The Mature Virion of Ectromelia Virus,a Pathogenic Poxvirus,Is Capable of Intrahepatic Spread and Can Serve as a Target for Delayed Therapy

Orthopoxviruses (OPVs), which include the agent of smallpox (variola virus), the zoonotic monkeypox virus, the vaccine and zoonotic species vaccinia virus, and the mouse pathogen ectromelia virus (ECTV), form two types of infectious viral particles: the mature virus (MV), which is cytosolic, and the enveloped virus (EV), which is extracellular. It is believed that MVs are required for viral entry into the host, while EVs are responsible for spread within the host. Following footpad infection of susceptible mice, ECTV spreads lymphohematogenously, entering the liver at 3 to 4 days postinfection (dpi). Afterwards, ECTV spreads intrahepatically, killing the host. We found that antibodies to an MV protein were highly effective at curing mice from ECTV infection when administered after the virus reached the liver. Moreover, a mutant ECTV that does not make EV was able to spread intrahepatically and kill immunodeficient mice. Together, these findings indicate that MVs are sufficient for the spread of ECTV within the liver and could have implications regarding the pathogenesis of other OPVs, the treatment of emerging OPV infections, as well as strategies for preparedness in case of accidental or intentional release of pathogenic OPVs.     

The Golgi Protein ACBD3, an Interactor for Poliovirus Protein 3A,Modulates Poliovirus Replication

We have shown that the circulating vaccine-derived polioviruses responsible for poliomyelitis outbreaks in Madagascar have recombinant genomes composed of sequences encoding capsid proteins derived from poliovaccine Sabin, mostly type 2 (PVS2), and sequences encoding nonstructural proteins derived from other human enteroviruses. Interestingly, almost all of these recombinant genomes encode a nonstructural 3A protein related to that of field coxsackievirus A17 (CV-A17) strains. Here, we investigated the repercussions of this exchange, by assessing the role of the 3A proteins of PVS2 and CV-A17 and their putative cellular partners in viral replication. We found that the Golgi protein acyl-coenzyme A binding domain-containing 3 (ACBD3), recently identified as an interactor for the 3A proteins of several picornaviruses, interacts with the 3A proteins of PVS2 and CV-A17 at viral RNA replication sites, in human neuroblastoma cells infected with either PVS2 or a PVS2 recombinant encoding a 3A protein from CV-A17 [PVS2-3A(CV-A17)]. The small interfering RNA-mediated downregulation of ACBD3 significantly increased the growth of both viruses, suggesting that ACBD3 slowed viral replication. This was confirmed with replicons. Furthermore, PVS2-3A(CV-A17) was more resistant to the replication-inhibiting effect of ACBD3 than the PVS2 strain, and the amino acid in position 12 of 3A was involved in modulating the sensitivity of viral replication to ACBD3. Overall, our results indicate that exchanges of nonstructural proteins can modify the relationships between enterovirus recombinants and cellular interactors and may thus be one of the factors favoring their emergence.     

A Second CRM1-Dependent Nuclear Export Signal in the Influenza A Virus NS2 Protein Contributes to the Nuclear Export of Viral Ribonucleoproteins

Influenza A virus NS2 protein, also called nuclear export protein (NEP), is crucial for the nuclear export of viral ribonucleoproteins. However, the molecular mechanisms of NEP mediation in this process remain incompletely understood. A leucine-rich nuclear export signal (NES2) in NEP, located at the predicted N2 helix of the N-terminal domain, was identified in the present study. NES2 was demonstrated to be a transferable NES, with its nuclear export activity depending on the nuclear export receptor chromosome region maintenance 1 (CRM1)-mediated pathway. The interaction between NEP and CRM1 is coordinately regulated by both the previously reported NES (NES1) and now the new NES2. Deletion of the NES1 enhances the interaction between NEP and CRM1, and deletion of the NES1 and NES2 motifs completely abolishes this interaction. Moreover, NES2 interacts with CRM1 in the mammalian two-hybrid system. Mutant viruses containing NES2 alterations generated by reversed genetics exhibit reduced viral growth and delay in the nuclear export of viral ribonucleoproteins (vRNPs). The NES2 motif is highly conserved in the influenza A and B viruses. The results demonstrate that leucine-rich NES2 is involved in the nuclear export of vRNPs and contributes to the understanding of nucleocytoplasmic transport of influenza virus vRNPs.     

