bluuebunny/crossref_metadata_2025 huggingface dataset:
Interfacial Effects in Iron-Nickel Hydroxide–Platinum Nanoparticles Enhance Catalytic Oxidation
Authors: Chen Guangxu , Zhao Yun , Fu Gang , Duchesne Paul N. , Gu Lin , Zheng Yanping , Weng Xuefei , Chen Mingshu , Zhang Peng , Pao Chih-Wen , Lee Jyh-Fu , Zheng Nanfeng
Published: May 2014

Improving Reactions at Interfaces

Alloying precious metals such as platinum with more abundant transition metals, such as iron and nickel, can both improve their catalytic reactivity and lower catalyst cost.

Chen
et al.

(p.
495
) explored using coatings of iron oxide–hydroxide layers on supported platinum nanoparticles for CO oxidation. The presence of this layer allowed the reaction to run rapidly at room temperature by bringing together different reaction sites on the two metals. The addition of nickel improved catalyst lifetime, and an oxidative transformation created a more complex nanoparticle morphology that increased platinum utilization.
Antibiotics and Collateral Damage
Missing Microbes
How the Overuse of Antibiotics Is Fueling Our Modern Plagues

by Martin J. Blaser

Holt, New York, 2014. 288 pp. $28. ISBN 9780805098105.
Authors: Mather Alison E.
Published: May 2014

Blaser argues that some 20th-century medical advances threaten crucial members of the human microbiome—and thus our health.
Assembly and Disassembly of Ferrocene-Based Nanotubes
Authors: Hudson Zachary M. , Manners Ian
Published: May 2014

The stacked rings of a nanotube can be dissociated and reassembled through oxidation and reduction of its constituent metal complexes.

[Also see Report by

Fukino
et al.

]
Perovskite Solar Cells Keep On Surging
Authors: Service Robert F.
Published: May 2014

The efficiency of this solar power newcomer is once again on the rise, now approaching 20%.
Drones Flying High as New Tool for Field Biologists
Authors: Schiffman Richard
Published: May 2014

Drones are becoming an essential tool for managing wildlife and fighting poachers.
Death of the Stars
Authors: Stokstad Erik
Published: May 2014

The biggest known die-off of sea stars is sweeping North America's coastal waters. Researchers are hurrying to identify the culprit and understand the impacts.
Evaluation of Immune Responses in Mice after DNA Immunization with Putative Toxoplasma gondii Calcium-Dependent Protein Kinase 5
Authors: Zhang Nian-Zhang , Huang Si-Yang , Xu Ying , Chen Jia , Wang Jin-Lei , Tian Wei-Peng , Zhu Xing-Quan
Published: May 2014

Toxoplasma gondiican cause serious public health problems and economic losses worldwide. Calcium-dependent protein kinases (CDPKs) are key mediators ofT. gondiisignaling pathways and are implicated as important virulence factors. In the present study, we cloned a novelT. gondiiCDPK gene, named TgCDPK5, and constructed the eukaryotic expression vector pVAX-CDPK5. Then, we evaluated the immune protection induced by pVAX-CDPK5 in Kunming mice. After injection of pVAX-CDPK5 intramuscularly, immune responses, determined with lymphoproliferative assays and cytokine and antibody measurements, were monitored, and mouse survival times and brain cyst formation were evaluated following challenges with theT. gondiiRH strain (genotype I) and the PRU strain (genotype II). pVAX-CDPK5 effectively induced immune responses with increased specific antibodies, a predominance of IgG2a production, and a strong lymphocyte proliferative response. The levels of gamma interferon (IFN-γ), interleukin 2 (IL-2), and IL-12(p70) and the percentages of CD3+CD4+and CD3+CD8+cells in mice vaccinated with pVAX-CDPK5 were significantly increased. However, IL-4 and IL-10 were not produced in the vaccinated mice. These results demonstrate that pVAX-CDPK5 can elicit strong humoral and cellular Th1 immune responses. The survival time of immunized mice challenged with theT. gondiiRH strain (8.67 ± 4.34 days) was slightly, but not significantly, longer than that in the control groups within 7 days (P> 0.05). The numbers of brain cysts in the mice in the pVAX-CDPK5 group were reduced by ∼40% compared with those in the control groups (P< 0.05), which provides a foundation for the further development of effective subunit vaccines againstT. gondii.
Antibodies for Treatment of Clostridium difficile Infection
Authors: Humphreys David P. , Wilcox Mark H.
Published: May 2014

Antibodies for the treatment ofClostridium difficileinfection (CDI) have been demonstrated to be effective in the research and clinical environments. Early uncertainties about molecular and treatment modalities now appear to have converged upon the systemic dosing of mixtures of human IgG1. Although multiple examples of high-potency monoclonal antibodies (MAbs) exist, significant difficulties were initially encountered in their discovery. This minireview describes historical and contemporary MAbs and highlights differences between the most potent MAbs, which may offer insight into the pathogenesis and treatment of CDI.
Production and Preclinical Evaluation of Plasmodium falciparum MSP-1
19
and MSP-3
11
Chimeric Protein, PfMSP-Fu
24
Authors: Gupta Puneet K. , Mukherjee Paushali , Dhawan Shikha , Pandey Alok K. , Mazumdar Suman , Gaur Deepak , Jain S. K. , Chauhan Virander S.
Published: May 2014

