Spatial variability of soil properties under Pinus canariensis canopy in two contrasting soil textures

The aim of this study was to investigate how the biomass of extramatrical mycorrhizal mycelia (EMM) is influenced by the addition of different phosphorus (P), potassium (K) and PK fertilizers in peatland forests with variable P and K availability. Four fertilizers were used: apatite, biotite, Rauta-PK? (apatite and ferrosulphate) and a test fertilizer (50% apatite and 50% recycled iron phosphate). Forest plots with four different types of nutrient balance (deficient P and deficient K, deficient P and sufficient K, sufficient P and deficient K, and sufficient P and sufficient K) were studied. The effects on EMM biomass and ectomycorrhizal (EM) biomass in roots were estimated by ergosterol and phospholipid fatty acid (PLFA) analysis using in-growth mesh bags. Nutrients and rare-earth elements in EM roots surrounding the mesh bags were quantified and used as indicators of nutrient transport by the EMM in the mesh bags. The biomass of EMM was enhanced by P, K and PK deficiency of the trees, and EM fungal biomass in the roots was increased by P deficiency. The test fertilizer enhanced EMM biomass in all the plots studied, whereas the other fertilizers did not have any significant effect. No significant interactions between the P and K availability of host and mycelial fertilizer response could be detected. Deficiency of P or K or both in needles did not affect the concentrations of rare-earth elements in the tree roots. Earlier results from laboratory experiments have shown reduced carbon allocation to EM fungi under K deficiency, but this was not the case in these mature forests. Instead, we observed increased EMM biomass in response to both P and K deficiency.     

Spatial risk assessment of livestock exposure to pumas in Patagonia, Argentina

Livestock predation and associated human-carnivore conflicts are increasing worldwide and require the development of methods and concepts for risk assessment and conflict management. Here we use knowledge on habitat preference and distribution of pumas and provide a first assessment of the spatial risk of livestock to puma depredation in Patagonian ranches, Argentina. In an initial step, we developed a rule-based habitat model in a Geographic Information System (GIS) to predict the distribution of puma habitat at a regional scale in Patagonia. We then used empirically derived puma occurrence records from Patagonian ranches 1) to test our regional habitat predictions, and 2) to evaluate if paddock characteristics (vegetation cover, topography, and distance to roads) contribute to explain puma occurrences within ranches. Finally, we simulated three livestock management scenarios differing in their spatial and seasonal allocation of livestock to paddocks, and compared the likelihood of livestock exposure to pumas among scenarios. At a regional scale, 22% of the study region was predicted to be suitable for puma home ranges. The greatest uncertainty in these predictions resulted from assumptions on woody vegetation cover requirements at the home range scale. Within ranches, puma occurrences were positively associated with paddock topography, woody vegetation cover on paddocks, and proximity to predicted regional puma habitat. Comparing the risk of predation by puma among simulated livestock management scenarios implied that rotating livestock during seasons may help to reduce the likelihood of livestock exposure to pumas. Our results show the usefulness of rule-based habitat models for describing broad-scale carnivore distributions and for aiding risk assessments to mitigate conflicts between predators and human activities.     

Time efficient detection of protein–ligand interactions with the polarization optimized PO-WaterLOGSY NMR experiment

The identification of compounds that bind to a protein of interest is of central importance in contemporary drug research. For screening of compound libraries, NMR techniques are widely used, in particular the Water-Ligand Observed via Gradient SpectroscopY (WaterLOGSY) experiment. Here we present an optimized experiment, the polarization optimized WaterLOGSY (PO-WaterLOGSY). Based on a water flip-back strategy in conjunction with model calculations and numerical simulations, the PO-WaterLOGSY is optimized for water polarization recovery. Compared to a standard setup with the conventional WaterLOGSY, time consuming relaxation delays have been considerably shortened and can even be omitted through this approach. Furthermore, the robustness of the pulse sequence in an industrial setup was increased by the use of hard pulse trains for selective water excitation and water suppression. The PO-WaterLOGSY thus yields increased time efficiency by factor of 3–5 when compared with previously published schemes. These time savings have a substantial impact in drug discovery, since significantly larger compound libraries can be tested in screening campaigns. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Endothelium-dependent vasorelaxation in rat thoracic aorta by Mansoa hirsuta D.C.

