bluuebunny/crossref_metadata_2025 huggingface dataset:
Giving Girls a Shot: An Examination of Mandatory Vaccination Legislation
Authors: Doan Alesha E. , Kirkpatrick Kellee
Published: May 2013

On June 8, 2006, the Food and Drug Administration approved Merck & Co.'s vaccine Gardasil, which protects women from the human papillomavirus (HPV). Twenty‐four states began entertaining initiatives that would make the vaccine mandatory for all fifth‐ and sixth‐grade girls in public schools. However, as the vaccine gained traction in the media, the dominant issue frame put forth by Merck & Co. was undermined by several competing morality concerns raised by citizens in newspaper opinion pieces. Using an analysis of selected media coverage and a logit analysis modeling the influence of competing morality, economic, and public‐health determinants on states' decision making, we investigate the ascent of issue salience surrounding the HPV vaccine, and the policy consideration process. The results from the analysis indicate that citizen‐initiated opposition increased the salience of the topic and changed the issue framing in which the mandatory vaccine legislation was being considered. State policy consideration was influenced by a mix of morality and public‐health determinants. Moreover, Merck's attempts to influence state policy failed to increase the likelihood of policy consideration.
Campaign Promises, Democratic Governance, and Environmental Policy in the U.S. Congress
Authors: Ringquist Evan J. , Neshkova Milena I. , Aamidor Joseph
Published: May 2013

One important criterion for assessing the quality of democratic governance is the extent to which the policy process effectively translates citizen preferences into collective choices. Several scholars have observed a discrepancy between citizen preferences for strong environmental protection and weak policies adopted in the United States, indicating that the United States may fall short on this criterion. We examine one possible mechanism contributing to this discrepancy—legislator defection from campaign promises. Our data indicate that legislators in the U.S. Congress routinely defect from their campaign promises in environmental protection, undermining the link between citizen preferences and policy choice. We also find that legislators are much more likely to defect from pro‐environmental campaign promises, which moves government policy toward less stringent environmental programs. Finally, the propensity of legislators to defect from their campaign promises is systematic, with defection affected by partisanship, constituency influence, the influence of the majority party, and the likely consequences of defection for policy choice. These findings contribute empirical evidence relevant to the “mandate theory” perspective on how citizen preferences are translated into collective choices through the policy process. These findings may also complement research in comparative politics concluding that legislatures selected through single member districts adopt less stringent environmental policies than do legislatures chosen via proportional representation in that the mechanism for this effect may go through legislator defection from campaign promises.
Rivals for Influence on Counterterrorism Policy: White House Political Staff Versus Executive Branch Legal Advisors
Authors: Kassop Nancy
Published: May 2013

The Obama administration struggled throughout its first term to live up to its campaign promises to reverse the most objectionable of the Bush administration's counterterrorism policies and to govern by “rule of law” principles. It is clear that most of these policies continued with minimal changes, and some even expanded to include more controversial elements such as targeted killings. The most visible policy failure has been the inability to close the military prison at Guantanamo Bay. Efforts to accomplish this closure became entwined with two other unfulfilled campaign promises—rejection of the use of military commissions for prosecuting terrorist suspects and halting the practice of indefinite detention of uncharged suspects. In this article I analyze the impact of the role played by the rivalry for influence between White House political advisors and policy principals, on the one hand, and executive branch legal advisors, on the other, on the failure to roll back Bush's counterterrorism policies.
Improvement of the methods for skin mechanical properties evaluation through correlation between different techniques and factor analysis
Authors: Neto Patrícia , Ferreira Marta , Bahia Fernanda , Costa Paulo
Published: May 2013

Background

In the past decades, many instruments have been developed to measure skin elasticity and firmness. The offer is extensive and is constantly increasing, becoming difficult to decide which equipment and mechanical property measurement are better to portrait the desired characteristics. The aim of this study was to compare and correlate parameters assessed with different probes, based on different methodologies, to understand which probe characterizes each skin elasticity property.

Methods

Measurements were performed in the abdomen region of 34 female volunteers, with three different probes: Cutometer® SEM575, Reviscometer® RVM600 and Frictiometer® FR700. Statistical data analysis was performed by Factor Analysis on IBM SPSS Statistics 17.0.

Results

Frictiometer® and Reviscometer® measurements showed significant correlations with Cutometer's Uf, Ua, Ur and Ue parameters. Ue was strongly and significantly correlated to Uf and Ur. Ua was strongly and significantly correlated with Ur, Uf and Ue. Reviscometer® measurements were similar along the same axis, and measurements performed in a specific direction are significantly correlated with age.

Conclusion

A simpler and faster methodology for the complete assessment of elasticity and firmness of human skin is possible, in alternative to elaborate and time consuming methods, involving several equipments and parameters. The simpler methodology does not reduce the information obtained about the skin properties.
Blood screening for human immunodeficiency virus: a new algorithm to reduce the false‐positive results
Authors: Zhang R. , Sun Y. , Wang L. , Zhang K. , Xie J. , Li J.
Published: May 2013

Objectives

The objectives to this study were to evaluate the performance of an anti‐human immunodeficiency virus (HIV) blood screening test and propose a new screening algorithm for blood banks routinely using nucleic acid amplification testing (NAT) to reduce false‐positive results.

Background

Most anti‐HIV enzyme‐linked immunosorbent assay (ELISA) results are false‐positive because of the low prevalence of HIV infection and high sensitivity of the ELISAs.

