249 resultados para haruki murakami
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Sodium (Na+) and chloride (Cl-) nutritional requirements, dietary electrolyte balance (DEB), and their effects on acid-base balance, litter moisture, and tibial dyschondroplasia (TD) incidence for young broiler chickens were evaluated in two trials. One-day-old Cobb broilers were distributed in a completely randomized design with six treatments, five replicates, and 50 birds per experimental unit. Treatments used in both experiments were a basal diet with 0.10% Na+ (Experiment 1) or Cl- (Experiment 2) supplemented to result in diets with Na+ or Cl- levels of 0.10, 0.15, 0.20, 0.25 ,0.30, or 0.35%, respectively. In Experiment 1, results indicated an optimum Na+ requirement of 0.26%. Sodium levels caused a linear increase in arterial blood gas parameters, indicating an alkalogenic effect of Na+. The hypertrophic area of growth plate in the proximal tibiotarsi decreased with Na+ levels. The TD incidence decreased with increases in dietary Na+. Litter moisture increased linearly with sodium levels. In Experiment 2, the Cl- requirement was estimated as 0.25%. Chloride levels caused a quadratic effect (P ≤ 0.01) on blood gas parameters, with an estimated equilibrium [blood base excess (BE) = 0] at 0.30% of dietary CT-. No Cl- treatment effects (P ≥ 0.05) were observed on litter moisture or TD incidence. The best DEB for maximum performance was 298 to 315 mEq/kg in Experiment 1 and 246 to 264 mEq/kg in Experiment 2. We concluded that the Na+ and Cl- requirements for optimum performance of young broiler chickens were 0.28 and 0.25%, respectively.
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A trial was carried to verify the effect of increasing levels (0, 10, 20, 30 and 40%) of canola meal in diets of broiler chickens on performance and mucous morphology, during the starting period (1 - 21 days). One thousand 1-d male chicks were assigned to a randomized experimental assay, with five treatments, four replicates and 50 birds per experimental unit. At the 1 st, 21 st and 41 st days, the birds and the rations were weighed. At the 21 st day, two birds of each experimental unit were sacrificed for collection of a duodenum segment to evaluate the intestinal morphometry. The results demonstrated that increasing levels of canola meal resulted in linear reduction on weight gain, weight and feed intake and worst feed conversion. During the growing period (21 to 41 days), when all birds received similar diet, average weight and weight gain decreased, as the canola meal levels increased, while feed conversion and feed intake were similar. Data regarding mucous morphology, submitted to the regression analysis, demonstrated that the crypt depth increased, as the canola meal levels increased. It is possible to include up to 20% of canola meal without damages on broiler performance and digestive tract.
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Lys49 snake-venom phospholipase A2 (PLA2) homologues are highly myotoxic proteins which, although lacking catalytic activity, possess the ability to disrupt biological membranes, inducing significant muscle-tissue loss and permanent disability in severely envenomed patients. Since the structural basis for their toxic activity is still only partially understood, the structure of myotoxin II, a monomeric Lys49 PLA2 homologue from Atropoides nummifer, has been determined at 2.08 Å resolution and the anion-binding site has been characterized. © 2006 International Union of Crystallography. All rights reserved.
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The results on the nutritional influence in the social hierarchy establishment among females from six colonies of Mischocyttarus cassununga show that: - there was no caste determination in the pre-imaginal period concerning ingested food quantities, however, in relation to qualification (proteins and carbohydrate) the Mann-Whitney test applied showed variation among colonies: in two colonics (colony 3, p=0.0209; colony 4, p=0.0090) the larvae which ingested a higher quantity of solid food became adults of hierarchically higher position, however in two other colonies, colony 1 (p=0.6744) and colony 2 (p=0.3367) this difference was not statistically significant; in two experimentally manipulated colonies the caste determination was observed in the imaginal phase (it was verified that the larger the food quantity consumed, the higher the assumed hierarchical position by the individuals); - adult individuals of hierarchically higher position in two colonies ingested a greater amount of solid than liquid food (Mann-Whitney; p ≤ 0.05), however in the other three colonies there was no difference. It can be seen that for all colonies (2, 3, 4, 5 and 6) the high tendency to solid food ingestion occurs. In colony 1, this data was not collected because the observation purpose was different); - inside the colony hierarchy, the older individuals were the ones that occupied higher hierarchical positions; - more than one inseminated female per colony was found in the two experimentally manipulated colonies ; - ovarian development is greater in inseminated females; - the average height of the 3rd tergite was greater for inseminated females, indicating an adaptation to accommodate larger ovary development (t-Student test; p ≤ 0.05).
