bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human MicroRNA miR-532-5p Exhibits Antiviral Activity against West Nile Virus via Suppression of Host Genes SESTD1 and TAB3 Required for Virus Replication
Authors: Slonchak Andrii , Shannon Rory P. , Pali Gabor , Khromykh Alexander A. 
Published: December 2015

ABSTRACT

West Nile virus (WNV) is a mosquito-transmitted flavivirus that naturally circulates between mosquitos and birds but can also infect humans, causing severe neurological disease. The early host response to WNV infection in vertebrates primarily relies on the type I interferon pathway; however, recent studies suggest that microRNAs (miRNAs) may also play a notable role. In this study, we assessed the role of host miRNAs in response to WNV infection in human cells. We employed small RNA sequencing (RNA-seq) analysis to determine changes in the expression of host miRNAs in HEK293 cells infected with an Australian strain of WNV, Kunjin (WNV
KUN
), and identified a number of host miRNAs differentially expressed in response to infection. Three of these miRNAs were confirmed to be significantly upregulated in infected cells by quantitative reverse transcription (qRT)-PCR and Northern blot analyses, and one of them, miR-532-5p, exhibited a significant antiviral effect against WNV
KUN
infection. We have demonstrated that miR-532-5p targets and downregulates expression of the host genes SESTD1 and TAB3 in human cells. Small interfering RNA (siRNA) depletion studies showed that both SESTD1 and TAB3 were required for efficient WNV
KUN
replication. We also demonstrated upregulation of mir-532-5p expression and a corresponding decrease in the expression of its targets, SESTD1 and TAB3, in the brains of WNV
KUN
-infected mice. Our results show that upregulation of miR-532-5p and subsequent suppression of the SESTD1 and TAB3 genes represent a host antiviral response aimed at limiting WNV
KUN
infection and highlight the important role of miRNAs in controlling RNA virus infections in mammalian hosts.

IMPORTANCE
West Nile virus (WNV) is a significant viral pathogen that poses a considerable threat to human health across the globe. There is no specific treatment or licensed vaccine available for WNV, and deeper insight into how the virus interacts with the host is required to facilitate their development. In this study, we addressed the role of host microRNAs (miRNAs) in antiviral response to WNV in human cells. We identified miR-532-5p as a novel antiviral miRNA and showed that it is upregulated in response to WNV infection and suppresses the expression of the host genes TAB3 and SESTD1 required for WNV replication. Our results show that upregulation of miR-532-5p and subsequent suppression of the SESTD1 and TAB3 genes represent an antiviral response aimed at limiting WNV infection and highlight the important role of miRNAs in controlling virus infections in mammalian hosts.
A Single Mutation at PB1 Residue 319 Dramatically Increases the Safety of PR8 Live Attenuated Influenza Vaccine in a Murine Model without Compromising Vaccine Efficacy
Authors: Cox Andrew , Dewhurst Stephen 
Published: December 2015

ABSTRACT
The live attenuated influenza vaccine (LAIV) is preferentially recommended for use in most children yet remains unsafe for the groups most at risk. Here we have improved the safety of a mouse-adapted live attenuated influenza vaccine containing the same attenuating amino acid mutations as in human LAIV by adding an additional mutation at PB1 residue 319. This results in a vaccine with a 20-fold decrease in protective efficacy and a 10,000-fold increase in safety.
Membrane Binding of the Rous Sarcoma Virus Gag Protein Is Cooperative and Dependent on the Spacer Peptide Assembly Domain
Authors: Dick Robert A. , Barros Marilia , Jin Danni , Lösche Mathias , Vogt Volker M. 
Published: December 2015

ABSTRACT

The principles underlying membrane binding and assembly of retroviral Gag proteins into a lattice are understood. However, little is known about how these processes are related. Using purified Rous sarcoma virus Gag and Gag truncations, we studied the interrelation of Gag-Gag interaction and Gag-membrane interaction. Both by liposome binding and by surface plasmon resonance on a supported bilayer, Gag bound to membranes much more tightly than did matrix (MA), the isolated membrane binding domain. In principle, this difference could be explained either by protein-protein interactions leading to cooperativity in membrane binding or by the simultaneous interaction of the N-terminal MA and the C-terminal nucleocapsid (NC) of Gag with the bilayer, since both are highly basic. However, we found that NC was not required for strong membrane binding. Instead, the spacer peptide assembly domain (SPA), a putative 24-residue helical sequence comprising the 12-residue SP segment of Gag and overlapping the capsid (CA) C terminus and the NC N terminus, was required. SPA is known to be critical for proper assembly of the immature Gag lattice. A single amino acid mutation in SPA that abrogates assembly
in vitro
dramatically reduced binding of Gag to liposomes.
In vivo
, plasma membrane localization was dependent on SPA. Disulfide cross-linking based on ectopic Cys residues showed that the contacts between Gag proteins on the membrane are similar to the known contacts in virus-like particles. Taken together, we interpret these results to mean that Gag membrane interaction is cooperative in that it depends on the ability of Gag to multimerize.

