Molecular analysis reveals hidden diversity in <Emphasis Type="Italic">Zungaro</Emphasis> (Siluriformes: Pimelodidade): a genus of giant South American catfish

The genus Zungaro contains some of the largest catfish in South America. Two valid species are currently recognized: Zungaro jahu, inhabiting the Paraná and Paraguay basins, and Zungaro zungaro, occurring in the Amazonas and Orinoco basins. Analysing Zungaro specimens from the Amazonas, Orinoco, Paraguay and Paraná basins, based on the sequencing of COI and D-loop, we found at least three MOTUs, indicating the existence of hidden diversity within this fish group. Considering the ecological and economic values of this fish, our results are surely welcomed for its conservation, disclosing new findings on its diversity and pointing out the necessity for a detailed taxonomic revision.     

Population genetic structure of the Amazonian black flannelmouth characin (Characiformes,Prochilodontidae: <Emphasis Type="Italic">Prochilodus nigricans</Emphasis> Spix &amp; Agassiz, 1829): contemporary and historical gene flow of a migratory and abundant fishery species

In the present study, sequences of the mtDNA control region (834 bp) were analyzed from 337 specimens of Prochilodus nigricans from sites along the main channel of the Amazonas River and three major tributaries, Madeira, Purus, and Juruá. The results of the analysis of molecular variance revealed that a large part of the genetic variation occurred within the populations analyzed (~85 %). Analysis with SAMOVA and Barriers suggested that the upper Madeira River and Purus Rivers had diverged genetically from the other samples, indicating restricted gene flow among these areas, while sites within the remaining range exhibited relatively little population structure. The high degree of structuring observed in the Madeira River basin population may be attributed to the presence of rapids along its upper course, while the genetic divergence found in the upper Purus River suggests historical connection between the upper Purus and upper Madeira Rivers followed by slow genetic drift due to large effective population sizes. However, given the life history and hypothesized evolutionary strategy of this species, we urge caution in interpreting that this targeted species is not at risk of overexploitation due to contemporary abundance. In order to preserve genetic diversity, we recommend enforcement of management regimes for regional stocks.     

Selenium Status in Patients with Turner Syndrome: a Biochemical Assessment Related with Body Composition

Studies about selenium status in patients with Turner syndrome (TS) are non-existent in the literature. The aim of this study was to evaluate selenium status in patients with TS, while considering the different ages of the studied population and the relation with body composition. In total, 33 patients with TS were evaluated and grouped according to their developmental stages (children, adolescents, and adults). Selenium concentrations in their plasma, erythrocytes, urine, and nails were determined by using hydride generation atomic absorption spectrometry and erythrocyte glutathione peroxidase activity were measured by using Randox commercial kits. Additionally, height, weight, body fat percentage, waist circumference, and waist-height ratio were measured to characterize the patients. No differences in the selenium concentrations in the plasma, erythrocyte, urine, and nails or in the glutathione peroxidase activity were observed among the age groups (p > 0.05). The evaluated selenium levels were less than the established normal ones. The patients with larger waist circumference, body fat percentage, body mass index, and waist-height ratio showed lower glutathione peroxidase enzyme activity (p = 0.023). The present study shows that most patients with TS are deficient in selenium and that those with a greater accumulation of body fat have a lower GPx activity.     

Patterns of diversification in the discus fishes (Symphysodon spp. Cichlidae) of the Amazon basin

