Comparative RH Maps of the River Buffalo and Bovine Y Chromosomes

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

Expression and processing of recombinant sarafotoxins precursor in Pichia pastoris

Sarafotoxins are peptides isolated from the Atractaspisw snake venom. with strong constrictor effect on cardiac and smooth muscle. They are structurally and functionally related to endothelins. The sarafotoxins precursor cDNA predicts an unusual structure 'rosary-type', with 12 successive similar stretches of sarafotoxin (SRTX) and spacer, in the present work, the recombinant precursor of SRTXs was sub-cloned and expressed in the yeast Pichia pastoris. and secreted to the culture medium, Characterization by SDS-PAGE, immunoblot, mass spectrometry and biological activity, suggests that intact precursor was expressed but processing into mature toxins also occurred. Furthermore, our results indicate that the correct proportion of sarafotoxin types as contained in the precursor, is obtained in the yeast culture medium. Contractile effects of the expressed toxins, on rat and Bothrops jararaca isolated aorta, were equivalent to 5 X 10(-10) M and 5 x 10(-11) M of sarafotoxin b, respectively. The enzymes responsible for the complete maturation of sarafotoxins precursor are still unknown. Our results strongly suggest that the yeast Pichia pastoris is able to perform such a maturation process. Thus, the yeast Pichia pastoris may offer an alternative to snake venom gland to tentatively identify the molecular process responsible for SRTXs release. (C) 2001 Elsevier B.V. Ltd. All rights reserved.

Expression and purification of human respiratory syncytial virus recombinant fusion protein

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

Preliminary radiation hybrid map for river buffalo chromosome 6 and comparison to bovine chromosome 3

We present the first radiation hybrid (RH) map of river buffalo (Bubalus bubalis) chromosome 6 (BBU6) developed with a recently constructed river buffalo whole-genome RH panel (BBURH5000). The preliminary map contains 33 cattle-derived markers, including 12 microsatellites, 19 coding genes and two ESTs, distributed across two linkage groups. Retention frequencies for markers ranged from 14.4% to 40.0%. Most of the marker orders within the linkage groups on BBU6 were consistent with the cattle genome sequence and RH maps. This preliminary RH map is the starting point for comparing gene order between river buffalo and cattle, presenting an opportunity for the examination of micro-rearrangements of these chromosomes. Also, resources for positional candidate cloning in river buffalo are enhanced.

Sensory acceptability of raw and extruded bovine rumen protein in processed meat products

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

Correlation of temperature induced conformation change with optimum catalytic activity in the recombinant G/11 xylanase A from Bacillus subtilis strain 168 (1A1)

The 1.7 angstrom resolution crystal structure of recombinant family G/11 beta-1,4-xylanase (rXynA) from Bacillus subtilis 1A1 shows a jellyroll fold in which two curved P-sheets form the active-site and substrate-binding cleft. The onset of thermal denaturation of rXynA occurs at 328 K, in excellent agreement with the optimum catalytic temperature. Molecular dynamics simulations at temperatures of 298-328 K demonstrate that below the optimum temperature the thumb loop and palm domain adopt a closed conformation. However, at 328 K these two domains separate facilitating substrate access to the active-site pocket, thereby accounting for the optimum catalytic temperature of the rXynA. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

The Water Effect on the Kinetics of the Bovine Liver Catalase

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Partial characterization of protease from a thermophilic fungus, Thermoascus aurantiacus, and its hydrolytic activity on bovine casein

Proteases are one of the most important groups of industrial enzymes, with considerable application in the food industry. The aim of this work was to study a novel protease produced by the thermophilic fungus, Thermoascus aurantiacus, through solid-state fermentation (SSF). The enzyme acted optimally at pH 5.5 and 60 degrees C it was stable up to 60 degrees C for 1 h and in the pH range 3.0-9.5. To elucidate the enzyme's proteolytic activity, its hydrolytic profile on bovine casein, an important protein in the food industry, was studied by enzymatic hydrolysis on skim milk, analyzed by gel electrophoresis (UREA-PAGE), which clearly showed that the protease does not have the same specificity as bovine chymosin. (c) 2006 Elsevier Ltd. All rights reserved.

Influence of the association between simvastatin and demineralized bovine bone matrix on bone repair in rats

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

Evaluation of the shear bond strength of a composite resin in sound, demineralized and hypermineralized bovine dentin after irradiation with Er:YAG and Nd:YAG lasers, employing a self-etching adhesive system

Background and Objectives. The adhesion of dental materials is important for the success of treatment. The aim of this study is to evaluate the bond strength of a composite resin applied with a self-etching adhesive system in different dentins after irradiation with Er:YAG and Nd:YAG lasers, observing their morphologic pattern using Scanning Electronic Microscopy (SEM). Materials and Methods. The buccal surface of 72 bovine incisors was worn until exposure of medium depth dentin. The specimens were divided into three groups; GI: normal, GII: demineralized and GIII: hypermineralized dentin. These were also divided into two subgroups; A-irradiated for 30 s with Er:YAG laser in noncontact mode at 40 mJ and 6 Hz and B- irradiated for 30 s with Nd:YAG laser in contact mode at 60 mJ and 10 Hz. The adhesive system Clearfil SE. Bond (Kuraray) and composite resin Tetric Ceram (Vivadent) were applied on the irradiated area by the incremental technique. After storage for 24 h in distilled water at 37 degrees C, the specimens were submitted to the shear strength test in a universal testing machine (EMIC) at a crosshead speed of 1.0 mm/min. Other specimens were made to be analyzed by SEM. Results. The results were statistically analyzed by Analysis of Variance and the Tukey test. Regardless of the type of dentin, the bond strength of specimens irradiated with the Nd:YAG laser (8,94 +/- 2,07) was higher compared to specimens irradiated with the Er:YAG laser (7,03 +/- 2,47); the highest bond strength was obtained for the group of hypermineralized dentin irradiated with the Nd:YAG laser. The SEM analysis showed that the Er:YAG laser caused opening of tubules and the Nd:YAG laser produced areas of fusion as well as regions of opening of dentinal tubules. Conclusions. The dentin showed different morphological patterns and the laser promote alterations on their surfaces, influencing the bond strength of the composite resin. (C) 2010 Laser Institute of America.