The Cell-Cycle Regulatory Protein p21CIP1/WAF1 Is Required for Cytolethal Distending Toxin (Cdt)-Induced Apoptosis.

The Aggregatibacter actinomycetemcomitans cytolethal distending toxin (Cdt) induces lymphocytes to undergo cell-cycle arrest and apoptosis; toxicity is dependent upon the active Cdt subunit, CdtB. We now demonstrate that p21CIP1/WAF1 is critical to Cdt-induced apoptosis. Cdt induces increases in the levels of p21CIP1/WAF1 in lymphoid cell lines, Jurkat and MyLa, and in primary human lymphocytes. These increases were dependent upon CdtB's ability to function as a phosphatidylinositol (PI) 3,4,5-triphosphate (PIP3) phosphatase. It is noteworthy that Cdt-induced increases in the levels of p21CIP1/WAF1 were accompanied by a significant decline in the levels of phosphorylated p21CIP1/WAF1. The significance of Cdt-induced p21CIP1/WAF1 increase was assessed by preventing these changes with a two-pronged approach; pre-incubation with the novel p21CIP1/WAF1 inhibitor, UC2288, and development of a p21CIP1/WAF1-deficient cell line (Jurkatp21-) using clustered regularly interspaced short palindromic repeats (CRISPR)/cas9 gene editing. UC2288 blocked toxin-induced increases in p21CIP1/WAF1, and JurkatWT cells treated with this inhibitor exhibited reduced susceptibility to Cdt-induced apoptosis. Likewise, Jurkatp21- cells failed to undergo toxin-induced apoptosis. The linkage between Cdt, p21CIP1/WAF1, and apoptosis was further established by demonstrating that Cdt-induced increases in levels of the pro-apoptotic proteins Bid, Bax, and Bak were dependent upon p21CIP1/WAF1 as these changes were not observed in Jurkatp21- cells. Finally, we determined that the p21CIP1/WAF1 increases were dependent upon toxin-induced increases in the level and activity of the chaperone heat shock protein (HSP) 90. We propose that p21CIP1/WAF1 plays a key pro-apoptotic role in mediating Cdt-induced toxicity

Sonification Mapping Configurations: Pairings Of Real-Time Exhibits And Sound

Presented at the 19th International Conference on Auditory Display (ICAD2013) on July 6-9, 2013 in Lodz, Poland.Visitors to aquariums typically rely on their vision to interact with live exhibits that convey rich descriptive and aesthetic visual information. However, some visitors may prefer or need to have an alternative interpretation of the exhibitÕs visual scene to improve their experience. Musical sonification has been explored as an interpretive strategy for this purpose and related work provides some guidance for sonification design, yet more empirical work on developing and validating the music-to-visual scene mappings needs to be completed. This paper discusses work to validate mappings that were developed through an investigation of musician performances for two specific live animal exhibits at the Georgia Aquarium. In this proposed study, participants will provide feedback on musical mapping examples which will help inform design of a real-time sonification system for aquarium exhibits. Here, we describe our motivation, methods, and expected contributions

Hybrid mode-scattering/sound-absorbing segmented liner system and method

A hybrid mode-scattering/sound-absorbing segmented liner system and method in which an initial sound field within a duct is steered or scattered into higher-order modes in a first mode-scattering segment such that it is more readily and effectively absorbed in a second sound-absorbing segment. The mode-scattering segment is preferably a series of active control components positioned along the annulus of the duct, each of which includes a controller and a resonator into which a piezoelectric transducer generates the steering noise. The sound-absorbing segment is positioned acoustically downstream of the mode-scattering segment, and preferably comprises a honeycomb-backed passive acoustic liner. The invention is particularly adapted for use in turbofan engines, both in the inlet and exhaust

Active Control of Low-Speed Fan Tonal Noise Using Actuators Mounted in Stator Vanes: Part III Results

