Search

International audience; According to the literature, biological processes in the rhizosphere could play a role in the adaptation of plants to droughts under a changing climate. A previous study has identified significantly different productivity level and fruit quality for two tomato varieties under water deficit conditions. We conducted a field study, with and without water deficit, with these two varieties to examine whether microbiological activity and exopolysaccharides concentration could affect soil hydrophysical properties. The rhizosphere soil had indeed distinct bio-chemical and hydrophysical properties between the two cultivars and between the two water-related conditions. The quantity of soil exopolysaccharide and/or nitrogenous substances, and the activity of microorganisms (fungi in particular) explains part of the soil water retention measurements. In addition, these mechanisms are significantly accentuated for the cultivar with the best productive capability under water-limited condition—i.e. with commercial yield, fruit dry matter and water use efficiency which are respectively 35%, 28%, and 31% higher for the productive cultivar.

The design of enzymatic routes for the production of biosourced copolymers represents an attractive alternative to chemical synthesis from fossil carbon. In this paper, we explore the potential of glycosynthesizing enzymes to produce novel block copolymers composed of various covalently-linked α-glucans with contrasting structures and physicochemical properties. To this end, various glucansucrases able to synthesize α-glucans with different types of α-osidic bonds from sucrose were tested for their ability to elongate oligosaccharide and polysaccharide acceptors with different structures from the native polymer synthesized by each enzyme. We showed that two enzymes – namely, the alternansucrase from Leuconostoc mesenteroides NRRL B-1355 (specific for α(1 → 6)/α(1 → 3)-linked alternan synthesis) and the dextransucrase DSR-MΔ1 from Leuconostoc citreum NRRL B-1299 (specific for α(1 → 6)-linked dextran formation) – were able to elongate α(1 → 4)-linked amylose and α(1 → 6)/α(1 → 3)-linked alternan respectively. Carrying out stepwise acceptor reactions, and after optimization of the acceptor size and donor/acceptor ratio, two types of diblock copolymers were synthesized – a dextran-b-alternan and an alternan-b-amylose – as well as the triblock copolymer dextran-b-alternan-b-amylose. Their structural characterization, performed by combining chromatographic, NMR and permethylation analyses, showed that the copolymer polymerization degree ranged from 29 to 170, which is the highest degree of polymerization ever reported for an enzymatically synthesized polysaccharide-based copolymer. The addition of dextran and alternan blocks to amylose resulted in conformational modifications and related flexibility changes, as demonstrated by small angle X-ray scattering.

In French Atlantic coastal ponds of Charente, oysters can grow under conditions where phytoplankton production is limited by nutrients exhaustion. Such ponds typically show a high concentration of ciliates and flagellates during the growing season (1 x 104 to 3 x 105 cells l-1 in June 1997). In order to evaluate the importance of the "protozoan trophic link " for energy transfer from the " microbial food web" to large benthic suspension feeders, we offered a coastal pond community of ciliates and flagellates as potential prey to the oyster, Crassostrea gigas. Clearance rate, filtered particles and relative retention efficiency were evaluated. In the grazing experiment, 94 % of ciliates and 86 % of flagellates (size between 4 and 72 μm), were retained by the oyster. Whatever their size, protists were similarly retained by the oyster gills. In terms of carbon, oyster retain on average 126 μg carbon (C) h-1 g-1 dry weight, a value over 4 times higher than reported for phytoplankton. These results indicate that a field community of protists can contribute in coastal oyster rearing ponds to the energy requirements of the oyster Crassostrea gigas. We report here the first experimental evidence of a significant retention of a protist community by oysters, supporting the role of protists as a trophic link between picoplankton and benthic filter-feeding bivalves.

