Analysis of the networks controlling the antimicrobial-peptide-dependent induction of Klebsiella pneumoniae virulence factors

Antimicrobial peptides (APs) impose a threat to the survival of pathogens, and it is reasonable to postulate that bacteria have developed strategies to counteract them. Polymyxins are becoming the last resort to treat infections caused by multidrug-resistant Gram-negative bacteria and, similar to APs, they interact with the anionic lipopolysaccharide. Given that polymyxins and APs share the initial target, it is possible that bacterial defense mechanisms against polymyxins will be also effective against host APs. We sought to determine whether exposure to polymyxin will increase Klebsiella pneumoniae resistance to host APs. Indeed, exposure of K. pneumoniae to polymyxin induces cross-resistance not only to polymyxin itself but also to APs present in the airways. Polymyxin treatment upregulates the expression of the capsule polysaccharide operon and the loci required to modify the lipid A with aminoarabinose and palmitate with a concomitant increase in capsule and lipid A species containing such modifications. Moreover, these surface changes contribute to APs resistance and also to polymyxin-induced cross-resistance to APs. Bacterial loads of lipid A mutants in trachea and lungs of intranasally infected mice were lower than those of wild-type strain. PhoPQ, PmrAB, and the Rcs system govern polymyxin-induced transcriptional changes, and there is a cross talk between PhoPQ and the Rcs system. Our findings support the notion that Klebsiella activates a defense program against APs that is controlled by three signaling systems. Therapeutic strategies directed to prevent the activation of this program could be a new approach worth exploring to facilitate the clearance of the pathogen from the airways.

The spark of life:The role of electric fields in regulating cell behaviour using the eye as a model system

Endogenous electric fields (EF) have long been known to influence cell behaviour during development, neural cell tropism, wound healing and cell behaviour generally. The effect is based on short circuiting of electrical potential differences across cell and tissue boundaries generated by ionic segregation. Recent in vitro and in vivo studies have shown that EF regulate not only cell movement but orientation of cells during mitosis, an effect which may underlie shaping of tissues and organs. The molecular basis of this effect is founded on receptor-mediated cell signalling events and alterations in cytoskeletal function as revealed in studies of gene deficient cells. Remarkably, not all cells respond directionally to EF in the same way and this has consequences, for instance, for lens development and vascular remodelling. The physical basis of EF effect may be related to changes induced in 'bound water' at the cell surface, whose organisation in association with trans-membrane proteins (e.g. receptors) is disrupted when EF are generated. Copyright © 2007 S. Karger AG.

An Integrated Sensing System for Monitoring the Corrosion Activity in Concrete Blocks Exposed at Hangzhou Bay Bridge

This paper discusses the importance of integrated sensing systems comprising techniques that give different types of data from a structure exposed to the marine environment so that its service life could reliably be predicted. For this purpose, a novel sensor combination was designed and installed in concrete panels which were exposed to Hangzhou Bay Bridge in China. The integrated sensor probe was used to monitor the cover concrete as well as the reinforcement. The sensor probes were connected to a monitoring station, which enabled access and control of the data remotely from Belfast, UK. The initial data obtained from the monitoring station gives interesting information on the early age properties of concrete and distinct variations in these properties with different types of concrete. This paper also reports the variation in electrical properties of different concrete samples and environmental data in response to the marine exposure condition at Hangzhou bay bridge.

