PMID(sorted descending)
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
comparison of α-glucosyl hesperidin of citrus fruits and epigallocatechin gallate of green tea on the loss of rotavirus infectivity in cell culture.a number of secondary plant metabolites (e.g., flavonoids) possess antiviral/antimicrobial activity. most flavonoids, however, are difficult to study, as they are immiscible in water-based systems. the relatively new semisynthetic α-glucosyl hesperitin (gh), and the natural plant product epigallocatechin gallate (egcg) are unique among most flavonoids, as these flavonoids are highly soluble. the antiviral activity of these plant metabolites were investigated using the rotavirus as a model enteri ...201525972850
ability of the encephalitic arbovirus semliki forest virus to cross the blood-brain barrier is determined by the charge of the e2 glycoprotein.semliki forest virus (sfv) provides a well-characterized model system to study the pathogenesis of virus encephalitis. several studies have used virus derived from the molecular clone sfv4. sfv4 virus does not have the same phenotype as the closely related l10 or the prototype virus from which its molecular clone was derived. in mice, l10 generates a high-titer plasma viremia, is efficiently neuroinvasive, and produces a fatal panencephalitis, whereas low-dose sfv4 produces a low-titer viremia, ...201525972559
do we need a vaccine against chikungunya?during the last decade, the chikungunya (chikv) virus has expanded its range of activity, conquering new territories and becoming an important global health threat. in particular, the challenge represented by the recent emergence of chikv in the americas has strengthened the need of a safe and effective vaccine. although research on vaccines against chikv has been slow, a few vaccine candidates have been tested over the years. inactivated and attenuated vaccine candidates have shown promising re ...201525971340
alphaviruses in gene therapy.alphavirus vectors present an attractive approach for gene therapy applications due to the rapid and simple recombinant virus particle production and their broad range of mammalian host cell transduction. mainly three types of alphavirus vectors, namely naked rna, recombinant particles and dna/rna layered vectors, have been subjected to preclinical studies with the goal of achieving prophylactic or therapeutic efficacy, particularly in oncology. in this context, immunization with alphavirus vect ...201525961488
enhanced expression of trim14 gene suppressed sindbis virus reproduction and modulated the transcription of a large number of genes of innate 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 activity of flexibilane and tigliane diterpenoids from stillingia an effort to identify new potent and selective inhibitors of chikungunya virus and hiv-1 and hiv-2 virus replication, the endemic mascarene species stillingia lineata was investigated. lc/ms and bioassay-guided purification of the etoac leaf extract using a chikungunya virus-cell-based assay led to the isolation of six new (4-9) and three known (1-3) tonantzitlolones possessing the rare c20-flexibilane skeleton, along with tonantzitloic acid (10), a new linear diterpenoid, and three new (11, ...201525946116
adp-ribosyltransferases and poly adp-ribosylation.protein adp-ribosylation is an important posttranslational modification that plays versatile roles in multiple biological processes. adp-ribosylation is catalyzed by a group of enzymes known as adp-ribosyltransferases (arts). using nicotinamide adenine dinucleotide (nad(+)) as the donor, arts covalently link single or multiple adp-ribose moieties from nad(+) to the substrates, forming mono adp-ribosylation or poly adp-ribosylation (parylation). novel functions of arts and adpribosylation have be ...025938242
recovery of recombinant crimean congo hemorrhagic fever virus reveals a function for non-structural glycoproteins cleavage by furin.crimean congo hemorrhagic fever virus (cchfv) is a negative-strand rna virus of the family bunyaviridae (genus: nairovirus). in humans, cchfv causes fever, hemorrhage, severe thrombocytopenia, and high fatality. a major impediment in precisely determining the basis of cchfv's high pathogenicity has been the lack of methodology to produce recombinant cchfv. we developed a reverse genetics system based on transfecting plasmids into bsr-t7/5 and huh7 cells. in our system, bacteriophage t7 rna polym ...201525933376
inhibition of cav3.2 t-type calcium channels by its intracellular i-ii loop.voltage-dependent calcium channels (cav) of the t-type family (cav3.1, cav3.2, and cav3.3) are activated by low threshold membrane depolarization and contribute greatly to neuronal network excitability. enhanced t-type channel activity, especially cav3.2, contributes to disease states, including absence epilepsy. interestingly, the intracellular loop connecting domains i and ii (i-ii loop) of cav3.2 channels is implicated in the control of both surface expression and channel gating, indicating t ...201525931121
native wolbachia from aedes albopictus blocks chikungunya virus infection in cellulo.wolbachia, a widespread endosymbiont of terrestrial arthropods, can protect its host against viral and parasitic infections, a phenotype called "pathogen blocking". however, in some cases wolbachia may have no effect or even enhance pathogen infection, depending on the host-wolbachia-pathogen combination. the tiger mosquito aedes albopictus is naturally infected by two strains of wolbachia, walba and walbb, and is a competent vector for different arboviruses such as dengue virus (denv) and chiku ...201525923352
chikungunya virus (chikv) inhibitors from natural sources: a medicinal chemistry perspective.chikungunya virus (chikv) is one of the re-emerging "neglected" tropical diseases whose recent outbreak affected not only africa and south-east asia but also several parts of america and europe. to date, despite its serious nature, no antivirals or vaccines were developed in order to counter this resurgent infectious disease. the recent advancement in crystallography and availability of crystal structures of certain domains of chikv initiates the development of anti-chikv agents using structure- ...201525921858
a neuron-specific antiviral mechanism prevents lethal flaviviral infection of mosquitoes.mosquitoes are natural vectors for many etiologic agents of human viral diseases. mosquito-borne flaviviruses can persistently infect the mosquito central nervous system without causing dramatic pathology or influencing the mosquito behavior and lifespan. the mechanism by which the mosquito nervous system resists flaviviral infection is still largely unknown. here we report that an aedes aegypti homologue of the neural factor hikaru genki (aahig) efficiently restricts flavivirus infection of the ...201525915054
