Children's use of looking behavior as a cue to detect another's goal

Three studies examined children's understanding of the role that looking behavior plays in revealing another's desired goal. In each study, participants were asked which of 2 objects a protagonist wanted to obtain. Four-year-olds did not infer that an object examined via prolonged looking was more likely to be the protagonist's goal than an object that was either glanced at or inadvertently touched. Instead, they were accurate only when the protagonist looked at one of two potential goals. In contrast, the majority of 6-year-olds (and adults in Experiment 1) consistently regarded prolonged looking as the more important cue of the protagonist's goal. These age differences suggest that development is characterized by an increasing appreciation that goal is revealed by comparative differences in the quality of perceptual connectedness to objects in the world. One explanation for these age differences is that preschoolers are limited in their understanding of the difference between perceiving with full attention and without it.     

Ca2+ differentially regulates conventional protein kinase Cs' membrane interaction and activation

The regulation of conventional protein kinase Cs by Ca2+ was examined by determining how this cation affects the enzyme's 1) membrane binding and catalytic function and 2) conformation. In the first part, we show that significantly lower concentrations of Ca2+ are required to effect half-maximal membrane binding than to half-maximally activate the enzyme. The disparity between binding and activation kinetics is most striking for protein kinase C betaII, where the concentration of Ca2+ promoting half-maximal membrane binding is approximately 40-fold higher than the apparent Km for Ca2+ for activation. In addition, the Ca2+ requirement for activation of protein kinase C betaII is an order of magnitude greater than that for the alternatively spliced protein kinase C betaI; these isozymes differ only in 50 amino acids at the carboxyl terminus, revealing that residues in the carboxyl terminus influence the enzyme's Ca2+ regulation. In the second part, we use proteases as conformational probes to show that Ca2+dependent membrane binding and Ca2+-dependent activation involve two distinct sets of structural changes in protein kinase C betaII. Three separate domains spanning the entire protein participate in these conformational changes, suggesting significant interdomain interactions. A highly localized hinge motion between the regulatory and catalytic halves of the protein accompanies membrane binding; release of the carboxyl terminus accompanies the low affinity membrane binding mediated by concentrations of Ca2+ too low to promote catalysis; and exposure of the amino-terminal pseudosubstrate and masking of the carboxyl terminus accompany catalysis. In summary, these data reveal that structural determinants unique to each isozyme of protein kinase C dictate the enzyme's Ca2+-dependent affinity for acidic membranes and show that, surprisingly, some of these determinants are in the carboxyl terminus of the enzyme, distal from the Ca2+-binding site in the amino-terminal regulatory domain.     

Stabilization of apoflavodoxin by replacing hydrogen-bonded charged Asp or Glu residues by the neutral isosteric Asn or Gln

Knowledge of protein stability principles provides a means toincrease protein stability in a rational way. Here we explorethe feasibility of stabilizing proteins by replacing solvent-exposedhydrogen-bonded charged Asp or Glu residues by the neutral isostericAsn or Gln. The rationale behind this is a previous observationthat, in some cases, neutral hydrogen bonds may be more stablethat charged ones. We identified, in the apoflavodoxin fromAnabaena PCC 7119, three surface-exposed aspartate or glutamateresidues involved in hydrogen bonding with a single partnerand we mutated them to asparagine or glutamine, respectively.The effect of the mutations on apoflavodoxin stability was measuredby both urea and temperature denaturation. We observed thatthe three mutant proteins are more stable than wild-type (onaverage 0.43 kcal/mol from urea denaturation and 2.8°C froma two-state analysis of fluorescence thermal unfolding data).At high ionic strength, where potential electrostatic repulsionsin the acidic apoflavodoxin should be masked, the three mutantsare similarly more stable (on average 0.46 kcal/mol). To ruleout further that the stabilization observed is due to removalof electrostatic repulsions in apoflavodoxin upon mutation,we analysed three control mutants and showed that, when thecharged residue mutated to a neutral one is not hydrogen bonded,there is no general stabilizing effect. Replacing hydrogen-bondedcharged Asp or Glu residues by Asn or Gln, respectively, couldbe a straightforward strategy to increase protein stability.     

