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
Authors
Journal
PubMed ID10896672
Detection of low copy numbers of HPV DNA by fluorescent in situ hybridization combined with confocal microscopy as an alternative to in situ polymerase chain reaction.
AuthorsLizard G, Chignol MC, Souchier C, Roignot P, Chardonnet Y, Schmitt D
JournalJ Virol Methods
PubMed ID9672129
'In genital lesions infected by human papillomavirus (HPV), histological criteria and HPV DNA typing are of prognostic value. Therefore, non-radioactive methods such as in situ hybridization are used extensively since they preserve the histological organization of the tissue, and allow the detection and characterization of HPV DNA. However, the sensitivity ... More
Identification of an amino acid residue in multidrug resistance protein 1 critical for conferring resistance to anthracyclines.
AuthorsZhang DW, Cole SP, Deeley RG
JournalJ Biol Chem
PubMed ID11278596
'Murine multidrug resistance protein 1 (mrp1), unlike human MRP1, does not confer resistance to anthracyclines. Previously, we have shown that a human/murine hybrid protein containing amino acids 959-1187 of MRP1 can confer resistance to these drugs. We have now examined the functional characteristics of mutant proteins in which we have ... More
Oxidative stress induces intracellular accumulation of amyloid beta-protein (Abeta) in human neuroblastoma cells.
AuthorsMisonou H, Morishima-Kawashima M, Ihara Y
JournalBiochemistry
PubMed ID10841777
'Several lines of evidence suggest that enhanced oxidative stress is involved in the pathogenesis and/or progression of Alzheimer''s disease (AD). Amyloid beta-protein (Abeta) that composes senile plaques, a major neuropathological hallmark of AD, is considered to have a causal role in AD. Thus, we have studied the effect of oxidative ... More
Age-related macular degeneration. The lipofusion component N-retinyl-N-retinylidene ethanolamine detaches proapoptotic proteins from mitochondria and induces apoptosis in mammalian retinal pigment epithelial cells.
AuthorsSuter M, Remé C, Grimm C, Wenzel A, Jäättela M, Esser P, Kociok N, Leist M, Richter C
JournalJ Biol Chem
PubMed ID11006290
'10-20% of individuals over the age of 65 suffer from age-related macular degeneration (AMD), the leading cause of severe visual impairment in humans living in developed countries. The pathogenesis of this complex disease is poorly understood, and no efficient therapy or prevention exists to date. A precondition for AMD appears ... More
Quantum dot-induced epigenetic and genotoxic changes in human breast cancer cells.
AuthorsChoi AO, Brown SE, Szyf M, Maysinger D,
JournalJ Mol Med
PubMed ID17965848
'The staggering array of nanotechnological products, found in our environment and those applicable in medicine, has stimulated a growing interest in examining their long-term impact on genetic and epigenetic processes. We examined here the epigenomic and genotoxic response to cadmium telluride quantum dots (QDs) in human breast carcinoma cells. QD ... 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
A protective role of the low density lipoprotein receptor-related protein against amyloid beta-protein toxicity.
AuthorsVan Uden E, Sagara Y, Van Uden J, Orlando R, Mallory M, Rockenstein E, Masliah E
JournalJ Biol Chem
PubMed ID10899157
'In order to delineate the neuroprotective role of the low density lipoprotein receptor-related protein (LRP) against amyloid beta-protein toxicity, studies were performed in C6 cells challenged with amyloid beta-protein in the presence or absence of activated alpha(2)-macroglobulin. Toxicity was assessed via two cell viability assays. We found that this endocytic ... More
Cyclopentenone prostaglandins as potential inducers of intracellular oxidative stress.
AuthorsKondo M, Oya-Ito T, Kumagai T, Osawa T, Uchida K
JournalJ Biol Chem
PubMed ID11278531
'In the present study, we find that cyclopentenone prostaglandins (PGs) of the J(2) series, naturally occurring derivatives of PGD(2), are potential inducers of intracellular oxidative stress that mediates cell degeneration. Based on an extensive screening of diverse chemical agents on induction of intracellular production of reactive oxygen species (ROS), we ... More
Apoptotic signaling through the beta -adrenergic receptor. A new Gs effector pathway.
AuthorsGu C, Ma YC, Benjamin J, Littman D, Chao MV, Huang XY
JournalJ Biol Chem
PubMed ID10767282
'Stimulation of beta-adrenergic receptor normally results in signaling by the heterotrimeric G protein G(s), leading to the activation of adenylyl cyclase, production of cAMP, and activation of cAMP-dependent protein kinase (PKA). Here we report that cell death of thymocytes can be induced after stimulation of beta-adrenergic receptor, or by addition ... More
Evidence that collapsin response mediator protein-2 is involved in the dynamics of microtubules.
AuthorsGu Y, Ihara Y
JournalJ Biol Chem
PubMed ID10770920
'Collapsin response mediator protein-2 (CRMP-2) is a member of the CRMP/TOAD/Ulip/DRP family of cytosolic phosphoproteins involved in neuronal differentiation and axonal guidance. CRMP-2 mediates the intracellular response to collapsin 1/semaphorin 3A, a repulsive extracellular guidance cue for axonal outgrowth. The mutation of UNC-33, a Caenorhabditis elegans homolog of CRMP-2, results ... More
Intracellular and extracellular cathepsin B facilitate invasion of MCF-10A neoT cells through reconstituted extracellular matrix in vitro.
AuthorsPremzl A, Zavasnik-Bergant V, Turk V, Kos J
JournalExp Cell Res
PubMed ID12581740
'Lysosomal cysteine proteinase cathepsin B is implicated in remodeling the extracellular matrix, a crucial step in the process of tumor cell invasion. In this study the contributions of intracellular and extracellular cathepsin B activities in the invasion of ras-transformed human breast epithelial cells, MCF-10A neoT, were assessed using specific cathepsin ... More
Secreted beta-amyloid precursor protein counteracts the proapoptotic action of mutant presenilin-1 by activation of NF-kappaB and stabilization of calcium homeostasis.
AuthorsGuo Q, Robinson N, Mattson MP
JournalJ Biol Chem
PubMed ID9575187
'Mutations in the presenilin-1 (PS-1) gene account for approximately 50% of the cases of autosomal dominant, early onset, inherited forms of Alzheimer's disease (AD). PS-1 is an integral membrane protein expressed in neurons and is localized primarily in the endoplasmic reticulum (ER). PS-1 mutations may promote neuronal degeneration by altering ... More
Amyloid beta-peptide induces cell monolayer albumin permeability, impairs glucose transport, and induces apoptosis in vascular endothelial cells.