modification of becn1 by isg15 plays a crucial role in autophagy regulation by type i ifn/interferon.isg15 (isg15 ubiquitin-like modifier), a ubiquitin-like protein, is one of the major type i ifn (interferon) effector systems. isg15 can be conjugated to target proteins (isgylation) via the stepwise action of e1, e2, and e3 enzymes. conjugated isg15 can be removed (deisgylated) from target proteins by usp18 (ubiquitin-specific peptidase 18). here we investigated the role of deisgylation by usp18 in regulating autophagy and egfr degradation in cells treated with type i ifns. we show that type i ...025906440
a small antigenic determinant of the chikungunya virus e2 protein is sufficient to induce neutralizing antibodies which are partially protective in mice.the mosquito-borne chikungunya virus (chikv) causes high fever and severe joint pain in humans. it is expected to spread in the future to europe and has recently reached the usa due to globalization, climate change and vector switch. despite this, little is known about the virus life cycle and, so far, there is no specific treatment or vaccination against chikungunya infections. we aimed here to identify small antigenic determinants of the chikv e2 protein able to induce neutralizing immune resp ...201525905779
culex torrentium mosquito role as major enzootic vector defined by rate of sindbis virus infection, sweden, 2009.we isolated sindbis virus (sinv) from the enzootic mosquito vectors culex torrentium, cx. pipiens, and culiseta morsitans collected in an area of sweden where sinv disease is endemic. the infection rate in cx. torrentium mosquitoes was exceptionally high (36 infections/1,000 mosquitoes), defining cx. torrentium as the main enzootic vector of sinv in scandinavia.025898013
[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
impacts of the creation, expansion and management of english wetlands on mosquito presence and abundance - developing strategies for future disease mitigation.the incidence of mosquito-borne diseases is increasing in europe, partly due to the incursion of a number of invasive species known to be vectors of dengue and chikungunya viruses, but also due to the involvement of native species in the transmission of west nile virus and malaria. for some of these pathogens, there is a risk of the re-emergence of vector-borne diseases that were once widespread in europe, but declined partly due to large-scale land-drainage projects. some mosquito species explo ...201525889666
understanding west nile virus ecology in europe: culex pipiens host feeding preference in a hotspot of virus emergence.understanding wildlife disease ecology is becoming an urgent need due to the continuous emergence and spread of several wildlife zoonotic diseases. west nile virus (wnv) is the most widespread arthropod-borne virus in the world, and in recent decades there has been an increase both in geographic range, and in the frequency of symptomatic infections in humans and wildlife. the principal vector for wnv in europe is the common house culex pipiens mosquito, which feeds on a wide variety of vertebrat ...201525888754
sequence analysis reveals a conserved extension in the capping enzyme of the alphavirus supergroup, and a homologous domain in nodaviruses.members of the alphavirus supergroup include human pathogens such as chikungunya virus, hepatitis e virus and rubella virus. they encode a capping enzyme with methyltransferase-guanylyltransferase (mtase-gtase) activity, which is an attractive drug target owing to its unique mechanism. however, its experimental study has proven very difficult.201525886938
endogenous sirnas and pirnas derived from transposable elements and genes in the malaria vector mosquito anopheles gambiae.the sirna and pirna pathways have been shown in insects to be essential for regulation of gene expression and defence against exogenous and endogenous genetic elements (viruses and transposable elements). the vast majority of endogenous small rnas produced by the sirna and pirna pathways originate from repetitive or transposable elements (te). in d. melanogaster, te-derived endogenous sirnas and pirnas are involved in genome surveillance and maintenance of genome integrity. in the medically rele ...201525879960
the brainstem oscillator for whisking and the case for breathing as the master clock for orofacial motor actions.whisking and sniffing are predominant aspects of exploratory behavior in rodents. we review evidence that these motor rhythms are coordinated by the respiratory patterning circuitry in the ventral medulla. a recently described region in the intermediate reticular zone of the medulla functions as an autonomous whisking oscillator, whose neuronal output is reset upon each breath by input from the pre-bötzinger complex. based on similarities between this neuronal circuit architecture and that of ot ...201525876629
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
structural basis of arc binding to synaptic proteins: implications for cognitive disease.arc is a cellular immediate-early gene (ieg) that functions at excitatory synapses and is required for learning and memory. we report crystal structures of arc subdomains that form a bi-lobar architecture remarkably similar to the capsid domain of human immunodeficiency virus (hiv) gag protein. analysis indicates arc originated from the ty3/gypsy retrotransposon family and was "domesticated" in higher vertebrates for synaptic functions. the arc n-terminal lobe evolved a unique hydrophobic pocket ...201525864631
generation of a safe and effective live viral vaccine by virus self-attenuation using species-specific artificial microrna.vaccination with live attenuated vaccines (lavs) is an effective way for prevention of infectious disease. while several methods are employed to create them, efficacy and safety are still a challenge. in this study, we evaluated the feasibility of creating a self-attenuated rna virus expressing a functional species-specific artificial microrna. using influenza virus as a model, we produced an attenuated virus carrying a mammalian-specific mir-93 expression cassette that expresses a viral nucleop ...201525858415
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
the transcription factor foxk participates with nup98 to regulate antiviral gene expression.upon infection, pathogen recognition leads to a rapidly activated gene expression program that induces antimicrobial effectors to clear the invader. we recently found that nup98 regulates the expression of a subset of rapidly activated antiviral genes to restrict disparate rna virus infections in drosophila by promoting rna polymerase occupancy at the promoters of these antiviral genes. how nup98 specifically targets these loci was unclear; however, it is known that nup98 participates with trans ...201525852164
