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.

Brewer's yeast and sugarcane yeast as protein sources for dogs

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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.

Energy Requirements for Maintenance and Growth of Male Saanen Goat Kids

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Energy Requirements in Early Life Are Similar for Male and Female Goat Kids

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

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)

Protein diets promote the maturation of oocytes and spawning of Piaractus mesopotamicus kept in cages

The objective of this study was to evaluate the effects of balanced diets on the maturation of oocytes and the reproductive performance of P. mesopotamicus in cages. A completely random design with 224 fish in 16 cages measuring 5 m(3) was employed for this purpose. The treatments consisted of diets containing 18, 24, 30, and 36% crude protein (CP) provided ad libitum. The external and internal morphological characteristics of the specimens were examined, as well as: the position of the germinal vesicle, the distribution of oocyte diameters, the fertilization and hatching rates, the number of oocytes released, the total number of oocytes, the remaining weight and total weight of the ovaries, the gonadosomatic index, the condition factor (K), and the histology of the oocytes and ovaries post-spawning and during ovarian regression. The diameters of the oocytes collected before the first hormonal application displayed a unimodal distribution for the lowest protein content and a polymodal distribution for the other treatments. A similar situation was seen during spawning. The lowest fertilization and hatching rates were found as a consequence of the treatment with 30% CP (P < 0.05). The greatest hatching rate occurred in the females fed 18% CP. The greatest total oocyte weight was found in the specimens that received between 30 and 36% CP. The lowest K index was found in the females fed 36% CP. In conclusion, a diet containing 18% CP satisfies the reproductive requirements of females adapted to this system.

Metabolizable energy and oil intake in brown commercial layers

With the objective to establish the best metabolizable energy (ME) intake for layers, and the best dietary vegetable oil addition level to optimize egg production, an experiment was carried out with 432 30-week-old Hisex Brown layers. Birds were distributed into nine treatments with six replicates of eight birds each according to a 3 x 3 factorial arrangement, consisting of three daily metabolizable energy intake (280, 300 or 320 kcal/bird/day) and three oil levels (0.00; 0.75 and 1.50 g/bird/day). Daily feed intake was limited to 115, 110 and 105 g/bird in order to obtain the desired energy and oil intake in each treatment. The following parameters were evaluated: initial weight, final weight, body weight change, egg production, egg mass, feed conversion ratio per dozen eggs and per egg mass and energy conversion. There was no influence of the treatments on egg production (%) or egg mass (g/bird/day). Final weight and body weight change were significantly affected by increasing energy intake. Feed conversion ratio per egg mass, feed conversion ratio per dozen eggs and energy conversion significantly worsened as a function of the increase in daily energy intake. An energy intake of 280 kcal/bird/day with no addition of dietary oil does not affect layer performance.

Synthesis, Biological Evaluation, And Molecular Modeling of Chalcone Derivatives As Potent Inhibitors of Mycobacterium tuberculosis Protein Tyrosine Phosphatases (PtpA and PtpB)

Tuberculosis (TB) is a major infectious disease caused by Mycobacterium tuberculosis (Mtb). According to the World Health Organization (WHO), about 1.8 million people die from TB and 10 million new cases are recorded each year. Recently, a new series of naphthylchalcones has been identified as inhibitors of Mtb protein tyrosine phosphatases (PTPs). In this work, 100 chalcones were designed, synthesized, and investigated for their inhibitory properties against MtbPtps. Structure-activity relationships (SAR) were developed, leading to the discovery of new potent inhibitors with IC50 values in the low-micromolar range. Kinetic studies revealed competitive inhibition and high selectivity toward the Mtb enzymes. Molecular modeling investigations were carried out with the aim of revealing the most relevant structural requirements underlying the binding affinity and selectivity of this series of inhibitors as potential anti-TB drugs.

