bluuebunny/crossref_metadata_2025 huggingface dataset:
Silencing of the gibberellin receptor homolog, CsGID1a, affects locule formation in cucumber (Cucumis sativus) fruit
Authors: Liu Bin , Liu Xingwang , Yang Sen , Chen Chunhua , Xue Shudan , Cai Yanling , Wang Dandan , Yin Shuai , Gai Xinshuang , Ren Huazhong
Published: December 2015

Gibberellins are phytohormones with many roles, including the regulation of fruit development. However, little is known about the relationship between GA perception and fleshy fruit ontogeny, and particularly locule formation.

We characterized the expression of cucumber (Cucumis sativus) GA receptor gene (CsGID1a) using quantitative real‐time PCR, in situ hybridization and a promoter::β‐glucuronidase (GUS) assay. CsGID1a‐RNAi cucumber fruits were observed by dissecting microscope, scanning electron microscopy and transmission electron microscopy. Finally, genome‐wide gene expression in young fruits from a control and the RNAi line was compared using a digital gene expression (DGE) analysis approach.

The expression pattern of CsGID1a was found to be closely correlated with fruit locule formation, and silencing CsGID1a in cucumber resulted in fruits with abnormal carpels and locules. Overexpression of CsGID1a in the Arabidopsis thaliana double mutant (gid1a gid1c) resulted in ‘cucumber locule‐like’ fruits. The DGE analysis suggested that expression of genes related to auxin synthesis and transport, as well as the cell cycle, was altered in CsGID1a‐RNAi fruits, a result that was supported by comparing the auxin content and cellular structures of the control and transgenic fruits.

This study demonstrates a previously uncharacterized GA signaling pathway that is essential for cucumber fruit locule formation.
Autonomic and sensory ganglionopathy occurring in a patient with fulminant type 1 diabetes mellitus
Authors: Makino Mutsuki , Hiwatashi Dai , Minemura Kesami , Kawaguchi Kenji
Published: December 2015

A 63‐year‐old male with a previous infection was admitted to our hospital because of acute pancreatitis. Although he had no history of diabetes mellitus, laboratory examinations revealed marked hyperglycemia on admission, and intensive insulin treatment was required. After 2 weeks, he developed severe pandysautonomia and sensory impairment, and eventually died from colonic perforation caused by paralytic ileus at 1 year after onset. Autopsy findings showed a complete loss of pancreatic islet beta cells with mild fibrosis of the exocrine pancreas. Neuropathological examination showed myelin loss of the dorsal fasciculus at all levels of the spinal cord and ganglionopathy of the dorsal root and sympathetic truncus, revealing degeneration of ganglion cells and a decrease in their number with the formation of Nageotte nodules. On the basis of this clinicopathological evidence, fulminant type 1 diabetes mellitus (T1DM) and idiopathic immune‐mediated autonomic neuropathy were suspected. Here, we suggest a close etiological relationship between both diseases.
Foetal and infant growth patterns, airway resistance and school‐age asthma
Authors: Sonnenschein‐van der Voort Agnes M.M. , Gaillard Romy , de Jongste Johan C. , Hofman Albert , Jaddoe Vincent W.V. , Duijts Liesbeth
Published: December 2015

Background and objective

Preterm birth, low birth weight and rapid infant weight gain are associated with increased risks of asthma symptoms in childhood. The underlying mechanism may include persistently higher airway resistance (Rint). The aim of our study was to examine the associations of longitudinally measured foetal and infant growth characteristics with Rint and asthma outcomes in school‐age children.

Methods

This study was embedded in a population‐based prospective cohort study in Rotterdam, The Netherlands. Foetal growth was estimated by ultrasound in the second and third trimesters. Infant growth was measured at birth, 3, 6 and 12 months. At age 6 years, Rint was measured, and information about wheezing and asthma was obtained by questionnaires. The number of subjects per analysis differed per available outcome (3954–5066 subjects).

Results

Longitudinal growth analyses showed that school‐age children with increased Rint had lower foetal length growth and weight gain, and lower infant length growth. Children with persistent wheezing until age 6 years and physician‐diagnosed asthma had a higher Rint compared with children who never wheezed or without asthma (difference z‐scores Rint: 0.58 (0.19, 0.97) and 0.55 (0.15, 0.95), respectively).

Conclusion

Rint in school‐age children is influenced by foetal growth restriction and is associated with asthma outcomes.

See Editorial, page 574
You Catch More Flies With Honey: Sex Work, Violence, and Masculinity on the Streets
Authors: Oselin Sharon S.
Published: December 2015

Violence among inner‐city men is a pressing social concern, and the central focus of much academic research. Many frame it as a phenomenon that certain men perpetuate—those who inhabit disadvantaged, impoverished communities—and argue it is linked to performances of “street” masculinity. In this article, I examine male street‐based sex workers’ willingness to become embroiled in violent exchanges. In a departure from theoretical predictions, my findings reveal these men expend considerable effort to remain nonviolent with others immersed in the sex trade, a decision based upon their desire for the acquisition of capital as well as their calculation of risks. In doing so, they construct and perform a nuanced version of masculinity, which I call pacifist masculinity. Few studies analyze peaceful and conciliatory interactions among men in these contexts, an absence that only serves to reify assumptions about rampant hostility and aggression. I draw on interviews with 19 men involved in street prostitution in Chicago in 2012. This article contributes to a clearer understanding of male–male violence in high‐risk environments, examines the prominent factors that inform decisions to assault others, and explores how such actions challenge hegemonic masculinity.
Seroprevalence of HTLV‐1 and HTLV‐2 amongst mothers and children in Malawi within the context of a systematic review and meta‐analysis of HTLV seroprevalence in Africa
Authors: Fox James M. , Mutalima Nora , Molyneux Elizabeth , Carpenter Lucy M. , Taylor Graham P. , Bland Martin , Newton Robert , Martin Fabiola
Published: December 2015

