An interaction between L-prostaglandin D synthase and arrestin increases PGD2 production

L-type prostaglandin synthase (L-PGDS) produces PGD(2), a lipid mediator involved in neuromodulation and inflammation. Here, we show that L-PGDS and arrestin-3 (Arr3) interact directly and can be co-immunoprecipitated endogenously from MG-63 osteoblasts. Perinuclear L-PGDS/Arr3 co-localization is observed in PGD(2)-producing MG-63 cells and is induced by the addition of the L-PGDS substrate or co-expression of COX-2 in HEK293 cells. Inhibition of L-PGDS activity in MG-63 cells triggers redistribution of Arr3 and L-PGDS to the cytoplasm. Perinuclear localization of L-PGDS is detected in wild-type mouse embryonic fibroblasts (MEFs) but is more diffused in MEFs-arr-2(-/-)-arr-3(-/-). Arrestin-3 promotes PGD(2) production by L-PGDS in vitro. IL-1β-induced PGD(2) production is significantly lower in MEFs-arr-2(-/-)-arr-3(-/-) than in wild-type MEFs but can be rescued by expressing Arr2 or Arr3. A peptide corresponding to amino acids 86-100 of arrestin-3 derived from its L-PGDS binding domain stimulates L-PGDS-mediated PGD(2) production in vitro and in MG-63 cells. We report the first characterization of an interactor/modulator of a PGD(2) synthase and the identification of a new function for arrestin, which may open new opportunities for improving therapies for the treatment of inflammatory diseases.     

A Host YB-1 Ribonucleoprotein Complex Is Hijacked by Hepatitis C Virus for the Control of NS3-Dependent Particle Production

Hepatitis C virus (HCV) orchestrates the different stages of its life cycle in time and space through the sequential participation of HCV proteins and cellular machineries; hence, these represent tractable molecular host targets for HCV elimination by combination therapies. We recently identified multifunctional Y-box-binding protein 1 (YB-1 or YBX1) as an interacting partner of NS3/4A protein and HCV genomic RNA that negatively regulates the equilibrium between viral translation/replication and particle production. To identify novel host factors that regulate the production of infectious particles, we elucidated the YB-1 interactome in human hepatoma cells by a quantitative mass spectrometry approach. We identified 71 YB-1-associated proteins that included previously reported HCV regulators DDX3, heterogeneous nuclear RNP A1, and ILF2. Of the potential YB-1 interactors, 26 proteins significantly modulated HCV replication in a gene-silencing screening. Following extensive interaction and functional validation, we identified three YB-1 partners, C1QBP, LARP-1, and IGF2BP2, that redistribute to the surface of core-containing lipid droplets in HCV JFH-1-expressing cells, similarly to YB-1 and DDX6. Importantly, knockdown of these proteins stimulated the release and/or egress of HCV particles without affecting virus assembly, suggesting a functional YB-1 protein complex that negatively regulates virus production. Furthermore, a JFH-1 strain with the NS3 Q221L mutation, which promotes virus production, was less sensitive to this negative regulation, suggesting that this HCV-specific YB-1 protein complex modulates an NS3-dependent step in virus production. Overall, our data support a model in which HCV hijacks host cell machinery containing numerous RNA-binding proteins to control the equilibrium between viral RNA replication and NS3-dependent late steps in particle production.     

Integration of cell line and process development to overcome the challenge of a difficult to express protein

