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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
inactivation of viruses in platelet and plasma products using a riboflavin-and-uv-based photochemical treatment.multilayered blood safety programs reduce the risk of transfusion-transmitted diseases; however, there remains a risk of window period transmission of screened viruses and transmission of unscreened and emerging viruses from asymptomatic donors. to reduce this risk, a riboflavin-and-uv-light-based pathogen reduction process was evaluated against eight viral agents.201525731607
phosphorylation of eif2α is responsible for the failure of the picornavirus internal ribosome entry site to direct translation from sindbis virus replicons.translation directed by the poliovirus (pv) or encephalomyocarditis virus (emcv) internal ribosome entry site (ires) is very inefficient when expressed from sindbis virus (sv) replicons. this inhibition can be rescued by co-expression of pv 2a protease (2a(pro)). inhibition correlates with the extensive phosphorylation of eukaryotic initiation factor (eif) 2α induced by sv replication. confirmation that pv or emcv ires-driven translation can function when eif2α is not phosphorylated was obtained ...201323255624
antiviral activities of whey proteins.milk contains an array of proteins with useful bioactivities. many milk proteins encompassing native or chemically modified casein, lactoferrin, alpha-lactalbumin, and beta-lactoglobulin demonstrated antiviral activities. casein and alpha-lactalbumin gained anti-hiv activity after modification with 3-hydroxyphthalic anhydride. many milk proteins inhibited hiv reverse transcriptase. bovine glycolactin, angiogenin-1, lactogenin, casein, alpha-lactalbumin, beta-lactoglobulin, bovine lactoferrampin, ...201526198883
translation of viral mrnas that do not require eif4e is blocked by the inhibitor 4egi-1.high throughput screening has rendered new inhibitors of eukaryotic protein synthesis. one such molecule, 4egi-1 has been reported to selectively block the initiation factor eif4e. we have investigated the action of this inhibitor on translation directed by several viral mrnas which, in principle, do not utilize eif4e. we found that 4egi-1 inhibits translation directed by poliovirus ires, in rabbit reticulocyte lysates, to a similar extent as capped mrna. moreover, 4egi-1 inhibits translation dr ...201323870416
antiviral activity of a derivative of the photosensitive compound hypericin.eight synthetic compounds related to the photosensitive antiviral quinonic plant compound hypericin were screened for light-mediated antiviral activity. the compound 2,5,9,12-tetra(carboxyethylthiomethyl) hypericin was found to have activity against membrane-enveloped sindbis virus and murine cytomegalovirus but not against the non-enveloped poliovirus. the activity of this compound was investigated in more detail. the mechanism of action appeared to be of the photosensitive singlet oxygen type ...199623194968
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
triple mammalian/yeast/bacterial shuttle vectors for single and combined lentivirus- and sindbis virus-mediated infections of neurons.today, a large variety of viral vectors is available for ectopic gene expression in mammalian cell cultures or in vivo. among them, infection with sindbis virus- or lentivirus-derived constructs is often used to address biological questions or for applications in neuronal therapies. however, cloning of genes of interest is time consuming, since it relies on restriction and ligation, frequently of pcr-generated dna fragments with suitable restriction sites introduced by the primers employed. we h ...201222327349
sindbis virus replicase-based dna vaccine construct encoding fmdv-specific multivalent epitope gene: studies on its immune responses in guinea pigs.foot-and-mouth disease (fmd) is still a perennial global menace affecting livestock health and production. it is imperative to figure out new ways to curb this disease. in this study, a sindbis virus replicase-based dna vaccine, psincmv-vac-meg990, encoding a multivalent epitope gene (representing tandemly linked vp1 c-terminal halves of three foot-and-mouth disease virus (fmdv) serotypes) was constructed. in vitro transfection studies in bhk-21 cells revealed that the construct was able to expr ...201222702835
synthesis and antiviral evaluation of c5-substituted-(1,3-diyne)-2'-deoxyuridines.starting from acetylated 5-ethynyl-2'-deoxyuridine (3), 14 hitherto unknown c5-substituted-(1,3-diyne)-2'-deoxyuridines (with cyclopropyl, hydroxymethyl, methylcyclopentane, p-(substituted)phenyl and disubstituted-phenyl substituents) have been synthesized via a nickel-copper catalyzed c-h activation between two terminal alkynes, in yields ranging from 19% to 67%. their antiviral activities were measured against a large number of dna and rna viruses including herpes simplex virus type 1 and type ...201222578783