Objectives

Human T‐lymphotropic virus (HTLV)‐1 causes T‐cell leukaemia and myelopathy. Together with HTLV‐2, it is endemic in some African nations. Seroprevalence data from Malawi are scarce, with no reports on associated disease incidence. HTLV seroprevalence and type were tested in 418 healthy mothers from Malawi. In addition, we tested the sera of 534 children to investigate mother‐to‐child transmission. To provide context, we conducted a systematic review and meta‐analysis of HTLV seroprevalence in African women and children.

Methods

Stored samples from a previous childhood cancer and BBV study were analysed. ELISA was used for HTLV screening followed by immunoblot for confirmation and typing. Standard methods were used for the systematic review.

Results

HTLV seroprevalence was 2.6% (11/418) in mothers and 2.2% (12/534) in children. Three mothers carried HTLV‐1 alone, seven had HTLV‐2 and one was dually infected. Three children carried HTLV‐1 alone, seven had HTLV‐2 and two were dually infected. Only two corresponding mothers of the 12 HTLV‐positive children were HTLV positive. The systematic review included 66 studies of women and 13 of children conducted in 25 African countries. Seroprevalence of HTLV‐1 varied from 0 to 17% and of HTLV‐2 from 0 to 4%.

Conclusions

In contrast to findings from other studies in Africa, the seroprevalence of HTLV‐2 was higher than that of HTLV‐1 in Malawi and one of the highest for the African region. The lack of mother–child concordance suggests alternative sources of infection among children. Our data and analyses contribute to HTLV prevalence mapping in Africa.
Spatial atomic layer deposition on flexible porous substrates: ZnO on anodic aluminum oxide films and Al2O3 on Li ion battery electrodes
Authors: Sharma Kashish , Routkevitch Dmitri , Varaksa Natalia , George Steven M.
Published: December 2015

Spatial atomic layer deposition (S-ALD) was examined on flexible porous substrates utilizing a rotating cylinder reactor to perform the S-ALD. S-ALD was first explored on flexible polyethylene terephthalate polymer substrates to obtain S-ALD growth rates on flat surfaces. ZnO ALD with diethylzinc and ozone as the reactants at 50 °C was the model S-ALD system. ZnO S-ALD was then performed on nanoporous flexible anodic aluminum oxide (AAO) films. ZnO S-ALD in porous substrates depends on the pore diameter, pore aspect ratio, and reactant exposure time that define the gas transport. To evaluate these parameters, the Zn coverage profiles in the pores of the AAO films were measured using energy dispersive spectroscopy (EDS). EDS measurements were conducted for different reaction conditions and AAO pore geometries. Substrate speeds and reactant pulse durations were defined by rotating cylinder rates of 10, 100, and 200 revolutions per minute (RPM). AAO pore diameters of 10, 25, 50, and 100 nm were utilized with a pore length of 25 μm. Uniform Zn coverage profiles were obtained at 10 RPM and pore diameters of 100 nm. The Zn coverage was less uniform at higher RPM values and smaller pore diameters. These results indicate that S-ALD into porous substrates is feasible under certain reaction conditions. S-ALD was then performed on porous Li ion battery electrodes to test S-ALD on a technologically important porous substrate. Li0.20Mn0.54Ni0.13Co0.13O2 electrodes on flexible metal foil were coated with Al2O3 using 2–5 Al2O3 ALD cycles. The Al2O3 ALD was performed in the S-ALD reactor at a rotating cylinder rate of 10 RPM using trimethylaluminum and ozone as the reactants at 50 °C. The capacity of the electrodes was then tested versus number of charge–discharge cycles. These measurements revealed that the Al2O3 S-ALD coating on the electrodes enhanced the capacity stability. This S-ALD process could be extended to roll-to-roll operation for the commercialization of S-ALD for coating Li ion battery electrodes.
Inherent substrate-dependent growth initiation and selective-area atomic layer deposition of TiO2 using “water-free” metal-halide/metal alkoxide reactants
Authors: Atanasov Sarah E. , Kalanyan Berç , Parsons Gregory N.
Published: December 2015

Titanium dioxide atomic layer deposition (ALD) is shown to proceed selectively on oxidized surfaces with minimal deposition on hydrogen-terminated silicon using titanium tetrachloride (TiCl4) and titanium tetra-isopropoxide [Ti(OCH(CH3)2)4, TTIP] precursors. Ex situ x-ray photoelectron spectroscopy shows a more rapid ALD nucleation rate on both Si–OH and Si–H surfaces when water is the oxygen source. Eliminating water delays the oxidation of the hydrogen-terminated silicon, thereby impeding TiO2 film growth. For deposition at 170 °C, the authors achieve ∼2 nm of TiO2 on SiO2 before substantial growth takes place on Si–H. On both Si–H and Si–OH, the surface reactions proceed during the first few TiCl4/TTIP ALD exposure steps where the resulting products act to impede subsequent growth, especially on Si–H surfaces. Insight from this work helps expand understanding of “inherent” substrate selective ALD, where native differences in substrate surface reaction chemistry are used to promote desired selective-area growth.
Probabilistic distributions of pinhole defects in atomic layer deposited films on polymeric substrates
Authors: Yersak Alexander S. , Lee Yung-Cheng
Published: December 2015

