Mitochondrial oxidative phosphorylation (OXPHOS) is central to physiology and disease pathogenesis. To systematically investigate its activity and regulation, we performed a wide range of assays of OXPHOS physiology and nuclear and mitochondrial gene expression across 2490 chemical perturbations in muscle cells. Through mining of the resulting compendium, we discovered that: ... More
Modification of the cytoplasmic domain of influenza virus hemagglutinin affects enlargement of the fusion pore.
AuthorsKozerski C,Ponimaskin E,Schroth-Diez B,Schmidt MF,Herrmann A
JournalJournal of virology
PubMed ID10906206
The fusion activity of chimeras of influenza virus hemagglutinin (HA) (from A/fpv/Rostock/34; subtype H7) with the transmembrane domain (TM) and/or cytoplasmic tail (CT) either from the nonviral, nonfusogenic T-cell surface protein CD4 or from the fusogenic Sendai virus F-protein was studied. Wild-type or chimeric HA was expressed in CV-1 cells ... More
Authors
Journal
PubMed ID10891486
Cellular alterations produced by the experimental increase in intracellular calcium and the nature of protective effects from pretreatment with nimodipine.
The immortalized septal cell line, SN56 B5 G4, generated by the fusion of mouse septal area cells and neuroblastoma cells, was used to determine if nimodipine, an antagonist of voltage sensitive calcium 'L' channels, might act in a neuroprotective fashion when intracellular calcium levels were raised by incubation in ouabain ... 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 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
Control of integrin alphaIIb beta3 outside-in signaling and platelet adhesion by sensing the physical properties of fibrin(ogen) substrates.
AuthorsPodolnikova NP, Yermolenko IS, Fuhrmann A, Lishko VK, Magonov S, Bowen B, Enderlein J, Podolnikov AV, Ros R, Ugarova TP,
JournalBiochemistry
PubMed ID19929007
The physical properties of substrates are known to control cell adhesion via integrin-mediated signaling. Fibrin and fibrinogen, the principal components of hemostatic and pathological thrombi, may represent biologically relevant substrates whose variable physical properties control adhesion of leukocytes and platelets. In our previous work, we have shown that binding of ... More
Adhesion and cytosolic dye transfer between macrophages and intestinal epithelial cells.
AuthorsMartin CA, el-Sabban ME, Zhao L, Burakoff R, Homaidan FR
JournalCell Adhes Commun
PubMed ID9638330
Activated macrophages (M phi) found in the intestinal lesions of patients with inflammatory bowel disease (IBD) secrete many inflammatory mediators which can regulate intestinal epithelial cell (IEC) function. However, little is known about direct M phi-IEC interactions. Two potential mechanisms by which cells may interact are through specific receptor-ligand binding ... More
Activation of monocytes by HIV-Tat treatment is mediated by cytokine expression.
AuthorsLafrenie RM, Wahl LM, Epstein JS, Yamada KM, Dhawan S
JournalJ Immunol
PubMed ID9378998
Treatment of primary monocytes with soluble HIV-Tat protein is associated with increased monocyte metalloproteinase-9 (MMP-9) expression and enhanced beta 2 integrin expression that increases monocyte/endothelial adhesion. These alterations require greater than 12 h of HIV-Tat treatment, suggesting the involvement of intermediate factors. Thus, we have examined the role of cytokines ... More
Cytokine-regulated expression of E-selectin, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) in human microvascular endothelial cells.
AuthorsHaraldsen G, Kvale D, Lien B, Farstad IN, Brandtzaeg P
JournalJ Immunol
PubMed ID8786319
Endothelial cells (EC) recruit circulating leukocytes to sites of inflammation, partly by expression of endothelial-leukocyte adhesion molecules. Whereas the regulation of some adhesion molecules is well characterized in cultured HUVEC, similar data for microvascular human test systems are limited. We studied the cytokine-regulated expression of vascular cell adhesion molecules E-selectin, ... More
Calcein: a novel marker for lymphocytes which enter lymph nodes.
AuthorsWeston SA, Parish CR
JournalCytometry
PubMed ID1451604
Previous studies have identified unique cell surface antigens which are associated with the specific binding of lymphocytes to high endothelial venules (HEV). Evidence is presented in this paper which demonstrates that uptake of the fluorescent dye calcein by lymphocytes represents an additional marker for the lymph node homing subpopulation of ... More
ClC-3 is a fundamental molecular component of volume-sensitive outwardly rectifying Cl- channels and volume regulation in HeLa cells and Xenopus laevis oocytes.
AuthorsHermoso M, Satterwhite CM, Andrade YN, Hidalgo J, Wilson SM, Horowitz B, Hume JR
JournalJ Biol Chem
PubMed ID12183454
Volume-sensitive osmolyte and anion channels (VSOACs) are activated upon cell swelling in most vertebrate cells. Native VSOACs are believed to be a major pathway for regulatory volume decrease (RVD) through efflux of chloride and organic osmolytes. ClC-3 has been proposed to encode native VSOACs in Xenopus laevis oocytes and in ... 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
Rapid assessment of physiological status in Escherichia coli using fluorescent probes.
AuthorsPorter J, Edwards C, Pickup RW
JournalJ Appl Bacteriol
PubMed ID7592133
'Rapid and direct viability assessment of Escherichia coli in filtered, sterile lake water was possible using multiparameter flow cytometry. Fluorescent dyes were used as probes for different cellular functions (membrane potential, membrane integrity and intracellular enzyme activity), which were correlated with the ability of the cells to respond to nutrient ... More
Caspase activation contributes to delayed death of heat-stressed striatal neurons.
