bluuebunny/crossref_metadata_2025 huggingface dataset:
The Environmental Impact Assessment of a Company’s Manufacturing System
Authors: Anagnostakis Dimitrios , Ritchie James M. , Lim Theodore , Craig Conor , Speedie Jamie
Published: January 2016

The major objective of this study is the estimation of the environmental impacts of a company’s manufacturing system. For this purpose, environmental key performance indicators are selected related to the energy consumption and pollutant gas emissions. To obtain accurate results and achieve an evaluation of the production system, a discrete event simulation tool was equipped. In this way, the production processes of a company were modelled and simulated in order to be assessed with regards to their environmental performance and impacts. The assessment of the investigated manufacturing system was carried out, by comparing it against a hypothetical ideal system of 100% efficiency in order for potential reductions in energy consumption and carbon emissions to be identified. Additionally, a supplementary use of the proposed methodology is presented showing that modelling the production system in advance a company can save energy and associated costs as well as reduce carbon emissions.
Robotic Neuromuscular Facilitation for Regaining Neural Activation in Hemiparetic Limbs
Authors: Ueda Jun , Lacey Lauren , Turkseven Melih , Shinohara Minoru , Kovalenko Ilya , Kim Euisun , Sulejmani Fatiesa
Published: January 2016

This paper introduces an effective engineered rehabilitation system for understanding and inducing functional recovery of hemiparetic limbs based on the concept of timing-dependent induction of neural plasticity. Limb motor function is commonly impaired after neurologic injury such as stroke, with hemiparesis being one of the major impairments. In an emerging unique intervention for hemiparesis, named repetitive facilitation exercise, or RFE, a therapist manually applies brief mechanical stimuli to the peripheral target muscles (e.g., tapping, stretching of tendon/muscle) immediately before a patient intends to produce a movement with the muscle. The practice of this rehabilitation procedure by a skilled therapist often leads to dramatic rehabilitation outcomes. However, unskilled therapists, most likely due to the inaccuracy of the timing of peripheral stimulation in reference to the intention of movement (i.e. motor command), are unable to recreate the same rehabilitation results. Robotic rehabilitation, on the other hand, can improve the reliability and efficacy of the operation by satisfying the timing precision required by the therapy. This study demonstrates the use of a pneumatically-driven MRI-compatible robot for RFE assessment. The pressure dynamics of the system is studied for an accurate estimation on the time of response of the robot. The required temporal precision of the therapy is obtained and the use of the device is validated through experiments on a human subject.
Numerical Simulation of Heavy Fuel Oil Combustion Characteristics and NOx Emissions in Calciner in Cement Industry
Authors: Abou ElSoad Hisham , Khalil Essam E.
Published: January 2016

The aim of the present study is to numerically investigate the combustion characteristics of Heavy Fuel Oil (HFO) and NOx emissions inside a Calciner used in cement industry. The combustion model was based on the conserved scalar (mixture fraction) and prescribed Probability Density Function (PDF) approach. The (RNG) k-ε turbulence model has been used. The HFO droplet trajectories were predicted by solving the momentum equations for the droplets using Lagrangian treatment. The radiation heat transfer equation was solved using P1 method. A swirl number greater than 0.6 was found to be optimal for good combustion characteristics and NOx emissions concentration. Meanwhile, it was found that the HFO viscosity value assumption has a significant effect on the injection velocity and must be considered as a function of temperature during the analysis as this will significantly affect the combustion characteristics.
Reducing Stress Concentration in RF MEMS Switch by Optimizing Serpentine Spring Design
Authors: Byron Reid , Hassanpour Pezhman
Published: January 2016

In this paper we present a method of reducing stress concentrations in serpentine beam structures to improve the fatigue life of RF MEMS switches. It was found that slight design modifications to the turning corners of serpentine springs can greatly improve the uniformity of stresses through the structure with minimal negative impact on the original spring constant. Before RF MEMS switches are widely adopted, they must achieve a low actuating voltage and working lives of greater than 10 billion cycles. The sharp turning corners of serpentine springs results in stress concentration creating an area prone to fatigue failure. In this paper, fillets of increasing radius were added to the inner and outer edges of a serpentine spring and the structure analyzed to determine their effect on global maximum stress and effective spring constant of the structure. It was found that adding fillets reduces maximum stress in the structure while redistributing the stress throughout the length of the meander. This results in a more uniform structure without significantly increasing the overall spring constant of the structure.
CXCR1-mediated neutrophil degranulation and fungal killing promote
Candida
clearance and host survival
Authors: Swamydas Muthulekha , Gao Ji-Liang , Break Timothy J. , Johnson Melissa D. , Jaeger Martin , Rodriguez Carlos A. , Lim Jean K. , Green Nathaniel M. , Collar Amanda L. , Fischer Brett G. , Lee Chyi-Chia Richard , Perfect John R. , Alexander Barbara D. , Kullberg Bart-Jan , Netea Mihai G. , Murphy Philip M. , Lionakis Michail S.
Published: January 2016

