INTERCROPPING CORN WITH A COMBINATION OF TREE SPECIES TO CONTROL WEEDS

ABSTRACT The combination of crop residues or crop extracts is often more advantageous in controlling weeds, than the application of each residue or extract singly. This suggests that in intercropping with maize, the combination of tree species can be more advantageous than species isolated in weed control. The objective of this study was to evaluate the effects of intercropping with a combination of leguminous on the weed growth and corn yield. A randomized-block design with split plots (cultivars in plots) and five replicates was established. The cultivars BR 205 and AG 1041 were subject to the following treatments: two weedings (A), intercropping with sabiá (B), gliricidia (C), gliricidia + sabiá (D) and no weeding (E). In the B and C, 30 viable seeds m-2 of the leguminous were sown. In the D, 15 seeds of each species were sown m-2. The legumes were sown by random casting during corn planting. The sequence of the best treatments in reducing the growth of weeds is A > B = C = D = E. The sequence of the best treatments when are considered the yields of baby corn, green corn and grain is A > B > C > D > E. The cultivars do not differ in regards to the reduction in weed growth. In terms of corn yield cultivar BR 205 is the best.

Allelopathic Influence of Residues from Sphagneticola trilobata on Weeds and Crops

The allelopathic effect studied in many cultures has currently generated great expectations that displayed a natural and environmentally friendly tool for weed management using bioherbicides. The objective of this work was to assess allelopathic influence of residues of S. trilobata on the germination and growth of weeds, as well as their relation with some crops and effects on soil properties. Results show that residues from S. trilobata have inhibited the germination of weeds (31.6 - 72%), increasingly with the applied dose. All residue doses of this specie have inhibited dicotyledonous germination, but only maximum concentration has affected monocotyledons. The residues did not affect onion germination, but stimulated it in radish and tomato, while the dose applied at 50% produced tomato stimulation and inhibition of cabbage. The effects of residues on hypocotyl growth in different crops showed changes in species response. For onion, the three doses had negative effects on the growth of hypocotyl, while tomato was stimulated. For radish, the growth was hindered by any dose applied, and were only different (50 and 100%) compared to control. For cabbage, only hypocotyl length was stimulated, when maximum dose (100%) was applied. For the radicle growth, in onion and radish no differences were found compared to control. While the tomato radicle growth was inhibited, in cabbage, all doses encouraged the elongation of the radicle. The dry mass of weed was affected by increased dose of residue (0.49 - 8.8 g m-2), however the soil microflora was stimulated, while the population of Azotobacter spp. was not affect. Some soil properties were affected, the level of organic material, Na+ and electrical conductivity were increased, while pH (H2O) decreased a bit, however it remained basic.

Eucalypt Growth Submitted to Management of Urochloa spp.

The objective of this work was to evaluate the initial growth and the physiological characteristics of eucalypt submitted to different managements of signalgrass. The experiment was conducted in a protected environment, using a randomized blocks design with five repetitions. The treatments were arranged in a (5 x 2) + 1 factorial design, and the first factor corresponded to the types of weed management (no control; chemical control keeping the signalgrass shoot on the soil; chemical control with removal of the signalgrass shoot; mechanical control keeping the signalgrass shoot on the soil, and mechanical control with removal of the signalgrass shoot); the second factor corresponded to the two weeds species (U. brizantha and U. decumbens) and a control relative to the eucalypt in monoculture. The eucalypt growth was not affected by the presence of the Urochloa species until 50 days after treatments (DAT). However, the coexistence of these species with eucalypt for 107 DAT reduced the collar diameter, total dry matter, and the leaf area, but did not alter the characteristics related to photosynthesis and transpiration. The control method adopted, with removal or maintenance of the signalgrass shoot, regardless of species, did not change the initial eucalypt growth. It can be concluded that the coexistence of eucalypt with Urochloa decumbens or Urochloa brizantha for 105 days reduces the eucalypt growth. However, the use of chemical or mechanical control, with or without removal of signalgrass residue, were effective methods to prevent interference of these weeds.

