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

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

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

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

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

A putative scheme of PrP^c involvement in oligodendrocyte proliferation and maturation.

<p><b>A-B</b>)<b>.</b> Scheme summarizing the main <i>in vitro</i> findings of the present study. The absence of PrP^c in cultured OPCs from the ON and in the isolated cortical OPCs prolonged the proliferative stage in these precursors, and the modification of intrinsic factors modulating cell proliferation and oligodendrocyte differentiation (e.g., <i>Sox17, cdk2)</i> may delay their normal maturation. <b>C-D</b>) No differences in CNS myelination were observed between <i>Prnp^0/0</i> and <i>Prnp^+/+</i> mice, although increased proliferation was observed in adult NG2 cells in <i>Prnp^0/0</i> mice. The putative changes in intrinsic factors are overcome by extrinsic (neuronal and astroglial) factors to establish normal myelination. These extrinsic signals (<i>e.g.,</i> PDGF-AA, FGF-2, IGF-1, NT-3 or CNTF) may help to control the proper timing of OPC differentiation, ensuring adequate myelination and its maintenance. Surplus NG2 cells are eliminated by cell death in this balanced system.</p

PrP^c is expressed in oligodendrocyte precursor cells.

<p><b>A-C</b>) High magnification immunofluorescence images of double-labeled (NG2-PrP^c) cells in primary OPC cultures. <b>D</b>) PrP^c detection in Western blots of brain extracts from P0 <i>Prnp</i>^+/+ mice probed with the SAF61 antibody (lane 1) and extracts from primary postnatal oligodendrocyte cultures of <i>Prnp</i>^+/+ and <i>Prnp^0/0</i> mice (lanes 2 and 3 respectively). <b>E</b>) PrP^c detection in Western blots of embryonic mice ONs (E16.5) from <i>Prnp</i>^+/+ (lane 1) and <i>Prnp^0/0</i> mice (lane 2) probed with the SAF61 antibody. No PrP^c was detected in ONs from <i>Prnp^0/0</i> mice. <b>F-I</b>) Immunofluorescence images of <i>Prnp^0/0</i> (F-G) and <i>Prnp</i>^+/+ (H-I) SVZ neurospheres after differentiation, demonstrating the ability of both genotypes to differentiate into NG2-positive cells. No PrP^c expression was detected in NG2-positive cells in <i>Prnp^0/0</i> neurospheres. Scale bars: A  =  25 µm also applies to B-C; F  =  50 µm also applies to G-I.</p

The absence PrP^c increases OPC proliferation in cultured embryonic ONs.

<p><b>A</b>) Western blots of embryonic ONs (E16.5) from <i>Prnp^0/0</i> and <i>Prnp</i>^+/+ mice probed for Nestin and Olig2. Expression of both proteins was increased in <i>Prnp^0/0</i> versus <i>Prnp</i>^+/+ ONs. <b>B-C</b>) Representative immunofluorescence images of a 2 DIV ON explant culture from <i>Prnp</i>^+/+ mice in three dimensional collagen matrices after staining with anti-AB25 and counterstaining with DAPI. Cells that migrated radially were identified as OPCs by the expression of the A2B5 epitope (see also lower box). <b>D-G</b>) Immunofluorescence photomicrographs of 2 DIV ON explant cultures from <i>Prnp</i> (D-E) and <i>Prnp^0/0</i> (F-G) mice in three dimensional matrices after double staining with anti-BrdU and anti-A2B5. <b>H</b>) Scheme illustrating the method to quantify A2B5 and BrdU staining in explant cultures. Yellow boxes to quantify each quadrant are shown. <b>I</b>) Histogram showing relative percentage of A2B5 and BrdU-positive cells in <i>Prnp^0/0</i> and <i>Prnp^+/+</i> mice. The absence of PrP^c significantly increased the number of BrdU-labeled OPCs. <b>J</b>) Quantification of the maximal distance migrated by OPCs in ON explants. <i>Prnp</i>^+/+ OPCs migrated significantly further than their <i>Prnp^0/0</i> counterparts. Values in I and J represent the mean ± standard deviation, and the asterisks indicate statistical significance (<i>P</i> < 0.01, Student´s <i>t</i>-test). Scale bars: in B and D  =  200 µm also apply to A, and E-G, respectively.</p