
AuthorsGlazer AN, Peck K, Mathies RA
JournalProc Natl Acad Sci U S A
PubMed ID2339125
The complex between double-stranded DNA and ethidium homodimer (5,5'-diazadecamethylene)bis(3,8-diamino-6-phenylphenanthridini um) cation, formed at a ratio of 1 homodimer per 4 or 5 base pairs, is stable in agarose gels under the usual conditions for electrophoresis. This unusual stability allows formation of the complex before electrophoresis and then separation and detection ... More
Successful storage of peripheral nerve before transplantation using green tea polyphenol: an experimental study in rats.
Green tea polyphenol is known to act as a buffer, reducing biological responses to oxidative stress. Several effects of polyphenol have been reported, such as protection of tissue from ischemia, antineoplasmic and anti-inflammatory effects, and suppression of arteriosclerosis. In this study, we investigated whether peripheral nerve segments could be kept ... More
Indirubin enhances tumor necrosis factor-induced apoptosis through modulation of nuclear factor-kappa B signaling pathway.
AuthorsSethi G, Ahn KS, Sandur SK, Lin X, Chaturvedi MM, Aggarwal BB
JournalJ Biol Chem
PubMed ID16785236
'Although indirubin is known to exhibit anti-cancer and anti-inflammatory activities, very little is known about its mechanism of action. In this study, we investigated whether indirubin mediates its effects through interference with the NF-kappaB pathway. As examined by the DNA binding of NF-kappaB, we found that indirubin suppressed tumor necrosis ... More
The apoptotic regulatory protein ARC (apoptosis repressor with caspase recruitment domain) prevents oxidant stress-mediated cell death by preserving mitochondrial function.
Authors Neuss M; Monticone R; Lundberg M S; Chesley A T; Fleck E; Crow M T;
JournalJ Biol Chem
PubMed ID11438535
'ARC is an apoptotic regulatory protein expressed almost exclusively in myogenic cells. It contains a caspase recruitment domain (CARD) through which it has been shown to block the activation of some initiator caspases. Because ARC also blocks caspase-independent events associated with apoptosis, such as hypoxia-induced cytochrome c release, we examined ... More
Redistribution and dysfunction of integrins in cultured renal epithelial cells exposed to oxidative stress.
AuthorsGailit J, Colflesh D, Rabiner I, Simone J, Goligorsky MS
JournalAm J Physiol
PubMed ID8430825
'Tubular obstruction by detached renal tubular epithelial cells is a major cause of oliguria in acute renal failure. Viable renal tubular cells can be recovered from urine of patients with acute tubular necrosis, suggesting a possible defect in cell adhesion to the basement membrane. To study this process of epithelial ... More
Simultaneous expression of c-Jun and p53 in retinal ganglion cells of adult rat retinal slice cultures.
AuthorsUmihira J, Lindsey JD, Weinreb RN
JournalCurr Eye Res
PubMed ID12187487
'PURPOSE: To determine whether the apoptosis-associated transcription factor c-Jun and the regulator protein p53 are expressed together during retinal ganglion cell (RGC) death in slice cultures of adult rat retina, and whether c-Jun expression or p53 expression is altered by glutamate. METHODS: Newborn rat RGCs were retrogradely labeled by Di-I ... More
In vitro characterization of a bone marrow stem cell-seeded collagen gel composite for soft tissue grafts: effects of fiber number and serum concentration.
AuthorsLewus KE, Nauman EA
JournalTissue Eng
PubMed ID16144437
'Cell-seeded collagen hydrogels have been used in the engineering of many tissue types, from skin and vasculature to spinal cord. One of the primary limitations of collagen-based hydrogels for use in tissue-engineered grafts is that cells seeded within the gel cause it to contract as much as 70%. By forming ... More
Ethidium monoazide for DNA-based differentiation of viable and dead bacteria by 5'-nuclease PCR.
AuthorsNogva HK, Drømtorp SM, Nissen H, Rudi K
JournalBiotechniques
PubMed ID12703305
'PCR techniques have significantly improved the detection and identification of bacterial pathogens. Even so, the lack of differentiation between DNA from viable and dead cells is one of the major challenges for diagnostic DNA-based methods. Certain nucleic acid-binding dyes can selectively enter dead bacteria and subsequently be covalently linked to ... More
Difference in volume of X- and Y-chromosome-bearing bovine sperm heads matches difference in DNA content.
Authorsvan Munster EB, Stap J, Hoebe RA, te Meerman GJ, Aten JA
JournalCytometry
PubMed ID10554167
'BACKGROUND: To investigate the possibilities of sperm head volume as a sorting criterion for gender preselection, we determined the magnitude of the difference in volume of X- and Y-chromosome-bearing bull sperm heads. MATERIALS AND METHODS: Bovine sperm heads were sorted on the basis of their DNA content in X- and ... 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
Controlling hydrogelation kinetics by peptide design for three-dimensional encapsulation and injectable delivery of cells.
AuthorsHaines-Butterick L, Rajagopal K, Branco M, Salick D, Rughani R, Pilarz M, Lamm MS, Pochan DJ, Schneider JP
JournalProc Natl Acad Sci U S A
PubMed ID17470802
'A peptide-based hydrogelation strategy has been developed that allows homogenous encapsulation and subsequent delivery of C3H10t1/2 mesenchymal stem cells. Structure-based peptide design afforded MAX8, a 20-residue peptide that folds and self-assembles in response to DMEM resulting in mechanically rigid hydrogels. The folding and self-assembly kinetics of MAX8 have been tuned ... More
Confocal in vivo microscopy and confocal laser-scanning fluorescence microscopy in keratoconus.
AuthorsSomodi S, Hahnel C, Slowik C, Richter A, Weiss DG, Guthoff R
JournalGer J Ophthalmol
PubMed ID9479549
'The purpose of this study was the determination of morphological changes in the corneal epithelium and the keratocyte network in keratoconus. In all, 33 eyes of 19 patients were examined in vivo using the confocal slit-scanning microscope Microphthal. After penetrating keratoplasty, recipients'' trephanates were stained with the Live/Dead kit and ... More
Thermodynamic investigation of the association of ethidium, propidium and bis-ethidium to DNA hairpins.
