New species of Ischnocnema (Anura, Brachycephalidae) from the atlantic rainforest of the state of Espirito Santo, Brazil.

We describe a new species of Ischnocnema from Santa Teresa, in the Atlantic Rainforest of the State of Espirito Santo, Brazil. Ischnocnema abdita sp. nov. is a member of the Ischnocnema lactea species series; it differs from other Ischnocnema mainly by presenting small size (adult male SVL 15.7 [plus or minus] 0.49 mm; range 15.0-16.8 mm); small discs on fingers I and II, and elliptical, wider than long, moderate sized discs on fingers III and IV; and finger I slightly smaller than finger II. The new species closely resembles Ischnocnema bolbodactyla, differing from it by presenting upper and lower margins of iris red in life; inguinal region with many yellow blotches in life; posterior region of belly and ventral surfaces of thighs, shanks, and feet roughly marbled with large pale yellow blotches; and hidden dorsal area of thighs dark brown with few large pale yellow blotches.

Physiological importance of the coronary arterial blood supply to the rattlesnake heart

The reptilian heart consists of a thick inner spongy myocardium that derives its oxygen and nutrient supply directly from the blood within the ventricular cavity, which is surrounded by a thin outer compact layer supplied by coronary arteries. The functional importance of these coronary arteries remains unknown. In the present study we investigate the effects of permanent coronary artery occlusion in the South American rattlesnake (Crotalus durissus) on the ability to maintain heart rate and blood pressure at rest and during short term activity. We used colored silicone rubber (Microfil) to identify the coronary artery distribution and interarterial anastomoses. The coronary circulation was occluded and the snakes were then kept for 4 days at 30 degrees C. Microfil injections verified that virtually all coronary arteries had successfully been occluded, but also made visible an extensive coronary supply to the outer compact layer in untreated snakes. Electrocardiogram (ECG), blood pressure (P(sys)) and heart rate (f(H)) were measured at rest and during enforced activity at day 1 and 4. Four days after occlusion of the coronary circulation, the snakes could still maintain a P(sys) and f(H) of 5.2 +/- 0.2 kPa and 58.2 +/- 2.2 beats min(-1), respectively, during activity and the ECG was not affected. This was not different from sham-operated snakes. Thus, while the outer compact layer of the rattlesnake heart clearly has an extensive coronary supply, rattlesnakes sustain a high blood pressure and heart rate during activity without coronary artery blood supply.

Niche occupancy and the relative role of micro-habitat and diet in resource partitioning among pond dwelling tadpoles.

We determined microhabitat and diet niche for tadpoles from two ponds in an agricultural landscape. Additionally, we verified the intraspecific variation in resource use, and if diet and microhabitat use were correlated. Tadpoles found in the two ponds differed in microhabitat use, because in the larger pond they explored deeper places far from the margin. There were three groups with high microhabitat niche overlap. In both ponds, plant cover was the best descriptor to explain interspecific variation in microhabitat use. Tadpoles of all species ingested mainly Bacillariophyceae and Trachellomonas however the diet differed intraspecifically in the species from the two ponds. Ten items in the temporary pond and 15 items in the permanent one were ingested by all species; however, the relative abundance of each item differed. Diet similarity was not correlated to similarity in microhabitat use. In this study, diet was as important as microhabitat use to explain resource partitioning.

Initial structural analysis of an alpha(4)beta(4) C-type lectin from the venom of Crotalus durissus terrificus

Convulxin, an alphabeta C-type lectin, is a potent platelet activator isolated from the venom of the South American rattlesnake Crotalus durissus terrificus. It is a 26.5 kDa alphabeta heterodimer consisting of two homologous disulfide-linked chains. The crystals belong to space group I4, with unit-cell parameters a = b = 131.61, c = 121.85 Angstrom, and diffraction data were collected to 2.7 Angstrom. The structure was solved by molecular replacement and the asymmetric unit contains two alphabeta heterodimers, each of which forms a disulfide-linked cyclic alpha(4)beta(4) tetramer in the unit cell. These alpha(4)beta(4) tetramers are stacked to form a large solvent channel.

