bluuebunny/crossref_metadata_2025 huggingface dataset:
The Performance and Pedagogy of Migration in the Foreign Language Classroom: Staging Roland Schimmelpfennig's Der goldene Drache
Authors: Guenther Christina
Published: November 2017

This article explores how drama pedagogy in the foreign language classroom serves as a unique avenue for approaching the global issue of migration in the context of German‐speaking Europe. The article focuses on how staging a contemporary play about migration, such as Roland Schimmelpfennig's Der goldene Drache (2009), in German for an English‐speaking student and community audience can serve as a particularly rich example of cross‐cultural translation. The article outlines the various steps in experiential learning involved in a German‐language drama course and reflects on the potential for students and their instructor to be culturally transformed through the language of drama and the representation of migration. A brief discussion of Schimmelpfennig's play and its production in the context of a German drama course at a Midwestern university illustrates what the performance and pedagogy of migration might look like in the German foreign language classroom.
Cover Image, Volume 15, Issue 4
Authors: Lai S. R. , Castello S. A. , Robinson A. C. , Koehler J. W.
Published: November 2017

The cover image, by S. R. Lai et al., is based on the Original Article In vitro anti‐tubulin effects of mebendazole and fenbendazole on canine glioma cells, DOI: 10.1111/vco.12288

image
Standing thyroidectomy in 10 horses
Authors: Marcatili Marco , Voss Sarah J. , Pollock Patrick J.
Published: November 2017

Objective

To describe a surgical technique for thyroidectomy in horses with thyroid neoplasia under standing sedation and local anesthesia.

Study design

Retrospective study.

Animals

Client‐owned horses (n = 10).

Methods

Medical records of horses with a history of thyroid enlargement were included in the study if thyroid gland enlargement was treated surgically via hemi‐ or bilateral thyroidectomy, with the horse standing and sedated. Data derived from follow‐up clinical examination, performance level, recurrence, and cosmetic outcome were evaluated.

Results

Thyroid enlargement was unilateral in 8 and bilateral in 2 horses. Histopathological findings included adenomas (5/10), adenocarcinomas (2/10), cystic hyperplasia (2/10), and C‐cell adenoma (1/10). No major complications were encountered during or after surgery. All horses resumed their previous level of exercise within 6 weeks. Recurrence was diagnosed in 1 horse, 7 months after excision, and a second surgery was required. Recurrent laryngeal nerve neuropathy and seroma formation subsequent to surgery were not recorded in any of the cases.

Conclusion

Thyroidectomy can safely be performed with the horse standing and sedated with local anesthesia.

Clinical relevance

Performing standing thyroidectomy does not increase intraoperative or postoperative complications and could be considered for horses with thyroid enlargement amenable to surgery.
A Model-Free Continuous Velocity Observer Formulation With Self-Tuning for Mechatronic Systems
Authors: Deniz Meryem , Bayrak Alper , Tatlicioglu Enver , Zergeroglu Erkan
Published: November 2017

In this study, the design of a smooth robust velocity observer for a class of uncertain nonlinear mechatronic systems is presented. The proposed velocity observer does not require a priori knowledge of the upper bounds of the uncertain system dynamics and introduces time-varying observer gains for uncertainty compensation. Practical stability of the velocity observation error is ensured via Lyapunov-type stability analysis. Experimental results obtained from Phantom Omni haptic device are presented to illustrate the performance of the proposed velocity observer.
Consensus-Based Cooperative Formation Control for Multiquadcopter System With Unidirectional Network Connections
Authors: Namerikawa Toru , Kuriki Yasuhiro , Khalifa Ahmed
Published: November 2017

In this paper, we consider cooperative control issues for a multi-unmanned aerial vehicle (UAV) system. We propose a cooperative formation control strategy with unidirectional network connections between UAVs. Our strategy is to apply a consensus-based algorithm to the UAVs so that they can cooperatively fly in formation. First, we show that UAV models on the horizontal plane and in the vertical direction are expressed as a fourth- and second-order system, respectively. Then, we show that the stability discriminants of the multi-UAV system on the horizontal plane and in the vertical direction are expressed as polynomials. For a network structure composed of bidirectional or unidirectional network connections under the assumption that the network has a directed spanning tree, we provide conditions for formation control gains such that all roots of the polynomials have negative real parts in order for the UAVs to asymptotically converge to the positions for a desired formation by using the generalized Routh stability criterion. The proposed control algorithms are validated through simulations, and experiments are performed on multiple commercial small UAVs to validate the proposed control algorithm.
Biodiesel Production From Waste Palm Oil Using Palm Empty Fruit Bunch-Derived Novel Carbon Acid Catalyst
Authors: Thushari P. G. I. , Babel S.
Published: November 2017

Production of biodiesel from waste palm oil (WPO) can provide alternative energy and at the same time reduce the problems created by disposal of WPO. In this study, a novel, inexpensive, and environmental benign carbon acid catalyst is prepared by direct in situ concentrated H2SO4 impregnation of palm empty fruit bunch (PEFB) powder and employed for biodiesel production using WPO. The structure and the physiochemical properties of the prepared catalyst (PEFB-DS-SO3H) are analyzed by acid-base back titration data, energy dispersive X-ray spectroscopy (scanning electron microscopy (SEM)-EDS), SEM, Fourier transform infrared (FT-IR) spectroscopy, X-ray photoelectron spectroscopy (XPS), and using N2 adsorption and desorption isotherm data. It is observed that the catalyst has a unique amorphous structure with total acid density of 5.40 mmolg−1, surface area of 5.5 m2g−1, and 0.31 cm3g−1 pore volume. In addition, FT-IR, XPS, and EDS results confirm a successful sulfonation during the catalyst preparation. It is found that fatty acid methyl ester (FAME) yield increases with increasing methanol:oil (molar ratio) and reaction time up to an optimum value. The highest biodiesel yield of 91% is reported under reaction conditions of 5 wt % catalyst, 14:1 methanol: oil (molar ratio), at 65–70 °C after 14 h in an open reflux system. Results show that the catalyst can be reused for four consecutive cycles without significant loss of catalytic activity. Fuel properties of the produced biodiesel are compatible with the international fuel standards for biodiesel.
Cost-Optimal Coordination of Interacting HVAC Loads in Buildings
Authors: Bashash Saeid
Published: November 2017

