Structural effects of the binding of GTP to the wild-type and oncogenic forms of the ras-gene-encoded p21 proteins

Molecular dynamics calculations have been performed to determine the average structures of ras-gene-encoded p21 proteins bound to GTP, i.e., the normal (wild-type) protein and two oncogenic forms of this protein, the Val 12- and Leu 61-p21 proteins. We find that the average structures for all of these proteins exhibit low coordinate fluctuations (which are highest for the normal protein), indicating convergence to specific structures. From previous dynamics calculations of the average structures of these proteins bound to GDP, major regional differences were found among these proteins [Monaco et al. (1995), J. Protein Chem., in press]. We now find that the average structures of the oncogenic proteins are more similar to one another when the proteins are bound to GTP than when they are bound to GDP [Monaco et al. (1995), J. Protein Chem., in press]. However, they still differ in structure at specific amino acid residues rather than in whole regions, in contradistinction to the results found for the p21-GDP complexes. Two exceptions are the regions 25-32, in an alpha-helical region, and 97-110. The two oncogenic (Val 12- and Leu 61-) proteins have similar structures which differ significantly in the region of residues 97-110. This region has recently been identified as being critical in the interaction of p21 with kinase target proteins. The differences in structure between the oncogenic proteins suggest the existence of more than one oncogenic form of the p21 protein that can activate different signaling pathways.     

Selective reduction of GluR2 protein in adult hippocampal CA3 neurons following status epilepticus but prior to cell loss

Kainic acid (KA) induces status epilepticus and delayed neurodegeneration of CA3 hippocampal neurons. Downregulation of glutamate receptor 2 (GluR2) subunit mRNA [the alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) subunit that limits Ca2+ permeability] is thought to a play role in this neurodegeneration, possibly by increased formation of Ca2+ permeable AMPA receptors. The present study examined early hippocampal decreases in GluR2 mRNA and protein following kainate-induced status epilepticus and correlated expression changes with the appearance of dead or dying cells by several histological procedures. At 12 h, in situ hybridization followed by emulsion dipping showed nonuniform decreases in GluR2 mRNA hybridization grains overlying morphologically healthy-appearing CA3 neurons. GluR1 and N-methyl-D-aspartate receptor mRNAs were unchanged. At 12-16 h, when little argyrophilia or cells with some features of apoptosis were detected by silver impregnation or electron microscopy, single immunohistochemistry with GluR2 and GluR2/3 subunit-specific antibodies demonstrated a pattern of decreased GluR2 receptor protein within CA3 neurons that appeared to predict a pattern of damage, similar to the mRNA observations. Double immunolabeling showed that GluR2 immunofluorescence was depleted and that GluR1 immunofluorescence was sustained in clusters of the same CA3 neurons. Quantitation of Western blots showed increased GluR1:GluR2 ratios in CA3 but not in CA1 or dentate gyrus subfields. Findings indicate that the GluR1:GluR2 protein ratio is increased in a population of CA3 neurons prior to significant cell loss. Data are consistent with the "GluR2 hypothesis" that reduced expression of GluR2 subunits will increase formation of AMPA receptors permeable to Ca2+ and predict vulnerability to a particular subset of pyramidal neurons following status epilepticus.     

Dynamics of cyanide binding to ferrous Scapharca inaequivalvis homodimeric hemoglobin