Methods/Materials

A total of 281 588 voluntary donations were collected and sera reactive on one or both anti‐HIV ELISAs were confirmed by Western blot (WB) testing. All samples with nonreactive results for the two ELISAs underwent NAT. A confirmed HIV‐1‐positive result was defined by a reactive result on NAT or WB testing. Correlations between signal‐to‐cutoff ratios and the confirmed HIV‐1 infection rate were analysed for each enzyme immunoassay and two‐enzyme immunoassay combination. The positive predictive values (PPVs) of the current and proposed algorithms were calculated.

Results

Seventy‐nine donations (13·9%) were positive on WB analysis and one donation negative for anti‐HIV antibody was reactive on NAT and confirmed to be a window period donation on additional follow‐up testing. The PPV of the 567 donations reactive on one or two ELISAs was 13·9%. However, using the new screening algorithm, 457 donations underwent NAT immediately instead of WB testing. Only 110 donations were tested with WB and the PPV was 71·8%.

Conclusion

Screening for HIV is sensitive, specific and time saving for donors with this algorithm, which is suitable for HIV screening in low prevalence settings.
Global Analysis of Apicomplexan Protein S‐Acyl Transferases Reveals an Enzyme Essential for Invasion
Authors: Frénal Karine , Tay Chwen L. , Mueller Christina , Bushell Ellen S. , Jia Yonggen , Graindorge Arnault , Billker Oliver , Rayner Julian C. , Soldati‐Favre Dominique
Published: May 2013

Synopsis

Spiderweb‐like gelatine matrices generated by electrospinning are introduced as versatile substrate for culturing animal cell monolayers destined for cryofixation and EM. Thus, non‐disruptive sampling and transfer of native cells into high‐pressure freezing devices is possible within about 30 seconds. In addition to cryosection‐ and replica‐labelling, this aproach can be applied to various complementary microscopy and biochemical methods.
Cryptic diversity in South American Teiinae (Squamata, Teiidae) lizards
Authors: Giugliano Lilian G. , de Campos Nogueira Cristiano , Valdujo Paula H. , Collevatti Rosane G. , Colli Guarino R.
Published: May 2013

Based on phylogenetic and molecular dating analyses of several species of Cnemidophorus and Ameiva, representing major groups of species of these two genera, we uncover a previously unrecognized Ameiva lineage, which includes described Cnemidophorus parecis from south‐western Amazonia. We discuss the diagnosis of Ameiva and Cnemidophorus and the implications of the new taxonomic rearrangement of genera from Teiidae for the monophyly of Ameiva. Based on the conclusion of our analyses, we provide description of a new species named Ameiva jacuba from the central Brazilian Cerrado and a detailed diagnosis for the relocation of C. parecis to Ameiva. We do not adopt here recent taxonomic changes proposed for Teiidae and provide a discussion about them. Finally, based on molecular dating and the distribution of living species, we propose an evolutionary scenario for the origins of South American cis‐Andean Ameiva lineages, associated with the topographic subdivision of the Cerrado region during Miocene marine introgressions.
Demonstration of electrical connectivity between self-assembled structures
Authors: Rao Madhav , Lusth John C. , Burkett Susan L.
Published: May 2013

A novel way of three dimensional (3D) chip stacking has been designed in a view to improve heat dissipation across the layers. Solder-based self assembled (SBSA) structures have been designed as 3D posts on simulated through silicon vias to demonstrate the concept. The fabrication of SBSA structures using a low temperature solder alloy and dip soldering method is described. Previously, two types of soldering—face soldering and edge soldering—were studied to fabricate SBSA structures. Face soldering refers to deposition of solder on the complete metal face whereas edge soldering refers to selective deposition of solder on only the edges of the metal face. Mechanical grinding of the 3D structures shows that face soldered SBSA structures were void free and robust enough to be used as a connection post for chip stacking. Edge soldered SBSA structures collapsed when grinding was performed. This suggests the edge soldered 3D structure may only be partially filled. Face soldered SBSA structures provide a solder bump that serves as a connection path in the integration of dissimilar electronic technologies. Cylindrical copper posts, developed in a previous project, can be an effective approach to integrated circuit stacking. However, the SBSA post provides more variety in size and shape and can serve as a reservoir for solder to aid in chip bonding. The solder bumps are heat resistant, and uniform thicknesses were obtained across a large array of SBSA structures. The electrical durability of SBSA posts were determined by completing I-V measurements after thermal treatment. SBSA posts were subjected to thermal cycling with temperatures ranging from room temperature to 300 °C. The interconnected SBSA posts are shown to be stable until 165 °C with little variation in measured resistance.
Underwater noise from offshore oil production vessels
Authors: Erbe Christine , McCauley Robert , McPherson Craig , Gavrilov Alexander
Published: May 2013

Underwater acoustic recordings of six Floating Production Storage and Offloading (FPSO) vessels moored off Western Australia are presented. Monopole source spectra were computed for use in environmental impact assessments of underwater noise. Given that operations on the FPSOs varied over the period of recording, and were sometimes unknown, the authors present a statistical approach to noise level estimation. No significant or consistent aspect dependence was found for the six FPSOs. Noise levels did not scale with FPSO size or power. The 5th, 50th (median), and 95th percentile source levels (broadband, 20 to 2500 Hz) were 188, 181, and 173 dB re 1 μPa @ 1 m, respectively.
Attenuation as a function of the canal length of custom-molded earplugs: A pilot study
Authors: Tufts Jennifer B. , Chen Siyuan , Marshall Lynne
Published: May 2013

