975 resultados para Bothrops jararacussu snake venom


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Secretory phospholipases A(2) (sPLA(2)) exert proinflammatory actions through lipid mediators. These enzymes have been found to be elevated in many inflammatory disorders such as rheumatoid arthritis, sepsis, and atherosclerosis. The aim of this study was to evaluate the effect of harpalycin 2 (Har2), an isoflavone isolated from Harpalyce brasiliana Benth., in the enzymatic, edematogenic, and myotoxic activities of sPLA2 from Bothrops pirajai, Crotalus durissus terrificus, Apis mellifera, and Naja naja venoms. Har2 inhibits all sPLA(2) tested. PrTX-III (B. pirajai venom) was inhibited at about 58.7%, Cdt F15 (C. d. terrificus venom) at 78.8%, Apis (from bee venom) at 87.7%, and Naja (N. naja venom) at 88.1%. Edema induced by exogenous sPLA(2) administration performed in mice paws showed significant inhibition by Har2 at the initial step. In addition, Har2 also inhibited the myotoxic activity of these sPLA(2)s. In order to understand how Har2 interacts with these enzymes, docking calculations were made, indicating that the residues His48 and Asp49 in the active site of these enzymes interacted powerfully with Har2 through hydrogen bonds. These data pointed to a possible anti-inflammatory activity of Har2 through sPLA(2) inhibition.

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Tese de Doutoramento em Ciências Veterinárias na Especialidade de Ciências Biológicas e Biomédicas

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Mastoparans are tetradecapeptides found to be the major component of vespid venoms. A mastoparan toxin isolated from the venom of Anterhynchium flavomarginatum micado has been crystallized and X-ray diffraction data collected to 2.7 Angstrom resolution using a synchrotron-radiation source. Crystals were determined to belong to the space group P6(2)22 (P6(4)22). This is the first mastoparan to be crystallized and will provide further insights into the conformational significance of mastoparan toxins with respect to their potency and activity in G-protein regulation.

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Mastoparans are tetradecapeptides found to be the major component of vespid venoms. These peptides present a wide spectrum of biological activities, such as mast cell degranulation, hemolytic activity and also reveals antimicrobial activity. A mastoparan toxin isolated from the venom of Anterhynchium flavomarginatum micado has been crystallized. At room temperature these crystals diffracted to 2.8 Angstrom resolution. However, upon cooling to cryogenic temperature around 85 K, the original resolution limit could be improved to 2.0 Angstrom. Crystals were determined to belong to the space group P3(1) (P3(2)). This is the first mastoparan to be crystallized and it will provide further insights in the conformational significance of mastoparan toxins, with respect to their potency and activity in G protein regulation. (C) 3001 Elsevier B.V. B.V. All rights reserved.

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The performance of iris recognition systems is significantly affected by the segmentation accuracy, especially in non- ideal iris images. This paper proposes an improved method to localise non-circular iris images quickly and accurately. Shrinking and expanding active contour methods are consolidated when localising inner and outer iris boundaries. First, the pupil region is roughly estimated based on histogram thresholding and morphological operations. There- after, a shrinking active contour model is used to precisely locate the inner iris boundary. Finally, the estimated inner iris boundary is used as an initial contour for an expanding active contour scheme to find the outer iris boundary. The proposed scheme is robust in finding exact the iris boundaries of non-circular and off-angle irises. In addition, occlusions of the iris images from eyelids and eyelashes are automatically excluded from the detected iris region. Experimental results on CASIA v3.0 iris databases indicate the accuracy of proposed technique.