AuthorsWhite MG, Emery M, Nonner D, Barrett JN
JournalJ Neurochem
PubMed ID14622126
'Hyperthermia can contribute to brain damage both during development and post-natally. We used rat embryonic striatal neurons in culture to study mechanisms underlying hyperthermia-induced neuronal death. Heat stress at 43 degrees C for 2 h produced no obvious signs of damage during the first 12 h after the stress, but ... More
Overexpression of Na(+)/K (+)-ATPase parallels the increase in sodium transport and potassium recycling in an in vitro model of proximal tubule cellular ageing.
AuthorsSilva E, Gomes P, Soares-da-Silva P,
JournalJ Membr Biol
PubMed ID17334838
'Na(+)/K(+)-ATPase plays a key role in the transport of Na(+) throughout the nephron, but ageing appears to be accompanied by changes in the regulation and localization of the pump. In the present study, we examined the effect of in vitro cell ageing on the transport of Na(+) and K(+) ions ... More
Selective cell targeting with light-absorbing microparticles and nanoparticles.
AuthorsPitsillides CM, Joe EK, Wei X, Anderson RR, Lin CP
JournalBiophys J
PubMed ID12770906
'We describe a new method for selective cell targeting based on the use of light-absorbing microparticles and nanoparticles that are heated by short laser pulses to create highly localized cell damage. The method is closely related to chromophore-assisted laser inactivation and photodynamic therapy, but is driven solely by light absorption, ... 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
Distinct Intracellular Signaling in Tumor Necrosis Factor-related Apoptosis-inducing Ligand- and CD95 Ligand-mediated Apoptosis.
Authors Velthuis Jurjen H L; Rouschop Kasper M A; De Bont Hans J G M; Mulder Gerard J; Nagelkerke J Fred;
JournalJ Biol Chem
PubMed ID11980895
'Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potent inducer of apoptosis in tumor cells but not in healthy cells. Similar to CD95 ligand (CD95L), TRAIL signaling requires ligand-receptor interaction; the downstream signaling molecules, such as Fas-associated death domain and caspase-8, also seem similar. Using cells stably expressing TRAIL and ... More
A seed for Alzheimer amyloid in the brain.
AuthorsHayashi H, Kimura N, Yamaguchi H, Hasegawa K, Yokoseki T, Shibata M, Yamamoto N, Michikawa M, Yoshikawa Y, Terao K, Matsuzaki K, Lemere CA, Selkoe DJ, Naiki H, Yanagisawa K
JournalJ Neurosci
PubMed ID15152051
'A fundamental question about the early pathogenesis of Alzheimer''s disease (AD) concerns how toxic aggregates of amyloid beta protein (Abeta) are formed from its nontoxic soluble form. We hypothesized previously that GM1 ganglioside-bound Abeta (GAbeta) is involved in the process. We now examined this possibility using a novel monoclonal antibody ... 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
Overexpression of the ferritin iron-responsive element decreases the labile iron pool and abolishes the regulation of iron absorption by intestinal epithelial (Caco-2) cells.
AuthorsGárate MA, Núñez MT
JournalJ Biol Chem
PubMed ID10636858
'Mammalian cells regulate iron levels tightly through the activity of iron-regulatory proteins (IRPs) that bind to RNA motifs called iron-responsive elements (IREs). When cells become iron-depleted, IRPs bind to IREs present in the mRNAs of ferritin and the transferrin receptor, resulting in diminished translation of the ferritin mRNA and increased ... More
Increased apical insertion of the multidrug resistance protein 2 (MRP2/ABCC2) in renal proximal tubules following gentamicin exposure.
'Multidrug resistance protein (MRP) 2 (MRP2; ABCC2), an organic anion transporter apically expressed in liver, kidney, and intestine, plays an important protective role through facilitating the efflux of potentially toxic compounds. We hypothesized that upon a toxic insult, MRP2 is up-regulated in mammalian kidney, thereby protecting the tissue from damage. ... More
Two distinct intracytoplasmic regions of the T-cell adhesion molecule CD28 participate in phosphatidylinositol 3-kinase association.
AuthorsPagès F, Ragueneau M, Klasen S, Battifora M, Couez D, Sweet R, Truneh A, Ward SG, Olive D
JournalJ Biol Chem
PubMed ID8621607
'Through the interaction with its ligands, CD80/B7-1 and CD86/B7-2 or B70, the human CD28 molecule plays a major functional role as a costimulator of T cells along with the CD3-TcR complex. We and others have previously reported that phosphatidylinositol 3-kinase inducibly associates with CD28. This association is mediated by the ... More
Stimulation of beta1-integrin function by epidermal growth factor and heregulin-beta has distinct requirements for erbB2 but a similar dependence on phosphoinositide 3-OH kinase.
AuthorsAdelsman MA, McCarthy JB, Shimizu Y
JournalMol Biol Cell
PubMed ID10473632
'Integrins and growth factor receptors are important participants in cellular adhesion and migration. The EGF receptor (EGFR) family of tyrosine kinases and the beta1-integrin adhesion receptors are of particular interest, given the implication for their involvement in the initiation and progression of tumorigenesis. We used adhesion and chemotaxis assays to ... More
Reduction of connexin43 expression and dye-coupling during neuronal differentiation of human NTera2/clone D1 cells.
AuthorsBani-Yaghoub M, Bechberger JF, Naus CC
JournalJ Neurosci Res
PubMed ID9211986
'Gap junctions are plasma membrane specializations that allow direct communication among adjoining cells. We used a human pluripotential teratocarcinoma cell line, NTera-2/clone D1 (NT2/D1), as a model to study gap junctions in CNS neurons and their neuronal precursors. These cells were differentiated following retinoic acid (RA) treatment for 4 weeks ... More
Cadherins mediate intercellular mechanical signaling in fibroblasts by activation of stretch-sensitive calcium-permeable channels.