The vasodilator effect of the ethanolic extract of Mansoa hirsuta leaves (EEF) was assayed in rat aortic rings. EEF produced a concentration-dependent vasodilatation (pIC₅₀=5.1±0.2), which was absent in endothelium-denuded vessels. The vasodilator effect of EEF was similar to a standardized ethanolic extract of Hancornia speciosa Gomes (pIC₅₀=5.1±0.1). The endothelium-dependent vasodilatation induced by EEF was abolished by L-NAME (100 μM), a nitric oxide (NO) synthase inhibitor, but not by indomethacin (10 μM; pIC₅₀=4.9±0.2), a cyclooxygenase inhibitor. The concentration–response curve of EEF was not modified by the addition of superoxide dismutase (SOD; 300 U/ml). In addition, EEF (50 μg/ml) displaced the 3-morpholino-sidnonimine (SIN-1; p<0.05) concentration–effect curve to the left, as well as SOD (300 U/ml). These findings lead us to conclude that EEF induces a NO- and endothelium-dependent vasodilatation in rat aortic preparations, and that this effect is, at least in some extent, due to an increase in the NO bioavailability as consequence of its antioxidant activity. The HPLC-DAD profile recorded for EEF indicates the presence of four major peaks with close retention times, exhibiting similar UV spectra with wavelength maxima compatible with heterogeneous proanthocyanidins.     

Grape-seed procyanidins prevent low-grade inflammation by modulating cytokine expression in rats fed a high-fat diet

ObjectiveThe main objective of this study was to evaluate the effect of procyanidin intake on the level of inflammatory mediators in rats fed a hyperlipidic diet, which are a model of low-grade inflammation as they show an altered cytokine production.DesignMale Zucker Fa/fa rats were randomly grouped to receive a low-fat (LF) diet, a high-fat (HF) diet or a high-fat diet supplemented with procyanidins from grape seed (HFPE) (345 mg/kg feed) for 19 weeks and were then euthanized. We determined biochemical parameters, C-reactive protein (CRP) and IL-6 levels in plasma. Adipose tissue depots and body weight were also determined. We assessed CRP, IL-6, TNF-α and adiponectin gene expression in liver and white adipose tissue (WAT).ResultsAs expected, rats fed the HF diet show an enhanced production of CRP. Our results demonstrate that the HFPE diet decreases rat plasma CRP levels but not IL-6 levels. The decrease in plasma CRP in HFPE rats is related to a down-regulation of CRP mRNA expression in the liver and mesenteric WAT. We have also shown a decrease in the expression of the proinflammatory cytokines TNF-α and IL-6 in the mesenteric WAT. In contrast, adiponectin mRNA is increased in this tissue due to the procyanidin treatment.As previously reported, CRP plasma levels correlate positively with its expression in the mesenteric WAT, suggesting that procyanidin extract (PE) modulates CRP at the synthesis level. CRP plasma levels also correlate positively with body weight. As expected, body weight is associated with the adiposity index. Also, TNF-α expression and IL-6 expression have a strong positive correlation. In contrast, the expression of the anti-inflammatory cytokine adiponectin correlates negatively with the expression of TNF-α and IL-6 in the mesenteric WAT.ConclusionThese results suggest a beneficial effect of PE on low-grade inflammatory diseases, which may be associated with the inhibition of the proinflammatory molecules CRP, IL-6 and TNF-α and the enhanced production of the anti-inflammatory cytokine adiponectin. These findings provide a strong impetus to explore the effects of dietary polyphenols in reducing obesity-related adipokine dysregulation to manage cardiovascular and metabolic risk factors.     

Influence of fatty acid composition of raw materials on biodiesel properties

The aim of this work was the study of the influence of the raw material composition on biodiesel quality, using a transesterification reaction. Thus, ten refined vegetable oils were transesterificated using potassium methoxide as catalyst and standard reaction conditions (reaction time, 1h; weight of catalyst, 1 wt.% of initial oil weight; molar ratio methanol/oil, 6/1; reaction temperature, 60 degrees C). Biodiesel quality was tested according to the standard [UNE-EN 14214, 2003. Automotive fuels. Fatty acid methyl esters (FAME) for diesel engines. Requirements and test methods]. Some critical parameters like oxidation stability, cetane number, iodine value and cold filter plugging point were correlated with the methyl ester composition of each biodiesel, according to two parameters: degree of unsaturation and long chain saturated factor. Finally, a triangular graph based on the composition in monounsaturated, polyunsaturated and saturated methyl esters was built in order to predict the critical parameters of European standard for whatever biodiesel, known its composition.     