CXCR1 as a mediator of neutrophil fungal killing and host defense against systemic fungal infection in mice and humans.
Quantifying prion disease penetrance using large population control cohorts
Authors: Minikel Eric Vallabh , Vallabh Sonia M. , Lek Monkol , Estrada Karol , Samocha Kaitlin E. , Sathirapongsasuti J. Fah , McLean Cory Y. , Tung Joyce Y. , Yu Linda P. C. , Gambetti Pierluigi , Blevins Janis , Zhang Shulin , Cohen Yvonne , Chen Wei , Yamada Masahito , Hamaguchi Tsuyoshi , Sanjo Nobuo , Mizusawa Hidehiro , Nakamura Yosikazu , Kitamoto Tetsuyuki , Collins Steven J. , Boyd Alison , Will Robert G. , Knight Richard , Ponto Claudia , Zerr Inga , Kraus Theo F. J. , Eigenbrod Sabina , Giese Armin , Calero Miguel , de Pedro-Cuesta Jesús , Haïk Stéphane , Laplanche Jean-Louis , Bouaziz-Amar Elodie , Brandel Jean-Philippe , Capellari Sabina , Parchi Piero , Poleggi Anna , Ladogana Anna , O’Donnell-Luria Anne H. , Karczewski Konrad J. , Marshall Jamie L. , Boehnke Michael , Laakso Markku , Mohlke Karen L. , Kähler Anna , Chambert Kimberly , McCarroll Steven , Sullivan Patrick F. , Hultman Christina M. , Purcell Shaun M. , Sklar Pamela , van der Lee Sven J. , Rozemuller Annemieke , Jansen Casper , Hofman Albert , Kraaij Robert , van Rooij Jeroen G. J. , Ikram M. Arfan , Uitterlinden André G. , van Duijn Cornelia M. ,   Exome Aggregation Consortium (ExAC), Daly Mark J. , MacArthur Daniel G.
Published: January 2016

Large genomic reference data sets reveal a spectrum of pathogenicity in the prion protein gene and provide genetic validation for a therapeutic strategy in prion disease.
Host gene expression classifiers diagnose acute respiratory illness etiology
Authors: Tsalik Ephraim L. , Henao Ricardo , Nichols Marshall , Burke Thomas , Ko Emily R. , McClain Micah T. , Hudson Lori L. , Mazur Anna , Freeman Debra H. , Veldman Tim , Langley Raymond J. , Quackenbush Eugenia B. , Glickman Seth W. , Cairns Charles B. , Jaehne Anja K. , Rivers Emanuel P. , Otero Ronny M. , Zaas Aimee K. , Kingsmore Stephen F. , Lucas Joseph , Fowler Vance G. , Carin Lawrence , Ginsburg Geoffrey S. , Woods Christopher W.
Published: January 2016

Pathogen-specific host gene expression changes may combat inappropriate antibiotic use and emerging antibiotic resistance.
“Big data” gets personal
Authors: Lebo Matthew S. , Sutti Sheila , Green Robert C.
Published: January 2016

A woman at-risk for a rare fatal prion disease joined with a bioinformatics team to analyze shared data that provided insights about penetrance and treatment.
A T cell memory problem?
Authors: Dronca Roxana
Published: January 2016

Memory T cells promote functional differentiation of naïve T cells and impair the antitumor efficacy of adoptive immunotherapy.
Mutations in
gyrA
and
gyrB
among Fluoroquinolone- and Multidrug-Resistant Mycobacterium tuberculosis Isolates
Authors: Chien Jung-Yien , Chiu Wei-Yih , Chien Shun-Tien , Chiang Chia-Jung , Yu Chong-Jen , Hsueh Po-Ren
Published: January 2016

In order to correlate the mutations inside the entire
gyrA
and
gyrB
genes with the level of resistance to ofloxacin (OFX) and moxifloxacin (MFX) in isolates of multidrug-resistant
Mycobacterium tuberculosis
(MDR-TB), a total of 111 isolates were categorized into OFX-susceptible (MIC, ≤2 μg/ml) and low-level (MIC, 4 to 8 μg/ml) and high-level (MIC, ≥16 μg/ml) OFX-resistant isolates and MFX-susceptible (MIC, ≤0.5 μg/ml) and low-level (MIC, 1 to 2 μg/ml) and high-level (MIC, ≥4 μg/ml) MFX-resistant isolates. Resistance-associated mutations inside the
gyrA
gene were found in 30.2% of OFX-susceptible and 72.5% and 72.2% of low-level and high-level OFX-resistant isolates and in 28.6% of MFX-susceptible and 58.1% and 83.9% of low-level and high-level MFX-resistant isolates. Compared with OFX-susceptible isolates, low-level and high-level OFX-resistant isolates had a significantly higher prevalence of mutations at
gyrA
codons 88 to 94 (17.0%, 65.0%, and 72.2%, respectively;
P
< 0.001) and a higher prevalence of the
gyrB
G512R mutation (0.0%, 2.5%, and 16.7%, respectively;
P
= 0.006). Similarly, compared with MFX-susceptible isolates, low-level and high-level MFX-resistant isolates had a significantly higher prevalence of mutations at
gyrA
codons 88 to 94 (14.3%, 51.6%, and 80.6%, respectively;
P
< 0.001) as well as a higher prevalence of the
gyrB
G512R mutation (0.0%, 0.0%, and 12.9%, respectively;
P
= 0.011). D94G and D94N mutations in
gyrA
and the G512R mutation in
gyrB
were correlated with high-level MFX resistance, while the D94A mutation was associated with low-level MFX resistance. The prevalence of mutations at
gyrA
codons 88 to 94 and the
gyrB
G512R mutation were higher among fluoroquinolone (FQ)-susceptible East Asian (Beijing) and Indo-Oceanic strains than they were among Euro-American strains, implying that molecular techniques to detect FQ resistance may be less specific in areas with a high prevalence of East Asian (Beijing) and Indo-Oceanic strains.
Intra- and Interspecies Effects of Outer Membrane Vesicles from Stenotrophomonas maltophilia on β-Lactam Resistance
Authors: Devos Simon , Stremersch Stephan , Raemdonck Koen , Braeckmans Kevin , Devreese Bart
Published: January 2016

