bluuebunny/crossref_metadata_2025 huggingface dataset:
Ultimate Permeation Across Atomically Thin Porous Graphene
Authors: Celebi Kemal , Buchheim Jakob , Wyss Roman M. , Droudian Amirhossein , Gasser Patrick , Shorubalko Ivan , Kye Jeong-Il , Lee Changho , Park Hyung Gyu
Published: April 2014

Thin and Selective Outpourings

When using a membrane to separate materials, the efficiency of the separation is limited by how fast the gas or liquid passes through the membrane and by how selective it is. Thinner membranes usually allow for faster flow rates but are usually less selective. Attempting to maintain selectivity,

Celebi
et al.

(p.
289
) developed a sophisticated way to drill holes of controlled diameter in a graphene sheet about two layers thick. For such a thin membrane, the primary barriers to separation come from entrance and exit from the holes and not from the motion through the membrane.
The STAT3-Binding Long Noncoding RNA lnc-DC Controls Human Dendritic Cell Differentiation
Authors: Wang Pin , Xue Yiquan , Han Yanmei , Lin Li , Wu Cong , Xu Sheng , Jiang Zhengping , Xu Junfang , Liu Qiuyan , Cao Xuetao
Published: April 2014

L[i]nc to Dendritic Cell Activation

Long noncoding RNAs (lncRNAs) are important regulators of gene expression, but whether they are important regulators of the immune system is poorly understood.

Wang
et al.

(p.
310
) identify a lncRNA expressed exclusively in human dendritic cells (DC), called lnc-DC, that is required for optimal DC differentiation from human monocytes and that regulates DC activation of T cells. Lnc-DC interacts with the transcription factor STAT3, which is also required for DC development and function, to prevent interaction with and to block dephosphorylation by tyrosine phosphatase SHP1.
Myelin—More than Insulation
Authors: Fields R. Douglas
Published: April 2014

Unusual features of myelin in the mammalian cerebral cortex permit more complex forms of network integration.

[Also see Report by

Tomassy
et al.

]
Influenza and the Live Poultry Trade
Authors: Gao George F.
Published: April 2014

Live poultry trade at local markets has long been a part of China's national identity. From small villages to big cities, the gathering and selling of different birds in this vibrant atmosphere is at the heart of the country's cuisine culture. Unfortunately, the backdrop to this tradition has changed. Last year, the H7N9 virus, a new strain of influenza A, jumped from birds to humans, causing 144 cases of human infection and 47 deaths in China. Now a second wave of this flu is coursing through the country, with 258 confirmed cases and 99 deaths as of 8 April 2014. Scientific evidence points to a connection between the conditions at these live markets and the spread of flu, suggesting that until other means are found to prevent the transmission of or effectively treat the illness, China must shut down live poultry markets to prevent further spread of the virus and a possible global pandemic.
U.S. Support for ITER Wavers as Costs Spiral
Authors: Cho Adrian
Published: April 2014

Soaring cost estimates are jeopardizing the U.S. contribution to ITER, the massive international fusion energy project.
Almost-Earth Tantalizes Astronomers With Promise of Worlds to Come
Authors: Bhattacharjee Yudhijit
Published: April 2014

A new find from NASA's Kepler orbiting observatory is the first Earth-sized planet to be detected in the habitable zone of a star.
Into the Maelstrom
Authors: Kintisch Eli
Published: April 2014

Jennifer Francis has made waves linking the melting Arctic to extreme weather around the world. But a storm of criticism has forced the climate scientist to defend her hypothesis.
Safety and Immunogenicity of a Vero Cell Culture-Derived Whole-Virus H5N1 Influenza Vaccine in Chronically Ill and Immunocompromised Patients
Authors: van der Velden Maikel V. W. , Geisberger Alexander , Dvorak Thomas , Portsmouth Daniel , Fritz Richard , Crowe Brian A. , Herr Wolfgang , Distler Eva , Wagner Eva M. , Zeitlinger Markus , Sauermann Robert , Stephan Christoph , Ehrlich Hartmut J. , Barrett P. Noel , Aichinger Gerald
Published: April 2014

The development of vaccines against H5N1 influenza A viruses is a cornerstone of pandemic preparedness. Clinical trials of H5N1 vaccines have been undertaken in healthy subjects, but studies in risk groups have been lacking. In this study, the immunogenicity and safety of a nonadjuvanted cell culture-derived whole-virus H5N1 vaccine were assessed in chronically ill and immunocompromised adults. Subjects received two priming immunizations with a clade 1 A/Vietnam H5N1 influenza vaccine, and a subset also received a booster immunization with a clade 2.1 A/Indonesia H5N1 vaccine 12 to 24 months later. The antibody responses in the two populations were assessed by virus neutralization and single radial hemolysis assays. The T-cell responses in a subset of immunocompromised patients were assessed by enzyme-linked immunosorbent spot assay (ELISPOT). The priming and the booster vaccinations were safe and well tolerated in the two risk populations, and adverse reactions were predominantly mild and transient. The priming immunizations induced neutralizing antibody titers of ≥1:20 against the A/Vietnam strain in 64.2% of the chronically ill and 41.5% of the immunocompromised subjects. After the booster vaccination, neutralizing antibody titers of ≥1:20 against the A/Vietnam and A/Indonesia strains were achieved in 77.5% and 70.8%, respectively, of chronically ill subjects and in 71.6% and 67.5%, respectively, of immunocompromised subjects. The T-cell responses against the two H5N1 strains increased significantly over the baseline values. Substantial heterosubtypic T-cell responses were elicited against the 2009 pandemic H1N1 virus and seasonal A(H1N1), A(H3N2), and B subtypes. There was a significant correlation between T-cell responses and neutralizing antibody titers. These data indicate that nonadjuvanted whole-virus cell culture-derived H5N1 influenza vaccines are suitable for immunizing chronically ill and immunocompromised populations. (This study is registered at ClinicalTrials.gov under registration no. NCT00711295.)
Serological Diagnostic Assays for HIV-Associated Tuberculosis in Sub-Saharan Africa?
Authors: Lawn Stephen D.
Published: April 2014

