bluuebunny/crossref_metadata_2025 huggingface dataset:
Multisite Clinical Evaluation of a Rapid Test for Entamoeba histolytica in Stool
Authors: Verkerke Hans P. , Hanbury Blake , Siddique Abdullah , Samie Amidou , Haque Rashidul , Herbein Joel , Petri William A.
Published: November 2014

Rapid point-of-care detection of enteric protozoa in diarrheal stool is desirable in clinical and research settings to efficiently determine the etiology of diarrhea. We analyzed the ability of the third-generation
E. histolytica
Quik Chek assay developed by Techlab to detect amebic antigens in fecal samples collected from independent study populations in South Africa and Bangladesh. We compared the performance of this recently released rapid test to that of the commercially available ProSpecT
Entamoeba histolytica
microplate assay from Remel and the
E. histolytica
II enzyme-linked immunosorbent assay (ELISA) from Techlab, using real-time and nested-PCR for
Entamoeba
species to resolve any discrepant results. After discrepant resolution, The
E. histolytica
Quik Chek assay exhibited sensitivity and specificity compared to the
E. histolytica
II ELISA of 98.0% (95% confidence interval [CI], 92.9% to 99.8%) and 100% (95% CI, 99.0% to 100%), respectively. Compared to the ProSpecT microplate assay, the
E. histolytica
Quik Chek (Quik Chek) assay exhibited 97.0% sensitivity (95% CI, 91.5% to 99.4%) and 100% specificity (95% CI, 99.0% to 100%). Our results indicate that the Quik Chek is a robust assay for the specific detection of
E. histolytica
trophozoites in unfixed frozen clinical stool samples.
First Human Systemic Infection Caused by Spiroplasma
Authors: Aquilino Ana , Masiá Mar , López Pilar , Galiana Antonio J. , Tovar Juan , Andrés María , Gutiérrez Félix
Published: November 2014

Spiroplasma
species are organisms that normally colonize plants and insects. We describe the first case of human systemic infection caused by
Spiroplasma
bacteria in a patient with hypogammaglobulinemia undergoing treatment with biological disease-modifying antirheumatic agents.
Spiroplasma
turonicum
was identified through molecular methods in several blood cultures. The infection was successfully treated with doxycycline plus levofloxacin.
An Atypical Outbreak of Food-Borne Botulism Due to Clostridium botulinum Types B and E from Ham
Authors: Mazuet Christelle , Sautereau Jean , Legeay Christine , Bouchier Christiane , Bouvet Philippe , Popoff Michel R.
Published: November 2014

An outbreak of human botulism was due to consumption of ham containing botulinum neurotoxins B and E. A
Clostridium botulinum
type E strain isolated from ham was assigned to a new subtype (E12) based on
bont
/
E
gene sequencing and belongs to a new multilocus sequence subtype, as analyzed by whole-genome sequencing.
Gordonia Species as Emerging Causes of Continuous-Ambulatory-Peritoneal-Dialysis-Related Peritonitis Identified by 16S rRNA and
secA1
Gene Sequencing and Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS)
Authors: Lam Jimmy Y. W. , Wu Alan K. L. , Leung Wai-Shing , Cheung Ingrid , Tsang Chi-Ching , Chen Jonathan H. K. , Chan Jasper F. W. , Tse Cindy W. S. , Lee Rodney A. , Lau Susanna K. P. , Woo Patrick C. Y.
Published: November 2014

We report here four cases of continuous ambulatory peritoneal dialysis-related peritonitis caused by three different species of
Gordonia
. The portal of entry was likely through Tenckhoff catheters. 16S rRNA and s
ecA1
gene sequencing are so far the most reliable methods for the accurate identification of
Gordonia
species.
Trends in Secondary Antibiotic Resistance of Helicobacter pylori from 2007 to 2014: Has the Tide Turned?
Authors: Boltin Doron , Ben-Zvi Haim , Perets Tsachi Tsadok , Kamenetsky Zvi , Samra Zmira , Dickman Ram , Niv Yaron
Published: November 2014

The current guidelines recommend culture and antibiotic susceptibility testing of
Helicobacter pylori
following two failed eradication attempts. Where testing is unavailable, epidemiological data for secondary
H. pylori
resistance are essential to allow for the rational use of antibiotics. The aim of this study was to describe the temporal changes in antibiotic resistance among adults previously treated for
H. pylori
infections and to identify predictors of resistance. Between 2007 and 2014, consecutive patients undergoing gastroscopy with
H. pylori
culture and susceptibility testing at our institution following at least two treatment failures were retrospectively identified. Antibiotic susceptibilities were recorded and linked to the demographic data. A total of 1,042 patients were identified, including 739 (70.9%) males, aged 39.3 ± 18.9 years. Resistance to clarithromycin, metronidazole, and levofloxacin was found in 57.2%, 64.4%, and 5.1% of isolates, respectively. Dual resistance to clarithromycin and metronidazole was seen in 39.9%. Over the study period, clarithromycin resistance increased annually in a linear manner (odds ratio [OR], 1.09; 95% confidence interval [CI], 1.03 to 1.14;
P
< 0.01), levofloxacin resistance decreased annually (OR, 0.78; 95% CI, 0.61 to 0.92;
P
< 0.01), and metronidazole resistance was nonlinear. Age was an independent predictor of resistance to all antibiotics. Time elapsed predicted resistance for clarithromycin and levofloxacin and dual resistance for clarithromycin-metronidazole. Secondary resistance of
H. pylori
to clarithromycin and metronidazole remains high. The low secondary resistance to levofloxacin makes it an attractive treatment option in our region for patients following two failed eradication attempts.
Genomic Dissection of Travel-Associated Extended-Spectrum-Beta-Lactamase-Producing Salmonella enterica Serovar Typhi Isolates Originating from the Philippines: a One-Off Occurrence or a Threat to Effective Treatment of Typhoid Fever?
Authors: Hendriksen Rene S. , Leekitcharoenphon Pimlapas , Mikoleit Matthew , Jensen Jacob Dyring , Kaas Rolf Sommer , Roer Louise , Joshi Heena B. , Pornruangmong Srirat , Pulsrikarn Chaiwat , Gonzalez-Aviles Gladys D. , Reuland E. Ascelijn , Al Naiemi Nashwan , Wester Astrid Louise , Aarestrup Frank M. , Hasman Henrik
Published: November 2014