This paper investigates the optimal coordination of multiple interacting heating, ventilation, and air conditioning (HVAC) appliances in buildings such as air conditioners and refrigerators, in time-varying electricity pricing environments. Each load is modeled as a first-order differential equation with a binary (ON–OFF) switching control function. An energy cost minimization problem is then formulated with weighted penalties on the temperature deviation from the desired setpoint and the control input fluctuation. Using the dynamic programming (DP) method, the cost-optimal trajectories are computed, which indicate precooling of the loads in anticipation of higher electricity prices. Moreover, the loads are desynchronized in the presence of local renewable generation to maximize the on-site consumption of the local energy. The presented results provide useful insights for the development of predictive control policies for optimal energy management in buildings.
A Further Study on the Analytical Model for the Permeability in Flip-Chip Packaging
Authors: Yao X. J. , Fang J. J. , Zhang Wenjun
Published: November 2017

The notion of permeability is very important in understanding and modeling the flow behavior of fluids in a special type of porous medium (i.e., the underfill flow in flip-chip packaging). This paper presents a new concept regarding permeability in a porous medium, namely two types of permeability: superficial permeability (with consideration of both the pore cross-sectional area and the solid matrix cross-sectional area) and pore permeability (with consideration of the pore cross-sectional area only). Subsequently, the paper proposes an analytical model (i.e., equation) for the pore permeability and superficial permeability of an underfill porous medium in a flip-chip packaging, respectively. The proposed model along with several similar models in literature is compared with a reliable numerical model developed with the computational fluid dynamics (CFD) technique, and the result of the comparison shows that the proposed model for permeability is the most accurate one among all the analytical models in literature. The main contributions of the paper are as follows: (1) the provision of a more accurate analytical model for the permeability of an underfill porous medium in flip-chip packaging, (2) the finding of two types of permeability depending on how the cross-sectional area is taken, and (3) the correction of an error in the others' model in literature.
Effect of Cr and Thermomechanical Processing on the Microstructure and Mechanical Properties of Advanced High-Strength Steel
Authors: El-Morsy Abdel-Wahab , Farahat Ahmed I. Z.
Published: November 2017

In this work, two advanced high-strength steels (AHSS) have been developed by designing alloy systems with suitable alloying elements, Mn, Si, Al, and Cr, and postforming heat treatment processes. Thermomechanical process of ∼90% forging reductions has been applied on the designed alloys at a temperature of 1100 °C, followed by austenitizing above AC3. Four cooling rates, air-cooling, air-cooling with tempering, oil quenching with tempering, and water quenching with tempering, have been applied on the forged samples. The results revealed that the estimated tensile properties of the ferrite/bainite microstructures of alloy A, without Cr, is situated between the bands of the first and the current third generation AHSS, whereas the estimated properties corresponding to the ferrite/fine bainite with 8% retained austenite of alloy B, with Cr, is overlapped with the properties exhibited by the current third generation of AHSSs. The thermomechanical process conducted on the alloy containing Cr has developed steel with tensile strength up to 1790 MPa.
Accumulation of 5-oxoproline in myocardial dysfunction and the protective effects of OPLAH
Authors: van der Pol Atze , Gil Andres , Silljé Herman H. W. , Tromp Jasper , Ovchinnikova Ekaterina S. , Vreeswijk-Baudoin Inge , Hoes Martijn , Domian Ibrahim J. , van de Sluis Bart , van Deursen Jan M. , Voors Adriaan A. , van Veldhuisen Dirk J. , van Gilst Wiek H. , Berezikov Eugene , van der Harst Pim , de Boer Rudolf A. , Bischoff Rainer , van der Meer Peter
Published: November 2017

OPLAH is cardioprotective for myocardial infarction, and its substrate 5-oxoproline is a putative circulating marker for heart failure.
Stress hormones promote EGFR inhibitor resistance in NSCLC: Implications for combinations with β-blockers
Authors: Nilsson Monique B. , Sun Huiying , Diao Lixia , Tong Pan , Liu Diane , Li Lerong , Fan Youhong , Poteete Alissa , Lim Seung-Oe , Howells Kathryn , Haddad Vincent , Gomez Daniel , Tran Hai , Pena Guillermo Armaiz , Sequist Lecia V. , Yang James C. , Wang Jing , Kim Edward S. , Herbst Roy S. , Lee J. Jack , Hong Waun Ki , Wistuba Ignacio , Hung Mien-Chie , Sood Anil K. , Heymach John V.
Published: November 2017

Chronic stress hormones promote EGFR inhibitor resistance in EGFR mutant non–small cell lung cancer.
Getting in touch with your inner stomach
Authors: Jewell Christopher M.
Published: November 2017

An ingestible and mechanically flexible piezoelectric device enables sensing of changes in the GI tract of pigs.
Predicting Invasive Aspergillosis in Hematology Patients by Combining Clinical and Genetic Risk Factors with Early Diagnostic Biomarkers
Authors: White P. Lewis , Parr Christian , Barnes Rosemary A.
Published: November 2017