Sexual offences against children: An exploration of attrition in the Northern Ireland criminal justice system

Objective: This paper uses data provided by the Police Service for Northern Ireland (PSNI) to compare the characteristics and outcomes of reported sexual offences involving child and adult victims and explore the factors associated with case outcomes.
Method: PSNI provided data on 8,789 sexual offences recorded between April 2001 and March 2006. Case outcomes were based on whether a case was recorded by police as having sufficient evidence to summons, charge, or caution an offender (detected). Where an offender was summonsed, charged, or cautioned, this is classified as detection with a formal sanction. A case can also be classified as "detected" without a formal sanction. The analysis focused on two key categories of detection without formal sanction: cases in which the police deem there to be sufficient evidence to charge an offender but took no further action because the victim did not wish to prosecute, or because the police or the Public Prosecution Service (PPS) decided that no useful purpose would be served by proceeding.
Results: The analysis confirmed that the characteristics of recorded sexual offences involving adult and child victims vary significantly according to gender, offence type, the timing of report and victim-offender relationship. Almost half of child sex abuse cases are not detected by police and a quarter do not proceed through the criminal justice system because either the victim declines to prosecute or the Police/PPS decide not to proceed. Only one in five child cases involved detection with a formal sanction. Child groups with lower detection with formal sanction rates included children under 5, teenagers, those who do not report when the abuse occurs but disclose at a later date; and those who experience abuse at the hands of peers and adults known to them but not related. The analysis also highlighted variation in formal sanction rates depending on where the offence was reported.
Conclusions: Consideration needs to be given to improving the criminal justice response to specific child groups as well as monitoring detection rates in different police areas in order to address potential practice variation.
Practice implications: Consideration needs to be given to improving the professional response in relation to with particularly lower detection with formal sanction rates. There is also a need to monitor case outcomes to ensure that child victims in different areas receive a similar service.

Lymphocyte subsets in conjunctival mucosa-associated-lymphoid-tissue after exposure to retinal-S-antigen

Purpose: To evaluate the immune cell subsets in conjunctival mucosa-associated-lymphoid-tissue (C-MALT) following challenge with antigen. Methods: Ten adult female Lewis rats were studied. Five rats received one drop (5 µL) of retinal S-antigen (500 µg/mL in phosphate buffered saline, PBS) instilled into the lower fornix twice daily for 10 consecutive days. Five rats received PBS only and served as controls for the experiment. Two days after the last instillation the animals were sacrificed and the orbital contents prepared for immunohistological staining. A panel of monoclonal antibodies was used: CD5, CD4, CD8, CD25, and CD45RA. The number of positive cells were counted in sections of epibulbar, forniceal, and tarsal conjunctiva. Results: There was a significant increase in the number of CD8 T lymphocytes in the conjunctiva of animals receiving retinal S-antigen when compared to control animals. Conclusion: Conjunctival instillation of retinal S-antigen causes an immune response in the C-MALT with a significant increase in the CD8 T lymphocyte subset in this tissue. This response may be involved in the induction of tolerance to the encountered antigen.

Multi-detection of Paralytic, Diarrheic and Amnesic Shellfish Toxins by an Inhibition Immunoassay Using a Microsphere-Flow Cytometry System

The presence of paralytic shellfish poisoning (PSP), diarrheic shellfish poisoning (DSP) and amnesic shellfish poisoning (ASP) toxins in seafood is a severe and growing threat to human health. In order to minimize the risks of human exposure, the maximum content of these toxins in seafood has been limited by legal regulations worldwide. The regulated limits are established in equivalents of the main representatives of the groups: saxitoxin (STX), okadaic acid (OA) and domoic acid (DA), for PSP, DSP and ASP, respectively. In this study a multi-detection method to screen shellfish samples for the presence of these toxins simultaneously was developed. Multiplexing was achieved using a solid-phase microsphere assay coupled to flow-fluorimetry detection, based on the Luminex xMap technology. The multi-detection method consists of three simultaneous competition immunoassays. Free toxins in solution compete with STX, OA or DA immobilized on the surface of three different classes of microspheres for binding to specific monoclonal antibodies. The IC50 obtained in buffer was similar in single- and multi-detection: 5.6 ± 1.1 ng/mL for STX, 1.1 ± 0.03 ng/mL for OA and 1.9 ± 0.1 ng/mL for DA. The sample preparation protocol was optimized for the simultaneous extraction of STX, OA and DA with a mixture of methanol and acetate buffer. The three immunoassays performed well with mussel and scallop matrixes displaying adequate dynamic ranges and recovery rates (around 90 % for STX, 80 % for OA and 100 % for DA). This microsphere-based multi-detection immunoassay provides an easy and rapid screening method capable of detecting simultaneously in the same sample three regulated groups of marine toxins.