KCNJ11 knockout morula re-engineered by stem cell diploid aggregation

KCNJ11-encoded Kir6.2 assembles with ATP-binding cassette sulphonylurea receptors to generate ATP-sensitive K+ (KATP) channel complexes. Expressed in tissues with dynamic metabolic flux, these evolutionarily conserved yet structurally and functionally unique heteromultimers serve as high-fidelity rheostats that adjust membrane potential-dependent cell functions to match energetic demand. Genetic defects in channel subunits disrupt the cellular homeostatic response to environmental stress, compromising organ tolerance in the adult. As maladaptation characterizes malignant KATP channelopathies, establishment of platforms to examine progression of KATP channel-dependent adaptive behaviour is warranted. Chimeras provide a powerful tool to assay the contribution of genetic variance to stress intolerance during prenatal or post-natal development. Here, KCNJ11 KATP channel gene knockout<-->wild-type chimeras were engineered through diploid aggregation. Integration of wild-type embryonic stem cells into zona pellucida-denuded morula derived from knockout embryos achieved varying degrees of incorporation of stress-tolerant tissue within the KATP channel-deficient background. Despite the stress-vulnerable phenotype of the knockout, ex vivo derived mosaic blastocysts tolerated intrauterine transfer and implantation, followed by full-term embryonic development in pseudopregnant surrogates to produce live chimeric offspring. The development of adult chimerism from the knockout<-->wild-type mosaic embryo offers thereby a new paradigm to probe the ecogenetic control of the KATP channel-dependent stress response.     

No sex in fungus-farming ants or their crops

Asexual reproduction imposes evolutionary handicaps on asexual species, rendering them prone to extinction, because asexual reproduction generates novel genotypes and purges deleterious mutations at lower rates than sexual reproduction. Here, we report the first case of complete asexuality in ants, the fungus-growing ant Mycocepurus smithii, where queens reproduce asexually but workers are sterile, which is doubly enigmatic because the clonal colonies of M. smithii also depend on clonal fungi for food. Degenerate female mating anatomy, extensive field and laboratory surveys, and DNA fingerprinting implicate complete asexuality in this widespread ant species. Maternally inherited bacteria (e.g. Wolbachia, Cardinium) and the fungal cultivars can be ruled out as agents inducing asexuality. M. smithii societies of clonal females provide a unique system to test theories of parent–offspring conflict and reproductive policing in social insects. Asexuality of both ant farmer and fungal crop challenges traditional views proposing that sexual farmer ants outpace coevolving sexual crop pathogens, and thus compensate for vulnerabilities of their asexual crops. Either the double asexuality of both farmer and crop may permit the host to fully exploit advantages of asexuality for unknown reasons or frequent switching between crops (symbiont reassociation) generates novel ant–fungus combinations, which may compensate for any evolutionary handicaps of asexuality in M. smithii.     

The O-Mannosyltransferase PMT4 Is Essential for Normal Appressorium Formation and Penetration in Ustilago maydis

In Saccharomyces cerevisiae, the PMT, KRE2/MNT1, and MNN1 mannosyltransferase protein families catalyze the steps of the O-mannosylation pathway, sequentially adding mannoses to target proteins. We have identified members of all three families and analyzed their roles in pathogenesis of the maize smut fungus Ustilago maydis. Furthermore, we have shown that PMT4, one of the three PMT family members in U. maydis, is essential for tumor formation in Zea mays. Significantly, PMT4 seems to be required only for pathogenesis and is dispensable for other aspects of the U. maydis life cycle. We subsequently show that the deletion of pmt4 results in a strong reduction in the frequency of appressorium formation, with the few appressoria that do form lacking the capacity to penetrate the plant cuticle. Our findings suggest that the O-mannosylation pathway plays a key role in the posttranslational modification of proteins involved in the pathogenic development of U. maydis. The fact that PMT homologs are not found in plants may open new avenues for the development of fungal control strategies. Moreover, the discovery of a highly specific requirement for a single O-mannosyltransferase should aid in the identification of the proteins directly involved in fungal plant penetration, thus leading to a better understanding of plant–fungi interactions.