Personalized medicine provides a strategic approach to the management of IA. The incidence of IA in high-risk hematology populations is relatively low (<10%), despite unavoidable
Aspergillus
exposure in patients with a potentially similar clinical risk. Nonclinical variables, including genetic mutations that increase susceptibility to IA, could explain why only certain patients develop the disease. This study screened for mutations in 322 hematology patients classified according to IA status and developed a predictive model based on genetic risk, established clinical risk factors, and diagnostic biomarkers. Genetic markers were determined by real-time PCR and, with clinical risk factors and
Aspergillus
PCR results, subjected to multilogistic regression analysis to identify a best-fit model for predicting IA. The probability of IA was calculated, and an optimal threshold was determined. Mutations in dectin-1 (rs7309123) and DC-SIGN (rs11465384 and rs7248637), allogeneic stem cell transplantation, respiratory virus infection, and
Aspergillus
PCR positivity were all significant risk factors for developing IA and were combined in a predictive model. An optimal threshold requiring three positive factors generated a mean sensitivity/specificity of 70.4%/89.2% and a probability of developing IA of 56.7%. In patients with no risk factors, the probability of developing IA was 2.4%, compared to >79.1% in patients with four or more factors. Using a risk threshold of 50%, preemptive therapy would have been prescribed for 8.4% of the population. This pilot study shows that patients can be stratified according to risk of IA, providing personalized medicine based on strategic evidence for the management of IA. Further studies are required to confirm this approach.
Use of the Accelerate Pheno System for Identification and Antimicrobial Susceptibility Testing of Pathogens in Positive Blood Cultures and Impact on Time to Results and Workflow
Authors: Charnot-Katsikas Angella , Tesic Vera , Love Nedra , Hill Brandy , Bethel Cindy , Boonlayangoor Sue , Beavis Kathleen G.
Published: November 2017

The Accelerate Pheno system uses automated fluorescence
in situ
hybridization technology with morphokinetic cellular analysis to provide rapid species identification (ID) and antimicrobial susceptibility testing (AST) results for the most commonly identified organisms in bloodstream infections. The objective was to evaluate the accuracy and workflow of bacterial and yeast ID and bacterial AST using the Accelerate Pheno system in the clinical microbiology laboratory. The consecutive fresh blood cultures received in the laboratory were analyzed by the Accelerate Pheno system within 0 to 8 h of growth detection. ID/AST performance, the average times to results, and workflow were compared to those of the routine standard of care. Of the 232 blood cultures evaluated (223 monomicrobial and 9 polymicrobial) comprising 241 organisms, the overall sensitivity and specificity for the identification of organisms were 95.6% and 99.5%, respectively. For antimicrobial susceptibility, the overall essential agreement was 95.1% and categorical agreement was 95.5% compared to routine methods. There was one very major error and 3 major errors. The time to identification and the time to susceptibility using the Accelerate Pheno system were decreased by 23.47 and 41.86 h, respectively, compared to those for the standard of care. The reduction in hands on time was 25.5 min per culture. The Accelerate Pheno system provides rapid and accurate ID/AST results for most of the organisms found routinely in blood cultures. It is easy to use, reduces hands on time for ID/AST of common blood pathogens, and enables clinically actionable results to be released much earlier than with the current standard of care.
Highly Sensitive Detection of Isoniazid Heteroresistance in Mycobacterium tuberculosis by DeepMelt Assay
Authors: Liang Bin , Tan Yaoju , Li Zi , Tian Xueshan , Du Chen , Li Hui , Li Guoli , Yao Xiangyang , Wang Zhongan , Xu Ye , Li Qingge
Published: November 2017

Detection of heteroresistance of
Mycobacterium tuberculosis
remains challenging using current genotypic drug susceptibility testing methods. Here, we described a melting curve analysis-based approach, termed DeepMelt, that can detect less-abundant mutants through selective clamping of the wild type in mixed populations. The singleplex DeepMelt assay detected 0.01%
katG
S315T in 10
5
M. tuberculosis
genomes/μl. The multiplex DeepMelt TB/INH detected 1% of mutant species in the four loci associated with isoniazid resistance in 10
4
M. tuberculosis
genomes/μl. The DeepMelt TB/INH assay was tested on a panel of DNA extracted from 602 precharacterized clinical isolates. Using the 1% proportion method as the gold standard, the sensitivity was found to be increased from 93.6% (176/188, 95% confidence interval [CI] = 89.2 to 96.3%) to 95.7% (180/188, 95% CI = 91.8 to 97.8%) compared to the MeltPro TB/INH assay. Further evaluation of 109 smear-positive sputum specimens increased the sensitivity from 83.3% (20/24, 95% CI = 64.2 to 93.3%) to 91.7% (22/24, 95% CI = 74.2 to 97.7%). In both cases, the specificity remained nearly unchanged. All heteroresistant samples newly identified by the DeepMelt TB/INH assay were confirmed by DNA sequencing and even partially by digital PCR. The DeepMelt assay may fill the gap between current genotypic and phenotypic drug susceptibility testing for detecting drug-resistant tuberculosis patients.
Multicenter Evaluation of the Modified Carbapenem Inactivation Method and the Carba NP for Detection of Carbapenemase-Producing Pseudomonas aeruginosa and Acinetobacter baumannii
Authors: Simner Patricia J. , Johnson J. Kristie , Brasso William B. , Anderson Karen , Lonsway David R. , Pierce Virginia M. , Bobenchik April M. , Lockett Zabrina C. , Charnot-Katsikas Angella , Westblade Lars F. , Yoo Brian B. , Jenkins Stephen G. , Limbago Brandi M. , Das Sanchita , Roe-Carpenter Darcie E.
Published: November 2017