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Peptidases are ubiquitous enzymes involved in diverse biological processes. Fragments from bioactive peptides have been found in skin secretions from frogs, and their presence suggests processing by peptidases. Thus, the aim of this work was to characterize the peptidase activity present in the skin secretion of Leptodactylus labyrinthicus. Zymography revealed the presence of three bands of gelatinase activity of approximately 60 kDa, 66 kDa, and 80 kDa, which the first two were calcium-dependent. These three bands were inhibited either by ethylenediaminetetraacetic acid (EDTA) and phenathroline; thus, they were characterized as metallopeptidases. Furthermore, the proteolytic enzymes identified were active only at pH 6.0–10.0, and their activity increased in the presence of CHAPS or NaCl. Experiments with fluorogenic substrates incubated with skin secretions identified aminopeptidase activity, with cleavage after leucine, proline, and alanine residues. This activity was directly proportional to the protein concentration, and it was inhibited in the presence of metallo and serine peptidase inhibitors. Besides, the optimal pH for substrate cleavage was determined to be 7.0–8.0. The results of the in gel activity assay showed that all substrates were hydrolyzed by a 45 kDa peptidase. Gly-Pro-AMC was also cleaved by a peptidase greater than 97 kDa. The data suggest the presence of dipeptidyl peptidases (DPPs) and metallopeptidases; however, further research is necessary. In conclusion, our work will help to elucidate the implication of these enzymatic activities in the processing of the bioactive peptides present in frog venom, expanding the knowledge of amphibian biology.

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The influence of the membrane active peptides, Tat44–57 (activator in HIV-1) and melittin (active content of bee venom), on self-assembled monolayers of 6-mercaptohexanoic acid (MHA) on gold electrodes has been studied with scanning electrochemical microscopy (SECM). It was found that MHA, when deprotonated at physiological pH, significantly affected the relative rates of electron transfer between the [Fe(CN)6]4− solution based mediator and the underlying gold electrode, predominantly by the electrostatic interaction between the mediator and MHA. Upon the introduction of Tat44–57 ormelittin to the electrolyte, the relative rate of electron transfer through the MHA layer could be increased or decreased depending on the mediator used. However, in all cases it was found that these peptides have the ability to be incorporated into synthetic SAMs, which has implications for future electrochemical studies carried out using cell mimicking membranes immobilised on such layers.

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The 12.7-10.5 Ma Cougar Point Tuff in southern Idaho, USA, consists of 10 large-volume (>10²-10³ km³ each), high-temperature (800-1000 °C), rhyolitic ash-flow tuffs erupted from the Bruneau-Jarbidge volcanic center of the Yellowstone hotspot. These tuffs provide evidence for compositional and thermal zonation in pre-eruptive rhyolite magma, and suggest the presence of a long-lived reservoir that was tapped by numerous large explosive eruptions. Pyroxene compositions exhibit discrete compositional modes with respect to Fe and Mg that define a linear spectrum punctuated by conspicuous gaps. Airfall glass compositions also cluster into modes, and the presence of multiple modes indicates tapping of different magma volumes during early phases of eruption. Equilibrium assemblages of pigeonite and augite are used to reconstruct compositional and thermal gradients in the pre-eruptive reservoir. The recurrence of identical compositional modes and of mineral pairs equilibrated at high temperatures in successive eruptive units is consistent with the persistence of their respective liquids in the magma reservoir. Recurrence intervals of identical modes range from 0.3 to 0.9 Myr and suggest possible magma residence times of similar duration. Eruption ages, magma temperatures, Nd isotopes, and pyroxene and glass compositions are consistent with a long-lived, dynamically evolving magma reservoir that was chemically and thermally zoned and composed of multiple discrete magma volumes.