AuthorsKo KS, Arora PD, McCulloch CA
JournalJ Biol Chem
PubMed ID11466312
'Cells in mechanically active environments form extensive, cadherin-mediated intercellular junctions that are important in tissue remodeling and differentiation. Currently, it is unknown whether adherens junctions in connective tissue fibroblasts transmit mechanical signals and coordinate multicellular adaptations to physical forces. We hypothesized that cadherins mediate intercellular mechanotransduction by activating calcium-permeable, stretch-sensitive ... More
Analysis of competition binding between soluble and membrane-bound ligands for cell surface receptors.
AuthorsLi P, Selvaraj P, Zhu C
JournalBiophys J
PubMed ID10585962
'Binding of the Fc portion of IgG coated on targets to Fcgamma receptors (e.g., CD16) expressed on leukocytes (i.e., 2D binding) is an initiating step for immune responses such as phagocytosis or antibody-dependent cellular cytotoxicity. In vivo, circulating leukocytes are exposed to plasma IgG. The competition from soluble IgG (i.e., ... More
Src regulates distinct pathways for cell volume control through Vav and phospholipase Cgamma.
AuthorsBarfod ET, Moore AL, Melnick RF, Lidofsky SD
JournalJ Biol Chem
PubMed ID15866884
'Cell volume recovery in response to swelling requires reorganization of the cytoskeleton and fluid efflux. We have previously shown that electrolyte and fluid efflux via K+ and Cl- channels is controlled by swelling-induced activation of phospholipase Cgamma (PLCgamma). Recently, integrin engagement has been suggested to trigger responses to swelling through ... 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
Light controllable siRNAs regulate gene suppression and phenotypes in cells.
AuthorsNguyen QN, Chavli RV, Marques JT, Conrad PG, Wang D, He W, Belisle BE, Zhang A, Pastor LM, Witney FR, Morris M, Heitz F, Divita G, Williams BR, McMaster GK
JournalBiochim Biophys Acta
PubMed ID16497269
'Small interfering RNA (siRNA) is widely recognized as a powerful tool for targeted gene silencing. However, siRNA gene silencing occurs during transfection, limiting its use is in kinetic studies, deciphering toxic and off-target effects and phenotypic assays requiring temporal, and/or spatial regulation. We developed a novel controllable siRNA (csiRNA) that ... More
Complement activation occurs on subendothelial extracellular matrix in vitro and is initiated by retraction or removal of overlying endothelial cells.
AuthorsHindmarsh EJ, Marks RM
JournalJ Immunol
PubMed ID9637530
'Vascular endothelium is continuously exposed to plasma complement, which could generate a potent proinflammatory signal if activated on the vascular wall. Normal endothelium, however, expresses an anti-inflammatory phenotype, which includes resistance to complement fixation. As activated endothelium converts to a proinflammatory phenotype, we investigated the effect of cytokines on endothelial ... More
Imaging hair cell transduction at the speed of sound: dynamic behavior of mammalian stereocilia.
AuthorsFridberger A, Tomo I, Ulfendahl M, Boutet de Monvel J
JournalProc Natl Acad Sci U S A
PubMed ID16446441
'The cochlea contains two types of sensory cells, the inner and outer hair cells. Sound-evoked deflection of outer hair cell stereocilia leads to fast force production that will enhance auditory sensitivity up to 1,000-fold. In contrast, inner hair cells are thought to have a purely receptive function. Deflection of their ... More
A pore-forming toxin interacts with a GPI-anchored protein and causes vacuolation of the endoplasmic reticulum.
AuthorsAbrami L, Fivaz M, Glauser PE, Parton RG, van der Goot FG
JournalJ Cell Biol
PubMed ID9456314
'In this paper, we have investigated the effects of the pore-forming toxin aerolysin, produced by Aeromonas hydrophila, on mammalian cells. Our data indicate that the protoxin binds to an 80-kD glycosyl-phosphatidylinositol (GPI)-anchored protein on BHK cells, and that the bound toxin is associated with specialized plasma membrane domains, described as ... More
Reactive oxygen species (ROS)-induced ROS release: a new phenomenon accompanying induction of the mitochondrial permeability transition in cardiac myocytes.
'We sought to understand the relationship between reactive oxygen species (ROS) and the mitochondrial permeability transition (MPT) in cardiac myocytes based on the observation of increased ROS production at sites of spontaneously deenergized mitochondria. We devised a new model enabling incremental ROS accumulation in individual mitochondria in isolated cardiac myocytes ... More
A role of the Ca2+/Mg2+-dependent endonuclease in apoptosis and its inhibition by Poly(ADP-ribose) polymerase.
AuthorsYakovlev AG, Wang G, Stoica BA, Boulares HA, Spoonde AY, Yoshihara K, Smulson ME
JournalJ Biol Chem
PubMed ID10807908
'Apoptosis is characterized by various cell morphological and biochemical features, one of which is the internucleosomal degradation of genomic DNA. The role of the human chromatin-bound Ca(2+)- and Mg(2+)-dependent endonuclease (CME) DNAS1L3 and its inhibition by poly(ADP-ribosyl)ation in the DNA degradation that accompanies apoptosis was investigated. The nuclear localization of ... More
Human ClC-3 is not the swelling-activated chloride channel involved in cell volume regulation.
AuthorsWeylandt KH, Valverde MA, Nobles M, Raguz S, Amey JS, Diaz M, Nastrucci C, Higgins CF, Sardini A
JournalJ Biol Chem
PubMed ID11278960
'Volume regulation is essential for normal cell function. A key component of the cells'' response to volume changes is the activation of a channel, which elicits characteristic chloride currents (I(Cl, Swell)). The molecular identity of this channel has been controversial. Most recently, ClC-3, a protein highly homologous to the ClC-4 ... More
Ranking of P-glycoprotein substrates and inhibitors by a calcein-AM fluorometry screening assay.