The treatment of
Stenotrophomonas maltophilia
infection with β-lactam antibiotics leads to increased release of outer membrane vesicles (OMVs), which are packed with two chromosomally encoded β-lactamases. Here, we show that these β-lactamase–packed OMVs are capable of establishing extracellular β-lactam degradation. We also show that they dramatically increase the apparent MICs of imipenem and ticarcillin for the cohabituating species
Pseudomonas aeruginosa
and
Burkholderia cenocepacia
.
Needle-Free Delivery of Acetalated Dextran-Encapsulated AR-12 Protects Mice from Francisella tularensis Lethal Challenge
Authors: Hoang Ky V. , Curry Heather , Collier Michael A. , Borteh Hassan , Bachelder Eric M. , Schlesinger Larry S. , Gunn John S. , Ainslie Kristy M.
Published: January 2016

Francisella tularensiscauses tularemia and is a potential biothreat. Given the limited antibiotics for treating tularemia and the possible use of antibiotic-resistant strains as a biowarfare agent, new antibacterial agents are needed. AR-12 is an FDA-approved investigational new drug (IND) compound that induces autophagy and has shown host-directed, broad-spectrum activityin vitroagainstSalmonella entericaserovar Typhimurium andF. tularensis. We have shown that AR-12 encapsulated within acetalated dextran (Ace-DEX) microparticles (AR-12/MPs) significantly reduces host cell cytotoxicity compared to that with free AR-12, while retaining the ability to controlS.Typhimurium within infected human macrophages. In the present study, the toxicity and efficacy of AR-12/MPs in controlling virulent type AF. tularensisSchuS4 infection were examinedin vitroandin vivo. No significant toxicity of blank MPs or AR-12/MPs was observed in lung histology sections when the formulations were given intranasally to uninfected mice. In histology sections from the lungs of intranasally infected mice treated with the formulations, increased macrophage infiltration was observed for AR-12/MPs, with or without suboptimal gentamicin treatment, but not for blank MPs, soluble AR-12, or suboptimal gentamicin alone. AR-12/MPs dramatically reduced the burden ofF. tularensisin infected human macrophages, in a manner similar to that of free AR-12. However,in vivo, AR-12/MPs significantly enhanced the survival ofF. tularensisSchuS4-infected mice compared to that seen with free AR-12. In combination with suboptimal gentamicin treatment, AR-12/MPs further improved the survival ofF. tularensisSchuS4-infected mice. These studies provide support for Ace-DEX-encapsulated AR-12 as a promising new therapeutic agent for tularemia.
Roles of Toll-Like Receptor 2 (TLR2), TLR4, and MyD88 during Pulmonary Coxiella burnetii Infection
Authors: Ramstead Andrew G. , Robison Amanda , Blackwell Anne , Jerome Maria , Freedman Brett , Lubick Kirk J. , Hedges Jodi F. , Jutila Mark A.
Published: January 2016

Coxiella burnetii, the causative agent of Q fever, is an obligate intracellular, primarily pulmonary, bacterial pathogen. Although much is known about adaptive immune responses against this bacterium, our understanding of innate immune responses againstC. burnetiiis not well defined, particularly within the target tissue for infection, the lung. Previous studies examined the roles of the innate immune system receptors Toll-like receptor 2 (TLR2) and TLR4 in peripheral infection models and described minimal phenotypes in specific gene deletion animals compared to those of their wild-type controls (S. Meghari et al., Ann N Y Acad Sci 1063:161–166, 2005,http://dx.doi.org/10.1196/annals.1355.025; A. Honstettre et al., J Immunol 172:3695–3703, 2004,http://dx.doi.org/10.4049/jimmunol.172.6.3695) . Here, we assessed the roles for TLR2, TLR4, and MyD88 in pulmonaryC. burnetiiinfection and compared responses to those that occurred in TLR2- and TLR4-deficient animals following peripheral infection. As observed previously, neither TLR2 nor TLR4 was needed for limiting bacterial growth after peripheral infection. In contrast, TLR2 and, to a lesser extent, TLR4 limited growth (or dissemination) of the bacterium in the lung and spleen after pulmonary infection. TLR2, TLR4, and MyD88 were not required for the general inflammatory response in the lungs after pulmonary infection. However, MyD88 signaling was important for infection-induced morbidity. Finally, TLR2 expression on hematopoietic cells was most important for limiting bacterial growth in the lung. These results expand on our knowledge of the roles for TLR2 and TLR4 inC. burnetiiinfection and suggest various roles for these receptors that are dictated by the site of infection.
Exploration of Sulfur Assimilation of Aspergillus fumigatus Reveals Biosynthesis of Sulfur-Containing Amino Acids as a Virulence Determinant
Authors: Amich Jorge , Dümig Michaela , O'Keeffe Gráinne , Binder Jasmin , Doyle Sean , Beilhack Andreas , Krappmann Sven
Published: January 2016