A
Plasmodium falciparum
chimeric protein, PfMSP-Fu
24
, was constructed by genetically coupling immunodominant, conserved regions of two merozoite surface proteins, the 19-kDa region C-terminal region of merozoite surface protein 1 (PfMSP-1
19
) and an 11-kDa conserved region of merozoite surface protein 3 (PfMSP-3
11
), to augment the immunogenicity potential of these blood-stage malaria vaccine candidates. Here we describe an improved, efficient, and scalable process to produce high-quality PfMSP-Fu
24
. The chimeric protein was produced in
Escherichia coli
SHuffle T7 Express
lysY
cells that express disulfide isomerase DsbC. A two-step purification process comprising metal affinity followed by cation exchange chromatography was developed, and we were able to obtain PfMSP-Fu
24
with purity above 99% and with a considerable yield of 23 mg/liter. Immunogenicity of PfMSP-Fu
24
formulated with several adjuvants, including Adjuplex, Alhydrogel, Adjuphos, Alhydrogel plus glucopyranosyl lipid adjuvant, aqueous (GLA-AF), Adjuphos+GLA-AF, glucopyranosyl lipid adjuvant-stable emulsion (GLA-SE), and Freund's adjuvant, was evaluated. PfMSP-Fu
24
formulated with GLA-SE and Freund's adjuvant in mice and with Alhydrogel and Freund's adjuvant in rabbits produced high titers of PfMSP-1
19
and PfMSP-3
11
-specific functional antibodies. Some of the adjuvant formulations induced inhibitory antibody responses and inhibited
in vitro
growth of
P. falciparum
parasites in the presence as well as in the absence of human monocytes. These results suggest that PfMSP-Fu
24
can form a constituent of a multistage malaria vaccine.
Performance of the OraQuick HCV Rapid Antibody Test for Screening Exposed Patients in a Hepatitis C Outbreak Investigation
Authors: Gao Fengxiang , Talbot Elizabeth A. , Loring Carol H. , Power Jill J. , Dionne-Odom Jodie , Alroy-Preis Sharon , Jackson Patricia , Bean Christine L.
Published: May 2014

During a nosocomial hepatitis C outbreak, emergency public clinics employed the OraQuick HCV rapid antibody test on site, and all results were verified by a standard enzyme immunoassay (EIA). Of 1,157 persons, 1,149 (99.3%) exhibited concordant results between the two tests (16 positive, 1,133 negative). The sensitivity, specificity, positive predictive value, and negative predictive value were 94.1%, 99.5%, 72.7%, and 99.9%, respectively. OraQuick performed well as a screening test during an outbreak investigation and could be integrated into future hepatitis C virus (HCV) outbreak testing algorithms.
Next-Generation Sequencing for Typing and Detection of Resistance Genes: Performance of a New Commercial Method during an Outbreak of Extended-Spectrum-Beta-Lactamase-Producing Escherichia coli
Authors: Veenemans J. , Overdevest I. T. , Snelders E. , Willemsen I. , Hendriks Y. , Adesokan A. , Doran G. , Bruso S. , Rolfe A. , Pettersson A. , Kluytmans J. A. J. W.
Published: May 2014

Next-generation sequencing (NGS) has the potential to provide typing results and detect resistance genes in a single assay, thus guiding timely treatment decisions and allowing rapid tracking of transmission of resistant clones. We evaluated the performance of a new NGS assay (Hospital Acquired Infection BioDetection System; Pathogenica) during an outbreak of sequence type 131 (ST131)
Escherichia coli
infections in a nursing home in The Netherlands. The assay was performed on 56 extended-spectrum-beta-lactamase (ESBL)
E. coli
isolates collected during 2 prevalence surveys (March and May 2013). Typing results were compared to those of amplified fragment length polymorphism (AFLP), whereby we visually assessed the agreement of the BioDetection phylogenetic tree with clusters defined by AFLP. A microarray was considered the gold standard for detection of resistance genes. AFLP identified a large cluster of 31 indistinguishable isolates on adjacent departments, indicating clonal spread. The BioDetection phylogenetic tree showed that all isolates of this outbreak cluster were strongly related, while the further arrangement of the tree also largely agreed with other clusters defined by AFLP. The BioDetection assay detected ESBL genes in all but 1 isolate (sensitivity, 98%) but was unable to discriminate between ESBL and non-ESBL TEM and SHV beta-lactamases or to specify CTX-M genes by group. The performance of the hospital-acquired infection (HAI) BioDetection System for typing of
E. coli
isolates compared well with the results of AFLP. Its performance with larger collections from different locations, and for typing of other species, was not evaluated and needs further study.
Evaluation of Curetis Unyvero, a Multiplex PCR-Based Testing System, for Rapid Detection of Bacteria and Antibiotic Resistance and Impact of the Assay on Management of Severe Nosocomial Pneumonia
Authors: Jamal Wafaa , Al Roomi Ebtehal , AbdulAziz Lubna R. , Rotimi Vincent O.
Published: May 2014

Health care-associated pneumonia due to multidrug-resistant organisms represents a major therapeutic challenge. Unfortunately, treatment is dependent on empirical therapy, which often leads to improper and inadequate antimicrobial therapy. A rapid multiplex PCR-based Unyvero pneumonia application (UPA) assay that assists in timely decision-making has recently become available. In this study, we evaluated the performance of UPA in detecting etiological pathogens and resistance markers in patients with nosocomial pneumonia (NP). The impact of this assay on the management of severe nosocomial pneumonia was also assessed. Appropriate specimens were processed by UPA according to the manufacturer's protocol in parallel with conventional culture methods. Of the 56 patients recruited into the study, 49 (87.5%) were evaluable. Of these, 27 (55.1%) and 4 (8.2%) harbored multiple bacteria by the PCR assay and conventional culture, respectively. A single pathogen was detected in 8 (16.3%) and 4 (8.2%) patients, respectively. Thirteen different genes were detected from 38 patients, including the
ermB
gene (40.8%), the
bla
OXA-51
-like gene (28.6%), the
sul1
(28.6%) and
int1
(20.4%) integrase genes, and the
mecA
and
bla
CTX-M
genes (12.3% each). The time from sample testing to results was 4 h versus 48 to 96 h by UPA and culture, respectively. Initial empirical treatment was changed within 5 to 6 h in 33 (67.3%) patients based on the availability of UPA results. Thirty (62.2%) of the patients improved clinically. A total of 3 (6.1%) patients died, mainly from their comorbidities. These data demonstrate the potential of a multiplex PCR-based assay for accurate and timely detection of etiological agents of NP, multidrug-resistant (MDR) organisms, and resistance markers, which can guide clinicians in making early antibiotic adjustments.
Development and Clinical Evaluation of
sdaA
Loop-Mediated Isothermal Amplification Assay for Detection of Mycobacterium tuberculosis with an Approach To Prevent Carryover Contamination
Authors: Nimesh Manoj , Joon Deepali , Varma-Basil Mandira , Saluja Daman
Published: May 2014