Custom-molded earplugs (CMEPs) whose canal segments extend beyond the second bend of the ear canal can provide excellent attenuation but can sometimes be uncomfortable. Attenuation was measured for CMEPs whose canal segments were shortened in 2-mm increments. The within-subjects design permitted illustration of the form of the function relating attenuation to canal segment length for individuals. Reduction of attenuation due to canal segment shortening was generally more pronounced for frequencies ≤1000 Hz. Some regions of the canal segments were more critical than others in maintaining attenuation. The relationship between comfort and canal segment length was not straightforward.
Improved vocal tract reconstruction and modeling using an image super-resolution technique
Authors: Zhou Xinhui , Woo Jonghye , Stone Maureen , Prince Jerry L. , Espy-Wilson Carol Y.
Published: May 2013

Magnetic resonance imaging has been widely used in speech production research. Often only one image stack (sagittal, axial, or coronal) is used for vocal tract modeling. As a result, complementary information from other available stacks is not utilized. To overcome this, a recently developed super-resolution technique was applied to integrate three orthogonal low-resolution stacks into one isotropic volume. The results on vowels show that the super-resolution volume produces better vocal tract visualization than any of the low-resolution stacks. Its derived area functions generally produce formant predictions closer to the ground truth, particularly for those formants sensitive to area perturbations at constrictions.
Controlled Flight of a Biologically Inspired, Insect-Scale Robot
Authors: Ma Kevin Y. , Chirarattananon Pakpong , Fuller Sawyer B. , Wood Robert J.
Published: May 2013

Lord of the Robotic Flies

While small-scale flying objects are ubiquitous in nature, they are quite hard to engineer. As sizes get smaller, fixed-winged flight becomes less efficient because of increased drag resistance.Maet al.(p.603) developed a tethered robotic fly with wings that flap through the use of piezo-electric materials. Control of the flight motion involved a feedback process, which allowed the tethered robotic fly to hover and make controlled flight maneuvers.
Exoplanet Habitability
Authors: Seager Sara
Published: May 2013

The search for exoplanets includes the promise to eventually find and identify habitable worlds. The thousands of known exoplanets and planet candidates are extremely diverse in terms of their masses or sizes, orbits, and host star type. The diversity extends to new kinds of planets, which are very common yet have no solar system counterparts. Even with the requirement that a planet’s surface temperature must be compatible with liquid water (because all life on Earth requires liquid water), a new emerging view is that planets very different from Earth may have the right conditions for life. The broadened possibilities will increase the future chances of discovering an inhabited world.
Emergence and Frustration of Magnetism with Variable-Range Interactions in a Quantum Simulator
Authors: Islam R. , Senko C. , Campbell W. C. , Korenblit S. , Smith J. , Lee A. , Edwards E. E. , Wang C.-C. J. , Freericks J. K. , Monroe C.
Published: May 2013

Magnetic Frustration

The study of magnetic frustration has a long history in solid-state physics, but cold-atom systems now offer the possibility of simulating the problem with exquisite control.

Islam
et al.

(p.
583
) study a system of 16 trapped ions, using the Coulomb interactions between the ions to simulate exchange interactions present in magnetic systems. The measured spin correlations provide a window into the behavior of the system, as the effective magnetic field and the range of the interactions are tuned.
Comment on “Bateman in Nature: Predation on Offspring Reduces the Potential for Sexual Selection”
Authors: Bergeron P. , Martin A. M. , Garant D. , Pelletier F.
Published: May 2013

Byers and Dunn (Reports, 9 November 2012, p. 802) claimed that predation on offspring reduced the potential for sexual selection in pronghorn. We argue that the potential for sexual selection is not affected by random offspring mortality when relative reproductive success is considered and increases when measured with the opportunity for selection, a metric that describes the potential for selection.
Response to Comments on “Bateman in Nature: Predation on Offspring Reduces the Potential for Sexual Selection”
Authors: Byers John , Dunn Stacey
Published: May 2013

Commenters objected to the way that we counted matings and offspring to calculate Bateman slopes and disagreed with our contention that predation on offspring can decrease the potential for sexual selection. We clarify what may have been misunderstandings to argue that our methods, analyses, and conclusions are correct.
Observed Properties of Extrasolar Planets
Authors: Howard Andrew W.
Published: May 2013

Observational surveys for extrasolar planets probe the diverse outcomes of planet formation and evolution. These surveys measure the frequency of planets with different masses, sizes, orbital characteristics, and host star properties. Small planets between the sizes of Earth and Neptune substantially outnumber Jupiter-sized planets. The survey measurements support the core accretion model, in which planets form by the accumulation of solids and then gas in protoplanetary disks. The diversity of exoplanetary characteristics demonstrates that most of the gross features of the solar system are one outcome in a continuum of possibilities. The most common class of planetary system detectable today consists of one or more planets approximately one to three times Earth’s size orbiting within a fraction of the Earth-Sun distance.
Thinking, Broad and Deep
Surfaces and Essences
Analogy as the Fuel and Fire of Thinking

by Douglas Hofstadter and Emmanuel Sander

Basic Books, New York, 2013. 592 pp. $35, C$38. ISBN 9780465018475.
Authors: Holyoak Keith J.
Published: May 2013

Taking analogies to be "the building blocks of thought," Hofstadter and Sander offer a theory of how the mind works that addresses cognition as a psychological (rather than neural) phenomenon.
Obey the Peptide Assembly Rules
Authors: Ardejani Maziar S. , Orner Brendan P.
Published: May 2013

Application of simple design rules enables controlled assembly of discrete peptide nanostructures.