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Some cytogenetical aspects of spermatozoa formation were studied in 9 Coreidae Brazilian species: Anasa bellator, Athaumastus haematicus, Chariesterus armatus, Dallacoris obscura, Dallacoris pictus, Leptoglossus gonagra, Leptoglossus zonatus, Sphictyrtus fasciatus, and Zicca annulata. Similarly to the other species described to date, all the species studied herein showed cystic spermatogenesis, a reddish membrane covering the testes, a X0 sex determining system, a pair of m-chromosomes, intersticial chiasmata in most autosomes, and autosomes dividing reductionally at first meiotic division and equationally in the second 1 while sex chromosomes, divide equationally and reductionally at first and second meiotic division, respectively. In addition, it was observed that the sex chromosome is heteropycnotic at prophase and that heteropycnotic chromosomal material is found in the nuclei at spermiogenesis. In the species studied, the diploid chromosome number ranged from 19 to 25. It was 19 in S. fasciatus (16A+2m+X0); 21 in A. bellator, A. haematicus, D. obscura, D. pictus, L. gonagra, and L. zonatus (18A+2m+X0); 23 in Z. annulata (20A+2m+X0); and 25 in C. armatus (22A+2m+X0). © 2007 The Japan Mendel Society.
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Includes bibliography
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Phospholipases D (PLDs) are principally responsible for the local and systemic effects of Loxosceles envenomation including dermonecrosis and hemolysis. Despite their clinical relevance in loxoscelism, to date, only the SMase I from Loxosceles laeta, a class I member, has been structurally characterized. The crystal structure of a class II member from Loxosceles intermedia venom has been determined at 1.7. Å resolution. Structural comparison to the class I member showed that the presence of an additional disulphide bridge which links the catalytic loop to the flexible loop significantly changes the volume and shape of the catalytic cleft. An examination of the crystal structures of PLD homologues in the presence of low molecular weight compounds at their active sites suggests the existence of a ligand-dependent rotamer conformation of the highly conserved residue Trp230 (equivalent to Trp192 in the glycerophosphodiester phosphodiesterase from Thermus thermophofilus, PDB code: 1VD6) indicating its role in substrate binding in both enzymes. Sequence and structural analyses suggest that the reduced sphingomyelinase activity observed in some class IIb PLDs is probably due to point mutations which lead to a different substrate preference. © 2011 Elsevier Inc.
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The genus Mischocyttarus comprises 245 species of neotropical basal eusocial wasps. They form small colonies (rarely more than few tens of individuals); castes are morphologically undifferentiated and determined behaviorally by agonistic interactions. The aim of this study was to verify the effects of the experimental disruption of social hierarchy on foraging activity of Mischocyttarus cerberus styx. We observed six colonies in postemergence phase and recorded data on the foraging activity under two experimental conditions: (1) removal of lower-ranked females and (2) removal of higher ranked females, except the queen. Our results showed that the removal of higher-ranked females had higher effect on the number of foraging trips of M. cerberus styx than the removal of lower-ranked females (the number of foraging trips/hour decreased by 66.4 and 32.7, resp.). Such results are likely due to the social organization of this species and the presence of a distinct class of females, which in this study were regarded as intermediates. Our data also showed that, irrespective of the hierarchical status of the females, the removal of two or three individuals affected significantly the number of foraging trips in this species. Copyright 2011 Vanderlei Conceio Costa Filho et al.
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This study aimed at developing radiographic techniques for the early detection of dyschondroplastic lesions in the tibia of broilers. The experiment was carried out at the facilities of UNIFOR/MG and Formiga and UNIFENAS/ Alfenas with 420 one-day-old male Cobb broilers. At 20 days of age, all birds were radiographed and identified with an alpha-numeric metal ring in the right leg. At 40 days of age, 42 broilers previously selected as a function of bone mineral density and lesion thickness scores were again radiographed and scored, and then sacrificed. Their right tibia was removed for gross and histological examination of the growth plate. The results showed that radiographic techniques are correlated with gross and histological examination and that there was no significant differences among techniques (P>0.05). it was concluded that the use of radiographic examination to identify tibial dyschondroplasia in broilers precludes the use of bone mineral density to diagnose this condition. The non-parametric statistical Chi-square test at 5% significance level was used to analyze the results.
