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pathogenic chikungunya virus evades b cell responses to establish persistence.chikungunya virus (chikv) and related alphaviruses cause epidemics of acute and chronic musculoskeletal disease. to investigate the mechanisms underlying the failure of immune clearance of chikv, we studied mice infected with an attenuated chikv strain (181/25) and the pathogenic parental strain (af15561), which differ by five amino acids. whereas af15561 infection of wild-type mice results in viral persistence in joint tissues, 181/25 is cleared. in contrast, 181/25 infection of μmt mice lackin ...201627452455
n-terminal domain of feline calicivirus (fcv) proteinase-polymerase contributes to the inhibition of host cell transcription.feline calicivirus (fcv) infection results in the inhibition of host protein synthesis, known as "shut-off". however, the precise mechanism of shut-off remains unknown. here, we found that the fcv strain 2280 proteinase-polymerase (pp) protein can suppress luciferase reporter gene expression driven by endogenous and exogenous promoters. furthermore, we found that the n-terminal 263 aa of pp (ppn-263) determined its shut-off activity using the expression of truncated proteins. however, the same d ...201627447663
recent advances in lentiviral vaccines for hiv-1 infection.the development of an effective hiv vaccine to prevent and/or cure hiv remains a global health priority. given their central role in the initiation of adaptive immune responses, dendritic cell (dc)-based vaccines are being increasingly explored as immunotherapeutic strategies to enhance hiv-specific t cells in infected individuals and, thus, promote immune responses that may help facilitate a functional cure. hiv-1-based lentiviral (lv) vectors have inherent advantages as dc vaccine vectors due ...201627446074
exaptive origins of regulated mrna decay in eukaryotes.eukaryotic gene expression is extensively controlled at the level of mrna stability and the mechanisms underlying this regulation are markedly different from their archaeal and bacterial counterparts. we propose that two such mechanisms, nonsense-mediated decay (nmd) and motif-specific transcript destabilization by ccch-type zinc finger rna-binding proteins, originated as a part of cellular defense against rna pathogens. these branches of the mrna turnover pathway might have been used by primeva ...201627438915
ribosomal frameshifting and transcriptional slippage: from genetic steganography and cryptography to adventitious use.genetic decoding is not 'frozen' as was earlier thought, but dynamic. one facet of this is frameshifting that often results in synthesis of a c-terminal region encoded by a new frame. ribosomal frameshifting is utilized for the synthesis of additional products, for regulatory purposes and for translational 'correction' of problem or 'savior' indels. utilization for synthesis of additional products occurs prominently in the decoding of mobile chromosomal element and viral genomes. one class of re ...201627436286
single-site glycoprotein mutants inhibit a late event in sindbis virus assembly.a panel of sindbis virus mutants that were suspected to have deficiencies in one or more aspects of their replication cycles was examined in baby hamster kidney (bhk) cells. these included an amino acid deletion (δh230) and substitution (h230a) in the sindbis glycoprotein e1_h230 and similar mutants in e2_g209 (g209a, g209d, and δg209). neither h230 mutation produced a measurable titer, but repeated passaging of the h230a mutant in bhk cells produced a second-site compensatory mutant (v231i) tha ...201627412592
larval food quantity affects the capacity of adult mosquitoes to transmit human malaria.adult traits of holometabolous insects are shaped by conditions experienced during larval development, which might impact interactions between adult insect hosts and parasites. however, the ecology of larval insects that vector disease remains poorly understood. here, we used anopheles stephensi mosquitoes and the human malaria parasite plasmodium falciparum, to investigate whether larval conditions affect the capacity of adult mosquitoes to transmit malaria. we reared larvae in two groups; one ...027412284
health technology assessment of pathogen reduction technologies applied to plasma for clinical use.although existing clinical evidence shows that the transfusion of blood components is becoming increasingly safe, the risk of transmission of known and unknown pathogens, new pathogens or re-emerging pathogens still persists. pathogen reduction technologies may offer a new approach to increase blood safety. the study is the output of collaboration between the italian national blood centre and the post-graduate school of health economics and management, catholic university of the sacred heart, ro ...201627403740
mechanisms of viral mutation.the remarkable capacity of some viruses to adapt to new hosts and environments is highly dependent on their ability to generate de novo diversity in a short period of time. rates of spontaneous mutation vary amply among viruses. rna viruses mutate faster than dna viruses, single-stranded viruses mutate faster than double-strand virus, and genome size appears to correlate negatively with mutation rate. viral mutation rates are modulated at different levels, including polymerase fidelity, sequence ...201627392606
alphavirus protease inhibitors from natural sources: a homology modeling and molecular docking investigation.alphaviruses such as chikungunya virus (chikv), o'nyong-nyong virus (onnv), ross river virus (rrv), eastern equine encephalitis virus (eeev), venezuelan equine encephalitis virus (veev), and western equine encephalitis virus (weev), are mosquito-transmitted viruses that can cause fevers, rash, and rheumatic diseases (chikv, onnv, rrv) or potentially fatal encephalitis (eeev, veev, weev) in humans. these diseases are considered neglected tropical diseases for which there are no current antiviral ...201627387412
genetic dissection of flaviviridae host factors through genome-scale crispr screens.the flaviviridae are a family of viruses that cause severe human diseases. for example, dengue virus (denv) is a rapidly emerging pathogen causing an estimated 100 million symptomatic infections annually worldwide. no approved antivirals are available to date and clinical trials with a tetravalent dengue vaccine showed disappointingly low protection rates. hepatitis c virus (hcv) also remains a major medical problem, with 160 million chronically infected patients worldwide and only expensive tre ...027383987
combined structural, biochemical and cellular evidence demonstrates that both fgdf motifs in alphavirus nsp3 are required for efficient replication.recent findings have highlighted the role of the old world alphavirus non-structural protein 3 (nsp3) as a host defence modulator that functions by disrupting stress granules, subcellular phase-dense rna/protein structures formed upon environmental stress. this disruption mechanism was largely explained through nsp3-mediated recruitment of the host g3bp protein via two tandem fgdf motifs. here, we present the 1.9 å resolution crystal structure of the ntf2-like domain of g3bp-1 in complex with a ...027383630
