bluuebunny/crossref_metadata_2025 huggingface dataset:
Forty Years of Leadership Research in Sport Management: A Review, Synthesis, and Conceptual Framework
Authors: Peachey Jon Welty , Zhou Yilun , Damon Zack J. , Burton Laura J.
Published: January 2015

Scholars have recognized the importance of leadership in the sport industry; early sport leadership studies emerged in the 1970s. To date, however, there has been no comprehensive review of the scholarly leadership studies in sport management. Thus, the purpose of this review is to provide a comprehensive synthesis of the sport management leadership literature from the 1970s to the present day, to outline what has been learned, and then, drawing from this synthesis, to articulate a preliminary conceptual model capturing how leadership operates in sport management. A number of clear themes in sport management leadership research and conceptual thinking have emerged, with the proposed conceptual model advancing several leadership antecedents and processes unique to sport. Intriguing directions for sport management leadership scholarship are also illuminated. Although progress has been made, many questions and gaps remain that require focused attention from sport management leadership scholars.
Charge ordering in the electron-doped superconductor Nd
2–

x

Ce

x

CuO
4
Authors: da Silva Neto Eduardo H. , Comin Riccardo , He Feizhou , Sutarto Ronny , Jiang Yeping , Greene Richard L. , Sawatzky George A. , Damascelli Andrea
Published: January 2015

Finding order in exotic superconductors

Physicists can coax some copper-oxide compounds into becoming superconducting by chemically adding extra charge carriers: holes or electrons. Concentrating on hole-doped materials, researchers have found a host of different phases in the neighborhood of or co-existing with superconductivity. One such phase is a modulation in charge density [a charge density wave (CDW)] that appears to be ubiquitous in hole-doped families. Da Silva Neto
et al.
now show that a similar phase exists in the electron-doped material Nd

2-
x

Ce

x

CuO
4
. As they cooled the material, the authors first detected the CDW at temperatures considerably higher than in the hole-doped copper-oxides.

Science
, this issue p.
282
The roller coaster flight strategy of bar-headed geese conserves energy during Himalayan migrations
Authors: Bishop C. M. , Spivey R. J. , Hawkes L. A. , Batbayar N. , Chua B. , Frappell P. B. , Milsom W. K. , Natsagdorj T. , Newman S. H. , Scott G. R. , Takekawa J. Y. , Wikelski M. , Butler P. J.
Published: January 2015

Geese need to hug the land to fly high

Animal migrations provide numerous examples of astonishing feats. Impressive even among these is the migration of bar-headed geese across the Himalayan Mountains, which reach heights of thousands of meters. Bishop
et al.
remotely monitored birds' heart rates, movement, and body temperature during migration. The geese “hug” the landforms, taking advantage of drafting and wind patterns. This unexpected strategy conserves energy, even though it means the geese repeatedly lose, and must then regain, altitude.

Science
, this issue p.
250
Disclosing patents' secrets
Authors: Graham Stuart , Hegde Deepak
Published: January 2015

Inventors prefer to disclose know-how before patent grant
High hopes for Guinean vaccine trial
Authors: Enserink Martin
Published: January 2015

In nation with unrelenting epidemic, an unusual trial may provide key answers.
All in the (bigger) family
Authors: Pennisi Elizabeth
Published: January 2015

Revised arthropod tree marries crustacean and insect fields.
The cancer stem cell gamble
Authors: Kaiser Jocelyn
Published: January 2015

Researchers are betting that a round of clinical trials will prove a controversial cancer theory and deliver new treatments.
Vaccine-elicited CD4 T cells induce immunopathology after chronic LCMV infection
Authors: Penaloza-MacMaster Pablo , Barber Daniel L. , Wherry E. John , Provine Nicholas M. , Teigler Jeffrey E. , Parenteau Lily , Blackmore Stephen , Borducchi Erica N. , Larocca Rafael A. , Yates Kathleen B. , Shen Hao , Haining W. Nicholas , Sommerstein Rami , Pinschewer Daniel D. , Ahmed Rafi , Barouch Dan H.
Published: January 2015

For vaccines, CD4
+
T cells can spell trouble

The ideal vaccine elicits immune memory—either antibodies or memory T cells—to protect the host from subsequent infections. T cell–mediated immunity requires both helper CD4
+
T cells and cytotoxic CD8
+
T cells to kill virus-infected cells. But what happens when a vaccine only elicits CD4
+
memory T cells? Penaloza-MacMaster
et al.
probed this question by giving mice a vaccine that generated only memory CD4
+
T cells against lymphocytic choriomeningitis virus (LCMV). Instead of protecting mice against chronic LCMV, vaccinated mice developed massive inflammation and died. Virus-specific CD8
+
T cells or antibodies protected mice from the pathology. These results may have implications for vaccines against chronic viruses such as HIV.

Science
, this issue p.
278
The birth of the geological map
Authors: Sharpe Tom
Published: January 2015

Two hundred years ago, William Smith published the first geological map of a country
Semiconductor double quantum dot micromaser
Authors: Liu Y.-Y. , Stehlik J. , Eichler C. , Gullans M. J. , Taylor J. M. , Petta J. R.
Published: January 2015

Tunnel through and emit coherently

The generation of coherent light (lasers and masers) forms the basis of a large optics industry. Liu
et al.
demonstrate a type of laser that is driven by the tunneling of single electrons in semiconductor double-quantum dots. Distinct from other existing semiconductor lasers, the emission mechanism is driven by tunneling of single charges between discrete energy levels that are electrically tunable. The ability to tune the levels by single-electron charging would allow their laser (or maser) to be turned on and off rapidly.

