TitleAbstractYear(sorted descending)
a survey of mosquito-borne and insect-specific viruses in hospitals and livestock markets in western kenya.aedes aegypti and culex pipiens complex mosquitoes are prolific vectors of arboviruses that are a global threat to human and animal health. increased globalization and ease of travel have facilitated the worldwide dissemination of these mosquitoes and the viruses they transmit. to assess disease risk, we determined the frequency of arboviruses in western kenyan counties bordering an area of high arboviral activity. in addition to pathogenic viruses, insect-specific flaviviruses (isfs), some of w ...202134048473
targeting lentiviral vectors to primordial germ cells (pgcs): an efficient strategy for generating transgenic chickens.recent advances in avian transgenic studies highlight the possibility of utilizing lentiviral vectors as tools to generate transgenic chickens. however, low rates of gonadal chimerism and germ line transmission efficiency still limit the broad usage of this method in creating transgenic chickens. in this study, we implemented a simple strategy using modified lentiviral vectors targeted to chicken primordial germ cells (pgcs) to generate transgenic chickens. the lentiviral vectors were pseudotype ...202032274905
arboviruses and blood meal sources in zoophilic mosquitoes at human-wildlife interfaces in kenya.background: zoophilic mosquitoes play an important role in the transmission of arboviruses of medical importance at human-wildlife interfaces, yet arbovirus surveillance efforts have been focused mostly on anthropophilic mosquitoes. understanding the diversity of zoophilic mosquitoes and their associated feeding patterns and arboviruses can inform better vector control strategies. materials and methods: we morphologically identified mosquitoes collected from two game reserves in kenya, the maasa ...202032155389
infection of western gray kangaroos (macropus fuliginosus) with australian arboviruses associated with human infection.more than 75 arboviruses (arthropod-borne viruses) have been identified in australia. while alfuy virus (alfv), barmah forest virus (bfv), edge hill virus (ehv), kokobera virus (kokv), murray valley encephalitis virus (mvev), sindbis virus (sinv), ross river virus (rrv), stratford virus (strv), and west nile virus strain kunjin (kunv) have been associated with human infection, there remains a paucity of data regarding their respective transmission cycles and any potential nonhuman vertebrate hos ...202031556842
common vole (microtus arvalis) and bank vole (myodes glareolus) derived permanent cell lines differ in their susceptibility and replication kinetics of animal and zoonotic viruses.pathogenesis and reservoir host adaptation of animal and zoonotic viruses are poorly understood due to missing adequate cell culture and animal models. the bank vole (myodes glareolus) and common vole (microtus arvalis) serve as hosts for a variety of zoonotic pathogens. for a better understanding of virus association to a putative animal host, we generated two novel cell lines from bank voles of different evolutionary lineages and two common vole cell lines and assayed their susceptibility, rep ...201931513859
sindbis virus- a wild bird associated zoonotic arbovirus circulates in germany.sindbis virus (sinv) is an arbovirus causing clinical symptoms such as arthritis, rash and fever following human infections in fennoscandia. its transmission cycle involves mosquito species as vectors as well as wild birds that act as natural reservoir hosts. in germany, sinv was first time observed in 2009 in different mosquito species in the upper rhine valley and one year later in a hooded crow in berlin. recently, sinv was also detected repeatedly at various locations in germany in the conte ...201931767092
activation of rnase l in egyptian rousette bat-derived roni/7 cells is dependent primarily on oas3 and independent of mavs signaling.bats are reservoirs for many rna viruses that are highly pathogenic in humans yet relatively apathogenic in the natural host. it has been suggested that differences in innate immunity are responsible. the antiviral oas-rnase l pathway is well characterized in humans, but there is little known about its activation and antiviral activity in bats. during infection, oass, upon sensing double-stranded rna (dsrna), produce 2'-5' oligoadenylates (2-5a), leading to activation of rnase l which degrades v ...201931719180
linkages between chiropteran diversity and ecosystem services for sustainable fragmented forest conservation.this data article informs about chiropteran diversity, new records, ecosystem services and possible pathogen carriers in fragmented forests (sub-divided by utility corridors, man-made structures, untouched and secondary plantations) within districts setiu (setiu research station), hulu terengganu (saok and lasir waterfalls) and besut (gunung tebu forest reserve) of state terengganu, peninsular malaysia. these bats were captured using harp traps and mist nets that were set 10 m apart across flywa ...201830533456
host-feeding patterns of culex mosquitoes in iran.different mosquito-borne pathogens are circulating in iran including sindbis virus, west nile virus, filarioid worms and malaria parasites. however, the local transmission cycles of these pathogenic agents are poorly understood, especially because ecological data on vector species are scarce and there is limited knowledge about the host range; this understanding could help to direct species-specific vector control measurements or to prioritize research.201830587194
arboviruses isolated from the barkedji mosquito-based surveillance system, 2012-2013.a mosquito-based arbovirus surveillance system was set up at barkedji, senegal after the first outbreak of rift valley fever in west africa in 1988. this system was recently updated using more sampling methods and collecting in greater number of ponds and villages sites.201830541472
imaging of viral neuroinvasion in the zebrafish reveals that sindbis and chikungunya viruses favour different entry routes.alphaviruses, such as chikungunya virus (chikv) and sindbis virus (sinv), are vector-borne pathogens that cause acute illnesses in humans and are sometimes associated with neuropathies, especially in infants and elderly patients. little is known about their mechanism of entry into the central nervous system (cns), even for sinv, which has been used extensively as a model for viral encephalopathies. we previously established a chikv infection model in the optically transparent zebrafish larva; he ...201728483796
identification of a newly isolated getah virus in the china-laos border, this study, we isolated a virus strain (yn12031) from specimens of armigeres subalbatus collected in the china-laos border. bhk-21 cells infected with yn12031 exhibited an evident cytopathic effect (cpe) 32 h post-infection. the virus particles were spherical, 70 nm in diameter, and enveloped; they also featured surface fibers. molecular genetic analysis revealed that yn12031 was closely related to alpha viruses such as chikungunya virus and sindbis virus, and located in the same clade as mm2 ...201728427491
biological safety of a highly purified 10% liquid intravenous immunoglobulin preparation from human plasma.a highly purified 10% liquid intravenous immunoglobulin, iqymune(®), has been developed using an innovative manufacturing process including an affinity chromatography step for the removal of anti-a and anti-b hemagglutinins.201728508264
