Journal: Journal of Biomedicine and Biotechnology

Article Title: Oxaprozin-Induced Apoptosis on CD40 Ligand-Treated Human Primary Monocytes Is Associated with the Modulation of Defined Intracellular Pathways

doi: 10.1155/2009/478785

Figure Lengend Snippet: Oxaprozin increases apoptosis in CD40L-treated monocytes . Monocytes were incubated for 48 hours with control medium or 200 ng/mL CD40L plus 1 μ g/mL CD40L enhancer in the presence or absence of different doses (0, 5, 10, 50,100 μ M) of oxaprozin. (a) Assessment of apoptosis by acridine-orange-stained slides under fluorescence microscopy. Data are express as percentages of apoptotic cells on total number of cells counted (mean ± SEM, n = 3; apoptosis in absence versus in presence of CD40L: * P < .05; apoptosis in presence of CD40L versus CD40L plus 10 or 50 μ M oxaprozin: # P < .05; apoptosis in presence of CD40L versus CD40L plus 100 μ M oxaprozin: ## P < .01). (b) Assessment of apoptosis by using Annexin V and Propidium Iodide (PI) binding assay. (mean ± SEM, n = 4; apoptosis in absence versus in presence of CD40L: * P < .05; apoptosis in presence of CD40L versus CD40L plus 100 μ M oxaprozin: # P < .05 and ## P < .01).

Article Snippet: In selective experiments, cells were cultured in the presence or absence of 50 μ M PD98059 (MEK inhibitor, a kinase activating ERK 1/2, from Biomol Research Laboratories, Inc., Plymouth Meeting, PA), 1 μ M SB203580 (p38 MAPK inhibitor, from Biomol Research Laboratories, Inc.), 50 μ M LY294002 (PI3K inhibitor, from Sigma-Aldrich), 20 μ M SN-50 (NF- κ B inhibitor, from Calbiochem, San Diego, CA), 50 μ M Ac-DEVD-CHO (caspase 3 inhibitor, from Bachem AG, Bubendorf, Switzerland), different doses (5, 10, 50, 100 μ M) of oxaprozin (Helsinn Healthcare SA, Lugano, Switzerland), 100 μ M ibuprofen (Sigma-Aldrich),100 μ M indomethacin (Sigma-Aldrich), or 100 μ M naproxene (Sigma-Aldrich).

Techniques: Incubation, Staining, Fluorescence, Microscopy, Binding Assay

Journal: Journal of Biomedicine and Biotechnology

Article Title: Oxaprozin-Induced Apoptosis on CD40 Ligand-Treated Human Primary Monocytes Is Associated with the Modulation of Defined Intracellular Pathways

doi: 10.1155/2009/478785

Figure Lengend Snippet: Oxaprozin-induced apoptosis is independent on COX inhibition . Monocytes were incubated for 48 hours with control medium or 200 ng/mL CD40L plus 1 μ g/mL CD40L enhancer in the presence or absence of 100 μ M oxaprozin, 100 μ M ibuprofen, 100 μ M indomethacin, or 100 μ M naproxene. (a) Assessment of apoptosis by acridine-orange-stained slides under fluorescence microscopy. Data are express, as percentages of apoptotic cells on total number of cells counted (mean ± SEM, n = 4; apoptosis in absence versus in presence of CD40L: * P < .05; apoptosis in presence of CD40L versus CD40L plus oxaprozin: # P < .05; apoptosis in presence of CD40L versus CD40L plus other NSAIDs: N.S.). (b) Assessment of apoptosis by using Annexin V and Propidium Iodide (PI) binding assay (mean ± SEM, n = 3; apoptosis in absence versus in presence of CD40L: * P < .05; apoptosis in presence of CD40L versus CD40L plus oxaprozin: # P < .05; apoptosis in presence of CD40L versus CD40L plus other NSAIDs: N.S.).

