Description: Infectious bursal disease (IBD) is an acute, highly contagious, and immunosuppressive avian disease caused by IBD virus (IBDV). Although IBDV-induced immunosuppression has been well established, the underlying exact molecular mechanism for such induction is not very clear. We report here the identification of IBDV VP4 as an interferon suppressor by interaction with the glucocorticoid-induced leucine zipper (GILZ) in host cells. We found that VP4 suppressed the expression of type I interferon in HEK293T cells after tumor necrosis factor alpha (TNF-α) treatment or Sendai virus (SeV) infection and in DF-1 cells after poly(I·C) stimulation. In addition, the VP4-induced suppression of type I interferon could be completely abolished by knockdown of GILZ by small interfering RNA (siRNA). Furthermore, knockdown of GILZ significantly inhibited IBDV growth in host cells, and this inhibition could be markedly mitigated by anti-alpha/beta interferon antibodies in the cell cultures ( P 〈 0.001). Thus, VP4-induced suppression of type I interferon is mediated by interaction with GILZ, a protein that appears to inhibit cell responses to viral infection.

Description: Polyelectrolyte-enhanced ultrafiltration was investigated for rhenium(VII) recovery from aqueous solutions by using polyquaternium-6 (PQ6) as a complexing agent. The effects of the operating parameters on the permeate flux ( J ) and the rhenium rejection coefficient ( R ) were studied. In the process of concentration, J declines slowly and R is about 1. The concentrated solution was used for the decomplexation. It takes 10 min to achieve the decomplexation equilibrium at a chloride ion concentration of 100 mg L –1 . The decomplexation percentage reaches 45.6 %. In the diafiltration process, rhenium is extracted effectively, and the purification of the regenerated PQ6 is satisfactory. The regenerated PQ6 was used to bind rhenium(VII). The binding capacity of the regenerated PQ6 is close to that of fresh PQ6. Polyelectrolyte-enhanced ultrafiltration was originally used to achieve the recovery of rhenium from aqueous solutions with the help of polyquaternium-6. The effects of various operating parameters on the permeate flux and the rhenium rejection coefficient were investigated. The integration of four experiments including concentration, decomplexation, diafiltration and reuse of the regenerated polymer was carried out.

Description: A/duck/Shanghai/28-1/2009(H4N2) (DK28) was isolated from a live poultry market in Shanghai, China. Using PCR and sequencing analysis, we obtained the complete genome sequences of the DK28 virus. The sequence analysis demonstrated that this H4N2 virus was a novel multiple-gene reassortant avian influenza virus (AIV) whose genes originated from H1N1, H1N3, H3N3, H4N2, and H4N6. Knowledge regarding the complete genome sequences of the DK28 virus will be useful for epidemiological surveillance.

Description: Well-characterized promoters are essential tools for metabolic engineering and synthetic biology. In Streptomyces coelicolor , the native kasO p is a temporally expressed promoter strictly controlled by two regulators, ScbR and ScbR2. In this work, first, kasO p was engineered to remove a common binding site of ScbR and ScbR2 upstream of its core region, thus generating a stronger promoter, kasO p 3 . Second, another ScbR binding site internal to the kasO p 3 core promoter region was abolished by random mutation and screening of the mutant library to obtain the strongest promoter, kasO p* (where the asterisk is used to distinguish the engineered promoter from the native promoter). The activities of kasO p* were compared with those of two known strong promoters, ermE p* and SF14p, in three Streptomyces species. kasO p* showed the highest activity at the transcription and protein levels in all three hosts. Furthermore, relative to ermE p* and SF14p, kasO p* was shown to confer the highest actinorhodin production level when used to drive the expression of actII -ORF4 in S. coelicolor . Therefore, kasO p* is a simple and well-defined strong promoter useful for gene overexpression in streptomycetes.