[Also see Report by

Fletcher
et al.

]
The Pacific Swallows Fukushima's Fallout
Authors: Normile Dennis
Published: May 2013

The Tohoku disaster had one unprecedented impact: an enormous amount of radiation was deposited in the ocean.
TRPV1 Ablation Aggravates Inflammatory Responses and Organ Damage during Endotoxic Shock
Authors: Wang Youping , Wang Donna H.
Published: May 2013

To test the hypothesis that ablation of transient receptor potential vanilloid type 1 (TRPV1) channels leads to exacerbated inflammatory responses and organ damage during endotoxic shock, lipopolysaccharide (LPS; 5 million endotoxin units/kg of body weight) was injected intraperitoneally (i.p.) into wild-type (WT) and TRPV1-null mutant (TRPV1−/−) mice. Mean arterial pressure and heart rate, determined by radiotelemetry, were severely depressed after LPS injection into WT and TRPV1−/−mice, with no distinction between the two strains. At 24 h after LPS injection, renal glomerular hypercellularity and hepatocellular injury were observed in both strains, accompanying further elevated serum levels of creatinine and alanine aminotransferase in TRPV1−/−mice compared to those in WT mice. At 6 or 24 h after LPS injection, neutrophil recruitment into kidneys and livers, serum cytokine (tumor necrosis factor alpha [TNF-α], interleukin 1β [IL-1β], IL-6) and renal chemokine (KC, macrophage inflammatory protein 2 [MIP-2]) levels, and renal VCAM-1 and ICAM-1 expression were greater in TRPV1−/−mice than WT mice. In addition, increased plasma calcitonin gene-related peptide (CGRP) levels observed in WT mice 6 h after LPS injection were absent in TRPV1−/−mice. Thus, TRPV1 ablation aggravates inflammatory responses, including neutrophil infiltration, proinflammatory cytokine production, and adhesion molecule expression, leading to intensified organ damage during endotoxic shock in the absence of worsened circulatory failure. The data indicate that TRPV1 activation may attenuate endotoxin-induced organ damage, possibly via its anti-inflammatory action rather than alteration of hemodynamics.
Time Profile of Viral DNA in Aqueous Humor Samples of Patients Treated for Varicella-Zoster Virus Acute Retinal Necrosis by Use of Quantitative Real-Time PCR
Authors: Bernheim D. , Germi R. , Labetoulle M. , Romanet J. P. , Morand P. , Chiquet C.
Published: May 2013

The objective of this study was to evaluate the kinetics of varicella-zoster virus (VZV) loads using quantitative PCR (qPCR) in patients treated for acute retinal necrosis (ARN). Six patients (52 ± 13 years old) with ARN syndrome were consecutively studied. Aqueous humor (AH) was sampled from both eyes of all patients for qPCR evaluation. The patients were treated with intravenous acyclovir and intravitreal injections of antiviral drugs. The mean follow-up time was 17.6 ± 16.4 months. Main outcome measures were the numbers of viral genome copies in the AH, assessed using real-time qPCR with hydrolysis probe technology with a threshold of detection of 200 copies/ml. Two main portions of the viral load curves were observed for each patient: a plateau phase (27.8 ± 24.9 days) and a decrease in the number of viral genome copies. The mean baseline viral load was 3.4 × 10
7
± 4.45 × 10
7
copies/ml (6 × 10
6
to 1.2 × 10
8
copies/ml). The viral load decreased according to a logarithmic model, with a 50% reduction obtained in 3 ± 0.7 days. There was a significant viral load (>102 copies/ml) at 50 days after the onset of treatment, despite antiviral drugs. qPCR use demonstrated reproducible VZV DNA kinetics with a two-phase evolution: a plateau followed by a logarithmic decrease. These data suggest that high-dosage antiviral therapy administered for the conventional 10-day duration is insufficient for most patients. This series of patients responded with a similar decrease in viral load once treatment was initiated, and the data from these patients may be used to predict the responses of future patients.
Bovine Papillomavirus Type 13 DNA in Equine Sarcoids
Authors: Lunardi Michele , de Alcântara Brígida Kussumoto , Otonel Rodrigo Alejandro Arellano , Rodrigues Wagner Borges , Alfieri Alice Fernandes , Alfieri Amauri Alcindo
Published: May 2013

Equine sarcoids are locally aggressive fibroblastic neoplasms considered to be the most common skin tumors of horses worldwide. Bovine papillomavirus types 1 and 2 have typically been associated with sarcoids in equids. Investigations aiming to identify papillomavirus strains, aside from bovine papillomaviruses 1 and 2, which might be associated with sarcoid lesions, have been lacking. The aim of this article is to report the identification of a third bovine papillomavirus type, bovine papillomavirus 13, associated with equine sarcoids. Six sarcoid lesions were collected from diverse anatomical sites on two horses from southern Brazil. To detect a broad spectrum of papillomavirus strains, eight degenerate primer pairs designed to detect conserved regions on the L1 and E1 genes were tested on the DNA samples. Direct sequencing was then performed on the obtained amplicons, and sequence identities were compared with sequences from all bovine papillomavirus types. The FAP59/FAP64, MY09/MY11, and AR-E1F2/AR-E1R4 sequences generated from the sarcoids were shown to present 99 to 100% identity with bovine papillomavirus 13, a new bovine papillomavirus type previously described in cattle. The results from this study suggest that there is a need to identify bovine papillomavirus type 13 and other papillomavirus strains that might be associated with sarcoids in diverse geographical areas; such investigations might establish the frequency of occurrence of this viral type in these common tumors of equids.
Human Cryptosporidiosis Diagnosed in Western Australia: a Mixed Infection with Cryptosporidium meleagridis, the Cryptosporidium Mink Genotype, and an Unknown Cryptosporidium Species
Authors: Ng-Hublin Josephine S. Y. , Combs Barry , MacKenzie Brian , Ryan Una
Published: May 2013

