bluuebunny/crossref_metadata_2025 huggingface dataset:
Predatory fish depletion and recovery potential on Caribbean reefs
Authors: Valdivia Abel , Cox Courtney Ellen , Bruno John Francis
Published: March 2017

Overfishing has eliminated up to 90% of predatory reef fishes, but recovery potential is still high in several Caribbean reefs.
A three-degree horizon of peace in the military alliance network
Authors: Li Weihua , Bradshaw Aisha E. , Clary Caitlin B. , Cranmer Skyler J.
Published: March 2017

Indirect military alliances suppress conflict between states that are up to three degrees removed in the alliance network.
Can deep reefs rescue shallow ones?
Authors: Weiss Kenneth R.
Published: March 2017

Scientists test the idea that species squeezed out of shallow coral reefs may find a refuge in deeper habitats.
Precipitation drives global variation in natural selection
Authors: Siepielski Adam M. , Morrissey Michael B. , Buoro Mathieu , Carlson Stephanie M. , Caruso Christina M. , Clegg Sonya M. , Coulson Tim , DiBattista Joseph , Gotanda Kiyoko M. , Francis Clinton D. , Hereford Joe , Kingsolver Joel G. , Augustine Kate E. , Kruuk Loeske E. B. , Martin Ryan A. , Sheldon Ben C. , Sletvold Nina , Svensson Erik I. , Wade Michael J. , MacColl Andrew D. C.
Published: March 2017

Climate-driven selection

Climate change will fundamentally alter many aspects of the natural world. To understand how species may adapt to this change, we must understand which aspects of the changing climate exert the most powerful selective forces. Siepielskiet al.looked at studies of selection across species and regions and found that, across biomes, the strongest sources of selection were precipitation and transpiration changes. Importantly, local and regional climate change explained patterns of selection much more than did global change.

Science, this issue p.959
A nontoxic pain killer designed by modeling of pathological receptor conformations
Authors: Spahn V. , Del Vecchio G. , Labuz D. , Rodriguez-Gaztelumendi A. , Massaly N. , Temp J. , Durmaz V. , Sabri P. , Reidelbach M. , Machelska H. , Weber M. , Stein C.
Published: March 2017

A pain killer without side effects

Opioids are very strong and effective pain killers. However, they also have a range of well-known side effects and can cause addiction. Painful conditions such as inflammation or trauma are often associated with localized tissue acidification. Spahn
et al.
designed a novel opioid receptor agonist that, unlike clinically used opioids, best activates the receptors in such acidified tissues. In rat models of inflammatory pain, the new drug exerted strong pain relief essentially without the side effects of standard opioids.

Science
, this issue p.
966
The atom, the molecule, and the covalent organic framework
Authors: Diercks Christian S. , Yaghi Omar M.
Published: March 2017

A framework for molecular assembly

Covalent molecular frameworks are crystalline microporous materials assembled from organic molecules through strong covalent bonds in a process termed reticular synthesis. Diercks and Yaghi review developments in this area, noting the parallels between framework assembly and the covalent assembly of atoms into molecules, as described just over a century ago by Lewis. Emerging challenges include functionalization of existing frameworks and the creation of flexible materials through the design of woven structures.

Science
, this issue p.
eaal1585
Membrane proteins scrambling through a foldinglandscape
Authors: Müller Daniel J. , Gaub Hermann E.
Published: March 2017

Improved force microscopy reveals unfolding steps with microsecond time
resolution
Durable resistance to rice blast
Authors: Wang Guo-Liang , Valent Barbara
Published: March 2017

A resistance gene pair balances immunity against crop yield
Utility of Second-Generation Line Probe Assay (Hain MTBDR
plus
) Directly on 2-Month Sputum Specimens for Monitoring Tuberculosis Treatment Response
Authors: Rockwood Neesha , Wojno Justyna , Ghebrekristos Yonas , Nicol Mark P. , Meintjes Graeme , Wilkinson Robert J.
Published: March 2017

The utility of a line probe assay (Genotype MTBDR
plus
) performed directly on 2-month sputa to monitor tuberculosis treatment response is unknown. We assessed if direct testing of 2-month sputa with MTBDR
plus
can predict 2-month culture conversion and long-term treatment outcome. Xpert MTB/RIF-confirmed rifampin-susceptible tuberculosis cases were recruited at tuberculosis diagnosis and followed up at 2 and 5 to 6 months. MTBDR
plus
was performed directly on 2-month sputa and on all positive cultured isolates at 2 and 5 to 6 months. We also investigated the association of a positive direct MTBDR
plus
at 2 months with subsequent unsuccessful tuberculosis treatment outcome (failure/death during treatment or subsequent disease recurrence). A total of 279 patients (62% of whom were HIV-1 coinfected) were recruited. Direct MTBDR
plus
at 2 months had a sensitivity of 78% (95% confidence interval [CI], 65 to 87) and specificity of 80% (95% CI, 74 to 84) to predict culture positivity at 2 months with a high negative predictive value of 93% (95% CI, 89 to 96). Inconclusive genotypic susceptibility results for both rifampin and isoniazid were seen in 26% of MTBDR
plus
tests performed directly on sputum. Compared to a reference of MTBDR
plus
performed on positive cultures, the false-positive resistance rate for direct testing of MTBDR
plus
on sputa was 4% for rifampin and 2% for isoniazid. While a positive 2-month smear was not significantly associated with an unsuccessful treatment outcome (adjusted odds ratio [aOR], 2.69; 95% CI, 0.88 to 8.21), a positive direct MTBDR
plus
at 2 months was associated with an unsuccessful outcome (aOR 2.87; 95% CI, 1.11 to 7.42). There is moderate utility of direct 2-month MTBDR
plus
to predict culture conversion at 2 months and also to predict an unfavorable outcome.
Evaluating Novel Diagnostics in an Outbreak Setting: Lessons Learned from Ebola
Authors: Pollock Nira R. , Wonderly Betsy
Published: March 2017