In this issue of
Clinical and Vaccine Immunology
, Siev and colleagues present an evaluation of antibody responses to four immunodominant proteins of
Mycobacterium tuberculosis
in patients with HIV-associated pulmonary tuberculosis (TB) in South Africa (M. Siev, D. Wilson, S. Kainth, V. O. Kasprowicz, C. M. Feintuch, E. Jenny-Avital, and J. J. Achkar, 21:791–798, 2014, doi:
http://dx.doi.org/10.1128/CVI.00805-13
). This commentary discusses the enormous need for simple point-of-care assays for tuberculosis (TB) diagnosis in patients with and without HIV coinfection in high-burden settings and considers the potential role of serological assays and the huge challenges inherent in developing and validating such assays.
Validation of an Oligonucleotide Ligation Assay for Quantification of Human Immunodeficiency Virus Type 1 Drug-Resistant Mutants by Use of Massively Parallel Sequencing
Authors: Beck Ingrid A. , Deng Wenjie , Payant Rachel , Hall Robert , Bumgarner Roger E. , Mullins James I. , Frenkel Lisa M.
Published: April 2014

Global HIV treatment programs need sensitive and affordable tests to monitor HIV drug resistance. We compared mutant detection by the oligonucleotide ligation assay (OLA), an economical and simple test, to massively parallel sequencing. Nonnucleoside reverse transcriptase inhibitor (K103N, V106M, Y181C, and G190A) and lamivudine (M184V) resistance mutations were quantified in blood-derived plasma RNA and cell DNA specimens by OLA and 454 pyrosequencing. A median of 1,000 HIV DNA or RNA templates (range, 163 to 1,874 templates) from blood specimens collected in Mozambique (
n
= 60) and Kenya (
n
= 51) were analyzed at 4 codons in each sample (
n
= 441 codons assessed). Mutations were detected at 75 (17%) codons by OLA sensitive to 2.0%, at 71 codons (16%;
P
= 0.78) by pyrosequencing using a cutoff value of ≥2.0%, and at 125 codons (28%;
P
< 0.0001) by pyrosequencing sensitive to 0.1%. Discrepancies between the assays included 15 codons with mutant concentrations of ∼2%, one at 8.8% by pyrosequencing and not detected by OLA, and one at 69% by OLA and not detected by pyrosequencing. The latter two cases were associated with genetic polymorphisms in the regions critical for ligation of the OLA probes and pyrosequencing primers, respectively. Overall, mutant concentrations quantified by the two methods correlated well across the codons tested (
R
2
> 0.8). Repeat pyrosequencing of 13 specimens showed reproducible detection of 5/24 mutations at <2% and 6/6 at ≥2%. In conclusion, the OLA and pyrosequencing performed similarly in the quantification of nonnucleoside reverse transcriptase inhibitor and lamivudine mutations present at >2% of the viral population in clinical specimens. While pyrosequencing was more sensitive, detection of mutants below 2% was not reproducible.
Detection of Neisseria meningitidis from Negative Blood Cultures and Cerebrospinal Fluid with the FilmArray Blood Culture Identification Panel
Authors: Pardo Joe , Klinker Kenneth P. , Borgert Samuel J. , Butler Brittany M. , Rand Kenneth H. , Iovine Nicole M.
Published: April 2014

The FilmArray blood culture identification (BCID) panel is a rapid molecular diagnostic test approved for use with positive blood culture material. We describe a fatal case of meningococcemia with central nervous system (CNS) involvement detected using the BCID test with culture-negative blood and cerebrospinal fluid.
High Applicability of a Novel Method for gp60-Based Subtyping of Cryptosporidium meleagridis
Authors: Stensvold Christen Rune , Beser Jessica , Axén Charlotte , Lebbad Marianne
Published: April 2014

Cryptosporidium meleagridisis a common cause of cryptosporidiosis in avian hosts and the third most common species involved in human cryptosporidiosis. Sequencing of the highly polymorphic 60-kDa glycoprotein (gp60) gene is a frequently used tool for investigation of the genetic diversity and transmission dynamics ofCryptosporidium. However, few studies have included gp60 sequencing ofC. meleagridis. One explanation may be that the gp60 primers currently in use are based onCryptosporidium hominisandCryptosporidium parvumsequence data, potentially limiting successful amplification of theC. meleagridisgp60 gene. We therefore aimed to design primers for better gp60 subtyping ofC. meleagridis. Initially, ∼1,440 bp of the gp60 locus of sevenC. meleagridisisolates were amplified using primers flanking the open reading frame. The obtained sequence data (∼1,250 bp) were used to design primers for a nested PCR targetingC. meleagridis. Twenty isolates (16 from human and 4 from poultry) previously identified asC. meleagridisby analysis of small subunit (SSU) rRNA genes were investigated. Amplicons of the expected size (∼900 bp) were obtained from all 20 isolates. The subsequent sequence analysis identified 3 subtype families and 10 different subtypes. The most common subtype family, IIIb, was identified in 12 isolates, represented by 6 subtypes, 4 new and 2 previously reported. Subtype family IIIe was found in 3 isolates represented by 3 novel, distinct subtypes. Finally, IIIgA31G3R1 was found in 1 human isolate and 4 poultry isolates, all originating from a previously reportedC. meleagridisoutbreak at a Swedish organic farm.
Determining the Optimal Ceftriaxone MIC for Triggering Extended-Spectrum β-Lactamase Confirmatory Testing
Authors: Huang Yanjie , Carroll Karen C. , Cosgrove Sara E. , Tamma Pranita D.
Published: April 2014