The linked concepts of 'microbial loop' and 'protozoan trophic link' have been very well documented in filter-feeding microzooplankton such as copepods, but have not been applied to energy transfer to benthic suspension-feeding macrofauna, with the exception of the recent demonstration of heterotrophic flagellate assimilation by mussels. The oyster Crassostrea gigas obtains energy resources by filtering microalgae (similar to 5 to 100 mu m). However, in turbid estuaries, light-limited phytoplanktonic production cannot entirely account for oyster energy requirements. Conversely, picoplankters (

Féral C, Le Gall S, Leymarie P. Angiotensin II modulates steroidogenesis in granulosa and theca in the rabbit ovary: its possible involvement in atresia. Eur J Endocrinol 1995;133:747–53. ISSN 0804–4643 Accumulating evidence has shown the ovary of mammals to contain an intrinsic renin–angiotensin system that has been ascribed an autocrine-paracrine role. The present study in the female rabbit ovary investigated the putative in vitro action of angiotensin II (A II) on basal and gonadotropin-induced steroidogenesis. Ovarian follicles from immature female rabbits treated with pregnant mare's serum gonadotropin (PMSG) were dissected out and a complete separation of the theca interna from the granulosa layer was performed, to demonstrate that A II affects separately the two individual cellular components of the follicular wall. We could show that theca is a source of estradiol whose production under human chorionic gonadotropin (hCG) stimulation was reduced by A II. At the same time, A II increased the in vitro hCG-stimulated secretion of testosterone by theca. In granulosa, A II decreased hCG-stimulated aromatization of androstenedione to estradiol but did not alter the release of hCG-stimulated progesterone production. These results suggest that A II could induce locally an increase in follicular fluid androgen/estrogen ratio and possibly participate in causing atresia. C. Féral, EP CNRS 009, Laboratoire de Biochimie, CHU Caen, F-14032 Caen Cedex, France

Background HIV-specific CD8 T cells responses rely on the recognition of peptide-MHC-I complexes by cognate T cell receptors. HIV-derived MHC-I epitopes result from the degradation of viral proteins by the cellular processing machinery including proteasomes and aminopeptidases. Interferon gamma changes proteasome composition and peptidase activities. We hypothesize that HIV infection might affect the expression or activities of the antigen processing machinery, either through a direct effect of the virus or indirectly through cellular activation or from the release of cytokines by surrounding infected cells.

Background Whether HIV-infectable subsets, such as CD4 T cells, monocytes, macrophages and dendritic cells (DCs), have equivalent capacity to produce and present MHC-I restricted epitopes to HIV-specific CD8 T cells is unknown. MHC-I epitopes are processed by an intracellular degradation pathway involving multiple proteases. In this study we analyzed the effect of toll-like receptor (TLR) agonist-mediated maturation on the processing and presentation of HIV antigens in monocyte-derived DCs and macrophages.

International audience; Lichens are complex organisms resulting from the symbiosis between fungus, microalga and/or cyanobacteria and are source of metabolites of interest. As other living organisms harboring bacterial communities they could be considered as a mini-ecosystem. These bacterial communities most often belong to different phyla: Actinobacteria, Proteobacteria, with a dominance of Alphaproteobacteria and Firmicutes. In this study, we focused on the bacterial communities present on six lichens from Brittany coast (France) (Roccella fuciformis, R. phycopsis, Lichina confinis, L. pygmaea, Xanthoria aureola and X. calcicola). Abundance and diversity of these communities are dependent on several extrinsic factors (environmental) and/or intrinsic parameters including the chemical composition of their substrates (lichens). So, our aims are to elucidate the chemical composition of the studied lichens (extraction, isolation and structural identification) as well as those of associated bacterial communities. Some bacterial species were isolated from these lichens, identified by molecular fingerprints and their culture were optimized (media composition, pH and temperature). Due to the existence of chemical interactions between symbionts we target molecules with antibiotic properties.