This case study addresses the difficulty in achieving high level expression and production of a small, very positively charged recombinant protein. The novel challenges with this protein include the protein's adherence to the cell surface and its inhibitory effects on Chinese hamster ovary (CHO) cell growth. To overcome these challenges, we utilized a multi‐prong approach. We identified dextran sulfate as a way to simultaneously extract the protein from the cell surface and boost cellular productivity. In addition, host cells were adapted to grow in the presence of this protein to improve growth and production characteristics. To achieve an increase in productivity, new cell lines from three different CHO host lines were created and evaluated in parallel with new process development workflows. Instead of a traditional screen of only four to six cell lines in bioreactors, over 130 cell lines were screened by utilization of 15 mL automated bioreactors (AMBR) in an optimal production process specifically developed for this protein. Using the automation, far less manual intervention is required than in traditional bench‐top bioreactors, and much more control is achieved than typical plate or shake flask based screens. By utilizing an integrated cell line and process development incorporating medium optimized for this protein, we were able to increase titer more than 10‐fold while obtaining desirable product quality. Finally, Monte Carlo simulations were performed to predict the optimal number of cell lines to screen in future cell line development work with the goal of systematically increasing titer through enhanced cell line screening. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1201–1211, 2015     

Caractérisation des peuplements de macroinvertébrés benthiques à l’aide de la carte auto-organisatrice (SOM)

Macrobenthic organisms are much used nowadays as bio-indicators for their ability to reflect different disturbances of aquatic ecosystems. However, they have never been used to assess the ecological status of the lagoons of Benin. The current study aims at revealing the ecological state of the lagoon of Porto-Novo through the study of changes in the settlement of benthic organisms collected in this lagoon. The sampling was carried out from July 2007 to June 2009 on a frequency of four seasons of collection per year. The self-organizing map (SOM) of Kohonen has been used for various patterns of distribution of collected organisms. A discriminant analysis (AFD) has allowed the identification of the parameters that govern the patterns observed in this environment. Four groups of macrobenthic communities emerged that were well predicted (75%). The distribution of benthic macro invertebrates of this lagoon is therefore zonal, seasonal and discriminated by variables of mineralization and sediment grain size. The difference in taxonomic richness corresponds to environmental conditions of stations appearing more or less stable and highlighting a gradient of the stress on organisms. Places with unstable conditions are affected by human activities due to their proximity to homes that are enriched in organic matter. These places are full of polluted-tolerant species such as the Gastropods Potamididae, Cirratulidae Polychaetes, Diptera Chironomidae and Oligochaeta. The macrobenthic fauna, which is well differentiated in the groups, is then subjected to human disturbance.     

Role of I-region gene products in T cell activation. I. Stimulation of T lymphocyte proliferative responses by subcellular membrane preparations containing Ia alloantigens

A model system has been developed for exploring the requirements for activation of T cells by subcellular forms of Ia alloantigen. Lymph node cells from mice recently primed subcutaneously with viable allogeneic cells show strong proliferative responses in vitro to membrane preparations derived from cells bearing the appropriate I-region-encoded glycoproteins. This stimulation shows kinetics characteristic of a secondary response, with a peak at 24 to 48 hr. Primary responses to alloantigen-bearing membranes are weak or absent under these conditions. The predominant cell type involved in the secondary response is the Lyt-1+ T lymphocyte, and the major antigenic stimulus is the I-A subregion-encoded Ia glycoprotein. Syngeneic Ia+ accessory cells do not appear necessary for activation to occur. Detergent solubilized reconstituted membrane vesicles also will stimulate primed T lymphocytes to respond by proliferation. The applications of this approach to the study of T cell recognition of antigen and the role of nonspecific lymphokines in T cell triggering are discussed.     

Influence des inflorescences de la plante hôte (Vigna unguiculata) sur la levée de la diapause reproductrice de Bruchidius atrolineatus

Bruchidius atrolineatus (Pic) présente en zone sahélienne une diapause reproductrice durant la saison sèche et une partie de la saison des pluies. Les femelles diapausantes ne produisent pas de vitellogénine et le germarium des ovarioles est seul développé. Chez les mâles la spermatogenèse est très ralentie et les glandes annexes sont inactives. Lorsque les insectes diapausants sont placés en présence d'inflorescences de Vigna unguiculata Walp, leurs organes reproducteurs deviennent fonctionnels après un temps de latence de 15 à 20 jours. Il n'y a par contre aucune levée de la diapause chez des bruches placées en présence de gousses sèches de V. unguiculata dans une atmosphère saturée. Des informations sensorielles issues des pièces florales ou (et) des facteurs nutritionnels sans doute liés a la consommation de nectar semblent être à l'origine de cette levée de la diapause. Le pollen fort peu consommé n'a par contre aucun rôle. Cette régulation du cycle reproducteur de B. atrolineatus par les inflorescences de V. unguiculata permet l'émission des oeufs dès que les gousses commencent à se former à la fin de la saison des pluies.

