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The efficient conservation of vulnerable ecosystems in the face of global change requires a complete understanding of how plant communities respond to various environmental factors. We aim to demonstrate that a combined use of different approaches, traits, and indices representing each of the taxonomic and functional characteristics of plant communities will give complementary information on the factors driving vegetation assembly patterns. We analyzed variation across an environmental gradient in taxonomic and functional composition, richness, and diversity of the herb-layer of a temperate beech-oak forest that was located in northern Spain. We measured species cover and four functional traits: leaf dry matter content (LDMC), specific leaf area (SLA), leaf size, and plant height. We found that light is the most limiting resource influencing herb-layer vegetation. Taxonomic changes in richness are followed by equivalent functional changes in the diversity of leaf size but by opposite responses in the richness of SLA. Each functional index is related to different environmental factors even within a single trait (particularly for LDMC and leaf size). To conclude, each characteristic of a plant community is influenced by different and even contrasting factors or processes. Combining different approaches, traits, and indices simultaneously will help us understand how plant communities work.

Three Gram-negative, rod-shaped, oxidase-positive, non-spore-forming, non-motile strains (C130915_07T, C150915_16 and C150915_17) were isolated from lymph nodes of Algerian cows. On the basis of 16S rRNA gene and whole genome similarities, the isolates were almost identical and clearly grouped in the genus Pseudochrobactrum . This allocation was confirmed by the analysis of fatty acids (C19:cyclo, C18 : 1, C18 : 0, C16 : 1 and C16 : 0) and of polar lipids (major components: phosphatidylethanolamine, ornithine-lipids, phosphatidylglycerol, cardiolipin and phosphatidylcholine, plus moderate amounts of phosphatidylmonomethylethanolamine, phosphatidyldimethylethanolamine and other aminolipids). Genomic, physiological and biochemical data differentiated these isolates from previously described Pseudochrobactrum species in DNA relatedness, carbon assimilation pattern and growth temperature range. Thus, these organisms represent a novel species of the genus Pseudochrobactrum , for which the name Pseudochrobactrum algeriensis sp. nov. is proposed (type strain C130915_07T=CECT30232T=LMG 32378T).

The efficient conservation of vulnerable ecosystems in the face of global change requires a complete understanding of how plant communities respond to various environmental factors. We aim to demonstrate that a combined use of different approaches, traits, and indices representing each of the taxonomic and functional characteristics of plant communities will give complementary information on the factors driving vegetation assembly patterns. We analyzed variation across an environmental gradient in taxonomic and functional composition, richness, and diversity of the herb-layer of a temperate beech-oak forest that was located in northern Spain. We measured species cover and four functional traits: leaf dry matter content (LDMC), specific leaf area (SLA), leaf size, and plant height. We found that light is the most limiting resource influencing herb-layer vegetation. Taxonomic changes in richness are followed by equivalent functional changes in the diversity of leaf size but by opposite responses in the richness of SLA. Each functional index is related to different environmental factors even within a single trait (particularly for LDMC and leaf size). To conclude, each characteristic of a plant community is influenced by different and even contrasting factors or processes. Combining different approaches, traits, and indices simultaneously will help us understand how plant communities work.

[Background] Climate and land-use changes, which include the application of various types of organic and inorganic fertilisers, have been reducing the species diversity of Mediterranean grasslands and threatening its conservation. Annual plants are one of the most diverse functional groups of species in these grasslands, despite suffering competitive pressure from perennial herbaceous and woody species, and they are essential for ecosystem functioning and stability., [Aims] To quantify how fertilisation modulates the impact of plant-to-plant interactions and climate fluctuations on the dynamics of annuals in Mediterranean grasslands. We hypothesised that the application of sewage sludge would increase competition between functional groups, reducing the abundance of annuals in the long-term, but would buffer the negative impacts of drought on the year-to-year fluctuation of the diversity of annuals., [Methods] In a semi-natural species-rich Mediterranean grassland in northern Spain, we analysed the changes in the taxonomical and functional composition and diversity of annuals over 14 years in response to variations in the abundance of perennial herbaceous and woody species, climate fluctuations and fertilisation with sewage sludge. We quantified separately the patterns of year-to-year fluctuations and long-term trends., [Results] The frequency and diversity of annuals decreased with higher abundance of perennial herbaceous species, drought in June and cold winters. The addition of sewage sludge decreased the abundance of annuals in the long-term, seemed to promote competition between annuals and other functional groups at an interannual scale, and mitigated the negative effects of drought and cold., [Conclusions] Fertilisation influences differently the temporal response of annuals to climate fluctuations and plant-to-plant interactions.