membrane targeting of an alpha-like tetravirus replicase is directed by a region within the rna-dependent rna polymerase domain.the members of the family tetraviridae are small positive-sense insect rna viruses that exhibit stringent host specificity and a high degree of tissue tropism, suggesting that complex virus-host interactions are likely to occur during infection and viral replication. the alpha-like replicase of helicoverpa armigera stunt virus (hasv) (genus omegatetravirus) has been proposed to associate with membranes of the endocytic pathway, which is similar to semliki forest virus, sindbis virus and rubella ...201222535773
[updated inventory of mosquitoes (diptera: culicidae) of the island of la réunion, indian ocean].a literature analysis coupled with new entomological surveys conducted between 2009 and 2012 led to changes in the list of mosquito species present on the island of la réunion. using morphological criteria, orthopodomyia arboricollis is replaced by or. reunionensis. on the basis of morphometrical and genetic criteria, culex univittatus is replaced by cx. neavei. cx. poicilipes, which was already reported missing 40 years ago, has not been found again. anopheles arabiensis is confirmed as the onl ...201323681758
[complete genome characterization of the kyzylagach virus (kyzv) (togaviridae, alphavirus, sindbis serogroup) isolated from mosquitoes culex modestus ficalbi, 1889 (culicinae) collected in a colony of herons (ardeidae leach, 1820) in azerbaijan].complete genome sequencing of the kyzylagach virus (kyzv) leiv-65a strain isolated from the culex modestus ficalbi, 1889 (culicinae), which was collected in the colony of the ardeidae leach, 1820 birds on the coast of caspian sea, kyzyl-agach bay, in the southern part of azerbaijan, was carried out. kyzv has high homology (about 99%) with the chinese xj-160 strain of the sindbis virus (sinv) isolated from anopheles sp. in xinjiang uyghur autonomic region (north-eastern china). homologies of kyzv ...201425895208
effect of mixed infections of sindbis and la crosse viruses on replication of each virus in vitro.mixed viral infections are ubiquitous in natural populations and may have significant but unpredictable biological and epidemiological consequences. these infections may be acquired simultaneously (coinfection) or result from two single infections at different time intervals (superinfection). this study investigated the effect of mixed infections of two mosquito-borne viruses, sindbis (sinv) and la crosse (lacv), on the replication of each virus in vertebrate baby hamster kidney (bhk) and invert ...201424463260
[the concept of emerging viral diseases: what risk for reunion island?].in reunion island, the risk of emerging infectious diseases lies mainly in several viral zoonoses: west nile fever, sindbis virus, nipah virus, wesselsbron virus, rift valley fever and japanese encephalitis. there morbidity and consequences are more or less important but they all have a non-negligible epidemic potential, so they have to be monitored. indeed, the struggle against these emerging infectious diseases requires an early detection of the cases, thus a surveillance system capable of det ...201323765703
enveloped virus flocculation and removal in osmolyte solutions.our ability to reduce infectious disease burden throughout the world has been greatly improved by the creation of vaccines. however, worldwide immunization rates are low. the two most likely reasons are the lack of sufficient distribution in underdeveloped countries and the high cost of vaccine products. the high costs are due to the difficulties of manufacturing individual vaccine products with specialized purification trains. in this study, we propose to use virus flocculation in osmolytes, fo ...201525865274
virus adsorption of water-stable quaternized chitosan nanofibers.the burden of unsafe drinking water is responsible for millions of deaths each year. to relieve this burden, we are in search of an inexpensive material that can adsorb pathogens from drinking water. in this pursuit, we have studied the natural carbohydrate, chitosan. to impart virus removal features, chitosan has been functionalized with a quaternary amine to form quaternized chitosan n-[(2-hydroxyl-3-trimethylammonium) propyl] chitosan (htcc). htcc can be electrospun into nanofibers with the n ...201424561959