One unreported case of extended-spectrum-beta-lactamase (ESBL)-producing
Salmonella enterica
serovar Typhi was identified, whole-genome sequence typed, among other analyses, and compared to other available genomes of
S
. Typhi. The reported strain was similar to a previously published strain harboring
bla
SHV-12
from the Philippines and likely part of an undetected outbreak, the first of ESBL-producing
S
. Typhi.
Clinical Performance Evaluation of the Sofia RSV FIA Rapid Antigen Test for Diagnosis of Respiratory Syncytial Virus Infection
Authors: Jang Jin Woo , Cho Chi Hyun , Nam Myung-Hyun , Yoon Soo Young , Lee Chang Kyu , Lim Chae Seung , Kim Woo Joo
Published: November 2014

A recently introduced Sofia respiratory syncytial virus (RSV) fluorescent immunoassay (FIA) was evaluated against the BinaxNOW RSV card and the SD Bioline RSV test using 348 respiratory samples. The Sofia, BinaxNOW, and SD Bioline kits showed sensitivities of 66%, 65%, and 64%, respectively, for detecting RSV-A, and 71%, 63%, and 65% for detecting RSV-B, respectively.
Hydrophobic and Charged Residues in the C-Terminal Arm of Hepatitis C Virus RNA-Dependent RNA Polymerase Regulate Initiation and Elongation
Authors: Cherry Amy L. , Dennis Caitriona A. , Baron Andrew , Eisele Leslie E. , Thommes Pia A. , Jaeger Joachim
Published: November 2014

The RNA-dependent RNA polymerase (RdRp) of hepatitis C virus (HCV) is essential for viral genome replication. Crystal structures of the HCV RdRp reveal two C-terminal features, a β-loop and a C-terminal arm, suitably located for involvement in positioning components of the initiation complex. Here we show that these two elements intimately regulate template and nucleotide binding, initiation, and elongation. We constructed a series of β-loop and C-terminal arm mutants, which were used for
in vitro
analysis of RdRp
de novo
initiation and primer extension activities. All mutants showed a substantial decrease in initiation activities but a marked increase in primer extension activities, indicating an ability to form more stable elongation complexes with long primer-template RNAs. Structural studies of the mutants indicated that these enzyme properties might be attributed to an increased flexibility in the C-terminal features resulting in a more open polymerase cleft, which likely favors the elongation process but hampers the initiation steps. A UTP cocrystal structure of one mutant shows, in contrast to the wild-type protein, several alternate conformations of the substrate, confirming that even subtle changes in the C-terminal arm result in a more loosely organized active site and flexible binding modes of the nucleotide. We used a subgenomic replicon system to assess the effects of the same mutations on viral replication in cells. Even the subtlest mutations either severely impaired or completely abolished the ability of the replicon to replicate, further supporting the concept that the correct positioning of both the β-loop and C-terminal arm plays an essential role during initiation and in HCV replication in general.

IMPORTANCE
HCV RNA polymerase is a key target for the development of directly acting agents to cure HCV infections, which necessitates a thorough understanding of the functional roles of the various structural features of the RdRp. Here we show that even highly conservative changes, e.g., Tyr→Phe or Asp→Glu, in these seemingly peripheral structural features have profound effects on the initiation and elongation properties of the HCV polymerase.
Unraveling the Web of Viroinformatics: Computational Tools and Databases in Virus Research
Authors: Sharma Deepak , Priyadarshini Pragya , Vrati Sudhanshu
Published: November 2014

The beginning of the second century of research in the field of virology (the first virus was discovered in 1898) was marked by its amalgamation with bioinformatics, resulting in the birth of a new domain—viroinformatics. The availability of more than 100 Web servers and databases embracing all or specific viruses (for example, dengue virus, influenza virus, hepatitis virus, human immunodeficiency virus [HIV], hemorrhagic fever virus [HFV], human papillomavirus [HPV], West Nile virus, etc.) as well as distinct applications (comparative/diversity analysis, viral recombination, small interfering RNA [siRNA]/short hairpin RNA [shRNA]/microRNA [miRNA] studies, RNA folding, protein-protein interaction, structural analysis, and phylotyping and genotyping) will definitely aid the development of effective drugs and vaccines. However, information about their access and utility is not available at any single source or on any single platform. Therefore, a compendium of various computational tools and resources dedicated specifically to virology is presented in this article.
Nipah Virus Attachment Glycoprotein Stalk C-Terminal Region Links Receptor Binding to Fusion Triggering
Authors: Liu Qian , Bradel-Tretheway Birgit , Monreal Abrrey I. , Saludes Jonel P. , Lu Xiaonan , Nicola Anthony V. , Aguilar Hector C.
Published: November 2014