Pinhole defects in atomic layer deposition (ALD) coatings were measured in an area of 30 cm2 in an ALD reactor, and these defects were represented by a probabilistic cluster model instead of a single defect density value with number of defects over area. With the probabilistic cluster model, the pinhole defects were simulated over a manufacturing scale surface area of ∼1 m2. Large-area pinhole defect simulations were used to develop an improved and enhanced design method for ALD-based devices. A flexible thermal ground plane (FTGP) device requiring ALD hermetic coatings was used as an example. Using a single defect density value, it was determined that for an application with operation temperatures higher than 60 °C, the FTGP device would not be possible. The new probabilistic cluster model shows that up to 40.3% of the FTGP would be acceptable. With this new approach the manufacturing yield of ALD-enabled or other thin film based devices with different design configurations can be determined. It is important to guide process optimization and control and design for manufacturability.
Measuring two-phase particle flux with a multi-frequency acoustic Doppler profiler
Authors: Wilson Gregory W. , Hay Alex E.
Published: December 2015

A methodology is developed and tested for simultaneously extracting time-resolved one-dimensional profiles of the mass-concentration and velocity of two different particle types in a mixed suspension, using a multi-frequency pulse-to-pulse coherent Doppler instrument. The technique involves inversion of a model for frequency-dependent acoustic backscatter amplitude and phase. Results are presented from a laboratory settling column experiment, measuring a mixture of polystyrene beads (slowly-settling, strongly-scattering) and glass beads (quickly-settling, weakly-scattering) in a vertical pipe section.
New Commercially Available IgG Kits and Time-Resolved Fluorometric IgE Assay for Diagnosis of Allergic Bronchopulmonary Aspergillosis in Patients with Cystic Fibrosis
Authors: Barrera Coralie , Richaud-Thiriez Bénédicte , Rocchi Steffi , Rognon Bénédicte , Roussel Sandrine , Grenouillet Frédéric , Laboissière Audrey , Dalphin Jean-Charles , Reboux Gabriel , Millon Laurence
Published: December 2015

Allergic bronchopulmonary aspergillosis (ABPA) is difficult to diagnose; diagnosis relies on clinical, radiological, pathological, and serological criteria. Our aim was to assess the performance of two new commercially available kits and a new in-house assay: an
Aspergillus fumigatus
enzyme-linked immunosorbent assay (ELISA) IgG kit (Bordier Affinity Products), an
Aspergillus
Western blotting IgG kit (LDBio Diagnostics), and a new in-house time-resolved fluorometric IgE assay (dissociation-enhanced lanthanide fluorescent immunoassay, or DELFIA) using recombinant proteins from an
Aspergillus
sp. recently developed by our laboratory for ABPA diagnosis in a retrospective study that included 26 cystic fibrosis patients.
Aspergillus fumigatus
-specific IgG levels measured by a commercial ELISA kit were in accordance with the level of precipitins currently used in our lab. The ELISA kit could accelerate and help standardize ABPA diagnosis.
Aspergillus fumigatus
-specific IgE levels measured by ImmunoCAP (Phadia) with
A. fumigatus
M3 antigen and by DELFIA with a purified protein extract of
A. fumigatus
were significantly correlated (
P
< 10
−6
). The results with recombinant antigens glucose-6-phosphate isomerase and mannitol-1-phosphate dehydrogenase were encouraging but must be confirmed with sera from more patients. The DELFIA is an effective tool that can detect specific IgE against more fungal allergens than can be detected with other commercially available tests.
Draft Genome Sequence of
Arthrobacter
sp. Strain SPG23, a Hydrocarbon-Degrading and Plant Growth-Promoting Soil Bacterium
Authors: Gkorezis Panagiotis , Bottos Eric M. , Van Hamme Jonathan D. , Thijs Sofie , Rineau Francois , Franzetti Andrea , Balseiro-Romero Maria , Weyens Nele , Vangronsveld Jaco
Published: December 2015

We report here the 4.7-Mb draft genome of
Arthrobacter
sp. SPG23, a hydrocarbonoclastic Gram-positive bacterium belonging to the
Actinobacteria
, isolated from diesel-contaminated soil at the Ford Motor Company site in Genk, Belgium. Strain SPG23 is a potent plant growth promoter useful for diesel fuel remediation applications based on plant-bacterium associations.
Complete Genome Sequence of a Tomato-Infecting Tomato Mottle Mosaic Virus in New York
Authors: Padmanabhan Chellappan , Zheng Yi , Li Rugang , Martin Gregory B. , Fei Zhangjun , Ling Kai-Shu
Published: December 2015

The complete genome sequence of an isolate of tomato mottle mosaic virus (ToMMV) infecting tomatoes in New York was obtained using small RNA (sRNA) deep sequencing. ToMMV_NY-13 shared 99% sequence identity with isolates from Mexico and Florida. Broader distribution of this emerging virus is a cause for concern to the tomato industry.
Utility of Whole-Genome Sequencing in Characterizing Acinetobacter Epidemiology and Analyzing Hospital Outbreaks
Authors: Fitzpatrick Margaret A. , Ozer Egon A. , Hauser Alan R.
Published: December 2015

