bluuebunny/crossref_metadata_2025 huggingface dataset:
The Production of Inequality: The Gender Division of Labor Across the Transition to Parenthood
Authors: Yavorsky Jill E. , Kamp Dush Claire M. , Schoppe‐Sullivan Sarah J.
Published: April 2015

Using longitudinal time diary and survey data from a community sample of dual‐earner couples across the transition to parenthood, the authors examined change in divisions of paid and unpaid work and assessed the accuracy of survey data for time use measurement. Mothers, according to the time diaries, shouldered the majority of child care and did not decrease their paid work hours. Furthermore, the gender gap was not present prebirth but emerged postbirth with women doing more than 2 hours of additional work per day compared to an additional 40 minutes for men. Moreover, the birth of a child magnified parents' overestimations of work in the survey data, and had the authors relied only on survey data, gender work inequalities would not have been apparent. The findings have important implications for (a) the state of the gender revolution among couples well positioned to obtain balanced workloads and (b) the utility of survey data to measure parents' division of labor.
The Nanochannel Delivery System for Constant Testosterone Replacement Therapy
Authors: Ferrati Silvia , Nicolov Eugenia , Zabre Erika , Geninatti Thomas , Shirkey Beverly A. , Hudson Lee , Hosali Sharath , Crawley Michael , Khera Mohit , Palapattu Ganesh , Grattoni Alessandro
Published: April 2015

Introduction

The goal of testosterone replacement is to provide long-term physiological supplementation at sufficient levels to mitigate the symptoms of hypogonadism.

Aim

The objective of this work is to determine if the implantable nanochannel delivery system (nDS) can present an alternative delivery strategy for the long-term sustained and constant release of testosterone.

Methods

A formulation of common testosterone esters (F1) was developed to enable nanochannel delivery of the low water soluble hormone. In vivo evaluation of testosterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels by liquid chromatography/mass spectrometry and a multiplex assay, respectively, in castrated Sprague-Dawley rats implanted with nDS-F1 implants or polymeric pellets was performed over a 6-month period. The percent of testosterone concentrations observed that fell within the normal range of testosterone levels for each animal was calculated and used to compare the study groups.

Main Outcome Measures

Sustain release of testosterone in vivo for over 6 months.

Results

The subcutaneous release of F1 from nDS implants exhibited sustained in vivo release kinetics and attained stable clinically relevant plasma testosterone levels. Plasma LH and FSH levels were significantly diminished in nDS-F1 implant–treated animals, confirming biological activity of the released testosterone.

Conclusions

In conclusion, we demonstrate that nDS-F1 implants represents a novel approach for the treatment of male hypogonadism. Further studies will be performed in view of translating the technology to clinical use.
The Immunotherapy of Canine Osteosarcoma: A Historical and Systematic Review
Authors: Wycislo K.L. , Fan T.M.
Published: April 2015

Osteosarcoma is a malignant mesenchymal neoplasm that accounts for the majority of primary bone tumors in dogs and shares biological and clinical similarities with osteosarcoma in humans. Despite dose intensification with conventional cytotoxic therapies, survival times for dogs and humans diagnosed with high‐grade osteosarcoma have not changed in the past 20 years, with the principal cause of mortality being the development of pulmonary metastases. Given the therapeutic plateau reached for delaying metastatic progression with cytotoxic agents, exploration of alterative adjuvant therapies for improving management of osteosarcoma micrometastases is clinically justified. Evidence suggests that osteosarcoma is an immunogenic tumor, and development of immunotherapies for the treatment of microscopic lung metastases might improve long‐term outcomes. In this review, the history and foundational knowledge of immune interactions to canine osteosarcoma are highlighted. In parallel, immunotherapeutic strategies that have been explored for the treatment of canine osteosarcoma are summarized. With a greater understanding and awareness for how the immune system might be redirected toward combating osteosarcoma metastases, the rational development of diverse immune strategies for managing osteosarcoma holds substantial promise for transforming the therapeutic landscape and improving disease management in both dogs and human beings.
The Messy Practice of Building Women's Human Rights: Truth‐telling and Sexual Violence inGuatemala
Authors: Rosser Emily
Published: April 2015

This article seeks to contribute to broader discussions of how human rights emerge and become intelligible in moments of transition at national and transnational levels. It focuses on ad hoc practices of exposing war‐time sexual violence in two major truth‐telling processes inGuatemala, theRecuperation ofHistoricalMemoryProject and theCommission forHistoricalClarification. These initiatives collected testimony to document thousands of cases of state‐sanctioned violence during the 36 years of war that officially concluded with the peace accords inDecember 1996. Among their other achievements, these processes were the first truth‐telling bodies inLatinAmerica to expose and condemn gender violence in war. InGuatemala, the postwar transition occurred at precisely the moment that the organizing of women internationally was beginning to bear fruit at the statutory level, but the feminist analysis that developed is not attributable merely to the uptake of developments in international law. This article considers theseGuatemalan cases to argue that, although legal progress is crucial, practice has been a key alternative source for the development of women's human rights around sexual violence.
Writing Language‐based Trade Books: Making Linguistics Accessible to Lay Audiences
Authors: Reaser Jeffrey , Myrick Caroline
Published: April 2015