noncapped alphavirus genomic rnas and their role during infection.alphaviruses are enveloped positive-sense rna viruses that exhibit a wide host range consisting of vertebrate and invertebrate species. previously we have reported that the infectivity of sindbis virus (sinv), the model alphavirus, was largely a function of the cell line producing the viral particles. mammalian-cell-derived sinv particles, on average, exhibit a higher particle-to-pfu ratio than mosquito cell-derived sinv particles. nevertheless, the outcome of nonproductive infection, the molecu ...201525833042
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
virus-induced dormancy in the archaeon sulfolobus islandicus.we investigated the interaction between sulfolobus spindle-shaped virus (ssv9) and its native archaeal host sulfolobus islandicus. we show that upon exposure to ssv9, s. islandicus strain rjw002 has a significant growth delay where the majority of cells are dormant (viable but not growing) for 24 to 48 hours postinfection (hpi) compared to the growth of controls without virus. we demonstrate that in this system, dormancy (i) is induced by both active and inactive virus particles at a low multipl ...201525827422
nuclear proteins hijacked by mammalian cytoplasmic plus strand rna strand rna viruses that replicate in the cytoplasm face challenges in supporting the numerous biosynthetic functions required for replication and propagation. most of these viruses are genetically simple and rely heavily on co-opting cellular proteins, particularly cellular rna-binding proteins, into new roles for support of virus infection at the level of virus-specific translation, and building rna replication complexes. in the course of infectious cycles many nuclear-cytoplasmic shuttlin ...201525818028
autophagy mediates tolerance to staphylococcus aureus alpha-toxin.resistance and tolerance are two defense strategies employed by the host against microbial threats. autophagy-mediated degradation of bacteria has been extensively described as a major resistance mechanism. here we find that the dominant function of autophagy proteins during infections with the epidemic community-associated methicillin-resistant staphylococcus aureus usa300 is to mediate tolerance rather than resistance. atg16l1 hypomorphic mice (atg16l1(hm)), which have reduced autophagy, were ...201525816775
parallel epigenomic and transcriptomic responses to viral infection in honey bees (apis mellifera).populations of honey bees are declining throughout the world, with us beekeepers losing 30% of their colonies each winter. though multiple factors are driving these colony losses, it is increasingly clear that viruses play a major role. however, information about the molecular mechanisms mediating antiviral immunity in honey bees is surprisingly limited. here, we examined the transcriptional and epigenetic (dna methylation) responses to viral infection in honey bee workers. one-day old worker ho ...201525811620
catch me if you can: the link between autophagy and viruses. 201525811485
dissection of amino-terminal functional domains of murine coronavirus nonstructural protein 3.coronaviruses, the largest rna viruses, have a complex program of rna synthesis that entails genome replication and transcription of subgenomic mrnas. rna synthesis by the prototype coronavirus mouse hepatitis virus (mhv) is carried out by a replicase-transcriptase composed of 16 nonstructural protein (nsp) subunits. among these, nsp3 is the largest and the first to be inserted into the endoplasmic reticulum. nsp3 comprises multiple structural domains, including two papain-like proteases (plps) ...201525810552
mammalian synthetic biology: emerging medical this review, we discuss new emerging medical applications of the rapidly evolving field of mammalian synthetic biology. we start with simple mammalian synthetic biological components and move towards more complex and therapy-oriented gene circuits. a comprehensive list of on-off switches, categorized into transcriptional, post-transcriptional, translational and post-translational, is presented in the first sections. subsequently, boolean logic gates, synthetic mammalian oscillators and toggle ...201525808341
viral rna intermediates as targets for detection and discovery of novel and emerging mosquito-borne viruses.mosquito-borne viruses encompass a range of virus families, comprising a number of significant human pathogens (e.g., dengue viruses, west nile virus, chikungunya virus). virulent strains of these viruses are continually evolving and expanding their geographic range, thus rapid and sensitive screening assays are required to detect emerging viruses and monitor their prevalence and spread in mosquito populations. double-stranded rna (dsrna) is produced during the replication of many of these virus ...201525799391
viral noncoding rnas: more surprises.eukaryotic cells produce several classes of long and small noncoding rna (ncrna). many dna and rna viruses synthesize their own ncrnas. like their host counterparts, viral ncrnas associate with proteins that are essential for their stability, function, or both. diverse biological roles--including the regulation of viral replication, viral persistence, host immune evasion, and cellular transformation--have been ascribed to viral ncrnas. in this review, we focus on the multitude of functions playe ...025792595
antiviral and immunoregulatory effects of indoleamine-2,3-dioxygenase in hepatitis c virus patients with hepatitis c virus (hcv) infection, enhanced activity of indoleamine-2,3-dioxygenase 1 (ido) has been reported. ido - a tryptophan-catabolizing enzyme - has been considered as both an innate defence mechanism and an important regulator of the immune response. the molecular mechanism of ido induction in hcv infection and its role in the antiviral immune response remain unknown. using primary human hepatocytes, we show that hcv infection stimulates ido expression. ido gene inductio ...201525792183
recent advances and the future of stem cell therapies in amyotrophic lateral sclerosis.amyotrophic lateral sclerosis is a progressive neurodegenerative disease of the motor neurons without a known cure. based on the possibility of cellular neuroprotection and early preclinical results, stem cells have gained widespread enthusiasm as a potential treatment strategy. preclinical models demonstrate a protective role of engrafted stem cells and provided the basis for human trials carried out using various types of stem cells, as well as a range of cell delivery methods. to date, no tri ...025776222
the hub protein loquacious connects the microrna and short interfering rna pathways in mosquitoes.aedes aegypti mosquitoes vector several arboviruses of global health significance, including dengue viruses and chikungunya virus. rna interference (rnai) plays an important role in antiviral immunity, gene regulation and protection from transposable elements. double-stranded rna binding proteins (dsrbps) are important for efficient rnai; in drosophila functional specialization of the mirna, endo-sirna and exo-sirna pathway is aided by the dsrbps loquacious (loqs-pb, loqs-pd) and r2d2, respectiv ...201525765650
