Purification, characterization, crystallization and theoretical molecular modeling of gyroxin fraction from Crotalus durissus terrificus venom (Laurenti, 1768)

The venom of Crotalus durissus terrificus snakes presents various substances, including a serine protease with thrombin-like activity, called gyroxin, that clots plasmatic fibrinogen and promote the fibrin formation. The aim of this study was to purify and structurally characterize the gyroxin enzyme from Crotalus durissus terrificus venom. For isolation and purification, the following methods were employed: gel filtration on Sephadex G75 column and affinity chromatography on benzamidine Sepharose 6B; 12% SDS-PAGE under reducing conditions; N-terminal sequence analysis; cDNA cloning and expression through RT-PCR and crystallization tests. Theoretical molecular modeling was performed using bioinformatics tools based on comparative analysis of other serine proteases deposited in the NCBI (National Center for Biotechnology Information) database. Protein N-terminal sequencing produced a single chain with a molecular mass of similar to 30 kDa while its full-length cDNA had 714 bp which encoded a mature protein containing 238 amino acids. Crystals were obtained from the solutions 2 and 5 of the Crystal Screen Kit (R), two and one respectively, that reveal the protein constitution of the sample. For multiple sequence alignments of gyroxin-like B2.1 with six other serine proteases obtained from snake venoms (SVSPs), the preservation of cysteine residues and their main structural elements (alpha-helices, beta-barrel and loops) was indicated. The localization of the catalytic triad in His57, Asp102 and Ser198 as well as S1 and S2 specific activity sites in Thr193 and Gli215 amino acids was pointed. The area of recognition and cleavage of fibrinogen in SVSPs for modeling gyroxin B2.1 sequence was located at Arg60, Arg72, Gln75, Arg81, Arg82, Lis85, Glu86 and Lis87 residues. Theoretical modeling of gyroxin fraction generated a classical structure consisting of two alpha-helices, two beta-barrel structures, five disulfide bridges and loops in positions 37, 60, 70, 99, 148, 174 and 218. These results provided information about the functional structure of gyroxin allowing its application in the design of new drugs.

Components of Turnera diffusa Willd. var. afrodisiaca (Ward) Urb. essential oil

Turnera diffusa Willd. var. afrodisiaca (Ward) Urb. (syn. T aphrodisiaca) (Turneraceae) is a common aromatic plant growing wild in the subtropical regions of America and Africa. Its essential oil was studied by GC and GC-MS. Fifty-four components were characterized and identified, the most abundant being 1,8-cineol (11.4%), opoplenone (10.3%) cadalene (5.1%) and epi-cubenol (4.1%). Copyright (C) 2002 John Wiley Sons, Ltd.

Hematological changes in sheep inoculated with natural and Cobalt60-irradiated Crotalus durissus terrificus venom (Laurenti, 1768)

Natural (NV) and Cobalto60-irradiated (IrV) Crotalus durissus terrificus venom were used to evaluate serum production capacity of sheep and possible hematological and biochemical effects. Freeze-dried venom aliquots were diluted in acidified saline solution (NaCl 150 mM, pH 3.0) and irradiated by a Cobalt 60 source at a dose of 5.54 x 102 Gy/h and a concentration of 2.000 Gy. Twelve sheep were divided into two groups of six animals. One group received irradiated venom (IrV) and the other natural venom (NV). Three antigen doses (venom) were administered at monthly intervals. Blood samples were collected weekly for analysis of serum neutralization potency and capacity, complete blood count (CBC), total plasma protein, fibrinogen, albumin, and globulin. At the end of the experiment, the animals were challenged with a LD50 for sheep and showed no signs of envenoming. The two groups did not present clinical alterations. Results of the total leukocyte count did not present interaction or time factor effect for both groups, but there was a different action between them, with the NV group presenting more cells than the IrV group. The leukocyte increase to 13,000/ml indicates that slight leukocytosis occurred in the week after the first inoculation in the NV group. There was no statistically significant difference between groups in the absolute count of segmented neutrophils, eosinophils, and lymphocytes but there were statistically significant oscillations in values at the different collecting times. The NV group presented an increase in the absolute neutrophil count after the first inoculation that persisted for 5 weeks. In the IrV group, the increase in neutrophils occurred only in the first week returning to normal in the following weeks. The alterations in the neutrophil count are indicative of systemic inflammatory response related to cytokine release; response was more marked in the NV group, showing its greater toxicity.

Light-front zero-mode contribution to the Ward Identity

In a covariant gauge we implicitly assume that the Green's function propagates information from one point of the space-time to another, so that the Green's function is responsible for the dynamics of the relativistic particle. In the light front form one would naively expect, that this feature would be preserved. In this manner, the fermionic field propagator can be split into a propagating piece and a non-propagating (contact) term. Since the latter (contact) one does not propagate information; and therefore, supposedly can be discarded with no harm to the field dynamics we wanted to know what would be the impact of dropping it off. To do that, we investigated its role in the Ward identity in the light front. Here we use the terminology Ward identity to identify the limiting case of photon's zero momentum transfer in the vertex from the more general Ward-Takahashi identity with nonzero momentum transfer.