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l-Amino acid oxidases (LAAOs) are flavoenzymes that catalytically deaminate l-amino acids to corresponding α-keto acids with the concomitant production of ammonia (NH 3) and hydrogen peroxide (H 2O 2). Particularly, snake venom LAAOs have been attracted much attention due to their diverse clinical and biological effects, interfering on human coagulation factors and being cytotoxic against some pathogenic bacteria and Leishmania ssp. In this work, a new LAAO from Bothrops jararacussu venom (BjsuLAAO) was purified, functionally characterized and its structure determined by X-ray crystallography at 3.1å resolution. BjsuLAAO showed high catalytic specificity for aromatic and aliphatic large side-chain amino acids. Comparative structural analysis with prokaryotic LAAOs, which exhibit low specificity, indicates the importance of the active-site volume in modulating enzyme selectivity. Surprisingly, the flavin adenine dinucleotide (FAD) cofactor was found in a different orientation canonically described for both prokaryotic and eukaryotic LAAOs. In this new conformational state, the adenosyl group is flipped towards the 62-71 loop, being stabilized by several hydrogen-bond interactions, which is equally stable to the classical binding mode. © 2012 Elsevier Inc.
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Catalytically inactive phospholipase A2 (PLA2) homologues play key roles in the pathogenesis induced by snake envenomation, causing extensive tissue damage via a mechanism still unknown. Although, the amino acid residues directly involved in catalysis are conserved, the substitution of Asp49 by Arg/Lys/Gln or Ser prevents the binding of the essential calcium ion and hence these proteins are incapable of hydrolyzing phospholipids. In this work, the crystal structure of a Lys49-PLA2 homologue from Bothrops brazili (MTX-II) was solved in two conformational states: (a) native, with Lys49 singly coordinated by the backbone oxygen atom of Val31 and (b) complexed with tetraethylene glycol (TTEG). Interestingly, the TTEG molecule was observed in two different coordination cages depending on the orientation of the nominal calcium-binding loop and of the residue Lys49. These structural observations indicate a direct role for the residue Lys49 in the functioning of a catalytically inactive PLA2 homologue suggesting a contribution of the active site-like region in the expression of pharmacological effects such as myotoxicity and edema formation. Despite the several crystal structures of Lys49-PLA2 homologues already determined, their biological assembly remains controversial with two possible conformations. The extended dimer with the hydrophobic channel exposed to the solvent and the compact dimer in which the active site-like region is occluded by the dimeric interface. In the MTX-II crystal packing analysis was found only the extended dimer as a possible stable quaternary arrangement. © 2012 Elsevier B.V.
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Snake venom serine proteinases (SVSPs) are hemostatically active toxins that perturb the maintenance and regulation of both the blood coagulation cascade and fibrinolytic feedback system at specific points, and hence, are widely used as tools in pharmacological and clinical diagnosis. The crystal structure of a thrombin-like enzyme (TLE) from Bothrops jararacussu venom (Jararacussin-I) was determined at 2.48 Å resolution. This is the first crystal structure of a TLE and allows structural comparisons with both the Agkistrodon contortrix contortrix Protein C Activator and the Trimeresurus stejnegeri plasminogen activator. Despite the highly conserved overall fold, significant differences in the amino acid compositions and three-dimensional conformations of the loops surrounding the active site significantly alter the molecular topography and charge distribution profile of the catalytic interface. In contrast to other SVSPs, the catalytic interface of Jararacussin-I is highly negatively charged, which contributes to its unique macromolecular selectivity. © 2012 The Protein Society.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Metagenomics has been widely employed for discovery of new enzymes and pathways to conversion of lignocellulosic biomass to fuels and chemicals. In this context, the present study reports the isolation, recombinant expression, biochemical and structural characterization of a novel endoxylanase family GH10 (SCXyl) identified from sugarcane soil metagenome. The recombinant SCXyl was highly active against xylan from beechwood and showed optimal enzyme activity at pH 6,0 and 45°C. The crystal structure was solved at 2.75 Å resolution, revealing the classical (β/α)8-barrel fold with a conserved active-site pocket and an inherent flexibility of the Trp281-Arg291 loop that can adopt distinct conformational states depending on substrate binding. The capillary electrophoresis analysis of degradation products evidenced that the enzyme displays unusual capacity to degrade small xylooligosaccharides, such as xylotriose, which is consistent to the hydrophobic contacts at the +1 subsite and low-binding energies of subsites that are distant from the site of hydrolysis. The main reaction products from xylan polymers and phosphoric acid-pretreated sugarcane bagasse (PASB) were xylooligosaccharides, but, after a longer incubation time, xylobiose and xylose were also formed. Moreover, the use of SCXyl as pre-treatment step of PASB, prior to the addition of commercial cellulolytic cocktail, significantly enhanced the saccharification process. All these characteristics demonstrate the advantageous application of this enzyme in several biotechnological processes in food and feed industry and also in the enzymatic pretreatment of biomass for feedstock and ethanol production. © 2013 Alvarez et al.