Feedback-Controlled Release of Alendronate from Composite Microparticles.

Extended bone fractures or fractures coexisting with bone disorders can lead to non-unions where surgical intervention is required. Composite drug delivery systems are being used increasingly more in order to treat such defects locally. Alendronate (ALD), a bisphosphonate extensively used in clinical practice to treat conditions, such as osteoporosis, has been shown to assist bone fracture healing through its antiresorptive capacity. This study reports the development of a polymeric composite system for the in situ delivery of ALD, which possesses enhanced encapsulation efficiency (EE%) and demonstrates controlled release over a 70-day period. ALD and calcium phosphate (CaP) were incorporated within poly (lactic-co-glycolic acid) (PLGA) microspheres, giving rise to a 70% increase in EE% compared to a control system. Finally, a preliminary toxicological evaluation demonstrated a positive effect of the system on pre-osteoblastic cells over 72 h.

Orchid Bee (Apidae: Euglossini) Communities in Atlantic Forest Remnants and Restored Areas in Paraná State, Brazil.

In this study, we compare orchid bee communities surveyed in four forest remnants of the Atlantic Forest and four reforested areas characterized by seasonal semi-deciduous forest vegetation in different successional stages (mature and secondary vegetation), located in southern Brazil. The sizes of forest remnants and reforested areas varied from 32.1 to 583.9 ha and from 11.3 to 33.3 ha, respectively. All reforested areas were located near one forest remnant. During samplings, totaling nine per study area, euglossine males were attracted to eight scent baits and captured with bait trap and entomological nets. Each forest remnant and its respective reforested area were sampled simultaneously by two collectors. We collected 435 males belonging to nine species of orchid bees distributed in four genera. The number of individuals and species did not differ significantly between different areas, except for a reforested area (size 33.3 ha), which was located far from its respective forest remnant. Our findings also revealed an apparent association between an orchid bee species (Euglossa annectans Dressler 1982) and the most preserved area surveyed in our study, suggesting that this bee is a potential indicator of good habitat quality in recuperating or preserved areas. Our results suggest that reforested habitats located near forest remnants have a higher probability of having reinstated their euglossine communities.

Fine-scale genetic structure patterns in two freshwater fish species, Geophagus brasiliensis (Osteichthyes, Cichlidae) and Astyanax altiparanae (Osteichthyes, Characidae) throughout a Neotropical stream.

Streams are very important environments for Neotropical freshwater fish fauna, and possess a high number of species. These small drainages are also highlighted by their intrinsic biological and physicochemical features; however, knowledge on the genetic distribution of fish in these drainages is limited. Therefore, in the present study, RAPD (random amplified polymorphic DNA) and microsatellite markers were used to analyze population differentiation and gene flow of Astyanax altiparanae and Geophagus brasiliensis from three sites (high, medium, and low) throughout the Penacho stream (about 32 km long), which is a Neotropical stream. Both markers revealed higher levels of genetic diversity levels for A. altiparanae (: 90.05; HS: 0.350) compared to G. brasiliensis (: 30.43; HS: 0.118), which may be related to the particular biology of each species. AMOVA revealed significant genetic variation among populations of each species. All pairwise ΦST values were significant, ranging from 0.020 to 0.056 for A. altiparanae samples, and from 0.065 to 0.190 for G. brasiliensis samples. Bayesian clustering analysis corroborated these results and revealed clusters of both A. altiparanae (two based on RAPD data) and G. brasiliensis (two based on RAPD data and three on microsatellite data). Gene flow estimates showed that there were similar rates of migration among A. altiparanae samples and low rates of migration among some G. brasiliensis samples. These results suggest patterns of fine-scale genetic structure for both species in the Penacho stream. This information may enhance knowledge of Neotropical streams and may be useful for future management and conservation activities.

Isolation and characterization of 13 polymorphic microsatellite loci in Hypostomus ancistroides (Teleostei, Loricariidae) and cross-amplification in related species.

