Digestible tryptophan requirements for broilers from 22 to 42 days old

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

Digestible valine requirements for broilers from 22 and 42 days old

Current experiment established different criteria to evaluate the requirements of digestible valine for broilers from 22 and 42 days of age, by different regression models (quadratic, exponential and Linear Response Plateau) and, in the case of statistical significance, the comparison of means by Duncan test at 5% probability. A total of 1,920 Cobb 500 male broilers were used and distributed in an entirely randomized experimental design, with 6 treatments (6 digestible valine levels: 0.7192, 0.7729, 0.8265, 0.8802, 0.9339 and 0.9876%) and 8 replications, with 40 broilers each. Data on performance and carcass characteristics were evaluated. The level of 0.8265% digestible valine was considered standard. The inclusion of 0.816, 0.848 and 0.903% of digestible valine levels, corresponding to digestible valine:lysine ratios of approximately 76.00%, 79.00% and 84.12%, provided best feed intake, weight gain and feed conversion ratio, respectively for broiler from 22 to 42 days of age.

Modelling of the nitrogen deposition and dietary lysine requirements of Redbro broilers

The aim of this study was to determine the coefficients of the Goettingen model for Redbro birds and estimate the digestible lysine requirements. To determine the model parameters, three nitrogen balance trials were performed in Periods I (14-28 days), II (42-56 days) and III (70-84 days), using 42 birds per trial. The birds were individually housed and subjected to six diets with increasing levels of nitrogen, with lysine as the limiting amino acid (deficient by 20% in relation to other amino acids). Dietary nitrogen concentrations were 8, 16, 24, 32, 40 and 48 g/kg. A control diet was added to confirm lysine as the first limiting amino acid. Nitrogen balance trials were divided into 5 days of adaptation and two periods of excreta collection, each one of 5 days. The response of the birds to a control diet confirmed that lysine was the first limiting amino acid. The adjustment of the exponential functions between nitrogen retention or excretion and nitrogen intake allowed estimation of parameters of the Goettingen model. The maximum potential for nitrogen retention was 3276, 2585 and 2603 mg/BWkg0.67.day, nitrogen maintenance requirement was 225, 135 and 122 mg/BWkg0.67.day and efficiency of nitrogen utilisation was 313 x 10(-6), 406 x 10(-6) and 415 x 10(-6) in the phases of 14-28, 42-56 and 70-84 days. The digestible lysine intake for Periods I, II and III, based on 60% of the maximum potential for nitrogen retention, was 711, 989 and 1272 mg/day (1.225%, 1.137% and 1.09% of lysine in the diet for a daily feed intake of 58, 87 and 117 g/day), respectively.

Model to estimate lysine requirements of broilers

The purpose of this study is to describe the development of a model to predict the digestible lysine requirements of broilers using the factorial approach, and to evaluate the model using as reference the model presented in Brazilian Tables for Poultry and Swine. The model partitions the requirement for maintenance and growth for feather-free body protein and feather protein, in which the inputs are body and feather protein weight and the daily rates of protein deposition in the feather-free body and feathers. The parameters that express the lysine requirement for maintenance were obtained in metabolism trials with roosters, and those for the efficiency of lysine utilization in experiments with broilers from 1 to 42 d. Based on these results the model proposed was: Lys (mg/d) = [(151xBP(m)(-0.27)xBP(t)) + (0.01xP(t)x18)] + [(75xBPD/0.77) + (18xFPD/0.77)], where Lys = digestible lysine requirement (mg/d), BPm=body protein weight at maturity (kg), BPt=body protein weight at time t (kg), FPt=feather protein weight at time t (kg), BPD=body protein deposition (g/d), FPD = feather protein deposition (g/d). The model yields sensible predictions of the digestible lysine requirements of broilers of different strains and ages growing at their potential, and suggests a lower lysine requirement after 27 d than does the Brazilian model. The proposed model is the first step in the development of a simulation model that predicts the food intake of a broiler and hence the dietary amino acid content that would optimise performance.

Differences in larval nutritional requirements and female oviposition preference reflect the order of fruit colonization of Zaprionus indianus and Drosophila simulans

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Diet formulation techniques and lysine requirements of 1-to 22-day-old broilers

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

The Cysteine-Rich Protein Thimet Oligopeptidase as a Model of the Structural Requirements for S-glutathiolation and Oxidative Oligomerization

