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: Aims The aim of this study was to investigate the predictive ability of paced QRS duration (pQRSd) for heart failure events among patients receiving right ventricular apical pacing (RVAP). Methods and results A total of 194 patients with complete atrioventricular block receiving pacemaker treatment were enrolled and stratified to group 1, pQRSd 〈 160 ms, n = 53; group 2, 160 ≤ pQRSd 〈 190 ms, n = 97; and group 3, pQRSd ≥ 190 ms, n = 44. Study outcomes were heart failure events, changes in pQRSd, and changes in left ventricular ejection fraction (LVEF). During the 3-year follow-up, the incidence of heart failure events was 9.4, 27.8, and 56.8% in groups 1, 2, and 3, respectively ( P 〈 0.001). Among the patients without heart failure events, the pQRSd at 3 years remained longer than that at baseline (162.1 ± 22.6 vs. 160.9 ± 22.1 ms, P 〈 0.05), whereas among patients who experienced heart failure events, the prolonged pQRSd at 3 years seemed more pronounced as compared with baseline (184.1 ± 21.1 vs. 179.8 ± 21 ms, P 〈 0.001). Linear regression demonstrated that a decrease in LVEF was positively correlated with pQRSd over time (relative risk 0.423; P 〈 0.05). The receiver operating charactersitic curve showed that the cut-off value of pQRSd was 165 ms with a sensitivity of 0.789. Conclusion A prolonged pQRSd has a detrimental effect on long-term cardiac function during RVAP in patients with complete atrioventricular block. pQRSd could be a useful predictor to identify patients who are at risk for heart failure events during RVAP.

Description: Mycobacterium tuberculosis disease represents an enormous global health problem, with exceptionally high morbidity and mortality in HIV-seropositive (HIV + ) persons. Alveolar macrophages from HIV + persons demonstrate specific and targeted impairment of critical host cell responses, including impaired M. tuberculosis -mediated tumor necrosis factor (TNF) release and macrophage apoptosis. Vitamin D may promote anti- M. tuberculosis responses through upregulation of macrophage NO, NADPH oxidase, cathelicidin, and autophagy mechanisms, but whether vitamin D promotes anti- M. tuberculosis mechanisms in HIV + macrophages is not known. In the current study, human macrophages exposed to M. tuberculosis demonstrated robust release of TNF, IB degradation, and NF-B nuclear translocation, and these responses were independent of vitamin D pretreatment. In marked contrast, HIV + U1 human macrophages exposed to M. tuberculosis demonstrated very low TNF release and no significant IB degradation or NF-B nuclear translocation, whereas vitamin D pretreatment restored these critical responses. The vitamin D-mediated restored responses were dependent in part on macrophage CD14 expression. Importantly, similar response patterns were observed with clinically relevant human alveolar macrophages from healthy individuals and asymptomatic HIV + persons at high clinical risk of M. tuberculosis infection. Taken together with the observation that local bronchoalveolar lavage fluid (BALF) levels of vitamin D are severely deficient in HIV + persons, the data from this study demonstrate that exogenous vitamin D can selectively rescue impaired critical innate immune responses in vitro in alveolar macrophages from HIV + persons at risk for M. tuberculosis disease, supporting a potential role for exogenous vitamin D as a therapeutic adjuvant in M. tuberculosis infection in HIV + persons.

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 C virus (HCV) infection is a major cause of severe liver disease. Interferon (IFN)/ribavirin treatment remains the standard therapeutic regimen for HCV infection in most countries. IFN-stimulated genes are believed to contribute to antiviral effects. However, emerging evidence suggests that microRNAs (miRNAs), a class of noncoding small RNAs, are involved in the control of viral infection. Here, we systematically profiled the hepatocyte expression of a set of 750 miRNAs in response to alpha interferon (IFN-α) and interleukin-28B (IL-28B) treatments. The anti-HCV activity of differentially expressed miRNAs was evaluated using cell culture-derived HCV in vitro . The results demonstrate that let-7b had a significant anti-HCV effect by inhibiting HCV replication and viral protein translation in human hepatoma cells. In particular, we show that the inhibition of let-7b attenuated the anti-HCV effects of IFN-α and IL-28B. Furthermore, we show that the host factor insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) is a target of let-7b. IGF2BP1 was required for HCV replication, and its expression was downregulated by IFN-α and IL-28B. Deletion of the wild-type seed region of let-7b abolished its antiviral activity. Finally, we demonstrate that other let-7 family miRNAs were able to inhibit HCV and to suppress IGF2BP1 expression. In conclusion, we provide an example of a host miRNA regulated by type I and type III IFNs that inhibits HCV replication and infectivity by targeting host targets. These results highlight the important role of miRNAs in the host antiviral immune response and provide a novel candidate for anti-HCV therapy.

Description: Congenital human cytomegalovirus (HCMV) infection is the most frequent infectious cause of birth defects, primarily neurological disorders. Neural progenitor/stem cells (NPCs) are the major cell type in the subventricular zone and are susceptible to HCMV infection. In culture, the differentiation status of NPCs may change with passage, which in turn may alter susceptibility to virus infection. Previously, only early-passage (i.e., prior to passage 9) NPCs were studied and shown to be permissive to HCMV infection. In this study, NPC cultures derived at different gestational ages were evaluated after short (passages 3 to 6) and extended (passages 11 to 20) in vitro passages for biological and virological parameters (i.e., cell morphology, expression of NPC markers and HCMV receptors, viral entry efficiency, viral gene expression, virus-induced cytopathic effect, and release of infectious progeny). These parameters were not significantly influenced by the gestational age of the source tissues. However, extended-passage cultures showed evidence of initiation of differentiation, increased viral entry, and more efficient production of infectious progeny. These results confirm that NPCs are fully permissive for HCMV infection and that extended-passage NPCs initiate differentiation and are more permissive for HCMV infection. Later-passage NPCs being differentiated and more permissive for HCMV infection suggest that HCMV infection in fetal brain may cause more neural cell loss and give rise to severe neurological disabilities with advancing brain development.

Description: Porcine circovirus type 2 (PCV2) uses autophagy machinery to enhance its replication in PK-15 cells. However, the underlying mechanisms are unknown. By the use of specific inhibitors, RNA interference, and coimmunoprecipitation, we show that PCV2 induces autophagy in PK-15 cells through a pathway involving the kinases AMP-activated protein kinase (AMPK) and extracellular signal-regulated kinase 1/2 (ERK1/2), the tumor suppressor protein TSC2, and the mammalian target of rapamycin (mTOR). AMPK and ERK1/2 positively regulate autophagy through negative control of the mTOR pathway by phosphorylating TSC2 in PCV2-infected PK-15 cells. Thus, PCV2 might induce autophagy via the AMPK/ERK/TSC2/mTOR signaling pathway in the host cells, representing a pivotal mechanism for PCV2 pathogenesis.

Description: A/chicken/Nanjing/908/2009(H11N2) (CK908) was isolated from a live poultry market in Nanjing, China. Using PCR and sequencing analysis, we obtained the complete genome sequences of the CK908 virus. The sequence analysis demonstrated that this H11N2 virus was a novel reassortant AIV whose PB1, PB2, PA, HA, NP, NA, M, and NS genes originated from H9N2, H7N7, H5N2, H11N8, H3N6, H6N2, H1N1, and H5N1, respectively. Knowledge regarding the complete genome sequences of the CK908 virus will be useful for epidemiological surveillance.