The study objective was to establish The American Association for Thoracic Surgery (AATS) evidence-based guidelines for the management of empyema.

dWhooping cough results from infection of the respiratory tract with Bordetella pertussis, and the secreted adenylate cyclase toxin (ACT) is essential for the bacterium to establish infection. Despite extensive study of the mechanism of ACT cytotoxicity and its effects over a range of concentrations in vitro, ACT has not been observed or quantified in vivo, and thus the concentration of ACT at the site of infection is unknown. The recently developed baboon model of infection mimics the prolonged cough and transmissibility of pertussis, and we hypothesized that measurement of ACT in nasopharyngeal washes (NPW) from baboons, combined with human and in vitro data, would provide an estimate of the ACT concentration in the airway during infection. NPW contained up to ϳ10 8 CFU/ml B. pertussis and 1 to 5 ng/ml ACT at the peak of infection. Nasal aspirate specimens from two human infants with pertussis contained bacterial concentrations similar to those in the baboons, with 12 to 20 ng/ml ACT. When ϳ10 8 CFU/ml of a laboratory strain of B. pertussis was cultured in vitro, ACT production was detected in 60 min and reached a plateau of ϳ60 ng/ml in 6 h. Furthermore, when bacteria were brought into close proximity to target cells by centrifugation, intoxication was increased 4-fold. Collectively, these data suggest that at the bacterium-target cell interface during infection of the respiratory tract, the concentration of ACT can exceed 100 ng/ml, providing a reference point for future studies of ACT and pertussis pathogenesis.

The adenylate cyclase toxin (ACT) of Bordetella pertussis intoxicates target cells by generating supraphysiologic levels of intracellular cyclic AMP (cAMP). Since ACT kills macrophages rapidly and potently, we asked whether ACT would also kill neutrophils. In fact, ACT prolongs the neutrophil life span by inhibiting constitutive apoptosis and preventing apoptosis induced by exposure to live B. pertussis. Imaging of B. pertussis-exposed neutrophils revealed that B. pertussis lacking ACT induces formation of neutrophil extracellular traps (NETs), whereas wild-type B. pertussis does not, suggesting that ACT suppresses NET formation. Indeed, ACT inhibits formation of NETs by generating cAMP and consequently inhibiting the oxidative burst. Convalescent-phase serum from humans following clinical pertussis blocks the ACT-mediated suppression of NET formation. These studies provide novel insight into the phagocyte impotence caused by ACT, which not only impairs neutrophil function but also inhibits death of neutrophils by apoptosis and NETosis.

Ciliary neurotrophic factor (CNTF)-dependent induction of expression of the neuropeptide vasoactive intestinal peptide (VIP) gene is mediated by a 180-base pair cytokine response element (CyRE) in the VIP promoter.To elucidate the molecular mechanisms mediating the transcriptional activation by CNTF, intracellular signaling to the CyRE has been studied in a neuroblastoma cell line. It has been shown previously that CNTF induces Stat proteins to bind to a site within the CyRE. CNTF also induces a second protein to bind to a C/EBPlike site within the CyRE. In this report, we show that this inducible CyRE binding protein is composed of the AP-1 proteins c-Fos, JunB, and JunD. These proteins bind to a non-canonical AP-1 site located near the previously characterized C/EBP site. The serine/threonine kinase inhibitor H7 prevents CNTF-dependent induction of AP-1 binding and CyRE-mediated transcription, suggesting that an H7-sensitive kinase is important to mediating CNTF effects on VIP transcription. The integration at the VIP CyRE of the Jak-Stat and AP-1 signaling pathways with other pre-existing proteins provides a cellular mechanism for cell-and cytokine-specific signaling.Ciliary neurotrophic factor (CNTF) 1 is a member of the neuropoietic cytokine family, which includes interleukin-6, leukemia inhibitory factor (LIF), oncostatin M, interleukin-11, and cardiotrophin-1 (1-4). These structurally related cytokines utilize a common signal transducing subunit, gp130 (5-9). Cytokine-receptor interaction induces homo-or heterodimerization of gp130, leading to tyrosine phosphorylation of a number of intracellular substrates (10, 11). The transmembrane components of the CNTF receptor, gp130 and LIFR␤, have no intrinsic kinase activity (9, 12) but associate with the Jak/Tyk tyrosine kinases (13-15). Activation of these kinases by ligandinduced receptor dimerization is thought to initiate signal transduction and induction of gene expression (13,16,17).Cytokine stimulation induces members of the STAT transcription factor family, Stat1 and Stat3, to "dock" onto receptor phosphotyrosines, enabling their own tyrosine phosphorylation (17)(18)(19)(20). Subsequently, STAT proteins translocate to the nucleus and bind to conserved genomic regulatory sequences to provide a rapid means of activating gene transcription (21,22). These cytokines also activate components of other intracellular signaling pathways including Ras, mitogen-activated protein kinase (MAPK), and the Fos-Jun transcription factors (7,(23)(24)(25)(26)(27). How Jak-Stat activation interacts with other signaling pathways and transcription factors to regulate cytokine-mediated transcription is poorly understood.CNTF and LIF induce expression of the neuropeptide vasoactive intestinal peptide (VIP) in sympathetic neurons in culture (28, 29) and in a human neuroblastoma cell line, NBFL (30, 31). To gain insight into the mechanisms underlying cytokine-mediated activation of neuronal gene expression, we have investigated the genomic regulatory elements mediating the inductio...