Effect of curcumin on the incorporation of [³H]oleic acid into cholesterylesters in HepG2 cells treated with antipsychotics.

<p>Cells were exposed to LDL (60 μg/ml of cholesterol) in the absence (control) or the presence of haloperidol (Hal), clozapine (Clo), risperidone (Ris), or ziprasidone (Zip) (10 μM) for 16 h. Cells were washed twice and then incubated for additional 2 h in serum-free medium containing the same antipsychotic with or without curcumin (30 μM) and [³H]oleic acid. Then, cells were collected, lipids extracted and separated by HPLC. Results are mean ± SEM of three independent experiments. Statistical comparisons shown antipsychotic versus untreated control (⁺<i>P</i><0.05, ⁺⁺<i>P</i><0.01 and ⁺⁺⁺<i>P</i><0.001).</p

Mamas, Travel Agents and Lot Lizards: Female Gender Stereotypes and Trucking

This presentation, as part of a larger on going study, examines gender stereotypes associated with the occupation of truck driving. According to the U.S. Bureau of Labor Statistics, of the over three million people who make their living as truck drivers, only six percent are female. Using content analysis, gender roles and stereotypes are explored by analyzing a sample of The Trucker, a twice-monthly publication available both online and in print distributed to grocery stores, truck stops and other locations. Preliminary results suggest that women rarely drive solo and are employed most frequently as part of a husband-wife team, or work in dispatch or other office environments. Further, preliminary analysis of photos of women in The Trucker suggest that most women are represented as feminine , while relatively few women are androgynous or masculine in how they are represented

Electron micrographs of HepG2 cells untreated (control) or treated with an antipsychotic (haloperidol or ziprasidone at 10 μM) and LDL (60 μg/ml of cholesterol) for a total of 18 h.

<p>Where indicated, during the last 2 h, the cells were treated with 30 μM curcumin. Images are representative from 2 independent experiments. Lysosomes (▲), lipid inclusions (<b>§</b>), heterolysosomes or MVB (*).</p

Curcumin enhances the release of lysosomal β-hexosaminidase (A) and flotillin-2- and CD63-containing exosomes (B and C) from HepG2 cells treated with antipsychotics.

<p>Cells were exposed to LDL (60 μg/ml of cholesterol) in the absence (control) or the presence of haloperidol (Hal), clozapine (Clo), risperidone (Ris), or ziprasidone (Zip) (10 μM) for 16 h. Cells were washed twice and then incubated for additional 2 h in serum-free medium containing the same antipsychotic with or without curcumin (Cur, 30 μM) as indicated. (A) At the end of the incubation, the activity of the lysosomal enzyme β-hexosaminidase was measured in both cells and medium. The enzyme activity released into the medium is expressed as the percentage of total enzyme content (cells plus medium). Results are mean ± SEM of three independent experiments performed in triplicate. (B and C) At the end of the incubation, the medium was removed, and isolated exosomes and cell lysates analyzed by Western blot. (B) Flotillin-2, CD63, and actin Western blots from exosome and HepG2 cell lysate. Results are representative of 3 independent experiments. (C) Quantitation of exosomal release of flotillin-2 (Flot-2) and CD63 from HepG2 cells. Results are mean ± SEM of three independent experiments; the control without curcumin is normalized to 1. Statistical comparisons shown are curcumin versus no curcumin (* <i>P</i><0.05, ** <i>P</i><0.01, *** <i>P</i><0.001 and **** <i>P</i><0.0001).</p

Curcumin increases LDL-derived cholesterol efflux in HepG2 cells treated with antipsychotics.

<p>HepG2 cells were exposed to ³H-cholesterol-labeled LDL (60 μg/ml of cholesterol) for 16 h in the absence (control) or presence of haloperidol (Hal), clozapine (Clo), risperidone (Ris), or ziprasidone (Zip) (10 μM). Cells were washed twice and serum-free medium was added containing the same antipsychotic with or without 30 μM curcumin, as indicated, and the cells were incubated for additional 2 h. Finally, ³H-radioactivity was measured in both the cells and medium. The ³H-cholesterol efflux into the medium is expressed as the percentage of total radioactivity in the well (cells plus medium). Results are mean ± SEM of three independent experiments performed in duplicate. Statistical comparisons shown are curcumin versus no curcumin (* <i>P</i><0.05, ** <i>P</i><0.01 and *** <i>P</i><0.001).</p

Effects of curcumin on the intracellular distribution of LDL in cells treated with antipsychotics.

<p>(A) HepG2 cells were exposed to DiI-LDL (30 μg/ml of cholesterol) in the absence (control) or the presence of antipsychotic (haloperidol, clozapine, risperidone, or ziprasidone, 10 μM) for a total of 18 h. The cells were then washed, fixed, stained with filipin and anti-CD63, and analyzed by confocal microscopy. Representative results from 3 independent experiments are shown. (B) Cells were exposed to DiI-LDL in the absence (control) or the presence of antipsychotic (10 μM) for a total of 18 h and curcumin (30 μM) was added for the last 2 h. The cells were then washed, fixed, stained with filipin and anti-CD63, and analyzed by confocal microscopy. Representative results from 3 independent experiments are shown.</p