partially uncleaved alphavirus replicase forms spherule structures in the presence and absence of rna template.alphaviruses are positive-strand rna viruses expressing their replicase as a polyprotein, p1234, which is cleaved to four final products, nonstructural proteins nsp1 to nsp4. the replicase proteins together with viral rna and host factors form membrane invaginations termed spherules, which act as the replication complexes producing progeny rnas. we have previously shown that the wild-type alphavirus replicase requires a functional rna template and active polymerase to generate spherule structure ...201728701392
application of alphaviral vectors for immunomodulation in cancer therapy.the lack of specific and efficient cancer therapies has influenced the development of novel approaches, such as immunotherapy, which from its original application of immunogenic protein delivery has developed into the use of more sophisticated recombinant gene delivery methods to achieve better safety and efficacy profiles. this approach involves viral and non-viral delivery systems.201728641531
oncolytic alphaviruses in cancer immunotherapy.oncolytic viruses show specific targeting and killing of tumor cells and therefore provide attractive assets for cancer immunotherapy. in parallel to oncolytic viral vectors based on adenoviruses and herpes simplex viruses, oncolytic rna viruses and particularly alphaviruses have been evaluated as delivery vehicles. immunization studies in experimental rodent models for various cancers including glioblastoma, hematologic, hepatocellular, colon, cervix, and lung cancer as well as melanoma have be ...201728417936
h-ras exerts opposing effects on type i interferon responses depending on its activation status.using shrna high-throughput screening, we identified h-ras as a regulator of antiviral activity, whose depletion could enhance sindbis virus replication. further analyses indicated that depletion of h-ras results in a robust increase in vesicular stomatitis virus infection and a decrease in sendai virus (sev)-induced retinoic acid-inducible gene-i-like receptor (rlr) signaling. interestingly, however, ectopic expression of wild-type h-ras results in a biphasic mode of rlr signaling regulation: w ...201728848563
dorsal and ventral parts of thalamic nucleus submedius project to different areas of rat orbitofrontal cortex: a single neuron-tracing study using virus vectors.the rodent orbitofrontal cortex is involved in a variety of cognitive and behavioral functions that require thalamic input to be successfully expressed. although the thalamic nucleus submedius (sm) is a major source of afferents to the orbitofrontal cortex, thalamocortical projection from the sm has not been fully elucidated. in the present study, we first divided the rat sm into dorsal and ventral parts according to the distribution of vesicular glutamate transporter 2-immunoreactive varicositi ...201728833113
identification of interactions between sindbis virus capsid protein and cytoplasmic vrna as novel virulence determinants.alphaviruses are arthropod-borne viruses that represent a significant threat to public health at a global level. while the formation of alphaviral nucleocapsid cores, consisting of cargo nucleic acid and the viral capsid protein, is an essential molecular process of infection, the precise interactions between the two partners are ill-defined. a clip-seq approach was used to screen for candidate sites of interaction between the viral capsid protein and genomic rna of sindbis virus (sinv), a model ...201728662211
wolbachia elevates host methyltransferase expression to block an rna virus early during infection.wolbachia pipientis is an intracellular endosymbiont known to confer host resistance against rna viruses in insects. however, the causal mechanism underlying this antiviral defense remains poorly understood. to this end, we have established a robust arthropod model system to study the tripartite interaction involving sindbis virus and wolbachia strain wmel within its native host, drosophila melanogaster. by leveraging the power of drosophila genetics and a parallel, highly tractable d. melanogas ...201728617844
development and characterization of sindbis virus with encoded fluorescent rna aptamer spinach2 for imaging of replication and immune-mediated changes in intracellular viral rna.viral rna studies often rely on in situ hybridization and reverse transcriptase-pcr to provide snapshots of rna dynamics in infected cells. to facilitate analysis of cellular rnas, aptamers spinach and spinach2 that bind and activate the conditional fluorophore 3, 5-difluoro-4-hydroxybenzylidene imidazolinon have been developed. to determine the feasibility of applying this technology to viral rna, we have used cdna clones of the te strain of sindbis virus (sinv) to construct multiple viruses co ...201728555544
glutamine antagonist-mediated immune suppression decreases pathology but delays virus clearance in mice during nonfatal alphavirus encephalomyelitis.infection of weanling c57bl/6 mice with the te strain of sindbis virus (sinv) causes nonfatal encephalomyelitis associated with hippocampal-based memory impairment that is partially prevented by treatment with 6-diazo-5-oxo-l-norleucine (don), a glutamine antagonist (potter et al., j neurovirol 21:159, 2015). to determine the mechanism(s) of protection, lymph node and central nervous system (cns) tissues from sinv-infected mice treated daily for 1 week with low (0.3mg/kg) or high (0.6mg/kg) dose ...201728531865
operational mosquito and vector-borne diseases surveillance at incirlik air base, turkey.arboviruses on incirlik air base, turkey, pose a threat to military personnel and civilians, but might also be relevant for understanding the threats in neighboring conflict zones such as syria. we reviewed 6 years of mosquito and arbovirus surveillance at incirlik air base. over 6,000 mosquitoes were identified as aedes caspius, anopheles claviger, culex mimeticus, cx. perexiguus, cx. pipiens, cx. sinaiticus, and culiseta longiareolata. almost all of the mosquitoes (more than 90%) were cx. pere ...201728511277
individual mediodorsal thalamic neurons project to multiple areas of the rat prefrontal cortex: a single neuron-tracing study using virus vectors.the prefrontal cortex has an important role in a variety of cognitive and executive processes, and is generally defined by its reciprocal connections with the mediodorsal thalamic nucleus (md). the rat md is mainly subdivided into three segments, the medial (mdm), central (mdc), and lateral (mdl) divisions, on the basis of the cytoarchitecture and chemoarchitecture. the md segments are known to topographically project to multiple prefrontal areas at the population level: the mdm mainly to the pr ...201727275581
alphavirus-based vaccines.alphavirus-based vectors have been engineered from semliki forest virus, sindbis virus, and venezuelan equine encephalitis virus and applied for vaccine development. immunization in preclinical animal models has been conducted with naked rna replicons, recombinant viral particles and layered dna-rna vectors. most commonly, the targets for the immunization have been viral surface proteins and tumor antigens, which have elicited strong immune responses and even provided protection against challeng ...201728374252