Fungal infections are of major relevance due to the increased numbers of immunocompromised patients, frequently delayed diagnosis, and limited therapeutics. To date, the growth and nutritional requirements of fungi during infection, which are relevant for invasion of the host, are poorly understood. This is particularly true for invasive pulmonary aspergillosis, as so far, sources of (macro)elements that are exploited during infection have been identified to only a limited extent. Here, we have investigated sulfur (S) utilization by the human-pathogenic mold
Aspergillus fumigatus
during invasive growth. Our data reveal that inorganic S compounds or taurine is unlikely to serve as an S source during invasive pulmonary aspergillosis since a sulfate transporter mutant strain and a sulfite reductase mutant strain are fully virulent. In contrast, the S-containing amino acid cysteine is limiting for fungal growth, as proven by the reduced virulence of a cysteine auxotroph. Moreover, phenotypic characterization of this strain further revealed the robustness of the subordinate glutathione redox system. Interestingly, we demonstrate that methionine synthase is essential for
A. fumigatus
virulence, defining the biosynthetic route of this proteinogenic amino acid as a potential antifungal target. In conclusion, we provide novel insights into the nutritional requirements of
A. fumigatus
during pathogenesis, a prerequisite to understanding and fighting infection.
Development of RNA Interference Trigger-Mediated Gene Silencing in Entamoeba invadens
Authors: Suresh Susmitha , Ehrenkaufer Gretchen , Zhang Hanbang , Singh Upinder
Published: January 2016

Entamoeba histolytica
, a protozoan parasite, is an important human pathogen and a leading parasitic cause of death. The organism has two life cycle stages, trophozoites, which are responsible for tissue invasion, and cysts, which are involved in pathogen transmission.
Entamoeba invadens
is the model system to study
Entamoeba
developmental biology, as high-grade regulated encystation and excystation are readily achievable. However, the lack of gene-silencing tools in
E. invadens
has limited the molecular studies that can be performed. Using the endogenous RNA interference (RNAi) pathway in
Entamoeba
, we developed an RNAi-based trigger gene-silencing approach in
E. invadens
. We demonstrate that a gene's coding region that has abundant antisense small RNAs (sRNAs) can trigger silencing of a gene that is fused to it. The trigger fusion leads to the generation of abundant antisense sRNAs that map to the target gene, with silencing occurring independently of trigger location at the 5′ or 3′ end of a gene. Gene silencing is stably maintained during development, including encystation and excystation. We have used this approach to successfully silence two
E. invadens
genes: a putative rhomboid protease gene and a SHAQKY family Myb gene. The Myb gene is upregulated during oxidative stress and development, and its downregulation led, as predicted, to decreased viability under oxidative stress and decreased cyst formation. Thus, the RNAi trigger silencing method can be used to successfully investigate the molecular functions of genes in
E. invadens
. Dissection of the molecular basis of
Entamoeba
stage conversion is now possible, representing an important technical advance for the system.
Streptococcus pneumoniae Colonization Disrupts the Microbial Community within the Upper Respiratory Tract of Aging Mice
Authors: Thevaranjan Netusha , Whelan Fiona J. , Puchta Alicja , Ashu Eta , Rossi Laura , Surette Michael G. , Bowdish Dawn M. E.
Published: January 2016

Nasopharyngeal colonization by the Gram-positive bacteriumStreptococcus pneumoniaeis a prerequisite for pneumonia and invasive pneumococcal diseases. Colonization is asymptomatic, involving dynamic and complex interplay between commensals, the host immune system, and environmental factors. The elderly are at an increased risk of developing pneumonia, which might be due to changes in the respiratory microbiota that would impact bacterial colonization and persistence within this niche. We hypothesized that the composition of the upper respiratory tract (URT) microbiota changes with age and subsequently can contribute to sustained colonization and inefficient clearance ofS. pneumoniae. To test this, we used a mouse model of pneumococcal colonization to compare the composition of the URT microbiota in young, middle-aged, and old mice in the naive state and during the course of colonization using nasal pharyngeal washes. Sequencing of variable region 3 (V3) of the 16S rRNA gene was used to identify changes occurring with age and throughout the course ofS. pneumoniaecolonization. We discovered that age affects the composition of the URT microbiota and that colonization withS. pneumoniaeis more disruptive of preexisting communities in older mice. We have further shown that host-pathogen interactions followingS. pneumoniaecolonization can impact the populations of resident microbes, includingStaphylococcusandHaemophilus. Together, our findings indicate alterations to the URT microbiota could be detrimental to the elderly, resulting in increased colonization ofS. pneumoniaeand decreased efficiency in its clearance.
ThefruRBAOperon Is Necessary for Group A Streptococcal Growth in Fructose and for Resistance to Neutrophil Killing during Growth in Whole Human Blood
Authors: Valdes Kayla M. , Sundar Ganesh S. , Vega Luis A. , Belew Ashton T. , Islam Emrul , Binet Rachel , El-Sayed Najib M. , Le Breton Yoann , McIver Kevin S.
Published: January 2016