Membrane fusion is essential for paramyxovirus entry into target cells and for the cell-cell fusion (syncytia) that results from many paramyxoviral infections. The concerted efforts of two membrane-integral viral proteins, the attachment (HN, H, or G) and fusion (F) glycoproteins, mediate membrane fusion. The emergent Nipah virus (NiV) is a highly pathogenic and deadly zoonotic paramyxovirus. We recently reported that upon cell receptor ephrinB2 or ephrinB3 binding, at least two conformational changes occur in the NiV-G head, followed by one in the NiV-G stalk, that subsequently result in F triggering and F execution of membrane fusion. However, the domains and residues in NiV-G that trigger F and the specific events that link receptor binding to F triggering are unknown. In the present study, we identified a NiV-G stalk C-terminal region (amino acids 159 to 163) that is important for multiple G functions, including G tetramerization, conformational integrity, G-F interactions, receptor-induced conformational changes in G, and F triggering. On the basis of these results, we propose that this NiV-G region serves as an important structural and functional linker between the NiV-G head and the rest of the stalk and is critical in propagating the F-triggering signal via specific conformational changes that open a concealed F-triggering domain(s) in the G stalk. These findings broaden our understanding of the mechanism(s) of receptor-induced paramyxovirus F triggering during viral entry and cell-cell fusion.

IMPORTANCE
The emergent deadly viruses Nipah virus (NiV) and Hendra virus belong to the
Henipavirus
genus in the
Paramyxoviridae
family. NiV infections target endothelial cells and neurons and, in humans, result in 40 to 75% mortality rates. The broad tropism of the henipaviruses and the unavailability of therapeutics threaten the health of humans and livestock. Viral entry into host cells is the first step of henipavirus infections, which ultimately cause syncytium formation. After attaching to the host cell receptor, henipaviruses enter the target cell via direct viral-cell membrane fusion mediated by two membrane glycoproteins: the attachment protein (G) and the fusion protein (F). In this study, we identified and characterized a region in the NiV-G stalk C-terminal domain that links receptor binding to fusion triggering via several important glycoprotein functions. These findings advance our understanding of the membrane fusion-triggering mechanism(s) of the henipaviruses and the paramyxoviruses.
Lethal Coinfection of Influenza Virus and Streptococcus pneumoniae Lowers Antibody Response to Influenza Virus in Lung and Reduces Numbers of Germinal Center B Cells, T Follicular Helper Cells, and Plasma Cells in Mediastinal Lymph Node
Authors: Wu Yuet , Tu Wenwei , Lam Kwok-Tai , Chow Kin-Hung , Ho Pak-Leung , Guan Yi , Peiris Joseph S. Malik , Lau Yu-Lung
Published: November 2014

Secondary
Streptococcus pneumoniae
infection after influenza is a significant clinical complication resulting in morbidity and sometimes mortality. Prior influenza virus infection has been demonstrated to impair the macrophage and neutrophil response to the subsequent pneumococcal infection. In contrast, how a secondary pneumococcal infection after influenza can affect the adaptive immune response to the initial influenza virus infection is less well understood. Therefore, this study focuses on how secondary pneumococcal infection after influenza may impact the humoral immune response to the initial influenza virus infection in a lethal coinfection mouse model. Compared to mice infected with influenza virus alone, mice coinfected with influenza virus followed by pneumococcus had significant body weight loss and 100% mortality. In the lung, lethal coinfection significantly increased virus titers and bacterial cell counts and decreased the level of virus-specific IgG, IgM, and IgA, as well as the number of B cells, CD4 T cells, and plasma cells. Lethal coinfection significantly reduced the size and weight of spleen, as well as the number of B cells along the follicular developmental lineage. In mediastinal lymph nodes, lethal coinfection significantly decreased germinal center B cells, T follicular helper cells, and plasma cells. Adoptive transfer of influenza virus-specific immune serum to coinfected mice improved survival, suggesting the protective functions of anti-influenza virus antibodies. In conclusion, coinfection reduced the B cell response to influenza virus. This study helps us to understand the modulation of the B cell response to influenza virus during a lethal coinfection.

IMPORTANCE
Secondary pneumococcal infection after influenza virus infection is an important clinical issue that often results in excess mortality. Since antibodies are key mediators of protection, this study aims to examine the antibody response to influenza virus and demonstrates that lethal coinfection reduced the B cell response to influenza virus. This study helps to highlight the complexity of the modulation of the B cell response in the context of coinfection.
The nsp3 Macrodomain Promotes Virulence in Mice with Coronavirus-Induced Encephalitis
Authors: Fehr Anthony R. , Athmer Jeremiah , Channappanavar Rudragouda , Phillips Judith M. , Meyerholz David K. , Perlman Stanley
Published: November 2014