New publishing models and increased interest in “popular science” books have converged to make it increasingly viable for linguists to write language‐based trade books focusing on regional and social speech varieties. In order for such books to reach a broad audience, authors must make writing choices that are quite dissimilar from those made when writing for academic peers. In this article, we examine a number of recent sociolinguistic trade books and consider the effects of the authors' choices on the effectiveness in engaging and conveying technical information to non‐academic audiences. From our investigation of issues of audience, prose, images, citations, etc., emerge some guidelines that might help others be more successful in appealing to a popular readership. Though we offer no prescriptions, we hope that our investigation begins a conversation of best practices for engaging the public in linguistic investigations of regional and social speech varieties via trade books. In doing so, we also examine issues related to research ethics and the value in the academy of trade books specifically and linguistic gratuity projects in general.
EFL Learners' Multiple Documents Literacy: Effects of a Strategy‐Directed Intervention Program
Authors: Karimi Mohammad Nabi
Published: April 2015

There is a substantial body of L2 research documenting the central role of strategy instruction in reading comprehension. However, this line of research has been conducted mostly within the single text paradigm of reading research. With reading literacy undergoing a marked shift from single source reading to multiple documents literacy, little is known about the role of strategy instruction in readers' performance in this emerging type of literacy and the likelihood of readers' effective transfer of strategies from the single text‐dominated reading in instructional programs to the multiple texts‐dominated reading within academic knowledge societies. This study was conducted to begin to address this gap. 76 EFL learners were randomly assigned to one of four conditions: ST/−SI (single text/no strategy instruction), ST/+SI (single text/strategy instruction), MT/−SI (multiple texts/no strategy instruction) and MT/+SI (multiple texts/strategy instruction). Pretests of reading comprehension and self‐reported reading strategy use confirmed that the four groups performed at similar levels. They were then taught for 10–90‐minute sessions and were tested for multiple texts comprehension and strategy use. The results revealed the significant effects of strategy instruction on the participants' multiple documents comprehension and strategy use and the more positive effect of task‐relevant strategy instruction on multiple documents comprehension.
No increase in prevalence of somatization in functional vs organic dyspepsia: a cross‐sectional survey
Authors: Gracie D. J. , Bercik P. , Morgan D. G. , Bolino C. , Pintos‐Sanchez M. I. , Moayyedi P. , Ford A. C.
Published: April 2015

Background

Psychological factors are associated with functional gastrointestinal (GI) disorders. Literature suggests that somatization is associated with functional dyspepsia (FD). However, the relationship between organic dyspepsia (OD), FD, and FD subtypes and somatization is poorly described. We aimed to examine this issue in a cross‐sectional study of secondary care patients.

Methods

Demographic and GI symptom data were collected from 4224 adult patients via the Rome III questionnaire. Somatization data were collected using the patient health questionnaire‐12. Mean somatization score and number of somatic symptoms were compared between patients with organic and FD, and between FD subtypes using analysis of variance. The same comparison was undertaken for the proportion of patients reporting individual somatic symptoms.

Key Results

Exactly, 783 patients met criteria for dyspepsia, of whom 231 (29.5%) had organic disease following upper GI endoscopy. Mean somatization scores and number of somatic symptoms were no higher in functional vs OD (p = 0.23; p = 0.19). In addition, while the prevalence of somatization in FD was relatively high, there was no difference in severity of somatization in FD subgroups.

Conclusions & Inferences

Somatization is associated with functional and OD to the same degree. Overall severity of somatization did not appear to vary according to FD subtype.
Intraoperative changes in blood pressure associated with cerebral desaturation in infants
Authors: Michelet Daphné , Arslan Ozkan , Hilly Julie , Mangalsuren Nyamjargal , Brasher Christopher , Grace Robert , Bonnard Arnaud , Malbezin Serge , Nivoche Yves , Dahmani Souhayl
Published: April 2015

Background

Intraoperative hypotension has been linked to poor postoperative neurological outcomes. However, the definition of hypotension remains controversial in children. We sought to determine arterial blood pressure threshold values associated with cerebral desaturation in infants.

Methods

After ethics committee approval, infants younger than 3 months were included in this prospective observational study. Cerebral saturation was assessed using near‐infrared spectroscopy. The primary goal of the study was to determine percentage reductions in intraoperative systolic blood pressure (SBP) and mean blood pressure (MBP) associated with decreases in cerebral blood oxygen saturation of >20%, when compared to baseline. Analyses were performed using a bootstrap receiving operator characteristic (ROC) curves with determination of the gray zone.

Results

Sixty patients were recruited and 960 measurement points were recorded. Fifty‐nine data points (6.1%) recorded cerebral desaturation of >20% when compared to baseline. The areas under the ROC curves were 0.79 (0.74–0.84) and 0.67 (0.6–0.75) for percentage decreases in SBP and MBP, respectively. Gray zone values with false‐positive and negative rates <10% were SBP decreases of 20.5% and 37.5%, respectively, and MBP decreases of 15.5% and 44.5%, respectively.