[Background] Climate and land-use changes, which include the application of various types of organic and inorganic fertilisers, have been reducing the species diversity of Mediterranean grasslands and threatening their conservation. Annual plants are one of the most diverse functional groups of species in these grasslands, despite suffering competitive pressure from perennial herbaceous and woody species, and they are essential for ecosystem functioning and stability., [Aims] To quantify how fertilisation modulates the impact of plant-to-plant interactions and climate fluctuations on the dynamics of annuals in Mediterranean grasslands. We hypothesised that the application of sewage sludge would increase competition between functional groups, reducing the abundance of annuals in the long-term, but would buffer the negative impacts of drought on the year-to-year fluctuation of the diversity of annuals., [Methods] In a semi-natural species-rich Mediterranean grassland in northern Spain, we analysed the changes in the taxonomical and functional composition and diversity of annuals over 14 years in response to variations in the abundance of perennial herbaceous and woody species, climate fluctuations, and fertilisation with sewage sludge. We quantified separately the patterns of year-to-year fluctuations and long-term trends., [Results] The frequency and diversity of annuals decreased with a higher abundance of perennial herbaceous species, drought in June, and cold winters. The addition of sewage sludge decreased the abundance of annuals in the long-term, seemed to promote competition between annuals and other functional groups at an interannual scale, and mitigated the negative effects of drought and cold., [Conclusions] Fertilisation influences differently the temporal response of annuals to climate fluctuations and plant-to-plant interactions.

Understanding the mechanisms underlying community stability has become an urgent need in order to protect ecosystems from global change and resulting biodiversity loss. While community stability can be influenced by richness, synchrony in annual fluctuations of species, species stability and functional traits, the relative contributions of these drivers to stability are still unclear. In semi-natural grasslands, land-use changes such as fertilization might affect stability by decreasing richness and influencing year-to-year fluctuations. In addition, they can promote long-term directional trends, shifting community composition and influencing grassland maintenance. Thus, it is important to consider how species and community stability vary year-to-year but also in the long term. Using a 14-year vegetation time series of a species-rich semi-natural Mediterranean grassland, we studied the relative importance of richness, synchrony, species stability and functional traits on community stability. To assess land-use change effects on stability, we applied a fertilization treatment. To distinguish stability patterns produced by year-to-year fluctuations from those caused by long-term trends, we compared the results obtained using a detrending approach from those without detrending. Stability is influenced by richness, synchrony and functional traits. Fertilization decreases species and community stability by promoting long-term trends in species composition, favouring competitive species and decreasing richness. Studying stability at the community and species level, and accounting for the effect of trends is essential to understand stability and its drivers more comprehensively.

Understanding the mechanisms underlying community stability has become an urgent need in order to protect ecosystems from global change and resulting biodiversity loss. While community stability can be influenced by richness, synchrony in annual fluctuations of species, species stability and functional traits, the relative contributions of these drivers to stability are still unclear. In semi-natural grasslands, land-use changes such as fertilization might affect stability by decreasing richness and influencing year-to-year fluctuations. In addition, they can promote long-term directional trends, shifting community composition and influencing grassland maintenance. Thus, it is important to consider how species and community stability vary year-to-year but also in the long term. Using a 14-year vegetation time series of a species-rich semi-natural Mediterranean grassland, we studied the relative importance of richness, synchrony, species stability and functional traits on community stability. To assess land-use change effects on stability, we applied a fertilization treatment. To distinguish stability patterns produced by year-to-year fluctuations from those caused by long-term trends, we compared the results obtained using a detrending approach from those without detrending. Stability is influenced by richness, synchrony and functional traits. Fertilization decreases species and community stability by promoting long-term trends in species composition, favouring competitive species and decreasing richness. Studying stability at the community and species level, and accounting for the effect of trends is essential to understand stability and its drivers more comprehensively., [Methods] Study site and experimental design: In 2003, 12 plots of 15x5 m (hereafter called macro-plots) were established inside an area of 5500 m2. Half of the macro-plots (six) were used as control plots and half were fertilized with sewage sludge in a single event in 2003, applying manually to the soil surface 5 kg m-2 . The sludge came from a municipal urban wastewater treatment plant located in Tudela (Navarra, Spain), and it was sludge previously dried to 28% dry matter by centrifugation. To accurately assess vegetation changes, a 1x1 m permanent plot was placed in the centre of each macro-plot. Every year for 14 consecutive years (from 2004 to 2017), at the end of June, vegetation was sampled by R. Ibáñez, who identified and recorded every vascular plant species present in each of the permanent plots. The 1x1 m permanent plots were divided into 100 10x10 cm subplots to measure species abundance (frequency) by counting the number of 10x10 cm subplots in which the species was present (presence was recorded if shoots overlapped with the sampling unit/subplot, not according to rooted plants). Richness, synchrony and stability measures: Species richness in each permanent plot was measured both as cumulative species richness, counting the number of species found at least once in a permanent plot during the 14 years of the study, and as mean species richness, averaging the number of species found in a permanent plot over the 14 years (Lepš et al., 2018). Community-level synchrony for each permanent plot was calculated using the log variance ratio index (“Logvar”), which is the log-transformation of the ratio of observed to expected variance (i.e. the ratio of variance of the total community abundance to the sum of variances of the abundance of each species; Lepš et al., 2018; Roscher et al., 2011). Stability at both the community and the species level was calculated as the inverse of the coefficient of variation (CV-1) across years of cumulative or individual specie