The circulating reninangiotensin system (RAS) participates in the regulation of blood pressure and electrolyte metabolism. Renin, a proteolytic enzyme, synthesized in the kidney from its biological precursor, prorenin, cleaves its substrate angiotensinogen in the blood to form the active octapeptide, angiotensin II (AII). All the RAS components are present in the reproductive system of mammals. During pregnancy, the level of prorenin increases in the plasma. The ovary is the source of this prorenin during early pregnancy and maternal decidua later on. During the menstrual cycle, the thecal of preovulatory follicles synthesize prorenin, renin and AII. Thecal renin synthesis is controlled by LH/hCG as demonstrated in vivo and in vitro in the rabbit. Ovarian renin seems to be identical to kidney renin. Prorenin appears to be the major secretory product rather than renin, which remains intracellular. AT2-type angiotensin II-receptors are expressed in the rat on follicular granulosa cells and could be down-regulated by FSH. The bovine thecal cells also express AT2-receptors, up-regulated by LH. These data are consistent with an autrocrine or paracrine role for ovarian RAS. It has been implicated in neovascularization of the follicle and regulation of steroidogenesis by increasing the androgen/estrogen ratio, an index of follicular atresia.

Hypermutation is an important mechanism used by different Salmonella enterica subspecies enterica to regulate genetic stability in adaptation to changing environments, including antimicrobial treatments and industrial processes. Strong hypermutator strains generally contain a mutation in genes of the methyl mismatch repair (MMR) system and have mutation frequencies up to 1000-fold higher than wild type strains. The objectives of this study were to determine the distribution of mutation frequencies from a collection of 209 Salmonella strains, to genetically characterize a strong mutator, and to study MMR mutated protein-DNA binding interactions. Only one strain of S. Heidelberg was determined to have a hypermutator phenotype by virtue of its high mutation rate. Sequencing of genes of the MMR system showed a 12bp deletion in the mutS gene was present. The MMR mutated protein-DNA binding interactions were studied by bioanalysis, using the available crystal structure of a similar MutS protein from Escherichia coli. This analysis showed the small deletion in the Salmonella MutS was localized within the core domain. A retardation assay with MutS from hypermutable and wild type strains showed this mutation has no effect on MutS DNA binding. A better understanding of the genetic mechanisms of hypermutation will help to anticipate the behavior of hypermutator strains in various conditions.

The origin of ovarian renin and its regulation by hCG were investigated in rabbit periovulatory follicles and cultured preovulatory follicular cells. Intracellular content of renin in thecal cells was 8-fold greater than of granulosa cells. In vivo, administration of hCG increased intracellular content of renin in thecal but not granulosa cells. Similar results were obtained for cultured follicular cells, from which renin was partly released into the medium. In vitro, hCG increased intracellular renin content of thecal but not granulosa cells, without obvious effect on release. Approximately 95% of ovarian renin was inactive, but could be activated by trypsin. Thecal renin was antagonized in vitro by renin antiserum, indicating a specific renin activity. Our study establishes in the rabbit the thecal cell origin of ovarian inactive renin and demonstrates hCG regulation of its synthesis.

International audience; The structure and summertime production of planktonic communities and the role of nondiatom planktonic cells were studied in coastal ponds, which are areas traditionally used for fattening and greening table-sized oysters. The abundance and biomass of nano–microplanktonic protists were determined at weekly intervals between February 1998 and February 1999 in a coastal pond without oysters in the French Atlantic coast near La Rochelle. The production of these microbiotas was determined in the summer period. The structure of plankton communities revealed the following observations: (1) microphytoplanktonic cells were mostly diatoms and dinoflagellates, (2) microzooplank-tonic cells were mainly ciliates, and (3) nanoplanktonic cells were represented by pigmented (80–90% of the nanoplankton biomass) and colorless nanoflagellates. Diatoms were dominated by Naviculiineae. Dinoflagellates were dominated by Peridiniales. Oligotrichida were predominant in the ciliate community. Protist biomass levels were nine times higher from April to August (summer period 1033 mg C L _ 1) than from September to March (winter period 114 mg C L _ 1). Whatever the season, nanoflagellates were dominant in the water column (66 and 53% of the entire protist biomass in the summer and winter periods, respectively). Nanoflagellates represented the highest production of nano–microplanktonic communities (76% of carbon protist production) in the coastal pond in summer and showed the shortest generation time (7.1 h). Dinoflagellates came after nanoflagellates in production (19.5% of carbon protist production). Diatoms represented only a supplementary carbon resource available for higher trophic levels, whereas, until now, they were considered as the principal food of oysters in coastal ponds. Ciliates were a small source of carbon, but their growth rate was high. We suggest, first, that nanoflagellates represented the primary resource available in the pond and could constitute an important food resource for higher trophic levels, such as oysters, farmed in this type of pond. Overall, the system appeared to be more autotrophic than hetero-trophic. Because inorganic nutrients are quickly exhausted in a semiclosed pond, pigmented flagellates dominated the carbon biomass, production and biomass of bacteria were high (thus, the microbial food web appeared to be active in this pond), and mixotrophy seemed to be an important trophic mode there.

