Comparison of plasma transport theory with measurements in a H- discharge

This paper and its companion paper describe the comparison between a one-dimensional theoretical model of a hydrogen discharge in a magnetic multipole plasma source and experimental measurements of the plasma parameters. The discharge chamber, described here, has been designed to produce significant densities of H- ions by incorporating a weak transverse field through the discharge to obtain electron cooling so as to maximize H- production. Langmuir probes are used to monitor the plasma, determining the ion density, the electron density and temperature and the plasma potential. The negative density is measured by photo-detachment of the extra electron using an intense laser beam. The model, described in the companion paper, uses the presented source geometry to calculate these plasma quantities as a function of the major are parameters; namely the are current and voltage and gas pressure. Good agreement is obtained between theory and experiment as a function of position and arc parameters.

ELECTRON-ENERGY DISTRIBUTION-FUNCTIONS AND NEGATIVE-ION CONCENTRATIONS IN TANDEM AND HYBRID MULTICUSP NEGATIVE HYDROGEN-ION SOURCES

The second derivative of a Langmuir probe characteristic is used to establish the electron energy distribution function (EEDF) in both a tandem and hybrid multicusp H- ion source. Moveable probes are used to establish the spatial variation of the EEDF. The negative ion density is measured by laser induced photo-detachment. In the case of the hybrid source the EEDF consists of a cold Maxwellian in the central region of the source; the electron temperature increases with increasing discharge current (rising from 0.3 eV at 1 A to 1.2 eV at 50 A when the pressure is 0.4 Pa). A hot-electron tail exists in the EEDF of the driver region adjacent to each filament which is shown to consist of a distinct group of primary electrons at low pressure (0.08 Pa) but becomes degraded mainly through inelastic collisions at higher pressures (0.27 Pa). The tandem source, on the other hand, has a single driver region which extends throughout the central region. The primary electron confinement times are much longer so that even at the lowest pressure considered (0.07 Pa) the primaries are degraded. In both cases the measured EEDF at specific locations and values of discharge operating parameters are used to establish the rate coefficients for the processes of importance in H- production and destruction.

Detection of Voigt Spectral Line Profiles of Hydrogen Radio Recombination Lines toward Sagittarius B2(N)

We report the detection of Voigt spectral line profiles of radio recombination lines (RRLs) toward Sagittarius B2(N) with the 100 m Green Bank Telescope (GBT). At radio wavelengths, astronomical spectra are highly populated with RRLs, which serve as ideal probes of the physical conditions in molecular cloud complexes. An analysis of the Hn alpha lines presented herein shows that RRLs of higher principal quantum number (n > 90) are generally divergent from their expected Gaussian profiles and, moreover, are well described by their respective Voigt profiles. This is in agreement with the theory that spectral lines experience pressure broadening as a result of electron collisions at lower radio frequencies. Given the inherent technical difficulties regarding the detection and profiling of true RRL wing spans and shapes, it is crucial that the observing instrumentation produce flat baselines as well as high-sensitivity, high-resolution data. The GBT has demonstrated its capabilities regarding all of these aspects, and we believe that future observations of RRL emission via the GBT will be crucial toward advancing our knowledge of the larger-scale extended structures of ionized gas in the interstellar medium (ISM).

Development and comparison of a Primer-Probe Energy Transfer based assay and a 5 ' conjugated Minor Groove Binder assay for sensitive real-time PCR detection of infectious laryngotracheitis virus

In this study the design and development of two real-time PCR assays for the rapid, sensitive and specific detection of infectious laryngotracheitis virus (ILTV) DNA is described. A Primer-Probe Energy Transfer (PriProET) assay and 5' conjugated Minor Groove Binder (MGB) method are compared and contrasted. Both have been designed to target the thymidine kinase gene of the ILTV genome. Both PriProET and MGB assays are capable of detecting 20 copies of a DNA standard per reaction and are linear from 2 x 10(8) to 2 x 10(2) copies/mu l. Neither PriProET, nor MGB reacted with heterologous herpesviruses, indicating a high specificity of the two methods as novel tools for virus detection and identification. This study demonstrates the suitability of PriProET and 5' conjugated MGB probes as real-time PCR chemistries for the diagnosis of respiratory diseases caused by ILTV. (C) 2011 Elsevier B.V. All rights reserved.