AuthorsBlanc EM, Toborek M, Mark RJ, Hennig B, Mattson MP
JournalJ Neurochem
PubMed ID9109512
'Amyloid beta-peptide (A beta) is deposited as insoluble fibrils in the brain parenchyma and cerebral blood vessels in Alzheimer''s disease (AD). In addition to neuronal degeneration, cerebral vascular alterations indicative of damage to vascular endothelial cells and disruption of the blood-brain barrier occur in AD. Here we report that A ... More
Cellular features of an apoptotic form of programmed cell death during the development of the ascidian, Boltenia villosa.
AuthorsBates WR
JournalZoolog Sci
PubMed ID15170059
'The phylogenetic position of ascidians near the base of the chordate tree makes them ideal organisms for evolutionary developmental studies of programmed cell death (PCD). In the present study, the following key features of an apoptotic form of PCD are described in Boltenia villosa: fragmentation of DNA, increases in plasma ... More
Insulin receptor substrate 1-induced inhibition of endoplasmic reticulum Ca2+ uptake in beta-cells. Autocrine regulation of intracellular ca2+ homeostasis and insulin secretion.
AuthorsXu GG, Gao ZY, Borge PD, Wolf BA
JournalJ Biol Chem
PubMed ID10364259
'To understand the role of the insulin receptor pathway in beta-cell function, we have generated stable beta-cells (betaIRS1-A) that overexpress by 2-fold the insulin receptor substrate-1 (IRS-1) and compared them to vector-expressing controls. IRS-1 overexpression dramatically increased basal cytosolic Ca2+ levels from 81 to 278 nM, but it did not ... More
The potassium channel opener (-)-cromakalim prevents glutamate-induced cell death in hippocampal neurons.
AuthorsLauritzen I, De Weille JR, Lazdunski M
JournalJ Neurochem
PubMed ID9326286
'(-)-Cromakalim, a typical K+-channel opener, prevents neuronal death induced by either glucose and oxygen privation or by high (100 microM) extracellular glutamate in primary cultures of hippocampus. (-)-Cromakalim has no effect on the earliest events associated with exposure to glutamate. It does not prevent the rapid rise of intracellular Ca2+, ... More
Comparison of MTT and ATP-based assays for the measurement of viable cell number.
AuthorsPetty RD, Sutherland LA, Hunter EM, Cree IA
JournalJ Biolumin Chemilumin
PubMed ID7762413
'Cell viability assays are widely used to assess the effect chemotherapeutic drugs and other agents on cell lines and have shown promise for the prediction of tumour chemosensitivity. In this study we have compared two viability assays using Daudi and CCRF-CEM cell lines over a range of 1500-100,000 cells/well of ... More
Reduced susceptibility of nonobese diabetic mice to TNF-alpha and D-galactosamine-mediated hepatocellular apoptosis and lethality.
'Nonobese diabetic (NOD/LtJ or NOD) mice are resistant to doses of LPS and D-galactosamine that uniformly produce lethality in C57BL/6J (B6) mice (p < 0.01). Liver caspase-3-like activity, serum transaminase levels (both p < 0.05), and the numbers of apoptotic liver nuclei were also reduced in NOD compared with B6 ... More
Localization of a substrate specificity domain in the multidrug resistance protein.
AuthorsStride BD, Cole SP, Deeley RG
JournalJ Biol Chem
PubMed ID10428874
'Multidrug resistance protein (MRP) confers resistance to a number of natural product chemotherapeutic agents. It is also a high affinity transporter of some physiological conjugated organic anions such as cysteinyl leukotriene C(4) and the cholestatic estrogen, 17beta-estradiol 17(beta-D-glucuronide) (E(2)17betaG). We have shown that the murine orthologue of MRP (mrp), unlike ... More
Spectrophotometric determination of clonogenic capacity of leukemic cells in a semisolid microtiter culture system.
AuthorsSchweitzer CM, van de Loosdrecht AA, Jonkhoff AR, Ossenkoppele GJ, Huijgens PC, Dräger AM, Broekhoven MG, Langenhuijsen MM
JournalExp Hematol
PubMed ID8462666
'In this study we describe a semiautomated clonogenic assay in which human leukemic cell lines (U937 and HL60) are cultured in a semisolid 96-well microtiter culture system and clonogenicity is measured spectrophotometrically in a 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT)-assay and sulforhodamine (SRB)-assay. Culture medium with 0.6% (wt/vol) methylcellulose and 10% ... 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
Synthesis and characterization of a novel degradable phosphate-containing hydrogel.
AuthorsWang DA, Williams CG, Li Q, Sharma B, Elisseeff JH
JournalBiomaterials
PubMed ID12834592
'A phosphate-containing and photocrosslinkable polymer, poly(ethylene glycol) di-[ethyl phosphatidyl (ethylene glycol) methacrylate], "PhosPEG-dMA", was synthesized. As a water-soluble macromer, PhosPEG-dMA is suitable for in situ injection and cell-encapsulation by light-induced gelation to produce a novel biocompatible and biodegradable hydrogel for application to cartilage and bone tissue engineering. 1H-NMR, MALDI-TOF mass ... More
A new approach for measurement of cytotoxicity using colorimetric assay.
AuthorsHussain RF, Nouri AM, Oliver RT
JournalJ Immunol Methods
PubMed ID8450240
'Using in vitro established tumour cell lines attempts were made to assess the suitability of tetrazolium salt reduction (MTT) assay to replace the conventional radioactive base techniques for measuring cell proliferation and cell killing. The optimum conditions for MTT loading time, concentration of MTT and the time for colour development ... More
Laboratory determination of chemotherapeutic drug resistance in tumor cells from patients with leukemia, using a fluorometric microculture cytotoxicity assay (FMCA).
AuthorsLarsson R, Kristensen J, Sandberg C, Nygren P
JournalInt J Cancer
PubMed ID1730510
'An automated fluorometric microculture cytotoxicity assay (FMCA) based on the measurement of fluorescence generated from cellular hydrolysis of fluorescein diacetate (FDA) to fluorescein was employed for chemotherapeutic-drug-sensitivity testing of tumor-cell suspensions from patients with leukemia. Fluorescence was linearly related to cell number, and reproducible measurements of drug sensitivity could be ... More
Neurotrophin protection against toxicity induced by low potassium and nitroprusside in cultured cerebellar granule neurons.
AuthorsYu O, Chuang DM
JournalJ Neurochem
PubMed ID8978711
'Long-term survival of cultured rat cerebellar granule neurons requires depolarizing concentrations of potassium (high potassium; 25 mM KCl). A high-potassium culturing condition has been reported to increase the intracellular calcium concentration ([Ca2+]i) and the expression of brain-derived neurotrophic factor (BDNF), which in turn induces the expression of neurotrophin-3 (NT-3) in ... More
Hydrogen peroxide mediates amyloid beta protein toxicity.
