bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of three diagnostic techniques for detection of rotavirus and coronavirus in calf faeces in Australia
Authors: Izzo MM , Kirkland PD , Gu X , Lele Y , Gunn AA , House JK
Published: March 2012

Objective  Compare real‐time reverse transcription polymerase chain reaction (qRT‐PCR), a commercially available enzyme‐linked immunosorbent assay (ELISA) and lateral flow immunochromatography assay (LAT) for the detection of rotavirus and coronavirus in faecal samples collected from diarrhoeic calves.

Design  Prospective survey.

Method  Samples were tested at two separate facilities using a commercial ELISA and an in‐house qRT‐PCR. Simple logistic regression was performed to examine the relationship between the two tests. A subset of samples was screened using qRT‐PCR, ELISA and a commercial LAT dipstick (132 faecal samples were tested for coronavirus and 122 samples for rotavirus).

Results  Of the 586 samples tested, 131 (22.39%) and 468 (79.86%) were positive for coronavirus and group A rotavirus, respectively, using qRT‐PCR. The number of samples positive on ELISA for coronavirus and rotavirus was 73 (12.46%) and 225 (38.40%), respectively. Using LAT, 30 (22.73%) and 43 (35.35%) samples were positive for coronavirus and rotavirus, respectively. Simple linear regression revealed a statistically significant (P < 0.05) but weak (r2=−0.07 and −0.40) correlation between the rotavirus/coronavirus qRT‐PCR and ELISA, respectively. There was also poor agreement between the LAT and qRT‐PCR assays.

Conclusion  The sensitivity and specificity of the commercial ELISA and LAT assays evaluated in this study were low compared with qRT‐PCR. The low positive and negative predictive values of the assays suggests that they were of limited diagnostic benefit in the population sampled.
Intravesical Application of Lidocaine and Sodium Bicarbonate in the Treatment of Obstructive Idiopathic Lower Urinary Tract Disease in Cats
Authors: Zezza L. , Reusch C.E. , Gerber B.
Published: March 2012

Background

In human patients with interstitial cystitis, intravesical instillation of alkalinized lidocaine sometimes is associated with sustained amelioration of symptoms beyond the acute treatment phase. Interstitial cystitis shares many features in common with feline idiopathic cystitis.

Objective

To evaluate whether intravesical instillation of alkalinized lidocaine decreases recurrence of urethral obstruction and severity of clinical signs in cats with obstructive idiopathic LUTD.

Animals

Twenty‐six cats with obstructive idiopathic LUTD. Twelve cats in case group (treatment with alkalinized lidocaine) and 14 control cats (treatment with placebo or standard treatment).

Methods

Cats were randomly assigned to treatment (2 or 4 mg/kg lidocaine and sodium bicarbonate) or placebo groups (0.2 mL/kg saline solution and sodium bicarbonate). The intravesical instillation was done once a day for 3 days. Some cats underwent standard treatment only (indwelling urinary catheter for 3 days without intravesical instillations). A 2‐week, 1‐month, and 2‐month follow‐up after treatment was made using a questionnaire. The recurrence rate and amelioration scores of clinical signs were assessed and compared.

Results

Recurrence of urethral obstruction was 58% (7/12) in the case group and 57% (8/14) in the control group. Amelioration scores were similar between the 2 groups.

Conclusion and Clinical Importance

Intravesical administration of lidocaine for up to 3 consecutive days had no apparent beneficial effect on decreasing recurrence rate and severity of clinical signs in cats with obstructive idiopathic LUTD.
Dynamic respiratory endoscopy of Standardbred racehorses during qualifying races
Authors: PRIEST D. T. , CHEETHAM J. , REGNER A. L. , MITCHELL L. , SODERHOLM L. V. , TAMZALI Y. , DUCHARME N. G.
Published: March 2012

Reason for performing study:Examination of the equine upper airway during racing has not previously been documented.

Objective:To describe the feasibility and appearance of the upper airways by overground respiratory endoscopic examination during racing conditions.

Methods:Overground videoendoscopic examinations were performed on 46 Standardbred racehorses during qualifying races. Examined horses' speeds were recorded throughout the race with a portable GPS device.

Results:The procedure did not interfere with performance as there were no significant differences in race times between races in which horses were examined with the endoscope in place and prior unexamined races. Airway obstructions during or after the race were documented in 21 horses. Most previously reported causes of upper airway obstruction were observed; surprisingly bilateral ventro‐medial arytenoid displacement (VMAD; n = 5) was seen during exercise as frequently as dorsal displacement of the soft palate (DDSP). Although DDSP (n = 10) was the most common diagnosis made, many displacements (n = 5) occurred after the race. Horses that demonstrated DDSP after the race had slower speeds than unaffected horses during the race.

Conclusions:Racing endoscopy permits the diagnosis of upper airway obstructions without affecting performance. The occurrence of DDSP immediately after exercise may be clinically relevant. During racing VMAD may be an important anomaly.