This study details 13 novel polymorphic microsatellite loci in the armoured catfish Hypostomus ancistroides, and assesses their utility for population genetic studies. The analysis of 30 individuals revealed a total of 99 different alleles (ranging from two to 15 alleles per locus), with an average of 7·62 alleles per locus, with observed and expected heterozygosities ranging from 0·103 to 0·931 and from 0·102 to 0·906, respectively. One of the 13 loci showed significant deviations from Hardy-Weinberg equilibrium, probably due to the presence of null alleles, inferred from the excess of homozygotes.

DNA barcoding of freshwater ichthyoplankton in the Neotropics as a tool for ecological monitoring.

Quantifying and classifying ichthyoplankton is one of the most effective ways of monitoring the recruitment process in fishes. However, correctly identifying the fish based on morphological characters is extremely difficult, especially in the early stages of development. We examined ichthyoplankton from tributaries and reservoirs along the middle stretch of the Paranapanema River, one of the areas most impacted by hydroelectric projects in the Neotropics. Matching DNA sequences of the COI gene (628-648 bp) allowed us to identify 99.25% of 536 samples of eggs (293) and larvae (243) subjected to BOLD-IDS similarity analysis with a species-level threshold of 1.3%. The results revealed 37 species in 27 genera, 15 families and four orders, some 23.8% of documented fish species in the Paranapanema River. Molecular identification meant that we could include data from egg samples that accounted for about 30% of the species richness observed. The results in this study confirm the efficacy of DNA barcoding in identifying Neotropical ichthyoplankton and show how the data produced provide valuable information for preparing plans for conserving and managing inland waters.

A Comparison of Social Bee-Plant Networks between Two Urban Areas.

In the last decade, several studies demonstrated the effectiveness of ecological network analysis to a better understanding of the structure bee-plant interaction networks; however, such approaches involving urban areas are still scarce. Here, we analyzed two assemblages of corbiculate bees (Apoidea, Apidae) in two geographically distinct urban areas in Brazil. In both study areas, apid bees visiting flowers were captured with an insect net. Surveys were performed biweekly and alternately in each area, over a 1-year period. Both urban areas were very similar for most indices. The two social bee-plant networks were significantly nested, a pattern usually described for bee-plant networks and somehow expected in our study, considering the recognized behavior of social apid bees in exploring a wide range of plant species. The modularity measures were low and very similar for the networks of both urban areas, a finding that could be due at least in part to the low phylogenetic distance between corbiculate bees and the broad dietary habits of the social apid bees. Network-level indices showed that both bee assemblages had a relatively low niche overlap, indicating that the set of social apid species studied exploited differently the arrays of plants available. Species level index (resource range) showed that in both urban areas, Trigona spinipes (Fabr.) and Apis mellifera L. showed the higher number of interactions, a result that demonstrates the importance of these species in social bee-plant interaction networks in urban areas. Similarly to other ecosystems, these two apid species behaved as super-generalists in the two urban areas surveyed herein.

Development and characterization of 14 microsatellite loci in the Neotropical fish Geophagus brasiliensis (Perciformes, Cichlidae).

Fourteen polymorphic microsatellite loci were isolated and characterized for the Neotropical cichlid Geophagus brasiliensis and tested on 30 individuals belonging to a single population. Among the 14 loci described, four showed potential presence of null alleles, inferred from the excess of homozygous genotypes, and three of these loci showed significant deviations from Hardy-Weinberg equilibrium. Fifty-nine different alleles were detected (ranging from two to eight alleles per locus), with estimates of observed and expected heterozygosity ranging from 0·167 to 0·700 and from 0·269 to 0·825. Cross-amplification of primers was successful in five other cichlid species.

In vivo and in vitro exposures for the evaluation of the genotoxic effects of lead on the Neotropical freshwater fish Prochilodus lineatus.

