Role of position 627 of PB2 and the multibasic cleavage site of the hemagglutinin in the virulence of H5N1 avian influenza virus in chickens and ducks

Highly pathogenic H5N1 avian influenza viruses have caused major disease outbreaks in domestic and free-living birds with transmission to humans resulting in 59% mortality amongst 564 cases. The mutation of the amino acid at position 627 of the viral polymerase basic-2 protein (PB2) from glutamic acid (E) in avian isolates to lysine (K) in human isolates is frequently found, but it is not known if this change affects the fitness and pathogenicity of the virus in birds. We show here that horizontal transmission of A/Vietnam/1203/2004 H5N1 (VN/1203) virus in chickens and ducks was not affected by the change of K to E at PB2-627. All chickens died between 21 to 48 hours post infection (pi), while 70% of the ducks survived infection. Virus replication was detected in chickens within 12 hours pi and reached peak titers in spleen, lung and brain between 18 to 24 hours for both viruses. Viral antigen in chickens was predominantly in the endothelium, while in ducks it was present in multiple cell types, including neurons, myocardium, skeletal muscle and connective tissues. Virus replicated to a high titer in chicken thrombocytes and caused upregulation of TLR3 and several cell adhesion molecules, which may explain the rapid virus dissemination and location of viral antigen in endothelium. Virus replication in ducks reached peak values between 2 and 4 days pi in spleen, lung and brain tissues and in contrast to infection in chickens, thrombocytes were not involved. In addition, infection of chickens with low pathogenic VN/1203 caused neuropathology, with E at position PB2-627 causing significantly higher infection rates than K, indicating that it enhances virulence in chickens.

Predicting neck pain in Royal Australian Air Force fighter pilots

Objective: Fighter pilots frequently report neck pain and injury, and although risk factors have been suggested, the relationships between risk factors and neck pain have not been quantified. The aim of this study was to identify personal and work behaviors that are significantly associated with neck pain in fighter pilots.
Methods: Eighty-two Royal Australian Air Force fighter pilots were surveyed about their flying experience, neck pain prevalence, and prevention. Multinomial logistic regressions were used to fit models between pilots' neck pain during and after flight and a range of personal and work characteristics.
Results: In-flight neck pain was very weakly, yet positively associated with flight hours. Duration of postflight pain was positively associated with the weekly desktop work hours and the sum of preventative actions taken in flight. The duration pilots were considered temporarily medically unfit for flying was positively associated with pilots' age and their weekly desktop work hours.
Discussion: The risk factors identified by the current study should guide neck pain prevention for fighter pilots. In particular, reducing desktop working hours as well as incorporating specific neck-strengthening exercises and in-flight bracing actions should be considered by agencies to help alleviating neck pain in their pilots

Effects of alterations of primer-binding site sequences on human immunodeficiency virus type 1 replication

The human immunodeficiency virus type 1 genomic RNA primer-binding site (PBS) sequence comprises 18 nucleotides which are complementary to those at the 3' end of the replication initiation primer tRNA(3Lys). To investigate the role of the PBS in viral replication, we either deleted the original wild-type PBS (complementary to tRNA(3Lys) or replaced it with DNA sequences complementary to either tRNA(1,2Lys) or tRNA(Phe). Transfection of COS cells with such molecular constructs yielded similar levels of viral progeny that were indistinguishable with regard to viral proteins and tRNA content. Virus particles derived from PBS-deleted molecular clones were noninfectious for MT-4, Jurkat, and CEM-T4 cells. However, infectious viruses were derived from constructs in which the PBS had been altered to sequences complementary to either tRNA(1,2Lys) or tRNA(Phe), although mutated forms showed significant lags in replication efficiency in comparison with wild types. Molecular analysis of reverse-transcribed DNA in cells infected by the mutated viruses indicated that both tRNA(1,2Lys) and tRNA(Phe) could function as primers for reverse transcription during the early stages of infection. Sequencing of full-length proviral DNA, obtained 6 days after infection, revealed the mutated PBS, indicating that a complete cycle of reverse transcription had occurred. During subsequent rounds of infection, reversion of the mutated PBS to wild-type sequences was observed, accompanied by increased production of viral gene products. Reversion to wild-type PBS sequences was confirmed both by specific PCR analysis, using distinct primer pairs, and by direct sequencing of amplified segments. We also performed endogenous in vitro reverse transcription experiments in which synthesis of minus-strand strong-stop viral DNA was primed from a synthetic RNA template containing a PBS complementary to various tRNA isoacceptors. These results showed that tRNA(3Lys) was a much more efficient primer of such reactions than either tRNA(1,2Lys) or tRNA(Phe).

Proteolytic processing of the P2/nucleocapsid cleavage site is critical for human immunodeficiency virus type 1 RNA dimer maturation

Differences in virion RNA dimer stability between mature and protease-defective (immature) forms of human immunodeficiency virus type 1 (HIV-1) suggest that maturation of the viral RNA dimer is regulated by the proteolytic processing of the HIV-1 Gag and Gag-Pol precursor proteins. However, the proteolytic processing of these proteins occurs in several steps denoted primary, secondary, and tertiary cleavage events and, to date, the processing step associated with formation of stable HIV-1 RNA dimers has not been identified. We show here that a mutation in the primary cleavage site (p2/nucleocapsid [NC]) hinders formation of stable virion RNA dimers, while dimer stability is unaffected by mutations in the secondary (matrix/capsid [CA], p1/p6) or a tertiary cleavage site (CA/p2). By introducing mutations in a shared cleavage site of either Gag or Gag-Pol, we also show that the cleavage of the p2/NC site in Gag is more important for dimer formation and stability than p2/NC cleavage in Gag-Pol. Electron microscopy analysis of viral particles shows that mutations in the primary cleavage site in Gag but not in Gag-Pol inhibit viral particle maturation. We conclude that virion RNA dimer maturation is dependent on proteolytic processing of the primary cleavage site and is associated with virion core formation.