We carried out a phylogeograhic and population genetic analysis of fishes of the taxonomically contentious genus Symphysodon from the Amazon basin in order to test hypotheses of relationships among taxonomic units, and potential processes driving diversification within this genus. We sampled 334 individuals of the genus Symphysodon from 24 localities that span the complete geographic distribution of this genus. The sampling scheme included all known phenotypic groups, species and subspecies. Analyses were based on 474 bp of the mitochondrial control region and 1443 bp of the exon 3 of RAG1 gene. We observed 102 mtDNA haplotypes defined by 89 segregating sites, and 5 nuDNA alleles defined by three segregating sites. Maximum-likelihood, Bayesian-inference and statistical parsimony analyses revealed three well defined monophyletic groups. These clades corresponded to the 'green' and 'blue' groups of Symphysodon aequifasciatus, and to a previously morphologically unrecognized clade from the Xingu River drainage. These three clades were nested within a paraphyletic assemblage consisting of the 'brown' group of S. aequifasciatus and of both described subspecies of S. discus, the 'Heckel' and the 'abacaxi' discus. Nuclear allele sharing was observed among groups, but there were significant differences in frequencies. We inferred several processes including past fragmentation among groups, and restricted gene flow with isolation by distance within the paraphyletic 'brown+Heckel+abacaxi' groups, and suggest that differences among the 'blue', 'Heckel' and 'brown' groups are potentially maintained by differences in water chemistry preferences. We further inferred colonization of the western Amazon basin by an ancestor of the 'green' clade. The 'green' group was the only group with a pattern of haplotype distribution consistent of a demographic expansion, and the divergence of this clade from other groups of discus was consistent with recent geologic evidence on the breach of the Purus Arch which separates western Amazon from eastern Amazon. We further hypothesized that the differentiation of the 'Xingu' clade could be due to vicariance events resulting from Pleistocene sea level, and thus Amazon River level fluctuations. We discuss the bearings of our results on the current taxonomy of this group, and on the biological reality of the different forms, subspecies and species of Symphysodon concluding that we are probably observing a process of diversification, and therefore taxonomy will remain contentious.     

Characterization of microsatellites from cacao–<Emphasis Type="Italic">Moniliophthora perniciosa</Emphasis> interaction expressed sequence tags

Theobroma cacao L.–Moniliophthora perniciosa expressed sequence tags (ESTs) were converted into useful satellite markers for population analysis and genetic mapping. Forty-nine flanking primer pairs from TSH1188 (a resistant genotype) and Catongo (a susceptible genotype) ESTs were designed and screened for polymorphism analysis. Eleven were polymorphic, with an average of 3.81 alleles per locus and a total of 42 alleles. The satellite markers were tested on 21 cacao accessions and two bulked DNAs generated from 6 resistant and 6 susceptible plants from a segregating F₂ (SCA6 × ICS1) population for witches’ broom resistance. These results show that EST-derived microsatellites (short sequence repeats, SSRs) in Theobroma cacao have many potential applications in linkage mapping and the planning of crosses.     

Characterization of nucleoside triphosphate diphosphohydrolase activity in Trichomonas gallinae and the influence of penicillin and streptomycin in extracellular nucleotide hydrolysis

Here we described an nucleoside triphosphate diphosphohydrolase (NTPDase) activity in living trophozoites of Trichomonas gallinae. The enzyme hydrolyzes a variety of purine and pyrimidine nucleoside di- and triphosphates in an optimum pH range of 6.0-8.0. This enzyme activity was activated by high concentrations of divalent cations, such as calcium and magnesium. Contaminant activities were ruled out because the enzyme was not inhibited by classical inhibitors of ATPases (ouabain, 5.0 mM sodium azide, oligomycin) and alkaline phosphatases (levamisole). A significant inhibition of ATP hydrolysis (38%) was observed in the presence of 20 mM sodium azide. Sodium orthovanadate inhibited ATP and ADP hydrolysis (24% and 78%), respectively. The apparent K(M) (Michaelis constant) values were 667.62+/-13 microM for ATP and 125+/-5.3 microM for ADP. V(max) (maximum velocity) values were 0.44+/-0.007 nmol Pi min(-1) per 10(6) trichomonads and 0.91+/-0.12 nmol Pi min(-1) per 10(6) trichomonads for ATP and ADP, respectively. Moreover, we showed a marked decrease in ATP, ADP and AMP hydrolysis when the parasites were grown in the presence of penicillin and streptomycin. The existence of an NTPDase activity in T. gallinae may be involved in pathogenicity, protecting the parasite from the cytolytic effects of the extracellular nucleotides.     