discovery of a novel antiviral agent targeting the nonstructural protein 4 (nsp4) of chikungunya virus.chikungunya fever (chikf) is caused by chikungunya virus (chikv) infection which is a re-emerging mosquito-borne zoonosis. at present, there are no approved therapeutics for chikf. herein, we have investigated candidate compounds which can inhibit chikv infection. screening of chemical compound libraries were performed and one candidate, a benzimidazole-related compound designated compound-a was found to inhibit infection by several chikv strains and a sindbis virus strain at nanomolar concentra ...201728236746
selective transduction of astrocytic and neuronal cns subpopulations by lentiviral vectors pseudotyped with chikungunya virus envelope.lentiviral vectors are gene delivery vehicles that integrate into the host genome of dividing and non-dividing mammalian cells facilitating long-term transgene expression. lentiviral vector versatility is greatly increased by incorporating heterologous viral envelope proteins onto the vector particles instead of the native envelope, conferring on these pseudotyped vectors a modified tropism and host range specificity. we investigated the pseudotyping efficiency of hiv-1 based lentiviral vectors ...201728152379
obatoclax inhibits alphavirus membrane fusion by neutralizing the acidic environment of endocytic new pathogenic viruses continue to emerge, it is paramount to have intervention strategies that target a common denominator in these pathogens. the fusion of viral and cellular membranes during viral entry is one such process that is used by many pathogenic viruses, including chikungunya virus, west nile virus, and influenza virus. obatoclax, a small-molecule antagonist of the bcl-2 family of proteins, was previously determined to have activity against influenza a virus and also sindbis virus ...201727993855
ablation of programmed -1 ribosomal frameshifting in venezuelan equine encephalitis virus results in attenuated neuropathogenicity.the alphaviruses venezuelan equine encephalitis virus (veev), eastern equine encephalitis virus (eeev), and western equine encephalitis virus (weev) are arthropod-borne positive-strand rna viruses that are capable of causing acute and fatal encephalitis in many mammals, including humans. veev was weaponized during the cold war and is recognized as a select agent. currently, there are no fda-approved vaccines or therapeutics for these viruses. the spread of veev and other members of this family d ...201727852852
trim25 is required for the antiviral activity of zinc-finger antiviral protein (zap).zinc-finger antiviral protein (zap) is a host factor that specifically inhibits the replication of certain viruses by binding to viral mrnas, and repressing the translation and/or promoting the degradation of target mrna. in addition, zap regulates the expression of certain cellular genes. here, we report that tripartite motif-containing protein 25 (trim25), a ubiquitin e3 ligase, is required for the antiviral activity of zap. downregulation of endogenous trim25 abolished zap's antiviral activit ...201728202764
inactivation of viruses by pasteurization at 60 °c for 10 h with and without 40% glucose as stabilizer during a new manufacturing process of α2-macroglobulin from cohn fraction iv.pasteurization is regularly used to inactivate viruses for the safety of plasma derivatives. influence of pasteurization at 60 °c for 10 h on α2-macroglobulin activity and virus inactivation were studied. with 40% sugar as stabilizers more than 70% α2-macroglobulin activity was reserved after pasteurization compared with 20% in control. glucose presented a better activity protection effect than sucrose and maltose. by pasteurization without stabilizer the virus titers of pseudorabies virus, sind ...201728215695
an icosahedral virus as a fluorescent calibration standard: a method for counting protein molecules in cells by fluorescence microscopy.the ability to replace genes coding for cellular proteins with dna that codes for fluorescent protein-tagged versions opens the way to counting the number of molecules of each protein component of macromolecular assemblies in vivo by measuring fluorescence microscopically. converting fluorescence to absolute numbers of molecules requires a fluorescent standard whose molecular composition is known precisely. in this report, the construction, properties and mode of using a set of fluorescence cali ...201728328099
translation of sindbis subgenomic mrna is independent of eif2, eif2a and eif2d.translation of sindbis virus subgenomic mrna (sgmrna) can occur after inactivation of eif2 by phosphorylation in mammalian cells. several studies have suggested that eif2 can be replaced by eif2a or eif2d. hap1 human cell lines knocked-out for eif2a, eif2d or both by crispr/cas9 genome engineering were compared with wild-type (wt) cells to test the potential role of eif2a and eif2d in translation. sindbis virus infection was comparable between the four cell lines. moreover, synthesis of viral pr ...201728240315
microbiota activates imd pathway and limits sindbis infection in aedes aegypti.aedes aegypti is the main vector of important arboviruses such as dengue, zika and chikungunya. during infections mosquitoes can activate the immune pathways toll, imd and jak/stat to limit pathogen replication.201728231846
mitochondrial damage elicits a tcdd-inducible poly(adp-ribose) polymerase-mediated antiviral response.the innate immune system senses rna viruses by pattern recognition receptors (prrs) and protects the host from virus infection. prrs mediate the production of immune modulatory factors and direct the elimination of rna viruses. here, we show a unique prr that mediates antiviral response. tetrachlorodibenzo-p-dioxin (tcdd)-inducible poly(adp ribose) polymerase (tiparp), a cysteine3 histidine (ccch)-type zinc finger-containing protein, binds to sindbis virus (sinv) rna via its zinc finger domain a ...201728213497
spatial and temporal analysis of alphavirus replication and assembly in mammalian and mosquito cells.sindbis virus (sinv [genus alphavirus, family togaviridae]) is an enveloped, mosquito-borne virus. alphaviruses cause cytolytic infections in mammalian cells while establishing noncytopathic, persistent infections in mosquito cells. mosquito vector adaptation of alphaviruses is a major factor in the transmission of epidemic strains of alphaviruses. though extensive studies have been performed on infected mammalian cells, the morphological and structural elements of alphavirus replication and ass ...201728196962
dynamics of molecular evolution in rna virus populations depend on sudden versus gradual environmental change.understanding the dynamics of molecular adaptation is a fundamental goal of evolutionary biology. while adaptation to constant environments has been well characterized, the effects of environmental complexity remain seldom studied. one simple but understudied factor is the rate of environmental change. here we used experimental evolution with rna viruses to investigate whether evolutionary dynamics varied based on the rate of environmental turnover. we used whole-genome next-generation sequencin ...201728121018