AuthorsBehl C, Davis JB, Lesley R, Schubert D
JournalCell
PubMed ID8004671
'Amyloid beta protein (A beta) is a 40-43 amino acid peptide that is associated with plaques in the brains of Alzheimer''s patients and is cytotoxic to cultured neurons. Using both primary central nervous system cultures and clonal cell lines, it is shown that a number of anti-oxidants protect cells from ... More
Reduction of a tetrazolium salt and superoxide generation in human tumor cells (HeLa).
AuthorsBurdon RH, Gill V, Rice-Evans C
JournalFree Radic Res Commun
PubMed ID8397148
'Experiments have been carried out to explore the use of a tetrazolium salt, MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide in the detection of intracellularly generated superoxide in HeLa cells. From the use of a low molecular weight lipophilic mimic of superoxide dismutase, as well as superoxide dismutase, and inhibitors of superoxide dismutase, it ... More
Evidence for a cholesterol transport pathway from lysosomes to endoplasmic reticulum that is independent of the plasma membrane.
AuthorsUnderwood KW, Jacobs NL, Howley A, Liscum L
JournalJ Biol Chem
PubMed ID9461625
'We have studied the movement of low density lipoprotein (LDL)-derived cholesterol in cultured Chinese hamster ovary cells. Our hypothesis is that when LDL cholesterol is effluxed from lysosomes, the bulk of LDL cholesterol is mobilized to the plasma membrane, while another pathway delivers LDL cholesterol from lysosomes to acyl-CoA/cholesterol acyltransferase ... More
Suppression of ceramide-mediated programmed cell death by sphingosine-1-phosphate.
AuthorsCuvillier O, Pirianov G, Kleuser B, Vanek PG, Coso OA, Gutkind S, Spiegel S
JournalNature
PubMed ID8657285
'Ceramide is an important regulatory participant of programmed cell death (apoptosis) induced by tumour-necrosis factor (TNF)-alpha and Fas ligand, members of the TNF superfamily. Conversely, sphingosine and sphingosine-1-phosphate, which are metabolites of ceramide, induce mitogenesis and have been implicated as second messengers in cellular proliferation induced by platelet-derived growth factor ... More
A noninvasive cell-based assay for monitoring proteolytic activity within a specific subcellular compartment.
AuthorsBelkhiri A, Lytvyn V, Guilbault C, Bourget L, Massie B, Nägler DK, Ménard R
JournalAnal Biochem
PubMed ID12123661
'A noninvasive cell-based assay has been developed to monitor the proteolytic activity of cathepsin L within a specific subcellular compartment, the lysosome. The green fluorescent protein (GFP) of Aequorea victoria was selected as a substrate. Targeting to lysosomes was achieved by fusing GFP to preprocathepsin L, which also ensures colocalization ... More
Phosphatidylinositol 3-kinase is a key regulator of early phase differentiation in keratinocytes.
AuthorsSayama K, Yamasaki K, Hanakawa Y, Shirakata Y, Tokumaru S, Ijuin T, Takenawa T, Hashimoto K
JournalJ Biol Chem
PubMed ID12183446
'The survival and growth of epithelial cells depend on adhesion to the extracellular matrix. Because epidermal keratinocytes differentiate as they leave the basement membrane, an adhesion signal may regulate the initiation of differentiation. Phosphatidylinositol 3-kinase (PI3K) is a fundamental signaling molecule that regulates the adhesion signal. Transfection of a dominant ... More
Factors affecting the selection and use of tetrazolium salts as cytochemical indicators of microbial viability and activity.
AuthorsThom SM, Horobin RW, Seidler E, Barer MR
JournalJ Appl Bacteriol
PubMed ID7683637
'The reduction of four tetrazolium cations (TCs), nitroblue tetrazolium (NBT), neotetrazolium (NT), methylthiazolyldiphenyltetrazolium (MTT) and iodonitrophenyltetrazolium (INT), by viable micro-organisms, immobilized on glass cover slips, was examined by light microscopy with a view to determining a systematic basis for applying these reagents as cytochemical indicators of microbial viability and activity. ... More
Biotransformation of the MPTP analog trans-1-methyl-4-[4-dimethylaminophenylethenyl]-1,2,3,6-tetra- hydropyridine to a fluorescent pyridinium metabolite by intact neuroblastoma cells.
AuthorsSong X, Ehrich M, Flaherty D, Wang YX, Castagnoli N
JournalToxicol Appl Pharmacol
PubMed ID8661341
'The possibility of imaging monoamine oxidase (MAO) containing neurons through the MAO-mediated conversion of the nonfluorescent tetrahydropyridine compound trans-1-methyl-4-[4-dimethylaminophenylethenyl]-1,2,3,6-tetrahydro pyridine (t-THP) to the corresponding fluorescent trans-1-methyl-4-[4-dimethylaminophenylethenyl]pyridinium species (t-P+) was examined with the aid of human neuroblastoma cells (SH-SY5Y). Fluorescence microscopy and fluorescence measurements established the intracellular formation of a fluorescent ... More
Inhibition of Akt kinase by cell-permeable ceramide and its implications for ceramide-induced apoptosis.
Authors Zhou H; Summers S A; Birnbaum M J; Pittman R N;
JournalJ Biol Chem
PubMed ID9632728
'Ceramide is an important lipid messenger involved in mediating a variety of cell functions including apoptosis. However, mechanisms responsible for ceramide-induced apoptosis remain unclear. We investigated the possibility that ceramide may decrease antiapoptotic signaling in cells by inhibiting Akt kinase activity. Our data show that C2-ceramide induces apoptosis in HMN1 ... More
In situ assessment of cell viability.
AuthorsYang H, Acker J, Chen A, McGann L
JournalCell Transplant
PubMed ID9786064
'Cryobiological studies of tissues often require the simultaneous assessment of tissue structure and in situ cellular function. Localization of damage during cryopreservation occurs as a consequence of tissue structure and morphology and as a result of biophysical constraints imposed by diffusion and heat transfer. This study used five experimental model ... More
15-deoxy-delta 12, 14-Prostaglandin J2 prevents reactive oxygen species generation and mitochondrial membrane depolarization induced by oxidative stress.
AuthorsGarg TK, Chang JY
JournalBMC Pharmacol
PubMed ID15149553
'BACKGROUND: With the use of cultured human retinal pigment epithelial cells, we have previously described a number of cellular responses to oxidative stress caused by H2O2. We also demonstrated that the cytotoxicity caused by H2O2 could be prevented by the prostaglandin derivative, 15-deoxy-delta 12, 14-Prostaglandin J2 (15d-PGJ2). RESULTS: Further characterization ... More
(R)-alpha-lipoic acid protects retinal pigment epithelial cells from oxidative damage.
