bluuebunny/crossref_metadata_2025 huggingface dataset:
Logical inconsistency in construct relationships: Conflict or complexity?
Authors: Winter David A.
Published: July 2011

The study examines repertory grid measures of conflict. Evidence is provided that neurotic disorder is associated with high levels of logical consistency in construct relationships, particularly in those concerning ‘symptom constructs’, but this is found to be in large part a function of a relationship between neurotic disorder and high intensity of construct relationships. No significant changes in conflict scores are found during psychological therapy, but high conflict scores are associated with clinical allocation to, and improvement in, group psychotherapy and lack of conflict to allocation to, and improvement in, behaviour therapy.
Strategies of clinical listening
Authors: Watts Fraser N.
Published: July 2011

Some of the problems that confront the clinician in seeking to comprehend patients' accounts of their problems are discussed. It must be anticipated that these will be systematically deficient, both because of limitations in patients' awareness and because of the effects of the communication situations.

Several strategies of listening that are available: (a) attention to patients' exact language, (b) heuristic strategies, (c) selective tuning, (d) relating information to emerging formulations and (e) ‘evenly suspended attention’ are discussed. The importance of the scientific investigation of clinical listening is emphasized, so that sound methods of training in listening can be developed.
Changes in patients during treatment at the Henderson Hospital therapeutic community during 1977–81
Authors: Norris Margaret
Published: July 2011

This study is the third in a series which has used repertory grids to evaluate changes in people in various ‘treatment’ milieux. In this paper findings are reported for 70 men and 33 women patients at the Henderson Hospital.

Significantly more patients in the hospital changed in desired directions (‘benefited’) than in previous milieux studied. Grid interpretation of ‘benefit’ seemed to have face value for therapists when compared with clinical impressions.

Association of ‘benefit’ with attendance at meetings is discussed and so are differences in results for men and women. Changes in self‐esteem compare favourably with such changes in the other milieux and for theoretical reasons this suggests that desired changes will be maintained. The hospital did seem to achieve measurable benefit in patients often regarded as unamenable to treatment.
Drop‐out during in‐patient treatment of anorexia nervosa: A clinical study of 133 patients*
Authors: Vandereycken Walter , Pierloot Roland
Published: July 2011

Although anorexia nervosa patients are notorious for their ‘resistance’ to treatment, the phenomenon of drop‐out during in‐patient treatment of these patients is scarcely described in the literature. In a consecutive series of 133 hospitalized female anorexia nervosa patients the authors found a remarkably high drop‐out rate of about 50 per cent, spread over different phases in the therapeutic process. In order to find some ‘risk factors’ concerning the probability of premature discharge against medical advice, some clinical, demographic and follow‐up data were analysed in a retrospective way.

Dropping‐out appears to be a function of age at admission, duration of illness, educational level, social class and treatment method. These findings are interpreted with respect to different types of resistance to psychotherapeutic treatment both on the part of the patient and her family. In most cases a drop‐out has to be considered as a crisis‐related event in the treatment process. It does not only require specific attention, but also reconsideration of the whole therapeutic approach to anorexia nervosa patients.
An association between hostility and poor adherence to treatment in patients suffering from depression
Authors: Pugh Robert
Published: July 2011

Direction of hostility, as measured by the Hostility and Direction of Hostility Questionnaire, was found to be associated with poor compliance during a treatment trial of a new tricyclic antidepressant. An association with other factors such as the type and severity of depression was not found. There are other studies in the literature which support these observations although some controversy surrounds the role played by personality variables in adherence to treatment. Hostility has been shown to be a possible factor in the aetiology of depression, an important factor in the classification of depression, and a symptom of depression which improves with treatment. Hostility may have a particularly large importance in compliance with treatment for depression or may have been overlooked in studies in compliance with treatment for non‐psychiatric disorders.
Transient Thermal Field Measurements in a High Aspect Ratio Channel Related to Transient Thermochromic Liquid Crystal Experiments
Authors: Jenkins Sean C. , Shevchuk Igor V. , von Wolfersdorf Jens , Weigand Bernhard
Published: July 2011

Measurements of transient fluid temperature distributions were made in a high aspect ratio (4:1) internally ribbed two-pass channel relating to the measurement of heat transfer using the transient thermochromic liquid crystal (TLC) technique. The temperature field was measured at several positions leading up to and around the 180 deg bend in a two-passage channel to account for variations in the bulk temperature used as a reference for the transient TLC technique. The results showed that the normalized distribution of the temperature field was time invariant, an important result for the validation of heat transfer results using the transient TLC method. The normalized fluid temperature field was shown to be independent of the inlet temperature step and relatively independent of channel Reynolds number. Fluid temperature distributions were shown to be consistent over the length of the inlet channel; however, temperature field measurements made downstream of the bend exhibited a strong asymmetry. Finally, local temperature distributions were used to adjust the reference temperature used in calculating heat transfer coefficient distributions and to show the behavior of heat transfer due to 180 deg bends.
The Application of Flow Control to an Aft-Loaded Low Pressure Turbine Cascade With Unsteady Wakes
Authors: Bons Jeffrey P. , Pluim Jon , Gompertz Kyle , Bloxham Matthew , Clark John P.
Published: July 2011

