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The uptake and subcellular partitioning of benzo[a]pyrene (BaP) were examined in a rat-liver cell line (Clone 9) using confocal and multiphoton microscopy. Following a 16-h treatment, intracellular accumulation of BaP increased with increasing concentration, and cytoplasmic BaP fluorescence reached saturation at 10 microM. Analysis of the kinetics of BaP(More)
Kinetic analyses of the mechanisms of patulin- and gossypol-induced cellular toxicity in an immortalized rat hepatocyte cell line were examined using a battery of vital fluorescence bioassays. Intracellular glutathione (GSH) content and intracellular Ca2+ ([Ca2+])i were monitored simultaneously using fluorescent probes requiring uv excitation (351-363 nm);(More)
This study examined the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) and related compounds on the uptake of intracellular calcium ([Ca2+]i) in primary cultures of rat hippocampal neuronal cells. [Ca2+]i levels were detected and quantified by interactive laser cytometry with microscopic image analysis. Cells were noninvasively labeled with(More)
The epidermal growth factor receptor (EGFR), which regulates cell growth and survival, is integral to colon tumorigenesis. Lipid rafts play a role in regulating EGFR signaling, and docosahexaenoic acid (DHA) is known to perturb membrane domain organization through changes in lipid rafts. Therefore, we investigated the mechanistic link between EGFR function(More)
Astroglia serve as a presumptive lead (Pb) sink in the brain; therefore, this study examined Pb entry into cultured rat astroglia utilizing the Ca2+ fluorophore indo-1 as a tool for detecting Pb2+ entry during acute exposure. The interactions of Pb2+ with indo-1 were analyzed by fluorescence spectrophotometry in a cell-free system. The emission spectrum of(More)
This paper reports the results from in vitro experiments utilizing vital fluorescent probes and biochemical assays to examine the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) and related compounds in primary rat astroglia in an effort to identify the cellular site(s) involved in toxicity. Application of 100 nM 2,3,7,8-TCDD, a strong Ah(More)
We have shown that a combination of fish oil (high in n-3 fatty acids) with the butyrate-producing fiber pectin, upregulates apoptosis in colon cells exposed to the carcinogen azoxymethane, protecting against colon tumor development. We now hypothesize that n-3 fatty acids prime the colonocytes such that butyrate can initiate apoptosis. To test this, 30(More)
Docosahexaenoic acid (DHA, 22:6 n-3) from fish oil, and butyrate, a fiber fermentation product, work coordinately to protect against colon tumorigenesis in part by inducing apoptosis. We have recently demonstrated that dietary DHA is incorporated into mitochondrial membrane phospholipids, thereby enhancing oxidative stress induced by butyrate metabolism. In(More)
Complex spatiotemporal changes in intracellular Ca2+ were monitored in an immortalized human myometrial cell line (PHM1-41) and first-passage human myometrial cells after oxytocin stimulation (1. 0-1000 nM). Laser cytometry revealed intracellular Ca2+ oscillations in both culture systems starting at 1.0 nM, which were followed by repetitive Ca2+ transients(More)
The effects of lead (0.0, 0.1, or 1 microM) on subcellular sites in primary astroglial cultures were quantitated with the use of vital fluorescent probes (fluorescence bioassays). Evaluation of cellular glutathione levels with monochlorobimane revealed a reduction in glutathione content after only 7 hr of Pb treatment to 77 and 82% of control values for 0.1(More)