1. Understanding the mechanisms underlying community stability has become an urgent need to protect ecosystems from global change and resulting biodiversity loss. While community stability can be influenced by species richness, synchrony in annual fluctuations of species, species stability and functional traits, the relative contributions of these drivers to stability are still unclear. In semi-natural grasslands, land-use changes such as fertilization might affect stability by decreasing richness and influencing year-to-year fluctuations. In addition, they can promote long-term directional trends, shifting community composition and influencing grassland maintenance. Thus, it is important to consider how species and community stability vary year-to-year but also in the long term., 2. Using a 14-year vegetation time series of a species-rich semi-natural Mediterranean grassland, we studied the relative importance of richness, synchrony, species stability and functional traits on community stability. To assess land-use change effects on stability, we applied a fertilization treatment. To distinguish stability patterns produced by year-to-year fluctuations from those caused by long-term trends, we compared the results obtained using a detrending approach from those without detrending., 3. Independently of the treatment and approach applied, the most stable communities were those composed of asynchronous species with low specific leaf area. Fertilization decreased year-to-year and long-term community stability by increasing community-weighted mean of specific leaf area, decreasing species stability or also reducing richness in the case of year-to-year stability. Additionally, traits such as seed mass had an indirect effect on stability through synchrony. Long-term trends appeared in control and fertilized plots (due to fertilization), decreasing community and species stability and leading to differences in the relationships found between community stability and some of its drivers. This reflects the importance of accounting for the effect of temporal trends on community and species stability using both a long-term and a year-to-year approach., 4. Synthesis. Stability is influenced by richness, synchrony and functional traits. Fertilization decreases species and community stability by promoting long-term trends in species composition, favouring competitive species and decreasing richness. Studying stability at the community level and species level, and accounting for the effect of trends is essential to understand stability and its drivers more comprehensively.

Therapeutic perspectives of bone tumors such as osteosarcoma remain restricted due to the inefficacy of current treatments. We propose here the construction of a novel anticancer squalene-based nanomedicine with bone affinity and retention capacity. A squalenyl-hydroxybisphosphonate molecule was synthetized by chemical conjugation of a 1-hydroxyl-1,1-bisphosphonate moiety to the squalene chain. This amphiphilic compound was inserted onto squalenoyl-gemcitabine nanoparticles using the nanoprecipitation method. The co-assembly led to nanoconstructs of 75 nm, with different morphology and colloidal properties. The presence of squalenyl-hydroxybisphosphonate enhanced the nanoparticles binding affinity for hydroxyapatite, a mineral present in the bone. Moreover, the in vitro anticancer activity was preserved when tested in commercial and patient-treated derived pediatric osteosarcoma cells. Further in vivo studies will shed light on the potential of these nanomedicines for the treatment of bone sarcomas.

Extensive pig systems are gaining importance as quality production systems and as the standard for sustainable rural development and animal welfare. However, the effects of natural foods on Salmonella epidemiology remain unknown. Herein, we assessed the presence of Salmonella and the composition of the gut microbiota in pigs from both Salmonella-free and high Salmonella prevalence farms. In addition, risk factors associated with the presence of Salmonella were investigated. The pathogen was found in 32.2% of animals and 83.3% of farms, showing large differences in prevalence between farms. Most isolates were serovars Typhimurium monophasic (79.3%) and Bovismorbificans (10.3%), and exhibited a multi-drug resistance profile (58.6%). Risk factor analysis identified feed composition, type/variety of vegetation available, and silos’ cleaning/disinfection as the main factors associated with Salmonella prevalence. Clear differences in the intestinal microbiota were found between Salmonella-positive and Salmonella-negative populations, showing the former with increasing Proteobacteria and decreasing Bacteroides populations. Butyrate and propionate producers including Clostridium, Turicibacter, Bacteroidaceae_uc, and Lactobacillus were more abundant in the Salmonella-negative group, whereas acetate producers like Sporobacter, Escherichia or Enterobacter were more abundant in the Salmonella-positive group. Overall, our results suggest that the presence of Salmonella in free-range pigs is directly related to the natural vegetation accessible, determining the composition of the intestinal microbiota.

There are important challenges when investigating individual post-translational modifications (PTMs) or protein interaction network and delineating if PTMs or their changes and cross-talks are involved during infection, disease initiation or as a result of disease progression. Proteomics and in silico approaches now offer the possibility to complement each other to further understand the regulatory involvement of these modifications in parasites and infection biology. Accordingly, the current review highlights key expressed or altered proteins and PTMs are invisible switches that turn on and off the function of most of the proteins. PTMs include phosphorylation, glycosylation, ubiquitylation, palmitoylation, myristoylation, prenylation, acetylation, methylation, and epigenetic PTMs in P. falciparum which have been recently identified. But also other low-abundant or overlooked PTMs that might be important for the parasite's survival, infectivity, antigenicity, immunomodulation and pathogenesis. We here emphasize the PTMs as regulatory pathways playing major roles in the biology, pathogenicity, metabolic pathways, survival, host-parasite interactions and the life cycle of P. falciparum. Further validations and functional characterizations of such proteins might confirm the discovery of therapeutic targets and might most likely provide valuable data for the treatment of P. falciparum, the main cause of severe malaria in human.