The synchronous application of flow control in the presence of unsteady wakes was studied on a highly loaded low pressure turbine blade. At low Reynolds numbers, the blade exhibits a nonreattaching separation bubble under steady flow conditions without upstream wakes. Unsteady wakes from an upstream vane row are simulated with a moving row of bars. The separation zone is modified substantially by the presence of unsteady wakes, producing a smaller separation zone and reducing the area-averaged wake total pressure loss by more than 50%. The wake disturbance accelerates transition in the separated shear layer but stops short of reattaching the flow. Rather, a new time-averaged equilibrium location is established for the separated shear layer. The focus of this study was the application of pulsed flow control using two spanwise rows of discrete vortex generator jets. The jets were located at 59% Cx, approximately the peak cp location, and at 72% Cx. The most effective separation control was achieved at the upstream location. The wake total pressure loss decreased 60% from the wake-only level and the cp distribution fully recovered its high Reynolds number shape. The jet disturbance dominates the dynamics of the separated shear layer, with the wake disturbance assuming a secondary role only. When the pulsed jet actuation was initiated at the downstream location, synchronizing the jet to actuate between wake events was key to producing the most effective separation control. Evidence suggests that flow control using vortex generator jets (VGJs) will be effective in the highly unsteady low pressure turbine environment of an operating gas turbine, provided the VGJ location and amplitude are adapted for the specific blade profile.
Influence of Coolant Density on Turbine Blade Film-Cooling Using Pressure Sensitive Paint Technique
Authors: Narzary Diganta P. , Liu Kuo-Chun , Rallabandi Akhilesh P. , Han Je-Chin
Published: July 2011

Adiabatic film-cooling effectiveness is examined on a high-pressure turbine blade by varying three critical engine parameters, viz., coolant blowing ratio, coolant-to-mainstream density ratio, and freestream turbulence intensity. Three average coolant blowing ratios (BR=1.2, 1.7, and 2.2 on the pressure side and BR=1.1, 1.4, and 1.8 on the suction side), three average coolant density ratios (DR=1.0, 1.5, and 2.5), and two average freestream turbulence intensities (Tu=4.2% and 10.5%) are considered. Conduction-free pressure sensitive paint (PSP) technique is adopted to measure film-cooling effectiveness. Three foreign gases—N2 for low density, CO2 for medium density, and a mixture of SF6 and argon for high density are selected to study the effect of coolant density. The test blade features two rows of cylindrical film-cooling holes on the suction side (45 deg compound), 4 rows on the pressure side (45 deg compound) and 3 around the leading edge (30 deg radial). The inlet and the exit Mach numbers are 0.24 and 0.44, respectively. The Reynolds number of the mainstream flow is 7.5×105 based on the exit velocity and blade chord length. Results suggest that the PSP is a powerful technique capable of producing clear and detailed film-effectiveness contours with diverse foreign gases. Large improvement on the pressure side and moderate improvement on the suction side effectiveness is witnessed when blowing ratio is raised from 1.2 to 1.7 and 1.1 to 1.4, respectively. No major improvement is seen thereafter with the downstream half of the suction side showing drop in effectiveness. The effect of increasing coolant density is to increase effectiveness everywhere on the pressure surface and suction surface except for the small region on the suction side, xss/Cx<0.2. Higher freestream turbulence causes effectiveness to drop everywhere except in the region downstream of the suction side where significant improvement in effectiveness is seen.
Turbine Efficiency for
Unsteady, Periodic Flows
Authors: Suresh Ambady , Hofer Douglas
C. , Tangirala Venkat E.
Published: July 2011

The definition of turbine efficiency for a machine subjected to unsteady periodic inflows is studied. Since mass and energy are conserved quantities over a period, there is no ambiguity in calculating the actual work output of the turbine over a period. The main difficulty lies in calculating the isentropic work output of an ideal turbine operating under the “same” conditions. Two definitions of ideal work output are presented. In the first, the ideal turbine is assumed to operate under the same time traces of inlet and exit total pressures as the actual turbine. The expression for the efficiency that results involves no averages of total pressure. In the second approach, the ideal turbine is assumed to operate under the same average conditions as the actual turbine. Total pressure averages that preserve the isentropic work output are derived and used to calculate an efficiency of the turbine. The two expressions are calculated explicitly for the case of a turbine blade row downstream of a pulse detonation tube. It is found that the definition of efficiency using averages is approximately ten points lower than the first definition.
Three-Dimensional Aerodynamic Optimization for Axial-Flow Compressors Based on the Inverse Design and the Aerodynamic Parameters
Authors: He Liu , Shan Peng
Published: July 2011

Integrating a genetic algorithm code with a response surface methodology code based on the artificial neural network model, this paper develops an optimization system. By introducing a quasi-three-dimensional through-flow design code and a design code of axial compressor airfoils with camber lines of arbitrary shape, and involving a three-dimensional computational fluid dynamics solver, this paper establishes a numerical aerodynamic optimization platform for the three-dimensional blades of axial compressors. The optimization in this paper mainly has four features. First, it applies the conventional inverse design method instead of the common computer aided geometric design parametrization method to generate a three-dimensional blade. Second, it chooses aerodynamic parameters with physical meaning as design variables instead of purely geometrical parameters. Third, it presents a stage-by-stage optimization strategy about the multistage turbomachinery optimization. Fourth, it introduces the visual analysis method into optimization, which can adjust variation ranges of variables by analyzing how great the variables influence the objective function. The above techniques were applied to the redesign of a single rotor row and two double-stage axial fans. The departure angles and work distributions in the inverse design were taken as design variables separately in optimizations of the single rotor and double-stage fans, and they were parametrically represented by means of Bézier curves, whose parameters were used as the optimization variables in the practical operation. The three investigated examples elucidate that not only the techniques mentioned above are appropriate and effective in engineering, but also the design guidance for similar inverse design problems can be obtained from the optimization results.
An Exact Solution for Classic Coupled Thermoelasticity in Cylindrical Coordinates
Authors: Jabbari M. , Dehbani H. , Eslami M. R.
Published: July 2011