Bacterial pathogens rely on the availability of nutrients for survival in the host environment. The phosphoenolpyruvate-phosphotransferase system (PTS) is a global regulatory network connecting sugar uptake with signal transduction. Since the fructose PTS has been shown to impact virulence in several streptococci, including the human pathogenStreptococcus pyogenes(the group AStreptococcus[GAS]), we characterized its role in carbon metabolism and pathogenesis in the M1T1 strain 5448. Growth in fructose as a sole carbon source resulted in 103 genes affected transcriptionally, where thefrulocus (fruRBA) was the most induced. Reverse transcriptase PCR showed thatfruRBAformed an operon which was repressed by FruR in the absence of fructose, in addition to being under carbon catabolic repression. Growth assays and carbon utilization profiles revealed that although the entirefruoperon was required for growth in fructose, FruA was the main transporter for fructose and also was involved in the utilization of three additional PTS sugars: cellobiose, mannitol, andN-acetyl-d-galactosamine. The inactivation ofsloR, afruAhomolog that also was upregulated in the presence of fructose, failed to reveal a role as a secondary fructose transporter. Whereas the ability of both ΔfruRand ΔfruBmutants to survive in the presence of whole human blood or neutrophils was impaired, the phenotype was not reproduced in murine whole blood, and those mutants were not attenuated in a mouse intraperitoneal infection. Since the ΔfruAmutant exhibited no phenotype in the human or mouse assays, we propose that FruR and FruB are important for GAS survival in a human-specific environment.
Tissue-Resident T Cells as the Central Paradigm of Chlamydia Immunity
Authors: Johnson Raymond M. , Brunham Robert C.
Published: January 2016

For almost 2 decades, results fromChlamydiapathogenesis investigations have been conceptualized using a cytokine polarization narrative. Recent viral immunity studies identifying protective tissue-resident memory T cells (Trm) suggest an alternative paradigm based on localized immune networks. AsChlamydiavaccines enter the preclinical pipeline and, in the case of an attenuated trachoma vaccine, are given to human subjects, it may be useful to ask whether cytokine polarization is the appropriate framework for understanding and evaluating vaccine efficacy. In this review, we revisitC. trachomatispathogenesis data from mice and humans using a Trm narrative and note a comfortable concordance with theChlamydiapathogenesis literature.
Genes Required for the Fitness of Salmonella enterica Serovar Typhimurium during Infection of Immunodeficientgp91−/−phoxMice
Authors: Grant Andrew J. , Oshota Olusegun , Chaudhuri Roy R. , Mayho Matthew , Peters Sarah E. , Clare Simon , Maskell Duncan J. , Mastroeni Pietro
Published: January 2016

Salmonella entericacauses systemic diseases (typhoid and paratyphoid fever), nontyphoidal septicemia (NTS), and gastroenteritis in humans and other animals worldwide. An important but underrecognized emerging infectious disease problem in sub-Saharan Africa is NTS in children and immunocompromised adults. A current goal is to identifySalmonellamutants that are not pathogenic in the absence of key components of the immune system such as might be found in immunocompromised hosts. Such attenuated strains have the potential to be used as live vaccines. We have used transposon-directed insertion site sequencing (TraDIS) to screen mutants ofSalmonella entericaserovar Typhimurium for their ability to infect and grow in the tissues of wild-type and immunodeficient mice. This was to identify bacterial genes that might be deleted for the development of live attenuated vaccines that would be safer to use in situations and/or geographical areas where immunodeficiencies are prevalent. The relative fitness of each of 9,356 transposon mutants, representing mutations in 3,139 different genes, was determined ingp91−/−phoxmice. Mutations in certain genes led to reduced fitness in both wild-type and mutant mice. To validate these results, these genes were mutated by allelic replacement, and resultant mutants were retested for fitness in the mice. A defined deletion mutant ofcysEwas attenuated in C57BL/6 wild-type mice and immunodeficientgp91−/−phoxmice and was effective as a live vaccine in wild-type mice.
Deficiency of the Complement Component 3 but Not Factor B Aggravates Staphylococcus aureus Septic Arthritis in Mice
Authors: Na Manli , Jarneborn Anders , Ali Abukar , Welin Amanda , Magnusson Malin , Stokowska Anna , Pekna Marcela , Jin Tao
Published: January 2016

The complement system plays an essential role in the innate immune response and protection against bacterial infections. However, detailed knowledge regarding the role of complement inStaphylococcus aureusseptic arthritis is still largely missing. In this study, we elucidated the roles of selected complement proteins inS. aureusseptic arthritis. Mice lacking the complement component 3 (C3−/−), complement factor B (fB−/−), and receptor for C3-derived anaphylatoxin C3a (C3aR−/−) and wild-type (WT) control mice were intravenously or intra-articularly inoculated withS. aureusstrain Newman. The clinical course of septic arthritis, as well as histopathological and radiological changes in joints, was assessed. After intravenous inoculation, arthritis severity and frequency were significantly higher inC3−/−mice than in WT controls, whereasfB−/−mice displayed intermediate arthritis severity and frequency. This was in accordance with both histopathological and radiological findings. C3, but not fB, deficiency was associated with greater weight loss, more frequent kidney abscesses, and higher bacterial burden in kidneys.S. aureusopsonized withC3−/−sera displayed decreased uptake by mouse peritoneal macrophages compared with bacteria opsonized with WT orfB−/−sera. C3aR deficiency had no effect on the course of hematogenousS. aureusseptic arthritis. We conclude that C3 deficiency increases susceptibility to hematogenousS. aureusseptic arthritis and impairs host bacterial clearance, conceivably due to diminished opsonization and phagocytosis ofS. aureus.
Sibling Rivalry in Myxococcus xanthus Is Mediated by Kin Recognition and a Polyploid Prophage
Authors: Dey Arup , Vassallo Christopher N. , Conklin Austin C. , Pathak Darshankumar T. , Troselj Vera , Wall Daniel
Published: January 2016