neurological sequelae resulting from encephalitic alphavirus infection.the recent surge in viral clinical cases and associated neurological deficits have reminded us that viral infections can lead to detrimental, long-term effects, termed sequelae, in survivors. alphaviruses are enveloped, single-stranded positive-sense rna viruses in the togaviridae family. transmission of alphaviruses between and within species occurs mainly via the bite of an infected mosquito bite, giving alphaviruses a place among arboviruses, or arthropod-borne viruses. alphaviruses are found ...201627379085
viral inhibition of the ifn-induced jak/stat signalling pathway: development of live attenuated vaccines by mutation of viral-encoded ifn-antagonists.the interferon (ifn) induced anti-viral response is amongst the earliest and most potent of the innate responses to fight viral infection. the induction of the janus kinase/signal transducer and activation of transcription (jak/stat) signalling pathway by ifns leads to the upregulation of hundreds of interferon stimulated genes (isgs) for which, many have the ability to rapidly kill viruses within infected cells. during the long course of evolution, viruses have evolved an extraordinary range of ...201627367734
who regulates whom? an overview of rna granules and viral infections.after viral infection, host cells respond by mounting an anti-viral stress response in order to create a hostile atmosphere for viral replication, leading to the shut-off of mrna translation (protein synthesis) and the assembly of rna granules. two of these rna granules have been well characterized in yeast and mammalian cells, stress granules (sgs), which are translationally silent sites of rna triage and processing bodies (pbs), which are involved in mrna degradation. this review discusses the ...201627367717
dissecting virus infectious cycles by cryo-electron microscopy. 201627362353
pirna pathway is not required for antiviral defense in drosophila melanogaster.since its discovery, rna interference has been identified as involved in many different cellular processes, and as a natural antiviral response in plants, nematodes, and insects. in insects, the small interfering rna (sirna) pathway is the major antiviral response. in recent years, the piwi-interacting rna (pirna) pathway also has been implicated in antiviral defense in mosquitoes infected with arboviruses. using drosophila melanogaster and an array of viruses that infect the fruit fly acutely o ...201627357659
the strica homolog aacasps16 is involved in apoptosis in the yellow fever vector, aedes albopictus.caspases are a family of cysteine proteases playing essential roles during apoptosis. seven caspases identified in drosophila were dronc, dredd, strica, dcp-1, decay, drice and damm. among them, strica is an insect-specific caspase containing a long serine- and threonine- rich prodomain, of which function is not yet well studied. here we identified a homolog of strica from aedes albopictus, named as aacasps16. aacasps16 encoded a protein containing a putative serine- and threonine-rich prodomain ...201627351972
tnfα impairs rhabdoviral clearance by inhibiting the host autophagic antiviral response.tnfα is a pleiotropic pro-inflammatory cytokine with a key role in the activation of the immune system to fight viral infections. despite its antiviral role, a few viruses might utilize the host produced tnfα to their benefit. some recent reports have shown that anti-tnfα therapies could be utilized to treat certain viral infections. however, the underlying mechanisms by which tnfα can favor virus replication have not been identified. here, a rhabdoviral infection model in zebrafish allowed us t ...201627351838
mapping of human b-cell epitopes of sindbis virus.mosquito-transmitted sindbis virus (sinv) causes fever, skin lesions and musculoskeletal symptoms if transmitted to man. sinv is the prototype virus of genus alphavirus, which includes other arthritogenic viruses such as chikungunya virus (chikv) and ross river virus (rrv) that cause large epidemics with a considerable public health burden. until now the human b-cell epitopes have been studied for chikv and rrv, but not for sinv. to identify the b-cell epitopes in sinv-infection, we synthetised ...201627339177
host translation shutoff mediated by non-structural protein 2 is a critical factor in the antiviral state resistance of venezuelan equine encephalitis virus.most previous studies of interferon-alpha/beta (ifn-α/β) response antagonism by alphaviruses have focused upon interruption of ifn-α/β induction and/or receptor signaling cascades. infection of mice with venezuelan equine encephalitis alphavirus (veev) or sindbis virus (sinv) induces serum ifn-α/β, that elicits a systemic antiviral state in uninfected cells successfully controlling sinv but not veev replication. furthermore, veev replication is more resistant than that of sinv to a pre-existing ...201627318152
mosquito saliva increases endothelial permeability in the skin, immune cell migration, and dengue pathogenesis during antibody-dependent enhancement.dengue remains the most prevalent arthropod-borne viral disease in humans. while probing for blood vessels, aedes aegypti and ae. albopictus mosquitoes transmit the four serotypes of dengue virus (denv1-4) by injecting virus-containing saliva into the skin. even though arthropod saliva is known to facilitate transmission and modulate host responses to other pathogens, the full impact of mosquito saliva on dengue pathogenesis is still not well understood. inoculating mice lacking the interferon-α ...201627310141
alphavirus infection: host cell shut-off and inhibition of antiviral responses.alphaviruses cause debilitating disease in humans and animals and are transmitted by blood-feeding arthropods, typically mosquitoes. with a traditional focus on two models, sindbis virus and semliki forest virus, alphavirus research has significantly intensified in the last decade partly due to the re-emergence and dramatic expansion of chikungunya virus in asia, europe, and the americas. as a consequence, alphavirus-host interactions are now understood in much more molecular detail, and importa ...201627294951
receptor-targeted nipah virus glycoproteins improve cell-type selective gene delivery and reveal a preference for membrane-proximal cell attachment.receptor-targeted lentiviral vectors (lvs) can be an effective tool for selective transfer of genes into distinct cell types of choice. moreover, they can be used to determine the molecular properties that cell surface proteins must fulfill to act as receptors for viral glycoproteins. here we show that lvs pseudotyped with receptor-targeted nipah virus (niv) glycoproteins effectively enter into cells when they use cell surface proteins as receptors that bring them closely enough to the cell memb ...201627281338