In this paper, the classic coupled thermoelasticity model of hollow and solid cylinders under radial-symmetric loading condition (r,t) is considered. A full analytical method is used, and an exact unique solution of the classic coupled equations is presented. The thermal and mechanical boundary conditions, the body force, and the heat source are considered in the most general forms, where no limiting assumption is used.
Strength Behavior of Hot-Tapped Pipeline Under Internal Pressure Load
Authors: Wu B. H. , Sang Z. F. , Wang Z. L. , Widera G. E. O.
Published: July 2011

The objective of this study is to investigate the strength behavior and plastic limit pressure of hot-tapped pipelines under an internal pressure load. Two full scale test models were fabricated for the experiment. A three-dimensional nonlinear finite element analysis was also carried out. The plastic limit pressure was determined approximately by use of the twice the elastic-slope criterion. The results indicate that plastic limit pressures obtained by experiment and finite element analysis are in good agreement. On the basis of the above results, a parametric analysis of the plastic limit pressure was carried out and resulted in the development of formulas useful for the design of hot-tapped pipelines.
Optimum Design of Inhomogeneous Rotating Disks for Minimum Creep Induced Radial Displacement
Authors: Farshi Behrooz , Bidabadi Jalal
Published: July 2011

Gas turbine disks are subject to mechanical stresses due to centrifugal forces exerted by the blades, as well as thermal stresses due to high temperature gradient. High stresses in the presence of elevated temperatures cause the rotating disk material to undergo considerable creep. This phenomenon is significant particularly in cases of turbine blades for power generators, which run almost continuously. Creep strains, in time, lead to deformations resulting in increase of the disk diameter, causing the clearance between blade tips and the turbine’s outer shell to reduce in time. As the above clearance gap is usually limited, this matter is of concern in the design of such equipment for long life. In this investigation, an optimization method is formulated, which is capable of proportioning the thickness distribution of inhomogeneous rotating disks under temperature gradient, so that their long term radii increase due to creep would be minimum. An example is given, which shows the viability of the method.
Optimized growth of Ge nanorod arrays on Si patterns
Authors: Khare C. , Fuhrmann B. , Leipner H. S. , Bauer J. , Rauschenbach B.
Published: July 2011

Self-assembly of polystyrene nanospheres and reactive ion etching have been used to seed Si substrates on which Ge nanorods could be grown by glancing angle deposition (GLAD). This method enables production of large area planar-closed-packed arrays of Ge-GLAD nanostructures on Si seed patterns. A strong column competition on a broad seed width (ws) and a narrow interseed separation distance (Rs) causes the growth of closely bunched multiple structures on the Si seeds. Nanorod growth optimization is realized through the systematic variation of Si seed widths (ws) and the interseed separation distance (Rs), which enable the growth of singular nanorods on each Si seed.
Ultrathin TiSiN overcoat protection layer for magnetic media
Authors: Rose Franck , Marchon Bruno , Rawat Vijay , Pocker Daryl , Xiao Qi-Fan , Iwasaki Tomio
Published: July 2011

TiSiN ultrathin films (10–50 Å) deposited by reactive magnetron sputtering from TixSiy targets were used as anticorrosion overcoats to protect Co-containing recording media. Films’ growth, structure, composition, resistance against hydrolysis, and anticorrosion performances were studied by spectroscopy (x-ray reflectivity and diffraction, ellipsometry, x-ray photoelectron spectroscopy, Fourier transform infrared reflection–absorption spectroscopy) and simulated by molecular dynamics (using modified Tersoff-type interatomic interactions). TiSiN ultrathin overcoats were found to be dense amorphous oxynitrides containing Ti–O–Si linkages. The conversion of SiNx into SiOx by hydrolysis was prevented by introducing less than 20 at. % of Ti in the films. Thanks to the formation of Ti–O–Si linkages which densify the films and reduce oxygen diffusion, good corrosion protection of the magnetic media was achieved down to 28 Å TiSiN overcoat thickness.
On the absence of post-plasma etch surface and line edge roughness in vinylpyridine resists
Authors: Bruce R. L. , Weilnboeck F. , Lin T. , Phaneuf R. J. , Oehrlein G. S. , Long B. K. , Willson C. G. , Alizadeh A.
Published: July 2011

The authors show that poly(4-vinylpyridine) (P4VP) resist eliminates plasma-induced surface roughening for dry etch process conditions (100% Ar, 90% Ar/C4F8) that produce significant roughness in a wide variety of other polymers. In hot-embossed patterned structures, P4VP also shows no sidewall striations and line edge roughness after plasma etching, in contrast to other polymers investigated in this work. The mechanism underlying the retention of smooth surfaces for P4VP was investigated based on the observation that plasma-induced surface roughness in polystyrene (PS) has been linked to wrinkling caused by the formation of a thin, dense, ion-damaged layer. By x-ray photoelectron spectroscopy and in situ ellipsometry analysis, the authors studied two possible mechanisms that would suppress wrinkling in plasma-exposed P4VP: softening of the ion-damaged layer by nitrogen addition and stiffening of the polymer underlayer by VUV modification. While the authors report that the elastic modulus of the ion-damaged layer is reduced in Ar plasma-exposed PS when nitrogen is added to the gas discharge, the ion-damaged layer of P4VP showed no significant change relative to PS. However, by examining only the VUV radiation effect of the Ar discharge on P4VP, evidence of VUV cross-linking was observed. Mechanical stiffening of the VUV cross-linked layer is likely to suppress wrinkling in P4VP when an ion-damaged layer is formed during normal plasma exposure and explain the lack of surface and line edge roughness in the vinylpyridine material.
Chirality in Planar Cell Shape Contributes to Left-Right Asymmetric Epithelial Morphogenesis
Authors: Taniguchi Kiichiro , Maeda Reo , Ando Tadashi , Okumura Takashi , Nakazawa Naotaka , Hatori Ryo , Nakamura Mitsutoshi , Hozumi Shunya , Fujiwara Hiroo , Matsuno Kenji
Published: July 2011

Left-right asymmetry in cell shape is converted to a directional twist of the gut epithelial tube.
Electrical Spiking in
Escherichia coli
Probed with a Fluorescent Voltage-Indicating Protein
Authors: Kralj Joel M. , Hochbaum Daniel R. , Douglass Adam D. , Cohen Adam E.
Published: July 2011

Introducing Bacterial Electrophysiology

Bacterial electrophysiology has been limited by the inability to measure the membrane potential of single cells.

