51 research outputs found

    Variation in mycorrhizal growth response among a spring wheat mapping population shows potential to breed for symbiotic benefit.

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    Funder: N8 Agrifood SchemeAll cereal crops engage in arbuscular mycorrhizal symbioses which can have profound, but sometimes deleterious, effects on plant nutrient acquisition and growth. The mechanisms underlying variable mycorrhizal responsiveness in cereals are not well characterised or understood. Adapting crops to realise mycorrhizal benefits could reduce fertiliser requirements and improve crop nutrition where fertiliser is unavailable. We conducted a phenotype screen in wheat (Triticum aestivum L.), using 99 lines of an Avalon × Cadenza doubled-haploid mapping population. Plants were grown with or without a mixed inoculum containing 5 species of arbuscular mycorrhizal fungi. Plant growth, nutrition and mycorrhizal colonisation were quantified. Plant growth response to inoculation was remarkably varied among lines, ranging from more than 30% decrease to 80% increase in shoot biomass. Mycorrhizal plants did not suffer decreasing shoot phosphorus concentration with increasing biomass as observed in their non-mycorrhizal counterparts. The extent to which mycorrhizal inoculation was beneficial for individual lines was negatively correlated with shoot biomass in the non-mycorrhizal state but was not correlated with the extent of mycorrhizal colonisation of roots. Highly variable mycorrhizal responsiveness among closely related wheat lines and the identification of several QTL for these traits suggests the potential to breed for improved crop-mycorrhizal symbiosis

    Knockout studies reveal an important role of <i>plasmodium</i> lipoic acid protein ligase a1 for asexual blood stage parasite survival

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    Lipoic acid (LA) is a dithiol-containing cofactor that is essential for the function of a-keto acid dehydrogenase complexes. LA acts as a reversible acyl group acceptor and 'swinging arm' during acyl-coenzyme A formation. The cofactor is post-translationally attached to the acyl-transferase subunits of the multienzyme complexes through the action of octanoyl (lipoyl): &lt;i&gt;N&lt;/i&gt;-octanoyl (lipoyl) transferase (LipB) or lipoic acid protein ligases (LplA). Remarkably, apicomplexan parasites possess LA biosynthesis as well as scavenging pathways and the two pathways are distributed between mitochondrion and a vestigial organelle, the apicoplast. The apicoplast-specific LipB is dispensable for parasite growth due to functional redundancy of the parasite's lipoic acid/octanoic acid ligases/transferases. In this study, we show that &lt;i&gt;LplA1&lt;/i&gt; plays a pivotal role during the development of the erythrocytic stages of the malaria parasite. Gene disruptions in the human malaria parasite &lt;i&gt;P.falciparum&lt;/i&gt; consistently were unsuccessful while in the rodent malaria model parasite &lt;i&gt;P. berghei&lt;/i&gt; the &lt;i&gt;LplA1&lt;/i&gt; gene locus was targeted by knock-in and knockout constructs. However, the &lt;i&gt;LplA1&lt;/i&gt; &lt;sup&gt;(-)&lt;/sup&gt; mutant could not be cloned suggesting a critical role of LplA1 for asexual parasite growth &lt;i&gt;in vitro&lt;/i&gt; and &lt;i&gt;in vivo&lt;/i&gt;. These experimental genetics data suggest that lipoylation during expansion in red blood cells largely occurs through salvage from the host erythrocytes and subsequent ligation of LA to the target proteins of the malaria parasite

    Access to primary care for socio-economically disadvantaged older people in rural areas: exploring realist theory using structural equation modelling in a linked dataset

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    Background: Realist approaches seek to answer questions such as ‘how?’, ‘why?’, ‘for whom?’, ‘in what circumstances?’ and ‘to what extent?’ interventions ‘work’ using context-mechanism-outcome (CMO) configurations. Quantitative methods are not well-established in realist approaches, but structural equation modelling (SEM) may be useful to explore CMO configurations. Our aim was to assess the feasibility and appropriateness of SEM to explore CMO configurations and, if appropriate, make recommendations based on our access to primary care research. Our specific objectives were to map variables from two large population datasets to CMO configurations from our realist review looking at access to primary care, generate latent variables where needed, and use SEM to quantitatively test the CMO configurations. Methods: A linked dataset was created by merging individual patient data from the English Longitudinal Study of Ageing and practice data from the GP Patient Survey. Patients registered in rural practices and who were in the highest deprivation tertile were included. Three latent variables were defined using confirmatory factor analysis. SEM was used to explore the nine full CMOs. All models were estimated using robust maximum likelihoods and accounted for clustering at practice level. Ordinal variables were treated as continuous to ensure convergence. Results: We successfully explored our CMO configurations, but analysis was limited because of data availability. Two hundred seventy-six participants were included. We found a statistically significant direct (context to outcome) or indirect effect (context to outcome via mechanism) for two of nine CMOs. The strongest association was between ‘ease of getting through to the surgery’ and ‘being able to get an appointment’ with an indirect mediated effect through convenience (proportion of the indirect effect of the total was 21%). Healthcare experience was not directly associated with getting an appointment, but there was a statistically significant indirect effect through convenience (53% mediated effect). Model fit indices showed adequate fit. Conclusions: SEM allowed quantification of CMO configurations and could complement other qualitative and quantitative techniques in realist evaluations to support inferences about strengths of relationships. Future research exploring CMO configurations with SEM should aim to collect, preferably continuous, primary data