All coronaviruses encode a macrodomain containing ADP-ribose-1″-phosphatase (ADRP) activity within the N terminus of nonstructural protein 3 (nsp3). Previous work showed that mouse hepatitis virus strain A59 (MHV-A59) with a mutated catalytic site (N1348A) replicated similarly to wild-type virus but was unable to cause acute hepatitis in mice. To determine whether this attenuated phenotype is applicable to multiple disease models, we mutated the catalytic residue in the JHM strain of MHV (JHMV), which causes acute and chronic encephalomyelitis, using a newly developed bacterial artificial chromosome (BAC)-based MHV reverse genetics system. Infection of mice with the macrodomain catalytic point mutant virus (N1347A) resulted in reductions in lethality, weight loss, viral titers, proinflammatory cytokine and chemokine expression, and immune cell infiltration in the brain compared to mice infected with wild-type virus. Specifically, macrophages were most affected, with approximately 2.5-fold fewer macrophages at day 5 postinfection in N1347A-infected brains. Tumor necrosis factor (TNF) and interferon (IFN) signaling were not required for effective host control of mutant virus as all N1347A virus-infected mice survived the infection. However, the adaptive immune system was required for protection since N1347A virus was able to cause lethal encephalitis in RAG1
−/−
(recombination activation gene 1 knockout) mice although disease onset was modestly delayed. Overall, these results indicate that the BAC-based MHV reverse genetics system will be useful for studies of JHMV and expand upon previous studies, showing that the macrodomain is critical for the ability of coronaviruses to evade the immune system and promote viral pathogenesis.

IMPORTANCE
Coronaviruses are an important cause of human and veterinary diseases worldwide. Viral processes that are conserved across a family are likely to be good targets for the development of antiviral therapeutics and vaccines. The macrodomain is a ubiquitous structural domain and is also conserved among all coronaviruses. The coronavirus macrodomain has ADP-ribose-1″-phosphatase activity; however, its function during infection remains unclear as does the reason that coronaviruses have maintained this enzymatic activity throughout evolution. For MHV, this domain has now been shown to promote multiple types of disease, including hepatitis and encephalitis. These data indicate that this domain is vital for the virus to replicate and cause disease. Understanding the mechanism used by this enzyme to promote viral pathogenesis will open up novel avenues for therapies and may give further insight into the role of macrodomain proteins in the host cell since these proteins are found in all living organisms.
Interferon-Induced Ifit Proteins: Their Role in Viral Pathogenesis
Authors: Fensterl Volker , Sen Ganes C.
Published: November 2014

A major component of the protective antiviral host defense is contributed by the intracellular actions of the proteins encoded by interferon-stimulated genes (ISGs); among these are the
i
nter
f
eron-
i
nduced proteins with
t
etratricopeptide repeats (IFITs), consisting of four members in human and three in mouse. IFIT proteins do not have any known enzyme activity. Instead, they inhibit virus replication by binding and regulating the functions of cellular and viral proteins and RNAs. Although all IFITs are comprised of multiple copies of the degenerate tetratricopeptide repeats, their distinct tertiary structures enable them to bind different partners and affect host-virus interactions differently. The recent use of
Ifit
knockout mouse models has revealed novel antiviral functions of these proteins and new insights into the specificities of ISG actions. This article focuses on human and murine IFIT1 and IFIT2 by reviewing their mechanisms of action, their critical roles in protecting mice from viral pathogenesis, and viral strategies to evade IFIT action.
Human Noroviruses' Fondness for Histo-Blood Group Antigens
Authors: Singh Bishal K. , Leuthold Mila M. , Hansman Grant S.
Published: November 2014

Human noroviruses are the dominant cause of outbreaks of gastroenteritis around the world. Human noroviruses interact with the polymorphic human histo-blood group antigens (HBGAs), and this interaction is thought to be important for infection. Indeed, synthetic HBGAs or HBGA-expressing enteric bacteria were shown to enhance norovirus infection in B cells. A number of studies have found a possible relationship between HBGA type and norovirus susceptibility. The genogroup II, genotype 4 (GII.4) noroviruses are the dominant cluster, evolve every other year, and are thought to modify their binding interactions with different HBGA types. Here we show high-resolution X-ray crystal structures of the capsid protruding (P) domains from epidemic GII.4 variants from 2004, 2006, and 2012, cocrystallized with a panel of HBGA types (H type 2, Lewis Y, Lewis B, Lewis A, Lewis X, A type, and B type). Many of the HBGA binding interactions were found to be complex, involving capsid loop movements, alternative HBGA conformations, and HBGA rotations. We showed that a loop (residues 391 to 395) was elegantly repositioned to allow for Lewis Y binding. This loop was also slightly shifted to provide direct hydrogen- and water-mediated bonds with Lewis B. We considered that the flexible loop modulated Lewis HBGA binding. The GII.4 noroviruses have dominated outbreaks over the past decade, which may be explained by their exquisite HBGA binding mechanisms, their fondness for Lewis HBGAs, and their temporal amino acid modifications.

IMPORTANCE
Our data provide a comprehensive picture of GII.4 P domain and HBGA binding interactions. The exceptionally high resolutions of our X-ray crystal structures allowed us to accurately recognize novel GII.4 P domain interactions with numerous HBGA types. We showed that the GII.4 P domain-HBGA interactions involved complex binding mechanisms that were not previously observed in norovirus structural studies. Many of the GII.4 P domain-HBGA interactions we identified were negative in earlier enzyme-linked immunosorbent assay (ELISA)-based studies. Altogether, our data show that the GII.4 norovirus P domains can accommodate numerous HBGA types.
Characterization of a Proteolytically Stable D-Peptide That Suppresses Herpes Simplex Virus 1 Infection: Implications for the Development of Entry-Based Antiviral Therapy
Authors: Jaishankar Dinesh , Yakoub Abraam M. , Bogdanov Anita , Valyi-Nagy Tibor , Shukla Deepak
Published: November 2014