Science
, this issue p.
285
New SQUID on the Bloch
Authors: Lamacraft Austen
Published: January 2015

A cloud of cold atoms can operate as an ultrasensitive interferometer

[Also see Report by

Duca
et al.

]
Interfering with interferons
Authors: Wilks Jessica , Golovkina Tatyana
Published: January 2015

The gut microbiota helps norovirus to persist by controlling λ interferons

[Also see Reports by

Baldridge
et al.

and

Nice
et al.

]
Evaluation of a Multiplex Bead Immunoassay for Determination of Immune Status to Varicella-Zoster Virus in Medical Center Students and Employees
Authors: Loeffelholz Michael J. , Prince Harry E.
Published: January 2015

This study evaluated an enzyme immunoassay, a multiplex bead immunoassay (MBIA), and the anticomplement immunofluorescence (ACIF) test for detecting varicella-zoster virus IgG antibodies in sera from medical center students and employees. The agreement between methods was ≥95%. The MBIA was less sensitive than was the ACIF test, with a negative predictive value of 66.7%.
Genetic, Biochemical, and Serological Characterization of a New Pneumococcal Serotype, 6H, and Generation of a Pneumococcal Strain Producing Three Different Capsular Repeat Units
Authors: Park In Ho , Geno K. Aaron , Yu Jigui , Oliver Melissa B. , Kim Kyung-Hyo , Nahm Moon H.
Published: January 2015

Streptococcus pneumoniaeclinical isolates were recently described that produced capsular polysaccharide with properties of both serotypes 6A and 6B. Their hybrid serological property correlated with mutations affecting the glycosyltransferase WciP, which links rhamnose to ribitol by an α(1-3) linkage for serotypes 6A and 6C and an α(1-4) linkage for serotypes 6B and 6D. The isolates had mutations in the triad residues of WciP that have been correlated with enzyme specificity. The canonical triad residues of WciP are Ala192-Ser195-Arg254 for serotypes 6A and 6C and Ser192-Asn195-Gly254 for serotypes 6B and 6D. To prove that the mutations in the triad residues are responsible for the hybrid serotype, we introduced the previously described Ala192-Cys195-Arg254 triad into a 6A strain and found that the change made WciP bispecific, resulting in 6A and 6B repeat unit expression, although 6B repeat unit production was favored over production of 6A repeat units. Likewise, this triad permitted a 6C strain to express 6C and 6D repeat units. With reported bispecificity in WciN, which adds either glucose or galactose as the second sugar in the serogroup 6 repeat unit, the possibility exists for a strain to simultaneously produce all four serogroup 6 repeat units; however, when genes encoding both bispecific enzymes were introduced into a 6A strain, only 6A, 6B, and 6D repeat units were detected serologically. Nonetheless, this may be the first example of a bacterial polysaccharide with three different repeat units. This strategy of expressing multiple repeat units in a single polymer is a novel approach to broadening vaccine coverage by eliminating the need for multiple polysaccharide sources to cover multiple serogroup members.
Next-Generation Sequencing Reveals a Controlled Immune Response to Zaire Ebola Virus Challenge in Cynomolgus Macaques Immunized with Vesicular Stomatitis Virus Expressing Zaire Ebola Virus Glycoprotein (VSVΔG/EBOVgp)
Authors: Barrenas Fredrik , Green Richard R. , Thomas Matthew J. , Law G. Lynn , Proll Sean C. , Engelmann Flora , Messaoudi Ilhem , Marzi Andrea , Feldmann Heinz , Katze Michael G.
Published: January 2015

Vesicular stomatitis virus expressing Zaire Ebola virus (EBOV) glycoprotein (VSVΔG/EBOVgp) could be used as a vaccine to meet the 2014 Ebola virus outbreak. To characterize the host response to this vaccine, we used mRNA sequencing to analyze peripheral blood mononuclear cells (PBMCs) from cynomolgus macaques after VSVΔG/EBOVgp immunization and subsequent EBOV challenge. We found a controlled transcriptional response that transitioned to immune regulation as the EBOV was cleared. This observation supports the safety of the vaccine.
Antibody-Mediated Complement C3b/iC3b Binding to Group B Streptococcus in Paired Mother and Baby Serum Samples in a Refugee Population on the Thailand-Myanmar Border
Authors: Herbert Jenny , Thomas Stephen , Brookes Charlotte , Turner Claudia , Turner Paul , Nosten Francois , Le Doare Kirsty , Hudson Michael , Heath Paul T. , Gorringe Andrew , Taylor Stephen
Published: January 2015

Streptococcus agalactiae(group B streptococcus [GBS]) is the leading cause of neonatal sepsis and meningitis. In this study, we determined antibody-mediated deposition of complement C3b/iC3b onto the bacterial cell surface of GBS serotypes Ia, Ib, II, III, and V. This was determined for 520 mother and umbilical cord serum sample pairs obtained at the time of birth from a population on the Thailand-Myanmar border. Antibody-mediated deposition of complement C3b/iC3b was detected to at least one serotype in 91% of mothers, despite a known carriage rate in this population of only 12%. Antibody-mediated C3b/iC3b deposition corresponded to known carriage rates, with the highest levels of complement deposition observed onto the most prevalent serotype (serotype II) followed by serotypes Ia, III, V, and Ib. Finally, neonates born to mothers carrying serotype II GBS at the time of birth showed higher antibody-mediated C3b/iC3b deposition against serotype II GBS than neonates born to mothers with no serotype II carriage. Assessment of antibody-mediated C3b/iC3b deposition against GBS may provide insights into the seroepidemiology of anti-GBS antibodies in mothers and infants in different populations.
Development of a Real-Time PCR Assay for Identification of Coccidioides immitis by Use of the BD Max System
Authors: Mitchell Marilyn , Dizon Dominic , Libke Robert , Peterson Michael , Slater David , Dhillon Akashdeep
Published: January 2015