Kralj
et al.

(p.
345
) engineered a class of voltage-sensitive fluorescent membrane proteins to perform electrophysiological measurements on individual intact bacteria. These measurements showed that
Escherichia coli
generate electrical spikes, reminiscent of action potentials in neurons. The response of electrical spiking in bacteria was assessed in response to a wide range of physical and chemical perturbations, and was correlated with efflux activity. In the future, the probe should be useful in determining the roles of membrane potential in a variety of medically, environmentally, and industrially important bacteria.
Control of Mitotic Spindle Angle by the RAS-Regulated ERK1/2 Pathway Determines Lung Tube Shape
Authors: Tang Nan , Marshall Wallace F. , McMahon Martin , Metzger Ross J. , Martin Gail R.
Published: July 2011

During lung development, airway shape is determined by a signaling pathway that regulates the orientation of cell division.
Metallic Quantum Well States in Artificial Structures of Strongly Correlated Oxide
Authors: Yoshimatsu K. , Horiba K. , Kumigashira H. , Yoshida T. , Fujimori A. , Oshima M.
Published: July 2011

Ultrathin films of strontium vanadate display a complex evolution of electronic subbands.
Structure of the FANCI-FANCD2 Complex: Insights into the Fanconi Anemia DNA Repair Pathway
Authors: Joo Woo , Xu Guozhou , Persky Nicole S. , Smogorzewska Agata , Rudge Derek G. , Buzovetsky Olga , Elledge Stephen J. , Pavletich Nikola P.
Published: July 2011

The ID complex reveals how phosphorylation and ubiquitination could influence its stability and DNA-binding functions.
Explosive Percolation Is Continuous
Authors: Riordan Oliver , Warnke Lutz
Published: July 2011

A mathematical proof shows that in many models of the growth of network connectivity, phase transitions are continuous.
Networking—Smoothly Does It
Authors: Janson Svante
Published: July 2011

A rigorous mathematical proof shows that the phase transition in an evolving random network is continuous.
Charging Up Electric Cars
Bottled Lightning
Superbatteries, Electric Cars, and the New Lithium Economy

by Seth Fletcher

Hill and Wang (Farrar, Straus and Giroux), New York, 2011. 272 pp. $26. ISBN 9780809030538.
Authors: Whitacre Jay F.
Published: July 2011

Exploring a crucial aspect of electric-car technologies, Fletcher highlights the hopes for and hurdles facing lithium-ion batteries.
Sympathy for the CEO
Authors: Sapolsky Robert M.
Published: July 2011

Are alpha male baboons more stressed than other high-ranking males?
Tubular Transformations
Authors: Horne-Badovinac Sally , Munro Edwin
Published: July 2011

Two studies offer insight into the cellular processes that produce twists and turns in lung and gut tubes.
Sentence and Word Complexity
Authors: Heinz Jeffrey , Idsardi William
Published: July 2011

Do humans learn the sentence and sound patterns of natural languages through distinct learning mechanisms?
A New Day for Egyptian Science?
Authors: Lawler Andrew
Published: July 2011

In the aftermath of Egypt's popular uprising, the country is embracing a grand vision to make scientific research the engine of a powerhouse economy. Can Egyptian scientists make it happen?
Tackling Egypt's New Plague
Authors: Lawler Andrew
Published: July 2011

Egyptian and Qatari researchers have come up with a nanotechnology method that they say detects hepatitis C RNA more quickly and cheaply than current methods.
Modernizing Mummy Maintenance
Authors: Lawler Andrew
Published: July 2011

At the massive new conservation center of the Grand Egyptian Museum, workers are busy ensuring that the countless beautiful objects entombed in the Giza pyramids will also survive the ages.
Gulf Drilling Disaster Triggers Scrutiny of Mediterranean Oil Rush
Authors: Margottini Laura
Published: July 2011

A rush to find and extract oil in the Mediterranean Sea is threatening one of the planet's marine biodiversity hot spots, scientists warn.
Varying Effects of Different β-Glucans on the Maturation of Porcine Monocyte-Derived Dendritic Cells
Authors: Sonck Eva , Devriendt Bert , Goddeeris Bruno , Cox Eric
Published: July 2011