Flavianate, an amino acid precipitant, is a competitive inhibitor of trypsin at pH 3.0

Textile dyes bind to proteins leading to selective co-precipitation of a complex involving one protein molecule and more than one dye molecule of opposite charge in acid solutions, in a process of reversible denaturation that can be utilized for protein fractionation. In order to understand what occurs before the co-precipitation, a kinetic study using bovine ß-trypsin and sodium flavianate was carried out based on reaction progress curve techniques. The experiments were carried out using a-CBZ-L-Lys-p-nitrophenyl ester as substrate which was added to 50 mM sodium citrate buffer, pH 3.0, containing varying concentrations of ß-trypsin and dye. The reaction was recorded spectrophotometrically at 340 nm for 30 min, and the families of curves obtained were analyzed simultaneously by fitting integrated Michaelis-Menten equations. The dye used behaved as a competitive inhibitor of trypsin at pH 3.0, with Ki = 99 µM; kinetic parameters for the substrate hydrolysis were: Km = 32 µM, and kcat = 0.38/min. The competitive character of the inhibition suggests a specific binding of the first dye molecule to His-57, the only positively charged residue at the active site of the enzyme.

Development of an operational substrate for ZapA, a metalloprotease secreted by the bacterium Proteus mirabilis

The protease ZapA, secreted by Proteus mirabilis, has been considered to be a virulence factor of this opportunistic bacterium. The control of its expression requires the use of an appropriate methodology, which until now has not been developed. The present study focused on the replacement of azocasein with fluorogenic substrates, and on the definition of enzyme specificity. Eight fluorogenic substrates were tested, and the peptide Abz-Ala-Phe-Arg-Ser-Ala-Ala-Gln-EDDnp was found to be the most convenient for use as an operational substrate for ZapA. A single peptide bond (Arg-Ser) was cleaved with a Km of 4.6 µM, a k cat of 1.73 s-1, and a catalytic efficiency of 376 (mM s)-1. Another good substrate for ZapA was peptide 6 (Abz-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-Gln-EDDnp) which was cleaved at a single bond (Phe-Ser) with a Km of 13.6 µM, a k cat of 3.96 s-1 and a catalytic efficiency of 291 (mM s)-1. The properties of the amino acids flanking the scissile bonds were also evaluated, and no clear requirement for the amino acid residue at P1 was found, although the enzyme seems to have a preference for a hydrophobic residue at P2.

Reaction of diphenyl diselenide with hydrogen peroxide and inhibition of delta-aminolevulinate dehydratase from rat liver and cucumber leaves

The interaction of the product of H2O2 and (PhSe)2 with delta-aminolevulinate dehydratase (delta-ALA-D) from mammals and plants was investigated. (PhSe)2 inhibited rat hepatic delta-ALA-D with an IC50 of 10 µM but not the enzyme from cucumber leaves. The reaction of (PhSe)2 with H2O2 for 1 h increased the inhibitory potency of the original compound and the IC50 for animal delta-ALA-D inhibition was decreased from 10 to 2 µM. delta-ALA-D from cucumber leaves was also inhibited by the products of reaction of (PhSe)2 with H2O2 with an IC50 of 4 µM. The major product of reaction of (PhSe)2 with H2O2 was identified as seleninic acid and produced an intermediate with a lambdamax at 265 nm after reaction with t-BuSH. These results suggest that the interaction of (PhSe)2 with mammal delta-ALA-D requires the presence of cysteinyl residues in close proximity. Two cysteine residues in spatial proximity have been recently described for the mammalian enzyme. Analysis of the primary structure of plant delta-ALA-D did not reveal an analogous site. In contrast to (PhSe)2, seleninic acid, as a result of the higher electrophilic nature of its selenium atom, may react with additional cysteinyl residue(s) in mammalian delta-ALA-D and also with cysteinyl residues from cucumber leaves located at a site distinct from that found at the B and A sites in mammals. Although the interaction of organochalcogens with H2O2 may have some antioxidant properties, the formation of seleninic acid as a product of this reaction may increase the toxicity of organic chalcogens such as (PhSe)2.