Functional and Structural Characterization of Neutralizing Epitopes of Measles Virus Hemagglutinin Protein

Effective vaccination programs have dramatically reduced the number of measles-related deaths globally. Although all the available data suggest that measles eradication is biologically feasible, a structural and biochemical basis for the single serotype nature of measles virus (MV) remains to be provided. The hemagglutinin (H) protein, which binds to two discrete proteinaceous receptors, is the major neutralizing target. Monoclonal antibodies (MAbs) recognizing distinct epitopes on the H protein were characterized using recombinant MVs encoding the H gene from different MV genotypes. The effects of various mutations on neutralization by MAbs and virus fitness were also analyzed, identifying the location of five epitopes on the H protein structure. Our data in the present study demonstrated that the H protein of MV possesses at least two conserved effective neutralizing epitopes. One, which is a previously recognized epitope, is located near the receptor-binding site (RBS), and thus MAbs that recognize this epitope blocked the receptor binding of the H protein, whereas the other epitope is located at the position distant from the RBS. Thus, a MAb that recognizes this epitope did not inhibit the receptor binding of the H protein, rather interfered with the hemagglutinin-fusion (H-F) interaction. This epitope was suggested to play a key role for formation of a higher order of an H-F protein oligomeric structure. Our data also identified one nonconserved effective neutralizing epitope. The epitope has been masked by an N-linked sugar modification in some genotype MV strains. These data would contribute to our understanding of the antigenicity of MV and support the global elimination program of measles.     

Macrophage migration inhibitory factor in mud crab Scylla paramamosain: Molecular cloning, expression profiles in various tissues and under Vibrio challenge

As one of the first found cytokines, macrophage migration inhibitory factor (MIF) plays an important role in several physiological processes in crabs. In this study, a full-length MIF cDNA (GenBank accession number: JX131610) from mud crab Scylla paramamosain (Sp) was cloned based on a sequence of S. paramamosain cDNA library. The full length of SpMIF was 734 bp consisting of a 363 bp open reading frame encoding the SpMIF, a 120 amino acid peptide chain. The molecular weight of SpMIF was 13.46 kDa with the pI of 6.82. The alignment analysis showed that SpMIF appeared to be closely related to the counterpart from Eriocheir sinensis (68%). Quantitative real-time PCR analysis revealed that SpMIF was highly expressed in hepatopancreas and hemocytes. In addition, the expression level of SpMIF was increased significantly after a 6-h challenge by Vibrio parahaemolyticus (4.00 × 10⁶ CFU/mL), peaked at 8 h, and then declined to the common level in 48 h. This data indicated that SpMIF was cloned successfully, and suggested that it participated in the immune system of mud crabs.     

Microsatellite genetic variation in the Chinese endemic Eucommia ulmoides (Eucommiaceae): implications for conservation

Eucommia ulmoides, a traditional Chinese medicinal plant, is endangered as a consequence of long‐term and widespread harvest in the late 20th century. It has been widely cultivated as a source of herbal medicine and for use in the organic chemical industry in China. In this study, eight microsatellite markers were applied to investigate genetic diversity in E. ulmoides. Three hundred individuals from one semi‐wild population and nine cultivated populations across its main production area were collected. A high level of genetic diversity at population levels (H_E = 0.716) was observed. The highly outcrossed mating system, high longevity of E. ulmoides and seed admixture may be responsible for high genetic variation within populations. A genetic bottleneck was observed in one population. Populations were only slightly differentiated from one another (F_ST = 0.063); this was also supported by AMOVA, which revealed that 94.05% of the total variation resided within populations. This is probably attributable to long‐distance gene flow mediated by the exchange of seeds by local farmers. Implications of these results for the conservation of genetic resources of E. ulmoides are discussed. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 775–785.     

