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

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

Terminologie des ressources éducatives libres et définitions retenues

Ressources éducatives libresOpen educational resourcesLe présent document contient des définitions, des explications et des prises de position de la fabriqueREL concernant la terminologie relative aux ressources éducatives libres (REL). Certains termes n’ayant pas encore de définition consensuelle apparente, les explications sont alors privilégiées, notamment pour présenter les principales conceptions alternatives et les énoncés retenus.Ressource éducative libre (REL) -- Origine -- Définition -- Les 5R -- Les différents types de REL -- Position de la fabriqueREL -- Distinguer une REL -- Distinction de ressource numérique -- Distinction de ressource gratuite -- Distinction d’œuvre utilisable librement -- La faisabilité technique de l’exercice des droits accordés pour une REL -- Position de la fabriqueREL sur les considérations techniques des REL -- MétaREL -- Définition -- Position de la fabriqueREL sur les métaREL -- Licence -- Creative Commons et ses licences -- Licences imposées comme condition de financement -- Avantages et inconvénients des licences CC BY, SA et NC pour les REL -- Position de la fabriqueREL sur les licences CC -- Domaine public -- Travaux cités

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Investing in Saving Lives: Designing Second-Stage Women’s Shelters on First Nation Reserves

Most Indigenous women in Canada (61%) experience intimate partner violence (IPV), which is significantly worse than the high rate of 44 percent for other women in Canada. Despite the great risk for IPV, only three unfunded second-stage shelters for more than 600 First Nation reserves exist in Canada to provide First Nation women and their children a safe home. Second-stage housing offers IPV survivors transitional homes for an extended period that provide safety and renewal after their initial emergency shelter stays. This article documents the need for safe, nurturing, and culturally appropriate second-stage shelters for Indigenous women and their families to heal and rebuild. The authors provide two second-stage prototype designs based on domestic environmental analysis and concepts of houselessness, home, and co-housing. We discuss how these designs are one step in an action plan to protect Indigenous women and stop the genocide of Indigenous Peoples by supporting cultural, economic, health, and social development. The literature review and design concepts form an agenda to have design goals for housing IPV survivors that answers the “Calls to Justice for Murdered and Missing Women” and expands this needed service to every reserve

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

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

The Amidase Domain of Lipoamidase Specifically Inactivates Lipoylated Proteins In Vivo

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