Article Snippet: In selective experiments, cells were cultured in the presence or absence of 50 μ M PD98059 (MEK inhibitor, a kinase activating ERK 1/2, from Biomol Research Laboratories, Inc., Plymouth Meeting, PA), 1 μ M SB203580 (p38 MAPK inhibitor, from Biomol Research Laboratories, Inc.), 50 μ M LY294002 (PI3K inhibitor, from Sigma-Aldrich), 20 μ M SN-50 (NF- κ B inhibitor, from Calbiochem, San Diego, CA), 50 μ M Ac-DEVD-CHO (caspase 3 inhibitor, from Bachem AG, Bubendorf, Switzerland), different doses (5, 10, 50, 100 μ M) of oxaprozin (Helsinn Healthcare SA, Lugano, Switzerland), 100 μ M ibuprofen (Sigma-Aldrich),100 μ M indomethacin (Sigma-Aldrich), or 100 μ M naproxene (Sigma-Aldrich).

Techniques: Inhibition, Incubation, Staining, Fluorescence, Microscopy, Binding Assay

Journal: Journal of Biomedicine and Biotechnology

Article Title: Oxaprozin-Induced Apoptosis on CD40 Ligand-Treated Human Primary Monocytes Is Associated with the Modulation of Defined Intracellular Pathways

doi: 10.1155/2009/478785

Figure Lengend Snippet: Oxaprozin-induced apoptosis involves the inhibition of CD40L-induced Akt and N F - κ Bphosphorylation . Monocytes were incubated for 48 hours with control medium or 200 ng/mL CD40L plus 1 μ g/mL CD40L enhancer in the presence or absence of 50 μ M LY294002 (LY), 50 μ M PD98059 (PD), 1 μ M SB203580 (SB), or 20 μ M SN-50. (a) Assessment of apoptosis on cytopreps stained with acridine orange. Data are expressed as percentages of apoptotic cells on total number of cells counted (mean ± SEM, n = 3, apoptosis in absence versus in presence of CD40L: ** P < .01; apoptosis in presence of CD40L versus CD40L plus PD: ## P < .01; apoptosis in presence of CD40L versus CD40L plus LY or SB or SN-50: # P < .05). (b) Assessment of apoptosis using Annexin V and Propidium Iodide (PI) binding assay. Data are expressed as percentages of positive cells on total number of cells counted (mean ± SEM, n = 2). (c) Monocytes were prencubated in the presence or absence of 100 μ M oxaprozin for 1 hour and then stimulated with control medium or 200 ng/mL CD40L plus 1 μ g/mL CD40L enhancer for 15 minutes. Then, cells were lysed, and western blot analysis was performed. Results represent one of three experiments that yielded similar results.

Article Snippet: In selective experiments, cells were cultured in the presence or absence of 50 μ M PD98059 (MEK inhibitor, a kinase activating ERK 1/2, from Biomol Research Laboratories, Inc., Plymouth Meeting, PA), 1 μ M SB203580 (p38 MAPK inhibitor, from Biomol Research Laboratories, Inc.), 50 μ M LY294002 (PI3K inhibitor, from Sigma-Aldrich), 20 μ M SN-50 (NF- κ B inhibitor, from Calbiochem, San Diego, CA), 50 μ M Ac-DEVD-CHO (caspase 3 inhibitor, from Bachem AG, Bubendorf, Switzerland), different doses (5, 10, 50, 100 μ M) of oxaprozin (Helsinn Healthcare SA, Lugano, Switzerland), 100 μ M ibuprofen (Sigma-Aldrich),100 μ M indomethacin (Sigma-Aldrich), or 100 μ M naproxene (Sigma-Aldrich).