IMPORTANCE
The retroviral structural protein Gag has three major domains. The N-terminal MA domain interacts directly with the plasma membrane (PM) of cells. The central CA domain, together with immediately adjoining sequences, facilitates the assembly of thousands of Gag molecules into a lattice. The C-terminal NC domain interacts with the genome, resulting in packaging of viral RNA. For assembly
in vitro
with purified Gag, in the absence of membranes, binding of NC to nucleic acid somehow facilitates further Gag-Gag interactions that lead to formation of the Gag lattice. The contributions of MA-mediated membrane binding to virus particle assembly are not well understood. Here, we report that in the absence of nucleic acid, membranes provide a platform that facilitates Gag-Gag interactions. This study demonstrates that the binding of Gag, but not of MA, to membranes is cooperative and identifies SPA as a major factor that controls this cooperativity.
Diverse Array of New Viral Sequences Identified in Worldwide Populations of the Asian Citrus Psyllid (Diaphorina citri) Using Viral Metagenomics
Authors: Nouri Shahideh , Salem Nidá , Nigg Jared C. , Falk Bryce W. 
Published: December 2015

ABSTRACT

The Asian citrus psyllid,
Diaphorina citri
, is the natural vector of the causal agent of Huanglongbing (HLB), or citrus greening disease. Together; HLB and
D. citri
represent a major threat to world citrus production. As there is no cure for HLB, insect vector management is considered one strategy to help control the disease, and
D. citri
viruses might be useful. In this study, we used a metagenomic approach to analyze viral sequences associated with the global population of
D. citri
. By sequencing small RNAs and the transcriptome coupled with bioinformatics analysis, we showed that the virus-like sequences of
D. citri
are diverse. We identified novel viral sequences belonging to the picornavirus superfamily, the
Reoviridae
,
Parvoviridae
, and
Bunyaviridae
families, and an unclassified positive-sense single-stranded RNA virus. Moreover, a
Wolbachia
prophage-related sequence was identified. This is the first comprehensive survey to assess the viral community from worldwide populations of an agricultural insect pest. Our results provide valuable information on new putative viruses, some of which may have the potential to be used as biocontrol agents.

IMPORTANCE
Insects have the most species of all animals, and are hosts to, and vectors of, a great variety of known and unknown viruses. Some of these most likely have the potential to be important fundamental and/or practical resources. In this study, we used high-throughput next-generation sequencing (NGS) technology and bioinformatics analysis to identify putative viruses associated with
Diaphorina citri
, the Asian citrus psyllid.
D. citri
is the vector of the bacterium causing Huanglongbing (HLB), currently the most serious threat to citrus worldwide. Here, we report several novel viral sequences associated with
D. citri
.
ICP8 Filament Formation Is Essential for Replication Compartment Formation during Herpes Simplex Virus Infection
Authors: Darwish Anthar S. , Grady Lorry M. , Bai Ping , Weller Sandra K. 
Published: December 2015

ABSTRACT
Herpes simplex virus (HSV) dramatically reorganizes the infected-cell nucleus, leading to the formation of prereplicative sites and replication compartments. This process is driven by the essential viral single-stranded DNA (ssDNA) binding protein ICP8, which can form double-helical filaments in the absence of DNA. In this paper, we show that two conserved motifs, FNF (F1142, N1143, and F1144) and FW (F843 and W844), are essential for ICP8 self-interactions, and we propose that the FNF motif docks into the FW region during filament formation. Mammalian expression plasmids bearing mutations in these motifs (FNF and FW) were unable to complement an ICP8-null mutant for growth and replication compartment formation. Furthermore, FNF and FW mutants were able to inhibit wild-type (WT) virus plaque formation and filament formation, whereas a double mutant (FNF-FW) was not. These results suggest that single mutant proteins are incorporated into nonproductive ICP8 filaments, while the double mutant is unable to interact with WT ICP8 and does not interfere with WT growth. Cells transfected with WT ICP8 and the helicase-primase (H/P) complex exhibited punctate nuclear structures that resemble prereplicative sites; however, the FNF and FW mutants failed to do so. Taken together, these results suggest that the FNF and FW motifs are required for ICP8 self-interactions and that these interactions may be important for the formation of prereplicative sites and replication compartments. We propose that filaments or other higher-order structures of ICP8 may provide a scaffold onto which other proteins can be recruited to form prereplicative sites and replication compartments.