AuthorsTiberghien F, Loor F
JournalAnticancer Drugs
PubMed ID8862725
'In order to compare the capacities of a variety of compounds to interfere with P-glycoprotein (Pgp) function, a novel assay was set up to work on a large screening scale. The model assay measures the capacity of parental sensitive (Par) and multidrug-resistant (MDR) cells to efflux a small fixed amount ... More
Brain-derived neurotrophic factor promotes survival and chemoprotection of human neuroblastoma cells.
AuthorsMiddlemas DS, Kihl BK, Zhou J, Zhu X
JournalJ Biol Chem
PubMed ID10347207
'Brain-derived neurotrophic factor (BDNF) promotes neuronal survival and protection against neuronal damage. We addressed whether BDNF might promote survival and chemoprotection in neuroblastoma (NB) using a drug-sensitive human NB cell line. All-trans-retinoic acid (ATRA) induces a striking phenotypic differentiation of NB1643 cells, and exogenous BDNF treatment promotes survival of these ... More
A fluorescence assay for assessing chelation of intracellular iron in a membrane model system and in mammalian cells.
AuthorsCabantchik ZI, Glickstein H, Milgram P, Breuer W
JournalAnal Biochem
PubMed ID8789722
'Iron chelators are important tools in biochemical studies of iron metabolism and in the therapy of iron overload diseases. Their mode of action is comprised of entry into cells and scavenging intracellular metal, which includes complexation and egress of the complex. Iron is a metal which appears in the cells ... More
Dominant-negative effect on adhesion by myelin Po protein truncated in its cytoplasmic domain.
AuthorsWong MH, Filbin MT
JournalJ Cell Biol
PubMed ID8830780
'The myelin Po protein is believed to hold myelin together via interactions of both its extracellular and cytoplasmic domains. We have already shown that the extracellular domains of Po can interact in a homophilic manner (Filbin, M.T., F.S. Walsh, B.D. Trapp, J.A. Pizzey, and G.I. Tennekoon. 1990. Nature (Lond.). 344:871-872). ... 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
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
Targeted disruption of the LAMA3 gene in mice reveals abnormalities in survival and late stage differentiation of epithelial cells.
AuthorsRyan MC, Lee K, Miyashita Y, Carter WG
JournalJ Cell Biol
PubMed ID10366601
'Laminin 5 regulates anchorage and motility of epithelial cells through integrins alpha6beta4 and alpha3beta1, respectively. We used targeted disruption of the LAMA3 gene, which encodes the alpha3 subunit of laminin 5 and other isoforms, to examine developmental functions that are regulated by adhesion to the basement membrane (BM). In homozygous ... More
'BACKGROUND: Transient (low-conductance) opening of the mitochondrial permeability transition pore (mPTP) may limit mitochondrial calcium load and mediate mitochondrial reactive oxygen species (ROS) signaling. We hypothesize that transient mPTP opening and ROS mediate the protection associated with myocardial preconditioning and mitochondrial uncoupling. METHODS AND RESULTS: Isolated perfused rat hearts were ... 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
Hyaluronan synthase elevation in metastatic prostate carcinoma cells correlates with hyaluronan surface retention, a prerequisite for rapid adhesion to bone marrow endothelial cells.
'Bone marrow is the primary site of metastasis in patients with advanced stage prostate cancer. Prostate carcinoma cells metastasizing to bone must initially adhere to endothelial cells in the bone marrow sinusoids. In this report, we have modeled that interaction in vitro using two bone marrow endothelial cell (BMEC) lines ... More
Regulatory volume decrease and intracellular Ca2+ in murine neuroblastoma cells studied with fluorescent probes.
'The possible role of Ca2+ as a second messenger mediating regulatory volume decrease (RVD) in osmotically swollen cells was investigated in murine neural cell lines (N1E-115 and NG108-15) by means of novel microspectrofluorimetric techniques that allow simultaneous measurement of changes in cell water volume and [Ca2+]i in single cells loaded ... More
Role of poly(ADP-ribose) polymerase (PARP) cleavage in apoptosis. Caspase 3-resistant PARP mutant increases rates of apoptosis in transfected cells.
AuthorsBoulares AH, Yakovlev AG, Ivanova V, Stoica BA, Wang G, Iyer S, Smulson M
JournalJ Biol Chem
PubMed ID10438458
'An early transient burst of poly(ADP-ribosyl)ation of nuclear proteins was recently shown to be required for apoptosis to proceed in various cell lines (Simbulan-Rosenthal, C., Rosenthal, D., Iyer, S., Boulares, H., and Smulson, M. (1998) J. Biol. Chem. 273, 13703-13712) followed by cleavage of poly(ADP-ribose) polymerase (PARP), catalyzed by caspase-3. ... More
Nramp 2 (DCT1/DMT1) expressed at the plasma membrane transports iron and other divalent cations into a calcein-accessible cytoplasmic pool.
AuthorsPicard V, Govoni G, Jabado N, Gros P
JournalJ Biol Chem
PubMed ID10942769
'Nramp2, also known as DMT1 and DCT1, is a 12-transmembrane (TM) domain protein responsible for dietary iron uptake in the duodenum and iron acquisition from transferrin in peripheral tissues. Nramp2/DMT1 produces by alternative splicing two isoforms differing at their C terminus (isoforms I and II). The subcellular localization, mechanism of ... More
Simultaneous binding of two different drugs in the binding pocket of the human multidrug resistance P-glycoprotein.
AuthorsLoo TW, Bartlett MC, Clarke DM
JournalJ Biol Chem
PubMed ID12909621
'The human multidrug resistance P-glycoprotein (P-gp, ABCB1) transports a wide variety of structurally diverse compounds out of the cell. The drug-binding pocket of P-gp is located in the transmembrane domains. Although occupation of the drug-binding pocket by one molecule is sufficient to activate the ATPase activity of P-gp, the drug-binding ... More
Correlated three-dimensional light and electron microscopy reveals transformation of mitochondria during apoptosis.