One step multifunctional micropatterning of surfaces using asymmetric glow discharge plasma polymerization

Micropatterning of surfaces with varying chemical, physical and topographical properties usually requires a number of fabrication steps. Herein, we describe a micropatterning technique based on plasma enhanced chemical vapour deposition (PECVD) that deposits both protein resistant and protein repellent surface chemistries in a single step. The resulting multifunctional, selective surface chemistries are capable of spatially controlled protein adhesion, geometric confinement of cells and the site specific confinement of enzyme mediated peptide self-assembly.

The thermal stability and structural site-distribution of Eu3+ ions in the red-emitting phosphors Ca9Eu2W4O24 and Sr9Eu2W4O24

The red-emitting phosphors Ca₉Eu₂W₄O₂₄ and Sr₉Eu₂W₄O₂₄ were synthesized by the solid-state reaction method. The crystal phases were characterized by X-ray powder diffraction. The photoluminescence excitation and emission spectra were investigated. The luminescence excitation and emission spectra confirm that the phosphors are efficiently excited by near UV light. The dependence of luminescence intensities on the heating temperatures was investigated. The Ca₉Eu₂W₄O₂₄ phosphor exhibits higher thermal stability than that of Sr₉Eu₂W₄O₂₄. The crystallographic sites for Eu³⁺ ions in Ca₉Eu₂W₄O₂₄ and Sr₉Eu₂W₄O₂₄ are investigated by the site-selective excitation spectra in the ⁵D₀ → ⁷F₀ wavelength region. It is identified that the Eu³⁺ ions occupy only M sites (statistically occupied by 0.5Eu and 0.5Ca) in Ca₉Eu₂W₄O₂₄ and, however, the Eu³⁺ ions can substitute both M sites (Eu³⁺ + Sr²⁺) and Sr²⁺ sites in Sr₉Eu₂W₄O₂₄. The luminescence spectra and the thermal stability are discussed on the basis of the crystal structure, Eu³⁺ site-distributions and the energy transfer.

Use of moral theory to analyse the ethical codes of built environment professional organisations: A case study of the Royal Institution of Chartered Surveyors

Purpose – The purpose of this paper is to discuss the use of moral theory as a philosophical analytical framework for built environment organisations' ethical codes of practice. The identified moral theories under consideration are “deontology”, “consequentialism” and “virtue ethics”.

Design/methodology/approach – The paper uses a case study to examine the use of moral theory to explain the ethical codes of practice of built environment professional organisations. The chosen organisation is the Royal Institution of Chartered Surveyors (RICS). The approach for conducting the case study is through semi-structured interviews with experienced RICS members which gather views on the application of moral theory to explain the RICS ethical principles.

Findings – The case study revealed that there are mixed views on the use of moral theory to explain the RICS code of practice. The general view is that deontology is the most suitable theory to explain the fact that the work or process has been undertaken correctly. On the other hand, there is also a view amongst senior professionals that virtue ethics is most appropriate as it addresses the importance of both the correct “result” and the correct “process”.

Research limitations/implications – The paper uses a case study approach to examine the ethical code of one built environment professional organisation. This research does not therefore claim empirical generalisation but instead provides illustrations on the use of moral theory to explain the code of practice of a built environment professional organisation. The paper is based on a series of interviews. The findings should be understood as the aggregated opinions of the interviewees.

Originality/value – The paper makes an original contribution to existing literature on the theoretical analysis of codes of practice for built environment professional organisations. It describes research which is the first to use moral theory as a framework for analysing rules of conduct of built environment professional organisations.

Metabolite profiles in the anterior cingulate cortex of depressed patients differentiate those taking N-acetyl-cysteine versus placebo

Background: Increased oxidative stress is thought to contribute to the pathophysiology of major depressive disorder (MDD), which is in part due to diminished levels of glutathione, the primary anti-oxidant of the brain. Oral administration of N-acetyl-cysteine (NAC) replenishes glutathione and has therefore been shown to reduce depressive symptoms. Proton magnetic spectroscopy (1H-MRS) that allows quantification of brain metabolites pertinent to both MDD and oxidative biology may provide some novel insights into the neurobiological effects of NAC, and in particular metabolite concentrations within the anterior cingulate cortex (ACC) are likely to be important given the key role of this region in the regulation of affect.

Objective: The aim of this study was to determine whether the metabolite profile of the ACC in MDD patients predicts treatment with adjunctive NAC versus placebo.

Methods: This study was nested within a multicentre, randomized, double-blind, placebo-controlled study of MDD participants treated with adjunctive NAC. Participants (n = 76) from one site completed the spectroscopy component at the end of treatment (12 weeks). Spectra from a single-voxel in the ACC were acquired and absolute concentrations of glutamate (Glu), glutamate-glutamine (Glx), N-acetyl-aspartate (NAA) and myo-inositol (mI) were obtained. Binary logistic regression analysis was performed to determine whether metabolite profiles could predict NAC versus placebo group membership.

Results: When predicting group outcome (NAC or placebo), Glx, NAA and mI were a significant model, and had 75% accuracy, while controlling for depression severity and sex. However, the Glu, NAA and mI profile was only predictive at a trend level, with 68.3% accuracy. For both models, the log of the odds of a participant being in the NAC group was positively related to NAA, Glx and Glu levels and negatively related to mI levels.

Conclusion: The finding of higher Glx and NAA levels being predictive of the NAC group provides preliminary support for the putative anti-oxidative role of NAC in MDD.