The mechanistic (or mammalian) target of rapamycin (mTOR) is considered as a critical regulatory enzyme involved in essential signaling pathways affecting cell growth, cell proliferation, protein translation, regulation of cellular metabolism, and cytoskeletal structure. Also, mTOR signaling has crucial roles in cell homeostasis via processes such as autophagy. Autophagy prevents many pathogen infections and is involved on immunosurveillance and pathogenesis. Immune responses and autophagy are therefore key host responses and both are linked by complex mTOR regulatory mechanisms. In recent years, the mTOR pathway has been highlighted in different diseases such as diabetes, cancer, and infectious and parasitic diseases including leishmaniasis, toxoplasmosis, and malaria. The current review underlines the implications of mTOR signals and intricate networks on pathogen infections and the modulation of this master regulator by parasites. Parasitic infections are able to induce dynamic metabolic reprogramming leading to mTOR alterations in spite of many other ways impacting this regulatory network. Accordingly, the identification of parasite effects and interactions over such a complex modulation might reveal novel information regarding the biology of the abovementioned parasites and might allow the development of therapeutic strategies against parasitic diseases. In this sense, the effects of inhibiting the mTOR pathways are also considered in this context in the light of their potential for the prevention and treatment of parasitic diseases.

Checkpoint with forkhead-associated and ring finger domains (CHFR) has been proposed as a predictive and prognosis biomarker for different tumor types, but its role in pancreatic ductal adenocarcinoma (PDAC) remains unknown. The aim of this study was two-pronged: to review the role of CHFR in PDAC and evaluating CHFR as a potential predictive biomarker in this disease. For this purpose, we first explored the CHFR messenger (m)RNA expression and promoter methylation through the TCGA database. Secondly, the CHFR expression and promoter methylation were prospectively evaluated in a cohort of patients diagnosed with borderline (n = 19) or resectable (n = 16) PDAC by immunohistochemistry (IHC), methylation specific-PCR (MSP), and pyrosequencing. The results from the TCGA database showed significant differences in terms of progression-free survival (PFS) and overall survival (OS) based on the CHFR mRNA expression, which was likely independent from the promoter methylation. Importantly, our results showed that in primarily resected patients and also the entire cohort, a higher CHFR expression as indicated by the higher IHC staining intensity might identify patients with longer disease-free survival (DFS) and OS, respectively. Similarly, in the same cohorts, patients with lower methylation levels by pyrosequencing showed significantly longer OS than patients without this pattern. Both, the CHFR expression intensity and its promoter methylation were established as independent prognostic factors for PFS and OS in the entire cohort. In contrast, no significant differences were found between different methylation patterns for CHFR and the response to taxane-based neoadjuvant treatment. These results suggest the potential role of the higher expression of CHFR and the methylation pattern of its promoter as potential prognostic biomarkers in PDAC, thus warranting further comprehensive studies to extend and confirm our preliminary findings.

A low-power ultrasonic communication front end conceived for cargo container monitoring is presented. Two piezoelectric transducers (PTs) operating at 40 kHz are fixed to the metallic wall by means of a customized magnetic case, allowing a noninvasive inside-outside communication preserving the container integrity. Once the resulting ultrasonic channel is characterized, an experimental measurement setup based on field-programmable gate array (FPGA) is implemented for testing some basic modulation and detection schemes in terms of bit error rate (BER), also considering their robustness against undesired mechanical and electromagnetic perturbations. On this basis, a compact digital differential binary phase-shift keying (DBPSK) modulator using a square carrier signal is proposed. Frequency and amplitude tracking algorithms are designed for optimizing the quality and robustness of the data transmission. Finally, a low-power low-rate (up to few kb/s) architectures based on the previous elements are presented. All the proposed contributions are experimentally validated.

Understanding the effects of landscape fragmentation on global bumblebee declines requires going beyond estimates of abundance and richness and evaluating changes in community composition and trophic and competitive interactions. We studied the effects of forest fragmentation in a Scandinavian landscape that combines temperate forests and croplands. For that, we evaluated how forest fragmentation features (patch size, isolation and shape complexity, percentage of forest in the surroundings) as well as local flowering communities influenced bumblebee abundance, richness and community composition in 24 forest patches along a fragmentation gradient. In addition, we assessed the effect of fragmentation on bumblebee–plant network specialization (H2′), and potential inter- and intraspecific competition via shared plants. Patch isolation was associated with lower bumblebee abundance, whereas flower density was positively related to both bumblebee abundance and richness. Overall, forest fragmentation reduced the abundance of forest-specialists while increasing the abundance of open-habitat species. Patches with complex shapes and few flowers showed more generalized bumblebee–plant networks (i.e., fewer specific interactions). Patch shape complexity and the percentage of forest also modified inter- and intraspecific competitive interactions, with habitat generalists outcompeting forest specialists in fragmented areas. Understanding these mechanisms is necessary to anticipate to the impact of forest fragmentation on bumblebee decline.