AuthorsSun MG, Williams J, Munoz-Pinedo C, Perkins GA, Brown JM, Ellisman MH, Green DR, Frey TG
JournalNat Cell Biol
PubMed ID17721514
'In addition to their role in cellular bioenergetics, mitochondria also initiate common forms of programmed cell death (apoptosis) through the release of proteins such as cytochrome c from the intermembrane and intracristal spaces. The release of these proteins is studied in populations of cells by western blotting mitochondrial and cytoplasmic ... More
Cell volume measured by total internal reflection microfluorimetry: application to water and solute transport in cells transfected with water channel homologs.
AuthorsFarinas J, Simanek V, Verkman AS
JournalBiophys J
PubMed ID7540430
'Total internal reflection (TIR) microfluorimetry was established as a method to measure continuously the volume of adherent cells and applied to measure membrane permeabilities in cells transfected with water channel homologs. Cytosol was labeled with the membrane-impermeant fluorophore calcein. Fluorescence was excited by the TIR evanescent field in a thin ... More
In vitro model of the outer blood-retina barrier.
AuthorsSteuer H, Jaworski A, Stoll D, Schlosshauer B
JournalBrain Res Brain Res Protoc
PubMed ID15063838
'The outer blood-retina barrier (BRB) is formed by the retinal pigment epithelium (rpe) and functions similarly to the blood-brain barrier (BBB). In contrast to the BBB, which is composed of a myriad of capillaries, the rpe can in principle be prepared as an intact planar tissue sheet without disruption of ... More
Bystander tumoricidal effect and gap junctional communication in lung cancer cell lines.
AuthorsImaizumi K, Hasegawa Y, Kawabe T, Emi N, Saito H, Naruse K, Shimokata K
JournalAm J Respir Cell Mol Biol
PubMed ID9476907
'Tumor cells expressing the herpes simplex virus-thymidine kinase (HSV-tk) gene become sensitive to ganciclovir (GCV), and the phenomenon by which tumor cells surrounding the HSV-tk expressing cells also become sensitive to GCV is known as the "bystander effect." The purpose of this study was to investigate the bystander effect in ... More
AuthorsCuschleri J, Gourlay D, Garcia I, Jelacic S, Maier RV
JournalShock
PubMed ID12744486
'Endotoxin-induced intercellular adhesion molecule-1 (ICAM-1) and interleukin 8 (IL-8) production in endothelial cells, which is mediated by Toll-receptor signaling, is essential for optimal neutrophil recruitment and migration during sepsis. Endotoxin also causes stress fiber polymerization that has recently been shown to affect intracellular signaling. However, the role of this polymerization ... More
Metabolite of SIR2 reaction modulates TRPM2 ion channel.
AuthorsGrubisha O, Rafty LA, Takanishi CL, Xu X, Tong L, Perraud AL, Scharenberg AM, Denu JM
JournalJ Biol Chem
PubMed ID16565078
'The transient receptor potential melastatin-related channel 2 (TRPM2) is a nonselective cation channel, whose prolonged activation by oxidative and nitrative agents leads to cell death. Here, we show that the drug puromycin selectively targets TRPM2-expressing cells, leading to cell death. Our data suggest that the silent information regulator 2 (Sir2 ... 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
Effect of cytotoxic necrotizing factor-1 on actin cytoskeleton in human monocytes: role in the regulation of integrin-dependent phagocytosis.
'Cytotoxic necrotizing factor-1 (CNF1) is isolated from pathogenic strains of Escherichia coli and catalyzes the activation of Rho GTPases by the deamidation of a glutamine residue. This toxin induces stress fiber formation, cell spreading, and membrane folding and promotes phagocytosis competence in epithelial cells. We show that CNF1 induces morphologic ... More
Glutamate triggers rapid glucose transport stimulation in astrocytes as evidenced by real-time confocal microscopy.
AuthorsLoaiza A, Porras OH, Barros LF
JournalJ Neurosci
PubMed ID12917367
'Glutamate stimulates glycolysis in astrocytes, a phenomenon that couples astrocytic metabolism with neuronal activity. However, it is not known whether glutamate also affects glucose transporter-1 (GLUT1), the transporter responsible for glucose entry into astrocytes. To address this question, two different real-time single-cell hexose uptake assays were applied to cultured hippocampal ... More
Real-time multi-wavelength fluorescence imaging of living cells.
AuthorsMorris SJ
JournalBiotechniques
PubMed ID2331370
'We describe a new real-time fluorescence video microscope design for capturing intensified images of cells containing dual wavelength "ratio" dyes or multiple dyes. The microscope will perform real-time capture of two separate fluorescence emission images simultaneously, improving the time resolution of spatial distribution of fluorescence to video frame rates (30 ... More
The BCR/ABL oncogene alters the chemotactic response to stromal-derived factor-1alpha.
AuthorsSalgia R, Quackenbush E, Lin J, Souchkova N, Sattler M, Ewaniuk DS, Klucher KM, Daley GQ, Kraeft SK, Sackstein R, Alyea EP, von Andrian UH, Chen LB, Gutierrez-Ramos JC, Pendergast AM, Griffin JD
JournalBlood
PubMed ID10590068
'The chemokine stromal-derived factor-1alpha (SDF-1alpha) is a chemoattractant for CD34(+) progenitor cells, in vitro and in vivo. The receptor for SDF-1alpha, CXCR-4, is a 7 transmembrane domain receptor, which is also a coreceptor for human immunodeficiency virus (HIV). Here we show that transformation of hematopoietic cell lines by BCR/ABL significantly ... More
Multidrug transporter activity in lymphocytes.