Development of a minor groove binder assay for real-time one-step RT-PCR detection of swine vesicular disease virus

The design and development of a 5' conjugated minor groove binder (MGB) probe real-time RT-PCR assay are described for rapid, sensitive and specific detection of swine vesicular disease virus (SVDV) RNA. The assay is designed to target the 2C gene of the SVDV genome and is capable of detecting 2 x 10(2) copies of an RNA standard per reaction. It does not detect any of the other RNA viruses that cause vesicular disease in pigs, or the human enterovirus, Coxsackie B5 virus (CVB5) which is closely related antigenically to SVDV. The linear range of this test was from 2 x 10(2) to 2 x 10(8) copies/mu l. The assay is rapid and can detect SVDV RNA in just over 3.5 h including the time required for nucleic acid extraction. The development of this assay provides a useful tool for the differential diagnosis of SVD or for the detection of SVDV in research applications. This study demonstrates the suitability of MGB probes as a real-time PCR chemistry for the diagnosis of swine vesicular disease. (C) 2010 Elsevier B.V. All rights reserved.

Validation of membrane protein topology models by oxidative labeling and mass spectrometry

Computer-assisted topology predictions are widely used to build low-resolution structural models of integral membrane proteins (IMPs). Experimental validation of these models by traditional methods is labor intensive and requires modifications that might alter the IMP native conformation. This work employs oxidative labeling coupled with mass spectrometry (MS) as a validation tool for computer-generated topology models. ·OH exposure introduces oxidative modifications in solvent-accessible regions, whereas buried segments (e.g., transmembrane helices) are non-oxidizable. The Escherichia coli protein WaaL (O-antigen ligase) is predicted to have 12 transmembrane helices and a large extramembrane domain (Pérez et al., Mol. Microbiol. 2008, 70, 1424). Tryptic digestion and LC-MS/MS were used to map the oxidative labeling behavior of WaaL. Met and Cys exhibit high intrinsic reactivities with ·OH, making them sensitive probes for solvent accessibility assays. Overall, the oxidation pattern of these residues is consistent with the originally proposed WaaL topology. One residue (M151), however, undergoes partial oxidation despite being predicted to reside within a transmembrane helix. Using an improved computer algorithm, a slightly modified topology model was generated that places M151 closer to the membrane interface. On the basis of the labeling data, it is concluded that the refined model more accurately reflects the actual topology of WaaL. We propose that the combination of oxidative labeling and MS represents a useful strategy for assessing the accuracy of IMP topology predictions, supplementing data obtained in traditional biochemical assays. In the future, it might be possible to incorporate oxidative labeling data directly as constraints in topology prediction algorithms.

Intracellular survival of Burkholderia cenocepacia in macrophages is associated with a delay in the maturation of bacteria-containing vacuoles

Strains of the Burkholderia cepacia complex (Bcc) are opportunistic bacteria that can cause life-threatening infections in patients with cystic fibrosis and chronic granulomatous disease. Previous work has shown that Bcc isolates can persist in membrane-bound vacuoles within amoeba and macrophages without bacterial replication, but the detailed mechanism of bacterial persistence is unknown. In this study, we have investigated the survival of the Burkholderia cenocepacia strain J2315 within RAW264.7 murine macrophages. Strain J2315 is a prototypic isolate of the widespread and transmissible ET12 clone. Unlike heat-inactivated bacteria, which reach lysosomes shortly after internalization, vacuoles containing live B. cenocepacia J2315 accumulate the late endosome/lysosome marker LAMP-1 and start fusing with lysosomal compartments only after 6 h post internalization. Using fluorescent fluid-phase probes, we also demonstrated that B. cenocepacia-containing vacuoles continued to interact with newly formed endosomes, and maintained a luminal pH of 6.4 +/- 0.12. In contrast, vacuoles containing heat-inactivated bacteria had an average pH of 4.8 +/- 0.03 and rapidly merged with lysosomes. Additional experiments using concanamycin A, a specific inhibitor of the vacuolar H+-ATPase, revealed that vacuoles containing live bacteria did not exclude the H+-ATPase. This mode of bacterial survival did not require type III secretion, as no differences were found between wild type and a type III secretion mutant strain. Collectively, our results suggest that intracellular B. cenocepacia cause a delay in the maturation of the phagosome, which may contribute to facilitate bacterial escape from the microbicidal activities of the host cell.