[Rationale]: The characterization of new genetic alterations is essential to assign effective personalized therapies in non–small cell lung cancer (NSCLC). Furthermore, finding stratification biomarkers is essential for successful personalized therapies. Molecular alterations of YES1, a member of the SRC (proto-oncogene tyrosine-protein kinase Src) family kinases (SFKs), can be found in a significant subset of patients with lung cancer., [Objectives]: To evaluate YES1 (v-YES-1 Yamaguchi sarcoma viral oncogene homolog 1) genetic alteration as a therapeutic target and predictive biomarker of response to dasatinib in NSCLC., [Methods]: Functional significance was evaluated by in vivo models of NSCLC and metastasis and patient-derived xenografts. The efficacy of pharmacological and genetic (CRISPR [clustered regularly interspaced short palindromic repeats]/Cas9 [CRISPR-associated protein 9]) YES1 abrogation was also evaluated. In vitro functional assays for signaling, survival, and invasion were also performed. The association between YES1 alterations and prognosis was evaluated in clinical samples., [Measurements and Main Results]: We demonstrated that YES1 is essential for NSCLC carcinogenesis. Furthermore, YES1 overexpression induced metastatic spread in preclinical in vivo models. YES1 genetic depletion by CRISPR/Cas9 technology significantly reduced tumor growth and metastasis. YES1 effects were mainly driven by mTOR (mammalian target of rapamycin) signaling. Interestingly, cell lines and patient-derived xenograft models with YES1 gene amplifications presented a high sensitivity to dasatinib, an SFK inhibitor, pointing out YES1 status as a stratification biomarker for dasatinib response. Moreover, high YES1 protein expression was an independent predictor for poor prognosis in patients with lung cancer., [Conclusions]: YES1 is a promising therapeutic target in lung cancer. Our results provide support for the clinical evaluation of dasatinib treatment in a selected subset of patients using YES1 status as predictive biomarker for therapy.

Selenoesters and the selenium isostere of phthalic anhydride are bioactive selenium compounds with a reported promising activity in cancer, both due to their cytotoxicity and capacity to reverse multidrug resistance. Herein we evaluate the antiviral, the biofilm inhibitory, the antibacterial and the antifungal activities of these compounds. The selenoanhydride and 7 out of the 10 selenoesters were especially potent antiviral agents in Vero cells infected with herpes simplex virus-2 (HSV-2). In addition, the tested selenium derivatives showed interesting antibiofilm activity against Staphylococcus aureus and Salmonella enterica serovar Typhimurium, as well as a moderate antifungal activity in resistant strains of Candida spp. They were inactive against anaerobes, which may indicate that the mechanism of action of these derivatives depends on the presence of oxygen. The capacity to inhibit the bacterial biofilm can be of particular interest in the treatment of nosocomial infections and in the coating of surfaces of prostheses. Finally, the potent antiviral activity observed converts these selenium derivatives into promising antiviral agents with potential medical applications.

Root performance represents a target factor conditioning plant development under drought conditions. Moreover, recent root phenotyping studies remark relevant differences on functionality of the different root types. However, despite its relevance, the performance of different types of roots such as primary/taproot (tapR) and lateral/fibrous roots (fibR) under water stress conditions is largely unknown. In the current study, the impact of water stress on target C and N metabolism (namely sucrose and proline) processes were characterized in tapR and fibR of Medicago truncatula plants exposed to different water stress severity regimes (moderate versus severe). While both root types exhibit some common responses to face water stress, the study highlighted important physiological and metabolic differences between them. The tapR proved to have an essential role on carbon and nitrogen partitioning rather than just on storage. Moreover, this root type showed a higher resilience towards water deficit stress. Sucrose metabolization at sucrose synthase level was early blocked in this tissue together with a selective accumulation of some amino acids such as proline and branched chain amino acids, which may act as alternative carbon sources under water deficit stress conditions. The decline in respiration, despite the over-accumulation of carbon compounds, suggests a modulation at sucrose cleavage level by sucrose synthase and invertase. These data not only provide new information on the carbon and nitrogen metabolism modulation upon water deficit stress but also on the different role, physiology, and metabolism of the taproot and fibrous roots. In addition, obtained results highlight the fact that both root types show distinct performance under water deficit stress; this factor can be of great relevance to improve breeding programs for increasing root efficiency under adverse conditions.

Huntington’s disease (HD) is a dominantly inherited neurodegenerative disease caused by expansion of a polyglutamine tract in the huntingtin protein. HD symptoms include severe motor, cognitive, and psychiatric impairments that result from dysfunction and later degeneration of medium-sized spiny neurons (MSNs) in the striatum. A key early pathogenic mechanism is dysregulated synaptic transmission due to enhanced surface expression of juvenile NMDA-type glutamate receptors containing GluN3A subunits, which trigger the aberrant pruning of synapses formed by cortical afferents onto MSNs. Here, we tested the therapeutic potential of silencing GluN3A expression in YAC128 mice, a well-established HD model. Recombinant adeno-associated viruses encoding a short-hairpin RNA against GluN3A (rAAV-shGluN3A) were generated, and the ability of different serotypes to transduce MSNs was compared. A single injection of rAAV9-shGluN3A into the striatum of 1-month-old mice drove potent (>90%) and long-lasting reductions of GluN3A expression in MSNs, prevented dendritic spine loss and improved motor performance in YAC128 mice. Later delivery, when spine pathology is already apparent, was also effective. Our data provide proof-of-concept for GluN3A silencing as a beneficial strategy to prevent or reverse corticostriatal disconnectivity and motor impairment in HD and support the use of RNAi-based or small-molecule approaches for harnessing this therapeutic potential.