Myxobacteria form complex social communities that elicit multicellular behaviors. One such behavior is kin recognition, in which cells identify siblings via their polymorphic TraA cell surface receptor, to transiently fuse outer membranes and exchange their contents. In addition, outer membrane exchange (OME) regulates behaviors, such as inhibition of wild-type
Myxococcus xanthus
(DK1622) from swarming. Here we monitored the fate of motile cells and surprisingly found they were killed by nonmotile siblings. The kill phenotype required OME (i.e., was TraA dependent). The genetic basis of killing was traced to ancestral strains used to construct DK1622. Specifically, the kill phenotype mapped to a large “polyploid prophage,” Mx alpha. Sensitive strains contained a 200-kb deletion that removed two of three Mx alpha units. To explain these results, we suggest that Mx alpha expresses a toxin-antitoxin cassette that uses the OME machinery of
M. xanthus
to transfer a toxin that makes the population “addicted” to Mx alpha. Thus, siblings that lost Mx alpha units (no immunity) are killed by cells that harbor the element. To test this, an Mx alpha-harboring laboratory strain was engineered (by
traA
allele swap) to recognize a closely related species,
Myxococcus fulvus
. As a result,
M. fulvus
, which lacks Mx alpha, was killed. These TraA-mediated antagonisms provide an explanation for how kin recognition specificity might have evolved in myxobacteria. That is, recognition specificity is determined by polymorphisms in
traA
, which we hypothesize were selected for because OME with non-kin leads to lethal outcomes.

IMPORTANCE
The transition from single cell to multicellular life is considered a major evolutionary event. Myxobacteria have successfully made this transition. For example, in response to starvation, individual cells aggregate into multicellular fruiting bodies wherein cells differentiate into spores. To build fruits, cells need to recognize their siblings, and in part, this is mediated by the TraA cell surface receptor. Surprisingly, we report that TraA recognition can also involve sibling killing. We show that killing originates from a prophage-like element that has apparently hijacked the TraA system to deliver a toxin to kin. We hypothesize that this killing system has imposed selective pressures on kin recognition, which in turn has resulted in TraA polymorphisms and hence many different recognition groups.
Functional Determinants of Metal Ion Transport and Selectivity in Paralogous Cation Diffusion Facilitator Transporters CzcD and MntE in Streptococcus pneumoniae
Authors: Martin Julia E. , Giedroc David P.
Published: January 2016

Cation diffusion facilitators (CDFs) are a large family of divalent metal transporters that collectively possess broad metal specificity and contribute to intracellular metal homeostasis and virulence in bacterial pathogens.
Streptococcus pneumoniae
expresses two homologous CDF efflux transporters, MntE and CzcD. Cells lacking
mntE
or
czcD
are sensitive to manganese (Mn) or zinc (Zn) toxicity, respectively, and specifically accumulate Mn or Zn, respectively, thus suggesting that MntE selectively transports Mn, while CzcD transports Zn. Here, we probe the origin of this metal specificity using a phenotypic growth analysis of pneumococcal variants. Structural homology to
Escherichia coli
YiiP predicts that both MntE and CzcD are dimeric and each protomer harbors four pairs of conserved metal-binding sites, termed the A site, the B site, and the C1/C2 binuclear site. We find that single amino acid mutations within both the transmembrane domain A site and the B site in both CDFs result in a cellular metal sensitivity similar to that of the corresponding null mutants. However, multiple mutations in the predicted cytoplasmic C1/C2 cluster of MntE have no impact on cellular Mn resistance, in contrast to the analogous substitutions in CzcD, which do have on impact on cellular Zn resistance. Deletion of the MntE-specific C-terminal tail, present only in Mn-specific bacterial CDFs, resulted in only a modest growth phenotype. Further analysis of MntE-CzcD functional chimeric transporters showed that Asn and Asp in the
N
D-
D
D A-site motif of MntE and the most N-terminal His in the
H
D-HD site A of CzcD (the specified amino acids are underlined) play key roles in transporter metal selectivity.

IMPORTANCE
Cation diffusion facilitator (CDF) proteins are divalent metal ion transporters that are conserved in organisms ranging from bacteria to humans and that play important roles in cellular physiology, from metal homeostasis and resistance to type I diabetes in vertebrates. The respiratory pathogen
Streptococcus pneumoniae
expresses two metal CDF transporters, CzcD and MntE. How CDFs achieve metal selectivity is unclear. We show here that CzcD and MntE are true paralogs, as CzcD transports zinc, while MntE selectively transports manganese. Through the use of an extensive collection of pneumococcal variants, we show that a primary determinant for metal selectivity is the A site within the transmembrane domain. This extends our understanding of how CDFs discriminate among transition metals.
Identification of a Functional Plasmodesmal Localization Signal in a Plant Viral Cell-To-Cell-Movement Protein
Authors: Yuan Cheng , Lazarowitz Sondra G. , Citovsky Vitaly
Published: January 2016

Our fundamental knowledge of the protein-sorting pathways required for plant cell-to-cell trafficking and communication via the intercellular connections termed plasmodesmata has been severely limited by the paucity of plasmodesmal targeting sequences that have been identified to date. To address this limitation, we have identified the plasmodesmal localization signal (PLS) in the
Tobacco mosaic virus
(TMV) cell-to-cell-movement protein (MP), which has emerged as the paradigm for dissecting the molecular details of cell-to-cell transport through plasmodesmata. We report here the identification of a bona fide functional TMV MP PLS, which encompasses amino acid residues between positions 1 and 50, with residues Val-4 and Phe-14 potentially representing critical sites for PLS function that most likely affect protein conformation or protein interactions. We then demonstrated that this PLS is both necessary and sufficient for protein targeting to plasmodesmata. Importantly, as TMV MP traffics to plasmodesmata by a mechanism that is distinct from those of the three plant cell proteins in which PLSs have been reported, our findings provide important new insights to expand our understanding of protein-sorting pathways to plasmodesmata.

