Indicator-free electrochemical DNA hybridization biosensor

A new electrochemical hybridization biosensor protocol without an external indicator is described. The biosensor format involves the immobilization of inosine-substituted (guanine-free) probe onto the carbon paste transducer, and a direct chronopotentiometric detection of the duplex formation by the appearance of the guanine oxidation peak of the target. Such a use of the intrinsic DNA electrochemical response for monitoring hybridization events offers several advantages (over the common use of external indicators), including the appearance of a new peak, a Aat background, or simplicity. A 4 min short hybridization period allows a detection limit around 120 ng/ml. Performance characteristics of the sensor are described along with future prospects. (C) 1998 Elsevier B.V. B.V. All rights reserved.

Thermal analysis as a screening technique for the characterization of babassu flour and its solid fractions after acid and enzymatic hydrolysis

The babassu (Orbignya phalerata) is a native tree found in northern Brazil. Extracts of the babassu coconut have been widely used in industry. Babassu flour has about 60% starch, thus, besides nourishment it can be used as an alternative biofuel source. However, the properties of this starch lack of study and understanding. The main purpose of this study was to investigate the thermal behavior of raw babassu flour and its solid hydrolyzed fraction. The analyses were carried out using SHIMADZU DSC and TG thermic analyzers. The results demonstrated a reduction in thermal stability of the solid hydrolyzed fraction compared to raw matter. The kinetic parameters were investigated using non-isothermal methods and the parameters obtained for its decomposition process were an E(a) of 166.86 kJ mol(-1) and a frequency factor (beta) of 6.283 x 1014 min(-1); this was determined to be a first order reaction (n = 1). (C) 2011 Elsevier B.V. All rights reserved.

Effects of vitamin E supplementation on renal non-enzymatic antioxidants in young rats submitted to exhaustive exercise stress

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

Freshwater stingrays: study of epidemiologic, clinic and therapeutic aspects based on 84 envenomings in humans and some enzymatic activities of the venom

Freshwater stingrays are very common in the Parana, Paraguay, Araguaia, and Tocantins Rivers and tributaries in Brazil. This study presents the clinical aspects of 84 patients injured by freshwater stingrays. Intense pain was the most conspicuous symptom. Skin necrosis was observed in a high percentage of the victims, mostly fishermen and bathers. The initial therapeutic procedures, like immersion of the affected member in hot water were effective in the initial phases of the envenoming, especially in the control of the acute pain; however, they did not prevent skin necrosis. By SDS-PAGE, the freshwater stingray (Potamotrygon falkneri) venom extract presented a major band of approximately 12 kDa. Several other components distributed between 15 and 130 kDa were detected in the venom extract. Many components with molecular mass above 80 and 100 kDa have gelatinolytic and caseinolytic activities, respectively. Hyaluronidase activity was detected only in a component around 84 kDa in P. falkneri venom extract. Our results demonstrated that the presence of these enzymes could explain partially the local clinical pictures presented by patients wounded by freshwater stingray. (C) 2004 Elsevier Ltd. All rights reserved.

Morphological and enzymatic analysis of the midgut of Anopheles darlingi during blood digestion

The midgut of adult female Anopheles darlingi is comprised of narrow anterior and dilated posterior regions, with a single layered epithelium composed by cuboidal digestive cells. Densely packed apical microvilli and an intricate basal labyrinth characterize each cell pole. Before blood feeding, apical cytoplasm contains numerous round granules and whorled profiles of rough endoplasmic reticulum. Engorgement causes a great distension of midgut. This provokes the flattening of digestive cells and their nuclei. Simultaneously, apical granules disappear, the whorls of endoplasmic reticulum disassemble and 3 h post bloodmeal (PBM), nucleoli enlarge manyfold. An intense absorptive process takes place during the first 24h PBM, with the formation of large glycogen inclusions, which persist after the end of the digestive process. Endoproteases activities are induced after bloodmeal and attain their maximum values between 10 and 36 h PBM. At least two different aminopeptidases seem to participate in the digestive process, with their maximum activity values at 36 and 48 h PBM, respectively. Coarse electrondense aggregates, possibly debris from digested erythrocytes, begin to appear on the luminal face of the peritrophic membrane from 18 h PBM and persist during all the digestive process, and are excreted at its end. We suggest that these aggregates could contain some kind of insoluble form of haem, in order of neutralize its toxicity. (c) 2005 Elsevier Ltd. All rights reserved.