Uncontrolled herpes simplex virus 1 (HSV-1) infection can advance to serious conditions, including corneal blindness or fatal encephalitis. Here, we describe a highly potent anti-HSV-1 peptide (DG2) that inhibits HSV-1 entry into host cells and blocks all aspects of infection. Importantly, DG2 is highly resistant to proteases and shows minimal toxicity, paving the way for prophylactic or therapeutic application of the peptide
in vivo
.
The Enterovirus 71 Procapsid Binds Neutralizing Antibodies and Rescues Virus Infection
In Vitro
Authors: Shingler Kristin L. , Cifuente Javier O. , Ashley Robert E. , Makhov Alexander M. , Conway James F. , Hafenstein Susan
Published: November 2014

Enterovirus 71 (EV71) is responsible for seasonal outbreaks of hand, foot, and mouth disease in the Asia-Pacific region. The virus has the capability to cause severe disease and death, especially in young children. Although several vaccines are currently in clinical trials, no vaccines or therapeutics have been approved for use. Previous structural studies have revealed that two antigenically distinct capsid forms are produced in EV71-infected cells: an expanded empty capsid, sometimes called a procapsid, and the infectious virus. Specifically, an immunodominant epitope of EV71 that maps to the virus canyon is structurally different in the procapsid and virus. This structure-function study shows that the procapsid can sequester antibodies, thus enhancing EV71 infection
in vitro
. The results presented here suggest that, due to conformational differences between the EV71 procapsid and virus, the presence of the procapsid in natural virus infections should be considered in the future design of vaccines or therapeutics.

IMPORTANCE
In a picornavirus infection, both an infectious and a noninfectious empty capsid, sometimes referred to as a procapsid, are produced. It was novel to discover that the procapsid form of EV71 was expanded and antigenically distinct from the infectious virus. Previously, it had been supposed that this empty capsid was an off-pathway dead end or at best served for storage of pentameric subunits, which was later shown to be unlikely. It remains unexplained why picornaviruses evolutionarily conserve the wasteful production of so much noninfectious capsid. Here, we demonstrate that the EV71 procapsid has different antigenic properties than the infectious virus. Thus, the procapsid has the capacity to sequester neutralizing antibody and protect the virus, promoting or restoring a successful infection
in vitro
. This important observation should be considered in the future design and development of vaccines and therapeutics.
Influenza A Virus Uses Intercellular Connections To Spread to Neighboring Cells
Authors: Roberts Kari L. , Manicassamy Balaji , Lamb Robert A.
Published: November 2014

In the extracellular environment, cell-free virions seek out naive host cells over long distances and between organisms. This is the primary mechanism of spread for most viruses. Here we provide evidence for an alternative pathway previously undescribed for orthomyxoviruses, whereby the spread of influenza A virus (IAV) infectious cores to neighboring cells can occur within intercellular connections. The formation of these connections requires actin dynamics and is enhanced by viral infection. Connected cells have contiguous membranes, and the core infectious viral machinery (RNP and polymerase) was present inside the intercellular connections. A live-cell movie of green fluorescent protein (GFP)-tagged NS1 of IAV shows viral protein moving from one cell to another through an intercellular connection. The movement of tagged protein was saltatory but overall traveled only in one direction. Infectious virus cores can move from one cell to another without budding and release of cell-free virions, as evidenced by the finding that whereas a neuraminidase inhibitor alone did not inhibit the development of IAV microplaques, the presence of a neuraminidase inhibitor together with drugs inhibiting actin dynamics or the microtubule stabilizer paclitaxel (originally named taxol) precluded microplaque formation. Similar results were also observed with parainfluenza virus 5 (PIV5), a paramyxovirus, when neutralizing antibody was used to block spread by cell-free virions. Intercellular spread of infectious core particles was unaffected or enhanced in the presence of nocodazole for IAV but inhibited for PIV5. The intercellular connections have a core of filamentous actin, which hints toward transport of virus particles through the use of a myosin motor.

IMPORTANCE
Here we describe a new method by which influenza A virus (IAV) spreads from cell to cell: IAV uses intracellular connections. The formation of these connections requires actin dynamics and is enhanced by viral infection and the absence of microtubules. Connected cells appeared to have contiguous membranes, and the core infectious viral machinery (RNP and polymerase) was present inside the intercellular connections. Infectious virus cores can move from one cell to another without budding and release of cell-free virions. Similar results were also observed with parainfluenza virus 5 (PIV5).
Identification of a Role for Nucleolin in Rabies Virus Infection
Authors: Oksayan S. , Nikolic J. , David C. T. , Blondel D. , Jans D. A. , Moseley G. W.
Published: November 2014

Rabies virus replicates in the cytoplasm of host cells, but rabies virus phosphoprotein (P-protein) undergoes active nucleocytoplasmic trafficking. Here we show that the largely nuclear P-protein isoform P3 can localize to nucleoli and forms specific interactions with nucleolin. Importantly, depletion of nucleolin expression inhibits viral protein expression and infectious virus production by infected cells. This provides the first evidence that lyssaviruses interact with nucleolin and that nucleolin is important to lyssavirus infection.
Paracetamol exposure in pregnancy and early childhood and development of childhood asthma: a systematic review and meta-analysis
Authors: Cheelo M , Lodge C J , Dharmage S C , Simpson J A , Matheson M , Heinrich J , Lowe A J
Published: November 2014

Background and objective

While paracetamol exposure in pregnancy and early infancy has been associated with asthma, it remains unclear whether this is confounded by respiratory tract infections, which have been suggested as an alternative explanation. We undertook a systematic review and meta-analysis of longitudinal studies that reported the association between paracetamol exposure during pregnancy or infancy and the subsequent development of childhood asthma (≥5 years).