Rapid real-time PCR (RT-PCR) can be performed in a community hospital setting to identify
Coccidioides
species using the new Becton Dickinson molecular instrument BD Max. Following sample preparation, DNA extraction and PCR were performed on the BD Max using the BD Max extraction kit ExK-DNA-1 test strip and a master mix prepared by BioGX (Birmingham, AL). Sample preparation took 2 h, and testing on the BD Max took an additional 2 h. Method sensitivity and specificity were evaluated along with the limits of detection to confirm that this convenient method would provide medically useful information. Using serial dilutions, the lower limit of detection was determined to be 1 CFU/μl. Testing with this method was validated using samples from various body sites, including bronchial alveolar lavage (BAL) fluid; sputum and lung tissue samples; and pleural and spinal fluids. Safety protocols were established, and specimen preparation processes were developed for the various types of specimens. The range for the cycle threshold (

C
T

) indicating adequate fluorescent signal to signify a positive result was established along with the acceptable range for the internal standard. Positive controls run with each batch were prepared by spiking a pooled BAL fluid specimen with a known dilution of
Coccidioides immitis
organism. Our experience with testing >330 patient samples shows that clinically relevant information can be available within 4 h using an RT-PCR method on the BD Max to identify
Coccidioides
spp., with sensitivity equivalent to culture.
Multicountry Prospective Clinical Evaluation of Two Enzyme-Linked Immunosorbent Assays and Two Rapid Diagnostic Tests for Diagnosing Dengue Fever
Authors: Pal Subhamoy , Dauner Allison L. , Valks Andrea , Forshey Brett M. , Long Kanya C. , Thaisomboonsuk Butsaya , Sierra Gloria , Picos Victor , Talmage Sara , Morrison Amy C. , Halsey Eric S. , Comach Guillermo , Yasuda Chadwick , Loeffelholz Michael , Jarman Richard G. , Fernandez Stefan , An Ung Sam , Kochel Tadeusz J. , Jasper Louis E. , Wu Shuenn-Jue L.
Published: January 2015

We evaluated four dengue diagnostic devices from Alere, including the SD Bioline Dengue Duo (nonstructural [NS] 1 Ag and IgG/IgM), the Panbio Dengue Duo Cassette (IgM/IgG) rapid diagnostic tests (RDTs), and the Panbio dengue IgM and IgG capture enzyme-linked immunosorbent assays (ELISAs) in a prospective, controlled, multicenter study in Peru, Venezuela, Cambodia, and the United States, using samples from 1,021 febrile individuals. Archived, well-characterized samples from an additional 135 febrile individuals from Thailand were also used. Reference testing was performed on all samples using an algorithm involving virus isolation, in-house IgM and IgG capture ELISAs, and plaque reduction neutralization tests (PRNT) to determine the infection status of the individual. The primary endpoints were the clinical sensitivities and specificities of these devices. The SD Bioline Dengue Duo had an overall sensitivity of 87.3% (95% confidence interval [CI], 84.1 to 90.2%) and specificity of 86.8% (95% CI, 83.9 to 89.3%) during the first 14 days post-symptom onset (p.s.o.). The Panbio Dengue Duo Cassette demonstrated a sensitivity of 92.1% (87.8 to 95.2%) and specificity of 62.2% (54.5 to 69.5%) during days 4 to 14 p.s.o. The Panbio IgM capture ELISA had a sensitivity of 87.6% (82.7 to 91.4%) and specificity of 88.1% (82.2 to 92.6%) during days 4 to 14 p.s.o. Finally, the Panbio IgG capture ELISA had a sensitivity of 69.6% (62.1 to 76.4%) and a specificity of 88.4% (82.6 to 92.8%) during days 4 to 14 p.s.o. for identification of secondary dengue infections. This multicountry prospective study resulted in reliable real-world performance data that will facilitate data-driven laboratory test choices for managing patient care during dengue outbreaks.
Evaluation of Antigen-Specific Immunoglobulin G Responses in Pulmonary Tuberculosis Patients and Contacts
Authors: Hur Yun-Gyoung , Kim Ahreum , Kang Young Ae , Kim An Sik , Kim Dae Yeon , Kim Yeun , Kim Youngmi , Lee Hyeyoung , Cho Sang-Nae
Published: January 2015