Salmonella is a major foodborne pathogen causing important zoonosis worldwide. Pigs asymptomatically infected in mesenteric lymph nodes (MLN) can be intermittent shedders of the pathogen through faeces, being considered a major source of human infections. European baseline studies of fattening pig salmonellosis are based on Salmonella detection in MLN. This work studies the relationship between Salmonella infection in MLN and intestinal content (IC) shedding at slaughter and the relationship between the presence of the pathogen and the serologic status at farm level. Mean Salmonella prevalence in the selected pigs (vertically integrated production system of Navarra, Spain) was 7.2% in MLN, 8.4% in IC and 9.6% in serum samples. In this low–moderate prevalence context, poor concordance was found between MLN infection and shedding at slaughter and between bacteriology and serology. In fact, most of shedders were found uninfected in MLN (83%) or carrying different Salmonella strains in MLN and in IC (90%). The most prevalent Salmonellae were Typhimurium resistant to ACSSuT ± Nx or ASSuT antibiotic families, more frequently found invading the MLN (70%) than in IC (33.9%). Multivariable analysis revealed that risk factors associated with the presence of Salmonella in MLN or in IC were different, mainly related either to good hygiene practices or to water and feed control, respectively. Overall, in this prevalence context, detection of Salmonella in MLN is an unreliable predictor of faecal shedding at abattoir, indicating that subclinical infections in fattening pigs MLN could have limited relevance in the IC shedding.

The brucellae are facultative intracellular bacteria that cause a worldwide extended zoonosis. One of the pathogenicity mechanisms of these bacteria is their ability to avoid rapid recognition by innate immunity because of a reduction of the pathogen-associated molecular pattern (PAMP) of the lipopolysaccharide (LPS), free-lipids, and other envelope molecules. We investigated the Brucella homologs of lptA, lpxE, and lpxO, three genes that in some pathogens encode enzymes that mask the LPS PAMP by upsetting the core-lipid A charge/hydrophobic balance. Brucella lptA, which encodes a putative ethanolamine transferase, carries a frame-shift in B. abortus but not in other Brucella spp. and phylogenetic neighbors like the opportunistic pathogen Ochrobactrum anthropi. Consistent with the genomic evidence, a B. melitensis lptA mutant lacked lipid A-linked ethanolamine and displayed increased sensitivity to polymyxin B (a surrogate of innate immunity bactericidal peptides), while B. abortus carrying B. melitensis lptA displayed increased resistance. Brucella lpxE encodes a putative phosphatase acting on lipid A or on a free-lipid that is highly conserved in all brucellae and O. anthropi. Although we found no evidence of lipid A dephosphorylation, a B. abortus lpxE mutant showed increased polymyxin B sensitivity, suggesting the existence of a hitherto unidentified free-lipid involved in bactericidal peptide resistance. Gene lpxO putatively encoding an acyl hydroxylase carries a frame-shift in all brucellae except B. microti and is intact in O. anthropi. Free-lipid analysis revealed that lpxO corresponded to olsC, the gene coding for the ornithine lipid (OL) acyl hydroxylase active in O. anthropi and B. microti, while B. abortus carrying the olsC of O. anthropi and B. microti synthesized hydroxylated OLs. Interestingly, mutants in lptA, lpxE, or olsC were not attenuated in dendritic cells or mice. This lack of an obvious effect on virulence together with the presence of the in

[EN] This research presents the group of green vehicle routing problems with environmental costs translated into money versus production of noise, pollution and fuel consumption. This research is focused on multi-objective green logistics optimization. Optimality criteria are environmental costs: minimization of amount of money paid as externality cost for noise, pollution and costs of fuel versus minimization of noise, pollution and fuel consumption themselves. Some mixed integer programming formulations of multi-criteria vehicle routing problems have been considered. Mathematical models were formulated under assumption of existence of asymmetric distance-based costs and use of homogeneous fleet. The exact solution methods are applied for finding optimal solutions. The software used to solve these models is the CPLEX solver with AMPL programming language. The researchers were able to use real data from a Spanish company of groceries. Problems deal with green logistics for routes crossing the Spanish regions of Navarre, Basque Country and La Rioja. Analyses of obtained results could help logistics managers to lead the initiative in area of green logistics by saving money paid for environmental costs as well as direct cost of fuel and minimization of pollution and noise.