Systematic studies of protein immobilization by surface plasmon field-enhanced fluorescence spectroscopy

The research interest of this study is to investigate surface immobilization strategies for proteins and other biomolecules by the surface plasmon field-enhanced fluorescence spectroscopy (SPFS) technique. The recrystallization features of the S-layer proteins and the possibility of combining the S-layer lattice arrays with other functional molecules make this protein a prime candidate for supramolecular architectures. The recrystallization behavior on gold or on the secondary cell wall polymer (SCWP) was recorded by SPR. The optical thicknesses and surface densities for different protein layers were calculated. In DNA hybridization tests performed in order to discriminate different mismatches, recombinant S-layer-streptavidin fusion protein matrices showed their potential for new microarrays. Moreover, SCWPs coated gold chips, covered with a controlled and oriented assembly of S-layer fusion proteins, represent an even more sensitive fluorescence testing platform. Additionally, S-layer fusion proteins as the matrix for LHCII immobilization strongly demonstrate superiority over routine approaches, proving the possibility of utilizing them as a new strategy for biomolecular coupling. In the study of the SPFS hCG immunoassay, the biophysical and immunological characteristics of this glycoprotein hormone were presented first. After the investigation of the effect of the biotin thiol dilution on the coupling efficiently, the interfacial binding model including the appropriate binary SAM structure and the versatile streptavidin-biotin interaction was chosen as the basic supramolecular architecture for the fabrication of a SPFS-based immunoassay. Next, the affinity characteristics between different antibodies and hCG were measured via an equilibrium binding analysis, which is the first example for the titration of such a high affinity interaction by SPFS. The results agree very well with the constants derived from the literature. Finally, a sandwich assay and a competitive assay were selected as templates for SPFS-based hCG detection, and an excellent LOD of 0.15 mIU/ml was attained via the “one step” sandwich method. Such high sensitivity not only fulfills clinical requirements, but is also better than most other biosensors. Fully understanding how LHCII complexes transfer the sunlight energy directionally and efficiently to the reaction center is potentially useful for constructing biomimetic devices as solar cells. After the introduction of the structural and the spectroscopic features of LHCII, different surface immobilization strategies of LHCII were summarized next. Among them the strategy based on the His-tag and the immobilized metal (ion) affinity chromatography (IMAC) technique were of great interest and resulted in different kinds of home-fabricated His-tag chelating chips. Their substantial protein coupling capacity, maintenance of high biological activity and a remarkably repeatable binding ability on the same chip after regeneration was demonstrated. Moreover, different parameters related to the stability of surface coupled reconstituted complexes, including sucrose, detergent, lipid, oligomerization, temperature and circulation rate, were evaluated in order to standardize the most effective immobilization conditions. In addition, partial lipid bilayers obtained from LHCII contained proteo-liposomes fusion on the surface were observed by the QCM technique. Finally, the inter-complex energy transfer between neighboring LHCIIs on a gold protected silver surface by excitation with a blue laser (λ = 473nm) was recorded for the first time, and the factors influencing the energy transfer efficiency were evaluated.

Distinct requirements for activation-induced cell surface expression of preformed Fas/CD95 ligand and cytolytic granule markers in T cells

Fas (CD95/Apo-1) ligand is a potent inducer of apoptosis and one of the major killing effector mechanisms of cytotoxic T cells. Thus, Fas ligand activity has to be tightly regulated, involving various transcriptional and post-transcriptional processes. For example, preformed Fas ligand is stored in secretory lysosomes of activated T cells, and rapidly released by degranulation upon reactivation. In this study, we analyzed the minimal requirements for activation-induced degranulation of Fas ligand. T cell receptor activation can be mimicked by calcium ionophore and phorbol ester. Unexpectedly, we found that stimulation with phorbol ester alone is sufficient to trigger Fas ligand release, whereas calcium ionophore is neither sufficient nor necessary. The relevance of this process was confirmed in primary CD4(+) and CD8(+) T cells and NK cells. Although the activation of protein kinase(s) was absolutely required for Fas ligand degranulation, protein kinase C or A were not involved. Previous reports have shown that preformed Fas ligand co-localizes with other markers of cytolytic granules. We found, however, that the activation-induced degranulation of Fas ligand has distinct requirements and involves different mechanisms than those of the granule markers CD63 and CD107a/Lamp-1. We conclude that activation-induced degranulation of Fas ligand in cytotoxic lymphocytes is differently regulated than other classical cytotoxic granule proteins.