This study aimed to evaluate the serodiagnostic potential of immunoglobulin G (IgG) responses to
Mycobacterium tuberculosis
antigens in pulmonary tuberculosis (TB) patients, recent TB contacts with latent TB infection (LTBI), and healthy subjects. Infections were assessed using tuberculin skin tests, QuantiFERON-TB Gold In-Tube tests, drug susceptibility testing, and molecular genotyping of clinical isolates. Serum IgG responses to selective
M. tuberculosis
antigens, including the 38-kDa and 16-kDa antigens, lipoarabinomannan (LAM), and recombinant early secreted antigen target 6 kDa (ESAT-6) and culture filtrate protein 10 kDa (CFP-10), were determined. We found that the serum IgG responses to all antigens might differentiate between active TB and LTBI, with LAM having the highest diagnostic value (area under the curve [AUC] of 0.7756,
P
< 0.001). Recurrent TB cases showed significantly higher IgG responses to 38 kDa, CFP-10 (
P
< 0.01), and LAM (
P
< 0.05) than new cases, and male patients had higher levels of antigen-specific IgG than females (
P
< 0.05). Conversely, drug resistance and patient body mass index did not affect IgG responses (
P
> 0.05). LAM-specific IgG responses differentiated between acid-fast bacillus (AFB) smear-positive and -negative patients (
P
< 0.01), whereas antigen-specific IgG responses did not vary with the
M. tuberculosis
genotype (
P
> 0.05). Significantly higher IgG responses to 38 kDa and 16 kDa were observed in AFB smear-negative patients than in controls. These results suggest that assessment of serum IgG responses to selective purified
M. tuberculosis
antigens may help improve the diagnosis of active TB, particularly for sputum smear-negative patients or recurrent cases, and these may also help to differentiate between active TB and LTBI.
Evaluation of Enzyme Immunoassays and Real-Time PCR for Detecting Shiga Toxin-Producing Escherichia coli in Southern Alberta, Canada
Authors: Chui Linda , Patterson-Fortin Laura , Kuo Julie , Li Vincent , Boras Valerie
Published: January 2015

Two immunoassays (Shiga Toxin Chek and Shiga Toxin Quik Chek) and real-time PCR were used to detect Shiga toxin-producing
Escherichia coli
. For enriched culture, the sensitivity and specificity of the three methods ranged from 80.0% to 98.2% and 98.0% to 100.0%, respectively. STEC isolates were identified in 2.6% of the 784 samples.
Development and Validation of a Quantitative PCR Assay Using Multiplexed Hydrolysis Probes for Detection and Quantification of Theileria orientalis Isolates and Differentiation of Clinically Relevant Subtypes
Authors: Bogema D. R. , Deutscher A. T. , Fell S. , Collins D. , Eamens G. J. , Jenkins C.
Published: January 2015

Theileria orientalis
is an emerging pathogen of cattle in Asia, Australia, and New Zealand. This organism is a vector-borne hemoprotozoan that causes clinical disease characterized by anemia, abortion, and death, as well as persistent subclinical infections. Molecular methods of diagnosis are preferred due to their sensitivity and utility in differentiating between pathogenic and apathogenic genotypes. Conventional PCR (cPCR) assays for
T. orientalis
detection and typing are laborious and do not provide an estimate of parasite load. Current real-time PCR assays cannot differentiate between clinically relevant and benign genotypes or are only semiquantitative without a defined clinical threshold. Here, we developed and validated a hydrolysis probe quantitative PCR (qPCR) assay which universally detects and quantifies
T. orientalis
and identifies the clinically associated Ikeda and Chitose genotypes (UIC assay). Comparison of the UIC assay results with previously validated universal and genotype-specific cPCR results demonstrated that qPCR detects and differentiates
T. orientalis
with high sensitivity and specificiy. Comparison of quantitative results based on percent parasitemia, determined via blood film analysis and packed cell volume (PCV) revealed significant positive and negative correlations, respectively. One-way analysis of variance (ANOVA) indicated that blood samples from animals with clinical signs of disease contained statistically higher concentrations of
T. orientalis
DNA than animals with subclinical infections. We propose clinical thresholds to assist in classifying high-, moderate-, and low-level infections and describe how parasite load and the presence of the Ikeda and Chitose genotypes relate to disease.
High Preexisting Serological Antibody Levels Correlate with Diversification of the Influenza Vaccine Response
Authors: Andrews Sarah F. , Kaur Kaval , Pauli Noel T. , Huang Min , Huang Yunping , Wilson Patrick C.
Published: January 2015

Reactivation of memory B cells allows for a rapid and robust immune response upon challenge with the same antigen. Variant influenza virus strains generated through antigenic shift or drift are encountered multiple times over the lifetime of an individual. One might predict, then, that upon vaccination with the trivalent influenza vaccine across multiple years, the antibody response would become more and more dominant toward strains consistently present in the vaccine at the expense of more divergent strains. However, when we analyzed the vaccine-induced plasmablast, memory, and serological responses to the trivalent influenza vaccine between 2006 and 2013, we found that the B cell response was most robust against more divergent strains. Overall, the antibody response was highest when one or more strains contained in the vaccine varied from year to year. This suggests that in the broader immunological context of viral antigen exposure, the B cell response to variant influenza virus strains is not dictated by the composition of the memory B cell precursor pool. The outcome is instead a diversified B cell response.

