Communities of endophytic microorganisms in different developmental stages from a local variety as well as transgenic and conventional isogenic hybrids of maize

The diversity of endophytic microorganisms may change due to the genotype of the host plant and its phenological stage. In this study we evaluated the effect of phenological stage, transgenes and genetic composition of maize on endophytic bacterial and fungal communities. The maize populations were composed of a local variety named Rosado (RS) and three isogenic hybrids. One isogenic hybrid was not genetically modified (NGM). Another hybrid (Hx) contained the transgenes cry1F and pat (T1507 event), which provide resistance to insects of the order Lepidoptera and tolerance to the glufosinate-ammonium herbicide, respectively. The third hybrid (Hxrr) contained the transgene cp4 epsps (NK603 event) combined with the transgenes cry1F and pat (T1507 event), which allow tolerance to the Roundup Ready herbicide, besides the characteristics of Hx. Evaluation of the foliar tissue was done through PCR-DGGE analysis, with specific primers for bacteria and fungi within four phenological stages of maize. The endophytic bacteria were only clustered by phenological stages; the structure of the fungal community was clustered by maize genotypes in each phenological stage. The fungal community from the local variety RS was different from the three hybrids (NGM, Hx and Hxrr) within the four evaluated stages. In the reproductive stage, the fungal community from the two transgenic hybrids (Hx and Hxrr) were separated, and the Hxrr was different from NGM, in the two field experiments. This research study showed that the genetic composition of the maize populations, especially the presence of transgenes, is the determining factor for the changes detected in the endophytic fungal community of maize leaves.     

Synthesis,antiplatelet and antithrombotic activities of resveratrol derivatives with NO-donor properties

Resveratrol (RVT) is a stilbene with a protective effect on the cardiovascular system; however, drawbacks including low bioavailability and fast metabolism limit its efficacy. In this work we described new resveratrol derivatives with nitric oxide (NO) release properties, ability to inhibit platelet aggregation and in vivo antithrombotic effect. Compounds (4a–f) were able to release NO in vitro, at levels ranging from 24.1% to 27.4%. All compounds (2a–f and 4a–f) have exhibited platelet aggregation inhibition using as agonists ADP, collagen and arachidonic acid. The most active compound (4f) showed reduced bleeding time compared to acetylsalicylic acid (ASA) and protected up to 80% against in vivo thromboembolic events. These findings suggest that hybrid resveratrol-furoxan (4f) is a novel lead compound able to prevent platelet aggregation and thromboembolic events.     

Length–weight relationships of 18 fish species from Paraíba do Sul basin,Minas Gerais,Brazil

The present study established the length–weight relationships (LWRs) for 18 species of freshwater fish from Paraíba do Sul River basin, southeastern Brazil. LWR parameters are estimated for the first time for seven of these species. The allometric coefficient b varied from 2.81 to 3.32. These data complement the effort to understand the biology of one of the most threatened Brazilian basins.     

Carbofuran-induced alterations in biochemical composition, lipoperoxidation, and Na+/K+ATPase activity of Hyalella pleoacuta and Hyalella curvispina in bioassays

The present study investigated the effects of carbofuran on the energy metabolism (levels of glycogen, total proteins, total lipids, triglycerides, and lipoperoxidation), Na+/K+ATPase activity, and reproductive parameters (formation of couples, ovigerous females, and mean number of eggs) in the freshwater amphipods Hyalella pleoacuta and Hyalella curvispina. These crustaceans live in limnetic environments of the plateau (H. pleoacuta) and coastal plain (H. curvispina) of the state of Rio Grande do Sul in southern Brazil. The animals were collected in the winter of 2006 in the Vale das Trutas (28 degrees 47'00'S-49 degrees 50'53'W) in the Municipality of S?o José dos Ausentes, and in Gentil Lagoon (29 degrees 56'30'S, 50 degrees 07'50'W) in the Municipality of Tramandaí. In the laboratory, the amphipods were kept submerged in aquariums under controlled conditions of photoperiod (12 h light: 12 h dark), temperature (23 degrees C+/-1), and constant oxygenation. Animals were exposed to carbofuran at a dose of 5 or 50 microg/L for a period of 7 days. At the end of this period, the animals were immediately frozen for determination of the biochemical parameters, lipoperoxidation levels (TBARS), and enzyme Na+/K+ATPase activity. During each day of culture, several reproductive parameters were observed. Statistical analysis (ANOVA) revealed that carbofuran induces significant decreases in glycogen, proteins, lipids, triglycerides, and Na+/K+ATPase, as well as a significant increase in lipoperoxidation levels. Studies of all the biochemical parameters seem to be quite promising, in order to assess and predict the effects of toxicants on non-target organisms. The results also suggest that the reproductive parameters (formation of couples, ovigerous females and mean number of eggs) may provide sensitive criteria for assessing ecotoxicological effects. Furthermore, H. pleoacuta and H. curvispina are suitable organisms for use in toxicity tests, and we suggest that they are sensitive species that could be used in monitoring studies.     