Hemodynamic and hormonal actions of adrenomedullin

Adrenomedullin, a 52-amino acid residue peptide, has numerous biological actions which are of potential importance to cardiovascular homeostasis, growth and development of cardiovascular tissues and bone, prevention of infection, and regulation of body fluid and electrolyte balance. Studies in man using intravenous infusion of the peptide have demonstrated that, at plasma levels detected after myocardial infarction or in heart failure, adrenomedullin reduces arterial pressure, increases heart rate and cardiac output, and activates the sympathetic and renin-angiotensin systems but suppresses aldosterone. The thresholds for these responses differ, being lower under some experimental circumstances for arterial pressure than for the other biological effects. Adrenomedullin administration inhibits the pressor and aldosterone-stimulating action of angiotensin II in man. By contrast, the pressor effect of norepinephrine is little altered by concomitant adrenomedullin administration. Although in the absence of a safe, specific antagonist of the actions of endogenous adrenomedullin it is difficult to be certain about the physiological and pathophysiological importance of this peptide in man, current evidence suggests that it serves to protect against cardiovascular overload and injury. Hope has been expressed that adrenomedullin or an agonist specific for adrenomedullin receptors might find a place in the treatment of cardiovascular disorders.

Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis

Lactococcus lactis, the model lactic acid bacterium, is a good candidate for heterologous protein production in both foodstuffs and the digestive tract. We attempted to produce Streptomyces tendae antifungal protein 1 (Afp1) in L. lactis with the objective of constructing a strain able to limit fungal growth. Since Afp1 activity requires disulfide bond (DSB) formation and since intracellular redox conditions are reportedly unfavorable for DSB formation in prokaryotes, Afp1 was produced as a secreted form. An inducible expression-secretion system was used to drive Afp1 secretion by L. lactis; Afp1 was fused or not with LEISSTCDA, a synthetic propeptide (LEISS) that has been described to be a secretion enhancer. Production of Afp1 alone was not achieved, but production of LEISS-Afp1 was confirmed by Western blot and immunodetection with anti-Afp1 antibodies. This protein (molecular mass: 9.8 kDa) is the smallest non-bacteriocin heterologous protein ever reported to be secreted in L. lactis via the Sec-dependent pathway. However, no anti-fungal activity was detected, even in concentrated samples of induced supernatant. This could be due to a too low secretion yield of Afp1 in L. lactis, to the absence of DSB formation, or to an improper DSB formation involving the additional cysteine residue included in LEISS propeptide. This raises questions about size limits, conformation problems, and protein secretion yields in L. lactis.

Unraveling the functional divergence of membrane-bound pyrophosphatases

Inorganic pyrophosphatases (PPases) are enzymes that hydrolyze pyrophosphate (PPi)which is produced as a byproduct in many important growth related processes e.g. in the biosynthesis of DNA, proteins and lipids. PPases can be either soluble or membranebound. Membrane-bound PPases (mPPases) are ion transporters that couple the energy released during PPi hydrolysis to Na+ or H+ transport. When I started the project, only three Na+-transporting mPPases were known to exist. In this study, I aimed to confirm if Na+-transport is a common function of mPPases. Furthermore, the amino acid residues responsible for determining the transporter specificity were unknown. I constructed a phylogenetic tree for mPPases and selected the representative bacterial and archaeal mPPases to be investigated. I expressed different prokaryotic mPPases in Escherichia coli, isolated these as inverted membrane vesicles and characterized their functions. In the first project I identified four new Na+-PPases, two K+-dependent H+-PPases and one K+-independent mPPase. The residues determining the transporter specificity were identified by site-directed mutagenesis. I showed that the conserved glutamate residues are important for specificity, though are not the only residues that influence it. This research clarified the ion transport specificities throughout the mPPase phylogenetic tree, and revealed that Na+ transport is a widespread function of mPPases. In addition, it became clear that the transporter specificity can be predicted from the amino acid sequence in combination with a phylogenetic analysis. In the second project, I identified a novel class of mPPases, which is capable of transporting both Na+ and H+ ions and is mainly found in bacteria of the human gastrointestinal tract. The physiological role of these novel enzymes may be to help the bacteria survive in the demanding conditions of the host. In the third project, I characterized the Chlorobium limicola Na+-PPase and found that this and related mPPases are able to transport H+ ions at subphysiological Na+ concentrations. In addition, the H+-transport activity was shown to be a common function of all studied Na+-PPases at low Na+ concentrations. I observed that mutating gate-lysine to asparagine eliminated the H+ but not the Na+ ion transport function, indicating the important role of the residue in the transport of H+. In the fourth project, I characterized the unknown and evolutionary divergent mPPase clade of the phylogenetic tree. The enzymes belonging to this clade are able to transport H+ ions and, based on their sequence, were expected to be K+- and Na+-independent. The sequences of membrane-bound PPase are usually highly conserved, but the enzymes belonging to this clade are more divergent and usually contain 100−150 extra amino acid residues compared to other known mPPases. Despite the vast sequence differences, these mPPases have the full set of important residues and, surprisingly, are regulated by Na+ and K+ ions. These enzymes are mainly of bacterial origin.