Potential relevance:Racing endoscopy could be used to correlate the sensitivity of diagnostic endoscopy during race‐training or treadmill examination. The pathogenesis and significance of VMAD deserves further investigation.
Enhancement of gene expression and melanin production of human tyrosinase gene loaded in elastic cationic niosomes
Authors: Khositsuntiwong Narinthorn , Manosroi Aranya , Götz Friedrich , Werner Rolf G , Manosroi Worapaka , Manosroi Jiradej
Published: March 2012

Objectives

Disturbance in the synthesis of tyrosinase might be one of the major causes of vitiligo. The enhancement of tyrosinase gene expression and melanin production by loading the plasmid in elastic cationic niosomes was investigated in tyrosinase gene knocked out human melanoma (M5) cells and in tyrosine-producing mouse melanoma (B16F10) cells.

Methods

Niosomes composed of Tween 61/cholesterol/dimethyl dioctadecyl ammonium bromide at 1 : 1 : 0.5 molar ratio were prepared by the freeze-dried empty liposomes method. The thin lipid film was redissolved in distilled water or 25% ethanol to obtain the non-elastic or elastic cationic niosomes, respectively.

Key findings

The maximum loading of the plasmid in non-elastic and elastic niosomes was 130 and 100 µg per 16 mg of the niosomal contents, respectively. The plasmid-loaded elastic cationic niosomes exhibited high specific tyrosinase activity of 1.66 and 1.50 fold in M5 cells and 6.81 and 4.37 fold in B16F10 cells compared with the free plasmid and the plasmid-loaded non-elastic cationic niosomes, respectively.

Conclusions

This study has demonstrated not only the enhancement of the expression of human tyrosinase gene by loading in elastic cationic niosomes, but also the potential application of this gene delivery system for the further development of vitiligo gene therapy.
Virtual Clay Modeling System Based on Aesthetic Curves
Authors: Aoyama Hideki , Kai Hiroshige
Published: March 2012

Reverse engineering is applied to design sophisticated forms such as car body. In the design process of reverse engineering, a clay model is created to express the designer’s idea and to evaluate the designed shape, after which a CAD model is constructed from the measurement data of the clay model. Modeling a shape in clay takes considerable time and costs, so there are strong demands for a more efficient process. In this study, a design support system which enables designers to model shapes in virtual clay in virtual space was developed. Since the process is carried out using digital data, the time and costs for the modeling are drastically reduced. In the modeling process, virtual clay is cut and deformed using digital tools with aesthetic curves which can be easily and intuitively defined by the designer. In order to define aesthetic curves, two methods are proposed; one by using the deflection curves of a plate spring with minimum strain energy and the other by using constant rhythm curves with constant curvature changing rate. Modeling tests were also carried out to verify the validity of the developed system.
An Immersive Virtual Environment for Varying Risk and Immersion for Effective Training
Authors: Harter Derek , Lu Shulan , Kotturu Pratyush , Pierce Devin
Published: March 2012

We present an immersive virtual environment being developed to study questions of risk perception and their impacts on effective training. Immersion is known to effect the quality of training in virtual environments, and the successful transfer of skills to real world situations. However, the level of perceived immersiveness that an environment invokes is an ill defined concept, and effects of different types of immersion are known to have greater and lesser influences on training outcomes. We concentrate on how immersiveness effects perceived risk in virtual environments, and how risk impacts training effectiveness. Simulated risk can invoke an alief of danger in subjects using a virtual environment. Alief is a concept useful in virtual training that describes situations where the person experiencing a simulated scenario knows it is not real, but suspends disbelief (willingly or unwillingly). This suspension of belief (alief) can cause the person to experience the same sorts of autonomic reactions as if they were experiencing the situation in real life (for example, think of fear invoked on amusement park rides). Alief of risk or danger has been proposed as one phenomenon that can influence training outcomes, like the experience of immersion, when training in virtual environments. In this paper we present work on developing a low-cost virtual environment for the manipulation of immersion and risk for cognitive studies. In this environment we provide several alternative input modalities, from mouse to Wii remote interactivity, to control a virtual avatar’s hand and arm for performing risky every day tasks. Immersion can be manipulated in several ways, as well as the type and risk associated with tasks. Typical tasks include performing kitchen preparation work (using knives or hot items), or wood or metal working tasks (involving manipulation of dangerous tools). This paper describes the development and technologies used to create the virtual environment, and how we vary risk perception and immersion of users for various cognitive tasks. The capabilities and manipulations of immersiveness and risk are presented together with some findings on using Wii motes as input devices in several ways for virtual environments. The paper concludes with some preliminary results of varying perceived risk on cognitive task performance in the developed environment.
Survey on Virtual Prototyping Technologies for Orthopedic Implants and Prosthesis Design
Authors: Bandera Camillo , Filippi Stefano , Motyl Barbara
Published: March 2012

Nowadays, Virtual Prototyping (VP) methods are widely used for product design and development purposes. In particular, VP methods are now used also for the orthopedic products development process, to better understand the functional performance of prostheses or implants within the musculoskeletal system. In fact, developing validated virtual models of joints or of other anatomical structures may reduce design and prototyping costs and compress development cycles. The purpose of this paper is to point out first the state of art of both the VP technologies and the kinds of virtual models used in the medical field, with particular attention to those used for the design and development of orthopedic products. Then, it focuses on a qualitative analysis of some biomechanical simulation software packages (LifeMOD, AnyBody, and OpenSim) as tools for the improvement of the product design and development processes.
The Role of High Visual Realism in Reducing Potential Risk Taking in Simulated Environments
Authors: Lu Shulan , Pierce Devin , Rawlinson Terry , Harter Derek
Published: March 2012