A rapid and sensitive loop-mediated isothermal amplification assay for the
sdaA
gene of
Mycobacterium tuberculosis
was developed using a dUTP-uracil-
N
-glycosylase (dUTP-UNG) strategy to prevent carryover contamination. Evaluation of the assay using clinical specimens (
n
= 648) showed high specificity (97.2%) and sensitivity (100%), demonstrating its potential as a diagnostic test for tuberculosis, especially in resource-limited settings.
Predicted Configurations of Oligosaccharide Extensions in the Lipooligosaccharide of Nontypeable Haemophilus influenzae Isolates
Authors: McCrea Kirk W. , Xie Jingping , Daniel Deborah , Ulrich-Lewis Justin Theophilus , Zhang Lixin
Published: May 2014

Lipooligosaccharide configurations were predicted in nontypeable
Haemophilus influenzae
isolates based on the presence of seven oligosaccharide extension-initiating genes (or alleles). Predicted configurations with 2 to 3 oligosaccharide extensions were more prevalent among middle ear than throat strains. In addition, strains with these configurations averaged higher levels of serum resistance than strains with other configurations.
Mixed Mycobacterium tuberculosis Complex Infections and False-Negative Results for Rifampin Resistance by GeneXpert MTB/RIF Are Associated with Poor Clinical Outcomes
Authors: Zetola Nicola M. , Shin Sanghyuk S. , Tumedi Kefentse A. , Moeti Keletso , Ncube Ronald , Nicol Mark , Collman Ronald G. , Klausner Jeffrey D. , Modongo Chawangwa
Published: May 2014

The Xpert MTB/RIF (Xpert) assay is becoming a principal screening tool for diagnosing rifampin-resistant
Mycobacterium tuberculosis
complex (MTBC) infection. However, little is known about the performance of the Xpert assay in infections with both drug-sensitive and drug-resistant strains (mixed MTBC infections). We assessed the performance of the Xpert assay for detecting rifampin resistance using phenotypic drug sensitivity testing (DST) as the reference standard in 370 patients with microbiologically proven pulmonary tuberculosis. Mixed MTBC infections were identified genetically through 24-locus mycobacterial interspersed repetitive-unit–variable-number tandem-repeat (MIRU-VNTR) analysis. Logistic regression was used to identify the factors associated with poor (defined as treatment failure, default, and death from any cause) or good (defined as cure or successful treatment completion) clinical outcomes. The analytic sensitivity of the Xpert assay for detecting rifampin resistance was assessed
in vitro
by testing cultures containing different ratios of drug-sensitive and drug-resistant organisms. Rifampin resistance was detected by the Xpert assay in 52 (14.1%) and by phenotypic DST in 55 (14.9%) patients. Mixed MTBC infections were identified in 37 (10.0%) patients. The Xpert assay was 92.7% (95% confidence interval [CI], 82.4% to 97.9%) sensitive for detecting rifampin resistance and 99.7% (95% CI, 98.3% to 99.9%) specific. When restricted to patients with mixed MTBC infections, Xpert sensitivity was 80.0% (95% CI, 56.3 to 94.3%). False-negative Xpert results (adjusted odds ratio [aOR], 6.6; 95% CI,1.2 to 48.2) and mixed MTBC infections (aOR, 6.5; 95% CI, 2.1 to 20.5) were strongly associated with poor clinical outcome. The Xpert assay failed to detect rifampin resistance
in vitro
when <90% of the organisms in the sample were rifampin resistant. Our study indicates that the Xpert assay has an increased false-negative rate for detecting rifampin resistance with mixed MTBC infections. In hyperendemic settings where mixed infections are common, the Xpert results might need further confirmation.
Multicenter Feasibility Study To Assess External Quality Assessment Panels for Xpert MTB/RIF Assay in South Africa
Authors: Scott Lesley , Albert Heidi , Gilpin Chris , Alexander Heather , DeGruy Kyle , Stevens Wendy
Published: May 2014