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
capturing and concentrating adenovirus using magnetic anionic nanobeads.we recently demonstrated how various enveloped viruses can be efficiently concentrated using magnetic beads coated with an anionic polymer, poly(methyl vinyl ether-maleic anhydrate). however, the exact mechanism of interaction between the virus particles and anionic beads remains unclear. to further investigate whether these magnetic anionic beads specifically bind to the viral envelope, we examined their potential interaction with a nonenveloped virus (adenovirus). the beads were incubated with ...201627274228
a tale of two rnas during viral infection: how viruses antagonize mrnas and small non-coding rnas in the host cell.viral infection initiates an array of changes in host gene expression. many viruses dampen host protein expression and attempt to evade the host anti-viral defense machinery. host gene expression is suppressed at several stages of host messenger rna (mrna) formation including selective degradation of translationally competent messenger rnas. besides mrnas, host cells also express a variety of noncoding rnas, including small rnas, that may also be subject to inhibition upon viral infection. in th ...201627271653
host-feeding patterns of mosquito species in germany.mosquito-borne pathogens are of growing importance in many countries of europe including germany. at the same time, the transmission cycles of most mosquito-borne pathogens (e.g. viruses or filarial parasites) are not completely understood. there is especially a lack of knowledge about the vector capacity of the different mosquito species, which is strongly influenced by their host-feeding patterns. while this kind of information is important to identify the relevant vector species, e.g. to dire ...201627259984
a new defective helper rna to produce recombinant sindbis virus that infects neurons but does not propagate.recombinant sindbis viruses are important tools in neuroscience because they combine rapid and high transgene expression with a capacity to carry large transgenes. currently, two packaging systems based on the defective helper (dh) rnas dh(26s)5'sin and dh-bb(trna;te12) are available for generating recombinant sindbis virus that is neurotropic (able to infect neurons and potentially other cells). both systems produce a fraction of viral particles that can propagate beyond the primary infected ne ...201627252627
forced complementation between subgenomic rnas: does human immunodeficiency type 1 virus reverse transcription occur in viral core, cytoplasm, or early endosome?although the process of reverse transcription is well elucidated, it remains unclear if viral core disruption provides a more cellular or viral milieu for hiv-1 reverse transcription. we have devised a method to require mixing of viral cores or core constituents to produce infectious progeny virus by a bipartite subgenomic rna (sgrna) system, in which hiv-1 cplt_r/u5/gag/δpol and nfl sgrnas are complementary to each other and when together can complete viral reverse transcription. only the heter ...201527239643
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
a glance at subgenomic flavivirus rnas and micrornas in flavivirus infections.the family flaviviridae comprises a wide variety of viruses that are distributed worldwide, some of which are associated with high rates of morbidity and mortality. there are neither vaccines nor antivirals for most flavivirus infections, reinforcing the importance of research on different aspects of the viral life cycle. during infection, cytoplasmic accumulation of rna fragments mainly originating from the 3' utrs, which have been designated subgenomic flavivirus rnas (sfrnas), has been detect ...201627233361
adapting the stress response: viral subversion of the mtor signaling pathway.the mammalian target of rapamycin (mtor) is a central regulator of gene expression, translation and various metabolic processes. multiple extracellular (growth factors) and intracellular (energy status) molecular signals as well as a variety of stressors are integrated into the mtor pathway. viral infection is a significant stress that can activate, reduce or even suppress the mtor signaling pathway. consequently, viruses have evolved a plethora of different mechanisms to attack and co-opt the m ...201627231932
suppression of usp18 potentiates the anti-hbv activity of interferon alpha in hepg2.2.15 cells via jak/stat signaling.ubiquitin-specific protease 18 (usp18, also known as ubp43) has both interferon stimulated gene 15 (isg15) dependent and isg15-independent functions. by silencing the expression of usp18 in hepg2.2.15 cells, we studied the effect of usp18 on the anti-hbv activity of ifn-f and demonstrated that knockdown of usp18 significantly inhibited the hbv expression and increased the expression of isgs. levels of hepatitis b virus surface antigen (hbsag), hepatitis b virus e antigen (hbeag), hbv dna and int ...201627227879
hepatitis c virus infection induces autophagy as a prosurvival mechanism to alleviate hepatic er-stress response.hepatitis c virus (hcv) infection frequently leads to chronic liver disease, liver cirrhosis and hepatocellular carcinoma (hcc). the molecular mechanisms by which hcv infection leads to chronic liver disease and hcc are not well understood. the infection cycle of hcv is initiated by the attachment and entry of virus particles into a hepatocyte. replication of the hcv genome inside hepatocytes leads to accumulation of large amounts of viral proteins and rna replication intermediates in the endopl ...201627223299
antagonism of the sodium-potassium atpase impairs chikungunya virus infection.chikungunya virus (chikv) is a reemerging alphavirus that has caused epidemics of fever, arthralgia, and rash worldwide. there are currently no licensed vaccines or antiviral therapies available for the prevention or treatment of chikv disease. we conducted a high-throughput, chemical compound screen that identified digoxin, a cardiac glycoside that blocks the sodium-potassium atpase, as a potent inhibitor of chikv infection. treatment of human cells with digoxin or a related cardiac glycoside, ...201627222471
alphavirus restriction by ifitm proteins.interferon inducible transmembrane proteins (ifitms) are broad-spectrum antiviral factors. in cell culture the entry of many enveloped viruses, including orthomyxo-, flavi-, and filoviruses, is inhibited by ifitms, though the mechanism(s) involved remain unclear and may vary between viruses. we demonstrate that sindbis and semliki forest virus (sfv), which both use endocytosis and acid-induced membrane fusion in early endosomes to infect cells, are restricted by the early endosomal ifitm3. the l ...201627219333