Virtual environments (VEs) are developed to invoke feelings of presence in the digitally created representations, which leads to people perceiving and enacting actions as they would in corresponding real world environments. Even though significant strides have been made in enhancing the level of realism of virtual systems, there is still a long way to go toward a system that could provide full immersive experiences. Furthermore, the development cycle of a high realism system can be time consuming and costly. On the theoretical side, this desire of achieving the feeling of presence is not always consistent with the body of literature on grounded cognition, where the environment is known to significantly impact the user’s perception and action. Nevertheless, many studies have shown that people reported the feeling of presence even though the VEs they interacted with were far from realistic representations of the actual environments. This raised a question as to what dimensions of the environments are essential in triggering the feelings of presence. Instead of taking the approach where a fairly immersive system is used and a particular variable is investigated to ascertain its effects on the feelings of presence, the approach used in the current work investigated whether people reduce the potential injury to an avatar by starting out with a low to medium visual realism environment and scaling up to a higher visual realism. The results in the current study suggest that compared to enacting actions in the real world with one’s own body, people are more likely to bring injury to an avatar. This indicates that high visual realism may not be the essential ingredient in invoking the feelings of presence that regulate risk taking behaviors. The limitation and the next step of this research are discussed.
Virtual Reality Application in Case New Holland Product Development
Authors: Monacelli Gennaro , Li Kezhun , Brokaw Andrew
Published: March 2012

This paper describes the VR technologies Case New Holland has implemented in the past few years and their impact on product development by use various examples. It has shown that VR can be used through the entire product life cycle with significant benefits. It enables developmental personnel through the entire enterprise to get involved much earlier and engaged in a much more proactive way. It also shows that developing a process that is efficient and seamless through development cycle is critical to VR’s success.
Study of the Influence of Different Washout Algorithms on Simulator Sickness for a Driving Simulation Task
Authors: Aykent Barıs¸ , Paillot Damien , Me´rienne Fre´de´ric , Fang Zhou , Kemeny Andras
Published: March 2012

This paper deals with the effects of different washout algorithms used for Stewart platforms on subjective and objective ratings. Washout algorithms are used to represent vehicle dynamics in a restricted spatial place. An adaptive washout algorithm was realized to control the hexapod platform, depending on the specific force error in longitudinal, lateral and vertical directions, in order to compare user’s experience with those in the case of classical algorithm. In this study, the simulator sickness has been evaluated for three algorithms in dynamic driving simulator situation in objective and subjective way.
Virtual Prototyping Technique Applied to the Design of a Process Reciprocating Compressor
Authors: Traversari Riccardo , Rossi Alessandro , Faretra Marco
Published: March 2012

Virtual Prototyping (VP) reproduces a complete machine to test it several times, as a scale 1:1 laboratory prototype. VP utilizes various CAE tools, such as 3D modelling, Structural FEA, Multibody Dynamic Analysis (MDA), Multiaxial fatigue analysis, and Fluid Structure Interaction (FSI) in an integrated way. The VP of a rotating machine allows considering a realistic stepless loading pattern throughout the complete revolution and determining automatically the fatigue safety factors within the whole machine structural assembly, while FSI allows dealing simultaneously with thermodynamic, motion and deformation phenomena. This approach was used to review the design of the crank mechanism and cylinders of an existing reciprocating compressor. The loads (including inertia forces) were applied to the gudgeon pin and, by means of the MDA, to all the other components. An advanced approach, based on Fluid Structure Interaction (FSI) analysis, was applied for the thermodynamic analysis of the cylinder’s efficiency. A 3D CFD Model, simulating the cylinder with mobile piston and valves, was developed and experimentally validated. The 3D domain simulating the compression chamber changes with the piston motion law while valve rings move according to the fluid dynamic forces.
Table Top Augmented Reality System for Conceptual Design and Prototyping
Authors: VanWaardhuizen Michael , Oliver James , Gimeno Jesus
Published: March 2012

The AugmenTable is a desktop augmented reality workstation intended for conceptual design and prototyping. It combines a thin form factor display, inexpensive web cameras, and a PC into a unique system that enables natural interaction with virtual and physical parts. This initial implementation of the AugmenTable takes advantage of the popular open source augmented reality software platform ARToolkit to enable manual interaction with physical parts, as well as interaction with virtual parts via a physically marked pointer or a color-marked fingertip. This paper describes similar previous work, the methods used to create the AugmenTable, the novel interaction it affords users, and a number of avenues for advancing the system in the future.
Developing the Planck Mission Simulation as a Multi-Platform Immersive Application
Authors: Dekker Gerald A. , Moreland John , van der Veen Jatila
Published: March 2012

Planck is an international mission led by the European Space Agency with significant contribution by NASA, designed to measure the anisotropy of the Cosmic Microwave Background (CMB), the oldest radiation of the universe, with the greatest accuracy and precision of any such CMB experiment to date. The present work was completed as part of the Planck Education and Public Outreach (E/PO) effort to communicate the results of Planck science to the public. The Planck Mission Simulation is a multiplatform, interactive visualization of the mission, from launch to orbital insertion to data gathering operations. The simulation was developed for a number of hardware and software configurations. Originally designed for a multi-screen virtual reality system, the scope of project grew to include other systems, including 3D kiosk displays, stereoscopic televisions, and domed-roomed systems. Implementation factors, technical details, and lessons learned from deployment on various platforms are discussed.
Enhancement of the SAAM Driving Simulator Graphics Pipeline for Speed Perception Studies
Authors: Colombet Florent , Paillot Damien , Me´rienne Fre´de´ric , Kemeny Andras
Published: March 2012

