ABSTRACT
We evaluated the impact of selection of reference values on the categorisation of measured maximal oxygen consumption (V'O2peak) as "normal" or "abnormal" in an ageing population. We compared measured V'O2peak with predicted values and the lower limit of normal (LLN) calculated with five equations. 99 (58 males and 41 females) disease-free subjects aged ≥70â years completed an incremental maximal exercise test on a cycle ergometer. Mean V'O2peak was 1.88â L·min-1 in men and 1.26â L·min-1 in women. V'O2peak ranged from 89% to 108% of predicted in men, and from 88% to 164% of predicted in women, depending on the reference equation used. The proportion of subjects below the LLN ranged from 5% to 14% in men and 0-22% in women, depending on the reference equation. The LLN was lacking in one study, and was unsuitable for women in another. Most LLNs ranged between 53% and 73% of predicted. Therefore, choosing an 80% cut-off leads to overestimation of the proportion of "abnormal" subjects. To conclude, the proportion of subjects aged ≥70â years with a "low" V'O2peak differs markedly according to the chosen reference equations. In clinical practice, it is still relevant to test a sample of healthy volunteers and select the reference equations that better characterise this sample.
ABSTRACT
Chlamydia are obligate intracellular bacteria that cause variety of human diseases. Host cells infected with Chlamydia are protected against many different apoptotic stimuli. The induction of apoptosis resistance is thought to be an important immune escape mechanism allowing Chlamydia to replicate inside the host cell. Infection with C. trachomatis activates the Raf/MEK/ERK pathway and the PI3K/AKT pathway. Here we show that inhibition of these two pathways by chemical inhibitors sensitized C. trachomatis infected cells to granzyme B-mediated cell death. Infection leads to the Raf/MEK/ERK-mediated up-regulation and PI3K-dependent stabilization of the anti-apoptotic Bcl-2 family member Mcl-1. Consistently, interfering with Mcl-1 up-regulation sensitized infected cells for apoptosis induced via the TNF receptor, DNA damage, granzyme B and stress. Our data suggest that Mcl-1 up-regulation is primarily required to maintain apoptosis resistance in C. trachomatis-infected cells.
Subject(s)
Apoptosis , Chlamydia Infections/microbiology , Chlamydia trachomatis/metabolism , Gene Expression Regulation, Bacterial , Proto-Oncogene Proteins c-bcl-2/metabolism , Caspases/metabolism , Chlamydia Infections/metabolism , Enzyme Activation , Granzymes/pharmacology , HeLa Cells , Humans , MAP Kinase Signaling System , Models, Biological , Myeloid Cell Leukemia Sequence 1 Protein , Phosphatidylinositol 3-Kinases/metabolism , RNA, Small Interfering/metabolismABSTRACT
Mitochondria play a pivotal role during stress-induced apoptosis as several proapoptotic proteins are released to the cytosol to activate caspases. Smac/DIABLO is one of the proapoptotic proteins released from the mitochondria and has been shown to inactivate IAPs. However, gene knockout studies in mice revealed a redundant role for Smac during development and cell death. By applying RNA interference-mediated loss of function approach, we demonstrate that Smac/DIABLO is required for the activation of effector but not initiator caspases during stress and receptor-mediated cell death in HeLa cells. Cells with reduced Smac resist apoptosis and retained clonogenicity. Our results suggest an obligatory role for Smac/DIABLO in these tumor cells during several pathways of apoptosis induction.
Subject(s)
Apoptosis , Caspases, Effector/metabolism , Intracellular Signaling Peptides and Proteins/physiology , Mitochondrial Proteins/physiology , Apoptosis/genetics , Apoptosis Regulatory Proteins , Brefeldin A/pharmacology , Enzyme Activation/genetics , HeLa Cells , Humans , Intracellular Signaling Peptides and Proteins/genetics , Mitochondrial Proteins/genetics , Protein Synthesis Inhibitors/pharmacology , RNA Interference , TransfectionABSTRACT
Host cells infected with obligate intracellular bacteria Chlamydia trachomatis are profoundly resistant to diverse apoptotic stimuli. The molecular mechanisms underlying the block in apoptotic signaling of infected cells is not well understood. Here we investigated the molecular mechanism by which apoptosis induced via the tumor necrosis factor (TNF) receptor is prevented in infected epithelial cells. Infection with C. trachomatis leads to the up-regulation of cellular inhibitor of apoptosis (cIAP)-2, and interfering with cIAP-2 up-regulation sensitized infected cells for TNF-induced apoptosis. Interestingly, besides cIAP-2, cIAP-1 and X-linked IAP, although not differentially regulated by infection, are required to maintain apoptosis resistance in infected cells. We detected that IAPs are constitutively organized in heteromeric complexes and small interfering RNA-mediated silencing of one of these IAPs affects the stability of another IAP. In particular, the stability of cIAP-2 is modulated by the presence of X-linked IAP and their interaction is stabilized in infected cells. Our observations suggest that IAPs are functional and stable as heteromers, a thus far undiscovered mechanism of IAP regulation and its role in modulation of apoptosis.