Methods

Two independent researchers searched the databases EMBASE and PUBMED on 12 August 2013 for relevant articles using predefined inclusion and exclusion criteria. Study quality was assessed and results were pooled using fixed effect models or random effect models when moderate between-study heterogeneity was observed. We explicitly assessed whether the observed associations are due to confounding by respiratory tract infections.

Results

Eleven observational cohort studies met the inclusion criteria. Any paracetamol use during the first trimester was related to increased risk of childhood asthma (5 studies, pooled OR=1.39, 95% CI 1.01 to 1.91) but there was marked between-study heterogeneity (I2=63%) and only one of these studies adjusted for maternal respiratory tract infections. Increasing frequency of use of paracetamol during infancy was associated with increased odds of childhood asthma (3 studies, pooled OR=1.15, 95% CI 1.00 to 1.31 per doubling of days exposure), but in these same three studies adjusting for respiratory tract infections reduced this association (OR=1.06, 95% CI 0.92 to 1.22).

Discussion

The association during early pregnancy exposure was highly variable between studies and exposure during infancy appears to be moderately confounded by respiratory tract infections. There is insufficient evidence to warrant changing guidelines on early life paracetamol exposure at this time.
Longitudinal follow-up of employment status in patients with chronic fatigue syndrome after mononucleosis
Authors: Nyland Morten , Naess Halvor , Birkeland Jon Steinar , Nyland Harald
Published: November 2014

Objective

To examine the effect of early clinical and demographic factors on occupational outcome, return to work or awarded permanent disability pension in young patients with chronic fatigue syndrome (CFS).

Design

Longitudinal cohort study.

Intervention

A written self-management programme including a description of active coping strategies for daily life was provided.

Setting, participants

Patients with CFS after mononucleosis were evaluated at Department of Neurology, Haukeland University Hospital during 1996–2006 (contact 1). In 2009 self-report questionnaires were sent to all patients (contact 2).

Primary and secondary outcome measures

Primary measure was employment status at contact 2.Secondary measures included clinical symptoms, and Fatigue Severity Scale (FSS) scores on both contacts, and Work and Social Adjustment Scale (WSAS) at contact 2.

Results

Of 111 patients at contact 1, 92 (83%) patients returned the questionnaire at contact 2. Mean disease duration at contact 1 was 4.7 years and at contact 2 11.4 years. At contact 1, 9 (10%) were part-time or full-time employed. At contact 2, 49 (55%) were part-time or full-time employed. Logical regression analysis showed that FSS≥5 at contact 2 was associated with depression, arthralgia and long disease duration (all at contact 1).

Conclusions

About half of younger patients with CFS with long-term incapacity for work experienced marked improvement including full-time or part-time employment showing better outcomes than expected. Risk factors for transition to permanent disability were depression, arthralgia and disease duration.
The population-based Occupational and Environmental Health Prospective Cohort Study (AMIGO) in the Netherlands
Authors: Slottje Pauline , Yzermans C Joris , Korevaar Joke C , Hooiveld Mariëtte , Vermeulen Roel C H
Published: November 2014

Purpose

Occupational and environmental exposures remain important modifiable risk factors of public health. Existing cohort studies are often limited by the level of detail of data collected on these factors and health. It is also often assumed that the more healthy group is over-represented in cohort studies, which is of concern for their external validity. In this cohort profile, we describe how we set up the population-based Occupational and Environmental Health Cohort Study (AMIGO) to longitudinally study occupational and environmental determinants of diseases and well-being from a multidisciplinary and life course point of view. Reviewed by the Medical Ethics Research Committee of the University Medical Center Utrecht (protocol 10-268/C). All cohort members participate voluntarily and gave informed consent prior to their inclusion.

Participants

14 829 adult cohort members (16% of those invited) consented and filled in the online baseline questionnaire. Determinants include chemical, biological, physical (eg, electromagnetic fields), and psychosocial factors. Priority health outcomes include cancer, neurological, cardiovascular and respiratory diseases and non-specific symptoms. Owing to the recruitment strategy via general practitioners of an established network, we also collect longitudinal data registered in their electronic medical records including symptoms, diagnosis and treatments. Besides the advantage of health outcomes that cannot be easily captured longitudinally by other means, this created a unique opportunity to assess health-related participation bias by comparing general practitioner-registered prevalence rates in the cohort and its source population.

Findings to date

We found no indications of such a systematic bias. The major assets of the AMIGO approach are its detailed occupational and environmental determinants in combination with the longitudinal health data registered in general practice besides linkage to cancer and mortality registries and self-reported health.

Future plans

We are now in the phase of prospective follow-up, with the aim of continuing this for as long as possible (20+ years), pending future funding. Findings will be disseminated through scientific conferences and peer-reviewed journals, and through newsletters and the project website to participants, stakeholders and the wider public.
Emerging role of traditional birth attendants in mountainous terrain: a qualitative exploratory study from Chitral District, Pakistan
Authors: Shaikh Babar Tasneem , Khan Sharifullah , Maab Ayesha , Amjad Sohail
Published: November 2014

Objectives

This research endeavours to identify the role of traditional birth attendants (TBAs) in supporting the maternal, newborn and child health (MNCH) care, partnership mechanism with a formal health system and also explored livelihood options for TBAs in the health system of Pakistan.