First results on cell irradiation with laser-driven protons on the TARANIS system

The ultra short duration of laser-driven multi-MeV ion bursts offers the possibility of radiobiological studies at extremely high dose rates. Employing the TARANIS Terawatt laser at Queen's University, the effect of proton irradiation at MeV-range energies on live cells has been investigated at dose rates exceeding 10 Gy/s as a single exposure. A clonogenic assay showed consistent lethal effects on V-79 live, cells, which, even at these dose rates, appear to be in line with previously published results employing conventional sources. A Relative Biological Effectiveness (RBE) of 1.4±0.2 at 10% survival is estimated from a comparison with a 225 kVp X-ray source.

Interaction of combined chloride and sulfate ingression in concrete

Chloride is the most severe form of deterioration experienced by concrete and one of the principal sources of chlorides is sea water. However, the presence of sulfates in seawater will influence the movement of chloride ions and vice versa. This interaction is not well understood and current codes of practice provide no guidelines for such dual exposure.
An investigation to monitor combined effect of the ingress of chlorides and sulfates during a realistic 12 month wetting and drying exposure regime to simulate conditions in which multiple mode transport mechanisms are active was conducted on a variety of binders (PC, PFA and GGBS). Penetration was evaluated using water and acid soluble chloride profiles and sulfate profiles.
It was found that the nature of the exposure provided multiple modes of transport within the concrete, thus creating a complex pattern of distribution of ions. The presence of sulfates decreased the penetration of chlorides in the PC system at all ages relative to a chloride only control. The matrices containing PFA and GGBS also showed an initial decrease in chloride penetration. However, after six months the presence of sulfates then increased chloride penetration.

Putrescine Reduces Antibiotic-Induced Oxidative Stress as a Mechanism of Modulation of Antibiotic Resistance in Burkholderia cenocepacia

Communication of antibiotic resistance among bacteria via small molecules is implicated in transient reduction of bacterial susceptibility to antibiotics, which could lead to therapeutic failures aggravating the problem of antibiotic resistance. Released putrescine from the extremely antibiotic resistant bacterium Burkholderia cenocepacia protects less resistant cells from different species against the antimicrobial peptide polymyxin B (PmB). Exposure of B. cenocepacia to sub-lethal concentrations of PmB and other bactericidal antibiotics induce reactive oxygen species (ROS) production and expression of the oxidative stress response regulator OxyR. We evaluated whether putrescine alleviates antibiotic-induced oxidative stress. The accumulation of intracellular ROS such as superoxide ion and hydrogen peroxide was assessed fluorometrically with dichlorofluorescein diacetate, while the expression of OxyR and putrescine synthesis enzymes was determined in luciferase assays using chromosomal promoter-lux reporter system fusions. We evaluated wild type and isogenic deletion mutant strains with defects in putrescine biosynthesis after exposure to sub-lethal concentrations of PmB and other bactericidal antibiotics. Exogenous putrescine protected against oxidative stress induced by PmB and other antibiotics, whereas reduced putrescine synthesis resulted in increased ROS generation, and a parallel increased sensitivity to PmB. Of the 3 B. cenocepacia putrescine synthesizing enzymes, PmB induced only BCAL2641, an ornithine decarboxylase. This study exposes BCAL2641 as a critical component of the putrescine-mediated communication of antibiotic resistance, and as a plausible target for designing inhibitors that would block the communication of such resistance among different bacteria, ultimately reducing the window of therapeutic failure in treating bacterial infections.