Thimet oligopeptidase (EP24.15) is a cysteine-rich metallopeptidase containing fifteen Cys residues and no intra-protein disulfide bonds. Previous work on this enzyme revealed that the oxidative oligomerization of EP24.15 is triggered by S-glutathiolation at physiological GSSG levels (10-50 mu M) via a mechanism based on thiol-disulfide exchange. In the present work, our aim was to identify EP24.15 Cys residues that are prone to S-glutathiolation and to determine which structural features in the cysteinyl bulk are responsible for the formation of mixed disulfides through the reaction with GSSG and, in this particular case, the Cys residues within EP24.15 that favor either S-glutathiolation or inter-protein thiol-disulfide exchange. These studies were conducted by in silico structural analyses and simulations as well as site-specific mutation. S-glutathiolation was determined by mass spectrometric analyses and western blotting with anti-glutathione antibody. The results indicated that the stabilization of a thiolate sulfhydryl and the solvent accessibility of the cysteines are necessary for S-thiolation. The Solvent Access Surface analysis of the Cys residues prone to glutathione modification showed that the S-glutathiolated Cys residues are located inside pockets where the sulfur atom comes into contact with the solvent and that the positively charged amino acids are directed toward these Cys residues. The simulation of a covalent glutathione docking onto the same Cys residues allowed for perfect glutathione posing. A mutation of the Arg residue 263 that forms a saline bridge to the Cys residue 175 significantly decreased the overall S-glutathiolation and oligomerization of EP24.15. The present results show for the first time the structural requirements for protein S-glutathiolation by GSSG and are consistent with our previous hypothesis that EP24.15 oligomerization is dependent on the electron transfer from specific protonated Cys residues of one molecule to previously S-glutathionylated Cys residues of another one.

Modificações nas frações que estimam carboidratos em plantas forrageiras com base nas equações da "Cornell Net Carbohydrate and Protein System"

Os organizadores autorizam a reprodução total ou parcial deste trabalho, para qualquer meio convencional ou eletrônico, para fins de estudo e pesquisa, desde que citada a fonte.

Determinants of the phagocytic signal mediated by the type IIIA Fc gamma receptor, Fc gamma RIIIA: sequence requirements and interaction with protein-tyrosine kinases.

The Fc gamma receptor-associated gamma and zeta subunits contain a conserved cytoplasmic motif, termed the immunoglobulin gene tyrosine activation motif, which contains a pair of YXXL sequences. The tyrosine residues within these YXXL sequences have been shown to be required for transduction of a phagocytic signal. We have previously reported that the gamma subunit of the type IIIA Fc gamma receptor (Fc gamma RIIIA) is approximately 6 times more efficient in mediating phagocytosis than the zeta subunit of Fc gamma RIIIA. By exchanging regions of the cytoplasmic domains of the homologous gamma and zeta chains, we observed that the cytoplasmic area of the gamma chain bearing a pair of the conserved YXXL sequences is important in phagocytic signaling. Further specificity of phagocytic signaling is largely determined by the two internal XX amino acids in the YXXL sequences. In contrast, the flanking amino acids of the YXXL sequences including the seven intervening amino acids between the two YXXL sequences do not significantly affect the phagocytic signal. Furthermore, the protein-tyrosine kinase Syk, but not the related kinase ZAP-70, stimulated Fc gamma RIIIA-mediated phagocytosis. ZAP-70, however, increased phagocytosis when coexpressed with the Src family kinase Fyn. These data demonstrate the importance of the two specific amino acids within the gamma subunit YXXL cytoplasmic sequences in phagocytic signaling and explain the difference in phagocytic efficiency of the gamma and zeta chains. These results indicate the importance of Syk in Fc gamma RIIIA-mediated phagocytosis and demonstrate that ZAP-70 and syk differ in their requirement for a Src-related kinase in signal transduction.

Differences in larval nutritional requirements and female oviposition preference reflect the order of fruit colonization of Zaprionus indianus and Drosophila simulans

Species coexist using the same nutritional resource by partitioning it either in space or time, but few studies explore how species-specific nutritional requirements allow partitioning. Zaprionus indianus and Drosophila simulans co-exist in figs by invading the fruit at different stages; Z. indianus colonizes ripe figs, whereas D. simulans oviposits in decaying fruit. Larvae feed on yeast growing on the fruit, which serves as their primary protein source. Because yeast populations increase as fruit decays, we find that ripe fruit has lower protein content than rotting fruit. Therefore, we hypothesized that Z. indianus and D. simulans larvae differ in their dietary requirements for protein. We used nutritional geometry to assess the effects of protein and carbohydrate concentration in the larval diet on life history characters in both species. Survival, development time, and ovariole number respond differently to the composition of the larval diet, with Z. indianus generally performing better across a wider range of protein concentrations. Correspondingly, we found that Z. indianus females preferred to lay eggs on low protein foods, while D. simulans females chose higher protein foods for oviposition when competing with Z. indianus. We propose the different nutritional requirements and oviposition preference of these two species allows them to temporally partition their habitat.