Programmed Feeding of Protein to Finishing Beef Steers

Six-hundred pound Angus steer calves were fed cornbased finishing diets for 180 days to determine the effects of stepwise reduction of protein in the diet on performance and carcass characteristics. Reducing protein in the diet, but satisfying the requirements projected by the National Research Council model for Nutrient Requirements of Beef Cattle, did not affect performance or carcass measurements. Further reduction in protein content of the diet, so the projected requirement of the rumen microorganisms was not being met, did not affect performance or carcass measurements. It is concluded that quantity of protein fed to finishing cattle can be programmed and abstantially reduced. These reductions will result in substantially less nitrogen excreted in manure from larger feedlots.

Requirements for Staphylococcus aureus in vitro cellular internalization

Staphylcoccus aureus is a prokaryotic organism capable of causing numerous superficial and severe human infections. Adhesion of S. aureus to host tissues or cells is believed to be a crucial event in S. aureus infections. Subsequently, S. aureus can seed into the bloodstream resulting in metastasis of the infection. Several reports show that S. aureus can be internalized by non-professional phagocytes, a process which has been proposed to be important in S. aureus dissemination. An intracellular residence has also been proposed to provide safe harbor to reservoirs of dormant bacteria contributing to the persistence of infection. This dissertation describes an investigation into the molecular mechanisms of S. aureus internalization into both fibroblast and epithelial cells. Bacterial requirements for internalization were found to be limited to expression of proteins that bind the extracellular matrix protein fibronectin. A previously unknown fibronectin-binding region in the S. aureus fibronectin-binding protein A was discovered after showing competitive inhibition of S. aureus internalization. This novel fibronectin-binding activity is characterized. Internalization also required cell-based factors. The presence of fibronectin and cell surface receptors of the β1 integrin class, which are known to bind and internalize fibronectin, were found to be necessary for optimal internalization of S. aureus. These results led to the conclusion that fibronectin acts as a bridge between the bacterium and integrins on the host cells. The internalization process exhibits features characteristic of integrin-mediated cell migration on fibronectin-coated surfaces. Both processes involved an active form of the β1 integrin subunit and the protein tyrosine kinase Src. Finally, a Src inhibitor previously shown to be effective in reducing osteoporosis in an in vivo rat model is capable of greatly reducing S. aureus internalization. ^

Identifying the roles of the Escherichia coli FtsZ-associated proteins by minimizing the requirements for division

Formation of the FtsZ ring (Z ring) in Escherichia coli is the first step in assembly of the divisome, a molecular machine composed of 14 known proteins which are all required for cell division. Although the biochemical functions of most divisome proteins are unknown, several of these have overlapping roles in ensuring that the Z ring assembles at the cytoplasmic membrane and is active. ^ We identified a single amino acid change in FtsA, R286W, renamed FtsA*, that completely bypasses the requirement for ZipA in cell division. This and other data suggest that FtsA* is a hyperactive form of FtsA that can replace the multiple functions normally assumed by ZipA, which include stabilization of Z rings, recruitment of downstream cell division proteins, and anchoring the Z ring to the membrane. This is the first example of complete functional replacement of an essential prokaryotic cell division protein by another. ^ Cells expressing ftsA* with a complete deletion of ftsK are viable and divide, although many of these ftsK null cells formed multiseptate chains, suggesting a role in cell separation for FtsK. In addition, strains expressing extra ftsAZ, ftsQ, ftsB, zipA or ftsN, were also able to survive and divide in the absence of ftsK. The cytoplasmic and transmembrane domains of FtsQ were sufficient to allow viability and septum formation to ftsK deleted strains. These findings suggest that FtsK is normally involved in stabilizing the divisome and shares functional overlap with other cell division proteins. ^ As well as permitting the removal of other divisome components, the presence of FtsA* in otherwise wild-type cells accelerated Z-ring assembly, which resulted in a significant decrease in the average length of cells. In support of its role in Z-ring stability, FtsA* suppressed the cell division inhibition caused by overexpressing FtsZ. FtsA* did not affect FtsZ turnover within the Z ring as measured by fluorescence recovery after photobleaching. Turnover of FtsA* in the ring was somewhat faster than wild-type FtsA. Yeast two-hybrid data suggest that FtsA* has an increased affinity for FtsZ relative to wild-type FtsA. These results indicate that FtsA* interacts with FtsZ more strongly, and its enhancement of Z ring assembly may explain why FtsA* can permit survival of cells lacking ZipA or FtsK.^