Determination of boldine in plasma by high-performance liquid chromatography

A sensitive method for the determination of boldine in blood plasma is described. The procedure involves a direct pH-buffered chloroform extraction of boldine from blood plasma, followed by its assay under isocratic conditions by HPLC with UV detection. The extraction recovery is excellent, and sensitivity and precision of the method are very high, when applied to plasma samples containing pharmacologically relevant concentrations of boldine.     

The role of parental and hybrid species in multiple introgression events: evidence of homoploid hybrid speciation in Centaurea (Cardueae,Asteraceae)

Molecular approaches have greatly increased the number of confirmed homoploid hybrids, which suggests that the frequency of this phenomenon was underestimated in the past because it was much more difficult to detect than allopolyploidy. Centaurea is a suitable model group for studying homoploid speciation, as hybridization events have been commonly reported for this genus. Based on this, here we study Centaurea × forsythiana, a naturally occurring homoploid hybrid between two Sardinian endemics, C. horrida and C. filiformis, using a molecular approach involving nuclear and plastid markers, to understand the underlying population dynamics between homoploid hybrids and their parents. Our results confirm that C. × forsythiana is a hybrid between the above‐mentioned species and define the roles of the parents. Plastid markers point towards C. horrida as the maternal progenitor, and nuclear markers reveal that the other parental species, C. filiformis, is itself an old, stabilized homoploid hybrid related to the C. paniculata complex from the Italian mainland. Homoploid hybrid speciation is discussed and C. × forsythiana and C. filiformis are compared with other similar examples. The study confirms the importance of introgression between parental species mediated by hybrids and its potential implications in conservation. Furthermore, it shows how hybridization studies become even more complex when the parents are themselves of probable hybrid origin. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 453–467.     

Early steps of adventitious rooting: morphology,hormonal profiling and carbohydrate turnover in carnation stem cuttings

The rooting of stem cuttings is a common vegetative propagation practice in many ornamental species. A detailed analysis of the morphological changes occurring in the basal region of cultivated carnation cuttings during the early stages of adventitious rooting was carried out and the physiological modifications induced by exogenous auxin application were studied. To this end, the endogenous concentrations of five major classes of plant hormones [auxin, cytokinin (CK), abscisic acid, salicylic acid (SA) and jasmonic acid] and the ethylene precursor 1‐aminocyclopropane‐1‐carboxylic acid were analyzed at the base of stem cuttings and at different stages of adventitious root formation. We found that the stimulus triggering the initiation of adventitious root formation occurred during the first hours after their excision from the donor plant, due to the breakdown of the vascular continuum that induces auxin accumulation near the wounding. Although this stimulus was independent of exogenously applied auxin, it was observed that the auxin treatment accelerated cell division in the cambium and increased the sucrolytic activities at the base of the stem, both of which contributed to the establishment of the new root primordia at the stem base. Further, several genes involved in auxin transport were upregulated in the stem base either with or without auxin application, while endogenous CK and SA concentrations were specially affected by exogenous auxin application. Taken together our results indicate significant crosstalk between auxin levels, stress hormone homeostasis and sugar availability in the base of the stem cuttings in carnation during the initial steps of adventitious rooting.     

Mapping impaired olive tree development using electromagnetic induction surveys

Background and aims

Future success of olive cropping in the Mediterranean depends critically on improving yield, reducing production costs, and preventing infestation by soil-borne pathogens. In order to put forward adequate soil management practices accurate knowledge of the spatial distribution of soil properties is required. The aims of this study were to delimit areas with constrained tree development in an olive orchard using electromagnetic induction (EMI), and to identify the causal relationships between apparent electrical conductivity (ECa) and soil properties.

Methods

The experimental field was exhaustively sampled for different soil properties and ECa was measured in 2011 and 2012 under dry and wetter soil conditions, respectively.

Results

The spatial ECa distribution matched the observed canopy coverage pattern well. Three zones were delimited according to ECa values from 0 to 27.5, from 27.5 to 57.5, and greater than 57.5 mS m^?1. All ECa signals, regardless of soil-water status, exhibited a common dominant ECa pattern. The area with the lowest ECa values (0–27.5 mS m^?1) showed optimal tree growth (45 % canopy coverage) and presented significantly lower average clay contents than the other two areas. Intermediate ECa values (27.5–57.5 mS m^?1) identified accurately the area with deficient tree development and tree die-off (12 % canopy coverage), and corresponded with an area along the drainage pathway where profile-averaged soil-water, clay, stone and organic matter content were highest.

Conclusions

EMI surveys detected subtle differences in soil properties and provided useful information to delimit areas with constrained tree development. The approach can be used as a screening technique before installing tree plantations.     

Ecological responses of aquatic macrophytes and benthic macroinvertebrates to dams in the Henares River Basin (Central Spain)

The ecological responses of aquatic macrophytes and benthic macroinvertebrates to deep-release dams in three impounded rivers of the Henares River Basin (Central Spain) were studied, specially focusing on the effects of nutrient enrichment caused by deep releases on these two freshwater communities. Three sampling sites, one upstream and two downstream from the reservoir, were established in each impounded river. Sampling surveys to collect submersed macrophytes and benthic macroinvertebrates at each sampling site were carried out in spring–summer of 2009 and 2011. Water temperature tended to decrease downstream from dams, whereas nitrate and phosphate concentrations tended to increase. These abiotic changes, particularly the downstream nutrient enrichment, apparently affected the macrophyte and macroinvertebrate communities. In the case of submersed macrophytes, total coverage and taxa richness increased downstream from dams. In the case of benthic macroinvertebrates, total density and total biomass also increased downstream, but taxa richness tended to decrease. Scrapers appeared to be the macroinvertebrate feeding group most favored downstream from dams as a probable consequence of the positive effect of nutrient enrichment on periphyton and perilithon abundance. Nutrients would ultimately come from water runoff over agricultural lands and over semi-natural forests and pastures, being subsequently accumulated in the hypolimnion of reservoirs.     

Conformational Recognition of an Intrinsically Disordered Protein

There is a growing interest in understanding the properties of intrinsically disordered proteins (IDPs); however, the characterization of these states remains an open challenge. IDPs appear to have functional roles that diverge from those of folded proteins and revolve around their ability to act as hubs for protein-protein interactions. To gain a better understanding of the modes of binding of IDPs, we combined statistical mechanics, calorimetry, and NMR spectroscopy to investigate the recognition and binding of a fragment from the disordered protein Gab2 by the growth factor receptor-bound protein 2 (Grb2), a key interaction for normal cell signaling and cancer development. Structural ensemble refinement by NMR chemical shifts, thermodynamics measurements, and analysis of point mutations indicated that the population of preexisting bound conformations in the free-state ensemble of Gab2 is an essential determinant for recognition and binding by Grb2. A key role was found for transient polyproline II (PPII) structures and extended conformations. Our findings are likely to have very general implications for the biological behavior of IDPs in light of the evidence that a large fraction of these proteins possess a specific propensity to form PPII and to adopt conformations that are more extended than the typical random-coil states.