AuthorsVoloboueva LA, Liu J, Suh JH, Ames BN, Miller SS
JournalInvest Ophthalmol Vis Sci
PubMed ID16249512
'PURPOSE: To determine whether (R)-alpha-lipoic acid (LA) protects cultured human fetal retinal pigment epithelial (hfRPE) cells against oxidative injury and identify the pathways that may mediate protection. METHODS: Cultured hfRPE cells were pretreated with various concentrations of LA for 14 to 16 hours followed by treatment with a chemical oxidant, ... More
Pseudomonas aeruginosa induces apoptosis in human endothelial cells.
AuthorsValente E, Assis MC, Alvim IM, Pereira GM, Plotkowski MC
JournalMicrob Pathog
PubMed ID11095919
'Pseudomonas aeruginosa has been shown to enter into human endothelial cells in vitro. To ascertain the effects of bacterial intracellular (IC) infection, endothelial cells were exposed to PAK and PAO-1 strains for 1 h and treated with gentamicin in culture medium for different periods. P. aeruginosa induced a significant production ... More
Calcium regulates estrogen increase in permeability of cultured CaSki epithelium by eNOS-dependent mechanism.
AuthorsGorodeski GI
JournalAm J Physiol Cell Physiol
PubMed ID11029297
'Estrogen increases baseline transepithelial permeability across CaSki cultures and augments the increase in permeability in response to hypertonic gradients. In estrogen-treated cells, lowering cytosolic calcium abrogated the hypertonicity-induced augmented increase in permeability and decreased baseline permeability to a greater degree than in estrogen-deprived cells. Steady-state levels of cytosolic calcium in ... More
Microcystin-LR and nodularin induce intracellular glutathione alteration, reactive oxygen species production and lipid peroxidation in primary cultured rat hepatocytes.
AuthorsBouaïcha N, Maatouk I
JournalToxicol Lett
PubMed ID15019088
'Microcystin-LR (MCYST-LR) and nodularin (NOD) produced by cyanobacteria are potent specific hepatotoxins. However, the mechanisms of their hepatotoxicity have not been fully elucidated. In the present study the effect of non cytotoxic low concentrations of MCYST-LR and NOD on intracellular reduced glutathione (GSH) alteration, reactive oxygen species (ROS) production and ... More
A microassay for measuring cytochrome P450IA1 and P450IIB1 activities in intact human and rat hepatocytes cultured on 96-well plates.
AuthorsDonato MT, Gómez-Lechón MJ, Castell JV
JournalAnal Biochem
PubMed ID8238878
'This study deals with the development of a sensitive and simple microassay for measuring cytochrome P450IA1 and P450IIB1 activities in intact human and rat hepatocytes. The method is based on the metabolism by cells cultured in microwells of appropriate substrates at noncytotoxic concentrations (8 microM 7-ethoxyresorufin and 15 microM 7-pentoxyresorufin). ... More
Kinetic characterization of the nitric oxide toxicity for PC12 cells: effect of half-life time of NO release.
AuthorsYamamoto T, Yuyama K, Nakamura K, Kato T, Yamamoto H
JournalEur J Pharmacol
PubMed ID10844095
'We investigated the effects of low concentrations of nitric oxide (NO) on cell viability using NO donors, (+/-)-(E)-4-methyl-2-[(E)-hydroxyimino]-5-nitro-6-methoxy-3-hex enamid e (NOR1), (+/-)-(E)-4-methyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide (NOR2), (+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide (NOR3) and (+/-)-N-[(E)-4-ethyl-2-[(Z)-hydroxyimino]-5-nitro-3-hexen-1- yl]-3-pyr idine (NOR4). The half-life times of the NO release from these four NOR analogs, NOR1, NOR2, NOR3 and NOR4, were determined (6.5, ... More
Oxidative glutamate toxicity involves mitochondrial dysfunction and perturbation of intracellular Ca2+ homeostasis.
AuthorsPereira CF, Oliveira CR
JournalNeurosci Res
PubMed ID10940457
'Glutamate toxicity on PC12 cells is mediated by oxidative stress as a consequence of the inhibition of a cystine uptake system with depletion of GSH. In this study we report that glutamate decreases PC12 cell viability, inhibiting the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). This decrease was prevented by the antioxidants ... More
Cryopreserved human hepatocytes: characterization of drug-metabolizing enzyme activities and applications in higher throughput screening assays for hepatotoxicity, metabolic stability, and drug-drug interaction potential.
AuthorsLi AP, Lu C, Brent JA, Pham C, Fackett A, Ruegg CE, Silber PM
JournalChem Biol Interact
PubMed ID10418968
'Cryopreserved human hepatocytes were extensively characterized in our laboratory. The post-thaw viability, measured via dye exclusion, ranged from 55 to 83%, for hepatocytes cryopreserved from 17 donors. Post-thaw viability and yield (viable cells per vial) were found to be stable up to the longest storage duration evaluated of 120 days. ... More
Degradation studies and biological behavior on an artificial cornea material.
AuthorsSantos L, Ferraz MP, Shirosaki Y, Lopes MA, Fernandes MH, Osaka A, Santos JD,
JournalInvest Ophthalmol Vis Sci
PubMed ID21421875
'Patients with dry eye syndrome, Stevens-Johnson syndrome, or recurrent transplant rejections are unsuitable to receive a keratoprosthesis. The present work aims at developing a highly biocompatible keratoprosthesis that could be successfully implanted in such patients. Glass-reinforced hydroxyapatite (GRHA) was used to construct this new artificial cornea. To grant the device ... More
Fibrillogenesis and cytotoxic activity of the amyloid-forming apomyoglobin mutant W7FW14F.
AuthorsSirangelo I, Malmo C, Iannuzzi C, Mezzogiorno A, Bianco MR, Papa M, Irace G
JournalJ Biol Chem
PubMed ID14701846
'The apomyoglobin mutant W7FW14F forms amyloid-like fibrils at physiological pH. We examined the kinetics of fibrillogenesis using three techniques: the time dependence of the fluorescence emission of thioflavin T and 1-anilino-8-naphthalenesulfonate, circular dichroism measurements, and electron microscopy. We found that in the early stage of fibril formation, non-native apomyoglobin molecules ... More
Comparison of two rapid colorimetric methods for determining resistance of mycobacterium tuberculosis to rifampin, isoniazid, and streptomycin in liquid medium.
AuthorsDe Logu A, Uda P, Pellerano ML, Pusceddu MC, Saddi B, Schivo ML
JournalEur J Clin Microbiol Infect Dis
PubMed ID11245320
'The usefulness of two colorimetric methods for the determination of the susceptibility or resistance of Mycobacterium tuberculosis to rifampin, streptomycin, and isoniazid in liquid medium based on the reduction of 2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide (XTT) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was investigated. The agar proportion method was used as the reference method. Results obtained ... More
Matrix regulation of skeletal cell apoptosis. Role of calcium and phosphate ions.
