Journal: Environmental Health Perspectives

Article Title: Seasonal Variations in Air Pollution Particle-Induced Inflammatory Mediator Release and Oxidative Stress

doi: 10.1289/ehp.7996

Figure Lengend Snippet: Production of ( A ) IL-6 by AM and ( B ) IL-8 by NHBE cells stimulated with coarse, fine, and ultrafine Chapel Hill pollution particles collected in four different months. The control IL-8 and IL-6 concentrations were 1.7 ± 0.2 ng/mL and 198 ± 5 pg/mL, respectively. Particles were added to NHBE cells at 11 μg/mL and to AMs at 50 μg/mL. *Tukey adjusted p -value < 0.05; n = 3–4 each. The dashed line denotes 1.0 (no change over control). The brackets indicate the groups that are significantly different by ANOVA and the Tukey subtest.

Article Snippet: Typically at confluency, NHBE cells in the 12-well plate (catalog no. 3512; Costar, Corning, NY) had a density of 300,000–500,000 cells/well in 2 mL of BEGM.

Techniques:

Journal: Environmental Health Perspectives

Article Title: Seasonal Variations in Air Pollution Particle-Induced Inflammatory Mediator Release and Oxidative Stress

doi: 10.1289/ehp.7996

Figure Lengend Snippet: Production of ROSs measured by DCF in ( A ) AM and ( B ) NHBE cells stimulated with coarse, fine, and ultrafine Chapel Hill pollution particles collected in four different months. Particles were added to NHBE cells at 11 μg/mL and to AMs at 50 μg/mL. In general, NHBE cells appeared to be more responsive to PM than the AMs because increases in DCF signals were similar between the two cell types even though NHBE cells were exposed to lower doses of PM. *Tukey adjusted p -value < 0.05; # p < 0.05 versus control (ratio = 1.0) by the one-group Student t -test; n = 3–4 each. The dashed line denotes 1.0 (no change over control). The brackets indicate groups that are significantly different by the Tukey subtest.

Article Snippet: Typically at confluency, NHBE cells in the 12-well plate (catalog no. 3512; Costar, Corning, NY) had a density of 300,000–500,000 cells/well in 2 mL of BEGM.

Techniques:

Journal: Environmental Health Perspectives

Article Title: Seasonal Variations in Air Pollution Particle-Induced Inflammatory Mediator Release and Oxidative Stress

doi: 10.1289/ehp.7996

Figure Lengend Snippet: Production of ROSs measured by DHR in ( A ) AM and ( B ) NHBE cells stimulated with coarse, fine, and ultrafine Chapel Hill pollution particles collected in four different months. Particles were added to NHBE cells at 11 μg/mL and to AMs at 50 μg/mL. In general, NHBE cells appeared more responsive to PM than were AMs because increases in DHR signals were similar between the two cell types even though NHBE cells were exposed to lower doses of PM. *Tukey adjusted p -value < 0.05; n = 3–4 each. The dashed line denotes 1.0 (no change over control). The brackets indicate groups that are significantly different by ANOVA and the Tukey subtest.

Article Snippet: Typically at confluency, NHBE cells in the 12-well plate (catalog no. 3512; Costar, Corning, NY) had a density of 300,000–500,000 cells/well in 2 mL of BEGM.

Techniques:

Journal: Environmental Health Perspectives

Article Title: Seasonal Variations in Air Pollution Particle-Induced Inflammatory Mediator Release and Oxidative Stress

doi: 10.1289/ehp.7996

Figure Lengend Snippet: Effects of PM from 12 different months on the release of ( A ) IL-6 in AMs and ( B ) IL-8 in NHBE cells. Particles were added to AMs at 50 μg/mL and to NHBE cells at 11 μg/mL. The data for December ultrafine PM were not available and were omitted in the figure; n = 3–4 each.

Article Snippet: Typically at confluency, NHBE cells in the 12-well plate (catalog no. 3512; Costar, Corning, NY) had a density of 300,000–500,000 cells/well in 2 mL of BEGM.

Techniques:

Journal: Environmental Health Perspectives

Article Title: Seasonal Variations in Air Pollution Particle-Induced Inflammatory Mediator Release and Oxidative Stress

doi: 10.1289/ehp.7996

Figure Lengend Snippet: Correlations between Cr and IL-8 release in NHBE cells incubated with ( A ) coarse, ( B ) fine, and ( C ) ultrafine PM from 12 months. The dashed line represents the linear regression line. The overall model p -values for coarse, fine, and ultrafine PM were 0.106, 0.003, and 0.036, respectively. R 2 = 0.6082 and 0.401 for fine and ultrafine PM, respectively.