Studies of Protein-Protein and Protein-Water Interactions by Small Angle X-Ray Scattering, Terahertz Spectroscopy, ASMOS, And Computer Simulation

The protein folding problem has been one of the most challenging subjects in biological physics due to its complexity. Energy landscape theory based on statistical mechanics provides a thermodynamic interpretation of the protein folding process. We have been working to answer fundamental questions about protein-protein and protein-water interactions, which are very important for describing the energy landscape surface of proteins correctly. At first, we present a new method for computing protein-protein interaction potentials of solvated proteins directly from SAXS data. An ensemble of proteins was modeled by Metropolis Monte Carlo and Molecular Dynamics simulations, and the global X-ray scattering of the whole model ensemble was computed at each snapshot of the simulation. The interaction potential model was optimized and iterated by a Levenberg-Marquardt algorithm. Secondly, we report that terahertz spectroscopy directly probes hydration dynamics around proteins and determines the size of the dynamical hydration shell. We also present the sequence and pH-dependence of the hydration shell and the effect of the hydrophobicity. On the other hand, kinetic terahertz absorption (KITA) spectroscopy is introduced to study the refolding kinetics of ubiquitin and its mutants. KITA results are compared to small angle X-ray scattering, tryptophan fluorescence, and circular dichroism results. We propose that KITA monitors the rearrangement of hydrogen bonding during secondary structure formation. Finally, we present development of the automated single molecule operating system (ASMOS) for a high throughput single molecule detector, which levitates a single protein molecule in a 10 µm diameter droplet by the laser guidance. I also have performed supporting calculations and simulations with my own program codes.

Proteasome Inhibition Represses Unfolded Protein Response and Nox4, Sensitizing Vascular Cells to Endoplasmic Reticulum Stress-Induced Death

Background: Endoplasmic reticulum (ER) stress has pathophysiological relevance in vascular diseases and merges with proteasome function. Proteasome inhibition induces cell stress and may have therapeutic implications. However, whether proteasome inhibition potentiates ER stress-induced apoptosis and the possible mechanisms involved in this process are unclear. Methodology/Principal Findings: Here we show that proteasome inhibition with MG132, per se at non-lethal levels, sensitized vascular smooth muscle cells to caspase-3 activation and cell death during ER stress induced by tunicamycin (Tn). This effect was accompanied by suppression of both proadaptive (KDEL chaperones) and proapoptotic (CHOP/GADD153) unfolded protein response markers, although, intriguingly, the splicing of XBP1 was markedly enhanced and sustained. In parallel, proteasome inhibition completely prevented ER stress-induced increase in NADPH oxidase activity, as well as increases in Nox4 isoform and protein disulfide isomerase mRNA expression. Increased Akt phosphorylation due to proteasome inhibition partially offset the proapoptotic effect of Tn or MG132. Although proteasome inhibition enhanced oxidative stress, reactive oxygen species scavenging had no net effect on sensitization to Tn or MG132-induced cell death. Conclusion/Relevance: These data indicate unfolded protein response-independent pathways whereby proteasome inhibition sensitizes vascular smooth muscle to ER stress-mediated cell death. This may be relevant to understand the therapeutic potential of such compounds in vascular disease associated with increased neointimal hyperplasia.

Angiotensin receptor blockade improves the net balance of cardiac Ca(2+) handling-related proteins in sympathetic hyperactivity-induced heart failure

Aims: The clinical benefits of angiotensin II type 1 (AT1) receptor blockers (ARB) in heart failure (HF) include cardiac anti-remodeling and improved ventricular function. However, the cellular mechanisms underlying the benefits of ARB on ventricular function need to be better clarified. In the present manuscript, we evaluated the effects of AT1 receptor blockade on the net balance of Ca(2+) handling proteins in hearts of mice lacking alpha(2A) and alpha(2C) adrenoceptors (alpha(2A)/alpha(2C)ARKO), which develop sympathetic hyperactivity (SH) induced-HF. Main methods: A cohort of male wild-type (WT) and congenic alpha(2A)/alpha(2C)ARKO mice in a C57BL6/J genetic background (5-7 mo of age) was randomly assigned to receive either placebo or ARB (Losartan, 10 mg/kg for 8wks). Ventricular function (VF) was assessed by echocardiography, and cardiac myocyte width and ventricular fibrosis by a computer-assisted morphometric system. Sarcoplasmic reticulum Ca(2+) ATPase (SERCA2), phospholamban (PLN), phospho-Ser(16)-PLN, phospho-Thr(17)-PLN, phosphatase 1 (PP1), Na(+)-Ca(2+) exchanger (NCX), Ca(2+)/calmodulin-dependent protein kinase 11 (CaMKII) and phospho-Thr(286)-CaMKII were analyzed by Western blot. Key findings: alpha(2A)/alpha(2C)ARKO mice displayed ventricular dysfunction, cardiomyocyte hypertrophy and cardiac fibrosis paralleled by decreased SERCA2 and increased phospho-Thr(17)-PLN, CaMKII, phospho-Thr(286)-CaMKII and NCX levels. ARB induced anti-cardiac remodeling effect and improved VF in alpha(2A)/alpha(2C)ARKO associated with increased SERCA2 and phospho-Ser(16)-PLN levels, and SERCA2:NCX ratio. Additionally, ARB decreased phospho-Thr(17)-PLN levels as well as reestablished NCX, CaMKII and phospho-Thr(286)-CaMKII toward WT levels. Significance: Altogether, these data provide new insights on intracellular Ca(2+) regulatory mechanisms underlying improved ventricular function by ARB therapy in HF. (c) 2011 Elsevier Inc. All rights reserved.