AuthorsRentzeperis D, Medero M, Marky LA
JournalBioorg Med Chem
PubMed ID7582953
'We have used a combination of calorimetric and spectroscopic techniques to investigate the association of the bis-intercalator ethidium homodimer (bis-ethidium) to short DNA hairpins with sequences: d(GCGCT5GCGC) and d(CGCGT5CGCG). The helix-coil transition of each hairpin, investigated by UV and calorimetric melting protocol, takes place in monomolecular two-state transitions with characteristic ... More
Cadmium alters the localization of N-cadherin, E-cadherin, and beta-catenin in the proximal tubule epithelium.
AuthorsProzialeck WC, Lamar PC, Lynch SM
JournalToxicol Appl Pharmacol
PubMed ID12791303
'Recent studies on proximal tubule-derived cells in culture have shown that Cd has relatively specific damaging effects on the cadherin-dependent junctions between the cells. The objective of the present study was to determine whether Cd can affect cadherin-dependent junctions in the proximal tubule epithelium in vivo. Male Sprague-Dawley rats received ... More
Heterodimeric DNA-binding dyes designed for energy transfer: stability and applications of the DNA complexes.
AuthorsBenson SC, Mathies RA, Glazer AN
JournalNucleic Acids Res
PubMed ID8284220
'Spectroscopic studies of the complexes of double-stranded (ds) DNA with the polymethylene-amine linked heterodimers thiazole orange-thiazole blue, thiazole orange-ethidium, and fluorescein-ethidium, in each case show efficient energy transfer from donor to acceptor chromophores (Benson, S.C., Singh, P. and Glazer, A.N. (1993) accompanying manuscript). A quantitative assay of the stability of ... More
Protection from apoptosis in human neutrophils is determined by the surface of adhesion.
AuthorsGinis I, Faller DV
JournalAm J Physiol
PubMed ID9038836
'Recent work suggests that various neutrophil agonists affect the rate of apoptosis in these cells. On the basis of these observations, we hypothesized that signals triggered in neutrophils via their adhesion receptors might also modify their life span. This hypothesis has been tested using human neutrophils adherent to tissue culture ... More
Stable fluorescent complexes of double-stranded DNA with bis-intercalating asymmetric cyanine dyes: properties and applications.
AuthorsRye HS, Yue S, Wemmer DE, Quesada MA, Haugland RP, Mathies RA, Glazer AN
JournalNucleic Acids Res
PubMed ID1614866
'The synthesis, proof of structure, and the absorption and fluorescence properties of two new unsymmetrical cyanine dyes, thiazole orange dimer (TOTO; 1,1''-(4,4,7,7-tetramethyl-4,7- diazaundecamethylene)-bis-4-[3-methyl-2,3-dihydro-(benzo-1,3-thiaz ole)-2- methylidene]-quinolinium tetraiodide) and oxazole yellow dimer (YOYO; an analogue of TOTO with a benzo-1,3-oxazole in place of the benzo-1,3-thiazole) are reported. TOTO and YOYO are virtually ... More
Effects of cadmium on E-cadherin and VE-cadherin in mouse lung.
AuthorsPearson CA, Lamar PC, Prozialeck WC
JournalLife Sci
PubMed ID12570930
'Exposure to Cd(2+) via inhalation or intratracheal instillation results in pulmonary edema, which is followed by the influx of leukocytes, the proliferation of type II pneumocytes and eventual scarring and fibrotic changes. While the general toxic effects of Cd(2+) in the lung have been well characterized, the specific molecular mechanisms ... More
Intercalative interactions of ethidium dyes with triplex structures.
AuthorsTuite E, Nordén B
JournalBioorg Med Chem
PubMed ID7582948
'The binding of phenanthridine dyes to triplex poly(dT)*poly(dA).poly(dT) and its precursor duplex poly(dA).poly(dT) is characterized using linear dichroism and circular dichroism spectroscopy, and thermal denaturation. The two monomeric dyes ethidium bromide and propidium iodide are shown to behave similarly to each other in intercalating into and stabilizing both the duplex ... More
Spectrofluorometric assay for hybridization of oligodeoxynucleotides using ethidium dimer.
AuthorsTombler ER, Deutsch DG
JournalBiotechniques
PubMed ID8292339
'A spectrofluorometric assay to detect hybridization of oligodeoxynucleotides is described. Ethidium dimer, which yields significant fluorescence enhancement when intercalated between adjacent base pairs, was employed as the probe. Maximum enhancement of fluorescence occurred at equimolar ratios of p(dA)20.p(dT)20, while negligible fluorescence occurred for either of these chains when not in ... More
Cytotoxic effect of beta-amyloid on a human differentiated neuron is not mediated by cytoplasmic Ca2+ accumulation.
AuthorsGao ZY, Collins HW, Matschinsky FM, Lee VM, Wolf BA
JournalJ Neurochem
PubMed ID9523555
'The effects of synthetic beta-amyloid (A beta1-42) on cell viability and cellular Ca2+ homeostasis have been studied in the human neuron-like NT2N cell, which differentiates from a teratocarcinoma cell line, NTera2/C1.D1, by retinoic acid treatment. NT2N viability was measured using morphological criteria and fluorescent live/dead staining and quantified using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium ... More
High-sensitivity capillary electrophoresis of double-stranded DNA fragments using monomeric and dimeric fluorescent intercalating dyes.
AuthorsZhu H, Clark SM, Benson SC, Rye HS, Glazer AN, Mathies RA
JournalAnal Chem
PubMed ID8067520
'Fluorescence-detected capillary electrophoresis separations of phi X174/HaeIII DNA restriction fragments have been performed using monomeric and dimeric intercalating dyes. Replaceable hydroxyethyl cellulose solutions were used as the separation medium. Confocal fluorescence detection was performed following 488-nm laser excitation. The limits of DNA detection for on-column staining with monomeric dyes (ethidium ... More
In vitro pharmacology of cryptophycin 52 (LY355703) in human tumor cell lines.
AuthorsWagner MM, Paul DC, Shih C, Jordan MA, Wilson L, Williams DC
JournalCancer Chemother Pharmacol
PubMed ID9923816
'PURPOSE: Cryptophycin 52 (LY355703) is a new member of the cryptophycin family of antitumor agents that is currently undergoing clinical evaluation for cancer chemotherapy. The mechanism of action of the cryptophycin class of compounds is associated with an action on microtubules. This report details the pharmacological profile of this new ... More
Green-light transilluminator for the detection without photodamage of proteins and DNA labeled with different fluorescent dyes.