in vivo administration of recombinant alphavirus into rodents.the alphaviruses semliki forest virus (sfv) and sindbis virus (sin) have been used frequently as expression vectors in vitro and in vivo. usually, these systems consist of replication-deficient vectors that require a helper vector for packaging of recombinant particles. replication-proficient vectors have also been engineered. alphaviral vectors can be used as nucleic-acid-based vectors (dna and rna) or infectious particles. high-titer viral production is achieved in <2 d. the broad host range o ...201222854568
delivery of recombinant alphavirus into hippocampal slice tissue culture.the alphaviruses semliki forest virus (sfv) and sindbis virus (sin) have been used frequently as expression vectors in vitro and in vivo. usually, these systems consist of replication-deficient vectors that require a helper vector for packaging of recombinant particles. replication-proficient vectors have also been engineered. alphaviral vectors can be used as nucleic-acid-based vectors (dna and rna) or infectious particles. high-titer viral production is achieved in <2 d. the broad host range o ...201222854567
generation of recombinant alphaviral vectors.the alphaviruses semliki forest virus and sindbis virus have been used frequently as expression vectors in vitro and in vivo. usually, these systems consist of replication-deficient vectors that require a helper vector for packaging of recombinant particles. replication-proficient vectors have also been engineered. alphaviral vectors can be used as nucleic-acid-based vectors (dna and rna) or infectious particles. high-titer viral production is achieved in <2 d. the broad host range of alphavirus ...201222753600
human cases of sindbis fever in south africa, 2006-2010.sindbis virus (sinv), the prototype positive-sense rna alphavirus, causes febrile arthritis and is present throughout afro-eurasia. little is known of the epidemiology of sindbis fever due to insufficient surveillance in most endemic countries. the epidemiological features of sindbis fever in humans in south africa are described here based on a retrospective study of suspected arbovirus cases submitted for laboratory investigation from 2006 to 2010. cases were detected annually mostly during the ...201423611492
neurotropic alphaviruses can propagate without capsid. 201728129638
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
semliki forest virus and sindbis virus, but not vaccinia virus, require glycolysis for optimal replication.viruses are obligate intracellular pathogens which rely on the cell's machinery to produce the energy and macromolecules required for replication. infection is associated with a modified metabolic profile and one pathway which can be modified is glycolysis. in this study, we investigated if the glycolysis pathway is required for alphavirus replication. pre-treatment of vero cells with three different glycolysis inhibitors (2-deoxyglucose, lonidamine and oxamate) resulted in a significant reducti ...201526297236
determination of alphaviral titers.the alphaviruses semliki forest virus and sindbis virus have been used frequently as expression vectors in vitro and in vivo. usually, these systems consist of replication-deficient vectors that require a helper vector for packaging of recombinant particles. replication-proficient vectors have also been engineered. alphaviral vectors can be used as nucleic-acid-based vectors (dna and rna) or infectious particles. the broad host range of alphaviruses facilitates studies in mammalian and nonmammal ...201222753602
purification and concentration of alphavirus.the alphaviruses semliki forest virus and sindbis virus have been used frequently as expression vectors in vitro and in vivo. usually, these systems consist of replication-deficient vectors that require a helper vector for packaging of recombinant particles. replication-proficient vectors have also been engineered. alphaviral vectors can be used as nucleic-acid-based vectors (dna and rna) or infectious particles. high-titer viral production is achieved in <2 d. the broad host range of alphavirus ...201222753601
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
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
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
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
t cell-derived interleukin-10 is an important regulator of the th17 response during lethal alphavirus encephalomyelitis.neuroadapted sindbis virus infection of mice causes t cell-mediated fatal encephalomyelitis. in the absence of il-10, pathogenic th17 cells are increased and disease is accelerated. lymphoid and myeloid cell contributions to il-10 production were determined using vertx il-10 transcriptional egfp reporter mice. effector and regulatory cd4(+) and cd8(+) t cells in the brain, but not the cervical lymph nodes, were the primary producers of il-10. th17 and th1/th17 cells were increased in mice that l ...201627235350