In the present study, in vivo and in vitro exposures were used to assess the genotoxicity of lead (Pb) to the freshwater fish Prochilodus lineatus. The comet assay using blood, liver and gill cells, and the occurrence of micronuclei (MN) and other erythrocytic nuclear abnormalities (ENA) were used to assess the genotoxic potential of lead in vivo. Metallothionein content (MT) was measured in fish liver in order to evaluate the protection of fish against Pb toxicity. Fish erythrocytes were exposed to Pb in vitro (1, 3 and 6 h) and the number of viable cells, DNA integrity, using the comet assay, and lysosomal membrane stability, measured by the neutral red retention assay (NRRA) were analyzed. The results of the comet assay after in vivo toxicity tests (6, 24 and 96 h) showed that Pb was genotoxic for all the three tissues analyzed after 96 h exposure. A significant increase in liver MT content was observed after 6 and 24 h of Pb exposure. MN frequency did not increase after Pb exposures, but the frequency of the other ENA, such as kidney-shaped nuclei, segmented nuclei and lobed nuclei, showed a significant increase after 24 and 96 h, indicating that ENA is a better biomarker for Pb exposure than MN alone after short-term exposures. The results of the comet assay performed with erythrocytes in vitro exposed to lead confirmed its genotoxic effect and showed that DNA damage increased with increasing exposure time. Moreover, the NRRA clearly indicated that Pb induces a destabilization of the lysosomal membrane. These results demonstrate the potential genotoxicity and cytotoxicity of lead after acute exposures.

Genetic damage in the bivalve mollusk Corbicula fluminea induced by the water-soluble fraction of gasoline.

Although gasoline is an important contaminant of aquatic ecosystems, information concerning the potential effects of this petroleum derivative on the DNA of aquatic biota is lacking. The present study aimed to evaluate the genotoxic and mutagenic effects of acute exposures (6, 24, and 96h) to gasoline water-soluble fraction (GWSF), diluted to 5%, on the Asian clam Corbicula fluminea. The comet assay and the micronucleus (MN) test were performed on hemocytes and gill cells of C. fluminea. For the three different times tested, the comet assay indicated DNA damage in hemocytes and gill cells of C. fluminea exposed to GWSF. The MN test detected significant damage in the genetic material of the hemocytes only after 96h of exposure to GWSF. The recovery capacity of organisms previously exposed for 6h to GWSF was also evaluated with the comet assay. The results revealed a great capacity of this species to repair DNA damage; following 6h of recovery, the comet score returned to that of the control groups. Overall, our findings indicate that GWSF has genotoxic and mutagenic effects on C. fluminea. In addition, the present results confirm the sensitivity of C. fluminea to damage caused by exposure to environmental contaminants, and, therefore, its suitability for use in environmental monitoring studies.

Genotoxic effects of Roundup on the fish Prochilodus lineatus.

Glyphosate-based herbicides, such as Roundup, represent the most extensively used herbicides worldwide, including Brazil. Despite its extensive use, the genotoxic effects of this herbicide are not completely understood and studies with Roundup show conflicting results with regard to the effects of this product on the genetic material. Thus, the aim of this study was to evaluate the genotoxic effects of acute exposures (6, 24 and 96 h) to 10 mg L(-1) of Roundup on the neotropical fish Prochilodus lineatus. Accordingly, fish erythrocytes were used in the comet assay, micronucleus test and for the analysis of the occurrence of nuclear abnormalities and the comet assay was adjusted for branchial cells. The results showed that Roundup produces genotoxic damage in erythrocytes and gill cells of P. lineatus. The comet scores obtained for P. lineatus erythrocytes after 6 and 96 h of exposure to Roundup were significantly higher than respective negative controls. For branchial cells comet scores were significantly higher than negative controls after 6 and 24 h exposures. The frequencies of micronucleus and other erythrocyte nuclear abnormalities (ENAs) were not significantly different between Roundup exposed fish and their respective negative controls, for all exposure periods. In conclusion, the results of this work showed that Roundup produced genotoxic effects on the fish species P. lineatus. The comet assay with gill cells showed to be an important complementary tool for detecting genotoxicity, given that it revealed DNA damage in periods of exposure that erythrocytes did not. ENAs frequency was not a good indicator of genotoxicity, but further studies are needed to better understand the origin of these abnormalities.