AuthorsElliott JI, Raguz S, Higgins CF,
JournalBr J Pharmacol
PubMed ID15492020
'Multidrug transporters play a dual role in haematopoietic cells, mediating the efflux of xenobiotics and regulating cell migration. For several reasons including the lack of specific antibodies, reports of multidrug transporter distribution on lymphocytes conflict. Murine B cells have been reported to completely lack transporter activity. Through analysis of parental ... More
The influenza haemagglutinin-induced fusion cascade: effects of target membrane permeability changes.
AuthorsBlumenthal R, Morris SJ
JournalMol Membr Biol
PubMed ID10332736
'To define the stages in influenza haemagglutinin (HA)-mediated fusion the kinetics of fusion between cell pairs consisting of single influenza HA-expressing cells and single erythrocytes (RBC) which had been labelled with both a fluorescent lipid (Dil) in the membrane and a fluorescent solute (calcein) in the aqueous space have been ... More
Fractalkine-mediated endothelial cell injury by NK cells.
AuthorsYoneda O, Imai T, Goda S, Inoue H, Yamauchi A, Okazaki T, Imai H, Yoshie O, Bloom ET, Domae N, Umehara H
JournalJ Immunol
PubMed ID10754298
'Endothelial cells (ECs) are primary targets of immunological attack, and their injury can lead to vasculopathy and organ dysfunction in vascular leak syndrome and in rejection of allografts or xenografts. A newly identified CX3C-chemokine, fractalkine, expressed on activated ECs plays an important role in leukocyte adhesion and migration. In this ... More
Target-cell contact activates a highly selective capacitative calcium entry pathway in cytotoxic T lymphocytes.
AuthorsZweifach A
JournalJ Cell Biol
PubMed ID10662784
'Calcium influx is critical for T cell activation. Evidence has been presented that T cell receptor-stimulated calcium influx in helper T lymphocytes occurs via channels activated as a consequence of depletion of intracellular calcium stores, a mechanism known as capacitative Ca(2+) entry (CCE). However, two key questions have not been ... More
Cell tracing dyes significantly change single cell mechanics.
AuthorsLulevich V, Shih YP, Lo SH, Liu GY,
JournalJ Phys Chem B
PubMed ID19366241
'Cell tracing dyes are very frequently utilized in cellular biology research because they provide highly sensitive fluorescent tags that do not compromise cellular functions such as growth and proliferation. In many investigations concerning cellular adhesion and mechanics, fluorescent dyes have been employed with the assumption of little impact on the ... More
Cryptosporidium parvum: effect of multi-drug reversing agents on the expression and function of ATP-binding cassette transporters.
AuthorsBonafonte MT, Romagnoli PA, McNair N, Shaw AP, Scanlon M, Leitch GJ, Mead JR
JournalExp Parasitol
PubMed ID15172220
'In the present study, the gene expression of three multidrug resistance (MDR) and resistance-associated protein (MRP) transport proteins or efflux pumps was characterized and the phenotypic evidence for such pumps was demonstrated in cultured Madin-Darby canine kidney (MDCK) cells. A gradient for the fluorescent probe calcein was established between parasite ... More
Fibrinogen induces IL-8 synthesis in human neutrophils stimulated with formyl-methionyl-leucyl-phenylalanine or leukotriene B(4).
AuthorsKuhns DB, Nelson EL, Alvord WG, Gallin JI
JournalJ Immunol
PubMed ID11509634
'Human exudative neutrophils have greatly increased stores of the neutrophil chemoattractant IL-8 compared with peripheral blood cells, but the mechanism for the increase is not defined. In this report, we show that treatment of peripheral blood neutrophils with the chemotactic peptide fMLP or with leukotriene B(4) or fibrinogen results in ... More
Membrane topology and glycosylation of the human multidrug resistance-associated protein.
AuthorsBakos E Hegedüs T, Holló Z, Welker E, Tusnády GE, Zaman GJ, Flens MJ, Váradi A, Sarkadi B
JournalJ Biol Chem
PubMed ID8647833
'The membrane topology of the human multidrug resistance-associated protein (MRP) was examined by flow cytometry phenotyping, immunoblotting, and limited proteolysis in drug-resistant human and baculovirus-infected insect cells, expressing either the glycosylated or the underglycosylated forms of this protein. Inhibition of N-linked glycosylation in human cells by tunicamycin did not inhibit ... 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
Hyperthermic treatment of canine tibia through RF inductive heating of an intramedullary nail: a new experimental approach to hyperthermia for metastatic bone tumours.
AuthorsIkenaga M, Ohura K, Kotoura Y, Yamamuro T, Nakamura T, Oka M, Hiraoka M, Abe M
JournalInt J Hyperthermia
PubMed ID7963807
'Although the incidence of metastatic bone tumours is recently increasing, the local control rate of conventional treatment modalities is not satisfactory. If an intramedullary nail for the fixation of a weakened bone with metastatic lesions can be used as a heat-generating material for hyperthermia, the treatment result is expected to ... More
Nucleic acid binding agents exert local toxic effects on neurites via a non-nuclear mechanism.
AuthorsPin S, Chen H, Lein PJ, Wang MM
JournalJ Neurochem
PubMed ID16441515
'The mechanism by which drugs that target nucleic acids cause neurotoxicity is not well described. We characterized the neurotoxicity of Hoechst 33342 (bis-benzimide), a common cell permeable nuclear dye, in primary neuronal cultures. The mechanism of cell death was not apoptotic, as death is rapid, not accompanied by typical nuclear ... More
Cross-talk between calpain and caspase proteolytic systems during neuronal apoptosis.
AuthorsNeumar RW, Xu YA, Gada H, Guttmann RP, Siman R
JournalJ Biol Chem
PubMed ID12576481
'Cross-talk between calpain and caspase proteolytic systems has complicated efforts to determine their distinct roles in apoptotic cell death. This study examined the effect of overexpressing calpastatin, the specific endogenous calpain inhibitor, on the activity of the two proteolytic systems following an apoptotic stimulus. Human SH-SY5Y neuroblastoma cells were stably ... More
pABC11 (also known as MOAT-C and MRP5), a member of the ABC family of proteins, has anion transporter activity but does not confer multidrug resistance when overexpressed in human embryonic kidney 293 cells.