The Nef protein from the human immunodeficiency virus (HIV) induces CD4 cell surface downregulation by interfering with the endocytic machinery. It has been recently proposed that binding of HIV type 1 Nef to the β subunit of COPI coatomers participated in the Nef-induced CD4 downregulation through recognition of a novel diacidic motif found in the C-terminal disordered loop of Nef (V. Piguet, F. Gu, M. Foti, N. Demaurex, J. Gruenberg, J. L. Carpentier, and D. Trono, Cell 97:63–73, 1999). We have mutated the glutamate residues which formed this motif in order to document this observation. Surprisingly, mutation of the diacidic sequence of Nef did not significantly affect its ability (i) to interact with β-COP, (ii) to downregulate CD4 cell surface expression, and (iii) to address an integral resident membrane protein containing Nef as the cytoplasmic domain to the endocytic pathway. Our results indicate that these acidic residues are not involved in the connection of Nef with the endocytic machinery through binding to β-COP. Additional studies are thus required to characterize the residues of Nef involved in the binding to β-COP and to evaluate the contribution of this interaction to the Nef-induced perturbations of membrane trafficking.

Background Epitopes displayed by MHC-I come from the multistep degradation of proteins by intracellular peptidases such as proteasome and aminopeptidases or cathepsins in the exogenous pathway. We hypothesize that due to structural homologies HIV protease inhibitors (PIs) used in antiretroviral therapies may affect activities of cellular peptidases involved in epitope processing and may affect epitope presentation to immune cells. Methods Using a fluorogenic assay the effect of 5 HIV-1 PIs (Ritonavir, Saquinavir, Nelfinavir, Indinavir, Atazanavir) on proteasome, aminopeptidase and cathepsin activities was tested in PBMCs from at least 6 healthy donors. Using PBMC cytosol as a source of peptidases and HPLC and mass spectrometry to define and quantify the degradation products, the effect of HIV PIs on HIV peptide processing kinetics and HIV epitope half-life was assessed. Finally we assessed the impact of PIs on the endogenous processing and presentation of epitopes by infected cells to CD8 T cells using a fluorescence-based cytotoxicity assay. Results HIV PIs variably altered proteasome, post-proteasomal aminopeptidases and cathepsin activities. Depending on the PI, some activities were inhibited (from 1.1 to 5 folds, p

Background HIV-1 Nef is a multifunctional protein frequently targeted by host CD8+ T cell responses in early and chronic phases of HIV-1 infection. In vivo reversions of CTL escape mutations within Nef have been reported, suggesting a possible impact of immune-selected mutations in Nef on viral fitness. The goal of this work was to determine whether CD8+ T cell selected mutations in regions outside of Gag, such as in Nef, also impair viral replication and may thus contribute to early immune control of HIV-1.