a de novo transcriptome analysis shows that modulation of the jak-stat signaling pathway by salmonid alphavirus subtype 3 favors virus replication in macrophage/dendritic-like to-cells.the janus kinase (jak) and signaling transducer activator of transcription (stat) pathway mediates the signaling of genes required for cellular development and homeostasis. to elucidate the effect of type i ifn on the jak/stat pathway in salmonid alphavirus subtype 3 (sav3) infected macrophage/dendritic like to-cells derived from atlantic salmon (salmo salar l) headkidney leukocytes, we used a differential transcriptome analysis by rna-seq and the kyoto encyclopedia of genes and genomes (keggs) ...201627215196
glial cell activation, recruitment, and survival of b-lineage cells following mcmv brain infection.chemokines produced by reactive glia drive migration of immune cells and previous studies from our laboratory have demonstrated that cd19(+) b cells infiltrate the brain. in this study, in vivo and in vitro experiments investigated the role of reactive glial cells in recruitment and survival of b-lineage cells in response to (murine cytomegalovirus) mcmv infection.201627207308
the modulation of apoptotic pathways by gammaherpesviruses.apoptosis or programmed cell death is a tightly regulated process fundamental for cellular development and elimination of damaged or infected cells during the maintenance of cellular homeostasis. it is also an important cellular defense mechanism against viral invasion. in many instances, abnormal regulation of apoptosis has been associated with a number of diseases, including cancer development. following infection of host cells, persistent and oncogenic viruses such as the members of the gamma ...201627199919
isg12a restricts hepatitis c virus infection through the ubiquitination-dependent degradation pathway.interferons (ifns) restrict various kinds of viral infection via induction of hundreds of ifn-stimulated genes (isgs), while the functions of the majority of isgs are broadly unclear. here, we show that a high-ifn-inducible gene, isg12a (also known as ifi27), exhibits a nonapoptotic antiviral effect on hepatitis c virus (hcv) infection. viral ns5a protein is targeted specifically by isg12a, which mediates ns5a degradation via a ubiquitination-dependent proteasomal pathway. k374r mutation in ns5a ...201627194766
isg15 deficiency and increased viral resistance in humans but not mice.isg15 is an interferon (ifn)-α/β-induced ubiquitin-like protein. it exists as a free molecule, intracellularly and extracellularly, and conjugated to target proteins. studies in mice have demonstrated a role for isg15 in antiviral immunity. by contrast, human isg15 was shown to have critical immune functions, but not in antiviral immunity. namely, free extracellular isg15 is crucial in ifn-γ-dependent antimycobacterial immunity, while free intracellular isg15 is crucial for usp18-mediated downre ...201627193971
making bunyaviruses talk: interrogation tactics to identify host factors required for infection.the identification of host cellular genes that act as either proviral or antiviral factors has been aided by the development of an increasingly large number of high-throughput screening approaches. here, we review recent advances in which these new technologies have been used to interrogate host genes for the ability to impact bunyavirus infection, both in terms of technical advances as well as a summary of biological insights gained from these studies.201627187446
inflammatory and oxidative stress in rotavirus infection.rotaviruses are the single leading cause of life-threatening diarrhea affecting children under 5 years of age. rotavirus entry into the host cell seems to occur by sequential interactions between virion proteins and various cell surface molecules. the entry mechanisms seem to involve the contribution of cellular molecules having binding, chaperoning and oxido-reducing activities. it appears to be that the receptor usage and tropism of rotaviruses is determined by the species, cell line and rotav ...027175349
gain-of-function mutations in protein kinase cα (pkcα) may promote synaptic defects in alzheimer's disease.alzheimer's disease (ad) is a progressive dementia disorder characterized by synaptic degeneration and amyloid-β (aβ) accumulation in the brain. through whole-genome sequencing of 1345 individuals from 410 families with late-onset ad (load), we identified three highly penetrant variants in prkca, the gene that encodes protein kinase cα (pkcα), in five of the families. all three variants linked with load displayed increased catalytic activity relative to wild-type pkcα as assessed in live-cell im ...201627165780
dengue virus reporter replicon is a valuable tool for antiviral drug discovery and analysis of virus replication mechanisms.dengue, the most prevalent arthropod-borne viral disease, is caused by the dengue virus (denv), a member of the flaviviridae family, and is a considerable public health threat in over 100 countries, with 2.5 billion people living in high-risk areas. however, no specific antiviral drug or licensed vaccine currently targets denv infection. the replicon system has all the factors needed for viral replication in cells. since the development of replicon systems, transient and stable reporter replicon ...201627164125
detection of sindbis and inkoo virus rna in genetically typed mosquito larvae sampled in northern sweden.mosquito-borne viruses have a widespread distribution across the globe and are known to pose serious threats to human and animal health. the maintenance and dissemination of these viruses in nature are driven through horizontal and vertical transmission. in the temperate climate of northern sweden, there is a dearth of knowledge on whether mosquito-borne viruses that occur are transmitted transovarially. to gain a better understanding of mosquito-borne virus circulation and maintenance, mosquito ...201627159120
widespread 3'-end uridylation in eukaryotic rna viruses.rna 3' uridylation occurs pervasively in eukaryotes, but is poorly characterized in viruses. in this study, we demonstrate that a broad array of rna viruses, including mycoviruses, plant viruses and animal viruses, possess a novel population of rna species bearing nontemplated oligo(u) or (u)-rich tails, suggesting widespread 3' uridylation in eukaryotic viruses. given the biological relevance of 3' uridylation to eukaryotic rna degradation, we propose a conserved but as-yet-unknown mechanism in ...201627151171
the emerging duck flavivirus is not pathogenic for primates and is highly sensitive to mammalian interferon antiviral signaling.flaviviruses pose a significant threat to both animals and humans. recently, a novel flavivirus, duck tembusu virus (dtmuv), was identified to be the causative agent of a serious duck viral disease in asia. its rapid spread, expanding host range, and uncertain transmission routes have raised substantial concerns regarding its potential threats to nonavian hosts, including humans. here, we demonstrate that dtmuv is not pathogenic for nonhuman primates and is highly sensitive to mammal type i inte ...201627147750