Acinetobacter baumannii
frequently causes nosocomial infections and outbreaks. Whole-genome sequencing (WGS) is a promising technique for strain typing and outbreak investigations. We compared the performance of conventional methods with WGS for strain typing clinical
Acinetobacter
isolates and analyzing a carbapenem-resistant
A. baumannii
(CRAB) outbreak. We performed two band-based typing techniques (pulsed-field gel electrophoresis and repetitive extragenic palindromic-PCR), multilocus sequence type (MLST) analysis, and WGS on 148
Acinetobacter calcoaceticus-A. baumannii
complex bloodstream isolates collected from a single hospital from 2005 to 2012. Phylogenetic trees inferred from core-genome single nucleotide polymorphisms (SNPs) confirmed three
Acinetobacter
species within this collection. Four major
A. baumannii
clonal lineages (as defined by MLST) circulated during the study, three of which are globally distributed and one of which is novel. WGS indicated that a threshold of 2,500 core SNPs accurately distinguished
A. baumannii
isolates from different clonal lineages. The band-based techniques performed poorly in assigning isolates to clonal lineages and exhibited little agreement with sequence-based techniques. After applying WGS to a CRAB outbreak that occurred during the study, we identified a threshold of 2.5 core SNPs that distinguished nonoutbreak from outbreak strains. WGS was more discriminatory than the band-based techniques and was used to construct a more accurate transmission map that resolved many of the plausible transmission routes suggested by epidemiologic links. Our study demonstrates that WGS is superior to conventional techniques for
A. baumannii
strain typing and outbreak analysis. These findings support the incorporation of WGS into health care infection prevention efforts.
Two Listeria monocytogenes Pseudo-outbreaks Caused by Contaminated Laboratory Culture Media
Authors: Matanock Almea , Katz Lee S. , Jackson Kelly A. , Kucerova Zuzana , Conrad Amanda R. , Glover William A. , Nguyen Von , Mohr Marika C. , Marsden-Haug Nicola , Thompson Deborah , Dunn John R. , Stroika Steven , Melius Beth , Tarr Cheryl , Dietrich Stephen E. , Kao Annie S. , Kornstein Laura , Li Zhen , Maroufi Azarnoush , Marder Ellyn P. , Meyer Rebecca , Perez-Osorio Ailyn C. , Reddy Vasudha , Reporter Roshan , Carleton Heather , Tweeten Samantha , Waechter HaeNa , Yee Lisa M. , Wise Matthew E. , Davis Kim , Jackson Brendan R.
Published: December 2015

Listeriosis is a serious foodborne infection that disproportionately affects elderly adults, pregnant women, newborns, and immunocompromised individuals. Diagnosis is made by culturing
Listeria monocytogenes
from sterile body fluids or from products of conception. This report describes the investigations of two listeriosis pseudo-outbreaks caused by contaminated laboratory media made from sheep blood.
Culture-Based and Culture-Independent Bacteriologic Analysis of Cystic Fibrosis Respiratory Specimens
Authors: Mahboubi Mona A. , Carmody Lisa A. , Foster Bridget K. , Kalikin Linda M. , VanDevanter Donald R. , LiPuma John J.
Published: December 2015

Cystic fibrosis (CF) is characterized by chronic infection and inflammation of the airways.
In vitro
culture of select bacterial species from respiratory specimens has been used to guide antimicrobial therapy in CF for the past few decades. More recently, DNA sequence-based, culture-independent approaches have been used to assess CF airway microbiology, although the role that these methods will (or should) have in routine microbiologic analysis of CF respiratory specimens is unclear. We performed DNA sequence analyses to detect bacterial species in 945 CF sputum samples that had been previously analyzed by selective CF culture. We determined the concordance of results based on culture and sequence analysis, highlighting the comparison of the results for the most prevalent genera. Although overall prevalence rates were comparable between the two methods, results varied by genus. While sequence analysis was more likely to detect
Achromobacter
,
Stenotrophomonas
, and
Burkholderia
, it was less likely to detect
Staphylococcus
.
Streptococcus
spp. were rarely reported in culture results but were the most frequently detected species by sequence analysis. A variety of obligate and facultative anaerobic species, not reported by culture, was also detected with high prevalence by sequence analysis. Sequence analysis indicated that in a considerable proportion of samples, taxa not reported by selective culture constituted a relatively high proportion of the total bacterial load, suggesting that routine CF culture may underrepresent significant segments of the bacterial communities inhabiting CF airways.
Urine Galactomannan-to-Creatinine Ratio for Detection of Invasive Aspergillosis in Patients with Hematological Malignancies
Authors: Reischies Frederike M. J. , Raggam Reinhard B. , Prattes Juergen , Krause Robert , Eigl Susanne , List Agnes , Quehenberger Franz , Strenger Volker , Wölfler Albert , Hoenigl Martin
Published: December 2015

Galactomannan (GM) testing of urine specimens may provide important advantages, compared to serum testing, such as easy noninvasive sample collection. We evaluated a total of 632 serial urine samples from 71 patients with underlying hematological malignancies and found that the urine GM/creatinine ratio, i.e., (urine GM level × 100)/urine creatinine level, which takes urine dilution into account, reliably detected invasive aspergillosis and may be a promising diagnostic tool for patients with hematological malignancies. (This study has been registered at ClinicalTrials.gov under registration no. NCT01576653.)
The Interferon-Stimulated Gene
Ifi27l2a
Restricts West Nile Virus Infection and Pathogenesis in a Cell-Type- and Region-Specific Manner
Authors: Lucas Tiffany M. , Richner Justin M. , Diamond Michael S.
Published: December 2015