IMPORTANCE
The science of virology began with the discovery of
Tobacco mosaic virus
(TMV). Since then, TMV has served as an experimental and conceptual model for studies of viruses and dissection of virus-host interactions. Indeed, the TMV cell-to-cell-movement protein (MP) has emerged as the paradigm for dissecting the molecular details of cell-to-cell transport through the plant intercellular connections termed plasmodesmata. However, one of the most fundamental and key functional features of TMV MP, its putative plasmodesmal localization signal (PLS), has not been identified. Here, we fill this gap in our knowledge and identify the TMV MP PLS.
Sunshine and vitamin D
Authors: Saraff Vrinda , Shaw Nick
Published: January 2016

Vitamin D is vital for bone health and its deficiency deemed as a disease of the past has re-emerged as an important health concern. Exposure of the skin to solar ultraviolet B radiation is the major source of vitamin D and only a small proportion is derived from dietary intake. We review the various factors that influence the cutaneous synthesis of vitamin D and the recommendations regarding safe sun exposure and dietary supplementation to achieve adequate vitamin D levels proposed for optimal bone health.
Viral haemorrhagic fever in children
Authors: MacDermott Nathalie E , De Surjo , Herberg Jethro A
Published: January 2016

Viral haemorrhagic fevers (VHFs) are currently at the forefront of the world's attention due to the recentZaire ebolavirusepidemic in West Africa. This epidemic has highlighted the frailty of the world's public health response mechanisms and demonstrated the potential risks to nations around the world of imported cases of epidemic diseases. While imported cases in children are less likely, the potential for such a scenario remains. It is therefore essential that paediatricians are aware of and prepared for potential imported cases of tropical diseases, VHFs being of particular importance due to their propensity to cause nosocomial spread. Examining the four families of viruses—Filoviridae,Arenaviridae,BunyaviridaeandFlaviviridae, we describe the different types of VHFs, with emphasis on differentiation from other diseases through detailed history-taking, their presentation and management from a paediatric perspective.
Summative service and stakeholder evaluation of an NHS-funded community Pharmacy Emergency Repeat Medication Supply Service (PERMSS)
Authors: Nazar Hamde , Nazar Zachariah , Simpson Jill , Yeung Andre , Whittlesea Cate
Published: January 2016

Objectives

Service and stakeholder evaluation of an NHS-funded service providing out-ofhours (OOH) emergency repeat medications to patients self-presenting at community pharmacies.

Setting

Community pharmacies across the North East of England accredited to provide this service.

Participants

Patients self-presenting to community pharmacies during OOH periods with emergency repeat medication supply requests.

Intervention

Community pharmacists assessed each request for clinical appropriateness and when suitable provide an emergency repeat medication supply, with additional pharmaceutical advice and services if required.

Primary outcomes

Number of emergency repeat medication supplies, time of request, reason for access, medication(s), pharmaceutical advice and services provided. Secondary outcomes were community pharmacist and patient satisfaction.

Results

A total of 2485 patients were managed across 227 community pharmacies (15 December 2014 to 7 April 2015). Most patients presented on Saturdays, with increased activity over national holidays. Older age was associated with increased service use. Of the 3226 medications provided, 439 were classified as high risk. Patients found this service easy to access and were willing to access the community pharmacy in the future for medication-related issues. In the absence of this service, 50% of patients would have missed their medication(s) until they saw their doctor and a further 46% would have accessed an alternative service. The cost of National Health Service (NHS) service(s) for patients who would have accessed an alternative OOH service was estimated as 37 times that of the community pharmacy service provided. Community pharmacists were happy to provide this service despite increased consultation times and workload.

Conclusions

Community pharmacists were able to manage patients’ OOH requests for emergency repeat medication and patients were happy with the service provided. Since the service cost was favourable when compared with alternative OOH services, it would be a viable option to reduce the workload on the wider NHS.
Attitudes and beliefs about deceased organ donation in the Arabic-speaking community in Australia: a focus group study
Authors: Ralph Angelique F , Alyami Ali , Allen Richard D M , Howard Kirsten , Craig Jonathan C , Chadban Steve J , Irving Michelle , Tong Allison
Published: January 2016

Objectives

To describe the beliefs and attitudes to organ donation in the Arabic-speaking community.

Design

Arabic-speaking participants were purposively recruited to participate in 6 focus groups. Transcripts were analysed thematically.

Participants

53 participants, aged 19–77 years, and originating from 8 countries, participated in 1 of 6 focus groups. Participants identified as Christian (73%), Islam (26%), Buddhist (2%) or did not identify with any religion (2%).

Results

6 themes (with subthemes) were identified; religious conviction; invisibility of organ donation; medical suspicion; owning the decision; and reciprocal benefit.