This paper presents a speed perception study focusing on the impact of a visual scale factor. This factor corresponds to the ratio between the geometric field of view (of the camera) and the driver’s field of view covered by the screen. 20 participants have reproduced 2 speeds (50 and 90 km/h) without knowing the numerical values of these consigns, and with 5 different visual scale factors: 0.70, 0.85, 1.00, 1.15 and 1.30. We show that this visual factor has a significant impact on the speed reached by the subjects and that the variation of perceived speed can be deducted from the used visual scale factor. To be able to conduct this experiment, we had first to solve an image correction issue. As the visual display is projected on a cylindrical screen, a distortion correction must be performed. This correction is dependent on the viewer’s position and so is also dependent on the visual scale factor. We had then to improve our image warping approach to be able to take into account in real time the observer’s position (which was not the case until now). This paper presents a concise state of the art about image correction and some details about the implementation of the chosen algorithm.
Configuring Virtual Reality Displays in a Mixed-Reality Environment for LVC Training
Authors: Newendorp Brandon J. , Noon Christian , Holub Joe , Winer Eliot H. , Gilbert Stephen , de la Cruz Julio
Published: March 2012

In order to adapt to an ever-changing set of threats, military forces need to find new methods of training. The prevalence of commercial game engines combined with virtual reality (VR) and mixed reality environments can prove beneficial to training. Live, virtual and constructive (LVC) training combines live people, virtual environments and simulated actors to create a better training environment. However, integrating virtual reality displays, software simulations and artificial weapons into a mixed reality environment poses numerous challenges. A mixed reality environment known as The Veldt was constructed to research these challenges. The Veldt consists of numerous independent displays, along with movable walls, doors and windows. This allows The Veldt to simulate numerous training scenarios. Several challenges were encountered in creating this system. Displays were precisely located using the tracking system, then configured using VR Juggler. The ideal viewpoint for each display was configured based on the expect location for users to be looking at it. Finally, the displays were accurately aligned to the virtual terrain model. This paper describes how the displays were configured in The Veldt, as well as how it was used for two training scenarios.
Usefulness of Virtual 3D Modeling to Visualize the Effect of Uncertain Data in Groundwater Solute Transport
Authors: Viswanathan Chandramouli , Zhou Chenn , Moreland John , Guo Site
Published: March 2012

Immersive virtual reality models can be effectively used in understanding the system behavior. It will also help the students to understand the influences of boundary conditions by reflection. In particular, those systems which are not suitable for laboratory experiment can be handled using virtual reality platform. In this research, groundwater solute transport simulation was developed as a laboratory module. Groundwater system movement takes place beneath ground. Recently, several sites were contaminated due to manmade activities. This complex process is very difficult to understand with varying boundary conditions. If there is an uncertainity associated with the modeling parameters, remedial alternatives planned may be not as effective as expected. Behavior changes under these circumstances can be tackled through a sensitivity analysis and incorporated in virtual 3D. In this study, by varying the hydraulic conductivity systematically in different layers, the possible variations in the groundwater contamination plume movements were examined for different simulation time domains with remedial alternatives.
Expanding Haptic Workspace for Coupled-Object Manipulation
Authors: Pavlik Ryan A. , Vance Judy M.
Published: March 2012

Haptic force-feedback offers a valuable cue in exploration and manipulation of virtual environments. However, grounding of many commercial kinesthetic haptic devices limits the workspace accessible using a purely position-control scheme. The bubble technique has been recently presented as a method for expanding the user’s haptic workspace. The bubble technique is a hybrid position-rate control system in which a volume, or “bubble,” is defined entirely within the physical workspace of the haptic device. When the device’s end effector is within this bubble, interaction is through position control. When exiting this volume, an elastic restoring force is rendered, and a rate is applied that moves the virtual accessible workspace. Existing work on the bubble technique focuses on point-based touching tasks. When the bubble technique is applied to simulations where the user is grasping virtual objects with part-part collision detection, unforeseen interaction problems surface. This paper discusses three details of the user experience of coupled-object manipulation with the bubble technique. A few preliminary methods of addressing these interaction challenges are introduced.
Ultra-high vacuum deposition and characterization of silicon nitride thin films
Authors: Katzer D. S. , Meyer D. J. , Storm D. F. , Mittereder J. A. , Bermudez V. M. , Cheng S. F. , Jernigan G. G. , Binari S. C.
Published: March 2012