alphavirus encephalomyelitis: mechanisms and approaches to prevention of neuronal damage.mosquito-borne viruses are important causes of death and long-term neurologic disability due to encephalomyelitis. studies of mice infected with the alphavirus sindbis virus have shown that outcome is dependent on the age and genetic background of the mouse and virulence of the infecting virus. age-dependent susceptibility reflects the acquisition by neurons of resistance to virus replication and virus-induced cell death with maturation. in mature mice, the populations of neurons most susceptibl ...201627114366
sindbis virus as a human pathogen-epidemiology, clinical picture and pathogenesis.sindbis virus (sinv; family togaviridae, genus alphavirus) is an enveloped rna virus widely distributed in eurasia, africa, oceania and australia. sinv is transmitted among its natural bird hosts via mosquitoes. human disease caused by sinv infection has been reported mainly in south africa and in northern europe. vector mosquito abundance affects the annual incidence of sinv infections with occasional outbreaks of up to 1500 patients. symptoms include fever, malaise, rash and musculoskeletal pa ...201626990827
concentration of sindbis virus with optimized gradient insulator-based dielectrophoresis.biotechnology, separation science, and clinical research are impacted by microfluidic devices. separation and manipulation of bioparticles such as dna, protein and viruses are performed on these platforms. microfluidic systems provide many attractive features, including small sample size, rapid detection, high sensitivity and short processing time. dielectrophoresis (dep) and electrophoresis are especially well suited to microscale bioparticle control and have been demonstrated in many formats. ...201626878279
synthesis and antiviral properties of novel indole-based thiosemicarbazides and 4-thiazolidinones.a novel series of indolylthiosemicarbazides (6a-6g) and their cyclization products, 4-thiazolidinones (7a-7g), have been designed, synthesized and evaluated, in vitro, for their antiviral activity against a wide range of dna and rna viruses. compounds 6a, 6b, 6c and 6d exhibited notable antiviral activity against coxsackie b4 virus, at ec50 values ranging from 0.4 to 2.1 μg/ml. the selectivity index (ratio of cytotoxic to antivirally effective concentration) values of these compounds were betwee ...201626707844
oncolytic viral therapy for neuroblastoma cells with sindbis virus ar339 strain.with current treatment regimens, high-risk neuroblastoma (nb) remains largely incurable. oncolytic viral therapy uses replication-competent viruses, like sindbis virus (sinv), to kill cancers. the sinv ar339 strain is blood borne and relatively non-virulent. we evaluated the feasibility of sinv ar339 for treating human nb.201526298056
differential action of pateamine a on translation of genomic and subgenomic mrnas from sindbis virus.pateamine a (pat a) is a natural marine product that interacts specifically with the translation initiation factor eif4a leading to the disruption of the eif4f complex. in the present study, we have examined the activity of pat a on the translation of sindbis virus (sinv) mrnas. translation of genomic mrna is strongly suppressed by pat a, as shown by the reduction of nsp1 or nsp2 synthesis. notably, protein synthesis directed by subgenomic mrna is resistant to pat a inhibition when the compound ...201526057151
trapping biases of culex torrentium and culex pipiens revealed by comparison of captures in cdc traps, ovitraps, and gravid traps.we evaluate three trapping methods for their effectiveness at capturing culex pipiens and culex torrentium, both enzootic vectors of bird-associated viruses in europe. the comparisons, performed in two regions in sweden, were among cdc traps baited with carbon dioxide, gravid traps, and ovitraps baited with hay infusion. the proportions of the two culex species in a catch differed between trap types, with cdc traps catching a lower proportion of cx. torrentium than both gravid traps and ovitraps ...201526047196
alteration of cell cycle progression by sindbis virus infection.we examined the impact of sindbis virus (sinv) infection on cell cycle progression in a cancer cell line, hela, and a non-cancerous cell line, vero. cell cycle analyses showed that sinv infection is able to alter the cell cycle progression in both hela and vero cells, but differently, especially during the early stage of infection. sinv infection affected the expression of several cell cycle regulators (cdk4, cdk6, cyclin e, p21, cyclin a and cyclin b) in hela cells and caused hela cells to accu ...201525976675