suppression of kaposi's sarcoma-associated herpesvirus infection and replication by 5'-amp-activated protein kinase.the host intracellular antiviral restriction factors inhibit viral infection and replication. the 5'-amp-activated protein kinase (ampk) is a cellular energy sensor regulating metabolic homeostasis. activated ampk inhibits the replication of numerous rna viruses but enhances the entry of vaccinia virus. however, the role of ampk in herpesvirus infection is unclear. in this study, we showed that the constitutive ampk activity restricted kaposi's sarcoma-associated herpesvirus (kshv) lytic replica ...201627147746
down-regulation of p21-activated serine/threonine kinase 1 is involved in loss of mesencephalic dopamine neurons.although the roles of p21-activated serine/threonine kinase 1 (pak1) have been reported in some neurodegenerative diseases, details regarding neurodegeneration are still limited. hence, we tried to determine the role of pak1 and molecular mechanisms of neuronal death involved in neurodegeneration.201627121078
oncolytic potency of her-2 retargeted vsv-fh hybrid viruses: the role of receptor ligand affinity.the hybrid oncolytic vesicular stomatitis virus (vsv-fh) deleted for its g glycoprotein and displaying the measles virus (mv) envelope glycoproteins (hemagglutinin h and fusion f) is fusogenic, infects cells via any of the three mv receptors and has potent oncolytic activity against subcutaneous and disseminated myeloma tumors. to tailor vsv-fh as an oncolytic virus for ovarian cancer, we ablated its natural tropism and retargeted the virus by display of a single-chain antibody (scfv) with speci ...201527119107
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
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
identification of critical residues in hepatitis e virus macro domain involved in its interaction with viral methyltransferase and orf3 proteins.hepatitis e virus (hev) is a major cause of hepatitis in normal and organ transplant individuals. hev open reading frame-1 encodes a polypeptide comprising of the viral nonstructural proteins as well as domains of unknown function such as the macro domain (x-domain), v, duf3729 and y. the macro domain proteins are ubiquitously present from prokaryotes to human and in many positive-strand rna viruses, playing important roles in multiple cellular processes. towards understanding the function of th ...201627113483
vertical transmission of indian ocean lineage of chikungunya virus in aedes aegypti and aedes albopictus mosquitoes.the re-emergence of chikungunya (chik) fever in thailand has been caused by a novel lineage of chikungunya virus (chikv) termed the indian ocean lineage (iol). the aedes albopictus mosquito is thought to be a primary vector of chik fever in thailand, whereas ae. aegypti acts as a secondary vector of the virus. the vertical transmission is believed to be a primary means to maintain chikv in nature and may be associated with an increased risk of outbreak. therefore, the goal of this study was to a ...201627108077
the role of climatic and density dependent factors in shaping mosquito population dynamics: the case of culex pipiens in northwestern italy.culex pipiens mosquito is a species widely spread across europe and represents a competent vector for many arboviruses such as west nile virus (wnv), which has been recently circulating in many european countries, causing hundreds of human cases. in order to identify the main determinants of the high heterogeneity in cx. pipiens abundance observed in piedmont region (northwestern italy) among different seasons, we developed a density-dependent stochastic model that takes explicitly into account ...201627105065
infection pattern and transmission potential of chikungunya virus in two new world laboratory-adapted aedes aegypti strains.chikungunya virus (chikv) is an emerging mosquito-borne virus belonging to the togaviridae, which is transmitted to humans by aedes aegypti and ae. albopictus. we describe the infection pattern of chikv in two new world ae. aegypti strains, hwe and orl. both mosquito strains were susceptible to the virus but showed different infection patterns in midguts and salivary glands. even though acquisition of a bloodmeal showed moderate levels of apoptosis in midgut tissue, there was no obvious addition ...201627102548
an updated checklist of mosquito species (diptera: culicidae) from madagascar.an updated checklist of 235 mosquito species from madagascar is presented. the number of species has increased considerably compared to previous checklists, particularly the last published in 2003 (178 species). this annotated checklist provides concise information on endemism, taxonomic position, developmental stages, larval habitats, distribution, behavior, and vector-borne diseases potentially transmitted. the 235 species belong to 14 genera: aedeomyia (3 species), aedes (35 species), anophel ...201627101839
development and validation of a quantitative, one-step, multiplex, real-time reverse transcriptase pcr assay for detection of dengue and chikungunya viruses.dengue virus (denv) and chikungunya virus (chikv) are important human pathogens with common transmission vectors and similar clinical presentations. patient care may be impacted by the misdiagnosis of denv and chikv in areas where both viruses cocirculate. in this study, we have developed and validated a one-step multiplex reverse transcriptase pcr (rt-pcr) to simultaneously detect, quantify, and differentiate between four denv serotypes (pan-denv) and chikungunya virus. the assay uses taqman te ...201627098955
alphavirus-based vaccines.alphavirus vectors based on semliki forest virus, sindbis virus, and venezuelan equine encephalitis virus have been widely applied for vaccine development. naked rna replicons, recombinant viral particles, and layered dna vectors have been subjected to immunization in preclinical animal models with antigens for viral targets and tumor antigens. moreover, a limited number of clinical trials have been conducted in humans. vaccination with alphavirus vectors has demonstrated efficient immune respon ...201627076308
ecological characterization and molecular differentiation of culex pipiens complex taxa and culex torrentium in eastern austria.culex pipiens complex taxa differ in behaviour, ecophysiology and epidemiologic importance. despite their epidemiologic significance, information on genetic diversity, occurrence and seasonal and spatial distribution patterns of the cx. pipiens complex is still insufficient. assessment of seasonal and spatial distribution patterns of culex pipiens forms and their congener cx. torrentium is crucial for the understanding of their vector-pathogen dynamics.201627067139
pilot longitudinal mosquito surveillance study in the danube delta biosphere reserve and the first reports of anopheles algeriensis theobald, 1903 and aedes hungaricus mihályi, 1955 for romania.mosquito-borne viruses (moboviruses) are of growing importance in many countries of europe. in romania and especially in the danube delta biosphere reserve (ddbr), mosquito and mobovirus surveillance are not performed on a regular basis. however, this type of study is crucially needed to evaluate the risk of pathogen transmission, to understand the ecology of emerging moboviruses, or to plan vector control programmes.201627066827