β-Glucans are well known for their immunomodulatory capacities in humans and mice. For this reason, together with the European ban on growth-promoting antibiotics, β-glucans are intensively used in pig feed. However, as shown in the present study, there is much variation in the stimulatory capacities of β-glucans from different sources. Since dendritic cells (DCs) are the first cells that are encountered after an antigen is taken up by the intestinal epithelial cell barrier, we decided to investigate the effect of two concentrations (5 and 10 μg/ml) of five commercial β-glucan preparations, differing in structure and source, on porcine monocyte-derived dendritic cells (MoDCs). Although all β-glucans gave rise to a significant reduction of the phagocytic activity of DCs, only Macrogard induced a significant phenotypic maturation. In addition to Macrogard, zymosan, another β-glucan derived fromSaccharomyces cerevisiae, and curdlan also significantly improved the T-cell-stimulatory capacity of MoDCs. Most interesting, however, is the cytokine secretion profile of curdlan-stimulated MoDCs, since only curdlan induced significant higher expression levels of interleukin-1β (IL-1β), IL-6, IL-10, and IL-12/IL-23p40. Since the cytokine profile of DCs influences the outcome of the ensuing immune response and thus may prove valuable in intestinal immunity, a careful choice is necessary when β-glucans are used as dietary supplement.
Meningococcal Group C and W135 Immunological Hyporesponsiveness in African Toddlers
Authors: Findlow Helen , Sow Samba , Borrow Ray , Tapia Milagritos , Haidara Fadima Cheick , Akinsola Adebayo K. , Idoko Olubukola T. , Diallo Fatoumata , Adegbola Richard , Tang Yuxiao , Parulekar Varsha , Chadha Helen , Mabey Lesley , Holme Daniel , Townsend Kelly , Chaumont Julie , LaForce F. Marc , Kulkarni Prasad S. , Marchetti Elisa , Viviani Simonetta , Hassan-King Musa , Preziosi Marie-Pierre
Published: July 2011

A phase II clinical study was conducted in African toddlers (aged 12 to 23 months), with subjects receiving either investigational meningococcal group A conjugate (PsA-TT), meningococcal ACWY polysaccharide (PsACWY), orHaemophilus influenzaetype b (Hib-TT) vaccine. Ten months following vaccination, the 3 study groups were further randomized to receive a dose of PsA-TT, a 1/5 dose of PsACWY, or a dose of Hib-TT vaccine. Group A serum bactericidal antibody (SBA) results have been reported previously, with PsA-TT demonstrating superior immunogenicity versus PsACWY vaccine. Immunogenicity for serogroups W135 and C was assessed by SBA assay to investigate the impact of multiple doses in this age group. Blood samples were taken prior to vaccination, 28 days and 40 weeks post-primary vaccination, and 7 and 28 days post-booster vaccination with a 1/5 dose of PsACWY. Subjects who had previously received a full dose of PsACWY had W135 SBA geometric mean titers (GMTs) of 26.1 and 4.4 at 7 and 28 days post-booster vaccination with a 1/5 PsACWY dose, respectively, whereas the W135 SBA GMTs of naïve subjects at these time points following vaccination with a 1/5 dose of PsACWY were 861.1 and 14.6, respectively. Similar differences were observed for serogroup C, with SBA GMTs of 99 and 5.9 at 7 and 28 days post-booster vaccination with a 1/5 dose of PsACWY, respectively, for naïve subjects, compared to 4.1 and 3.2 for previously vaccinated subjects. Immunologic hyporesponsiveness for groups C and W135 was observed following a full dose of PsACWY vaccine at 12 to 23 months of age and a 1/5 dose of PsACWY 10 months later compared to the case for PsACWY-naïve subjects receiving a 1/5 dose of PsACWY vaccine.
Comparison of Plasmid Vaccine Immunization Schedules Using IntradermalIn VivoElectroporation
Authors: Hallengärd David , Haller B. Kristian , Maltais Anna-Karin , Gelius Eva , Nihlmark Kopek , Wahren Britta , Bråve Andreas
Published: July 2011

In vivoelectroporation (EP) has proven to significantly increase plasmid transfection efficiency and to augment immune responses after immunization with plasmids. In this study, we attempted to establish an immunization protocol using intradermal (i.d.) EP. BALB/c mice were immunized with a plasmid encoding HIV-1 p37Gag, either i.d. with the Derma Vax EP device, intramuscularly (i.m.) without EP, or with combinations of both. A novel FluoroSpot assay was used to evaluate the vaccine-specific cellular immune responses. The study showed that i.d. EP immunizations induced stronger immune responses than i.m. immunizations using a larger amount of DNA and that repeated i.d. EP immunizations induced stronger immune responses than i.m. priming followed by i.d. EP boosting. Two and three i.d. EP immunizations induced immune responses of similar magnitude, and a short interval between immunizations was superior to a longer interval in terms of the magnitude of cellular immune responses. The FluoroSpot assay allowed for the quantification of vaccine-specific cells secreting either gamma interferon (IFN-γ), interleukin-2 (IL-2), or both, and the sensitivity of the assay was confirmed with IFN-γ and IL-2 enzyme-linked immunosorbent spot (ELISpot) assays. The data obtained in this study can aid in the design of vaccine protocols using i.d. EP, and the results emphasize the advantages of the FluoroSpot assay over traditional ELISpot assay and intracellular staining for the detection and quantification of bifunctional vaccine-specific immune responses.
Performance of a Rapid and Simple HIV Testing Algorithm in a Multicenter Phase III Microbicide Clinical Trial
Authors: Crucitti Tania , Taylor Doug , Beelaert Greet , Fransen Katrien , Van Damme Lut
Published: July 2011