AuthorsAlba FJ, Bermúdez A, Daban JR
JournalElectrophoresis
PubMed ID11258745
'The excitation spectra of Nile red and SYPRO red, two currently used dyes for the fluorescent staining of protein bands in sodium dodecyl sulfate (SDS)-polyacrylamide gels, show an excitation peak in the UV region and another in the visible region (maximum at about 550 nm). Ethidium bromide and other intercalating ... More
Co-extrusion of biocompatible polymers for scaffolds with co-continuous morphology.
AuthorsWashburn NR, Simon CG, Tona A, Elgendy HM, Karim A, Amis EJ
JournalJ Biomed Mater Res
PubMed ID11835155
'A methodology for the preparation of porous scaffolds for tissue engineering using co-extrusion is presented. Poly(epsilon-caprolactone) is blended with poly(ethylene oxide) in a twinscrew extruder to form a two-phase material with micron-sized domains. Selective dissolution of the poly(ethylene oxide) with water results in a porous material. A range of blend ... More
Constitutive expression of the machinery for programmed cell death.
AuthorsWeil M, Jacobson MD, Coles HS, Davies TJ, Gardner RL, Raff KD, Raff MC
JournalJ Cell Biol
PubMed ID8655578
'In the presence of cycloheximide (CHX) to inhibit protein synthesis, a high concentration of staurosporine (STS) induces almost all cells in explant cultures of 8/8 types of newborn mouse organs and 3/3 types of adult mouse organs to die with the characteristic features of apoptosis. Eggs and blastomeres also die ... More
Morphine does not affect astrocyte survival in developing primary mixed-glial cultures.
AuthorsGurwell JA, Hauser KF
JournalBrain Res Dev Brain Res
PubMed ID8149599
'In mixed-glial cultures, high concentrations of morphine (1 microM) have previously been shown to completely inhibit any increase in glial numbers, although DNA synthesis continues in flat, polyhedral astrocytes (type 1 astrocytes). This suggests that high concentrations of morphine are toxic to glia. Morphine toxicity was assessed in mixed-glial cultures ... More
Measurement of tumor necrosis factor activity by flow cytometry.
AuthorsLévesque A, Paquet A, Pagé M
JournalCytometry
PubMed ID7664628
'Tumor necrosis factor-alpha (TNF-alpha) is a monokine of 17 kDa produced by activated macrophages and various cells involved in the immune system. We propose a new method for the measurement of TNF activity using flow cytometry. After an incubation with TNF, L929 cells were harvested and treated with a calcein-AM ... More
Interaction of dimeric intercalating dyes with single-stranded DNA.
AuthorsRye HS, Glazer AN
JournalNucleic Acids Res
PubMed ID7739900
'The unsymmetrical cyanine dye thiazole orange homodimer (TOTO) binds to single-stranded DNA (ssDNA, M13mp18 ssDNA) to form a fluorescent complex that is stable under the standard conditions of electrophoresis. The stability of this complex is indistinguishable from that of the corresponding complex of TOTO with double-stranded DNA (dsDNA). To examine ... More
Hypoxic preconditioning protects cultured neurons against hypoxic stress via TNF-alpha and ceramide.
AuthorsLiu J, Ginis I, Spatz M, Hallenbeck JM
JournalAm J Physiol Cell Physiol
PubMed ID10644522
'Brief "preconditioning" ischemia induces ischemic tolerance (IT) and protects the animal brain from subsequent otherwise lethal ischemia. Identification of the signaling steps most proximal to the development of the IT will allow induction of the resistance to ischemia shortly after the onset of stroke. Animal studies demonstrate a key role ... More
Mechanism of oxidative stress from low levels of carbon monoxide.
AuthorsThom SR, Ischiropoulos H
JournalRes Rep Health Eff Inst
PubMed ID9476263
'The purpose of this study was to determine whether platelets and vascular endothelial cells would liberate nitric oxide free radical (NO)* and NO-derived oxidant species after exposure to carbon monoxide (CO) at concentrations up to 100 parts per million (ppm). We hypothesized that exposure to environmentally relevant concentrations of CO ... More
Nucleoplasmic Ca(2+)loading is regulated by mobilization of perinuclear Ca(2+).
AuthorsAbrenica B, Gilchrist JS
JournalCell Calcium
PubMed ID10970769
'Regulation of nucleoplasmic calcium (Ca(2+)) concentration may occur by the mobilization of perinuclear luminal Ca(2+)pools involving specific Ca(2+)pumps and channels of both inner and outer perinuclear membranes. To determine the role of perinuclear luminal Ca(2+), we examined freshly cultured 10 day-old embryonic chick ventricular cardiomyocytes. We obtained evidence suggesting the ... More
Apoptosis-inducing factor is involved in the regulation of caspase-independent neuronal cell death.
'Caspase-independent death mechanisms have been shown to execute apoptosis in many types of neuronal injury. P53 has been identified as a key regulator of neuronal cell death after acute injury such as DNA damage, ischemia, and excitotoxicity. Here, we demonstrate that p53 can induce neuronal cell death via a caspase-mediated ... More
Vital staining of the hearing organ: visualization of cellular structure with confocal microscopy.
AuthorsFlock A, Scarfone E, Ulfendahl M
JournalNeuroscience
PubMed ID9466411
'Cells inside the intact organ of Corti were labelled with fluorescent probes reflecting various aspects of structure and function. The dyes were introduced into the perilymphatic space by perfusion of the scala tympani of the temporal bone from the guinea-pig maintained in isolation. The dyes were able to diffuse through ... More
Astrocyte Na+ channels are required for maintenance of Na+/K(+)-ATPase activity.
AuthorsSontheimer H, Fernandez-Marques E, Ullrich N, Pappas CA, Waxman SG
JournalJ Neurosci
PubMed ID8182422
'Astrocytes in vitro and in situ have been shown to express voltage-activated ion channels previously thought to be restricted to excitable cells, including voltage-activated Na+, Ca2+, and K+ channels. However, unlike neurons, astrocytes do not generate action potentials, and the functional role of voltage-activated channels in astrocytes has been an ... More
A fluorescence confocal assay to assess neuronal viability in brain slices.