Silicon nitride thin films were deposited on (100) Si wafers in an ultra-high vacuum system using a Si effusion cell and reactive nitrogen from a radio-frequency plasma source. The films were characterized using infrared transmission spectroscopy, infrared reflectance, Rutherford backscattering spectrometry, spectroscopic ellipsometry, specular x-ray reflectivity, wet etching in a buffered-oxide etch solution, and the electrical characterization of metal-insulator-semiconductor capacitors. High-quality, stoichiometric silicon nitride films with a refractive index of 2.05 at 632.8 nm are produced when the deposition temperature is 750 °C. Lower deposition temperatures produce nitrogen-rich silicon nitride films with lower refractive index, lower density, greater tendency toward oxidation in ambient air, faster etching in a buffered oxide etch solution, and greater electrical leakage. A deposition model involving thermal evolution of weakly-bonded excess N is proposed to explain our observations.
Resistive switching of HfO2 based flexible memories fabricated by low temperature atomic layer deposition
Authors: Fang R. C. , Wang L. H. , Yang W. , Sun Q. Q. , Zhou P. , Wang P. F. , Ding S. J. , Zhang David W.
Published: March 2012

HfO2-based flexible memories were fabricated using a low temperature atomic layer deposition (LTALD) process to examine resistive switching performance. The devices exhibit typical bipolar resistive switching. The endurance and retention behaviors were also investigated. No significant degradation of the device was noted at either room temperature or 85 °C, and the current transport mechanism of the high- and low-resistance states are estimated to be Ohmic and trap-assisted current, respectively. The authors propose that this LTALD process will significantly improve fabrication of flexible memories.
Monte Carlo simulation of breast imaging using synchrotron radiation
Authors: Fitousi N. T. , Delis H. , Panayiotakis G.
Published: March 2012

Purpose:

Synchrotron radiation (SR), being the brightest artificial source of x‐rays with a very promising geometry, has raised the scientific expectations that it could be used for breast imaging with optimized results. The “in situ” evaluation of this technique is difficult to perform, mostly due to the limited available SR facilities worldwide. In this study, a simulation model for SR breast imaging was developed, based on Monte Carlo simulation techniques, and validated using data acquired in the SYRMEP beamline of the Elettra facility in Trieste, Italy. Furthermore, primary results concerning the performance of SR were derived.

Methods:

The developed model includes the exact setup of the SR beamline, considering that the x‐ray source is located at almost 23 m from the slit, while the photon energy was considered to originate from a very narrow Gaussian spectrum. Breast phantoms, made of Perspex and filled with air cavities, were irradiated with energies in the range of 16–28 keV. The model included a Gd2O2S detector with the same characteristics as the one available in the SYRMEP beamline. Following the development and validation of the model, experiments were performed in order to evaluate the contrast resolution of SR. A phantom made of adipose tissue and filled with inhomogeneities of several compositions and sizes was designed and utilized to simulate the irradiation under conventional mammography and SR conditions.

Results:

The validation results of the model showed an excellent agreement with the experimental data, with the correlation for contrast being 0.996. Significant differences only appeared at the edges of the phantom, where phase effects occur. The initial evaluation experiments revealed that SR shows very good performance in terms of the image quality indices utilized, namely subject contrast and contrast to noise ratio. The response of subject contrast to energy is monotonic; however, this does not stand for contrast to noise ratio, since there is a range of optimal performance for SR (18–21 keV). In comparison to conventional mammography, SR shows improved subject contrast for energies lower than the mean energy of each spectrum.

Conclusions:

The comparison of the results of the two models, conventional and SR, proved that SR exhibits better performance in the majority of cases. The proposed simulation model offers the possibility to perform exhaustive search to evaluate the performance of SR in clinical applications such as breast imaging.
Prostaglandin D
2
Inhibits Hair Growth and Is Elevated in Bald Scalp of Men with Androgenetic Alopecia
Authors: Garza Luis A. , Liu Yaping , Yang Zaixin , Alagesan Brinda , Lawson John A. , Norberg Scott M. , Loy Dorothy E. , Zhao Tailun , Blatt Hanz B. , Stanton David C. , Carrasco Lee , Ahluwalia Gurpreet , Fischer Susan M. , FitzGerald Garret A. , Cotsarelis George
Published: March 2012

Prostaglandin D
2
inhibits hair growth through its receptor, GPR44, and this pathway could serve as a new target for developing treatments for male pattern baldness.
Merging Systems Biology with Pharmacodynamics
Authors: Iyengar Ravi , Zhao Shan , Chung Seung-Wook , Mager Donald E. , Gallo James M.
Published: March 2012

Enhanced pharmacodynamic models combine favorable features of systems biology and traditional models and may form the basis of precision medicine.
The Gall of Some Bacteria: Strange Behavior by
Salmonella
Authors: Shrout Joshua D.
Published: March 2012

Salmonella
shows reduced virulence gene expression when growing on bile in the gallbladder.
Modulation of Anti-Tumor Necrosis Factor Alpha (TNF-α) Antibody Secretion in Mice Immunized with TNF-α Kinoid
Authors: Assier Eric , Semerano Luca , Duvallet Emilie , Delavallée Laure , Bernier Emilie , Laborie Marion , Grouard-Vogel Géraldine , Larcier Patrick , Bessis Natacha , Boissier Marie-Christophe
Published: March 2012