entry sites of venezuelan and western equine encephalitis viruses in the mouse central nervous system following peripheral infection.venezuelan and western equine encephalitis viruses (veev and weev; alphavirus; togaviridae) are mosquito-borne pathogens causing central nervous system (cns) disease in humans and equids. adult cd-1 mice also develop cns disease after infection with veev and weev. adult cd-1 mice infected by the intranasal (i.n.) route, showed that veev and weev enter the brain through olfactory sensory neurons (osns). in this study, we injected the mouse footpad with recombinant weev (mcmillan) or veev (subtype ...201627053560
tick infestation in birds and prevalence of pathogens in ticks collected from different places in germany.the importance of ticks and tick-borne pathogens for human and animal health has been increasing over the past decades. for their transportation and dissemination, birds may play a more important role than wingless hosts. in this study, tick infestation of birds in germany was examined. eight hundred ninety-two captured birds were infested with ticks and belonged to 48 different species, of which blackbirds (turdus merula) and song thrushes (turdus philomelos) were most strongly infested. ground ...201627048511
a simple and efficient in vivo non-viral rna transfection method for labeling the whole axonal tree of individual adult long-range projection neurons.we report a highly efficient, simple, and non-infective method for labeling individual long-range projection neurons (lrpns) in a specific location with enough sparseness and intensity to allow complete and unambiguous reconstructions of their entire axonal tree. the method is based on the "in vivo" transfection of a large rna construct that drives the massive expression of green fluorescent protein. the method combines two components: injection of a small volume of a hyperosmolar nacl solution ...201627047347
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
sirna screen identifies trafficking host factors that modulate alphavirus infection.little is known about the repertoire of cellular factors involved in the replication of pathogenic alphaviruses. to uncover molecular regulators of alphavirus infection, and to identify candidate drug targets, we performed a high-content imaging-based sirna screen. we revealed an actin-remodeling pathway involving rac1, pip5k1- α, and arp3, as essential for infection by pathogenic alphaviruses. infection causes cellular actin rearrangements into large bundles of actin filaments termed actin foci ...201627031835
autophagy negatively regulates transmissible gastroenteritis virus replication.autophagy is an evolutionarily ancient pathway that has been shown to be important in the innate immune defense against several viruses. however, little is known about the regulatory role of autophagy in transmissible gastroenteritis virus (tgev) replication. in this study, we found that tgev infection increased the number of autophagosome-like double- and single-membrane vesicles in the cytoplasm of host cells, a phenomenon that is known to be related to autophagy. in addition, virus replicatio ...201627029407
molecular determinants of alphavirus neuropathogenesis in mice.alphaviruses are enveloped viruses with a positive-stranded rna genome, of the family togaviridae. in mammals and birds they are mosquito-transmitted and are of veterinary and medical importance. they cause primarily two types of disease: encephalitis and polyarthritis. here we review attempts to understand the molecular basis of encephalitis and virulence for the central nervous system (cns) in mouse models. sindbis virus (sinv) was the first virus to be studied in this way. other viruses analy ...201627028153
middle east respiratory syndrome coronavirus ns4b protein inhibits host rnase l activation.middle east respiratory syndrome coronavirus (mers-cov) is the first highly pathogenic human coronavirus to emerge since severe acute respiratory syndrome coronavirus (sars-cov) in 2002. like many coronaviruses, mers-cov carries genes that encode multiple accessory proteins that are not required for replication of the genome but are likely involved in pathogenesis. evasion of host innate immunity through interferon (ifn) antagonism is a critical component of viral pathogenesis. the ifn-inducible ...201627025250
emergence of a large-plaque variant in mice infected with coxsackievirus b3.coxsackieviruses are enteric viruses that frequently infect humans. to examine coxsackievirus pathogenesis, we orally inoculated mice with the coxsackievirus b3 (cvb3) nancy strain. using hela cell plaque assays with agar overlays, we noticed that some fecal viruses generated plaques >100 times as large as inoculum viruses. these large-plaque variants emerged following viral replication in several different tissues. we identified a single amino acid change, n63y, in the vp3 capsid protein that w ...201627025249
the proteasome controls presynaptic differentiation through modulation of an on-site pool of polyubiquitinated conjugates.differentiation of the presynaptic terminal is a complex and rapid event that normally occurs in spatially specific axonal regions distant from the soma; thus, it is believed to be dependent on intra-axonal mechanisms. however, the full nature of the local events governing presynaptic assembly remains unknown. herein, we investigated the involvement of the ubiquitin-proteasome system (ups), the major degradative pathway, in the local modulation of presynaptic differentiation. we found that prote ...027022091
filopodia and viruses: an analysis of membrane processes in entry mechanisms.filopodia are thin, actin rich bundles protruding from cell plasma membranes, serving physiological purposes, such as probing the environment and facilitating cell-to-cell adhesion. recent studies have highlighted that actively polymerized filopodial-protrusions are exploited during virus entry, trafficking, spread, and the development of clinical pathology of viral diseases. these observations have caused a surge in investigation of the key determinants of filopodial induction and their influen ...201627014223
mutagenesis of coronavirus nsp14 reveals its potential role in modulation of the innate immune response.coronavirus (cov) nonstructural protein 14 (nsp14) is a 60-kda protein encoded by the replicase gene that is part of the replication-transcription complex. it is a bifunctional enzyme bearing 3'-to-5' exoribonuclease (exon) and guanine-n7-methyltransferase (n7-mtase) activities. exon hydrolyzes single-stranded rnas and double-stranded rnas (dsrnas) and is part of a proofreading system responsible for the high fidelity of cov replication. nsp14 n7-mtase activity is required for viral mrna cap syn ...201627009949