Les virus sont des ennemis des systemes immunitaires dans une guerre tres ancienne faite d'adaptations constantes aux armes et aux strategies de l'adversaire. Pour echapper a la detection et a la destruction par leur hote, les virus ont mis en place des mecanismes varies. Leur comprehension accroit nos connaissances sur les fonctionnements des systemes immunitaires et des virus. Les strategies mises en oeuvre par le virus de l'immunodeficience humaine (VIH) concernent notamment un processus-cle de la reponse immunitaire qui est la presentation antigenique par les molecules du complexe majeur d'histocompatibilite (CMH) aux lymphocytes T. Les cellules dendritiques sont des cellules specialisees dans la presentation antigenique possedant la propriete de mettre en route des reponses immunitaires. Dans l'infection par le VIH, les cellules dendritiques sont essentielles au virus pour sa penetration dans l'organisme, pour sa production et sa dissemination et, probablement, dans la pathogenie du SIDA.

In the Calyptraeidae, which are protandric hermaphrodite molluscs, the external male genital tract and the gonad develop synchronously. The penis appears in young animals but regresses at the time of sex inversion. Experiments carried out in Crepidula fornicata, using the organ culture technique, enabled us to prove that involution of the penis in vitro is not the result of an intrinsic cycle of this organ. Hormonal exposure which had occurred in vivo was sufficient to condition further development of the isolated penis in vitro. A pedal morphogenetic factor and a pleural regressive factor are responsible for the differentiation of the male genital tract. The programming depends upon a balance of these neuroendocrine factors in the animal that provided the explant. Notwithstanding its regression in vitro, the penis of Crepidula represents an organ unable to evolve by itself, inasmuch as its development is directly dependent on the neuroendocrine system.

The innervation of the endocrine optic gland of Sepia, which controls sexual maturation, was studied by immunocytochemistry. Anti-FMRFamide (Phe-Met-Arg-Phe-NH2) serum revealed immunoreactive neurons in the olfactory and basal-dorsal lobe of the supra-esophageal brain mass. The axons of these neurons form a network from which fibers run to the optic gland. The fibers form many varicosities on the glandular cells, indicating synaptic innervation. Apparently, the two brain lobes containing the immunopositive cells function as a unit where visual and olfactory cues are integrated to regulate the endocrine activity of the optic gland.

Very few investigations have been carried out about factors responsible for sex differentiation in protandric hermaphrodite molluscs. In Cvepidula plana, Gould (1947) showed the occurrence of a feminizing influence when individuals are isolated and a masculinizing effect of the female at the bottom of the chain on upper individuals of the association. However, experiments of Coe (1942) on Crepidula onyx and Wilczynsky (1958) on Crepidula fornicata, partly disprove Gould’s work.

International audience; The Mediterranean Thau Lagoon is an important European oyster farming area. Oyster growth levels are among the highest in France, although chlorophyll a concentration is low. Previous studies have demonstrated that picophytoplankton, nano-microphytoplankton, dinoflagellates and loricate ciliates such as tintinnids are abundant in the Thau Lagoon. Moreover, heterotrophic flagellates and aloricate ciliates have not been investigated. The aim of this study was to assess picophytoplankton, protist and zooplankton abundances in water columns of the Thau Lagoon and to understand the particular structure of the Thau microbial food web, which may explain such a paradoxical oyster growth. In oligotrophic waters in the Thau Lagoon, the picoeukaryote Ostreococcus tauri is the dominant autotrophic picoplankter with a maximum Summer abundance. On 17 August 1998, the picophytoplankton and nanophytoplankton abundances were not as high as expected and we observed the development of large diatoms. At this time, available carbon resources arose from microphytoplankton (84.5 %) and picoplanktonic cells represented only 1.27 % in terms of carbon. The heterotrophic cells were few in abundance and constitued only < 14 % of carbon resources. In order to evaluate the importance of the "protozoan trophic link" for energy transfer from "microbial food web" to large benthic suspension feeders, the oyster Crassostrea gigas was offered a planktonic community as potential prey. In the grazing experiment, all > 5 μm flagellates, microphytoplankton, dinoflagellates, ciliates and large zooplankton were retained by the oyster gills. Only < 5 μm flagellates and picoeukaryotic cells, Ostreococcus tauri, were not very well retained (45 % and 2 %). The high clearance rates of Crassostrea gigas found in this experiment can be explained by a low suspended particulate matter (0.65 mg l-1). Oysters adapted their retention mechanism when they lived in oligotrophic waters. These results indicate that, under the given experimental conditions, picophytoplankton did not represent a valuable trophic resource for farmed oysters because (1) Crassostrea can not retain picoparticles and (2) the picoplankton represented a poor available carbon resource to be transferred via a weak heterotrophic protist community. In the oyster pends of the Thau Lagoon during this study, which followed a rainfall event, microphytoplanktonic primary producers, in particular diatoms, could be considered as the main food sources for bivalve suspension feeders.