Inadequate access to rapid testing for Ebola virus disease during the 2014-to-2016 outbreak led to an explosion in the development of diagnostics that could be performed at or near the point of care and by less-experienced operators, leading in turn to an acute need for novel test evaluation. Here, we present the challenges to development and evaluation of novel diagnostics in an emergency setting and suggestions for potential new “global emergency standards” to address them.
A Strategy for Cultivation of Retargeted Oncolytic Herpes Simplex Viruses in Non-cancer Cells
Authors: Leoni Valerio , Gatta Valentina , Casiraghi Costanza , Nicosia Alfredo , Petrovic Biljana , Campadelli-Fiume Gabriella
Published: March 2017

The oncolytic herpes simplex virus (HSV) that has been approved for clinical practice and those HSVs in clinical trials are attenuated viruses, often with the neurovirulence gene γ
1
34.5 and additional genes deleted. One strategy to engineer nonattenuated oncolytic HSVs consists of retargeting the viral tropism to a cancer-specific receptor of choice, exemplified by HER2 (human epidermal growth factor receptor 2), which is present in breast, ovary, and other cancers, and in detargeting from the natural receptors. Because the HER2-retargeted HSVs strictly depend on this receptor for infection, the viruses employed in preclinical studies were cultivated in HER2-positive cancer cells. The production of clinical-grade viruses destined for humans should avoid the use of cancer cells. Here, we engineered the R-213 recombinant, by insertion of a 20-amino-acid (aa) short peptide (named GCN4) in the gH of R-LM113; this recombinant was retargeted to HER2 through insertion in gD of a single-chain antibody (scFv) to HER2. Next, we generated a Vero cell line expressing an artificial receptor (GCN4R) whose N terminus consists of an scFv to GCN4 and therefore is capable of interacting with GCN4 present in gH of R-213. R-213 replicated as well as R-LM113 in SK-OV-3 cells, implying that addition of the GCN4 peptide was not detrimental to gH. R-213 grew to relatively high titers in Vero-GCN4R cells, efficiently spread from cell to cell, and killed both Vero-GCN4R and SK-OV-3 cells, as expected for an oncolytic virus. Altogether, Vero-GCN4R cells represent an efficient system for cultivation of retargeted oncolytic HSVs in non-cancer cells.

IMPORTANCE
There is growing interest in viruses as oncolytic agents, which can be administered in combination with immunotherapeutic compounds, including immune checkpoint inhibitors. The oncolytic HSV approved for clinical practice and those in clinical trials are attenuated viruses. An alternative to attenuation is a cancer specificity achieved by tropism retargeting to selected cancer receptors. However, the retargeted oncolytic HSVs strictly depend on cancer receptors for infection. Here, we devised a strategy for
in vitro
cultivation of retargeted HSVs in non-cancer cells. The strategy envisions a double-retargeting approach: one retargeting is via gD to the cancer receptor, and the second retargeting is via gH to an artificial receptor expressed in Vero cells. The double-retargeted HSV uses alternatively the two receptors to infect cancer cells or producer cells. A universal non-cancer cell line for growth of clinical-grade retargeted HSVs represents a step forward in the translational phase.
Pathways of Prion Spread during Early Chronic Wasting Disease in Deer
Authors: Hoover Clare E. , Davenport Kristen A. , Henderson Davin M. , Denkers Nathaniel D. , Mathiason Candace K. , Soto Claudio , Zabel Mark D. , Hoover Edward A.
Published: March 2017

Among prion infections, two scenarios of prion spread are generally observed: (i) early lymphoid tissue replication or (ii) direct neuroinvasion without substantial antecedent lymphoid amplification. In nature, cervids are infected with chronic wasting disease (CWD) prions by oral and nasal mucosal exposure, and studies of early CWD pathogenesis have implicated pharyngeal lymphoid tissue as the earliest sites of prion accumulation. However, knowledge of chronological events in prion spread during early infection remains incomplete. To investigate this knowledge gap in early CWD pathogenesis, we exposed white-tailed deer to CWD prions by mucosal routes and performed serial necropsies to assess PrP
CWD
tissue distribution by real-time quaking-induced conversion (RT-QuIC) and tyramide signal amplification immunohistochemistry (TSA-IHC). Although PrP
CWD
was not detected by either method in the initial days (1 and 3) postexposure, we observed PrP
CWD
seeding activity and follicular immunoreactivity in oropharyngeal lymphoid tissues at 1 and 2 months postexposure (MPE). At 3 MPE, PrP
CWD
replication had expanded to all systemic lymphoid tissues. By 4 MPE, the PrP
CWD
burden in all lymphoid tissues had increased and approached levels observed in terminal disease, yet there was no evidence of nervous system invasion. These results indicate the first site of CWD prion entry is in the oropharynx, and the initial phase of prion amplification occurs in the oropharyngeal lymphoid tissues followed by rapid dissemination to systemic lymphoid tissues. This lymphoid replication phase appears to precede neuroinvasion.

