Guiana dolphins, Sotalia guianensis (Cetacea: Delphinidae), in the Paranaguá Estuarine Complex: insights on the use of area based on the photo-identification technique

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Structure of a lectin from Canavalia gladiata seeds: new structural insights for old molecules

Background: Lectins are mainly described as simple carbohydrate- binding proteins. Previous studies have tried to identify other binding sites, which possible recognize plant hormones, secondary metabolites, and isolated amino acid residues. We report the crystal structure of a lectin isolated from Canavalia gladiata seeds ( CGL), describing a new binding pocket, which may be related to pathogen resistance activity in ConA- like lectins; a site where a non- protein amino- acid, aaminobutyric acid ( Abu), is bound.Results: the overall structure of native CGL and complexed with alpha- methyl- mannoside and Abu have been refined at 2.3 angstrom and 2.31 angstrom resolution, respectively. Analysis of the electron density maps of the CGL structure shows clearly the presence of Abu, which was confirmed by mass spectrometry.Conclusion: the presence of Abu in a plant lectin structure strongly indicates the ability of lectins on carrying secondary metabolites. Comparison of the amino acids composing the site with other legume lectins revealed that this site is conserved, providing an evidence of the biological relevance of this site. This new action of lectins strengthens their role in defense mechanisms in plants.

New insights into the in vitro organogenesis process: the case of Passiflora

We present evidences that ultrastructural electron microscope findings are valuable ways to understand the in vitro regeneration process, in particular in the yellow passion fruit. Shoot-regeneration was induced in hypocotyl and leaf-derived explants using 4.44 mu M BAP, and the entire organogenic process was analyzed using conventional histology, scanning and transmission electronic microscopy. Both direct and indirect regeneration modes were observed in hypocotyl explants, but only direct regeneration occurred in leaf-derived cultures. In the direct pathway from both explant types, meristemoids developed into globular structures, here called protuberances. The peripheral meristematic layers of the protuberances displayed ultrastructural characteristics indicative of a high metabolic activity, and only these cells originated shoots and leaf primordia, the latter being frequent when leaf explants were used. Moreover, the peripheral cells of the protuberances derived from leaf explants lost adhesion during the culture, diminishing the regeneration rates. We recommend the use of hypocotyls as a source of explant to obtain shoots as well as a genetic transformation system for the yellow passion fruit. However, the direct pathway is preferred because a type of amitosis occurred in the peripheral cells of hypocotyl-derived calli, which has the potential to result in genetic instability of the regenerating plants/tissue.

Transcriptome profiling of Paracoccidioides brasiliensis yeast-phase cells recovered from infected mice brings new insights into fungal response upon host interaction

Paracoccidioides brasiliensis is a fungal human pathogen with a wide distribution in Latin America. It causes paracoccidioidomycosis, the most widespread systemic mycosis in Latin America. Although gene expression in P. brasiliensis had been studied, little is known about the genome sequences expressed by this species during the infection process. To better understand the infection process, 4934 expressed sequence tags (ESTs) derived from a non-normalized cDNA library from P. brasiliensis (isolate Pb01) yeast-phase cells recovered from the livers of infected mice were annotated and clustered to a UniGene (clusters containing sequences that represent a unique gene) set with 1602 members. A large-scale comparative analysis was performed between the UniGene sequences of P. brasiliensis yeast-phase cells recovered from infected mice and a database constructed with sequences of the yeast-phase and mycelium transcriptome (isolate Pb01) (https://dna.biomol.unb.br/Pb/), as well as with all public ESTs available at GenBank, including sequences of the P. brasiliensis yeast-phase transcriptome (isolate Pb18) (http:// www.ncbi.nlm.nih.gov/). The focus was on the overexpressed and novel genes. From the total, 3184 ESTs (64.53%) were also present in the previously described transcriptome of yeast-form and mycelium cells obtained from in vitro cultures (https://dna.biomol.unb.br/Pb/) and of those, 1172 ESTs (23.75% of the described sequences) represented transcripts overexpressed during the infection process. Comparative analysis identified 1750 ESTs (35.47% of the total), comprising 649 UniGene sequences representing novel transcripts of P. brasiliensis, not previously described for this isolate or for other isolates in public databases. KEGG pathway mapping showed that the novel and overexpressed transcripts represented standard metabolic pathways, including glycolysis, amino acid biosynthesis, lipid and sterol metabolism. The unique and divergent representation of transcripts in the cDNA library of yeast cells recovered from infected mice suggests differential gene expression in response to the host milieu.