A multitest sequential algorithm based on rapid and simple (R/S) assays was applied for the diagnosis of HIV infection among participants in a phase 3 microbicide effectiveness trial. HIV testing was performed on finger-prick blood samples obtained from patients after their enrollment in the trial. The specimens were tested in a serial procedure using three different rapid tests (Determine HIV-1/2 [Abbott], SD Bioline HIV-1/2 3.0 [Standard Diagnostics], and Uni-Gold HIV [Trinity Biotech]). In the event of discordant results between the Determine HIV-1/2 and SD Bioline HIV-1/2 3.0 tests, the third assay (Uni-Gold HIV) determined the final outcome. When the final outcome was positive, a second specimen was collected and tested with the same algorithm, only if a positive result was obtained with this sample the participant was informed of her positive serostatus. A total of 5,734 postenrollment specimens obtained from 1,398 women were tested. Forty-six women tested positive according to the testing algorithm performed on the first collected specimen. Confirmatory testing results obtained at the ITM confirmed that 42 women were truly infected. Two of four initial false positives tested negative upon analysis of a second blood specimen. The other two tested false positive twice using specimens collected the same day. A high percentage of specimens reactive with the Determine HIV-1/2 assay was only observed at the study site in Kampala. This result did not appear to be associated with pregnancy or malaria infection. We conclude that HIV testing algorithms, including only R/S assays, are suitable for use in clinical trials, provided that adequate quality assurance procedures are in place.
Persistence of Rabies Antibody 5 Years after Postexposure Prophylaxis with Vero Cell Antirabies Vaccine and Antibody Response to a Single Booster Dose
Authors: Zhang Xiaowei , Zhu Zhenggang , Wang Chuanlin
Published: July 2011

This study was done to investigate the antibody response to a Vero cell antirabies vaccine, the persistence of antibody for 5 years, and the effect of a booster dose after this interval. From August 2005 to February 2011, a total of 195 patients were enrolled into our study due to an animal bite. The Essen intramuscular (i.m.) regimen, which is recommended by the WHO for modern vaccines used in postexposure treatment, was adopted in this study. Blood samples were obtained on day 0, day 7, day 14, day 45, year 1, year 2, year 3, year 4, year 5, and year 5 plus 14 days. Immunogenicity was evaluated by the titration of neutralizing antibodies with a rapid fluorescent focus inhibition test (RFFIT). Seroconversion was expressed as the seroconversion rate (SCR). A secondary quantitative evaluation criterion, other than the seroconversion level, was the geometric mean titer (GMT). Of the 195 enrolled patients, 168 (86.4%) of them completed the whole study. No serious adverse reactions to the vaccine were reported during vaccination, the 5-year follow-up period, or revaccination. On day 14, the rabies antibody GMT value was 8.87 IU/ml in the vaccinees. During the next 5 years, the SCR in the ChengDa vaccine group gradually decreased to 34.0% at year 5, down from 90.5% at year 1. There was a significant booster effect: the GMT was 15.22 IU/ml on year 5 plus 14 days. Our findings demonstrate that the ChengDa rabies vaccine offers an alternative with a high degree of efficacy and yet limited side effects and ensures that the exposed patient will be on the safe side of the risk of rabies by the 14th day. Moreover, when followed by a booster dose 5 years later, it could boost the immunity. A further booster is effective in inducing a good neutralizing antibody response even after an interval of 5 years.
Subcutaneous Immunization with Baculovirus Surface-Displayed Hemagglutinin of Pandemic H1N1 Influenza A Virus Induces Protective Immunity in Mice
Authors: Prabakaran Mookkan , Meng Tao , He Fang , YunRui Tan , Qiang Jia , Lin Raymond Tzer Pin , Kwang Jimmy
Published: July 2011

The protective immunity of baculovirus displaying influenza virus hemagglutinin (BacHA) against influenza 2009 H1N1 virus infection in a murine model was investigated. The results showed that mice vaccinated with live BacHA or an inactive form of adjuvanted BacHA had enhanced specific antibody responses and induced protective immunity against 2009 H1N1 virus infection, suggesting the potential of baculovirus as a live or inactivated vaccine.
Susceptibility of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae According to the New CLSI Breakpoints
Authors: Wang Peng , Hu Fupin , Xiong Zizhong , Ye Xinyu , Zhu Demei , Wang Yun F. , Wang Minggui
Published: July 2011

In 2010 the Clinical and Laboratory Standards Institute (CLSI) lowered the susceptibility breakpoints of some cephalosporins and aztreonam for
Enterobacteriaceae
and eliminated the need to perform screening for extended-spectrum β-lactamases (ESBLs) and confirmatory tests. The aim of this study was to determine how many ESBL-producing strains of three common species of
Enterobacteriaceae
test susceptible using the new breakpoints. As determined with the CLSI screening and confirmatory tests, 382 consecutive ESBL-producing strains were collected at Huashan Hospital between 2007 and 2008, including 158 strains of
Escherichia coli
, 164 of
Klebsiella pneumoniae
, and 60 of
Proteus mirabilis
. Susceptibility was determined by the CLSI agar dilution method. CTX-M-, TEM-, and SHV-specific genes were determined by PCR amplification and sequencing.
bla
CTX-M
genes alone or in combination with
bla
SHV
were present in 92.7% (354/382) of these ESBL-producing strains. Forty-two (25.6%) strains of
K. pneumoniae
harbored SHV-type ESBLs alone or in combination. No TEM ESBLs were found. Utilizing the new breakpoints, all 382 strains were resistant to cefazolin, cefotaxime, and ceftriaxone, while 85.0 to 96.7% of
P. mirabilis
strains tested susceptible to ceftazidime, cefepime, and aztreonam, 41.8 to 45.6% of
E. coli
strains appeared to be susceptible to ceftazidime and cefepime, and 20.1% of
K. pneumoniae
were susceptible to cefepime. In conclusion, all ESBL-producing strains of
Enterobacteriaceae
would be reported to be resistant to cefazolin, cefotaxime, and ceftriaxone by using the new CLSI breakpoints, but a substantial number of ESBL-containing
P. mirabilis
and
E. coli
strains would be reported to be susceptible to ceftazidime, cefepime, and aztreonam, which is likely due to the high prevalence of CTX-M type ESBLs.
Characterization of Emerging GII.g/GII.12 Noroviruses from a Gastroenteritis Outbreak in the United States in 2010
Authors: Takanashi Sayaka , Wang Qiuhong , Chen Ning , Shen Quan , Jung Kwonil , Zhang Zhenwen , Yokoyama Masaru , Lindesmith Lisa C. , Baric Ralph S. , Saif Linda J.
Published: July 2011