mechanisms of innate immune evasion in re-emerging rna viruses.recent outbreaks of ebola, west nile, chikungunya, middle eastern respiratory and other emerging/re-emerging rna viruses continue to highlight the need to further understand the virus-host interactions that govern disease severity and infection outcome. as part of the early host antiviral defense, the innate immune system mediates pathogen recognition and initiation of potent antiviral programs that serve to limit virus replication, limit virus spread and activate adaptive immune responses. conc ...201525765605
de novo assembly and transcriptome analysis of atlantic salmon macrophage/dendritic-like to cells following type i ifn treatment and salmonid alphavirus subtype-3 infection.interferons (ifn) are cytokines secreted by vertebrate cells involved in activation of signaling pathways that direct the synthesis of antiviral genes. to gain a global understanding of antiviral genes induced by type i ifns in salmonids, we used rna-seq to characterize the transcriptomic changes induced by type i ifn treatment and salmon alphavirus subtype 3 (sav-3) infection in to-cells, a macrophage/dendritic like cell-line derived from atlantic salmon (salmo salar l) head kidney leukocytes.201525765343
sars hcov papain-like protease is a unique lys48 linkage-specific di-distributive deubiquitinating enzyme.ubiquitin (ub) and the ub-like (ubl) modifier interferon-stimulated gene 15 (isg15) participate in the host defence of viral infections. viruses, including the severe acute respiratory syndrome human coronavirus (sars hcov), have co-opted ub-isg15 conjugation pathways for their own advantage or have evolved effector proteins to counter pro-inflammatory properties of ub-isg15-conjugated host proteins. in the present study, we compare substrate specificities of the papain-like protease (plpro) fro ...025764917
activation of tomato bushy stunt virus rna-dependent rna polymerase by cellular heat shock protein 70 is enhanced by phospholipids in vitro.similar to other positive-strand rna viruses, tombusviruses are replicated by the membrane-bound viral replicase complex (vrc). the vrc consists of the p92 virus-coded rna-dependent rna polymerase (rdrp), the viral p33 rna chaperone, and several co-opted host proteins. in order to become a functional rdrp after its translation, the p92 replication protein should be incorporated into the vrc, followed by its activation. we have previously shown in a cell-free yeast extract-based assay that the ac ...201525762742
simultaneous recording of fluorescence and electrical signals by photometric patch electrode in deep brain regions in vivo.despite its widespread use, high-resolution imaging with multiphoton microscopy to record neuronal signals in vivo is limited to the surface of brain tissue because of limited light penetration. moreover, most imaging studies do not simultaneously record electrical neural activity, which is, however, crucial to understanding brain function. accordingly, we developed a photometric patch electrode (pme) to overcome the depth limitation of optical measurements and also enable the simultaneous recor ...201525761950
role of zinc-finger anti-viral protein in host defense against sindbis virus.accumulating evidence indicates that type i interferon (ifn) mediates the host protective response to rna viruses. however, the anti-viral effector molecules involved in this response have not been fully identified. here, we show that zinc-finger anti-viral protein (zap), an ifn-inducible gene, plays a critical role in the elimination of sindbis virus (sinv) in vitro and in vivo. the loss of zap greatly enhances the replication of sinv but does not inhibit type i ifn production in primary mouse ...201525758257
antiviral antibody profiling by high-density protein arrays.viral infections elicit antiviral antibodies and have been associated with various chronic diseases. detection of these antibodies can facilitate diagnosis, treatment of infection, and understanding of the mechanisms of virus-associated diseases. in this work, we assayed antiviral antibodies using a novel high-density nucleic acid programmable protein array (hd-nappa) platform. individual viral proteins were expressed in situ directly from plasmids encoding proteins in an array of microscopic re ...201525758251
hepatitis e: molecular virology and pathogenesis.hepatitis e virus is a single, positive-sense, capped and poly a tailed rna virus classified under the family hepeviridae. enteric transmission, acute self-limiting hepatitis, frequent epidemic and sporadic occurrence, high mortality in affected pregnants are hallmarks of hepatitis e infection. lack of an efficient culture system and resulting reductionist approaches for the study of replication and pathogenesis of hev made it to be a less understood agent. early studies on animal models, sub-ge ...201325755485
ltp-triggered cholesterol redistribution activates cdc42 and drives ampa receptor synaptic delivery.neurotransmitter receptor trafficking during synaptic plasticity requires the concerted action of multiple signaling pathways and the protein transport machinery. however, little is known about the contribution of lipid metabolism during these processes. in this paper, we addressed the question of the role of cholesterol in synaptic changes during long-term potentiation (ltp). we found that n-methyl-d-aspartate-type glutamate receptor (nmdar) activation during ltp induction leads to a rapid and ...201525753037
to translate, or not to translate: viral and host mrna regulation by interferon-stimulated genes.type i interferon (ifn) is one of the first lines of cellular defense against viral pathogens. as a result of ifn signaling, a wide array of ifn-stimulated gene (isg) products is upregulated to target different stages of the viral life cycle. we review recent findings implicating a subset of isgs in translational regulation of viral and host mrnas. translation inhibition is mediated either by binding to viral rna or by disrupting physiological interactions or levels of the translation complex co ...201525748385
xrn1 stalling in the 5' utr of hepatitis c virus and bovine viral diarrhea virus is associated with dysregulated host mrna stability.we demonstrate that both hepatitis c virus (hcv) and bovine viral diarrhea virus (bvdv) contain regions in their 5' utrs that stall and repress the enzymatic activity of the cellular 5'-3' exoribonuclease xrn1, resulting in dramatic changes in the stability of cellular mrnas. we used biochemical assays, virus infections, and transfection of the hcv and bvdv 5' untranslated regions in the absence of other viral gene products to directly demonstrate the existence and mechanism of this novel host-v ...201525747802