Techniques: Inhibition, Incubation, Staining, Binding Assay, Western Blot

Journal: Journal of Biomedicine and Biotechnology

Article Title: Oxaprozin-Induced Apoptosis on CD40 Ligand-Treated Human Primary Monocytes Is Associated with the Modulation of Defined Intracellular Pathways

doi: 10.1155/2009/478785

Figure Lengend Snippet: Oxaprozin-induced apoptosis of CD40L-treated monocytes involves the activation of caspase 3 . (a) Monocytes were cultured for 48 hours with control medium or 200 ng/mL CD40L plus 1 μ g/mL CD40L enhancer in the presence or absence of 100 μ M of oxaprozin. Then cells were lysed, and caspase 3 activity was determined by spectrophotometer. Data are expressed as mean ± SEM, n = 3. (caspase 3 activity in absence versus in presence of CD40L: * P < .05; caspase 3 activity in presence of CD40L versus CD40L plus oxaprozin: # P < .05). (b) Monocytes were incubated for 48 hours with control medium or 200 ng/mL CD40L plus 1 μ g/mL CD40L enhancer in the presence or absence 100 μ M of oxaprozin and 50 μ M Ac-DEVD-CHO (caspase 3 inhibitor, CASP 3 INHIB). Data are expressed as mean ± SEM, n = 3 (apoptosis in absence versus in presence of CD40L: * P < .05; apoptosis in presence of CD40L versus CD40L plus oxaprozin: # P < .05; apoptosis in presence of CD40L plus oxaprozin versus CD40L plus oxaprozin and caspase 3 inhibitor: § P < .05).

Article Snippet: In selective experiments, cells were cultured in the presence or absence of 50 μ M PD98059 (MEK inhibitor, a kinase activating ERK 1/2, from Biomol Research Laboratories, Inc., Plymouth Meeting, PA), 1 μ M SB203580 (p38 MAPK inhibitor, from Biomol Research Laboratories, Inc.), 50 μ M LY294002 (PI3K inhibitor, from Sigma-Aldrich), 20 μ M SN-50 (NF- κ B inhibitor, from Calbiochem, San Diego, CA), 50 μ M Ac-DEVD-CHO (caspase 3 inhibitor, from Bachem AG, Bubendorf, Switzerland), different doses (5, 10, 50, 100 μ M) of oxaprozin (Helsinn Healthcare SA, Lugano, Switzerland), 100 μ M ibuprofen (Sigma-Aldrich),100 μ M indomethacin (Sigma-Aldrich), or 100 μ M naproxene (Sigma-Aldrich).

Techniques: Activation Assay, Cell Culture, Activity Assay, Spectrophotometry, Incubation, Inhibition

Journal: Journal of Biomedicine and Biotechnology

Article Title: Oxaprozin-Induced Apoptosis on CD40 Ligand-Treated Human Primary Monocytes Is Associated with the Modulation of Defined Intracellular Pathways

doi: 10.1155/2009/478785

Figure Lengend Snippet: (a) CD40L-induced monocyte survival is associated with Akt, ERK 1/2, P38 MAPK and NF- κ B activation, and caspase 3 inhibition. (b) Oxaprozin-induced apoptosis in CD40L-treated monocytes is associated with the inhibition of Akt and NF- κ B and the activation of caspase 3 pathways.

Article Snippet: In selective experiments, cells were cultured in the presence or absence of 50 μ M PD98059 (MEK inhibitor, a kinase activating ERK 1/2, from Biomol Research Laboratories, Inc., Plymouth Meeting, PA), 1 μ M SB203580 (p38 MAPK inhibitor, from Biomol Research Laboratories, Inc.), 50 μ M LY294002 (PI3K inhibitor, from Sigma-Aldrich), 20 μ M SN-50 (NF- κ B inhibitor, from Calbiochem, San Diego, CA), 50 μ M Ac-DEVD-CHO (caspase 3 inhibitor, from Bachem AG, Bubendorf, Switzerland), different doses (5, 10, 50, 100 μ M) of oxaprozin (Helsinn Healthcare SA, Lugano, Switzerland), 100 μ M ibuprofen (Sigma-Aldrich),100 μ M indomethacin (Sigma-Aldrich), or 100 μ M naproxene (Sigma-Aldrich).

Techniques: Activation Assay, Inhibition