Analytical discrimination of poisonous and nonpoisonous chemotypes of giant fennel (Ferula communis L.) through their biologically active and volatile fractions

Giant fennel (Ferula communis L.) from Sardinia is characterized by two chemotypes with different biological activities. One chemotype is poisonous, due to prenylcoumarins, and responsible for ferulosis, which mainly affects sheep and goats, cattle, and horses; the other chemotype is nonpoisonous and contains daucane esters. The two chemotypes cannot be distinguished botanically. High-performance liquid chromatography-diode array-ultraviolet detection-mass spectrometry (HPLC-DAD-UV-MS) analysis of the composition of the fractions containing the biologically active metabolites and of the volatile fractions, by gas chromatography-mass spectrometry (GC-MS), of both essential oil and headspace sampled by headspace solid-phase microextraction (HS-SPME) are here shown to be effective in discriminating the poisonous and nonpoisonous chemotypes. HS-SPME with CAR/PDMS/DVB in combination with GC-MS has also been found to be a successful, fully automated one-step method for rapid and unequivocal discrimination of the two chemotypes, using aristolene and allohedycaryol as markers of the poisonous and nonpoisonous chemotypes, respectively.     

Topography and crop management are key factors for the development of american leaf spot epidemics on coffee in costa rica

ABSTRACT We monitored the development of American leaf spot of coffee, a disease caused by the gemmiferous fungus Mycena citricolor, in 57 plots in Costa Rica for 1 or 2 years in order to gain a clearer understanding of conditions conducive to the disease and improve its control. During the investigation, characteristics of the coffee trees, crop management, and the environment were recorded. For the analyses, we used partial least-squares regression via the spline functions (PLSS), which is a nonlinear extension to partial least-squares regression (PLS). The fungus developed well in areas located between approximately 1,100 and 1,550 m above sea level. Slopes were conducive to its development, but eastern-facing slopes were less affected than the others, probably because they were more exposed to sunlight, especially in the rainy season. The distance between planting rows, the shade percentage, coffee tree height, the type of shade, and the pruning system explained disease intensity due to their effects on coffee tree shading and, possibly, on the humidity conditions in the plot. Forest trees and fruit trees intercropped with coffee provided particularly propitious conditions. Apparently, fertilization was unfavorable for the disease, probably due to dilution phenomena associated with faster coffee tree growth. Finally, series of wet spells interspersed with dry spells, which were frequent in the middle of the rainy season, were critical for the disease, probably because they affected the production and release of gemmae and their viability. These results could be used to draw up a map of epidemic risks taking topographical factors into account. To reduce those risks and improve chemical control, our results suggested that farmers should space planting rows further apart, maintain light shading in the plantation, and prune their coffee trees.     

Fast and local assessment of stilbene content in grapevine leaf by in vivo fluorometry

Stilbenes are grapevine phytoalexins. These highly fluorescent molecules are generally analyzed by HPLC. This technique allows accurate assay of different stilbenes, but it is destructive, time-consuming, and neglects their spatial distribution. This is why we have tested a new method based on in vivo fluorescence using commercial spectrofluorometers that allowed fast and local assessment of stilbene content in grapevine leaves. Stilbene synthesis in grapevine Vitis vinifera var. Muscat Ottonel leaves was induced by Plasmopara viticola inoculation or UV-C irradiation. Fluorescence was measured both from the abaxial and adaxial sides of leaves, then stilbene content was analyzed by HPLC. It varied from 0 in control leaves to 15 mg g-1 dry weight in UV-treated leaves. Highly significant regressions were found between HPLC stilbene content and the corresponding leaf UV-induced blue fluorescence. Thus, in vivo fluorescence is a good tool for a rapid study of stilbenes synthesis in grapevine leaves that can potentially be extended to other fluorescent molecules.     

Autoantibodies against the chromosomal passenger protein INCENP found in a patient with Graham Little-Piccardi-Lassueur syndrome

Background

Graham Little – Piccardi – Lassueur (GLPL) syndrome is a rare dermatosis characterized by scarring alopecia, loss of pubic and axillary hair, and progressive development of variously located follicular papules. We report a first case ever of an autoimmune response in a patient suffering from GLPL syndrome.

Methods

Immunofluorescence and immunoblot analysis were used in a variety of cell cultures including human, monkey, hamster, mouse and bovine cells to analyze the presence of autoantibodies in a GLPL patient.

Results

The autoimmune serum showed a pattern of centromere and spindle microtubule staining resembling that of the chromosomal passenger protein complex. By using a complex of proteins expressed in baculovirus, immunoblot analysis demonstrated that the INCENP protein is a major autoantigen in this patient with GLPL syndrome.

Conclusion

An autoimmune response in GLPL syndrome is reported against the INCENP centromere protein. The occasional development of autoimmunity in GLPL patients could serve as a test in continuing efforts to detect this disease and for a more directed therapy based on the autoantigen response.