As routine testing of clinical isolates for extended-spectrum β-lactamase (ESBL) production (screen plus phenotypic confirmatory testing) is no longer required by the Clinical and Laboratory Standards Institute (CLSI), a number of clinical microbiology laboratories use ceftriaxone MICs as a proxy means of identifying bacteria as potential ESBL producers. Data from 1,386 clinical isolates suggest that a ceftriaxone MIC cutoff of 8 μg/ml is an excellent predictor of ESBL production, with a positive predictive value and negative predictive value approaching 100% and 99.5%, respectively.
Inhibition Controls for Qualitative Real-Time PCR Assays: Are They Necessary for All Specimen Matrices?
Authors: Buckwalter S. P. , Sloan L. M. , Cunningham S. A. , Espy M. J. , Uhl J. R. , Jones M. F. , Vetter E. A. , Mandrekar J. , Cockerill F. R. , Pritt B. S. , Patel R. , Wengenack N. L.
Published: April 2014

A retrospective analysis of 386,706 specimens representing a variety of matrix types used in qualitative real-time PCR assays determined the overall inhibition rate to be 0.87% when the inhibition control was added preextraction to 5,613 specimens and 0.01% when the inhibition control was added postextraction but preamplification in 381,093 specimens. Inhibition rates of ≤1% were found for all specimen matrix types except urine and formalin-fixed, paraffin-embedded tissue.
Workflow and Maintenance Characteristics of Five Automated Laboratory Instruments for the Diagnosis of Sexually Transmitted Infections
Authors: Ratnam Sam , Jang Dan , Gilchrist Jodi , Smieja Marek , Poirier Andre , Hatchette Todd , Flandin Jean-Frederic , Chernesky Max
Published: April 2014

The choice of a suitable automated system for a diagnostic laboratory depends on various factors. Comparative workflow studies provide quantifiable and objective metrics to determine hands-on time during specimen handling and processing, reagent preparation, return visits and maintenance, and test turnaround time and throughput. Using objective time study techniques, workflow characteristics for processing 96 and 192 tests were determined on m2000 RealTi
m
e (Abbott Molecular), Viper XTR (Becton Dickinson), cobas 4800 (Roche Molecular Diagnostics), Tigris (Hologic Gen-Probe), and Panther (Hologic Gen-Probe) platforms using second-generation assays for
Chlamydia trachomatis
and
Neisseria gonorrhoeae
. A combination of operational and maintenance steps requiring manual labor showed that Panther had the shortest overall hands-on times and Viper XTR the longest. Both Panther and Tigris showed greater efficiency whether 96 or 192 tests were processed. Viper XTR and Panther had the shortest times to results and m2000 RealTi
m
e the longest. Sample preparation and loading time was the shortest for Panther and longest for cobas 4800. Mandatory return visits were required only for m2000 RealTi
m
e and cobas 4800 when 96 tests were processed, and both required substantially more hands-on time than the other systems due to increased numbers of return visits when 192 tests were processed. These results show that there are substantial differences in the amount of labor required to operate each system. Assay performance, instrumentation, testing capacity, workflow, maintenance, and reagent costs should be considered in choosing a system.
Use and Limits of (1-3)-β-
d
-Glucan Assay (Fungitell), Compared to Galactomannan Determination (Platelia Aspergillus), for Diagnosis of Invasive Aspergillosis
Authors: Sulahian Annie , Porcher Raphael , Bergeron Anne , Touratier Sophie , Raffoux Emmanuel , Menotti Jean , Derouin Francis , Ribaud Patricia
Published: April 2014

This study was undertaken to examine the performance of the Fungitell β-glucan (BG) assay, to compare it with that of the galactomannan (GM) test for the diagnosis of invasive aspergillosis (IA) in patients with hematological malignancies, and to examine the rates of false-positive BG and GM test results due to β-lactam antibiotics among sera of patients with Gram-positive or Gram-negative bacteremia and selected sera with false-positive results from the GM test. Serum samples from 105 patients with proven (
n
= 14) or probable (
n
= 91) IA, 97 hematology patients at risk for invasive fungal infections, 50 healthy blood donors, and 60 patients with bacteremia were used to study the sensitivities and specificities of the assays. The GM test was more specific than the BG assay (97% versus 82%, respectively;
P
= 0.0001) and the BG assay was more sensitive than the GM test (81% versus 49%, respectively;
P
< 0.0001) for IA diagnosis. The study of 49 separate batches of β-lactam antibiotics showed high and very similar rates of false-positive results for the GM and BG assays (29 and 33%, respectively;
P
= 0.82) but with an almost complete lack of concordance between the 2 assays. For patients with bacteremia, the rate of false-positive results was much higher with the BG test than with the GM test (37% versus 2%, respectively;
P
< 0.0001), with no significant difference between Gram-positive and Gram-negative bacteremia. In conclusion, the BG test may be useful for the diagnosis of IA because of its high sensitivity in comparison with the GM test, but the overall benefit of this assay remains limited because of its inadequate specificity and its cost.
Comparison of Real-Time PCR and Antigen Assays for Detection of Hepatitis E Virus in Blood Donors
Authors: Vollmer T. , Knabbe C. , Dreier J.
Published: April 2014