AuthorsMcAleer MA, Breen MA, White NL, Matthews N
JournalJ Biol Chem
PubMed ID10438534
'Several members of the ABC family of proteins have been implicated in multidrug resistance associated with cancer therapies. A novel member of this gene family, designated pABC11, has been identified using degenerate polymerase chain reaction. The full-length cDNA spans 5881 base pairs and encodes an open reading frame of 1437 ... More
Failure of P-glycoprotein (MDR1) expressed in Xenopus oocytes to produce swelling-activated chloride channel activity.
AuthorsMorin XK, Bond TD, Loo TW, Clarke DM, Bear CE
JournalJ Physiol
PubMed ID7473231
'1. P-glycoprotein, the protein product of the multidrug resistance (MDR1) gene, has ATP-dependent transporter activity. It has been suggested that P-glycoprotein may also function as a volume-regulated chloride channel or chloride channel regulator. To assess the chloride channel function of P-glycoprotein, we examined swelling-activated chloride conductances in Xenopus oocytes injected ... More
In vitro migratory capacity of CD34+ cells is related to hematopoietic recovery after autologous stem cell transplantation.
AuthorsVoermans C, Kooi ML, Rodenhuis S, van der Lelie H, van der Schoot CE, Gerritsen WR
JournalBlood
PubMed ID11157500
'To investigate whether the migratory ability of peripheral blood-derived CD34+ cells of patients undergoing autologous peripheral blood stem cell transplantation is related to the homing efficiency of these cells, the migration in vitro of these cells was determined and correlated with in vivo hematopoietic recovery. Large inter-individual differences of the ... More
Rapid assessment of p-glycoprotein-drug interactions at the blood-brain barrier.
AuthorsBubik M, Ott M, Mahringer A, Fricker G
JournalAnal Biochem
PubMed ID16942742
'We report on a cost- and time-effective fluorescent dye (calcein-acetoxymethylester; calcein-AM)-based assay to screen compounds for p-glycoprotein (p-gp) interactions at the blood-brain barrier. This assay enables the use of freshly isolated or freshly defrosted endothelial cells without prior cell culture. Isolated porcine brain capillary endothelial cells (PBCECs) in suspension were ... More
cAMP-Dependent potentiation of the Ca(2+)-activated release of the anionic fluorescent dye, calcein, from rat parotid acinar cells.
AuthorsSugita M, Hirono C, Furuya K, Yamagishi S, Kanno Y, Shiba Y
JournalEur J Pharmacol
PubMed ID10675730
'A recent study indicates that elevation of [Ca(2+)](i) enhances the release of calcein, an anionic fluorescent dye, from isolated exocrine acinar cells, so cytoplasmic calcein is useful for monitoring the secretion of organic anions. In this study, we investigated the effect of cAMP on the calcein release evoked by elevation ... More
The multidrug resistance-associated protein (MRP) is over-expressed and functional in rat hepatoma cells.
AuthorsPayen L, Courtois A, Vernhet L, Guillouzo A, Fardel O
JournalInt J Cancer
PubMed ID10209965
'Expression of multidrug-resistance-associated protein (MRP), a drug efflux pump transporting a wide range of xenobiotics, including anti-cancer drugs and chemical carcinogens, and present at low levels in normal hepatocytes, was investigated in rat hepatoma cells. Northern-blot analysis allowed detection of high levels of MRP mRNA in rat diethylnitrosamine-induced hepatocarcinomas when ... More
Serum amyloid P component forms a stable complex with human C5b6.
AuthorsBarbashov SF, Wang C, Nicholson-Weller A
JournalJ Immunol
PubMed ID9103450
'A 30,000 m.w. protein bound tightly to C5b6, which was formed by activating C7-depleted human serum with zymosan. The protein remained bound to the C5b6 complex during the isolation procedure for C5b6, including chromatography on lysine-Sepharose and an anion exchange resin. Following electrophoresis and electroblotting of the C5b6 complex to ... More
Specific single or double proline substitutions in the "spring-loaded" coiled-coil region of the influenza hemagglutinin impair or abolish membrane fusion activity.
AuthorsQiao H, Pelletier SL, Hoffman L, Hacker J, Armstrong RT, White JM
JournalJ Cell Biol
PubMed ID9628890
'We tested the role of the "spring-loaded" conformational change in the fusion mechanism of the influenza hemagglutinin (HA) by assessing the effects of 10 point mutants in the region of high coiled-coil propensity, HA2 54-81. The mutants included proline substitutions at HA2 55, 71, and 80, as well as a ... More
Compound profiling for P-glycoprotein at the blood-brain barrier using a microplate screening system.
AuthorsBauer B, Miller DS, Fricker G
JournalPharm Res
PubMed ID12948014
'PURPOSE: The purpose of this study was to establish a fluorescent dye (calcein-acetoxymethylester; calcein-AM)-based assay to rapidly screen compounds for interactions with p-glycoprotein (p-gp) at the blood-brain barrier and to determine whether such an assay can be useful for kinetic analysis. METHODS: Porcine brain capillary endothelial cells (PBCECs) were isolated ... 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
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
Optimal inhibition of X4 HIV isolates by the CXC chemokine stromal cell-derived factor 1 alpha requires interaction with cell surface heparan sulfate proteoglycans.