Structural analysis of Canavalia maritima and Canavalia gladiata lectins complexed with different dimannosides: New insights into the understanding of the structure-biological activity relationship in legume lectins

Plant lectins, especially those purified from species of the Legummosae family, represent the best studied group of carbohydrate-binding proteins. The legume lectins from Diocleinae subtribe are highly similar proteins that present significant differences in the potency/ efficacy of their biological activities. The structural studies of the interactions between lectins and sugars may clarify the origin of the distinct biological activities observed in this high similar class of proteins. In this way, this work presents a crystallographic study of the ConM and CGL (agglutinins from Canavalia maritima and Canavalia gladiata, respectively) in the following complexes: ConM/ CGL:Man(alpha 1-2)Man(alpha 1-0)Me, ConM/CGL:Man(alpha 1-O)Man(alpha 1-O)Me and ConM/CGL:Man(alpha 1-4)Man(alpha 1-O)Me, which crystallized in different conditions and space group from the native proteins.The structures were solved by molecular replacement, presenting satisfactory values for R-factor and R-factor. Comparisons between ConM, CGL and ConA (Canavalia ensiformis lectin) binding mode with the dimannosides in subject, presented different interactions patterns, which may account for a structural explanation of the distincts biological properties observed in the lectins of Diocleinae subtribe. (C) 2007 Elsevier B.V. All rights reserved.

Intrageneric karyotypic divergence in Scythrophrys and new insights into the relationship with Paratelmatobius (Anura, Leptodactylidae)

Three populations of Scythrophrys from Brazil were studied cytogenetically. The karyotype found in Piraquara (Parana State) and Sao Bento do Sul (Santa Catarina State) (karyotype 1) differed from that of Rancho Queimado (Santa Catarina State) (karyotype 11) in NOR location and in the occurrence of some C-bands. In karyotype I, pair 5 contained NOR-bearing chromosomes, while in karyotype 11 the NOR occurred in pair 10. This difference probably accounts for the inverted classification of pairs 5 and 6 in these karyotypes, which appear to be homologous, as suggested by the presence of heterochromatic markers. These karyotypes also differed in the size of the non-centromeric C-bands in pairs 1 and 8. Comparison of the Scythrophrys karyotypes with those of the related genus Paratelmatobius revealed several similarities, especially between karyotype 11 of Scythrophrys and that of P. poecilogaster. Both karyotypes had a NOR in pair 10, so it was inferred to be present also in the common ancestor for these genera. The non-centromeric C-bands in pairs 7, 10, and in pair 5 of karyotype I and pair 6 of karyotype II of Scythrophrys are characters exclusively found in this genus.

Mechanistic insights into the reaction between VO2+ and propene based on a DFT study

Calculations based on density functional theory have been carried out to investigate the free energy profiles at singlet and triplet electronic states associated with the gas-phase ion/molecule reactions of VO2++ ((1)A(1)/(3)A) with propene. The complex potential energy Surfaces, including Six reaction pathways (three dehydrogenation and three oxygen transfer processes), have been explored and analyzed. Along dehydrogenation reactive channels, three final products can be obtained: V(OH)(2)(+) ((1)Sigma(+)/(3)Sigma(-)) and allene (path Dehl), being the most kinetically and thermodynamically favorable reaction pathway, V(OH)(2)(+) ((1)Sigma(+)/(3)Sigma(-)) and propyne (path Deh2),and VO2+ ((1)A(1)/(3)A) and H-2 plus allene (path Deh3). The oxyoenation processes can yield its final products Vo(+) ((1)Delta/(3)Sigma) and acetone (path Ox1), VO+ ((1)Delta/(3)Sigma 2) and propanaldehyde (path Ox2), and VO+ ((1)Delta/(3)Sigma) and H-2 and propenaldehyde (path Ox3). Both paths Deh1 and Deh2 are associated with two consecutive hydrogen transfer processes from carbon atoms of the propene fragment to vanadyl oxygen atoms, while in path Deh3 the second hydrogen migration takes place to the vanadiurn atorn followed by the formation ola hydrogen molecule. Both paths Ox1 and Ox2 comprise an intramolecular hydrogen transfer between the ethylenic moiety of the propene fragment, while two consecutive hydrogen transfer processes take place from the propene fragment to oxygen and vanadium atoms of the vanadyl moiety along path Ox3. Three crossing points between both electronic states take place along path Deh1 (CP-Deh1) and path Deh2 (CP-Deh2) and in the entrance channel of oxidation processes (CP-Ox). A comparison with previous works on related reactions VO2+ + C2H4, VO2 + C2H6, and VO2+ + C3H8 allows us to rationalize the different reactivity patterns.