Conclusion

Our results indicate that falls in noninvasive systolic blood pressure of <20% from baseline are associated with a <10% chance of cerebral desaturation in neonates and infants <3 months of age undergoing noncardiac surgery. As such, maintaining systolic blood pressure above this threshold value appears a valid clinical target.
Survey research: it's just a few questions, right?
Authors: Tait Alan R. , Voepel‐Lewis Terri
Published: April 2015

While most anesthesiologists and other physician‐ or nurse‐scientists are familiar with traditional descriptive, observational, and interventional study design, survey research has typically remained the preserve of the social scientists. To that end, this article provides a basic overview of the elements of good survey design and offers some rules of thumb to help guide investigators through the survey process.
Improved method for measuring the apparent CO2 photocompensation point resolves the impact of multiple internal conductances to CO2 to net gas exchange
Authors: Walker Berkley J. , Ort Donald R.
Published: April 2015

There is a growing interest in accurate and comparable measurements of the CO2 photocompensation point (Γ*), a vital parameter to model leaf photosynthesis. The Γ* is measured as the common intersection of several CO2 response curves, but this method may incorrectly estimate Γ* by using linear fits to extrapolate curvilinear responses and single conductances to convert intercellular photocompensation points (Ci*) to chloroplastic Γ*. To determine the magnitude and minimize the impact of these artefacts on Γ* determinations, we used a combination of meta‐analysis, modelling and original measurements to develop a framework to accurately determine Ci*. Our modelling indicated that the impact of using linear fits could be minimized based on the measurement CO2 range. We also propose a novel method of analysing common intersection measurements using slope–intercept regression. Our modelling indicated that slope–intercept regression is a robust analytical tool that can help determine if a measurement is biased because of multiple internal conductances to CO2. Application of slope–intercept regression to Nicotiana tabacum and Glycine max revealed that multiple conductances likely have little impact to Ci* measurements in these species. These findings present a robust and easy to apply protocol to help resolve key questions concerning CO2 conductance through leaves.
Toward an Anthropology of War Propaganda
Authors: Kiper Jordan
Published: April 2015

Prior to the Rwandan genocide, the study of war propaganda had all but disappeared as a significant topic of interest for lawyers and social scientists. However, since the trials of war propagandists by international criminal tribunals, the study has been reignited. The reason is due to the manner in which legal actors discussed the effects of war propaganda and pronounced its criminality. They claimed that war propaganda constitutes incitement not only because it attempts to foment dangerously violent ideologies, but also because it actually causes mass violence. In defining war propaganda in this way, tribunals have shifted the crime of incitement from being inchoate to causal. This new precedent has led ethnographers to investigate the manner in which war propaganda has related to mass violence and to challenge the tribunal's purported causal link. Additionally, it has led legal researchers to generate novel theories about war propaganda that are conducive to the new precedent but would still benefit from the frameworks and methodologies of anthropology.
Experimental and Numerical Investigations Using Lattice Boltzmann Method to Study Shedding Vortices in an Unsteady Confined Flow Around an Obstacle
Authors: Wafik Abassi , Fethi Aloui , Keirsbulck Laurent , Sassi Ben Nasrallah
Published: April 2015

Lattice Boltzmann equation with Bhatnagar–Gross–Krook (BGK) model is applied to simulate unsteady laminar flow around a confined square obstacle, in order to study the vortex shedding and their interaction in the flow on the mass transfer in the parietal zone of a channel. The model was tested by comparing to an experimental study via standard particle image velocimetry (PIV). A post-processing was used to well extract instantaneous vortices contained in the flow downstream obstacles. A sensor with zero concentration on the surface is placed on the channel wall to study the effect of wake instabilities on the parietal mass transfer.
Poiseuille Flow and Thermal Transpiration of a Rarefied Gas Between Parallel Plates: Effect of Nonuniform Surface Properties of the Plates in the Transverse Direction
Authors: Doi Toshiyuki
Published: April 2015

Poiseuille flow and thermal transpiration of a rarefied gas between parallel plates with nonuniform surface properties in the transverse direction are studied based on kinetic theory. We considered a simplified model in which one wall is a diffuse reflection boundary and the other wall is a Maxwell-type boundary on which the accommodation coefficient varies periodically and smoothly in the transverse direction. The spatially two-dimensional (2D) problem in the cross section is studied numerically based on the linearized Bhatnagar–Gross–Krook–Welander (BGKW) model of the Boltzmann equation. The flow behavior, i.e., the macroscopic flow velocity and the mass flow rate of the gas as well as the velocity distribution function, is studied over a wide range of the mean free path of the gas and the parameters of the distribution of the accommodation coefficient. The mass flow rate of the gas is approximated by a simple formula consisting of the data of the spatially one-dimensional (1D) problems. When the mean free path is large, the distribution function assumes a wavy variation in the molecular velocity space due to the effect of a nonuniform surface property of the plate.
Effects of Rotating Inlet Distortion on Compressor Stability With Stall Precursor-Suppressed Casing Treatment
Authors: Dong Xu , Sun Dakun , Li Fanyu , Jin Donghai , Gui Xingmin , Sun Xiaofeng
Published: April 2015

This paper conducts an experimental research of rotating inlet distortion on a low-speed large size test compressor with emphasis on the stability problem of axial fan/compressors, and the stall margin enhancement with a kind of stall precursor-suppressed (SPS) casing treatment. Some results on compressor stall margin and prestall behavior under the restriction of rotating inlet distortion are presented. The experimental results show that whether the inlet distortion is co-rotating or counter-rotating, the SPS casing treatment can still improve the stall margin without leading to additional efficiency loss caused by such configuration. The experiment results also show that the mechanism of the stall margin improvement with such casing treatment is associated with delaying the nonlinear development of the stall precursor waves and weakening the unsteady flow disturbances in a compression system.
The Single-Blow Problem Including the Effects of Longitudinal Conduction
Authors: Howard C. P.
Published: April 2015