Description: [1]  In this paper, the essential technique of Grad-Shafranov (GS) reconstruction is reformulated into an inverse boundary value problems (IBVPs) for Laplace's equation on a circle by introducing a Hilbert transform between the normal and tangent component of the boundary gradients. It is proved that the specified IBVPs have unique solution, given the known Dirichlet and Neumann conditions on certain arc. Even when the arc is reduced to only one point on the circle, it can be inferred logically that the unique solution still exists there on the remaining circle. New solution approach for the specified IBVP is suggested with the help of the introduced Hilbert transform. An iterated Tikhonov regularization scheme is applied to deal with the ill-posed linear operators appearing in the discretization of the new approach. Numerical experiments highlight the efficiency and robustness of the proposed method. According to linearity of the elliptic operator in GS equation, its solution can be divided into twoparts. One is solved from a semi-linear elliptic equation with an homogeneous Dirichlet boundary condition. The other is solved from the IBVP of Laplace's equation. It is concluded that there exists a unique solution for the so-called elliptic Cauchy problem for the essential technique of GS-reconstruction.

Description: Hepatitis B virus (HBV) has extremely restricted host and hepatocyte tropism. HBV-based vectors could form the basis of novel therapies for chronic hepatitis B and other liver diseases and would also be invaluable for the study of HBV infection. Previous attempts at developing HBV-based vectors encountered low yields of recombinant viruses and/or lack of sufficient infectivity/cargo gene expression in primary hepatocytes, which hampered follow-up applications. In this work, we constructed a novel vector based on a naturally occurring, highly replicative HBV mutant with a 207-bp deletion in the preS1/polymerase spacer region. By applying a novel insertion strategy that preserves the continuity of the polymerase open reading frame (ORF), recombinant HBV (rHBV) carrying protein or small interfering RNA (siRNA) genes were obtained that replicated and were packaged efficiently in cultured hepatocytes. We demonstrated that rHBV expressing a fluorescent reporter (DsRed) is highly infective in primary tree shrew hepatocytes, and rHBV expressing HBV-targeting siRNA successfully inhibited antigen expression from coinfected wild-type HBV. This novel HBV vector will be a powerful tool for hepatocyte-targeting gene delivery, as well as the study of HBV infection.

Description: cis -Acting elements in the viral genome RNA (vRNA) are essential for the translation, replication, and/or encapsidation of RNA viruses. In this study, a novel conserved cis -acting element was identified in the capsid-coding region of mosquito-borne flavivirus. The d ownstream of 5' c yclization s equence (5'CS) p seudo k not (DCS-PK) element has a three-stem pseudoknot structure, as demonstrated by structure prediction and biochemical analysis. Using dengue virus as a model, we show that DCS-PK enhances vRNA replication and that its function depends on its secondary structure and specific primary sequence. Mutagenesis revealed that the highly conserved stem 1 and loop 2, which are involved in potential loop-helix interactions, are crucial for DCS-PK function. A predicted loop 1-stem 3 base triple interaction is important for the structural stability and function of DCS-PK. Moreover, the function of DCS-PK depends on its position relative to the 5'CS, and the presence of DCS-PK facilitates the formation of 5'-3' RNA complexes. Taken together, our results reveal that the cis -acting element DCS-PK enhances vRNA replication by regulating genome cyclization, and DCS-PK might interplay with other cis -acting elements to form a functional vRNA cyclization domain, thus playing critical roles during the flavivirus life cycle and evolution.