Anisotropic growth of oxide nanocrystals: Insights into the rutile TiO2 phase

In this work, we report the synthesis of titanium oxide nanocrystals, especially the rutile TiO2 phase with nanorod morphology, by a method based on peroxotitanium complex decomposition. The results indicate that the anisotropic morphology reported for rutile TiO2 nanocrystals is related to the oriented attachment process. Despite the predominance of rutile nanocrystals at longer treatment times, the nanocrystals were obtained also in the anatase type, according to the degradation time adopted. XANES results evidenced the absence of structural correlation between the peroxytitanium complex and phase evolution, and the coexistence of the two phases strongly suggests a correlation of the oriented attachment mechanism and the rutile phase stabilization.

Insights into the concept of fish welfare

Fish welfare issues are predicated on understanding whether fish are sentient beings. Therefore, we analyzed the logic of the methodologies used for studying this attribute. We conclude that empirical science is unable to prove or to disprove that fish are sentient beings. Thus, we propose a combined ethical-scientific approach for considering fish as sentient beings. The most difficult ongoing question is to determine which conditions fish prefer. Approaches to assess fish preferences should be rigorously and cautiously employed. In light of these considerations, attempts to establish physiological standards for fish welfare are discouraged, and a preference-based definition of fish welfare is proposed.

The relationship between diet quality and basal metabolic rate in endotherms: Insights from intraspecific analysis

In this article, we review intraspecific studies of basal metabolic rate (BMR) that address the correlation between diet quality and BMR. The food-habit hypothesis stands as one of the most striking and often-mentioned interspecific patterns to emerge from studies of endothermic energetics. Our main emphasis is the explicit empirical comparison of predictions derived from interspecific studies with data gathered from within-species studies in order to explore the mechanisms and functional significance of the putative adaptive responses encapsulated by the food-habit hypothesis. We suggest that, in addition to concentrating on the relationship among diet quality, internal morphology, and BMR, new studies should also attempt to unravel alternative mechanisms that shape the interaction between diet and BMR, such as enzymatic plasticity, and the use of energy-saving mechanisms, such as torpor. Another avenue for future study is the measurement of the effects of diet quality on other components of the energy budget, such as maximum thermogenic and sustainable metabolic rates. It is possible that the effects of diet quality operate on such components rather than directly on BMR, which might then push or pull along changes in these traits. Results from intraspecific studies suggest that the factors responsible for the association between diet and BMR at an ecological timescale might not be the same as those that promoted the evolution of this correlation. Further analyses should consider how much of a role the proximate and ultimate processes have played in the evolution of BMR.

The Serido Group of NE Brazil, a late Neoproterozoic pre- to syn-collisional basin in West Gondwana: insights from SHRIMP U-Pb detrital zircon ages and Sm-Nd crustal residence (T-DM) ages