the role of misshapen nck-related kinase (mink), a novel ste20 family kinase, in the ires-mediated protein translation of human enterovirus 71.human enterovirus 71 (ev71) commonly causes hand, foot and mouth disease in young children, and occasional occurrences of neurological complications can be fatal. in this study, a high-throughput cell-based screening on the serine/threonine kinase sirna library was performed to identify potential antiviral agents against ev71 replication. among the hits, misshapen/niks-related kinase (mink) was selected for detailed analysis due to its strong inhibitory profile and novelty. in the investigation ...201525747578
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
description and utilization of the united states department of defense serum repository: a review of published studies, 1985-2012.specimens in the united states department of defense (dod) serum repository have accumulated in frozen storage since 1985 when the dod began universal screening for human immunodeficiency virus. use of the stored serum for health research has been carefully controlled, but the resulting publications have never been systematically identified or described. the armed forces health surveillance center (afhsc) information systems and open (online) sites were used as data sources. through 2012, the re ...201525723497
emerging roles for rna degradation in viral replication and antiviral defense.viral replication significantly alters the gene expression landscape of infected cells. many of these changes are driven by viral manipulation of host transcription or translation machinery. several mammalian viruses encode factors that broadly dampen gene expression by directly targeting messenger rna (mrna). here, we highlight how these factors promote mrna degradation to globally regulate both host and viral gene expression. although these viral factors are not homologous and use distinct mec ...201525721579
autophagy plays an important role in protecting pacific oysters from oshv-1 and vibrio aestuarianus infections.recent mass mortality outbreaks around the world in pacific oysters, crassostrea gigas, have seriously affected the aquaculture economy. although the causes for these mortality outbreaks appear complex, infectious agents are involved. two pathogens are associated with mass mortality outbreaks, the virus ostreid herpesvirus 1 (oshv-1) and the bacterium vibrio aestuarianus. here we describe the interactions between these 2 pathogens and autophagy, a conserved intracellular pathway playing a key ro ...025714877
binding of the pathogen receptor hsp90aa1 to avibirnavirus vp2 induces autophagy by inactivating the akt-mtor pathway.autophagy is an essential component of host innate and adaptive immunity. viruses have developed diverse strategies for evading or utilizing autophagy for survival. the response of the autophagy pathways to virus invasion is poorly documented. here, we report on the induction of autophagy initiated by the pathogen receptor hsp90aa1 (heat shock protein 90 kda α [cytosolic], class a member 1) via the akt-mtor (mechanistic target of rapamycin)-dependent pathway. transmission electron microscopy and ...025714412
rapid selection against arbovirus-induced apoptosis during infection of a mosquito vector.millions of people are infected each year by arboviruses (arthropod-borne viruses) such as chikungunya, dengue, and west nile viruses, yet for reasons that are largely unknown, only a relatively small number of mosquito species are able to transmit arboviruses. understanding the complex factors that determine vector competence could facilitate strategies for controlling arbovirus infections. apoptosis is a potential antiviral defense response that has been shown to be important in other virus-ho ...201525713358
rna degradation in antiviral immunity and control determines the fate of cellular rna molecules. nonsense-mediated decay (nmd) provides quality control of mrna, targeting faulty cellular transcripts for degradation by multiple nucleases including the rna exosome. recent findings have revealed a role for nmd in targeting viral rna molecules, thereby restricting virus infection. interestingly, nmd is also linked to immune responses at another level: mutations affecting the nmd or rna exosome machineries cause chronic ...201525709093
the α2,3-sialyltransferase encoded by myxoma virus is a virulence factor that contributes to immunosuppression.myxoma virus (myxv) induces a lethal disease called myxomatosis in european rabbits. myxv is one of the rare viruses that encodes an α2,3-sialyltransferase through its m138l gene. in this study, we showed that although the absence of the enzyme was not associated with any in vitro deficit, the m138l deficient strains are highly attenuated in vivo. indeed, while all rabbits infected with the parental and the revertant strains died within 9 days post-infection from severe myxomatosis, all but one ...201525705900
new tricks for old dogs: countering antibiotic resistance in tuberculosis with host-directed therapeutics.despite the availability of mycobacterium tuberculosis (mtb) drugs for over 50 years, tuberculosis (tb) remains at pandemic levels. new drugs are urgently needed for resistant strains, shortening duration of treatment, and targeting different stages of the disease, especially for treatment during human immunodeficiency virus co-infection. one solution to the conundrum that antibiotics kill the bacillus yet select for resistance is to target the host rather than the pathogen. here, we discuss rec ...025703571
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
seroepidemiology of selected arboviruses in febrile patients visiting selected health facilities in the lake/river basin areas of lake baringo, lake naivasha, and tana river, kenya.arboviruses cause emerging and re-emerging infections affecting humans and animals. they are spread primarily by blood-sucking insects such as mosquitoes, ticks, midges, and sandflies. changes in climate, ecology, demographic, land-use patterns, and increasing global travel have been linked to an upsurge in arboviral disease. outbreaks occur periodically followed by persistent low-level circulation.025700043
functional analysis of the short isoform of orf virus protein ov20.0.orf virus (orfv) ov20.0l is an ortholog of vaccinia virus (vacv) gene e3l. the function of vacv e3 protein as a virulence factor is well studied, but ov20.0 has received less attention. here we show that like vacv e3l, ov20.0l encodes two proteins, a full-length protein and a shorter form (sh20). the shorter sh20 is an n-terminally truncated ov20.0 isoform generated when a downstream aug codon is used for initiating translation. these isoforms differed in cellular localization, with full-length ...201525694596
a basic cluster in the n terminus of yellow fever virus ns2a contributes to infectious particle production.the flavivirus ns2a protein is involved in the assembly of infectious particles. to further understand its role in this process, a charged-to-alanine scanning analysis was performed on ns2a encoded by an infectious cdna clone of yellow fever virus (yfv). fifteen mutants containing single, double, or triple charged-to-alanine changes were tested. five of them did not produce infectious particles, whereas efficient rna replication was detectable for two of the five ns2a mutants (r22a-k23a-r24a and ...201525694595