IMPORTANCE
Chronic wasting disease (CWD) is a universally fatal transmissible spongiform encephalopathy affecting cervids, and natural infection occurs through oral and nasal mucosal exposure to infectious prions. Terminal disease is characterized by PrP
CWD
accumulation in the brain and lymphoid tissues of affected animals. However, the initial sites of prion accumulation and pathways of prion spread during early CWD infection remain unknown. To investigate the chronological events of early prion pathogenesis, we exposed deer to CWD prions and monitored the tissue distribution of PrP
CWD
over the first 4 months of infection. We show CWD uptake occurs in the oropharynx with initial prion replication in the draining oropharyngeal lymphoid tissues, rapidly followed by dissemination to systemic lymphoid tissues without evidence of neuroinvasion. These data highlight the two phases of CWD infection: a robust prion amplification in systemic lymphoid tissues prior to neuroinvasion and establishment of a carrier state.
C-Terminal HIV-1 Transframe p6* Tetrapeptide Blocks Enhanced Gag Cleavage Incurred by Leucine Zipper Replacement of a Deleted p6* Domain
Authors: Yu Fu-Hsien , Huang Kuo-Jung , Wang Chin-Tien
Published: March 2017

HIV-1 protease (PR) functions as a homodimer mediating virus maturation following virus budding. Gag-Pol dimerization is believed to trigger embedded PR activation by promoting PR dimer formation. Early PR activation can lead to markedly reduced virus yields due to premature Gag cleavage. The p6* peptide, located between Gag and PR, is believed to ensure virus production by preventing early PR maturation. Studies aimed at finding supporting evidence for this proposal are limited due to a reading frame overlap between p6* and the p6gag budding domain. To determine if p6* affects virus production via the modulation of PR activation, we engineered multiple constructs derived from Dp6*PR (an assembly- and processing-competent construct with Pol fused at the inactivated PR C terminus). The data indicated that a p6* deletion adjacent to active PR significantly impaired virus processing. We also observed that the insertion of a leucine zipper (LZ) dimerization motif in the deleted region eliminated virus production in a PR activity-dependent manner, suggesting that the LZ insertion triggered premature PR activation by facilitating PR dimer formation. As few as four C-terminal p6* residues remaining at the p6*/PR junction were sufficient to restore virus yields, with a Gag processing profile similar to that of the wild type. Our study provides supporting evidence in a virus assembly context that the C-terminal p6* tetrapeptide plays a role in preventing premature PR maturation.

IMPORTANCE
Supporting evidence for the assumption that p6* retards PR maturation in the context of virus assembly is lacking. We found that replacing p6* with a leucine zipper peptide abolished virus assembly due to the significant enhancement of Gag cleavage. However, as few as four C-terminal p6* residues remaining in the deleted region were sufficient for significant PR release, as well as for counteracting leucine zipper-incurred premature Gag cleavage. Our data provide evidence that (i) p6* ensures virus assembly by preventing early PR activation and (ii) four C-terminal p6* residues are critical for modulating PR activation. Current PR inhibitor development efforts are aimed largely at mature PR, but there is a tendency for HIV-1 variants that are resistant to multiple protease inhibitors to emerge. Our data support the idea of modulating PR activation by targeting PR precursors as an alternative approach to controlling HIV-1/AIDS.
Interferon Gamma Prevents Infectious Entry of Human Papillomavirus 16 via an L2-Dependent Mechanism
Authors: Day Patricia M. , Thompson Cynthia D. , Lowy Douglas R. , Schiller John T.
Published: March 2017

In this study, we report that gamma interferon (IFN-γ) treatment, but not IFN-α, -β, or -λ treatment, dramatically decreased infection of human papillomavirus 16 (HPV16) pseudovirus (PsV). In a survey of 20 additional HPV and animal papillomavirus types, we found that many, but not all, PsV types were also inhibited by IFN-γ. Microscopic and biochemical analyses of HPV16 PsV determined that the antiviral effect was exerted at the level of endosomal processing of the incoming capsid and depended on the JAK2/STAT1 pathway. In contrast to infection in the absence of IFN-γ, where L1 proteolytic products are produced during endosomal capsid processing and L2/DNA complexes segregate from L1 in the late endosome and travel to the nucleus, IFN-γ treatment led to decreased L1 proteolysis and retention of L2 and the viral genome in the late endosome/lysosome. PsV sensitivity or resistance to IFN-γ treatment was mapped to the L2 protein, as determined with infectious hybrid PsV, in which the L1 protein was derived from an IFN-γ-sensitive HPV type and the L2 protein from an IFN-γ-insensitive type or vice versa.

IMPORTANCEA subset of HPV are the causative agents of many human cancers, most notably cervical cancer. This work describes the inhibition of infection of multiple HPV types, including oncogenic types, by treatment with IFN-γ, an antiviral cytokine that is released from stimulated immune cells. Exposure of cells to IFN-γ has been shown to trigger the expression of proteins with broad antiviral effector functions, most of which act to prevent viral transcription or translation. Interestingly, in this study, we show that infection is blocked at the early step of virus entry into the host cell by retention of the minor capsid protein, L2, and the viral genome instead of trafficking into the nucleus. Thus, a novel antiviral mechanism for IFN-γ has been revealed.
Influenza A Virus Virulence Depends on Two Amino Acids in the N-Terminal Domain of Its NS1 Protein To Facilitate Inhibition of the RNA-Dependent Protein Kinase PKR
Authors: Schierhorn Kristina L. , Jolmes Fabian , Bespalowa Julia , Saenger Sandra , Peteranderl Christin , Dzieciolowski Julia , Mielke Maja , Budt Matthias , Pleschka Stephan , Herrmann Andreas , Herold Susanne , Wolff Thorsten
Published: March 2017