Does wave exposure determine the interactive effects of losing key grazers and ecosystem engineers?

The consequences of biodiversity loss in the face of environmental change remain difficult to predict, given the complexity of interactions among species and the context-dependency of their functional roles within ecosystems. Predictions may be enhanced by studies testing how the interactive effects of species loss from different functional groups vary with important environmental drivers. On rocky shores, limpets and barnacles are recognised as key grazers and ecosystem engineers, respectively. Despite the large body of research examining the combined effects of limpet and barnacle removal, it is unclear how their relative importance varies according to wave exposure, which is a dominant force structuring intertidal communities. We tested the responses of algal communities to the removal of limpets and barnacles on three sheltered and three wave-exposed rocky shores on the north coast of Ireland. Limpet removal resulted in a relative increase in microalgal biomass on a single sheltered shore only, but led to the enhanced accumulation of ephemeral macroalgae on two sheltered shores and one exposed shore. On average, independently of wave exposure or shore, ephemeral macroalgae increased in response to limpet removal, but only when barnacles were removed. On two sheltered shores and one exposed shore, however, barnacles facilitated the establishment of fucoid macroalgae following limpet removal. Therefore, at the scale of this study, variability among individual shores was more important than wave exposure per se in determining the effect of limpet removal and its interaction with that of barnacles. Overall, these findings demonstrate that the interactive effects of losing key species from different functional groups may not vary predictably according to dominant environmental factors.

System level DSP synthesis using voltage overscaling, unequal error protection & adaptive quality tuning

In this paper, we propose a system level design approach considering voltage over-scaling (VOS) that achieves error resiliency using unequal error protection of different computation elements, while incurring minor quality degradation. Depending on user specifications and severity of process variations/channel noise, the degree of VOS in each block of the system is adaptively tuned to ensure minimum system power while providing "just-the-right" amount of quality and robustness. This is achieved, by taking into consideration system level interactions and ensuring that under any change of operating conditions only the "lesscrucial" computations, that contribute less to block/system output quality, are affected. The design methodology applied to a DCT/IDCT system shows large power benefits (up to 69%) at reasonable image quality while tolerating errors induced by varying operating conditions (VOS, process variations, channel noise). Interestingly, the proposed IDCT scheme conceals channel noise at scaled voltages. ©2009 IEEE.

The Kepler-10 Planetary System Revisited by HARPS-N: A Hot Rocky World and a Solid Neptune-Mass Planet

Kepler-10b was the first rocky planet detected by the Kepler satellite and confirmed with radial velocity follow-up observations from Keck-HIRES. The mass of the planet was measured with a precision of around 30%, which was
insufficient to constrain models of its internal structure and composition in detail. In addition to Kepler-10b, a second planet transiting the same star with a period of 45 days was statistically validated, but the radial velocities were only
good enough to set an upper limit of 20 M⊕ for the mass of Kepler-10c. To improve the precision on the mass for planet b, the HARPS-N Collaboration decided to observe Kepler-10 intensively with the HARPS-N spectrograph
on the Telescopio Nazionale Galileo on La Palma. In total, 148 high-quality radial-velocity measurements were obtained over two observing seasons. These new data allow us to improve the precision of the mass determination for Kepler-10b to 15%. With a mass of 3.33 ± 0.49 M⊕ and an updated radius of 1.47+0.03 −0.02 R⊕, Kepler-10b has a density of 5.8 ± 0.8 g cm−3, very close to the value predicted by models with the same internal structure and composition as the Earth. We were also able to determine a mass for the 45-day period planet Kepler-10c, with an even better precision of 11%. With a mass of 17.2 ± 1.9 M⊕ and radius of 2.35+0.09 −0.04 R⊕, Kepler-10c has a density of 7.1 ± 1.0 g cm−3. Kepler-10c appears to be the first strong evidence of a class of more massive solid planets with longer orbital periods