Revision of the section Anillochlamys Jeannel, 1909 (Coleoptera: Leiodidae: Cholevinae: Leptodirini)

The taxonomy of the Iberian Leptodirini species of the section Anillochlamys Jeannel, 1909 has been revised. The proposed classification is based on the study of the genital structures of both sexes, in particular the internal sac of the aedeagus. According to the different models of internal sacs, the following genera, species and subspecies are identified: genus Anillochlamys Jeannel, 1909: A. aurouxi Español, 1965, A. bueni Jeannel, 1909 (= A. avariae Comas, 1977 n.syn.), A. cullelli Lagar, 1978, A. moroderi Bolívar, 1923 (= A. negrei Comas, 1990 n. syn.), A. subtruncatus Jeannel, 1930 (= A. baguenai Jeannel, 1930) and A. tropicus (Abeille, 1881) (= Adelops hispanicus Ehlers, 1893; A. tropicus var. apicalis Jeannel, 1909); genus Paranillochlamys Zariquiey, 1940: P. catalonicus (Jeannel, 1913), P. urgellesi (Español, 1965) and P. velox Zariquiey, 1940 (= P. velox montadai Lagar, 1963 n. syn.); genus Pseudochlamys Comas, 1977: P. raholai (Zariquiey, 1922) (= Anillochlamys raholai luis-bofilli Zariquiey, 1940 n. syn.); genus Spelaeochlamys Dieck, 1870 (= Typhlochlamys Español, 1975 n.syn.): S.bardisai (Español, 1975) (= Typhlochlamys escolai Comas, 1978 n. syn.), S. ehlersi Dieck, 1870 and S. ehlersi verai Comas, 1977 n. stat.     

Ostericum atropurpureum sp. nov. (Apiaceae) from Zhejiang,China

Ostericum atropurpureum G. Y. Li, G. H. Xia & W. Y. Xie (Apiaceae, Apioideae) from Zhejiang, China, is described and illustrated. It is closely related to O. huadongense Z. H. Pan & X. H. Li and O. sieboldii (Miquel) Nakai, but differs in having leaves with 1.5–9.0 cm long petiole, linear bracteoles 6–12 mm long, 5–9 rays, 7–14‐flowered umbellules, dark purple petals, broadly winged dorsal and lateral fruit ribs, 1.0–1.5 mm broad, 3–6 vittae in each furrow and 4–8 on the commissure.     

Application of exogenous substances reduces tobacco-specific nitrosamines content by regulating biosynthesis of nicotine and nitrite in burley tobacco

Tobacco-specific nitrosamines (TSNAs) are carcinogenic chemicals found in tobacco plants. The increasing health consciousness of individuals had led to an increased interest in research on reducing TSNAs content. The aim of this study was to use a pot experiment in which exogenous substances were applied to burley tobacco to dissect the mechanism of TSNAs production. The results indicated that spraying the exogenous substances IAA, NAA, SA and combination thereof on burley tobacco after topping decreased TSNAs content by 2.69–29.4 % in upper leaves and 0.23–39.3 % in middle leaves without affecting total sugar, total nitrogen, potassium and chlorine contents. The application of exogenous substances could down-regulate expression of the NR gene and the activity of the NR enzyme, resulting in less accumulation of the TSNAs precursor nitrite. The exogenous substances significantly reduced nicotine accumulation, which was consistent with low enzyme activities and the down-regulated expressions of genes involved in nicotine biosynthesis, especially significant in the case of quinolinate phosphoribosyltransferase. These results suggested that the application of exogenous substances on burley tobacco after topping could reduce TSNAs content which may be attributed to the regulation of exogenous substances on nitrite and nicotine. This also implies one potential improvement to agronomic practices aimed at controlling the accumulation of TSNAs in burley tobacco.