AuthorsAdams CS, Mansfield K, Perlot RL, Shapiro IM
JournalJ Biol Chem
PubMed ID11278303
'Previously, we noted that inorganic phosphate (P(i)), a major component of bone extracellular matrix, induced osteoblast apoptosis (Meleti, Z., Shapiro, I. M., and Adams, C. S. (2000) Bone (NY) 27, 359-366). Since Ca(2+) along with P(i) is released from bone during the resorption process, we advanced the hypothesis that Ca(2+) ... More
2-substituted 1,2-dihydro-3H-dibenz[de,h]isoquinoline-1,3-diones. A new class of antitumor agent.
AuthorsSami SM, Dorr RT, Alberts DS, Remers WA
JournalJ Med Chem
PubMed ID8459403
'A new class of antitumor agents, having structural analogy to amonafide, but differing by the addition of a fourth ring in the nucleus, was synthesized conveniently from anthracene. Compounds with a variety of substituents, containing a basic nitrogen atom and located on the imide nitrogen, were prepared. Thirteen of 19 ... More
Indomethacin-mediated reversal of multidrug resistance and drug efflux in human and murine cell lines overexpressing MRP, but not P-glycoprotein.
AuthorsDraper MP, Martell RL, Levy SB
JournalBr J Cancer
PubMed ID9062400
'Decreased accumulation of the fluorescent dye BCECF [2'', 7''-bis-(2-carboxyethyl)-5-(6)- carboxyfluorescein] characterized murine and human multidrug-resistant cell lines overexpressing the multidrug resistance protein (MRP). Indomethacin (10 microM), a known cyclo-oxygenase and glutathione-S-transferase inhibitor as well as a modulator of anion transport, increased accumulation and blocked efflux of BCECF in MRP-expressing murine ... More
Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill.
AuthorsHansen MB, Nielsen SE, Berg K
JournalJ Immunol Methods
PubMed ID2470825
'The tetrazolium salt (MTT) method involving conversion of MTT to coloured formazan by cells serving as indirect measurements of cell growth/cell kill has been reported by several groups, although technical problems have been encountered. The present investigation was undertaken in order to delineate what laboratory variables have direct influence on ... More
Replication from oriP of Epstein-Barr virus requires human ORC and is inhibited by geminin.
AuthorsDhar SK, Yoshida K, Machida Y, Khaira P, Chaudhuri B, Wohlschlegel JA, Leffak M, Yates J, Dutta A
JournalCell
PubMed ID11509178
'A hypomorphic mutation made in the ORC2 gene of a human cancer cell line through homologous recombination decreased Orc2 protein levels by 90%. The G1 phase of the cell cycle was prolonged, but there was no effect on the utilization of either the c-Myc or beta-globin cellular origins of replication. ... More
Comparison of antitumor activity of vitamins K1, K2 and K3 on human tumor cells by two (MTT and SRB) cell viability assays.
AuthorsWu FY, Liao WC, Chang HM
JournalLife Sci
PubMed ID8492642
'Vitamin K (VK) congeners (VK1, VK2, and VK3) have been used as antihemorrhagic agents, while VK3 has also been found to inhibit growth in various rodent and human tumor cells. We have compared the antitumor activities of vitamin K1, K2, and K3 against a panel of human cancer cell lines. ... More
Comparison of four different colorimetric and fluorometric cytotoxicity assays in a zebrafish liver cell line.
AuthorsBopp SK, Lettieri T,
JournalBMC Pharmacol
PubMed ID18513395
'BACKGROUND: A broad spectrum of cytotoxicity assays is currently used in the fields of (eco)toxicology and pharmacology. To choose an appropriate assay, different parameters like test compounds, detection mechanism, specificity, and sensitivity have to be considered. Furthermore, tissue or cell line can influence test performance. For zebrafish (Danio rerio), as ... More
Inaccuracies in MTS assays: major distorting effects of medium, serum albumin, and fatty acids.
AuthorsHuang KT, Chen YH, Walker AM
JournalBiotechniques
PubMed ID15470895
'Soluble formazan assays are widely used for cell number assessment. However, in our hands, we observed frequent occasions in which the actual cell number was at odds with the assay reading. In this study, we have determined that (i) a large proportion of the reading obtained in commonly used culture ... More
Tyrphostins protect neuronal cells from oxidative stress.
AuthorsSagara Y, Ishige K, Tsai C, Maher P,
JournalJ Biol Chem
PubMed ID12121989
'Tyrphostins are a family of tyrosine kinase inhibitors originally synthesized as potential anticarcinogenic compounds. Because tyrphostins have chemical structures similar to those of the phenolic antioxidants, we decided to test the protective efficacy of tyrphostins against oxidative stress-induced nerve cell death (oxytosis). Many commercially available tyrphostins, at concentrations ranging from ... More
Augmentation of cytotoxic effect of tumor necrosis factor on human immunodeficiency virus-infected cells by staurosporine, a potent protein kinase C inhibitor.
AuthorsHamamoto Y, Matsuyama T, Yamamoto N, Kobayashi N
JournalCancer Res
PubMed ID2386936
'We have examined the effect of the protein kinase C (PKC) inhibitor, staurosporine, on tumor necrosis factor (TNF)-induced cytotoxic action and augmentation of human immunodeficiency virus (HIV) expression on the chronically HIV-infected T-cell line, MOLT-4/HIV (HTLV-IIIB strain). Staurosporine enhanced the decrease in the number of viable cells caused by TNF ... More
Rapid colorimetric assay for cell viability: application to the quantitation of cytotoxic and growth inhibitory lymphokines.
AuthorsGreen LM, Reade JL, Ware CF
JournalJ Immunol Methods
PubMed ID6609997
'A rapid colorimetric microtiter assay has been developed to detect cytotoxic lymphokines produced by human lymphocytes activated with lectins or tumor cells. The viability of lymphotoxin-treated target cells was detected using a tetrazolium dye that is reduced to a blue formazan by living but not dead cells. The amount of ... More
In the neuronal cell line SH-SY5Y, oxidative stress-induced free radical overproduction causes cell death without any participation of intracellular Ca(2+) increase.
AuthorsAmoroso S, Gioielli A, Cataldi M, Di Renzo G, Annunziato L
JournalBiochim Biophys Acta
PubMed ID10559468
'Adding the membrane-permeant oxidant tert-butylhydroperoxide (t-BOOH) to the incubation medium, in SH-SY5Y human neuroblastoma cells, induced a marked and progressive concentration-dependent (300, 500 and 1000 microM) increase of free radical production, as evaluated by the fluorescent probe 2'',7''-dichlorodihydrofluorescein diacetate (DCFH-DA) and of the intracellular Ca(2+) ion concentrations [Ca(2+)](i). The removal ... More
Quantitative multiwell myeloid differentiation assay using dichlorodihydrofluorescein diacetate (H2DCF-DA) or dihydrorhodamine 123 (H2R123).
