Anionic Phospholipid Interactions of the Prion Protein N Terminus Are Minimally Perturbing and Not Driven Solely by the Octapeptide Repeat Domain

Although the N terminus of the prion protein (PrP^C) has been shown to directly associate with lipid membranes, the precise determinants, biophysical basis, and functional implications of such binding, particularly in relation to endogenously occurring fragments, are unresolved. To better understand these issues, we studied a range of synthetic peptides: specifically those equating to the N1 (residues 23–110) and N2 (23–89) fragments derived from constitutive processing of PrP^C and including those representing arbitrarily defined component domains of the N terminus of mouse prion protein. Utilizing more physiologically relevant large unilamellar vesicles, fluorescence studies at synaptosomal pH (7.4) showed absent binding of all peptides to lipids containing the zwitterionic headgroup phosphatidylcholine and mixtures containing the anionic headgroups phosphatidylglycerol or phosphatidylserine. At pH 5, typical of early endosomes, quartz crystal microbalance with dissipation showed the highest affinity binding occurred with N1 and N2, selective for anionic lipid species. Of particular note, the absence of binding by individual peptides representing component domains underscored the importance of the combination of the octapeptide repeat and the N-terminal polybasic regions for effective membrane interaction. In addition, using quartz crystal microbalance with dissipation and solid-state NMR, we characterized for the first time that both N1 and N2 deeply insert into the lipid bilayer with minimal disruption. Potential functional implications related to cellular stress responses are discussed.     

Volatiles modulate the development of plant pathogenic rust fungi

Rust fungi are obligate biotrophic pathogens that differentiate a series of specialized cells to establish infection. One of these cells, the haustorium, which serves to absorb nutrients from living host cells, normally develops only in planta. Here, we show that the rust fungus Uromyces fabae (Pers.) Schroet. stimulates volatile emission of its host, broad bean (Vicia faba L.). Volatiles were identified and shown to be perceived by the fungus in in vitro assays that excluded the host. Three of them, nonanal, decanal, and hexenyl acetate promoted the development of haustoria on artificial membranes. In contrast, the terpenoid farnesyl acetate suppressed this differentiation. In assays using whole plants, farnesyl acetate reduced rust disease not only on broad bean but also on several cereals and legumes including soybean. This natural substance was effective against all rusts tested when directly applied to the host. This demonstrated that farnesyl acetate may serve as a powerful novel tool to combat rust fungi including Phakopsora pachyrhizi that currently threatens the production of soybeans world-wide.     

Profiling of structurally labile oxylipins in plants by in situ derivatization with pentafluorobenzyl hydroxylamine

A GC-MS-based method for the simultaneous quantification of common oxylipins along with labile and highly reactive compounds based on in situ derivatization with pentafluorobenzyl hydroxylamine to the corresponding O-2,3,4,5,6-pentafluorobenzyl oximes (PFB oximes) is presented. The approach covers oxo derivatives such as jasmonic acid (JA), 12-oxophytodienoic acid (OPDA), certain phytoprostanes, unsaturated oxo-acids, oxo-hydroxy acids, and aldehyde fragments from the polar head of fatty acids. In the positive electron impact-MS mode, the PFB oximes display characteristic fragment ions that greatly facilitate the identification of oxylipins in complex matrices. In addition, the fluorinated derivatives allow a highly selective and low-background analysis by negative chemical ionization. Besides showing the general value of the method for the identification of a broad range of oxylipins (18 examples), we also demonstrate sensitivity, linearity, and reproducibility for the quantification of JA, OPDA, 11-oxo-9-undecenoic acid, and 13-oxo-9,11-tridecadienoic acid. The efficiency of the method is demonstrated by differential profiling of these four oxylipins in lima bean leaves after mechanical wounding and feeding by the herbivore Spodoptera littoralis. Caterpillar feeding induced several oxylipins, whereas after wounding only the level of JA increased. The rapid in situ derivatization prevents the isomerization of cis-JA to trans-JA. The resting level of JA in lima beans showed an isomer ratio of 80:20 for trans/cis-JA. After wounding, de novo synthesis of JA alters the ratio to 20:80 in favor of the cis isomer.     

Training-specific changes in cardiac structure and function: a prospective and longitudinal assessment of competitive athletes.

This prospective, longitudinal study examined the effects of participation in team-based exercise training on cardiac structure and function. Competitive endurance athletes (EA, n = 40) and strength athletes (SA, n = 24) were studied with echocardiography at baseline and after 90 days of team training. Left ventricular (LV) mass increased by 11% in EA (116 +/- 18 vs. 130 +/- 19 g/m(2); P < 0.001) and by 12% in SA (115 +/- 14 vs. 132 +/- 11 g/m(2); P < 0.001; P value for the compared Delta = NS). EA experienced LV dilation (end-diastolic volume: 66.6 +/- 10.0 vs. 74.7 +/- 9.8 ml/m(2), Delta = 8.0 +/- 4.2 ml/m(2); P < 0.001), enhanced diastolic function (lateral E': 10.9 +/- 0.8 vs. 12.4 +/- 0.9 cm/s, P < 0.001), and biatrial enlargement, while SA experience LV hypertrophy (posterior wall: 4.5 +/- 0.5 vs. 5.2 +/- 0.5 mm/m(2), P < 0.001) and diminished diastolic function (E' basal lateral LV: 11.6 +/- 1.3 vs. 10.2 +/- 1.4 cm/s, P < 0.001). Further, EA experienced right ventricular (RV) dilation (end-diastolic area: 1,460 +/- 220 vs. 1,650 +/- 200 mm/m(2), P < 0.001) coupled with enhanced systolic and diastolic function (E' basal RV: 10.3 +/- 1.5 vs. 11.4 +/- 1.7 cm/s, P < 0.001), while SA had no change in RV parameters. We conclude that participation in 90 days of competitive athletics produces significant training-specific changes in cardiac structure and function. EA develop biventricular dilation with enhanced diastolic function, while SA develop isolated, concentric left ventricular hypertrophy with diminished diastolic relaxation.     