the laminin receptor is a cellular attachment receptor for classical swine fever virus.classical swine fever virus (csfv) is the causative agent of classical swine fever (csf), a highly contagious, economically important viral disease in many countries. the e(rns) and e2 envelope glycoproteins are responsible for the binding to and entry into the host cell by csfv. to date, only one cellular receptor, heparan sulfate (hs), has been identified as being involved in csfv attachment. hs is also present on the surface of various cells that are nonpermissive to csfv. hence, there must b ...201525694590
dendritic geometry shapes neuronal camp signalling to the nucleus.neurons have complex dendritic trees, receiving numerous inputs at various distances from the cell body. yet the rules of molecular signal propagation from dendrites to nuclei are unknown. darpp-32 is a phosphorylation-regulated signalling hub in striatal output neurons. we combine diffusion-reaction modelling and live imaging to investigate camp-activated darpp-32 signalling to the nucleus. the model predicts maximal effects on the nucleus of camp production in secondary dendrites, due to segme ...201525692798
discovery of itraconazole with broad-spectrum in vitro antienterovirus activity that targets nonstructural protein 3a.there is currently no approved antiviral therapy for the prophylaxis or treatment of enterovirus infections, which remain a substantial threat to public health. to discover inhibitors that can be immediately repurposed for treatment of enterovirus infections, we developed a high-throughput screening assay that measures the cytopathic effect induced by enterovirus 71 (ev71) to screen an fda-approved drug library. itraconazole (itz), a triazole antifungal agent, was identified as an effective inhi ...201525691649
the role of rna interference (rnai) in arbovirus-vector interactions.rna interference (rnai) was shown over 18 years ago to be a mechanism by which arbovirus replication and transmission could be controlled in arthropod vectors. during the intervening period, research on rnai has defined many of the components and mechanisms of this antiviral pathway in arthropods, yet a number of unexplored questions remain. rnai refers to rna-mediated regulation of gene expression. originally, the term described silencing of endogenous genes by introduction of exogenous double- ...201525690800
climate, environmental and socio-economic change: weighing up the balance in vector-borne disease transmission.arguably one of the most important effects of climate change is the potential impact on human health. while this is likely to take many forms, the implications for future transmission of vector-borne diseases (vbds), given their ongoing contribution to global disease burden, are both extremely important and highly uncertain. in part, this is owing not only to data limitations and methodological challenges when integrating climate-driven vbd models and climate change projections, but also, perhap ...025688012
shp-1-dependent macrophage differentiation exacerbates virus-induced myositis.virus-induced myositis is an emerging global affliction that remains poorly characterized with few treatment options. moreover, muscle-tropic viruses often spread to the cns, causing dramatically increased morbidity. therefore, there is an urgent need to explore genetic factors involved in this class of human disease. this report investigates critical innate immune pathways affecting murine virus-induced myositis. of particular importance, the key immune regulator src homology region 2 domain-co ...201525681345
putative novel lineage of west nile virus in uranotaenia unguiculata mosquito, hungary.west nile virus (wnv) is an increasing public health concern in europe with numerous human cases. a total of 23,029 female mosquitoes were tested for a variety of mosquito-borne flaviviruses and orthobunyaviruses supposedly endemic in southern transdanubia, hungary, in the frames of a large-scale surveillance between 2011 and 2013. wnv nucleic acid was detected in a single pool containing uranotaenia unguiculata mosquitoes. sequence- and phylogenetic analyses for two different regions (ns5 and e ...201425674630
initial evidence on differences among enterovirus 71, coxsackievirus a16 and coxsackievirus b4 in binding to cell surface heparan sulphate.cell surface heparan sulphate (hs) mediates infection for many viruses from diverse families. we demonstrated significant antiviral potencies for a number of hs mimetics against a cloned strain of enterovirus 71 (ev71) in a previous study. thus, the involvement of hs in mediating viral infection of isolates of human enteroviruses was investigated in vero and human neural cells in the present work. in both cell lines, heparin and pentosan polysulphate significantly inhibited both infection and at ...201325674594
antiviral responses of arthropod vectors: an update on recent advances.arthropod vectors, such as mosquitoes, ticks, biting midges and sand flies, transmit many viruses that can cause outbreaks of disease in humans and animals around the world. arthropod vector species are invading new areas due to globalisation and environmental changes, and contact between exotic animal species, humans and arthropod vectors is increasing, bringing with it the regular emergence of new arboviruses. for future strategies to control arbovirus transmission, it is important to improve ...201425674592
virological and preclinical characterization of a dendritic cell targeting, integration-deficient lentiviral vector for cancer immunotherapy.dendritic cells (dcs) are essential antigen-presenting cells for the initiation of cytotoxic t-cell responses and therefore attractive targets for cancer immunotherapy. we have developed an integration-deficient lentiviral vector termed id-vp02 that is designed to deliver antigen-encoding nucleic acids selectively to human dcs in vivo. id-vp02 utilizes a genetically and glycobiologically engineered sindbis virus glycoprotein to target human dcs through the c-type lectin dc-sign (cd209) and also ...025658613
tnbp⁄triton x-45 treatment of plasma for transfusion efficiently inactivates hepatitis c virus.risk of transmission of hepatitis c virus (hcv) by clinical plasma remains high in countries with a high prevalence of hepatitis c, justifying the implementation of viral inactivation treatments. in this study, we assessed the extent of inactivation of hcv during minipool solvent/detergent (sd; 1% tnbp / 1% triton x-45) treatment of human plasma. luciferase-tagged infectious cell culture-derived hcv (hcvcc) particles were used to spike human plasma prior to treatment by sd at 31 ± 0.5°c for 30 m ...201525658612
