Structural basis for the interaction of isoDGR with the RGD-binding site of alphavbeta3 integrin

Asparagine deamidation at the NGR sequence in the 5th type I repeat of fibronectin (FN-I5) generates isoDGR, an alphavbeta3 integrin-binding motif regulating endothelial cell adhesion and proliferation. By NMR and molecular dynamics studies, we analyzed the structure of CisoDGRC (isoDGR-2C), a cyclic beta-peptide mimicking the FN-I5 site, and compared it with NGR, RGD, or DGR-containing cyclopeptides. Docking experiments show that isoDGR, exploiting an inverted orientation as compared with RGD, favorably interacts with the RGD-binding site of alphavbeta3, both recapitulating canonical RGD-alphavbeta3 contacts and establishing additional polar interactions. Conversely, NGR and DGR motifs lack the fundamental pharmacophoric requirements for high receptor affinity. Therefore, unlike NGR and DGR, isoDGR is a new natural recognition motif of the RGD-binding pocket of alphavbeta3. These findings contribute to explain the different functional properties of FN-I5 before and after deamidation, and provide support for the hypothesis that NGR --> isoDGR transition can work as a molecular timer for activating latent integrin-binding sites in proteins, thus regulating protein function.     

Protection against hepatitis B by the Butang recombinant vaccine in newborn children in South Brazil

The prevention of hepatitis B by vaccination is one of the most efficient tools to avoid the transmission of the virus. This study evaluated the immunogenicity of the national vaccine Butang in children born in Campo Mour?o City, state of Paraná, Brazil, aged 7 to 12 months, by determining the anti-HBsAg antibodies levels after completion of the National Immunization Program Protocol for hepatitis B. All 70 children evaluated by the MEIA method (immune-enzymatic micro particles) showed seroconversion to the Butang vaccine. Nine children (12.9%) presented a low response, with anti-HBs titers between 11 and 100 mUI/ml; 39 children (55.7%) showed a good response to the vaccine, with titers between 101 and 1000 mUI/ml; and 22 children (31.4%) showed antibodies titers higher than 1000 mUI/ml. The mean titer of the anti-HBs antibody titers was 1408.1 +/- 2870.26 mUI/ml (15.7 to 19560.0 m UI/ml). The levels of antibodies produced by the prematurely-born children were not statistically different from those found in the newborns. Fifty-five children were also evaluated through the ELFA method (ELISA with a final detection in fluorescence), which presented similar results. The results obtained in our study corroborated the effectiveness of the national vaccine Butang in newborn children of Campo Mour?o City, Paraná, even if they were premature.     

Vascular endothelial growth factor stimulates organ-specific host matrix metalloproteinase-9 expression and ovarian cancer invasion

Vascular endothelial growth factor (VEGF) and matrix metalloproteinases (MMP) regulate each other, contributing to tumor progression. We have previously reported that MMP9 induces the release of tumor VEGF, promoting ascites formation in human ovarian carcinoma xenografts. The aim of this study was to investigate whether tumor-derived VEGF regulated the expression of gelatinase by the stroma, influencing the invasive properties of ovarian tumors. Tumor variants derived from 1A9 human ovarian carcinoma, stably expressing VEGF(121) in the sense (1A9-VS-1) and antisense orientations (1A9-VAS-3), were used. In vivo, zymographic analysis of tumors from 1A9-VS-1 implanted in the peritoneal cavity of nude mice showed higher levels of gelatinases, particularly murine MMP9, indicating that VEGF stimulates host expression of the matrix-degrading enzyme. Murine MMP9 expression was also high in the ovaries of mice bearing 1A9-VS-1 tumors. The effect on host MMP9 activity was organ-specific. The levels of host pro-MMP9 in ovaries correlated with the plasma levels of tumor VEGF and with the selective invasion of the ovaries. Induction of host MMP9 expression in tumors and ovaries was independent of the site of tumor growth as it was seen in mice carrying both intraperitoneal and subcutaneous tumors. The anti-VEGF antibody bevacizumab (Avastin) inhibited MMP9 expression and tumor invasion in the ovaries of mice bearing 1A9-VS-1 tumors. These findings point to a complex cross-talk between VEGF and MMPs in the progression of ovarian tumor and suggest the possibility of using VEGF inhibitors to affect MMP-dependent tumor invasion.     