A test program to demonstrate simplification of Active Noise Control (ANC) systems relative to standard techniques was performed on the NASA Glenn Active Noise Control Fan from May through September 2001. The target mode was the m = 2 circumferential mode generated by the rotor-stator interaction at 2BPF. Seven radials (combined inlet and exhaust) were present at this condition. Several different error-sensing strategies were implemented. Integration of the error-sensors with passive treatment was investigated. These were: (i) an in-duct linear axial array, (ii) an induct steering array, (iii) a pylon-mounted array, and (iv) a near-field boom array. The effect of incorporating passive treatment was investigated as well as reducing the actuator count. These simplified systems were compared to a fully ANC specified system. Modal data acquired using the Rotating Rake are presented for a range of corrected fan rpm. Simplified control has been demonstrated to be possible but requires a well-known and dominant mode signature. The documented results here in are part III of a three-part series of reports with the same base title. Part I and II document the control system and error-sensing design and implementation

Lymphoid Susceptibility to the Aggregatibacter Actinomycetemcomitans Cytolethal Distending Toxin is Dependent Upon Baseline Levels of the Signaling Lipid, Phosphatidylinositol-3,4,5-Triphosphate

The Aggregatibacter actinomycetemcomitans cytolethal distending toxin (Cdt) induces G2 arrest and apoptosis in lymphocytes and other cell types. We have shown that the active subunit, CdtB, exhibits phosphatidylinositol-3,4,5-triphosphate (PIP3) phosphatase activity and depletes lymphoid cells of PIP3. Hence we propose that Cdt toxicity results from depletion of this signaling lipid and perturbation of phosphatidylinositol-3-kinase (PI-3K)/PIP3/Akt signaling. We have now focused on the relationship between cell susceptibility to CdtB and differences in the status of baseline PIP3 levels. Our studies demonstrate that the baseline level of PIP3, and likely the dependence of cells on steady-state activity of the PI-3K signaling pathway for growth and survival, influence cell susceptibility to the toxic effects of Cdt. Jurkat cells with known defects in both PIP3 degradative enzymes, PTEN and SHIP1, not only contain high baseline levels of PIP3, pAkt, and pGSK3β, but also exhibit high sensitivity to Cdt. In contrast, HUT78 cells, with no known defects in this pathway, contain low levels of PIP3, pAkt, and pGSK3β and likely minimal dependence on the PI-3K signaling pathway for growth and survival, and exhibit reduced susceptibility to Cdt. These differences in susceptibility to Cdt cannot be explained by differential toxin binding or internalization of the active subunit. Indeed, we now demonstrate that Jurkat and HUT78 cells bind toxin at comparable levels and internalize relatively equal amounts of CdtB. The relevance of these observations to the mode of action of Cdt and its potential role as a virulence factor isdiscussed. © 2016 John Wiley & Sons A/S

Measuring the Edwards-Anderson order parameter of the Bose glass : a quantum gas microscope approach

We thank D Cassettari, A Daley, S Denny, J Keeling, P Kirton and A Trombettoni for insightful discussions and assistance. Computations were performed on the EPSRC CDT Computer Cluster and the University of St Andrews School of Physics & Astronomy computer cluster. SJT acknowledges studentship funding from EPSRC under grant no. EP/G03673X/1. GDB acknowledges support from the Leverhulme Trust RPG-2013-074.With the advent of spatially resolved fluorescence imaging in quantum gas microscopes, it is now possible to directly image glassy phases and probe the local effects of disorder in a highly controllable setup. Here we present numerical calculations using a spatially resolved local mean-field theory, show that it captures the essential physics of the disordered system and use it to simulate the density distributions seen in single-shot fluorescence microscopy. From these simulated images we extract local properties of the phases which are measurable by a quantum gas microscope and show that unambiguous detection of the Bose glass is possible. In particular, we show that experimental determination of the Edwards-Anderson order parameter is possible in a strongly correlated quantum system using existing experiments. We also suggest modifications to the experiments which will allow further properties of the Bose glass to be measured.PostprintPeer reviewe

Dynamics of success and failure in phage and antibiotic therapy in experimental infections

