Adenovirus-mediated transfection of caspase-8 sensitizes hepatocellular carcinoma to TRAIL- and chemotherapeutic agent-induced cell death

AbstractCaspase-8 belongs to the cysteine protease family and is known to be activated at the initial step in the cascade of TRAIL-induced apoptosis. The activation of procaspase-8 can be blocked by a relatively large amount of c-FLIP, which renders resistance to death receptor-mediated apoptosis in many types of cancer cells. To ask if extrinsic over-expression of caspase-8 contributes to the induction of apoptosis, we introduced the caspase-8 gene into HCC cells using an adenoviral (Adv) vector (Adv-Casp8). We demonstrated that Adv-Casp8 increased expression of active forms of caspase-8 in MOI-dependent manner. A large amount of Adv-Casp8 (MOI of 50) induced apoptosis significantly in HCC cells and resulted in downregulation of c-FLIP (in SK-Hep1, HLE, and HepG2 cells), XIAP, survivin, and Bcl-xL (in HLE cells) and dynamic release of cytochrome c and Smac from the mitochondria into the cytosol. On the other hand, a small amount of Adv-Casp8 (MOI of 10) causes a slight but detectable increase in the level of apoptosis with only a small effect on anti-apoptotic proteins and mitochondrial activation. However, small amounts of Adv-Casp8 augmented TRAIL- or chemotherapeutic agent-induced cell death (with an MOI of 10 or 20, respectively). These results suggest both that exogenous over-expression of caspase-8 by Adv-Casp8 may be essential for induction of HCC cell death and that the combination of Adv-Casp8 and TRAIL or chemotherapeutic agents could provide a useful strategy for treatment of HCC

Chemokines in bronchiolar epithelium in the development of chronic obstructive pulmonary disease.

The inflammatory chemokines interleukin-8, macrophage inflammatory protein-1, and monocyte chemoattractant protein-1, are reportedly involved in the pathogenesis of chronic obstructive pulmonary disease (COPD). Although bronchiolar epithelial cells and macrophages are known to be the cellular sources, the relative contribution of each cell type remains to be elucidated. In the present study, we first quantified cytokine mRNA in human bronchiolar epithelial cells and macrophages obtained using laser-capture microdissection and explored the relationship with early-stage COPD. Only in bronchiolar epithelial cells were interleukin-8, macrophage inflammatory protein-1, and monocyte chemoattractant protein-1 mRNA levels higher in smokers with airflow limitation and/or emphysema than those in never-smokers or smokers without either airflow limitation or emphysema. No difference was observed in macrophages. Complementary DNA (cDNA) array further revealed the overexpression of CC chemokine receptor 2 in bronchiolar epithelial cells from smokers with airflow limitation and/or emphysema. This study supports the role of bronchiolar epithelium as the source of increased inflammatory chemokine levels in the early development of COPD and also demonstrates the potential use of laser-capture microdissection, combined with reverse transcriptase–polymerase chain reaction and cDNA microarrays, to investigate functional profiles of individual structural and inflammatory cells in human lungs

COUPLING BETWEEN THE NH AND THE INTERMOLECULAR STRETCHING MODES OF 7-AZAINDOLE TAUTOMERIC DIMER

Author Institution: Department of Chemistry, Graduate School of Science, Kobe University, Nada-ku, Kobe 657-8501, JapanRecently, couplings between the intramolecular OH/NH and the intermolecular stretching modes of double hydrogen-bonded dimers, such as benzoic acid and 7-azaindole dimers, are investigated by the two-dimensional IR spectroscopy in solution. Infrared OH/NH stretching bands of such species exhibit a very complicated pattern. This pattern is analyzed theoretically based on the anharmonic coupling between the OH/NH stretching and the other modes. \par In the present study, we recorded infrared spectra of jet-cooled 7-AI tautomeric dimer by means of infrared-visible double resonance technique. This species is a product of the excited-state proton/hydrogen transfer reaction of 7-AI dimer and has the double hydrogen-bonded structure. Thus, the strong coupling between the intramolecular NH and the intermolecular stretching modes is expected as in the case of 7-AI normal dimer. In the IR spectrum observed in the present study, the NH stretching band exhibit a very broad width as expected. In addition, we have succeeded in recording the IR spectrum from the vibrational level involving an excitation of the intermolecular stretching mode. These results provide us with an effect of the coupling in the frequency-domain. The analysis of the vibrational coupling will be presented in the paper.\pa

Airflow limitation and airway dimensions in chronic obstructive pulmonary disease.

Rationale: Chronic obstructive pulmonary disease (COPD) is characterized by airflow limitation caused by emphysema and/or airway narrowing. Computed tomography has been widely used to assess emphysema severity, but less attention has been paid to the assessment of airway disease using computed tomography. Objectives: To obtain longitudinal images and accurately analyze short axis images of airways with an inner diameter 2 mm located anywhere in the lung with new software for measuring airway dimensions using curved multiplanar reconstruction. Methods: In 52 patients with clinically stable COPD (stage I, 14; stage II, 22; stage III, 14; stage IV, 2), we used the software to analyze the relationship of the airflow limitation index (FEV1, % predicted) with the airway dimensions from the third to the sixth generations of the apical bronchus (B1) of the right upper lobe and the anterior basal bronchus (B8) of the right lower lobe. Measurements and Main Results: Airway luminal area (Ai) and wall area percent (WA%) were significantly correlated with FEV1 (% predicted). More importantly, the correlation coefficients (r) improved as the airways became smaller in size from the third (segmental) to sixth generations in both bronchi (Ai: r = 0.26, 0.37, 0.58, and 0.64 for B1; r = 0.60, 0.65, 0.63, and 0.73 for B8). Conclusions: We are the first to use three-dimensional computed tomography to demonstrate that airflow limitation in COPD is more closely related to the dimensions of the distal (small) airways than proximal (large) airways