External quality assessment (EQA) for the Xpert MTB/RIF assay is part of the quality system required for clinical and laboratory practice. Five newly developed EQA panels that use different matrices, including a lyophilized sample (Vircell, Granada, Spain), a dried tube specimen (CDC), liquid (Maine Molecular Quality Control, Inc. [MMQCI], Scarborough, ME), artificial sputum (Global Laboratory Initiative [GLI]), and a dried culture spot (National Health Laboratory Services [NHLS]), were evaluated at 11 GeneXpert testing sites in South Africa. The panels comprised
Mycobacterium tuberculosis
complex (MTBC)-negative, MTBC-positive (including rifampin [RIF] susceptible and RIF resistant), and nontuberculosis mycobacterial material that was inactivated and safe for transportation. Twelve qualitative and quantitative variables were scored as acceptable (1) or unacceptable (0); the overall panel performance score for the Vircell, CDC, GLI, and NHLS panels was 9 of 12, while the MMQCI panel scored 6 of 12 (owing to the need for cold chain maintenance). All panels showed good compatibility with Xpert MTB/RIF testing, and none showed PCR inhibition. The use of a liquid or dry matrix did not appear to be a distinguishing criterion, as both matrices had reduced scores on insufficient volumes, a need for extra consumables, and the ability to transfer to the Xpert MTB/RIF cartridge. EQA is an important component of the quality system required for diagnostic testing programs, but it must be complemented by routine monitoring of performance indicators and instrument verification. This study aims to introduce EQA concepts for Xpert MTB/RIF testing and evaluates five potential EQA panels.
Capture of syncytin-Mar1, a Fusogenic Endogenous Retroviral Envelope Gene Involved in Placentation in the Rodentia Squirrel-Related Clade
Authors: Redelsperger François , Cornelis Guillaume , Vernochet Cécile , Tennant Bud C. , Catzeflis François , Mulot Baptiste , Heidmann Odile , Heidmann Thierry , Dupressoir Anne
Published: May 2014

Syncytin
genes are fusogenic envelope protein (
env
) genes of retroviral origin that have been captured for a function in placentation. Within rodents, two such genes have previously been identified in the mouse-related clade, allowing a demonstration of their essential role via knockout mice. Here, we searched for similar genes in a second major clade of the Rodentia order, the squirrel-related clade, taking advantage of the complete sequencing of the ground squirrel
Ictidomys tridecemlineatus
genome.
In silico
search for
env
genes with full coding capacity identified several candidate genes with one displaying placenta-specific expression, as revealed by quantitative reverse transcription-PCR analysis of a large panel of tissues. This gene belongs to a degenerate endogenous retroviral element, with recognizable hallmarks of an integrated provirus. Cloning of the gene in an expression vector for
ex vivo
cell-cell fusion and pseudotype assays demonstrated fusogenicity on a large panel of mammalian cells.
In situ
hybridization on placenta sections showed specific expression in domains where trophoblast cells fuse into a syncytiotrophoblast at the fetomaternal interface, consistent with a role in syncytium formation. Finally, we show that the gene is conserved among the tribe Marmotini, thus dating its capture back to about at least 25 million years ago, with evidence for purifying selection and conservation of fusogenic activity. This gene that we named
syncytin-Mar1
is distinct from all seven
Syncytin
genes identified to date in eutherian mammals and is likely to be a major effector of placentation in its related clade.

IMPORTANCE
Syncytin
genes are fusogenic envelope genes of retroviral origin, ancestrally captured for a function in placentation. Within rodents, two such genes had been previously identified in the mouse-related clade. Here, in the squirrel-related rodent clade, we identified the envelope gene of an endogenous retrovirus with all the features of a
Syncytin
: it is specifically expressed in the placenta of the woodchuck
Marmota monax
, at the level of cells fusing into a syncytium; it can trigger cell-cell and virus-cell fusion
ex vivo
; and it has been conserved for >25 million years of evolution, suggesting an essential role in its host physiology. Remarkably,
syncytin-Mar1
is unrelated to all other
Syncytin
genes identified thus far in mammals (primates, muroids, carnivores, and ruminants). These results extend the range of retroviral envelope gene “domestication” in mammals and show that these events occurred independently, on multiple occasions during evolution to improve placental development in a process of convergent evolution.
Specificities of Human CD4
+
T Cell Responses to an Inactivated Flavivirus Vaccine and Infection: Correlation with Structure and Epitope Prediction
Authors: Schwaiger Julia , Aberle Judith H. , Stiasny Karin , Knapp Bernhard , Schreiner Wolfgang , Fae Ingrid , Fischer Gottfried , Scheinost Ondrej , Chmelik Vaclav , Heinz Franz X.
Published: May 2014

Tick-borne encephalitis (TBE) virus is endemic in large parts of Europe and Central and Eastern Asia and causes more than 10,000 annual cases of neurological disease in humans. It is closely related to the mosquito-borne yellow fever, dengue, Japanese encephalitis, and West Nile viruses, and vaccination with an inactivated whole-virus vaccine can effectively prevent clinical disease. Neutralizing antibodies are directed to the viral envelope protein (E) and an accepted correlate of immunity. However, data on the specificities of CD4
+
T cells that recognize epitopes in the viral structural proteins and thus can provide direct help to the B cells producing E-specific antibodies are lacking. We therefore conducted a study on the CD4
+
T cell response against the virion proteins in vaccinated people in comparison to TBE patients. The data obtained with overlapping peptides in interleukin-2 (IL-2) enzyme-linked immunosorbent spot (ELISpot) assays were analyzed in relation to the three-dimensional structures of the capsid (C) and E proteins as well as to epitope predictions based on major histocompatibility complex (MHC) class II peptide affinities. In the C protein, peptides corresponding to two out of four alpha helices dominated the response in both vaccinees and patients, whereas in the E protein concordance of immunodominance was restricted to peptides of a single domain (domain III). Epitope predictions were much better for C than for E and were especially erroneous for the transmembrane regions. Our data provide evidence for a strong impact of protein structural features that influence peptide processing, contributing to the discrepancies observed between experimentally determined and computer-predicted CD4
+
T cell epitopes.