trim25 enhances the antiviral action of zinc-finger antiviral protein (zap).the host factor and interferon (ifn)-stimulated gene (isg) product, zinc-finger antiviral protein (zap), inhibits a number of diverse viruses by usurping and intersecting with multiple cellular pathways. to elucidate its antiviral mechanism, we perform a loss-of-function genome-wide rnai screen to identify cellular cofactors required for zap antiviral activity against the prototype alphavirus, sindbis virus (sinv). in order to exclude off-target effects, we carry out stringent confirmatory assay ...201728060952
neurotropic alphaviruses can propagate without capsid. 201728129638
palmitoylation of sindbis virus tf protein regulates its plasma membrane localization and subsequent incorporation into virions.palmitoylation is a reversible, posttranslational modification that helps target proteins to cellular membranes. the alphavirus small membrane proteins 6k and tf have been reported to be palmitoylated and to positively regulate budding. 6k and tf are isoforms that are identical in their n termini but unique in their c termini due to a -1 ribosomal frameshift during translation. in this study, we used cysteine (cys) mutants to test differential palmitoylation of the sindbis virus 6k and tf protei ...201727852864
sindbis virus-pseudotyped lentiviral vectors carrying vegfr2-specific nanobody for potential transductional targeting of tumor vasculature.introduction of selectivity/specificity into viral-based gene delivery systems, such as lentiviral vectors (lvs), is crucial in their systemic administration for cancer gene therapy. the pivotal role of tumor-associated endothelial cells (taecs) in tumor angiogenesis and overexpression of vascular endothelial growth factor receptor-2 (vegfr2 or kdr) in taecs makes them a potent target in cancer treatment. herein, we report the development of vegfr2-targeted lvs pseudotyped with chimeric sindbis ...201627647452
sindbis virus can exploit a host antiviral protein to evade immune surveillance.viral infection induces production of type i interferons (ifns), which stimulate the expression of a variety of antiviral factors to inhibit viral replication. to establish effective infection, viruses need to develop strategies to evade the immune responses. a neurovirulent sindbis virus strain with neuroinvasive properties (svni) causes lethal encephalitis in mice, and its replication in cultured cells is inhibited by the zinc finger antiviral protein (zap), a host factor that specifically inh ...201627581990
wide recognition of culex pipiens and lack of detection of culex torrentium through biomolecular differentiation of mosquitoes in the emilia-romagna region, northern italy.the culex pipiens complex includes species with reported differences in vector competence for arthropod-borne viruses, many of which are of significant importance to human health such as the west nile virus and the sindbis virus. this group of mosquitoes is difficult to distinguish morphologically; particularly as adult females. in europe, the two species of the complex, culex pipiens linnaeus 1758 and culex torrentium martini 1925, are often found sympatrically. with the aim to characterize the ...201627477051
interferon gamma modulation of disease manifestation and the local antibody response to alphavirus encephalomyelitis.infection of mice with sindbis virus (sinv) produces encephalomyelitis and provides a model for examination of the central nervous system (cns) immune response to alphavirus infection. clearance of infectious virus is accomplished through a cooperative effort between sinv-specific antibody and ifn-γ, but the regulatory interactions are poorly understood. to determine the effects of ifn-γ on clinical disease and the antiviral immune response, c57bl/6 mice lacking ifn-γ (ifng-/-) or ifn-γ receptor ...201627667782
protective effects of glutamine antagonist 6-diazo-5-oxo-l-norleucine in mice with alphavirus encephalomyelitis.inflammation is a necessary part of the response to infection but can also cause neuronal injury in both infectious and autoimmune diseases of the central nervous system (cns). a neurovirulent strain of sindbis virus (nsv) causes fatal paralysis in adult c57bl/6 mice during clearance of infectious virus from the cns, and the virus-specific immune response is implicated as a mediator of neuronal damage. previous studies have shown that survival is improved in t-cell-deficient mice and in mice wit ...201627489275
full inactivation of alphaviruses in single particle and crystallized forms.inherent in the study of viruses is the risk of pathogenic exposure, which necessitates appropriate levels of biosafety containment. unfortunately, this also limits the availability of useful research instruments that are located at facilities not equipped to handle infectious pathogens. abrogation of viral infectivity can be accomplished without severely disrupting the physical structure of the virus particle. virus samples that are verifiably intact but not infectious may be enabled for study ...201627465218
capsid-deficient alphaviruses generate propagative infectious microvesicles at the plasma membrane.alphavirus budding is driven by interactions between nucleocapsids assembled in the cytoplasm and envelope proteins present at the plasma membrane. so far, the expression of capsid and envelope proteins in infected cells has been considered an absolute requirement for alphavirus budding and propagation. in the present study, we show that semliki forest virus and sindbis virus lacking the capsid gene can propagate in mammalian and insect cells. this propagation is mediated by the release of infec ...201627117550
interferon regulatory factor 2 protects mice from lethal viral neuroinvasion.the host responds to virus infection by activating type i interferon (ifn) signaling leading to expression of ifn-stimulated genes (isgs). dysregulation of the ifn response results in inflammatory diseases and chronic infections. in this study, we demonstrate that ifn regulatory factor 2 (irf2), an isg and a negative regulator of ifn signaling, influences alphavirus neuroinvasion and pathogenesis. a sindbis virus strain that in wild-type (wt) mice only causes disease when injected into the brain ...201627899441
identification of aadnr1, a novel gene related to innate immunity and apoptosis in aedes albopictus.innate immunity and apoptosis play critical roles in defending pathogens in insects. in drosophila, dnr1 was reported as a negative regulator of apoptosis and immune deficiency (imd) pathway which belongs to innate immunity. aedes albopictus is an important kind of arbovirus vector and becoming a significant threat to public health due to its rapid global expansion. here we identified an ortholog of dnr1 from a. albopictus, named as aadnr1. aadnr1 encoded a putative protein containing an n-termi ...201627045774
single-site glycoprotein mutants inhibit a late event in sindbis virus assembly.a panel of sindbis virus mutants that were suspected to have deficiencies in one or more aspects of their replication cycles was examined in baby hamster kidney (bhk) cells. these included an amino acid deletion (δh230) and substitution (h230a) in the sindbis glycoprotein e1_h230 and similar mutants in e2_g209 (g209a, g209d, and δg209). neither h230 mutation produced a measurable titer, but repeated passaging of the h230a mutant in bhk cells produced a second-site compensatory mutant (v231i) tha ...201627412592