Asymbiotic seed germination and in vitro seedling development of <Emphasis Type="Italic">Cyrtopodium punctatum</Emphasis>: a propagation protocol for an endangered Florida native orchid

Cyrtopodium punctatum Lindley is an endangered epiphytic orchid restricted in the United States to southern Florida. Due to its ornamental value, the species was extensively collected from the wild during the past 100 years. Today, only a few plants remain in protected areas. As part of a conservation plan for the species, procedures for asymbiotic seed germination were developed. Five asymbiotic orchid seed germination media (PhytoTechnology Orchid Seed Sowing Medium, Knudson C, Malmgren Modified Terrestrial Orchid Medium, Vacin &; Went Modified Orchid Medium, and ½-strengh Murashige &; Skoog) were examined for their effectiveness in promoting seed germination and protocorm development under a 16/8 h L/D photoperiod and dark (0/24 h L/D). The influence of photoperiod on growth and development was also examined. Seeds were germinated under a 16/8 h, 12/12 h, 8/16 h L/D photoperiod, at 25 ± 3°C and allowed to develop in vitro for 10 weeks. After 10 weeks, developing seedlings were transferred to Sigma Phytatrays and returned to their assigned photoperiod treatments for continued seedling development for an additional 15 weeks. Highest germination occurred in 0/24 h L/D on PhytoTechnology Orchid Seed Sowing Medium and seedlings displayed more advanced development when cultured under 16/8 h L/D photoperiod after 15 weeks in Phytatrays. Thirty-five week old seedlings potted in coconut husk growing medium exhibited 90% survival following 5 weeks acclimatization to greenhouse conditions. This asymbiotic seed germination protocol for C. punctatum will facilitate future reintroduction projects involving this endangered species.     

Effect of organic tomato (Lycopersicon esculentum) extract on the genotoxicity of doxorubicin in the Drosophila wing spot test

The consumption of organic tomatoes (ORTs) reduces the risk of harmful effects to humans and the environment caused by exposure to toxic agrochemicals. In this study, we used the somatic mutation and recombination test (SMART) of wing spots in Drosophila melanogaster to evaluate the genotoxicity of ORT and the effect of cotreatment with ORT on the genotoxicity of Doxorubicin^® (DXR, a cancer chemotherapeutic agent) that is mediated by free radical formation. Standard (ST) cross larvae were treated chronically with solutions containing 25%, 50% or 100% of an aqueous extract of ORT, in the absence and presence of DXR (0.125 mg/mL), and the number of mutant spots on the wings of emergent flies was counted. ORT alone was not genotoxic but enhanced the toxicity of DXR when administered concomitantly with DXR. The ORT-enhanced frequency of spots induced by DXR may have resulted from the interaction of ORT with the enzymatic systems that catalyze the metabolic detoxification of this drug.     

Carbofuran-induced alterations in the energy metabolism and reproductive behaviors of Hyalella castroi (Crustacea,Amphipoda)