Characterization and oxygen binding properties of des-Arg human hemoglobin

The role of chloride in the stabilization of the deoxy conformation of hemoglobin (Hb), the low oxygen affinity state, has been studied in order to identify the nature of this binding. Previous studies have shown that arginines 141α could be involved in the binding of this ion to the protein. Thus, des-Arg Hb, human hemoglobin modified by removal of the α-chain C-terminal residue Arg141α, is a possible model for studies of these interactions. The loss of Arg141α and all the salt bridges in which it participates is associated with subtle structural perturbations of the α-chains, which include an increase in the conformational flexibility and further shift to the oxy state, increasing oxygen affinity. Thus, this Hb has been the target of many studies of structural and functional behavior along with medical applications. In the present study, we describe the biochemical characterization of des-Arg Hb by electrophoresis, high-performance liquid chromatography and mass spectroscopy. The effects of chloride binding on the oxygen affinity and on the cooperativity to des-Arg Hb and to native human hemoglobin, HbA, were measured and compared. We confirm that des-Arg Hb presents high oxygen affinity and low cooperativity in the presence of bound chloride and show that the binding of chloride to des-Arg does not change its functional characteristics as observed with HbA. These results indicate that Arg141α may be involved in the chloride effect on Hb oxygenation. Moreover, they show that these residues contribute to lower Hb oxygen affinity to a level compatible with its biological function.

The novel p.E89K mutation in the SRY gene inhibits DNA binding and causes the 46,XY disorder of sex development

Male sex determination in humans is controlled by the SRY gene, which encodes a transcriptional regulator containing a conserved high mobility group box domain (HMG-box) required for DNA binding. Mutations in the SRY HMG-box affect protein function, causing sex reversal phenotypes. In the present study, we describe a 19-year-old female presenting 46,XY karyotype with hypogonadism and primary amenorrhea that led to the diagnosis of 46,XY complete gonadal dysgenesis. The novel p.E89K missense mutation in the SRY HMG-box was identified as a de novo mutation. Electrophoretic mobility shift assays showed that p.E89K almost completely abolished SRY DNA-binding activity, suggesting that it is the cause of SRY function impairment. In addition, we report the occurrence of the p.G95R mutation in a 46,XY female with complete gonadal dysgenesis. According to the three-dimensional structure of the human SRY HMG-box, the substitution of the conserved glutamic acid residue by the basic lysine at position 89 introduces an extra positive charge adjacent to and between the positively charged residues R86 and K92, important for stabilizing the HMG-box helix 2 with DNA. Thus, we propose that an electrostatic repulsion caused by the proximity of these positive charges could destabilize the tip of helix 2, abrogating DNA interaction.