Crystal structure of the platelet activator convulxin, a disulfide-linked alpha(4)beta(4) cyclic tetramer from the venom of Crotalus durissus terrificus

Convulxin (CVX), a C-type lectin, isolated from the venom of the South American rattlesnake Crotalus durissus terrificus, causes cardiovascular and respiratory disturbances and is a potent platelet activator which hinds to platelet glycoprotein GPVI. The structure of CVX has been solved at 2.4 Angstrom resolution to a crystallographic residual of 18.6% (R-free =26.4%). CVX is a disulfide linked heterodimer consisting of homologous alpha and beta chains. The heterodimers are additionally linked by disulfide bridges to form cyclic alpha(4)beta(4)heterotetramers. These domains exhibit significant homology to the carbohydrate-binding domains of C-type lectins, to the factor IX-binding protein (IX-bp), and to flavocetin-A (Fl-A) but sequence and Structural differences are observed in both the domains in the putative Ca2+ and carbohydrate binding regions. (C) 2003 Elsevier B.V. All rights reserved.

Crotacetin, a novel snake venom C-type lectin homolog of convulxin, exhibits an unpredictable antimicrobial activity

Snake venom (sv) C-type lectins encompass a group of hemorrhagic toxins that are capable of interfering with blood stasis. A very well-studied svC-type lectin is the heterodimeric toxin, convulxin (CVX), from the venom of South American rattlesnake Crotalus durissus terrificus. CVX is able to activate platelets and induce their aggregation by acting via p62/GPVI collagen receptor. By using polymerase chain reaction homology screening, we have cloned several cDNA precursors of CVX subunit homologs. One of them, named crotacetin (CTC) beta-subunit, predicts a polypeptide with a topology very similar to the tridimensional conformations of other subunits of CVX-like snake toxins, as determined by computational analysis. Using gel permeation and reverse-phase high-performance liquid chromatography, CTC was purified from C. durissus venoms. CTC can be isolated from the venom of several C. durissus subspecies, but its quantitative predominance is in the venom of C. durissus cascavella. Functional analysis indicates that CTC induces platelet aggregation, and, importantly, exhibits an antimicrobial activity against Gram-positive and -negative bacteria, comparable with CVX.

Crystallization and preliminary X-ray crystallographic analysis of the heterodimeric crotoxin complex and the isolated subunits crotapotin and phospholipase A(2)

Crotoxin, a potent neurotoxin from the venom of the South American rattlesnake Crotalus durissus terrificus, exists as a heterodimer formed between a phospholipase A(2) and a catalytically inactive acidic phospholipase A(2) analogue (crotapotin). Large single crystals of the crotoxin complex and of the isolated subunits have been obtained. The crotoxin complex crystal belongs to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 38.2, b = 68.7, c = 84.2 angstrom, and diffracted to 1.75 angstrom resolution. The crystal of the phospholipase A(2) domain belongs to the hexagonal space group P6(1)22 (or its enantiomorph P6(5)22), with unit-cell parameters a = b = 38.7, c = 286.7 angstrom, and diffracted to 2.6 angstrom resolution. The crotapotin crystal diffracted to 2.3 angstrom resolution; however, the highly diffuse diffraction pattern did not permit unambiguous assignment of the unit-cell parameters.

Snake Venomic of Crotalus durissus terrificus-Correlation with Pharmacological Activities

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

Preliminary functional characterization, cloning and primary sequence of Fastuosain, a cysteine peptidase isolated from fruits of Bromelia fastuosa

The present work reports the characterization of Fastuosain, a novel cysteine protease of 25kDa, purified from the unripe fruits of Bromelia fastuosa, a wild South American Bromeliaceae. Proteolytic activity, measured using casein and synthetic substrates, was dependent on the presence of thiol reagents, having maximum activity at pH 7.0. The present work reports cDNA cloning of Fastuosain; cDNA was amplified by PCR using specific primers. The product was 1096pb long. Mature fastuosain has 217 residues, and with the proregion has a total length of 324 residues. Its primary sequence showed high homology with ananain(74%), stem bromelain (66%) and papain (44%).