Description: Viruses that replicate in the cytoplasm cannot access the host nuclear capping machinery. These viruses have evolved viral methyltransferase(s) to methylate N-7 and 2'- O cap of their RNA; alternatively, they "snatch" host mRNA cap to form the 5' end of viral RNA. The function of 2'- O methylation of viral RNA cap is to mimic cellular mRNA and to evade host innate immune restriction. A cytoplasmic virus defective in 2'- O methylation is replicative, but its viral RNA lacks 2'- O methylation and is recognized and eliminated by the host immune response. Such a mutant virus could be rationally designed as a live attenuated vaccine. Here, we use Japanese encephalitis virus (JEV), an important mosquito-borne flavivirus, to prove this novel vaccine concept. We show that JEV methyltransferase is responsible for both N-7 and 2'- O cap methylations as well as evasion of host innate immune response. Recombinant virus completely defective in 2'- O methylation was stable in cell culture after being passaged for 〉30 days. The mutant virus was attenuated in mice, elicited robust humoral and cellular immune responses, and retained the engineered mutation in vivo . A single dose of immunization induced full protection against lethal challenge with JEV strains in mice. Mechanistically, the attenuation phenotype was attributed to the enhanced sensitivity of the mutant virus to the antiviral effects of interferon and IFIT proteins. Collectively, the results demonstrate the feasibility of using 2'- O methylation-defective virus as a vaccine approach; this vaccine approach should be applicable to other flaviviruses and nonflaviviruses that encode their own viral 2'- O methyltransferases.

Description: In order to understand the differences in the suspended sediment and total dissolved solid (TDS) yield patterns between the glacial and non-glacial catchments at the headwaters of Urumqi River, Northwestern China, water samples were collected from a glacier catchment and an empty cirque catchment within the region, during three melting seasons from 2006 to 2008. These samples were analyzed to estimate suspended sediment and TDS concentrations, fluxes and erosion rates in the two adjoining catchments. There were remarked differences in suspended sediment and TDS yield patterns between the two catchments. Suspended sediment concentrations were controlled mainly by the sediment source, whereas TDS concentrations were primarily related to the hydrologic interaction with soil minerals. Generally, the glacial catchment had much higher suspended sediment and TDS yields, together with higher denudation rates, than the non-glacial catchment. Overall, glacial catchment was mainly dominated by physical denudation process, whereas the non-glacial catchment was jointly influenced by physical and chemical denudation processes. The observed differences in material delivery patterns were mainly controlled by the runoff source and the glacial processes. The melting periods of glacier and snow were typically the most important time for the suspended sediment and TDS yields. Meanwhile, episodic precipitation events could generate disproportionately large yields. Subglacial hydrology dynamics, glaciers pluck and grind processes could affect erodibility, and the large quantities of dust stored on the glacier surface provided additional sources for suspended sediment transport in the glacial catchment. These mechanisms imply that, in response to climate change, the catchment behavior will be modified significantly in this region, in terms of material flux. This article is protected by copyright. All rights reserved.

Description: The emergence of resistance to carbapenems in Pseudomonas aeruginosa can be suppressed by optimizing the administration of meropenem. However, whether the same is true for Acinetobacter baumannii is not fully understood. We assessed the bactericidal activity of meropenem and its potency to suppress the emergence of resistance in A. baumannii with human simulated exposure in an in vitro intravenous-infusion hollow-fiber infection model (HFIM). Two clinical strains of carbapenem-susceptible multidrug-resistant A. baumannii (CS-MDRAB), CSRA24 and CSRA91, were used, and their MICs and mutant prevention concentrations (MPCs) were determined. Six meropenem dosage regimens (0.5, 1.0, or 2.0 g given every 8 h [q8h] with a 0.5-h or 3-h infusion for seven consecutive days) were simulated and then evaluated in the HFIM. Both the total population and resistant subpopulations of the two strains were quantified. Drug concentrations were measured by high-performance liquid chromatography. All dosage regimens, except for the lowest dosage (0.5 g for both the 0.5-h and 3-h infusions), showed 3-log CFU/ml bacterial killing. Dosage regimens of 2.0 g with 0.5-h and 3-h infusions exhibited an obvious bactericidal effect and suppressed resistance. Selective amplification of subpopulations with reduced susceptibility to meropenem was suppressed with a percentage of the dosage interval in which meropenem concentrations exceeded the MPC ( T 〉MPC) of ≥20% or with a ratio of T 〉MPC to the percentage of the dosage interval in which drug concentrations are within the mutant selection window of ≥0.25. Our in vitro data support the use of a high dosage of meropenem (2.0 g q8h) for the treatment of severe infection caused by CS-MDRAB.