Hepatitis E virus (HEV) infection is recognized as an emerging and often undiagnosed disease in industrialized countries, with asymptomatic infections actually occurring in blood donors. Sensitive detection of HEV-RNA is crucial for diagnosis and monitoring of disease progression. We evaluated the analytical sensitivity and performance of three HEV RT-PCR assays (RealStar HEV reverse transcription-PCR [RT-PCR], hepatitis@ceeramTools, and ampliCube HEV RT-PCR) for screening of individuals for HEV infections (ID-nucleic acid amplification technology [ID-NAT]) and for blood donor pool screening (minipool-NAT [MP-NAT]). RNA was extracted using NucliSens easyMAG (ID-NAT) and a high-volume extraction protocol (4.8 ml, chemagic Viral 5K, MP-NAT). Three NAT assays were evaluated for ID-NAT but only two assays for MP-NAT due to inhibition of the ampliCube HEV RT-PCR kit using the corresponding RNA extract. Assays provided good analytical sensitivity, ranging from 37.8 to 180.1 IU/ml (ID-NAT) and from 4.7 to 91.2 IU/ml (MP-NAT). The applicability of HEV antigen (HEV-Ag) screening was compared to that of RT-PCR screening and detection of HEV-IgM antibodies using seroconversion panels of 10 HEV genotype 3-infected individuals. Four individuals revealed a positive HEV-Ag detection result, with corresponding viremias ranging from 1.92E + 03 to 2.19E + 05 IU/ml, while the progression of HEV-Ag followed that of HEV viremia. The other six individuals showed no presence of HEV-Ag although the corresponding viremias were also in the range of >1.0E + 03. Anti-HEV-IgM antibodies were detectable in seven donors; one donor presented parallel positivities of HEV-Ag and anti-HEV IgM. The evaluated NAT methods present powerful tools providing sensitive HEV detection. Application of HEV-Ag or anti-HEV IgM screening is currently inferior for the early detection of HEV infection due to the decreased sensitivity compared to NAT methods.
Interspecies Differences in Virus Uptake versus Cardiac Function of the Coxsackievirus and Adenovirus Receptor
Authors: Freiberg Fabian , Sauter Martina , Pinkert Sandra , Govindarajan Thirupugal , Kaldrack Joanna , Thakkar Meghna , Fechner Henry , Klingel Karin , Gotthardt Michael
Published: April 2014

The coxsackievirus and adenovirus receptor (CAR) is a cell contact protein with an important role in virus uptake. Its extracellular immunoglobulin domains mediate the binding to coxsackievirus and adenovirus as well as homophilic and heterophilic interactions between cells. The cytoplasmic tail links CAR to the cytoskeleton and intracellular signaling cascades. In the heart, CAR is crucial for embryonic development, electrophysiology, and coxsackievirus B infection. Noncardiac functions are less well understood, in part due to the lack of suitable animal models. Here, we generated a transgenic mouse that rescued the otherwise embryonic-lethal CAR knockout (KO) phenotype by expressing chicken CAR exclusively in the heart. Using this rescue model, we addressed interspecies differences in coxsackievirus uptake and noncardiac functions of CAR. Survival of the noncardiac CAR KO (ncKO) mouse indicates an essential role for CAR in the developing heart but not in other tissues. In adult animals, cardiac activity was normal, suggesting that chicken CAR can replace the physiological functions of mouse CAR in the cardiomyocyte. However, chicken CAR did not mediate virus entry
in vivo
, so that hearts expressing chicken instead of mouse CAR were protected from infection and myocarditis. Comparison of sequence homology and modeling of the D1 domain indicate differences between mammalian and chicken CAR that relate to the sites important for virus binding but not those involved in homodimerization. Thus, CAR-directed anticoxsackievirus therapy with only minor adverse effects in noncardiac tissue could be further improved by selectively targeting the virus-host interaction while maintaining cardiac function.

IMPORTANCE
Coxsackievirus B3 (CVB3) is one of the most common human pathogens causing myocarditis. Its receptor, the coxsackievirus and adenovirus receptor (CAR), not only mediates virus uptake but also relates to cytoskeletal organization and intracellular signaling. Animals without CAR die prenatally with major cardiac malformations. In the adult heart, CAR is important for virus entry and electrical conduction, but its nonmuscle functions are largely unknown. Here, we show that chicken CAR expression exclusively in the heart can rescue the otherwise embryonic-lethal CAR knockout phenotype but does not support CVB3 infection of adult cardiomyocytes. Our findings have implications for the evolution of virus-host versus physiological interactions involving CAR and could help to improve future coxsackievirus-directed therapies inhibiting virus replication while maintaining CAR's cellular functions.
Evolution of Puma Lentivirus in Bobcats (Lynx rufus) and Mountain Lions (Puma concolor) in North America
Authors: Lee Justin S. , Bevins Sarah N. , Serieys Laurel E. K. , Vickers Winston , Logan Ken A. , Aldredge Mat , Boydston Erin E. , Lyren Lisa M. , McBride Roy , Roelke-Parker Melody , Pecon-Slattery Jill , Troyer Jennifer L. , Riley Seth P. , Boyce Walter M. , Crooks Kevin R. , VandeWoude Sue
Published: April 2014

Mountain lions (
Puma concolor
) throughout North and South America are infected with puma lentivirus clade B (PLVB). A second, highly divergent lentiviral clade, PLVA, infects mountain lions in southern California and Florida. Bobcats (
Lynx rufus
) in these two geographic regions are also infected with PLVA, and to date, this is the only strain of lentivirus identified in bobcats. We sequenced full-length PLV genomes in order to characterize the molecular evolution of PLV in bobcats and mountain lions. Low sequence homology (88% average pairwise identity) and frequent recombination (1 recombination breakpoint per 3 isolates analyzed) were observed in both clades. Viral proteins have markedly different patterns of evolution; sequence homology and negative selection were highest in Gag and Pol and lowest in Vif and Env. A total of 1.7% of sites across the PLV genome evolve under positive selection, indicating that host-imposed selection pressure is an important force shaping PLV evolution. PLVA strains are highly spatially structured, reflecting the population dynamics of their primary host, the bobcat. In contrast, the phylogeography of PLVB reflects the highly mobile mountain lion, with diverse PLVB isolates cocirculating in some areas and genetically related viruses being present in populations separated by thousands of kilometers. We conclude that PLVA and PLVB are two different viral species with distinct feline hosts and evolutionary histories.