enhanced expression of trim14 gene suppressed sindbis virus reproduction and modulated the transcription of a large number of genes of innate immunity.in the present research, we have studied an influence of enhanced expression trim14 on alphavirus sindbis (sinv, togaviridae family) infection. in the hek293 cells transfected with human trim14 gene (hek-trim14), sinv yield after infection was decreased 1000-10,000 times (3-4 lg of tcd50/ml) at 24 h p.i. and considerably less (1-2 lg of tcd50/ml) at 48 h p.i. analysis of the expression of 43 genes directly or indirectly involved in innate immune machine in hek-trim14 non-infected cells comparing ...201525948474
antiviral and antitumor activities of the lectin extracted from aspidistra elatior.lectins, a group of highly diverse proteins of non-immune origin and are ubiquitously distributed in plants, animals and fungi, have multiple significant biological functions, such as anti-fungal, anti-viral and, most notably, anti-tumor activities. a lectin was purified from the rhizomes of aspidistra elatior blume, named a. elatior lectin (ael). in vitro experiments showed that the minimum inhibitory concentrations of ael against the vesicular stomatitis virus, coxsackie virus b4, and respirat ...201525854839
different cortical projections from three subdivisions of the rat lateral posterior thalamic nucleus: a single-neuron tracing study with viral vectors.the lateral posterior thalamic nucleus (lp) is one of the components of the extrageniculate pathway in the rat visual system, and is cytoarchitecturally divided into three subdivisions--lateral (lpl), rostromedial (lprm), and caudomedial (lpcm) portions. to clarify the differences in the dendritic fields and axonal arborisations among the three subdivisions, we applied a single-neuron labeling technique with viral vectors to lp neurons. the proximal dendrites of lpl neurons were more numerous th ...201525832313
detection and isolation of sindbis virus from mosquitoes captured during an outbreak in sweden, 2013.mosquito-borne alphaviruses have the potential to cause large outbreaks throughout the world. here we investigated the causative agent of an unexpected sindbis virus (sinv) outbreak during august-september, 2013, in a previously nonendemic region of sweden. mosquitoes were collected using carbon dioxide-baited cdc traps at locations close to human cases. the mosquitoes were initially screened as large pools by sinv-specific quantitative rt-pcr, and the sinv-positive mosquitoes were species deter ...201525700044
a substitution in nsp1 combined with a double substitution in e2 glycoprotein renders sindbis-like virus xj-160 fully neurovirulent for adult mice.sindbis virus (sv) can be rendered neurovirulent for adult mice by a double substitution within the e2 glycoprotein, including replacing gln at position 55 of e2 with a his (e2-55: gln-his) and e2-70: glu to lys. however, the mutant sindbis-like virus xj-160 with the double substitution (br-e5570) does not show neurovirulence for adult mice, although the mutant apparently reduced the average survival time of neonatal mice. to produce an xj-160 virus neurovirulent for adult mice, the br-e5570 vir ...201525449361
parp13 regulates cellular mrna post-transcriptionally and functions as a pro-apoptotic factor by destabilizing trailr4 transcript.poly(adp-ribose) polymerase-13 (parp13/zap/zc3hav1) is an antiviral factor, active against specific rna viruses such as murine leukaemia virus, sindbis virus and human immunodeficiency virus. during infection, parp13 binds viral rna via its four ccch-type zinc-finger domains and targets it for degradation by recruiting cellular messenger rna (mrna) decay factors such as the exosome complex and xrn1. here we show that parp13 binds to and regulates cellular mrnas in the absence of viral infection. ...201425382312
inhibition of host protein synthesis by sindbis virus: correlation with viral rna replication and release of nuclear proteins to the cytoplasm.infection of mammalian cells by sindbis virus (sinv) profoundly blocks cellular mrna translation. experimental evidence points to viral non-structural proteins (nsps), in particular nsp2, as the mediator of this inhibition. however, individual expression of nsp1, nsp2, nsp3 or nsp1-4 does not block cellular protein synthesis in bhk cells. trans-complementation of a defective sinv replicon lacking most of the coding region for nsps by the co-expression of nsp1-4 propitiates viral rna replication ...201525329362