This report describes a case of cryptosporidiosis from an immunocompetent patient from Perth, Western Australia, suffering from diarrhea and a spectrum of other symptoms. Molecular identification revealed that this patient was infected with three
Cryptosporidium
species—
Cryptosporidium meleagridis
, the
Cryptosporidium
mink genotype, and an unknown
Cryptosporidium
species.
Chemical Decontamination with
N
-Acetyl-
l
-Cysteine–Sodium Hydroxide Improves Recovery of Viable Mycobacterium avium subsp. paratuberculosis Organisms from Cultured Milk
Authors: Bradner L. , Robbe-Austerman S. , Beitz D. C. , Stabel J. R.
Published: May 2013

Mycobacterium avium
subsp.
paratuberculosis
is shed into the milk and feces of cows with advanced Johne's disease, allowing the transmission of
M. avium
subsp.
paratuberculosis
between animals. The objective of this study was to formulate an optimized protocol for the isolation of
M. avium
subsp.
paratuberculosis
in milk. The parameters investigated included chemical decontamination with
N
-acetyl-
l
-cysteine–sodium hydroxide (NALC-NaOH), alone and in combination with antibiotics (vancomycin, amphotericin B, and nalidixic acid), and the efficacy of solid (Herrold's egg yolk medium [HEY]) and liquid (Bactec 12B and
para
-JEM) culture media. For each experiment, raw milk samples from a known noninfected cow were inoculated with 10
2
to 10
8
CFU/ml of live
M. avium
subsp.
paratuberculosis
organisms. The results indicate that an increased length of exposure to NALC-NaOH from 5 to 30 min and an increased concentration of NaOH from 0.5 to 2.0% did not affect the viability of
M. avium
subsp.
paratuberculosis
. Additional treatment of milk samples with the antibiotics following NALC-NaOH treatment decreased the recovery of viable
M. avium
subsp.
paratuberculosis
cells more than treatment with NALC-NaOH alone. The Bactec 12B medium was the superior medium of the three evaluated for the isolation of
M. avium
subsp.
paratuberculosis
from milk, as it achieved the lowest threshold of detection. The optimal conditions for NALC-NaOH decontamination were determined to be exposure to 1.50% NaOH for 15 min followed by culture in Bactec 12B medium. This study demonstrates that chemical decontamination with NALC-NaOH resulted in a greater recovery of viable
M. avium
subsp.
paratuberculosis
cells from milk than from samples treated with hexadecylpyridinium chloride (HPC). Therefore, it is important to optimize milk decontamination protocols to ensure that low concentrations of
M. avium
subsp.
paratuberculosis
can be detected.
First Report of Nocardia asiatica Olecranon Bursitis in an Immunocompetent Traveler Returning to Austria
Authors: Leitner Eva , Valentin Thomas , Hoenigl Martin , Lanz Philipp , Flick Holger , Zollner-Schwetz Ines , Grisold Andrea J. , Feierl Gebhard , Krause Robert
Published: May 2013

Nocardia
spp. are rarely isolated in extrapulmonary clinical specimens. We describe the first case of olecranon bursitis caused by
Nocardia asiatica
. The patient, a traveler returning from Thailand, was successfully treated with linezolid.
Clinical Validation of the Abbott RealTi
m
e High Risk HPV Assay According to the Guidelines for Human Papillomavirus DNA Test Requirements for Cervical Screening
Authors: Hesselink A. T. , Meijer C. J. L. M. , Poljak M. , Berkhof J. , van Kemenade F. J. , van der Salm M. L. , Bogaarts M. , Snijders P. J. F. , Heideman D. A. M.
Published: May 2013

This study showed that the Abbott RealTi
m
e High Risk HPV assay fulfilled cross-sectional clinical equivalence and reproducibility criteria of international consensus guidelines, which indicates that this assay can be considered clinically validated for cervical cancer screening purposes.
Emergence of a New Norovirus GII.4 Variant and Changes in the Historical Biennial Pattern of Norovirus Outbreak Activity in Alberta, Canada, from 2008 to 2013
Authors: Hasing Maria E. , Lee Bonita E. , Preiksaitis Jutta K. , Tellier Raymond , Honish Lance , Senthilselvan Ambikaipakan , Pang Xiaoli L.
Published: May 2013