Selective and efficient immunoprecipitation of the disease-associated form of the prion protein can be mediated by nonspecific interactions between monoclonal antibodies and scrapie-associated fibrils

Transmissible spongiform encephalopathies are characterized by the accumulation in brain tissues of an abnormal isoform of the prion protein named PrPsc, which is the only direct marker known for transmissible spongiform encephalopathies. Here we show that PrPsc can be specifically immunoprecipitated by using several monoclonal antibodies (mAbs) of various specificities independently of the properties of their binding site (paratope). These results strongly suggest that a significant proportion of mAbs can interact with PrPsc aggregates through nonspecific paratope-independent interactions allowing selective immunoprecipitation of PrPsc when these mAbs are immobilized on a polydisperse solid phase like microbeads.     

Isolation of an effective strain of Frankia from nodules of Discaria trinervis (Rhamnaceae)

Frankia strain BCU110501 was isolated from root nodules of the native Patagonian actinorhizal plant Discaria trinervis. The strain was grown on BAP medium with sodium propionate or glucose as carbon sources. Colonies grown in nitrogen-free medium showed branched hyphae bearing polymorphic sporangia and vesicles, which were capable of nitrogen fixation. Old cultures produced a red pigment. The infectivity and effectivity of a Frankia strain isolated from Discaria on its own host, D. trinervis and also in D. chacaye, is reported for the first time. Frankia BCU110501 has physiological properties that are intermediate between categories proposed by Lechevalier et al. (1983) to classify Frankia.     

Influence of propionate on growth and fermentative activity of lactobacilli

Propionic acid was added to cultures of Lactobacillus helveticus and Lact. casei growing in a complex medium. The effect of different concentrations of this fatty acid on the growth kinetic, substrate consumption rate and coefficient and product formation rate was examined. Low concentrations of propionic acid produced a low increase or no modification of the growth rate of lactobacilli. By contrast, higher concentrations reduced the growth rate and substrate yields in all the strains. The fermentative activity of lactobacilli increased in all the cultures with propionate. When the concentration of glucose in the medium was low, the population density of all the strains at the stationary growth phase was lower in the presence of propionate than in control cultures. An increase in the concentration of glucose in the medium counteracted the inhibitory effect of propionate. The addition of lithium lactate to the medium gave greater inhibition even at low propionate concentrations. The inhibition results were explained by assuming that propionic acid could diffuse through cell membranes in the undissociated form, increasing the inward leak of H⁺ in the cells. The extrusion of H⁺ by the H⁺-ATPase is possible by increasing the fermentative activity of the cells. When metabolism cannot supply the ATP required for proton extrusion, growth rate and biomass production decrease.     

Water and fish select for fleshy fruits in tropical wetland forests

Adjacent floodplain and upland tropical forests experience the same temperature and precipitation regimes, but differ substantially in plant species composition and biotic interactions because of extensive flooding. We hypothesize that flooded forests filter fruiting traits linked to seed dispersal by water and fishes, such that selection by water and fish led to (1) trees that synchronize the timing of fruiting with annual floods, and (2) the evolution of fleshy tissues on fruits to improve buoyancy and increase fruit consumption rates by fish. To test this hypothesis, we compared plant communities in seasonally flooded forests and adjacent upland forest in terms of fruiting phenology, the frequency of trees bearing fleshy fruit, and the role of fleshy tissues in buoyancy and seed viability. Beta‐diversity in this system is high, with significant differences in species composition across habitats. As predicted, the production of ripe fleshy fruits was significantly greater in flooded than upland forests during the flood season. Furthermore, we found that trees with fleshy fruit were significantly more abundant in flooded forests even though species richness of fleshy fruit‐bearing trees was proportionally similar in flooded and upland forests. Additionally, fleshy pulp increased buoyancy. Likewise, time afloat decreased for denser fruit and those with high seed to pulp ratios. In concert, these results suggest that fleshy fruits in Neotropical floodplain forests facilitated hydrochory and ichthyochory. Once established, water and fish became important agents of selection on fruiting traits.