The RNA-dependent protein kinase (PKR) has broad antiviral activity inducing translational shutdown of viral and cellular genes and is therefore targeted by various viral proteins to facilitate pathogen propagation. The pleiotropic NS1 protein of influenza A virus acts as silencer of PKR activation and ensures high-level viral replication and virulence. However, the exact manner of this inhibition remains controversial. To elucidate the structural requirements within the NS1 protein for PKR inhibition, we generated a set of mutant viruses, identifying highly conserved arginine residues 35 and 46 within the NS1 N terminus as being most critical not only for binding to and blocking activation of PKR but also for efficient virus propagation. Biochemical and Förster resonance energy transfer (FRET)-based interaction studies showed that mutation of R35 or R46 allowed formation of NS1 dimers but eliminated any detectable binding to PKR as well as to double-stranded RNA (dsRNA). Using
in vitro
and
in vivo
approaches to phenotypic restoration, we demonstrated the essential role of the NS1 N terminus for blocking PKR. The strong attenuation conferred by NS1 mutation R35A or R46A was substantially alleviated by stable knockdown of PKR in human cells. Intriguingly, both NS1 mutant viruses did not trigger any signs of disease in PKR
+/+
mice, but replicated to high titers in lungs of PKR
−/−
mice and caused lethal infections. These data not only establish the NS1 N terminus as highly critical for neutralization of PKR's antiviral activity but also identify this blockade as an indispensable contribution of NS1 to the viral life cycle.

IMPORTANCE
Influenza A virus inhibits activation of the RNA-dependent protein kinase (PKR) by means of its nonstructural NS1 protein, but the underlying mode of inhibition is debated. Using mutational analysis, we identified arginine residues 35 and 46 within the N-terminal NS1 domain as highly critical for binding to and functional silencing of PKR. In addition, our data show that this is a main activity of amino acids 35 and 46, as the strong attenuation of corresponding mutant viruses in human cells was rescued to a large extent by lowering of PKR expression levels. Significantly, this corresponded with restoration of viral virulence for NS1 R35A and R46A mutant viruses in PKR
−/−
mice. Therefore, our data establish a model in which the NS1 N-terminal domain engages in a binding interaction to inhibit activation of PKR and ensure efficient viral propagation and virulence.
MicroRNA miR-126-5p Enhances the Inflammatory Responses of Monocytes to Lipopolysaccharide Stimulation by Suppressing Cylindromatosis in Chronic HIV-1 Infection
Authors: Huang Jun , Zhu Lingyan , Qiu Chao , Xu Xuan , Zhang Linxia , Ding Xiangqing , Liao Qibin , Xu Jianqing , Zhang Xiaoyan
Published: March 2017

Persistent immune activation during chronic human immunodeficiency virus type 1 (HIV-1) infection facilitates immune dysfunction and thereby fuels disease progression. The translocation of bacterial derivatives into blood and the hyperinflammatory responsiveness of monocytes have been considered important causative factors for persistent immune activation. Whether microRNAs (miRNAs) are involved in regulating monocyte-mediated inflammatory responses during chronic HIV-1 infection remains elusive. In this study, we show that miR-126-5p functions as a positive regulator of monocyte-mediated inflammatory responses. Significantly increased miRNA miR-126-5p and decreased cylindromatosis (CYLD) were observed in primary monocytes from chronic HIV-1 patients. Inhibition of miR-126-5p in monocytes from chronic HIV-1 patients attenuated the responsiveness of these cells to lipopolysaccharide (LPS) stimulation. Gain-of-function assays confirmed that miR-126-5p could downregulate CYLD, which in turn caused an upregulation of phosphorylation of JNK protein (pJNK) and enhanced inflammatory responses of monocytes to LPS stimulation. Overall, miR-126-5p upregulates the responsiveness of monocytes to LPS stimulation in chronic HIV-1 infection, and the suppression of miR-126-5p and the promotion of CYLD expression in primary monocytes may represent a practical immune intervention strategy to contain persistent inflammation in chronic HIV-1 infection.

IMPORTANCE
Monocyte-mediated hyperinflammatory responses during chronic HIV-1 infection are important causative factors driving AIDS progression; however, the underlying mechanism has not been fully addressed. We demonstrated that miR-126-5p, one of the most upregulated miRNAs during chronic HIV-1 infection, could enhance the inflammatory responses of monocytes to LPS by suppressing the inhibitory protein CYLD and thereby unleashing the expression of pJNK in the LPS/Toll-like receptor 4/mitogen-activated protein kinase pathway. This observation reveals a new mechanism for HIV-1 pathogenesis, which could be targeted by immune intervention.
A Respiratory Syncytial Virus Vaccine Vectored by a Stable Chimeric and Replication-Deficient Sendai Virus Protects Mice without Inducing Enhanced Disease
Authors: Wiegand Marian Alexander , Gori-Savellini Gianni , Gandolfo Claudia , Papa Guido , Kaufmann Christine , Felder Eva , Ginori Alessandro , Disanto Maria Giulia , Spina Donatella , Cusi Maria Grazia
Published: March 2017