IMPORTANCE
For nuclear viruses such as HSV, efficient DNA replication requires the formation of discrete compartments within the infected-cell nucleus in which replication proteins are concentrated and assembled into the HSV replisome. In this paper, we characterize the role of filament formation by the single-stranded DNA binding protein ICP8 in the formation of prereplicative sites and replication compartments. We propose that ICP8 protein filaments generate a protein scaffold for other cellular and viral proteins, resulting in a structure that concentrates both viral DNA and replication proteins. Replication compartments may be similar to other types of cellular membraneless compartments thought to be formed by phase separations caused by low-affinity, multivalent interactions involving proteins and nucleic acids within cells. ICP8 scaffolds could facilitate the formation of replication compartments by mediating interactions with other components of the replication machinery.
Peripheral T Follicular Helper Cells Are the Major HIV Reservoir within Central Memory CD4 T Cells in Peripheral Blood from Chronically HIV-Infected Individuals on Combination Antiretroviral Therapy
Authors: Pallikkuth Suresh , Sharkey Mark , Babic Dunja Z. , Gupta Sachin , Stone Geoffrey W. , Fischl Margaret A. , Stevenson Mario , Pahwa Savita 
Published: December 2015

ABSTRACT

In this study, we examined the peripheral blood (PB) central memory (T
CM
) CD4
+
T cell subsets designated peripheral T follicular helper cells (pTfh cells) and non-pTfh cells to assess HIV permissiveness and persistence. Purified pTfh and non-pTfh cells from healthy HIV-negative donors were tested for HIV permissiveness using green fluorescent protein (GFP)-expressing HIV-1NL4-3/Ba-L, followed by viral reactivation using beads coated with anti-CD3/anti-CD28 monoclonal antibodies. The role of pTfh cells in HIV persistence was analyzed in 12 chronically HIV-1 infected patients before and 48 weeks after initiation of raltegravir-containing combination antiretroviral therapy (cART). Total cellular HIV-1 DNA and episomes containing two copies of the viral long terminal repeat (2LTR circles) were analyzed in using droplet digital PCR in the purified pTfh and non-pTfh cells. Activation-inducible HIV p24 expression was determined by flow cytometry. Results indicate that pTfh cells, in particular PD1
+
pTfh cells, showed greater permissiveness for HIV infection than non-pTfh cells. At week 48 on cART, HIV DNA levels were unchanged from pre-cART levels, although a significant decrease in 2LTR circles was observed in both cell subsets. Inducible HIV p24 expression was higher in pTfh cells than in non-pTfh cells, with the highest frequencies in the PD1
+
CXCR3
−
pTfh cell subset. Frequencies of HLADR
+
CD38
+
activated CD4 T cells correlated with 2LTR circles in pTfh and non-pTfh cells at both time points and with p24
+
cells at entry. In conclusion, among CD4 T
CM
cells in PB of aviremic patients on cART, pTfh cells, in particular the PD1
+
CXCR3
−
subset, constitute a major HIV reservoir that is sustained by ongoing residual immune activation. The inducible HIV p24 assay is useful for monitoring HIV reservoirs in defined CD4 T cell subsets.

IMPORTANCE
Identification of the type and nature of the cellular compartments of circulating HIV reservoirs is important for targeting of HIV cure strategies. In lymph nodes (LN), a subset of CD4 T cells called T follicular helper (Tfh) cells are preferentially infected by HIV. Central memory (T
CM
) CD4 T cells are the major cellular reservoir for HIV in peripheral blood and contain a subset of CD4 T
CM
cells expressing chemokine receptor CXCR5 similar in function to LN Tfh cells termed peripheral Tfh (pTfh) cells. We found that the circulating pTfh cells are highly susceptible to HIV infection and that in HIV-infected patients, HIV persists in these cells following plasma virus suppression with potent cART. These pTfh cells, which constitute a subset of T
CM
CD4 T cells, can be readily monitored in peripheral blood to assess HIV persistence.
Short biceps femoris fascicles and eccentric knee flexor weakness increase the risk of hamstring injury in elite football (soccer): a prospective cohort study
Authors: Timmins Ryan G , Bourne Matthew N , Shield Anthony J , Williams Morgan D , Lorenzen Christian , Opar David A 
Published: December 2015