AuthorsValenzuela-Fernández A, Palanche T, Amara A, Magerus A, Altmeyer R, Delaunay T, Virelizier JL, Baleux F, Galzi JL, Arenzana-Seisdedos F
JournalJ Biol Chem
PubMed ID11352904
'The chemokine stromal cell-derived factor 1 (SDF-1) is the natural ligand for CXC chemokine receptor 4 (CXCR4). SDF-1 inhibits infection of CD4+ cells by X4 (CXCR4-dependent) human immunodeficiency virus (HIV) strains. We previously showed that SDF-1 alpha interacts specifically with heparin or heparan sulfates (HSs). Herein, we delimited the boundaries ... More
Droplet microfluidic technology for single-cell high-throughput screening.
AuthorsBrouzes E, Medkova M, Savenelli N, Marran D, Twardowski M, Hutchison JB, Rothberg JM, Link DR, Perrimon N, Samuels ML,
JournalProc Natl Acad Sci U S A
PubMed ID19617544
'We present a droplet-based microfluidic technology that enables high-throughput screening of single mammalian cells. This integrated platform allows for the encapsulation of single cells and reagents in independent aqueous microdroplets (1 pL to 10 nL volumes) dispersed in an immiscible carrier oil and enables the digital manipulation of these reactors ... More
Identification of protein targets of 4-hydroxynonenal using click chemistry for ex vivo biotinylation of azido and alkynyl derivatives.
AuthorsVila A, Tallman KA, Jacobs AT, Liebler DC, Porter NA, Marnett LJ,
JournalChem Res Toxicol
PubMed ID18232660
'Polyunsaturated fatty acids (PUFA) are primary targets of free radical damage during oxidative stress. Diffusible electrophilic alpha,beta-unsaturated aldehydes, such as 4-hydroxynonenal (HNE), have been shown to modify proteins that mediate cell signaling (e.g., IKK and Keap1) and alter gene expression pathways responsible for inducing antioxidant genes, heat shock proteins, and ... More
Cyclic adenosine monophosphate is a key component of regulatory T cell-mediated suppression.
AuthorsBopp T, Becker C, Klein M, Klein-Hessling S, Palmetshofer A, Serfling E, Heib V, Becker M, Kubach J, Schmitt S, Stoll S, Schild H, Staege MS, Stassen M, Jonuleit H, Schmitt E,
JournalJ Exp Med
PubMed ID17502663
'Naturally occurring regulatory T cells (T reg cells) are a thymus-derived subset of T cells, which are crucial for the maintenance of peripheral tolerance by controlling potentially autoreactive T cells. However, the underlying molecular mechanisms of this strictly cell contact-dependent process are still elusive. Here we show that naturally occurring ... More
Mitochondrial permeability transition and swelling can occur reversibly without inducing cell death in intact human cells.
AuthorsMinamikawa T, Williams DA, Bowser DN, Nagley P
JournalExp Cell Res
PubMed ID9882512
'Severe disruption of mitochondrial function is generally considered to provide a powerful trigger for apoptosis in mammalian cells. We report here that intact cells may undergo the mitochondrial permeability transition and mitochondria swell in a fully reversible manner, without inducing cell death. Cultured human osteosarcoma cells (143B TK-) stained with ... More
Guidelines for the use and interpretation of assays for monitoring cell death in higher eukaryotes.
AuthorsGalluzzi L, Aaronson SA, Abrams J, Alnemri ES, Andrews DW, Baehrecke EH, Bazan NG, Blagosklonny MV, Blomgren K, Borner C, Bredesen DE, Brenner C, Castedo M, Cidlowski JA, Ciechanover A, Cohen GM, De Laurenzi V, De Maria R, Deshmukh M, Dynlacht BD, El-Deiry WS, Flavell RA, Fulda S, Garrido C, Golstein P, Gougeon ML, Green DR, Gronemeyer H, Hajnóczky G, Hardwick JM, Hengartner MO, Ichijo H, Jäättelä M, Kepp O, Kimchi A, Klionsky DJ, Knight RA, Kornbluth S, Kumar S, Levine B, Lipton SA, Lugli E, Ma
JournalCell Death Differ
PubMed ID19373242
'Cell death is essential for a plethora of physiological processes, and its deregulation characterizes numerous human diseases. Thus, the in-depth investigation of cell death and its mechanisms constitutes a formidable challenge for fundamental and applied biomedical research, and has tremendous implications for the development of novel therapeutic strategies. It is, ... More
Fluorescence analysis of the labile iron pool of mammalian cells.
AuthorsEpsztejn S, Kakhlon O, Glickstein H, Breuer W, Cabantchik I
JournalAnal Biochem
PubMed ID9177722
'The labile iron pool (LIP) of cells constitutes a cytosolic fraction of iron which is accessible to permeant chelators and contains the cells' metabolically and catalytically reactive iron. LIP is maintained by a balanced movement of iron from extra- and intracellular sources. We describe here an approach for tracing LIP ... More
Methods for the assessment of mitochondrial membrane permeabilization in apoptosis.
AuthorsGalluzzi L, Zamzami N, de La Motte Rouge T, Lemaire C, Brenner C, Kroemer G
JournalApoptosis
PubMed ID17294081
'Mitochondrial membrane permeabilization (MMP) is considered as the "point-of-no-return" in numerous models of programmed cell death. Indeed, mitochondria determine the intrinsic pathway of apoptosis, and play a major role in the extrinsic route as well. MMP affects the inner and outer mitochondrial membranes (IM and OM, respectively) to a variable ... More
Modeling of P-glycoprotein-involved epithelial drug transport in MDCK cells.
AuthorsIto S, Woodland C, Sarkadi B, Hockmann G, Walker SE, Koren G
JournalAm J Physiol
PubMed ID10409301
'P-glycoprotein (P-gp) on the apical membranes of epithelial cells is known as a drug efflux pump. However, unclear is its integral quantitative role in the overall epithelial drug transfer, which also involves distinct diffusion processes in parallel and sequence. We used a simple three-compartment model to obtain kinetic parameters of ... More