The Symbiotic Anthozoan: A Physiological Chimera between Alga and Animal

The symbiotic life style involves mutual ecological, physiological,structural, and molecular adaptations between the partners.In the symbiotic association between anthozoans and photosyntheticdinoflagellates (Symbiodinium spp., also called zooxanthellae),the presence of the endosymbiont in the animal cells has constrainedthe host in several ways. It adopts behaviors that optimizephotosynthesis of the zooxanthellae. The animal partner hashad to evolve the ability to absorb and concentrate dissolvedinorganic carbon from seawater in order to supply the symbiont'sphotosynthesis. Exposing itself to sunlight to illuminate itssymbionts sufficiently also subjects the host to damaging solarultraviolet radiation. Protection against this is provided bybiochemical sunscreens, including mycosporine-like amino acids,themselves produced by the symbiont and translocated to thehost. Moreover, to protect itself against oxygen produced duringalgal photosynthesis, the cnidarian host has developed certainantioxidant defenses that are unique among animals. Finally,living in nutrient-poor waters, the animal partner has developedseveral mechanisms for nitrogen assimilation and conservationsuch as the ability to absorb inorganic nitrogen, highly unusualfor a metazoan. These facts suggest a parallel evolution ofsymbiotic cnidarians and plants, in which the animal host hasadopted characteristics usually associated with phototrophicorganisms.     

Portal sensing of intestinal gluconeogenesis is a mechanistic link in the diminution of food intake induced by diet protein

Protein feeding is known to decrease hunger and subsequent food intake in animals and humans. It has also been suggested that glucose appearance into portal vein, as occurring during meal assimilation, may induce comparable effects. Here, we connect these previous observations by reporting that intestinal gluconeogenesis (i.e., de novo synthesis of glucose) is induced during the postabsorptive time (following food digestion) in rats specifically fed on protein-enriched diet. This results in glucose release into portal blood, counterbalancing the lowering of glycemia resulting from intestinal glucose utilization. Comparable infusions into the portal vein of control postabsorptive rats (fed on starch-enriched diet) decrease food consumption and activate the hypothalamic nuclei regulating food intake. Similar hypothalamic activation occurs on protein feeding. All these effects are absent after denervation of the portal vein. Thus, portal sensing of intestinal gluconeogenesis may be a novel mechanism connecting the macronutrient composition of diet to food intake.     

Cooperation of amphiregulin and insulin-like growth factor-1 inhibits Bax- and Bad-mediated apoptosis via a protein kinase C-dependent pathway in non-small cell lung cancer cells

Amphiregulin (AR) and insulin-like growth factor-1 (IGF1) are growth factors known to promote non-small cell lung cancer (NSCLC) survival. We have previously published that 1) AR and IGF1, secreted by H358 NSCLC cells, cooperate to protect those cells and H322 NSCLC cells from serum-starved apoptosis; 2) H358 cells resist Bax-induced apoptosis through an inhibition of Bax conformational change. We show here that the antiapoptotic activity of the AR/IGF1 combination is specifically abolished by the PKC inhibitors calphostin C and staurosporine, but not by the MAPK and phosphatidylinositol 3-kinase inhibitors PD98059 and wortmannin, suggesting the involvement of a PKC-dependent and MAPK- and phosphatidylinositol 3-kinase-independent survival pathway. The PKCdelta inhibitor rottlerin restores apoptosis induced by serum deprivation. In addition, phosphorylation of PKCdelta and PKCzeta/lambda, but not of PKCalpha/beta(II), increases in serum-starved H358 cells and in H322 cells treated with an AR/IGF1 combination and is blocked by calphostin C. The combination of AR and IGF1 increases p90(rsk) and Bad phosphorylation as well as inhibiting the conformational change of Bax by a PKC-dependent mechanism. Finally, PKCdelta, PKCzeta, or p90(rsk) small interfering RNAs block the antiapoptotic activity of AR/IGF1 combination but have no effect on partial apoptosis inhibition observed with each factor used alone. Constitutively active PKC expression inhibits serum deprivation-induced apoptosis, whereas a catalytically inactive form of p90(rsk) restores it. Thus, AR and IGF1 cooperate to prevent apoptosis by activating a specific PKC-p90(rsk)-dependent pathway, which leads to Bad and Bax inactivation. This signaling pathway is different to that used by single factor.     