IMPORTANCE
An understanding of viral evolution in natural host populations is a fundamental goal of virology, molecular biology, and disease ecology. Here we provide a detailed analysis of puma lentivirus (PLV) evolution in two natural carnivore hosts, the bobcat and mountain lion. Our results illustrate that PLV evolution is a dynamic process that results from high rates of viral mutation/recombination and host-imposed selection pressure.
Analysis of Lujo Virus Cell Entry using Pseudotype Vesicular Stomatitis Virus
Authors: Tani Hideki , Iha Koichiro , Shimojima Masayuki , Fukushi Shuetsu , Taniguchi Satoshi , Yoshikawa Tomoki , Kawaoka Yoshihiro , Nakasone Naoe , Ninomiya Haruaki , Saijo Masayuki , Morikawa Shigeru
Published: April 2014

Several arenaviruses are known to cause viral hemorrhagic fever (VHF) in sub-Saharan Africa and South America, where VHF is a major public health and medical concern. The biosafety level 4 categorization of these arenaviruses restricts their use and has impeded biological studies, including therapeutic drug and/or vaccine development. Due to difficulties associated with handling live viruses, pseudotype viruses, which transiently bear arenavirus envelope proteins based on vesicular stomatitis virus (VSV) or retrovirus, have been developed as surrogate virus systems. Here, we report the development of a pseudotype VSV bearing each envelope protein of various species of arenaviruses (AREpv), including the newly identified Lujo virus (LUJV) and Chapare virus. Pseudotype arenaviruses generated in 293T cells exhibited high infectivity in various mammalian cell lines. The infections by New World and Old World AREpv were dependent on their receptors (human transferrin receptor 1 [hTfR1] and α-dystroglycan [αDG], respectively). However, infection by pseudotype VSV bearing the LUJV envelope protein (LUJpv) occurred independently of hTfR1 and αDG, indicating that LUJpv utilizes an unidentified receptor. The pH-dependent endocytosis of AREpv was confirmed by the use of lysosomotropic agents. The fusion of cells expressing these envelope proteins, except for those expressing the LUJV envelope protein, was induced by transient treatment at low pH values. LUJpv infectivity was inhibited by U18666A, a cholesterol transport inhibitor. Furthermore, the infectivity of LUJpv was significantly decreased in the Niemann-Pick C1 (NPC1)-deficient cell line, suggesting the necessity for NPC1 activity for efficient LUJpv infection.

IMPORTANCE
LUJV is a newly identified arenavirus associated with a VHF outbreak in southern Africa. Although cell entry for many arenaviruses has been studied, cell entry for LUJV has not been characterized. In this study, we found that LUJpv utilizes neither αDG nor hTfR1 as a receptor and found unique characteristics of LUJV glycoprotein in membrane fusion and cell entry. Proper exclusion of cholesterol or some kinds of lipids may play important roles in LUJpv cell entry.
Kaposi's Sarcoma-Associated Herpesvirus-Encoded LANA Interacts with Host KAP1 To Facilitate Establishment of Viral Latency
Authors: Sun Rui , Liang Deguang , Gao Yuan , Lan Ke
Published: April 2014

Kaposi's sarcoma-associated herpesvirus (KSHV) typically displays two different phases in its life cycle, the default latent phase and the lytic phase. There is a short period of lytic gene expression in the early stage of KSHV primary infection. The factors involved in the shutdown process of lytic gene expression are poorly identified. It has been shown that the latency-associated nuclear antigen (LANA) encoded by KSHV plays an important role in the establishment of viral latency. In screening, we identified a host protein, Krüppel-associated box domain-associated protein 1 (KAP1), that bound to LANA. We validated the interaction between LANA and KAP1
in vivo
and
in vitro
, as well as their colocalization in the nucleus. We mapped out that LANA interacted with both the N- and C-terminal domains of KAP1. Based on the interface of LANA-KAP1 interaction determined, we proved that LANA recruited KAP1 to the RTA promoter region of the KSHV genome. We revealed that KAP1 was involved in transcriptional repression by LANA. We found multiple cooccupation sites of LANA and KAP1 on the whole KSHV genome by chromatin immunoprecipitation for sequencing (ChIP-seq) and demonstrated that LANA-recruited KAP1 played a critical role in the shutdown of lytic gene expression during the early stage of KSHV primary infection. Taken together, our data suggest that LANA interacts with KAP1 and represses lytic gene expression to facilitate the establishment of KSHV latency.

IMPORTANCE
Our study revealed the mechanism of transcriptional repression by LANA during KSHV primary infection, providing new insights into the process of KSHV latency establishment.
Anti-Inflammatory Cytokines Directly Inhibit Innate but Not Adaptive CD8+T Cell Functions
Authors: Freeman Bailey E. , Meyer Christine , Slifka Mark K.
Published: April 2014

Virus-specific CD8+T cells provide classical adaptive immunity by responding to cognate peptide antigen, but they may also act in an “innate” capacity by responding directly to cytokine stimulation. Here, we examined regulation of these distinct T cell functions by anti-inflammatory cytokines (interleukin-4 [IL-4], IL-10, and transforming growth factor β [TGF-β]). Innate gamma interferon (IFN-γ) production by CD8+T cells following exposure to IL-12 plus IL-18, IL-12 plus tumor necrosis factor alpha (TNF-α), or IL-12 plus IL-15 was inhibited by exposure to anti-inflammatory cytokines either before or shortly after stimulation. However, inhibition was not universal, as other activation parameters, including upregulation of CD25 and CD69, remained largely unaltered. In contrast, peptide-specific T cell responses were resistant to inhibition by anti-inflammatory cytokines. This was not due to downregulation of cytokine receptor expression or an inability to signal through cytokine receptors since phosphorylation of STAT proteins remained intact. These results highlight key differences in cytokine-mediated regulation of innate and adaptive T cell functions, which may help balance effective antiviral immune responses while reducing T cell-mediated immunopathology.