Tumor necrosis factor alpha (TNF-α) blockade is an effective treatment for patients with TNF-α-dependent chronic inflammatory diseases, such as rheumatoid arthritis, Crohn's disease, and psoriasis. TNF-α kinoid, a heterocomplex of human TNF-α and keyhole limpet hemocyanin (KLH) (TNF-K), is an active immunotherapy targeting TNF-α. Since the TNF-K approach is an active immunization, and patients receiving this therapy also receive immunosuppressant treatment, we evaluated the effect of some immunosuppressive drugs on the generation of anti-TNF-α antibodies produced during TNF-K treatment. BALB/c mice were injected intramuscularly with TNF-K in ISA 51 adjuvant. Mice were also injected intraperitoneally with one of the following: phosphate-buffered saline, cyclophosphamide, methylprednisolone, or methotrexate. Anti-TNF-α and anti-KLH antibody levels were assessed by enzyme-linked immunosorbent assay and the anti-TNF-α neutralizing capacity of sera by L929 bioassay. Our results showed that current treatments used in rheumatoid arthritis, such as methylprednisolone and methotrexate, do not significantly alter anti-TNF-α antibody production after TNF-K immunization. In contrast, the administration of cyclophosphamide (200 mg/kg) after immunization significantly reduced anti-TNF-α antibody titers and their neutralizing capacity.
Molecular Detection of Capillaria aerophila, an Agent of Canine and Feline Pulmonary Capillariosis
Authors: Di Cesare Angela , Castagna Giuseppe , Otranto Domenico , Meloni Silvana , Milillo Piermarino , Latrofa Maria Stefania , Paoletti Barbara , Bartolini Roberto , Traversa Donato
Published: March 2012

Capillaria aerophila
, a trichuroid nematode causing pulmonary infections in wild and domestic carnivores, is occasionally and potentially poorly recognized in infections of humans due to clinicopathological mimicry and a lack of accurate, robust laboratory diagnostics. The present work evaluated the efficiency of a DNA-based assay amplifying a partial cytochrome
c
oxidase subunit 1 (
cox
1) gene of
C. aerophila
in the diagnosis of lung capillariosis. Fecal samples from 34 dogs and 10 cats positive at parasitological examination for
C. aerophila
and other endoparasites (i.e., other lungworms, whipworms, roundworms, hookworms, tapeworms, and/or coccidia) and from 44 animals negative for
C. aerophila
but positive for other endoparasites were molecularly examined. Of the 44 samples positive for
C. aerophila
at copromicroscopy, 43 scored positive (i.e., 33/34 dogs and 10/10 cats) in seminested PCR, resulting in a sensitivity of 97 to 100%. Samples that were copromicroscopy negative for
C. aerophila
although positive for other endoparasites never produced a PCR product or nonspecific amplicons. The specific PCR amplification of
C. aerophila
(i.e., specificity of 100%) was confirmed by a nucleotide sequence analysis of the
cox
1 amplicons. The potential implications of the molecular diagnosis of lung capillariosis are discussed.
Cost Savings Associated with Testing of Antibodies, Antigens, and Nucleic Acids for Diagnosis of Acute HIV Infection
Authors: Karris Maile Y. , Anderson Christy M. , Morris Sheldon R. , Smith Davey M. , Little Susan J.
Published: March 2012

Efforts to identify all persons infected with HIV in the United States are driven by the hope that early diagnosis will lower risk behaviors and decrease HIV transmission. Identification of HIV-infected people earlier in the course of their infection with HIV antigen/antibody (Ag/Ab) combination assays (4th-generation HIV assays) should help achieve this goal. We compared HIV RNA nucleic acid test (NAT) results to the results of a 4th-generation Ag/Ab assay (Architect HIV Ag/Ab Combo [HIV Combo] assay; Abbott Diagnostics) in 2,744 HIV antibody-negative samples. Fourteen people with acute HIV infection (HIV antibody negative/NAT positive) were identified; the HIV Combo assay detected nine of these individuals and was falsely negative in the remaining five. All five persons missed by the HIV Combo assay were in the stage of exponential increase in plasma virus associated with acute HIV infection (3, 7, 20, 35, 48). In contrast, most acutely infected persons detected by the HIV Combo assay demonstrated either a plateauing or decreasing plasma viral load. The HIV Combo assay also classified as positive five other samples which were negative by NAT. Taken together, the HIV Combo assay had a sensitivity of 73.7% and a specificity of 99.8%. Using published data, we estimated secondary transmission events had HIV infection in these five individuals remained undiagnosed. Screening of our population with NAT cost more than screening with the HIV Combo assay but achieved new diagnoses that we predict resulted in health care savings that far exceed screening costs. These findings support the use of more sensitive assays, like NAT, in HIV screening of populations with a high prevalence of acute HIV infection.
Novel Molecular Method for Identification of Streptococcus pneumoniae Applicable to Clinical Microbiology and 16S rRNA Sequence-Based Microbiome Studies
Authors: Scholz Christian F. P. , Poulsen Knud , Kilian Mogens
Published: March 2012