Setting

The study was conducted in district Chitral, Khyber Pakhtunkhwa province, covering the areas where the Chitral Child Survival programme was implemented.

Participants

A qualitative exploratory study was conducted, comprising seven key informant interviews with health managers, and four focus group discussions with community midwives (CMWs), TBAs, members of Community Based Saving Groups (CBSGs) and members of village health committees (VHCs).

Results

The study identified that in the new scenario, after the introduction of CMWs in the health system, TBAs still have a pivotal role in health promotion activities such as breastfeeding promotion and vaccination. TBAs can assist CMWs in normal deliveries, and refer high-risk cases to the formal health system. Generally, TBAs are positive about CMWs’ introduction and welcome this addition. Yet their livelihood has suffered after CMWs’ deployment. Monetary incentives to them in recognition of referrals to CMWs could be one solution. The VHC is an active forum for strengthening co-ordination between the two service providers and to ensure an alternate and permanent livelihood support system for the TBAs.

Conclusions

TBAs have assured their continued support in provision of continuum of care for pregnant women, lactating mothers and children under the age of 5 years. The district health authorities must figure out ways to foster a healthy interface vis-à-vis roles and responsibilities of TBAs and CMWs. In time it would be worthwhile to do further research to look into the CMWs’ integration in the system, as well as TBAs’ continued role for provision of MNCH care.
Evaluating the efficacy of thoracoscopy and talc poudrage versus pleurodesis using talc slurry (TAPPS trial): protocol of an open-label randomised controlled trial
Authors: Bhatnagar Rahul , Laskawiec-Szkonter Magda , Piotrowska Hania E G , Kahan Brennan C , Hooper Clare E , Davies Helen E , Harvey John E , Miller Robert F , Rahman Najib M , Maskell Nick A
Published: November 2014

Introduction

The management of recurrent malignant pleural effusions (MPE) can be challenging. Various options are available, with the most efficacious and widely used being talc pleurodesis. Talc can either be applied via a chest drain in the form of slurry, or at medical thoracoscopy using poudrage. Current evidence regarding which method is most effective is conflicting and often methodologically flawed. The TAPPS trial is a suitably powered, multicentre, open-label, randomised controlled trial designed to compare the pleurodesis success rate of medical thoracoscopy and talc poudrage with chest drain insertion and talc slurry.

Methods and analysis

330 patients with a confirmed MPE requiring intervention will be recruited from UK hospitals. Patients will be randomised (1:1) to undergo either small bore (<14 Fr) Seldinger chest drain insertion followed by instillation of sterile talc (4 g), or to undergo medical thoracoscopy and simultaneous poudrage (4 g). The allocated procedure will be performed as an inpatient within 3 days of randomisation taking place. Following discharge, patients will be followed up at regular intervals for 6 months. The primary outcome measure is pleurodesis failure rates at 3 months. Pleurodesis failure is defined as the need for further pleural intervention for fluid management on the side of the trial intervention.

Ethics and dissemination

The trial has received ethical approval from the National Research Ethics Service Committee North West—Preston (12/NW/0467). There is a trial steering committee which includes independent members and a patient and public representative. The trial results will be published in a peer-reviewed journal and presented at international conferences, as well as being disseminated via local and national charities and patient groups. All participants who wish to know the study results will also be contacted directly on their publication.

Trial registration number

ISRCTN47845793.
Internet use, social engagement and health literacy decline during ageing in a longitudinal cohort of older English adults
Authors: Kobayashi Lindsay C , Wardle Jane , von Wagner Christian
Published: November 2014

Background

Health literacy skills tend to decline during ageing, which is often attributed to age-related cognitive decline. Whether health literacy skills may be influenced by technological and social factors during ageing is unknown.

Methods

We investigated whether internet use and social engagement protect against health literacy decline during ageing, independent of cognitive decline. We used prospective data from 4368 men and women aged ≥52 years in the English Longitudinal Study of Ageing from 2004 to 2011. Health literacy was measured at baseline (2004–2005) and at follow-up (2010–2011) using a reading comprehension test of a fictitious medicine label. The influences of consistent internet use and engagement in each of the civic, leisure and cultural activities on health literacy decline over the follow-up were estimated.

Results

After adjusting for cognitive decline and other covariates, consistent internet use (1379/4368; 32%) was protectively associated with health literacy decline (OR=0.77; 95% CI 0.60 to 0.99), as was consistent engagement in cultural activities (1715/4368; 39%; OR=0.73; 95% CI 0.56 to 0.93). As the number of activities engaged in increased, the likelihood of health literacy decline steadily decreased (ptrend<0.0001), with OR=0.51 (95% CI 0.33 to 0.79) for engaging in all four of the internet use and civic, leisure and cultural activities versus none.