Thaumatocotyle pseudodasybatis Hargis, 1955 (Monogenea: Monocotylidae) from Aetobatus cf. narinari, with a comparison of specimens from Australia, French Polynesia and New Caledonia

Thaumatocotyle pseudodasybatis Hargis, 1955, has previously been described from Aetobatus narinari in the Atlantic and subsequently recorded from the Pacific. Aetobatus cf. narinari is now considered a species complex; as monocotylids are often strictly species specific, we test the hypothesis that detailed examination of specimens of monocotylids from rays from various localities could reveal morphological differences and eventually help our understanding of the systematics of the host. T. pseudodasybatis, previously known from seven specimens only, is redescribed from an additional 26 specimens from the South Pacific (off New Caledonia, Australia and Ranguiroa, French Polynesia), all from Aetobatus cf. narinari. The female reproductive organs are described in detail. The distal extremity of the male sclerotised copulatory organ, described in detail for the first time, shows a characteristic pattern of longitudinal striations on its edge that might be useful for future distinction from other species. The development of the male and female organs in juveniles is described, showing that growth of the male sclerotised copulatory organ begins with its basal part and precedes development of the ejaculatory bulb. Specimens from New Caledonia, Australia and French Polynesia had similar measurements and morphology, especially in the shape of the distal end of the male sclerotised copulatory organ; they were also similar to the holotype from the Atlantic. This suggests that all specimens from the Pacific and Atlantic belong to a single species; T. pseudodasybatis thus cannot be used to differentiate populations of Aetobatus cf. narinari, perhaps because this monocotylid is not strictly species-specific.

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Résumé Thaumatocotyle pseudodasybatis Hargis, 1955 a été décrit d’Aetobatus narinari dans l’Atlantique, et plus tard retrouvé dans le Pacifique. Aetobatus cf. narinari est maintenant considéré comme un complexe d’espèces. Parce que les Monocotylidae sont souvent strictement spécifiques de leur espèce-hôte, nous testons l’hypothèse qu’un examen détaillé des spécimens de Monocotylidae de raies de localités variées pourrait révéler des différences morphologiques et finalement aider à comprendre la systématique de l’hôte. T. pseudodasybatis, connu précédemment par seulement sept spécimens, est redécrit à partir de 26 spécimens supplémentaires de l’Océan Pacifique Sud (Nouvelle-Calédonie, Australie, et Ranguiroa, Polynésie Française), tous de Aetobatus cf. narinari. Les organes reproducteurs femelles sont décrits en détail. L’extrémité distale de l’appareil copulateur sclérifié mâle, décrite en détail pour la première fois, montre un patron caractéristique de striations longitudinales sur son bord, qui pourrait être utile à l’avenir pour différencier cette espèce. Le développement des organes mâles et femelles chez les juvéniles est décrit; cela montre que la croissance de l’appareil copulateur sclérifié mâle commence par son extrémité basale et précède le développement du bulbe éjaculateur. Les spécimens de Nouvelle-Calédonie, Australie et Polynésie Française ont une morphologie et des mensurations similaires, en particulier pour la forme de l’extrémité distale de l’appareil copulateur sclérifié mâle. Ils sont aussi similaires à l’holotype, qui vient de l’Atlantique. Ceci suggère que tous les spécimens du Pacifique et de l’Atlantique appartiennent à une même espèce. T. pseudodasybatis ne peut donc pas être utilisé pour différencier les populations d’Aetobatus cf. narinari, peut-être parce que ce Monocotylidae n’est pas strictement spécifique.

     

Plasticity to soil water deficit in Arabidopsis thaliana: dissection of leaf development into underlying growth dynamic and cellular variables reveals invisible phenotypes

Genetic variability in the plasticity of leaf area expansion in response to water deficit has been reported in Arabidopsis thaliana. Here, the objective was to identify the underlying dynamic and cellular processes involved in this variability. Twenty-five accessions were subjected to identical soil water deficit treatments. In all accessions, the plasticity of leaf production was low compared with that of individual leaf expansion. A subset of accessions was selected for further dissection of individual leaf expansion into its underlying variables: the rate and duration of leaf expansion and epidermal cell number and area. In all accessions, water deficit had opposite effects on the rate and duration of leaf expansion. The accumulation of these effects was reflected in changes in final leaf area. At the cellular level, moderate water deficits had opposite effects on cell number and cell size, but more severe ones reduced both variables. The importance of these opposing effects is highlighted by the behaviour of the accession An-1, for which the compensation between the decrease in leaf expansion rate and the increase in the duration of expansion is total. This dynamic plasticity in response to water deficit is not detectable when only final measurements are done.     