Norovirus is a major cause of acute gastroenteritis in humans. A norovirus outbreak occurred in Ohio in January 2010. Stool and saliva samples were obtained from six infected individuals. The full-length genomes of two representative strains (HS206 and HS210) were characterized. They belonged to GII.12 in the capsid but GII.g in the RNA polymerase region. Interestingly, an immunocompetent 2-year-old male shed virus for up to 30 days, as detected by real-time reverse transcription (RT)-PCR. Histo-blood group antigen (HBGA) typing of saliva showed that the norovirus strains infected various types of secretor-positive individuals (types A, B, and O). The viruslike particles of strain HS206 did not bind substantially to A/B/O antigens by synthetic HBGA binding, hemagglutination, or saliva binding assays. These results suggest that infection by this strain may not be A/B/O antigen dependent or thatin vitrobinding patterns do not always accurately reflectin vivoHBGA usage. This is different from the HBGA binding pattern of the previously reported GII.12/Aichi76 strain. Structural analysis of the predicted capsid of these GII.12 strains revealed two amino acid mismatches located near the HBGA binding sites. Four gnotobiotic pigs were inoculated orally with HS206 (6 × 1010genomic equivalents [GE]/pig). Virus shedding began at postinoculation days (PID) 1 to 3 and continued up to PID 16 (1 × 105to 2 × 107GE/ml). Gastroenteritis cases caused by GII.12 noroviruses have been recently reported worldwide. We observed that this emerging GII.12 norovirus infected humans regardless of A/B/O blood type. The infection of pigs by strain HS206 suggests that interspecies transmission of this strain is possible under experimental conditions.
Characterization of Helicobacter pylori cagA andvacAGenotypes among Alaskans and Their Correlation with Clinical Disease
Authors: Miernyk Karen , Morris Julie , Bruden Dana , McMahon Brian , Hurlburt Debby , Sacco Frank , Parkinson Alan , Hennessy Thomas , Bruce Michael
Published: July 2011

Helicobacter pyloriinfection is common in Alaska. The development of severeH. pyloridisease is partially determined by the virulence of the infecting strain. Here we presentvacAandcagAgenotype data forH. pyloristrains isolated from Alaskans and their correlation with clinical disease. We enrolled patients scheduled for esophagogastroduodenoscopy and positive forH. pyloriinfection. Gastric biopsy specimens from the stomach antrum and fundus were cultured. We performed PCR analysis of theH. pylorivacAgene and for the presence of thecagAgene andcagAempty site. We genotyped 515H. pylorisamples from 220 Native and 66 non-Native Alaskans. We detected thecagAgene in 242/286 (85%) persons; of 222 strains that could be subtyped, 95% (212) were non-AsiancagAand 3% (6) were East AsiancagA. After removing mixed infections (n= 17), 83% ofH. pyloristrains had either thevacAs1m1 (120/269) or s2m2 (103/269) genotype. Sixty-six percent (68/103) ofH. pyloristrains with thevacAs2m2 genotype also contained thecagAgene. Infection with anH. pyloristrain having thecagAgene orvacAs1m1 genotype (compared with s1m2 and s2m2) was associated with a decreased risk of esophagitis (P= 0.003 and 0.0003, respectively). Infection with anH. pyloristrain having thevacAs1m1 genotype (compared with s1m2 and s2m2) was associated with an increased risk of peptic ulcer disease (PUD) (P= 0.003). The majority ofH. pyloristrains in this study carried the non-AsiancagAgene and either thevacAs1m1 or s2m2 genotype. A majority ofH. pyloristrains with thevacAs2m2 genotype also contained thecagAgene. There was an association ofH. pylorigenotype with esophagitis and PUD.
Comparison of a Novel Real-Time PCR Assay with Sequence Analysis, Reverse Hybridization, and Multiplex PCR for Hepatitis B Virus Type B and C Genotyping
Authors: Zhao Yao , Zhang Xiu-Yu , Hu Yuan , Zhang Wen-Lu , Hu Jie-Li , Zeng Ai-Zhong , Guo Jin-Jun , Huang Wen-Xiang , Chen Wei-Xian , Shan You-Lan , Huang Ai-Long
Published: July 2011

We compared a novel real-time genotyping and quantitative PCR (GQ-PCR) assay, direct sequence analysis, reverse hybridization, and multiplex PCR for genotyping hepatitis B virus (HBV) in 127 HBV-infected patients. We found that GQ-PCR had the highest concordance with sequence analysis and the highest detection rate for mixed genotype detecting.
Development of a Second Version of the Cobas AmpliPrep/Cobas TaqMan Hepatitis C Virus Quantitative Test with Improved Genotype Inclusivity
Authors: Vermehren Johannes , Colucci Giuseppe , Gohl Peter , Hamdi Nabila , Abdelaziz Ahmed Ihab , Karey Ursula , Thamke Diana , Zitzer Heike , Zeuzem Stefan , Sarrazin Christoph
Published: July 2011