The Serido Group is a deformed and metamorphosed metasedimentary sequence that overlies early Paleoproterozoic to Archean basement of the Rio Grande do Norte domain in the Borborema Province of NE Brazil. The age of the Serido Group has been disputed over the past two decades, with preferred sedimentation ages being either Paleoproterozoic or Neoproterozoic. Most samples of the Serido Formation, the upper part of the Serido Group, have Sm-Nd T-DM ages between 1200 and 1600 Ma. Most samples of the Jucurutu Formation, the lower part of the Serido Group, have T-DM ages ranging from 1500 to 1600 Ma; some basal units have T-DM ages as old as 2600 Ma, reflecting proximal basement. Thus, based on Sm-Nd data, most, if not all, of the Serido Group was deposited after 1600 Ma and upper parts must be younger than 1200 Ma.Cathodoluminescence photos of detrital zircons show very small to no overgrowths produced during ca. 600 Ma Brasiliano deformation and metamorphism, so that SHRIMP and isotope dilution U-Pb ages must represent crystallization ages of the detrital zircons. Zircons from meta-arkose near the base of the Jucurutu Formation yield two groups of ages: ca. 2200 Ma and ca. 1800 Ma. In contrast, zircons from a metasedimentary gneiss higher in the Jucurutu Formation yield much younger ages, with clusters at ca. 1000 Ma and ca. 650 Ma. Zircons from metasedimentary and metatuffaceous units in the Serido Formation also yield ages primarily between 1000 and 650 Ma, with clusters at 950-1000, 800, 750, and 650 Ma. Thus, most, if not all, of the Serido Group must be younger than 650 Ma. Because these units were deformed and metamorphosed in the ca. 600 Ma Brasiliano fold belt during assembly of West Gondwana, deposition probably occurred ca. 610-650 Ma, soon after crystallization of the youngest population of zircons and before or during the onset of Brasiliano deformation.The Serido Group was deposited upon Paleoproterozoic basement in a basin receiving detritus from a variety of sources. The Jucurutu Formation includes some basal volcanic rocks and initially received detritus from proximal 2.2-2.0 Ga (Transamazonian) to late Paleoproterozoic (1.8-1.7 Ga) basement. Provenance for the upper Jucurutu Formation and all of the Serido Formation was dominated by more distal and younger sources ranging in age from 1000 to 650 Ma. We suggest that the Serido basin may have developed as the result of late Neoproterozoic extension of a pre-existing continental basement, with formation of small marine basins that were largely floored by cratonic basement (subjacent oceanic crust has not yet been found). Immature sediment was initially derived from surrounding land; as the basin evolved much of the detritus probably came from highlands to the south (present coordinates). Alternatively, if the Patos shear zone is a major terrane boundary, the basin may have formed as an early collisional foredeep associated with south-dipping subduction. In any case, within 30 million years the region was compressed, deformed, and metamorphosed during final assembly of West Gondwana and formation of the Brasiliano-Pan African fold belts. (C) 2003 Elsevier B.V. All rights reserved.

Comparative genomics of two Leptospira interrogans serovars reveals novel insights into physiology and pathogenesis

Leptospira species colonize a significant proportion of rodent populations worldwide and produce life-threatening infections in accidental hosts, including humans. Complete genome sequencing of Leptospira interrogans serovar Copenhageni and comparative analysis with the available Leptospira interrogans serovar Lai genome reveal that despite overall genetic similarity there are significant structural differences, including a large chromosomal inversion and extensive variation in the number and distribution of insertion sequence elements. Genome sequence analysis elucidates many of the novel aspects of leptospiral physiology relating to energy metabolism, oxygen tolerance, two-component signal transduction systems, and mechanisms of pathogenesis. A broad array of transcriptional regulation proteins and two new families of afimbrial adhesins which contribute to host tissue colonization in the early steps of infection were identified. Differences in genes involved in the biosynthesis of lipopolysaccharide 0 side chains between the Copenhageni and Lai serovars were identified, offering an important starting point for the elucidation of the organism's complex polysaccharide surface antigens. Differences in adhesins and in lipopolysaccharide might be associated with the adaptation of serovars Copenhageni and Lai to different animal hosts. Hundreds of genes encoding surface-exposed lipoproteins and transmembrane outer membrane proteins were identified as candidates for development of vaccines for the prevention of leptospirosis.

Experimental Basis and New Insights for Cell Therapy in Chronic Obstructive Pulmonary Disease

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Proteome analysis of snake venom toxins: pharmacological insights

Snake venoms are an extremely rich source of pharmacologically active proteins with a considerable clinical and medical potential. To date, this potential has not been fully explored, mainly because of our incomplete knowledge of the venom proteome and the pharmacological properties of its components, in particular those devoid of enzymatic activity. This review summarizes the latest achievements in the determination of snake venom proteome, based primarily on the development of new strategies and techniques. Detailed knowledge of the venom toxin composition and biological properties of the protein constituents should provide the scaffold for the design of new more effective drugs for the treatment of the hemostatic system and heart disorders, inflammation, cancer and consequences of snake bites, as well as new tools for clinical diagnostic and assays of hemostatic parameters.