IMPORTANCE
Tick-borne encephalitis virus is endemic in large parts of Europe and Asia and causes more than 10,000 annual cases of neurological disease in humans. It is closely related to yellow fever, dengue, Japanese encephalitis, and West Nile viruses, and vaccination with an inactivated vaccine can effectively prevent disease. Both vaccination and natural infection induce the formation of antibodies to a viral surface protein that neutralize the infectivity of the virus and mediate protection. B lymphocytes synthesizing these antibodies require help from other lymphocytes (helper T cells) which recognize small peptides derived from proteins contained in the viral particle. Which of these peptides dominate immune responses to vaccination and infection, however, was unknown. In our study we demonstrate which parts of the proteins contribute most strongly to the helper T cell response, highlight specific weaknesses of currently available approaches for their prediction, and demonstrate similarities and differences between vaccination and infection.
Investigating the Role of F-Actin in Human Immunodeficiency Virus Assembly by Live-Cell Microscopy
Authors: Rahman Sheikh Abdul , Koch Peter , Weichsel Julian , Godinez William J. , Schwarz Ulrich , Rohr Karl , Lamb Don C. , Kräusslich Hans-Georg , Müller Barbara
Published: May 2014

Human immunodeficiency virus type 1 (HIV-1) particles assemble at the plasma membrane, which is lined by a dense network of filamentous actin (F-actin). Large amounts of actin have been detected in HIV-1 virions, proposed to be incorporated by interactions with the nucleocapsid domain of the viral polyprotein Gag. Previous studies addressing the role of F-actin in HIV-1 particle formation using F-actin-interfering drugs did not yield consistent results. Filamentous structures pointing toward nascent HIV-1 budding sites, detected by cryo-electron tomography and atomic force microscopy, prompted us to revisit the role of F-actin in HIV-1 assembly by live-cell microscopy. HeLa cells coexpressing HIV-1 carrying fluorescently labeled Gag and a labeled F-actin-binding peptide were imaged by live-cell total internal reflection fluorescence microscopy (TIR-FM). Computational analysis of image series did not reveal characteristic patterns of F-actin in the vicinity of viral budding sites. Furthermore, no transient recruitment of F-actin during bud formation was detected by monitoring fluorescence intensity changes at nascent HIV-1 assembly sites. The chosen approach allowed us to measure the effect of F-actin-interfering drugs on the assembly of individual virions in parallel with monitoring changes in the F-actin network of the respective cell. Treatment of cells with latrunculin did not affect the efficiency and dynamics of Gag assembly under conditions resulting in the disruption of F-actin filaments. Normal assembly rates were also observed upon transient stabilization of F-actin by short-term treatment with jasplakinolide. Taken together, these findings indicate that actin filament dynamics are dispensable for HIV-1 Gag assembly at the plasma membrane of HeLa cells.

IMPORTANCE
HIV-1 particles assemble at the plasma membrane of virus-producing cells. This membrane is lined by a dense network of actin filaments that might either present a physical obstacle to the formation of virus particles or generate force promoting the assembly process. Drug-mediated interference with the actin cytoskeleton showed different results for the formation of retroviral particles in different studies, likely due to general effects on the cell upon prolonged drug treatment. Here, we characterized the effect of actin-interfering compounds on the HIV-1 assembly process by direct observation of virus formation in live cells, which allowed us to measure assembly rate constants directly upon drug addition. Virus assembly proceeded with normal rates when actin filaments were either disrupted or stabilized. Taken together with the absence of characteristic actin filament patterns at viral budding sites in our analyses, this indicates that the actin network is dispensable for HIV-1 assembly.
Repertoire of Epitopes Recognized by Serum IgG from Humans Vaccinated with Herpes Simplex Virus 2 Glycoprotein D
Authors: Whitbeck J. Charles , Huang Zhen-Yu , Cairns Tina M. , Gallagher John R. , Lou Huan , Ponce-de-Leon Manuel , Belshe Robert B. , Eisenberg Roselyn J. , Cohen Gary H.
Published: May 2014

The results of a clinical trial of a subunit vaccine against genital herpes were recently reported (R. B. Belshe, P. A. Leone, D. I. Bernstein, A. Wald, M. J. Levin, J. T. Stapleton, I. Gorfinkel, R. L. Morrow, M. G. Ewell, A. Stokes-Riner, G. Dubin, T. C. Heineman, J. M. Schulte, C. D. Deal, N. Engl. J. Med.
366:
34–43, 2012, doi:10.1056/NEJMoa1103151). The vaccine consisted of a soluble form of herpes simplex virus 2 (HSV-2) glycoprotein D (gD2) with adjuvant. The goal of the current study was to examine the composition of the humoral response to gD2 within a selected subset of vaccinated individuals. Serum samples from 30 vaccine recipients were selected based upon relative enzyme-linked immunosorbent assay (ELISA) titers against gD2; 10 samples had high titers, 10 had medium titers, and the remaining 10 had low ELISA titers. We employed a novel, biosensor-based monoclonal antibody (MAb)-blocking assay to determine whether gD2 vaccination elicited IgG responses against epitopes overlapping those of well-characterized MAbs. Importantly, IgGs from the majority of gD2-immunized subjects competed for gD binding with four antigenically distinct virus-neutralizing MAbs (MC2, MC5, MC23, and DL11). Screening of patient IgGs against overlapping peptides spanning the gD2 ectodomain revealed that about half of the samples contained antibodies against linear epitopes within the N and C termini of gD2. We found that the virus-neutralizing abilities of the 10 most potent samples correlated with overall gD-binding activity and to an even greater extent with the combined content of IgGs against the epitopes of MAbs MC2, MC5, MC23, and DL11. This suggests that optimal virus-neutralizing activity is achieved by strong and balanced responses to the four major discontinuous neutralizing epitopes of gD2.