alphavirus protease inhibitors from natural sources: a homology modeling and molecular docking investigation.alphaviruses such as chikungunya virus (chikv), o'nyong-nyong virus (onnv), ross river virus (rrv), eastern equine encephalitis virus (eeev), venezuelan equine encephalitis virus (veev), and western equine encephalitis virus (weev), are mosquito-transmitted viruses that can cause fevers, rash, and rheumatic diseases (chikv, onnv, rrv) or potentially fatal encephalitis (eeev, veev, weev) in humans. these diseases are considered neglected tropical diseases for which there are no current antiviral ...201627387412
antiviral activities of niclosamide and nitazoxanide against chikungunya virus entry and transmission.chikungunya disease results from an infection with the arbovirus, chikungunya virus (chikv). symptoms of chikv include fever and persistent, severe arthritis. in recent years, several antiviral drugs have been evaluated in clinical trials; however, no registered antivirals have been approved for clinical therapy. in this study, we established a high-throughput screening (hts) system based on chikv 26s mediated insect cell fusion inhibition assay. our screening system was able to search potential ...201627742486
inhibition of chikungunya virus by picolinate that targets viral capsid protein.the protein-protein interactions (ppis) of the transmembrane glycoprotein e2 with the hydrophobic pocket on the surface of capsid protein (cp) plays a critical role in alphavirus life cycle. dioxane based derivatives targeting ppis have been reported to possess antiviral activity against sindbis virus (sinv), the prototype alphavirus. in this study, the binding of picolinic acid (pca) to the conserved hydrophobic pocket of capsid protein was analyzed by molecular docking, isothermal titration ca ...201627614702
mapping of human b-cell epitopes of sindbis virus.mosquito-transmitted sindbis virus (sinv) causes fever, skin lesions and musculoskeletal symptoms if transmitted to man. sinv is the prototype virus of genus alphavirus, which includes other arthritogenic viruses such as chikungunya virus (chikv) and ross river virus (rrv) that cause large epidemics with a considerable public health burden. until now the human b-cell epitopes have been studied for chikv and rrv, but not for sinv. to identify the b-cell epitopes in sinv-infection, we synthetised ...201627339177
host translation shutoff mediated by non-structural protein 2 is a critical factor in the antiviral state resistance of venezuelan equine encephalitis virus.most previous studies of interferon-alpha/beta (ifn-α/β) response antagonism by alphaviruses have focused upon interruption of ifn-α/β induction and/or receptor signaling cascades. infection of mice with venezuelan equine encephalitis alphavirus (veev) or sindbis virus (sinv) induces serum ifn-α/β, that elicits a systemic antiviral state in uninfected cells successfully controlling sinv but not veev replication. furthermore, veev replication is more resistant than that of sinv to a pre-existing ...201627318152
alphavirus-based vaccines.alphavirus vectors based on semliki forest virus, sindbis virus, and venezuelan equine encephalitis virus have been widely applied for vaccine development. naked rna replicons, recombinant viral particles, and layered dna vectors have been subjected to immunization in preclinical animal models with antigens for viral targets and tumor antigens. moreover, a limited number of clinical trials have been conducted in humans. vaccination with alphavirus vectors has demonstrated efficient immune respon ...201627076308
molecular determinants of alphavirus neuropathogenesis in mice.alphaviruses are enveloped viruses with a positive-stranded rna genome, of the family togaviridae. in mammals and birds they are mosquito-transmitted and are of veterinary and medical importance. they cause primarily two types of disease: encephalitis and polyarthritis. here we review attempts to understand the molecular basis of encephalitis and virulence for the central nervous system (cns) in mouse models. sindbis virus (sinv) was the first virus to be studied in this way. other viruses analy ...201627028153
the presence and seroprevalence of arthropod-borne viruses in nasiriyah governorate, southern iraq: a cross-sectional study.the knowledge on the presence and seroprevalence of arboviruses in iraq is fragmental. to assess the exposure of the population to arbovirus infections in southern iraq, we conducted a serological screening of the most common arbovirus groups using immunofluorescence, hemagglutination inhibition and neutralization tests. serum samples of 399 adult volunteers were collected in nasiriyah, iraq. antibodies were detected against west nile virus (wnv) (11.6%), sandfly-borne sicilian virus serocomplex ...201626880770
discovery of berberine, abamectin and ivermectin as antivirals against chikungunya and other alphaviruses.chikungunya virus (chikv) is an arthritogenic arbovirus of the alphavirus genus, which has infected millions of people after its re-emergence in the last decade. in this study, a bhk cell line containing a stable chikv replicon with a luciferase reporter was used in a high-throughput platform to screen approximately 3000 compounds. following initial validation, 25 compounds were chosen as primary hits for secondary validation with wild type and reporter chikv infection, which identified three pr ...201626752081
an mrna-specific trnai carrier eif2a plays a pivotal role in cell proliferation under stress conditions: stress-resistant translation of c-src mrna is mediated by eif2a.c-src, a non-receptor protein tyrosine kinase, activates nf-κb and stat3, which in turn triggers the transcription of anti-apoptosis- and cell cycle-related genes. c-src protein regulates cell proliferation, cell motility and programmed cell death. and the elevated level of activated c-src protein is related with solid tumor generation. translation of c-src mrna is directed by an ires element which mediates persistent translation under stress conditions when translation of most mrnas is inhibite ...201627899592
use of aptamers as diagnostics tools and antiviral agents for human viruses.appropriate diagnosis is the key factor for treatment of viral diseases. time is the most important factor in rapidly developing and epidemiologically dangerous diseases, such as influenza, ebola and sars. chronic viral diseases such as hiv-1 or hcv are asymptomatic or oligosymptomatic and the therapeutic success mainly depends on early detection of the infective agent. over the last years, aptamer technology has been used in a wide range of diagnostic and therapeutic applications and, concretel ...201627999271
the conserved coronavirus macrodomain promotes virulence and suppresses the innate immune response during severe acute respiratory syndrome coronavirus infection.adp-ribosylation is a common posttranslational modification that may have antiviral properties and impact innate immunity. to regulate this activity, macrodomain proteins enzymatically remove covalently attached adp-ribose from protein targets. all members of the coronavirinae, a subfamily of positive-sense rna viruses, contain a highly conserved macrodomain within nonstructural protein 3 (nsp3). however, its function or targets during infection remain unknown. we identified several macrodomain ...201627965448