viral and cellular proteins containing fgdf motifs bind g3bp to block stress granule formation.the ras-gap sh3 domain-binding proteins (g3bp) are essential regulators of the formation of stress granules (sg), cytosolic aggregates of proteins and rna that are induced upon cellular stress, such as virus infection. many viruses, including semliki forest virus (sfv), block sg induction by targeting g3bp. in this work, we demonstrate that the g3bp-binding motif of sfv nsp3 consists of two fgdf motifs, in which both phenylalanine and the glycine residue are essential for binding. in addition, w ...201525658430
stress granule components g3bp1 and g3bp2 play a proviral role early in chikungunya virus replication.stress granules (sgs) are protein-mrna aggregates that are formed in response to environmental stresses, resulting in translational inhibition. sgs are generally believed to play an antiviral role and are manipulated by many viruses, including various alphaviruses. gtpase-activating protein (sh3 domain)-binding protein 1 (g3bp1) is a key component and commonly used marker of sgs. its homolog g3bp2 is a less extensively studied sg component. here, we demonstrate that chikungunya virus (chikv) inf ...201525653451
atp1a1-mediated src signaling inhibits coronavirus entry into host addition to transporting ions, the multisubunit na(+),k(+)-atpase also functions by relaying cardiotonic steroid (cts)-binding-induced signals into cells. in this study, we analyzed the role of na(+),k(+)-atpase and, in particular, of its atp1a1 α subunit during coronavirus (cov) infection. as controls, the vesicular stomatitis virus (vsv) and influenza a virus (iav) were included. using gene silencing, the atp1a1 protein was shown to be critical for infection of cells with murine hepatitis v ...201525653449
residue-level resolution of alphavirus envelope protein interactions in ph-dependent fusion.alphavirus envelope proteins, organized as trimers of e2-e1 heterodimers on the surface of the pathogenic alphavirus, mediate the low ph-triggered fusion of viral and endosomal membranes in human cells. the lack of specific treatment for alphaviral infections motivates our exploration of potential antiviral approaches by inhibiting one or more fusion steps in the common endocytic viral entry pathway. in this work, we performed constant ph molecular dynamics based on an atomic model of the alphav ...201525646410
neurological sequelae induced by alphavirus infection of the cns are attenuated by treatment with the glutamine antagonist 6-diazo-5-oxo-l-norleucine.recovery from encephalomyelitis induced by infection with mosquito-borne alphaviruses is associated with a high risk of lifelong debilitating neurological deficits. infection of mice with the prototypic alphavirus, sindbis virus, provides an animal model with which to study disease mechanisms and examine potential therapeutics. infectious virus is cleared from the brain within a week after infection, but viral rna is cleared slowly and persists for the life of the animal. however, no studies hav ...201525645378
bovine lactoferrin inhibits toscana virus infection by binding to heparan sulphate.toscana virus is an emerging sandfly-borne bunyavirus in mediterranean europe responsible for neurological diseases in humans. it accounts for about 80% of paediatric meningitis cases during the summer. despite the important impact of toscana virus infection-associated disease on human health, currently approved vaccines or effective antiviral treatments are not available. in this research, we have analyzed the effect of bovine lactoferrin, a bi-globular iron-binding glycoprotein with potent ant ...201525643293
sindbis virus interferes with dengue 4 virus replication and its potential transmission by aedes albopictus.mosquitoes transmit a number of arboviruses associated with disease outbreaks in humans and other animals. the majority of medically important arboviruses belong to three families: togaviridae, flaviviridae and bunyaviridae. several members of these families have overlapping distributions and share common vectors, increasing the potential for arboviral coinfections. this study examined how two model viruses: sindbis virus (sinv, togaviridae: alphavirus) and dengue-4 virus (denv-4, flaviviridae: ...201525633862
the 5' and 3' ends of alphavirus rnas--non-coding is not non-functional.the non-coding regions found at the 5' and 3' ends of alphavirus genomes regulate viral gene expression, replication, translation and virus-host interactions, which have significant implications for viral evolution, host range, and pathogenesis. the functions of these non-coding regions are mediated by a combination of linear sequence and structural elements. the capped 5' untranslated region (utr) contains promoter elements, translational regulatory sequences that modulate dependence on cellula ...201525630058
teaching the basics of autophagy and mitophagy to redox biologists--mechanisms and experimental approaches.autophagy is a lysosomal mediated degradation activity providing an essential mechanism for recycling cellular constituents, and clearance of excess or damaged lipids, proteins and organelles. autophagy involves more than 30 proteins and is regulated by nutrient availability, and various stress sensing signaling pathways. this article provides an overview of the mechanisms and regulation of autophagy, its role in health and diseases, and methods for its measurement. hopefully this teaching revie ...201525618581
pathogen-inspired drug delivery to the central nervous system.for as long as the human blood-brain barrier (bbb) has been evolving to exclude bloodborne agents from the central nervous system (cns), pathogens have adopted a multitude of strategies to bypass it. some pathogens, notably viruses and certain bacteria, enter the cns in whole form, achieving direct physical passage through endothelial or neuronal cells to infect the brain. other pathogens, including bacteria and multicellular eukaryotic organisms, secrete toxins that preferentially interact with ...201425610755
a polyprotein-expressing salmonid alphavirus replicon induces modest protection in atlantic salmon (salmo salar) against infectious pancreatic necrosis.vaccination is an important strategy for the control and prevention of infectious pancreatic necrosis (ipn) in farmed atlantic salmon (salmo salar) in the post-smolt stage in sea-water. in this study, a heterologous gene expression system, based on a replicon construct of salmonid alphavirus (sav), was used for in vitro and in vivo expression of ipn virus proteins. the large open reading frame of segment a, encoding the polyprotein nh2-pvp2-vp4-vp3-cooh, as well as pvp2, were cloned and expresse ...201525606973