The performance of a cathodic protection system in reinforced concrete structure: Monitoring and Service Life Modelling

The use of cathodic protection in reinforced concrete is becoming increasingly common with such systems being installed on a number of structures throughout the United Kingdom and Ireland. However the prescribed design lives (or service life) of each cathodic protection system vary widely. The aim of this project was to assess the effectiveness of a sacrificial anode cathodic protection system and to predict its design life through a series of laboratory based experiments. The experimental plan involved casting a number of slabs which represented a common road bridge structure. The corrosion of the steel within the experimental slabs was then accelerated prior to installation of a cathodic protection system. During the experiment corrosion potential of the steel reinforcement was monitored using half-cell measurement. Additionally the current flow between the cathodic protection system and the steel reinforcement was recorded to assess the degree of protection. A combination of theoretical calculations and experimental results were then collated to determine the design life of this cathodic protection system. It can be concluded that this sacrificial anode based cathodic protection system was effective in halting the corrosion of steel reinforcement in the concrete slabs studied. Both the corrosion current and half-cell potentials indicated a change in passivity for the steel reinforcement once sacrificial anodes were introduced. The corrosion current was observed to be sensitive to the changes to the exposure environment. Based on the experimental variables studied the design life of this sacrificial anode can be taken as 26 to 30 years.

More nonlocality with less entanglement in a tripartite atom-optomechanical system

Quantum effects in hybrid atomic optomechanics in a system comprising a cloud of atoms and a mobile mirror mediated by a single-mode cavity are studied. Tripartite non-locality is observed in the atom-light-mirror system, as demonstrated by the violation of the Mermin-Klyshko (MK) inequality. It has been shown [C. Genes, et al., PRA 77, 050307 (R) (2008)] that tripartite entanglement is optimized when the cavity is resonant with the anti-Stokes sideband of the driving laser and the atomic frequency matches the Stokes one. However, we show that this is not the case for the nonlocality. The MK function achieves minima when the atoms are resonant with both the Stokes and anti-Stokes sidebands, and unexpectedly, we find violation of the MK inequality only in a parameter region where entanglement is far from being maximum. A negative relation exists between nonlocality and entanglement with consideration of the possibility of bipartite nonlocality in the violation of the MK inequality. We also study the non-classicality of the mirror by post-selected measurements, e.g. Geiger-like detection, on the cavity and/or the atoms. We show that with feasible parameters Geiger-like detection on the atoms can effectively induce mechanical non-classicality.

A comparison of the effectiveness of TiO2 photocatalysis and UVA photolysis for the destruction of three pathogenic micro-organisms

TiO₂ photocatalysis has demonstrated efficacy as a treatment process for water contaminated with chemical pollutants. When exposed to UVA light TiO₂ also demonstrates an effective bactericidal activity. The mechanism of this process has been reported to involve attack by valence band generated hydroxyl radicals. In this study when three common bacterial pathogens, Escherichia coli, Salmonella enterica serovar Enteritidis and Pseudomonas aeruginosa, were exposed to TiO₂ and UVA light a substantial decrease in bacterial numbers was observed. Control experiments in which all three pathogens were exposed to UVA light only resulted in a similar reduction in bacterial numbers. Moreover, exposure to UVA light alone resulted in the production of a smaller than average colony phenotype among the surviving bacteria, for all three pathogens examined, a finding which was not observed following treatment with UVA and TiO₂. Small slow growing colonies have been described for several pathogenic bacteria and are referred to as small colony variants. Several studies have demonstrated an association between small colony variants and persistent, recurrent and antibiotic resistant infections. We propose that the production of small colony variants of pathogenic bacteria following UVA treatment of drinking water may represent a health hazard. As these small colony variants were not observed with the UVA/TiO₂ system this potential hazard is not a risk when using this technology. It would also appear that the bactericidal mechanism is different with the UVA/TiO₂ process compared to when UVA light is used alone.