Plasmodium falciparum artemisinin resistance: something gained in translation

Small-Saunders et al. uncovered a new facet of artemisinin resistance in Plasmodium in which parasites use a previously underexplored arm of stress response mechanisms. Through altered epitranscriptomic modifications on tRNA, changed translation patterns adapt resistant cells to facilitate entry into a quiescent-like state which provides the parasite an escape from many drugs

A novel method for the genome-wide high resolution analysis of DNA damage

DNA damage occurs via endogenous and exogenous genotoxic agents and compromises a genome’s integrity. Knowing where damage occurs within a genome is crucial to understanding the repair mechanisms which protect this integrity. This paper describes a new development based on microarray technology which uses ultraviolet light induced DNA damage as a paradigm to determine the position and frequency of DNA damage and its subsequent repair throughout the entire yeast genome

Does higher-yielding agriculture mean more environmental harm?

A criticism of the land-sparing approach to preserving biodiversity, by restricting farmland to a smaller, higher-yielding area, is that other impacts are higher in food produced this way. This study aims to investigate the evidence for this based on currently available data and models for greenhouse gas emissions, N, P and soil loss and water use. We asked 25 experts to identify and supply data to plot environmental impact per unit of product against yield for the beef, dairy, wheat and rice sectors. This produced data from modelling and field trials and the lifecycle assessment and field trial literature. The data were modelled statistically to adjust for differences between the studies. Given data limitations, it does not seem that higher yielding agricultural production has higher impacts, often quite the reverse. We ask those conducting field studies to collect data that can definitively answer this question.CCI project code: CCI 06-16-00

Plasmodium berghei K13 mutations mediate in vivo artemisinin resistance that is reversed by proteasome inhibition

The recent emergence of Plasmodium falciparum parasite resistance to the first line antimalarial drug artemisinin is of particular concern. Artemisinin resistance is primarily driven by mutations in the P. falciparum K13 protein, which enhance survival of early ring-stage parasites treated with the artemisinin active metabolite dihydroartemisinin in vitro and associate with delayed parasite clearance in vivo. However, association of K13 mutations with in vivo artemisinin resistance has been problematic due to the absence of a tractable model. Herein, we have employed CRISPR/Cas9 genome editing to engineer selected orthologous P. falciparum K13 mutations into the K13 gene of an artemisinin-sensitive Plasmodium berghei rodent model of malaria. Introduction of the orthologous P. falciparum K13 F446I, M476I, Y493H, and R539T mutations into P. berghei K13 yielded gene-edited parasites with reduced susceptibility to dihydroartemisinin in the standard 24-h in vitro assay and increased survival in an adapted in vitro ring-stage survival assay. Mutant P. berghei K13 parasites also displayed delayed clearance in vivo upon treatment with artesunate and achieved faster recrudescence upon treatment with artemisinin. Orthologous C580Y and I543T mutations could not be introduced into P. berghei, while the equivalents of the M476I and R539T mutations resulted in significant growth defects. Furthermore, a Plasmodium-selective proteasome inhibitor strongly synergized dihydroartemisinin action in these P. berghei K13 mutant lines, providing further evidence that the proteasome can be targeted to overcome artemisinin resistance. Taken together, our findings provide clear experimental evidence for the involvement of K13 polymorphisms in mediating susceptibility to artemisinins in vitro and, most importantly, under in vivo conditions

Immunization with genetically attenuated P52-deficient Plasmodium berghei sporozoites induces a long-lasting effector memory CD8+ T cell response in the liver