Deglaciation sequences in the Permo-Carboniferous Itarare Group, Parana Basin, southern Brazil

The Late Carboniferous-Early Permian Itarare Group is a thick glacial unit of the Parana Basin. Five unconformity-bounded sequences have been defined in the eastern outcrop belt and recognized in well logs along 400 km across the central portion of the basin. Deglaciation sequences are present in the whole succession and represent the bulk of the stratigraphic record. The fining-upward vertical facies succession is characteristic of a retrogradational stacking pattern and corresponds to the stratigraphic record of major ice-retreat phases. Laterally discontinuous subglacial tillites and boulder beds occur at the base of the sequences. When these subglacial facies are absent, deglaciation sequences lie directly on the basal disconformities. Commonly present in the lowermost portions of the deglaciation sequences, polymictic conglomerates and cross-bedded sandstones are generated in subaqueous proximal outwash fans in front of retreating glaciers. The overlying assemblage of diamictites, parallel-bedded and rippled sandstones, and Bouma-like facies sequences are interpreted as deposits of distal outwash fan lobes. The tops of the deglaciation sequences are positioned in clay-rich marine horizons that show little (fine-laminated facies with dropstones) or no evidence of glacial influence on the deposition and likely represent periods of maximum ice retreat. (c) 2006 Elsevier Ltd. All rights reserved.

Sedimentation of the Cretaceous Bauru Group in São Paulo, Parana Basin, Brazil

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

Implications of groundwater weathered profile interactions to the mobilization of radionuclides

This study reports the nature and extent of open-system interaction between groundwater and a weathered profile developed in the high grade thorium and REE ore body in Morro do Ferro, Pocos de Caldas plateau. The radioelement mobility in the shallow oxidizing environment was considered by using chemical data in conjunction with U-234/U-238, Ra-226/Th-230, Th-230/U-234, Th-228/Th-232, Ra-228/Th-232 and Th-230/Ra-228 activity ratios (AR's) for borehole spoil and groundwater samples.Recharging groundwater from the studied borehole has low salinity values, with total dissolved solids content of 14.7 mg/l and total ionic strength of 0.00018. The ratio of the weight of dissolved radioelement per unit volume of solution to the weight of radioelement in solid phase per unit weight of solid phase showed that the radioelement solubility in the studied waters varied according to the following order: radium> uranium >thorium.U-234/U-238 AR's less than 1 were measured in solid phase and can justify the enhancement of U-234 in solution. Ra-226/Th-230 AR's greater than 1 and Th-230/U-234 AR'S less than 1 were evaluated between 20 and 27 m in depth, where a 2.1-m thick magnetite dike was intersected. These ratios could be justified by deposition of U and Ra associated with Fe-Mn oxides and kaolinite, where mineral saturation indices evaluated from the available data confirm this possibility. Covariations among disequilibria involving Th-228/(228) Ra and Ra-228/Th-232 AR's showed the preferential removal of Ra-228 relative to Tn isotopes, Th-228 and Th-232. The recent deposition of radium within the timescale of at least the last 35 years also is suggested. (C) 1998 Elsevier B.V. Ltd. All rights reserved.

Some possible evolutionary scenarios suggested by Cl-36 measurements in Guarani aquifer groundwaters

The Guarani aquifer underlies 1.2 M km(2) in the Parana sedimentary basin of South America and is an important source of water for industry, agriculture, and domestic supplies. To determine the sustainability of this aquifer we need to understand the dynamics of the groundwater system. This paper describes the first Cl-36 measurements on aquifer groundwaters and some measurements on South American rainwaters, thought to be indicative of the recharge water. The results are compared to previous work in the region, including other radioisotope analyses. A simple model is developed, incorporating radioactive decay, allowing scenarios to be developed for mixing different waters at different mixing rates. Thus, mixing scenarios consistent with other hydrogeological and hydrogeochemical data could be assessed. A model that mixes fresh recharging waters with formational waters, that contain elevated chloride levels, but low (in situ) Cl-36 levels, can explain most of the results presented here. The expectation that rainwater samples would provide a good end-member for modelling recharge proved problematic, however. As a consequence, it is suggested that either: the recharge waters are not sourced from the same locations as the rains; that the current rainfall and fallout conditions were significantly different in the past; or that the low levels of chloride in rainfall may have allowed some contamination of the samples by old (Cl-36-free) chloride during the recharge process. (c) 2008 Elsevier Ltd. All rights reserved.

Dissolved Po-210 and Pb-210 in Guarani aquifer groundwater, Brazil

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

Avaliação de mutagenicidade e antimutagenicidade de diferentes frações de Pterogyne nitens (Leguminosae), utilizando ensaio de micronúcleo em Tradescantia pallida

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