in- silico exploration of thirty alphavirus genomes for analysis of the simple sequence repeats.the compilation of simple sequence repeats (ssrs) in viruses and its analysis with reference to incidence, distribution and variation would be instrumental in understanding the functional and evolutionary aspects of repeat sequences. present study encompasses the analysis of ssrs across 30 species of alphaviruses. the full length genome sequences, assessed from ncbi were used for extraction and analysis of repeat sequences using imex software. the repeats of different motif sizes (mono- to penta ...201425606453
selective inactivation of usp18 isopeptidase activity in vivo enhances isg15 conjugation and viral resistance.protein modification by the ubiquitin-like protein isg15 is an interferon (ifn) effector system, which plays a major role in antiviral defense. isg15 modification is counteracted by the isopeptidase usp18, a major negative regulator of ifn signaling, which was also shown to exert its regulatory function in an isopeptidase-independent manner. to dissect enzymatic and nonenzymatic functions of usp18 in vivo, we generated knock-in mice (usp18(c61a/c61a)) expressing enzymatically inactive usp18. usp ...201525605921
adp-ribosylation factor 6 (arf6) bidirectionally regulates dendritic spine formation depending on neuronal maturation and activity.recent studies have reported conflicting results regarding the role of arf6 in dendritic spine development, but no clear answer for the controversy has been suggested. we found that adp-ribosylation factor 6 (arf6) either positively or negatively regulates dendritic spine formation depending on neuronal maturation and activity. arf6 activation increased the spine formation in developing neurons, whereas it decreased spine density in mature neurons. genome-wide microarray analysis revealed that a ...201525605715
ifitm-family proteins: the cell's first line of antiviral defense.animal cells use a wide variety of mechanisms to slow or prevent replication of viruses. these mechanisms are usually mediated by antiviral proteins whose expression and activities can be constitutive but are frequently amplified by interferon induction. among these interferon-stimulated proteins, members of the ifitm (interferon-induced transmembrane) family are unique because they prevent infection before a virus can traverse the lipid bilayer of the cell. at least three human ifitm proteins-i ...025599080
in vitro and in vivo regulation of synaptogenesis by the novel antidepressant spadin.we have described a novel antidepressant peptide, spadin, that acts by blocking the twik-related-potassium channel, type 1 (trek-1). here, we examined possible mechanisms of action of spadin at both molecular and cellular levels.201525598009
virus-host interactomics: new insights and opportunities for antiviral drug discovery.the current therapeutic arsenal against viral infections remains limited, with often poor efficacy and incomplete coverage, and appears inadequate to face the emergence of drug resistance. our understanding of viral biology and pathophysiology and our ability to develop a more effective antiviral arsenal would greatly benefit from a more comprehensive picture of the events that lead to viral replication and associated symptoms. towards this goal, the construction of virus-host interactomes is in ...201425593595
relationships among dissemination of primary parainfluenza virus infection in the respiratory tract, mucosal and peripheral immune responses, and protection from reinfection: a noninvasive bioluminescence-imaging study.respiratory paramyxoviruses such as respiratory syncytial virus (rsv) and human parainfluenza virus type 1 (hpiv1) to hpiv4 infect virtually all children by the age of 2 to 5 years, leading to partial but incomplete protection from reinfection. here, we used luciferase-expressing reporter sendai viruses (the murine counterpart of hpiv1) to noninvasively measure primary infection, immune responses, and protection from reinfection by either a lethal challenge or natural transmission in living mice ...201525589649
orientation refinement of virus structures with unknown symmetry.structural biology, in particular the structure determination of viruses and other large macromolecular complexes leads to data- and compute-intensive problems that require resources well beyond those available on a single system. thus, there is an imperative need to develop parallel algorithms and programs for clusters and computational grids. we present one of the most challenging computational problems posed by the three-dimensional structure determination of viruses, the orientation refineme ...025580470
identification of hnrnpk as regulator of hepatitis c virus particle production.hepatitis c virus (hcv) is a major cause of chronic liver disease affecting around 130 million people worldwide. while great progress has been made to define the principle steps of the viral life cycle, detailed knowledge how hcv interacts with its host cells is still limited. to overcome this limitation we conducted a comprehensive whole-virus rna interference-based screen and identified 40 host dependency and 16 host restriction factors involved in hcv entry/replication or assembly/release. of ...201525569684
early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification.a method for the rapid diagnosis of early dengue virus (denv) infection is highly needed. here, a prototype reverse transcription-recombinase polymerase amplification (rt-rpa) assay was developed. the assay detected denv rna in <20 min without the need for thermocycling amplification. the assay enabled the detection of as few as 10 copies of denv rna. the designed rt-rpa primers and exo probe detected the denv genome of at least 12 genotypes of denv circulating globally without cross-reacting wi ...201525568438
conserved motifs in a tombusvirus polymerase modulate genome replication, subgenomic transcription, and amplification of defective interfering rnas.the replication of plus-strand rna virus genomes is mediated by virally encoded rna-dependent rna polymerases (rdrps). we have investigated the role of the c-proximal region in the rdrp of tomato bushy stunt virus (tbsv) in mediating viral rna synthesis. tbsv is the prototype species in the genus tombusvirus, family tombusviridae, and its rdrp is responsible for replicating the viral genome, transcribing two subgenomic mrnas, and supporting replication of defective interfering rnas. comparative ...201525568204
vpx-containing dendritic cell vaccine induces ctls and reactivates latent hiv-1 in vitro.eradication of human immunodeficiency virus-1 (hiv-1) from an infected individual requires a means of inducing production of virus from latently infected cells and stimulating an immune response against the infected cells. we report the development of lentiviral vectors that transduce dendritic cells (dcs) to both induce production of virus from latently infected cells and stimulate antigen-specific cytotoxic t lymphocytes (ctls). the vectors package vpx, a lentiviral accessory protein that coun ...201525567537
Displaying items 401 - 500 of 5405