Item does not contain fulltextABSTRACT: BACKGROUND: The induction of sterile immunity and long lasting protection against malaria has been effectively achieved by immunization with sporozoites attenuated by gamma-irradiation or through deletion of genes. For mice immunized with radiation attenuated sporozoites (RAS) it has been shown that intrahepatic effector memory CD8+ T cells are critical for protection. Recent studies have shown that immunization with genetically attenuated parasites (GAP) in mice is also conferred by liver effector memory CD8+ T cells. FINDINGS: In this study we analysed effector memory cell responses after immunization of GAP that lack the P52 protein. We demonstrate that immunization with p52-GAP sporozoites also results in a strong increase of effector memory CD8+ T cells, even 6 months after immunization, whereas no specific CD4+ effector T cells response could be detected. In addition, we show that the increase of effector memory CD8+ T cells is specific for the liver and not for the spleen or lymph nodes. CONCLUSIONS: These results indicate that immunization of mice with P. berghei p52-GAP results in immune responses that are comparable to those induced by RAS or GAP lacking expression of UIS3 or UIS4, with an important role implicated for intrahepatic effector memory CD8+ T cells. The knowledge of the mediators of protective immunity after immunization with different GAP is important for the further development of vaccines consisting of genetically attenuated sporozoites

Salivary gland-specific P. berghei reporter lines enable rapid evaluation of tissue-specific sporozoite loads in mosquitoes

Malaria is a life-threatening human infectious disease transmitted by mosquitoes. Levels of the salivary gland sporozoites (sgs), the only mosquito stage infectious to a mammalian host, represent an important cumulative index of Plasmodium development within a mosquito. However, current techniques of sgs quantification are laborious and imprecise. Here, transgenic P. berghei reporter lines that produce the green fluorescent protein fused to luciferase (GFP-LUC) specifically in sgs were generated, verified and characterised. Fluorescence microscopy confirmed the sgs stage specificity of expression of the reporter gene. The luciferase activity of the reporter lines was then exploited to establish a simple and fast biochemical assay to evaluate sgs loads in whole mosquitoes. Using this assay we successfully identified differences in sgs loads in mosquitoes silenced for genes that display opposing effects on P. berghei ookinete/oocyst development. It offers a new powerful tool to study infectivity of P. berghei to the mosquito, including analysis of vector-parasite interactions and evaluation of transmission-blocking vaccines

An Analysis of the Cost of the Abatement of Ammonia Emissions in Irish Agriculture to 2030

peer-reviewedThis analysis quantifies the potential to abate national ammonia (NH3) emissions up to 2030. This report is an updated marginal abatement cost curve (MACC) analysis where Teagasc has quantified the abatement potential of a range of ammonia mitigation measures, as well as their associated costs/benefits (see Lanigan et al. 2015 for previous analysis). The objective of this analysis is to quantify the extent and costs associated with meeting future ammonia emission targets that were negotiated as part of the amended Clean Air Policy Package. The requirement to reduce ammonia emissions is urgent, both in terms of compliance with the National Emissions Ceilings Directive (NECD), and as a principal loss pathway for agricultural nitrogen (N). Improvement of N efficiency is a key focus for improving farm efficiency and sustainability as well as reducing the ammonia, nitrate and greenhouse gas (GHG) footprint of agriculture. This is particularly relevant in the context of the national strategies on the development of the agri-food sector: Food Wise 2025, Ag-food strategy 2030 and Ag-Climatise (currently under development) and the newly unveiled EU Farm to Fork Strategy, which is a part of the European Green Deal. Under the baseline scenario (S1), agricultural ammonia emissions are projected to increase by 9% (without any mitigation) by 2030 relative to 2005 levels. While these increases are small in comparison to the targeted increase in agricultural output, they will provide a major challenge to meeting emissions targets, particularly as agriculture comprises over 99% of national emissions. The analysis presented in this report seeks to quantify the ammonia mitigation potential under likely uptake pathways. This is not an exhaustive analysis of all mitigation measures, but represents an assessment of best available techniques, based on scientific, peer-reviewed research carried out by Teagasc and associated national and international research partners. Indeed, any future changes in the sector or in the national emission inventory calculations will require further analysis of the applicability of ammonia mitigation techniques, particularly in terms of housing and storage but also in the context of other reactive N1 emissions. It should also be noted that some mitigation measures, particularly those related to nitrogen application to soils, could result in either higher greenhouse gas emissions or higher nitrate leaching. Compared to a future where no mitigation measures are deployed to address emissions, by 2030 the average technical abatement2 potential was estimated to be approximately 15.26 kt NH3 at a net cost of €10.86 million per annum. However, it should be noted that the net cost (€10.86 million) is comprised of 6 measures that are cost negative (-€22.21 million) and 7 measures that are cost positive (€33.07) and that some of the cost negative measures are predicated on efficiency gains driven by best management practice adoption (e.g. liming and clover measures with associate chemical N reductions). Amongst the thirteen mitigation measures selected for this analysis, 80% of the mitigation potential can be achieved by the full implementation of the mitigation pathways for protected urea and low emission slurry spreading (LESS) techniques for bovines. It should be stressed that this is an assessment of the maximum abatement potential and realising this level of abatement in practice will be extremely challenging. Any increase in agricultural activity beyond the baseline scenario will increase absolute emissions. The level of mitigation achievable is based on the draft AgClimatise measures any delay or reduction in the uptake of these measures will reduce the mitigation achieved. It must also be ensured that all mitigation measures should, where possible, be synergistic with reductions in greenhouse gas emissions and N loss to water