BACKGROUND: In 1982 Smith and Huggins showed that bacteriophages could be at least as effective as antibiotics in preventing mortality from experimental infections with a capsulated E. coli (K1) in mice. Phages that required the K1 capsule for infection were more effective than phages that did not require this capsule, but the efficacies of phages and antibiotics in preventing mortality both declined with time between infection and treatment, becoming virtually ineffective within 16 hours. RESULTS: We develop quantitative microbiological procedures that (1) explore the in vivo processes responsible for the efficacy of phage and antibiotic treatment protocols in experimental infections (the Resistance Competition Assay, or RCA), and (2) survey the therapeutic potential of phages in vitro (the Phage Replication Assay or PRA). We illustrate the application and utility of these methods in a repetition of Smith and Huggins' experiments, using the E. coli K1 mouse thigh infection model, and applying treatments of phages or streptomycin. CONCLUSIONS: 1) The Smith and Huggins phage and antibiotic therapy results are quantitatively and qualitatively robust. (2) Our RCA values reflect the microbiological efficacies of the different phages and of streptomycin in preventing mortality, and reflect the decline in their efficacy with a delay in treatment. These results show specifically that bacteria become refractory to treatment over the term of infection. (3) The K1-specific and non-specific phages had similar replication rates on bacteria grown in broth (based on the PRA), but the K1-specific phage had markedly greater replication rates in mouse serum

Settlement of oysters, Crassostrea virginica (Gmelin, 1791), on oyster shell, expanded shale and tire chips in the James River, Virginia

The effectiveness of oyster shell, expanded shale, and tire chips as substrates for settlement of oysters, Crassostrea virgi11ica (Gmelin), was compared at four locations in the James River, Virginia, over three two-week time intervals in August and September, 1988. Only differences between substrate were significant (P \u3c 0.001). Over all locations and time intervals, a significantly higher (P \u3c 0.001) proportion of total oyster settlement occurred on oyster shell (63.8%) than on either tire chips (22.1 %) or expanded shale (14.2%)

Blockade of the PI-3K Signalling Pathway by the Aggregatibacter Actinomycetemcomitans Cytolethal Distending Toxin Induces Macrophages to Synthesize and Secrete Pro-Inflammatory Cytokines

The Aggregatibactor actinomycetemcomitans cytolethal distending toxin (Cdt) induces G2 arrest and apoptosis in lymphocytes; these toxic effects are due to the active subunit, CdtB, which functions as a phosphatidylinositol-3,4,5-triphosphate (PIP3) phosphatase. We now extend our investigation and demonstrate that Cdt is able to perturb human macrophage function. THP-1- and monocyte-derived macrophages were found not to be susceptible to Cdt-induced apoptosis. Nonetheless, the toxin was capable of binding to macrophages and perturbing PI-3K signalling resulting in decreased PIP3 levels and reduced phosphorylation of Akt and GSK3β these changes were accompanied by concomitant alterations in kinase activity. Exposure of monocytes and macrophages to Cdt resulted in pro-inflammatory cytokine production including increased expression and release of IL-1β, TNFα and IL-6. Furthermore, treatment of cells with either TLR-2, -3 or -4 agonists in the presence of Cdt resulted in an augmented pro-inflammatory response relative to agonist alone. GSK3β inhibitors blocked the Cdt-induced pro-inflammatory cytokine response suggesting a pivotal role for PI-3K blockade, concomitant decrease in GSK3β phosphorylation and increased kinase activity. Collectively, these studies provide new insight into the virulence potential of Cdt in mediating the pathogenesis of disease caused by Cdt-producing organisms. © 2014 John Wiley & Sons Ltd

Hearing the silences: adult Nigerian women’s accounts of ‘early marriages’

‘Early marriage’ is a relatively common, but under-researched global phenomenon, associated with poor health, mental health, educational and occupational outcomes, particularly for young girls. In this article, we draw on qualitative interviews with 6 Nigerian women from Sokoto State, who were married between the ages of 8 and 15. The interviews explored young women’s experiences of the transition to marriage, being married, pregnancy and their understanding of the marital and parental role. Using Interpretive Phenomenological Analysis, we explore women’s constrained articulations of their experiences of early marriage, as they are constituted within a social context where the identity of ‘woman’ is bound up in values and practices around marriage and motherhood. We explore the complexity of ‘hearing’ women’s experiences when their identities are bound up in culturally overdetermined ideas of femininity that function explicitly to silence and constrain the spaces in which women can speak