Yeast cytochrome c oxidase: a model system to study mitochondrial forms of the haem-copper oxidase superfamily

The known subunits of yeast mitochondrial cytochrome c oxidase are reviewed. The structures of all eleven of its subunits are explored by building homology models based on the published structures of the homologous bovine subunits and similarities and differences are highlighted, particularly of the core functional subunit I. Yeast genetic techniques to enable introduction of mutations into the three core mitochondrially-encoded subunits are reviewed.     

New insights into the organisation and intracellular localisation of the two subunits of glucose-6-phosphatase

Glucose-6 phosphatase (G6Pase), a key enzyme of glucose homeostasis, catalyses the hydrolysis of glucose-6 phosphate (G6P) to glucose and inorganic phosphate. A deficiency in G6Pase activity causes type 1 glycogen storage disease (GSD-1), mainly characterised by hypoglycaemia. Genetic analyses of the two forms of this rare disease have shown that the G6Pase system consists of two proteins, a catalytic subunit (G6PC) responsible for GSD-1a, and a G6P translocase (G6PT), responsible for GSD-1b. However, since their identification, few investigations concerning their structural relationship have been made. In this study, we investigated the localisation and membrane organisation of the G6Pase complex. To this aim, we developed chimera proteins by adding a fluorescent protein to the C-terminal ends of both subunits. The G6PC and G6PT fluorescent chimeras were both addressed to perinuclear membranes as previously suggested, but also to vesicles throughout the cytoplasm. We demonstrated that both proteins strongly colocalised in perinuclear membranes. Then, we studied G6PT organisation in the membrane. We highlighted FRET between the labelled C and N termini of G6PT. The intramolecular FRET of this G6PT chimera was 27%. The coexpression of unlabelled G6PC did not modify this FRET intensity. Finally, the chimera constructs generated in this work enabled us for the first time to analyze the relationship between GSD-1 mutations and the intracellular localisation of both G6Pase subunits. We showed that GSD1 mutations did neither alter the G6PC or G6PT chimera localisation, nor the interaction between G6PT termini. In conclusion, our results provide novel information on the intracellular distribution and organisation of the G6Pase complex.     

Implication of Glutathione in the In Vitro Antiplasmodial Mechanism of Action of Ellagic Acid

The search for new antimalarial chemotherapy has become increasingly urgent due to parasite resistance to current drugs. Ellagic acid (EA) is a polyphenol, recently found in various plant products, that has effective antimalarial activity in vitro and in vivo without toxicity. To further understand the antimalarial mechanism of action of EA in vitro, we evaluated the effects of EA, ascorbic acid and N-acetyl-L-cysteine (NAC), alone and/or in combination on the production of reactive oxygen species (ROS) during the trophozoite and schizonte stages of the erythrocytic cycle of P. falciparum. The parasitized erythrocytes were pre-labelled with DCFDA (dichlorofluorescein diacetate). We showed that NAC had no effect on ROS production, contrary to ascorbic acid and EA, which considerably reduced ROS production. Surprisingly, EA reduced the production of the ROS with concentrations (6.6×10⁻⁹ − 6.6×10⁻⁶ M) ten-fold lower than ascorbic acid (113×10⁻⁶ M). Additionally, the in vitro drug sensitivity of EA with antioxidants showed that antiplasmodial activity is independent of the ROS production inside parasites, which was confirmed by the additive activity of EA and desferrioxamine. Finally, EA could act by reducing the glutathione content inside the Plasmodium parasite. This was consolidated by the decrease in the antiplasmodial efficacy of EA in the murine model Plasmodium yoelii- high GSH strain, known for its high glutathione content. Given its low toxicity and now known mechanism of action, EA appears as a promising antiplasmodial compound.