Respiratory syncytial virus (RSV) is a major cause of severe respiratory infections in children and elderly people, and no marketed vaccine exists. In this study, we generated and analyzed a subunit vaccine against RSV based on a novel genome replication-deficient Sendai virus (SeV) vector. We inserted the RSV F protein, known to be a genetically stable antigen, into our vector in a specific way to optimize the vaccine features. By exchanging the ectodomain of the SeV F protein for its counterpart from RSV, we created a chimeric vectored vaccine that contains the RSV F protein as an essential structural component. In this way, the antigen is actively expressed on the surfaces of vaccine particles in its prefusion conformation, and as recently reported for other vectored vaccines, the occurrence of silencing mutations of the transgene in the vaccine genome can be prevented. In addition, its active gene expression contributes to further stimulation of the immune response. In order to understand the best route of immunization, we compared vaccine efficacies after intranasal (i.n.) or intramuscular (i.m.) immunization of BALB/c mice. Via both routes, substantial RSV-specific immune responses were induced, consisting of serum IgG and neutralizing antibodies, as well as cytotoxic T cells. Moreover, i.n. immunization was also able to stimulate specific mucosal IgA in the upper and lower respiratory tract. In virus challenge experiments, animals were protected against RSV infection after both i.n. and i.m. immunization without inducing vaccine-enhanced disease. Above all, the replication-deficient SeV appeared to be safe and well tolerated.

IMPORTANCE
Respiratory syncytial virus (RSV) is a major cause of respiratory diseases in young children and elderly people worldwide. There is a great demand for a licensed vaccine. Promising existing vaccine approaches based on live-attenuated vaccines or viral vectors have suffered from unforeseen drawbacks related to immunogenicity and attenuation. We provide a novel RSV vaccine concept based on a genome replication-deficient Sendai vector that has many favorable vaccine characteristics. The specific vaccine design guarantees genetic stability of the transgene; furthermore, it supports a favorable presentation of the antigen, activating the adaptive response, features that other vectored vaccine approaches have often had difficulties with. Wide immunological and pathological analyses in mice confirmed the validity and efficacy of this approach after both parenteral and mucosal administration. Above all, this concept is suitable for initiating clinical studies, and it could also be applied to other infectious diseases.
A Neonatal Murine Model of Coxsackievirus A6 Infection for Evaluation of Antiviral and Vaccine Efficacy
Authors: Zhang Zhenjie , Dong Zhaopeng , Wei Qingjuan , Carr Michael J. , Li Juan , Ding Shujun , Tong Yigang , Li Dong , Shi Weifeng
Published: March 2017

Hand, foot, and mouth disease (HFMD) is a global health concern. Family
Picornaviridae
members, particularly enterovirus A71 (EVA71) and coxsackievirus A16 (CVA16), are the primary etiological agents of HFMD; however, a third enterovirus A species, CVA6, has been recently associated with epidemic outbreaks. Study of the pathogenesis of CVA6 infection and development of antivirals and vaccines are hindered by a lack of appropriate animal models. We have developed and characterized a murine model of CVA6 infection that was employed to evaluate the antiviral activities of different drugs and the protective efficacies of CVA6-inactivated vaccines. Neonatal mice were susceptible to CVA6 infection via intramuscular inoculation, and the susceptibility of mice to CVA6 infection was age and dose dependent. Five-day-old mice infected with 10
5.5
50% tissue culture infective doses of the CVA6 WF057R strain consistently exhibited clinical signs, including reduced mobility, lower weight gain, and quadriplegia with significant pathology in the brain, hind limb skeletal muscles, and lungs of the infected mice in the moribund state. Immunohistochemical analysis and quantitative reverse transcription-PCR (qRT-PCR) analyses showed high viral loads (11 log
10
/mg) in skeletal muscle, and elevated levels of interleukin-6 (IL-6; >2,000 pg/ml) were associated with severe viral pneumonia and encephalitis. Ribavirin and gamma interferon administered prophylactically diminished CVA6-associated pathology
in vivo
, and treatment with IL-6 accelerated the death of neonatal mice. Both specific anti-CVA6 serum and maternal antibody play important roles in controlling CVA6 infection and viral replication. Collectively, these findings indicate that this neonatal murine model will be invaluable in future studies to develop CVA6-specific antivirals and vaccines.

IMPORTANCE
Although coxsackievirus A6 (CVA6) infections are commonly mild and self-limiting, a small proportion of children may have serious complications, such as encephalitis, acute flaccid paralysis, and neurorespiratory syndrome, leading to fatalities. We have established a mouse model of CVA6 infection by inoculation of neonatal mice with a CVA6 clinical isolate that produced consistent pathological outcomes. Here, using this model of CVA6 infection, we found that high levels of IL-6 were associated with severe viral pneumonia and encephalitis, as in an evaluation of antiviral efficacy
in vivo
, IL-6 had no protective effect and instead accelerated death in neonatal mice. We demonstrated that, as antiviral drugs, both gamma interferon and ribavirin played important protective roles in the early stages of infection, with increased survival in treated neonatal mice challenged with CVA6. Moreover, active and passive immunization with the inactivated vaccines and anti-CVA6 serum also protected mice against homologous challenge infections.
Attenuated Human Parainfluenza Virus Type 1 Expressing Ebola Virus Glycoprotein GP Administered Intranasally Is Immunogenic in African Green Monkeys
Authors: Lingemann Matthias , Liu Xueqiao , Surman Sonja , Liang Bo , Herbert Richard , Hackenberg Ashley D. , Buchholz Ursula J. , Collins Peter L. , Munir Shirin
Published: March 2017