first report on human-biting culex pipiens in sweden.culex mosquitoes are vectors of several bird-hosted arboviruses that cause outbreaks in europe, such as sindbis virus and west nile virus. recently, the human-biting form of culex pipiens, cx. pipiens biotype molestus, was found causing big nuisance in a housing cooperative in gothenburg in southern sweden, confirmed by molecular identification. this is the first report of human-biting culex in scandinavia, signalling increased risk of arbovirus infection in northern europe.201627927233
molecular evolution of type 2 porcine reproductive and respiratory syndrome viruses circulating in vietnam from 2007 to 2015.porcine respiratory and reproductive syndrome (prrs) virus is one of the most economically significant pathogens in the vietnamese swine industry. orf5, which participates in many functional processes, including virion assembly, entry of the virus into the host cell, and viral adaptation to the host immune response, has been widely used in molecular evolution and phylogeny studies. knowing of molecular evolution of prrsv fields strains might contribute to prrs control in vietnam.201627855667
yellow fever virus capsid protein is a potent suppressor of rna silencing that binds double-stranded rna.mosquito-borne flaviviruses, including yellow fever virus (yfv), zika virus (zikv), and west nile virus (wnv), profoundly affect human health. the successful transmission of these viruses to a human host depends on the pathogen's ability to overcome a potentially sterilizing immune response in the vector mosquito. similar to other invertebrate animals and plants, the mosquito's rna silencing pathway comprises its primary antiviral defense. although a diverse range of plant and insect viruses has ...201627849599
peroxisomal protein pex13 functions in selective autophagy.pex13 is an integral membrane protein on the peroxisome that regulates peroxisomal matrix protein import during peroxisome biogenesis. mutations in pex13 and other peroxin proteins are associated with zellweger syndrome spectrum (zss) disorders, a subtype of peroxisome biogenesis disorder characterized by prominent neurological, hepatic, and renal abnormalities leading to neonatal death. the lack of functional peroxisomes in zss patients is widely accepted as the underlying cause of disease; how ...201627827795
deep sequencing analysis reveals the mycoviral diversity of the virome of an avirulent isolate of rhizoctonia solani ag-2-2 iv.rhizoctonia solani represents an important plant pathogenic basidiomycota species complex and the host of many different mycoviruses, as indicated by frequent detection of dsrna elements in natural populations of the fungus. to date, eight different mycoviruses have been characterized in rhizoctonia and some of them have been reported to modulate its virulence. dsrna extracts of the avirulent r. solani isolate dc17 (ag2-2-iv) displayed a diverse pattern, indicating multiple infections with mycov ...201627814394
genome investigations of vector competence in aedes aegypti to inform novel arbovirus disease control approaches.dengue (denv), yellow fever, chikungunya, and zika virus transmission to humans by a mosquito host is confounded by both intrinsic and extrinsic variables. besides virulence factors of the individual arboviruses, likelihood of virus transmission is subject to variability in the genome of the primary mosquito vector, aedes aegypti. the "vectorial capacity" of a. aegypti varies depending upon its density, biting rate, and survival rate, as well as its intrinsic ability to acquire, host and transmi ...201627809220
acyclic nucleoside phosphonates containing the amide study the influence of a linker rigidity and donor-acceptor properties, the p-ch2-o-chr- fragment in acyclic nucleoside phosphonates (e.g., acyclovir, tenofovir) was replaced by the p-ch2-hn-c(o)- residue. the respective phosphonates were synthesized in good yields by coupling the straight chain of ω-aminophosphonates and nucleobase-derived acetic acids with edc. based on the (1)h and (13)c nmr data, the unrestricted rotation within the methylene and 1,2-ethylidene linkers in phosphonates fro ...201627881885
crystal structure of glycoprotein c from a hantavirus in the post-fusion conformation.hantaviruses are important emerging human pathogens and are the causative agents of serious diseases in humans with high mortality rates. like other members in the bunyaviridae family their m segment encodes two glycoproteins, gn and gc, which are responsible for the early events of infection. hantaviruses deliver their tripartite genome into the cytoplasm by fusion of the viral and endosomal membranes in response to the reduced ph of the endosome. unlike phleboviruses (e.g. rift valley fever vi ...201627783673
mutagenic effects of ribavirin on hepatitis e virus-viral extinction versus selection of fitness-enhancing mutations.hepatitis e virus (hev), an important agent of viral hepatitis worldwide, can cause severe courses of infection in pregnant women and immunosuppressed patients. to date, hev infections can only be treated with ribavirin (rbv). major drawbacks of this therapy are that rbv is not approved for administration to pregnant women and that the virus can acquire mutations, which render the intra-host population less sensitive or even resistant to rbv. one of the proposed modes of action of rbv is a direc ...201627754363
interferon gamma: influence on neural stem cell function in neurodegenerative and neuroinflammatory disease.interferon-gamma (ifnγ), a pleiotropic cytokine, is expressed in diverse neurodegenerative and neuroinflammatory conditions. its protective mechanisms are well documented during viral infections in the brain, where ifnγ mediates non-cytolytic viral control in infected neurons. however, ifnγ also plays both protective and pathological roles in other central nervous system (cns) diseases. of the many neural cells that respond to ifnγ, neural stem/progenitor cells (nspcs), the only pluripotent cell ...201627774000
amyloid-β effects on synapses and memory require ampa receptor subunit glua3.amyloid-β (aβ) is a prime suspect for causing cognitive deficits during the early phases of alzheimer's disease (ad). experiments in ad mouse models have shown that soluble oligomeric clusters of aβ degrade synapses and impair memory formation. we show that all aβ-driven effects measured in these mice depend on ampa receptor (ampar) subunit glua3. hippocampal neurons that lack glua3 were resistant against aβ-mediated synaptic depression and spine loss. in addition, aβ oligomers blocked long-term ...201627708157
high insecticides resistance in culex pipiens (diptera: culicidae) from tehran, capital of iran.during recent years transmission of dirofilaria immitis (dog heart worm) by culex pipiens and west nile virus have been reported from iran. the present study was preformed for evaluating the susceptibility status of cx. pipiens collected from capital city of tehran, iran.201628032100
climate change influences on the global potential distribution of the mosquito culex quinquefasciatus, vector of west nile virus and lymphatic filariasis.rapid emergence of most vector-borne diseases (vbds) may be associated with range expansion of vector populations. culex quinquefasciatus say 1823 is a potential vector of west nile virus, saint louis encephalitis virus, and lymphatic filariasis. we estimated the potential distribution of cx. quinquefasciatus under both current and future climate conditions. the present potential distribution of cx. quinquefasciatus showed high suitability across low-latitude parts of the world, reflecting the c ...201627695107