Chemical defense of brown algae (Dictyopteris spp.) against the herbivorous amphipod Ampithoe longimana

Terpenoids, polyphenols, and C₁₁ metabolites are broadly distributed among brown seaweeds. Terpenoids and polyphenols have often been investigated as chemical defenses against herbivores, while there are only few investigations of the fatty-acid-derived C₁₁ hydrocarbons and C₁₁ sulfur compounds as potential defenses. We investigated effects of C₁₁ sulfur metabolites from the cosmopolitan brown alga Dictyopteris membranacea on feeding and fitness of the herbivorous amphipod Ampithoe longimana. In choice tests between freshly collected thalli of D. hoytii (which lacks C₁₁ sulfur compounds) and D. membranacea (which contains C₁₁ sulfur compounds) amphipods consumed about 4 times more of the species lacking the C₁₁ sulfur compounds. The same feeding preference was observed when these plants were finely ground and embedded in an agar matrix to destroy morphological differences. When a diet made from field-collected thalli of D. membranacea containing C₁₁ sulfur compounds was tested against a diet made from a laboratory culture of D. membranacea that had lost the ability to produce C₁₁ sulfur compounds, the same magnitude of preference was observed for the population lacking the sulfur compounds. In addition to the C₁₁ sulfur compounds, a water-soluble C9-oxo acid that appears to be a by-product in the biosynthesis of the C₁₁ metabolites also suppressed amphipod feeding to a comparable extent. Both classes of compound may contribute to the effective chemical protection of D. membranacea. When juvenile amphipods were reared for 28 days on artificial diets containing the above compounds, their survivorship (⢪%) closely resembled that of a starved treatment, but differed dramatically from a control treatment (60%) consisting of the same food, but without the metabolites. Most other classes of brown algal secondary metabolites are defensive against a broad spectrum of larger herbivores, but relatively ineffective against the amphipod studied here. In contrast, the fatty-acid-derived sulfur compounds and the C9-oxo acid strongly deter Ampithoe-like mesograzers but appear less effective against other herbivores, suggesting that these metabolites could be ecologically important in defending zygotes and germlings against these small consumers.     

An Experimental Test of Predator Detection Rates Using Groups of Free-living Emus

Abstract Improved predator detection is often stated to be one of the principal benefits of social foraging. However, actual field evidence supporting this assumption is scarce. This may be the result of the fact that most observations are conducted on social animals acting in the absence of an acute predation threat, yet the benefits of grouping come to the fore in that brief moment when an individual's life is at risk. As predation attempts are typically rare in nature, experimental manipulations are necessary to further explore the costs and benefits of social foraging. This study utilizes simple predator simulations (by humans) to experimentally test the predator‐detecting abilities and escape strategies of groups of free‐living emus Dromaius novaehollandiae. Emus in larger groups spent less time in vigilance and more time foraging. Nonetheless, the combined vigilance of group members ensured that emus detected the ‘simulated predator’ sooner as group size increased. After detecting the ‘predator’, larger groups waited longer until opting to flee, and then spent less time and energy doing so. Thus, the results of this study provide experimental evidence that emus benefit from grouping in terms of both the ‘many‐eyes effect’ and the ‘dilution effect’.     

Mechanism of ATP hydrolysis by sarcoplasmic reticulum and the role of phospholipids.

Exchange of sarcoplasmic reticulum phospholipids with dipalmitoyllecithin inhibits the (Mg2+ + Ca2+)-activated ATPase activity below 40 degrees by inhibition of the decomposition of phosphoprotein intermediate. The rate of phosphoprotein formation and the steady state concentration of phosphoprotein measured by rapid kinetic techniques are affected to a lesser extent. The inhibitory effect of dipalmitoyllecithin on ATPase activity is probably related to the viscosity of the hydrocarbon region of the membrane which inhibits the conformational change leading to calcium translocation and the eventual cleavage of phosphoprotein.     

Enzymes of ammonia assimilation in legume nodules: A comparison between ureide- and amide-transporting plants

Levels of amide and ureide biogenic enzymes were compared in the plant cytosol fractions of root nodules from soybean ( Glycine max L. Merr., cv. Williams), pintobean ( Phaseolus vulgaris L. cv. Pinto) and Lupin ( Lupinus angustifolius L. cv. Frost). Enzymes of purine oxidation were found to be present in significant quantities only in ureide-transporting pintobean and soybean nodules. The levels of these enzymes were low in lupin, but this amide-exporter had significantly higher levels of asparagine synthetase. Enzymes of de novo purine biosynthesis and glycine biosynthesis were present at higher levels in pintobean and soybean, consistent with a role for de novo purine biosynthesis in ureide biogenesis. The low levels of these enzymes in lupin are consistent with a role in general purine and amino acid metabolism in these nodules, not directly related to the synthesis of transport compounds for fixed atmospheric nitrogen. Amino acid concentrations in soybean, pintobean and lupin nodules reflected the metabolic differences between amide and ureide plants. The comparative data presented are consistent with a pathway of ureide biogenesis using glutamine, glutamate and aspartate synthesized via reactions catalyzed by glutamine synthetase, glutamate synthase and aspartate aminotransferase in the de novo synthesis of purines followed by oxidation of these purines to produce the ureides allantoin and allantoic acid.