analysis of the contribution of hemocytes and autophagy to drosophila antiviral immunity.antiviral immunity in the model organism drosophila melanogaster involves the broadly active intrinsic mechanism of rna interference (rnai) and virus-specific inducible responses. here, using a panel of six viruses, we investigated the role of hemocytes and autophagy in the control of viral infections. injection of latex beads to saturate phagocytosis, or genetic depletion of hemocytes, resulted in decreased survival and increased viral titers following infection with cricket paralysis virus (cr ...201627009948
antimicrobial action of compounds from marine seaweed.seaweed produces metabolites aiding in the protection against different environmental stresses. these compounds show antiviral, antiprotozoal, antifungal, and antibacterial properties. macroalgae can be cultured in high volumes and would represent an attractive source of potential compounds useful for unconventional drugs able to control new diseases or multiresistant strains of pathogenic microorganisms. the substances isolated from green, brown and red algae showing potent antimicrobial activi ...201627005637
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
autophagy in plasma cell ontogeny and malignancy.autophagy is a highly conserved pathway that recycles cytosolic material and organelles via lysosomal degradation. once simplistically viewed as a non-selective survival strategy in dire straits, autophagy has emerged as a tightly regulated process ensuring organelle function, proteome plasticity, cell differentiation and tissue homeostasis, with key roles in physiology and disease. selective target recognition, mediated by specific adapter proteins, enables autophagy to orchestrate highly speci ...201626984755
an rna trapping mechanism in alphavirus mrna promotes ribosome stalling and translation initiation.during translation initiation, eukaryotic initiation factor 2 (eif2) delivers the met-trna to the 40s ribosomal subunit to locate the initiation codon (augi) of mrna during the scanning process. stress-induced eif2 phosphorylation leads to a general blockade of translation initiation and represents a key antiviral pathway in mammals. however, some viral mrnas can initiate translation in the presence of phosphorylated eif2 via stable rna stem-loop structures (dlp; downstream loop) located in thei ...201626984530
pseudomonas aeruginosa activates pkc-alpha to invade middle ear epithelial cells.otitis media (om) is a group of complex inflammatory disorders affecting the middle ear which can be acute or chronic. chronic suppurative otitis media (csom) is a form of chronic om characterized by tympanic membrane perforation and discharge. despite the significant impact of csom on human population, it is still an understudied and unexplored research area. csom is a leading cause of hearing loss and life-threatening central nervous system complications. bacterial exposure especially pseudomo ...201626973629
endogenously-expressed nh2-terminus of circumsporozoite protein interferes with sporozoite invasion of mosquito salivary glands.the circumsporozoite protein is the most abundant polypeptide expressed by sporozoites, the malaria parasite stage capable of infecting humans. sporozoite invasion of mosquito salivary glands prior to transmission is likely mediated by a receptor/ligand-like interaction of the parasites with the target tissues, and the amino (nh2)-terminal portion of csp is involved in this interaction but not the tsr region on the carboxyl (c)-terminus. peptides based on the nh2-terminal domain could compete wi ...201626964736
versatile trans-replication systems for chikungunya virus allow functional analysis and tagging of every replicase protein.chikungunya virus (chikv; genus alphavirus, family togaviridae) has recently caused several major outbreaks affecting millions of people. there are no licensed vaccines or antivirals, and the knowledge of the molecular biology of chikv, crucial for development of efficient antiviral strategies, remains fragmentary. chikv has a 12 kb positive-strand rna genome, which is translated to yield a nonstructural (ns) or replicase polyprotein. chikv structural proteins are expressed from a subgenomic rna ...201626963103
human papillomavirus e2 regulates srsf3 (srp20) to promote capsid protein expression in infected differentiated keratinocytes.the human papillomavirus (hpv) life cycle is tightly linked to differentiation of the infected epithelial cell, suggesting a sophisticated interplay between host cell metabolism and virus replication. previously, we demonstrated in differentiated keratinocytes in vitro and in vivo that hpv type 16 (hpv16) infection caused increased levels of the cellular sr splicing factors (srsfs) srsf1 (asf/sf2), srsf2 (sc35), and srsf3 (srp20). moreover, the viral e2 transcription and replication factor that ...201626962216
virus-based nanoparticles as versatile nanomachines.nanoscale engineering is revolutionizing the way we prevent, detect, and treat diseases. viruses have played a special role in these developments because they can function as prefabricated nanoscaffolds that have unique properties and are easily modified. the interiors of virus particles can encapsulate and protect sensitive compounds, while the exteriors can be altered to display large and small molecules in precisely defined arrays. these properties of viruses, along with their innate biocompa ...201526958921
hunting viral receptors using haploid cells.viruses have evolved intricate mechanisms to gain entry into the host cell. identification of host proteins that serve as viral receptors has enabled insights into virus particle internalization, host and tissue tropism, and viral pathogenesis. in this review we discuss the most commonly employed methods for virus receptor discovery, specifically highlighting the use of forward genetic screens in human haploid cells. the ability to generate true knockout alleles at high saturation provides a sen ...201526958914
cytoplasmic rna granules and viral infection.rna granules are dynamic cellular structures essential for proper gene expression and homeostasis. the two principal types of cytoplasmic rna granules are stress granules, which contain stalled translation initiation complexes, and processing bodies (p bodies), which concentrate factors involved in mrna degradation. rna granules are associated with gene silencing of transcripts; thus, viruses repress rna granule functions to favor replication. this article discusses the breadth of viral interact ...026958719
design of a titering assay for lentiviral vectors utilizing direct extraction of dna from transduced cells in microtiter plates.using lentiviral vector products in clinical applications requires an accurate method for measuring transduction titer. for vectors lacking a marker gene, quantitative polymerase chain reaction is used to evaluate the number of vector dna copies in transduced target cells, from which a transduction titer is calculated. immune design previously described an integration-deficient lentiviral vector pseudotyped with a modified sindbis virus envelope for use in cancer immunotherapy (vp02, of the zvex ...201626942209