IMPORTANCEVaccine strategies are being designed to boost broadly reactive B cells present in the memory repertoire to provide universal protection to the influenza virus. It is important to understand how past exposure to influenza virus strains affects the response to subsequent immunizations. The viral epitopes targeted by B cells responding to the vaccine may be a direct reflection of the B cell memory specificities abundant in the preexisting immune repertoire, or other factors may influence the vaccine response. Here, we demonstrate that high preexisting serological antibody levels to a given influenza virus strain correlate with low production of antibody-secreting cells and memory B cells recognizing that strain upon revaccination. In contrast, introduction of antigenically novel strains generates a robust B cell response. Thus, both the preexisting memory B cell repertoire and serological antibody levels must be taken into consideration in predicting the quality of the B cell response to new prime-boost vaccine strategies.
Structural Basis for Host Membrane Remodeling Induced by Protein 2B of Hepatitis A Virus
Authors: Vives-Adrián Laia , Garriga Damià , Buxaderas Mònica , Fraga Joana , Pereira Pedro José Barbosa , Macedo-Ribeiro Sandra , Verdaguer Núria
Published: January 2015

The complexity of viral RNA synthesis and the numerous participating factors require a mechanism to topologically coordinate and concentrate these multiple viral and cellular components, ensuring a concerted function. Similarly to all other positive-strand RNA viruses, picornaviruses induce rearrangements of host intracellular membranes to create structures that act as functional scaffolds for genome replication. The membrane-targeting proteins 2B and 2C, their precursor 2BC, and protein 3A appear to be primarily involved in membrane remodeling. Little is known about the structure of these proteins and the mechanisms by which they induce massive membrane remodeling. Here we report the crystal structure of the soluble region of hepatitis A virus (HAV) protein 2B, consisting of two domains: a C-terminal helical bundle preceded by an N-terminally curved five-stranded antiparallel β-sheet that displays striking structural similarity to the β-barrel domain of enteroviral 2A proteins. Moreover, the helicoidal arrangement of the protein molecules in the crystal provides a model for 2B-induced host membrane remodeling during HAV infection.

IMPORTANCENo structural information is currently available for the 2B protein of any picornavirus despite it being involved in a critical process in viral factory formation: the rearrangement of host intracellular membranes. Here we present the structure of the soluble domain of the 2B protein of hepatitis A virus (HAV). Its arrangement, both in crystals and in solution under physiological conditions, can help to understand its function and sheds some light on the membrane rearrangement process, a putative target of future antiviral drugs. Moreover, this first structure of a picornaviral 2B protein also unveils a closer evolutionary relationship between the hepatovirus and enterovirus genera within thePicornaviridaefamily.
Hepatitis C Virus-Mediated Enhancement of MicroRNA miR-373 Impairs the JAK/STAT Signaling Pathway
Authors: Mukherjee Anupam , Di Bisceglie Adrian M. , Ray Ratna B.
Published: January 2015

Hepatitis C virus (HCV) is a serious global health problem and establishes chronic infection in a significant number of infected humans worldwide. Interferon (IFN) and IFN-stimulated genes (ISGs) are amplified during HCV infection but fail to eliminate virus from the liver in a large number of infected patients, and the mechanism is not fully understood. MicroRNAs (miRNAs) have been implicated in the control of many biological processes, including IFN signaling. To gain more insights into the role of cellular miRNAs in possible countermeasures of HCV for suppression of the host antiviral response, a miRNA array was performed by using primary human hepatocytes infected with
in vitro
cell culture-grown HCV. A group of miRNAs were modulated in HCV-infected primary human hepatocytes. We focused on miR-373, as this miRNA was significantly upregulated in HCV-infected primary human hepatocytes. Here, we analyzed the function of miR-373 in the context of HCV infection. HCV infection upregulates miR-373 expression in hepatocytes and HCV-infected liver biopsy specimens. Furthermore, we discovered that miR-373 directly targets Janus kinase 1 (JAK1) and IFN-regulating factor 9 (IRF9), important factors in the IFN signaling pathway. The upregulation of miR-373 by HCV also inhibited STAT1 phosphorylation, which is involved in ISG factor 3 (ISGF3) complex formation and ISG expression. The knockdown of miR-373 in hepatocytes enhanced JAK1 and IRF9 expression and reduced HCV RNA replication. Taken together, our results demonstrated that miR-373 is upregulated during HCV infection and negatively regulated the type I IFN signaling pathway by suppressing JAK1 and IRF9. Our results offer a potential therapeutic approach for antiviral intervention.

IMPORTANCE
Chronic HCV infection is one of the major causes of end-stage liver disease worldwide. Although the recent introduction of direct-acting antiviral (DAA) therapy is extremely encouraging, some infected individuals do not respond to this therapy. Furthermore, these drugs target HCV nonstructural proteins, and with selective pressure, the virus may develop a resistant strain. Therefore, understanding the impairment of IFN signals will help in designing additional therapeutic modalities. In this study, we provide evidence of HCV-mediated upregulation of miR-373 and show that miR-373 impairs IFN signaling by targeting JAK1/IRF9 molecules. The knockdown of miR-373 inhibited HCV replication by upregulating interferon-stimulating gene expression. Together, these results provided new mechanistic insights into the role of miR-373 in HCV infection and suggest a new potential target against HCV infection.
Absence of the Uracil DNA Glycosylase of Murine Gammaherpesvirus 68 Impairs Replication and Delays the Establishment of Latency
In Vivo
Authors: Minkah Nana , Macaluso Marc , Oldenburg Darby G. , Paden Clinton R. , White Douglas W. , McBride Kevin M. , Krug Laurie T.
Published: January 2015

