Immune responses in pigs vaccinated with adjuvanted and non-adjuvanted A(H1N1)pdm/09 influenza vaccines used in human immunization programmes.

Following the emergence and global spread of a novel H1N1 influenza virus in 2009, two A(H1N1)pdm/09 influenza vaccines produced from the A/California/07/09 H1N1 strain were selected and used for the national immunisation programme in the United Kingdom: an adjuvanted split virion vaccine and a non-adjuvanted whole virion vaccine. In this study, we assessed the immune responses generated in inbred large white pigs (Babraham line) following vaccination with these vaccines and after challenge with A(H1N1)pdm/09 virus three months post-vaccination. Both vaccines elicited strong antibody responses, which included high levels of influenza-specific IgG1 and haemagglutination inhibition titres to H1 virus. Immunisation with the adjuvanted split vaccine induced significantly higher interferon gamma production, increased frequency of interferon gamma-producing cells and proliferation of CD4(-)CD8(+) (cytotoxic) and CD4(+)CD8(+) (helper) T cells, after in vitro re-stimulation. Despite significant differences in the magnitude and breadth of immune responses in the two vaccinated and mock treated groups, similar quantities of viral RNA were detected from the nasal cavity in all pigs after live virus challenge. The present study provides support for the use of the pig as a valid experimental model for influenza infections in humans, including the assessment of protective efficacy of therapeutic interventions

Site-directed M2 proton channel inhibitors enable synergistic combination therapy for rimantadine-resistant pandemic influenza.

Pandemic influenza A virus (IAV) remains a significant threat to global health. Preparedness relies primarily upon a single class of neuraminidase (NA) targeted antivirals, against which resistance is steadily growing. The M2 proton channel is an alternative clinically proven antiviral target, yet a near-ubiquitous S31N polymorphism in M2 evokes resistance to licensed adamantane drugs. Hence, inhibitors capable of targeting N31 containing M2 (M2-N31) are highly desirable. Rational in silico design and in vitro screens delineated compounds favouring either lumenal or peripheral M2 binding, yielding effective M2-N31 inhibitors in both cases. Hits included adamantanes as well as novel compounds, with some showing low micromolar potency versus pandemic "swine" H1N1 influenza (Eng195) in culture. Interestingly, a published adamantane-based M2-N31 inhibitor rapidly selected a resistant V27A polymorphism (M2-A27/N31), whereas this was not the case for non-adamantane compounds. Nevertheless, combinations of adamantanes and novel compounds achieved synergistic antiviral effects, and the latter synergised with the neuraminidase inhibitor (NAi), Zanamivir. Thus, site-directed drug combinations show potential to rejuvenate M2 as an antiviral target whilst reducing the risk of drug resistance

Home and Online Management and Evaluation of Blood Pressure (HOME BP) using a digital intervention in poorly controlled hypertension: randomised controlled trial

Objective: The HOME BP (Home and Online Management and Evaluation of Blood Pressure) trial aimed to test a digital intervention for hypertension management in primary care by combining self-monitoring of blood pressure with guided self-management. Design: Unmasked randomised controlled trial with automated ascertainment of primary endpoint. Setting: 76 general practices in the United Kingdom. Participants: 622 people with treated but poorly controlled hypertension (>140/90 mm Hg) and access to the internet. Interventions: Participants were randomised by using a minimisation algorithm to self-monitoring of blood pressure with a digital intervention (305 participants) or usual care (routine hypertension care, with appointments and drug changes made at the discretion of the general practitioner; 317 participants). The digital intervention provided feedback of blood pressure results to patients and professionals with optional lifestyle advice and motivational support. Target blood pressure for hypertension, diabetes, and people aged 80 or older followed UK national guidelines. Main outcome measures: The primary outcome was the difference in systolic blood pressure (mean of second and third readings) after one year, adjusted for baseline blood pressure, blood pressure target, age, and practice, with multiple imputation for missing values. Results: After one year, data were available from 552 participants (88.6%) with imputation for the remaining 70 participants (11.4%). Mean blood pressure dropped from 151.7/86.4 to 138.4/80.2 mm Hg in the intervention group and from 151.6/85.3 to 141.8/79.8 mm Hg in the usual care group, giving a mean difference in systolic blood pressure of −3.4 mm Hg (95% confidence interval −6.1 to −0.8 mm Hg) and a mean difference in diastolic blood pressure of −0.5 mm Hg (−1.9 to 0.9 mm Hg). Results were comparable in the complete case analysis and adverse effects were similar between groups. Within trial costs showed an incremental cost effectiveness ratio of £11 ($15, €12; 95% confidence interval £6 to £29) per mm Hg reduction. Conclusions: The HOME BP digital intervention for the management of hypertension by using self-monitored blood pressure led to better control of systolic blood pressure after one year than usual care, with low incremental costs. Implementation in primary care will require integration into clinical workflows and consideration of people who are digitally excluded. Trial registration: ISRCTN13790648