distal-less induces elemental color patterns in junonia butterfly wings.the border ocellus, or eyespot, is a conspicuous color pattern element in butterfly wings. for two decades, it has been hypothesized that transcription factors such as distal-less (dll) are responsible for eyespot pattern development in butterfly wings, based on their expression in the prospective eyespots. in particular, it has been suggested that dll is a determinant for eyespot size. however, functional evidence for this hypothesis has remained incomplete, due to technical difficulties.201626937287
genomic approaches for understanding dengue: insights from the virus, vector, and host.the incidence and geographic range of dengue have increased dramatically in recent decades. climate change, rapid urbanization and increased global travel have facilitated the spread of both efficient mosquito vectors and the four dengue virus serotypes between population centers. at the same time, significant advances in genomics approaches have provided insights into host-pathogen interactions, immunogenetics, and viral evolution in both humans and mosquitoes. here, we review these advances an ...201626931545
seroprevalence and risk factors of inkoo virus in northern sweden.the mosquito-borne inkoo virus (inkv) is a member of the california serogroup in the family bunyaviridae, genus orthobunyavirus these viruses are associated with fever and encephalitis, although inkv infections are not usually reported and the incidence is largely unknown. the aim of the study was to determine the prevalence of anti-inkv antibodies and associated risk factors in humans living in northern sweden. seroprevalence was investigated using the world health organization monitoring of tr ...201626928830
wolbachia biocontrol strategies for arboviral diseases and the potential influence of resident wolbachia strains in mosquitoes.arboviruses transmitted by mosquitoes are a major cause of human disease worldwide. the absence of vaccines and effective vector control strategies has resulted in the need for novel mosquito control strategies. the endosymbiotic bacterium wolbachia has been proposed to form the basis for an effective mosquito biocontrol strategy. resident strains of wolbachia inhibit viral replication in drosophila fruit flies and induce a reproductive phenotype known as cytoplasmic incompatibility that allows ...201626925368
small rna profiling in dengue virus 2-infected aedes mosquito cells reveals viral pirnas and novel host mirnas.in aedes mosquitoes, infections with arthropod-borne viruses (arboviruses) trigger or modulate the expression of various classes of viral and host-derived small rnas, including small interfering rnas (sirnas), piwi interacting rnas (pirnas), and micrornas (mirnas). viral sirnas are at the core of the antiviral rna interference machinery, one of the key pathways that limit virus replication in invertebrates. besides sirnas, aedes mosquitoes and cells derived from these insects produce arbovirus-d ...201626914027
dynamics of chikungunya virus cell entry unraveled by single-virus tracking in living cells.chikungunya virus (chikv) is a rapidly emerging mosquito-borne human pathogen causing major outbreaks in africa, asia, and the americas. the cell entry pathway hijacked by chikv to infect a cell has been studied previously using inhibitory compounds. there has been some debate on the mechanism by which chikv enters the cell: several studies suggest that chikv enters via clathrin-mediated endocytosis, while others show that it enters independently of clathrin. here we applied live-cell microscopy ...201626912616
bagaza virus inhibits japanese encephalitis & west nile virus replication in culex tritaeniorhynchus & cx. quinquefasciatus mosquitoes.studies have shown that certain flaviviruses influence susceptibility of mosquitoes by inhibiting/enhancing replication of important flaviviruses. hence, a study was designed to determine whether bagaza virus (bagv), a flavivirus isolated from culex tritaeniorhynchus mosquitoes in india, alters susceptibility of cx. tritaeniorhynchus and cx. quinquefasciatus mosquitoes to japanese encephalitis (jev) and west nile viruses (wnv).026905241
presynaptic serotonin 2a receptors modulate thalamocortical plasticity and associative learning.higher-level cognitive processes strongly depend on a complex interplay between mediodorsal thalamus nuclei and the prefrontal cortex (pfc). alteration of thalamofrontal connectivity has been involved in cognitive deficits of schizophrenia. prefrontal serotonin (5-ht)2a receptors play an essential role in cortical network activity, but the mechanism underlying their modulation of glutamatergic transmission and plasticity at thalamocortical synapses remains largely unexplored. here, we show that ...201626903620
emerging role of lipid droplets in aedes aegypti immune response against bacteria and dengue virus.in mammals, lipid droplets (lds) are ubiquitous organelles that modulate immune and inflammatory responses through the production of lipid mediators. in insects, it is unknown whether lds play any role during the development of immune responses. we show that aedes aegypti aag2 cells - an immune responsive cell lineage - accumulates lds when challenged with enterobacter cloacae, sindbis, and dengue viruses. microarray analysis of aag2 challenged with e.cloacae or infected with dengue virus reveal ...201626887863
hippocampal dendritic spines are segregated depending on their actin polymerization.dendritic spines are mushroom-shaped protrusions of the postsynaptic membrane. spines receive the majority of glutamatergic synaptic inputs. their morphology, dynamics, and density have been related to synaptic plasticity and learning. the main determinant of spine shape is filamentous actin. using frap, we have reexamined the actin dynamics of individual spines from pyramidal hippocampal neurons, both in cultures and in hippocampal organotypic slices. our results indicate that, in cultures, the ...201626881098
the presence and seroprevalence of arthropod-borne viruses in nasiriyah governorate, southern iraq: a cross-sectional study.the knowledge on the presence and seroprevalence of arboviruses in iraq is fragmental. to assess the exposure of the population to arbovirus infections in southern iraq, we conducted a serological screening of the most common arbovirus groups using immunofluorescence, hemagglutination inhibition and neutralization tests. serum samples of 399 adult volunteers were collected in nasiriyah, iraq. antibodies were detected against west nile virus (wnv) (11.6%), sandfly-borne sicilian virus serocomplex ...201626880770
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
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