Uracil DNA glycosylases (UNG) are highly conserved proteins that preserve DNA fidelity by catalyzing the removal of mutagenic uracils. All herpesviruses encode a viral UNG (vUNG), and yet the role of the vUNG in a pathogenic course of gammaherpesvirus infection is not known. First, we demonstrated that the vUNG of murine gammaherpesvirus 68 (MHV68) retains the enzymatic function of host UNG in an
in vitro
class switch recombination assay. Next, we generated a recombinant MHV68 with a stop codon in ORF46/UNG (ΔUNG) that led to loss of UNG activity in infected cells and a replication defect in primary fibroblasts. Acute replication of MHV68ΔUNG in the lungs of infected mice was reduced 100-fold and was accompanied by a substantial delay in the establishment of splenic latency. Latency was largely, yet not fully, restored by an increase in virus inoculum or by altering the route of infection. MHV68 reactivation from latent splenocytes was not altered in the absence of the vUNG. A survey of host UNG activity in cells and tissues targeted by MHV68 indicated that the lung tissue has a lower level of enzymatic UNG activity than the spleen. Taken together, these results indicate that the vUNG plays a critical role in the replication of MHV68 in tissues with limited host UNG activity and this vUNG-dependent expansion, in turn, influences the kinetics of latency establishment in distal reservoirs.

IMPORTANCE
Herpesviruses establish chronic lifelong infections using a strategy of replicative expansion, dissemination to latent reservoirs, and subsequent reactivation for transmission and spread. We examined the role of the viral uracil DNA glycosylase, a protein conserved among all herpesviruses, in replication and latency of murine gammaherpesvirus 68. We report that the viral UNG of this murine pathogen retains catalytic activity and influences replication in culture. The viral UNG was impaired for productive replication in the lung. This defect in expansion at the initial site of acute replication was associated with a substantial delay of latency establishment in the spleen. The levels of host UNG were substantially lower in the lung compared to the spleen, suggesting that herpesviruses encode a viral UNG to compensate for reduced host enzyme levels in some cell types and tissues. These data suggest that intervention at the site of initial replicative expansion can delay the establishment of latency, a hallmark of chronic herpesvirus infection.
Emergence and Evolution of H10 Subtype Influenza Viruses in Poultry in China
Authors: Ma Chi , Lam Tommy Tsan-Yuk , Chai Yujuan , Wang Jia , Fan Xiaohui , Hong Wenshan , Zhang Yu , Li Lifeng , Liu Yongmei , Smith David K. , Webby Richard J. , Peiris Joseph S. M. , Zhu Huachen , Guan Yi
Published: January 2015

The cases of human infections with H10N8 viruses identified in late 2013 and early 2014 in Jiangxi, China, have raised concerns over the origin, prevalence, and development of these viruses in this region. Our long-term influenza surveillance of poultry and migratory birds in southern China in the past 12 years showed that H10 influenza viruses have been introduced from migratory to domestic ducks over several winter seasons at sentinel duck farms at Poyang Lake, where domestic ducks share their water body with overwintering migratory birds. H10 viruses were never detected in terrestrial poultry in our survey areas until August 2013, when they were identified at live-poultry markets in Jiangxi. Since then, we have isolated 124 H10N8 or H10N6 viruses from chickens at local markets, revealing an ongoing outbreak. Phylogenetic analysis of H10 and related viruses showed that the chicken H10N8 viruses were generated through multiple reassortments between H10 and N8 viruses from domestic ducks and the enzootic chicken H9N2 viruses. These chicken reassortant viruses were highly similar to the human isolate, indicating that market chickens were the source of human infection. Recently, the H10 viruses further reassorted, apparently with H5N6 viruses, and generated an H10N6 variant. The emergence and prevalence of H10 viruses in chickens and the occurrence of human infections provide direct evidence of the threat from the current influenza ecosystem in China.

IMPORTANCE
After the outbreak of avian-origin H7N9 influenza viruses in China, fatal human infections with a novel H10N8 virus were reported. Utilizing data from 12 years of influenza surveillance in southern China, we showed that H10 viruses were regularly introduced by migratory ducks to domestic ducks on Poyang Lake, a major aggregative site of migratory birds in Asia. The H10 viruses were maintained and amplified in domestic ducks and then transmitted to chickens and reassorted with enzootic H9N2 viruses, leading to an outbreak and human infections at live-poultry markets. The emergence of the H10N8 virus, following a pathway similar to that of the recent H7N9 virus, highlights the role of domestic ducks and the current influenza ecosystem in China that facilitates influenza viruses moving from their reservoir hosts through the live-poultry system to cause severe consequences for public health.
Mining the Human Complexome Database Identifies RBM14 as an XPO1-Associated Protein Involved in HIV-1 Rev Function
Authors: Budhiraja Sona , Liu Hongbing , Couturier Jacob , Malovannaya Anna , Qin Jun , Lewis Dorothy E. , Rice Andrew P.
Published: January 2015

By recruiting the host protein XPO1 (CRM1), the HIV-1 Rev protein mediates the nuclear export of incompletely spliced viral transcripts. We mined data from the recently described human nuclear complexome to identify a host protein, RBM14, which associates with XPO1 and Rev and is involved in Rev function. Using a Rev-dependent p24 reporter plasmid, we found that RBM14 depletion decreased Rev activity and Rev-mediated enhancement of the cytoplasmic levels of unspliced viral transcripts. RBM14 depletion also reduced p24 expression during viral infection, indicating that RBM14 is limiting for Rev function. RBM14 has previously been shown to localize to nuclear paraspeckles, a structure implicated in retaining unspliced HIV-1 transcripts for either Rev-mediated nuclear export or degradation. We found that depletion of NEAT1 RNA, a long noncoding RNA required for paraspeckle integrity, abolished the ability of overexpressed RBM14 to enhance Rev function, indicating the dependence of RBM14 function on paraspeckle integrity. Our study extends the known host cell interactome of Rev and XPO1 and further substantiates a critical role for paraspeckles in the mechanism of action of Rev. Our study also validates the nuclear complexome as a database from which viral cofactors can be mined.