AuthorsTrayner ID, Rayner AP, Freeman GE, Farzaneh F
JournalJ Immunol Methods
PubMed ID7594627
'It is well established that the fluorescent probes dichlorodihydrofluorescein diacetate (H2DCF-DA) and dihydrorhodamine 123 (H2R123) can be used to detect the respiratory burst response of mature myeloid cells. We describe a simple, fast and quantitative assay for myeloid differentiation based on the oxidation of these probes, which can be performed ... More
Improved short- and long-term XTT-based colorimetric cellular cytotoxicity assay for melanoma and other tumor cells.
AuthorsJost LM, Kirkwood JM, Whiteside TL
JournalJ Immunol Methods
PubMed ID1548398
'A tetrazolium compound, XTT, bioreducible to a water-soluble formazan was used to develop a simplified cellular cytotoxicity assay. Most (13/15 melanoma and 2/3 colon carcinoma cell lines tested metabolized XTT greater than 50 times more efficiently than the lymphoid effector cells, and thus the test could be performed without separation ... More
Superoxide-mediated decomposition of biological S-nitrosothiols.
AuthorsAleryani S, Milo E, Rose Y, Kostka P
JournalJ Biol Chem
PubMed ID9497319
'Incubation of S-nitrosocysteine or S-nitrosoglutathione (5-100 M) in the presence of a generator of superoxide (xanthine/xanthine oxidase) resulted in a time-dependent decomposition of S-nitrosothiols and accumulation of nitrite/nitrate in reaction mixtures. Quantitatively, the amounts of nitrite/nitrate represented >90% of nitrosonium equivalent of S-nitrosothiols degraded during the incubation. The reaction rates ... More
Falsification of tetrazolium dye (MTT) based cytotoxicity assay results due to mycoplasma contamination of cell cultures.
AuthorsDenecke J, Becker K, Jürgens H, Gross R, Wolff JE
JournalAnticancer Res
PubMed ID10368683
'Mycoplasma contamination of cell cultures is a frequently observed problem. Due to the inconspicuous growth in cell cultures, periodical screening procedures represent the only protection. Many influences of mycoplasma on cell culture parameters have been described. We addressed the question of whether mycoplasma contamination affects the most frequently used cytotoxicity ... More
Fas induces cytoplasmic apoptotic responses and activation of the MKK7-JNK/SAPK and MKK6-p38 pathways independent of CPP32-like proteases.
AuthorsToyoshima F, Moriguchi T, Nishida E
JournalJ Cell Biol
PubMed ID9362518
'IL-1beta converting enzyme (ICE) family cysteine proteases are subdivided into three groups; ICE-, CPP32-, and Ich-1-like proteases. In Fas-induced apoptosis, activation of ICE-like proteases is followed by activation of CPP32-like proteases which is thought to be essential for execution of the cell death. It was recently reported that two subfamily ... More
Nerve growth factor protects against aluminum-mediated cell death.
AuthorsOhyashiki T, Satoh E, Okada M, Takadera T, Sahara M
JournalToxicology
PubMed ID12093616
'In the present study, we examined the effect of two salts of aluminum (Al), aluminum maltolate (Almal) and aluminum chloride (AlCl(3)), on the cell viability of PC12 cells in the absence and presence of nerve growth factor (NGF). A 72-h exposure of PC12 cells to Almal (300 microM) resulted in ... More
Massive programmed cell death in intestinal epithelial cells induced by three-dimensional growth conditions: suppression by mutant c-H-ras oncogene expression.
'Deregulation of molecular pathways controlling cell survival and death, including programmed cell death, are thought to be important factors in tumor formation, disease progression, and response to therapy. Studies devoted to analyzing the role of programmed cell death in cancer have been carried out primarily using conventional monolayer cell culture ... More
Thallium induces hydrogen peroxide generation by impairing mitochondrial function.
AuthorsHanzel CE, Verstraeten SV,
JournalToxicol Appl Pharmacol
PubMed ID16934846
'Thallium (Tl) is highly toxic through yet poorly understood mechanisms. In this study, we comparatively investigated the effects of thallic (Tl(III)) cations on mitochondrial functionality and oxidative stress promotion, and results were compared to those obtained for thallous (Tl(I)) cation. PC12 cells were incubated between 1 and 72 h in ... More
Screening assay for promigratory/antimigratory compounds.
AuthorsRust WL, Huff JL, Plopper GE
JournalAnal Biochem
PubMed ID10805515
'Large-scale screening strategies aimed at finding anticancer drugs traditionally focus on identifying cytotoxic compounds that attack actively dividing cells. Because progression to malignancy involves acquisition of an aggressively invasive phenotype in addition to hyperproliferation, simple and effective screening strategies for finding compounds that target the invasive aspects of cancer progression ... 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
Glutamate receptors induce a burst of superoxide via activation of nitric oxide synthase in arginine-depleted neurons.
AuthorsCulcasi M, Lafon-Cazal M, Pietri S, Bockaert J
JournalJ Biol Chem
PubMed ID7513691
'We have previously shown in cultured cerebellar granule neurons (Lafon-Cazal, M., Pietri, S., Culcasi, M., and Bockaert, J. (1993) Nature 364, 535-537) that upon N-methyl-D-aspartate stimulation, a nitric oxide synthase (NOS)-independent, arachidonic acid-dependent generation of superoxide free radicals (O2-.) is observed after a lag time of 10-15 min. Using the ... More
Protection against hydrogen peroxide-induced cytotoxicity in PC12 cells by scutellarin.
AuthorsHong H, Liu GQ
JournalLife Sci
PubMed ID15051420
'The present study investigated the protective actions of the antioxidant scutellarin against the cytotoxicity produced by exposure to H2O2 in PC12 cells. This was done by assaying for MTT (3,(4,5-dimethylthiazole-2-yl)2,5-diphenyl-tetrazolium bromide) reduction and lactate dehydrogenase (LDH) release. Reactive oxygen species (ROS) and Ca2+ in cells were evaluated by fluorescent microplate ... More
Glutathione depletion in PC12 results in selective inhibition of mitochondrial complex I activity. Implications for Parkinson's disease.
AuthorsJha N, Jurma O, Lalli G, Liu Y, Pettus EH, Greenamyre JT, Liu RM, Forman HJ, Andersen JK
JournalJ Biol Chem
PubMed ID10846169
'Oxidative stress appears to play an important role in degeneration of dopaminergic neurons of the substantia nigra (SN) associated with Parkinson''s disease (PD). The SN of early PD patients have dramatically decreased levels of the thiol tripeptide glutathione (GSH). GSH plays multiple roles in the nervous system both as an ... More
KATP channels regulate mitogenically induced proliferation in primary rat hepatocytes and human liver cell lines. Implications for liver growth control and potential therapeutic targeting.