The recent 2014-2016 Ebola virus (EBOV) outbreak prompted increased efforts to develop vaccines against EBOV disease. We describe the development and preclinical evaluation of an attenuated recombinant human parainfluenza virus type 1 (rHPIV1) expressing the membrane-anchored form of EBOV glycoprotein GP, as an intranasal (i.n.) EBOV vaccine. GP was codon optimized and expressed either as a full-length protein or as an engineered chimeric form in which its transmembrane and cytoplasmic tail (TMCT) domains were replaced with those of the HPIV1 F protein in an effort to enhance packaging into the vector particle and immunogenicity. GP was inserted either preceding the N gene (pre-N) or between the N and P genes (N-P) of rHPIV1 bearing a stabilized attenuating mutation in the P/C gene (C
Δ170
). The constructs grew to high titers and efficiently and stably expressed GP. Viruses were attenuated, replicating at low titers over several days, in the respiratory tract of African green monkeys (AGMs). Two doses of candidates expressing GP from the pre-N position elicited higher GP neutralizing serum antibody titers than the N-P viruses, and unmodified GP induced higher levels than its TMCT counterpart. Unmodified EBOV GP was packaged into the HPIV1 particle, and the TMCT modification did not increase packaging or immunogenicity but rather reduced the stability of GP expression during
in vivo
replication. In conclusion, we identified an attenuated and immunogenic i.n. vaccine candidate expressing GP from the pre-N position. It is expected to be well tolerated in humans and is available for clinical evaluation.

IMPORTANCE
EBOV hemorrhagic fever is one of the most lethal viral infections and lacks a licensed vaccine. Contact of fluids from infected individuals, including droplets or aerosols, with mucosal surfaces is an important route of EBOV spread during a natural outbreak, and aerosols also might be exploited for intentional virus spread. Therefore, vaccines that protect against mucosal as well as systemic inoculation are needed. We evaluated a version of human parainfluenza virus type 1 (HPIV1) bearing a stabilized attenuating mutation in the P/C gene (C
Δ170
) as an intranasal vaccine vector to express the EBOV glycoprotein GP. We evaluated expression from two different genome positions (pre-N and N-P) and investigated the use of vector packaging signals. African green monkeys immunized with two doses of the vector expressing GP from the pre-N position developed high titers of GP neutralizing serum antibodies. The attenuated vaccine candidate is expected to be safe and immunogenic and is available for clinical development.
Staurosporine Induces Filamentation in the Human Fungal Pathogen Candida albicans via Signaling through Cyr1 and Protein Kinase A
Authors: Xie Jinglin L. , O’Meara Teresa R. , Polvi Elizabeth J. , Robbins Nicole , Cowen Leah E.
Published: March 2017

The impact of fungal pathogens on human health is devastating. One of the most pervasive fungal pathogens is
Candida albicans
, which kills ~40% of people suffering from bloodstream infections. Treatment of these infections is extremely difficult, as fungi are closely related to humans, and there are limited drugs that kill the fungus without host toxicity. The capacity of
C. albicans
to transition between yeast and filamentous forms is a key virulence trait. Thus, understanding the genetic pathways that regulate morphogenesis could provide novel therapeutic targets to treat
C. albicans
infections. Here, we establish the small molecule staurosporine as an inducer of filamentous growth. We unveil distinct regulatory circuitry required for staurosporine-induced filamentation that appears to be unique to this filament-inducing cue. Thus, this work highlights the fact that small molecules, such as staurosporine, can improve our understanding of the pathways required for key virulence programs, which may lead to the development of novel therapeutics.
Environmental Viral Genomes Shed New Light on Virus-Host Interactions in the Ocean
Authors: Nishimura Yosuke , Watai Hiroyasu , Honda Takashi , Mihara Tomoko , Omae Kimiho , Roux Simon , Blanc-Mathieu Romain , Yamamoto Keigo , Hingamp Pascal , Sako Yoshihiko , Sullivan Matthew B. , Goto Susumu , Ogata Hiroyuki , Yoshida Takashi
Published: March 2017

Viruses are diverse and play significant ecological roles in marine ecosystems. However, our knowledge of genome-level diversity in viruses is biased toward those isolated from few culturable hosts. Here, we determined 1,352 nonredundant complete viral genomes from marine environments. Lifting the uncertainty that clouds short incomplete sequences, whole-genome-wide analysis suggests that these environmental genomes represent hundreds of putative novel viral genera. Predicted hosts include dominant groups of marine bacteria and archaea with no isolated viruses to date. Some of the viral genomes encode many functionally related enzymes, suggesting a strong selection pressure on these marine viruses to control cellular metabolisms by accumulating genes.
Tracking the Subtle Mutations Driving Host Sensing by the Plant Pathogen
Streptomyces scabies
Authors: Jourdan Samuel , Francis Isolde M. , Deflandre Benoit , Loria Rosemary , Rigali Sébastien
Published: March 2017

The acquisition of genetic material conferring the arsenal necessary for host virulence is a prerequisite on the path to becoming a plant pathogen. More subtle mutations are also required for the perception of cues signifying the presence of the target host and optimal conditions for colonization. The decision to activate the pathogenic lifestyle is not “taken lightly” and involves efficient systems monitoring environmental conditions. But how can a pathogen trigger the expression of virulence genes in a timely manner if the main signal inducing its pathogenic behavior originates from cellulose, the most abundant polysaccharide on earth? This situation is encountered by
Streptomyces scabies
, which is responsible for common scab disease on tuber and root crops. We propose here a series of hypotheses of how
S. scabies
could optimally distinguish whether cello-oligosaccharides originate from decomposing lignocellulose (nutrient sources, saprophyte) or, instead, emanate from living and expanding plant tissue (virulence signals, pathogen) and accordingly adapt its physiological response.
Small animal vaccination: a practical guide for vets in the UK
Authors: Day Michael J.
Published: March 2017