The results are presented from a numerical finite-difference method of calculation for the transient behavior of porous media when subjected to a step change in fluid temperature considering the case where the longitudinal thermal heat conduction cannot be neglected. These results, given in tabular and graphical form, provide a useful means for evaluating the heat-transfer data obtained from the transient testing of compact heat-exchanger surfaces.
The Lift/Cruise Fan Propulsion System for VTOL Applications
Authors: Kutney John T.
Published: April 2015

The high by-pass ratio tip-turbine driven lift and lift cruise fan offers the unique arrangement of a single type propulsion system fulfilling the many requirements for a VTOL application. This simple, compact, lightweight system offers high efficiency (low SFC) with the high static thrust at take-off required for VTOL and STOL operation. The lift cruise fan is a logical outgrowth of the tip-turbine lift fan technology backed by over 600 hr of testing since 1959. The significant test problem accomplishments for the cruise fan are presented as well as a representative VTOL transport-type application.
Controls Integrate the Modular On-Site Gas-Turbine Energy System
Authors: Hinde William L. , Williams Arthur H.
Published: April 2015

Of growing significance is the conversion of natural gas into electricity, on-site, through the use of direct-fired gas turbines. Most installations will best be served by applying the “modular concept”; i.e., a group of turbine-alternator modules which can be added or taken from the line to meet the variation in load profile throughout a 24-hr day. Many of these must operate on a completely automatic, unattended basis. The purpose of the paper is to outline the various automatic controls required, describe some of the major factors in the design of these controls, and to provide a better understanding of the completely automatic, modular on-site gas-turbine energy system.
Installation and Operation of the Model LM100 Turboshaft Engine Aboard the HS “Denison”
Authors: Kerr W. A.
Published: April 2015

This paper reviews the modification, installation and utilization of an aircraft turboshaft engine in a hydrofoil ship. A detailed description of engine performance, maintenance, and corrosion protection is included.
Evaluation of Air Filter Test Methods
Authors: May James W.
Published: April 2015

There are three generally accepted test methods in current use for determining the effectiveness of air filters used for the collection of atmospheric dust. These three methods are (a) weight, (b) dust spot, and (c) DOP, which uses dioctyl-phylate as the aerosol. These test methods are discussed briefly and illustrations are used to assist the reader in visualizing the concentration of dust which will remain in the clean-air stream for efficiency values reported by each test method.
Use of Scale-Model Aids in Solution of Complex Air-Flow Design Problem
Authors: Oppenheim Arthur , Oran Frederic M.
Published: April 2015

This paper describes a method used to design the inlet section of a jet-engine test cell. Calculations, accomplished with the use of standard, tabulated flow formulas and coefficients which can be found in basic fluid-flow texts, are shown. The step-by-step procedure enables the reader to use the methods and formulas in this paper as a guide for solving similar problems. Selection of an optimum turning-vane configuration is described herein by the use of tufts of cotton in the air stream of the model. A table is included which compares the model data with actual full-scale construction at various stages.
Heat Recovery Problems in Total Energy Systems
Authors: Boyen John L.
Published: April 2015

This paper is devoted to a discussion of problems and factors incidental to the design and selection of exhaust heat-recovery equipment applicable to total energy systems, based on the gas turbine as the prime mover and source of recoverable heat energy. Factors such as type of equipment, form of heat exchanger, ideal performance, and typical design considerations are included. This also covers several forms of the basic exhaust heat-recovery unit, including the addition of supplementary firing, fired and unfired superheaters, and cycles for power-conversion applications.
Marinization of the General Electric LM 1500 Gas Turbine
Authors: Stoeckly E. E.
Published: April 2015

After presenting some of the attractive features and characteristics of the aircraft gas turbine for selected marine propulsion applications, this paper looks at those features of such engines that need improvement in order that they may become true marine power plants. Both cold and hot-corrosion problems are created by the marine atmosphere and the change from aircraft-type fuels to higher-sulfur-content marine fuels. Phases of a development program to produce the required features in the General Electric LM 1500 gas-turbine power plant are discussed.
Analysis and Testing of Air-Cooled Turbine Rotor and Stator Blades
Authors: Helms H. E. , Emmerson C. W.
Published: April 2015

Advancing turbine engine technology requires air-cooled turbines. Cooling mechanisms applied must be exploited in a practical manner to obtain maximum cooling effectiveness. Cooled turbine stator and rotor blade design requires rigorous analysis supplemented by verifying experimental data. Problem definition, analysis techniques, material application, cascade and engine testing, and correlation of data are presented for air-cooled turbines. Convection, impingement, film, transpiration, and combined cooling mechanisms are reviewed.
Discrete Frequency Noise Generation due to Fluid Flow Over Blades, Supporting Spokes, and Similar Bodies
Authors: Parker R.
Published: April 2015

The author reviews, briefly, previous work on the excitation of powerful acoustic resonances by periodicity in the wakes shed by blades, flat plates, spokes, or other similar bodies in a fluid flow. He presents further experimental data, from low-speed wind-tunnel tests, that add to the understanding of the phenomenon, but there is still a great deal that is not understood regarding the mechanism of excitation. While mechanical vibration can be excited, it has been established conclusively that this does not play any part in generating the intense discrete frequency noise observed in many cases. In the present tests, the amplitude of forced vibration of a plate due to the pressure fluctuations at an acoustic resonance was an order of magnitude greater than the amplitude of the directly excited mechanical resonance.
The Potential of the Gas-Turbine Vehicle in Alleviating Air Pollution
Authors: Wright Edward S.
Published: April 2015