Relatedness of O-specific lipopolysaccharide side chain genes from strains of Shigella boydii type 12 belonging to two clonal groups and from Escherichia coli O7:K1

The O-specific lipopolysaccharide side chains of Escherichia coli O7 and Shigella boydii type 12 possess similar but not identical chemical structures. We investigated the genetic relatedness between the O-specific side chain genes in members of these two species. Examination of outer membrane protein and lipopolysaccharide (LPS) banding patterns demonstrated that five strains which had been identified as S. boydii type 12 fell into two clonal groups, SB1 and SB2. Hybridizations with O7-specific radiolabeled probes derived from the chromosomal DNA of an E. coli O7 strain detected identical fragments among the three SB1 strains of S. boydii type 12 and the two E. coli O7 reference isolates. The two other S. boydii type 12 strains, which belonged to the SB2 clone, did not show homologies with the O7 probe under high-stringency conditions of hybridization. The homology between the O7 and type 12 LPS gene regions from the SB1 strains was further confirmed by the construction of O-specific side chain-deficient mutations in these strains by homologous recombination of a suicide plasmid containing O7-specific DNA sequences. Immunoblot experiments with O7 antiserum gave a weak cross-reaction with LPS purified from the SB2 strains but a very strong cross-reaction with the LPS from SB1 isolates. Antiserum raised to one of the SB2 strains cross-reacted only with S. boydii type 12 LPS from the SB1 clone but failed to react with O7 LPS.

Chymotrypsin-like serine proteinases are involved in the maintenance of cell viability

An increasing number of studies have implicated serine proteinases in the development of apoptosis. In this study, we assessed the ability of a set of highly specific irreversible inhibitors (activity probes), incorporating an a-amino alkane diphenyl phosphonate moiety, to modulate cell death. In an initial assessment of the cellular toxicity of these activity probes, we discovered that one example, N-a-tetramethylrhodamine phenylalanine diphenylphosphonate {TMR-Phe^P(OPh)₂} caused a concentration-dependent decrease in the viability of HeLa and U251 mg cells. This reduced cell viability was associated with a time-dependent increase in caspase-3 activity, PARP cleavage and phosphatidylserine translocation, establishing apoptosis as the mechanism of cell death. SDS-PAGE analysis of cell lysates prepared from the HeLa cells treated with TMR-Phe^P(OPh)₂, revealed the presence of a fluorescent band of molecular weight 58 kDa. Given that we have previously reported on the use of this type of activity probe to reveal active proteolytic species, we believe that we have identified a chymotrypsin-like serine proteinase activity integral to the maintenance of cell viability.

The electrical characterization and response to hydrogen of Schottky diodes with a resistive metal electrode—rectifying an oversight in Schottky diode investigation

Schottky-barrier structures with a resistive metal electrode are examined using the 4-point probe method where the probes are connected to the metal electrode only. The observation of a significant decrease in resistance with increasing temperature (over a range of similar to 100 K) in the diode resistance-temperature (R(D)-T) characteristic is considered due to charge carrier confinement to the metal electrode at low temperature (high resistance), with the semiconductor progressively opening up as a parallel current carrying channel (low resistance) with increasing temperature due to increasing thermionic emission across the barrier. A simple model is constructed, based on thermionic emission at quasi-zero bias, that generates good fits to the experimental data. The negative differential resistance (NDR) region in the R(D)-T characteristic is a general effect and is demonstrated across a broad temperature range for a variety of Schottky structures grown on Si-, GaAs- and InP-substrates. In addition the NDR effect is harnessed in micro-scaled Pd/n-InP devices for the detection of low levels of hydrogen in an ambient atmosphere of nitrogen.

Predicting organic acid concentration from UV/vis spectrometry measurements - A comparison of machine learning techniques

The concentration of organic acids in anaerobic digesters is one of the most critical parameters for monitoring and advanced control of anaerobic digestion processes. Thus, a reliable online-measurement system is absolutely necessary. A novel approach to obtaining these measurements indirectly and online using UV/vis spectroscopic probes, in conjunction with powerful pattern recognition methods, is presented in this paper. An UV/vis spectroscopic probe from S::CAN is used in combination with a custom-built dilution system to monitor the absorption of fully fermented sludge at a spectrum from 200 to 750 nm. Advanced pattern recognition methods are then used to map the non-linear relationship between measured absorption spectra to laboratory measurements of organic acid concentrations. Linear discriminant analysis, generalized discriminant analysis (GerDA), support vector machines (SVM), relevance vector machines, random forest and neural networks are investigated for this purpose and their performance compared. To validate the approach, online measurements have been taken at a full-scale 1.3-MW industrial biogas plant. Results show that whereas some of the methods considered do not yield satisfactory results, accurate prediction of organic acid concentration ranges can be obtained with both GerDA and SVM-based classifiers, with classification rates in excess of 87% achieved on test data.

