Temporal and tissue expression of genes involved in buds of earliness cotton cultivar.

Gene sequences previously identified in Arabidopsis buds were used as references in order to estimate temporal and tissue expression in buds, leaves, stem, and root tissues in cotton plants. Buds were evaluated during 3 phases: 2-8, 10-12, and 14-20 mm. Primers were designed for the ARF6, ATFY, and SEUSS genes for use in semi-quantitative reverse transcription-polymerase chain reaction and quantitative reverse transcription-polymerase chain reaction. Different levels of expression of the 3 genes were confirmed in cotton buds as well as in other tissues. The peak of gene expression was observed in buds sized 10-12 mm, after which expression decreased in larger buds. The gene GhFYPP3 was the most promising for further prospection of promoter regions, with regular expression patterns observed in bud sizes 10-12 and 14-20 mm. This trait was not observed in others genes.

Genes expressed in cotton (Gossypium hirsutum) buds isolated with a subtractive library.

A subtractive cDNA library from cotton buds was constructed to prospect for differentially expressed genes related to early bud development. A library was constructed and 768 cDNA sequences were obtained, comprising 168 clusters, with 126 contigs and 42 singlets. Both the Gossypium as well as Arabidopsis databases were utilized for the in silico analysis, since some genes identified in cotton have not yet been studied for functionality, although they have homology with genes from other species. The transcriptome revealed a large number of transcripts, some of them with unknown function, and others related to pollen development, pollen tubes, ovules, and fibers at different stages. The most populated contig was identified as fiber from 0-10 days after anthesis, with 12 reads. The success and novelty rates generated from the library were 67 and 51%, respectively. The information obtained here will provide a framework for research on functional cotton genomics.

Dero (Allodero) lutzi Michaelsen, 1926 (Oligochaeta: Naididae) associated with Scinax fuscovarius (Lutz, 1925) (Anura: Hylidae) from Semi-deciduous Atlantic Rain Forest, southern Brazil.

Amphibians are hosts for a wide variety of ecto- and endoparasites, such as protozoans and parasitic worms. Naididae is a family of Oligochaeta whose species live on a wide range of substrates, including mollusks, aquatic macrophytes, sponges, mosses, liverworts, and filamentous algae. However, some species are known as endoparasitic from vertebrates, such as Dero (Allodero) lutzi, which is parasitic of the urinary tracts of frogs, but also have a free-living stage. Specimens in the parasitic stage lack dorsal setae, branchial fossa, and gills. Here we report the occurrence of D. (A.) lutzi associated with anuran Scinax fuscovarius from Semi-deciduous Atlantic Rain Forest in southern Brazil. The study took place at the Caiuá Ecological Station, Diamante do Norte, Paraná, southern Brazil. Seven specimens of S. fuscovarius were examined for parasites but only one was infected. Parasites occurred in ureters and urinary bladder. Previous records of this D. (A.) lutzi include the Brazilian States of Santa Catarina, São Paulo, Rio de Janeiro, and Minas Gerais, as well as Cuba and North America. This is a new locality record for this species in Brazil. Reports of Dero (Allodero) lutzi are rare, due to difficulty of observation, and such events are restricted only the fortuitous cases. It is important to emphasize the necessity of future studies, which are fundamental to the understanding of biological and ecological aspects of this species.

Modulation of peritoneal macrophage activity by the saturation state of the fatty acid moiety of phosphatidylcholine.

To determine the effects of saturated and unsaturated fatty acids in phosphatidylcholine (PC) on macrophage activity, peritoneal lavage cells were cultured in the presence of phosphatidylcholine rich in saturated or unsaturated fatty acids (sat PC and unsat PC, respectively), both used at concentrations of 32 and 64 microM. The treatment of peritoneal macrophages with 64 microM unsat PC increased the production of hydrogen peroxide by 48.3% compared to control (148.3 +/- 16.3 vs 100.0 +/- 1.8%, N = 15), and both doses of unsat PC increased adhesion capacity by nearly 50%. Moreover, 64 microM unsat PC decreased neutral red uptake by lysosomes by 32.5% compared to the untreated group (67.5 +/- 6.8 vs 100.0 +/- 5.5%, N = 15), while both 32 and 64 microM unsat PC decreased the production of lipopolysaccharide-elicited nitric oxide by 30.4% (13.5 +/- 2.6 vs 19.4 +/- 2.5 microM) and 46.4% (10.4 +/- 3.1 vs 19.4 +/- 2.5 microM), respectively. Unsat PC did not affect anion production in non-stimulated cells or phagocytosis of unopsonized zymosan particles. A different result pattern was obtained for macrophages treated with sat PC. Phorbol 12-miristate 13-acetate-elicited superoxide production and neutral red uptake were decreased by nearly 25% by 32 and 64 microM sat PC, respectively. Sat PC did not affect nitric oxide or hydrogen peroxide production, adhesion capacity or zymosan phagocytosis. Thus, PC modifies macrophage activity, but this effect depends on cell activation state, fatty acid saturation and esterification to PC molecule and PC concentration. Taken together, these results indicate that the fatty acid moiety of PC modulates macrophage activity and, consequently, is likely to affect immune system regulation in vivo.

Modulation of peritoneal macrophage activity by the saturation state of the fatty acid moiety of phosphatidylcholine

To determine the effects of saturated and unsaturated fatty acids in phosphatidylcholine (PC) on macrophage activity, peritoneal lavage cells were cultured in the presence of phosphatidylcholine rich in saturated or unsaturated fatty acids (sat PC and unsatPC, respectively), both used at concentrations of 32 and 64 ìM. The treatment of peritoneal macrophages with 64 ìM unsat PC increased the production of hydrogen peroxide by 48.3% compared to control (148.3 ± 16.3 vs 100.0 ± 1.8%, N = 15), and both doses of unsat PC increased adhesion capacity by nearly 50%. Moreover, 64 ìM unsat PC decreased neutral red uptake by lysosomes by 32.5% compared to the untreated group (67.5 ± 6.8 vs 100.0 ± 5.5%, N = 15), while both 32 and 64 ìM unsat PC decreased the production of lipopolysaccharide-elicited nitric oxide by 30.4% (13.5 ± 2.6 vs 19.4 ± 2.5 ìM) and 46.4% (10.4 ± 3.1 vs 19.4 ± 2.5 ìM), respectively. Unsat PC did not affect anion production in non-stimulated cells or phagocytosis of unopsonized zymosan particles. A different result pattern was obtained for macrophages treated with sat PC. Phorbol 12-miristate 13-acetate-elicited superoxide production and neutral red uptake were decreased by nearly 25% by 32 and 64 ìM sat PC, respectively. Sat PC did not affect nitric oxide or hydrogen peroxide production, adhesion capacity or zymosan phagocytosis. Thus, PC modifies macrophage activity, but this effect depends on cell activation state, fatty acid saturation and esterification to PC molecule and PC concentration. Taken together, these results indicate that the fatty acid moiety of PC modulates macrophage activity and, consequently, is likely to affect immune system regulation in vivo.