Influence of culture conditions on recombinant Drosophila melanogaster S2 cells producing rabies virus glycoprotein cultivated in serum-free medium

The recombinant G glycoprotein from the surface of the rabies virus (RVGP) is a promising candidate as a rabies vaccine component and also for diagnostic purposes. In this study, RVGP production by transfected Drosophila melanogaster S2 cells cultivated in a serum-free medium (supplemented IPL-41 medium) was carried out. The effects of pH and pO₂ were evaluated in batch culture in parallel spinner flasks. The use of a pH equal to 6.3 and a pO₂ of 40% air saturation resulted in the highest RVGP content. These conditions were also used in fed-batch mode, yielding a RVGP content level of 98 g/10⁷ cells. The main nutrients consumed were glucose, glutamine, asparagine, serine and proline and the major metabolites produced were alanine and ammonia, according to the metabolism studies performed. Since RVGP is a transmembrane protein, two different methods for protein recovery were assessed and compared. Detergent-based cell disruption showed to be more effective than mechanical disruption with glass beads for glycoprotein recovery.     

Two sympatric Byrsonima species (Malpighiaceae) differ in phenological and reproductive patterns

Sympatric and taxonomically related species may present pre- and/or postzygotic mechanisms for reproductive isolation. We compared the phenology and reproductive biology of Byrsonima intermedia and B. pachyphylla, two sympatric Malpighiaceae species, growing in a remnant of Cerrado in Central Brazil. Both species are evergreens and do not have an overlapping flowering period. In both species, dispersal occurs during the rainy season with low and intermediate overlap in the production of immature and mature fruits, respectively. Both species exhibit self-incompatibility (SI), or, in the case of B. intermedia, facultative SI, and, hence, depend on pollinators to compensate for the lack of spontaneous self-pollination. The flowers of the Byrsonima species were visited by fifteen species of bees that collected pollen and/or oil. Based on their more appropriate intrafloral posture, Epicharis flava and Centris varia were the main pollinators of both species, presenting more frequent visits and/or exhibiting trapline foraging behavior, tending to promote outcrossing. Temporal variation in flowering period and total or partial self-incompatibility seem to function as prezygotic isolation mechanisms that form barriers to gene flow between the studied species and probably avoid competition among pollinators. Both species make oil and pollen available to flower visitors and pollinators almost the entire year, but since Byrsonima intermedia has a long flowering time and is very abundant in the study area, it seems to be more important in maintaining flower-visiting bees.     

Characterization of kefir yeasts with antifungal capacity against Aspergillus species

International Microbiology - Kefir is a fermented probiotic drink obtained by placing kefir granules in a suitable substrate. The kefir granules are a consortium of bacteria and yeasts embedded in...     

Microbial communities of the Mediterranean rocky shore: ecology and biotechnological potential of the sea-land transition

Microbial communities from harsh environments hold great promise as sources of biotechnologically relevant strains and compounds. In the present work, we have characterized the microorganisms from the supralittoral and splash zone in three different rocky locations of the Western Mediterranean coast, a tough environment characterized by high levels of irradiation and large temperature and salinity fluctuations. We have retrieved a complete view of the ecology and functional aspects of these communities and assessed the biotechnological potential of the cultivable microorganisms. All three locations displayed very similar taxonomic profiles, with the genus Rubrobacter and the families Xenococcaceae, Flammeovirgaceae, Phyllobacteriaceae, Rhodobacteraceae and Trueperaceae being the most abundant taxa; and Ascomycota and halotolerant archaea as members of the eukaryotic and archaeal community respectively. In parallel, the culture-dependent approach yielded a 100-isolates collection, out of which 12 displayed high antioxidant activities, as evidenced by two in vitro (hydrogen peroxide and DPPH) and confirmed in vivo with Caenorhabditis elegans assays, in which two isolates, CR22 and CR24, resulted in extended survival rates of the nematodes. This work is the first complete characterization of the Mediterranean splash-zone coastal microbiome, and our results indicate that this microbial niche is home of an extremophilic community that holds biotechnological potential.     

Salvage Logging Versus the Use of Burnt Wood as a Nurse Object to Promote Post‐Fire Tree Seedling Establishment

Intense debate surrounds the effects of post‐fire salvage logging (SL) versus nonintervention policies on forest regeneration, but scant support is available from experimental studies. We analyze the effect of three post‐fire management treatments on the recruitment of a serotinous pine (Pinus pinaster) at a Mediterranean mountain. Treatments were applied 7 months after the fire and differ in the degree of intervention, ranging from “no intervention” (NI, all trees left standing) to “partial cut plus lopping” (PCL, felling most of the trees, cutting the main branches, and leaving all the biomass in situ without mastication), and “SL” (felling and piling the logs, and masticating the woody debris). Seedling survival after 3 years was the highest in PCL (47.3% versus 38.7% in SL). This was associated with the amelioration of microclimatic conditions under the scattered branches, which reduced radiation and soil temperature while increasing soil moisture. Seedling density after 2 years was approximately 5.5 times higher in PCL than in SL, as in SL a large fraction of seedlings was lost as a consequence of mechanized mastication. The NI treatment showed the lowest seedling survival (17.3%). Nevertheless, seedling density was similar to SL. Seedling growth scarcely differed among treatments. Our results show that branches left onsite acted as nurse objects that improved key microclimatic conditions for seedling recruitment. This creates a facilitative interaction ideal for seedling establishment in moisture‐deficient ecosystems, as it provides the benefit of a shading overstory but without underground competition.     

Diadenosine tetraphosphate (Ap<Subscript>4</Subscript>A) inhibits ATP-induced excitotoxicity: a neuroprotective strategy for traumatic spinal cord injury treatment

Reducing cell death during the secondary injury is a major priority in the development of a cure for traumatic spinal cord injury (SCI). One of the earliest processes that follow SCI is the excitotoxicity resulting from the massive release of excitotoxicity mediators, including ATP, which induce an excessive and/or prolonged activation of their receptors and a deregulation of the calcium homeostasis. Diadenosine tetraphosphate (Ap₄A) is an endogenous purinergic agonist, present in both extracellular and intracellular fluids, with promising cytoprotective effects in different diseases including neurodegenerative processes. In a search for efficient neuroprotective strategies for SCI, we have tested the capability of Ap₄A to reduce the excitotoxic death mediated by the ATP-induced deregulation of calcium homeostasis and its consequences on tissue preservation and functional recovery in a mouse model of moderate contusive SCI. Our analyses with the murine neural cell line Neuro2a demonstrate that treatment with Ap₄A reduces ATP-dependent excitotoxic death by both lowering the intracellular calcium response and decreasing the expression of specific purinergic receptors. Follow-up analyses in a mouse model of contusive SCI showed that acute administration of Ap₄A following SCI reduces tissue damage and improves motor function recovery. These results suggest that Ap₄A cytoprotection results from a decrease of the purinergic tone preventing the effects of a massive release of ATP after SCI, probably together with a direct induction of anti-apoptotic and pro-survival pathways via activation of P2Y₂ proposed in previous studies. In conclusion, Ap₄A may be a good candidate for an SCI therapy, particularly to reduce excitotoxicity in combination with other modulators and/or inhibitors of the excitotoxic process that are being tested.