IMPORTANCEThis study demonstrates key differences between the regulation of “innate” and “adaptive” CD8+T cell functions following activation by innate cytokines or viral peptide. Innate production of IFN-γ by CD8+T cells following exposure to IL-12 plus IL-18, IL-12 plus TNF-α, or IL-12 plus IL-15 was inhibited by exposure to anti-inflammatory cytokines (IL-4, IL-10, and TGF-β). However, inhibition was not universal, as other activation parameters, including upregulation of CD25 and CD69, remained largely unaltered. In contrast, peptide-specific T cell responses were resistant to inhibition by anti-inflammatory cytokines. This distinct regulation of innate and adaptive T cell functions may serve to reduce T cell-mediated immunopathology while still allowing for effective antiviral responses at a site of infection.
Intrahost Dynamics of Influenza Virus Reassortment
Authors: Tao Hui , Steel John , Lowen Anice C.
Published: April 2014

The segmented nature of the influenza virus genome allows reassortment between coinfecting viruses. This process of genetic exchange vastly increases the diversity of circulating influenza viruses. The importance of reassortment to public health is clear from its role in the emergence of a number of epidemiologically important viruses, including novel pandemic and epidemic strains. To gauge its impact on within-host genomic variation, we tracked reassortment in coinfected guinea pigs over time and given matched or discordant doses of coinfecting viruses. To ensure unbiased detection of reassortants, we used parental viruses of equivalent fitness that differ only by noncoding nucleotide changes. These viruses were based on the isolate A/Panama/2007/1999 (H3N2). At a dose of 2 × 10
2
PFU, one parental virus was absent from each guinea pig throughout the time course, indicating the presence of a bottleneck. With an intermediate dose of 2 × 10
3
PFU, genomic diversity present in nasal lavage samples increased from 1 to 3 days postinfection (dpi) and then declined by 6 dpi. With a high dose of 2 × 10
6
PFU, however, reassortment levels were high (avg. 59%) at 1 dpi and remained stable. Even late in the course of infection, parental viruses were not eclipsed by reassortants, suggesting that a uniformly high multiplicity of infection was not achieved
in vivo
. Inoculation with ∼10-fold discordant doses did not reduce reassortment relative to equivalent inputs but markedly changed the spectrum of genotypes produced. Our data reveal the potential for reassortment to contribute to intrahost diversity in mixed influenza virus infection.

IMPORTANCE
Influenza virus reassortment is prevalent in nature and is a major contributor to the diversity of influenza viruses circulating in avian, swine, human and other host species. This diversity, in turn, increases the potential for influenza viruses to evade selective pressures or adapt to new host environments. As examples, reassortment was key to the emergence of the 1957, 1968, and 2009 pandemics; the unusually severe influenza epidemics of 2003, 1951, and 1947; and the rise in adamantane resistance among currently circulating human H3N2 viruses. We reveal here the diversity of viral genotypes generated over time in a host coinfected with two influenza viruses. We found that intrahost diversity driven by reassortment is dynamic and dependent on the amount of each virus initiating infection. Our results demonstrate the readiness with which reassortant influenza viruses arise, offering new insight into this important mechanism of influenza virus evolution.
Induction of Gag-Specific CD4 T Cell Responses during Acute HIV Infection Is Associated with Improved Viral Control
Authors: Schieffer Miriam , Jessen Heiko K. , Oster Alexander F. , Pissani Franco , Soghoian Damien Z. , Lu Richard , Jessen Arne B. , Zedlack Carmen , Schultz Bruce T. , Davis Isaiah , Ranasinghe Srinika , Rosenberg Eric S. , Alter Galit , Schumann Ralf R. , Streeck Hendrik
Published: April 2014

Effector CD4 T cell responses have been shown to be critically involved in the containment and clearance of viral pathogens. However, their involvement in the pathogenesis of HIV infection is less clear, given their additional role as preferred viral targets. We previously demonstrated that the presence of HIV-specific CD4 T cell responses is somewhat associated with HIV control and that specific CD4 T cell functions, such as direct cytolytic activity, can contribute to control of HIV viremia. However, little is known about how the induction of HIV-specific CD4 T cell responses during acute HIV infection influences disease progression and whether responses induced during the early phase of infection are preferentially depleted. We therefore longitudinally assessed, in a cohort of 55 acutely HIV-infected individuals, HIV-specific CD4 T cell responses from acute to chronic infection. Interestingly, we found that the breadth, magnitude, and protein dominance of HIV-specific CD4 T cell responses remained remarkably stable over time. Moreover, we found that the epitopes targeted at a high frequency in acute HIV infection were recognized at the same frequency by HIV-specific CD4 T cells in chronic HIV infection. Interestingly the induction of Gag-specific CD4 T cell responses in acute HIV infection was significantly inversely correlated with viral set point in chronic HIV infection (
R
= −0.5;
P
= 0.03), while the cumulative contribution of Env-specific CD4 T cell responses showed the reverse effect. Moreover, individuals with HIV-specific CD4 T cell responses dominantly targeting Gag over Env in acute HIV infection remained off antiretroviral therapy significantly longer (
P
= 0.03; log rank). Thus, our data suggest that the induction of HIV-specific CD4 T cell responses during acute HIV infection is beneficial overall and does not fuel disease progression.