Electrical Based Sensors and Remote Monitoring System for Assessing the Corrosion Related Activity in Concrete at Hangzhou Bay Bridge

Concrete structures in marine environments are subjected to cyclic wetting and drying, corrosion of reinforcement due to chloride ingress and biological deterioration. In order to assess the quality of concrete and predict the corrosion activity of reinforcing steel in concrete in this environment, it is essential to monitor the concrete continuously right from the construction phase to the end of service life of the structure. In this paper a novel combination of sensor techniques which are integrated in a sensor probe is used to monitor the quality of cover concrete and corrosion of the reinforcement. The integrated sensor probe was embedded in different concrete samples exposed to an aggressive marine environment at the Hangzhou Bay Bridge in China. The sensor probes were connected to a monitoring station, which enabled the access and control of the data remotely from Belfast, UK. The initial data obtained from the monitoring station reflected the early age properties of the concretes and distinct variations in these properties were observed with different concrete types.

Wino's vanitas:Ivresse transgressive et affirmation littéraire dans les romans noirs de Ken Bruen

Based on a theoretical framework owing to Bourdieu, Viala and Meizoz successive adjustments of the notion of literary posturing, and on Maingueneau's concept of auctorial scenography, this chapter probes the writer's ethos of Irish crime fiction author Ken Bruen and its impact on his reception in the French literary field. It studies in particular the double transgression characteristic of his ethos as a pioneer of the Irish noir : a cultural transgression attacking relentlessly the international currency of myths and stereotypes of Ireland, and a generic transgression, which thrives in the tension between codes and constraints of the noir novel.

Rapid differentiation between fluconazole-sensitive and -resistant species of Candida directly from positive blood-culture bottles by real-time PCR

In view of both the delay in obtaining identification by conventional methods following blood-culture positivity in patients with candidaemia and the close relationship between species and fluconazole (FLC) susceptibility, early speciation of positive blood cultures has the potential to influence therapeutic decisions. The aim was to develop a rapid test to differentiate FLC-resistant from FLC-sensitive Candida species. Three TaqMan-based real-time PCR assays were developed to identify up to six Candida species directly from BacT/Alert blood-culture bottles that showed yeast cells on Gram staining at the time of initial positivity. Target sequences in the rRNA gene complex were amplified, using a consensus two-step PCR protocol, to identify Candida albicans, Candida parapsilosis, Candida tropicalis, Candida dubliniensis, Candida glabrata and Candida krusei; these are the most commonly encountered Candida species in blood cultures. The first four of these (the characteristically FLC-sensitive group) were identified in a single reaction tube using one fluorescent TaqMan probe targeting 1 8S rRNA sequences conserved in the four species. The FLC-resistant species C. krusei and C. glabrata were detected in two further reactions, each with species-specific probes. This method was validated with clinical specimens (blood cultures) positive for yeast (n=33 sets) and the results were 100% concordant with those of phenotypic identification carried out concomitantly. The reported assay significantly reduces the time required to identify the presence of C. glabrata and C. krusei in comparison with a conventional phenotypic method, from ~72 to

Acidity Characterization of Amorphous Silica-Alumina

Surface characterization of amorphous silica-alumina (ASA) by COads IR, pyridine(ads) IR, alkylamine temperature-programmed desorption (TPD), Cs+ and Cu(EDA)(2)(2+) exchange, H-1 NMR, and m-xylene isomerization points to the presence of a broad range of Bronsted and Lewis acid sites. Careful interpretation of IR spectra of adsorbed CO or pyridine confirms the presence of a few very strong Bronsted acid sites (BAS), typically at concentrations lower than 10 mu mol/g. The general procedure for alkylamine TPD, which probes both Bronsted and Lewis acidity, is modified to increase the selectivity to strong Bronsted acid sites. Poisoning of the m-xylene isomerization reaction by a base is presented as a novel method to quantify strong BAS. The surface also contains a weaker form of BAS, in concentrations between 50 and 150 mu mol/g, which can be quantified by COads IR Cu(EDA)(2)(2+) exchange also probes these sites. The structure of these sites remains unclear, but they might arise from the interaction of silanol groups with strong Lewis acid Al3+ sites. The surface also contains nonacidic aluminol and silanol sites (200-400 mu mol/g) and two forms of Lewis acid sites: (i) a weaker form associated with segregated alumina domains containing five-coordinated Al, which make up the interface between these domains and the ASA phase and (ii) a stronger form, which are undercoordinated Al sites grafted onto the silica surface. The acid catalytic activity in bifunctional n-heptane hydroconversion correlates with the concentration of strong BAS. The influence of the support electronegativity on the neopentane hydrogenolysis activity of supported Pt catalysts is considerably larger than that of the support Bronsted acidity. It is argued that strong Lewis acid sites, which are present in ASA but not in gamma-alumina, are essential to transmit the Sanderson electronegativity of the oxide support to the active Pt phase.