The close phylogenetic relationship of the important pathogen
Streptococcus pneumoniae
and several species of commensal streptococci, particularly
Streptococcus mitis
and
Streptococcus pseudopneumoniae
, and the recently demonstrated sharing of genes and phenotypic traits previously considered specific for
S. pneumoniae
hamper the exact identification of
S. pneumoniae
. Based on sequence analysis of 16S rRNA genes of a collection of 634 streptococcal strains, identified by multilocus sequence analysis, we detected a cytosine at position 203 present in all 440 strains of
S. pneumoniae
but replaced by an adenosine residue in all strains representing other species of mitis group streptococci. The
S. pneumoniae
-specific sequence signature could be demonstrated by sequence analysis or indirectly by restriction endonuclease digestion of a PCR amplicon covering the site. The
S. pneumoniae
-specific signature offers an inexpensive means for validation of the identity of clinical isolates and should be used as an integrated marker in the annotation procedure employed in 16S rRNA-based molecular studies of complex human microbiotas. This may avoid frequent misidentifications such as those we demonstrate to have occurred in previous reports and in reference sequence databases.
Cepheid Xpert MRSA Cycle Threshold in Discordant Colonization Results and as a Quantitative Measure of Nasal Colonization Burden
Authors: Stenehjem Edward , Rimland David , Crispell Emily K. , Stafford Cortney , Gaynes Robert , Satola Sarah W.
Published: March 2012

We analyzed the cycle threshold (

C
T

) of PCR surveillance MRSA swabs obtained from veterans. Lower

C
T

on admission was associated with a positive culture from nasal swabs at discharge. Compared to PCR, direct plating of nasal swabs performed poorly, especially for patients with an elevated

C
T

. The

C
T

is strongly correlated with quantitative nasal cultures. Clinical and infection control applications of the

C
T

have yet to be defined and warrant further evaluation.
Extensive Genetic Diversity within the Dutch Clinical Cryptococcus neoformans Population
Authors: Hagen Ferry , Illnait-Zaragozí María-Teresa , Meis Jacques F. , Chew William H. M. , Curfs-Breuker Ilse , Mouton Johan W. , Hoepelman Andy I. M. , Spanjaard Lodewijk , Verweij Paul E. , Kampinga Greetje A. , Kuijper Ed J. , Boekhout Teun , Klaassen Corné H. W.
Published: March 2012

A set of 300 Dutch
Cryptococcus neoformans
isolates, obtained from 237 patients during 1977 to 2007, was investigated by determining the mating type, serotype, and AFLP and microsatellite genotype and susceptibility to seven antifungal compounds. Almost half of the studied cases were from HIV-infected patients, followed by a patient group of individuals with other underlying diseases and immunocompetent individuals. The majority of the isolates were mating type α and serotype A, followed by αD isolates and other minor categories. The most frequently observed genotype was AFLP1, distantly followed by AFLP2 and AFLP3. Microsatellite typing revealed a high genetic diversity among serotype A isolates but a lower diversity within the serotype D set of isolates. One patient was infected by multiple AFLP genotypes. Fluconazole and flucytosine had the highest geometric mean MICs of 2.9 and 3.5 μg/ml, respectively, while amphotericin B (0.24 μg/ml), itraconazole (0.08 μg/ml), voriconazole (0.07 μg/ml), posaconazole (0.06 μg/ml), and isavuconazole (0.03 μg/ml) had much lower geometric mean MICs. One isolate had a high flucytosine MIC (>64 μg/ml), while decreased susceptibility (≥16 μg/ml) for flucytosine and fluconazole was found in 9 and 10
C. neoformans
isolates, respectively.
A New and Highly Divergent Enterocytozoon bieneusi Genotype Isolated from a Renal Transplant Recipient
Authors: Pomares Christelle , Santín Monica , Miegeville Michel , Espern Anne , Albano Laetitia , Marty Pierre , Morio Florent
Published: March 2012

A 49-year-old renal transplant recipient was admitted to our hospital due to abundant liquid diarrhea and dehydration. Parasitological investigations, including genotyping, led to the diagnosis of intestinal microsporidiosis due to a new and highly divergent internal transcribed spacer (ITS) genotype of
Enterocytozoon bieneusi
. The potential route of transmission through horse stools is discussed.
Human SCARB2-Dependent Infection by Coxsackievirus A7, A14, and A16 and Enterovirus 71
Authors: Yamayoshi Seiya , Iizuka Setsuko , Yamashita Teruo , Minagawa Hiroko , Mizuta Katsumi , Okamoto Michiko , Nishimura Hidekazu , Sanjoh Kanako , Katsushima Noriko , Itagaki Tsutomu , Nagai Yukio , Fujii Ken , Koike Satoshi
Published: March 2012

Human enterovirus species A (HEV-A) consists of at least 16 members of different serotypes that are known to be the causative agents of hand, foot, and mouth disease (HFMD), herpangina, and other diseases, such as respiratory disease and polio-like flaccid paralysis. Enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) are the major causative agents of HFMD. CVA5, CVA6, CVA10, and CVA12 mainly cause herpangina or are occasionally involved with sporadic cases of HFMD. We have previously shown that human scavenger receptor class B, member 2 (SCARB2) is a cellular receptor for EV71 and CVA16. Using a large number of clinical isolates of HEV-A, we explored whether all clinical isolates of EV71 and other serotypes of HEV-A infected cells via SCARB2. We tested this possibility by infecting L-SCARB2 cells, which are L929 cells expressing human SCARB2, by infecting human RD cells that had been treated with small interfering RNAs for SCARB2 and by directly binding the viruses to a soluble SCARB2 protein. We showed that all 162 clinical isolates of EV71 propagated in L-SCARB2 cells, suggesting that SCARB2 is the critical receptor common to all EV71 strains. In addition, CVA7, CVA14, and CVA16, which are most closely related to each other, also utilized SCARB2 for infection. EV71, CVA14, and CVA16 are highly associated with HFMD, and EV71 and CVA7 are occasionally associated with neurological diseases, suggesting that SCARB2 plays important roles in the development of these diseases. In contrast, another group of viruses, such as CVA2, CVA3, CVA4, CVA5, CVA6, CVA8, CVA10, and CVA12, which are relatively distant from the EV71 group, is associated mainly with herpangina. None of these clinical isolates infected via the SCARB2-dependent pathway. HEV-A viruses can be divided into at least two groups depending on the use of SCARB2, and the receptor usage plays an important role in developing the specific diseases for each group.
Kaposi's Sarcoma-Associated Herpesvirus Latency-Associated Nuclear Antigen Interacts with Multifunctional Angiogenin To Utilize Its Antiapoptotic Functions
Authors: Paudel Nitika , Sadagopan Sathish , Chakraborty Sayan , Sarek Grzegorz , Ojala Päivi M. , Chandran Bala
Published: March 2012