model-driven engineering of gene expression from rna replicons.rna replicons are an emerging platform for engineering synthetic biological systems. replicons self-amplify, can provide persistent high-level expression of proteins even from a small initial dose, and, unlike dna vectors, pose minimal risk of chromosomal integration. however, no quantitative model sufficient for engineering levels of protein expression from such replicon systems currently exists. here, we aim to enable the engineering of multigene expression from more than one species of replic ...201524877739
isolation of sindbis virus from a hooded crow in germany.sindbis virus (sinv) is an arbovirus that causes clinical symptoms, including arthritis, rash, and fever during acute human infections. in europe, sinv outbreaks are largely restricted to northern europe. intrigued by the isolation of sinv from mosquitoes in southwestern germany in 2009, we initiated a passive arbovirus-monitoring program in birds and analyzed a total of 685 samples. by this approach, we were able to detect a sinv in a hooded crow in germany for the first time. it was possible t ...201424575742
tissue-specific attenuation of oncolytic sindbis virus without compromised genetic stability.wild-type sindbis virus (sv) shows promise as an oncolytic agent, although potential off-target replication is a safety concern. to remove possible pathology reflecting virus replication in liver, muscle, and/or hematopoietic cells, microrna (mir)-response elements (mres) to liver-specific mir122, muscle-specific mir133a and mir206, or hematopoietic-specific mir142-3p were inserted into the sindbis viral genome. we compared the effectiveness of mres in two distinct genomic locations and found be ...201424568203
the alternate triad motif of the poly(adp-ribose) polymerase-like domain of the human zinc finger antiviral protein is essential for its antiviral activity.the human zinc finger antiviral protein (hzap) gene is spliced to yield a short (hzap-s) and a long (hzap-l) isoform. the long isoform possesses a poly(adp-ribose) polymerase (parp)-like domain in its c-terminus predicted to be inactive due to alterations in its triad motif compared with bona fide parps. using sindbis virus as prototype member of alphaviruses we confirmed that hzap-l is a more potent inhibitor of alphaviruses than hzap-s. specific small interfering rna knockdown of hzap-l but no ...201424457973
[construction and characterization of egfp reporter gene labeled sindbis virus].to construct and characterize egfp reporter gene labeled sindbis virus (sinv).201324319966
engineered alphavirus replicon vaccines based on known attenuated viral mutants show limited effects on immunogenicity.the immunogenicity of alphavirus replicon vaccines is determined by many factors including the level of antigen expression and induction of innate immune responses. characterized attenuated alphavirus mutants contain changes to the genomic 5' utr and mutations that result in altered non-structural protein cleavage timing leading to altered levels of antigen expression and interferon (ifn) induction. in an attempt to create more potent replicon vaccines, we engineered a panel of venezuelan equine ...201324210122
neuronal network imaging in acute slices using ca2+ sensitive bioluminescent reporter.genetically encoded indicators are valuable tools to study intracellular signaling cascades in real time using fluorescent or bioluminescent imaging techniques. imaging of ca(2+) indicators is widely used to record transient intracellular ca(2+) increases associated with bioelectrical activity. the natural bioluminescent ca(2+) sensor aequorin has been historically the first ca(2+) indicator used to address biological questions. aequorin imaging offers several advantages over fluorescent reporte ...201424166366
seroprevalence of sindbis virus and associated risk factors in northern sweden.mosquito-borne sindbis virus (sinv) cause disease characterized by rash, fever and arthritis which often leads to long-lasting arthralgia. to determine the seroprevalence of sinv and associated risk factors in northern sweden, a randomly selected population aged between 25 and 74 years were invited to join the monica study. serum from 1611 samples were analysed for specific igg antibodies. overall, 2·9% had igg against sinv. more men (3·7%) than women (2·0%) were sinv seropositive (p = 0·047) an ...201424029159
specific nucleotide changes in the subgenomic promoter region influence infectivity of the sindbis virus.transcription of the subgenomic mrna of sindbis virus (sinv) is initiated at a subgenomic promoter (sp). alignment of sinv sequences identified a 68-nucleotide conserved domain spanning -19 to +49 relative to the subgenomic mrna start site. nucleotide t or c is present at -18 or +49 in all known sinvs while a sindbis-like virus xj-160 has an a or t at a corresponding position. our results indicate that deletion or substitution of the t at +49 decreased the activity of sp, while substituting t fo ...201323838061