IMPORTANCE
This study mined a database of nuclear protein complexes to identify a cellular protein named RBM14 that is associated with XPO1 (CRM1), a nuclear protein that binds to the HIV-1 Rev protein and mediates nuclear export of incompletely spliced viral RNAs. Functional assays demonstrated that RBM14, a protein found in paraspeckle structures in the nucleus, is involved in HIV-1 Rev function. This study validates the nuclear complexome database as a reference that can be mined to identify viral cofactors.
Conserved Molecular Signatures in gp120 Are Associated with the Genetic Bottleneck during Simian Immunodeficiency Virus (SIV), SIV-Human Immunodeficiency Virus (SHIV), and HIV Type 1 (HIV-1) Transmission
Authors: Gonzalez Mileidy W. , DeVico Anthony L. , Lewis George K. , Spouge John L.
Published: January 2015

Human immunodeficiency virus (HIV) transmission typically results from infection by a single transmitted/founder (T/F) variant. Are T/F variants chosen uniformly at random from the donor pool, or are they selected based on advantageous traits facilitating transmission? Finding evidence for selection during transmission is of particular interest, because it would indicate that phenotypic and/or genetic properties of the viruses might be harnessed as potential vaccine targets or immunotherapies. Here, we systematically evaluated the differences between the Env proteins of simian immunodeficiency virus/simian HIV (SIV/SHIV) stock and T/F variants in search of “signature” sites of transmission. We also surveyed residue preferences in HIV at the SIV/SHIV signature sites. Four sites of gp120 showed significant selection, and an additional two sites showed a similar trend. Therefore, the six sites clearly differentiate T/F viruses from the majority of circulating variants in the stocks. The selection of SIV/SHIV could be inferred reasonably across both vaccinated and unvaccinated subjects, with infections resulting from vaginal, rectal, and intravenous routes of transmission and regardless of viral dosage. The evidence for selection in SIV and SHIV T/F variants is strong and plentiful, and in HIV the evidence is suggestive though commensurate with the availability of suitable data for analysis. Two of the signature residues are completely conserved across the SIV, SHIV, and HIV variants we examined. Five of the signature residues map to the C1 region of gp120 and one to the signal peptide. Our data raise the possibility that C1, while governing the association between gp120 and gp41, modulates transmission efficiency, replicative fitness, and/or host cell tropism at the level of virus-cell attachment and entry.

IMPORTANCE
The present study finds significant evidence of selection on gp120 molecules of SIV/SHIV T/F viruses. The data provide ancillary evidence suggesting the same sites are under selection in HIV. Our findings suggest that the signature residues are involved in increasing the transmissibility of infecting viruses; therefore, they are potential targets for developing a vaccine or other protective measures. A recent study identified the same T/F signature motif but interpreted it as an effect of neutralization resistance. Here, we show that the T/F motif has broader functional significance beyond neutralization sensitivity, because it is present in nonimmune subjects. Also, a vaccine regimen popular in animal trials might have increased the transmission of variants with otherwise low transmission fitness. Our observations might explain why many animal vaccine trials have not faithfully predicted outcomes in human vaccine trials and suggest that current practices in vaccine design need to be reexamined accordingly.
Postchallenge Administration of Brincidofovir Protects Healthy and Immune-Deficient Mice Reconstituted with Limited Numbers of T Cells from Lethal Challenge with IHD-J-Luc Vaccinia Virus
Authors: Zaitseva Marina , McCullough Kevin Tyler , Cruz Stephanie , Thomas Antonia , Diaz Claudia G. , Keilholz Laurie , Grossi Irma M. , Trost Lawrence C. , Golding Hana
Published: January 2015

Protection from lethality by postchallenge administration of brincidofovir (BCV, CMX001) was studied in normal and immune-deficient (nude, nu/nu) BALB/c mice infected with vaccinia virus (VACV). Whole-body bioluminescence imaging was used to record total fluxes in the nasal cavity, lungs, spleen, and liver and to enumerate pox lesions on tails of mice infected via the intranasal route with 10
5
PFU of recombinant IHD-J-Luc VACV expressing luciferase. Areas under the flux curve (AUCs) were calculated for individual mice to assess viral loads. A three-dose regimen of 20 mg/kg BCV administered every 48 h starting either on day 1 or day 2 postchallenge protected 100% of mice. Initiating BCV treatment earlier was more efficient in reducing viral loads and in providing protection from pox lesion development. All BCV-treated mice that survived challenge were also protected from rechallenge with IHD-J-Luc or WRvFire VACV without additional treatment. In immune-deficient mice, BCV protected animals from lethality and reduced viral loads while animals were on the drug. Viral recrudescence occurred within 4 to 9 days, and mice succumbed ∼10 to 20 days after treatment termination. Nude mice reconstituted with 10
5
T cells prior to challenge with 10
4
PFU of IHD-J-Luc and treated with BCV postchallenge survived the infection, cleared the virus from all organs, and survived rechallenge with 10
5
PFU of IHD-J-Luc VACV without additional BCV treatment. Together, these data suggest that BCV protects immunocompetent and partially T cell-reconstituted immune-deficient mice from lethality, reduces viral dissemination in organs, prevents pox lesion development, and permits generation of VACV-specific memory.