AuthorsMonette R, Small DL, Mealing G, Morley P
JournalBrain Res Brain Res Protoc
PubMed ID9473610
'Hippocampal slice models are used to study the mechanisms of ischemia-induced neurotoxicity and to assess the neuroprotective potential of novel therapeutic agents. A number of morphological and functional endpoints are available to assess neuronal viability. The slice model also allows the study of selectively vulnerable neuronal populations within the same ... More
Fluorescence microscopy and three-dimensional imaging of the porcine corneal keratocyte network.
'BACKGROUND: Little is known about the spatial arrangement and the corresponding morphometric data describing the living keratocyte network. For determination of alterations in corneal diseases it is crucial to know the morphology of the keratocyte network in the healthy state. Porcine cornea was used as a model tissue because it ... More
DNA bifunctional intercalators. I. Synthesis and conformational properties of an ethidium homodimer and of an acridine ethidium heterodimer.
'An ethidium homodimer and an acridine ethidium heterodimer have been synthesized. The ethidium and the acridine chromophore were introduced in such bifunctional intercalators in order to allow the fluorometric study of the interaction of such molecules with DNA, which is reported in the companion paper (Gaugain, B., Barbet, J., Capelle, ... More
Fluoroprobe quantification of viable and non-viable cells in human coronary and internal thoracic arteries sampled at autopsy.
AuthorsMerrilees MJ, Beaumont BW, Scott LJ
JournalJ Vasc Res
PubMed ID8562809
'Viable and non-viable cells in coronary and internal thoracic arteries, collected at autopsy 7-24 h post-mortem from individuals 15-81 years of age, were detected using the fixable fluoroprobes 5-chloromethylfluorescein diacetate (green) and ethidium homodimer-1 (orange/red). Viability status of individual endothelial and smooth muscle cells was confirmed by simultaneous autoradiographic detection ... More
Tissue engineering of vascularized cardiac muscle from human embryonic stem cells.
AuthorsCaspi O, Lesman A, Basevitch Y, Gepstein A, Arbel G, Habib IH, Gepstein L, Levenberg S
JournalCirc Res
PubMed ID17218605
'Transplantation of a tissue-engineered heart muscle represents a novel experimental therapeutic paradigm for myocardial diseases. However, this strategy has been hampered by the lack of sources for human cardiomyocytes and by the scarce vasculature in the ischemic area limiting the engraftment and survival of the transplanted muscle. Beyond the necessity ... More
Equilibrium and kinetic measurements of muscarinic receptor antagonism on living cells using bead injection spectroscopy.
AuthorsHodder PS, Beeson C, Ruzicka J
JournalAnal Chem
PubMed ID10939374
'Bead injection spectroscopy (BIS) techniques are introduced for automated measurement of pharmacological antagonism by functional assay. Chinese hamster ovary cells that express the rat type 1 muscarinic receptor are cultured on microbeads and used as a renewable biological target for muscarinic receptor antagonist ligands. A flow injection instrument is used ... More
Induction of permeability changes and death of vertebrate cells is modulated by the virulence of Entamoeba spp. isolates.
AuthorsDvorak JA, Kobayashi S, Nozaki T, Takeuchi T, Matsubara C
JournalParasitol Int
PubMed ID12798929
'Although Entamoeba histolytica is capable of inducing an apoptotic response in vertebrate cells in vitro (Cell. Microbiol. 2 (2000) 617), it is not known whether vertebrate cell death requires direct amoeba-vertebrate cell contact or simply the presence of amoebae in the area of the vertebrate cells. In addition, Entamoeba spp. ... More
Inhibition of neuronal apoptosis by a metalloporphyrin superoxide dismutase mimic.
AuthorsPatel M
JournalJ Neurochem
PubMed ID9721731
'The objective of this study was to determine whether free radicals play a pathogenic role in neuronal apoptosis. The ability of Mn(III) tetrakis(benzoic acid) porphyrin (MnTBAP), a superoxide dismutase mimic, to inhibit staurosporine-induced neuronal apoptosis was tested in mixed cerebrocortical cultures. Staurosporine produced concentration-dependent cell death that was markedly inhibited ... More
Apoptosis and accidental cell death in cultured human keratinocytes after thermal injury.
'The respective roles of apoptosis and accidental cell death after thermal injury were evaluated in normal human epidermal keratinocytes. By coupling the LIVE/DEAD fluorescence viability assay with the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) method and ultrastructural morphology, these two processes could be distinguished. Cells were grown on glass ... 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
Separation of nucleic acids by capillary electrophoresis in cellulose solutions with mono- and bis-intercalating dyes.
AuthorsKim Y, Morris MD
JournalAnal Chem
PubMed ID8160960
'The effects of bis- and mono-intercalating dyes on the capillary electrophoretic separation of double-stranded DNA have been investigated in buffers containing (hydroxypropylmethyl)-cellulose. Broad bands and incomplete separations of phi X 174 DNA HaeIII digest were obtained with the bis-intercalators ethidium homodimer 1 (EthD-1) and ethidium homodimer 2 (EthD-2), under a ... More
Death of oligodendrocytes mediated by the interaction of nerve growth factor with its receptor p75.
AuthorsCasaccia-Bonnefil P, Carter BD, Dobrowsky RT, Chao MV
JournalNature
PubMed ID8878481
'Members of the nerve growth factor (NGF) family promote the survival of neurons during development. NGF specifically activates the receptor trkA, initiating a signal transduction cascade which ultimately blocks cell death. Here we show that NGF can have the opposite effect, inducing the death of mature oligodendrocytes cultured from postnatal ... More
Uptake, degradation, and release of fibrillar and soluble forms of Alzheimer's amyloid beta-peptide by microglial cells.
AuthorsChung H, Brazil MI, Soe TT, Maxfield FR
JournalJ Biol Chem
PubMed ID10542270
'Microglia are phagocytic cells that are the main inflammatory response cells of the central nervous system. In Alzheimer''s disease brain, activated microglia are concentrated in regions of compact amyloid deposits that contain the 39-43-amino acid Abeta peptide. We examined the uptake, degradation, and release of small aggregates of fibrillar Abeta ... More
Stable dye-DNA intercalation complexes as reagents for high-sensitivity fluorescence detection.