The purpose of this study was to develop the modified carbapenem inactivation method (mCIM) for the detection of carbapenemase-producing
Pseudomonas aeruginosa
(CP-PA) and carbapenemase-producing
Acinetobacter baumannii
(CP-AB) and perform a multicenter evaluation of the mCIM and Carba NP tests for these nonfermenters. Thirty
P. aeruginosa
and 30
A. baumannii
isolates previously characterized by whole-genome sequencing from the CDC-FDA Antibiotic Resistance Isolate Bank were evaluated, including CP isolates (Ambler class A, B, and D), non-carbapenemase-producing (non-CP) carbapenem-resistant isolates, and carbapenem-susceptible isolates. Initial comparison of a 1-μl versus 10-μl loop inoculum for the mCIM was performed by two testing sites and showed that 10 μl was required for reliable detection of carbapenemase production among
P. aeruginosa
and
A. baumannii
. Ten testing sites then evaluated the mCIM using a 10-μl loop inoculum. Overall, the mean sensitivity and specificity of the mCIM for detection of CP-PA across all 10 sites were 98.0% (95% confidence interval [CI], 94.3 to 99.6; range, 86.7 to 100) and 95% (95% CI, 89.8 to 97.7; range, 93.3 to 100), whereas the mean sensitivity and specificity among CP-AB were 79.8% (95% CI, 74.0 to 84.9; range, 36.3 to 95.7) and 52.9% (95% CI, 40.6 to 64.9; range, 28.6 to 100), respectively. At three sites that evaluated the performance of the Carba NP test using the same set of isolates, the mean sensitivity and specificity of the Carba NP test were 97.8% (95% CI, 88.2 to 99.9; range, 93.3 to 100) and 97.8% (95% CI, 88.2 to 99.9; range, 93.3 to 100) for
P. aeruginosa
and 18.8% (95% CI, 10.4 to 30.1; range, 8.7 to 26.1) and 100% (95% CI, 83.9 to 100; range, 100) for
A. baumannii
. Overall, we found both the mCIM and the Carba NP test to be accurate for detection of carbapenemase production among
P. aeruginosa
isolates and less reliable for use with
A. baumannii
isolates.
Multicenter Evaluation of the Xpert MRSA NxG Assay for Detection of Methicillin-Resistant Staphylococcus aureus in Nasal Swabs
Authors: Yarbrough Melanie L. , Warren David K. , Allen Karen , Burkholder Dennis , Daum Robert , Donskey Curtis , Knaack Dennis , LaMarca Anthony , May Larissa , Miller Loren G. , Parenti David M. , Peterson Lance , Tan Thean Yen , Widen Raymond , Hernandez Diana R. , Wolk Donna M. , Burnham Carey-Ann D.
Published: November 2017

Health care-associated methicillin-resistant
Staphylococcus aureus
(MRSA) infections are a burden on the health care system. Clinical laboratories play a key role in reducing this burden, as the timely identification of MRSA colonization or infection facilitates infection control practices that are effective at limiting invasive MRSA infections. The Xpert MRSA NxG assay recently received FDA clearance for the direct detection of MRSA from nasal swabs. This multicenter study evaluated the clinical performance characteristics of the Xpert MRSA NxG assay with prospectively collected rayon nasal swabs (
n
= 1,103) and flocked swab (ESwab) nasal specimens (
n
= 846). Culture-based identification methods and antimicrobial susceptibility testing were used as the reference standards for comparison. According to the reference method, the positivity rates for MRSA in the population evaluated were 11.1% (122/1,103) for rayon swabs and 11.6% (98/846) for flocked swabs. The overall sensitivity and specificity of the rayon swabs were 91.0% (95% confidence interval [CI], 84.6 to 94.9%) and 96.9% (95% CI, 95.7 to 97.8%), respectively, across eight testing sites. The flocked swab specimens were 92.9% sensitive (95% CI, 86.0 to 96.5%) and 97.6% specific (95% CI, 96.2 to 98.5%) for MRSA detection across six testing sites. The sensitivity and specificity of the combined flocked and rayon swab data were 91.8% (95% CI, 87.4 to 94.8%) and 97.2% (95% CI, 96.3 to 97.9%), respectively. The positive predictive value (PPV) for rayon swabs was 78.7%, versus 83.5% for ESwabs. The negative predictive values (NPVs) for rayon swabs and ESwab specimens were 98.9% and 99.1%, respectively. In conclusion, the Xpert MRSA NxG assay is a sensitive and specific assay for the direct detection of MRSA from nasal swab specimens.
Evaluation of In-House and Commercial Serological Tests for Diagnosis of Human Tularemia
Authors: Yanes Hadjila , Hennebique Aurélie , Pelloux Isabelle , Boisset Sandrine , Bicout Dominique J. , Caspar Yvan , Maurin Max
Published: November 2017