Investigation of the novel proteins of Influenza B

Influenza B viruses encode two small proteins, NB and BM2. BM2 is translated from segment 7 mRNAs by a mechanism involving sequence complementarity to the 18S ribosomal subunit. The importance of these complementary sequences was tested in the context of infectious virus using a reverse genetic approach. A series of mutations were introduced into this region of segment 7. Recombinant viruses with disrupted 18S complementarity displayed deficiency in BM2 expression in infected cells. The BM2 protein is essential for virus replication because its ion channel activity is required during virion entry to the cell. There is also evidence that the cytoplasmic tail of BM2 is involved in viral assembly. A series of amino acid truncations and substitutions in the BM2 cytoplasmic tail were engineered. Recombinant viruses that lacked more than 5 residues at the carboxyl terminus of the protein were not recovered and key residues in the region -5 to -10 were identified. The influenza B virus RNA segment 6 encodes the neuraminidase protein, NA, as well as NB. NB is a 100 amino acid transmembrane protein with a glycosylated ectodomain. NB is conserved in all natural influenza B virus isolates. Influenza B viruses that lack the NB protein can replicate in cell culture to wild-type levels, However, the deletant viruses showed attenuated growth in complex airway cultures derived from humans and ferrets. In vivo, infected ferrets excreted infectious virus in the nasal wash one day later than for viruses that encode NB. Alterations in the expression of the NA protein were not responsible for the attenuated phenotype shown by NB deletant viruses. The role of the host cell ESCRT pathway or of the interferon-induced tetherin protein in assembly and release of influenza viruses was assessed. No evidence was found for either host pathway in the replication of influenza viruses

Investigation of the novel proteins of influenza B

Influenza B viruses encode two small proteins, NB and BM2. BM2 is translated from segment 7 mRNAs by a mechanism involving sequence complementarity to the 18S ribosomal subunit. The importance of these complementary sequences was tested in the context of infectious virus using a reverse genetic approach. A series of mutations were introduced into this region of segment 7. Recombinant viruses with disrupted 18S complementarity displayed deficiency in BM2 expression in infected cells. The BM2 protein is essential for virus replication because its ion channel activity is required during virion entry to the cell. There is also evidence that the cytoplasmic tail of BM2 is involved in viral assembly. A series of amino acid truncations and substitutions in the BM2 cytoplasmic tail were engineered. Recombinant viruses that lacked more than 5 residues at the carboxyl terminus of the protein were not recovered and key residues in the region -5 to -10 were identified. The influenza B virus RNA segment 6 encodes the neuraminidase protein, NA, as well as NB. NB is a 100 amino acid transmembrane protein with a glycosylated ectodomain. NB is conserved in all natural influenza B virus isolates. Influenza B viruses that lack the NB protein can replicate in cell culture to wild-type levels, However, the deletant viruses showed attenuated growth in complex airway cultures derived from humans and ferrets. In vivo, infected ferrets excreted infectious virus in the nasal wash one day later than for viruses that encode NB. Alterations in the expression of the NA protein were not responsible for the attenuated phenotype shown by NB deletant viruses. The role of the host cell ESCRT pathway or of the interferon-induced tetherin protein in assembly and release of influenza viruses was assessed. No evidence was found for either host pathway in the replication of influenza viruses.EThOS - Electronic Theses Online ServiceBBSRCGBUnited Kingdo