     

Composition and cellular localization of tannins in grape seeds during maturation

Cell walls were isolated from seeds of grape berries (Vitis vinifera L.), and proanthocyanidin composition was determined over the course of ripening for different levels of vine water status. During the ripening period the tannins from the cell walls were always more polymerized than those from the inner part of the cell. At maturity this difference becomes more significant compared to véraison, due to a significant increase in the mean degree of polymerization of the cell wall tannins. The tannin composition was typical of grape seed tannins and was quite similar in the two cell fractions studied, but the epicatechin gallate proportion was significantly higher in the cell wall fraction. There were no significant effects of water deficit on composition and polymerization of seed tannins.     

Extracts enriched in different polyphenolic families normalize increased cardiac NADPH oxidase expression while having differential effects on insulin resistance, hypertension, and cardiac hypertrophy in high-fructose-fed rats

Insulin resistance and oxidative stress act synergistically in the development of cardiovascular complications. The present study compared the efficacy of three polyphenolic extracts in their capacity to prevent hypertension, cardiac hypertrophy, increased production of reactive oxygen species (ROS) by the aorta or the heart, and increased expression of cardiac NAD(P)H oxidase in a model of insulin resistance. Rats were fed a 60%-enriched fructose food and were treated once a day (gavage) for 6 weeks with 10 mL/kg of water only (F group) or the same amount of solution containing a red grape skin polyphenolic extract enriched in anthocyanins (ANT), a grape seed extract enriched in procyanidins and rich in galloylated procyanidins (PRO), or the commercial preparation Vitaflavan (VIT), rich in catechin oligomers. All treatments were administered at the same dose of 21 mg/kg of polyphenols. Our data indicate that (a) the ANT treatment prevented hypertension, cardiac hypertrophy, and production of ROS, (b) the PRO treatment prevented insulin resistance, hypertriglyceridemia, and overproduction of ROS but had only minor effects on hypertension or hypertrophy, while (c) Vitaflavan prevented hypertension, cardiac hypertrophy, and overproduction of ROS. All polyphenolic treatments prevented the increased expression of the p91phox NADPH oxidase subunit. In summary, our study suggest that (a) the pathogeny of cardiac hypertrophy in the fructose-fed rat disease involves both hypertension and hyperproduction of ROS, (b) polyphenolic extracts enriched in different types of polyphenols possess differential effects on insulin resistance, hypertension, and cardiac hypertrophy, and (c) polyphenols modulate the expression of NAD(P)H oxidase.     

Duricrusts and deep-weathering profiles in southwestern wisconsin

Numerous exposures in southwestern Wisconsin display the profiles of relict deep weathering. Crusts are mainly siliceous, but many are ferruginized to some extent. The occurrence of blocky silcrete, thought to be reported for the first time, recalls the profiles in the Australian belt of transition from silcrust to ferricrust. The humid-tropical kind of pedogenesis necessary to effect this transition may have operated as late as the mid-Miocene. In some localities, the action of grotndwater has caused crusting, mottling, and reduction beneath the dolomite low surface moisture.     

Active surveillance of bat rabies in France: a 5-year study (2004-2009)

Active surveillance of bats in France started in 2004 with an analysis of 18 of the 45 bat species reported in Europe. Rabies antibodies were detected in six indigenous species, mainly in Eptesicus serotinus and Myotis myotis, suggesting previous contact with the EBLV-1 rabies virus. Nineteen of the 177 tested bats were shown serologically positive in seven sites, particularly in central and south-western France. Neither infectious viral particles nor viral genomes were detected in 173 and 308 tested oral swabs, respectively. The presence of neutralising antibodies in female bats (18.6%) was significantly higher than in males (5.6%).     

Near-perfect elastoplasticity in pure nanocrystalline copper

Ductile metals and alloys undergo plastic yielding at room temperature, during which they exhibit work-hardening and the generation of surface instabilities that lead to necking and failure. We show that pure nanocrystalline copper behaves differently, displaying near-perfect elastoplastic behavior characterized by Newtonian flow and the absence of both work-hardening and neck formation. We observed this behavior in tensile tests on fully dense large-scale bulk nanocrystalline samples. The experimental results further our understanding of the unique mechanical properties of nanocrystalline materials and also provide a basis for commercial technologies for the plastic (and superplastic) formation of such materials.     

Recombinant antibodies to the small GTPase Rab6 as conformation sensors

Here we report an approach, based on antibody phage display, to generate molecular conformation sensors. Recombinant antibodies specific to the guanosine triphosphate (GTP)-bound conformation of the small guanosine triphosphatase (GTPase) Rab6, a regulator of membrane traffic, were generated and used to locate Rab6.GTP in fixed cells, and, after green fluorescent protein (GFP) tagging and intracellular expression, to follow Rab6.GTP in vivo. Rab6 was in its GTP-bound conformation on the Golgi apparatus and transport intermediates, and the geometry of transport intermediates was modulated by Rab6 activity. More generally, the same approach could be applied to other molecules that can be locked in a particular conformation in vitro.