recombination of replicon and helper rnas and the emergence of propagation-competent vectors upon sindbis virus vector production.sindbis vectors have shown remarkable antitumor efficacy and tumor-targeting capacity in animal models and hold promise for cancer therapy. different packaging systems are used to produce propagation-incompetent sindbis vectors. however, the vectors produced using either dh-bb single helper rna or split helper rna can spread in permissive cell cultures. we investigated the mechanisms of vector spreading and show, here, that recombination occurs between the replicon and dh-bb helper rna, leading ...201323716190
effect of larval density and sindbis virus infection on immune responses in aedes aegypti.stressful environmental conditions during mosquito larval development may enhance susceptibility of adult mosquitoes to viral pathogens. although anti-viral defense system in mosquitoes remains uncertain, stress-related enhancement of mosquito susceptibility to viral pathogens may be due to alteration of signaling pathways such as the toll and immune deficiency (imd) pathways. to test the influence of larval density and sindbis virus (sinv) infection on mosquito toll/imd pathways, 100 or 200 aed ...201323562781
requirements for eif4a and eif2 during translation of sindbis virus subgenomic mrna in vertebrate and invertebrate host cells.we have examined the requirements for the initiation factors (eifs) eif4a and eif2 to translate sindbis virus (sv) subgenomic mrna (sgmrna) in the natural hosts of sv: vertebrate and arthropod cells. notably, this viral mrna does not utilize eif4a in sv-infected mammalian cells. however, eif4a is required to translate this mrna in transfected cells. therefore, sv sgmrna exhibits a dual mechanism for translation with respect to the use of eif4a. interestingly, sv genomic mrna requires eif4a for t ...201323189929
climatic, ecological and socioeconomic factors as predictors of sindbis virus infections in finland.mosquito-borne sindbis virus (sinv) causes rash-arthritis syndrome in finland. major outbreaks with approximately 7-year cycles have caused substantial burden of illness. forest dwelling grouse are suspected to be amplifying hosts, with the infection transmitted to humans by mosquito bites. sinv infection surveillance data for 1984–2010 were used to create a negative binomial hurdle model, with seasonality, long-term cycles, climatic, ecological and socioeconomic variables. climatic factors duri ...201323158410
phenotypic and molecular characteristics of plaque-purified mx10 virus, an oriental-australian genotype of sindbis virus from yunnan, china.a previous investigation showed that mx10 virus, recently isolated in china, belongs to the oriental-australian (o/a) genotype of sindbis virus (sinv) (wang jinglin, 2011, atmh). similar to the mre16 isolate, the prototype o/a genotype of sinv, two derivate viruses with obviously different plaque morphologies were derived from mx10 virus, which were accordingly denoted as mx10-lp and mx10-sp. mx10-lp virus exhibited higher neurovirulence in neonatal mice than mx10-sp virus. analysis of the compl ...201322965578
sindbis viral vectors target hematopoietic malignant cells.sindbis viral vectors target and inhibit the growth of various solid tumors in mouse models. however, their efficacy against blood cancer has not been well established. here, we show that sindbis vectors infect and efficiently trigger apoptosis in mouse bw5147 malignant hematopoietic t-cells, but only at low levels in human lymphoma and leukemia cells (jurkat, karpas, cem, dhl and jb). the mr 37/67 kd laminin receptor (lamr) has been suggested to be the receptor for sindbis virus. however, jb ce ...201222956041
is minocycline useful for therapy of acute viral encephalitis?minocyline is a tetracycline derivative with anti-inflammatory, anti-apoptotic, and anti-oxidant properties. therapy has proved useful in some experimental models of both noninfectious and infectious neurological diseases and also in clinical trials in humans, including acute traumatic cervical spinal cord injury. in models of viral encephalitis, treatment has shown both beneficial and deleterious effects. in reovirus infection in mice, minocycline delayed the disease, but did not improve either ...201222728256