[EN] This research concerns multi-criteria vehicle routing problems. Mathematical models are formulated with mixed-integer programming. We consider maximization of capacity of truck vs. minimization of utilization of fuel, carbon emission and production of noise. The problems deal with green logistics for routes crossing the Western Pyrenees in Navarre, Basque Country and La Rioja, Spain. We consider heterogeneous fleet of trucks. Different types of trucks have not only different capacities, but also require different amounts of fuel for operations. Consequently, the amount of carbon emission and noise vary as well. Companies planning delivery routes must consider the trade-off between the financial and environmental aspects of transportation. Efficiency of delivery routes is impacted by truck size and the possibility of dividing long delivery routes into smaller ones. The results of computational experiments modeled after real data from a Spanish food distribution company are reported. Computational results based on formulated optimization models show some balance between fleet size, truck types, utilization of fuel, carbon emission and production of noise. As a result, the company could consider a mixture of trucks sizes and divided routes for smaller trucks. Analyses of obtained results could help logistics managers lead the initiative in environmental conservation by saving fuel and consequently minimizing pollution.

[EN] Since its appearance in the 1990s, horizontal collaboration (HC) practices have revealed themselves as catalyzers for optimizing the distribution of goods in freight transport logistics. After introducing the main concepts related to HC, this paper offers a literature review on the topic and provides a classification of best practices in HC. Then, the paper analyses the main benefits and optimization challenges associated with the use of HC at the strategic, tactical, and operational levels. Emerging trends such as the concept of ` green' or environmentally- friendly HC in freight transport logistics are also introduced. Finally, the paper discusses the need of using hybrid optimization methods, such as simheuristics and learnheuristics, in solving some of the previously identified challenges in real- life scenarios dominated by uncertainty and dynamic conditions.

This study was intended to investigate the gut microbiota composition in fattening pigs reared under extensive conditions and clustered by high prevalence or absence of Salmonella spp in faeces, at herd level

Although responsiveness of N2-fixing plants to elevated CO2 conditions have been analyzed in previous studies, important uncertainties remain in relation to the effect enhanced CO2 in nodule proteomic profile and its implication in leaf responsiveness. The aim of our study was to deepen our understanding of the relationship between leaf and nodule metabolism of N2-fixing alfalfa plants after long-term exposure to elevated CO2. After 30-day exposure to elevated CO2, plants showed photosynthetic down-regulation with reductions in the light-saturated rate of CO2 assimilation (Asat) and the maximum rate of rubisco carboxylation (Vcmax). Under elevated CO2 conditions, the rubisco availability limited potential photosynthesis by around 12 %, which represented the majority of the observed fall in Vcmax. Photosynthetic down-regulation has been associated with decreased N availability even if those plants are capable to assimilate N2. Diminishment in shoot N demand (as reflected by the lower rubisco and leaf N content) suggests that the lower aboveground N requirements affected negatively nodule performance. In this condition, specific nodule activity was reduced due to an effect on nodule metabolism that manifested as a lower amount of nitrogenase reductase. Moreover, the nodule proteomic approach also revealed that nodule functioning was altered simultaneously in various enzyme quantity apart from nitrogenase. At elevated CO2, the tricarboxylic acid cycle was also altered with a reduced amount of isocitrate synthase protein. The nodule proteome analysis also revealed the relaxation of the antioxidant system as shown by a decline in the amount of catalase and isoflavone reductase protein.

The aim of the present study was to analyse the effect of elevated CO 2 at different temperature, inoculation with three Sinorhizobium meliloti strains and growth in two seasons (summer and autumn) on plant growth, photosynthesis and N 2 fixation (estimated as apparent nitrogenase activity, ANA). One of the objectives was to demonstrate that most efficient N 2-fixing strains could improve alfalfa forage production. The second objective was to test if S. meliloti strains alter their efficiency under climate change conditions. The third objective was to show if seasonal changes may modify alfalfa productivity. Interaction of CO 2 and temperature enhanced alfalfa dry matter in both seasons; however, plants produced more dry matter in autumn than in summer, due to the negative effect of elevated summer temperature on N 2-fixation. Higher yield in summer corresponded to plants in symbiosis with 102F78 strain being not related to enhanced nodule dry matter or apparent nitrogenase activity but to putative lower carbon consumption for N 2 fixing process. Contrariwise, in autumn the highest yield was obtained by 102F34 as a consequence of increased nodule dry matter induced under elevated CO 2 and temperature. © 2011 Elsevier B.V.

Elevated CO 2 may decrease alfalfa forage quality and in vitro digestibility through a drop in crude protein and an enhancement of fibre content. The aim of the present study was to analyse the effect of elevated CO 2, elevated temperature and Sinorhizobium meliloti strains (102F78, 102F34 and 1032 GMI) on alfalfa yield, forage quality and in vitro dry matter digestibility. This objective is in line with the selection of S. meliloti strains in order to maintain high forage yield and quality under future climate conditions. Plants inoculated with the 102F34 strain showed more DM production than those inoculated with 1032GMI; however, these strains did not show significant differences with 102F78 plants. Neutral or acid detergent fibres were not enhanced in plants inoculated with the 102F34 strain under elevated CO 2 or temperature and hence, in vitro dry matter digestibility was unaffected. Crude protein content, an indicator of forage quality, was negatively related to shoot yield. Plants inoculated with 102F78 showed a similar shoot yield to those inoculated with 102F34, but had higher crude protein content at elevated CO 2 and temperature. Under these climate change conditions, 102F78 inoculated plants produced higher quality forage. However, the higher digestibility of plants inoculated with the 102F34 strain under any CO 2 or temperature conditions makes them more suitable for growing under climate change conditions. In general, elevated CO 2 in combination with high temperature (Climate Change scenario) reduced IVDMD and CP content and enhanced fibre content, which means that animal production will be negatively affected. © 2012 Elsevier GmbH.