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The Bruneau-Jarbidge eruptive center (BJEC) in the central Snake River Plain, Idaho, USA consists of the Cougar Point Tuff (CPT), a series of ten, high-temperature (900-1000°C) voluminous ignimbrites produced over the explosive phase of volcanism (12.8-10.5 Ma) and more than a dozen equally high-temperature rhyolite lava flows produced during the effusive phase (10.5-8 Ma). Spot analyses by ion microprobe of oxygen isotope ratios in 210 zircons demonstrate that all of the eruptive units of the BJEC are characterized by zircon δ¹⁸O values ≤ 2.5‰, thus documenting the largest low δ¹⁸O silicic volcanic province known on Earth (>10⁴ km³). There is no evidence for voluminous normal δ¹⁸O magmatism at the BJEC that precedes generation of low δ¹⁸O magmas as there is at other volcanic centers that generate low δ¹⁸O magmas such as Heise and Yellowstone. At these younger volcanic centers of the hotspot track, such low δ¹⁸O magmas represent ~45 % and ~20% respectively of total eruptive volumes. Zircons in all BJEC tuffs and lavas studied (23 units) document strong δ¹⁸O depletion (median CPT δ¹⁸OZrc = 1.0‰, post-CPT lavas = 1.5‰) with the third member of the CPT recording an excursion to minimum δ¹⁸O values (δ¹⁸OZrc= -1.8‰) in a supereruption > 2‰ lower than other voluminous low δ¹⁸O rhyolites known worldwide (δ¹⁸OWR ≤0.9 vs. 3.4‰). Subsequent units of the CPT and lavas record a progressive recovery in δ¹⁸OZrc to ~2.5‰ over a ~ 4 m.y. interval (12 to 8 Ma). We present detailed evidence of unit-to-unit systematic patterns in O isotopic zoning in zircons (i.e. direction and magnitude of Δcore-rim), spectrum of δ¹⁸O in individual units, and zircon inheritance patterns established by re-analysis of spots for U-Th-Pb isotopes by LA-ICPMS and SHRIMP. In conjunction with mineral thermometry and magma compositions, these patterns are difficult to reconcile with the well-established model for "cannibalistic" low δ¹⁸O magma genesis at Heise and Yellowstone. We present an alternative model for the central Snake River Plain using the modeling results of Leeman et al. (2008) for ¹⁸O depletion as a function of depth in a mid-upper crustal protolith that was hydrothermally altered by infiltrating meteoric waters prior to the onset of silicic magmatism. The model proposes that BJEC silicic magmas were generated in response to the propagation of a melting front, driven by the incremental growth of a vast underlying mafic sill complex, over a ~5 m.y. interval through a crustal volume in which a vertically asymmetric δ¹⁸OWR gradient had previously developed that was sharply inflected from ~ -1 to 10‰ at mid-upper crustal depths. Within the context of the model, data from BJEC zircons are consistent with incremental melting and mixing events in roof zones of magma reservoirs that accompany surfaceward advance of the coupled mafic-silicic magmatic system.

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A general method for the preparation of novel disulfide-tethered macrocyclic diacylglycerols (DAGs) has been described. Overall synthesis involved stepwise protection, acylation, and deprotection to yield the bis(omega-bromoacyl) glycerols. In the crucial macrocyclization step, a unique reagent, benzyltriethylammonium tetrathiomolybdate (BTAT), has been used to convert individual bis(omega-bromoacyl) glycerols to their respective macrocyclic disulfides. DAG 6, which had ether linkages between hydrocarbon chains and the glycerol backbone, was also synthesized from an appropriate precursor using a similar protocol. One of the DAGs (DAG 5) had a carbon-carbon tether instead of a disulfide one and was synthesized using modified Glaser coupling. Preparation of alpha-disulfide-tethered DAG (DAG 4) required an alternative method, as treatment of the bisbromo precursor with BTAT gave a mixture of several compounds from which separation of the target molecule was cumbersome. To avoid this problem, the bisbromide was converted to its corresponding dithiocyanate, which on further treatment with BTAT yielded the desired DAG (DAG 4) in good yield. Upon treatment with the reducing agent dithiothreitol (DTT), the DAGs that contain a disulfide tether could be quantitatively converted to their "open-chain" thiol analogues. These macrocyclic DAGs and their reduced "open-chain" analogues have been incorporated in DPPC vesicles to study their effect on model membranes. Upon incorporation of DAG 1 in DPPC vesicles, formation of new isotropic phases was observed by P-31 NMR, These isotropic phases disappeared completely on opening the macrocyclic ring by a reducing agent. The thermotropic properties of DPPC bilayers having DAGs (1-6) incorporated at various concentrations were studied by differential scanning calorimetry. Incorporation of DAGs in general reduced the cooperativity unit (CU) of the vesicles. Similar experiments with reduced "open-chain" DAGs incorporated in a DPPC bilayer indicated a recovery of CU with respect to their macrocyclic "disulfide" counterparts. The effect of inclusion of these DAGs on the activity of phospholipase A(2) (PLA(2)) was studied in vitro. Incorporation of DAC 1 in DPPC membranes potentiated both bee venom and cobra venom PLA(2) activities.