Tularemia is a zoonosis caused by the bacterium
Francisella tularensis
. Its specific diagnosis remains based on serological methods, while
F. tularensis
is rarely detected in clinical samples by culture or PCR. The aim of the present study was to evaluate the performance of the Serion enzyme-linked immunosorbent assay (ELISA) classic
Francisella tularensis
IgG and IgM tests (Virion/Serion GmbH Institute, Würzburg, Germany) and the VIRapid tularemia immunochromatographic test (ICT) (Vircell, Granada, Spain) compared to that of the in-house microagglutination test (MAT) and indirect immunofluorescence assay (IFA) currently used at the French National Reference Center for
Francisella
. We evaluated 256 consecutive sera from 208 patients, including 51 confirmed and 23 probable tularemia cases, and 134 control patients not infected with
F. tularensis
. The IFA tests displayed 72.5% sensitivity for IgM (cutoff titer ≥80) and 74.5% for IgG (cutoff titer ≥160), and 99.3% specificity for both IgM and IgG. Using cutoffs advocated by the manufacturer, the Serion ELISAs displayed 88.2% sensitivity for IgM and 86.3% for IgG antibodies; specificity was 94.8% for IgM and 95.5% for IgG. Compared to MAT and IFA tests, the Serion ELISAs allowed earlier detection of specific antibodies (1 to 2 weeks versus 2 to 3 weeks after the onset of symptoms). The ICT sensitivity and specificity were 90% and 83.6%, respectively, when considering the cutoff advocated by the manufacturer. In conclusion, the Serion ELISAs are useful as screening tests for tularemia diagnosis, but additional confirmatory tests (such as MAT and IFA) are needed, especially in areas of low endemicity.
Unique Directional Motility of Influenza C Virus Controlled by Its Filamentous Morphology and Short-Range Motions
Authors: Sakai Tatsuya , Takagi Hiroaki , Muraki Yasushi , Saito Mineki
Published: November 2017

Influenza virus motility is based on cooperation between two viral spike proteins, hemagglutinin (HA) and neuraminidase (NA), and is a major determinant of virus infectivity. To translocate a virus particle on the cell surface, HA molecules exchange viral receptors and NA molecules accelerate the receptor exchange of HA. This type of virus motility was recently identified in influenza A virus (IAV). To determine if other influenza virus types have a similar receptor exchange mechanism-driven motility, we investigated influenza C virus (ICV) motility on a receptor-fixed glass surface. This system excludes receptor mobility, which makes it more desirable than a cell surface for demonstrating virus motility by receptor exchange. Like IAV, ICV was observed to move across the receptor-fixed surface. However, in contrast to the random movement of IAV, a filamentous ICV strain, Ann Arbor/1/50 (AA), moved in a straight line, in a directed manner, and at a constant rate, whereas a spherical ICV strain, Taylor/1233/47 (Taylor), moved randomly, similar to IAV. The AA and Taylor viruses each moved with a combination of gradual (crawling) and rapid (gliding) motions, but the distances of crawling and gliding for the AA virus were shorter than those of the Taylor virus. Our findings indicate that like IAV, ICV also has a motility that is driven by the receptor exchange mechanism. However, compared with IAV movement, filamentous ICV movement is highly regulated in both direction and speed. Control of ICV movement is based on its specific motility employing short crawling and gliding motions as well as its own filamentous morphology.

IMPORTANCE
Influenza virus enters into a host cell for infection via cellular endocytosis. Human influenza virus infects epithelial cells of the respiratory tract, the surfaces of which are hidden by abundant cilia that are inactive in endocytosis. An open question is the manner by which the virus migrates to endocytosis-active domains. In analyzing individual virus behaviors through single-virus tracking, we identified a novel function of the hemagglutinin and esterase of influenza C virus (ICV) as the motility machinery. Hemagglutinin iteratively exchanges a viral receptor, causing virus movement. Esterase degrades the receptors along the trajectory traveled by the virus and prevents the virus from moving backward, causing directional movement. We propose that ICV has a unique motile machinery directionally controlled via hemagglutinin sensing the receptor density manipulated by esterase.
A Y161F Hemagglutinin Substitution Increases Thermostability and Improves Yields of 2009 H1N1 Influenza A Virus in Cells
Authors: Wen Feng , Li Lei , Zhao Nan , Chiang Meng-Jung , Xie Hang , Cooley Jim , Webby Richard , Wang Peng George , Wan Xiu-Feng
Published: November 2017

Vaccination is the primary strategy for influenza prevention and control. However, egg-based vaccines, the predominant production platform, have several disadvantages, including the emergence of viral antigenic variants that can be induced during egg passage. These limitations have prompted the development of cell-based vaccines, which themselves are not without issue. Most importantly, vaccine seed viruses often do not grow efficiently in mammalian cell lines. Here we aimed to identify novel high-yield signatures for influenza viruses in continuous Madin-Darby canine kidney (MDCK) and Vero cells. Using influenza A(H1N1)pdm09 virus as the testing platform and an integrating error-prone PCR-based mutagenesis strategy, we identified a Y161F mutation in hemagglutinin (HA) that not only enhanced the infectivity of the resultant virus by more than 300-fold but also increased its thermostability without changing its original antigenic properties. The vaccine produced from the Y161F mutant fully protected mice against lethal challenge with wild-type A(H1N1)pdm09. Compared with A(H1N1)pdm09, the Y161F mutant had significantly higher avidity for avian-like and human-like receptor analogs. Of note, the introduction of the Y161F mutation into HA of seasonal H3N2 influenza A virus (IAV) and canine H3N8 IAV also increased yields and thermostability in MDCK cells and chicken embryotic eggs. Thus, residue F161 plays an important role in determining viral growth and thermostability, which could be harnessed to optimize IAV vaccine seed viruses.