a 3'-end structure in rna2 of a crinivirus is essential for viral rna synthesis and contributes to replication-associated translation activity.the terminal ends in the genome of rna viruses contain features that regulate viral replication and/or translation. we have identified a y-shaped structure (yss) in the 3' terminal regions of the bipartite genome of lettuce chlorosis virus (lcv), a member in the genus crinivirus (family closteroviridae). the yss is the first in this family of viruses to be determined using selective 2'-hydroxyl acylation analyzed by primer extension (shape). using luciferase constructs/replicons, in vivo and in ...201627694962
differential expression pattern of vago in bumblebee (bombus terrestris), induced by virulent and avirulent virus infections.viruses are one of the main drivers of the decline of domesticated and wild bees but the mechanisms of antiviral immunity in pollinators are poorly understood. recent work has suggested that next to the small interfering rna (sirna) pathway other immune-related pathways play a role in the defense of the bee hosts against viral infection. in addition, vago plays a role in the cross-talk between the innate immune pathways in culex mosquito cells. here we describe the vago orthologue in bumblebees ...201627680717
quantification of virus particles using nanopore-based resistive-pulse sensing techniques.viruses have drawn much attention in recent years due to increased recognition of their important roles in virology, immunology, clinical diagnosis, and therapy. because the biological and physical properties of viruses significantly impact their applications, quantitative detection of individual virus particles has become a critical issue. however, due to various inherent limitations of conventional enumeration techniques such as infectious titer assays, immunological assays, and electron micro ...201627713738
morphological abnormalities and cell death in the asian citrus psyllid (diaphorina citri) midgut associated with candidatus liberibacter asiaticus.candidatus liberibacter asiaticus (clas) is a phloem-limited, gram-negative, fastidious bacterium that is associated with the development of citrus greening disease, also known as huanglongbing (hlb). clas is transmitted by the asian citrus psyllid (acp) diaphorina citri, in a circulative manner. two major barriers to transmission within the insect are the midgut and the salivary glands. we performed a thorough microscopic analysis within the insect midgut following exposure to clas-infected cit ...201627630042
amphotericin b inhibits enterovirus 71 replication by impeding viral entry.enterovirus 71 (ev71) infection causes hand-foot-and-mouth disease that leads to cardiopulmonary complications and death in young children. there is thus an urgent need to find new treatments to control ev71 infection. in this study, we report potent inhibition of ev71 by a polyene antibiotic amphotericin b. amphotericin b profoundly diminished the expression of ev71 rna and viral proteins in the rd cells and the hek293 cells. as a result, ev71 production was inhibited by amphotericin b with an ...201627608771
m2bp inhibits hiv-1 virion production in a vimentin filaments-dependent manner.m2bp (also called 90k) is an interferon-stimulated gene product that is upregulated in hiv-1 infection. a recent study revealed that m2bp reduces the infectivity of hiv-1 by inhibiting the processing of the viral envelope protein. here we report that in addition to reducing viral infectivity, m2bp inhibits hiv-1 virion production. we provide evidence showing that m2bp inhibits hiv-1 gag trafficking to the plasma membrane in a vimentin-dependent manner. when vimentin filaments were collapsed by t ...201627604950
global profiling of the cellular alternative rna splicing landscape during virus-host interactions.alternative splicing (as) is a central mechanism of genetic regulation which modifies the sequence of rna transcripts in higher eukaryotes. as has been shown to increase both the variability and diversity of the cellular proteome by changing the composition of resulting proteins through differential choice of exons to be included in mature mrnas. in the present study, alterations to the global rna splicing landscape of cellular genes upon viral infection were investigated using mammalian reoviru ...201627598998
excess cerebral tnf causing glutamate excitotoxicity rationalizes treatment of neurodegenerative diseases and neurogenic pain by anti-tnf agents.the basic mechanism of the major neurodegenerative diseases, including neurogenic pain, needs to be agreed upon before rational treatments can be determined, but this knowledge is still in a state of flux. most have agreed for decades that these disease states, both infectious and non-infectious, share arguments incriminating excitotoxicity induced by excessive extracellular cerebral glutamate. excess cerebral levels of tumor necrosis factor (tnf) are also documented in the same group of disease ...201627596607
trim14 inhibits hepatitis c virus infection by spry domain-dependent targeted degradation of the viral ns5a protein.tripartite motif 14 (trim14) was reported to function as a mitochondrial signaling adaptor in mediating innate immune responses. however, the involvement of trim14 in host defense against viral infection and molecular mechanisms remain unclear. here, we demonstrated that enforced expression of trim14 could potently inhibit the infection and replication of hcv in hepatocytes, whereas trim14 knockout cells became more susceptible to hcv infection. interestingly, further experiments revealed that s ...201627578425
consecutive inhibition of isg15 expression and isgylation by cytomegalovirus regulators.interferon-stimulated gene 15 (isg15) encodes an ubiquitin-like protein that covalently conjugates protein. protein modification by isg15 (isgylation) is known to inhibit the replication of many viruses. however, studies on the viral targets and viral strategies to regulate isgylation-mediated antiviral responses are limited. in this study, we show that human cytomegalovirus (hcmv) replication is inhibited by isgylation, but the virus has evolved multiple countermeasures. hcmv-induced isg15 expr ...201627564865
type-1 angiotensin receptor signaling in central nervous system myeloid cells is pathogenic during fatal alphavirus encephalitis in mice.alphaviruses can cause fatal encephalitis in humans. natural infections occur via the bite of infected mosquitos, but aerosol transmissibility makes some of these viruses potential bioterrorism agents. central nervous system (cns) host responses contribute to alphavirus pathogenesis in experimental models and are logical therapeutic targets. we investigated whether reactive oxygen species (ros) generated by nicotinamide adenine dinucleotide phosphate (nadph) oxidase (nox) activity within the cns ...201627562117
working with zika and usutu viruses in vitro.usutu (usuv) and zika (zikv) viruses are emerging arboviruses of significant medical and veterinary importance. these viruses have not been studied as well as other medically important arboviruses such as west nile (wnv), dengue (denv), or chikungunya (chikv) viruses. as such, information regarding the behavior of zikv and usuv viruses in the laboratory is dated. usutu virus re-emerged in austria in 2001 and has since spread throughout the european and asian continents causing significant mortal ...201627541001