AuthorsGlazer AN, Rye HS
JournalNature
PubMed ID1436062
'Fluorescent intercalation complexes of certain polycationic ligands with double-stranded DNA provide a new class of multichromophore labels for fluorescence assays.'
Evaluation of quantum dot cytotoxicity based on intracellular uptake.
AuthorsChang E, Thekkek N, Yu WW, Colvin VL, Drezek R
JournalSmall
PubMed ID17192996
Advanced image analysis systems in cell, molecular and neurobiology applications.
AuthorsRamm P
JournalJ Neurosci Methods
PubMed ID7869747
'Many of today''s image acquisition devices (scanners and cameras) yield high-resolution (1280 x 1024 pixels is typical) and/or high-precision (> 8 bits) image data. When coupled to a powerful image analyzer these image acquisition devices offer practical advantages in many bioscience applications. The benefits of high resolution include better contrast ... More
A novel, simple organotypic culture method to study the organ of Corti from the neonatal gerbil.
AuthorsLiu TC, He DZ, Lin X
JournalORL J Otorhinolaryngol Relat Spec
PubMed ID9411319
'An original, simple organotypic culture method was developed to grow the organ of Corti from the neonatal gerbil on the bottom of a Petri dish. In comparison with the commonly used Maximov slide assembly method, this method is easier, less time-consuming, and more economic. Our results in this study using ... More
Cell death and proliferation in acute slices and organotypic cultures of mammalian CNS.
AuthorsLossi L, Alasia S, Salio C, Merighi A,
JournalProg Neurobiol
PubMed ID19552996
'Analysis of the interplay between cell proliferation and death has been greatly advantaged by the development of CNS slice preparations. In slices, interactions between neurons and neurons and the glial cells are fundamentally preserved in a fashion close to the in vivo situation. In parallel, these preparations offer the possibility ... More
Differential trafficking of live and dead Mycobacterium marinum organisms in macrophages.
AuthorsBarker LP, George KM, Falkow S, Small PL
JournalInfect Immun
PubMed ID9119492
'We characterized the Mycobacterium marinum phagosome by using a variety of endocytic markers to follow the path of the bacteria through a mouse macrophage cell line. Using a laser confocal microscope, we found that the majority of viable M. marinum cells were in nonacidic vacuoles that did not colocalize with ... More
The keratocyte network of human cornea: a three-dimensional study using confocal laser scanning fluorescence microscopy.
AuthorsHahnel C, Somodi S, Weiss DG, Guthoff RF
JournalCornea
PubMed ID10746451
'PURPOSE: Keratocytes of the living human cornea were examined to compare quantitatively spatial arrangement and cell volume of the stromal layers. This knowledge is required for further studies toward a quantitative understanding of cellular alterations in corneal pathology. METHODS: Three human corneas were stained with calcein AM and ethidium homodimer ... More
Reduction in the E2k subunit of the alpha-ketoglutarate dehydrogenase complex has effects independent of complex activity.
AuthorsShi Q, Chen HL, Xu H, Gibson GE
JournalJ Biol Chem
PubMed ID15649899
'The activity of the alpha-ketoglutarate dehydrogenase complex (KGDHC) declines in brains of patients with several neurodegenerative diseases. KGDHC consists of multiple copies of E1k, E2k, and E3. E1k and E2k are unique to KGDHC and may have functions independent of the complex. The present study tested the consequences of different ... More
DNA Bifunctional intercalators. 2. Fluorescence properties and DNA binding interaction of an ethidium homodimer and an acridine ethidium heterodimer.
AuthorsGaugain B, Barbet J, Capelle N, Roques BP, Le Pecq JB
JournalBiochemistry
PubMed ID569495
'An ethidium homodimer and acridine ethidium heterodimer have been synthesized (Gaugain, B., Barbet, J., Oberlin, R., Roques, B. P., & Le Pecq, J. B. (1978) Biochemistry 17 (preceding paper in this issue)). The binding of these molecules to DNA has been studied. We show that these dimers intercalate only one ... More
[Visualization of keratocytes in the human cornea with fluorescence microscopy]
AuthorsSomodi S, Guthoff R
JournalOphthalmologe
PubMed ID7549328
'There is little information about the state of human keratocytes in longterm cornea storage and the influence of these cells on the survival of the corneal transplant after keratoplasty. Up to now stromal keratocytes have not been taken into consideration in regard to the culture conditions in cornea banking. Therefore, ... More
Viability of human corneal endothelium following Optisol-GS storage.
AuthorsMeans TL, Geroski DH, Hadley A, Lynn MJ, Edelhauser HF
JournalArch Ophthalmol
PubMed ID7786224
'OBJECTIVES: To evaluate endothelial viability of human corneas stored in glass vials and in viewing chambers (Alcon) for extended periods, and to compare endothelial viability of Optisol-GS-stored corneas with corneas excised from moist chamber-stored globes. METHODS: Endothelial viability was assessed using two staining techniques. Endothelium from stored corneas was stained ... More
Anti-phospholipid autoantibodies bind to apoptotic, but not viable, thymocytes in a beta 2-glycoprotein I-dependent manner.
'Anti-phospholipid autoantibodies (aPL) are associated with a clinical syndrome of hypercoagulability, thrombocytopenia, and fetal loss. Several groups have shown that the in vitro target of many aPL is not a pure phospholipid Ag, but is either a complex between anionic phospholipid and the plasma protein beta2-glycoprotein I (beta 2GPI) or ... More
A flow cytometric technique using peanut agglutinin for evaluating acrosomal loss from human spermatozoa.
AuthorsCooper TG, Yeung CH
JournalJ Androl
PubMed ID9796613
'Flow cytometric methods for the quantification of acrosome-reacted ejaculated human spermatozoa are described in which fluorescence-labeled peanut agglutinin (Arachis hypogaea) binding to the outer acrosomal membrane is used after incubation with ethidium homodimer (EHD) as vital dye and membrane permeation using methanol. Fluorescein-labeled fucoidan (F-fucoidin) was shown to bind to ... More
Characteristics of different nucleic acid staining dyes for DNA fragment sizing by flow cytometry.