Background/aimTo investigate the role of eccentric knee flexor strength, between-limb imbalance and biceps femoris long head (BFlh) fascicle length on the risk of future hamstring strain injury (HSI).MethodsElite soccer players (n=152) from eight different teams participated. Eccentric knee flexor strength during the Nordic hamstring exercise and BFlh fascicle length were assessed at the beginning of preseason. The occurrences of HSIs following this were recorded by the team medical staff. Relative risk (RR) was determined for univariate data, and logistic regression was employed for multivariate data.ResultsTwenty seven new HSIs were reported. Eccentric knee flexor strength below 337 N (RR=4.4; 95% CI 1.1 to 17.5) and possessing BFlh fascicles shorter than 10.56 cm (RR=4.1; 95% CI 1.9 to 8.7) significantly increased the risk of a HSI. Multivariate logistic regression revealed significant effects when combinations of age, history of HSI, eccentric knee flexor strength and BFlh fascicle length were explored. From these analyses the likelihood of a future HSI in older athletes or those with a HSI history was reduced if high levels of eccentric knee flexor strength and longer BFlh fascicles were present.ConclusionsThe presence of short BFlh fascicles and low levels of eccentric knee flexor strength in elite soccer players increases the risk of future HSI. The greater risk of a future HSI in older players or those with a previous HSI is reduced when they have longer BFlh fascicles and high levels of eccentric strength.
Trends in serum creatinine testing in Oxfordshire, UK, 1993–2013: a population-based cohort study
Authors: Oke Jason , Shine Brian , McFadden Emily , Stevens Richard , Lasserson Daniel , Perera Rafael 
Published: December 2015

ObjectivesTo determine how many kidney function tests are done, on whom, how frequently they are performed and how they have changed over time.DesignRetrospective study of all serum creatinine, urine albumin and urine creatinine tests.SettingPrimary and secondary care in Oxfordshire from 1993 to 2013.ParticipantsUnselected population of 1 220 447 people.Main outcome measuresThe total number of creatinine and urinary protein tests ordered from primary and secondary care and the number of tests per year stratified by categories of estimated glomerular filtration rate (eGFR). The frequency of testing in patients having their kidney function monitored.ResultsCreatinine requests from primary care increased steadily from 1997 and exceeded 220 000 requests in 2013. Tests corresponding to normal kidney function (eGFR >60/mL/min/1.73 m2) constituted 59% of all kidney function tests in 1993 and accounted for 83% of all tests in 2013. Test corresponding to chronic kidney disease (CKD) stages 3–5 declined after 2007. Reduced kidney function, albuminuria, male gender, diabetes and age were independently associated with more frequent monitoring. For a female patient between 61 and 80 years and with stage 3a CKD, the average number of serum creatinine tests (95% CI) was 3.23/year (3.19 to 3.26) and for a similar woman with diabetes, the average number of tests was 5.50 (5.44 to 5.56) tests per year.ConclusionsThere has been a large increase in the number of kidney function tests over the past two decades. However, we found little evidence that this increase is detecting more CKD. Tests are becoming more frequent in people with and without evidence of renal impairment. Future work using a richer data source could help unravel the underlying reasons for the increased testing and determine how much is necessary and useful.
Prolonged seizure‐like phenomena in an English springer spaniel after total intravenous anaesthesia with propofol
Authors: Monticelli Paolo , Seymour Christopher John 
Published: December 2015

Propofol is an anaesthetic agent commonly used for both induction and maintenance of general anaesthesia and has been associated with seizure‐like phenomena (SLP) in both veterinary and human medicine. This case report describes an English springer spaniel with no previous history of neurological symptoms, which exhibited SLP after propofol anaesthesia for bronchoscopy. The SLP were refractory to treatment for 13 hours, but steadily improved after administration of phenobarbital and levetiracetam. To the authors’ knowledge, this is the first report of their use to manage SLP.
Research on sintering behavior and microwave dielectric property of (Mg0.95Ca0.05)TiO3 ceramics for cross coupling filter
Authors: Luo Chunya , Ma Zhichao , Hu Laisheng , Hu Mingzhe , Huang Xiaomin 
Published: December 2015