IMPORTANCE
CD4 T cells are critical for the clearance and control of viral infections. However, HIV preferentially infects HIV-specific CD4 T cells. Thus, their contribution to the control of HIV viremia is uncertain. Here, we study HIV-specific CD4 T cell responses from acute to chronic HIV infection and show that the generation of certain CD4 responses is associated with control rather than disease progression.
Characterization of the Nucleocytoplasmic Shuttle of the Matrix Protein of Influenza B Virus
Authors: Cao Shuai , Jiang Jingwen , Li Jing , Li Yan , Yang Limin , Wang Shanshan , Yan Jinghua , Gao George F. , Liu Wenjun
Published: April 2014

Influenza B virus is an enveloped negative-strand RNA virus that contributes considerably to annual influenza illnesses in human. The matrix protein of influenza B virus (BM1) acts as a cytoplasmic-nuclear shuttling protein during the early and late stages of infection. The mechanism of this intracellular transport of BM1 was revealed through the identification of two leucine-rich CRM1-dependent nuclear export signals (NESs) (3 to 14 amino acids [aa] and 124 to 133 aa), one bipartite nuclear localization signal (NLS) (76 to 94 aa), and two phosphorylation sites (80T and 84S) in BM1. The biological function of the NLS and NES regions were determined through the observation of the intracellular distribution of enhanced green fluorescent protein (EGFP)-tagged signal peptides, and wild-type, NES-mutant, and NLS-mutant EGFP-BM1. Furthermore, the NLS phosphorylation sites 80T and 84S, were found to be required for the nuclear accumulation of EGFP-NLS and for the efficient binding of EGFP-BM1 to human importin-α1. Moreover, all of these regions/sites were required for the generation of viable influenza B virus in a 12-plasmid virus rescue system.

IMPORTANCE
This study expands our understanding of the life cycle of influenza B virus by defining the dynamic mechanism of the nucleocytoplasmic shuttle of BM1 and could provide a scientific basis for the development of antiviral medication.
Role of Herpes Simplex Virus 1 Immediate Early Protein ICP22 in Viral Nuclear Egress
Authors: Maruzuru Yuhei , Shindo Keiko , Liu Zhuoming , Oyama Masaaki , Kozuka-Hata Hiroko , Arii Jun , Kato Akihisa , Kawaguchi Yasushi
Published: April 2014

In order to investigate the novel function(s) of the herpes simplex virus 1 (HSV-1) immediate early protein ICP22, we screened for ICP22-binding proteins in HSV-1-infected cells. Our results were as follows. (i) Tandem affinity purification of ICP22 from infected cells, coupled with mass spectrometry-based proteomics and subsequent analyses, demonstrates that ICP22 forms a complex(es) with the HSV-1 proteins UL31, UL34, UL47 (or VP13/14), and/or Us3. All these proteins were previously reported to be important for viral egress through the nuclear membrane. (ii) ICP22 colocalizes with UL31 and UL34 at the nuclear membrane in wild-type HSV-1-infected cells. (iii) The UL31-null mutation prevents the targeting of ICP22 to the nuclear membrane. (iv) The ICP22-null mutation resulted in UL31 and UL34 being mislocalized in the endoplasmic reticulum (in addition to the nuclear membrane) and significantly reduced numbers of primary enveloped virions in the perinuclear space, although capsids accumulated in the nuclei. Collectively, these results suggest that (i) ICP22 interacts with HSV-1 regulators of nuclear egress, including UL31, UL34, UL47, and Us3 in HSV-1-infected cells; (ii) UL31 mediates the recruitment and anchorage of ICP22 at the nuclear membrane; and (iii) ICP22 plays a regulatory role in HSV-1 primary envelopment, probably by interacting with and regulating UL31 and UL34. Here we report a previously unknown function for ICP22 in the regulation of HSV-1 nuclear egress.

IMPORTANCE
The herpes simplex virus 1 (HSV-1) immediate early protein ICP22 is recognized primarily as a regulator of viral gene expression. In this study, we show that ICP22 interacts with the HSV-1 proteins UL31 and UL34, which play crucial roles at the nuclear membrane in HSV-1 primary envelopment during viral nuclear egress. We also demonstrate that UL31 is required for the targeting of ICP22 to the nuclear membrane and that ICP22 is required for the correct localization of UL31 and/or UL34. Furthermore, we confirm that ICP22 is required for efficient HSV-1 primary envelopment during viral nuclear egress. Thus, we report, for the first time, that ICP22 plays a regulatory role in HSV-1 nuclear egress.
Type I Interferon Signals in Macrophages and Dendritic Cells Control Dengue Virus Infection: Implications for a New Mouse Model To Test Dengue Vaccines
Authors: Züst Roland , Toh Ying-Xiu , Valdés Iris , Cerny Daniela , Heinrich Julia , Hermida Lisset , Marcos Ernesto , Guillén Gerardo , Kalinke Ulrich , Shi Pei-Yong , Fink Katja
Published: April 2014

Dengue virus (DENV) infects an estimated 400 million people every year, causing prolonged morbidity and sometimes mortality. Development of an effective vaccine has been hampered by the lack of appropriate small animal models; mice are naturally not susceptible to DENV and only become infected if highly immunocompromised. Mouse models lacking both type I and type II interferon (IFN) receptors (AG129 mice) or the type I IFN receptor (IFNAR
−/−
mice) are susceptible to infection with mouse-adapted DENV strains but are severely impaired in mounting functional immune responses to the virus and thus are of limited use for study. Here we used conditional deletion of the type I IFN receptor (IFNAR) on individual immune cell subtypes to generate a minimally manipulated mouse model that is susceptible to DENV while retaining global immune competence. Mice lacking IFNAR expression on CD11c
+
dendritic cells and LysM
+
macrophages succumbed completely to DENV infection, while mice deficient in the receptor on either CD11c
+
or LysM
+
cells were susceptible to infection but often resolved viremia and recovered fully from infection. Conditional IFNAR mice responded with a swift and strong CD8
+
T-cell response to viral infection, compared to a weak response in IFNAR
−/−
mice. Furthermore, mice lacking IFNAR on either CD11c
+
or LysM
+
cells were also sufficiently immunocompetent to raise a protective immune response to a candidate subunit vaccine against DENV-2. These data demonstrate that mice with conditional deficiencies in expression of the IFNAR represent improved models for the study of DENV immunology and screening of vaccine candidates.