IMPORTANCE
Several herpes simplex virus 2 (HSV-2) subunit vaccine studies have been conducted in human subjects using a recombinant form of HSV-2 glycoprotein D (gD2). Although several distinct, well-characterized virus-neutralizing epitopes on gD2 are targeted by murine monoclonal antibodies, it is not known whether the same epitopes are targeted by the humoral response to gD2 in humans. We have developed a novel, biosensor-based competition assay to directly address this important question. Using this approach, we identified epitopes that elicit strong humoral responses in humans, as well as other epitopes that elicit much weaker responses. These data provide new insight into the human response to known neutralizing gD2 epitopes and reveal characteristics of this response that may guide future vaccine development.
A Small Molecule Inhibits Virion Attachment to Heparan Sulfate- or Sialic Acid-Containing Glycans
Authors: Colpitts Che C. , Schang Luis M.
Published: May 2014

Primary attachment to cellular glycans is a critical entry step for most human viruses. Some viruses, such as herpes simplex virus type 1 (HSV-1) and hepatitis C virus (HCV), bind to heparan sulfate, whereas others, such as influenza A virus (IAV), bind to sialic acid. Receptor mimetics that interfere with these interactions are active against viruses that bind to either heparan sulfate or to sialic acid. However, no molecule that inhibits the attachment of viruses in both groups has yet been identified. Epigallocatechin gallate (EGCG), a green tea catechin, is active against many unrelated viruses, including several that bind to heparan sulfate or to sialic acid. We sought to identify the basis for the broad-spectrum activity of EGCG. Here, we show that EGCG inhibits the infectivity of a diverse group of enveloped and nonenveloped human viruses. EGCG acts directly on the virions, without affecting the fluidity or integrity of the virion envelopes. Instead, EGCG interacts with virion surface proteins to inhibit the attachment of HSV-1, HCV, IAV, vaccinia virus, adenovirus, reovirus, and vesicular stomatitis virus (VSV) virions. We further show that EGCG competes with heparan sulfate for binding of HSV-1 and HCV virions and with sialic acid for binding of IAV virions. Therefore, EGCG inhibits unrelated viruses by a common mechanism. Most importantly, we have identified EGCG as the first broad-spectrum attachment inhibitor. Our results open the possibility for the development of small molecule broad-spectrum antivirals targeting virion attachment.

IMPORTANCEThis study shows that it is possible to develop a small molecule antiviral or microbicide active against the two largest groups of human viruses: those that bind to glycosaminoglycans and those that bind to sialoglycans. This group includes the vast majority of human viruses, including herpes simplex viruses, cytomegalovirus, influenza virus, poxvirus, hepatitis C virus, HIV, and many others.
Frequency and Abundance of Alphaherpesvirus DNA in Human Thoracic Sympathetic Ganglia
Authors: Nagel Maria A. , Rempel April , Huntington Jonathon , Kim Forrest , Choe Alexander , Gilden Don
Published: May 2014

Alphaherpesvirus reactivation from thoracic sympathetic ganglia (TSG) and transaxonal spread to target organs cause human visceral disease. Yet alphaherpesvirus latency in TSG has not been well characterized. In this study, quantitative PCR detected varicella-zoster virus (VZV), herpes simplex virus 1 (HSV-1), and HSV-2 DNA in 117 fresh TSG obtained postmortem from 15 subjects. VZV DNA was found in 76 (65%) ganglia from all subjects, HSV-1 DNA was found in 5 (4%) ganglia from 3 subjects, and no HSV-2 was found.
A Community Jury on PSA screening: what do well-informed men want the government to do about prostate cancer screening—a qualitative analysis
Authors: Rychetnik Lucie , Doust Jenny , Thomas Rae , Gardiner Robert , MacKenzie Geraldine , Glasziou Paul
Published: May 2014

Objective

Cancer screening policies and programmes should take account of public values and concerns. This study sought to determine the priorities, values and concerns of men who were ‘fully informed’ about the benefits and harms of prostate-specific antigen (PSA) screening; and empirically examine the value of a community jury in eliciting public values on PSA screening.

Setting

Community jury was convened on the Gold Coast, Queensland (Australia) to consider PSA screening benefits and harms, and whether government campaigns on PSA screening should be conducted.

Participants

27 men (volunteers) aged 50–70 with no personal history of prostate cancer and willing to attend jury 6–7 April 2013: 12 were randomly allocated to jury (11 attended).

Outcome measures

A qualitative analysis was conducted of the jury deliberations (audio-recorded and transcribed) to elicit the jury's views and recommendations. A survey determined the impact of the jury process on participants’ individual testing decisions compared with control group.

Results

The jury concluded governments should not invest in programmes focused on PSA screening directed at the public because the PSA test did not offer sufficient reassurance or benefit and could raise unnecessary alarm. It recommended an alternative programme to support general practitioners to provide patients with better quality and more consistent information about PSA screening. After the jury, participants were less likely to be tested in the future compared with the controls, but around half said they would still consider doing so.

Conclusions

The jury's unanimous verdict about government programmes was notable in the light of their divergent views on whether or not they would be screened themselves in the future. Community juries provide valuable insights into the priorities and concerns of men weighing up the benefits and harms of PSA screening. It will be important to assess the degree to which the findings are generalisable to other settings.
Deployed research
Authors: Nordmann Giles , Woolley T , Doughty H , Dalle Lucca J , Hutchings S , Kirkman E
Published: May 2014

The Role 3 Medical Treatment Facility (Field Hospital) in Camp Bastion (R3 Bastion) is acknowledged to be one of the busiest dedicated trauma facilities in the world. Casualties typically present with severe injuries and in physiological extremis. These casualties form a unique cohort representing the most relevant population to evaluate the effectiveness of treating battlefield injuries as academic clinicians and scientists interested in trauma seek to improve outcomes for such patients in the future. This article describes four separate but related research projects that have been undertaken in Camp Bastion, Afghanistan, over the last year. They traverse the spectrum of clinical research, ranging from data collection to a randomised control trial. The aim is to discuss some of the problems encountered and the solutions that made it possible to undertake research in a theatre of operations, thereby providing a starting point for others who may wish to initiate research in a similar environment.
ABSTRACTS FROM THE TRI-SERVICE EMERGENCY MEDICINE CONFERENCE, 2013
Authors: Smith Jason E
Published: May 2014

The 2013 Tri-Service Emergency Medicine Conference was held in the Defence CBRN Centre, Winterbourne Gunner, from 14–16 October 2013. Abstracts were invited from military trainees in emergency medicine, and subjected to standardised assessment and scoring. The authors of the four highest scoring abstracts were invited to present their abstracts orally in the final conference session.

