Cells were incubated with chloroquine for 2 hours before stimulation with lipopolysaccharide (LPS, 1g/ml). Cell viability was measured by staining cells with 3-(4,5-dimethylthiazol-2- yl)-2,5- diphenyltetrazolium bromide. TNF in the culture supernatant and the cell lysate were measured by sandwich ELISA, using specific monoclonal antibodies according to the manufacturer's instructions. To determine the level of cell-associated cytokines, cells were washed with phosphate-buffered saline (PBS) and lysed in a buffer containing 0.5% NP-40. Standard recombinant proteins were also diluted in PBS containing the same concentrations of NP-40 as the cell lysate. Cytokine synthesized were proteolytically cleaved and released into medium. The amount of cytokines in the culture supernatant and the cell lysate were presented. Results for TNF production are shown in Figure 1 (Jang et al, 2006). (Cells: PBMC: Human peripheral blood mononuclear cells, monocytes, monocytic lymphoma U-937 and leukemia THP1 cells). TNF (ng) 30 20 10 0 15 10 5 0 1.5 LL H PBMCs Medium Cell Monocytes U937

Cells were incubated with chloroquine for 2 hours before stimulation with lipopolysaccharide (LPS, 1g/ml). Cell viability was measured by staining cells with 3-(4,5-dimethylthiazol-2- yl)-2,5- diphenyltetrazolium bromide. TNF in the culture supernatant and the cell lysate were measured by sandwich ELISA, using specific monoclonal antibodies according to the manufacturer's instructions. To determine the level of cell-associated cytokines, cells were washed with phosphate-buffered saline (PBS) and lysed in a buffer containing 0.5% NP-40. Standard recombinant proteins were also diluted in PBS containing the same concentrations of NP-40 as the cell lysate. Cytokine synthesized were proteolytically cleaved and released into medium. The amount of cytokines in the culture supernatant and the cell lysate were presented. Results for TNF production are shown in Figure 1 (Jang et al, 2006). (Cells: PBMC: Human peripheral blood mononuclear cells, monocytes, monocytic lymphoma U-937 and leukemia THP1 cells). TNF (ng) 30 20 10 0 15 10 5 0 1.5 LL H PBMCs Medium Cell Monocytes U937

Curren'S Math For Meds: Dosages & Sol

11th Edition

ISBN:9781305143531

Author:CURREN

Publisher:CURREN

Chapter16: Adult And Pediatric Dosages Based On Body Surface Area

Section: Chapter Questions

Problem 7.1P

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