The mammalian host responds to viral infections by inducing expression of hundreds of interferon-stimulated genes (ISGs). While the functional significance of many ISGs has yet to be determined, their cell type and temporal nature of expression suggest unique activities against specific pathogens. Using a combination of ectopic expression and gene silencing approaches in cell culture, we previously identified
Ifi27l2a
as a candidate antiviral ISG within neuronal subsets of the central nervous system (CNS) that restricts infection by West Nile virus (WNV), an encephalitic flavivirus of global concern. To investigate the physiological relevance of Ifi27l2a in the context of viral infection, we generated
Ifi27l2a
−/−
mice. Although adult mice lacking
Ifi27l2a
were more vulnerable to lethal WNV infection, the viral burden was greater only within the CNS, particularly in the brain stem, cerebellum, and spinal cord. Within neurons of the cerebellum and brain stem, in the context of WNV infection, a deficiency of Ifi27l2a was associated with less cell death, which likely contributed to sustained viral replication and higher titers in these regions. Infection studies in a primary cell culture revealed that
Ifi27l2a
−/−
cerebellar granule cell neurons and macrophages but not cerebral cortical neurons, embryonic fibroblasts, or dendritic cells sustained higher levels of WNV infection than wild-type cells and that this difference was greater under conditions of beta interferon (IFN-β) pretreatment. Collectively, these findings suggest that Ifi27l2a has an antiviral phenotype in subsets of cells and that at least some ISGs have specific inhibitory functions in restricted tissues.

IMPORTANCE
The interferon-stimulated
Ifi27l2a
gene is expressed differentially within the central nervous system upon interferon stimulation or viral infection. Prior studies in cell culture suggested an antiviral role for Ifi27l2a during infection by West Nile virus (WNV). To characterize its antiviral activity
in vivo
, we generated mice with a targeted gene deletion of
Ifi27l2a
. Based on extensive virological analyses, we determined that Ifi27l2a protects mice from WNV-induced mortality by contributing to the control of infection of the hindbrain and spinal cord, possibly by regulating cell death of neurons. This antiviral activity was validated in granule cell neurons derived from the cerebellum and in macrophages but was not observed in other cell types. Collectively, these data suggest that Ifi27l2a contributes to innate immune restriction of WNV in a cell-type- and tissue-specific manner.
Generation of Recombinant Oropouche Viruses Lacking the Nonstructural Protein NSm or NSs
Authors: Tilston-Lunel Natasha L. , Acrani Gustavo Olszanski , Randall Richard E. , Elliott Richard M.
Published: December 2015

Oropouche virus (OROV) is a midge-borne human pathogen with a geographic distribution in South America. OROV was first isolated in 1955, and since then, it has been known to cause recurring outbreaks of a dengue-like illness in the Amazonian regions of Brazil. OROV, however, remains one of the most poorly understood emerging viral zoonoses. Here we describe the successful recovery of infectious OROV entirely from cDNA copies of its genome and generation of OROV mutant viruses lacking either the NSm or the NSs coding region. Characterization of the recombinant viruses carried out
in vitro
demonstrated that the NSs protein of OROV is an interferon (IFN) antagonist as in other NSs-encoding bunyaviruses. Additionally, we demonstrate the importance of the nine C-terminal amino acids of OROV NSs in IFN antagonistic activity. OROV was also found to be sensitive to IFN-α when cells were pretreated; however, the virus was still capable of replicating at doses as high as 10,000 U/ml of IFN-α, in contrast to the family prototype BUNV. We found that OROV lacking the NSm protein displayed characteristics similar to those of the wild-type virus, suggesting that the NSm protein is dispensable for virus replication in the mammalian and mosquito cell lines that were tested.

IMPORTANCE
Oropouche virus (OROV) is a public health threat in Central and South America, where it causes periodic outbreaks of dengue-like illness. In Brazil, OROV is the second most frequent cause of arboviral febrile illness after dengue virus, and with the current rates of urban expansion, more cases of this emerging viral zoonosis could occur. To better understand the molecular biology of OROV, we have successfully rescued the virus along with mutants. We have established that the C terminus of the NSs protein is important in interferon antagonism and that the NSm protein is dispensable for virus replication in cell culture. The tools described in this paper are important in terms of understanding this important yet neglected human pathogen.
Antigenic and Cryo-Electron Microscopy Structure Analysis of a Chimeric Sapovirus Capsid
Authors: Miyazaki Naoyuki , Taylor David W. , Hansman Grant S. , Murata Kazuyoshi
Published: December 2015

The capsid protein (VP1) of all caliciviruses forms an icosahedral particle with two principal domains, shell (S) and protruding (P) domains, which are connected via a flexible hinge region. The S domain forms a scaffold surrounding the nucleic acid, while the P domains form a homodimer that interacts with receptors. The P domain is further subdivided into two subdomains, termed P1 and P2. The P2 subdomain is likely an insertion in the P1 subdomain; consequently, the P domain is divided into the P1-1, P2, and P1-2 subdomains. In order to investigate capsid antigenicity, N-terminal (N-term)/S/P1-1 and P2/P1-2 were switched between two sapovirus genotypes GI.1 and GI.5. The chimeric VP1 constructs were expressed in insect cells and were shown to self-assemble into virus-like particles (VLPs) morphologically similar to the parental VLPs. Interestingly, the chimeric VLPs had higher levels of cross-reactivities to heterogeneous antisera than the parental VLPs. In order to better understand the antigenicity from a structural perspective, we determined an intermediate-resolution (8.5-Å) cryo-electron microscopy (cryo-EM) structure of a chimeric VLP and developed a VP1 homology model. The cryo-EM structure revealed that the P domain dimers were raised slightly (∼5 Å) above the S domain. The VP1 homology model allowed us predict the S domain (67–229) and P1-1 (229–280), P2 (281–447), and P1-2 (448–567) subdomains. Our results suggested that the raised P dimers might expose immunoreactive S/P1-1 subdomain epitopes. Consequently, the higher levels of cross-reactivities with the chimeric VLPs resulted from a combination of GI.1 and GI.5 epitopes.