Continuous or pulse exposure to pesticides may have negative effects on non-target organisms, resulting in a number of pathological and disturbed biochemical processes, including changes in energy budgets. The objective of this investigation was to examine the potential effects of carbofuran on the biochemical composition (glycogen, proteins, total lipids, triglycerides and cholesterol), levels of lipoperoxidation, Na⁺/K⁺ATPase activity, and reproductive behaviors (number of reproductive pairs, ovigerous females, and number of eggs) in the amphipod Hyalella castroi. The amphipods were collected in spring 2007, in the southern Brazilian highlands. In the laboratory, the animals were kept in aquariums under controlled conditions for 7 days, and after this period were exposed to 1 or 10 µg/L of carbofuran for 7 days. After the period of exposure, the animals were immediately frozen for determination of glycogen, proteins, lipids, triglycerides, total cholesterol, levels of lipoperoxidation, and Na⁺/K⁺ATPase activity. During each day of culture, reproductive behaviors were observed. Carbofuran induced significant decreases in biochemical reserves (glycogen, proteins, lipids, triglycerides and cholesterol), a significant increase in lipoperoxidation levels, and a decrease in Na⁺/K⁺ATPase activity in both males and females. Studies of all the biochemical parameters seem to be quite promising, in order to assess and predict the effects of toxicants on non-target organisms. The results showed that reproductive behaviors may provide sensitive criteria for assessing ecotoxicological effects. H. castroi lives among rooted aquatic macrophytes, and we suggest that it is a sensitive species that could be used in monitoring studies.     

5-Heteroatom-substituted pyrazoles as canine COX-2 inhibitors: Part 2. Structure-activity relationship studies of 5-alkylethers and 5-thioethers

Structure-activity relationship (SAR) studies of novel 2-[3-trifluoromethyl-5-alkyl(thio)ether pyrazo-1-yl]-5-methanesulfonyl pyridine derivatives for canine COX enzymes are described. The 4-cyano-5-alkyl ethers were found to have excellent potency and selectivity, whereas the 5-thioethers were potent but less selective than the ether analogs in a canine whole blood (CWB) COX-2 assay.     

Biocatalytic characterization of Aspergillus oryzae β-galactosidase immobilized on functionalized multi-walled carbon nanotubes

The objectives of this work were to immobilize commercial Aspergillus oryzae β-galactosidase on functionalized multi-walled carbon nanotubes (MWCNTs) using different treatments and to characterize the products. Treatments were performed with glutaraldehyde, ethylenediamine and a mixture of concentrated H₂SO₄:HNO₃. The MWCNTs and their derivatives were characterized by thermogravimetric analysis. The immobilized enzymes were evaluated using inactivation kinetics, operating conditions, that is pH and temperature, kinetic parameters and lactose hydrolysis reusability. Immobilization yield and efficiency were significantly higher for β-galactosidase immobilized on MWCNTs functionalized by the acid mixture (Ac-Gal-MWCNTs). These values were 97% and 82%, respectively, after 3?h of immobilization. The activity of the Ac-Gal-MWCNTs was maintained at ～51% of their initial activity after being stored for 90 days at 4?°C. The Ac-Gal-MWCNTs retained more than 90% of their initial activity up to the fourth recycle. As the acid functionalization was the most efficient method tested for immobilizing A. oryzae β-galactosidase on MWCNTs, this method shows promise for industrial applications.     

Biochar Enhances Aspergillus niger Rock Phosphate Solubilization by Increasing Organic Acid Production and Alleviating Fluoride Toxicity

During fungal rock phosphate (RP) solubilization, a significant quantity of fluoride (F⁻) is released together with phosphorus (P), strongly inhibiting the process. In the present study, the effect of two F⁻ adsorbents [activated alumina (Al₂O₃) and biochar] on RP solubilization by Aspergillus niger was examined. Al₂O₃ adsorbed part of the F⁻ released but also adsorbed soluble P, which makes it inappropriate for microbial RP solubilization systems. In contrast, biochar adsorbed only F⁻ while enhancing phosphate solubilization 3-fold, leading to the accumulation of up to 160 mg of P per liter. By comparing the values of F⁻ measured in solution at the end of incubation and those from a predictive model, it was estimated that up to 19 mg of F⁻ per liter can be removed from solution by biochar when added at 3 g liter⁻¹ to the culture medium. Thus, biochar acted as an F⁻ sink during RP solubilization and led to an F⁻ concentration in solution that was less inhibitory to the process. In the presence of biochar, A. niger produced larger amounts of citric, gluconic, and oxalic acids, whether RP was present or not. Our results show that biochar enhances RP solubilization through two interrelated processes: partial removal of the released F⁻ and increased organic acid production. Given the importance of organic acids for P solubilization and that most of the RPs contain high concentrations of F⁻, the proposed solubilization system offers an important technological improvement for the microbial production of soluble P fertilizers from RP.