Kaposi's sarcoma-associated herpesvirus (KSHV) is etiologically associated with the angioproliferative Kaposi's sarcoma (KS). KSHV infection and the expression of latency-associated nuclear antigen (LANA-1) upregulates the angiogenic multifunctional 123-amino-acid, 14-kDa protein angiogenin (ANG), which is detected in KS lesions and in KSHV-associated primary effusion lymphoma (PEL) cells. ANG knockdown or the inhibition of ANG's nuclear translocation resulted in decreased LANA-1 gene expression and reduced KSHV-infected endothelial and PEL cell survival (Sadagopan et al., J. Virol.
83:
3342–3364, 2009). Further studies here demonstrate that LANA-1 and ANG colocalize and coimmunoprecipitate in
de novo
infected endothelial cells and in latently infected PEL (BCBL-1 and BC-3) cells. LANA-1 and ANG interaction occurred in the absence of the KSHV genome and other viral proteins. In gel filtration chromatography analyses of BC-3 cell lysates, ANG coeluted with LANA-1, p53, and Mdm2 in high-molecular-weight fractions, and LANA-1, p53, and Mdm2 also coimmunoprecipitated with ANG. LANA-1, ANG, and p53 colocalized in KSHV-infected cells, and colocalization between ANG and p53 was also observed in LANA-1-negative cells. The deletion constructs of ANG suggested that the C-terminal region of amino acids 104 to 123 is involved in LANA-1 and p53 interactions. Silencing ANG or inhibiting its nuclear translocation resulted in decreased nuclear LANA-1 and ANG levels, decreased interactions between ANG-LANA-1, ANG-p53, and LANA-1-p53, the induction of p53, p21, and Bax proteins, the increased cytoplasmic localization of p53, the downregulation of Bcl-2, the increased cleavage of caspase-3, and the apoptosis of cells. No such effects were observed in KSHV-negative BJAB cells. The phosphorylation of p53 at serine 15, which is essential for p53 stabilization and for p53's apoptotic and cell cycle regulation functions, was increased in BCBL-1 cells transduced with short hairpin RNA targeting ANG. Together, these studies suggest that the antiapoptosis observed in KSHV-infected cells and the suppression of p53 functions are mediated in part by ANG, and KSHV has probably evolved to utilize angiogenin's multiple functions for the maintenance of its latency and cell survival. Thus, targeting ANG to induce the apoptosis of cells latently infected with KSHV is an attractive therapeutic strategy against KSHV infection and associated malignancies.
Effects of Manipulating Apoptosis on Sindbis Virus Infection of Aedes aegypti Mosquitoes
Authors: Wang Hua , Gort Taryn , Boyle Daniel L. , Clem Rollie J.
Published: March 2012

Improved control of vector-borne diseases requires an understanding of the molecular factors that determine vector competence. Apoptosis has been shown to play a role in defense against viruses in insects and mammals. Although some observations suggest a correlation between apoptosis and resistance to arboviruses in mosquitoes, there is no direct evidence tying apoptosis to arbovirus vector competence. To determine whether apoptosis can influence arbovirus replication in mosquitoes, we manipulated apoptosis in
Aedes aegypti
mosquitoes by silencing the expression of genes that either positively or negatively regulate apoptosis. Silencing of the
A. aegypti
anti-apoptotic gene
iap1
(
Aeiap1
) caused apoptosis in midgut epithelium, alterations in midgut morphology, and 60 to 70% mosquito mortality. Mortality induced by
Aeiap1
silencing was rescued by cosilencing the initiator caspase gene
Aedronc
, indicating that the mortality was due to apoptosis. When mosquitoes which had been injected with
Aeiap1
double-stranded RNA (dsRNA) were orally infected with Sindbis virus (SINV), increased midgut infection and virus dissemination to other organs were observed. This increase in virus infection may have been due to the effects of widespread apoptosis on infection barriers or innate immunity. In contrast, silencing the expression of
Aedronc
, which would be expected to inhibit apoptosis, reduced SINV midgut infection and virus dissemination. Thus, our data suggest that some level of caspase activity and/or apoptosis may be necessary for efficient virus replication and dissemination in mosquitoes. This is the first study to directly test the roles of apoptosis and caspases in determining mosquito vector competence for arboviruses.