In situ evaluation of anticancer drug methotrexate-DNA interaction using a DNA-electrochemical biosensor and AFM characterization

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Enzymatic activity of hypopharyngeal gland extractsfrom workers of Apis mellifera (Hymenoptera, Apidae, Apinae)

This research presents a comparative study of enzymatic activity of the hypopharyngeal gland extracts from workers of Apis mellifera in three physiologic stages: newly emerged, nurse and forager workers, with the objective of contributing to the comprehension of the gland function. In order to determinate the enzymes present in the extracts, the Api Zym kit (Bio Merieux) was used to test the activity of 19 different enzymes. The enzymes found in larger amounts only in the hypopharyngeal glands from certain individuals were the following: in newly emerged workers, the N-acetyl-double down arrow-glucosaminidase that may be digesting the chitin of some food ingested by the bee; in forager workers, the acid phosphatase that is likely acting in authophagic processes, the a-glucosidase, in the processing of nectar into honey, and the double down arrow-glucosidases, in the pollen digestion.

Biochemical and cytochemical studies of the enzymatic activity of the venom glands of workers of honey bee Apis mellifera L. (Hymenoptera, Apidae)

Biochemical studies revealed that the activity of some hydrolytic enzymes from the venom glands of honey bee Apis mellifera was higher in workers of 14 days of age than in those of 40 days. Among these enzymes, the highest activity was recorded for acid phosphatase, which was cytochemically detected throughout the length of the secretory filament and surrounding the canaliculi of the distal region of the reservoir. The acid phosphatase was considered to be a typical secretion product, since it was present in the cytoplasm as well as in the canaliculi of the secretory cells. (c) 2009 Elsevier Ltd. All rights reserved.

Production and characterization of a milk-clotting protease in the crude enzymatic extract from the newly isolated Thermomucor indicae-seudaticae N31 (Milk-clotting protease from the newly isolated Thermomucor indicae-seudaticae N31)

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

Evaluation of the beta-glucanolytic enzyme complex of Trichoderma harzianum Rifai for the production of gluco-oligosaccharide fragments by enzymatic hydrolysis of 1,3;1,6-beta-D-glucans

Botryosphaeran, a new exopolysaccharide from the endophytic fungus Botryosphaeria rhodina MAMB-05, and algal laminarin were hydrolyzed by partially-fractionated enzymes of the beta-glucanolytic complex from Trichoderma harzianum Rifai. beta-Glucanase fractions (F-I and F-II) separated by gel permeation chromatography presented different modes of attack on botryosphaeran and laminarin. Botryosphaeran was hydrolyzed to the extent of 66% (F-I) and 98% (F-II) within 30 min, and its main hydrolysis products were gluco-oligosaccharides of DP >= 4, with lesser amounts of glucose, di- and tri-saccharides. The action of enzyme fractions I and II on laminarin resulted in 15% conversion to glucose, while the percentage of saccharification was radically different (70% for F-I and 25% for F-II). The different product arrays within the polysaccharide hydrolysates can be explained by the difference in the enzymes' specificities within each enzyme fraction, and the molecular structures of the polysaccharides and their complexity.

Effect of enzymatic hydrolysis on some physicochemical properties of root and tuber granular starches

A hidrólise enzimática do amido pode fornecer informações importantes sobre sua estrutura granular. Amidos de mandioca, batata-doce, mandioquinha-salsa e batata foram hidrolisados por α-amilase bacteriana a 37 °C durante 48 horas, e algumas propriedades físico-químicas dos resíduos da hidrólise foram determinadas. O amido de mandioca foi o mais suscetível à enzima com 20,9% de hidrólise, enquanto o amido de batata foi o mais resistente com 5,9%. O tamanho médio dos grânulos variou de 10,8 a 23,4 μm para os amidos de mandioquinha-salsa e batata, respectivamente. Amidos de mandioca e batata-doce apresentaram um padrão de difração de raio-X tipo A, enquanto os amidos de mandioquinha-salsa e batata mostraram padrão tipo B. Todos os amidos nativos mostraram superfície granular lisa e, após hidrólise, os amidos de mandioca, batata-doce e mandioquinha-salsa mostraram alguns grânulos bastante degradados, enquanto o amido de batata apresentou sutil sinal de degradação. O teor de amilose dos amidos diminuiu com a hidrólise para os amidos de mandioca, batata-doce e mandioquinha-salsa, permanecendo inalterado para o amido de batata. Como esperado, a viscosidade intrínseca e as propriedades de pasta diminuíram para todos os amidos hidrolisados. Não houve diferença significativa entre as propriedades térmicas dos amidos nativos e hidrolisados. Estes resultados sugeriram que a hidrólise ocorreu nas áreas cristalinas e amorfas dos grânulos. O padrão de difração do tipo B e o grande tamanho dos grânulos do amido de batata podem ter contribuído para a maior resistência deste amido à hidrólise.

Enzymatic toxins from snake venom: structural characterization and mechanism of catalysis

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

Enzymatic Activity, Sensitivity to Antifungal Drugs and Baccharis dracunculifolia Essential Oil by Candida Strains Isolated from the Oral Cavities of Breastfeeding Infants and in Their Mothers' Mouths and Nipples

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