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Summary Bruchidius atrolineatus (Pic) is a widely distributed bruchid in the Sahelian zone which shows a reproductive diapause during the dry season and part of the rainy season. Diapausing females do not produce vitellogenin and their ovaries are reduced to the germarium. Spermatogenesis is very much reduced and male accessory glands are inactive. When these insects were placed in the presence of inflorescences of Vigna unguiculata which were renewed daily, the reproductive diapause of both males and females was interrupted after 15–20 days. Vitellogenesis occurred in the females and spermatogenesis increased in the males whilst their accessory glands became functional. When diapausing bruchids, found in stores of on V. unguiculata seeds during the dry season, were placed near the host plant's inflorescences, diapause was also terminated. In all cases, diapause was not interrupted when the insects were offered dry pods of V. unguiculata in a water-saturated atmosphere. The pollen, which is hardly eaten by this bruchid, did not seem to stimulate termination of diapause. Sensory stimulations derived from the flowers or/and nutritional factors may be the cause of the development of the reproductive organs. After termination of the diapause the males showed normal sexual activity whereas female fecundity was rather low, at least in our experimental conditions. This type of reproductive regulation allows this sahelian bruchid to resume its sexual activity when the host plant's flowers appear in the field at the end of the rainy season. Then the beetles lay their eggs on the pods as soon as the pods are developed.

     

Crop mixtures: does niche complementarity hold for belowground resources? An experimental test using rice genotypic pairs

Aims

A growing body of research supports the feasibility of biocrust rehabilitation. Identifying populations of key species that are amenable to cultivation and that are resilient in rehabilitation contexts would advance the efficacy of these technologies. Here we investigate the growth and stress response of the cosmopolitan biocrust moss, Syntrichia ruralis.

Methods

We sampled populations of S. ruralis along a precipitation seasonality gradient from the Colorado Plateau ecoregion of the western United States. We cultivated these populations in an experiment manipulating duration of hydration periods on a weekly cycle. We then treated greenhouse grown materials with brief, stressful watering events, measuring how many events they could survive.

Results

All populations grew at an accelerated rate compared to growth in a natural setting, at least doubling biomass in five months. Increasing duration of hydration periods led to more growth in all but one population. Volunteer biocrust algae and cyanobacteria developed during cultivation, and differed among populations. Greenhouse grown mosses differed in their response to stressful watering, with the most susceptible populations dying at half the number events compared to the most tolerant.

Conclusions

These findings argue for informed selection and deployment of Syntrichia ruralis populations for soil rehabilitation.

     

Animal-assisted Psychotherapy and Trauma: A Meta-analysis

The present meta-analysis examined the efficacy of animal-assisted psychotherapy for individuals who have experienced trauma. Eight studies quantitatively assessed the treatment effects of involvement in animal-assisted psychotherapy. A random effects model was used to aggregate each study into an overall effect size. Eight effect sizes were included in the pre-versus post-comparison analysis. The results indicate a large effect size (Hedge’s g = 0.86, p < 0.001, 95% CI [.53, 1.18]). Two effect sizes were included in the treatment versus control comparison analysis. The results indicated a small to moderate effect size (g = 0.46, p = 0.03, 95% CI [0.04, 0.06]). Limited moderator analyses were able to be conducted due to lack of consistent reporting across studies. Place of study and percentage of female participants in the treatment group were found to statistically moderate the effect of animal-assisted psychotherapy. The results indicate that animal-assisted therapy is an efficacious treatment for trauma.