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Taxonomia e diversidade dos Alysiinae (Hymenoptera: Braconidae) Neotropicais, com ênfase na fauna da Mata Atlântica, Brasil
(Universidade Federal de São Carlos, 2005-02-17) Arouca, Raquel Gonçalves; Dias, Angélica Maria Penteado Martins; https://lattes.cnpq.br/8637344058859147
The material studied comes from eighteen localities of Atlantic Forest from BIOTA/FAPESP Project: João Pessoa, PB; Recife, PE; Quebrangulo, AL; Santa Luzia do Itanhy, SE; Mata São João, BA; Mata da Esperança, BA: Porto Seguro, BA: Linhares, ES; Santa Teresa, ES; Santa Maria Madalena, RJ; Nova Iguaçu, RJ; Ubatuba, SP; Salesópolis, SP; Ribeirão Grande, SP; Peruíbe, SP; Morretes, PR; São Francisco do Sul, SC and São Bento do Sul, SC. Other specimens from the DCBU collection (Departamento de Ecologia e Biologia Evolutiva, Universidade Federal de São Carlos) were studies too. The material was collected using three techniques: Malaise traps, Moericke traps and sweeping vegetation. The amount of Alysiinae examined was 2.920 specimens (2.082 of BIOTA/FAPESP Project and 838 of other collection), identified in 14 genera. Dinotrema, Aphaereta e Phaenocarpa were the most frequent genera. Indices of Diversity and Equitability were used to discuss the richness and dominance of genera in each locality. The higher values of richness were recorded in Nova Iguaçu, RJ; São Bento do Sul, SC; São Francisco do Sul, SC; Santa Teresa, ES e Santa Luzia do Itanhy, SE and the lower in João Pessoa, PB; Recife, PE; Mata São João, BA; Porto Seguro, BA e Linhares, ES. These data evidenced a trend to the enrichment of the Alysiinae fauna to the Atlantic Forest areas in largest latitudes. The higher Diversity Index was recorded in Santa Luzia do Itanhy, SE and the lower in João Pessoa, PB. Linhares, ES presented the highest Equitability and Santa Teresa, ES the lower. Cluster Analysis was adopted taking as attribute the occurrence of the Alysiiinae genera. Four species of Phaenocarpa and Aphaereta confusa Wharton, 1994 female were described. Eight species were reported from Brazil for the first time: Gnathopleura carinata (Szépligeti, 1904); G. bugabensis (Cameron, 1887); G. semirufa (Brullé, 1846); Ilatha pulchripennis (Latreille, 1805); Idiasta delicata (Papp, 1969); Phaenocarpa pericarpa Wharton & Carrejo, 1999; P. cratomorpha Wharton, 1994 and Aphaereta confusa Wharton, 1994; and four new genera were identified. Those results show the importance and the need of the taxonomic and ecological studies on the subfamily Alysiinae, especially in the Neotropical region.
Desenvolvimento de procedimentos analíticos para a determinação de N-acetilcisteína em produtos farmacêuticos
(Universidade Federal de São Carlos, 2005-02-17) Suarez, Willian Toito; Fatibello Filho, Orlando; https://lattes.cnpq.br/9859737944357808; https://lattes.cnpq.br/6680603267238159
In this dissertation four analytical procedures for the determination of Nacetylcysteine in pharmaceutical formulations are described. The first procedure developed was a flow injection analysis system for turbidimetric determination of Nacetylcysteine employing Ag+ ions in an acid medium as the precipitant reagent. In this system, 250 µL of 0.01 mol L-1 AgNO3 solution and 500 µL of sample solution were inserted simultaneously into a merging zones flow system. After the precipitate formation in a 100 cm coil reactor, the precipitate generated was monitored turbidimetrically at 400 nm. Desionised water flowing intermittently at 6.3 mL min-1 was used to wash out the precipitate during the sampling stage. The analytical curve was linear in the N-acetylcysteine concentration range from 1.0 x 10-4 to 1.0 x 10-3 mol L-1; with a detection limit of 8.0 x 10-5 mol L-1 (3σB/slope) and sampling frequency of 60 h-1 was obtained. The relative standard deviation was smaller than 1% for Nacetylcysteine solutions in the concentrations of 1.0 x 10-4 and 5.0 x 10-4 mol L-1 (n=20). The recoveries obtained for two samples ranged from 104 to 122%. A flow injection system with spectrophotometric detection is proposed for determining Nacetylcysteine in pharmaceutical formulations. In this system, N-acetylcysteine was oxidized by Fe(III) and the Fe(II) produced is spectrophotometrically monitored as Fe(II)-1,10-phenantroline complex at 515 nm. Under the optimum analytical conditions, the linearity of the calibration graph for N-acetylcysteine ranged from 1.8 x 10-5 to 1.5 x 10-4 mol L-1. The detection limit of 6.3 x 10-6 mol L-1 (3σB/slope) and