Spatial, temporal and molecular dynamics of swine influenza virus-specific CD8 tissue resident memory T cells

For the first time we have defined naïve, central memory, effector memory and differentiated effector porcine CD8 T cells and analyzed their distribution in lymphoid and respiratory tissues after influenza infection or immunization, using peptide-MHC tetramers of three influenza nucleoprotein (NP) epitopes. The hierarchy of response to the three epitopes changes during the response in different tissues. Most NP-specific CD8 T cells in broncho-alveolar lavage (BAL) and lung are tissue resident memory cells (TRM) that express CD69 and downregulate CD45RA and CCR7. NP-specific cells isolated from BAL express genes characteristic of TRM, but gene expression differs at 7, 21 and 63 days post infection. In all tissues the frequency of NP-specific CD8 cells declines over 63 days almost to background levels but is best maintained in BAL. The kinetic of influenza specific memory CD8 T cell in this natural host species differs from that in small animal models

Simultaneous aerosol and intramuscular immunization with influenza vaccine induces powerful protective local T cell and systemic antibody immune responses in pigs

A vaccine providing both powerful Ab and cross-reactive T cell immune responses against influenza viruses would be beneficial for both humans and pigs. In this study, we evaluated i.m., aerosol (Aer), and simultaneous systemic and respiratory immunization (SIM) by both routes in Babraham pigs, using the single cycle candidate influenza vaccine S-FLU. After prime and boost immunization, pigs were challenged with H1N1pdm09 virus. i.m.-immunized pigs generated a high titer of neutralizing Abs but poor T cell responses, whereas Aer induced powerful respiratory tract T cell responses but a low titer of Abs. SIM pigs combined high Ab titers and strong local T cell responses. SIM showed the most complete suppression of virus shedding and the greatest improvement in pathology. We conclude that SIM regimes for immunization against respiratory pathogens warrant further study

Magnitude and kinetics of T cell and antibody responses during H1N1pdm09 infection in inbred Babraham pigs and outbred pigs

We have used the pig, a large natural host animal for influenza with many physiological similarities to humans, to characterize αβ, γδ T cell and antibody (Ab) immune responses to the 2009 pandemic H1N1 virus infection. We evaluated the kinetic of virus infection and associated response in inbred Babraham pigs with identical MHC (Swine Leucocyte Antigen) and compared them to commercial outbred animals. High level of nasal virus shedding continued up to days 4 to 5 post infection followed by a steep decline and clearance of virus by day 9. Adaptive T cell and Ab responses were detectable from days 5 to 6 post infection reaching a peak at 9 to 14 days. γδ T cells produced cytokines ex vivo at day 2 post infection, while virus reactive IFNγ producing γδ T cells were detected from day 7 post infection. Analysis of NP tetramer specific and virus specific CD8 and CD4 T cells in blood, lung, lung draining lymph nodes, and broncho-alveolar lavage (BAL) showed clear differences in cytokine production between these tissues. BAL contained the most highly activated CD8, CD4, and γδ T cells producing large amounts of cytokines, which likely contribute to elimination of virus. The weak response in blood did not reflect the powerful local lung immune responses. The immune response in the Babraham pig following H1N1pdm09 influenza infection was comparable to that of outbred animals. The ability to utilize these two swine models together will provide unparalleled power to analyze immune responses to influenza

Pathogenic variants in SLF2 and SMC5 cause segmented chromosomes and mosaic variegated hyperploidy

Embryonic development is dictated by tight regulation of DNA replication, cell division and differentiation. Mutations in DNA repair and replication genes disrupt this equilibrium, giving rise to neurodevelopmental disease characterized by microcephaly, short stature and chromosomal breakage. Here, we identify biallelic variants in two components of the RAD18-SLF1/2-SMC5/6 genome stability pathway, SLF2 and SMC5, in 11 patients with microcephaly, short stature, cardiac abnormalities and anemia. Patient-derived cells exhibit a unique chromosomal instability phenotype consisting of segmented and dicentric chromosomes with mosaic variegated hyperploidy. To signify the importance of these segmented chromosomes, we have named this disorder Atelís (meaning - incomplete) Syndrome. Analysis of Atelís Syndrome cells reveals elevated levels of replication stress, partly due to a reduced ability to replicate through G-quadruplex DNA structures, and also loss of sister chromatid cohesion. Together, these data strengthen the functional link between SLF2 and the SMC5/6 complex, highlighting a distinct role for this pathway in maintaining genome stability

Illuminating the local: can non-formal institutions be complementary to health system development in Papua New Guinea?