IMPORTANCE
Although a promising complement to current egg-based influenza vaccines, cell-based vaccines have one large challenge: high-yield vaccine seeds for production. In this study, we identified a molecular signature, Y161F, in hemagglutinin (HA) that resulted in increased virus growth in Madin-Darby canine kidney and Vero cells, two cell lines commonly used for influenza vaccine manufacturing. This Y161F mutation not only increased HA thermostability but also enhanced its binding affinity for α2,6- and α2,3-linked Neu5Ac. These results suggest that a vaccine strain bearing the Y161F mutation in HA could potentially increase vaccine yields in mammalian cell culture systems.
Estimating Initial Viral Levels during Simian Immunodeficiency Virus/Human Immunodeficiency Virus Reactivation from Latency
Authors: Pinkevych Mykola , Fennessey Christine M. , Cromer Deborah , Tolstrup Martin , Søgaard Ole S. , Rasmussen Thomas A. , Keele Brandon F. , Davenport Miles P.
Published: November 2017

Human immunodeficiency virus (HIV) viremia rebounds rapidly after treatment interruption, and a variety of strategies are being explored to reduce or control viral reactivation posttreatment. This viral rebound arises from reactivation of individual latently infected cells, which spread during ongoing rounds of productive infection. The level of virus produced by the initial individual reactivating cells is not known, although it may have major implications for the ability of different immune interventions to control viral rebound. Here we use data from both HIV and simian immunodeficiency virus (SIV) treatment interruption studies to estimate the initial viral load postinterruption and thereby the initial individual reactivation event. Using a barcoded virus (SIVmac239M) to track reactivation from individual latent cells, we use the observed viral growth rates and frequency of reactivation to model the dynamics of reactivation to estimate that a single reactivated latent cell can produce an average viral load equivalent to ∼0.1 to 0.5 viral RNA (vRNA) copies/ml. Modeling of treatment interruption in HIV suggests an initial viral load equivalent of ∼0.6 to 1 vRNA copies/ml. These low viral loads immediately following latent cell reactivation provide a window of opportunity for viral control by host immunity, before further replication allows viral spread. This work shows the initial levels of viral production that must be controlled in order to successfully suppress HIV reactivation following treatment interruption.

IMPORTANCE
Current treatment for HIV is able to suppress viral replication and prevent disease progression. However, treatment cannot eradicate infection, because the virus lies silent within latently infected cells. If treatment is stopped, the virus usually rebounds above the level of detection within a few weeks. There are a number of approaches being tested aimed at either eradicating latently infected cells or controlling the virus if it returns. Studying both the small pool of latently infected cells and the early events during viral reactivation is difficult, because these involve very small levels of virus that are difficult to measure directly. Here, we combine experimental data and mathematical modeling to understand the very early events during viral reactivation from latency in both HIV infection of humans and SIV infection of monkeys. We find that the initial levels of virus are low, which may help in designing therapies to control early viral reactivation.
ALT-803 Transiently Reduces Simian Immunodeficiency Virus Replication in the Absence of Antiretroviral Treatment
Authors: Ellis-Connell Amy L. , Balgeman Alexis J. , Zarbock Katie R. , Barry Gabrielle , Weiler Andrea , Egan Jack O. , Jeng Emily K. , Friedrich Thomas , Miller Jeffrey S. , Haase Ashley T. , Schacker Timothy W. , Wong Hing C. , Rakasz Eva , O'Connor Shelby L.
Published: November 2017

Developing biological interventions to control human immunodeficiency virus (HIV) replication in the absence of antiretroviral therapy (ART) could contribute to the development of a functional cure. As a potential alternative to ART, the interleukin-15 (IL-15) superagonist ALT-803 has been shown to boost the number and function of HIV-specific CD8+T and NK cell populationsin vitro. Four simian immunodeficiency virus (SIV)-positive rhesus macaques, three of whom possessed major histocompatibility complex alleles associated with control of SIV and all of whom had received SIV vaccine vectors that had the potential to elicit CD8+T cell responses, were given ALT-803 in three treatment cycles. The first and second cycles of treatment were separated by 2 weeks, while the third cycle was administered after a 29-week break. ALT-803 transiently elevated the total CD8+effector and central memory T cell and NK cell populations in peripheral blood, while viral loads transiently decreased by ∼2 logs in all animals. Virus suppression was not sustained as T cells became less responsive to ALT-803 and waned in numbers. No effect on viral loads was observed in the second cycle of ALT-803, concurrent with downregulation of the IL-2/15 common γC and β chain receptors on both CD8+T cells and NK cells. Furthermore, populations of immunosuppressive T cells increased during the second cycle of ALT-803 treatment. During the third treatment cycle, responsiveness to ALT-803 was restored. CD8+T cells and NK cells increased again 3- to 5-fold, and viral loads transiently decreased again by 1 to 2 logs.

IMPORTANCEOverall, our data show that ALT-803 has the potential to be used as an immunomodulatory agent to elicit effective immune control of HIV/SIV replication. We identify mechanisms to explain why virus control is transient, so that this model can be used to define a clinically appropriate treatment regimen.
Analyses of the Kroon Virus Major Capsid Gene and Its Transcript Highlight a Distinct Pattern of Gene Evolution and Splicing among Mimiviruses
Authors: Boratto Paulo Victor Miranda , Dornas Fábio Pio , da Silva Lorena Christine Ferreira , Rodrigues Rodrigo Araújo Lima , Oliveira Graziele Pereira , Cortines Juliana Reis , Drumond Betânia Paiva , Abrahão Jônatas Santos
Published: November 2017

The inclusion of
Mimiviridae
members in the putative monophyletic nucleocytoplasmic large DNA virus (NCLDV) group is based on genomic and phylogenomic patterns. This shows that, along with other viral families, they share a set of genes known as core or “hallmark genes,” including the gene for the major capsid protein (MCP). Although previous studies have suggested that the maturation of mimivirus MCP transcripts is dependent on splicing, there is little information about the processing of this transcript in other mimivirus isolates. Here we report the characterization of a new mimivirus isolate, called Kroon virus (KV) mimivirus. Analysis of the structure, synteny, and phylogenetic relationships of the MCP genes in many mimivirus isolates revealed a remarkable variation at position and types of intronic and exonic regions, even for mimiviruses belonging to the same lineage. In addition, sequencing of KV and Acanthamoeba polyphaga mimivirus (APMV) MCP transcripts has shown that inside the family, even related giant viruses may present different ways to process the MCP mRNA. These results contribute to the understanding of the genetic organization and evolution of the MCP gene in mimiviruses.