The microwave dielectric properties of [Formula: see text]–[Formula: see text] (abbreviated as 95MCT hereafter) ceramics have been studied for application in dielectric cross coupling filters. [Formula: see text] and [Formula: see text] were selected as liquid sintering aids to lower the sintering temperature and enhance the Qf value of 95MCT and simultaneously we varied the mole ratio of [Formula: see text] to tune the microwave dielectric properties of 95MCT. When the [Formula: see text] mole ratio was 1.5 and the co-doping content was 0.25 wt.%, the optimal sintering temperature of 95MCT ceramic could be lowered from [Formula: see text] to [Formula: see text] and the Qf value could be improved by about 7.7%. The optimal microwave dielectric properties obtained under this condition were Qf = 72730 GHz (6.8 GHz), [Formula: see text] and [Formula: see text], which demonstrated great potential usage in ceramic industry. High values of Qf ceramic were used to design the dielectric cross coupling filter. The dielectric filter measured at 2.35 GHz exhibited a 6.7% bandwidth (insert loss [Formula: see text] dB) of center frequency.
The vertebrate-bearing Late Triassic Fleming Fjord Formation of central East Greenland revisited: stratigraphy, palaeoclimate and new palaeontological data
Authors: Clemmensen Lars B. , Milàn Jesper , Adolfssen Jan Schulz , Estrup Eliza Jarl , Frobøse Nicolai , Klein Nicole , Mateus Octávio , Wings Oliver 
Published: December 2015

Abstract
In Late Triassic (Norian–Rhaetian) times, the Jameson Land Basin lay at 40° N on the northern part of the supercontinent Pangaea. This position placed the basin in a transition zone between the relatively dry interior of the supercontinent and its more humid periphery. Sedimentation in the Jameson Land Basin took place in a lake–mudflat system and was controlled by orbitally forced variations in precipitation. Vertebrate fossils have consistently been found in these lake deposits (Fleming Fjord Formation), and include fishes, dinosaurs, amphibians, turtles, aetosaurs and pterosaurs. Furthermore, the fauna includes mammaliaform teeth and skeletal material. New vertebrate fossils were found during a joint vertebrate palaeontological and sedimentological expedition to Jameson Land in 2012. These new finds include phytosaurs, a second stem testudinatan specimen and new material of sauropodomorph dinosaurs, including osteologically immature individuals. Phytosaurs are a group of predators common in the Late Triassic, but previously unreported from Greenland. The finding includes well-preserved partial skeletons that show the occurrence of four individuals of three size classes. The new finds support a late Norian–early Rhaetian age for the Fleming Fjord Formation, and add new information on the palaeogeographical and palaeolatitudinal distribution of Late Triassic faunal provinces.
Novel Morphologic and Genetic Analysis of Cancer Cells in a 3D Microenvironment Identifies STAT3 as a Regulator of Tumor Permeability Barrier Function
Authors: Park Min Chul , Jeong Hyobin , Son Sung Hwa , Kim YounHa , Han Daeyoung , Goughnour Peter C. , Kang Taehee , Kwon Nam Hoon , Moon Hyo Eun , Paek Sun Ha , Hwang Daehee , Seol Ho Jun , Nam Do-Hyun , Kim Sunghoon 
Published: December 2015

Abstract
Tumor permeability is a critical determinant of drug delivery and sensitivity, but systematic methods to identify factors that perform permeability barrier functions in the tumor microenvironment are not yet available. Multicellular tumor spheroids have become tractable in vitro models to study the impact of a three-dimensional (3D) environment on cellular behavior. In this study, we characterized the spheroid-forming potential of cancer cells and correlated the resulting spheroid morphologies with genetic information to identify conserved cellular processes associated with spheroid structure. Spheroids generated from 100 different cancer cell lines were classified into four distinct groups based on morphology. In particular, round and compact spheroids exhibited highly hypoxic inner cores and permeability barriers against anticancer drugs. Through systematic and correlative analysis, we reveal JAK–STAT signaling as one of the signature pathways activated in round spheroids. Accordingly, STAT3 inhibition in spheroids generated from the established cancer cells and primary glioblastoma patient–derived cells altered the rounded morphology and increased drug sensitivity. Furthermore, combined administration of the STAT3 inhibitor and 5-fluorouracil to a mouse xenograft model markedly reduced tumor growth compared with monotherapy. Collectively, our findings demonstrate the ability to integrate 3D culture and genetic profiling to determine the factors underlying the integrity of the permeability barrier in the tumor microenvironment, and may help to identify and exploit novel mechanisms of drug resistance. Cancer Res; 76(5); 1044–54. ©2015 AACR.