Article Snippet: Typically at confluency, NHBE cells in the 12-well plate (catalog no. 3512; Costar, Corning, NY) had a density of 300,000–500,000 cells/well in 2 mL of BEGM.

Techniques: Incubation

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Mechanistic Contribution of Ubiquitous 15-Lipoxygenase-1 Expression Loss in Cancer Cells to Terminal Cell Differentiation Evasion

doi: 10.1158/1940-6207.CAPR-10-0280

Figure Lengend Snippet: 15-LOX-1 and differentiation of primary NHBE cells. Primary NHBE cells were grown for 3 weeks in an undifferentiated state in immersion cultures or in air-liquid interface cultures to induce terminal differentiation into bronchial epithelial-like structures. Cells were then harvested and processed for quantitative real-time reverse transcription-polymerase chain reaction (panel A, differentiated vs. undifferentiated NHBE cells, P = 0.0005), Western blotting (panel B), and 13-HODE levels by liquid chromatography tandem mass spectroscopy (panel C, differentiated vs. undifferentiated NHBE cells, P = 0.0002). + represents 15-LOX-1 positive control (HCT-116 colon cancer cells transfected with 15-LOX-1 expression vector).

Article Snippet: We obtained from MatTek Corporation (Ashland, MA) primary NHBE cells (originally provided by Clonetics [San Diego, CA)]) grown in an in vitro system to form a pseudostratified, highly differentiated mucociliary epithelium that closely resembles the epithelial tissue of the respiratory tract (differentiated NHBE cells) ( Supplementary Fig. S1A ) and NHBE cells grown in an undifferentiated state in submerged (immersion) cultures (undifferentiated NHBE cells) ( Supplementary Fig. S1B ) ( 22 , 23 ).

Techniques: Reverse Transcription, Polymerase Chain Reaction, Western Blot, Liquid Chromatography, Tandem Mass Spectroscopy, Positive Control, Transfection, Expressing, Plasmid Preparation

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Mechanistic Contribution of Ubiquitous 15-Lipoxygenase-1 Expression Loss in Cancer Cells to Terminal Cell Differentiation Evasion

doi: 10.1158/1940-6207.CAPR-10-0280

Figure Lengend Snippet: 15-LOX-1 and P16 mRNA and protein expression levels in cancer cell lines. A and B, A total of 128 cancer cell lines (Supplementary Table S1) were cultured and processed for 15-LOX-1 (panel A) and p16 (panel B) mRNA by quantitative real-time reverse transcription-polymerase chain reaction. Dots in the dot plots are means of triplicate measurements from each cell line. The relative expression levels were calculated relative to expression of the calibrator sample (differentiated NHBE cells). Solid lines represent the median value for each group. C, 15-LOX-1 relative expression levels in cancer cell lines compared to the level in Caco-2 cells terminally differentiated by sodium butyrate treatment. 15-LOX-1 mRNA measurements are as in panel A but with terminally differentiated Caco-2 cells as the calibrator sample. Dots in the dot plots are means of triplicate measurements from each cell line. The solid line represents the median value for the relative expression levels. D, 15-LOX-1 protein expression in cancer cell lines. Cell lines—including cell lines with 15-LOX-1 mRNA expression levels nearly equal to or greater than the level in differentiated Caco-2 cells or NHEK—were cultured, processed for Western blotting, and probed with 15-LOX-1 antibody. Three repeated experiments yielded similar results.

Article Snippet: We obtained from MatTek Corporation (Ashland, MA) primary NHBE cells (originally provided by Clonetics [San Diego, CA)]) grown in an in vitro system to form a pseudostratified, highly differentiated mucociliary epithelium that closely resembles the epithelial tissue of the respiratory tract (differentiated NHBE cells) ( Supplementary Fig. S1A ) and NHBE cells grown in an undifferentiated state in submerged (immersion) cultures (undifferentiated NHBE cells) ( Supplementary Fig. S1B ) ( 22 , 23 ).

Techniques: Expressing, Cell Culture, Reverse Transcription, Polymerase Chain Reaction, Western Blot