Conclusions

Although organ donation is considered a generous life-saving ‘gift’, representative members of the Arabic-speaking community in Australia were unfamiliar with, unnerved by and sceptical about the donation process. Making positive decisions about organ donation would likely require resolving tensions between respecting family, community and religious values versus their individual autonomy. Providing targeted education about the process and benefits of organ donation within the Arabic community may clarify ambiguities surrounding cultural and religious-based views on organ donation, reduce taboos and suspicion towards donation, and in turn, lead to increased organ donation rates.
The sensitivity and specificity of clinical measures of sport concussion: three tests are better than one
Authors: Resch Jacob E , Brown Cathleen N , Schmidt Julianne , Macciocchi Stephen N , Blueitt Damond , Cullum C Munro , Ferrara Michael S
Published: January 2016

Context

A battery of clinical measures of neurocognition, balance and symptoms has been recommended for the management of sport concussion (SC) but is based on variable evidence.

Objective

To examine the sensitivity and specificity of a battery of tests to assess SC in college athletes.

Design

Cross-sectional.

Setting

Research laboratory.

Patients or other participants

Division 1 athletes diagnosed with a SC (n=40) who were 20.2±1.60 years of age and 180.5±11.12 cm tall and healthy athletes (n=40) who were 19.0±0.93 years of age and 179.1±11.39 cm tall were enrolled.

Intervention(s)

Participants were administered Immediate Postconcussion Assessment and Cognitive Test (ImPACT), the Sensory Organization Test (SOT) and the Revised Head Injury Scale (HIS-r) prior to and up to 24 h following injury between the 2004 and 2014 sport seasons. Sensitivity and specificity were calculated using predictive discriminant analyses (PDA) and clinical interpretation guidelines.

Main outcome measures

Outcome measures included baseline and postinjury ImPACT, SOT and HIS-r composite scores.

Results

Using PDA, each clinical measure's sensitivity ranged from 55.0% to 77.5% and specificity ranged from 52.5% to 100%. The test battery possessed a sensitivity and specificity of 80.0% and 100%, respectively. Using clinical interpretation guidelines, sensitivity ranged from 55% to 97.5% individually, and 100% when combined.

Conclusions

Our results support a multidimensional approach to assess SC in college athletes which correctly identified 80–100% of concussed participants as injured. When each test was evaluated separately, up to 47.5% of our sample was misclassified. Caution is warranted when using singular measures to manage SC.
We have the programme, what next? Planning the implementation of an injury prevention programme
Authors: Donaldson Alex , Lloyd David G , Gabbe Belinda J , Cook Jill , Finch Caroline F
Published: January 2016

Background and aim

The impact of any injury prevention programme is a function of the programme and its implementation. However, real world implementation of injury prevention programmes is challenging. Lower limb injuries (LLIs) are common in community Australian football (community-AF) and it is likely that many could be prevented by implementing exercise-based warm-up programmes for players. This paper describes a systematic, evidence-informed approach used to develop the implementation plan for a LLI prevention programme in community-AF in Victoria, Australia.

Methods

An ecological approach, using Step 5 of the Intervention Mapping health promotion programme planning protocol, was taken.

Results

An implementation advisory group was established to ensure the implementation plan and associated strategies were relevant to the local context. Coaches were identified as the primary programme adopters and implementers within an ecological system including players, other coaches, first-aid providers, and club and league administrators. Social Cognitive Theory was used to identify likely determinants of programme reach, adoption and implementation among coaches (eg, knowledge, beliefs, skills and environment). Diffusion of Innovations theory, the Implementation Drivers framework and available research evidence were used to identify potential implementation strategies including the use of multiple communication channels, programme resources, coach education and mentoring.

Conclusions

A strategic evidence-informed approach to implementing interventions will help maximise their population impact. The approach to implementation planning described in this study relied on an effective researcher-practitioner partnership and active engagement of stakeholders. The identified implementation strategies were informed by theory, evidence and an in-depth understanding of the implementation context.
Marked socioeconomic inequalities in hip fracture incidence rates during the Bone and Joint Decade (2000–2010) in Portugal: age and sex temporal trends in a population based study
Authors: Oliveira Carla Maria , Alves Sandra Maria , Pina Maria Fátima
Published: January 2016

Background

Socioeconomic factors may influence changes in hip fracture (HF) incidence over time. We analysed HF temporal trends during the Bone and Joint Decade in Portugal (BJD-Portugal), 2000–2010, by regional socioeconomic status (SES), sex and age.

Methods

We selected registers of patients aged 50+ years with HF (International Classification of Diseases, V.9—Clinical Modification, ICD9-CM) caused by traumas of low/moderate energy, from the National Hospital Discharge Database. Annual time series of age-specific incidence rates were calculated by sex and regional SES (deprived, medium, affluent). Generalised additive models were fitted to identify shape/turning points in temporal trends.

Results

We selected 96 905 HF (77.3% in women). Women were older than men at admission (81.2±8.5 vs 78.2±10.1 years-old, p<0.001). For women 65–79 years, a continuously decreasing trend (1.7%/year) only in affluent and increasing trends (3.3–3.4%/year) after 2006/2007 in medium and deprived was observed. For men, trends were stable or increased in almost all age/SES groups (only two decreasing periods). For the oldest women, all SES present similar trends: turning points around 2003 (initiating decreasing periods: 1.8–2.9%/year) and around 2007 (initiating increasing periods: 3.7–3.3%/year).

Conclusions

There were SES-sex-age inequalities in temporal trends during BJD-Portugal: marked SES inequalities among women aged 65–79 years (a persistent, decreasing trend only in the affluent) vanished among the oldest women; the same was not observed in men, for them, there were almost no declining periods; women aged ≥80 years, presented increasing trends around 2007, as in most deprived/age/sex groups. Despite some successful periods of decreasing trends, incidence rates did not improve overall in almost all age groups and both sexes.