Flash photolysis experiments have been carried out for the first time on a hemoglobin ferrous cyanide adduct with an 8 ns laser pulse. A 95% nonexponential rebinding process occurs within 2 micros after full photolysis in ferrous cyanide dimeric Scapharca inaequivalvis hemoglobin (HbI), indicating that once photolyzed the cyanide anion is not able to escape from the protein matrix and rebinds to the heme iron. The resonance Raman spectrum of the 10 ns photoproduct is identical to that of the fully relaxed deoxy derivative, indicating that in the ferrous cyanide HbI adduct protein relaxation occurs within 10 ns after photolysis. This behavior is at variance with that of the carbonmonoxy HbI derivative in which very little geminate rebinding is observed and the photoproduct relaxes with a lifetime of 1 micros. The fast relaxation of the cyanide HbI photoproduct can be accounted for by the small perturbation of the heme structure induced by cyanide binding to ferrous HbI. This is consistent with a deoxy-like conformation of the HbI ferrous cyanide adduct and implies that the pathway for relaxation involves only minor local rearrangements of the heme moiety. Photolysis experiments carried out on ferrous cyanide horse myoglobin, which can be saturated only partially, show a qualitatively similar behavior in ligand rebinding, indicating that the geminate process of the cyanide anion is a general phenomenon in hemoproteins.     

The attentional demands of encoding and retrieval in younger and older adults: I. Evidence from divided attention costs.

Four studies examined the effects of divided attention in younger and older adults. Attention was divided at encoding or retrieval in free recall (Experiment 1), cued recall (Experiments 2 and 3), and recognition (Experiment 4). Dividing attention at encoding disrupted memory performance equally for the two age groups; by contrast, for both age groups, dividing attention at retrieval had little or no effect on memory performance. Secondary task reaction times (RTs) were slowed to a greater extent for the older adults than for the younger adults, especially at retrieval. Age-related differences in RTs costs at retrieval were largest in free recall, smaller in cued recall, and smallest in recognition. These results provide evidence for an age-related increase in the attentional demands of encoding and retrieval. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Physical and antibacterial properties of edible films formulated with apple skin polyphenols

Fruit and vegetable skins have polyphenolic compounds, terpenes, and phenols with antimicrobial and antioxidant activity. These flavoring plant essential oil components are generally regarded as safe. Edible films made from fruits or vegetables containing apple skin polyphenols have the potential to be used commercially to protect food against contamination by pathogenic bacteria. The main objective of this study was to evaluate physical properties as well as antimicrobial activities against Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica of apple skin polyphenols at 0% to 10% (w/w) concentrations in apple puree film-forming solutions formulated into edible films. Commercial apple skin polyphenol powder had a water activity of 0.44 and high total soluble phenolic compounds and antioxidant capacity (995.3 mg chlorogenic acid/100 g and 14.4 mg Trolox/g, respectively). Antimicrobial activities of edible film containing apple skin polyphenols were determined by the overlay method. Apple edible film with apple skin polyphenols was highly effective against L. monocytogenes. The minimum concentration need to inactive L. monocytogenes was 1.5%. However, apple skin polyphenols did not show any antimicrobial effect against E. coli O157:H7 and S. enterica even at 10% level. The presence of apple skin polyphenols reduced water vapor permeability of films. Apple skin polyphenols increased elongation of films and darkened the color of films. The results of the present study show that apple skin polyphenols can be used to prepare apple-based antimicrobial edible films with good physical properties for food applications by direct contact.     

Thermal patterns in simple models of cylindrical reactors

The propagation of fronts and the emergence of spatiotemporal patterns on a cylindrically shaped thin catalytic reactor is simulated with a homogeneous model of a fixed catalytic bed, with characteristically large Lewis and Peclet numbers, and a first-order Arrhenius kinetics (i.e., thermokinetic model) which may be coupled with slow changes of catalytic activity (i.e., oscillatory kinetics). Planar fronts of the thermokinetic model may undergo symmetry breaking in the transversal direction only at relatively low Lewis number, but for high Le the front remains flat. Patterns due to oscillatory kinetics in reactors of high Le are shown, for the first time, to undergo symmetry breaking in the azimuthal direction when the perimeter is sufficiently large. The generic regular patterns simulated then are rotating multi-wave patterns of constant rotation-speed and oscillatory-‘firing’ ones, and theirs selection is highly sensitive to governing parameters and initial conditions. The results are organized in bifurcation diagrams showing the coexisting two-dimensional solutions with varying perimeter. Increasing convective velocity or reactor radius leads to symmetry breaking of regular patterns and the system may switch to chaos.     