The public health impact of the emergence of new norovirus (NoV) strains is uncertain. A biennial pattern of alternating quiescent and epidemic levels of NoV outbreak activity associated with the emergence of new GII.4 variants was observed in Alberta, Canada, between July 2000 and June 2008. In this study, NoV genogroup I (GI) and GII strains isolated from 710 outbreak specimens in Alberta between July 2008 and January 2013 were characterized to update historical data. The seasonality and annual variation in NoV outbreak burden were analyzed over a 10-year period (July 2002 to June 2012). We found that GII.4-2006b had persisted as the predominant variant over three observation periods (July 2006 to June 2009) during which the biennial NoV outbreak pattern continued. The emergence of GII.4-2010 (winter 2009) was not associated with increased outbreak activity, and outbreak activity between July 2009 and June 2012 when GII.4-2010 predominated (67.5 to 97.7%) did not follow a biennial pattern. GII.4-2012 first emerged in Alberta in September 2011 and became predominant in observation period July 2012 to June 2013. NoV GI, relatively rare in past years, had a higher activity level (37.3%) as represented by GI.6 and GI.7 in the winter of 2012 to 2013. A higher proportion of GI outbreaks occurred in non-health care facility settings compared to GII. Our study suggests that factors other than new variants emergence contribute to the levels of NoV outbreak activity in Alberta.
Multiple-Locus Variable-Number Tandem-Repeat Analysis in Genotyping Yersinia enterocolitica Strains from Human and Porcine Origins
Authors: Virtanen S. , Laukkanen-Ninios R. , Ortiz Martínez P. , Siitonen A. , Fredriksson-Ahomaa M. , Korkeala H.
Published: May 2013

Sporadic and epidemiologically linked
Yersinia enterocolitica
strains (
n
= 379) isolated from fecal samples from human patients, tonsil or fecal samples from pigs collected at slaughterhouses, and pork samples collected at meat stores were genotyped using multiple-locus variable-number tandem-repeat analysis (MLVA) with six loci, i.e., V2A, V4, V5, V6, V7, and V9. In total, 312 different MLVA types were found. Similar types were detected (i) in fecal samples collected from human patients over 2 to 3 consecutive years, (ii) in samples from humans and pigs, and (iii) in samples from pigs that originated from the same farms. Among porcine strains, we found farm-specific MLVA profiles. Variations in the numbers of tandem repeats from one to four for variable-number tandem-repeat (VNTR) loci V2A, V5, V6, and V7 were observed within a farm. MLVA was applicable for serotypes O:3, O:5,27, and O:9 and appeared to be a highly discriminating tool for distinguishing sporadic and outbreak-related strains. With long-term use, interpretation of the results became more challenging due to variations in more-discriminating loci, as was observed for strains originating from pig farms. Additionally, we encountered unexpectedly short V2A VNTR fragments and sequenced them. According to the sequencing results, updated guidelines for interpreting V2A VNTR results were prepared.
Brachyspira Species and Gastroenteritis in Humans
Authors: Westerman L. J. , de Boer R. F. , Roelfsema J. H. , Friesema I. H. M. , Kortbeek L. M. , Wagenaar J. A. , Bonten M. J. M. , Kusters J. G.
Published: May 2013

Brachyspira
species have been implicated as a potential cause of gastroenteritis in humans; this is, however, controversial. In 733 gastroenteritis cases and 464 controls, we found 29 samples positive for
Brachyspira
species (2.3% of cases and 2.6% of controls;
P
= 0.77).
Brachyspira
species were not associated with gastroenteritis in humans.
CK2 Phosphorylation Inactivates DNA Binding by the Papillomavirus E1 and E2 Proteins
Authors: Schuck Stephen , Ruse Cristian , Stenlund Arne
Published: May 2013

Papillomaviruses have complex life cycles that are understood only superficially. Although it is well established that the viral E1 and E2 proteins play key roles in controlling viral transcription and DNA replication, how these factors are regulated is not well understood. Here, we demonstrate that phosphorylation by the protein kinase CK2 controls the biochemical activities of the bovine papillomavirus E1 and E2 proteins by modifying their DNA binding activity. Phosphorylation at multiple sites in the N-terminal domain in E1 results in the loss of sequence-specific DNA binding activity, a feature that is also conserved in human papillomavirus (HPV) E1 proteins. The bovine papillomavirus (BPV) E2 protein, when phosphorylated by CK2 on two specific sites in the hinge, also loses its site-specific DNA binding activity. Mutation of these sites in E2 results in greatly increased levels of latent viral DNA replication, indicating that CK2 phosphorylation of E2 is a negative regulator of viral DNA replication during latent viral replication. In contrast, mutation of the N-terminal phosphorylation sites in E1 has no effect on latent viral DNA replication. We propose that the phosphorylation of the N terminus of E1 plays a role only in vegetative viral DNA replication, and consistent with such a role, caspase 3 cleavage of E1, which has been shown to be necessary for vegetative viral DNA replication, restores the DNA binding activity to phosphorylated E1.
Genetic Requirement for Hemagglutinin Glycosylation and Its Implications for Influenza A H1N1 Virus Evolution
Authors: Kim Jin Il , Lee Ilseob , Park Sehee , Hwang Min-Woong , Bae Joon-Yong , Lee Sangmoo , Heo Jun , Park Mee Sook , García-Sastre Adolfo , Park Man-Seong
Published: May 2013

