Development of therapeutics for treatment of Ebola virus infection

Ebola virus infection can cause Ebola virus disease (EVD). Patients usually show severe symptoms, and the fatality rate can reach up to 90%. No licensed medicine is available. In this review, development of therapeutics for treatment of Ebola virus infection and EVD will be discussed.     

Patchouli virus X, a new potexvirus from Pogostemon clabin

This work describes a potexvirus obtained from patchouli, Pogostemon clabin, collected in São Paulo, Brazil in 1992. The plants showed mosaic and were infected by a potyvirus and a potexvirus. The potexvirus had a host range limited to Amaranthaceae, Solanaceae and Labiatae and was named Patchouli virus X (PatVX). PatVX was not transmitted by scissors pruning, in tobacco seeds or by Myzus nicotinae. The virus was purified and a specific antiserum with a titre great than 1:512 000 in dot‐ELISA was produced. The virus was serologically related to Papaya mosaic virus, Potato virus X, Viola mottle virus, White clover mosaic virus and Lily virus X. It had a coat protein of 21 071 ± 1 010 Mr. as determined by SDS‐PAGE. Immunolabelling tests demonstrated that fibrillar masses in the cytoplasm contain the coat protein. The presence of a dsRNA was detected in PatVX infected plants.     

狂犬病毒鼠脑复壮后的典型形态恢复及感染细胞内的形态发生学

[目的]观察比较鼠脑复壮前后狂犬病毒的形态变化,并观察病毒感染BHK-21细胞后不同时间的形态发生情况.[方法]以保存时间较长的SRV9毒株为原始材料,经乳鼠脑传代复壮后接种BHK-21细胞,浓缩、纯化后观察.[结果](1)未经复壮的病毒中DI粒子占较高比例,典型粒子只占少数,而复壮后典型粒子所占比例升高到病毒粒子总数的90%.(2)感染24h后在细胞浆内可以观察到典型病毒粒子,其数量随着培养时间的延长而增加.带毒传代之后的培养过程中细胞内病毒数量增加不明显.(3)病毒可以在细胞内的空泡膜表面以多种方式成堆出芽.[结论](1)鼠脑复壮可恢复狂犬病毒中典型粒子所占比例.(2)带毒传代1～2次时为狂犬病毒收获的最佳时机.(3)本研究为狂犬病毒的装配机制补充了数据.     

猴T淋巴细胞趋向性病毒1型双抗原夹心ELISA检测试剂盒的研制

目的研制灵敏度和特异性高的检测实验猴血清中T淋巴细胞趋向性病毒-1型（STLV-1型）／E体的双抗原夹心ELISA（dsELISA）检测试剂盒。方法采用经原核表达系统表达并纯化的人T淋巴细胞白血病病毒-1型（HTLV-1型）的Env蛋白作为包被用抗原，建立了检测STLV-1的dsELISA诊断方法。通过优化反应条件和筛选试剂，确定了dsELISA诊断试剂盒的相关条件，并经敏感性、特异性和重复性试验考查该试剂盒质量。结果试剂盒特异性好，批内重复试验变异系数（CV）〈7％，批间重复试验CV〈10％。对200份猴血清进行随机检测，与国际公认的诊断试剂盒（美国BioReliance公司）的符合率为97％。结论本试剂盒可初步应用于临床上实验猴STLV-1型抗体的检测。     

Inhibitory effect of 2,4‐dichlorophenoxyacetic acid on ROS,autophagy formation,and mRNA replication for influenza virus infection

Influenza virus has had a high rate of antigenic shift and drift that causes significant morbidity and mortality in humans and animals. The lack of excellent pharmacological treatment underlines the importance of the development of the novel antiviral drugs. We investigated the anti‐influenza A and B viruses of 2,4‐dichlorophenoxyacetic acid (2,4‐D), which is the synthetic analog to auxin and is used as a popular herbicide in the agricultural practices. 2,4‐D was evaluated using a cytopathic effect reduction method; assay results showed that 2,4‐D possessed strong anti‐influenza A and B viruses inhibiting the formation of a visible cytopathic effect. Influenza viral RNA expression was performed by quantitative real‐time polymerase chain reaction. 2,4‐D also inhibited virus replication in the early stage of influenza virus infection without direct interaction with virus particles. Additionally, 2,4‐D significantly inhibited various factors occur during influenza virus infection as the acidic vesicular formation and reactive oxygen species production. Moreover, 2,4‐D represented no cytotoxicity in normal kidney cell. Therefore, these findings provide an understanding of the mechanism and efficient use of 2,4‐D in pharmacological applications against influenza virus infection.     