The author examines the potential of the gas turbine in alleviating air pollution as a replacement for presently used vehicular reciprocating engines. Automobiles receive particular emphasis because of the magnitude of their contribution to the problem. Emissions of gas-turbine engines are compared with those of gasoline reciprocating engines, and other important characteristics relevant to the problem (such as engine power output, reliability, transmissions, vehicle performance, fuel consumption, and manufacturing cost) are discussed. It is concluded that widespread adoption of vehicular gas-turbine engines can — provided these engines can be produced and operated at costs competitive with those of future reciprocating engines — can virtually eliminate automotive air pollution as a source of serious environmental concern.
Proposed Gas Turbine Procurement Standard: Gaseous and Liquid Fuels
Authors: Holzwarth H. T. , Howard G. S. , Stott T. E.
Published: April 2015

Since fuel comprises a fairly large percent of total gas-turbine operating costs, it is an extremely important subject to the user. This paper, one of a series (see Appendix), describes allowable specifications for gaseous and liquid fuels suitable for burning in gas turbines.
Rehabilitation Following Proximal Rectus Femoris Repair: A Case Report
Authors: Adler Kelly L. , Cook P. Christopher , Giordano Brian D.
Published: April 2015

Injury to the rectus femoris (RF) myotendinous complex is the most common location of quadriceps injury, due to combined loads of stretch and eccentric muscular activation. To our knowledge, open proximal RF repair has been reported, but a thorough description of postoperative rehabilitation and functional progression of athletic activity has not been described. This case report outlines the rehabilitation of a 30-year-old female following open proximal RF repair after 15 months of failed conservative treatment. Six months postoperatively she returned to competitive recreational soccer with no complaints.
Nepal disaster presages a coming megaquake
Authors: Hand Eric , Pulla Priyanka
Published: April 2015

Rupture in quiet region of crust suggests Himalayan threat was underestimated.
Extinction risks from climate change
Authors: Lambers Janneke Hille Ris
Published: April 2015

How will climate change affect global biodiversity?

[Also see Report by

Urban
et al.

]
Seeing optics in a new light
From Sight to Light The Passage from Ancient to Modern Optics
A. Mark Smith
University of Chicago Press, 2015. 469 pp.
Authors: Dainty Christopher
Published: April 2015

Johannes Kepler and the transition to modern optical theories
Dysregulation of Serum Gamma Interferon Levels in Vascular Chronic Q Fever Patients Provides Insights into Disease Pathogenesis
Authors: Pennings Jeroen L. A. , Kremers Marjolein N. T. , Hodemaekers Hennie M. , Hagenaars Julia C. J. P. , Koning Olivier H. J. , Renders Nicole H. M. , Hermans Mirjam H. A. , de Klerk Arja , Notermans Daan W. , Wever Peter C. , Janssen Riny
Published: April 2015

A large community outbreak of Q fever occurred in the Netherlands in the period 2007 to 2010. Some of the infected patients developed chronic Q fever, which typically includes pathogen dissemination to predisposed cardiovascular sites, with potentially fatal consequences. To identify the immune mechanisms responsible for ineffective clearance ofCoxiella burnetiiin patients who developed chronic Q fever, we compared serum concentrations of 47 inflammation-associated markers among patients with acute Q fever, vascular chronic Q fever, and past resolved Q fever. Serum levels of gamma interferon were strongly increased in acute but not in vascular chronic Q fever patients, compared to past resolved Q fever patients. Interleukin-18 levels showed a comparable increase in acute as well as vascular chronic Q fever patients. Additionally, vascular chronic Q fever patients had lower serum levels of gamma interferon-inducible protein 10 (IP-10) and transforming growth factor β (TGF-β) than did acute Q fever patients. Serum responses for these and other markers indicate that type I immune responses toC. burnetiiare affected in chronic Q fever patients. This may be attributed to an affected immune system in cardiovascular patients, which enables localC. burnetiireplication at affected cardiovascular sites.
Development and Evaluation of a Multiplex Microsphere Assay for Quantitation of IgG and IgA Antibodies against Neisseria meningitidis Serogroup A, C, W, and Y Polysaccharides
Authors: Bårnes Guro K. , Kristiansen Paul A. , Caugant Dominique A. , Næss Lisbeth M.
Published: April 2015