Influenza A virus has evolved and thrived in human populations. Since the 1918 influenza A pandemic, human H1N1 viruses had acquired additional N-linked glycosylation (NLG) sites within the globular head region of hemagglutinin (HA) until the NLG-free HA head pattern of the 1918 H1N1 virus was renewed with the swine-derived 2009 pandemic H1N1 virus. Moreover, the HA of the 2009 H1N1 virus appeared to be antigenically related to that of the 1918 H1N1 virus. Hence, it is possible that descendants of the 2009 H1N1 virus might recapitulate the acquisition of HA head glycosylation sites through their evolutionary drift as a means to evade preexisting immunity. We evaluate here the evolution signature of glycosylations found in the globular head region of H1 HA in order to determine their impact in the virulence and transmission of H1N1 viruses. We identified a polymorphism at HA residue 147 associated with the acquisition of glycosylation at residues 144 and 172. By
in vitro
and
in vivo
analyses using mutant viruses, we also found that the polymorphism at HA residue 147 compensated for the loss of replication, virulence, and transmissibility associated with the presence of the N-linked glycans. Our findings suggest that the polymorphism in H1 HA at position 147 modulates viral fitness by buffering the constraints caused by N-linked glycans and provide insights into the evolution dynamics of influenza viruses with implications in vaccine immunogenicity.
Annexin A2 and S100A10 Regulate Human Papillomavirus Type 16 Entry and Intracellular Trafficking in Human Keratinocytes
Authors: Dziduszko Agnieszka , Ozbun Michelle A.
Published: May 2013

Human papillomaviruses (HPVs) cause benign and malignant tumors of the mucosal and cutaneous epithelium. The initial events regulating HPV infection impact the establishment of viral persistence, which is requisite for malignant progression of HPV-infected lesions. However, the precise mechanisms involved in HPV entry into host cells, including the cellular factors regulating virus uptake, are not clearly defined. We show that HPV16 exposure to human keratinocytes initiates epidermal growth factor receptor (EGFR)-dependent Src protein kinase activation that results in phosphorylation and extracellular translocation of annexin A2 (AnxA2). HPV16 particles interact with AnxA2 in association with S100A10 as a heterotetramer at the cell surface in a Ca
2+
-dependent manner, and the interaction appears to involve heparan-sulfonated proteoglycans. We show multiple lines of evidence that this interaction promotes virus uptake into host cells. An antibody to AnxA2 prevents HPV16 internalization, whereas an antibody to S100A10 blocks infection at a late endosomal/lysosomal site. These results suggest that AnxA2 and S100A10 have separate roles during HPV16 binding, entry, and trafficking. Our data additionally imply that AnxA2 and S100A10 may be involved in regulating the intracellular trafficking of virus particles prior to nuclear delivery of the viral genome.
Virus-Specific Effects of TRIM5α
rh
RING Domain Functions on Restriction of Retroviruses
Authors: Li Xing , Kim Jonghwa , Song Byeongwoon , Finzi Andrés , Pacheco Beatriz , Sodroski Joseph
Published: May 2013

The tripartite motif protein TRIM5α restricts particular retrovirus infections by binding to the incoming capsid and inhibiting the early stage of virus infection. The TRIM5α RING domain exhibits E3 ubiquitin ligase activity and assists the higher-order association of TRIM5α dimers, which promotes capsid binding. We characterized a panel of RING domain mutants of the rhesus monkey TRIM5α (TRIM5α
rh
) protein. The RING domain function that significantly contributed to retroviral restriction depended upon the restricted virus. The E3 ubiquitin ligase activity of the RING domain contributes to the potency of HIV-1 restriction. Nonetheless, TRIM5α
rh
mutants without detectable E3 ubiquitin ligase activity still blocked reverse transcription and inhibited HIV-1 infection at a moderate level. When TRIM5α
rh
capsid binding was weakened by substitution with a less efficient B30.2/SPRY domain, the promotion of higher-order association by the RING domain was more important to HIV-1 restriction than its E3 ubiquitin ligase activity. For the restriction of N-tropic murine leukemia virus (N-MLV) and equine infectious anemia virus (EIAV) infection, promotion of higher-order association represented the major contribution of the RING domain. Thus, both identity of the target virus and the B30.2/SPRY domain-mediated affinity for the viral capsid determine the relative contribution of the two known RING domain functions to TRIM5α restriction of retrovirus infection.
Clustering and Mobility of HIV-1 Env at Viral Assembly Sites Predict Its Propensity To Induce Cell-Cell Fusion
Authors: Roy Nathan H. , Chan Jany , Lambelé Marie , Thali Markus
Published: May 2013

HIV-1 Env mediates virus attachment to and fusion with target cell membranes, and yet, while Env is still situated at the plasma membrane of the producer cell and before its incorporation into newly formed particles, Env already interacts with the viral receptor CD4 on target cells, thus enabling the formation of transient cell contacts that facilitate the transmission of viral particles. During this first encounter with the receptor, Env must not induce membrane fusion, as this would prevent the producer cell and the target cell from separating upon virus transmission, but how Env's fusion activity is controlled remains unclear. To gain a better understanding of the Env regulation that precedes viral transmission, we examined the nanoscale organization of Env at the surface of producer cells. Utilizing superresolution microscopy (stochasticopticalreconstructionmicroscopy [STORM]) andfluorescencerecoveryafterphotobleaching (FRAP), we quantitatively assessed the clustering and dynamics of Env upon its arrival at the plasma membrane. We found that Gag assembly induced the aggregation of small Env clusters into larger domains and that these domains were completely immobile. Truncation of the cytoplasmic tail (CT) of Env abrogated Gag's ability to induce Env clustering and restored Env mobility at assembly sites, both of which correlated with increased Env-induced fusion of infected and uninfected cells. Hence, while Env trapping by Gag secures Env incorporation into viral particles, Env clustering and its sequestration at assembly sites likely also leads to the repression of its fusion function, and thus, by preventing the formation of syncytia, Gag helps to secure efficient transfer of viral particles to target cells.
Influence of Mismatch of Env Sequences on Vaccine Protection by Live Attenuated Simian Immunodeficiency Virus
Authors: Manrique Julieta , Piatak Michael , Lauer William , Johnson Welkin , Mansfield Keith , Lifson Jeffrey , Desrosiers Ronald
Published: May 2013