AuthorsMalhi H, Irani AN, Rajvanshi P, Suadicani SO, Spray DC, McDonald TV, Gupta S
JournalJ Biol Chem
PubMed ID10862612
'To determine whether K(ATP) channels control liver growth, we used primary rat hepatocytes and several human cancer cell lines for assays. K(ATP) channel openers (minoxidil, cromakalim, and pinacidil) increased cellular DNA synthesis, whereas K(ATP) channel blockers (quinidine and glibenclamide) attenuated DNA synthesis. The channel inhibitor glibenclamide decreased the clonogenicity of ... More
Role of cathepsin D in antigen presentation of ovalbumin.
AuthorsRodriguez GM, Diment S
JournalJ Immunol
PubMed ID1328388
'Modification of protein Ag by proteolysis is one of the principal steps in the presentation of Ag to Th cells. However, little is known about the enzymes participating in these events, their specificity or the characteristics of the natural fragments that they produce. Cathepsin D (CD) is an aspartyl protease ... More
Green tea polyphenol epigallocatechin-3-gallate protects HepG2 cells against CYP2E1-dependent toxicity.
AuthorsJimenez-Lopez JM, Cederbaum AI
JournalFree Radic Biol Med
PubMed ID15036355
'Chronic ethanol consumption causes oxidative damage in the liver, and induction of cytochrome P450 2E1 (CYP2E1) is one pathway involved in oxidative stress produced by ethanol. The hepatic accumulation of iron and polyunsaturated fatty acids significantly contributes to ethanol hepatotoxicity in the intragastric infusion model of ethanol treatment. The objective ... More
Comparison of alamar blue and MTT assays for high through-put screening.
AuthorsHamid R, Rotshteyn Y, Rabadi L, Parikh R, Bullock P
JournalToxicol In Vitro
PubMed ID15251189
'The performance of alamar blue and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) cell viability assays in a high through-put format were compared. A total of 117 drugs chosen for their wide range of therapeutic areas were screened at 10 microM using both assays in human hepatoma cell line HepG2. Except for terfenadine ... More
Mercaptoethylguanidine and guanidine inhibitors of nitric-oxide synthase react with peroxynitrite and protect against peroxynitrite-induced oxidative damage.
AuthorsSzabó C, Ferrer-Sueta G, Zingarelli B, Southan GJ, Salzman AL, Radi R
JournalJ Biol Chem
PubMed ID9083027
'Nitric oxide (NO) produced by the inducible nitric-oxide synthase (iNOS) is responsible for some of the pathophysiological alterations during inflammation. Part of NO-related cytotoxicity is mediated by peroxynitrite, an oxidant species produced from NO and superoxide. Aminoguanidine and mercaptoethylguanidine (MEG) are inhibitors of iNOS and have anti-inflammatory properties. Here we ... More
Okadaic acid-induced apoptosis in neuronal cells: evidence for an abortive mitotic attempt.
AuthorsNuydens R, de Jong M, Van Den Kieboom G, Heers C, Dispersyn G, Cornelissen F, Nuyens R, Borgers M, Geerts H
JournalJ Neurochem
PubMed ID9489733
'There is increasing evidence that apoptosis in postmitotic neurons is associated with a frustrated attempt to reenter the mitotic cycle. Okadaic acid, a specific protein phosphatase inhibitor, is currently used in models of Alzheimer''s research to increase the degree of phosphorylation of various proteins, such as the microtubule-associated protein tau. ... More
Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of radiosensitivity.
AuthorsCarmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB
JournalCancer Res
PubMed ID3802101
'Radiation survival curves were generated for V79 Chinese hamster and two human lung cancer cell lines (NCI-H460 and NCI-H249) with doubling times of 10, 20, and 85 h, respectively, using a standard clonogenic assay, a dye exclusion assay, and a semiautomated colorimetric assay utilizing a tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylformazan bromide. Comparable ... More
Differential regulation of mast cell function by IL-10 and stem cell factor.
AuthorsLin TJ, Befus AD
JournalJ Immunol
PubMed ID9378991
'In the development of rodent mast cells (MC), IL-10 significantly enhances the growth factor activity of stem cell factor (SCF). The differential effects of IL-10 and SCF on function of rat peritoneal MC (PMC) are investigated in this study. IL-10 inhibits both constitutive and Ag-induced nitric oxide production by PMC ... More
Different roles for thiol and aspartyl proteases in antigen presentation of ovalbumin.
AuthorsDiment S
JournalJ Immunol
PubMed ID1694878
'By using the model Ag, chicken OVA, the proteolytic events required for effective presentation of the antigenic epitope, OVA323-339 to H-2d-restricted Th cells were investigated. First, the ability of aspartyl and thiol proteases to generate antigenic fragments of Ova in vitro was determined. It was found that cathepsin D, an ... More
Tetrazolium-based cell bioassay for neurotoxins active on voltage-sensitive sodium channels: semiautomated assay for saxitoxins, brevetoxins, and ciguatoxins.
AuthorsManger RL, Leja LS, Lee SY, Hungerford JM, Wekell MM
JournalAnal Biochem
PubMed ID8250223
'In the present study we have developed an assay for the detection of sodium channel-specific marine toxins based upon mitochondrial dehydrogenase activity in the presence of veratridine and ouabain. This cell bioassay allows detection of either sodium channel enhancers, such as the brevetoxins and the ciguatoxins, or sodium channel blocking ... More
Cathepsin B acts as a dominant execution protease in tumor cell apoptosis induced by tumor necrosis factor.
AuthorsFoghsgaard L, Wissing D, Mauch D, Lademann U, Bastholm L, Boes M, Elling F, Leist M, Jäättelä M
JournalJ Cell Biol
PubMed ID11381085
'Death receptors can trigger cell demise dependent or independent of caspases. In WEHI-S fibrosarcoma cells, tumor necrosis factor (TNF) induced an increase in cytosolic cathepsin B activity followed by death with apoptotic features. Surprisingly, this process was enhanced by low, but effectively inhibiting, concentrations of pan-caspase inhibitors. Contrary to caspase ... More
Functional effects of overexpression of protein kinase C-alpha, -beta, -delta, -epsilon, and -eta in the mast cell line RBL-2H3.
'The rat basophilic leukemic (RBL-2H3) cell line was stably transfected with the endogenously expressed Ca2+-dependent protein kinase C-alpha (PKC-alpha) and -betaI and the Ca2+-independent delta and epsilon isoforms to study their functional roles. In addition, the Ca2+-independent PKC-eta was expressed. All transfected PKC isoforms translocated to the membrane-containing fraction in ... More
Multilayered keratinocyte culture used for in vitro toxicology.
AuthorsHoh A, Maier K, Dreher RM
JournalMol Toxicol
PubMed ID2476674
'Human keratinocytes are the most appropriate target cells for evaluating mechanisms of skin cytotoxicity and pharmacology of chemical agents. After having formed a confluent stratified epithelium with proliferating basal cells and differentiated cell layers, human keratinocytes were harvested after enzymatic detachment as stable, three-dimensional cell aggregates or used as adherent ... More
Validation of the MTT dye assay for enumeration of cells in proliferative and antiproliferative assays.