Liver dysfunction in patients bitten by Crotalus durissus terrificus (Laurenti, 1768) snakes in Botucatu (State of São Paulo, Brazil)

Os autores estudaram 32 doentes picados por serpentes venenosas, sendo 16 picados por Bothrops spp. e 16 por Crotalus durissus terrificus. Trinta doentes eram do sexo masculino e dois do feminino com idades variando entre 8 e 63 anos (méda 33±15). A prova da retenção da bromosulfaleína apresentou-se aumentada na maioria dos doentes picados por serpentes Crotalus durissus terrificus. Houve correlação positiva entre a retenção da bromosulfaleína e os níveis séricos de alanina aminotransferase e entre alanina e aspartato aminotransferase apenas nos doentes do grupo Crotalus. Um dos doentes evoluiu para o óbito e apresentou no exame anatomopatológico do fígado degeneração hidrópica e lesões mitocondriais. Os autores concluem que as alterações hepáticas são causadas por pelo menos dois mecanismos a saber: lesão mitocondrial por efeito do veneno crotálico; efeito das citoquinas, especialmente a interleucina-6.

Application of low-pressure gas chromatography-ion-trap mass spectrometry to the analysis of the essential oil of Turnera diffusa (Ward.) Urb.

Turnera diffusa Willd. var. afrodisiaca (Ward) Urb. (syn. T. aphrodisiaca) belongs to the family of Turneraceae and is an aromatic plant growing wild in the subtropical regions of America and Africa. It is widely used in the traditional medicine as e.g. anti-cough, diuretic, and aphrodisiac agent. This work presents a 3 min chromatographic analysis using low-pressure (LP) gas chromatography (GC)-ion-trap (IT) mass spectrometry (MS). The combination of a deactivated 0.6 m x 0.10 mm i.d., restrictor with a wide-bore CP-Wax 52 capillary column (10 m x 0.53 mm i.d., 1 mum) reduces the analysis time by a factor of 3-7 in comparison to the use of a conventional narrow bore column. Chromatographic conditions have been optimized to achieve the fastest separation with the highest signal/noise ratio in MS detection. These results allow fast and reliable quality control of the essential oil to be achieved. (C) 2003 Elsevier B.V. All rights reserved.

Purificação, caracterização, cristalização e modelagem molecular teórica da fração giroxina do veneno de Crotalus durissus terrificus (Laurenti 1768)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

As manifestações do gótico em Lovecraft: do Castelo de Otranto ao solar de Ward

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Heroísmos, sedições e heresias: a construção do ufanismo e do ressentimento nos sertões da capitania de São Paulo (1768-1774)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ecomorfologia de três espécies de Dipsas Laurenti, 1768 e Sibynomorphus Fitzinger, 1843 (Dipsadidae: Dipsadinae)

Fatores ecológicos como a utilização do ambiente e a alimentação, podem influenciar na morfologia corporal e cefálica de um organismo, porém, esta também pode ser o reflexo da linhagem filogenética a que ele pertence. A subfamília Dipsadinae inclui serpentes sul-americanas formadas por dois clados, um incluindo Geophis e Atractus e outro incluindo Dipsas, Ninia, Sibon e Sibynomorphus, que formam a tribo Dipsadini. As serpentes desta tribo apresentam um alto grau de modificações na morfologia corporal e cefálica relacionadas ao ambiente e a alimentação malacófaga. As espécies de Dipsas e Sibon possuem especializações relacionadas à utilização do ambiente arborícola, conferindo um melhor desempenho na locomoção pela vegetação, enquanto que as espécies de Sibynomorphus, em sua grande maioria, são adaptadas ao ambiente terrestre. Quanto à alimentação todas elas apresentam, em diferentes graus, uma série de modificações cranianas relacionadas à manipulação e ingestão de gastrópodes, lesmas e caramujos. Considerando que existe uma relação direta entre a forma da cabeça e a dieta, este trabalho tem como objetivo analisar as divergências da morfologia cefálica relacionadas à dieta entre espécies de Dipsas e Sibynomorphus, assim como analisar possíveis convergências entre espécies destes gêneros e Tomodon dorsatus, espécie da tribo Tachymenini, reconhecidamente especialista em lesmas. Para isto este trabalho está estruturado em duas partes: a primeira refere-se à Introdução geral, onde é apresentada uma ampla abordagem sobre as adaptações morfológicas relacionadas à alimentação dos Dipsadini, assim como as vantagens da utilização da técnica de morfometria geométrica em estudos morfológicos; e a segunda parte refere-se ao trabalho propriamente dito, intitulado “Ecomorfologia de três espécies de Dipsas Laurenti, 1768 e Sibynomorphus Fitzinger, 1843 (Dipsadidae: Dipsadinae)”.

Robert M. Ward Photograph Collection - Accession 502

The Robert M. Ward Photograph Collection consists of approximately 400 prints and 900 negatives from the photographs that he took in the 1940s, 1950s, and 1960s. Most of the events and individuals pictured are from the York County, SC area and include the Latta House in York, South Carolina, downtown Rock Hill, parades, baseball, military, aviation, sports, and other York County buildings and scenes. Also included are photographs of Landsford Canal State Park, York Technical College, Winthrop University, Catawba Indian chieftain King Haigler’s memorial, Andrew Jackson State Park, Kings Mountain State Park, Hall of Fame Golfer Sam Sneed's visit to Rock Hill, SC, and Charleston, South Carolina. The collection also contains newspaper articles from the York Observer, dated from 1988 to 1992, which helped to identify some of the people and events in the pictures.