IMPORTANCE
We developed sapovirus chimeric VP1 constructs and produced the chimeric VLPs in insect cells. We found that both chimeric VLPs had a higher level of cross-reactivity against heterogeneous VLP antisera than the parental VLPs. The cryo-EM structure of one chimeric VLP (Yokote/Mc114) was solved to 8.5-Å resolution. A homology model of the VP1 indicated for the first time the putative S and P (P1-1, P2, and P1-2) domains. The overall structure of Yokote/Mc114 contained features common among other caliciviruses. We showed that the P2 subdomain was mainly involved in the homodimeric interface, whereas a large gap between the P1 subdomains had fewer interactions.
Lipid Membranes Facilitate Conformational Changes Required for Reovirus Cell Entry
Authors: Snyder Anthony J. , Danthi Pranav
Published: December 2015

Cellular entry of nonenveloped and enveloped viruses is often accompanied by dramatic conformational changes within viral structural proteins. These rearrangements are triggered by a variety of mechanisms, such as low pH, virus-receptor interactions, and virus-host chaperone interactions. Reoviruses, a model system for entry of nonenveloped viruses, undergo a series of disassembly steps within the host endosome. One of these steps, infectious subviral particle (ISVP)-to-ISVP* conversion, is necessary for delivering the genome-containing viral core into host cells, but the physiological trigger that mediates ISVP-to-ISVP* conversion during cell entry is unknown. Structural studies of the reovirus membrane penetration protein, μ1, predict that interactions between μ1 and negatively charged lipid head groups may promote ISVP* formation; however, experimental evidence for this idea is lacking. Here, we show that the presence of polyanions (SO
4
2−
and HPO
4
2−
) or lipids in the form of liposomes facilitates ISVP-to-ISVP* conversion. The requirement for charged lipids appears to be selective, since phosphatidylcholine and phosphatidylethanolamine promoted ISVP* formation, whereas other lipids, such as sphingomyelin and sulfatide, either did not affect ISVP* formation or prevented ISVP* formation. Thus, our work provides evidence that interactions with membranes can function as a trigger for a nonenveloped virus to gain entry into host cells.

IMPORTANCE
Cell entry, a critical stage in the virus life cycle, concludes with the delivery of the viral genetic material across host membranes. Regulated structural transitions within nonenveloped and enveloped viruses are necessary for accomplishing this step; these conformational changes are predominantly triggered by low pH and/or interactions with host proteins. In this work, we describe a previously unknown trigger, interactions with lipid membranes, which can induce the structural rearrangements required for cell entry. This mechanism operates during entry of mammalian orthoreoviruses. We show that interactions between reovirus entry intermediates and lipid membranes devoid of host proteins promote conformational changes within the viral outer capsid that lead to membrane penetration. Thus, this work illustrates a novel strategy that nonenveloped viruses can use to gain access into cells and how viruses usurp disparate host factors to initiate infection.
European Academies Advise on Gain-of-Function Studies in Influenza Virus Research
Authors: Fears Robin , ter Meulen Volker
Published: December 2015

Gain-of-function (GoF) studies to understand factors affecting transmissibility of potentially pandemic pathogens are controversial. The European Academies Science Advisory Council (EASAC) recently published consensus recommendations relating to GoF research review and management on self-regulation and harmonization; bioethical considerations; benefit-risk assessment; biosafety, and biosecurity advisory options; and publication of sensitive information. A layered approach to integration of responsibilities must include conforming to the stringent rules and guidance already existing. Further commitment is essential to extend the debate on issues worldwide.
Nocturnal haemoglobin oxygen desaturation in urban and rural East African paediatric cohorts with and without sickle cell anaemia: a cross-sectional study
Authors: L'Esperance VS , Ekong T , Cox SE , Makani J , Newton CR , Soka D , Komba A , Kirkham FJ , Hill CM
Published: December 2015

Low haemoglobin oxygen saturation (SpO2) predicts complications in children with sickle cell anaemia (SCA) in the North but there are few data from Africa, where the majority of the patients reside. We measured daytime and overnight SpO2 in children with SCA in routine follow-up clinic, and controls without symptoms of SCA, comparing rural (Kilifi, Kenya) and urban (Dar-es-Salaam, Tanzania) cohorts. Daytime SpO2 was lower in 65 Tanzanian children with SCA (TS; median 97 (IQR 94–100)%); p<0.0001) than in 113 Kenyan children with SCA (KS; 99 (98–100)%) and 20 Tanzanian controls (TC; 100 (98–100)%). Compared with 95 Kenyan children with SCA, in 54 Tanzanian children with SCA and 19 TC who returned for overnight oximetry, mean (KS 99.0 (96.7–99.8)%; TS 97.9 (95.4–99.3)%; TC 98.4 (97.5–99.1)%; p=0.01) and minimum nocturnal SpO2 (92 (86–95)%; 87 (78.5–91)%; 90 (83.5–93)% p=0.0001) were lower. The difference between children with SCA persisted after adjustment for haemoglobin (p=0.004). Urban Tanzanian children, with and without SCA, experience greater exposure to low daytime and night-time SpO2 compared with rural Kenyan children with SCA. Possible explanations include differences in the prevalence of obstructive sleep apnoea or asthma, alterations in the oxyhaemoglobin desaturation curve or cardiovascular compromise, for example, to shunting at atrial or pulmonary level secondary to increased pulmonary artery pressure. The fact that non-SCA siblings in the urban area are also affected suggests that environmental exposures, for example, air pollution, nutrition or physical exercise, may play a role. Further studies should determine aetiology and clinical relevance for the SCA phenotype in children resident in Africa.
Child Health General Practice Hubs: a service evaluation
Authors: Montgomery-Taylor Sarah , Watson Mando , Klaber Robert
Published: December 2015

Objective

To evaluate the impact of an integrated child health system.