IMPORTANCE
Mass vaccination is the primary element of the public health response to a smallpox outbreak. In addition to vaccination, however, antiviral drugs are required for individuals with uncertain exposure status to smallpox or for whom vaccination is contraindicated. Whole-body bioluminescence imaging was used to study the effect of brincidofovir (BCV) in normal and immune-deficient (nu/nu) mice infected with vaccinia virus, a model of smallpox. Postchallenge administration of 20 mg/kg BCV rescued normal and immune-deficient mice partially reconstituted with T cells from lethality and significantly reduced viral loads in organs. All BCV-treated mice that survived infection were protected from rechallenge without additional treatment. In immune-deficient mice, BCV extended survival. The data show that BCV controls viral replication at the site of challenge and reduces viral dissemination to internal organs, thus providing a shield for the developing adaptive immunity that clears the host of virus and builds virus-specific immunological memory.
Mapping the Interactions between the NS4B and NS3 Proteins of Dengue Virus
Authors: Zou Jing , Lee Le Tian , Wang Qing Yin , Xie Xuping , Lu Siyan , Yau Yin Hoe , Yuan Zhiming , Geifman Shochat Susana , Kang Congbao , Lescar Julien , Shi Pei-Yong
Published: January 2015

Flavivirus RNA synthesis is mediated by a multiprotein complex associated with the endoplasmic reticulum membrane, named the replication complex (RC). Within the flavivirus RC, NS4B, an integral membrane protein with a role in virulence and regulation of the innate immune response, binds to the NS3 protease-helicase. NS4B modulates the RNA helicase activity of NS3, but the molecular details of their interaction remain elusive. Here, we used dengue virus (DENV) to map the determinants for the NS3-NS4B interaction. Coimmunoprecipitation and an
in situ
proximity ligation assay confirmed that NS3 colocalizes with NS4B in both DENV-infected cells and cells coexpressing both proteins. Surface plasmon resonance demonstrated that subdomains 2 and 3 of the NS3 helicase region and the cytoplasmic loop of NS4B are required for binding. Using nuclear magnetic resonance (NMR), we found that the isolated cytoplasmic loop of NS4B is flexible, with a tendency to form a three-turn α-helix and two short β-strands. Upon binding to the NS3 helicase, 12 amino acids within the cytoplasmic loop of NS4B exhibited line broadening, suggesting a participation in the interaction. Sequence alignment showed that 4 of these 12 residues are strictly conserved across different flaviviruses. Mutagenesis analysis showed that three (Q134, G140, and N144) of the four evolutionarily conserved NS4B residues are essential for DENV replication. The mapping of the NS3/NS4B-interacting regions described here can assist the design of inhibitors that disrupt their interface for antiviral therapy.

IMPORTANCE
NS3 and NS4B are essential components of the flavivirus RC. Using DENV as a model, we mapped the interaction between the viral NS3 and NS4B proteins. The subdomains 2 and 3 of NS3 helicase as well as the cytoplasmic loop of NS4B are critical for the interaction. Functional analysis delineated residues within the NS4B cytoplasmic loop that are crucial for DENV replication. Our findings reveal molecular details of how flavivirus NS3 protein cooperates with NS4B within the RC. In addition, this study has established the rationale and assays to search for inhibitors disrupting the NS3-NS4B interaction for antiviral drug discovery.
Early Function of the Abutilon Mosaic Virus AC2 Gene as a Replication Brake
Authors: Krenz Björn , Deuschle Kathrin , Deigner Tobias , Unseld Sigrid , Kepp Gabi , Wege Christina , Kleinow Tatjana , Jeske Holger
Published: January 2015

The C2/AC2 genes of monopartite/bipartite geminiviruses of the genera
Begomovirus
and
Curtovirus
encode important pathogenicity factors with multiple functions described so far. A novel function of
Abutilon
mosaic virus (AbMV) AC2 as a replication brake is described, utilizing transgenic plants with dimeric inserts of DNA B or with a reporter construct to express green fluorescent protein (GFP). Their replicational release upon AbMV superinfection or the individual and combined expression of epitope-tagged AbMV AC1, AC2, and AC3 was studied. In addition, the effects were compared in the presence and in the absence of an unrelated tombusvirus suppressor of silencing (P19). The results show that AC2 suppresses replication reproducibly in all assays and that AC3 counteracts this effect. Examination of the topoisomer distribution of supercoiled DNA, which indicates changes in the viral minichromosome structure, did not support any influence of AC2 on transcriptional gene silencing and DNA methylation. The geminiviral AC2 protein has been detected here for the first time in plants. The experiments revealed an extremely low level of AC2, which was slightly increased if constructs with an intron and a hemagglutinin (HA) tag in addition to P19 expression were used. AbMV AC2 properties are discussed with reference to those of other geminiviruses with respect to charge, modification, and size in order to delimit possible reasons for the different behaviors.

IMPORTANCE
The (A)C2 genes encode a key pathogenicity factor of begomoviruses and curtoviruses in the plant virus family
Geminiviridae
. This factor has been implicated in the resistance breaking observed in agricultural cotton production. AC2 is a multifunctional protein involved in transcriptional control, gene silencing, and regulation of basal biosynthesis. Here, a new function of
Abutilon
mosaic virus AC2 in replication control is added as a feature of this protein in viral multiplication, providing a novel finding on geminiviral molecular biology.