Ascites are commonly found in ovarian cancer patients with advanced disease and are rich in cellular components and growthpromoting factors. The purpose of this study was to assess the effect of malignant ascites on TRAIL-induced apoptosis. We demonstrate that malignant ascites obtained from women with advanced ovarian cancer protect tumor cells from TRAIL-and FasL-induced apoptosis but not against cisplatin-induced apoptosis. This antiapoptotic effect was consistently found among different malignant ascites while nonmalignant peritoneal fluids or conditioned medium from TRAIL-resistant cells failed to protect tumor cells against TRAIL killing. Malignant ascites strongly inhibits TRAIL-induced caspase-3 activation and PARP cleavage. Furthermore, ascites activate PI3K and its downstream target Akt and increases c-FLIP S protein levels without affecting ERK phosphorylation status. The antiapoptotic effect of malignant ascites is abrogated by the inhibition of PI3K with LY294002, by a specific inhibitor of Akt and by Akt siRNA. We further show that the prosurvival effect of ascites can be suppressed by down-regulation of c-FLIP S . Our data indicate that malignant effusions protect against TRAIL-induced apoptosis by activating the PI3K/Akt pathway. These findings demonstrate that the tumor microenvironment may contribute to the resistance of ovarian cancer cells to death receptor-induced apoptosis. ' 2007 Wiley-Liss, Inc.

Little is known on how cancer cells can acquire resistance to tumor necrosis factor -related apoptosis-inducing ligand (TRAIL). In this study, we established TRAIL-resistant cells from the TRAIL-sensitive human ovarian carcinoma cell line OVCAR3 to evaluate the potential mechanisms of acquired resistance to TRAIL. The selected resistant cells were cross-resistant to Fas ligand but remained sensitive to drug-induced apoptosis. Expression of TRAIL receptors was not altered in TRAIL-resistant OVCAR3 cells. Cleavage of caspase-8 and caspase-3 occurred in both TRAILresistant and TRAIL-sensitive cells. However, mature caspase-3 fragments were not detected by immunoblot in TRAIL-resistant cells and caspase-3 activity was significantly inhibited in these cells. The addition of proteasome inhibitors significantly increased TRAIL-induced apoptosis in resistant cells and enhanced the accumulation of mature caspase-3 fragments. Pretreatment with cycloheximide showed that active caspase-3 fragments have a high turnover rate in OVCAR3 R350 cells. X-linked inhibitor of apoptosis down-regulation by RNA interference also increased the accumulation of cleaved caspase-3 intermediates and resensitized TRAIL-resistant cells. Our findings show that altered turnover of mature caspase-3 may lead to acquired TRAIL resistance in ovarian cancer cells. Proteasome and X-linked inhibitor of apoptosis inhibitors could have a role in clinical situations to potentiate the cytotoxic effects of TRAIL in resistant tumor cells. [Mol Cancer Ther 2006;5(3):509 -21]

We evaluated the effect of heat challenge on cell viability, concanavalin A-induced proliferation and heat shock protein (HSPs) mRNA expression in peripheral mononuclear blood cells (PBMC) isolated from Creole (CR) and Large White (LW) pigs. The PBMCs were cultured for 9 h at 37 °C before being subjected to heat challenge: (1) at 42 °C or 45 °C for 2, 4, 6 and 9 h to monitor cell viability;(2) at 45 °C for 2 and 9 h followed by stimulation for 24 h at 37 °C with concanavalin A to evaluate mitogen-induced proliferation; and (3) at 45 °C for 3, 6 and 9 h to measure induction of HSP70.2 and HSP90 mRNA. Cell viability was affected by breed and temperature (P < 0.01), and the viability decrease caused by heat challenge was greater for LW than CR pigs. For mitogen-stimulated PBMCs, incubation at 45 °C reduced lymphoblastogenesis equally in both breeds (P < 0.01). Although heat challenge for 3 and 6 h at 45°C induced expression of HSP70.2 and HSP90 mRNA, no breed difference was observed. In conclusion, differences in heat resistance between these two breeds at the whole organism level are reflected at the cellular level. Neither HSP70.2 nor HSP90 mRNA expression levels explain this effect.

The cessation of driving is a difficult transition for the elderly, but it can be facilitated through interventions. The purpose of this study was to explore the satisfaction, usefulness and applicability of the CarFreeMe intervention in the French-Canadian context. A qualitative clinical research device was used on ten older adults aged between 61 and 90 years. The participants had stopped driving within the last twelve months or were planning to stop driving in the near future and did not have cognitive impairments. After the intervention, the participants were generally satisfied and reported on its usefulness and applicability in a French-Canadian context. In addition, they identified the positive impacts related to their social involvement as they re-engaged in or pursued their significant activities. Further research is required to assess the intervention’s effects and the practicability of implementing it in Canada.

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