Synthesis,characterization and compared reactivity of asymmetrical ansa-metallocenes

Rac and meso ansa-zirconocenes [Zr{1-Me₂Si(3-R-(η⁵-C₉H₅))(3-R′-(η⁵-C₉H₅))}Cl₂] (R = Et, R′ = H; R = Pr, R′ = H; and R = Et, R′ = Pr) 4–9 have been prepared by the reaction of ZrCl₄ with the corresponding dilithiated derivatives from the ligands {1-Me₂Si(3-R-(η⁵-C₉H₅))(3-R′-(η⁵-C₉H₅))} (R = Et, R′ = H 1; R = Pr, R′ = H 2; and R = Et, R′ = Pr 3) in diethyl ether/toluene at ?78 °C. The molecular structure of rac [Zr{1-Me₂Si(3-Et-(η⁵-C₉H₅))(3-Pr-(η⁵-C₉H₅))}Cl₂] 8 has been determined by single crystal X-ray diffraction studies, which show a pseudotetrahedral environment formed by the two chlorine ligands and two η⁵-coordinated indenyl ligands. The activity in homogeneous and heterogeneous polymerization is compared and discussed.     

A review of natural antioxidants and their effects on oxidative status, odor and quality of fresh beef produced in Argentina

Meat derived from pasture feeding, is associated with a high level of antioxidants. Antioxidants are incorporated within cell membranes and protect tissues against oxidation from reactive oxygen species. This maintains the overall quality of meat and secondary products. This paper reviews the implications of incorporating natural antioxidants into fresh beef, focusing on the benefits of feeding cattle good quality pasture. Pasture samples typically have higher levels of α-tocopherol, β-carotene, ascorbic acid and glutathione than feedlot samples. These compounds retard lipid and protein oxidation in fresh and stored meat, and preserve the color and odor quality of beef. The significance of antioxidant enzymes is variable, because their behavior depends on individual redox status before slaughter. Understanding total antioxidant activity requires information on antioxidant and pro-oxidant status. With an abundance of pasture, Argentina has a natural advantage in producing meat with a high antioxidant value.     

Differentiation of intermediate products (concentrates and purées) from the fruit industry by means of phenolic content

Concentrates and purées are intermediate products in the manufacture of commercial fruit juices. In this paper, the phenolic content [total polyphenols (TP), ortho-diphenols (ORT) and catechins (CAT)] in a large number of concentrates and purées from apple and peach fruits has been determined. The relationships TP/ORT, TP/CAT and ORT/CAT have also been calculated. TP content was found to be significantly higher in the concentrates than in the respective purées whereas CAT content was significantly higher in the purée. ORT was found to be higher in purées than in apple concentrates, but the opposite was observed for peaches. The relationships TP/ORT, TP/CAT and ORT/CAT were higher for the concentrates than for the respective purées. In addition, values for ORT/CAT were higher for the peach products than for the respective apple products. The results show that these phenolic compound measurements enable differentiation between concentrates and purées, as well as between the fruit juices produced from them. The main advantage of measurements of total phenolic compounds is that they are easy to perform in a conventional laboratory, which makes them suitable for routine analysis.     

Viral pollution determination of superficial waters (river water and sea-water) from the urban area of Barcelona (Spain)

Various surface water systems in the metropolitan area of Barcelona (Spain), including two rivers and three beaches, were analysed for human viruses using glass powder as adsorbent to concentrate them. The method used allows concentration of large volumes (up to 2001), of water in a single step. Values obtained varied from 18.4 to values higher than 55 MPNCU 1⁻¹in the Besôs river and 0.44-44 MPNCU 1⁻¹in the Llobregat river. Viruses were isolated from approx. 70% of sea-water samples, the values ranging from 0 to 1.60 MPNCU 1⁻¹. Findings are discussed in terms of correlation between virus pollution and bacterial indicators of fecal contamination. Also, the suitability of glass powder adsorption to concentrate viruses in sea-water is discussed.