IMPORTANCE
Dengue virus infects 400 million people every year worldwide, causing 100 million clinically apparent infections, which can be fatal if untreated. Despite many years of research, there are no effective vaccine and no antiviral treatment available for dengue. Development of vaccines has been hampered in particular by the lack of a suitable small animal model. Mouse models used to test dengue vaccine are deficient in interferon (IFN) type I signaling and severely immunocompromised and therefore likely not ideal for the testing of vaccines. In this study, we explored alternative models lacking the IFN receptor only on certain cell types. We show that mice lacking the IFN receptor on either CD11c- or LysM-expressing cells (conditional IFNAR mice) are susceptible to dengue virus infection. Importantly, we demonstrate that conditional IFN receptor knockout mice generate a better immune response to live virus and a candidate dengue vaccine compared to IFNAR mice and are resistant to subsequent challenge.
Bacillus subtilis
Systems Biology: Applications of -Omics Techniques to the Study of Endospore Formation
Authors: Bate Ashley R. , Bonneau Richard , Eichenberger Patrick
Published: April 2014

Endospore-forming bacteria, with
Bacillus subtilis
being the prevalent model organism, belong to the phylum Firmicutes. Although the last common ancestor of all
Firmicutes
is likely to have been an endospore-forming species, not every lineage in the phylum has maintained the ability to produce endospores (hereafter, spores). In 1997, the release of the full genome sequence for
B. subtilis
strain 168 marked the beginning of the genomic era for the study of spore formation (sporulation). In this original genome sequence, 139 of the 4,100 protein-coding genes were annotated as sporulation genes. By the time a revised genome sequence with updated annotations was published in 2009, that number had increased significantly, especially since transcriptional profiling studies (transcriptomics) led to the identification of several genes expressed under the control of known sporulation transcription factors. Over the past decade, genome sequences for multiple spore-forming species have been released (including several strains in the
Bacillus anthracis
/
Bacillus cereus
group and many
Clostridium
species), and phylogenomic analyses have revealed many conserved sporulation genes. Parallel advances in transcriptomics led to the identification of small untranslated regulatory RNAs (sRNAs), including some that are expressed during sporulation. An extended array of -omics techniques, i.e., techniques designed to probe gene function on a genome-wide scale, such as proteomics, metabolomics, and high-throughput protein localization studies, have been implemented in microbiology. Combined with the use of new computational methods for predicting gene function and inferring regulatory relationships on a global scale, these -omics approaches are uncovering novel information about sporulation and a variety of other bacterial cell processes.
Electroacupuncture at St36 Accelerates the Recovery of Gastrointestinal Motility after Colorectal Surgery: A Randomised Controlled Trial
Authors: Zhang Zhaodi , Wang Changsong , Li Quanyi , Zhang Mingyue , Zhao Haifang , Dong Linlin , Wang Guonian , Jin Yan
Published: April 2014

Objectives

To evaluate whether electroacupuncture (EA) at ST36 can accelerate the recovery of gastrointestinal motility after colorectal surgery.

Methods

Forty patients of American Society of Anesthesiologists physical status II and III undergoing elective open resection of malignant colorectal tumours were included in this study. Using a sealed envelope method, the patients were randomly divided into two groups either receiving EA (EA group) or sham EA (SEA group). Data regarding the recovery of bowel function (times to the first bowel sounds, passage of flatus and defaecation) were collected and analysed.

Results

In the EA group, the time intervals from surgery to the first bowel movement and passage of flatus were shorter than in the SEA group (13±10 h vs 19±13 h, p<0.05 and 23±14 h vs 32±18 h, p<0.05, respectively). There was no significant difference between the groups regarding the time to first defaecation (68±45 h vs 72±53 h, p>0.05).

Conclusions

EA at ST36 accelerates the recovery of gastrointestinal motility after colorectal surgery.

Trial Registration

JJ22011-15.
Solar urticaria in a 1-year-old infant: diagnosis and management
Authors: Hochstadter Elana Fay , Ben-Shoshan Moshe
Published: April 2014

Solar urticaria is a type of inducible urticaria triggered by sun exposure resulting in hives on exposed skin as well as risk of developing systemic symptoms. Most cases are reported in the adult population. However, cases do occur in children. We present a case of visible light induced urticaria in a 12-month-old girl that demonstrates the need for accurate diagnosis and appropriate management.
Paranasal sinus and retro-orbital metastasis in a case of breast carcinoma: a clinicoradiological review
Authors: Tiwari Vivek , Pande Subodh Chandra , Verma Kamal , Goel Sandeep
Published: April 2014

Metastatic spread from non-head and neck tumours (H&N) to the paranasal and orbital region is extremely rare. We present a case of breast cancer metastasis to the left sphenoid wing, lateral wall left orbit, lateral wall left sphenoid sinus, medial and lateral wall left maxillary sinus and left pterygoid plates. We provide an exhaustive clinicoradiological imaging with a brief literature review.
Lateral foot pain following open reduction and internal fixation of the fracture of the fifth metatarsal tubercle: treated by arthroscopic arthrolysis and endoscopic tenolysis
Authors: Lui Tun Hing
Published: April 2014

We report a case of fracture of the tuberosity of the fifth metatarsal which was managed by tension band wiring and bone grafting. It was complicated by symptomatic fibrosis of the operated site involving the cubometatarsal joint, peroneus brevis tendon, peroneus tertius tendon and the long extensor tendon to the fifth toe. This was successfully managed by arthroscopic lysis of the involved joint and tendons.