IMPORTANCE
Mimivirus isolates have been obtained by prospecting studies since 2003. Based on genomic and phylogenomic studies of conserved genes, these viruses have been clustered together with members of six other viral families. Although the major capsid protein (MCP) gene is an important member of the so-called “hallmark genes,” there is little information about the processing and structure of this gene in many mimivirus isolates. In this work, we have analyzed the structure, synteny, and phylogenetic relationships of the MCP genes in many mimivirus isolates; these genes showed remarkable variation at position and types of intronic and exonic regions, even for mimiviruses belonging to the same lineage. These results contribute to the understanding of the genetic organization and evolution of the MCP gene in mimiviruses.
MCR-1 Inhibition with Peptide-Conjugated Phosphorodiamidate Morpholino Oligomers Restores Sensitivity to Polymyxin in
Escherichia coli
Authors: Daly Seth M. , Sturge Carolyn R. , Felder-Scott Christina F. , Geller Bruce L. , Greenberg David E.
Published: November 2017

In late 2015, the first example of a transferrable polymyxin resistance mechanism in Gram-negative pathogens, MCR-1, was reported. Since that report, MCR-1 has been described to occur in many Gram-negative pathogens, and the mechanism of MCR-1-mediated resistance was rapidly determined: an ethanolamine is attached to lipid A phosphate groups, rendering the membrane more electropositive and repelling positively charged polymyxins. Acquisition of MCR-1 is clinically significant because polymyxins are frequently last-line antibiotics used to treat extensively resistant organisms, so acquisition of this mechanism might lead to pan-resistant strains. Therefore, the ability to inhibit MCR-1 and restore polymyxin sensitivity would be an important scientific advancement. Peptide-conjugated phosphorodiamidate morpholino oligomers (PPMOs) are antisense molecules that were designed to target mRNA, preventing translation. Peptide conjugation enhances cellular entry, but they are positively charged, so we tested our lead antibacterial PPMOs by targeting an essential
Escherichia coli
gene,
acpP
, and demonstrated that they were still effective in
mcr
-
1
-positive
E. coli
strains. We then designed and synthesized two PPMOs targeted to
mcr-1
mRNA. Five clinical
mcr-1
-positive
E. coli
strains were resensitized to polymyxins by MCR-1 inhibition, reducing MICs 2- to 16-fold. Finally, therapeutic dosing of BALB/c mice with MCR-1 PPMO combined with colistin in a sepsis model reduced morbidity and bacterial burden in the spleen at 24 h and offered a survival advantage out to 5 days. This is the first example of a way to modulate colistin resistance with an antisense approach and may be a viable strategy to combat this globally emerging antibiotic resistance threat.

IMPORTANCE
Polymyxin use has been increasing as a last line of defense against Gram-negative pathogens with high-level resistance mechanisms, such as carbapenemases. The recently described MCR-1 is a plasmid-mediated mechanism of resistance to polymyxins. MCR-1 is currently found in Gram-negative organisms already possessing high-level resistance mechanisms, leaving clinicians few or no antibacterial options for infections caused by these strains. This study utilizes antisense molecules that target mRNA, preventing protein translation. Herein we describe antisense molecules that can be directly antibacterial because they target genes essential to bacterial growth or blockade of MCR-1, restoring polymyxin sensitivity. We also demonstrate that MCR-1 antisense molecules restore the efficacies of polymyxins in mouse models of
E. coli
septicemia. Considering all things together, we demonstrate that antisense molecules may be effective therapeutics either alone when they target an essential gene or combined with antibiotics when they target specific resistance mechanisms, such as those seen with MCR-1.
The First Microbial Colonizers of the Human Gut: Composition, Activities, and Health Implications of the Infant Gut Microbiota
Authors: Milani Christian , Duranti Sabrina , Bottacini Francesca , Casey Eoghan , Turroni Francesca , Mahony Jennifer , Belzer Clara , Delgado Palacio Susana , Arboleya Montes Silvia , Mancabelli Leonardo , Lugli Gabriele Andrea , Rodriguez Juan Miguel , Bode Lars , de Vos Willem , Gueimonde Miguel , Margolles Abelardo , van Sinderen Douwe , Ventura Marco
Published: November 2017

The human gut microbiota is engaged in multiple interactions affecting host health during the host's entire life span. Microbes colonize the neonatal gut immediately following birth. The establishment and interactive development of this early gut microbiota are believed to be (at least partially) driven and modulated by specific compounds present in human milk. It has been shown that certain genomes of infant gut commensals, in particular those of bifidobacterial species, are genetically adapted to utilize specific glycans of this human secretory fluid, thus representing a very intriguing example of host-microbe coevolution, where both partners are believed to benefit. In recent years, various metagenomic studies have tried to dissect the composition and functionality of the infant gut microbiome and to explore the distribution across the different ecological niches of the infant gut biogeography of the corresponding microbial consortia, including those corresponding to bacteria and viruses, in healthy and ill subjects. Such analyses have linked certain features of the microbiota/microbiome, such as reduced diversity or aberrant composition, to intestinal illnesses in infants or disease states that are manifested at later stages of life, including asthma, inflammatory bowel disease, and metabolic disorders. Thus, a growing number of studies have reported on how the early human gut microbiota composition/development may affect risk factors related to adult health conditions. This concept has fueled the development of strategies to shape the infant microbiota composition based on various functional food products. In this review, we describe the infant microbiota, the mechanisms that drive its establishment and composition, and how microbial consortia may be molded by natural or artificial interventions. Finally, we discuss the relevance of key microbial players of the infant gut microbiota, in particular bifidobacteria, with respect to their role in health and disease.
Chronic inflammation is a feature of Achilles tendinopathy and rupture
Authors: Dakin Stephanie Georgina , Newton Julia , Martinez Fernando O , Hedley Robert , Gwilym Stephen , Jones Natasha , Reid Hamish A B , Wood Simon , Wells Graham , Appleton Louise , Wheway Kim , Watkins Bridget , Carr Andrew Jonathan
Published: November 2017