Hepatitis C virus (HCV) RNA measurement has been facilitated by the introduction of real-time PCR-based assays with low limits of detection and broad dynamic ranges for quantification. In the present study, the performance of two second-version prototypes of the Cobas AmpliPrep/Cobas TaqMan HCV Quantitative Test (CAP/CTM v2) with decreased sample input volume and improved genotype inclusivity was investigated. A total of 232 serum and plasma samples derived from patients with chronic hepatitis C (genotype 1 [GT1],
n
= 108; GT2,
n
= 8; GT3,
n
= 24; GT4,
n
= 87; GT5,
n
= 3; and GT6,
n
= 2) were processed in parallel with the Cobas AmpliPrep/Cobas TaqMan HCV Test (CAP/CTM), Cobas Amplicor HCV Monitor Test v2.0 (CAM), and two second-version prototype formulations of CAP/CTM, Mastermix 1 (MMx1) and MMx2. In addition, three GT4 transcripts containing rare variant sequences were tested. The mean log
10
HCV RNA differences for the best-performing CAP/CTM v2/MMx2 formulation in comparison to CAM were −0.05, 0.05, −0.12, −0.10, −0.44, and −0.29 for patients with GT1, GT2, GT3, GT4, GT5, and GT6 infections, respectively. GT1, GT2, and GT4 samples including isolates with known variants within the 5′ untranslated region (G145A, A165T) that were underquantified with CAP/CTM were correctly quantified with the second-version prototype. In addition, CAP/CTM v2 was able to accurately quantify the three transcripts with rare variant sequences. In conclusion, CAP/CTM v2 accurately quantifies HCV RNA across all HCV genotypes, including specimens with rare polymorphisms previously associated with underquantification.
Rapid Discrimination between Candida glabrata, Candida nivariensis, and Candida bracarensis by Use of a Singleplex PCR
Authors: Enache-Angoulvant A. , Guitard J. , Grenouillet F. , Martin T. , Durrens P. , Fairhead C. , Hennequin C.
Published: July 2011

We report here a PCR-based assay using a single primer pair targeting the
RPL31
gene that allows discrimination between
Candida glabrata
,
Candida bracarensis
, and
Candida nivariensis
according to the size of the generated amplicon.
Community Transmission in the United States of a CTX-M-15-Producing Sequence Type ST131 Escherichia coli Strain Resulting in Death
Authors: Owens Robert C. , Johnson James R. , Stogsdill Patricia , Yarmus Lonny , Lolans Karen , Quinn John
Published: July 2011

A middle-aged woman developed fatal urosepsis due to a multidrug-resistant
Escherichia coli
strain representing sequence type ST131, a recently emerged, disseminated, multidrug-resistant extraintestinal pathogen, after presumably having acquired it from her extensively antibiotic-exposed sister with chronic recurrent cystitis. Susceptibility results (reported on day 4) showed resistance to the initially selected regimen.
Identification of Leishmania spp. by Molecular Amplification and DNA Sequencing Analysis of a Fragment of rRNA Internal Transcribed Spacer 2
Authors: de Almeida Marcos E. , Steurer Francis J. , Koru Ozgur , Herwaldt Barbara L. , Pieniazek Norman J. , da Silva Alexandre J.
Published: July 2011

Isoenzyme analysis of cultured parasites is the conventional approach for
Leishmania
species identification. Molecular approaches have the potential to be more sensitive and rapid. We designed PCR generic primers to amplify a segment of the rRNA internal transcribed spacer 2 (ITS2) from multiple
Leishmania
species. To validate the selected ITS2 fragment, we tested clinical specimens and compared the species results obtained by the molecular approach (PCR followed by DNA sequencing analysis) with those from the parasitologic approach (
in vitro
culture followed by isoenzyme analysis). Among the 159 patients with clinical specimens positive by both approaches, a total of eight
Leishmania
species were identified. The species results were concordant for all but two patients: for one patient, the results were
Leishmania
(
Viannia
)
guyanensis
by the molecular approach versus
L. (V.) braziliensis
by the parasitologic approach; for the other patient, the results were
L.
(
Leishmania
)
tropica
versus
L. (L.) major
, respectively. ITS2 PCR, followed by sequencing analysis, can be used to detect and discriminate among
Leishmania
species. The results confirmed our hypothesis that a region of the ITS2 gene can complement the characterization of
Leishmania
parasites at the species level. The approach we developed can be used as a diagnostic tool in reference laboratories with adequate infrastructure to perform molecular characterization of pathogens.
Multiplex Real-Time PCR Melting Curve Assay To Detect Drug-Resistant Mutations of Mycobacterium tuberculosis
Authors: Luo Tao , Jiang Lili , Sun Weiming , Fu G. , Mei Jian , Gao Qian
Published: July 2011

Early diagnosis of drug-resistant
Mycobacterium tuberculosis
is urgently needed to optimize treatment regimens and to prevent the transmission of resistant strains. Real-time PCR assays have been developed to detect drug resistance rapidly, but none of them have been widely applied due to their complexity, high cost, or requirement for advanced instruments. In this study, we developed a real-time PCR method based on melting curve analysis of dually labeled probes. Six probes targeting the
rpoB
81-bp core region,
katG315
, the
inhA
promoter, the
ahpC
promoter, and
embB306
were designed and validated with clinical isolates. First, 10 multidrug-resistant (MDR) strains with a wide mutation spectrum were used to analyze the melting temperature (

T
m

) deviations of different mutations by single real-time PCR. All mutations can be detected by significant

T
m

reductions compared to the wild type. Then, three duplex real-time PCRs, with two probes in each, were developed to detect mutations in 158 MDR isolates. Comparison of the results with the sequencing data showed that all mutations covered by the six probes were detected with 100% sensitivity and 100% specificity. Our method provided a new way to rapidly detect drug-resistant mutations in
M. tuberculosis.
Compared to other real-time PCR methods, we use fewer probes, which are labeled with the same fluorophore, guaranteeing that this assay can be used for detection in a single fluorescent channel or can be run on single-channel instruments. In conclusion, we have developed a widely applicable real-time PCR assay to detect drug-resistant mutations in
M. tuberculosis
.