AuthorsYan X, Grace WK, Yoshida TM, Habbersett RC, Velappan N, Jett JH, Keller RA, Marrone BL
JournalAnal Chem
PubMed ID10624155
'An efficient and reliable double-stranded DNA (dsDNA) staining protocol for DNA fragment sizing by flow cytometry is presented. The protocol employs 0.8 microM of PicoGreen to label a wide range of DNA concentrations (0.5 ng/mL to 10,000 ng/mL) without regard to the solution dye/bp ratios and without initial quantification of ... More
Measurement of NAD(P)H oxidase-derived superoxide with the luminol analogue L-012.
AuthorsDaiber A, August M, Baldus S, Wendt M, Oelze M, Sydow K, Kleschyov AL, Munzel T
JournalFree Radic Biol Med
PubMed ID14732294
'In the present study we sought to determine the ability of the chemiluminescence dye 8-amino-5-chloro-7-phenylpyridol[3,4-d]pyridazine-1,4-(2H,3H)dione sodium salt (L-012) to detect superoxide in different biological systems. In human whole blood or isolated leukocytes, the sensitivity of the luminol analogue L-012 to detect superoxide was higher as compared with luminol, lucigenin, coelenterazine, ... More
Membrane permeabilization induced by discodermin A, a novel marine bioactive peptide.
AuthorsSato K, Horibe K, Amano K, Mitusi-Saito M, Hori M, Matsunaga S, Fusetani N, Ozaki H, Karaki H
JournalToxicon
PubMed ID10978743
'The effects of discodermin A (DC-A), a novel marine bioactive peptide extracted from sea sponge Discodermia kiiensis, on the vascular smooth muscle cells and tissues were examined. Analysis with a confocal laser microscope showed that DC-A (0.1-30 microM) permeabilized the plasma membrane of A10 cells to the non-permeable fluorescent agents, ... More
Increased production of tumor necrosis factor-alpha by glial cells exposed to simulated ischemia or elevated hydrostatic pressure induces apoptosis in cocultured retinal ganglion cells.
AuthorsTezel G, Wax MB
JournalJ Neurosci
PubMed ID11102475
'Although glial cells in the optic nerve head undergo a reactivation process in glaucoma, the role of glial cells during glaucomatous neurodegeneration of retinal ganglion cells is unknown. Using a coculture system in which retinal ganglion cells and glial cells are grown on different layers but share the same culture ... More
Thrombin receptor activation protects neurons and astrocytes from cell death produced by environmental insults.
'Thrombin is a multifunctional serine protease that is rapidly produced from prothrombin at sites of tissue injury and catalyzes the final steps in blood coagulation. Thrombin also regulates gene expression and process outgrowth in neurons and astrocytes and stimulates proliferation of astrocytes. Since thrombin is produced immediately upon breakdown of ... More
Use of thiazole orange homodimer as an alternative to ethidium bromide for DNA detection in agarose gels.
AuthorsWilke WW, Heller MJ, Iakoubova OK, Robinson RA
JournalMod Pathol
PubMed ID8058712
'Detection of polymerase chain reaction-amplified DNA fragments is commonly accomplished by visualizing the products in electrophoretic agarose beds with the use of ethidium bromide under ultraviolet light. However, ethidium bromide is mutagenic, and special handling and disposal precautions must be used. We report the use of a nonmutagenic dye, thiazole ... More
Mitochondrial hyperpolarization after transient oxygen-glucose deprivation and subsequent apoptosis in cultured rat hippocampal neurons.
AuthorsIijima T, Mishima T, Akagawa K, Iwao Y
JournalBrain Res
PubMed ID14642839
'Mitochondrial membrane potential (MMP) regulates the production of high-energy phosphate and apoptotic cascade, both occurring after ischemic impact. The timed profile of MMP differing from grading ischemic impact has to be determined. Primary rat hippocampal cultures were exposed to oxygen-glucose deprivation (OGD) for 30, 60, and 90 min and then ... More
Synthesis of a Kunitz-type serine proteinase inhibitory protein that shares homology with bovine pancreatic trypsin inhibitor by ovine intervertebral disc cells in serum-free alginate bead culture.
AuthorsMelrose J, Smith S, Ghosh P
JournalJ Spinal Disord Tech
PubMed ID11927828
'The objective of this study was to determine whether disc cells could be cultured under serum-free conditions and whether they synthesized bovine pancreatic trypsin inhibitor (BPTI)-like serine proteinase inhibitory proteins (SPIs) previously demonstrated for ovine chondrocytes. Intervertebral discs from 1- to 2-year-old merino wether sheep were dissected into the annulus ... More
A simple photometric detection method for HLA-DRB1 specific PCR-SSP products.
AuthorsFerencik S, Grosse-Wilde H
JournalEur J Immunogenet
PubMed ID8494870
Use of fluorescent probes to investigate the metabolic state of Pneumocystis carinii mitochondria.
AuthorsChen F, Cushion MT
JournalJ Eukaryot Microbiol
PubMed ID7804275
Ethidium dimer: a new reagent for the fluorimetric determination of nucleic acids.
AuthorsMarkovits J, Roques BP, Le Pecq JB
JournalAnal Biochem
PubMed ID464292
Assays for testing Pneumocystis carinii viability.
AuthorsKaneshiro ES, Wu YP, Cushion MT
JournalJ Protozool
PubMed ID1726329
A series of classical vital stains and fluorescent indicator compounds were evaluated as viability assays of P. carinii. The combination of the acetoxymethyl ester of calcein with either ethidium homodimer or propidium iodide distinguished between live, dead and moribund organisms and provided high fluorescence intensity and low bleaching enabling photodocumentation ... More
Direct detection of PCR products for HLA class II typing.
AuthorsChia D, Terasaki P, Chan H, Tonai R, Siauw PA
JournalTissue Antigens
PubMed ID8284789
Direct detection of the PCR, or DD-PCR is proposed as an efficient method for performing PCR assays. Following the PCR reaction, ethidium homodimer dye is added to the reaction mixture and read by fluorescence. The dye step circumvents the necessity of running reactions on agarose gel electrophoresis, which is the ... More
The use of fluorescent nuclear dyes for the study of blood vessel structure and function: novel applications of existing techniques.
AuthorsDaly CJ, Gordon JF, McGrath JC
JournalJ Vasc Res
PubMed ID1372834
We have used nuclear fluorescent dyes to develop a technique for the study of vascular structure and function. Nuclear stained blood vessels, viewed with the appropriate filter sets, can be studied in great detail. Only the nuclei of the cells which form the walls are visible and so their positions ... More
Intracellular trehalose improves the survival of cryopreserved mammalian cells.