    The Amidase Domain of Lipoamidase Specifically Inactivates Lipoylated Proteins In Vivo

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    BACKGROUND:In the 1950s, Reed and coworkers discovered an enzyme activity in Streptococcus faecalis (Enterococcus faecalis) extracts that inactivated the Escherichia. coli and E. faecalis pyruvate dehydrogenase complexes through cleavage of the lipoamide bond. The enzyme that caused this lipoamidase activity remained unidentified until Jiang and Cronan discovered the gene encoding lipoamidase (Lpa) through the screening of an expression library. Subsequent cloning and characterization of the recombinant enzyme revealed that lipoamidase is an 80 kDa protein composed of an amidase domain containing a classic Ser-Ser-Lys catalytic triad and a carboxy-terminal domain of unknown function. Here, we show that the amidase domain can be used as an in vivo probe which specifically inactivates lipoylated enzymes. METHODOLOGY/PRINCIPAL FINDINGS:We evaluated whether Lpa could function as an inducible probe of alpha-ketoacid dehydrogenase inactivation using E. coli as a model system. Lpa expression resulted in cleavage of lipoic acid from the three lipoylated proteins expressed in E. coli, but did not result in cleavage of biotin from the sole biotinylated protein, the biotin carboxyl carrier protein. When expressed in lipoylation deficient E. coli, Lpa is not toxic, indicating that Lpa does not interfere with any other critical metabolic pathways. When truncated to the amidase domain, Lpa retained lipoamidase activity without acquiring biotinidase activity, indicating that the carboxy-terminal domain is not essential for substrate recognition or function. Substitution of any of the three catalytic triad amino acids with alanine produced inactive Lpa proteins. CONCLUSIONS/SIGNIFICANCE:The enzyme lipoamidase is active against a broad range of lipoylated proteins in vivo, but does not affect the growth of lipoylation deficient E. coli. Lpa can be truncated to 60% of its original size with only a partial loss of activity, resulting in a smaller probe that can be used to study the effects of alpha-ketoacid dehydrogenase inactivation in vivo

    TRAF6 and IRF7 Control HIV Replication in Macrophages

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    The innate immune system recognizes virus infection and evokes antiviral responses which include producing type I interferons (IFNs). The induction of IFN provides a crucial mechanism of antiviral defense by upregulating interferon-stimulated genes (ISGs) that restrict viral replication. ISGs inhibit the replication of many viruses by acting at different steps of their viral cycle. Specifically, IFN treatment prior to in vitro human immunodeficiency virus (HIV) infection stops or significantly delays HIV-1 production indicating that potent inhibitory factors are generated. We report that HIV-1 infection of primary human macrophages decreases tumor necrosis factor receptor-associated factor 6 (TRAF6) and virus-induced signaling adaptor (VISA) expression, which are both components of the IFN signaling pathway controlling viral replication. Knocking down the expression of TRAF6 in macrophages increased HIV-1 replication and augmented the expression of IRF7 but not IRF3. Suppressing VISA had no impact on viral replication. Overexpression of IRF7 resulted in enhanced viral replication while knocking down IRF7 expression in macrophages significantly reduced viral output. These findings are the first demonstration that TRAF6 can regulate HIV-1 production and furthermore that expression of IRF7 promotes HIV-1 replication

    Présentation d'éclats de basalte trouvés, avec des os, du charbon, de très rares silex, et des poteries, dans une tranchée de la Roche-Sellée, près Murat

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    Pagès-Allary J. Présentation d'éclats de basalte trouvés, avec des os, du charbon, de très rares silex, et des poteries, dans une tranchée de la Roche-Sellée, près Murat. In: Bulletin de la Société préhistorique de France, tome 6, n°6, 1909. pp. 310-311

    Les Fouilles de Las Tours en 1910

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    Pagès-Allary J. Les Fouilles de Las Tours en 1910. In: Bulletin de la Société préhistorique de France, tome 8, n°4, 1911. pp. 265-270

    Hypothèses tirées des Haches Préhistoriques

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    Pagès-Allary J. Hypothèses tirées des Haches Préhistoriques. In: Bulletin de la Société préhistorique de France, tome 8, n°10, 1911. pp. 627-636

    Les Haches préhistoriques en Cuivre, Bronze et Fer ne sont, bien souvent, que des Outils d'agriculture ou des Talons de Lances, quand ce ne sont pas des Tranchets de corroyeurs

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    Pagès-Allary J. Les Haches préhistoriques en Cuivre, Bronze et Fer ne sont, bien souvent, que des Outils d'agriculture ou des Talons de Lances, quand ce ne sont pas des Tranchets de corroyeurs. In: Bulletin de la Société préhistorique de France, tome 10, n°5, 1913. pp. 304-309
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