The top four scoring abstracts, that were included in the oral presentation session, are included below.
Coxiella burnetii seroprevalence and risk factors on commercial sheep farms in The Netherlands
Authors: Schimmer B. , de Lange M. M. A. , Hautvast J. L. A. , Vellema P. , van Duynhoven Y. T. H. P.
Published: May 2014

Coxiella burnetii seroprevalence was assessed on Dutch dairy and non‐dairy sheep farms using ELISA. Risk factors for seropositivity on non‐dairy sheep farms were identified at farm and sheep level by univariate and multivariate multilevel analyses. Based on 953 dairy and 5671 non‐dairy serum samples, sheep seroprevalences were 18.7 per cent and 2.0 per cent, respectively, and 78.6 per cent and 30.5 per cent at farm level. Significant risk factors for non‐dairy sheep farms were farm location in the south of the country, sheep kept on marginal grounds, one or several supply addresses for ewes during 2007–2009 and wearing farm boots and/or outfit by professional visitors. On sheep level, risk factors included among others farm location in the south of the country, lamb breeding as main farm purpose, goat density within 10 km farm radius, use of windbreak curtain or windshields, and presence of ≥6 stillborn lambs in 2009. Farm location in the south of the country and goat density suggests that infected goats have played a role in the transmission to non‐dairy sheep. Other risk factors suggest introduction of the bacterium through sheep supply and professional visitors. Biosecurity measures should be strengthened, including avoiding infection during handling of stillborn lambs and birth products in the lambing period.
Plasma 25-hydroxyvitamin D, more so than its epimer, has a linear relationship to leaner body composition across infancy in healthy term infants
Authors: Hazell Tom J. , Gallo Sina , Berzina llze , Vanstone Catherine A. , Rodd Celia , Weiler Hope A.
Published: May 2014

Vitamin D status positively associates with skeletal muscle mass and function in adolescents. The C-3 alpha epimer of 25-hydroxyvitamin D3 (3-epi-25(OH)D3) is high in infants, yet the potential impacts of 25-hydroxyvitamin D3 (25(OH)D3) and 3-epi-25(OH)D3 on skeletal muscle development are largely unexplored. The objective of this study was (i) to explore how the concentrations of 25(OH)D3 and 3-epi-25(OH)D3 track with body composition (lean mass (LM) and fat mass (FM)) and (ii) to determine the association between 25(OH)D3 and 3-epi-25(OH)D3 in infancy. Healthy breastfed infants (n = 132) were followed from 1 to 12 months of age as part of a vitamin D dose–response study (NCT00381914). Anthropometry and diet were assessed. Body composition was measured with dual-energy X-ray absorptiometry. Plasma 25(OH)D3 and 3-epi-25(OH)D3 concentrations were evaluated using liquid chromatography tandem mass spectrometry. Plasma 25(OH)D3 and 3-epi-25(OH)D3 increased from 1 to 3 months of age and decreased thereafter (p < 0.05). Infants with 25(OH)D3 concentrations above 75 nmol/L did not have a higher LM (g or %; p > 0.273) than those below this cutoff. LM was not associated with 25(OH)D3, whereas LM% was positively associated with 25(OH)D3 (β = 0.03; CI: 0.01 to 0.06; p = 0.006), while accounting for sex, weight-for-age Z-score, protein and fat intake, and age. For FM, the variables accounting for a significant amount of the variation were plasma 25(OH)D3 concentration (β = −2.38; CI: −4.35, −0.41; p = 0.019), weight-for-age Z-score, protein and fat intake, and time. In healthy infants, higher vitamin D status associates with leaner body composition, though the effect is smaller in magnitude relative to growth.
Size-independent organosilane functionalization of silicon nanocrystals using Wilkinson’s catalyst
Authors: Yang Zhenyu , Wahl Maike H. , Veinot Jonathan G.C.
Published: May 2014

A low-temperature size-independent method for modifying the surface chemistry of the silicon nanocrystal (SiNC) surface is reported. Wilkinson’s catalyst has been applied to accelerate the dehydrogenative coupling reaction between organosilane molecules and hydride-terminated SiNCs. Two strategies and multiple organosilanes were evaluated to study surface modification efficiency. During the investigations, it was determined that surface functionalization efficiency showed some dependence upon the organic modifier in question. The comparatively low reactivity of octadecyldimethyl silanes may result from the formation of “Si–Rh–Si” intermediates hindered by the steric bulk of the silanes when reacted with activated SiNCs. Quenching of the SiNC-based photoluminescence is observed and the origin of this phenomenon has been attributed to the present of trace rhodium on SiNCs detected using X-ray photoelectron spectroscopy.
Resonance Raman spectroscopic study of solvent-dependent coexistence of localized and delocalized dinitroaromatic radical anions
Authors: Hoekstra Ryan M. , Chen Yen-Ting , Kiesz Matthew D. , Telo João P. , Stephenson Rachel M. , Nelsen Stephen F. , Zink Jeffrey I.
Published: May 2014