complete coding sequence and molecular epidemiological analysis of sindbis virus isolates from mosquitoes and humans, finland.sindbis virus (sinv) is an arthropod-borne alphavirus, which causes rash-arthritis, particularly in finland. sinv is transmitted by mosquitoes in finland but thus far no virus has been isolated from mosquitoes. in this study, we report the isolation of the first sinv strain from mosquitoes in finland and its full-length protein-coding sequence. we furthermore describe the full-length coding sequence of six sinv strains previously isolated from humans in finland and from a mosquito in russia. the ...201222647374
prolonged myalgia in sindbis virus infection: case description and in vitro infection of myotubes and myoblasts.sindbis virus (sinv) is a mosquito-borne alphavirus found in eurasia, africa, and oceania. clinical sinv infection is characterized by febrile rash and arthritis and sometimes prolonged arthralgia and myalgia. the pathophysiological mechanisms of musculoskeletal and rheumatic disease caused by sinv are inadequately understood.201222615321
temperature and density-dependent effects of larval environment on aedes aegypti competence for an alphavirus.mosquito larvae experience multiple environmental stressors that may modify how subsequent adults interact with pathogens. we evaluated the effect of larval rearing temperature and intraspecific larval competition on adult mosquito immunity and vector competence for sindbis virus (sinv). aedes aegypti larvae were reared at two intraspecific densities (150 and 300 larvae) at 20° c and 30° c and the adults were fed artificially on citrated bovine blood containing 10(5) plaque forming units of sinv ...201222548549
clinical sindbis alphavirus infection is associated with hla-drb1*01 allele and production of autoantibodies.sindbis virus (sinv) is a mosquito-borne alphavirus found in eurasia, africa, and oceania. clinical sinv infection, characterized by arthropathic disease that may persist for years, is primarily reported in northern europe where the disease has considerable public health importance in endemic areas. the aim of this study was to investigate the role of genetic factors in the susceptibility and outcome of sinv infection and to elucidate the association between sinv infection and autoimmunity.201222523259
sindbis virus infection alters blood feeding responses and deet repellency in aedes aegypti (diptera: culicidae).aedes aegypti (l.) (diptera: culicidae) female mosquitoes infected systemically with sindbis virus (sinv) took longer than uninfected mosquitoes to locate and fully engorge on blood. on days 7 and 14 postexposure, blood feeding took 1.3 and 1.5 times longer in mosquitoes with a disseminated sinv infection, respectively. sinv dissemination did not affect the average weight of unfed ae. aegypti, but did result in a 10 and 12% increase in blood imbibed compared with mosquitoes without a positive si ...201222493862
altered behavioral responses of sindbis virus-infected aedes aegypti (diptera: culicidae) to deet and non-deet based insect repellents.changes in the time to first bite (tfb) and the bloodfeeding behavior of adult female aedes aegypti (l.) mosquitoes following dissemination of sindbis virus (sinv) were observed after exposure to repellents with the active ingredients (ai) deet, picaridin, 2-undecanone (2-u), and oil of lemon eucalyptus. dissemination of sinv significantly decreased (p<0.0001) the tfb of deet (15%) and picaridin (15%) by 46% and 37%, respectively. significant (p<0.0001) changes in activation, probing, and engorg ...201222289669
antimicrobial and cytotoxic activities of turbinariaconoides (j.agardh) kuetz.brown alga, turbinariaconoideswas successively extracted with n-hexane, cyclohexane, methanol and ethanol:water (1:1). the extracts were evaluated for antibacterial and antifungal activities by disc diffusion method. minimal inhibitory concentration was determined for active extracts by broth dilution method. the antiviral activity and cytotoxicity of the extracts were tested in human embryonic lung (hel) cells (herpes simplex virus-1, herpes simplex virus-2, vaccinia virus, vesicular stomatitis ...201024381606
bcl-2 inhibits apoptosis and extends recombinant protein production in cells infected with sindbis viral vectors.viruses carrying foreign genes are often used for the production of recombinant proteins in mammalian cells and other eukaryotic expression systems. though high levels of gene expression are possible using viral vectors, the host cell generally responds to the infection by inducing apoptotic cell death within several days, abruptly ending protein production. it has recently been demonstrated, however, that apoptosis can be suppressed in virally infected cells using anti-apoptotic genes, such as ...199622358927
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, china.in 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
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