Poly(3-hydroxybutyrate)/chitosan/ketoprofen or piroxicam composite microparticles: Preparation and controlled drug release evaluation

Poly(3-hydroxybutyrate)/chitosan/piroxicam or ketoprofen composite microparticles were prepared by the solid-in-water-in-oil emulsion-solvent evaporation technique with the aim of reducing the burst effect and controlling the drug release. Reservoir-type microparticles, composed of poly(3-hydroxybutyrate) microspheres embedded in a chitosan matrix were prepared. The size and morphological characteristics of the composite microparticles were evaluated in relation to the chitosan concentration and cross-linking with glutaraldehyde. Reservoir-type composite microparticles were obtained using 2.0% and 3.0% w/v chitosan solutions. A significant reduction in the burst effect and prolonged drug release were observed, particularly when higher chitosan and glutaraldehyde concentrations were used.     

Structure and Mode of Action of Microplusin, a Copper II-chelating Antimicrobial Peptide from the Cattle Tick Rhipicephalus (Boophilus) microplus

Microplusin, a Rhipicephalus (Boophilus) microplus antimicrobial peptide (AMP) is the first fully characterized member of a new family of cysteine-rich AMPs with histidine-rich regions at the N and C termini. In the tick, microplusin belongs to the arsenal of innate defense molecules active against bacteria and fungi. Here we describe the NMR solution structure of microplusin and demonstrate that the protein binds copper II and iron II. Structured as a single α-helical globular domain, microplusin consists of five α-helices: α1 (residues Gly-9 to Arg-21), α2 (residues Glu-27 to Asn-40), α3 (residues Arg-44 to Thr-54), α4 (residues Leu-57 to Tyr-64), and α5 (residues Asn-67 to Cys-80). The N and C termini are disordered. This structure is unlike any other AMP structures described to date. We also used NMR spectroscopy to map the copper binding region on microplusin. Finally, using the Gram-positive bacteria Micrococcus luteus as a model, we studied of mode of action of microplusin. Microplusin has a bacteriostatic effect and does not permeabilize the bacterial membrane. Because microplusin binds metals, we tested whether this was related to its antimicrobial activity. We found that the bacteriostatic effect of microplusin was fully reversed by supplementation of culture media with copper II but not iron II. We also demonstrated that microplusin affects M. luteus respiration, a copper-dependent process. Thus, we conclude that the antibacterial effect of microplusin is due to its ability to bind and sequester copper II.     

Mitotic instability in resynthesized and natural polyploids of the genus Arabidopsis (Brassicaceae)

Allopolyploids contain complete sets of chromosomes from two or more different progenitor species. Because allopolyploid hybridization can lead to speciation, allopolyploidy is an important mechanism in evolution. Meiotic instability in early-generation allopolyploids contributes to high lethality, but less is known about mitotic fidelity in allopolyploids. We compared mitotic stability in resynthesized Arabidopsis suecica-like neoallopolyploids with that in 13 natural lines of A. suecica (2n = 4x = 26). We used fluorescent in situ hybridization to distinguish the chromosomal contribution of each progenitor, A. thaliana (2n = 2x =10) and A. arenosa (2n = 4x = 32). Surprisingly, cells of the paternal parent A. arenosa had substantial aneuploidy, while cells of the maternal parent A. thaliana were more stable. Both natural and resynthesized allopolyploids had low to intermediate levels of aneuploidy. Our data suggest that polyploidy in Arabidopsis is correlated with aneuploidy, but varies in frequency by species. The chromosomal composition in aneuploid cells within individuals was variable, suggesting somatic mosaicisms of cell lineages, rather than the formation of distinct, stable cytotypes. Our results suggest that somatic aneuploidy can be tolerated in Arabidopsis polyploids, but there is no evidence that this type of aneuploidy leads to stable novel cytotypes.     

Immunization protected well nourished mice but not undernourished ones from lung injury in Methicillin-resistant Staphylococcus aureus (MRSA) infection

Background  

Staphylococcus aureus methicillin-resistant (MRSA) has been frequently isolated from endotracheal and lung puncture aspirates in malnourished children with pneumonia. In this work we evaluated the susceptibility of undernourished BALB/c mice and its ability to mount a protective immunity against MRSA with emphasis on the lung involvement.