A systematic search for changing-look quasars in SDSS

CLM acknowledges support from the STFC Consolidated Grant (Ref. St/M001229/1). NPR acknowledges support from the STFC and the Ernest Rutherford Fellowship scheme. KH acknowledges support from STFC grant ST/M001296/1. Funding for the SDSS and SDSS-II has been provided by the Alfred P. Sloan Foundation, the Participating Institutions, the National Science Foundation, the US Department of Energy, the National Aeronautics and Space Administration, the Japanese Monbukagakusho, the Max Planck Society, and the Higher Education Funding Council for England.We present a systematic search for changing-look quasars based on repeat photometry from Sloan Digital Sky Survey (SDSS) and Pan-STARRS1, along with repeat spectra from SDSS and SDSS-III Baryon Oscillation Spectroscopic Survey. Objects with large, |Δg| > 1 mag photometric variations in their light curves are selected as candidates to look for changes in broad emission line (BEL) features. Out of a sample of 1011 objects that satisfy our selection criteria and have more than one epoch of spectroscopy, we find 10 examples of quasars that have variable and/or ‘changing-look’ BEL features. Four of our objects have emerging BELs, five have disappearing BELs, and one object shows tentative evidence for having both emerging and disappearing BELs. With redshifts in the range 0.20 15 per cent of strongly variable luminous quasars display changing-look BEL features on rest-frame time-scales of 8 to 10 yr. Plausible time-scales for variable dust extinction are factors of 2–10 too long to explain the dimming and brightening in these sources, and simple dust reddening models cannot reproduce the BEL changes. On the other hand, an advancement such as disc reprocessing is needed if the observed variations are due to accretion rate changes.Publisher PDFPeer reviewe

The Malaria Secretome: From Algorithms to Essential Function in Blood Stage Infection

The malaria agent Plasmodium falciparum is predicted to export a “secretome” of several hundred proteins to remodel the host erythrocyte. Prediction of protein export is based on the presence of an ER-type signal sequence and a downstream Host-Targeting (HT) motif (which is similar to, but distinct from, the closely related Plasmodium Export Element [PEXEL]). Previous attempts to determine the entire secretome, using either the HT-motif or the PEXEL, have yielded large sets of proteins, which have not been comprehensively tested. We present here an expanded secretome that is optimized for both P. falciparum signal sequences and the HT-motif. From the most conservative of these three secretome predictions, we identify 11 proteins that are preserved across human- and rodent-infecting Plasmodium species. The conservation of these proteins likely indicates that they perform important functions in the interaction with and remodeling of the host erythrocyte important for all Plasmodium parasites. Using the piggyBac transposition system, we validate their export and find a positive prediction rate of ∼70%. Even for proteins identified by all secretomes, the positive prediction rate is not likely to exceed ∼75%. Attempted deletions of the genes encoding the conserved exported proteins were not successful, but additional functional analyses revealed the first conserved secretome function. This gave new insight into mechanisms for the assembly of the parasite-induced tubovesicular network needed for import of nutrients into the infected erythrocyte. Thus, genomic screens combined with functional assays provide unexpected and fundamental insights into host remodeling by this major human pathogen