AuthorsLoveland BE, Johns TG, Mackay IR, Vaillant F, Wang ZX, Hertzog PJ
JournalBiochem Int
PubMed ID1417886
'Increasing use is being made of colorimetric assays to quantitate viable cells, e.g., the cellular reduction of the tetrazolium salt, MTT, to formazan by mitochondrial succinate dehydrogenase. We validated this assay for cell proliferation in mixed lymphocyte cultures as compared with 3H-thymidine uptake, and for inhibition of cell proliferation induced ... More
Chloroform Fraction of Centratherum anthelminticum (L.) Seed Inhibits Tumor Necrosis Factor Alpha and Exhibits Pleotropic Bioactivities: Inhibitory Role in Human Tumor Cells.
AuthorsArya A, Achoui M, Cheah SC, Abdelwahab SI, Narrima P, Mohan S, Mustafa MR, Mohd MA,
JournalEvid Based Complement Alternat Med
PubMed ID22474512
'We investigated the antioxidant potential, cytotoxic effect, and TNF-a inhibition activity with NF-?B activation response in a chloroform fraction of Centratherum anthelminticum seeds (CACF). The antioxidant property of CACF was evaluated with DPPH, ORAC, and FRAP assays, which demonstrated significant antioxidant activity. The cytotoxicity of CACF was tested using the ... More
Induction of neuronal cell death by Rab5A-dependent endocytosis of alpha-synuclein.
AuthorsSung JY, Kim J, Paik SR, Park JH, Ahn YS, Chung KC
JournalJ Biol Chem
PubMed ID11316809
'The presynaptic alpha-synuclein is a prime suspect for contributing to Lewy pathology and clinical aspects of diseases, including Parkinson''s disease, dementia with Lewy bodies, and a Lewy body variant of Alzheimer''s disease. Here we examined the pathogenic mechanism of neuronal cell death induced by alpha-synuclein. The exogenous addition of alpha-synuclein ... More
An improved MTT assay using the electron-coupling agent menadione.
AuthorsGarn H, Krause H, Enzmann V, Drössler K
JournalJ Immunol Methods
PubMed ID8308299
'A modification of the MTT based tetrazolium colorimetric assay is described. Using the electron-coupling agent menadione formazan formation by murine splenocytes and P-815 cells was significantly increased whereas dye reduction by macrophages was hardly influenced. These observations suggest that it should be possible to improve the tetrazolium based cytotoxicity assays ... More
Sphingosylphosphorylcholine is a ligand for ovarian cancer G-protein-coupled receptor 1.
AuthorsXu Y, Zhu K, Hong G, Wu W, Baudhuin LM, Xiao Y, Damron DS
JournalNat Cell Biol
PubMed ID10806476
'Sphingosylphosphorylcholine (SPC) is a bioactive lipid that acts as an intracellular and extracellular signalling molecule in numerous biological processes. Many of the cellular actions of SPC are believed to be mediated by the activation of unidentified G-protein-coupled receptors. Here we show that SPC is a high-affinity ligand for an orphan ... More
Identification of bone morphogenetic proteins and their receptors in human breast cancer cell lines: importance of BMP2.
AuthorsArnold SF, Tims E, Mcgrath BE
JournalCytokine
PubMed ID10623428
'The most frequent site of breast cancer metastasis is bone suggesting that some breast cancers express proteins that facilitate this process. We evaluated whether a highly metastatic breast cancer cell line, MDA-MB-231, and a less metastatic breast cancer cell line, MCF-7, contain bone morphogenetic proteins (BMP). Semiquantitative reverse transcriptase-polymerase chain ... More
A simplified new assay for assessment of fungal cell damage with the tetrazolium dye, (2,3)-bis-(2-methoxy-4-nitro-5-sulphenyl)-(2H)-tetrazolium-5-carboxanil ide (XTT).
'Studies of antimycotic host defenses have been limited by the paucity of rapid, reproducible quantitative assays for fungal cell damage. Prior studies defined a colorimetric method that uses MTT, a tetrazolium dye, to quantify polymorphonuclear leukocyte (PMNL)-mediated damage to fungi. These relatively simple, rapid, and reproducible assays require cumbersome extraction ... More
Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays.
AuthorsMosmann T
JournalJ Immunol Methods
PubMed ID6606682
'A tetrazolium salt has been used to develop a quantitative colorimetric assay for mammalian cell survival and proliferation. The assay detects living, but not dead cells and the signal generated is dependent on the degree of activation of the cells. This method can therefore be used to measure cytotoxicity, proliferation ... More
pH dependence and compartmentalization of zinc transported across plasma membrane of rat cortical neurons.
AuthorsColvin RA
JournalAm J Physiol Cell Physiol
PubMed ID11788343
'In this study, Zn(2+) transport in rat cortical neurons was characterized by successfully combining radioactive tracer experiments with spectrofluorometry and fluorescence microscopy. Cortical neurons showed a time-dependent and saturable transport of (65)Zn(2+) with an apparent affinity of 15-20 microM. (65)Zn(2+) transport was pH dependent and was decreased by extracellular acidification ... More
MTT assay overestimates human airway smooth muscle cell number in culture.
AuthorsZhang ZD, Cox G
JournalBiochem Mol Biol Int
PubMed ID8829601
'In this study, we evaluated the accuracy of the MTT (a tetrazolium salt) assay for cell counting by comparing it to haemocytometer. When airway smooth muscle cells were cultured in RPMI with fetal calf serum < or = 5%, no significant differences were observed in cell number counted by the ... More
LTRPC2 Ca2+-permeable channel activated by changes in redox status confers susceptibility to cell death.
AuthorsHara Y, Wakamori M, Ishii M, Maeno E, Nishida M, Yoshida T, Yamada H, Shimizu S, Mori E, Kudoh J, Shimizu N, Kurose H, Okada Y, Imoto K, Mori Y
JournalMol Cell
PubMed ID11804595
'Redox status changes exert critical impacts on necrotic/apoptotic and normal cellular processes. We report here a widely expressed Ca2+-permeable cation channel, LTRPC2, activated by micromolar levels of H2O2 and agents that produce reactive oxygen/nitrogen species. This sensitivity of LTRPC2 to redox state modifiers was attributable to an agonistic binding of ... More
A modified colorimetric MTT assay adapted for primary cultured hepatocytes: application to proliferation and cytotoxicity assays.
AuthorsOka M, Maeda S, Koga N, Kato K, Saito T
JournalBiosci Biotechnol Biochem
PubMed ID1368954
A cytotoxicity assay for tumour necrosis factor using a human rhabdomyosarcoma cell line.