Conclusions

Internet use and social engagement, particularly in cultural activities (eg, attending the cinema, art galleries, museums and the theatre), may help older adults to maintain health literacy during ageing. Support for older adults to maintain socially engaged lives and to access the internet should help promote the maintenance of functional literacy skills during ageing.
Influence of running shoes and cross-trainers on Achilles tendon forces during running compared with military boots
Authors: Sinclair Jonathan , Taylor P J , Atkins S
Published: November 2014

Military recruits are known to be susceptible to Achilles tendon pathology. The British Army have introduced footwear models, the PT-03 (cross-trainer) and PT1000 (running shoes), in an attempt to reduce the incidence of injuries. The aim of the current investigation was to examine the Achilles tendon forces of the cross-trainer and running shoe in relation to conventional army boots. Ten male participants ran at 4.0 m/s in each footwear condition. Achilles tendon forces were obtained throughout the stance phase of running and compared using repeated-measures ANOVAs. The results showed that the time to peak Achilles tendon force was significantly shorter when running in conventional army boots (0.12 s) in comparison with the cross-trainer (0.13 s) and running shoe (0.13 s). Achilles tendon loading rate was shown to be significantly greater in conventional army boots (38.73 BW/s) in comparison with the cross-trainer (35.14 BW/s) and running shoe (33.57 BW/s). The results of this study suggest that the running shoes and cross-trainer footwear are associated with reductions in Achilles tendon parameters that have been linked to the aetiology of injury, and thus it can be hypothesised that these footwear could be beneficial for military recruits undertaking running exercises.
Republished: Bilateral carotid and vertebral rete mirabile with vein of Galen aneurysmal malformation: an unreported association
Authors: Mondel Prabath Kumar , Saraf Rashmi , Limaye Uday S
Published: November 2014

Rete mirabile is a fine meshwork of anastomosing vessels that replace the parent artery. A 30-year-old woman complained of slurring of speech, right eye proptosis, recurrent vomiting, and loss of bladder and bowel control, followed by drowsiness lasting 30–40 min, for the past 6 months. On cross sectional imaging and angiography, the patient was found to have a vein of Galen aneurysmal malformation, with bilateral carotid and vertebral rete mirabile. The patient was offered both endovascular and open surgical options but she refused any form of surgical treatment and opted for conservative management. At the 6 month follow-up, she continued to have occasional episodes of headache and vomiting but was otherwise normal. We describe the clinical, cross sectional, and angiographic features of this patient. A comparison with other patients with bilateral carotid and vertebral rete mirabile is also reported.
Assisted coiling of saccular wide-necked unruptured intracranial aneurysms: stent versus balloon
Authors: Consoli Arturo , Vignoli Chiara , Renieri Leonardo , Rosi Andrea , Chiarotti Ivano , Nappini Sergio , Limbucci Nicola , Mangiafico Salvatore
Published: November 2014

Background and purpose

Assisted coiling with stents or balloons enables a higher percentage of complete occlusions of saccular unruptured intracranial aneurysms to be achieved with a reasonable complication rate. The aim of this study was to compare stent-assisted coiling and the balloon remodeling technique in terms of efficacy, stability, and safety for the treatment of comparable unruptured saccular intracranial aneurysms.

Materials and methods

268 patients with 286 saccular unruptured wide-necked intracranial aneurysms were treated at our institution with stent- or balloon-assisted coiling and retrospectively reviewed. Statistical analysis was performed to assess significant differences between the two groups.

Results

The rate of complete occlusion at the end of the procedure was higher with stent-assisted coiling than with balloon-assisted coiling (86.8% vs 78%) and the same results were also observed after 6 months (92.1% vs 77.6%; p=0.05). About 50% of major recurrences occurred in large to giant aneurysms (p<0.001). The overall complication rate was similar in the stent-assisted and balloon-assisted groups (10.3% vs 9.3%). Independently of the technique, a higher complication rate was observed with bifurcational aneurysms, particularly in the middle cerebral artery (p=0.016).

Conclusions

Stent-assisted coiling achieved better results in terms of complete occlusion and stability than balloon-assisted coiling with a lower rate of recurrence without being associated with a higher risk of intraprocedural complications. Bifurcational and large to giant aneurysms were associated with higher complication rates and higher recurrence rates, respectively, and still represent a challenge for both techniques.
Remission of neurovascular conflicts in the cerebellopontine angle in interventional neuroradiology
Authors: Li Chuanhui , Li Youxiang , Jiang Chuhan , Wu Zhongxue , Wang Yang , Yang Xinjian
Published: November 2014

Background and purpose

To investigate the efficacy of endovascular treatment (EVT) for neurovascular conflicts (NVCs) in the cerebellopontine angle (CPA) caused by intracranial aneurysms (IAs) and intracranial arteriovenous malformations (AVMs), including trigeminal neuralgia, hemifacial spasm, and glossopharyngeal neuralgia.

Materials and methods

From January 2010 to January 2014, 14 consecutive patients presenting with three NVCs caused by IAs or intracranial AVMs were admitted to our department. The clinical outcomes of these NVCs after EVT were retrospectively analyzed.

Results

For four patients with IAs, angiographic follow-up confirmed total occlusion of the lesion in all, and the clinical outcomes of NVC were as follows: gradual relief in two (50%), transient partial relief but recurrence in one (25%), and no palliative effect in one (25%). For the 10 patients with intracranial AVMs, one (10%) experienced transient relief of NVC after angiogram examination (no EVT was performed). Of the other nine patients who received EVT, angiographic follow-up was obtained in seven (70%), demonstrating total obliteration of the lesion in three (30%), subtotal obliteration in two (20%), and partial obliteration in two (20%). Clinical outcomes included immediate relief of NVCs after single EVT in two cases (20%), gradual relief after single EVT in five (50%, one of them experienced transient aggravation), and complete relief after two sessions of EVT in two (20%). Complications of transient cranial nerve paresis related to EVT occurred in two cases (20%) with intracranial AVMs. In all, complete lasting relief of the NVCs was obtained finally in 11 cases (78.6%).

Conclusions

EVT is a feasible and less invasive approach for relief of NVCs in the CPA caused by IA or intracranial AVM and could be considered as a therapeutic option in these situations.