Mass transfer around prolate spheroidal drops in an extensional flow

Mass transfer in the continuous phase around a small eccentricity prolate spheroidal drop in an axisymmetric extensional creeping flow and at large Peclet numbers was investigated theoretically. The results show that, at very short times, the total quantity of solute transferred to or from the drop represents, at O(Ca¹), mass transfer by diffusion only around a sphere. For long times, or at steady‐state, the total quantity of solute transferred is, at O(Ca¹), slightly smaller than that of a spherical drop, and it decreases with an increase of the capillary number or the viscosity ratio. © 2012 Canadian Society for Chemical Engineering     

Arabidopsis thaliana Plants with Different Levels of Aliphatic- and Indolyl-Glucosinolates Affect Host Selection and Performance of Bemisia tabaci

Generalist insects show reduced selectivity when subjected to similar, but not identical, host plant chemical signatures. Here, we produced transgenic Arabidopsis thaliana plants that over-express genes regulating the aliphatic- and indolyl- glucosinolates biosynthetic pathways with either a constitutive (CaMV 35S) or a phloem-specific promoter (AtSUC2). This allowed us to examine how exposure to high levels of aliphatic- or indolyl-glucosinolates in homogenous habitats (leaf cage apparatus containing two wild-type or two transgenic leaves) and heterogeneous habitats (leaf cage apparatus containing one wild-type and one transgenic leaf) affects host selection and performance of Bemsia tabaci, a generalist phloem-feeding insect. Data from homogenous habitats indicated that exposure to A. thaliana plants accumulating high levels of aliphatic- or indolyl-glucosinolates negatively affected the performance of both adult females and nymphs of B. tabaci. Data from heterogeneous habitats indicated that B. tabaci adult females selected for oviposition plants on which their offspring perform better (preference-performance relationship). However, the combinations of wild-type and transgenic plants in heterogeneous habitats increased the period of time until the first choice was made and led to increased movement rate on transgenic plants, and reduced fecundity on wild-type plants. Overall, our findings are consistent with the view that both performance and selectivity of B. tabaci decrease in heterogeneous habitats that contain plants with closely-related chemical signatures.     

Mechanisms of Solution Reactions of Cyclo-Penta[cd]Pyrene Oxide and Acenaphthylene Oxide

Rates and products of the acid-catalyzed and spontaneous reactions of cyclopenta[cd]pyrene oxide (2) and acenaphthalene oxide (3) in 1:3 dioxane-water and water, respectively, have been determined. Both 2 and 3 undergo acid-catalyzed hydrolyses to yield cis diol as the major product, and undergo spontaneous reactions to yield predominantly ketone products. The mechanisms of these reactions are discussed.     

Flow and Pressure Balancing in a Fluid Interface Impactor

Fluid interface impactors, such as virtual and opposing jet impactors, require particular attention to flow and pressure balance in the incoming and outgoing airstreams. The relationship of flow rate to pressure drop has been studied in an opposing jet aerosol classifier of rectangular design. In tests of the sensitivity of classifier operation, changes in pressure differences across the inlets and the outlets were associated with changes in the flow balances of 0.5 and -1.0%/mm w.g., respectively. These flow sensitivities may be used in conjunction with cut size sensitivities to conclude that the appropriate measurements to use for classifier control are the static pressure differences between the two inlets and between the two outlets, rather than the mass flow rates of the four jets. For aerosol size classification tests, a nebulizer-virtual impactor polydisperse aerosol generator was designed to reduce the concentration of small particles and make the generator compatible with dynamic optical particle counting. The aerodynamic cut diameter is Stk ^1/2 ₅₀ = 0.49 ± 0.08 for the standard pressure balance condition. Classification sharpness, using 84 and 16% efficiency sizes, is 1.43 ± 0.03 at the standard condition and can be reduced to 1.11 ± 0.02 at other conditions.