Vaccine/challenge experiments that utilize live attenuated strains of simian immunodeficiency virus (SIV) in monkeys may be useful for elucidating what is needed from a vaccine in order to achieve protective immunity. Derivatives of SIVmac239 and SIVmac239Δ
nef
were constructed in which
env
sequences were replaced with those of the heterologous strain E543; these were then used in vaccine/challenge experiments. When challenge occurred at 22 weeks, 10 of 12 monkeys exhibited apparent sterilizing immunity despite a mismatch of Env sequences, compared to 12 of 12 monkeys with apparent sterilizing immunity when challenge virus was matched in its Env sequence. However, when challenge occurred at 6 weeks, 6 of 6 SIV239Δ
nef
-immunized monkeys became superinfected by challenge virus mismatched in its Env sequence (SIV239/EnvE543). These results contrast markedly not only with the results of the week 22 challenge but also with the sterilizing immunity observed in 5 of 5 SIV239Δ
nef
-immunized rhesus monkeys challenged at 5 weeks with SIV239, i.e., with no mismatch of Env sequences. We conclude from these studies that a mismatch of Env sequences in the challenge virus can have a dramatic effect on the extent of apparent sterilizing immunity when challenge occurs relatively early, 5 to 6 weeks after the
nef
-deleted SIV administration. However, by 22 weeks, mismatch of Env sequences has little or no influence on the degree of protection against challenge virus. Our findings suggest that anti-Env immune responses are a key component of the protective immunity elicited by live attenuated,
nef
-deleted SIV.
Cross-Inhibition of Chikungunya Virus Fusion and Infection by Alphavirus E1 Domain III Proteins
Authors: Sánchez-San Martín Claudia , Nanda Soumya , Zheng Yan , Fields Whitney , Kielian Margaret
Published: May 2013

Alphaviruses are small enveloped RNA viruses that include important emerging human pathogens, such as chikungunya virus (CHIKV). These viruses infect cells via a low-pH-triggered membrane fusion reaction, making this step a potential target for antiviral therapies. The E1 fusion protein inserts into the target membrane, trimerizes, and refolds to a hairpin-like conformation in which the combination of E1 domain III (DIII) and the stem region (DIII-stem) pack against a core trimer composed of E1 domains I and II (DI/II). Addition of exogenous DIII proteins from Semliki Forest virus (SFV) has been shown to inhibit E1 hairpin formation and SFV fusion and infection. Here we produced and characterized DIII and DI/II proteins from CHIKV and SFV. Unlike SFV DIII, both core trimer binding and fusion inhibition by CHIKV DIII required the stem region. CHIKV DIII-stem and SFV DIII-stem showed efficient cross-inhibition of SFV, Sindbis virus, and CHIKV infections. We developed a fluorescence anisotropy-based assay for the binding of SFV DIII-stem to the core trimer and used it to demonstrate the relatively high affinity of this interaction (

K
d

[dissociation constant], ∼85 nM) and the importance of the stem region. Together, our results support the conserved nature of the key contacts of DIII-stem in the alphavirus E1 homotrimer and describe a sensitive and quantitative
in vitro
assay for this step in fusion protein refolding.
Antiviral Activity of Trappin-2 and Elafin
In Vitro
and
In Vivo
against Genital Herpes
Authors: Drannik Anna G. , Nag Kakon , Sallenave Jean-Michel , Rosenthal Kenneth L.
Published: May 2013

Serine protease inhibitor elafin (E) and its precursor, trappin-2 (Tr), have been associated with mucosal resistance to HIV-1 infection. We recently showed that Tr/E are among principal anti-HIV-1 molecules in cervicovaginal lavage (CVL) fluid, that E is ∼130 times more potent than Tr against HIV-1, and that Tr/E inhibited HIV-1 attachment and transcytosis across human genital epithelial cells (ECs). Since herpes simplex virus 2 (HSV-2) is a major sexually transmitted infection and risk factor for HIV-1 infection and transmission, we assessed Tr/E contribution to defense against HSV-2. Our
in vitro
studies demonstrated that pretreatment of endometrial (HEC-1A) and endocervical (End1/E6E7) ECs with human Tr-expressing adenovirus (Ad/Tr) or recombinant Tr/E proteins before or after HSV-2 infection resulted in significantly reduced virus titers compared to those of controls. Interestingly, E was ∼7 times more potent against HSV-2 infection than Tr. Conversely, knockdown of endogenous Tr/E by small interfering RNA (siRNA) significantly increased HSV-2 replication in genital ECs. Recombinant Tr and E reduced viral attachment to genital ECs by acting indirectly on cells. Further, lower viral replication was associated with reduced secretion of proinflammatory interleukin 8 (IL-8) and tumor necrosis factor alpha (TNF-α) and decreased NF-κB nuclear translocation. Additionally, protected Ad/Tr-treated ECs demonstrated enhanced interferon regulatory factor 3 (IRF3) nuclear translocation and increased antiviral IFN-β in response to HSV-2. Lastly,
in vivo
studies of intravaginal HSV-2 infection in Tr-transgenic mice (Etg) showed that despite similar virus replication in the genital tract, Etg mice had reduced viral load and TNF-α in the central nervous system compared to controls. Collectively, this is the first experimental evidence highlighting anti-HSV-2 activity of Tr/E in female genital mucosa.