Molecular studies of influenza B virus in the reverse genetics era

Recovery of an infectious virus of defined genetic structure entirely from cDNA and the deduction of information about the virus resulting from phenotypic characterization of the mutant is the process of reverse genetics. This approach has been possible for a number of negative-strand RNA viruses since the recovery of rabies virus in 1994. However, the recovery of recombinant orthomyxoviruses posed a greater challenge due to the segmented nature of the genome. It was not until 1999 that such a system was reported for influenza A viruses, but since that time our knowledge of influenza A virus biology has grown dramatically. Annual influenza epidemics are caused not only by influenza A viruses but also by influenza B viruses. In 2002, two groups reported the successful recovery of influenza B virus entirely from cDNA. This has allowed greater depth of study into the biology of these viruses. This review will highlight the advances made in various areas of influenza B virus biology as a result of the development of reverse genetics techniques for these viruses, including (i) the importance of the non-coding regions of the influenza B virus genome; (ii) the generation of novel vaccine strains; (iii) studies into the mechanisms of drug resistance; (iv) the function(s) of viral proteins, both those analogous to influenza A virus proteins and those unique to influenza B viruses. The information generated by the application of influenza B virus reverse genetics systems will continue to contribute to our improved surveillance and control of human influenza.</p

University of California Press eScholarship editions

Recognized as a major figure in postwar American painting, Richard Diebenkorn (1922-1993) was an artist strongly identified with California but whose work is beloved throughout the United States and the rest of the world. This catalog is the most comprehensive volume on the artist now available. Jane Livingston's extensively researched biographical essay covers Diebenkorn's entire career and concentrates on the artist's inner life and purposes as revealed in his paintings. Ruth Fine deals primarily with the figurative aspect of Diebenkorn's work (1955-67), and John Elderfield concentrates on the Ocean Park period (1967-93). All three authors provide valuable insights based on their personal relationships with the artist and his widow, Phyllis. On both page and canvas, the reader can sense Diebenkorn's complexity and highly self-conscious working methods, as well as his formidable integrity. The Art of Richard Diebenkorn will give readers with an interest in all phases of modernism new thoughts about the relationship between abstraction and representation. Stunningly illustrated, with 192 full-color reproductions, this book is an exhilarating testament to a distinctive American artist

Multiple sources supply eolian mineral dust to the Atlantic sector of coastal Antarctica: Evidence from recent snow layers at the top of Berkner Island ice sheet

The Sr and Nd isotopic composition of dust extracted from recent snow layers at the top of Berkner Island ice sheet (located within the Filchner–Ronne Ice Shelf at the southern end of the Weddell Sea) enables us, for the first time, to document dust provenance in Antarctica outside the East Antarctic Plateau (EAP) where all previous studies based on isotopic fingerprinting were carried out. Berkner dust displays an overall crust-like isotopic signature, characterized by more radiogenic 87Sr/86Sr and much less radiogenic 143Nd/144Nd compared to dust deposited on the EAP during glacial periods. Differences with EAP interglacial dust are not as marked but still significant, indicating that present-day Berkner dust provenance is distinct, at least to some extent, from that of the dust reaching the EAP. The fourteen snow-pit sub-seasonal samples that were obtained span a two-year period (2002–2003) and their dust Sr and Nd isotopic composition reveals that multiple sources are at play over a yearly time period. Southern South America, Patagonia in particular, likely accounts for part of the observed spring/summer dust deposition maxima, when isotopic composition is shifted towards “younger” isotopic signatures. In the spring, possible additional inputs from Australian sources would also be supported by the data. Most of the year, however, the measured isotopic signatures would be best explained by a sustained background supply from putative local sources in East Antarctica, which carry old-crust-like isotopic fingerprints. Whether the restricted East Antarctic ice-free areas produce sufficient eolian material has yet to be substantiated however. The fact that large (> 5 μm) particles represent a significant fraction of the samples throughout the entire time-series supports scenarios that involve contributions from proximal sources, either in Patagonia and/or Antarctica (possibly including snow-free areas in the Antarctic Peninsula and other areas as well). This also indicates that additional dust transport, which does not reach the EAP, must occur at low-tropospheric levels to this coastal sector of Antarctica