This paper offers a fresh insight into the performance and reform opportunities of the formal health system of Papua New Guinea. A central tenant of this paper is that the historically imposed and continuing top-down nature of the formal health system in PNG is not capitalizing on potentially positive incentives and motivations inherent in the broad range of non-formal institutions that frame the PNG health system.    The paper suggests that an enhanced understanding of these non-formal institutions may provide clues for how the formal system could be reconfigured to better align with the non-formal. The opportunity offered by this approach is to leverage the energy, motivation and legitimacy inherent in non-formal institutions to better buttress or infuse the formal health system. The paper draws on an emerging body of development thinking that recognises that development depends on institutions that are stable, fair, legitimate and flexible enough to reflect political pressures; and that in turn, these kinds of institutions are the product of the interplay of formal and informal institutions. In particular, this theory argues it is when informal institutions are “complementary” to formal institutions that institutions are likely to be most effective. In these situations informal institutions support the formal institution through “filling in gaps” either by addressing contingencies not dealt with in the formal rules and/or by facilitating or creating incentives for individuals to pursue the goals of formal institutions. The paper analyses the PNG health system through the lens of this non-formal institutional framework

Rethinking health sector wide approaches through the lens of aid effectiveness

Sector Wide Approaches (SWAps) are an approach to aid management that aim to support recipient government leadership. Health aid has grown rapidly in the past 20 years and in this time SWAps have become an important health aid delivery approach. However, the empirical evidence is that the performance of health SWAps is, at best, mixed. Outcome and impact benefits of health SWAps are inconclusive, and overall process level performance is poor. The key insight offered by this paper is that a divergence has developed between the underlying theory of change of health SWAps, and their implementation. The paper argues the theory of change underlying SWAps is highly consistent with effective aid, however the practice is not. Health SWAps have been characterised by cumbersome architecture that is partially implemented and used, and which does not facilitate government ownership and commitment to indigenous institutional development. A core stream of the analysis as to why this is the case relates to technical shortcomings within the SWAp architecture itself, however this is not sufficient to understand the reasons for the limited success of SWAps. Rather it is clear there are a range of broader incentives and understandings within both donors and governments that have shaped the implementation of SWAps at a process level. The paper concludes with some suggested actions which may assist in realigning health aid with the underlying SWAp theory of change, whilst taking into account at least some of the incentives and understandings that have cut across SWAp implementation. The essence of the proposed approach is a more realistic understanding of the nature of health systems, combined with a more politically informed set of strategies to support the development of these systems. Image: DFID / flick

Feeding response of Ips paraconfusus to phloem and phloem metabolites of Heterobasidion annosum-inoculated ponderosa pine, Pinus ponderosa

In studies of feeding by the bark beetle, Ips paraconfusus, two pine stilbenes (pinosylvin and pinosylvin methyl ether), ferulic acid glucoside, and enantiomers of the four most common sugars present in ponderosa pine phloem (sucrose, glucose, fructose, and raffinose) did not stimulate or reduce male feeding when assayed on wet α-cellulose with or without stimulatory phloem extractives present. When allowed to feed on wet α-cellulose containing sequential extracts (hexane, methanol, and water) of ponderosa pine phloem, methanol and water extractives stimulated feeding, but hexane extractives did not. Males confined in wet α-cellulose containing aqueous or organic extracts of culture broths derived from phloem tissue and containing the root pathogen, Heterobasidion annosum, ingested less substrate than beetles confined to control preparations. In an assay using logs from uninoculated ponderosa pines, the mean lengths of phloem in the digestive tracts increased as time spent feeding increased. Males confined to the phloem of basal logs cut from ponderosa pines artificially inoculated with H. annosum ingested significantly less phloem than beetles in logs cut from trees that were (combined) mock-inoculated or uninoculated and did not contain the pathogen. However, individual pathogen-containing treatments were not significantly different from uninoculated controls. It was concluded that altered feeding rates are not a major factor which may explain why diseased ponderosa pines are colonized by I. paraconfusus

Responding to cough: Boholano illness classification and resort to care in response to childhood ARI

A qualitative study into the health seeking behaviour of caretakers in response to ARI in children under five years of age was conducted in the province of Bohol, the Philippines. The study was designed to compliment survey data generated from a long running ARI intervention project, specifically to explain behaviours identified as problematic by the project. Results indicate the importance of folk diagnosis as a basis for selection of first resort for care in the management of childhood ARI. A cultural category, piang, was identified as a major factor influencing health seeking behaviour and delay in consulting the biomedical system where serious ARI exists. In addition, caretakers' financial situation and social contacts are important in their decision to seek biomedical assistance and are often implicated in delay in presentation and acting upon referral to hospital