Macrophages in the healthy intestine are highly specialized and usually respond to the gut microbiota without provoking an inflammatory response. A breakdown in this tolerance leads to inflammatory bowel disease (IBD), but the mechanisms by which intestinal macrophages normally become conditioned to promote microbial tolerance are unclear. Strong epidemiological evidence linking disruption of the gut microbiota by antibiotic use early in life to IBD indicates an important role for the gut microbiota in modulating intestinal immunity. Here, we show that antibiotic use causes intestinal macrophages to become hyperresponsive to bacterial stimulation, producing excess inflammatory cytokines. Re-exposure of antibiotic-treated mice to conventional microbiota induced a long-term, macrophage-dependent increase in inflammatory T helper 1 (TH1) responses in the colon and sustained dysbiosis. The consequences of this dysregulated macrophage activity for T cell function were demonstrated by increased susceptibility to infections requiring TH17 and TH2 responses for clearance (bacterial Citrobacter rodentium and helminth Trichuris muris infections), corresponding with increased inflammation. Short-chain fatty acids (SCFAs) were depleted during antibiotic administration; supplementation of antibiotics with the SCFA butyrate restored the characteristic hyporesponsiveness of intestinal macrophages and prevented T cell dysfunction. Butyrate altered the metabolic behavior of macrophages to increase oxidative phosphorylation and also promoted alternative macrophage activation. In summary, the gut microbiota is essential to maintain macrophage-dependent intestinal immune homeostasis, mediated by SCFA-dependent pathways. Oral antibiotics disrupt this process to promote sustained T cell-mediated dysfunction and increased susceptibility to infections, highlighting important implications of repeated broad-spectrum antibiotic use.

Recently, it has been demonstrated that the human immunodeficiency virus type 1 (HIV-1) Nef from laboratory strains down-modulates cell surface expression of mature major histocompatibility complex class II (MHC-II) molecules, while up-regulating surface expression of the invariant chain (Ii) associated with immature MHC-II (P. Stumptner-Cuvelette, S. Morchoisne, M. Dugast, S. Le Gall, G. Raposo, O. Schwartz, and P. Benaroch, Proc. Natl. Acad. Sci. USA 98: 12144-12149, 2001). These Nef functions could contribute to impaired CD4 + -T-helper-cell responses found in HIV-1-infected patients with progressive disease. However, it is currently unknown whether nef alleles derived from HIV-1-infected individuals or from other primate lentiviruses also modulate MHC-II and Ii. In the present study, we demonstrate that both activities are conserved among primary HIV-1 nef alleles, as well as among HIV-2 and simian immunodeficiency virus (SIV) nef alleles. Down-modulation of mature MHC-II required high levels of Nef expression. In contrast, surface expression of Ii was already strongly increased at low to medium levels of Nef expression. Notably, nef genes derived from two of four HIV-1-infected long-term nonprogressors did not up-regulate Ii, whereas nef alleles derived from 10 individuals with progressive disease were active in this assay. Unlike other in vitro Nef functions, the average activity of Nef in modulating MHC-II and Ii surface expression did not change significantly during the course of infection. Mutational analysis confirmed that MHC-II down- and Ii up-regulation are functionally separable from each other and from other Nef functions and identified acidic residues, located at the base of the flexible C-proximal loop of Nef, that are critical for increased Ii expression. Overall, our results suggest that the ability of Nef to interfere with MHC-II antigen presentation might play a role in AIDS pathogenesis.