Nutritional aspects of nile tilapia (Oreochromis niloticus) silage

One third of the world's fishing produce is not directly used for human consumption. Instead, it is used for making animal food or is wasted as residue. It would be ideal to use the raw material thoroughly and to recover by-products, preventing the generation of residues. With the objectives of increasing the income and the production of the industry, as well as minimizing environmental and health problems from fish residue, chemical silage from Tilapia (Oreochromis niloticus) processing residues was developed after homogenization and acidification of the biomass with 3% formic acid: propionic, 1:1, addition of antioxidant BHT and maintenance of pH at approximately 4.0. Analyses to determine the moisture, protein, lipids and ash were carried out. The amino acids were examined in an auto analyzer after acid hydrolysis, except for the tryptophan which was determined through colorimetry. The tilapia silage presented contents that were similar to or higher than the FAO standards for all essential amino acids, except for the tryptophan. The highest values found were for glutamic acid, lysine and leucine. The results indicate a potential use of the silage prepared from the Nile tilapia processing residue as a protein source in the manufacturing of fish food.

Evaluation of the quality of a molded sweet biscuit enriched with okara flour

The objective of this research was to test the addition of soymilk residue, also known as okara, to a molded sweet biscuit (MSB). The okara was provided by two soymilk producing companies whose production systems are based on hot disintegration of decorticated (company B) or non-decorticated (company A) soybeans and separation of the soymilk. Okaras A and B were dehydrated in a flash dryer and then ground to a flour (< 200 mesh). The okara flours showed high protein (35 g.100 g-1 dwb), lipid (17 g.100 g-1 dwb), and fiber (17 to 21 g.100 g-1 dwb) contents. The water holding capacity, protein solubility, emulsifying capacity, emulsion stability and isoflavone contents found in flour A were significantly higher (p < 0.05) than in flour B. The formulation of MSB, replacing 30% (w/w) of the wheat flour with okara flour was tested. The results of the physical measurements, brittleness and water activity of the MSB with flours A and B did not differ significantly (p < 0.05) from those of the standard. The color, flavor and overall quality of the MSB with 30% of okara flour B did not differ significantly from those of the standard biscuit, demonstrating its potential for application in confectionery products.

Physical, chemical, technological and sensory characteristics of Frankfurter type sausage containing okara flour

The addition of okara flour to an emulsified meat product (Frankfurter type sausage) was evaluated based on the physical, chemical, technological, and sensory characteristics of the final product. Okara, residue from soymilk production, was provided by two soymilk producing companies whose production systems were based on the hot disintegration of the decorticated (company B) or undecorticated (company A) soybeans. The okara was dehydrated using a flash dryer and then ground into flour (>420 µm). However, The okara flours A and B showed approximately the same amount of protein (35 and 40 g.100 g-1 dwb). However, the okara flour A presented higher values (p < 0.05) for all technological functional properties studied (emulsification capacity, emulsion stability, protein solubility, and water hold capacity) than those of okara flour B. The A and B okara flours were used in a frankfurter sausage formulation as substitution of 1.5% and 4% of meat. The results showed that the sausages containing okara flours A and B, as well as the control sausage, were accepted by the sensory panel. Moreover, there were no significant differences (p < 0.05) in the physical (color, objective texture, and emulsion stability) and chemical (pH and proximate composition) measurements of the sausages with and without the okara flour.

Influence of process parameters on the color and texture of passion fruit albedo preserved in syrup

The industrialization of passion fruit in the form of juice produces considerable amounts of residue that could be used as food. The objective of the present study was to determine the effects of the volume of passion fruit juice added to the syrup and the cooking time on the color and texture of passion fruit albedo preserved in syrup. Multi-linear models were well fit to describe the value for a* (for the albedo) the values for b* (for the albedo and syrup), which exhibited high correlation coefficients of 98%, 84%, and 88%, respectively. The volume of passion fruit juice added and the cooking time of the albedos in the syrup, involved in the processing of passion fruit albedo preserves in syrup, significantly affected color analyses. The texture was not affected by the parameters studied. Therefore, the use of larger volumes of passion fruit juice and longer cooking time is recommended for the production of passion fruit albedo preserves in syrup to achieve the characteristic yellow color of the fruit.