We developed and evaluated a rapid and simple multiplex microsphere assay for the quantification of specific IgG and IgA antibodies against meningococcal serogroup A, C, W, and Y capsular polysaccharides in serum and saliva. Meningococcal polysaccharides were conjugated to distinct magnetic carboxylated microspheres, and the performance of the assay was assessed using the CDC1992 standard meningococcal reference serum and a panel of serum and saliva samples. The standard curve was linear over an eight 3-fold dilution range in the IgG assay and a seven 3-fold dilution range in the IgA assay. No cross-reactivity was discovered, and the assay showed high specificity with ≥91% homologous inhibition and ≤11% heterologous inhibition for all serogroups and immunoglobulin classes. Lower limits of detections were ≤280 pg/ml for IgG and ≤920 pg/ml for IgA antibodies. The assay was reproducible, with a mean coefficient of variation of ≤5% for intra-assay duplicates, a mean coefficient of variation of ≤20% for interassay repeated analysis with different conjugations of microspheres, and a mean coefficient of variation within 25.8% for interoperator variation. The assay showed good correlation to the standard meningococcal polysaccharide enzyme-linked immunosorbent assay (ELISA) for detection of serum antibodies. This multiplex assay is robust and reliable and requires less sample volume, and less time and workload are needed than for ELISA, making this method highly relevant for serological and salivary investigations on the effect of meningococcal vaccines and for immunosurveillance studies.
Contamination of Ambient Air with Acinetobacter baumannii on Consecutive Inpatient Days
Authors: Shimose Luis A. , Doi Yohei , Bonomo Robert A. , De Pascale Dennise , Viau Roberto A. , Cleary Timothy , Namias Nicholas , Kett Daniel H. , Munoz-Price L. Silvia
Published: April 2015

Acinetobacter
-positive patients had their ambient air tested for up to 10 consecutive days. The air was
Acinetobacter
positive for an average of 21% of the days; the rate of contamination was higher among patients colonized in the rectum than in the airways (relative risk [RR], 2.35;
P
= 0.006). Of the 6 air/clinical isolate pairs available, 4 pairs were closely related according to rep-PCR results.
Efficient Detection of Carbapenemase Activity in Enterobacteriaceae by Matrix-Assisted Laser Desorption Ionization−Time of Flight Mass Spectrometry in Less Than 30 Minutes
Authors: Lasserre Camille , De Saint Martin Luc , Cuzon Gaelle , Bogaerts Pierre , Lamar Estelle , Glupczynski Youri , Naas Thierry , Tandé Didier
Published: April 2015

The recognition of carbapenemase-producing
Enterobacteriaceae
(CPE) isolates is a major laboratory challenge, and their inappropriate or delayed detection may have negative impacts on patient management and on the implementation of infection control measures. We describe here a matrix-assisted laser desorption ionization−time of flight (MALDI-TOF)-based method to detect carbapenemase activity in
Enterobacteriaceae
. After a 20-min incubation of the isolate with 0.5 mg/ml imipenem at 37°C, supernatants were analyzed by MALDI-TOF in order to identify peaks corresponding to imipenem (300 Da) and an imipenem metabolite (254 Da). A total of 223 strains, 77 CPE (OXA-48 variants, KPC, NDM, VIM, IMI, IMP, and NMC-A) and 146 non-CPE (cephalosporinases, extended-spectrum β-lactamases [ESBLs], and porin defects), were tested and used to calculate a ratio of imipenem hydrolysis: mass spectrometry [MS] ratio = metabolite/(imipenem + metabolite). An MS ratio cutoff was statistically determined to classify strains as carbapenemase producers (MS ratio of ≥0.82). We validated this method first by testing 30 of our 223 isolates (15 CPE and 15 non-CPE) 10 times to calculate an intraclass correlation coefficient (ICC of 0.98), showing the excellent repeatability of the method. Second, 43 strains (25 CPE and 18 non-CPE) different from the 223 strains used to calculate the ratio cutoff were used as external controls and blind tested. They yielded sensitivity and specificity of 100%. The total cost per test is <0.10 U.S. dollars (USD). This easy-to-perform assay is time-saving, cost-efficient, and highly reliable and might be used in any routine laboratory, given the availability of mass spectrometry, to detect CPE.
Naturally Occurring Resistance-Associated Variants of Hepatitis C Virus Protease Inhibitors in Poor Responders to Pegylated Interferon-Ribavirin
Authors: Larrat Sylvie , Vallet Sophie , David-Tchouda Sandra , Caporossi Alban , Margier Jennifer , Ramière Christophe , Scholtes Caroline , Haïm-Boukobza Stéphanie , Roque-Afonso Anne-Marie , Besse Bernard , André-Garnier Elisabeth , Mohamed Sofiane , Halfon Philippe , Pivert Adeline , LeGuillou-Guillemette Hélène , Abravanel Florence , Guivarch Matthieu , Mackiewicz Vincent , Lada Olivier , Mourez Thomas , Plantier Jean-Christophe , Baazia Yazid , Alain Sophie , Hantz Sebastien , Thibault Vincent , Gaudy-Graffin Catherine , Bouvet Dorine , Mirand Audrey , Henquell Cécile , Gozlan Joel , Lagathu Gisèle , Pronier Charlotte , Velay Aurélie , Schvoerer Evelyne , Trimoulet Pascale , Fleury Hervé , Bouvier-Alias Magali , Brochot Etienne , Duverlie Gilles , Maylin Sarah , Gouriou Stéphanie , Pawlotsky Jean-Michel , Morand Patrice
Published: April 2015