AuthorsEroglu A, Russo MJ, Bieganski R, Fowler A, Cheley S, Bayley H, Toner M
JournalNat Biotechnol
PubMed ID10657121
We report that the introduction of low concentrations of intracellular trehalose can greatly improve the survival of mammalian cells during cryopreservation. Using a genetically engineered mutant of Staphylococcus aureus alpha-hemolysin to create pores in the cellular membrane, we were able to load trehalose into cells. Low concentrations (0.2 M) of ... More
Trehalose expression confers desiccation tolerance on human cells.
AuthorsGuo N, Puhlev I, Brown DR, Mansbridge J, Levine F
JournalNat Biotechnol
PubMed ID10657122
Many organisms that withstand desiccation express the disaccharide trehalose. We have now expressed the otsA and otsB genes of Escherichia coli, which encode trehalose biosynthetic enzymes, in human primary fibroblasts using a recombinant adenovirus vector. Infected cells produced increased amounts of trehalose with increasing multiplicity of infection (MOI). Human primary ... More
Effect of B-Z transition and nucleic acid structure on the conformational dynamics of bound ethidium dimer measured by hydrogen deuterium exchange kinetics.
AuthorsMarkovits J, Ramstein J, Roques BP, Le Pecq JB
JournalNucleic Acids Res
PubMed ID4011442
Ethidium dimer is shown to bind by intercalation, almost equally well, to the B and Z form of poly[(dG-m5dC)].poly[(dG-m5dC)], whereas the ethidium monomer shows a strong preference for the B form. The hydrogen-deuterium (H-D) exchange kinetics of the ethidium dimer bound to the B and Z form of poly [(dG-m5dC)].poly[(dG-m5dC)] ... More
Adenovirus E1A represses cardiac gene transcription and reactivates DNA synthesis in ventricular myocytes, via alternative pocket protein- and p300-binding domains.
AuthorsKirshenbaum LA, Schneider MD
JournalJ Biol Chem
PubMed ID7713869
To examine the potential impact of disrupting "pocket" protein function on cardiac differentiation and growth, we introduced 12 S E1A genes into neonatal ventricular myocytes, by adenoviral gene transfer. In the absence of E1B, E1A was cytotoxic, with features typical of apoptosis. In the presence of E1B, E1A preferentially inhibited ... More
Dynamic structure of DNA complexes. Fluorometric measurement of hydrogen-deuterium exchange kinetics of dna-bound ethidium dimer and acridine-ethidium dimer.
AuthorsMarkovits J, Ramstein J, Roques BP, Le Pecq JB
JournalBiochemistry
PubMed ID6882747
The hydrogen-deuterium (H-D) exchange kinetics of free and DNA-bound ethidium dimer and acridine-ethidium heterodimer were measured by stopped flow using fluorescence detection. This technique allowed a very accurate measurement of the exchange process. The H-D exchange kinetics were measured in various environments. In some cases, it was observed that the ... More
Novel protein transfection of primary rat cortical neurons using an antibody that penetrates living cells.
AuthorsWeisbart RH, Baldwin R, Huh B, Zack DJ, Nishimura R
JournalJ Immunol
PubMed ID10820286
An Ab-based system to deliver functional proteins into neurons was developed using the murine mAb, mAb 3E10. This was achieved by covalently conjugating catalase to the Ab so that the conjugate retained high activity for the degradation of hydrogen peroxide. Three-dimensional fluorescence microscopy was used to demonstrate penetration of the ... More
Inhibition of xenoreactive natural antibody production by retroviral gene therapy.
AuthorsBracy JL, Sachs DH, Iacomini J
JournalScience
PubMed ID9743496
The major barrier to transplantation across discordant species, such as from pig to human, is rejection mediated by xenoreactive natural antibodies (XNA) that bind the carbohydrate epitope Galalpha1-3Galbeta1-4GlcNAc-R (alphaGal) on donor tissues. This epitope is synthesized by the enzyme glucosyltransferase uridine 5'-diphosphate galactose:beta-D-galactosyl-1, 4-N-acetyl-D-glucosaminide alpha(1-3)galactosyltransferase (E.C. 2.4.1.151), or simply alphaGT. ... More
Transverse agarose pore gradient gel electrophoresis of DNA.
AuthorsFawcett JS, Wheeler D, Chrambach A
JournalJ Biochem Biophys Methods
PubMed ID1640052
Transverse agarose pore gradient gels were prepared on GelBond in the concentration range of nominally 0.2-1.5% SeaKem GTG agarose, using density stabilization by glycerol and incorporation of a dye to define the gel concentration at each point on the pore gradient gel. The distribution of the dye was evaluated by ... More
Optimal processing method to obtain four-color confocal fluorescent images of the cytoskeleton and nucleus in three-dimensional chondrocyte cultures.
AuthorsBlanc A, Tran-Khanh N, Filion D, Buschmann MD
JournalJ Histochem Cytochem
PubMed ID15933071
Tissue engineering of articular cartilage requires accurate imaging of the chondrocyte cytoskeleton. Past studies have applied various fixation and permeabilization protocols without optimization of parameters. In this study, we have examined procedures using glutaraldehyde and paraformaldehyde as fixatives and Triton X-100 and Octyl-POE as permeabilizing detergents. A four-color fluorescence confocal ... More
Improved fluorescent bioassay for the detection of tumor necrosis factor activity.
AuthorsLévesque A, Paquet A, Pagé M
JournalJ Immunol Methods
PubMed ID7829867
Tumor necrosis factor alpha (TNF-alpha) is a monokine of 17 kDa produced by activated macrophages and various cells involved in the immune system. We propose a new method for the measurement of TNF activity on mouse L929 fibroblast cells. After an incubation with TNF, the cells were stained with a ... More
In vitro toxicity testing of supramolecular sensitizers for photodynamic therapy.
We report the phototoxicity of meso-tetrakis(4-sulphonatophenyl)porphine (TPPS4) and zinc metallocomplex (ZnTPPS4) sensitizers in the presence or absence of 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD) on G361human melanoma cells. Morphological changes in cell cultures have been evaluated using inversion fluorescent microscope and image analysis. Viability of cells was determined by means of molecular probes for ... More
UVA-induced apoptosis studied by the new apo/necro-Comet-assay which distinguishes viable, apoptotic and necrotic cells.