Extracellular DNA in aquatic ecosystems may in part be due to phycovirus activity

In a eutrophic lake, a crash of the algal population was followed by a significant increase in the number of virus-like particles (from ca. 1 10⁶ ml^–1 to ca. 26 10⁶ ml^–1), and soon thereafter by an increase of the amount of extracellular DNA (from ca. 20 µg l^–1 to ca. 40 µg l^–1). The same pattern of correlation between decrease of algae and increase of viruses and extracellular DNA could be demonstrated by an in vitro experiment with a Chlorella-virus-system. Lysis of algae by viruses increased both the number of viruses and the amount of DNA in the culture medium. Extracellular DNA mainly consisted of material with a molecular weight below 500 bp.The Chlorella-virus-system is discussed. It could be used as a model-system for studying the dynamics of interaction of viruses and algae in aquatic ecosystems.     

Site-Specific Structural Variations Accompanying Tubular Assembly of the HIV-1 Capsid Protein

The 231-residue capsid (CA) protein of human immunodeficiency virus type 1 (HIV-1) spontaneously self-assembles into tubes with a hexagonal lattice that is believed to mimic the surface lattice of conical capsid cores within intact virions. We report the results of solid-state nuclear magnetic resonance (NMR) measurements on HIV-1 CA tubes that provide new information regarding changes in molecular structure that accompany CA self-assembly, local dynamics within CA tubes, and possible mechanisms for the generation of lattice curvature. This information is contained in site-specific assignments of signals in two- and three-dimensional solid-state NMR spectra, conformation-dependent ¹⁵N and ¹³C NMR chemical shifts, detection of highly dynamic residues under solution NMR conditions, measurements of local variations in transverse spin relaxation rates of amide ¹H nuclei, and quantitative measurements of site-specific ¹⁵N–¹⁵N dipole–dipole couplings. Our data show that most of the CA sequence is conformationally ordered and relatively rigid in tubular assemblies and that structures of the N-terminal domain (NTD) and the C-terminal domain (CTD) observed in solution are largely retained. However, specific segments, including the N-terminal β-hairpin, the cyclophilin A binding loop, the inter-domain linker, segments involved in intermolecular NTD–CTD interactions, and the C-terminal tail, have substantial static or dynamical disorder in tubular assemblies. Other segments, including the 3₁₀-helical segment in CTD, undergo clear conformational changes. Structural variations associated with curvature of the CA lattice appear to be localized in the inter-domain linker and intermolecular NTD–CTD interface, while structural variations within NTD hexamers, around local 3-fold symmetry axes, and in CTD–CTD dimerization interfaces are less significant.     

No signs of Na+/K+‐ATPase adaptations to an invasive exotic toxic prey in native squamate predators

Invasions by exotic toxic prey, like the release of the South American cane toad (Bufo (Rhinella) marinus) to the toad‐free Australian continent in 1935, have been shown to result in massive declines in native predator numbers. Due to minor nucleotide mutations of the Na⁺/K⁺‐ATPase gene most Australian squamate predators are highly susceptible to cane toad toxin. However, in spite of this, predators like yellow‐spotted goannas (Varanus panoptes) and red‐bellied black snakes (Pseudechis porhyriacus) still persist in parts of Queensland where they, in some areas, have co‐existed with cane toads for more than 70 years. Here, we show that the amino acids of the Na⁺/K⁺‐ATPase enzyme in the two species do not provide toad toxin resistance, and hence the two Queensland predators are still highly susceptible to cane toad toxin. Both yellow‐spotted goannas and lace monitors (Varanus varius) have, however, been recorded avoiding feeding on cane toads in areas where they co‐exist with this toxic amphibian. Moreover, both varanids have also been shown to learn to avoid feeding on toads when first subjected to conditioned taste aversion. Such behavioural shifts may therefore explain why yellow‐spotted goannas and red‐bellied black snakes still exist in cane toad infested areas of Queensland. The process appears, however, to be unable to rapidly restore varanid populations to pre‐toad population numbers as even after 10 years of co‐existence with cane toads in the Northern Territory, we see no signs of an increase in yellow‐spotted goanna numbers.