The pretherapeutic presence of protease inhibitor (PI) resistance-associated variants (RAVs) has not been shown to be predictive of triple-therapy outcomes in treatment-naive patients. However, they may influence the outcome in patients with less effective pegylated interferon (pegIFN)-ribavirin (RBV) backbones. Using hepatitis C virus (HCV) population sequence analysis, we retrospectively investigated the prevalence of baseline nonstructural 3 (NS3) RAVs in a multicenter cohort of poor IFN-RBV responders (i.e., prior null responders or patients with a viral load decrease of <1 log IU/ml during the pegIFN-RBV lead-in phase). The impact of the presence of these RAVs on the outcome of triple therapy was studied. Among 282 patients, the prevalances (95% confidence intervals) of baseline RAVs ranged from 5.7% (3.3% to 9.0%) to 22.0% (17.3% to 27.3%), depending to the algorithm used. Among mutations conferring a >3-fold shift in 50% inhibitory concentration (IC
50
) for telaprevir or boceprevir, T54S was the most frequently detected mutation (3.9%), followed by A156T, R155K (0.7%), V36M, and V55A (0.35%). Mutations were more frequently found in patients infected with genotype 1a (7.5 to 23.6%) than 1b (3.3 to 19.8%) (
P
= 0.03). No other sociodemographic or viroclinical characteristic was significantly associated with a higher prevalence of RAVs. No obvious effect of baseline RAVs on viral load was observed. In this cohort of poor responders to IFN-RBV, no link was found with a sustained virological response to triple therapy, regardless of the algorithm used for the detection of mutations. Based on a cross-study comparison, baseline RAVs are not more frequent in poor IFN-RBV responders than in treatment-naive patients and, even in these difficult-to-treat patients, this study demonstrates no impact on treatment outcome, arguing against resistance analysis prior to treatment.
Kallikrein-8 Proteolytically Processes Human Papillomaviruses in the Extracellular Space To Facilitate Entry into Host Cells
Authors: Cerqueira Carla , Samperio Ventayol Pilar , Vogeley Christian , Schelhaas Mario
Published: April 2015

The entry of human papillomaviruses into host cells is a complex process. It involves conformational changes at the cell surface, receptor switching, internalization by a novel endocytic mechanism, uncoating in endosomes, trafficking of a subviral complex to the Golgi complex, and nuclear entry during mitosis. Here, we addressed how the stabilizing contacts in the capsid of human papillomavirus 16 (HPV16) may be reversed to allow uncoating of the viral genome. Using biochemical and cell-biological analyses, we determined that the major capsid protein L1 underwent proteolytic cleavage during entry. In addition to a dispensable cathepsin-mediated proteolysis that occurred likely after removal of capsomers from the subviral complex in endosomes, at least two further proteolytic cleavages of L1 were observed, one of which was independent of the low-pH environment of endosomes. This cleavage occurred extracellularly. Further analysis showed that the responsible protease was the secreted trypsin-like serine protease kallikrein-8 (KLK8) involved in epidermal homeostasis and wound healing. Required for infection, the cleavage was facilitated by prior interaction of viral particles with heparan sulfate proteoglycans. KLK8-mediated cleavage was crucial for further conformational changes exposing an important epitope of the minor capsid protein L2. Occurring independently of cyclophilins and of furin that mediate L2 exposure, KLK8-mediated cleavage of L1 likely facilitated access to L2, located in the capsid lumen, and potentially uncoating. Since HPV6 and HPV18 also required KLK8 for entry, we propose that the KLK8-dependent entry step is conserved.

IMPORTANCEOur analysis of the proteolytic processing of incoming HPV16, an etiological agent of cervical cancer, demonstrated that the capsid is cleaved extracellularly by a serine protease active during wound healing and that this cleavage was crucial for infection. The cleavage of L1 is one of at least four structural alterations that prime the virus extracellularly for receptor switching, internalization, and possibly uncoating. This step was also important for HPV6 and HPV18, which may suggest that it is conserved among the papillomaviruses. This study advances the understanding of how HPV16 initially infects cells, strengthens the notion that wounding facilitates infection of epidermal tissue, and may help the development of antiviral measures.
Identification of a Novel Hepacivirus in Domestic Cattle from Germany
Authors: Baechlein Christine , Fischer Nicole , Grundhoff Adam , Alawi Malik , Indenbirken Daniela , Postel Alexander , Baron Anna Lena , Offinger Jennifer , Becker Kathrin , Beineke Andreas , Rehage Juergen , Becher Paul
Published: April 2015

Hepatitis C virus (HCV) continues to represent one of the most significant threats to human health. In recent years, HCV-related sequences have been found in bats, rodents, horses, and dogs, indicating a widespread distribution of hepaciviruses among animals. By applying unbiased high-throughput sequencing, a novel virus of the genus
Hepacivirus
was discovered in a bovine serum sample.
De novo
assembly yielded a nearly full-length genome coding for a polyprotein of 2,779 amino acids. Phylogenetic analysis confirmed that the virus represents a novel species within the genus
Hepacivirus
. Viral RNA screening determined that 1.6% (
n =
5) of 320 individual animals and 3.2% (
n =
5) of 158 investigated cattle herds in Germany were positive for bovine hepacivirus. Repeated reverse transcription-PCR (RT-PCR) analyses of animals from one dairy herd proved that a substantial percentage of cows were infected, with some of them being viremic for over 6 months. Clinical and postmortem examination revealed no signs of disease, including liver damage. Interestingly, quantitative RT-PCR from different organs and tissues, together with the presence of an miR-122 binding site in the viral genome, strongly suggests a liver tropism for bovine hepacivirus, making this novel virus a promising animal model for HCV infections in humans.