Future climatic conditions, including rising atmospheric CO2 and temperature may increase photosynthesis and, consequently, plant production. A larger knowledge of legume performance under the predicted growth conditions will be crucial for safeguarding crop management and extending the area under cultivation with these plants in the near future. N2 fixation is a key process conditioning plant responsiveness to varying growth conditions. Moreover, it is likely to increase under future environments, due to the higher photosynthate availability, as a consequence of the higher growth rate under elevated CO2. However, as described in the literature, photosynthesis performance is frequently down-regulated (acclimated) under long-term exposure to CO2, especially when affected by stressful temperature and water availability conditions. As growth responses to elevated CO2 are dependent on sink-source status, it is generally accepted that down-regulation occurs in situations with insufficient plant C sink capacity. Alfalfa management involves the cutting of shoots, which alters the source-sink relationship and thus the photosynthetic behaviour. As the growth rate decreases at the end of the pre-cut vegetative growth period, nodulated alfalfa plants show photosynthetic down-regulation, but during regrowth following defoliation, acclimation to elevated CO2 disappears. The shoot harvest also leads to a drop in mineral N uptake and C translocation to the roots, resulting in a reduction in N2 fixation due to the dependence on photosynthate supply to support nodule function. Therefore, the production of new shoots during the first days following cutting requires the utilization of reduced C and N compounds that have been stored previously in reserve organs. The stored reserves are mediated by phytohormones such as methyl jasmonate and abscisic acid and in situations where water stress reduces shoot production this potentially enables the enhancement of taproot protein levels in no

Edelfosine is the prototype molecule of a family of anticancer drugs collectively known as synthetic alkyl-lysophospholipids. This drug holds promise as a selective antitumor agent, and a number of preclinical assays are in progress. In this study, we observe the accumulation of edelfosine in brain tissue after its oral administration in Compritol (R) and Precirol (R) lipid nanoparticles (LN). The high accumulation of edelfosine in brain was due to the inhibition of P-glycoprotein by Tween (R) 80, as verified using a P-glycoprotein drug interaction assay. Moreover, these LN were tested in vitro against the C6 glioma cell line, which was later employed to establish an in vivo xenograft mouse model of glioma. In vitro studies revealed that edelfosine-loaded LN induced an antiproliferative effect in C6 glioma cell line. In addition, in vivo oral administration of drug-loaded LN in NMRI nude mice bearing a C6 glioma xenograft tumor induced a highly significant reduction in tumor growth (p<0.01) 14 days after the beginning of the treatment. Our results showed that Tween (R) 80 coated Compritol (R) and Precirol (R) LN can effectively inhibit the growth of C6 glioma cells in vitro and suggest that edelfosine-loaded LN represent an attractive option for the enhancement of antitumor activity on brain tumors in vivo.

[Purpose]: Mantle cell lymphoma (MCL) and chronic lymphocytic leukemia (CLL) remain B-cell malignancies with limited therapeutic options. The present study investigates the in vitro and in vivo effect of the phospholipid ether edelfosine (1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine) in MCL and CLL. [Experimental Design]: Several cell lines, patient-derived tumor cells, and xenografts in severe combined immunodeficient mice were used to examine the anti-MCL and anti-CLL activity of edelfosine. Furthermore, we analyzed the mechanism of action and drug biodistribution of edelfosine in MCL and CLL tumor-bearing severe combined immunodeficient mice. [Results]: Here, we have found that the phospholipid ether edelfosine was the most potent alkyl-lysophospholipid analogue in killing MCL and CLL cells, including patient-derived primary cells, while sparing normal resting lymphocytes. Alkyl-lysophospholipid analogues ranked edelfosine > perifosine ≫ erucylphosphocholine ≥ miltefosine in their capacity to elicit apoptosis in MCL and CLL cells. Edelfosine induced coclustering of Fas/CD95 death receptor and rafts in MCL and CLL cells. Edelfosine was taken up by malignant cells, whereas normal resting lymphocytes hardly incorporated the drug. Raft disruption by cholesterol depletion inhibited drug uptake, Fas/CD95 clustering, and edelfosine-induced apoptosis. Edelfosine oral administration showed a potent in vivo anticancer activity in MCL and CLL xenograft mouse models, and the drug accumulated dramatically and preferentially in the tumor. [Conclusions]: Our data indicate that edelfosine accumulates and kills MCL and CLL cells in a rather selective way, and set coclustering of Fas/CD95 and lipid rafts as a new framework in MCL and CLL therapy. Our data support a selective antitumor action of edelfosine.