The present study explored the role of the cell surface receptor Fas (CD95/APO-1) in apoptosis induced by camptothecin (CPT) in the HT29 colon carcinoma cell line. CPT-induced apoptosis was associated with high molecular weight DNA fragmentation as measured by ®lter elution. This fragmentation was inhibited by the caspase inhibitor, z-VAD-fmk and by cycloheximide, which also prevented proteolytic activation of caspase-3 and poly(ADP-ribose)polymerase cleavage. Under such conditions, Fas, Fas ligand, Bax, and p21 expression were increased and Fas recruited the FADD adaptor. Fas expression increase was blocked by cycloheximide but not by z-VAD-fmk, consistent with caspase activation downstream from Fas. Treatment of HT29 cells with FasL or with the CH-11 agonistic anti-Fas antibody potentiated the apoptotic response of cells treated with CPT. The anti-Fas blocking antibody ZB4 and the Fasligand inhibitor failed to protect HT29 cells from CPTinduced apoptosis. Such a protection was obtained by transient expression of constructs encoding a dominantnegative mutant of FADD, FADD in an antisense orientation and E8 or MC159 viral proteins that inhibit Fas-induced apoptosis at the level of FADD and procaspase-8, respectively. Together, these data show that topoisomerase I-mediated DNA damage-induced apoptosis involves activation of the Fas pathway without detectable Fas-ligand requirement in CPT-treated cells.

Objectives: To investigate the predictive value of four-dimensional (4D) strain echocardiography for major adverse cardiovascular events (MACE) in ST-elevation acute myocardial infarction (STEMI) patients. Methods: Consecutive STEMI patients who underwent successful primary coronary interven tion (PCI) were enrolled and followed, with 2D and 4D strain echocardiography performed within 1 week after PCI. Results: Twenty-six first MACE were recorded in 81 patients who finished a ∼3.0 year follow-up. Compared with those without MACE, subjects with MACE were more likely to have anterior MI (73.08 vs. 38.18%, p = 0.003), significantly decreased 2D left ventricular ejection fraction (2DLVEF) and 4DLVEF (all p < 0.05), as well as an overtly compromised 4D strain parameters. The prediction models incorporating infarct location with either 2DLVEF or 4D strain parameters were then developed. Model comparisons revealed that the global area strain (GAS)-based model had the highest discriminative capacity (c statistics = 0.774) and was well calibrated for MACE. Additionally, the clinical utility of the GAS-based prediction model was verified by decision curve analysis showing a consistent positive and larger net benefit compared to the 2DLVEF-based model. Conclusions: Our data support a superiority of 4D strain echocardiography over conventional 2D echocardiography, especially GAS, for risk stratification in STEMI patients after successful primary PCI.

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Aim: The present study examined the differential expression of proteins in HuH‐7 cells and HuH‐7 cells harboring in vitro‐transcribed full‐length hepatitis C virus 1b RNA (HuH‐7‐HCV), and elucidated the cellular responses to HCV replication. Methods: The protein profiles of matched pairs of HuH‐7‐HCV cells and HuH‐7 mock cells were analyzed by 2‐D electrophoresis (2DE). Solubilized proteins were separated in the first dimension by isoelectric focusing, and by 12.5% SDS‐PAGE in the second dimension. The differential protein expression was analyzed by use of image analysis software to identify candidates for HCV infection‐associated proteins. Results: In total, 29 protein spots showed increases and 25 protein spots showed decreases in signal in HuH‐7‐HCV cell 2DE profiles as compared with HuH‐7 mock cells. In the next step, the 10 spots showing the greatest increase and the 10 spots showing the greatest decrease were excised from gels and the proteins present were identified by Matrix‐Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometer (MALDI‐TOF MS) or MALDI‐TOF/TOF MS. In total, 13 proteins were identified successfully. The potential significance of the differential expression due to HCV replication was discussed. Conclusion: Our study identifies changes in the proteome of HuH‐7 cells in the presence of HCV replication and yields information of the mechanism of HCV pathogenesis. These results will be useful for the identification of HCV infection‐associated proteins that could be molecular targets for treatment.