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The impacts of 4 grazing strategies (year-long grazing, summer grazing, winter grazing and winter grazing plus spring burning) on the grass:legume balance were studied between 2000 and 2006 in a pasture oversown with Stylosanthes scabra cv. Seca (Seca stylo) in central Queensland. Seasonal rainfall throughout the study was generally below average. Total pasture yields in autumn were higher in the 2 winter grazing than the 2 summer grazing treatments, largely reflecting the sampling time relative to when grazing occurred. There were few differences in Seca composition in autumn, although there was a clear trend for Seca composition to be reduced by winter grazing plus burning. Both the frequency of occurrence and plant density of Seca were higher under the 2 summer grazing treatments and there was also a trend for the density of juvenile plants (<5 cm height) to be higher in the 2 summer grazing treatments. Seca soil seed banks were generally low and were reduced in the winter grazing plus burning treatment in spring 2002. The frequency of the palatable perennial grass Pennisetum ciliaris (Biloela buffel grass) was reduced while that of the 'increaser' species Bothriochloa pertusa (Indian couch grass) and Stachytarpheta jamaicensis (snake weed) increased in the 2 summer grazing treatments compared with the 2 winter grazing treatments. Burning in spring increased soil loss in treatments grazed in winter. Differences in Seca frequency and density but not composition were explained by the 2 summer grazing treatments promoting 'gaps' in the pasture which were then colonised by Seca plants and other 'increaser' species. It was reasoned that, with time, mature Seca plants in the 2 winter grazing treatments would die so that Seca composition would eventually become higher under summer grazing regimes than under winter grazing. It was concluded that limiting grazing to particular seasons can alter legume:grass balance and that a time-frame of 5-8 years with average to good rainfall would be necessary to achieve large shifts in composition.

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An assessment of marine elapid snakes found 9% of marine elapids are threatened with extinction, and an additional 6% are Near Threatened. A large portion (34%) is Data Deficient. An analysis of distributions revealed the greatest species diversity is found in Southeast Asia and northern Australia. Three of the seven threatened species occur at Ashmore and Hibernia Reefs in the Timor Sea, while the remaining threatened taxa occur in the Philippines, Niue, and Solomon Islands. The majority of Data Deficient species are found in Southeast Asia. Threats to marine snakes include loss of coral reefs and coastal habitat, incidental bycatch in fisheries, as well as fisheries that target snakes for leather. The presence of two Critically Endangered and one Endangered species in the Timor Sea suggests the area is of particular conservation concern. More rigorous, long-term monitoring of populations is needed to evaluate the success of "conservation measures" for marine snake species, provide scientifically based guidance for determining harvest quotas, and to assess the populations of many Data Deficient species.

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Spider venoms contain a plethora of insecticidal peptides that act on neuronal ion channels and receptors. Because of their high specificity, potency and stability, these peptides have attracted much attention as potential environmentally friendly insecticides. Although many insecticidal spider venom peptides have been isolated, the molecular target, mode of action and structure of only a small minority have been explored. Sf1a, a 46-residue peptide isolated from the venom of the tube-web spider Segesteria florentina, is insecticidal to a wide range of insects, but nontoxic to vertebrates. In order to investigate its structure and mode of action, we developed an efficient bacterial expression system for the production of Sf1a. We determined a high-resolution solution structure of Sf1a using multidimensional 3D/4D NMR spectroscopy. This revealed that Sf1a is a knottin peptide with an unusually large β-hairpin loop that accounts for a third of the peptide length. This loop is delimited by a fourth disulfide bond that is not commonly found in knottin peptides. We showed, through mutagenesis, that this large loop is functionally critical for insecticidal activity. Sf1a was further shown to be a selective inhibitor of insect voltage-gated sodium channels, consistent with its 'depressant' paralytic phenotype in insects. However, in contrast to the majority of spider-derived sodium channel toxins that function as gating modifiers via interaction with one or more of the voltage-sensor domains, Sf1a appears to act as a pore blocker.