Vaccination is an important and fundamental part of veterinary practice and in recent years there have been significant changes in recommendations for how vaccines are given. This article reviews the reasons that these changes have occurred and the scope and purpose of vaccination guidelines. The most recent guidelines were issued by the World Small Animal Veterinary Association (WSAVA) Vaccination Guidelines Group in 2015. The article discusses how veterinarians in the UK might adapt these global guidelines for national use and provides examples of how dogs and cats can best be vaccinated according to WSAVA recommendations. Vaccination should be just one part of a holistic preventive healthcare programme for pets that is most simply delivered within the framework of an annual health check consultation.
Needlestick and inoculation injuries in veterinary and animal workers
Authors: Burke Frank , Robertson Colin , Ackerman Nicola , Reilly William
Published: March 2017

Needlestick and sharps injuries are common in veterinary practice but often go unreported. In this article, the authors, all members of the Veterinary Products Committee of the Veterinary Medicines Directorate, discuss the risks for veterinary workers and suggest strategies to prevent needlestick injuries in practice.
When the client cannot pay
Authors: Fordyce Peter
Published: March 2017

THIS series gives readers the opportunity to consider and contribute to discussion of some of the ethical dilemmas that can arise in veterinary practice. Each month, a case scenario is presented, followed by discussion of some of the issues involved.

In addition, a possible way forward is suggested; however, there is rarely a cut‐and‐dried answer in such cases, and readers may wish to suggest an alternative approach. This month's dilemma, ‘When the client cannot pay’, was submitted and is discussed by Peter Fordyce. Readers with comments to contribute are invited to send them as soon as possible, so that they can be considered for publication in the next issue.

The series is being coordinated by Steven McCulloch, a practising vet with a PhD in the ethics of veterinary policy. It aims to provide a framework that will help practitioners find solutions when facing similar dilemmas.
Diary of a parliamentary intern
Authors: Ridge Anthony
Published: March 2017

Parliamentary intern Anthony Ridge considers the range of parliamentary activities that rarely make it into the popular press.
A risk-based approach to winter road surface condition classification
Authors: Fu Liping , Thakali Lalita , Kwon Tae J. , Usman Taimur
Published: March 2017

This paper presents a risk-based approach for classifying the road surface conditions of a highway network under winter weather events. A relative risk index (RRI) is developed to capture the effect of adverse weather conditions on the collision risk of a highway in reference to the normal driving conditions. Based on this index, multiple risk factors related to adverse winter weather conditions can be considered either jointly or separately. The index can also be used to aggregate different types of road conditions observed on any given route into a single class for risk-consistent condition classification and reporting. Two example applications are shown to illustrate the advantages of the proposed approach.
Solving core issues of early physics education in Pakistan
Authors: Niazi Khadija
Published: March 2017

In the past a lot of different sources have discussed statistics and the problems present in the physics world. However, we lack discussion about possible solutions to the problems. Because of this the problems remain unsolved and we don’t come up with easy and ingenuous ideas. The basic aim of this article is to brainstorm several possible solutions to one of the problems we face in the physics world. The problem we address is the lack of women entering physics in society and an overall view on how to improve the general number of people who will enter the physics world in the future. With solutions ranging from the domestic level to the government level, I hope to suggest some points that can help solve this major dilemma.
Dynamics of a two dipole-coupled atom system driven by a quantized cavity field
Authors: Zhang Wen-Xuan , Tan Lei
Published: March 2017

Based on the master-equation approach, we investigate the dynamics of two dipole-coupled atoms by examining the atomic population and entanglement evolution. We show that the population decays rapidly and the entanglement is inhibited when the pump field is weak, while for the strongly pumped case, the evolution of population and entanglement is similar to one driven in the classical field. Furthermore, the evolution of the population can be controlled by the dipole–dipole interaction and detunings.
Modified ghost fluid method on LBM with reduced spurious pressure oscillations for moving boundaries
Authors: Chen Li , Cui Haihang , Wang Lei
Published: March 2017

A modified ghost fluid (MGF) method for reducing the spurious pressure oscillations in moving boundaries is proposed on the lattice Boltzmann method (LBM). The primary cause of these oscillations is the violation of the boundary geometric conservation law for sharp-interface immersed boundary. We introduce a simple weight strategy into GF method to strictly enforce geometric conservation. The weight strategy reduces the abrupt change of the distribution function on the boundary node when passing through the moving boundary. Some simulations are shown to test the validity of the method. The results illustrate that the modified method maintains the same validity as the GF and reduces the spurious pressure oscillations near the boundaries.
Simulation study of overtaking in pedestrian flow using floor field cellular automaton model
Authors: Fu Zhijian , Xia Liang , Yang Hongtai , Liu Xiaobo , Ma Jian , Luo Lin , Yang Lizhong , Chen Junmin
Published: March 2017

Properties of pedestrian may change along the moving path, for example, as a result of fatigue or injury, which has never been properly investigated in the past research. The paper attempts to study tactical overtaking in pedestrian flow. That is difficult to be modeled using a microscopic discrete model because of the complexity of the detailed overtaking behavior, and crossing/overlaps of pedestrian routes. Thus, a multi-velocity floor field cellular automaton model explaining the detailed psychical process of overtaking decision was proposed. Pedestrian can be either in normal state or in tactical overtaking state. Without tactical decision, pedestrians in normal state are driven by the floor field. Pedestrians make their tactical overtaking decisions by evaluating the walking environment around the overtaking route (the average velocity and density around the route, visual field of pedestrian) and obstructing conditions (the distance and velocity difference between the overtaking pedestrian and the obstructing pedestrian). The effects of tactical overtaking ratio, free velocity dispersion, and visual range on fundamental diagram, conflict density, and successful overtaking ratio were explored. Besides, the sensitivity analysis of the route factor relative intensity was performed.
Low voltage electrophoresis chip with multi-segments synchronized scanning
Authors: Gu Wenwen , Wen Zhiyu , Xu Yi
Published: March 2017