functional translational readthrough: a systems biology perspective.translational readthrough (tr) has come into renewed focus because systems biology approaches have identified the first human genes undergoing functional translational readthrough (ftr). ftr creates functional extensions to proteins by continuing translation of the mrna downstream of the stop codon. here we review recent developments in tr research with a focus on the identification of ftr in humans and the systems biology methods that have spurred these discoveries.201627490485
cytoplasmic drosha activity generated by alternative splicing.rnase iii enzyme drosha interacts with dgcr8 to form the microprocessor, initiating canonical microrna (mirna) maturation in the nucleus. here, we re-evaluated where drosha functions in cells using drosha and/or dgcr8 knock out (ko) cells and cleavage reporters. interestingly, a truncated drosha mutant located exclusively in the cytoplasm cleaved pri-mirna effectively in a dgcr8-dependent manner. in addition, we demonstrated that in vitro generated pri-mirnas when transfected into cells could be ...201627471035
the role of phlebovirus glycoproteins in viral entry, assembly and release.bunyaviruses are enveloped viruses with a tripartite rna genome that can pose a serious threat to animal and human health. members of the phlebovirus genus of the family bunyaviridae are transmitted by mosquitos and ticks to humans and include highly pathogenic agents like rift valley fever virus (rvfv) and severe fever with thrombocytopenia syndrome virus (sftsv) as well as viruses that do not cause disease in humans, like uukuniemi virus (uukv). phleboviruses and other bunyaviruses use their e ...201627455305
exaptive origins of regulated mrna decay in eukaryotes.eukaryotic gene expression is extensively controlled at the level of mrna stability and the mechanisms underlying this regulation are markedly different from their archaeal and bacterial counterparts. we propose that two such mechanisms, nonsense-mediated decay (nmd) and motif-specific transcript destabilization by ccch-type zinc finger rna-binding proteins, originated as a part of cellular defense against rna pathogens. these branches of the mrna turnover pathway might have been used by primeva ...201627438915
n-terminal domain of feline calicivirus (fcv) proteinase-polymerase contributes to the inhibition of host cell transcription.feline calicivirus (fcv) infection results in the inhibition of host protein synthesis, known as "shut-off". however, the precise mechanism of shut-off remains unknown. here, we found that the fcv strain 2280 proteinase-polymerase (pp) protein can suppress luciferase reporter gene expression driven by endogenous and exogenous promoters. furthermore, we found that the n-terminal 263 aa of pp (ppn-263) determined its shut-off activity using the expression of truncated proteins. however, the same d ...201627447663
dissecting virus infectious cycles by cryo-electron microscopy. 201627362353
viral inhibition of the ifn-induced jak/stat signalling pathway: development of live attenuated vaccines by mutation of viral-encoded ifn-antagonists.the interferon (ifn) induced anti-viral response is amongst the earliest and most potent of the innate responses to fight viral infection. the induction of the janus kinase/signal transducer and activation of transcription (jak/stat) signalling pathway by ifns leads to the upregulation of hundreds of interferon stimulated genes (isgs) for which, many have the ability to rapidly kill viruses within infected cells. during the long course of evolution, viruses have evolved an extraordinary range of ...201627367734
tnfα impairs rhabdoviral clearance by inhibiting the host autophagic antiviral response.tnfα is a pleiotropic pro-inflammatory cytokine with a key role in the activation of the immune system to fight viral infections. despite its antiviral role, a few viruses might utilize the host produced tnfα to their benefit. some recent reports have shown that anti-tnfα therapies could be utilized to treat certain viral infections. however, the underlying mechanisms by which tnfα can favor virus replication have not been identified. here, a rhabdoviral infection model in zebrafish allowed us t ...201627351838
the strica homolog aacasps16 is involved in apoptosis in the yellow fever vector, aedes albopictus.caspases are a family of cysteine proteases playing essential roles during apoptosis. seven caspases identified in drosophila were dronc, dredd, strica, dcp-1, decay, drice and damm. among them, strica is an insect-specific caspase containing a long serine- and threonine- rich prodomain, of which function is not yet well studied. here we identified a homolog of strica from aedes albopictus, named as aacasps16. aacasps16 encoded a protein containing a putative serine- and threonine-rich prodomain ...201627351972
suppression of kaposi's sarcoma-associated herpesvirus infection and replication by 5'-amp-activated protein kinase.the host intracellular antiviral restriction factors inhibit viral infection and replication. the 5'-amp-activated protein kinase (ampk) is a cellular energy sensor regulating metabolic homeostasis. activated ampk inhibits the replication of numerous rna viruses but enhances the entry of vaccinia virus. however, the role of ampk in herpesvirus infection is unclear. in this study, we showed that the constitutive ampk activity restricted kaposi's sarcoma-associated herpesvirus (kshv) lytic replica ...201627147746
the emerging duck flavivirus is not pathogenic for primates and is highly sensitive to mammalian interferon antiviral signaling.flaviviruses pose a significant threat to both animals and humans. recently, a novel flavivirus, duck tembusu virus (dtmuv), was identified to be the causative agent of a serious duck viral disease in asia. its rapid spread, expanding host range, and uncertain transmission routes have raised substantial concerns regarding its potential threats to nonavian hosts, including humans. here, we demonstrate that dtmuv is not pathogenic for nonhuman primates and is highly sensitive to mammal type i inte ...201627147750
recent advances in lentiviral vaccines for hiv-1 infection.the development of an effective hiv vaccine to prevent and/or cure hiv remains a global health priority. given their central role in the initiation of adaptive immune responses, dendritic cell (dc)-based vaccines are being increasingly explored as immunotherapeutic strategies to enhance hiv-specific t cells in infected individuals and, thus, promote immune responses that may help facilitate a functional cure. hiv-1-based lentiviral (lv) vectors have inherent advantages as dc vaccine vectors due ...201627446074
mosquito saliva increases endothelial permeability in the skin, immune cell migration, and dengue pathogenesis during antibody-dependent enhancement.dengue remains the most prevalent arthropod-borne viral disease in humans. while probing for blood vessels, aedes aegypti and ae. albopictus mosquitoes transmit the four serotypes of dengue virus (denv1-4) by injecting virus-containing saliva into the skin. even though arthropod saliva is known to facilitate transmission and modulate host responses to other pathogens, the full impact of mosquito saliva on dengue pathogenesis is still not well understood. inoculating mice lacking the interferon-α ...201627310141
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