Design

Mixed methods service evaluation.

Setting and patients

Children, young people and their families registered in Child Health General Practitioner (GP) Hubs where groups of GP practices come together to form ‘hubs’.

Interventions

Hospital paediatricians and GPs participating in joint clinics and multidisciplinary team (MDT) meetings in GP practices, a component of an ‘Inside-Out’ change known as ‘Connecting Care For Children (CC4C)’.

Main outcome measures

Cases seen in clinic or discussed at MDT meetings and their follow-up needs. Hospital Episode data: outpatient and inpatient activity and A&E attendance. Patient-reported experience measures and professionals’ feedback.

Results

In one hub, 39% of new patient hospital appointments were avoided altogether and a further 42% of appointments were shifted from hospital to GP practice. In addition, there was a 19% decrease in sub-specialty referrals, a 17% reduction in admissions and a 22% decrease in A&E attenders. Smaller hubs running at lower capacity in early stages of implementation had less impact on hospital activity. Patients preferred appointments at the GP practice, gained increased confidence in taking their child to the GP and all respondents said they would recommend the service to family and friends. Professionals valued the improvement in knowledge and learning and, most significantly, the development of trust and collaboration.

Conclusions

Child Health GP Hubs increase the connections between secondary and primary care, reduce secondary care usage and receive high patient satisfaction ratings while providing learning for professionals.
Exercise-based management versus surgery for multidirectional instability of the glenohumeral joint: a systematic review
Authors: Warby Sarah A , Pizzari Tania , Ford Jon J , Hahne Andrew J , Watson Lyn
Published: December 2015

Background

The most commonly recommended treatment for multidirectional instability (MDI) is exercise-based management, followed by surgery if exercise management fails. We aimed to evaluate the effectiveness of exercise-based management compared with surgery in patients with MDI. Secondary aims were to identify effective protocols and any adverse effects associated with exercise or surgery.

Method

The Cochrane database of systematic reviews, MEDLINE, EMBASE, CINAHL, PEDro, Current Contents, AMED, AMI, Ausport and Clinical Trials Registers were searched for published and unpublished studies from inception date to August 2014, using the key words multidirectional instability, and glenohumeral and exercise and surgery. Selection criteria included all study designs (except case reports and case series) and participants with MDI where both exercise-based management and surgery were compared in the same study. Inclusion criteria were not limited by outcomes. Risk of bias was assessed using Cochrane criteria. The GRADE approach was used to synthesise the evidence.

Results

4 non-randomised studies met the inclusion criteria. Risk of bias was high in all 4 studies. GRADE assessment revealed very low-quality evidence that surgery was superior to exercise therapy for impairment outcome measures, and exercise was favoured over surgery for patient-reported outcome measures.

The effect of exercise-based management compared with surgery for MDI is difficult to determine due to participant heterogeneity and a high level of bias across included studies.
Central pain processing is altered in people with Achilles tendinopathy
Authors: Tompra Nefeli , van Dieën Jaap H , Coppieters Michel W
Published: December 2015

Background

Tendinopathy is often a chronic condition. The mechanisms behind persistent tendon pain are not yet fully understood. It is unknown whether, similar to other persistent pain states, central pain mechanisms contribute to ongoing tendon pain.

Aim

We investigated the presence of altered central pain processing in Achilles tendinopathy by assessing the conditioned pain modulation (CPM) effect in people with and without Achilles tendinopathy.

Methods

20 people with Achilles tendinopathy and 23 healthy volunteers participated in this cross-sectional study. CPM was assessed by the cold pressor test. The pressure pain threshold (PPT) was recorded over the Achilles tendon before and during immersion of the participant's hand into cold water. The CPM effect was quantified as the absolute difference in PPT before and during the cold pressor test.

Results

An increase in PPT was observed in the Achilles tendinopathy and control group during the cold pressor test (p<0.001). However, the CPM effect was stronger in the control group (mean difference=160.5 kPa, SD=84.9 kPa) compared to the Achilles tendinopathy group (mean difference=36.4 kPa, SD=68.1 kPa; p<0.001).

We report a reduced conditioned pain modulation effect in people with Achilles tendinopathy compared to people without Achilles tendinopathy. A reduced conditioned pain modulation effect reflects altered central pain processing which is believed to contribute to the persistence of pain in other conditions. Altered central pain processing may also be an important factor in persistent tendon pain that has traditionally been regarded to be dominated by peripheral mechanisms.
RISUS study: Rugby Injury Surveillance in Ulster Schools
Authors: Archbold H A P , Rankin A T , Webb M , Nicholas R , Eames N W A , Wilson R K , Henderson L A , Heyes G J , Bleakley C M
Published: December 2015

Objective

To examine injury patterns in adolescent rugby players and determine factors associated with injury risk.

Design

Prospective injury surveillance study.

Setting

N=28 Grammar Schools in Ulster, Ireland (2014–2015 playing season).

Participants

825 adolescent rugby players, across in 28 school first XV rugby squads; mean age 16.9 years.