For low voltage electrophoresis chip, there is always a problem that the samples are truncated and peaks are broadened, as well as longer time for separation. In this paper, a low voltage electrophoresis separation model was established, and the separation conditions were discussed. A new driving mode was proposed for applying low voltage, which was called multi-segments synchronized scanning. By using this driving mode, the reversed electric field that existed between the multi-segments can enrich samples and shorten the sample zone. The low voltage electrophoresis experiments using multi-segments synchronized scanning were carried out by home-made silicon-PDMS-based chip. The fluorescein isothiocyanate (FITC) labeled lysine and phenylalanine mixed samples with the concentration of 10[Formula: see text] mol/L were successfully separated under the optimal conditions of 10 mmol/L borax buffer (pH = 10.0), 200 V/cm separation electric field and electrode switch time of 2.5 s. The separation was completed with a resolution of 2.0, and the peak time for lysine and phenylalanine was 4 min and 6 min, respectively.
Preparation of high-oriented molybdenum thin films using DC reactive magnetronsputtering
Authors: Shang Zhengguo , Li Dongling , Yin She , Wang Shengqiang
Published: March 2017

Since molybdenum (Mo) thin film has been used widely recently, it attracts plenty of attention, like it is a good candidate of back contact material for CuIn[Formula: see text]Ga[Formula: see text]Se[Formula: see text]S[Formula: see text] (CIGSeS) solar cells development; thanks to its more conductive and higher adhesive property. Besides, molybdenum thin film is an ideal material for aluminum nitride (AlN) thin film preparation and attributes to the tiny (−1.0%) lattice mismatch between Mo and AlN. As we know that the quality of Mo thin film is mainly dependent on process conditions, it brings a practical significance to study the influence of process parameters on Mo thin film properties. In this work, various sputtering conditions are employed to explore the feasibility of depositing a layer of molybdenum film with good quality by DC reactive magnetron sputtering. The influence of process parameters such as power, gas flow, substrate temperature and process time on the crystallinity and crystal orientation of Mo thin films is investigated. X-ray diffraction (XRD) measurements and atomic force microscope (AFM) are used to characterize the properties and surface roughness, respectively. According to comparative analysis on the results, process parameters are optimized. The full width at half maximum (FWHM) of the rocking curves of the (110) Mo is decreased to 2.7[Formula: see text], and the (110) Mo peaks reached [Formula: see text] counts. The grain size and the surface roughness have been measured as 20 Å and 3.8 nm, respectively, at 200[Formula: see text]C.
Generating the mapping class groups by torsions
Authors: Du Xiaoming
Published: March 2017

Let [Formula: see text] be a closed oriented surface of genus [Formula: see text] and let [Formula: see text] be the mapping class group. When the genus is at least 3, [Formula: see text] can be generated by torsion elements. We prove the following results: For [Formula: see text], [Formula: see text] can be generated by four torsion elements. Three generators are involutions and the fourth one is an order three element. [Formula: see text] can be generated by five torsion elements. Four generators are involutions and the fifth one is an order three element.
Intelligent Controller Based Wide-Area Control in Power System
Authors: Abedinia Oveis , Bekravi Masoud , Ghadimi Noradin
Published: March 2017

A new strategy based on Takagi–Sugeno fuzzy controller is proposed in this paper for wide-area power system. In this strategy, the proposed fuzzy controller is optimized by a new hybrid metaheuristic algorithm and hyperplane clustering algorithm to partition the fuzzy space of the given input–output data. A global signal from the centralized controller is employed in wide area control (WAC) scheme to damp out the inter-area mode as well as local mode of oscillations. To demonstrate the capability of proposed strategy, three case studies have been used in this paper. The obtained results demonstrate the validity of proposed strategy through comparison with other techniques in this problem.
SURFACE MODIFICATION OF 316L STAINLESS STEEL BY PLASMA-ASSISTED LOW TEMPERATURE CARBURIZING PROCESS
Authors: SARAVANAN M. , VENKATESHWARAN N. , DEVARAJU A. , KRISHNAKUMARI A. , SAARVESH J.
Published: March 2017

This paper aims at improving the hardness and wear resistance of Austenitic 316L Stainless Steel (SS) by Plasma-assisted Low Temperature Carburizing (PLTC) process. The process has been employed in austenitic 316L SS for achieving carbon supersaturated phase, the so-called “S Phase”. The microstructure of the treated specimens was characterized by Optical microscopy, Scanning Electron Microscopy (SEM) and X-Ray Diffraction (XRD). The results showed evidences of expanded austenite phase and formation of “S phase” at a temperature of 460[Formula: see text]C with 10% of methane (CH[Formula: see text] and 90% of hydrogen (H[Formula: see text] at a pressure of 1mbar for a time period of 20[Formula: see text]h. The hardness of the specimen was evaluated as 1030[Formula: see text]HV using Vickers microhardness setup. The wear behavior of plasma treated specimen was studied using pin on disc test at ambient condition and the results are discussed. Wear rate in PLTC 316L SS was observed to be low when compared with the wear rate of the untreated 316L SS specimen. The PLTC 316L SS specimen is subjected to ASTM A262 oxalic acid etch test to study the intergranular corrosion behavior. The “step” formation was observed in the SEM micrographs which reveal the retention of corrosion resistance in the specimen.