Background

Recent investigation of human tissue and cells from positional tendons such as the rotator cuff has clarified the importance of inflammation in the development and progression of tendon disease. These mechanisms remain poorly understood in disease of energy-storing tendons such as the Achilles. Using tissue biopsies from patients, we investigated if inflammation is a feature of Achilles tendinopathy and rupture.

Methods

We studied Achilles tendon biopsies from symptomatic patients with either mid-portion tendinopathy or rupture for evidence of abnormal inflammatory signatures. Tendon-derived stromal cells from healthy hamstring and diseased Achilles were cultured to determine the effects of cytokine treatment on expression of inflammatory markers.

Results

Tendinopathic and ruptured Achilles highly expressed CD14+ and CD68+ cells and showed a complex inflammation signature, involving NF-κB, interferon and STAT-6 activation pathways. Interferon markers IRF1 and IRF5 were highly expressed in tendinopathic samples. Achilles ruptures showed increasedPTGS2andinterleukin-8expression. Tendinopathic and ruptured Achilles tissues expressed stromal fibroblast activation markers podoplanin and CD106. Tendon cells isolated from diseased Achilles showed increased expression of pro-inflammatory and stromal fibroblast activation markers after cytokine stimulation compared with healthy hamstring tendon cells.

Conclusions

Tissue and cells derived from tendinopathic and ruptured Achilles tendons show evidence of chronic (non-resolving) inflammation. The energy-storing Achilles shares common cellular and molecular inflammatory mechanisms with functionally distinct rotator cuff positional tendons. Differences seen in the profile of ruptured Achilles are likely to be attributable to a superimposed phase of acute inflammation and neo-vascularisation. Strategies that target chronic inflammation are of potential therapeutic benefit for patients with Achilles tendon disease.
Vitamin D status in chronic fatigue syndrome/myalgic encephalomyelitis: a cohort study from the North-West of England
Authors: Earl Kate E , Sakellariou Giorgos K , Sinclair Melanie , Fenech Manuel , Croden Fiona , Owens Daniel J , Tang Jonathan , Miller Alastair , Lawton Clare , Dye Louise , Close Graeme L , Fraser William D , McArdle Anne , Beadsworth Michael B J
Published: November 2017

Objective

Severe vitamin D deficiency is a recognised cause of skeletal muscle fatigue and myopathy. The aim of this study was to examine whether chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) is associated with altered circulating vitamin D metabolites.

Design

Cohort study.

Setting

UK university hospital, recruiting from April 2014 to April 2015.

Participants

Ninety-two patients with CFS/ME and 94 age-matched healthy controls (HCs).

Main outcome measures

The presence of a significant association between CFS/ME, fatigue and vitamin D measures.

Results

No evidence of a deficiency in serum total 25(OH) vitamin D (25(OH)D2and 25(OH)D3metabolites) was evident in individuals with CFS/ME. Liquid chromatography tandem mass spectrometry (LC–MS/MS) analysis revealed that total 25(OH)D was significantly higher (p=0.001) in serum of patients with CFS/ME compared with HCs (60.2 and 47.3 nmol/L, respectively). Analysis of food/supplement diaries with WinDiets revealed that the higher total 25(OH) vitamin D concentrations observed in the CFS/ME group were associated with increased vitamin D intake through use of supplements compared with the control group. Analysis of Chalder Fatigue Questionnaire data revealed no association between perceived fatigue and vitamin D levels.

Conclusions

Low serum concentrations of total 25(OH)D do not appear to be a contributing factor to the level of fatigue of CFS/ME.
Efficacy of platelet-rich plasma injections for symptomatic tendinopathy: systematic review and meta-analysis of randomised injection-controlled trials
Authors: Miller Larry E , Parrish William R , Roides Breana , Bhattacharyya Samir
Published: November 2017

Aim

To determine the efficacy of platelet-rich plasma (PRP) injections for symptomatic tendinopathy.

Design

Systematic review of randomised, injection-controlled trials with meta-analysis.

Data sources

Systematic searches of MEDLINE and EMBASE, supplemented by manual searches.

Eligibility criteria for selecting studies

Randomised controlled trials with 3 months minimum follow-up that evaluated pain reduction with PRP versus control (saline, local anaesthetic, corticosteroid) injections in patients with symptomatic tendinopathy.

Results

A total of 16 randomised controlled trials (18 groups) of PRP versus control were included. Median sample size was 35 patients, a study size that would require an effect size ≥1.0 to achieve statistical significance. PRP was more efficacious than control in reducing tendinopathy pain, with an effect size of 0.47 (95% CI 0.22 to 0.72, p<0.001), signifying a moderate treatment effect. Heterogeneity among studies was moderate (I2=67%, p<0.001). In subgroup analysis and meta-regression, studies with a higher proportion of female patients were associated with greater treatment benefits with PRP.

Conclusions

Injection of PRP is more efficacious than control injections in patients with symptomatic tendinopathy.