recoveries between 102 to 113 % were obtained. The preparation and electrochemical characterization of a carbon paste electrode modified with copper (II) hexacyanoferrate(III) (CuCHF) as well as its behaviour as electrocatalyst toward the oxidation of N-acetylcysteine were investigated. The electrochemical behaviour of the modified electrode and the electrooxidation of N-acetylcysteine were explored using sweep linear voltammetry. The best voltammetric response was observed for a paste composition of 20%(w/w) copper (II) hexacyanoferrate(III) complex, acetate buffer solution at pH of 6.0 as the electrolyte and scan rate of 10 mV s-1. A linear voltammetric response for N-acetylcysteine was obtained in the concentration range xiv from 1.2 x 10-4 to 8.3 x 10-4 mol L-1, with a detection limit of 6.3 x 10-5 mol L-1 (3σB/slope). The proposed electrode is useful for the quality control and routine analysis of N-acetylcysteine in pharmaceutical formulations. Finally, a simple, precise, rapid and low-cast potentiometric method for N-acetylcysteine determination in pure form and in pharmaceutical preparations is proposed. N-acetylcysteine present in tablets containing known quantity of drug was potentiometrically titrated in aqueous solution with AgNO3. No interferences were observed in the presence of common components of the tablets as saccharin, sucrose and EDTA. The analytical results obtained by applying the proposed method compared very favorably with those obtained by the comparative method. Recovery of N-acetylcysteine from various tablets dosage formulations range from 98.7 to 103.0%. Compared to others procedures reported in the literature the procedures developed in this dissertation shows to be better and cheaper to determination of N-acetylcysteine in pharmaceutical formulations.
Obtenção de uma desintegrina recombinante de Agkistrodon contortrix laticinctus e estudo dos efeitos de desintegrinas na expressão do fator de crescimento de endotélio vascular
(Universidade Federal de São Carlos, 2005-02-17) Ribeiro, Juliana Uema; Araújo, Heloísa Sobreiro Selistre de; https://lattes.cnpq.br/4065824911933203; https://lattes.cnpq.br/4065824911933203
Disintegrins are snake venom proteins used in the study of integrin-mediated cellular adhesion through integrins. Recombinant disintegrin production may notable supply faster, larger and most homogeneous protein samples than bruit venom processing. This work describes optimization of the recombinant ACLD-C expression technique. ACLD-C is a disintegrin-like protein from Agkistrodon contortrix laticinctus, that have a disintegrin-like domain with DCD motif, and a cysteine-rich domain. This protein was subcloned from the ACLD clone (GenBank U86634), a metallopeptidase. A protocol of expression was established in our laboratory, where recombinant ACLD-C is produced in E. coli, using the expression vector pMal-p. This allows protein target expression in form of N-terminal fusion with MBP (maltose-binding protein), that facilitate purification, correct folding and production of disintegrin in soluble form. In fact, MBP/ACLD-C was was expressed in soluble form and purified in two stages: affinity comatography in amilose resin and cromatography of gel-filtration in superdex-200 resin. Modifications in expression protocol optimized MBP/ACLD-C expression in culture of 0,4 mg/L of culture of 1,2 mg/L (12,6% total of protein in periplasmic space of E.coli). MBP/ACLD-C imobilizated in amilose resin was subjected to digestion with enzyme factor Xa to separate ACLD-C of MBP. The separation was confirmed through positive reaction with anti-ALT-C antibody. ALT-C is a disintegrin-like protein from Bothrops alternatus. ALT-C induced both expression of VEGF in human fibroblasts and proliferation of endothelial cells and angiogenesis. In second part of this work, we investigated if MBP/ACLD-C was also able to induce the expression of this growth factor in this cells. MBP/ACLD-C and two other disintegrins, echistatina, a disintegrin-RGD, and EC6, a heterodimeric disintegrin, were incubated with fibroblasts. The influency of these proteins in VEGF expression was evaluated by ELISA. MBP/ACLD-C was able to strongly induce the expression of VEGF until 4 hours, when the level decreased until 24 hours. Experiment in period of 24 48 hours was not done due to problems of contaminations. Echistatin did not induced VEGF expression, but induced fibroblasts dettachment from plastic. The latter event was not observed in MBP/ACLD-C or EC6 incubated cells.