Search results for "Neutral Red"
showing 10 items of 30 documents
In vitro effect of cadmium and copper on separated blood leukocytes of Dicentrarchus labrax.
2013
The immunotoxic effects of heavy metals on blood leukocytes of sea bass (Dicentrarchus labrax) were examined. The cells, separated by a discontinuous Percoll-gradients, were exposed in vitro to various sublethal concentrations of cadmium and copper (10(-7) M, 10(-5) M, and 10(-3) M) and their immunotoxic effect was then evaluated by measuring neutral red uptake, MU assay, DNA fragmentation and Hsp70 gene expression. First of all, we demonstrated that the cells treated in vitro could incorporate Cd and Cu. A relationship between heavy metal exposure and dose-time-dependent alterations in responses of leukocytes from blood was found for both metals, but copper was more immunotoxic than cadmiu…
Biological and analytical studies of peritoneal dialysis solutions
2018
The purpose of our work was to conduct biological and analytical studies of the peritoneal dialysis (PD) solutions containing glucose and sodium lactate and establish correlations between cell viability of the Vero cell line and values of analytical indexes of the tested solutions. The results of this study confirm the cytotoxicity of the PD solutions even compared with the isotonic solution of sodium chloride, which may be due to the low pH of the solutions, presence of glucose degradation products (GDPs) and high osmolarity of the solutions, and unphysiological concentrations of glucose and sodium lactate. However, it is not yet known what factors or their combination and to what extent c…
Stable expression of human cytochrome P450 1A1 cDNA in V79 Chinese hamster cells and metabolic activation of benzo[a]pyrene
1993
A V79 Chinese hamster cell line stably expressing human cytochrome P450 1A1 (CYP1A1) was obtained by chromosomal integration of the human CYP1A1 cDNA under the control of the SV40 early promoter. Chromosomal integration was verified by Southern analysis, and effective transcription of the human CYP1A1 cDNA was demonstrated by Northern analysis. The CYP1A1 cDNA-encoded protein was characterized by Western analysis using anti-rat CYP1A1. Intracellular association of CYP1A1 with the endoplasmic reticulum could be visualized by in situ immunofluorescence. Crude cell lysates of the V79 derived cell line was able to catalyze 7-ethoxyresorufin-O-deethylation (EROD) with an activity of about 50 pmo…
Evaluation of the cytotoxic effect of 7keto-stigmasterol and 7keto-cholesterol in human intestinal (Caco-2) cells
2012
The biological implications of cholesterol oxidation products have been investigated, though research on plant sterol oxidation products is scarce and in some cases contradictory. The cytotoxicity of 7keto(k)-stigmasterol versus 7keto(k)-cholesterol at different concentrations (0-120 μM) and incubation times (4-24h), in intestinal epithelial cells (Caco-2 cells) was evaluated. The 3-[4,5-dimethylthiazol-2-yl]-2,3-diphenyl tetrazolium bromide and neutral red uptake tests, mitochondrial membrane potential (ΔΨm), and relative DNA and RNA contents in the cell cycle phases were determined. Possible interaction effects between 7k-derivatives or non-oxidized stigmasterol were monitored. Endo/lysos…
Uridine uptake inhibition as a cytotoxicity test for a human hepatoma cell line (HepG2 cells): comparison with the neutral red assay
2001
International audience; This study describes a sensitive microassay for measuring cytotoxicity based on the degree of inhibition of RNA synthesis in HepG2 cells. RNA synthesis is measured by the kinetic uptake of radiolabeled uridine. A large number of compounds were tested in a wide range of concentrations. The concentration required to induce 50% inhibition of HepG2 uridine uptake rates (IC50) was determined for each compound and used to rank its potency. These IC50s were compared with IC50s measured with the neutral red assay. 2-acetylaminofluorene, benzo[a]pyrene and methylnitrosourea were not cytotoxic in the neutral red assay. Uridine uptake was always inhibited at lower concentrations…
Comparative analysis of eight cytotoxicity assays evaluated within the ACuteTox Project.
2013
A comparative analysis of eight cytotoxicity assays [the 3T3 and normal human keratinocytes Neutral Red Uptake (NRU) assay, the primary rat hepatocytes, human HepG2 and 3T3 MU assay, and the human A.704, SH-SY5Y and HepG2 cells propidium iodide (PI) assay] included in several work packages of the EU Integrated Project ACuteTox, has been carried out. The aim was to evaluate whether cells originating from liver, kidney and brain provided different in vitro acute toxicity results, and the influence of primary liver cells versus cell lines originated from the same tissue. Spearman rank correlation analysis and Hierarchical Cluster Analysis were performed based on the IC50 (50% inhibitory concen…
Effect of in vitro exposure to cadmiumand copper on sea bass blood cells
2009
Blood cells freshly collected from sea bass (Dicentrarchus labrax) were exposed in vitro to different concentrations of cadmium (Cd) and copper (Cu) at 10-7 M, 10-5 M, 10-3 M, and exam- ined for neutral red retention capacity and for cell vitality with MTT assay. A relationship between heavy metal exposure and alteration in responses of blood cells in a dose-time-dependent was found. Our results showed that fish blood cells may constitute an interesting biological model for experimen- tal and applied toxicology, especially in the case of environmental pollution.
Formation of fumonisin B(1)-glucose reaction product, in vitro cytotoxicity, and lipid peroxidation on kidney cells.
2010
Fumonisin B(1) (FB(1)) content in corn products decreases during the heating process in foods containing reducing sugars, mainly because of the formation of N-(carboxymethyl)fumonisin B(1). In this study, a rapid method has been developed for the determination of both compounds in corn products using a high-speed blender, Ultra-Turrax, for solvent extraction and liquid chromatography-tandem mass spectrometry. The kinetics of FB(1) degradation and the formation of the Maillard adduct were studied in a model system constituted by corn bread spiked with FB(1) and heated at 160, 180, and 200 degrees C for 3, 6, 10, 15, and 20 min. FB(1) decreased from 0.96 to 0.3 mg/kg and N-(carboxymethyl)fumo…
Influence of cadmium on the morphology and functionality of haemocytes in the compound ascidian Botryllus schlosseri
2013
In order to get insights into the effects of cadmium (Cd) on cell morphology and functions, we exposed haemocytes of the colonial ascidian Botryllus schlosseri to sub-lethal concentrations of CdCl(2). Results indicate that Cd hampers haemocyte spreading and phagocytosis in a dose-dependent way, through the alteration of the actin cytoskeleton. In addition, the metal decreases the stability of the internal membranes, as revealed by the Neutral Red assay. The fraction of cells showing positivity for the lysosomal enzyme acid phosphatase is also reduced in the presence of Cd, whereas the number of cells responsive to the Annexin-V assay and showing chromatin condensation increases, suggesting …
Supravital Uptake of Cationic Dyes by Mast Cell Granules: A Light and Electron Microscope Study
1994
Methylene blue and neutral red were selected for staining mast cell granules by supravital injections. A new technique was applied for embedding in paraffin and Araldite without dislocation or loss of dye. Stabilization and electron microscopic identification of the dyes were achieved by transforming them into electron-dense precipitates using phosphomolybdic acid dissolved in a paraformaldehyde-glutaraldehyde mixture to preserve the ultrastructure of the tissues. It was found that in general the intensity of the light microscopic staining correlated directly with the electron density. Closer study revealed that not all cytoplasmic granules exhibited the same strong affinity for the cationi…