Optical absorption spectra of three dinitroaromatic radical anions, 2,7-dinitro-9,9-dimethylfluorene (1•−), 4,4′-dinitrobiphenyl (2•−), and 4,4′-dinitrotolane (3•−) in solvents THF, HMPA, and MeCN show both an unresolved broad band characteristic of charge-localized mixed valence species (Robin-Day classification Class II), and a vibronically structured band of the delocalized species (Class III). With decreasing solvent reorganization energy, a greater portion of the compounds become charge-delocalized. In particular, 1•−with the greatest coupling, is almost entirely composed of the delocalized species in all solvents. An intense Raman mode is used to identify the charge-bearing unit as the nitrobenzene moiety. Resonance Raman profiles are utilized to gain detailed information of vibrational modes. The out-of-phase ring lengthening stretch mode is observed to be enhanced at higher excitation energies, corresponding to the absorption band of the Class II species, while the in-phase ring lengthening stretch mode, a totally symmetric vibration, is most strongly enhanced in the absorption region of the Class III species. Resonance Raman profiles support the solvent-dependent coexistence of Class II and Class III molecules of the same chemical composition.
Seismic performance of single-storey steel concentrically braced frame structures constructed in the 1960s
Authors: Caruso-Juliano A. , Gallagher A. , Morrison T.E. , Rogers C.A.
Published: May 2014

A general overview of the seismic performance of representative single-storey concentrically braced frame (CBF) buildings designed using the 1965 National Building Code of Canada and the 1965 S16 Canadian Standards Association Steel Structures for Buildings Standard was obtained through the use of nonlinear time history dynamic analyses and the testing of brace elements obtained from buildings constructed in the 1960s. The study comprised 16 representative buildings subjected to 20 site selected ground motions for three locations; Halifax (Nova Scotia) low seismicity, Montreal (Quebec) moderate seismicity, and Abbotsford (British Columbia) high seismicity. Incremental dynamic analyses were performed using various failure criteria as obtained from the test data; brace net-section fracture, bolt shear, block shear, bolt bearing and brace yielding. Fragility curves were created and the probability of failure was defined. The performance of the structures designed for Abbotsford was unsatisfactory for all of the brace connection failure mechanisms. In contrast, because of the lower current seismic hazard in Montreal, the 16 archetypical buildings demonstrated better performance. In Halifax the buildings performed well, based on the identified failure criteria. It was necessary to strengthen the roof diaphragms for all buildings to carry 2010 NBCC design level seismic forces.
Characterization of excavated claystone and claystone–bentonite mixtures as backfill/seal material
Authors: Zhang Chun-Liang
Published: May 2014

Crushed claystone produced by excavation of repository openings has been investigated as backfill/seal material. The raw coarse-grained claystone can be used for backfilling repository openings and, in mixture with bentonite, for sealing boreholes, drifts and shafts. The investigation programme focused on characterizing the thermo-hydro-mechanical properties of the excavated Callovo-Oxfordian claystone and the compacted claystone–bentonite mixtures, including (1) mechanical compaction, (2) gas and water permeability as a function of porosity, (3) water retention and saturation, (4) swelling capacity and (5) thermal properties of the materials. The major results are presented in this paper.
Altering the sex determination pathway inDrosophilafat body modifies sex-specific stress responses
Authors: Argue Kathryn J. , Neckameyer Wendi S.
Published: May 2014

The stress response in Drosophila melanogaster reveals sex differences in behavior, similar to what has been observed in mammals. However, unlike mammals, the sex determination pathway in Drosophila is well established, making this an ideal system to identify factors involved in the modulation of sex-specific responses to stress. In this study, we show that the Drosophila fat body, which has been shown to be important for energy homeostasis and sex determination, is a dynamic tissue that is altered in response to stress in a sex and time-dependent manner. We manipulated the sex determination pathway in the fat body via targeted expression of transformer and transformer-2 and analyzed these animals for changes in their response to stress. In the majority of cases, manipulation of transformer or transformer-2 was able to change the physiological output in response to starvation and oxidative stress to that of the opposite sex. Our data also uncover the possibility of additional downstream targets for transformer and transformer-2 that are separate from the sex determination pathway and can influence behavioral and physiological responses.
Role of prostaglandins in determining the increased cardiac sympathetic nerve activity in ovine sepsis
Authors: Booth Lindsea C. , Ramchandra Rohit , Calzavacca Paolo , May Clive N.
Published: May 2014

Effective treatment of sepsis remains a significant challenge in intensive care units. During sepsis, there is widespread activation of the sympathetic nervous system, which is thought to have both beneficial and detrimental effects. The sympathoexcitation is thought to be partly due to the developing hypotension, but may also be a response to the inflammatory mediators released. Thus, we investigated whether intracarotid infusion of prostaglandin E2 (PGE2) induced similar cardiovascular changes to those caused by intravenous infusion of Escherichia coli in sheep and whether inhibition of prostaglandin synthesis, with the nonselective cyclooxygenase inhibitor indomethacin, administered at 2 and 8 h after the onset of sepsis, reduced sympathetic nerve activity (SNA), and heart rate (HR). Studies were performed in conscious sheep instrumented to measure mean arterial pressure (MAP), HR, cardiac SNA (CSNA), and renal SNA (RSNA). Intracarotid infusion of PGE2 (50 ng·kg−1·min−1) increased temperature, CSNA, and HR, but not MAP or RSNA. Sepsis, induced by infusion of E. coli, increased CSNA, but caused an initial, transient inhibition of RSNA. At 2 h of sepsis, indomethacin (1.25 mg/kg bolus) increased MAP and caused reflex decreases in HR and CSNA. After 8 h of sepsis, indomethacin did not alter MAP, but reduced CSNA and HR, without altering baroreflex control. These findings indicate an important role for prostaglandins in mediating the increase in CSNA and HR during the development of hyperdynamic sepsis, whereas prostaglandins do not have a major role in determining the early changes in RSNA.