Main outcome measures

Injuries were classified by body part and diagnosis, and injury incidence using injuries per 1000 match hours of exposure. HRs for injury were calculated through Cox proportional hazard regression after correction for influential covariates.

Results

A total of n=426 injuries were reported across the playing season. Over 50% of injuries occurred in the tackle situation or during collisions (270/426), with few reported during set plays. The 3 most common injury sites were head/face (n=102, 23.9%), clavicle/shoulder (n=65, 15.3%) and the knee (n=56, 13.1%). Sprain (n=133, 31.2%), concussion (n=81, 19%) and muscle injury (n=65, 15.3%) were the most common diagnoses. Injury incidence is calculated at 29.06 injuries per 1000 match hours. There were no catastrophic injuries. A large percentage of injuries (208/424) resulted in absence from play for more than 28 days. Concussion carried the most significant time out from play (n=33; 15.9%), followed by dislocations of the shoulder (n=22; 10.6%), knee sprains (n=19, 9.1%), ankle sprains (n=14, 6.7%), hand/finger/thumb (n=11; 5.3%). 36.8% of participants in the study (304/825) suffered at least one injury during the playing season. Multivariate models found higher risk of injury (adjusted HR (AHR); 95% CI) with: higher age (AHR 1.45; 1.14 to 1.83), heavier weight (AHR 1.32; 1.04 to 1.69), playing representative rugby (AHR 1.42; 1.06 to 1.90) and undertaking regular strength training (AHR 1.65; 1.11 to 2.46). Playing for a lower ranked team (AHR 0.67; 0.49 to 0.90) and wearing a mouthguard (AHR 0.70; 0.54 to 0.92) were associated with lower risk of injury.

Conclusions

There was a high incidence of severe injuries, with concussion, ankle and knee ligament injuries and upper limb fractures/dislocations causing greatest time loss. Players were compliant with current graduated return-to-play regulations following concussion. Physical stature and levels of competition were important risk factors and there was limited evidence for protective equipment.
Death by homicide in National Collegiate Athletic Association athletes between 2003 and 2013
Authors: Rao Ashwin L , Poon Steven , Drezner Jonathan A , Zigman Monica , Asif Irfan M , Harmon Kimberly G
Published: December 2015

Background

The incidence of homicide-related death among individuals of college age in the United States population is estimated at 15.5/100 000. The incidence of homicide among National Collegiate Athletic Association (NCAA) athletes is unknown.

Aim

To investigate the rate of homicide-related death in NCAA athletes and to identify associated risk factors.

Methods

The NCAA Resolutions list, NCAA catastrophic insurance claims, media reports, and published NCAA demographic data were used to identify student athlete deaths and total participant seasons from 2003–04 through 2012–13. Homicide-related deaths were analysed by sex, race, division, sport, method, location, and circumstance. Internet searches were used to gather case details.

Results

Forty-two cases of homicide-related death were identified from 4 242 519 individual participant seasons during the ten-year study period. The incidence of homicide-related death in NCAA athletes was 1.0/100 000. The incidence in males was 1.45/100 000 and in females was 0.4/100 000 (relative risk (RR) 2.9, p=0.01). The incidence in black athletes was 4.2/100 000 and in white athletes was 0.4/100 000 (RR 7.0, p<0.001). The highest sport-specific homicide-related death rate was in American football (3.7/100 000), with a RR of 4.4 (p=0.002) compared to all other sports. 88% of cases occurred off-campus. 38% of cases occurred at a social gathering, and 38% of cases occurred in a place of residence. 74% involved a fatal shooting.

Conclusions

Homicide-related deaths in NCAA athletes occur most commonly in males, black athletes, and American football players. Understanding the incidence, risk factors, and circumstances of homicide-related deaths in college athletes may assist NCAA institutions in developing preventative measures.

Trial registration number

University of Washington Human Subjects Application, HSD No. 42077.
Qualitative study of telemonitoring of blood glucose and blood pressure in type 2 diabetes
Authors: Hanley Janet , Fairbrother Peter , McCloughan Lucy , Pagliari Claudia , Paterson Mary , Pinnock Hilary , Sheikh Aziz , Wild Sarah , McKinstry Brian
Published: December 2015

Objectives

To explore the experiences of patients and professionals taking part in a randomised controlled trial (RCT) of blood glucose, blood pressure (BP) and weight telemonitoring in type 2 diabetes supported by primary care, and identify factors facilitating or hindering the effectiveness of the intervention and those likely to influence its potential translation to routine practice.

Design

Qualitative study adopting an interpretive descriptive approach.

Participants

23 patients, 6 nurses and 4 doctors who were participating in a RCT of blood glucose and BP telemonitoring. A maximum variation sample of patients from within the trial based on age, sex and deprivation status of the practice was sought.

Setting

12 primary care practices in Scotland and England.

Method

Data were collected via recorded semistructured interviews. Analysis was inductive with themes presented within an overarching thematic framework. Multiple strategies were employed to ensure that the analysis was credible and trustworthy.

Results

Telemonitoring of blood glucose, BP and weight by people with type 2 diabetes was feasible. The data generated by telemonitoring supported self-care decisions and medical treatment decisions. Motivation to self-manage diet was increased by telemonitoring of blood glucose, and the ‘benign policing’ aspect of telemonitoring was considered by patients to be important. The convenience of home monitoring was very acceptable to patients although professionals had some concerns about telemonitoring increasing workload and costs.

Conclusions

Telemonitoring of blood glucose, BP and weight in primary care is a promising way of improving diabetes management which would be highly acceptable to the type of patients who volunteered for this study.

Trial registration number

ISRCTN71674628; Pre-results.