IMPORTANCE
Livestock animals act as important sources for emerging pathogens. In particular, their large herd size and the existence of multiple ways of direct and food-borne infection routes emphasize their role as virus reservoirs. Apart from the search for novel viruses, detailed characterization of these pathogens is indispensable in the context of risk analysis. Here, we describe the identification of a novel HCV-like virus in cattle. In addition, determination of the prevalence and of the course of infection in cattle herds provides valuable insights into the biology of this novel virus. The results presented here form a basis for future studies targeting viral pathogenesis of bovine hepaciviruses and their potential to establish zoonotic infections.
Human Endogenous Retrovirus Type K (HERV-K) Particles Package and Transmit HERV-K–Related Sequences
Authors: Contreras-Galindo Rafael , Kaplan Mark H. , Dube Derek , Gonzalez-Hernandez Marta J. , Chan Susana , Meng Fan , Dai Manhong , Omenn Gilbert S. , Gitlin Scott D. , Markovitz David M.
Published: April 2015

Human endogenous retroviruses (HERV) make up 8% of the human genome. While the youngest of these retroviruses, HERV-K(HML-2), termed HK2, is able to code for all viral proteins and produce virus-like particles, it is not known if these virus particles package and transmit HK2-related sequences. Here, we analyzed the capacity of HK2 for packaging and transmitting HK2 sequences. We created an HK2 probe, termed Bogota, which can be packaged into HK2 viruses, and transfected it into cells that make HK2 particles. Supernatants of the transfected cells, which contained HK2 viral particles, then were added to target cells, and the transmissibility of the HK2 Bogota reporter was tracked by G418 resistance. Our studies revealed that contemporary HK2 virions produced by some teratocarcinoma and breast cancer cell lines, as well as by peripheral blood lymphocytes from lymphoma patients, can package HK2 Bogota probes, and these viruses transmitted these probes to other cells. After transmission, HK2 Bogota transcripts undergo reverse transcription, a step impaired by antiretroviral agents or by introduction of mutations into the probe sequences required for reverse transcription. HK2 viruses were more efficiently transmitted in the presence of HK2 Rec or HIV-1 Tat and Vif. Transmitted Bogota probes formed episomes but did not integrate into the cellular genome. Resistance to integration might explain the relatively low number of HK2 insertions that were acquired during the last 25 million years of evolution. Whether transient transmission of modern HK2 sequences, which encode two putative oncoproteins, can lead to disease remains to be studied.

IMPORTANCERetroviruses invaded the genome of human ancestors over the course of millions of years, yet these viruses generally have been inactivated during evolution, with only remnants of these infectious sequences remaining in the human genome. One of these viruses, termed HK2, still is capable of producing virus particles, although these particles have been regarded as being noninfectious. Using a genetic probe derived from HK2, we have discovered that HK2 viruses produced in modern humans can package HK2 sequences and transmit them to various other cells. Furthermore, the genetic sequences packaged in HK2 undergo reverse transcription. The transmitted probe circularized in the cell and failed to integrate into the cellular genome. These findings suggest that modern HK2 viruses can package viral RNA and transmit it to other cells. Contrary to previous views, we provide evidence of an extracellular viral phase of modern HK2 viruses. We have no evidence of sustained, spreading infection.
A Functional Interaction between Herpes Simplex Virus 1 Glycoprotein gH/gL Domains I and II and gD Is Defined by Using Alphaherpesvirus gH and gL Chimeras
Authors: Fan Qing , Longnecker Richard , Connolly Sarah A.
Published: April 2015

Whereas most viruses require only a single protein to bind to and fuse with cells, herpesviruses use multiple glycoproteins to mediate virus entry, and thus communication among these proteins is required. For most alphaherpesviruses, the minimal set of viral proteins required for fusion with the host cell includes glycoproteins gD, gB, and a gH/gL heterodimer. In the current model of entry, gD binds to a cellular receptor and transmits a signal to gH/gL. This signal then triggers gB, the conserved fusion protein, to insert into the target membrane and refold to merge the viral and cellular membranes. We previously demonstrated that gB homologs from two alphaherpesviruses, herpes simplex virus 1 (HSV-1) and saimiriine herpesvirus 1 (SaHV-1), were interchangeable. In contrast, neither gD nor gH/gL functioned with heterotypic entry glycoproteins, indicating that gD and gH/gL exhibit an essential type-specific functional interaction. To map this homotypic interaction site on gH/gL, we generated HSV-1/SaHV-1 gH and gL chimeras. The functional interaction with HSV-1 gD mapped to the N-terminal domains I and II of the HSV-1 gH ectodomain. The core of HSV-1 gL that interacts with gH also was required for functional homotypic interaction. The N-terminal gH/gL domains I and II are the least conserved and may have evolved to support species-specific glycoprotein interactions.

IMPORTANCE
The first step of the herpesvirus life cycle is entry into a host cell. A coordinated interaction among multiple viral glycoproteins is required to mediate fusion of the viral envelope with the cell membrane. The details of how these glycoproteins interact to trigger fusion are unclear. By swapping the entry glycoproteins of two alphaherpesviruses (HSV-1 and SaHV-1), we previously demonstrated a functional homotypic interaction between gD and gH/gL. To define the gH and gL requirements for homotypic interaction, we evaluated the function of a panel of HSV-1/SaHV-1 gH and gL chimeras. We demonstrate that domains I and II of HSV-1 gH are sufficient to promote a functional, albeit reduced, interaction with HSV-1 gD. These findings contribute to our model of how the entry glycoproteins cooperate to mediate herpesvirus entry into the cell.