AuthorsMorley N, Rapp A, Dittmar H, Salter L, Gould D, Greulich KO, Curnow A,
JournalMutagenesis
PubMed ID16500949
An adaptation of the Comet-assay was developed which enables the discrimination of viable, apoptotic and necrotic single cells by use of the common Annexin-V staining and a dye exclusion test on the cells already embedded in agarose gel on glass slides. Membrane integrity (Ethidium-Homodimer exclusion), cellular esterase activity (Calcein blue-AM) ... 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
Rnd3 regulation of the actin cytoskeleton promotes melanoma migration and invasive outgrowth in three dimensions.
AuthorsKlein RM, Aplin AE,
JournalCancer Res
PubMed ID19244113
The depth of cell invasion into the dermis is a clinical determinant for poor prognosis in cutaneous melanoma. The signaling events that promote the switch from a noninvasive to invasive tumor phenotype remain obscure. Activating mutations in the serine/threonine kinase B-RAF are prevalent in melanoma. Mutant B-RAF is required for ... More
Potentiometric platform for the quantification of cellular potassium efflux.
AuthorsGenerelli S, Jacquemart R, de Rooij NF, Jolicoeur M, Koudelka-Hep M, Guenat OT,
JournalLab Chip
PubMed ID18584100
Renewed interest in the measurement of cellular K(+) effluxes has been prompted by the observation that potassium plays an active and important role in numerous key cellular events, in particular cell necrosis and apoptosis. Although necrosis and apoptosis follow different pathways, both induce intracellular potassium effluxes. Here, we report the ... More
Cartilage tissue engineering: Directed differentiation of embryonic stem cells in three-dimensional hydrogel culture.
AuthorsHwang NS, Varghese S, Elisseeff J,
JournalMethods Mol Biol
PubMed ID18453267
The clinical goal of tissue engineering is to restore, repair, or replace damaged tissues in the body. Significant advances have been made in recent years using stem cells as a cell source for cartilage tissue engineering and reconstructive surgery applications. Embryonic stem cells have demonstrated the potential to self-renew and ... More
Neurotoxic potential of depleted uranium effects in primary cortical neuron cultures and in Caenorhabditis elegans.
AuthorsJiang GC, Tidwell K, McLaughlin BA, Cai J, Gupta RC, Milatovic D, Nass R, Aschner M,
JournalToxicol Sci
PubMed ID17636247
Depleted uranium (DU) is an extremely dense metal that is used in radiation shielding, counterbalances, armor, and ammunition. In light of the public concerns about exposure to DU and its potential role in Gulf War Syndrome (GWS), this study evaluated the neurotoxic potential of DU using focused studies on primary ... More
Microfluidic-based cell sorting of Francisella tularensis infected macrophages using optical forces.
AuthorsPerroud TD, Kaiser JN, Sy JC, Lane TW, Branda CS, Singh AK, Patel KD,
JournalAnal Chem
PubMed ID18510341
We have extended the principle of optical tweezers as a noninvasive technique to actively sort hydrodynamically focused cells based on their fluorescence signal in a microfluidic device. This micro fluorescence-activated cell sorter (microFACS) uses an infrared laser to laterally deflect cells into a collection channel. Green-labeled macrophages were sorted from ... More
Functional protein delivery into neurons using polymeric nanoparticles.
AuthorsHasadsri L, Kreuter J, Hattori H, Iwasaki T, George JM,
JournalJ Biol Chem
PubMed ID19129199
An efficient route for delivering specific proteins and peptides into neurons could greatly accelerate the development of therapies for various diseases, especially those involving intracellular defects such as Parkinson disease. Here we report the novel use of polybutylcyanoacrylate nanoparticles for delivery of intact, functional proteins into neurons and neuronal cell ... More
Effect of vitamin B6 on oxygen radicals, mitochondrial membrane potential, and lipid peroxidation in H2O2-treated U937 monocytes.
AuthorsKannan K, Jain SK
JournalFree Radic Biol Med
PubMed ID14975445
Vitamin B6 (Vit.B6) supplementation has been shown to be beneficial in reducing diabetic complications, cognitive aging, and in the prevention of coronary heart disease. It was hypothesized that Vit.B6 compounds may function as antioxidants and thus offer protection against oxidative stress under various pathophysiological and or experimental conditions. To test ... More
Glutamate-induced neuronal death: a succession of necrosis or apoptosis depending on mitochondrial function.
AuthorsAnkarcrona M, Dypbukt JM, Bonfoco E, Zhivotovsky B, Orrenius S, Lipton SA, Nicotera P
JournalNeuron
PubMed ID7576644
During ischemic brain injury, glutamate accumulation leads to overstimulation of postsynaptic glutamate receptors with intracellular Ca2+ overload and neuronal cell death. Here we show that glutamate can induce either early necrosis or delayed apoptosis in cultures of cerebellar granule cells. During and shortly after exposure to glutamate, a subpopulation of ... More
Cu(II) potentiation of alzheimer abeta neurotoxicity. Correlation with cell-free hydrogen peroxide production and metal reduction.
AuthorsHuang X, Cuajungco MP, Atwood CS, Hartshorn MA, Tyndall JD, Hanson GR, Stokes KC, Leopold M, Multhaup G, Goldstein LE, Scarpa RC, Saunders AJ, Lim J, Moir RD, Glabe C, Bowden EF, Masters CL, Fairlie DP, Tanzi RE, Bush AI
JournalJ Biol Chem
PubMed ID10601271
Oxidative stress markers as well as high concentrations of copper are found in the vicinity of Abeta amyloid deposits in Alzheimer's disease. The neurotoxicity of Abeta in cell culture has been linked to H(2)O(2) generation by an unknown mechanism. We now report that Cu(II) markedly potentiates the neurotoxicity exhibited by ... More
Quantification of neurotoxicity and identification of cellular subsets in a three-dimensional brain model.
AuthorsPulliam L, Stubblebine M, Hyun W
JournalCytometry
PubMed ID9581626
Imaging of cells in a large intact three-dimensional tissue remains difficult. Quantification and identification of cell damage in a mixed culture system has been limited by the inability of fluorescent probes to discriminate types of cellular death and penetrate tissue more that 100 microm thick. We have investigated several probes ... More